阅读说明：本技术 使用ccr3-抑制剂治疗衰老相关损伤的方法及组合物 (Methods and compositions for treating aging-related injuries using CCR 3-inhibitors ) 是由 史蒂文·P·布雷斯韦特 S·樱井·南 卡罗伊·尼科利希 桑凯特·V·雷杰 阿诺德·E·J·泰 于 2019-09-25 设计创作，主要内容包括：提供使用CCR3调节剂来改善神经退行性疾病的方法。这些方法包括向对象施用治疗有效量的CCR3调节剂,其中在认知、运动或其他受神经退行影响的功能上会有伴随性的改善。本发明方法可改善认知的认知性及运动疾病包括阿兹海默氏症、帕金森氏病、额颞叶型痴呆、亨丁顿氏症、肌萎缩性侧索硬化症、多发性硬化症、青光眼、肌强直性营养不良、血管性痴呆、进行性核上性麻痹症。(Methods of using modulators of CCR3 to ameliorate neurodegenerative diseases are provided. These methods comprise administering to a subject a therapeutically effective amount of a CCR3 modulator wherein there is a concomitant improvement in cognition, exercise or other neurodegenerative affected functions. Cognitive and motor disorders that can improve cognition in the methods of the invention include Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, vascular dementia, progressive supranuclear palsy.)

1. A method of treating a neurodegenerative disease in a subject diagnosed with the neurodegenerative disease, the method comprising administering a therapeutically effective amount of a compound of formula 1 to treat the neurodegenerative disease in the subject:

wherein

A is CH₂O or N-C_1-6-an alkyl group;

R¹is selected from

·NHR^1.1、NMeR^1.1；

·NHR^1.2、NMeR^1.2；

·NHCH₂-R^1.3；

·NH-C_3-6-cycloalkyl in which optionally one carbon atom is replaced by a nitrogen atom, wherein the ring is optionally substituted by a group selected from C_1-6Alkyl, O-C_1-6Alkyl, NHSO₂Phenyl, NHCONH-phenyl, halogen, CN, SO₂-C_1-6-alkyl, COO-C_1-6-one or two residue substitutions of alkyl;

·C_{9 or 10}-bicyclic ring, wherein one or two carbon atoms are replaced by nitrogen atoms, the ring system being bonded to the basic structure of formula 1 via nitrogen atoms, wherein the ring system is optionally selected from C_1-6-alkyl, COO-C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, NO₂Halogen, CN, NHSO ₂-C_1-6-alkyl, methoxy-phenyl substituted with one or two residues;

selected from NHCH (pyridyl) CH₂COO-C_1-6Alkyl, NHCH (CH)₂O-C_1-6-alkyl) -benzimidazolyl, optionally substituted with halogen or CN; or

1-aminocyclopentyl radical, which is optionally substituted by methyl-Oxadiazole substitution;

R^1.1is phenyl, optionally via a radical selected from C_1-6Alkyl radical, C_2-6-alkenyl, C_2-6-alkynyl, C_1-6-haloalkyl group, C_1-6alkylene-OH, C_2-6-alkenylene-OH, C_2-6-alkynylene-OH, CH₂CON(C_1-6-alkyl groups)₂、CH₂NHCONH-C_3-6-cycloalkyl, CN, CO-pyridyl, CONR^1.1.1R^1.1.2、COO-C_1-6Alkyl, N (SO)₂-C_1-6-alkyl) (CH₂CON(C_1-4-alkyl groups)₂)O-C_1-6-alkyl, O-pyridyl, SO₂-C_1-6Alkyl, SO₂-C_1-6alkylene-OH, SO₂-C_3-6-cycloalkyl, SO₂Piperidinyl, SO₂NH-C_1-6Alkyl, SO₂N(C_1-6-alkyl groups)₂Halogen, CN, CO-morpholinyl, CH₂-one or two residues of pyridyl, or optionally substituted by a group selected from C_1-6-alkyl, NHC_1-6-alkyl and a heterocycle substituted with one or two residues of ═ O;

R^1.1.1is H, C_1-6Alkyl radical, C_3-6-cycloalkyl, C_1-6-haloalkyl, CH₂CON(C_1-6-alkyl groups)₂、CH₂CO-Azacyclobutyl, C_1-6alkylene-C_3-6-cycloalkyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, C_1-6alkylene-OH or thiadiazolyl, optionally via C_1-6-alkyl substitution;

R^1.1.2is H, C_1-6Alkyl, SO₂C_1-6-an alkyl group;

or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one N or O replacing a carbon atom on the ring, and optionally via a substituent selected from C _1-6Alkyl radical, C_1-4-alkylene-OH, ═ O, substituted with one or two residues;

or

R^1.1Is phenyl, in which two adjacent residues together form a five-or six-membered carbocyclic aromatic or nonaromatic ring which optionally independently of one another contains a substitution on the ringOne or two of carbon atoms N, S or SO₂Wherein the five-or six-membered carbocyclic aromatic or nonaromatic ring is optionally via C_1-4-alkyl or ═ O substitution;

R^1.2is selected from

Heteroaryl, optionally via a radical selected from C_1-6Alkyl radical, C_2-6-alkenyl, C_2-6-alkynyl, C_3-6-cycloalkyl, CH₂COO-C_1-6Alkyl, CONR^1.2.1R^1.2.2、COR^1.2.3、COO-C_1-6Alkyl, CONH₂、O-C_1-6Alkyl, halogen, CN, SO₂N(C_1-6-alkyl groups)₂Is substituted by one or two residues, or heteroaryl, optionally substituted by a group selected from C_1-6-one or two residue substitutions of alkyl;

heteroaryl, optionally substituted by a five-or six-membered carbocyclic non-aromatic ring containing, independently of one another, two N, O, S or SO replacing a carbon atom on the ring₂；

Aromatic or non-aromatic C_{9 or 10}Bicyclic ring, in which one or two carbon atoms are replaced by N, O or S, said aromatic or non-aromatic C_{9 or 10}-each of the bicyclic rings is optionally selected from N (C)_1-6-alkyl groups)₂、CONH-C_1-6-alkyl, ═ O substituted with one or two residues;

a heterocyclic non-aromatic ring, optionally substituted with pyridyl;

4, 5-dihydro-naphtho [2,1-d ]]Thiazoles, optionally via NHCO-C_1-6-an alkyl substitution,

R^1.2.1is H, C_1-6Alkyl radical, C_1-6alkylene-C_3-6-cycloalkyl, C_1-4Alkylene-phenyl, C_1-4Alkylene-furyl radical, C_3-6-cycloalkyl, C_1-4alkylene-O-C_1-4Alkyl radical, C_1-6-haloalkyl or a five-or six-membered carbocyclic non-aromatic ring optionally containing, independently of each other, one or two N, O, S or SO replacing carbon atoms on the ring₂And optionally substituted with 4-cyclopropylmethyl-piperazinyl;

R^1.2.2is H, C_1-6-an alkyl group;

R^1.2.3is a five-or six-membered carbocyclic non-aromatic ring optionally containing, independently of one another, one or two N, O, S or SO replacing a carbon atom on the ring₂；

R^1.3Selected from phenyl, heteroaryl or indolyl, each of which is optionally selected from C_1-6Alkyl radical, C_3-6-cycloalkyl, O-C_1-6Alkyl, O-C_1-6-one or two residue substitutions of haloalkyl, phenyl, heteroaryl;

R²is selected from C_1-6Alkylene-phenyl, C_1-6-alkylene-naphthyl, and C_1-6-alkylene-heteroaryl; each of which is optionally selected from C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, O-C_1-6-one, two or three residue substitutions of haloalkyl, halogen;

R³is H, C_1-6-an alkyl group;

R⁴is H, C_1-6-an alkyl group;

or R³And R⁴Together form CH ₂-CH₂A group.

2. The method of claim 1, wherein the compound of formula 1,

a is CH₂O or N-C_1-4-an alkyl group;

R¹is selected from

·NHR^1.1、NMeR^1.1；

·NHR^1.2、NMeR^1.2；

·NHCH₂-R^1.3；

·NH-C_3-6Cycloalkyl in which optionally one carbon atom is replaced by a nitrogen atom, in which the ring is optionally substituted by a group selected from C_1-6Alkyl, O-C_1-6Alkyl, NHSO₂Phenyl, NHCONH-phenyl, halogen, CN, SO₂-C_1-6-alkyl, COO-C_1-6-one or two residue substitutions of alkyl;

·C_{9 or 10}-bicyclic ring, wherein one or two carbon atoms are replaced by nitrogen atoms and the ring system is bonded to the basic structure of formula 1 via nitrogen atoms, wherein the ring system is optionally selected from C_1-6-alkyl, COO-C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, NO₂Halogen, CN, NHSO₂-C_1-6-alkyl, m-methoxyphenyl, substituted by one or two residues;

selected from NHCH (pyridyl) CH₂COO-C_1-6Alkyl, NHCH (CH)₂O-C_1-6-alkyl) -benzimidazolyl, optionally substituted with Cl; or

1-aminocyclopentyl, optionally methyl-Substituted by diazole group;

R^1.1is phenyl, optionally via a radical selected from C_1-6Alkyl radical, C_1-6-haloalkyl, CH₂CON(C_1-6-alkyl groups)₂、CH₂NHCONH-C_3-6-cycloalkyl, CN, CONR^1.1.1R^1.1.2、COO-C_1-6Alkyl, O-C_1-6Alkyl, SO₂-C_1-6Alkyl, SO₂-C_1-6alkylene-OH, SO₂-C_3-6-cycloalkyl, SO₂Piperidinyl, SO₂NH-C_1-6Alkyl, SO₂N(C_1-6-alkyl groups)₂Halogen, CN, CO-morpholinyl, CH ₂-substitution of one or two residues of pyridyl, or R^1.1Is optionally selected from C_1-6-alkyl, NHC_1-6-alkyl, ═ O, and heterocycles substituted with one or two residues;

R^1.1.1is H, C_1-6Alkyl radical, C_3-6-cycloalkyl, C_1-6-haloalkyl, CH₂CON(C_1-6-alkyl groups)₂、CH₂CO-Azacyclobutyl, C_1-6alkylene-C_3-6-cycloalkyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, C_1-6alkylene-OH or thiadiAzolyl, optionally via C_1-6-alkyl substitution;

R^1.1.2is H, C_1-6Alkyl, SO₂C_1-6-an alkyl group;

or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one O replacing a carbon atom on the ring, and optionally via a substituent selected from CH₂One or two residue substitutions of OH;

R^1.2is selected from

Heteroaryl, optionally via a radical selected from C_1-6Alkyl radical, C_3-6-cycloalkyl, CH₂COO-C_1-6Alkyl, CONR^1.2.1R^1.2.2、COO-C_1-6Alkyl, CONH₂、O-C_1-6-alkyl, halogen, CN, CO-pyrrolidinyl, CO-morpholinyl, substituted with one or two residues, or heteroaryl, optionally substituted with a group selected from C_1-6-one or two residue substitutions of alkyl;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each of which is optionally substituted with a group selected from N (C)_1-6-alkyl groups)₂、CONH-C_1-6-alkyl, ═ O substituted with one or two residues;

piperidinyl, optionally substituted with pyridinyl;

4, 5-dihydro-naphtho [2,1-d ] ]Thiazoles, optionally via NHCO-C_1-6-alkyl substitution;

R^1.2.1is H, C_1-6-an alkyl group;

R^1.2.2is H, C_1-6-an alkyl group;

R^1.3selected from phenyl, pyrazolyl, isoAzolyl, pyrimidinyl, indolyl orOxadiazolyl, each of which is optionally selected from C_1-6Alkyl radical, C_3-6-cycloalkyl, O-C_1-6Alkyl, O-C_1-6-mono or of haloalkylTwo residue substitutions;

R²is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, O-C_1-6-haloalkyl, halogen substituted with one or two residues; or R²Is selected from CH₂-thiophenyl, optionally substituted with one or two residues selected from halogen;

R³is H, C_1-4-an alkyl group;

R⁴is H, C_1-4-an alkyl group;

or R³And R⁴Together form CH₂-CH₂A group.

3. The method of claim, wherein the compound of formula 1 is

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

·NHR^1.2、NMeR^1.2；

·NHCH₂-R^1.3；

NH-cyclohexyl, optionally via a radical selected from C_1-4Alkyl, NHSO₂-phenyl, NHCONH-phenyl, halo substituted with one or two residues;

NH-pyrrolidinyl, optionally via a radical selected from SO₂-C_1-4-alkyl, COO-C_1-4-one or two residue substitutions of alkyl;

piperidinyl, optionally via a radical selected from NHSO₂-C_1-4-alkyl, m-methoxyphenyl, substituted by one or two residues;

dihydro-indolyl, dihydro-isoindolyl, tetrahydro-quinolyl or tetrahydro-isoquinolyl, optionally via a substituent selected from C _1-4-alkyl, COO-C_1-4Alkyl radical, C_1-4Haloalkyl, O-C_1-4Alkyl, NO₂One or two residue substitutions of halogen;

selected from NHCH (pyridyl) CH₂COO-C_1-4-alkyl radical、NHCH(CH₂O-C_1-4-alkyl) -benzimidazolyl, optionally substituted with Cl; or

1-aminocyclopentyl, optionally methyl-Substituted by diazole group;

R^1.1is phenyl, optionally via a radical selected from C_1-4Alkyl radical, C_1-4-haloalkyl, CH₂CON(C_1-4-alkyl groups)₂、CH₂NHCONH-C_3-6-cycloalkyl, CN, CONR^1.1.1R^1.1.2、COO-C_1-4Alkyl, O-C_1-4Alkyl, SO₂-C_1-4Alkyl, SO₂-C_1-4alkylene-OH, SO₂-C_3-6-cycloalkyl, SO₂Piperidinyl, SO₂NH-C_1-4Alkyl, SO₂N(C_1-4-alkyl groups)₂Halogen, CO-morpholinyl, CH₂-substitution of one or two residues of pyridyl, or R^1.1Is imidazolidinyl, piperidyl, oxazacyclohexanyl, pyrazolyl, triazolyl, tetrazolyl, triazolyl, and oxaziridinyl,Azolyl group,Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each optionally selected from C_1-4-alkyl, NHC_1-4-alkyl, ═ O substituted with one or two residues;

R^1.1.1is H, C_1-6Alkyl radical, C_3-6-cycloalkyl, C_1-4-haloalkyl, CH₂CON(C_1-4-alkyl groups)₂、CH₂CO-Azacyclobutyl, C_1-4alkylene-C_3-6-cycloalkyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, C_1-4alkylene-OH or thiadiazolyl, optionally via C_1-4-alkyl substitution;

R^1.1.2is H, C_1-4Alkyl, SO₂C_1-4-an alkyl group;

Or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one O replacing a carbon atom on the ring, and optionally via a substituent selected from CH₂One or two residue substitutions of OH;

R^1.2is selected from

Pyridinyl, pyridazinyl, pyrrolyl, pyrazolyl, isoAzolyl, thiazolyl, thiadiazolyl, optionally selected from C_1-4Alkyl radical, C_3-6-cycloalkyl, CH₂COO-C_1-4Alkyl, CONR^1.2.1R^1.2.2、COO-C_1-4Alkyl, CONH₂、O-C_1-4-alkyl, halogen, CO-pyrrolidinyl, CO-morpholinyl, or R^1.2Selected from pyrazolyl, triazolyl, tetrazolyl, iso-pyrazolylAzolyl group,Oxadiazolyl, each of which is optionally selected from C_1-4-one or two residue substitutions of alkyl;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each of which is optionally substituted with a group selected from N (C)_1-4-alkyl groups)₂、CONH-C_1-4-alkyl, ═ O substituted with one or two residues;

piperidinyl, optionally substituted with pyridinyl;

4, 5-dihydro-naphtho [2,1-d ]]Thiazoles, optionally via NHCO-C_1-4-alkyl substitution;

R^1.2.1is H, C_1-4-an alkyl group;

R^1.2.2is H, C_1-4-an alkyl group;

R^1.3selected from phenyl, pyrazolyl, isoAzolyl, pyrimidinyl, indolyl orOxadiazolyl, each optionally via C_1-4Alkyl radical, C_3-6-cycloalkyl, O-C_1-4Alkyl, O-C_1-4-one or two residue substitutions of haloalkyl;

R²Is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via C_1-4Alkyl radical, C_1-4Haloalkyl, O-C_1-4-haloalkyl, halogen substituted with one or two residues; or R²Is selected from CH₂-thiophenyl, optionally substituted with one or two residues selected from halogen;

R³is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

4. The method of claim 1, wherein formula 1 is

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

·NHR^1.2、NMeR^1.2；

·NHCH₂-R^1.3；

NH-piperidinyl, optionally substituted with pyridinyl;

NH-cyclohexyl, optionally via a gas selected from t-Bu, NHSO₂-phenyl, NHCONH-phenyl, substitution of one or two residues of F;

NH-pyrrolidinyl, optionally via a radical selected from SO₂One or two residues of Me and COO-t-Bu;

piperidinyl, optionally via a radical selected from NHSO₂One or two residues of-n-Bu, m-methoxyphenylSubstitution;

dihydro-indolyl, dihydro-isoindolyl, tetrahydro-quinolyl or tetrahydro-isoquinolyl, optionally substituted by a substituent selected from Me, COOMe, CF₃、OMe、NO₂F, Br;

selected from NHCH (pyridyl) CH₂COOMe、NHCH(CH₂OMe) -benzimidazolyl, optionally substituted with Cl; or

1-aminocyclopentyl, optionally methyl-Substituted by diazole group;

R^1.1is phenyl, optionally via a catalyst selected from Me, Et, t-Bu, CF₃、CH₂CONMe₂、CH₂NHCONH-cyclohexyl, CN, CONR ^1.1.1R^1.1.2、COOMe、COOEt、OMe、SO₂Me、SO₂CH₂CH₂OH、SO₂Et、SO₂-cyclopropyl, SO₂Piperidinyl, SO₂NHEt、SO₂NMeEt, F, Cl, CO-morpholinyl, CH₂-substitution of one or two residues of pyridyl, or R^1.1Is imidazolidinyl, piperidyl, oxazacyclohexanyl, pyrazolyl, triazolyl, tetrazolyl, triazolyl, and oxaziridinyl,Azolyl group,Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each of which is optionally substituted with one or two residues selected from Me, NHMe, ═ O;

R^1.1.1is H, Me, Et, t-Bu, i-Pr, cyclopropyl, CH₂-i-Pr、CH₂-t-Bu、CH(CH₃)CH₂CH₃、CH₂CHF₂、CH₂CONMe₂、CH₂CO-AZACYCLOBUTYL, CH₂-cyclopropyl, CH₂-cyclobutyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, CH₂CH₂OH or thiadiazolyl, optionally substituted with Me;

R^1.1.2is H, Me, Et, SO₂Me、SO₂Et；

Or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one O replacing a carbon atom on the ring, and optionally via a substituent selected from CH₂One or two residue substitutions of OH;

R^1.2is selected from

Pyridinyl, pyrrolyl, pyrazolyl, isoOxazolyl, thiazolyl, thiadiazolyl, optionally substituted by a group selected from Me, Et, Pr, Bu, cyclopropyl, CH₂COOEt、CONR^1.2.1R^1.2.2、COOMe、COOEt、CONH₂One or two residues of OMe, Cl, Br, CO-pyrrolidinyl, CO-morpholinyl, or R^1.2Selected from pyrazolyl, triazolyl, tetrazolyl, iso-pyrazolylAzolyl group,(ii) oxadiazolyl, each of which is optionally substituted with Me;

Benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each of which is optionally substituted with NMe₂One or two residue substitutions of CONHMe, ═ O;

4, 5-dihydro-naphtho [2,1-d ] thiazole, optionally substituted with NHCOMe,

R^1.2.1h, Me;

R^1.2.2h, Me;

R^1.3selected from phenyl, pyrazolyl, isoAzolyl, pyrimidinyl, indolyl orOxadiazolyl, each of which is optionally substituted with one or more substituents selected from the group consisting of Me, Et, Pr, cyclopentyl, OMe, OCHF₂One or two residue substitutions;

R²is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via CH₃、CF₃、OCF₃One or two residues of F, Cl, Br, Et; or R²Is selected from CH₂-thiophenyl, optionally substituted with one or two residues selected from Cl, Br;

R³is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

5. The method of claim 1, wherein formula 1 is

A is CH₂O or NMe;

R¹is selected from

NHR^1.1，

NHR^1.2，

R^1.1Is phenyl, optionally via a catalyst selected from Me, Et, Bu, CF₃、CH₂CONMe₂、CH₂NHCONH-cyclohexyl, CN, CONR^1.1.1R^1.1.2、COOMe、COOEt、OMe、SO₂Me、SO₂CH₂CH₂OH、SO₂Et、SO₂-cyclopropyl, SO₂Piperidinyl, SO₂NHEt、SO₂NMeEt, F, Cl, CO-morpholinyl, CH₂-substitution of one or two residues of pyridyl, or R^1.1Is imidazolidinyl, piperidyl, oxazacyclohexanyl, pyrazolyl, triazolyl, tetrazolyl, triazolyl, and oxaziridinyl,Azolyl group, Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each of which is optionally substituted with one or two residues selected from Me, NHMe, ═ O;

R^1.1.1is H, Me, Et, t-Bu, i-Pr, cyclopropyl, CH₂-i-Pr、CH₂-t-Bu、CH(CH₃)CH₂CH₃、CH₂CHF₂、CH₂CONMe₂、CH₂CO-AZACYCLOBUTYL, CH₂-cyclopropyl, CH₂-cyclobutyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, CH₂CH₂OH or thiadiazolyl, optionally substituted with Me;

R^1.1.2is H, Me, Et, SO₂Me、SO₂Et；

Or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one O replacing a carbon atom on the ring, and optionally via a substituent selected from CH₂One or two residue substitutions of OH;

R^1.2is selected from

Pyridinyl, pyrrolyl, pyrazolyl, isoOxazolyl, thiazolyl, thiadiazolyl, optionally substituted by a group selected from Me, Et, Pr, Bu, cyclopropyl, CH₂COOEt、CONR^1.2.1R^1.2.2、COOMe、COOEt、CONH₂One or two residues of OMe, Cl, Br, CO-pyrrolidinyl, CO-morpholinyl, or R^1.2Selected from pyrazolyl, triazolyl, tetrazolyl, iso-pyrazolylAzolyl group,(ii) oxadiazolyl, each of which is optionally substituted with Me;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolyl, eachIs selected from NMe₂One or two residue substitutions of CONHMe, ═ O;

4, 5-dihydro-naphtho [2,1-d ] thiazole, optionally substituted with NHCOMe;

R^1.2.1H, Me;

R^1.2.2h, Me;

R²is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via CH₃、CF₃、OCF₃One or two residues of F, Cl, Br, Et;

R³is H;

R⁴is H.

6. The method of claim 1, wherein formula 1 is

A is CH₂O or NMe;

R¹is selected from

R²Is selected from

R³Is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

7. The method of claim 1, wherein the compound is a co-crystal of the formula

Wherein

R¹Is C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6-haloalkyl, halogen;

m is 1, 2 or 3;

R^2aand R^2bEach independently selected from H, C_1-6Alkyl radical, C_1-6-alkenyl, C_1-6-alkynyl, C_3-6-cycloalkyl, COO-C_1-6Alkyl, O-C_1-6Alkyl, CONR^2b.1R^2b.2A halogen;

R^2b.1is H, C_1-6Alkyl radical, C_0-4-alkyl-C_3-6-cycloalkyl, C_1-6-a haloalkyl group;

R^2b.2is H, C_1-6-an alkyl group;

or R^2b.1And R^2b.2Together are C_3-6-an alkylene group forming a heterocyclic ring with a nitrogen atom, wherein optionally one carbon atom of said heterocyclic ring is replaced by an oxygen atom;

R³is H, C_1-6-an alkyl group;

x is an anion selected from chloride, bromide, iodide, sulfate, phosphate, methanesulfonate, nitrate, maleate, acetate, benzoate, citrate, salicylate, fumarate, tartrate, dibenzoyltartrate, oxalate, succinate, benzoate, and p-toluenesulfonate;

j is 0, 0.5, 1, 1.5 or 2;

wherein the co-crystal former is selected from the group consisting of: orotic acid, hippuric acid, L-pyroglutamic acid, D-pyroglutamic acid, nicotinic acid, L- (+) -ascorbic acid, saccharin, piperazine, 3-hydroxy-2-naphthoic acid, mucic acid (galactaric acid), pamoic acid (enbo acid), stearic acid, cholic acid, deoxycholic acid, nicotinamide, isonicotinamide, succinamide, uracil, L-lysine, L-proline, D-valine, L-arginine, glycine.

8. The method of claim 1, wherein the compound is a crystalline salt of the formula,

9. the method of claim 1, wherein the compound comprises a co-crystal of at least one compound of formula (lb) according to claim 7 and a pharmaceutically acceptable carrier.

10. The method of claim 1, wherein the compound of formula 1 is administered as individual optical isomers, mixtures of individual enantiomers, racemates or as enantiomerically pure compounds.

11. The method of claim 1, wherein the compound is a pharmaceutical composition comprising one or more than one compound of the formula as an active ingredient, a first diluent, a second diluent, a binder, a disintegrant, and a lubricant,

Wherein

R¹Is H, C_1-6Alkyl radical, C_0-4-alkyl-C_3-6-cycloalkyl, C_1-6-a haloalkyl group;

R²is H, C_1-6-an alkyl group;

x is an anion selected from chloride or 1/2 dibenzoyl tartrate;

j is 1 or 2.

12. The method of claim 11, wherein the pharmaceutical composition further comprises an additional disintegrant.

13. The method of claim 11 or 12, wherein the pharmaceutical composition further comprises an additional glidant.

14. The method of claim 11, 12 or 13, wherein the diluent of the pharmaceutical composition further comprises cellulose powder, anhydrous dibasic calcium phosphate, dehydrated dibasic calcium phosphate, erythritol, low substituted hydroxypropyl cellulose, mannitol, pregelatinized starch or xylitol.

15. The method of any one of the preceding claims, wherein the neurodegenerative disease is selected from alzheimer's disease, parkinson's disease, frontotemporal dementia, huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, vascular dementia, progressive supranuclear palsy.

Brief description of the drawings

Fig. 1A depicts the sum of Double Cortin (DCX) positive cells in hippocampus of the following groups: IgG-treated C57B1/6 mice 2 and 18 months old (both groups n-18); 18-month old mice treated with compound 1 at high (n ═ 16) or low (n ═ 31) doses for 2 weeks; and 18-month old mice treated with compound 1 ("Cmpd 1") at low dose for 4 weeks (n ═ 16). Data shown are mean ± s.e.m., where P <0.05 and P < 0.001.

FIG. 1B depicts the sum of 5-bromo-2' -deoxyuridine (BrdU) positive cells in hippocampus of the following groups: IgG-treated C57B1/6 mice 2 and 18 months old (n ═ 19 in both groups); 18-month-old mice were treated with compound 1 at either high (n-16) or low (n-24) doses for 2 weeks. Data shown are mean ± s.e.m., where P < 0.001.

Figure 2A depicts the effect of compound 1 on the time spent in the novel (N) arm versus the familiar/old ("F" or "O") arm during the Y-maze test in young and old C57B1/6 mice. Mice at 2 months of age were treated with IgG for 2 weeks (n ═ 18). 18-month-old mice were treated as follows: IgG treatment for 2 weeks (n ═ 18); compound 1 was treated at high dose for 2 weeks (n-15); compound 1 was treated at low dose for 2 weeks (n-31); or compound 1 was treated at low dose for 4 weeks (n ═ 16). Data shown are mean ± s.e.m., where P <0.05 and P < 0.01.

Figure 2B depicts the effect of compound 1 on the total number of visits by young and old C57B1/6 mice into the novel (N) arm versus the familiar (F) arm during the Y-maze test. Mice at 2 months of age were treated with IgG for 2 weeks (n ═ 18). 18-month-old mice were treated as follows: IgG treatment for 2 weeks (n ═ 18); compound 1 was treated at high dose for 2 weeks (n-15); compound 1 was treated at low dose for 2 weeks (n-31); or compound 1 was treated at low dose for 4 weeks (n ═ 16). Data shown are mean ± s.e.m., where P <0.05, P <0.01, P < 0.001.

Figure 3 depicts the effect of compound 1 on the round trip ("visit") of C57B1/6 mice into the novel arm and the familiar arm in the Y-maze test. The number of visits to each arm was plotted for each treatment group and paired t-assays were performed. 3-month-old mice (n ═ 15) received a subcutaneous infusion (0.5 μ L/hour) of vehicle (veh) by means of an Alzet minipump for four weeks, with one change. 16.5 month old mice were replaced once by subcutaneous infusion of vehicle (veh, n ═ 16) or 50mg/mL compound 1(n ═ 16) (0.5 μ L/hr) for four weeks via an Alzet micropump. Data shown are mean ± s.e.m.; p < 0.05.

FIG. 4A depicts the effect of Compound 1 on the average time it takes C57B1/6 mice to find a target hole in the Barnes maze test. The average time is plotted for each treatment group. Mice of 3 months of age (n ═ 18) received a subcutaneous infusion (0.5 μ L/hour) of vehicle (veh) by means of an Alzet minipump for four weeks, with one replacement. 16.5 month old mice (n-15) were subcutaneously infused (0.5 μ L/hr) with an Alzet mini-pump with vehicle (veh, n-15) or 50mg/mL compound 1 (n-15) for four weeks with one replacement. Data shown are mean ± s.e.m.

Figure 4B depicts the effect of compound 1 on the difference in latency for finding a target hole in the last and first trials in C57B1/6 mice on the last day of the barnes maze test. Differences were plotted for each treatment group and data were subjected to unpaired t-assays. 3-month-old mice (n ═ 18) received a subcutaneous infusion (0.5 μ L/hour) of vehicle (veh) by means of an Alzet minipump for four weeks, with one change. 16.5 month old mice were replaced once by subcutaneous infusion (0.5 μ L/hr) of vehicle (veh, n ═ 15) or 50mg/mL compound 1(n ═ 15) by Alzet micropump for four weeks. Data shown are mean ± s.e.m.

FIG. 5 depicts the effect of Compound 1 on neurogenesis in C57B1/6 mice, as determined by detecting BrdU-positive cells from all sections in the dentate gyrus. The number of BrdU positive cells from the dentate gyrus was plotted for each treatment group and the data was subjected to an unpaired t-assay with the 16.5-month age group. 3-month-old mice (n ═ 17) received a subcutaneous infusion (0.5 μ L/hour) of vehicle (veh) by means of an Alzet minipump for four weeks, with one change. 16.5 month old mice were replaced once by subcutaneous infusion (0.5 μ L/hr) of vehicle (veh, n ═ 17) or 50mg/mL compound 1(n ═ 17) by Alzet micropump for four weeks. Data shown are mean ± s.e.m.; p < 0.05.

Figure 6 depicts the levels of compound 1 (levels below 10nM) detected in the CSF of young (n-2) and old (n-3) C57BL/6 mice. Levels of compound 1 were detected using mass spectrometry.

Figure 7 depicts the distribution of compound 1 in C57BL/6JOlaHsd mouse tissues (measured as area under the curve (AUC)) as a function of time. Compounds were labeled with carbon-14 label and measurements were taken at 1, 24 and 168 hours.

Figure 8 depicts the pharmacokinetic profile of compound 1 after oral (P.O.) administration. Compound 1 was measured at 20 minutes, 2 hours, 8 hours and 12 hours post-dose from plasma from male 2-month-old C57Bl/6 mice that received 30mg/kg or 150mg/kg by oral gavage. Plasma concentrations are plotted as a function of time after administration.

Figure 9 depicts the effect of compound 1 on the search preference of young C57B1/6 mice during the open space test. Exploration preferences were plotted as the ratio of time spent in the periphery vs. center and the data were subjected to one-factor ANOVA. 2-month-old mice were treated as follows: twice daily oral vehicle (n-11); compound 1(n ═ 12) at 30mg/kg orally twice daily; twice daily oral vehicle plus intraperitoneal administration of recombinant mouse CCL11 (eotaxin-1, or "rmE") (n-14); or compound 1 aggravated group mouse CCL11 (n-14) administered orally at 30mg/kg twice daily. Data shown are mean ± s.e.m.; p < 0.01.

FIG. 10A depicts the effect of Compound 1 on the novel arm of the Y-maze test versus the time spent in the familiar arm in young C57B1/6 mice. The time spent in the novel arm versus the familiar arm was plotted for each treatment group and the data was paired for t-assays. 2-month-old mice were treated as follows: twice daily oral vehicle (n-13); compound 1(n ═ 14) at 30mg/kg orally twice daily; twice daily oral vehicle plus intraperitoneal administration of recombinant mouse CCL11 (eotaxin-1) (n 15); or compound 1-aggravated group mouse CCL11(n 15) was orally administered at 30mg/kg twice daily. Data shown are mean ± s.e.m.; p < 0.01.

Figure 10B depicts the effect of compound 1 on the ratio of time spent in the novel arm versus the familiar arm of the Y-maze test in young C57B1/6 mice. Ratios were plotted for each treatment group and the data subjected to ANOVA Kruskal-Wallis post hoc assays. 2-month-old mice were treated as follows: twice daily oral vehicle (n-12); compound 1(n ═ 14) at 30mg/kg orally twice daily; twice daily oral vehicle plus intraperitoneal administration of recombinant mouse CCL11 (eotaxin-1) (n 15); or compound 1-aggravated group mouse CCL11(n 15) was orally administered at 30mg/kg twice daily. Data shown are mean ± s.e.m.; p < 0.05.

Figure 10C depicts the effect of compound 1 on the ratio of the number of times a young C57B1/6 mouse entered the Y-maze test novel arm versus the familiar arm. Ratios were plotted for each treatment group and data was subjected to t-assays between vector and recombinant mouse CCL11 treatment. 2-month-old mice were treated as follows: twice daily oral vehicle (n-13); compound 1(n ═ 14) at 30mg/kg orally twice daily; twice daily oral vehicle plus intraperitoneal recombinant mouse CCL11 (eotaxin-1) (n 15); or compound 1-aggravated group mouse CCL11(n 15) was orally administered at 30mg/kg twice daily. Data shown are mean ± s.e.m.; p < 0.05.

Figure 11A depicts the effect of compound 1 on the mean latency for young C57B1/6 mice to find a target hole during the bahness maze test. The average latency is plotted in units of time for each trial for each treatment group. 2-month-old mice were treated as follows: twice daily oral vehicle (n-13); compound 1(n ═ 14) at 30mg/kg orally twice daily; twice daily oral vehicle plus intraperitoneal recombinant mouse CCL11 (eotaxin-1) (n 15); or compound 1-aggravated group mouse CCL11(n 15) was orally administered at 30mg/kg twice daily. Data shown are mean ± s.e.m.

Figure 11B depicts the effect of compound 1 on the latency time to find a target hole in young C57B1/6 mice during the barnes maze test. The mean latency of the last three trials was plotted for each treatment group and the data was subjected to unpaired t-assays. 2-month-old mice were treated as follows: twice daily oral vehicle (n-13); compound 1(n ═ 14) at 30mg/kg orally twice daily; twice daily oral vehicle plus intraperitoneal recombinant mouse CCL11 (eotaxin-1) (n 15); or compound 1-aggravated group mouse CCL11(n 15) was orally administered at 30mg/kg twice daily. Data shown are mean ± s.e.m.; p < 0.05.

Figure 12A depicts the effect of compound 1 on memory of novel arms in the Y-maze, by number of entries into novel arms versus total number of entries. 24-month-old mice were treated with 30mg/kg compound 1 or vehicle orally twice daily. Data shown are mean ± s.e.m.; p < 0.01.

Figure 12B depicts the effect of compound 1 on the total distance traveled in the Y-maze as a measure of autonomic activity. 24-month-old mice were treated with 30mg/kg compound 1 or vehicle orally twice daily. Data shown are mean ± s.e.m.; p < 0.05.

Figure 13A depicts the effect of compound 1 on the round trip ("number of visits") of C57B1/6 mice into the novel and familiar arms in a Y-maze test in 24-month-old mice. The number of visits to each arm was plotted for each treatment group and paired t-assays were performed. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Three weeks later, mice were subjected to the Y-maze test. Immediately prior to treatment, all mice received 50mg/kg IP BrdU injections for 5 consecutive days. Data shown are mean ± s.e.m.; novel arms were compared to familiar arms by paired Student's t-assay,. P <0.05,. P <0.01,. P < 0.001.

Figure 13B depicts the effect of compound 1 on the ratio of the number of entries of the novel arm versus the familiar arm for the entry into the Y-maze test for 24-month-old mice. Ratios were plotted for each treatment group and data was subjected to t-assays between vehicle and compound 1 treatment. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Three weeks later, mice were subjected to the Y-maze test. Immediately prior to treatment, all mice received 50mg/kg IP BrdU injections for 5 consecutive days. Data shown are mean ± s.e.m.; p <0.05, P <0.01, P <0.001 by Student's t-assay compared to controls.

Figure 13C depicts the effect of compound 1 on the time spent in the novel arm versus the familiar arm of the Y-maze test in 24-month-old C57B1/6 mice. The time spent in the novel arm versus the familiar arm was plotted for each treatment group and the data was paired for t-assays. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Three weeks later, mice were subjected to the Y-maze test. Immediately prior to treatment, all mice received 50mg/kg IP BrdU injections for 5 consecutive days. Data shown are mean ± s.e.m.; the novel arm was identified by paired Student's t-familiar arm,. P <0.05,. P <0.01,. P < 0.001.

Figure 13D depicts the effect of compound 1 on the ratio of time spent in the novel arm versus the familiar arm ("duration") of the Y-maze test in 24-month-old C57B1/6 mice. Ratios were plotted for each treatment group and the data subjected to t-assay. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Three weeks later, mice were subjected to the Y-maze test. Immediately prior to treatment, all mice received 50mg/kg IP BrdU injections for 5 consecutive days. Data shown are mean ± s.e.m.; p <0.05, P <0.01, P <0.001 by Student's t-assay compared to controls.

Figure 13E depicts the effect of compound 1 on the mean velocity of 24-month old C57B1/6 mice during the Y-maze test. The average speed was plotted for each treatment group and the data was subjected to t-test. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Three weeks later, mice were subjected to the Y-maze test. Immediately prior to treatment, all mice received 50mg/kg IP BrdU injections for 5 consecutive days. Data shown are mean ± s.e.m.; p <0.05, P <0.01, P <0.001 by Student's t-assay compared to controls.

Figure 14A depicts the effect of compound 1 on the average time it took for a 24-month old C57B1/6 mouse to find a target hole in the bahness maze test. The average time is plotted for each treatment group. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Three weeks later, mice were subjected to the Barnesian maze test. Data shown are mean ± s.e.m.; p <0.05 by Student's t-assay compared to control.

Figure 14B depicts the effect of compound 1 on the speed of 24-month old C57B1/6 mice in the bahness maze test, averaged over all trials in each treatment group. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Three weeks later, mice were subjected to the Barnesian maze test. Data shown are mean ± s.e.m.; p <0.05 by Student's t-assay compared to control.

Figure 15A depicts the effect of compound 1 on distance traveled in the open space test in 24-month-old C57B1/6 mice. The average travel distance is plotted for each treatment group. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Three weeks later, mice were subjected to open space testing. Data shown are mean ± s.e.m.

Figure 15B depicts the effect of compound 1 on the velocity of 24-month old C57B1/6 mice in the open space test. The average speed of the mice was plotted for each treatment group. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Three weeks later, mice were subjected to open space testing. Data shown are mean ± s.e.m.

Figure 16A depicts the effect of compound 1 on TNF α cytokine levels in the plasma of 24-month old C57B1/6 mice. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Inflammatory cytokine levels tend to be lower in mice treated with compound 1 than in vehicle-treated mice.

Figure 16B depicts the effect of compound 1 on IL6 cytokine levels in the plasma of 24-month old C57B1/6 mice. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Inflammatory cytokine levels tend to be lower in mice treated with compound 1 than in vehicle-treated mice.

Figure 16C depicts the effect of compound 1 on IL1 β cytokine levels in the plasma of 24-month old C57B1/6 mice. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Inflammatory cytokine levels tend to be lower in mice treated with compound 1 than in vehicle-treated mice.

Figure 16D depicts the effect of compound 1 on IL5 cytokine levels in the plasma of 24-month old C57B1/6 mice. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Inflammatory cytokine levels tend to be lower in mice treated with compound 1 than in vehicle-treated mice.

Figure 16E depicts the effect of compound 1 on IL17 cytokine levels in the plasma of 24-month old C57B1/6 mice. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. Inflammatory cytokine levels tend to be lower in mice treated with compound 1 than in vehicle-treated mice.

Figure 17 depicts the effect of compound 1 on activated microglia in 24-month old C57B1/6 mice. 23-month-old mice were subcutaneously administered either vehicle control or compound 1BID (twice daily) for 21 days. CD68+ activated microglia levels tended to be lower in compound 1-treated mice than in vehicle-treated mice.

Figure 18 depicts the effect of compound 1 on the percentage of eosinophils in complete White Blood Cell (WBC) counts in 24-month-old C57BI/6 mice treated for 3 weeks with control saline or 30mg/kg compound 1BID SQ. Compound 1 reduced endogenous age-related increases in blood eosinophils.

Figure 19 depicts the effect of compound 1 on the percentage of eosinophils in complete White Blood Cell (WBC) counts in 3-month-old hairless mice treated with control saline or 30mg/kg compound 1BID PO for 2 weeks. Compound 1 reductionOxazolone-induced increase in blood eosinophils.

Figure 20 shows the results of compound 1 on motor coordination in a rotarod test. 24-month-old C57 mice were treated by osmotic pump with continuous infusion of compound 1 or vehicle for 4 weeks. The treated mice were more successful in the two tests than the vehicle-treated mice, P < 0.05.

Figure 21 shows the results of compound 1 memory to the T-maze test. 24-month-old C57 mice were treated by osmotic pump with continuous infusion of compound 1 or vehicle for 4 weeks. The treated mice were more successful in the two tests than the vehicle-treated mice, P < 0.05.

Figure 22A shows the results of compound 1 on overnight fecal output. 24-month-old C57 mice were treated by osmotic pump with continuous infusion of compound 1 or vehicle for 4 weeks. The weight of overnight fecal pellets was measured. By Student's t-assay, compound 1-treated mice had significantly lower fecal output compared to vehicle-treated mice,. P < 0.05.

Figure 22B shows the results of compound 1 versus overnight water intake. 24-month-old C57 mice were treated by osmotic pump with continuous infusion of compound 1 or vehicle for 4 weeks. Total overnight water intake was measured. By Student's t-assay, compound 1-treated mice drunk significantly more water than vehicle-treated mice,. P < 0.05.

Figure 22C shows the results of compound 1 versus overnight food intake. 24-month-old C57 mice were treated by osmotic pump with continuous infusion of compound 1 or vehicle for 4 weeks. Total overnight food intake was measured. There was no difference in total overnight food intake between the two groups.

Figure 23 depicts the effect of compound 1(Cmpd 1) on the number of CD68 positive (CD68+) activated microglia in the brain of 3 month old mice treated with LPS and 18 days treated with compound 1. Compound 1 treated mice exhibited reduced CD68+ immunoreactivity, and thus reduced gliosis.

Fig. 24A and 24B depict the effect of compound 1 on anxiety in an open space assay in 3 month old mice treated with LPS IP for 4 weeks and compound 1 orally BID (twice daily) for 1 week. LPS treatment increased preference for the open space periphery, indicating increased anxiety. Compound 1 treatment reduced LPS-induced anxiety in open space. Data shown are mean ± s.e.m; p <0.05 by Student's t-assay.

Fig. 25A and 25B depict the effect of compound 1 on the number of entries into the novel and familiar arms in the Y-maze test in LPS-treated 3-month-old C57B1/6 mice. The number of visits to each arm was plotted for each treatment group and paired t-assays were performed. Mice at 3 months of age were dosed for 6 weeks with vehicle or LPS IP and orally for 3 weeks with vehicle control or compound 1BID (twice daily). Compound 1-treated mice showed significant preference for novel arms, while vehicle-treated mice did not. Data shown are mean ± s.e.m.; novel arms were compared to familiar arms by paired Student's t-assay,. P <0.01,. P < 0.05.

Fig. 26A and 26B depict the effect of compound 1 on IL1 β mRNA in the brain from LPS and/or compound 1 treated 3-month old C57B1/6 mice. Mice were dosed with vehicle control or LPS IP for 7 weeks and orally with vehicle or compound 1BID (twice daily) for 4 weeks. Tissues were harvested and RNA was prepared from cortical brain tissue. The level of IL1 β mRNA was measured by qPCR and data was provided relative to the vector control. There was a tendency for increased IL1 β mRNA levels with LPS treatment and a tendency for significant reduction with compound 1 treatment. Data shown are mean ± s.e.m; p <0.05 by Student's t-assay.

Figure 27A, figure 27B and figure 27C depict the effect of compound 1 on microglial activation in hippocampus from LPS and/or compound 1-treated 3-month-old C57B1/6 mice. Mice were dosed with vehicle or LPS IP for 7 weeks and orally with vehicle control or compound 1BID (twice daily) for 4 weeks. Tissues were harvested and brain sections were immunohistochemically for CD68 (a marker for activated microglia). There was a tendency for increased CD68 levels with LPS treatment and a strong tendency for decreased treatment with compound 1. Data shown are mean ± s.e.m.

Figure 28A, figure 28B and figure 28C depict the effect of compound 1 on total microglia in hippocampus from LPS and/or compound 1-treated 3-month-old C57B1/6 mice. Mice were dosed with vehicle control or LPS IP for 7 weeks and orally with vehicle or compound 1BID (twice daily) for 4 weeks. Tissues were harvested and brain sections were immunohistochemically against Iba1 (a marker for microglia). Treatment with LPS increased Iba1 significantly, and there was a reduced tendency for treatment with compound 1. Data shown are mean ± s.e.m.; p <0.001, P <0.05 by Student's t-assay.

Fig. 29A and 29B depict the effect of compound 1 on total astrocytes in hippocampus from 3-month-old C57B1/6 mice treated with LPS and/or compound 1. Mice were dosed with vehicle or LPS IP for 7 weeks and orally with vehicle or compound 1BID (twice daily) for 4 weeks. Tissues were harvested and brain sections were immunohistochemically targeted to GFAP (a marker for astrocytes). There was a reduced tendency to treat with compound 1. Data shown are mean ± s.e.m.

Figure 30 depicts the dosing regimen for three groups of C57BL/6 mice treated with the following method: (1) controls against LPS and compound 1; (2) control of LPS-loaded vehicle against compound 1; or (3) LPS plus Compound 1. All three groups were treated with control, LPS or compound 1 simultaneously for 3 consecutive days. The figure shows the results of histological analysis of mouse brain in an acute model of LPS-induced inflammation. Microglial proliferation was measured by determining the percentage of Iba-1 positive area in hippocampus. Data shown are mean ± s.e.m; p <0.05, P < 0.001; statistical significance was tested using a common one-way anova and Dunnett's multiple post-hoc comparisons between treatment groups.

Fig. 31 depicts the dosing regimen for three groups of C57BL/6 mice treated with the following method: vehicle control; vehicle-treated LPS; or compound 1. The figure shows the results of histological analysis of mouse brain in an acute model of LPS-induced inflammation. Microglial proliferation was measured by determining the percentage of Iba-1 positive area in hippocampus. Data shown are mean ± s.e.m; p <0.05, P < 0.0001; statistical significance was tested using a common one-way anova and Dunnett's multiple post-hoc comparisons between treatment groups.

Fig. 32 depicts a treatment study schedule for the mouse MPTP model of parkinson's disease. Two groups were studied; the first group tested gait and fine motor function 10 days after treatment, while the second group tested immune cell infiltration 3 days after treatment.

Fig. 33A shows a correlation heatmap depicting the degree of correlation of 97 walking parameters in gait and fine movement tests on study day 11 in C57Bl/6J mice 10 days after compound 1 treatment. 10 principal components are presented showing how the original parameters are related in the dataset. The greater the intensity of each parameter, the greater the contribution of that parameter in the corresponding principal component. Relative to the 10 individual principal components (x-axis), red is positively correlated and blue is negatively correlated. The left y-axis shows the overall grouping of single variables on the right y-axis. For example, "ILC" refers to the intralimb coordinates and "caudal B" refers to the caudal base.

Figure 33B shows fine motor ability and gait characteristics of C57Bl/6J mice at study day 11 after 10 days of compound 1 treatment. The overall gait analysis score of MPTP-treated C57Bl/6J mice is exemplified. Differences between treatment groups for each of the 10 principal components were pooled into composite scores and shown to be different from vehicle-treated controls. There was a significant difference in overall gait with MPTP treatment (. p <0.05) and no significant difference was present with compound 1 treatment.

Figure 34 depicts the anterior paw toe gap (a gait property evaluated in principal component analysis) at study day 11 of C57Bl/6J mice 10 days after compound 1 treatment. Data are presented as mean + SEM (group 1: vehicle + vehicle, n 15; group 2: MPTP + vehicle, n 14; group 3: MPTP + compound 1(30mg/kg), n 13). Statistical significance: p <0.05, group 2: MPTP + vehicle control group 1: vehicle + vehicle (unpaired t-test).

Figure 35 depicts the swing velocity of the forepaws (a gait property evaluated in principal component analysis) at study day 11 of C57Bl/6J mice 10 days after compound 1 treatment. Data are presented as mean + SEM (group 1: vehicle + vehicle, n 15; group 2: MPTP + vehicle, n 14; group 3: MPTP + compound 1(30mg/kg), n 13). Statistical significance: p <0.05, group 2: MPTP + vehicle control group 1: vehicle + vehicle (unpaired t-test).

Figure 36 depicts ankle range of motion (a gait property evaluated in principal component analysis) in C57Bl/6J mice at study day 11 after 10 days of compound 1 treatment. Data are presented as mean + SEM (group 1: vehicle + vehicle, n 15; group 2: MPTP + vehicle, n 14; group 3: MPTP + compound 1(30mg/kg), n 13). Statistical significance: p <0.05, group 2: MPTP + vehicle control group 1: vehicle + vehicle (unpaired t-test).

Figure 37 reports the acute effect of MPTP and compound 1 on T cell trafficking into the brain 3 days after compound 1 treatment. The total number of CD3 positive T cells counted in the substantia nigra of 3 30 μm sections from each mouse is presented. Data shown are mean ± s.e.m; p < 0.001; single factor analysis of variance, Sidak post hoc multiple comparison test.

Fig. 38A and 38B report the acute effect of MPTP and compound 1 on microglial hyperplasia 3 days after compound 1 treatment. Figure 38A shows the extent of CD68 positive area measured in the striatum (n-7 for saline + vehicle; n-8 for MPTP + vehicle; n-7 for MPTP + compound 1). Fig. 38B shows the extent of CD68 positive area measured in the substantia nigra dense section (n-7 for saline + vehicle; n-8 for MPTP + compound 1). Data shown are mean ± s.e.m; p <0.01, P <0.001, P < 0.0001; single factor analysis of variance, Sidak post hoc multiple comparison test.

FIG. 39 depicts plasma eotaxin-1 levels in 6-month old Line 61 synuclein-overexpressing transgenic mice. Eotaxin-1 levels (CCL11) in non-transgenic (NTg) versus transgenic (Tg) Line 61 synuclein mice were plotted in pg/mL concentrations.

FIG. 40 reports the results of steel wire hanging tests on Line 61 synuclein mice (transgenic (Tg) and non-transgenic (nTg) age-matched littermates). The average steel wire hang time for each group is shown, with animals from group C (non-transgenic, vehicle treated) exhibiting significantly higher steel wire hang times. Animals from group a (transgenic, compound 1-treated) exhibited significantly higher steel wire hang times than group B (transgenic, vehicle-treated). Data are shown as mean + SEM of all animals per group; p < 0.001; post-Dunn test; mann Whitney test P <0.05) for group a versus group B).

FIG. 41 reports the results of grip strength testing of Line 61 synuclein mice (transgenic (Tg) and non-transgenic (nTg) age-matched littermates). The average maximum grip g of each group is shown. Animals from groups a and C (transgenic, compound 1 treated and non-transgenic, respectively, vehicle treated) showed significantly higher grip strength compared to group B (transgenic, vehicle treated). Data are shown as mean ± SEM of all animals per group. Comparing each group to vehicle-treated transgenic animals (group B); one-way anova followed by Bonferroni post hoc tests.

Figure 42 reports the number of mice from each treatment group that were able to successfully pass through the balance beam in the balance beam walk test. N-15 mice in group a, N-14 mice in group B and N-15 mice in group C (the groups depicted in figure 40). All mice were able to pass through the easiest balance beam in test 1, while none of the mice from treatment group B passed through the hardest balance beam in test 5 (test 1: 13mm rectangular balance beam; test 2: 10mm rectangular balance beam; test 3: 28mm cylindrical balance beam; test 4: 16mm cylindrical balance beam; test 5: 11mm cylindrical balance beam).

Figure 43 reports the results of five trials balancing wood walking glide for three groups of mice (group a, transgene treated with compound 1; group B, transgene treated with vehicle; and group C, non-transgene treated with vehicle). Only mice that pass completely through the balance beam were included in the analysis. The mean number of slips [ n ] for each group is shown, and each graph represents one test (1-5). Data are mean ± SEM of all animals per group. Comparing each group to group B; one-way anova followed by Bonferroni post hoc tests. Statistical data for trial 5 could not be performed because none of the mice from group B were able to cross the balance beam.

Fig. 44A and 44B report eosinophil counts from peripheral blood. Figure 44A reports the percentage of eosinophils (on all leukocytes) in peripheral blood from three groups of mice (Tg Cmpd 1 ═ transgene treated with compound 1; Tg Veh ═ transgene treated with vehicle; nTG Veh ═ non-transgene treated with vehicle). The data were compared by t-test. Figure 44B reports the absolute eosinophil count in peripheral blood from three groups of mice (Tg Cmpd 1 ═ transgene treated with compound 1; Tg Veh ═ transgene treated with vehicle; nTG Veh ═ non-transgene treated with vehicle). The data were compared by t-test.

Fig. 45A to 45G report the effect of compound 1 on neuroinflammation. Figure 45A reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and the CD68 positive area quantified in hippocampus of transgenic, compound 1-treated mice (n ═ 14, 12, and 15, respectively). Figure 45B reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and the CD68 positive area quantified in the striatum of transgenic, compound 1-treated mice (n ═ 15, 11, and 16, respectively). Figure 45C reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and transgenic, Iba-1 positive area quantified in hippocampus of compound 1-treated mice (n ═ 14, 13, and 16, respectively). Figure 45D reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and the quantified Iba1 positive area in the striatum of transgenic, compound 1-treated mice (n ═ 15, 11, and 16, respectively). Data are mean +/-s.e.m.; p < 0.05. Figure 45E reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and transgenic, GFAP positive areas quantified in hippocampus of compound 1-treated mice (n ═ 14, 13, and 15, respectively). Figure 45F reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and the quantified GFAP positive area in the striatum of transgenic, compound 1-treated mice (n ═ 15, 13, and 15, respectively). Figure 45G reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and transgene, Iba-1 positive area quantified in substantia nigra dense part of compound 1 treated mice. Data are mean +/-s.e.m.; p < 0.05.

Figures 46A and 46B report compound 1 versus non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and effects of circulating levels of IL-4 and IL-6 cytokines in transgenic, compound 1-treated mice (n ═ 14, 15, and 17, respectively). Figure 46A reports the measured IL-4 levels in terminal heart plasma of all three groups. Figure 46B reports the measured IL-6 levels in terminal heart plasma of all three groups. Data shown are mean +/-s.e.m.; p <0.05, P < 0.01; one-way anova, Dunnett's multiple comparison test after the event.

FIGS. 47A-47D show the effect of Compound 1 on EAE-induced markers in the cerebellum of C57BL/6 mice. EAE results in an increase in CD3 and CD8 positive infiltrating T cells in the cerebellum. Figure 47A shows an increase in CD3 positive infiltrating T cells in the cerebellum, which was significantly reduced after 9 days of treatment with compound 1. Fig. 47B shows an increase in CD8 positive infiltrating cytotoxic T cells in the cerebellum, which was significantly reduced after 9 days of treatment with compound 1. FIG. 47C shows a significant increase in Iba-1 positive area in cerebellum after EAE, which was significantly reduced in cerebellum after 9 days of treatment. Figure 47D shows a significant increase in CD68 positive area in cerebellum after EAE, which was significantly reduced in cerebellum after 9 days of treatment. Data shown are mean +/-s.e.m; p <0.05, P < 0.01; one-way anova, Dunnett's multiple comparison test after the event.

Figure 48 depicts the concentration of human eotaxin-1 in a proteomic study screen. The relative concentration of human eotaxin-1 was measured in a commercially available affinity-based assay (SomaLogic). Plasma samples from each of the 18, 30, 45, 55 and 66 year old donors were plotted.

Figure 49A depicts the effect of compound 1 on inhibition of eosinophil shape change. Whole blood from compound 1-treated humans was incubated with recombinant eotaxin to trigger eosinophil shape change. The inhibition of shape change was plotted against plasma compound 1 concentration.

Figure 49B depicts the effect of compound 1 on CCR3 internalization. Whole human blood from compound 1 treatment was incubated with recombinant eotaxin to trigger CCR3 internalization and labeled with anti-CCR 3 antibody. Inhibition of CCR3 internalization by compound 1 was plotted against plasma compound 1 concentration.

Detailed description of the invention VI

Aspects of the invention include methods of treating injury/neurodegenerative diseases associated with aging. Age-related damage can manifest in a number of different ways, for example, in cognitive and/or physiological damage associated with aging, for example, in the form of damage to central or peripheral organs of the body, such as, but not limited to: cell damage, tissue damage, organ dysfunction, age-related life shortening, and carcinogenesis, wherein specific organs and tissues of interest include, but are not limited to, skin, neurons, muscle, pancreas, brain, kidney, lung, stomach, intestine, spleen, heart, adipose tissue, testis, ovary, uterus, liver, and bone; and in a form that reduces neurogenesis, and the like.

In some embodiments, the aging-associated impairment is an impairment of cognitive ability associated with aging in the individual, i.e., an aging-associated cognitive impairment. Cognitive ability or "cognition" means mental processes that include attention and concentration, learning complex tasks and concepts, memory (short and/or long term acquisition, retention and retrieval of new information), information processing (processing of information collected by five senses), visual spatial functions (visual perception, depth perception, use of mental imagery, copying, object or shape construction), language production and understanding, verbal fluency (vocabulary discovery), problem solving, decision making, and function execution (planning and prioritization). By "cognitive decline" is meant a progressive decline in one or more of these abilities, e.g., decline in memory, language, thinking, judgment, and the like. By "cognitive impairment" and "cognitive impairment" is meant a decrease in cognitive ability relative to a healthy individual (e.g., an age-matched healthy individual) or relative to the individual's ability at an earlier time point (e.g., 2 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 2 years, 5 years, or 10 years or earlier). Cognitive impairment associated with aging includes impairment of cognitive ability commonly associated with aging, including, for example, cognitive impairment associated with the natural aging process, e.g., mild cognitive impairment (m.c.i.); and cognitive impairment associated with aging-related disorders, i.e., disorders that increase in frequency with aging, e.g., neurodegenerative disorders such as alzheimer's disease, parkinson's disease, dementia with lewy bodies, frontotemporal dementia, huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, and vascular dementia, and the like.

By "treating" is meant at least ameliorating one or more symptoms associated with the aging-related injury afflicting the subject, where amelioration is used in a broad sense to refer to a reduction in at least a parameter magnitude (e.g., symptoms associated with the injury being treated). Thus, treatment also includes situations in which the pathological condition, or at least the symptoms associated therewith, is completely inhibited (e.g., prevented from occurring) or halted (e.g., terminated), such that the adult mammal no longer suffers from the injury or at least the symptoms that characterize the injury. In some instances, "treating" refers to obtaining a desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of completely or partially preventing the disease or a symptom thereof and/or therapeutic in terms of a partial or complete cure of the disease and/or adverse effects due to the disease. "treatment" may be any treatment of a disease in a subject and includes: (a) preventing the occurrence of a disease in a subject, which subject may be predisposed to the disease but has not yet been diagnosed with the disease; (b) inhibiting the disease, i.e., inhibiting its development; or (c) ameliorating the disease, i.e., causing regression of the disease. Treatment can result in a variety of different physical manifestations, e.g., modulation of gene expression, increased neurogenesis, tissue or organ rejuvenation, and the like. In some embodiments, ongoing treatment of the disease occurs, wherein the treatment stabilizes or reduces the patient's undesirable clinical symptoms. These treatments may be performed before the affected tissue is completely dysfunctional. The targeted therapy may be administered during, and in some cases after, the symptomatic phase of the disease.

In some cases where the age-related impairment is age-related cognitive decline, treatment by the methods of the invention slows or reduces the progression of the age-related cognitive decline. In other words, the cognitive ability of the individual decreases (if at all) more slowly after treatment by the disclosed methods than before or without treatment by the disclosed methods. In some examples, treatment by the methods of the invention stabilizes the cognitive ability of the individual. For example, progression of cognitive decline in an individual suffering from age-related cognitive decline will cease following treatment by the disclosed methods. As another example, after treatment by the disclosed methods, cognitive decline in an individual (e.g., an individual 40 years of age or older) is prevented, which individual is expected to suffer from cognitive decline associated with aging. In other words, no (further) cognitive impairment was observed. In some examples, treatment by the methods of the invention reduces or reverses cognitive impairment, e.g., as observed by improving cognitive ability in individuals suffering from age-related cognitive decline. In other words, the cognitive ability of an individual suffering from age-related cognitive decline is better after treatment by the disclosed method than they were before treatment by the disclosed method, i.e., they are improved after treatment. In some examples, treatment by the methods of the invention abrogates cognitive impairment. In other words, after treatment by the disclosed methods, the cognitive ability of the individual suffering from age-related cognitive decline is restored to, for example, its level at about 40 years of age or less in the individual, e.g., as evidenced by an improvement in cognitive ability of the individual suffering from age-related cognitive decline.

In some cases where the age-related impairment is age-related movement impairment or decline, the progression of the age-related movement impairment or decline is slowed or reduced by treatment according to the methods of the invention. In other words, the exercise capacity of the individual decreases more slowly (if at all) after treatment by the disclosed method than before or without treatment by the disclosed method. In some examples, treatment by the methods of the invention stabilizes the motor capacity of the individual. For example, the progression of the decline in exercise in an individual suffering from age-related decline in exercise will cease following treatment by the disclosed methods. As another example, after treatment by the disclosed methods, a decline in exercise is prevented in an individual (e.g., an individual 40 years of age or older) who is expected to suffer from a decline in exercise associated with aging. In other words, no (further) motor impairment was observed. In some examples, treatment by the methods of the invention reduces or reverses motor impairment, e.g., as observed by improving motor coordination or ability in individuals suffering from age-related motor decline. In other words, the exercise capacity of an individual suffering from age-related exercise decline is better after treatment by the disclosed method than it was before treatment by the disclosed method, i.e., it is improved after treatment. In some examples, treatment by the methods of the invention eliminates sports injuries. In other words, after treatment by the disclosed methods, the motor coordination or ability of the subject suffering from the age-related motor decline is restored to, for example, their level at about 40 years of age or less in the subject, e.g., as evidenced by an improvement in the motor coordination or ability of the subject suffering from the age-related motor decline.

In some examples, treatment of an adult mammal according to these methods results in alteration of a central organ (e.g., a central nervous system organ, such as the brain, spinal cord, etc.), where the alteration can be manifested in a number of different ways, e.g., as described in more detail below, including but not limited to molecular, structural, and/or functional, e.g., in a form that enhances neurogenesis.

Methods of treating a dysfunction caused by a neurodegenerative disease are provided, the methods comprising administering a compound from the following formula. Embodiments of the invention include methods of improving cognitive or motor activity in a subject having a brain-related, cognitive-related, or motor disorder comprising administering a therapeutically effective amount of a compound from the following formula. Other embodiments of the invention include methods of increasing neurogenesis in a subject having a brain-related or cognition-related disorder comprising administering a therapeutically effective amount of a compound from the formula. Other embodiments of the invention include methods of soothing or treating symptoms of a brain-related or cognition-related disorder comprising administering a therapeutically effective amount of a compound from the formula below. Other embodiments of the invention include methods of soothing symptoms of a central nervous system related disorder comprising administering a therapeutically effective amount of a primary peripheral agent from the formula below. Other embodiments of the invention include methods of improving locomotor activity in a subject having age-related motor dysfunction, comprising administering a therapeutically effective amount of a compound from the formula. The methods of the invention may also include monitoring for improvement in age-related disorders, including, for example, improving cognition, motor activity, and neurogenesis, among others, in a subject diagnosed with one or more such disorders or dysfunctions.

Other embodiments of the invention include administering a therapeutically effective amount of a compound, wherein the compound is in the form of a co-crystal or salt of the formula. Other embodiments of the invention include administering a therapeutically effective amount of a compound, wherein the compound is in the form of an individual optical isomer, a mixture of individual enantiomers, a racemate or an enantiomerically pure compound. Other embodiments of the invention also include the administration of a therapeutically effective amount of a compound, wherein the compound is in the form of pharmaceutical compositions and formulations discussed further below.

Other embodiments of the invention for treating age-related movement impairment or decline include modifiers that inhibit the eotaxin/CCR 3 pathway. Such modifiers include not only compounds from the following formulae, but also other eotaxin and CCR3 inhibitors. Contemplated modifiers include by way of example and not limitation: compounds of the following formula and other CCR3 small molecule inhibitors (e.g., bispidine derivatives as described, for example, in U.S. patent No. 7,705,153, cyclic amine derivatives as described, for example, in US 7,576,117, and CCR3 antagonists as described in Pease JE and Horuk R, Expert Opin Drug Discov (2014)9(5):467-83, all of which are incorporated herein by reference in their entirety); anti-eotaxin antibodies; anti-CCR 3 antibodies; aptamers that inhibit eotaxin or CCR3 expression or function (methods of producing such aptamers include U.S. Pat. nos. 5,270,163, 5,840,867, 6,180,348); antisense oligonucleotides or sirnas that inhibit the expression or function of eotaxin or CCR3 (e.g., as described in U.S. patent No. 6,822,087); and soluble CCR3 receptor proteins (e.g., decoy), and the like.

Methods of use of or with diagnostic tests for the described age-related damage are also provided. One embodiment of such a diagnostic or companion diagnostic test is an in vitro diagnostic test. An embodiment of an in vitro diagnostic test is a companion device used with a particular therapeutic agent. Embodiments of particular therapeutic agents include, for example, but are not limited to, small molecule inhibitors of CCR3, anti-CCR 3 antibodies, small molecule inhibitors of the CCR3 ligand Eotaxin-1, anti-Eotaxin-1 antibodies, and antisense RNA to either Eotaxin-1 or CCR 3.

By way of example and not limitation, such diagnosis or companion diagnosis includes determining or detecting the presence of a subset of leukocytes from a subject. The diagnosis or companion diagnosis may also be a determination of the presence, relative or absolute concentration, relative or absolute number of eosinophils in a subject's blood, tissue or other such sample. Blood may be obtained, for example, by venipuncture or other similar methods. Other samples may include, for example, but are not limited to sputum, cerebrospinal fluid, or tissue biopsy.

Methods of detecting or determining the presence, absolute or relative concentration or relative or absolute number of leukocytes, such as eosinophils, neutrophils, lymphocytes, basophils, and monocytes, are known to those of ordinary skill in the art. Methods of determining the presence, absolute or relative concentration or relative or absolute number of eosinophils are also known to those of ordinary skill in the art. See Walsh GM, Eosinophils-Methods and Protocols ISBN:9781493910151, which is incorporated herein by reference in its entirety. These methods include, by way of example and not limitation: (1) 5-Classification Whole BLOOD cell counts using Hematology Analyzer (see, e.g., O' Neil et al, Laboratory Hematology 7: 116-; (3) flow cytometric quantification of eosinophils using antibodies specific for various immune cells such as CD45, CD125, CD193, F4/80, and Siglec-8(Abcam, san Francisco, Calif.) (see, e.g., Yu Y-RA et al, Am J Respir Cell Mol biol.2016, 1 month; 54(1): 13-24, and Cossariza et al, Eur.J.Immunol.201747: 1584-1797)); (4) flow Cytometry was used to quantify eosinophil autofluorescence (see, e.g., Guenther G et al, J Immunol May 1,2015,194(1Supplement)206.12 and Thuurau AM et al, Cytometry 23:159-58 (1996)); (5) isolation of eosinophils from whole blood using Dextran 70, Ficoll-Paque and antibody-based magnetic colloidal cell isolation (see, e.g., Munoz NM et al, Nature Protocols,2613-20 (2006); Akuthota P et al, Curr Protoc Immunol 98(1): pp.7.31.1-7.31.8,2012; and Akuthota P et al, Methods Mol Biol 1178:13-20 (2014)); (6) histological staining by general staining (eosin or similar dyes) or by antibody-based staining for Eosinophil-specific proteins (major basic protein (MBP), Eosinophil Cationic Protein (ECP), Eosinophil Peroxidase (EPO), Eosinophil-derived neurotoxin (EDN), SiglecF (or Siglec8)) (see, e.g., Koller DY et al, Allergy 54(10):1094-9(1999) and Akuthoa P., Capron K., Weller P.F, (2014) eosinophii Purification from Peripheral blood, in Walsh G. (eds.) eosinophiums, Methods in Molecular Biology (Methods and Protocols),1178 Vol. mana Press, New York, NY); (7) processes spatially associated with eosinophils (e.g., degranulation), eosinophil extracellular capture (EET), and/or histological staining of specific proteins therein (e.g., Galectin 10).

Diagnostic or companion diagnostic devices may incorporate these methods to determine the presence, absolute or relative concentration or relative or absolute number of eosinophils. The device may be used with other methods of the invention by determining the presence, absolute or relative concentration or relative or absolute number of eosinophils. By way of example and not limitation, these other methods may include methods of treating aging-related injuries/neurodegenerative diseases as described herein. In some embodiments, the aging-related impairment is an aging-related impairment in the cognitive ability of the individual, i.e., an aging-related impairment. Such age-related injuries may include, by way of example but not limitation, neurodegenerative diseases such as alzheimer's disease, parkinson's disease, dementia with lewy bodies, frontotemporal dementia, huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, vascular dementia, and the like. Such age-related impairments may include cognitive impairment and/or motor impairment or deterioration.

Embodiments of the invention include treating a subject diagnosed with age-related damage in combination with a therapeutically effective amount of at least one compound of the invention disclosed herein and a diagnostic or companion diagnostic device. Another embodiment of the invention comprises treating the subject with a therapeutically effective amount of compound 1 disclosed herein in combination with a diagnostic device. Embodiments of the device may be used, for example, to determine or detect the presence, absolute or relative concentration, absolute or relative amount of eosinophils in a blood or tissue sample of a subject. Another embodiment performs the step of detecting or determining the presence, absolute or relative concentration, or absolute or relative amount of eosinophils in a blood or tissue sample of the subject before, during or after treatment.

The experimental data herein show that mammalian models of parkinson's disease with synuclein overexpression result in eosinophilia depletion, which is restored to a level in non-transgenic mice treated with compound 1, suggesting a beneficial immunomodulatory effect in such models of parkinson's disease. This also suggests that determining eosinophil levels in parkinson's disease patients treated with compound 1 can be a biomarker for the disease, including determining the level of progression, arrest or regression of the disease as well as the therapeutic effect (see, e.g., fig. 44A and 44B).

Another embodiment performs the step of detecting or determining the presence, absolute or relative concentration, absolute or relative amount of eosinophils in a blood or tissue sample of a subject diagnosed with parkinson's disease before, during or after treatment. Another embodiment comprises determining the presence, absolute or relative concentration, or absolute or relative amount of eosinophils in a blood or tissue sample of a subject diagnosed with parkinson's disease prior to treatment with a compound of the invention, e.g., compound 1, to obtain a baseline concentration or baseline amount. Another embodiment then follows the step of detecting or determining the level of eosinophils after treatment to compare this level to a baseline concentration or amount of eosinophils to monitor the effect of the treatment. Comparing these levels may show an increase or decrease in the number or concentration of eosinophils compared to baseline. Another embodiment of the invention comprises the step of monitoring the progression of the disease or injury by comparing the number or concentration of eosinophils, wherein the progression of the disease is improved if the number increases after treatment. In another embodiment, the improvement may comprise an increase in the number or concentration of eosinophils in the blood or tissue of the subject of 1 to 5% from baseline; increase by 5 to 10%; increase by 11 to 15%; increase by 16 to 20%; increase by 21 to 25%; increase by 26 to 30%; increase by 31 to 35%; increase by 36 to 40%; increase by 41 to 45%; increase by 46 to 50%; increase by 51 to 55%; increase by 56 to 60%; increase by 61 to 65%, increase by 66 to 70%; increase by 71 to 75%; increase by 76 to 80%; increase 81 to 85%; increase 86 to 90%; increase by 91 to 95%; increase by 96 to 100%; and increasing 1 to 1.5 x, 1.5 to 2 x, 2 to 2.5 x, 2.5 to 3 x, 3 to 3.5 x, 3.5 to 4 x, 4 to 4.5 x, 4.5 to 5 x, 5 to 5.5 x, 5.5 to 6 x, 6 to 6.5 x, 6.5 to 7 x, 7 to 7.5 x, 7.5 to 8 x, 8 to 8.5 x, 8.5 to 9 x, 9 to 9.5 x, 9.5 to 10 x, and greater than 10 x.

Another embodiment of the invention comprises diagnosing Parkinson's disease by determining a baseline for the number or concentration of eosinophils in the blood or tissue of a subject and comparing the baseline to a standard number or concentration of eosinophils in a population without Parkinson's disease that has a normal eosinophil count. Eosinophil count is known in the art to be the number of eosinophils in vivo. For adults, the normal eosinophil count is 30 to 350 cubic millimeters (mm) in blood³) Up to 500 cubic millimeters (mm)³) (see Medical News Today, available in 2018 at 12: https:// www.medicalnewstoday.com/articules/323868. php, which is incorporated herein by reference in its entirety). Greater than 500mm in blood³The count of (a) is considered eosinophilia. Below normal eosinophil counts (supra) occur in some diseases, such as alcoholism and cortisol overprotection. One aspect of the invention detects or determines the number or concentration of eosinophils in a subject suspected of having Parkinson's disease. If the number or concentration of eosinophils in the subject is below normal (e.g., 30mm in blood) ³) The caregiver can use the results to determine a diagnosis of the disease.

a. Compound (I)

The methods of the invention further comprise administering to the subject the following compound. In groups (groups), radicals (radicals) or moieties (moities) as defined in the "compounds" section herein, the number of carbon atoms is generally indicated before the group, e.g.E.g. C_1-6Alkyl means an alkyl group or alkyl group having 1 to 6 carbon atoms. In general, for groups containing two or more subgroups disclosed in this section "compounds", the last named group is the point of attachment of the group, e.g., "sulfanyl" means a monovalent group of the formula HS-Alk-. Unless otherwise indicated below, known definitions of term control and known stable valencies are assumed and achieved in all formulae and groups.

Embodiments of the invention also include administering to a subject a compound of formula 1, wherein

A is CH₂O or N-C_1-6-an alkyl group;

R¹is selected from

·NHR^1.1、NMeR^1.1；

·NHR^1.2、NMeR^1.2；

·NHCH₂-R^1.3；

·NH-C_3-6-cycloalkyl, wherein optionally one carbon atom is replaced by a nitrogen atom, wherein the ring is optionally substituted by a group selected from C_1-6Alkyl, O-C_1-6Alkyl, NHSO₂Phenyl, NHCONH-phenyl, halogen, CN, SO₂-C_1-6-alkyl and COO-C_1-6-one or two residue substitutions of alkyl;

·C_{9 or 10}-bicyclic ring, wherein one or two carbon atoms are replaced by nitrogen atoms and the ring system is bonded to the basic structure of formula 1 via nitrogen atoms and wherein the ring system is optionally selected from C _1-6-alkyl, COO-C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, NO₂Halogen, CN, NHSO₂-C_1-6-alkyl, methoxy-phenyl substituted with one or two residues;

selected from NHCH (pyridyl) CH₂COO-C_1-6Alkyl, NHCH (CH)₂O-C_1-6-alkyl) -benzimidazolyl radicalA group, optionally substituted with halogen or CN; or

1-aminocyclopentyl, optionally methyl-Oxadiazole substitution;

R^1.1is phenyl, optionally via a radical selected from C_1-6Alkyl radical, C_2-6-alkenyl, C_2-6-alkynyl, C_1-6-haloalkyl group, C_1-6alkylene-OH, C_2-6-alkenylene-OH, C_2-6-alkynylene-OH, CH₂CON(C_1-6-alkyl groups)₂、CH₂NHCONH-C_3-6-cycloalkyl, CN, CO-pyridyl, CONR^1.1.1R^1.1.2、COO-C_1-6Alkyl, N (SO)₂-C_1-6-alkyl) (CH₂CON(C_1-4-alkyl groups)₂)O-C_1-6-alkyl, O-pyridyl, SO₂-C_1-6Alkyl, SO₂-C_1-6alkylene-OH, SO₂-C_3-6-cycloalkyl, SO₂Piperidinyl, SO₂NH-C_1-6Alkyl, SO₂N(C_1-6-alkyl groups)₂Halogen, CN, CO-morpholinyl and CH₂-one or two residues of pyridyl optionally substituted by a group selected from C_1-6-alkyl, NHC_1-6-alkyl and a heterocycle substituted with one or two residues of ═ O;

R^1.1.1is H, C_1-6Alkyl radical, C_3-6-cycloalkyl, C_1-6-haloalkyl, CH₂CON(C_1-6-alkyl groups)₂、CH₂CO-Azacyclobutyl, C_1-6alkylene-C_3-6-cycloalkyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, C_1-6alkylene-OH or thiadiazolyl, optionally via C_1-6-alkyl substitution;

R^1.1.2Is H, C_1-6Alkyl, SO₂C_1-6-an alkyl group;

or R^1.1.1And R^1.1.2Together form a quaternary, five-or six-membered carbocyclic ring, optionally containing substitution of theOne N or O of a ring carbon atom, and optionally via C_1-6Alkyl radical, C_1-4-alkylene-OH, OH and ═ O substituted with one or two residues;

or

R^1.1Is phenyl, in which two adjacent residues together form a five-or six-membered carbocyclic aromatic or nonaromatic ring, optionally containing one or two N, S or SO independently of one another₂Replacing a carbon atom of said ring, wherein said ring is optionally substituted by C_1-4-alkyl or ═ O substitution;

R^1.2is selected from

Heteroaryl, optionally via a radical selected from C_1-6Alkyl radical, C_2-6-alkenyl, C_2-6-alkynyl, C_3-6-cycloalkyl, CH₂COO-C_1-6Alkyl, CONR^1.2.1R^1.2.2、COR^1.2.3、COO-C_1-6Alkyl, CONH₂、O-C_1-6Alkyl, halogen, CN and SO₂N(C_1-6-alkyl groups)₂By one or two residues of (A), or R^1.2Is optionally selected from C_1-6-heteroaryl substituted with one or two residues of alkyl;

heteroaryl, optionally substituted by a five-or six-membered carbocyclic non-aromatic ring, containing, independently of one another, two N, O, S or SO groups replacing carbon atoms on the ring₂；

Aromatic or non-aromatic C_{9 or 10}-bicyclic, in which one or two carbon atoms are replaced by N, O or S, each of which is optionally substituted by a group selected from N (C)_1-6-alkyl groups)₂、CONH-C_1-6-alkyl and one or two residue substitutions of ═ O;

A heterocyclic non-aromatic ring, optionally substituted by pyridyl;

4, 5-dihydro-naphtho [2,1-d ]]Thiazoles, optionally via NHCO-C_1-6-an alkyl substitution,

R^1.2.1H、C_1-6alkyl radical, C_1-6alkylene-C_3-6-cycloalkyl, C_1-4Alkylene-phenyl, C_1-4Alkylene-furyl radical, C_3-6-cycloalkyl, C_1-4alkylene-O-C_1-4Alkyl radical, C_1-6-haloalkyl orFive-or six-membered carbocyclic non-aromatic ring optionally containing, independently of each other, one or two N, O, S or SO replacing carbon atoms on the ring₂And optionally substituted with 4-cyclopropylmethyl-piperazinyl;

R^1.2.2is H, C_1-6-an alkyl group;

R^1.2.3is a five-or six-membered carbocyclic non-aromatic ring, optionally containing, independently of one another, one or two N, O, S or SO replacing carbon atoms on the ring₂；

R^1.3Selected from phenyl, heteroaryl or indolyl, each of which is optionally selected from C_1-6Alkyl radical, C_3-6-cycloalkyl, O-C_1-6Alkyl, O-C_1-6-one or two residue substitutions of haloalkyl, phenyl and heteroaryl;

R²is selected from C_1-6Alkylene-phenyl, C_1-6-alkylene-naphthyl, and C_1-6-alkylene-heteroaryl; each of which is optionally selected from C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, O-C_1-6-one, two or three residue substitutions of haloalkyl and halogen;

R³is H, C_1-6-an alkyl group;

R⁴is H, C_1-6-an alkyl group;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the invention also includes administering to a subject a compound of formula 1 (above), wherein

A is CH₂O or N-C_1-4-an alkyl group;

R¹is selected from

·NHR^1.1、NMeR^1.1；

·NHR^1.2、NMeR^1.2；

·NHCH₂-R^1.3；

R^1.1Is phenyl, optionally via a radical selected from C_1-6Alkyl radical, C_2-6-alkenyl, C_2-6-alkynyl, C_1-6Haloalkyl, C_1-6Alkylene (E) sradical-OH, C_2-6-alkenylene-OH, C_2-6-alkynylene-OH, CH₂CON(C_1-6-alkyl groups)₂、CH₂NHCONH-C_3-6-cycloalkyl, CN, CO-pyridyl, CONR^1.1.1R^1.1.2、COO-C_1-6Alkyl, N (SO)₂-C_1-6-alkyl) (CH₂CON(C_1-4-alkyl groups)₂)O-C_1-6-alkyl, O-pyridyl, SO₂-C_1-6Alkyl, SO₂-C_1-6alkylene-OH, SO₂-C_3-6-cycloalkyl, SO₂Piperidinyl, SO₂NH-C_1-6Alkyl, SO₂N(C_1-6-alkyl groups)₂Halogen, CN, CO-morpholinyl, CH₂-substitution of one or two residues of pyridyl, or R^1.1Is optionally selected from C_1-6-alkyl, NHC_1-6-alkyl, ═ O, and heterocycles substituted with one or two residues;

R^1.1.1is H, C_1-6Alkyl radical, C_3-6-cycloalkyl, C_1-6Haloalkyl, CH₂CON(C_1-6-alkyl groups)₂、CH₂CO-Azacyclobutyl, C_1-6alkylene-C_3-6-cycloalkyl, CH₂-pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, C_1-6alkylene-OH or thiadiazolyl, optionally via C_1-6-alkyl substitution;

R^1.1.2is H, C_1-6Alkyl, SO₂C_1-6-an alkyl group;

or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one N or O replacing a carbon atom on the ring, optionally via a substituent selected from C_1-6Alkyl radical, C_1-4-alkylene-OH, ═ O, substituted with one or two residues;

Or

R^1.1Is phenyl, wherein two adjacent residues together form a five-or six-membered carbocyclic aromatic or non-aromatic ring, optionally containing, independently of each other, one or two N, S or SO replacing a carbon atom on the ring₂Wherein the ring is optionally substituted by C_1-4-alkyl or ═ O substitution;

R^1.2is selected from

Heteroaryl, optionally via a radical selected from C_1-6Alkyl radical, C_2-6-alkenyl, C_2-6-alkynyl, C_3-6-cycloalkyl, CH₂COO-C_1-6Alkyl, CONR^1.2.1R^1.2.2、COR^1.2.3、COO-C_1-6Alkyl, CONH₂、O-C_1-6Alkyl, halogen, CN, SO₂N(C_1-4-alkyl groups)₂Or optionally substituted by one or two residues selected from C_1-6-heteroaryl substituted with one or two residues of alkyl;

heteroaryl, optionally substituted by a five-or six-membered carbocyclic non-aromatic ring, containing, independently of one another, two N, O, S or SO replacing carbon atoms on the ring₂；

R^1.2.1Is H, C_1-6Alkyl radical, C_1-6alkylene-C_3-6-cycloalkyl, C_1-4Alkylene-phenyl, C_1-4Alkylene-furyl radical, C_3-6-cycloalkyl, C_1-4alkylene-O-C_1-4Alkyl radical, C_1-6Haloalkyl or a five-or six-membered carbocyclic non-aromatic ring, optionally containing, independently of one another, one or two N, O, S or SO replacing carbon atoms on the ring₂And optionally substituted with 4-cyclopropylmethyl-piperazinyl;

R^1.2.2is H, C_1-6-an alkyl group;

R^1.2.3is a five-or six-membered carbocyclic non-aromatic ring, optionally containing, independently of one another, one or two N, O, S or SO replacing carbon atoms on the ring ₂；

R^1.3Selected from phenyl, heteroaryl or indolyl, each optionally via C_1-6Alkyl radical, C_3-6-cycloalkyl, O-C_1-6Alkyl, O-C_1-6-one or two residue substitutions of haloalkyl, phenyl, heteroaryl; wherein in some instances R¹3 is selected from phenyl, pyrazolyl, isoAzolyl, pyridyl, pyrimidinyl, indolyl orOxadiazolyl, each of which is optionally selected from C_1-6Alkyl radical, C_3-6-cycloalkyl, O-C_1-6Alkyl, O-C_1-6-one or two residue substitutions of haloalkyl, phenyl, pyrrolidinyl;

R²is selected from C_1-6Alkylene-phenyl, C_1-6-alkylene-naphthyl, and C_1-6-alkylene-thiophenyl; each of which is optionally selected from C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, O-C_1-6-one, two or three residue substitutions of haloalkyl and halogen;

R³is H, C_1-4-an alkyl group;

R⁴is H, C_1-4-an alkyl group;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1 (above), wherein

A is CH₂O or N-C_1-4-an alkyl group;

R¹is selected from

·NHR^1.1、NMeR^1.1；

R^1.1Is phenyl, optionally via a radical selected from C_1-6Alkyl radical, C_2-6-alkenyl, C_2-6-alkynyl, C_1-6-haloalkyl group, C_1-6alkylene-OH, C_2-6-alkenylene-OH, C_2-6-alkynylene-OH, CH₂CON(C_1-6-alkyl groups)₂、CH₂NHCONH-C_3-6-cycloalkyl, CN, CO-pyridyl, CONR^1.1.1R^1.1.2、COO-C_1-6Alkyl, N (SO) ₂-C_1-6-alkyl) (CH₂CON(C_1-4-alkyl groups)₂)O-C_1-6-alkyl, O-pyridyl, SO₂-C_1-6Alkyl, SO₂-C_1-6alkylene-OH, SO₂-C_3-6-cycloalkyl, SO₂Piperidinyl, SO₂NH-C_1-6Alkyl, SO₂N(C_1-6-alkyl groups)₂Halogen, CN, CO-morpholinyl, CH₂-substitution of one or two residues of pyridyl, or R^1.1Is optionally selected from C_1-6-alkyl, NHC_1-6-alkyl, ═ O, and heterocycles substituted with one or two residues;

R^1.1.1is H, C_1-6Alkyl radical, C_3-6-cycloalkyl, C_1-6-haloalkyl, CH₂CON(C_1-6-alkyl groups)₂、CH₂CO-Azacyclobutyl, C_1-6alkylene-C_3-6-cycloalkyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, C_1-6alkylene-OH or thiadiazolyl, optionally via C_1-6-alkyl substitution;

R^1.1.2is H, C_1-6Alkyl, SO₂C_1-6-an alkyl group;

or R^1.1.1And R^1.1.2Together forming a four-, five-or six-membered carbocyclic ring, optionally replacing a carbon atom of the ring, containing one N or O, optionally via a substituent selected from C_1-6Alkyl radical, C_1-4-alkylene-OH, ═ O, substituted with one or two residues;

or

R^1.1Is phenyl, wherein two adjacent residues together form a five-or six-membered carbocyclic aromatic or nonaromatic ring which optionally, independently of one another, contains one or two N, S or SO replacing a carbon atom on the ring₂Wherein the ring is optionally substituted by C_1-4-alkyl or ═ O substitution;

R²is selected from C_1-6Alkylene-phenyl, C_1-6-alkylene-naphthyl, and C _1-6-alkylene-thiophenyl; each optionally via a radical selected from C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, O-C_1-6-one, two or three residue substitutions of haloalkyl, halogen;

R³is H, C_1-4-an alkyl group;

R⁴is H, C_1-4-an alkyl group;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or N-C_1-4-an alkyl group;

R¹is selected from

·NHR^1.2、NMeR^1.2；

R^1.2Is selected from

Heteroaryl, optionally via a radical selected from C_1-6Alkyl radical, C_2-6-alkenyl, C_2-6-alkynyl, C_3-6-cycloalkyl, CH₂COO-C_1-6Alkyl, CONR^1.2.1R^1.2.2、COR^1.2.3、COO-C_1-6Alkyl, CONH₂、O-C_1-6Alkyl, halogen, CN, SO₂N(C_1-4-alkyl groups)₂Or optionally substituted by one or two residues selected from C_1-6-heteroaryl substituted with one or two residues of alkyl;

heteroaryl, optionally substituted by a five-or six-membered carbocyclic non-aromatic ring, which independently of one another contains two N, O, S or SO replacing carbon atoms on the ring₂；

Benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each of which is optionally substituted with a group selected from N (C)_1-6-alkyl groups)₂、CONH-C_1-6-alkyl, ═ O substituted with one or two residues;

piperidinyl, optionally substituted with pyridinyl;

4, 5-dihydro-naphtho [2,1-d ]]Thiazoles, optionally via NHCO-C_1-6-an alkyl substitution,

R^1.2.1H、C_1-6alkyl radical, C_1-6alkylene-C _3-6-cycloalkyl, C_1-4Alkylene-phenyl, C_1-4Alkylene-furyl radical, C_3-6-cycloalkyl, C_1-4alkylene-O-C_1-4Alkyl radical, C_1-6Haloalkyl or a five-or six-membered carbocyclic ringNon-aromatic rings optionally containing, independently of one another, one or two N, O, S or SO replacing a carbon atom on the ring₂Optionally substituted with 4-cyclopropylmethyl-piperazinyl;

R^1.2.2is H, C_1-6-an alkyl group;

R^1.2.3is a five-or six-membered carbocyclic non-aromatic ring optionally containing, independently of one another, one or two N, O, S or SO replacing a carbon atom on the ring₂；

R²Is selected from C_1-6Alkylene-phenyl, C_1-6-alkylene-naphthyl, and C_1-6-alkylene-thiophenyl; each of which is optionally selected from C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, O-C_1-6-one, two or three residue substitutions of haloalkyl, halogen;

R³is H, C_1-4-an alkyl group;

R⁴is H, C_1-4-an alkyl group;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1 (above), wherein

A is CH₂O or N-C_1-4-an alkyl group;

R¹is selected from

·NHR^1.2、NMeR^1.2；

R^1.2Is selected from

Heteroaryl, optionally via a radical selected from C_1-6Alkyl radical, C_2-6-alkenyl, C_2-6-alkynyl, C_3-6-cycloalkyl, CH₂COO-C_1-6Alkyl, CONR^1.2.1R^1.2.2、COR^1.2.3、COO-C_1-6Alkyl, CONH₂、O-C_1-6Alkyl, halogen, CN, SO₂N(C_1-4-alkyl groups)₂Or optionally substituted by one or two residues selected from C _1-6-heteroaryl substituted with one or two residues of alkyl;

heteroaryl, optionally via pentaSubstituted by a mono-or six-membered carbocyclic non-aromatic ring, which independently of one another contains two N, O, S or SO replacing carbon atoms on the ring₂；

R^1.2.1Is H, C_1-6Alkyl radical, C_1-6alkylene-C_3-6-cycloalkyl, C_1-4Alkylene-phenyl, C_1-4Alkylene-furyl radical, C_3-6-cycloalkyl, C_1-4alkylene-O-C_1-4Alkyl radical, C_1-6Haloalkyl or a five-or six-membered carbocyclic non-aromatic ring optionally containing, independently of one another, one or two N, O, S or SO replacing carbon atoms on the ring₂Optionally substituted with 4-cyclopropylmethyl-piperazinyl;

R^1.2.2is H, C_1-6-an alkyl group;

R^1.2.3is a five-or six-membered carbocyclic non-aromatic ring, optionally containing, independently of one another, one or two N, O, S or SO replacing carbon atoms on the ring₂；

R²Is selected from C_1-6Alkylene-phenyl, C_1-6-alkylene-naphthyl, and C_1-6-alkylene-thiophenyl; each of which is optionally selected from C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, O-C_1-6-one, two or three residue substitutions of haloalkyl, halogen;

R³is H, C_1-4-an alkyl group;

R⁴is H, C_1-4-an alkyl group;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or N-C _1-4-an alkyl group;

R¹is selected from

·NHCH₂-R^1.3；

R^1.3Selected from phenyl, pyrazolyl, isoAzolyl, pyridyl, pyrimidinyl, indolyl orOxadiazolyl, each of which is optionally selected from C_1-6Alkyl radical, C_3-6-cycloalkyl, O-C_1-6Alkyl, O-C_1-6-one or two residue substitutions of haloalkyl, phenyl, pyrrolidinyl;

R²is selected from C_1-6Alkylene-phenyl, C_1-6-alkylene-naphthyl, and C_1-6-alkylene-thiophenyl; each of which is optionally selected from C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, O-C_1-6-one, two or three residue substitutions of haloalkyl, halogen;

R³is H, C_1-4-an alkyl group;

R⁴is H, C_1-4-an alkyl group;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or N-C_1-4-an alkyl group;

R¹is selected from

·NHR^1.1、NMeR^1.1；

·NHR^1.2、NMeR^1.2；

·NHCH₂-R^1.3；

·NH-C_3-6-cycloalkyl, optionally wherein one carbon atom is replaced by a nitrogen atom, wherein the ring is optionally substituted by a group selected from C_1-6Alkyl, O-C_1-6Alkyl, NHSO₂Phenyl, NHCONH-phenyl, halogen, CN, SO₂-C_1-6-alkyl, COO-C_1-6-one or two residue substitutions of alkyl;

·C_{9 or 10}-bicyclic ring, wherein one or two carbon atoms are replaced by nitrogen atoms and the ring system is bonded to the basic structure of formula 1 via nitrogen atoms and whereinThe ring system is optionally selected from C _1-6-alkyl, COO-C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, NO₂Halogen, CN, NHSO₂-C_1-6-alkyl, m-methoxyphenyl, substituted by one or two residues;

selected from NHCH (pyridyl) CH₂COO-C_1-6Alkyl, NHCH (CH)₂O-C_1-6-alkyl) -benzimidazolyl, optionally substituted with Cl; or

1-aminocyclopentyl, optionally methyl-Substituted by diazole group;

R^1.1is phenyl, optionally via a radical selected from C_1-6Alkyl radical, C_1-6-haloalkyl, CH₂CON(C_1-6-alkyl groups)₂、CH₂NHCONH-C_3-6-cycloalkyl, CN, CONR^1.1.1R^1.1.2、COO-C_1-6Alkyl, O-C_1-6Alkyl, SO₂-C_1-6Alkyl, SO₂-C_1-6alkylene-OH, SO₂-C_3-6-cycloalkyl, SO₂Piperidinyl, SO₂NH-C_1-6Alkyl, SO₂N(C_1-6-alkyl groups)₂Halogen, CN, CO-morpholinyl, CH₂-one or two residues of pyridyl, or optionally substituted by a group selected from C_1-6-alkyl, NHC_1-6-alkyl, ═ O, and heterocycles substituted with one or two residues;

R^1.1.1is H, C_1-6Alkyl radical, C_3-6-cycloalkyl, C_1-6-haloalkyl, CH₂CON(C_1-6-alkyl groups)₂、CH₂CO-Azacyclobutyl, C_1-6alkylene-C_3-6-cycloalkyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, C_1-6alkylene-OH or thiadiazolyl, optionally via C_1-6-alkyl substitution;

R^1.1.2is H, C_1-6Alkyl, SO₂C_1-6-alkanesA group;

or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one O replacing a carbon atom on the ring, optionally via a substituent selected from CH ₂One or two residue substitutions of OH;

R^1.2is selected from

Heteroaryl, optionally via a radical selected from C_1-6Alkyl radical, C_3-6-cycloalkyl, CH₂COO-C_1-6Alkyl, CONR^1.2.1R^1.2.2、COO-C_1-6Alkyl, CONH₂、O-C_1-6-alkyl, halogen, CN, CO-pyrrolidinyl, CO-morpholinyl, or optionally substituted with one or two residues selected from C_1-6-heteroaryl substituted with one or two residues of alkyl;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each of which is optionally substituted with a group selected from N (C)_1-6-alkyl groups)₂、CONH-C_1-6-alkyl, ═ O substituted with one or two residues;

piperidinyl, optionally substituted with pyridinyl;

4, 5-dihydro-naphtho [2,1-d ]]Thiazoles, optionally via NHCO-C_1-6-an alkyl substitution,

R^1.2.1is H, C_1-6-an alkyl group;

R^1.2.2is H, C_1-6-an alkyl group;

R^1.3selected from phenyl, pyrazolyl, isoAzolyl, pyrimidinyl, indolyl orOxadiazolyl, each of which is optionally selected from C_1-6Alkyl radical, C_3-6-cycloalkyl, O-C_1-6Alkyl, O-C_1-6-one or two residue substitutions of haloalkyl;

R²is selected from C_1-6Alkylene-phenyl or C_1-6-alkylene-naphthyl, both optionally selected from C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6Alkyl, O-C_1-6-haloalkyl, halogen substituted with one or two residues; or CH₂-thiophenyl, optionally substituted with one or two residues selected from halogen;

R³Is H, C_1-4-an alkyl group;

R⁴is H, C_1-4-an alkyl group;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

·NHR^1.2、NMeR^1.2；

·NHCH₂-R^1.3；

NH-cyclohexyl, optionally via a radical selected from C_1-4Alkyl, NHSO₂-phenyl, NHCONH-phenyl, halo substituted with one or two residues;

NH-pyrrolidinyl, optionally via a substituent selected from SO₂-C_1-4-alkyl, COO-C_1-4-one or two residue substitutions of alkyl;

piperidinyl, optionally via a substituent selected from NHSO₂-C_1-4-alkyl, m-methoxyphenyl, substituted by one or two residues;

dihydro-indolyl, dihydro-isoindolyl, tetrahydro-quinolyl or tetrahydro-isoquinolyl, optionally via a substituent selected from C_1-4-alkyl, COO-C_1-4Alkyl radical, C_1-4Haloalkyl, O-C_1-4Alkyl, NO₂One or two residue substitutions of halogen;

selected from NHCH (pyridyl) CH₂COO-C_1-4Alkyl, NHCH (CH)₂O-C_1-4-alkyl) -benzimidazolyl, optionally substituted with Cl; or

1-aminocyclopentyl, optionally methyl-Substituted by diazole group;

R^1.1is phenyl, optionally via a radical selected from C_1-4Alkyl radical, C_1-4-haloalkyl, CH₂CON(C_1-4-alkyl groups)₂、CH₂NHCONH-C_3-6-cycloalkyl, CN, CONR^1.1.1R^1.1.2、COO-C_1-4Alkyl, O-C_1-4Alkyl, SO₂-C_1-4Alkyl, SO₂-C_1-4alkylene-OH, SO₂-C_3-6-cycloalkyl, SO₂Piperidinyl, SO ₂NH-C_1-4Alkyl, SO₂N(C_1-4-alkyl groups)₂Halogen, CO-morpholinyl, CH₂-pyridyl substituted by one or two residues, or imidazolidinyl, piperidinyl, oxazaheterocycloalkyl, pyrazolyl, triazolyl, tetrazolyl,Azolyl group,Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each optionally selected from C_1-4-alkyl, NHC_1-4-alkyl, ═ O substituted with one or two residues;

R^1.1.1is H, C_1-6Alkyl radical, C_3-6-cycloalkyl, C_1-4Haloalkyl, CH₂CON(C_1-4-alkyl groups)₂、CH₂CO-Azacyclobutyl, C_1-4alkylene-C_3-6-cycloalkyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, C_1-4alkylene-OH or thiadiazolyl, optionally via C_1-4-alkyl substitution;

R^1.1.2is H, C_1-4Alkyl, SO₂C_1-4-an alkyl group;

or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containingReplacing one O of the ring carbon atoms, optionally via a group selected from CH₂One or two residue substitutions of OH;

R^1.2is selected from

Pyridinyl, pyridazinyl, pyrrolyl, pyrazolyl, isoOxazolyl, thiazolyl, thiadiazolyl, optionally via C_1-4Alkyl radical, C_3-6-cycloalkyl, CH₂COO-C_1-4Alkyl, CONR^1.2.1R^1.2.2、COO-C_1-4Alkyl, CONH₂、O-C_1-4-alkyl, halogen, CO-pyrrolidinyl, CO-morpholinyl, substituted by one or two residues, or pyrazolyl, triazolyl, tetrazolyl, isoxazolyl Azolyl group,Oxadiazolyl, each of which is optionally selected from C_1-4-one or two residue substitutions of alkyl;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each of which is optionally substituted with a group selected from N (C)_1-4-alkyl groups)₂、CONH-C_1-4-alkyl, ═ O substituted with one or two residues;

piperidinyl, optionally substituted with pyridinyl;

4, 5-dihydro-naphtho [2,1-d ]]Thiazoles, optionally via NHCO-C_1-4-an alkyl substitution,

R^1.2.1is H, C_1-4-an alkyl group;

R^1.2.2is H, C_1-4-an alkyl group;

R^1.3selected from phenyl, pyrazolyl, isoAzolyl, pyrimidinyl, indolyl orOxadiazolyl, each of which is optionally selected from C_1-4Alkyl radical, C_3-6-cycloalkyl, O-C_1-4Alkyl, O-C_1-4-one or two residue substitutions of haloalkyl;

R²is selected from C_1-6Alkylene-phenyl or C_1-6-alkylene-naphthyl, both optionally selected from C_1-4Alkyl radical, C_1-4Haloalkyl, O-C_1-4-haloalkyl, halogen, substitution of one or two residues; or CH₂-thiophenyl, optionally substituted by one or two residues selected from halogen;

R³is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

·NHR^1.2、NMeR^1.2；

·NHCH₂-R^1.3；

NH-piperidinyl, optionally substituted with pyridinyl;

NH-cyclohexyl, optionally via a radical selected from t-Bu, NHSO ₂-phenyl, NHCONH-phenyl, substitution of one or two residues of F;

NH-pyrrolidinyl, optionally via a radical selected from SO₂One or two residues of Me and COO-t-Bu;

piperidinyl, optionally via a radical selected from NHSO₂-n-Bu, one or two residue substitutions of m-methoxyphenyl;

dihydro-indolyl, dihydro-isoindolyl, tetrahydro-quinolyl or tetrahydro-isoquinolyl, optionally via a substituent selected from Me, COOMe, CF₃、OMe、NO₂F, Br;

selected from NHCH (pyridyl) CH₂COOMe、NHCH(CH₂OMe) -benzimidazolyl, optionally substituted with Cl; or

1-aminocyclopentyl, optionally methyl-Substituted by diazole group;

R^1.1is phenyl, optionally via a catalyst selected from Me, Et, t-Bu, CF₃、CH₂CONMe₂、CH₂NHCONH-cyclohexyl, CN, CONR^1.1.1R^1.1.2、COOMe、COOEt、OMe、SO₂Me、SO₂CH₂CH₂OH、SO₂Et、SO₂-cyclopropyl, SO₂Piperidinyl, SO₂NHEt、SO₂NMeEt, F, Cl, CO-morpholinyl, CH₂-pyridyl substituted by one or two residues, or imidazolidinyl, piperidinyl, oxazaheterocycloalkyl, pyrazolyl, triazolyl, tetrazolyl, triazolyl, and triazolyl,Azolyl group,Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each optionally substituted with one or two residues selected from Me, NHMe, ═ O;

R^1.1.1is H, Me, Et, t-Bu, i-Pr, cyclopropyl, CH₂-i-Pr、CH₂-t-Bu、CH(CH₃)CH₂CH₃、CH₂CHF₂、CH₂CONMe₂、CH₂CO-AZACYCLOBUTYL, CH₂-cyclopropyl, CH₂-cyclobutyl, CH₂Pyranyl, CH ₂-tetrahydrofuranyl, CH₂-furyl, CH₂CH₂OH or thiadiazolyl, optionally substituted with Me;

R^1.1.2is H, Me, Et, SO₂Me、SO₂Et；

Or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing substitution of carbon atoms on the ringOptionally via a group selected from CH₂One or two residue substitutions of OH;

R^1.2is selected from

Pyridinyl, pyrrolyl, pyrazolyl, isoOxazolyl, thiazolyl, thiadiazolyl, optionally substituted by a group selected from Me, Et, Pr, Bu, cyclopropyl, CH₂COOEt、CONR^1.2.1R^1.2.2、COOMe、COOEt、CONH₂One or two residues of OMe, Cl, Br, CO-pyrrolidinyl, CO-morpholinyl, or pyrazolyl, triazolyl, tetrazolyl, isoxazolylAzolyl group,(ii) oxadiazolyl, each optionally substituted with Me;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with NMe₂One or two residue substitutions of CONHMe, ═ O;

4, 5-dihydro-naphtho [2,1-d ] thiazole, optionally substituted with NHCOMe,

R^1.2.1h, Me;

R^1.2.2h, Me;

R¹3 is selected from phenyl, pyrazolyl, isoAzolyl, pyrimidinyl, indolyl orOxadiazolyl, each optionally substituted with one or more substituents selected from the group consisting of Me, Et, Pr, cyclopentyl, OMe, OCHF₂One or two residue substitutions;

R²is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via CH₃、CF₃、OCF₃、F、Cl、Br、EtOne or two residue substitutions; or CH ₂-thiophenyl, optionally substituted with one or two residues selected from Cl, Br;

R³is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1

·NHR^1.2，

R^1.1Is phenyl, optionally via a catalyst selected from Me, Et, Pr, Bu, CF₃、CH₂CONMe₂、CH₂NHCONH-cyclohexyl, CN, CONR^1.1.1R^1.1.2、COOMe、COOEt、OMe、SO₂Me、SO₂CH₂CH₂OH、SO₂Et、SO₂-cyclopropyl, SO₂Piperidinyl, SO₂NHEt、SO₂NMeEt, F, Cl, CO-morpholinyl, CH₂-pyridyl substituted by one or two residues, or imidazolidinyl, piperidinyl, oxazaheterocycloalkyl, pyrazolyl, triazolyl, tetrazolyl, triazolyl, and triazolyl,Azolyl group,Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each optionally substituted with one or two residues selected from Me, NHMe, ═ O;

R^1.1.1is H, Me, Et, t-Bu, i-Pr, cyclopropyl, CH₂-i-Pr、CH₂-t-Bu、CH(CH₃)CH₂CH₃、CH₂CHF₂、CH₂CONMe₂、CH₂CO-AZACYCLOBUTYL, CH₂-cyclopropyl, CH₂-cyclobutyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, CH₂CH₂OH or thiadiazolyl, optionally substituted with Me;

R^1.1.2is H, Me, Et, SO₂Me、SO₂Et；

Or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one O replacing a carbon atom on the ring, optionally via a substituent selected from CH₂One or two residue substitutions of OH;

R^1.2is selected from

Pyridinyl, pyrrolyl, pyrazolyl, iso Oxazolyl, thiazolyl, thiadiazolyl, optionally substituted by a group selected from Me, Et, Pr, Bu, cyclopropyl, CH₂COOEt、CONR^1.2.1R^1.2.2、COOMe、COOEt、CONH₂One or two residues of OMe, Cl, Br, CO-pyrrolidinyl, CO-morpholinyl, or pyrazolyl, triazolyl, tetrazolyl, isoxazolylAzolyl group,(ii) oxadiazolyl, each optionally substituted with Me;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with NMe₂One or two residue substitutions of CONHMe, ═ O;

4, 5-dihydro-naphtho [2,1-d ] thiazole, optionally substituted with NHCOMe,

R^1.2.1h, Me;

R^1.2.2h, Me;

R²is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via CH₃、CF₃、OCF₃One or two residues of F, Cl, Br, Et;

R³is H;

R⁴is H.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

·NHR^1.2、NMeR^1.2；

·NHCH₂-R^1.3；

R^1.1Is phenyl, optionally via a catalyst selected from Me, Et, Pr, Bu, CF₃、CH₂CONMe₂、CH₂NHCONH-cyclohexyl, CN, CONR^1.1.1R^1.1.2、COOMe、COOEt、OMe、SO₂Me、SO₂CH₂CH₂OH、SO₂Et、SO₂-cyclopropyl, SO₂Piperidinyl, SO₂NHEt、SO₂NMeEt, F, Cl, CO-morpholinyl, CH₂-pyridyl substituted by one or two residues, or imidazolidinyl, piperidinyl, oxazaheterocycloalkyl, pyrazolyl, triazolyl, tetrazolyl,Azolyl group,Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each optionally substituted with one or two residues selected from Me, NHMe, ═ O;

R^1.1.1Is H, Me, Et, Pr, Bu, cyclopropyl, CH₂-Pr、CH₂-Bu、CH(CH₃)CH₂CH₃、CH₂CHF₂、CH₂CONMe₂、CH₂CO-AZACYCLOBUTYL, CH₂-cyclopropyl, CH₂-cyclobutyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, CH₂CH₂OH or thiadiazolyl, optionally substituted with Me;

R^1.1.2h, Me is,Et、SO₂Me、SO₂Et；

Or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one O replacing a carbon atom on the ring, optionally via a substituent selected from CH₂One or two residue substitutions of OH;

R^1.2is selected from

Pyridinyl, pyrrolyl, pyrazolyl, isoOxazolyl, thiazolyl, thiadiazolyl, optionally substituted by a group selected from Me, Et, Pr, Bu, cyclopropyl, CH₂COOEt、CONR^1.2.1R^1.2.2、COOMe、COOEt、CONH₂One or two residues of OMe, Cl, Br, CO-pyrrolidinyl, CO-morpholinyl, or pyrazolyl, triazolyl, tetrazolyl, isoxazolylAzolyl group,(ii) oxadiazolyl, each optionally substituted with Me;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with NMe₂One or two residue substitutions of CONHMe, ═ O;

4, 5-dihydro-naphtho [2,1-d ] thiazole, optionally substituted with NHCOMe,

R^1.2.1h, Me;

R^1.2.2h, Me;

R^1.3selected from phenyl, pyrazolyl, isoAzolyl, pyrimidinyl, indolyl orOxadiazolyl, each optionally substituted with one or more substituents selected from the group consisting of Me, Et, Pr, cyclopentyl, OMe, OCHF ₂By substitution of one or two residues；

R²Is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via CH₃、CF₃、OCF₃One or two residues of F, Cl, Br, Et; or CH₂-thiophenyl, optionally substituted with one or two residues selected from Cl, Br;

R³is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

R^1.1Is phenyl, optionally via a catalyst selected from Me, Et, t-Bu, CF₃、CH₂CONMe₂、CH₂NHCONH-cyclohexyl, CN, CONR^1.1.1R^1.1.2、COOMe、COOEt、OMe、SO₂Me、SO₂CH₂CH₂OH、SO₂Et、SO₂-cyclopropyl, SO₂Piperidinyl, SO₂NHEt、SO₂NMeEt, F, Cl, CO-morpholinyl, CH₂-pyridyl substituted by one or two residues, or imidazolidinyl, piperidinyl, oxazaheterocycloalkyl, pyrazolyl, triazolyl, tetrazolyl,Azolyl group,Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each optionally substituted with one or two residues selected from Me, NHMe, ═ O;

R^1.1.1is H, Me, Et, Bu, Pr, cyclopropyl, CH₂-Pr、CH₂-Bu、CH(CH₃)CH₂CH₃、CH₂CHF₂、CH₂CONMe₂、CH₂CO-AZACYCLOBUTYL, CH₂-cyclopropyl, CH₂-cyclobutyl, CH₂-pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, CH₂CH₂OH or thiadiazolyl, optionally substituted with Me;

R^1.1.2is H, Me, Et, SO₂Me、SO₂Et

Or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one O replacing a carbon atom on the ring, optionally via a substituent selected from CH ₂One or two residue substitutions of OH;

R²is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via CH₃、CF₃、OCF₃One or two residues of F, Cl, Br, Et; or CH₂-thiophenyl, optionally substituted with one or two residues selected from Cl, Br;

R³is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

R^1.1Is phenyl, optionally via a catalyst selected from Me, Et, t-Bu, CF₃、CH₂CONMe₂、CH₂NHCONH-cyclohexyl, CN, CONR^1.1.1R^1.1.2、COOMe、COOEt、OMe、SO₂Me、SO₂CH₂CH₂OH、SO₂Et、SO₂-cyclopropyl, SO₂Piperidinyl, SO₂NHEt、SO₂NMeEt, F, Cl, CO-morpholinyl, CH₂-substitution of one or two residues of the pyridyl group, or of the imidazopyridinyl, piperidinyl, oxazacylyl, pyrazolyl, triazolyl, tetra-aminoAzolyl group,Azolyl group,Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each optionally substituted with one or two residues selected from Me, NHMe, ═ O;

R^1.1.1is H, Me, Et, Bu, Pr, cyclopropyl, CH₂-Pr、CH₂-Bu、CH(CH₃)CH₂CH₃、CH₂CHF₂、CH₂CONMe₂、CH₂CO-AZACYCLOBUTYL, CH₂-cyclopropyl, CH₂-cyclobutyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, CH₂CH₂OH or thiadiazolyl, optionally substituted with Me;

R^1.1.2is H, Me, Et, SO₂Me、SO₂Et

Or R^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one O replacing a carbon atom on the ring, optionally via a substituent selected from CH ₂One or two residue substitutions of OH;

R²as defined in table 1 shown below;

R³is H;

R⁴is H.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

R^1.1Is phenyl, optionally via a catalyst selected from Me, Et, t-Bu, CF₃、CH₂CONMe₂、CH₂NHCONH-cyclohexyl, CN, CONR^1.1.1R^1.1.2、COOMe、COOEt、OMe、SO₂Me、SO₂CH₂CH₂OH、SO₂Et、SO₂-cyclopropyl, SO₂Piperidinyl, SO₂NHEt、SO₂NMeEt, F, Cl, CO-morpholinyl, CH₂-substitution of one or two residues of the pyridyl group, or of the imidazopyridinyl, piperidinyl, oxazacylyl, pyrazolyl, triazolyl, tetrazolyl groups,Azolyl group,Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each optionally substituted with one or two residues selected from Me, NHMe, ═ O;

and R is^1.1.1And R^1.1.2Together form a four-, five-or six-membered carbocyclic ring, optionally containing one O replacing a carbon atom on the ring, optionally via a substituent selected from CH₂One or two residue substitutions of OH;

R²as defined in table 1 shown below;

R³is H;

R⁴is H.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

R^1.1Is phenyl, optionally via a catalyst selected from Me, Et, t-Bu, CF₃、CH₂CONMe₂、CH₂NHCONH-cyclohexyl, CN, CONR^1.1.1R^1.1.2COOMe, COOEt, OMe, F, Cl;

R^1.1.1Is H, Me, Et, Bu, Pr, cyclopropyl, CH₂-Pr、CH₂-Bu、CH(CH₃)CH₂CH₃、CH₂CHF₂、CH₂CONMe₂、CH₂CO-AZACYCLOBUTYL, CH₂-cyclopropyl, CH₂-cyclobutyl, CH₂-pyranyl radical、CH₂-tetrahydrofuranyl, CH₂-furyl, CH₂CH₂OH or thiadiazolyl, optionally substituted with Me;

R^1.1.2is H, Me, Et, SO₂Me、SO₂Et

R²As defined in table 1 shown below;

R³is H;

R⁴is H.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

R^1.1Is phenyl, optionally via a substituent selected from SO₂Me、SO₂CH₂CH₂OH、SO₂Et、SO₂-cyclopropyl, SO₂Piperidinyl, SO₂NHEt、SO₂One or two residue substitutions of NMeEt;

R^1.1.1is H, Me, Et, Bu, Pr, cyclopropyl, CH₂-Pr、CH₂-Bu、CH(CH₃)CH₂CH₃、CH₂CHF₂、CH₂CONMe₂、CH₂CO-AZACYCLOBUTYL, CH₂-cyclopropyl, CH₂-cyclobutyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, CH₂CH₂OH or thiadiazolyl, optionally substituted with Me;

R^1.1.2is H, Me, Et, SO₂Me、SO₂Et；

R²As defined in table 1 shown below;

R³is H;

R⁴is H.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.1、NMeR^1.1；

R^1.1Is phenyl, optionally via a catalyst selected from Me, Et, t-Bu, CF₃、CH₂CONMe₂、CH₂NHCONH-cyclohexyl, CN, CONR^1.1.1R^1.1.2、COOMe、COOEt、OMe、SO₂Me、SO₂CH₂CH₂OH、SO₂Et、SO₂-cyclopropyl, SO₂Piperidinyl, SO₂NHEt、SO₂One residue of NMeEt, F, Cl and additionally via a residue selected from CO-morpholinyl, CH₂-one residue of a pyridyl group, or an imidazopyridinyl, piperidinyl, oxazacylohexyl, pyrazolyl, triazolyl, tetrazolyl, triazolyl, and triazolyl group, Azolyl group,Oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl, each optionally substituted with one or two residues selected from Me, NHMe, ═ O;

R^1.1.1is H, Me, Et, Bu, Pr, cyclopropyl, CH₂-Pr、CH₂-Bu、CH(CH₃)CH₂CH₃、CH₂CHF₂、CH₂CONMe₂、CH₂CO-AZACYCLOBUTYL, CH₂-cyclopropyl, CH₂-cyclobutyl, CH₂Pyranyl, CH₂-tetrahydrofuranyl, CH₂-furyl, CH₂CH₂OH or thiadiazolyl, optionally substituted with Me;

R^1.1.2is H, Me, Et, SO₂Me、SO₂Et

R²As defined in table 1 shown below;

R³is H;

R⁴is H.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.2、NMeR^1.2；

R^1.2Is selected from

Pyridinyl, pyridazinyl, pyrrolyl, pyrazolyl, isoOxazolyl, thiazolyl, thiadiazolyl, optionally substituted by a group selected from Me, Et, Pr, Bu, cyclopropyl, CH₂COOEt、CONR^1.2.1R^1.2.2、COOMe、COOEt、CONH₂One or two residues of OMe, Cl, Br, CO-pyrrolidinyl, CO-morpholinyl, or pyrazolyl, triazolyl, tetrazolyl, isoxazolylAzolyl group,(ii) oxadiazolyl, each optionally substituted with Me;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with NMe₂One or two residue substitutions of CONHMe, ═ O;

4, 5-dihydro-naphtho [2,1-d ] thiazole, optionally substituted with NHCOMe,

R^1.2.1h, Me;

R^1.2.2H, Me;

R²is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via CH₃、CF₃、OCF₃One or two residues of F, Cl, Br, Et; or CH₂-thiophenyl, optionally substituted with one or two residues selected from Cl, Br;

R³is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

·NHR^1.2、NMeR^1.2；

R^1.2Selected from pyridyl, pyridazinyl, pyrrolyl, pyrazolyl, isopyrazolylOxazolyl, thiazolyl, thiadiazolyl, optionally substituted with one or more substituents selected from Me, Et, n-Pr, i-Pr, Bu, cyclopropyl, CH₂COOEt、CONR^1.2.1R^1.2.2、COOMe、COOEt、CONH₂One or two residues of OMe, Cl, Br, CO-pyrrolidinyl, CO-morpholinyl, or pyrazolyl, triazolyl, tetrazolyl, isoxazolylAzolyl group,(ii) oxadiazolyl, each optionally substituted with Me;

R^1.2.1h, Me;

R^1.2.2h, Me;

R²is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via CH₃、CF₃、OCF₃One or two residues of F, Cl, Br, Et; or CH₂-thiophenyl, optionally substituted with one or two residues selected from Cl, Br;

R³is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹Is selected from

·NHCH₂-R^1.3；

R^1.3Selected from phenyl, pyrazolyl, isoAzolyl, pyrimidinyl, indolyl orOxadiazolyl, each optionally substituted with one or more substituents selected from the group consisting of Me, Et, Pr, cyclopentyl, OMe, OCHF₂One or two residue substitutions;

R²is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via CH₃、CF₃、OCF₃One or two residues of F, Cl, Br, Et; or CH₂-thiophenyl, optionally substituted with one or two residues selected from Cl, Br;

R³is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe;

R¹is selected from

NH-piperidinyl, optionally substituted with pyridinyl;

NH-cyclohexyl, optionally via a gas selected from t-Bu, NHSO₂-phenyl, NHCONH-phenyl, substitution of one or two residues of F;

NH-pyrrolidinyl, optionally via a radical selected from SO₂One or two residues of Me and COO-t-Bu;

piperidinyl, optionally via a radical selected from NHSO₂-n-Bu, one or two residue substitutions of m-methoxyphenyl;

dihydro-indolyl, dihydro-isoindolyl, tetrahydro-quinolyl or tetrahydro-isoquinolyl, optionally via a substituent selected from Me, COOMe, CF₃、OMe、NO₂F, Br;

selected from NHCH (pyridyl) CH₂COOMe、NHCH(CH₂OMe) -benzimidazolyl, optionally substituted with Cl; or

1-aminocyclopentyl, optionally methyl-Substituted by diazole group;

R²is selected from CH₂-phenyl or CH₂-naphthyl, both optionally via CH₃、CF₃、OCF₃One or two residues of F, Cl, Br, Et; or CH₂-thiophenyl, optionally substituted with one or two residues selected from Cl, Br;

R³is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the invention also includes administering to a subject a compound of formula 1, wherein A is CH₂O or NMe, R¹Selected from NHR^1.2、NMeR^1.2；R²As defined in table 1 shown below; r³Is H; r⁴Is H and R^1.2Is selected from

Pyridyl, optionally via a catalyst selected from Me, Et, i-Pr, n-Bu, cyclopropyl, CONR^1.2.1R^1.2.2、COOMe、COOEt、CONH₂One or two residues of OMe, Cl, Br, CO-pyrrolidinyl, CO-morpholinyl, or pyrazolyl, triazolyl, tetrazolyl, isoxazolylAzolyl group,(ii) oxadiazolyl, each optionally substituted with Me;

pyrrolyl, optionally substituted by one or two residues selected from Me, Et, COOMe, COOEt;

pyrazolyl, optionally substituted by one or two residues selected from Me, Et, cyclopropyl, COOEt, CO-pyrrolidinyl;

different from one another(ii) oxazolyl, optionally substituted with one or two residues selected from t-Bu, COOEt;

thiazolyl, optionally selected from Me, n-Pr, i-Pr, Bu, COOMe, COOEt, CH ₂COOEt、CONR^1.2.1R^1.2.2One or two residue substitutions;

thiadiazolyl, optionally substituted by one or two residues selected from COOEt;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with NMe₂One or two residue substitutions of CONHMe, ═ O;

4, 5-dihydro-naphtho [2,1-d ] thiazole, optionally substituted with NHCOMe,

and is

R^1.2.1Is H or Me;

R^1.2.2is H or Me.

Another embodiment of the invention also includes administering to a subject a compound of formula 1, wherein A is CH₂O or NMe, R¹Selected from NHR^1.2、NMeR^1.2；R²As defined in table 1 shown below; r³Is H; r⁴Is H and R^1.2Is selected from

Pyridyl, optionally via a catalyst selected from Me, Et, i-Pr, n-Bu, CONR^1.2.1R^1.2.2、COOMe、COOEt、CONH₂One or two residues of OMe, Cl, Br;

pyrrolyl, optionally substituted by one or two residues selected from Me, Et, COOMe, COOEt;

pyrazolyl, optionally substituted by one or two residues selected from Me, Et, cyclopropyl, COOEt, CO-pyrrolidinyl;

different from one another(ii) oxazolyl, optionally substituted with one or two residues selected from t-Bu, COOEt;

thiazolyl, optionally selected from Me, n-Pr, i-Pr, Bu, COOMe, COOEt, CONR^1.2.1R^1.2.2One or two residue substitutions;

thiadiazolyl, optionally substituted by one or two residues selected from COOEt;

Benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with NMe₂One or two residue substitutions of CONHMe, ═ O;

and is

R^1.2.1Is H or Me;

R^1.2.2is H or Me.

Another embodiment of the present invention further comprises administering to the subject a compound of formula 1, wherein

A is CH₂O or NMe, R¹Selected from NHR^1.2、NMeR^1.2；R²As defined in table 1 shown below; r³Is H; r⁴Is H; r^1.2Is pyridyl, optionally via a catalyst selected from Me, Et, i-Pr, n-Bu, CONR^1.2.1R^1.2.2、COOMe、COOEt、CONH₂One or two residues of OMe, Cl, Br; r^1.2.1Is H or Me and R^1.2.2Is H or Me.

A is CH₂O or NMe, R¹Selected from NHR^1.2、NMeR^1.2；R²As defined in table 1 shown below; r³Is H; r⁴Is H; r^1.2Is pyrrolyl, optionally substituted with one or two residues selected from Me, Et, COOMe, COOEt; r^1.2.1Is H or Me and R^1.2.2Is H or Me.

A is CH₂O or NMe, R¹Selected from NHR^1.2、NMeR^1.2；R²As defined in table 1 shown below; r³Is H; r⁴Is H; r^1.2Is pyrazolyl, optionally substituted with one or two residues selected from Me, Et, cyclopropyl, COOEt, CO-pyrrolidinyl; r^1.2.1Is H or Me and R^1.2.2Is H or Me.

A is CH₂O or NMe, R¹Selected from NHR^1.2、NMeR^1.2；R²As defined in table 1 shown below; r³Is H; r⁴Is H; r^1.2Is different from(ii) oxazolyl, optionally substituted with one or two residues selected from t-Bu, COOE; r ^1.2.1Is H or Me and R^1.2.2Is H or Me.

A is CH₂O or NMe, R¹Selected from NHR^1.2、NMeR^1.2；R²As defined in table 1 shown below; r³Is H; r⁴Is H; r^1.2Thiazolyl, optionally selected from Me, n-Pr, i-Pr, Bu, COOMe, COOEt, CONR^1.2.1R^1.2.2One or two residue substitutions; r^1.2.1Is H or Me and R^1.2.2Is H or Me.

A is CH₂O or NMe, R¹Selected from NHR^1.2、NMeR^1.2；R²As defined in table 1 shown below; r³Is H; r⁴Is H; r^1.2Is thiadiazolyl, optionally substituted by one or two residues selected from COOEt; r^1.2.1Is H or Me and R^1.2.2Is H or Me.

A is CH₂O or NMe, R¹Selected from NHR^1.2、NMeR^1.2；R²As defined in table 1 shown below; r³Is H; r⁴Is H; r^1.2Is benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolyl, each of which is optionally substituted with NMe₂One or two residue substitutions of CONHMe, ═ O; r^1.2.1Is H or Me and R^1.2.2Is H or Me.

Another embodiment of the present invention also includes administering to a subject a compound of formula 1, wherein all radicals are as defined above, except that R is^1.3Is selected from

Phenyl, optionally via OCHF₂Substitution;

pyrazolyl, optionally substituted by Me or Et;

different from one another(iv) oxazolyl, optionally substituted with Pr;

pyrimidinyl, optionally substituted with two OMe;

an indolyl group;

·(ii) oxadiazolyl, optionally substituted with cyclopentyl.

Another embodiment of the invention also includes administering to a subject a compound of formula 1, wherein all groups are as defined above, except that A is CH₂。

Another embodiment of the present invention also includes administering to a subject a compound of formula 1, wherein all groups are as defined above except a is O.

Another embodiment of the invention also includes administering to a subject a compound of formula 1, wherein all groups are as defined above except a is NMe.

Another embodiment of the present invention are compounds of formula 1 wherein

A is CH₂O or NMe;

R¹is selected from

R²Is selected from

R³Is H;

R⁴is H;

or R³And R⁴Together form CH₂-CH₂A group.

Another embodiment of the invention are compounds of formula 1, wherein a is as defined above; r³Is H; r⁴Is H; and R²As defined in table 1 shown below; and R¹Is selected from

Another embodiment of the present invention also includes administering to a subject a compound of formula 1, wherein a is as defined above; r³Is H; r⁴Is H; and R²As defined in table 1 shown below; and R¹Is selected from

Another embodiment of the invention also includes administering to a subject a compound of formula 1, wherein a is as defined above; r³Is H; r⁴Is H; and R²As defined in table 1 shown below; r¹Is selected from

Another embodiment of the present invention also includes administering to a subject a compound of formula 1, wherein a is as defined above; r ³Is H; r⁴Is H; and R²As defined in table 1 shown below; and R¹Is selected from

Another embodiment of the present invention also includes administering to a subject a compound of formula 1, wherein a is as defined above; r³Is H; r⁴Is H; and R²As defined in table 1 shown below; r¹Is selected from

Table 1: r²Defined as one of the groups shown in definitions 1 to 4 below:

another embodiment of the present invention also includes administering to a subject a compound of formula 1, wherein the compound of formula 1 is present as an individual optical isomer, a mixture of individual enantiomers, or a racemate, e.g., as an enantiomerically pure compound.

Another embodiment of the present invention also includes administering to a subject a compound of formula 1, wherein the compound of formula 1 is present in the form of its acid addition salt with a pharmaceutically acceptable acid and optionally in the form of a solvate and/or hydrate.

b. Co-crystals and salts

Other embodiments of the invention also include administering to the subject a co-crystal of a compound of formula 2 (below). In general, for groups containing two or more subgroups in this "co-crystal and salt" moiety, the first named subgroup is the point of attachment of the group, e.g., substituent "C _1-3By-alkyl-aryl "is meant an aryl group bonded to a C1-3-alkyl group, the former bonded to the core or group to which the substituent is attached.

Wherein

R¹Is C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6-haloalkyl, halogen;

m is 1, 2 or 3; and in some examples 1 or 2;

R^2aand R^2bEach independently selected from H, C_1-6Alkyl radical, C_1-6-alkenyl, C_1-6-alkynyl, C_3-6-cycloalkyl, COO-C_1-6Alkyl, O-C_1-6Alkyl, CONR^2b.1R^2b.2A halogen;

R^2b.1is H, C_1-6Alkyl radical, C_0-4-alkyl-C_3-6-cycloalkyl, C_1-6-a haloalkyl group;

R^2b.2is H, C_1-6-an alkyl group;

or R^2b.1And R^2b.2Together are C_3-6-alkylene, which forms a heterocyclic ring with the nitrogen atom, wherein optionally one carbon atom of the ring is replaced by an oxygen atom;

R³is H, C_1-6-an alkyl group;

x is an anion selected from chloride, bromide, iodide, sulfate, phosphate, methanesulfonate, nitrate, maleate, acetate, benzoate, citrate, salicylate, fumarate, tartrate, dibenzoyltartrate, oxalate, succinate, benzoate, and p-toluenesulfonate; and in some examples, chloride or dibenzoyl tartrate;

j is 0, 0.5, 1, 1.5 or 2; and in some examples 1 or 2;

wherein the eutectic forming agent is selected from the group consisting of: orotic acid, hippuric acid, L-pyroglutamic acid, D-pyroglutamic acid, nicotinic acid, L- (+) -ascorbic acid, saccharin, piperazine, 3-hydroxy-2-naphthoic acid, mucic acid (galactaric acid), pamoic acid (enbo' ic acid), stearic acid, cholic acid, deoxycholic acid, nicotinamide, isonicotinamide, succinamide, uracil, L-lysine, L-proline, D-valine, L-arginine, glycine, in some examples ascorbic acid, mucic acid, pamoic acid, succinamide, nicotinic acid, nicotinamide, isonicotinamide, L-lysine, L-proline.

Another aspect of the invention also includes administering to a subject a co-crystal of a compound of formula 2, wherein

R^2aIs H, C_1-6Alkyl radical, C_1-6-alkenyl, C_1-6-alkynyl, C_3-6-cycloalkyl, O-C_1-6Alkyl, CONR^2a.1R^2a.2；

R^2a.1Is H, C_1-6Alkyl radical, C_1-6-a haloalkyl group;

R^2a.2is H, C_1-6-an alkyl group;

R^2bis H, C_1-6Alkyl radical, C_1-6-alkenyl, C_1-6-alkynyl, C_3-6-cycloalkyl, COO-C_1-6Alkyl, O-C_1-6Alkyl, CONR^2b.1R^2b.2A halogen;

R^2b.1is H, C_1-6Alkyl radical, C_0-4-alkyl-C_3-6-cycloalkyl, C_1-6-a haloalkyl group;

R^2b.2is H, C_1-6-an alkyl group;

or R^2b.1And R^2b.2Together are C_3-6-alkylene groups forming a heterocyclic ring with the nitrogen atom, wherein optionally one carbon atom of the ring is replaced by an oxygen atom and the remaining residues are as defined above.

Another aspect of the invention also includes administering to a subject a co-crystal of a compound of formula 2, wherein

R^2aIs H, C_1-6Alkyl radical, C_1-6-alkynyl, C_3-6-cycloalkyl, O-C_1-6Alkyl, CONR^2a.1R^2a.2；

R^2a.1Is C_1-6-an alkyl group;

R^2a.2is H;

R^2bis H, C_1-6Alkyl, O-C_1-6Alkyl, CONR^2b.1R^2b.2；

R^2b.1Is C_1-6Alkyl radical, C_0-4-alkyl-C_3-6-cycloalkyl, C_1-6-a haloalkyl group;

R^2b.2is H, C_1-6-an alkyl group;

or R^2b.1And R^2b.2Together are C_3-6-alkylene groups forming a heterocyclic ring with the nitrogen atom, wherein optionally one carbon atom of the ring is replaced by an oxygen atom and the remaining residues are as defined above.

Another aspect of the invention also includes administering to a subject a co-crystal of a compound of formula 2, wherein

R^2aIs H, C_1-4Alkyl radical, C_1-4-alkynyl, C_3-6-cycloalkyl, O-C_1-4Alkyl, CONR^2a.1R^2a.2；

R^2a.1Is C_1-4-an alkyl group;

R^2a.2is H;

R^2bis H, C_1-4Alkyl, O-C_1-4Alkyl, CONR^2b.1R^2b.2；

R^2b.1Is C_1-4Alkyl radical, C_0-4-alkyl-C_3-6-cycloalkyl, C_1-4-a haloalkyl group;

R^2b.2is H, C_1-4-an alkyl group;

or R^2b.1And R^2b.2Together are C_3-6-alkylene groups forming a heterocyclic ring with the nitrogen atom, wherein optionally one carbon atom of the ring is replaced by an oxygen atom and the remaining residues are as defined above.

Another aspect of the invention also includes administering to a subject a co-crystal of a compound of formula 2, wherein

R^2aIs H, C_1-4-an alkyl group,

R^2bis H, CONR^2b.1R^2b.2；

R^2b.1Is C_1-4Alkyl radical, C_0-4-alkyl-C_3-6-cycloalkyl, C_1-4-a haloalkyl group;

R^2b.2is H, C_1-4-an alkyl group;

or R^2b.1And R^2b.2Together are C_3-6Alkylene radical ofForm a heterocyclic ring with the nitrogen atom, wherein optionally one carbon atom of the ring is replaced by an oxygen atom and the remaining residues are as defined above.

Another aspect of the invention also includes administering to a subject a co-crystal of a compound of formula 2, wherein

R¹Is C_1-6Alkyl radical, C_1-6Haloalkyl, O-C_1-6-haloalkyl, halogen;

m is 1 or 2;

R^2ais H, C_1-4-an alkyl group;

R^2bis H, CONR^2b.1R^2b.2；

R^2b.1Is C_1-4Alkyl radical, C_0-4-alkyl-C_3-6-cycloalkyl, C_1-4-a haloalkyl group;

R^2b.2is H, C_1-4-an alkyl group;

or R^2b.1And R^2b.2Together are C_3-6-alkylene, which forms a heterocyclic ring with the nitrogen atom, wherein optionally one carbon atom of the ring is replaced by an oxygen atom;

R³Is H, C_1-6-an alkyl group;

x is an anion selected from chloride or dibenzoyl tartrate;

j is 1 or 2.

Another aspect of the invention also includes administering to a subject a co-crystal of a compound of formula 2, wherein

R^2aIs H, C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2bis H, CONR^2b.1R^2b.2；

R^2b.1Is C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2b.2is C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

and the remaining residues are as defined above.

Another aspect of the invention also includes administering to a subject a co-crystal of a compound of formula 2, wherein

R^2aIs H, C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2bis H, CONR^2b.1R^2b.2；

R^2b.1Is C_0-4-alkyl-C_3-6-a cycloalkyl group;

R^2b.2is H, C_1-4-an alkyl group; h, methyl, ethyl, propyl in some examples;

and the remaining residues are as defined above.

Another aspect of the invention also includes administering to a subject a co-crystal of a compound of formula 2, wherein

R^2aIs H, C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2bis H, CONR^2b.1R^2b.2；

R^2b.1Is C_1-4-a haloalkyl group;

R^2b.2is H, C_1-4-an alkyl group; h, methyl, ethyl, propyl in some examples;

and the remaining residues are as defined above.

Another aspect of the invention also includes administering to a subject a co-crystal of a compound of formula 2, wherein

R^2b.1And R^2b.2Together are C_3-6-alkylene groups forming a heterocyclic ring with the nitrogen atom, wherein optionally one carbon atom of the ring is replaced by an oxygen atom and the remaining residues are as defined above.

Another aspect of the invention also includes administering to the subject a co-crystal of a compound of formula 2, wherein R is¹、m、R^2a、R^2b、R³X and j are as defined above and the co-crystal former is selected from: ascorbic acid, mucic acid, pamoic acid, succinamide, nicotinic acid, nicotinamide, isonicotinamide, l-lysine, l-proline or their hydrates or hydrochlorides.

Another aspect of the invention also includes administering to the subject a co-crystal of a compound of formula 2a, wherein R^2a、R^2b、R³X and j are as defined above

Yet another aspect of the invention includes administering to a subject a co-crystal of a compound of formula 2a, wherein

R^2aIs H, C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2bis H, CONR^2b.1R^2b.2；

R^2b.1Is C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2b.2is C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

and the remaining residues are as defined above.

Yet another aspect of the invention includes administering to a subject a co-crystal of a compound of formula 2a, wherein

R^2aIs H, C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2bis H, CONR ^2b.1R^2b.2；

R^2b.1Is C_0-4-alkyl-C_3-6-a cycloalkyl group;

R^2b.2is H, C_1-4-an alkyl group; h, methyl, ethyl, propyl in some examples;

and the remaining residues are as defined above.

Yet another aspect of the invention includes administering to a subject a co-crystal of a compound of formula 2a, wherein

R^2aIs H, C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2bis H, CONR^2b.1R^2b.2；

R^2b.1Is C_1-4-a haloalkyl group;

R^2b.2is H, C_1-4-an alkyl group; h, methyl, ethyl, propyl in some examples;

and the remaining residues are as defined above.

Yet another aspect of the invention includes administering to a subject a co-crystal of a compound of formula 2a, wherein

R^2b.1And R^2b.2Together are C_3-6-alkylene, which forms a heterocyclic ring with the nitrogen atom, wherein optionally one carbon atom of the ring is replaced by an oxygen atom;

and the remaining residues are as defined above.

The free base of the compound of formula 2 (j ═ 0) is typically amorphous and is used in the process of making the co-crystal, although in some examples the process of making the co-crystal employs a salt of the compound of formula 2. Thus, another aspect of the invention is a salt of a compound of formula 2, wherein R is¹、m、R^2a、R^2b、R³Are as defined above for these co-crystals and

x is an anion selected from chloride, bromide, iodide, sulfate, phosphate, methanesulfonate, nitrate, maleate, acetate, benzoate, citrate, salicylate, fumarate, tartrate, dibenzoyltartrate, oxalate, succinate, benzoate, and p-toluenesulfonate; in some examples, chloride or dibenzoyltartrate;

j is 0, 0.5, 1, 1.5 or 2; in some instances 1 or 2.

Another aspect of the invention also includes administering to the subject a co-crystal of a compound of formula 2, wherein R is¹、m、R^2a、R^2b、R³Are as defined above for these co-crystals and

x is an anion selected from chloride or dibenzoyl tartrate;

j is 1 or 2.

Another aspect of the present invention also includes administering to a subject a salt of a compound of formula 2, wherein R¹、m、R^2a、R^2b、R³Are as defined above for these salts, and X is chloride and j is 2.

Another aspect of the present invention also includes administering to a subject a salt of a compound of formula 2, wherein R¹、m、R^2a、R^2b、R³Are as defined above for these salts, and X is dibenzoyltartrate and j is 1.

Another aspect of the invention also includes administering to the subject a salt of a compound of formula 2a, wherein R^2a、R^2b、R³X and j are as defined above

Yet another aspect of the present invention includes administering to a subject a salt of a compound of formula 2a, wherein

R^2aIs H, C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2bis H, CONR^2b.1R^2b.2；

R^2b.1Is C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2b.2is C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

and the remaining residues are as defined above.

Yet another aspect of the present invention includes administering to a subject a salt of a compound of formula 2a, wherein

R^2aIs H, C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2bis H, CONR^2b.1R^2b.2；

R^2b.1Is C_0-4-alkyl-C_3-6-a cycloalkyl group;

R^2b.2is H, C_1-4-an alkyl group; h, methyl, ethyl, propyl in some examples;

and the remaining residues are as defined above.

Yet another aspect of the present invention includes administering to a subject a salt of a compound of formula 2a, wherein

R^2aIs H, C_1-4-an alkyl group; methyl, ethyl, propyl in some examples;

R^2bis H, CONR^2b.1R^2b.2；

R^2b.1Is C_1-4-a haloalkyl group;

R^2b.2is H, C_1-4-an alkyl group; h, methyl, ethyl, propyl in some examples;

and the remaining residues are as defined above.

Yet another aspect of the present invention includes administering to a subject a salt of a compound of formula 2a, wherein

R^2b.1And R^2b.2Together are C_3-6-alkylene groups forming a heterocyclic ring with the nitrogen atom, wherein optionally one carbon atom of the ring is replaced by an oxygen atom and the remaining residues are as defined above.

Another aspect of the invention also includes administering to the subject a salt of a compound of formula 2a, wherein R¹、m、R^2a、R^2b、R³Are as defined above for these salts, and X is chloro and j is 2.

Another aspect of the invention also includes administering to the subject a salt of a compound of formula 2a, wherein R¹、m、R^2a、R^2b、R³Are as defined above for these salts, and X is dibenzoyltartrate and j is 1. Another aspect of the invention is a salt of a compound of formula 2a, wherein R ¹、m、R^2a、R^2b、R³Are as defined above for these salts, and X is (S) - (S) - (+) -2, 3-dibenzoyl-tartrate and j is 1.

c. Preparation

Other embodiments of the invention also include administering to a subject a pharmaceutical composition comprising a compound of formula 3

Wherein

R¹Is H, C_1-6Alkyl radical, C_0-4-alkyl-C_3-6-cycloalkyl, C_1-6-a haloalkyl group;

R²is H, C_1-6-an alkyl group;

x is an anion selected from chloride or 1/2 dibenzoyl tartrate;

j is 1 or 2.

An embodiment of the present invention further comprises administering to a subject a pharmaceutical composition comprising a compound of formula 3, wherein

R¹Is H, C_1-6-an alkyl group;

R²is H, C_1-6-an alkyl group;

x is an anion selected from chloride or 1/2 dibenzoyl tartrate;

j is 1 or 2.

An embodiment of the present invention further comprises administering to a subject a pharmaceutical composition comprising a compound of formula 3, wherein

R¹H, methyl, ethyl, propyl, butyl;

R²h, methyl, ethyl, propyl, butyl;

x is an anion selected from chloride or 1/2 dibenzoyl tartrate, such as chloride;

j is 1 or 2, and in some examples is 2.

An embodiment of the present invention further comprises administering to a subject a pharmaceutical composition comprising a compound of formula 3, wherein

R¹H, methyl, ethyl, propyl, butyl;

R²H and methyl;

x is an anion selected from chloride or 1/2 dibenzoyl tartrate, such as chloride;

j is 1 or 2, and in some examples is 2.

An embodiment of the present invention further comprises administering to a subject a pharmaceutical composition comprising a compound of formula 3, wherein

R¹H and methyl;

R²h and methyl;

x is an anion selected from chloride or 1/2 dibenzoyl tartrate, such as chloride;

j is 1 or 2, and in some examples is 2.

An embodiment of the invention also includes administering to the subject a pharmaceutical composition containing a compound described in table 2 as a hydrochloride salt. Other embodiments of the invention also include administering to the subject a pharmaceutical composition comprising a compound described in table 2 as the dihydrochloride salt.

TABLE 2

Another object of the invention is a pharmaceutical dosage form for administering the above compound to a subject, wherein the dosage form is an oral delivery dosage form.

Another object of the invention is a pharmaceutical dosage form for administering the above compound to a subject, in the form of a tablet, capsule, pill, powder or granule.

It is another object of the present invention to administer to a subject a pharmaceutical dosage form as described above for use as a medicament.

Another object of the present invention is the use of a pharmaceutical dosage form as described above for the preparation of a medicament for the treatment of a neurodegenerative disease or disorder selected from alzheimer's disease, parkinson's disease, frontotemporal dementia, huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, vascular dementia, and progressive supranuclear palsy.

Another object of the present invention is a method for the treatment and/or prevention of a disease or disorder selected from neurodegenerative diseases such as alzheimer's disease, parkinson's disease, frontotemporal dementia, huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, vascular dementia, and progressive supranuclear palsy, characterized by orally administering to a subject or patient an effective amount of a pharmaceutical dosage form as defined above once, twice, three times or several times daily.

d. Dosage forms/compositions

Solid pharmaceutical compositions ready to use/ingest made from the compound of formula 3 include powders, granules, pills, lozenges, capsules, chewable lozenges, dispersible lozenges, tablets and buccal agents. In detail:

the capsule formulation of the invention comprises the powdered intermediate of the compound of formula 3 (intermediate blend comprising the powdered intermediate), pellets or granules obtained by known wet, dry or hot melt granulation or hot melt extrusion or spray drying of suitable intermediate blends, filled in known capsules (e.g. hard gelatin or HPMC capsules).

The above capsule formulation may also comprise a powdered intermediate of the compound of formula 3 in compacted form.

The capsule formulation of the invention comprises a compound of formula 3 suspended or diluted in a liquid or liquid mixture.

The lozenge formulations of the invention comprise such lozenges obtained by compression of a suitable final blend or by pastillation of pellets or granules obtained by known wet, dry or hot melt granulation or hot melt extrusion or spray drying of a suitable intermediate blend.

Another object of the present invention is a dosage form wherein a pH adjuster or buffer is added for improved stability of the active ingredient. The pH adjustor/buffer may be a basic amino acid having amino and base properties (isoelectric point, pI: 7.59 to 10.76), such as, for example, L-arginine, L-lysine or L-histidine. The buffer in the sense of the present invention is L-arginine. L-arginine has a particularly suitable stabilizing effect on the compositions of the invention, for example, by inhibiting chemical degradation of the compound of formula 3.

Thus, in one embodiment, the present invention relates to a pharmaceutical composition (e.g., an oral solid dosage form, particularly a lozenge) comprising a compound of formula 3 and L-arginine for stabilizing the composition, particularly against chemical degradation; and one or more than one pharmaceutical excipient.

Suitably, the pharmaceutical excipients used in the present invention are known substances such as cellulose and its derivatives, D-mannitol, corn starch, pregelatinized starch as filler, copovidone as binder, crospovidone as disintegrant, magnesium stearate as lubricant, colloidal anhydrous silicon dioxide as glidant, hypromellose as film coating agent, polyethylene glycol as plasticizer, titanium dioxide, red/yellow iron oxide as pigment, talc and the like.

In detail, the pharmaceutical excipient may be first and second diluents, binders, disintegrants and lubricants; additional disintegrants and additional glidants are another option.

Suitable diluents for the pharmaceutical compositions according to the invention are cellulose powder, microcrystalline cellulose, lactose modified in various crystalline forms, anhydrous dibasic calcium phosphate, dibasic calcium phosphate dihydrate, erythritol, low-substituted hydroxypropylcellulose, mannitol, starch or modified starch (e.g. pregelatinized or partially hydrolyzed), or xylitol. Mannitol and microcrystalline cellulose in those diluents are employed in some examples.

The diluents used as the second diluent are mannitol and microcrystalline cellulose, which are the diluents mentioned above.

Lubricants suitable for the pharmaceutical compositions according to the invention are talc, polyethylene glycol, calcium behenate, calcium stearate, sodium stearyl fumarate, hydrogenated castor oil or magnesium stearate. In some examples the lubricant is magnesium stearate.

Binders suitable for the pharmaceutical compositions of the invention are copovidone (copolymer of vinylpyrrolidone with other vinyl derivatives), Hydroxypropylmethylcellulose (HPMC), Hydroxypropylcellulose (HPC), polyvinylpyrrolidone (povidone), pregelatinized starch, stearic-palmitic acid, low-substituted hydroxypropylcellulose (L-HPC), copovidone and pregelatinized starch being used in some formulations. The binders pregelatinized starch and L-HPC described above exhibit additional diluent and disintegrant properties, and may also be used as a second diluent or disintegrant.

Suitable disintegrants for the pharmaceutical compositions according to the invention are corn starch, crospovidone, polacrilin potassium (polacrilin potassium), croscarmellose sodium, low-substituted hydroxypropylcellulose (L-HPC) or pregelatinized starch; such as croscarmellose sodium.

Colloidal silica may be used as an optional slip aid.

Exemplary compositions of the invention comprise mannitol as a diluent, microcrystalline cellulose as a diluent with additional disintegration properties, binder copovidone, croscarmellose sodium as a disintegrant, and magnesium stearate as a lubricant.

A typical pharmaceutical composition comprises (% by weight)

The pharmaceutical composition according to some embodiments comprises (% by weight)

The pharmaceutical composition according to some embodiments comprises (% by weight)

The pharmaceutical composition according to some embodiments comprises (% by weight)

The pharmaceutical composition according to some embodiments comprises (% by weight)

In some examples pharmaceutical compositions containing 10 to 90% active ingredient, such as 30 to 70% active ingredient (% by weight) are used.

Lozenge formulations of the present invention are uncoated or coated (e.g., film coated) with a suitable coating known to not adversely affect the dissolution properties of the final formulation. For example, tablets with a sealed coating may be provided by dissolving a high molecular weight polymer such as polyvinylpyrrolidone or hydroxypropylmethylcellulose together with a plasticizer, a lubricant and optionally pigments and surfactants in water or an organic solvent such as acetone and spraying this mixture onto the tablet core in a coating apparatus such as a pan coater or a fluidized bed coater equipped with a wurster insert, for the protection of the patient environment and clinical workmen and for moisture protection purposes.

In addition, agents such as beeswax, shellac, cellulose acetate phthalate, polyvinyl acetate phthalate, zein (zein), film forming polymers such as hydroxypropyl cellulose, ethyl cellulose and polymeric methacrylates can be applied to the lozenge, provided that the coating does not materially affect the disintegration/dissolution of the dosage form and the stability of the coated dosage form is not affected.

After the dosage form is film coated, a sugar coating may be applied to the sealed pharmaceutical dosage form. The glaze may comprise sucrose, dextrose, sorbitol, and the like, or mixtures thereof. If desired, colorants or opacifiers may be added to the sugar solution.

The solid preparation of the present invention has a tendency to absorb moisture. They can be packaged using PVC blisters, PVDC blisters or moisture-proof packaging materials such as aluminum foil blister packs, Alu/Alu blisters, transparent or opaque polymer blisters with bags, polypropylene tubes, glass bottles and HDPE bottles, optionally with child-resistance or tamper-resistance. The primary packaging material may contain a desiccant, such as a molecular sieve or silica gel, to improve the chemical stability of the API. Opaque packaging (such as colored blister materials, tubes, brown glass bottles, or the like) can extend the shelf life of an API by reducing photodegradation.

e. Dosage form

The dosage range of the compound of formula 3 is usually 100 to 1000mg, in particular 200 to 900mg, 300 to 900mg or 350 to 850mg or 390 to 810 mg. One or two lozenges may be provided, wherein in some examples two lozenges are used for a daily oral dose of 100, 200, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900mg, and in some examples a daily oral dose of 350, 400, 450, 750, 800, 850 mg.

Dosage ranges can be achieved by one tablet or by two tablets; in some examples, two lozenges are administered, each lozenge containing half of the dose.

The active ingredient may be administered up to three times a day, such as once or twice a day. The specific dose concentration is 400mg or 800 mg.

f. Terms and definitions used

Terms not explicitly defined herein shall have their meanings as given by the disclosure and content to these terms by those skilled in the art. However, as used in this specification, unless specified to the contrary, the following terms have the meanings indicated and are accompanied by the following conventions.

The term "about" means more or less than 5% of the stated value. Thus, about 100 minutes can also be read as 95 to 105 minutes.

In the case where the compound of the present invention is expressed in the form of a chemical name and in the form of a chemical formula, the chemical formula should be generalized in order to prevent any deviation. The asterisks used in the substoichiometric formulae indicate the bond to which the defined parent molecule is attached.

Unless specifically indicated, throughout the specification and the appended claims, a given chemical formula or name shall encompass tautomers and all stereoisomers, optical and geometric isomers (e.g., enantiomers, diastereomers, E/Z isomers, etc.) and racemates thereof as well as mixtures of individual enantiomers in varying proportions, mixtures of diastereomers, or mixtures of any of the foregoing forms in which these isomers and enantiomers are present, as well as salts, including pharmaceutically acceptable salts and solvates thereof (such as, for example, hydrates, solvates including free compounds or solvates of salts of such compounds).

As used herein, the term "substituted" means that any one or more hydrogens on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valence is not exceeded, and that the substitution results in a stable compound.

Within the scope of the present invention, the term "optionally substituted" means that the aforementioned groups are optionally substituted by low molecular weight groups. Examples of low molecular groups considered to have chemical significance are groups consisting of 1 to 200 atoms. Of interest are such groups that do not negatively impact the pharmacological efficacy of the compound. For example, these groups may comprise:

Linear or branched carbon chains, optionally interrupted by heteroatoms, optionally substituted by rings, heteroatoms or other common functional groups.

Aromatic or non-aromatic ring systems consisting of carbon atoms and optionally heteroatoms, which may in turn be substituted by functional groups.

A plurality of aromatic or non-aromatic ring systems consisting of carbon atoms and optionally heteroatoms, which may be linked via one or more carbon chains, optionally interrupted by heteroatoms, optionally substituted by heteroatoms or other customary functional groups.

The compounds disclosed herein may be present as pharmaceutically acceptable salts. The invention includes the compounds listed above in salt (including acid addition salts) form. Suitable salts include those formed with organic and inorganic acids. These acid addition salts will generally be pharmaceutically acceptable. However, salts of non-pharmaceutically acceptable salts may be used in the preparation and purification of the compounds. Base addition salts may also be formed and are pharmaceutically acceptable. For a more complete discussion of salt preparation and Selection, see Pharmaceutical Salts, Properties, Selection, and Use (Stahl, P.Heinrich.Wiley-VCHA, Zurich, Switzerland, 2002).

The term "pharmaceutically acceptable salt" as used herein means a salt or zwitterionic form of a compound disclosed herein, which is water-or oil-soluble or dispersible and therapeutically acceptable as defined herein. Salts may be prepared during the final isolation and purification of the compounds or separately by reacting the appropriate compound in its free base form with an appropriate acid. Representative acid addition salts include acetate, adipate, alginate, L-ascorbate, aspartate, benzoate, benzenesulfonate (benzanesulfonate/besylate), bisulfate, butyrate, camphorate, camphorsulfonate, citrate, bisgluconate, formate, fumarate, gentisate, glutarate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate (isethionate), lactate, maleate, malonate, DL-mandelate, trimethylbenzenesulfonate, methanesulfonate, naphthalenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectate, persulfate, 3-phenylpropionate, phosphonate, picrate, etc, Pivalate, propionate, pyroglutamate, succinate, sulfonate, tartrate, L-tartrate, trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate, p-toluenesulfonate (tolumenesulfonate/p-tosylate), and undecanoate. Likewise, the basic groups in the compounds disclosed herein can be substituted with methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl, dibutyl and diamyl sulfates; decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; and quaternary ammonification of benzyl and phenethyl bromides. Examples of acids that may be used to form therapeutically acceptable addition salts include inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid; and organic acids such as oxalic acid, maleic acid, succinic acid, and citric acid. It can also be formed by coordinating a compound with an alkali metal or alkaline earth metal ion. Accordingly, the present invention encompasses sodium, potassium, magnesium, and calcium salts, and the like, of the compounds disclosed herein.

Base addition salts can be prepared during the final isolation and purification of the compounds by reacting the carboxyl group with a suitable base, such as a hydroxide, carbonate or bicarbonate of a metal cation, or with ammonia or an organic first, second or third amine. Cations of therapeutically acceptable salts include lithium, sodium, potassium, calcium, magnesium, and aluminum, as well as nontoxic quaternary ammonium cations such as ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, N-dibenzylphenethylamine, 1-amphetamine, and N, P-dibenzylethylenediamine. Other representative organic amines suitable for use in forming base addition salts include ethylenediamine, ethanolamine, diethanolamine, piperidine, and piperazine.

Although the compounds of the present invention may be administered as a chemical source, they may also be provided in the form of pharmaceutical preparations. Accordingly, provided herein are pharmaceutical formulations comprising one or more of certain compounds disclosed herein, or one or more pharmaceutically acceptable salts, esters, prodrugs, amides, or solvates thereof, and one or more pharmaceutically acceptable carriers thereof and optionally one or more other therapeutic ingredients. The carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof. Suitable formulations depend on the chosen route of administration. May be suitably and as such; for example, Remington's Pharmaceutical Sciences understand the use of any of the well-known techniques, carriers, and excipients. The pharmaceutical compositions disclosed herein can be manufactured in a manner known in the art, for example, by means of known mixing, dissolving, granulating, dragee-making, wet-milling (levigating), emulsifying, encapsulating, entrapping or compressing processes.

"heterocycle" ("het") includes a five-, six-, or seven-membered saturated or unsaturated heterocycle or a 5-to 10-membered bicyclic heterocycle which may contain one, two, or three heteroatoms selected from oxygen, sulfur, and nitrogen; the ring may be attached to the molecule via a carbon atom or, if present, via a nitrogen atom. The following are examples of five-, six-or seven-membered saturated or unsaturated heterocycles:

unless otherwise specified, a heterocycle having a keto group may be provided. Examples include:

examples of 5-to 10-membered bicyclic heterocycles are pyrrolizine, indole, indolizine, isoindole, indazole, purine, quinoline, isoquinoline, benzimidazole, benzofuran, benzopyran, benzothiazole, benzisothiazole, pyridopyrimidine, pteridine, pyrimidopyrimidine, dihydrobenzoxazole,

while the term heterocycle includes heterocyclic aromatic groups, the term heterocyclic aromatic group ("heteroaryl") denotes a five or six membered heterocyclic aromatic group or a 5 to 10 membered bicyclic heteroaryl ring which may contain one, two or three heteroatoms selected from oxygen, sulfur and nitrogen, which contains sufficient conjugated double bonds to form an aromatic system. The ring may be attached to the molecule through a carbon atom or, if present, through a nitrogen atom. The following are examples of five-or six-membered heterocyclic aromatic groups:

Examples of 5-to 10-membered bicyclic heteroaryls are pyrrolizine, indole, indolizine, isoindole, indazole, purine, quinoline, isoquinoline, benzimidazole, benzofuran, benzopyran, benzothiazole, benzisothiazole, pyridopyrimidine, pteridine, pyrimidopyrimidine.

As used herein, the term "halogen" means a halogen substituent selected from fluorine, chlorine, bromine or iodine.

The term "C_1-6Alkyl (including those which are part of other groups) means branched and unbranched alkyl having from 1 to 6 carbon atoms, and the term "C_1-4By alkyl "is meant branched and unbranched alkyl groups having from 1 to 4 carbon atoms. In some instancesIn an alkyl group having 1 to 4 carbon atoms. Examples of such alkyl groups include: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, second butyl, third butyl, n-pentyl, isopentyl, neopentyl or hexyl. The groups may also optionally be represented by the abbreviations Me, Et, n-Pr, i-Pr, n-Bu, i-Bu, t-Bu, etc. Unless otherwise indicated, the definitions propyl, butyl, pentyl and hexyl include all possible isomeric forms of the groups. Thus, for example, propyl includes n-propyl and isopropyl, and butyl includes isobutyl, second butyl, third butyl, and the like.

The term "C_1-6Alkylene "(including those which are part of other groups) means branched and unbranched alkylene groups having from 1 to 6 carbon atoms, and the term" C_1-4By alkylene is meant branched and unbranched alkylene groups having 1 to 4 carbon atoms. In some examples, alkylene groups having 1 to 4 carbon atoms are present. Examples include: methylene, ethylene, propylene, 1-methylethylene, butylene, 1-methylpropylene, 1-dimethylethylene, 1, 2-dimethylethylene, pentylene, 1-dimethylpropylene, 2-dimethylpropylene, 1, 3-dimethylpropylene or hexylene. Unless otherwise indicated, the definitions propylene, butylene, pentylene and hexylene also include all possible isomeric forms of the relevant groups having the same carbon number. Thus, for example, propyl also includes 1-methylethylene and butylene includes 1-methylpropylene, 1-dimethylethylene, 1, 2-dimethylethylene.

The term "C_2-6Alkenyl "(including those which are part of other groups) denotes branched and unbranched alkenyl groups having 2 to 6 carbon atoms, and the term" C_2-4-alkenyl "denotes branched and unbranched alkenyl groups having 2 to 4 carbon atoms, with the proviso that they have at least one double bond. Alkenyl groups having 2 to 4 carbon atoms are used in some examples. Examples include: vinyl (ethenyl or vinyl), propenyl, butenyl, pentenyl or hexenyl. Unless otherwise indicated, the definitions propenyl, butenyl, pentenyl and hexenyl include all possible isomeric forms of the groups. Thus, for example, C The alkenyl group includes 1-propenyl and 2-propenyl, and the butenyl group includes 1-, 2-and 3-butenyl, 1-methyl-1-propenyl, 1-methyl-2-propenyl, and the like.

The term "C_2-6-alkenylene "(including those that are part of other groups) means branched and unbranched alkenylene having 2 to 6 carbon atoms, and the term" C_2-4By alkenylene "is meant branched and unbranched alkenylene groups having 2 to 4 carbon atoms. In some examples, alkenylene groups having 2 to 4 carbon atoms are present. Examples include: vinylidene, propenylene, 1-methylvinylene, butenylene, 1-methylpropylene, 1-dimethylvinylene, 1, 2-dimethylvinylene, pentenylene, 1-dimethylpropenyl, 2-dimethylpropenyl, 1, 3-dimethylpropenyl or hexenylene. Unless otherwise indicated, the definitions propenylene, butenylene, pentenylene and hexenylene include all possible isomeric forms of each group having the same carbon number. Thus, for example, propenyl also includes 1-methylethenylene, and butenylene includes 1-methylpropenylene, 1-dimethylethenylene, 1, 2-dimethylethenylene.

The term "C_2-6Alkynyl (including those which are part of other groups) means branched and unbranched alkynyl having 2 to 6 carbon atoms, and the term "C_2-4By alkynyl is meant branched and unbranched alkynyl groups having 2 to 4 carbon atoms, with the proviso that they have at least one triple bond. In some examples, alkynyl groups having 2 to 4 carbon atoms are present. Examples include: ethynyl, propynyl, butynyl, pentynyl or hexynyl. Unless otherwise indicated, the definitions propynyl, butynyl, pentynyl and hexynyl include all possible isomeric forms of the respective radicals. Thus, for example, propynyl includes 1-propynyl and 2-propynyl, butynyl includes 1-, 2-, and 3-butynyl, 1-methyl-1-propynyl, 1-methyl-2-propynyl, and the like.

The term "C_2-6-alkynylene "(including those that are part of other groups) means branched and unbranched alkynylene groups having 2 to 6 carbon atoms, and the term" C_2-4-alkynylene "means having 2 toBranched and unbranched alkynylene of 4 carbon atoms. In some examples, alkynylene groups having 2 to 4 carbon atoms are present. Examples include: ethynylene, propynyl, 1-methylacetylene, butynyl, 1-methylpropynyl, 1-dimethylethynylene, 1, 2-dimethylethynylene, pentynyl, 1-dimethylpropynyl, 2-dimethylpropynyl, 1, 3-dimethylpropynyl or hexynyl. Unless otherwise indicated, the definitions propynyl, butynyl, pentynyl and hexynyl include all possible isomeric forms of the respective radicals having the same carbon number. Thus, for example, propynyl also includes 1-methylacetylene, and butynyl includes 1-methylpropynyl, 1-dimethylethyleneene, 1, 2-dimethylethyleneene.

As used herein, the term "C_3-6-cycloalkyl "(including those that are part of other groups) means cycloalkyl groups having 3 to 6 carbon atoms, where in some examples these groups are cycloalkyl groups having 5 to 6 carbon atoms. Examples include: cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

The term "C_1-6Haloalkyl "(including those which are part of other groups) means branched and unbranched alkyl groups having 1 to 6 carbon atoms in which one or more than one hydrogen atom is replaced by a halogen atom selected from fluorine, chlorine or bromine, such as fluorine and chlorine, e.g. fluorine. The term "C_1-4Haloalkyl "means branched and unbranched alkyl groups corresponding to 1 to 4 carbon atoms in which one or more than one hydrogen atom is replaced as described above. In some instances, C is present_1-4-haloalkyl. Examples include: CH (CH)₂F、CHF₂、CF₃。

The term "C" alone or in combination with another group_1-n-alkyl "(where n is an integer from 2 to n) represents a non-cyclic saturated branched or linear hydrocarbon radical having from 1 to n C atoms. For example the term C_1-5Alkyl includes the group H₃C-、H₃C-CH₂-、H₃C-CH₂-CH₂-、H₃C-CH(CH₃)-、H₃C-CH₂-CH₂-CH₂-、H₃C-CH₂-CH(CH₃)-、H₃C-CH(CH₃)-CH₂-、H₃C-C(CH₃)₂-、H₃C-CH₂-CH₂-CH₂-CH₂-、H₃C-CH₂-CH₂-CH(CH₃)-、H₃C-CH₂-CH(CH₃)-CH₂-、H₃C-CH(CH₃)-CH₂-CH₂-、H₃C-CH₂-C(CH₃)₂-、H₃C-C(CH₃)₂-CH₂-、H₃C-CH(CH₃)-CH(CH₃) -and H₃C-CH₂-CH(CH₂CH₃)-。

The term "C" alone or in combination with another group_1-nHaloalkyl "(wherein n is an integer from 2 to n) denotes an acyclic saturated branched or linear hydrocarbon radical having 1 to n C atoms, wherein one or more hydrogen atoms are replaced by halogen atoms selected from fluorine, chlorine or bromine, such as fluorine and chlorine, e.g. fluorine. Examples include: CH (CH) ₂F、CHF₂、CF₃。

The term "C" alone or in combination with another group_1-nAlkylene "(where n is an integer from 2 to n) represents a non-cyclic linear or branched divalent alkyl radical containing from 1 to n C atoms. For example the term C_1-4Alkylene includes-CH₂-、-CH₂-CH₂-、-CH(CH₃)-、-CH₂-CH₂-CH₂-、-C(CH₃)₂-、-CH(CH₂CH₃)-、-CH(CH₃)-CH₂-、-CH₂-CH(CH₃)-、-CH₂-CH₂-CH₂-CH₂-、-CH₂-CH₂-CH(CH₃)-、-CH(CH₃)-CH₂-CH₂-、-CH₂-CH(CH₃)-CH₂-、-CH₂-C(CH₃)₂-、-C(CH₃)₂-CH₂-、-CH(CH₃)-CH(CH₃)-、-CH₂-CH(CH₂CH₃)-、-CH(CH₂CH₃)-CH₂-、-CH(CH₂CH₂CH₃)-、-CH(CH(CH₃))₂-and-C (CH)₃)(CH₂CH₃)-。

The term "C_2-n-alkenyl "for compounds having at least two carbon atoms as defined for" C_1-n-alkyl "as long as at least two of those carbon atoms of the group are bonded to each other via a double bond.

The term "C_2-nAlkynyl for a radical having at least two carbon atoms as defined for "C_1-n-alkyl "as long as at least two of those carbon atoms of the group are bonded to each other via a triple bond.

The term "C" alone or in combination with another group_3-n-cycloalkyl "(wherein n is an integer from 4 to n) represents a cyclic saturated unbranched hydrocarbon radical having 3 to n C atoms. For example, the term C_3-7Cycloalkyl includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

It should be noted that, as used herein and in the appended claims, no element preceding an element contains a plural reference unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells, and reference to "a peptide" includes reference to one or more peptides and equivalents thereof (e.g., polypeptides) known to those skilled in the art, and so forth.

By "an individual suffering from or at risk of suffering from age-related cognitive impairment" is meant an individual who has spent more than about 50% of their life expectancy, such as more than 60%, for example, more than 70%, such as more than 75%, 80%, 85%, 90%, 95% or even 99%. The age of the individual will depend on the species. Thus, this percentage is based on the predicted life expectancy of the species. For example, in humans, such individuals are 50 years or older, e.g., 60 years or older, 70 years or older, 80 years or older, 90 years or older, and typically no greater than 100 years old, such as 90 years old, i.e., at any age from about 50 to 100 years old, e.g., 50 … … 55 … … 60 … … 65 … … 70 … … 75 … … 80 … … 85 … … 90 … … 95 … … 100 years old or 50 to 1000, who are suffering from a senescence-associated disorder as further described below, e.g., cognitive impairment associated with the natural senescence process; an individual 50 years of age or older, e.g., 60 years of age or older, 70 years of age or older, 80 years of age or older, 90 years of age or older, and typically no older than 100 years of age, i.e., who has not begun to display symptoms of a condition associated with aging (e.g., cognitive impairment) at about 50 and 100 years of age, e.g., 50 … … 55 … … 60 … … 65 … … 70 … … 75 … … 80 … … 85 … … 90 … … 95 … … 100 years of age 100; individuals of any age suffering from cognitive impairment due to age-related disorders as described further below, and individuals of any age diagnosed with age-related disorders that are typically accompanied by cognitive impairment, wherein the individual has not yet begun to exhibit symptoms of cognitive impairment. The corresponding age of the non-human subject is known and intended for use herein.

As summarized elsewhere, in some examples, the subject is a mammal. Mammalian species that can be treated by the present method include dogs and cats; a horse; cattle; sheep; etc., and primates, including humans. For example, in experimental studies, the subject methods, compositions, and reagents may also be applied to animal models, including small mammals, such as rodents, lagomorphs, and the like.

As used herein and as described above, "treating" refers to any of (i) preventing a disease or disorder, or (ii) reducing or eliminating a symptom of a disease or disorder. Treatment can be performed either prophylactically (prior to the onset of the disease) or therapeutically (after the onset of the disease). The effect may be prophylactic in terms of completely or partially preventing a disease or a symptom thereof and/or may be therapeutic in terms of a partial or complete cure of a disease and/or adverse effects due to the disease. Thus, as used herein, the term "treatment" encompasses any treatment of an aging-related disease or disorder in a mammal, and includes: (a) preventing the occurrence of a disease in an individual, who may be predisposed to the disease but has not yet been diagnosed with the disease; (b) inhibiting the disease, i.e., inhibiting its development; or (c) relieving the disease, i.e., causing disease regression. Treatment may result in a variety of different physical manifestations, e.g., modulation of gene expression, tissue or organ rejuvenation, etc. The therapeutic agent may be administered before, during or after onset of the disease. Of particular interest is the treatment of ongoing diseases, wherein the treatment stabilizes or reduces the patient's undesirable clinical symptoms. These treatments may be performed before the affected tissue is completely dysfunctional. The targeted therapy may be administered at, and in some cases after, the symptomatic phase of the disease.

In some embodiments, the disorder treated is impairment of cognitive ability associated with aging in the individual. Cognitive ability or "cognition" means mental processes that include attention and concentration, learning complex tasks and concepts, memory (short and/or long term acquisition, retention and retrieval of new information), information processing (processing of information collected by five senses), visual spatial functions (vision, depth perception, use of mental imagery, copying, object or shape construction), language production and understanding, verbal fluency (vocabulary discovery), problem solving, decision making, and executive functions (planning and prioritization). By "cognitive decline" is meant a progressive decline in one or more of these abilities, e.g., decline in memory, language, thinking, judgment, etc. By "cognitive impairment" and "cognitive impairment" is meant a decrease in cognitive ability relative to a healthy individual (e.g., an age-matched healthy individual) or relative to the individual's ability at an earlier time point (e.g., 2 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 2 years, 5 years, or 10 years or earlier). By "aging-related cognitive impairment" is meant impairment of cognitive ability normally associated with aging, including, for example, cognitive impairment associated with the natural aging process, e.g., mild cognitive impairment (m.c.i.); and cognitive impairment associated with aging-related disorders, i.e., disorders that increase in frequency with increasing aging, e.g., neurodegenerative disorders such as alzheimer's disease, parkinson's disease, frontotemporal dementia, huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, vascular dementia, progressive supranuclear palsy, ataxia, associated debilitation, and the like.

g. Combination of

Can be used alone or in combination with other active substances of the formula 1 according to the inventionA compound of formula 1. The compounds of the general formula 1 can also optionally be combined with other pharmaceutically active substances. These include beta 2-adrenoceptor-agonists (short and long acting), anti-cholinergics (short and long acting), anti-inflammatory steroids (oral and topical corticosteroids), cromoglycate (cromoglycate), methylxanthines, dissociative-glucocorticoid mimetics, PDE3 inhibitors, PDE 4-inhibitors, PDE 7-inhibitors, LTD4 antagonists, EGFR-inhibitors, dopamine agonists, PAF antagonists, lipoxin A4 derivatives, FPRL1 modulators, LTB 4-receptors (BLT1, BLT2) antagonists, histamine H1 receptor antagonists, histamine H4 receptor antagonists, dual histamine H1/H3-receptor antagonists, PI 3-kinase inhibitors, non-receptor tyrosine kinases (such as, for example, LYN, LCK, SYK, ZAP-70, FYN, BTK or ITK), MAP kinases (such as, for example, p38, Inhibitors of ERK1, ERK2, JNK1, JNK2, JNK3 or SAP), inhibitors of the NF-kb signaling pathway (such as, for example, IKK2 kinase inhibitors), iNOS inhibitors, MRP4 inhibitors, leukotriene biosynthesis inhibitors (such as, for example, 5-lipoxygenase (5-LO) inhibitors, cPLA2 inhibitors, leukotriene a4 hydrolase inhibitors or FLAP inhibitors), non-steroidal anti-inflammatory agents (NSAIDs), CRTH2 antagonists, DP 1-receptor modulators, thromboxane receptor antagonists, further CCR3 antagonists, CCR4 antagonists, CCR1 antagonists, CCR5 antagonists, CCR6 antagonists, CCR7 antagonists, CCR8 antagonists, CXCR 9 antagonists, CCR30 antagonists, CXCR3 antagonists, CXCR4 antagonists, CXCR CCR 3554 antagonists ²Antagonists, CXCR1 antagonists, CXCR5 antagonists, CXCR6 antagonists, CX3CR3 antagonists, neurokinins (NK1, NK2) antagonists, sphingosine 1-phosphate receptor modulators, sphingosine 1-phosphate lyase inhibitors, adenosine receptor modulators (such as, for example, A2 a-agonists), modulators of purinic receptors (such as, for example, P2X7 inhibitors), Histone Deacetylase (HDAC) activators, bradykinin (BK1, BK2) antagonists, TACE inhibitors, PPAR γ modulators, Rho-kinase inhibitors, interleukin 1-beta converting enzyme (ICE) inhibitors, Toll-like receptor (TLR) modulators, HMG-CoA reductase inhibitors, VLA-4 antagonists, ICAM-1 inhibitors, SHIP agonists, GABAa receptor antagonists, ENaC-inhibitors, melanocortin receptors (MC1R, MC2R, MC3R, 4R, MC5R) modulators, CGRP antagonists, endothelinsAntagonists, TNF α antagonists, anti-TNF antibodies, anti-GM-CSF antibodies, anti-CD 46 antibodies, anti-IL-1 antibodies, anti-IL-2 antibodies, anti-IL-4 antibodies, anti-IL-5 antibodies, anti-IL-13 antibodies, anti-IL-4/IL-13 antibodies, anti-TSLP antibodies, anti-OX 40 antibodies, mucus regulators, immunotherapeutics, compounds against airway swelling, anti-cough compounds, VEGF inhibitors, and combinations of two or three active agents.

In some embodiments, the additional active agent is betaminometics, anticholinergics, corticosteroids, PDE 4-inhibitors, LTD 4-antagonists, EGFR-inhibitors, CRTH2 inhibitors, 5-LO-inhibitors, histamine receptor antagonists, and SYK-inhibitors, and combinations of two or three active agents, namely:

betamiphene and corticosteroids, PDE 4-inhibitors, CRTH 2-inhibitors or LTD 4-antagonists,

anticholinergics with betaimel, corticosteroids, PDE 4-inhibitors, CRTH 2-inhibitors or LTD 4-antagonists,

corticosteroids with PDE 4-inhibitors, CRTH 2-inhibitors or LTD 4-antagonists

PDE 4-inhibitors with CRTH 2-inhibitors or LTD 4-antagonists

CRTH 2-inhibitors and LTD 4-antagonists.

In these embodiments, the compounds comprising the combination may be administered to the individual in combination. The terms "co-administration" and "in combination with … …" include the simultaneous, concurrent or sequential administration of two or more therapeutic agents without a specific time period. In embodiments, these agents are present within the cell or in the subject at the same time or exert their biological or therapeutic effects at the same time. In one embodiment, the therapeutic agents are in the same composition or unit dosage form. In other embodiments, these therapeutic agents are in separate compositions or unit dosage forms. In certain embodiments, the first agent can be administered prior to (e.g., before, with, or with 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks), concomitantly with, or subsequent to (e.g., after 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks) the administration of the second therapeutic agent. "concomitant administration" of a known therapeutic agent and a pharmaceutical composition of the invention refers to administration of the compound and second agent at a time when both the known agent and the composition of the invention have a therapeutic effect. Such concomitant administration may involve administering the drug simultaneously (i.e., at the same time), before, or after the administration of the compound of interest. The routes of administration of the two agents may differ, with representative routes of administration described in more detail below. One of skill in the art will readily determine the appropriate timing, sequence and dosage for the administration of particular drugs and compounds of the invention. In some embodiments, the compounds (e.g., the subject compound and at least one additional compound) are administered within twenty-four hours of each other, such as within 12 hours of each other, within 6 hours of each other, within 3 hours of each other, or within 1 hour of each other. In certain embodiments, the compounds are administered within 1 hour of each other. In certain embodiments, the compounds are administered substantially simultaneously. By substantially simultaneously administered is meant that the compounds are administered to the individual within about 10 minutes or less, such as 5 minutes or less, or 1 minute or less of each other.

"companion diagnosis" or "companion diagnosis device" refers to an in vitro diagnostic device or imaging means that provides information essential for safe and effective use of a corresponding therapeutic product. The instructions specify the use of the in vitro diagnostic companion device with a particular therapeutic product for labeling the device and the corresponding therapeutic product as well as labeling any universal equivalents and biological analogs of the therapeutic product.

Companion diagnostic tests can take several forms, including by way of example and not limitation: tests to screen for familial genetic patterns and conditions that are difficult to diagnose; a prognostic test to predict future development of a disease; a treatment test that indicates the patient's response to the prescribed therapy; monitoring tests to assess the effectiveness of prescribed therapies and appropriate dosages; and a relapse test to analyze the patient's risk of relapsing disease. See Agarwal A et al, Pharmgenomics Pers Med.8:99-110(2015), which is incorporated herein by reference in its entirety.

h. Pharmaceutical forms

Suitable formulations for administration of the compound of formula 1 and the co-crystals or salt forms of formulae 2 and 2a include, for example, tablets, capsules, suppositories, solutions and powders and the like. The amount of pharmaceutically active compound should be in the range of 0.05 to 90% by weight, such as 0.1 to 50% by weight, of the composition as a whole. Suitable tablets may be obtained, for example, by mixing the active substance with known excipients, for example inert diluents (such as calcium carbonate, calcium phosphate or lactose), disintegrants (such as corn starch or alginic acid), binders (such as starch or gelatin), lubricants (such as magnesium stearate or talc) and/or agents for delaying release (such as carboxymethylcellulose, cellulose acetate phthalate or polyvinyl acetate). The tablet may also include several layers.

Coated lozenges may accordingly be prepared by coating cores produced analogously to lozenges with substances which are customarily used for the coating of lozenges, such as collidone or shellac, gum arabic, talc, titanium dioxide or sugar. The core may also be composed of a number of layers in order to achieve delayed release or to prevent incompatibilities. Similarly, the tablet coating may be composed of a number of layers to achieve delayed release, possibly using excipients as described above for the tablets.

A syrup or elixir containing an active substance or a combination thereof according to the invention may additionally contain a sweetening agent such as saccharin, cyclamate, glycerol or sugar and a flavour enhancer such as a flavour such as vanillin or orange extract. They may also contain suspension adjuvants or thickeners (such as sodium carboxymethylcellulose), wetting agents (such as, for example, condensation products of fatty alcohols with ethylene oxide) or preservatives (such as p-hydroxybenzoates).

Solutions are prepared in a conventional manner, for example with the addition of isotonic agents, preservatives (such as p-hydroxybenzoate) or stabilizers (such as alkali metal salts of ethylenediamine tetraacetic acid), optionally with the use of emulsifiers and/or dispersants, while, for example, if water is used as diluent, organic solvents may optionally be used as solubilizers or dissolution aids, and the solutions may be transferred into injection vials or ampoules or infusion bottles.

Capsules containing one or more than one active substance or combination of active substances may be prepared, for example, by mixing the active substances with inert carriers such as lactose or sorbitol and filling them into gelatin capsules.

Suitable suppositories may be prepared, for example, by mixing with carriers provided for this purpose, such as neutral fats or polyethylene glycols or derivatives thereof.

Excipients that may be used include, for example, water, pharmaceutically acceptable organic solvents such as paraffins (e.g. petroleum fractions), vegetable oils (e.g. peanut or sesame oil), mono-or polyfunctional alcohols (e.g. ethanol or glycerol), carriers such as, for example, natural mineral powders (e.g. kaolin, clay, talc, chalk), synthetic mineral powders (e.g. highly dispersed silicic acid and silicates), sugars (e.g. sucrose, lactose and glucose), emulsifiers (e.g. lignin, spent sulfite liquor, methylcellulose, starch and polyvinylpyrrolidone) and lubricants (e.g. magnesium stearate, talc, stearic acid and sodium lauryl sulfate).

For oral use, the tablets may obviously contain, in addition to the indicated carrier, additives such as sodium citrate, calcium carbonate and dicalcium phosphate, as well as various other substances such as starch (e.g., potato starch), gelatin and the like. Lubricants such as magnesium stearate, sodium lauryl sulfate and talc may also be used to produce tablets. In the case of aqueous suspensions, the active substance may be combined with various flavoring or coloring agents in addition to the excipients mentioned above.

For administration of the compound of formula 1 and the co-crystal or salt form of formulae 2 and 2a, formulations suitable for inhalation or pharmaceutical preparations may be employed. Inhalable formulations include inhalable powders, propellant-containing metered dose aerosols or propellant-free inhalable solutions. Within the scope of the present invention, the term propellant-free inhalable solutions also includes concentrated or sterile inhalable solutions ready for use. The formulations that can be used within the scope of the invention are described in more detail in the next part of the description.

The inhalable powders which can be used according to the invention may contain the compound of formula 1 or the co-crystal or salt form of formulae 2 and 2a, alone or in admixture with suitable physiologically acceptable excipients.

If the active substance of the compound of formula 1 or of the cocrystals or salt forms of formulae 2 and 2a is present in a mixture with a physiologically acceptable excipient, these inhalable powders of the invention can be prepared using the following physiologically acceptable excipients: monosaccharides (e.g. glucose or arabinose), disaccharides (e.g. lactose, sucrose, maltose), oligo-and polysaccharides (e.g. dextrose), polyols (e.g. sorbitol, mannitol, xylitol), salts (e.g. sodium chloride, calcium carbonate) or mixtures of these excipients. In some examples, a mono-or disaccharide, such as lactose or glucose, is used, for example in the form of a hydrate thereof, e.g. lactose, such as lactose monohydrate.

Within the scope of the inhalable powders according to the invention, the excipient has a maximum average particle size of at most 250 μm, such as 10 to 150 μm, and is comprised between 15 and 80 μm. It sometimes seems appropriate to add a finer excipient fraction having an average particle size of 1 to 9 μm to the above excipients. These finer excipients are also selected from the group of possible excipients listed above. Finally, to prepare the inhalable powders of the invention, a micronized active substance, such as a compound of formula 1 or a co-crystal or salt form of formulae 2 and 2a having a mean particle size of 0.5 to 10 μm, including 1 to 5 μm, is added to the excipient mixture. Methods for preparing the inhalable powders according to the invention by grinding and micronization and finally mixing the ingredients together are known in the prior art.

The inhalable powders according to the invention can be administered using inhalers known from the prior art.

The inhalation aerosols of the invention containing a propellant gas may comprise the compound of formula 1 or the co-crystals or salt forms of formulae 2 and 2a dissolved in the propellant gas or in dispersed form. The compounds of formula 1 or the co-crystals or salt forms of formulae 2 and 2a can be contained in separate formulations or in customary formulations, where they are dissolved, dispersed or only one component is dissolved and the other is dispersed in each case. Propellant gases which can be used for the preparation of inhalation aerosols are known from the prior art. Suitable propellant gases are selected from hydrocarbons such as n-propane, n-butane or isobutane, and halogenated hydrocarbons such as methane, ethane, propane, butane, cyclopropane or fluorinated derivatives of cyclobutane. The propellant gases mentioned above may be used individually or in mixtures. In some examples, the propellant gas is selected from halogenated alkane derivatives of TG134a and TG227 and mixtures thereof.

Propellant-driven inhalation aerosols may also contain other ingredients such as cosolvents, stabilizers, surfactants, antioxidants, lubricants, and pH adjusters. All of these ingredients are known in the art.

The propellant-driven inhalation aerosols of the invention described above can be administered using inhalers known in the art (MDI ═ metered dose inhalers).

Furthermore, the active substances according to the invention in the form of the compounds of formula 1 or the co-crystals or salts of formulae 2 and 2a can be applied in the form of propellant-free inhalable solutions and suspensions. The solvent used may be aqueous or alcoholic, such as an ethanol solution. The solvent may be water itself or a mixture of water and ethanol. The relative ratio of ethanol to water is not limited, but in some examples the maximum is at most 70 volume percent, such as at most 60 volume percent and including at most 30 volume percent. The remaining volume consists of water. The solution or suspension comprising the compound of formula 1 or the co-crystal or salt form of formulae 2 and 2a is adjusted to a pH of 2 to 7, such as 2 to 5, using a suitable acid. The pH can be adjusted using an acid selected from inorganic or organic acids. Examples of particularly suitable inorganic acids include hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid and/or phosphoric acid. Examples of particularly suitable organic acids include ascorbic acid, citric acid, malic acid, tartaric acid, maleic acid, succinic acid, fumaric acid, acetic acid, formic acid, and/or propionic acid, and the like. In some examples, the mineral acids are hydrochloric acid and sulfuric acid. It is also possible to use acids which have formed acid addition salts with one of the active substances. In some examples, the organic acids used are ascorbic acid, fumaric acid, and citric acid. Mixtures of the above acids may be used if desired, particularly where the acid has other properties in addition to the acidifying property (e.g. as a fragrance, antioxidant or complexing agent), such as for example citric acid or ascorbic acid. According to the present invention, in some examples, hydrochloric acid is used to adjust the pH.

If desired, the addition of tetraacetic acid (EDTA) or one of its known salts (sodium tetraacetate) as a stabilizer or complexing agent can be omitted from these formulations. Other embodiments may contain this compound or these compounds. In an embodiment, the amount of sodium ethylene glycol tetraacetate based is less than 100mg/100ml, such as less than 50mg/100ml, and including less than 20mg/100 ml. In some examples, an inhalable solution is used in which the sodium tetraacetate is present in an amount of 0 to 10mg/100 ml. Co-solvents and/or other excipients may be added to the propellant-free inhalable solutions, such as those containing hydroxyl groups or other polar groups, for example alcohols-especially isopropanol, glycols-especially propylene glycol, polyethylene glycol, polypropylene glycol, glycol ethers, glycerol, polyoxyethylene alcohols and polyoxyethylene fatty acid esters. The terms excipient and additive herein denote any pharmaceutically acceptable substance which is not an active substance, but which can be formulated together with the active substance in a physiologically suitable solvent in order to improve the qualitative properties of the active substance preparation. In some embodiments, the substance has no pharmacological effect, or is paired with a desired therapy, with no detectable or at least no undesired pharmacological effect. Excipients and additives include, for example, surfactants such as soy lecithin, oleic acid; sorbitol esters, such as polysorbates; polyvinylpyrrolidone; other stabilizers; a complexing agent; antioxidants and/or preservatives that ensure or prolong the shelf life of the finished pharmaceutical formulation; a fragrance; vitamins and/or other additives known in the art. Additives also include pharmaceutically acceptable salts, such as sodium chloride as an isotonic agent.

In some embodiments, the excipients include antioxidants such as ascorbic acid (e.g., with the proviso that it has not been used to adjust pH), vitamin a, vitamin E, tocopherol, and similar vitamins and provitamins found in the human body.

Preservatives can be used to protect the formulation from contamination by pathogens. Suitable preservatives are those known in the art, in particular acetylpyridinium chloride, benzyldimethylammonium chloride or benzoic acid or a benzoate salt, such as sodium benzoate, in concentrations known in the art. The above preservatives may be present at a concentration of up to 50mg/100ml, such as from 5 to 20mg/100 ml.

In some embodiments, the formulation comprises only benzyldimethylammonium chloride and sodium ethylene glycol tetraacetate, in addition to the solvent water and the compound of formula 1 or the co-crystal or salt form of formulae 2 and 2 a. In one embodiment, no sodium tetraacetate oxalate is present.

The dosage of the compounds of the invention is naturally highly dependent on the method of administration and the patient to be treated. When administered by inhalation, the compound of formula 1 or the co-crystal or salt form of formulae 2 and 2a is characterized by high potency even at doses in the μ g range. The compound of formula 1 or the co-crystal or salt form of formulae 2 and 2a may also be used effectively in the μ g range or above. For example, the dosage may be in the gram range.

In a further aspect, the invention relates to the above pharmaceutical preparations per se, characterized in that they contain a compound of formula 1 or a co-crystal or salt form of formulae 2 and 2a, in particular to the above pharmaceutical preparations which can be administered by inhalation.

The following formulation examples illustrate the invention without limiting its scope:

i. examples of pharmaceutical preparations

A)

The finely ground active substance, lactose and some corn starch are mixed together. The mixture was sieved, subsequently moistened with an aqueous solution of polyvinylpyrrolidone, kneaded, wet-granulated and dried. The granules, the remaining corn starch and magnesium stearate are sieved and mixed together. The mixture is compressed into tablets of suitable shape and size.

B)

The finely ground active substance, some corn starch, lactose, microcrystalline cellulose and polyvinylpyrrolidone are mixed together, the mixture is sieved and treated with the remaining corn starch and water to form granules, which are dried and sieved. Sodium carboxymethyl starch and magnesium stearate are added and mixed and the mixture is compressed into tablets of appropriate size.

C)

The active substance is dissolved in water at its own pH or optionally at pH5.5 to 6.5 and sodium chloride is added to make the solution isotonic. The resulting solution was filtered to remove pyrogens and the filtrate was transferred aseptically into ampoules, which were then sterilized and heat sealed. Ampoules contain 5mg, 25mg and 50mg of active substance.

D)

The suspension is transferred to a known aerosol container with a metering valve. Preferably 50. mu.l of suspension is released per actuation. If desired, the active substance may also be released in higher doses (e.g. 0.02% by weight).

E)This solution can be prepared in a usual manner.

F)

Inhalable powders are prepared by mixing the individual ingredients in a conventional manner.

j. Indications of

Methods of improving cognitive or other symptoms of cognitive diseases by treating a subject/patient diagnosed with a cognitive related disease are provided. Aspects of these methods include modulating CCR3, for example using CCR3 modulators, in a manner sufficient to treat a cognitive related disease in a patient. These methods include treating a cognitive related disorder with an orally available and bioavailable composition comprising a compound of formula 1, a co-crystal or salt of formula 2 or 2a, or a formulation of formula 3, as described above. As summarized above and described in more detail below, embodiments of the invention can treat various aging-related injuries, such as cognitive-related diseases. In some examples, the disorder of interest is a cognitive-related disease disorder associated with neurodegeneration, e.g., as evidenced by impaired nerves (such as a reduction in one or more neurogenesis), e.g., as manifested by a reduction in the number of BrdU or EdU-positive cells, Ki 67-positive cells, and Dcx-positive cells when compared to non-diseased tissue. Compositions that modulate CCR3 can be administered to patients/subjects diagnosed with cognitive related diseases such as (by way of example and not limitation): mild Cognitive Impairment (MCI); alzheimer's disease; parkinson's disease; frontotemporal dementia (FTD); huntington's disease; amyotrophic Lateral Sclerosis (ALS); multiple Sclerosis (MS); glaucoma, and glaucoma; myotonic dystrophy; dementia; progressive Supranuclear Palsy (PSP); ataxia; multiple system atrophy; and asthenia (frailty); they are described further below. The methods of the invention may further comprise monitoring the improvement in the progression of neurodegenerative disease by measuring cognitive or physical improvement.

Methods are provided for improving motor coordination, function, or other symptoms by treating a subject/patient diagnosed with a movement disorder. Aspects of these methods include modulating CCR3, for example using CCR3 modulators, in a manner sufficient to treat dyskinesia in a patient. These methods include treating movement disorders with an orally available and bioavailable composition comprising a compound of formula 1, a co-crystal or salt of formula 2 or 2a, or a formulation of formula 3, as described above. As summarized above and described in more detail below, embodiments of the invention can treat a variety of age-related impairments, such as dyskinesias. In some examples, the condition of interest is a movement disorder associated with neurodegeneration, e.g., as evidenced by impaired nerves (such as a reduction in one or more neurogenesis), e.g., as manifested by a reduction in the number of BrdU or EdU positive cells, Ki67 positive cells, and Dcx positive cells when compared to non-diseased tissue. Compositions that modulate CCR3 may be administered to patients/subjects diagnosed with dyskinesias such as (by way of example and not limitation): parkinson's disease; parkinson's disease; dementia with lewy bodies; ataxia; dystonia; cervical dystonia; chorea; huntington's disease, multiple system atrophy; spasm; progressive supranuclear palsy; a bradykinesia; tourette syndrome; and tremor; they are described further below. The methods of the invention may further comprise monitoring the improvement in the progression of neurodegenerative disease by measuring cognitive or physical improvement.

Mild cognitive impairment (m.c.i.) is a mild cognitive disorder that manifests as memory or other mental function (such as planning, following instructions, or making decisions) problems that worsen over time, while overall mental function and daily activities are unaffected. Thus, while significant neuronal death generally does not occur, neurons in the aging brain are susceptible to sub-lethal age-related changes in structure, synaptic integrity, and processing of synaptic molecules, all of which impair cognitive function. Subjects suffering from or at risk of developing an aging-related cognitive disorder (who would benefit from treatment with the subject compounds, e.g., by the methods disclosed herein) also include subjects of any age suffering from a cognitive disorder due to an aging-related disorder; and any age of an individual diagnosed with an aging-related disorder (which is often accompanied by cognitive impairment), wherein the individual has not yet begun to develop symptoms of cognitive impairment. Examples of such aging-related disorders include, in a non-limiting manner, those listed below.

Alzheimer's disease. Alzheimer's disease is characterized by a progressive and inevitable loss of cognitive function associated with an excess of senile plaques in the cerebral cortex and subcortical gray matter and an excess of β -amyloid and neurofibrillary tangles composed of Tau protein. Common forms affect people >60 years of age, and their incidence increases with age. It accounts for more than 65% of senile dementia.

The etiology of Alzheimer's disease is not known. About 15% to 20% of cases of the disease occur in the home. The remaining so-called sporadic cases have some genetic relevance. The disease has a somatotropically dominant inheritance pattern in most cases of early and some late onset, but does not have variable late-stage penetrance. Environmental factors are the focus of active investigation.

During the course of the disease, synapses (and ultimately neurons) are lost in the cerebral cortex, hippocampus, and subcortical structures, including selective cellular loss in the basal nucleus of Meynert (nucleus basalis), locus coeruleus, and the dorsal ventral spine (nucleus dorsallis). Reduced brain glucose use and perfusion in certain areas of the brain (the apical and temporal cortex in early disease and the prefrontal cortex in later disease). Neuritis or senile plaques (consisting of axons, astrocytes and glial cells around amyloid nuclei) and neurofibrillary tangles (consisting of paired helical fibers) play a role in the pathogenesis of alzheimer's disease. Senile plaques and neurofibrillary tangles occur with normal aging, but they are more prevalent in patients with alzheimer's disease.

Parkinson's Disease. Parkinson's Disease (PD) is an idiopathic, slowly progressive degenerative CNS disorder characterized by bradykinesia and reduction (bradykinesia), rigidity of muscles, resting tremor (dystonia), stiffness of muscles and postural instability. PD was originally thought to be primarily dyskinesia, but is now thought to also cause depression and mood changes. PD can also affect cognitive, behavioral, sleep, autonomic and sensory functions. The most common cognitive disorders include impaired attention and concentration, working memory, executive function, production of speech and visual-spatial functions. PD is characterized by symptoms associated with reduced motor function usually preceded by symptoms associated with cognitive impairment, which aids in the diagnosis of the disease.

In primary parkinson's disease, pigmented neurons of the substantia nigra, locus coerulea and other brainstem dopaminergic cell populations degenerate. The reason is unknown. Loss of substantia nigra neurons projecting to the caudate and putamen results in a deficiency of the neurotransmitter dopamine in these regions. The incidence generally increases after age 40 in older age groups.

Approximately 60,000 americans are newly diagnosed with parkinson's disease each year and currently affect approximately one million americans. Although PD itself is not fatal, its complication is the fourteenth leading cause of death in the united states. At present, PD is incurable and treatment is generally used to control symptoms, with surgery being performed in advanced severe cases.

Treatment options for PD include administering drugs to help control motor deficits. These options augment or replace the neurotransmitter dopamine in PD patients with low brain concentrations. Such drugs include: carbidopa (carbidopa)/levodopa (levodopa) (which produces more dopamine in the brain); apomorphine (apomorphine), pramipexole (pramipexole), ropinirole (pramipexole) and rotigotine (rotigotine) (dopamine agonists); selegiline (selegiline) and rasagiline (rasagiline) (MAO-B inhibitors that prevent dopamine decomposition); entacapone (entacapone) and tolcapone (tolcapone) (catechol-O-methyltransferase [ COMT ] inhibitors, which cause more levodopa to be present in the brain); benztropine (benztropine) and trihexyphenidyl (trihexyphenidyl) (anticholinergic); and amantadine (to control tremor and rigidity). Exercise/physical therapy is also commonly used to help maintain physical and mental functions.

However, current treatment options for treating PD symptoms are non-curative and do not prevent disease progression. In addition, current drugs often lose efficacy in late stage PD. The most commonly prescribed drug, levodopa, usually causes adverse effects within 5 to 10 years after the onset of medication. These adverse effects can be severe and can lead to unpredictable fluctuations in motor fluctuations and motor control between doses and spasticity/convulsions (dyskinesias) that are difficult to manage, and even as ineffective as symptoms of PD itself. Thus, there is a need for novel therapies with novel mechanisms of action that can be administered with or in combination with current PD drugs.

Parkinson's disease (Parkinsonism). Secondary Parkinson's disease (also known as atypical Parkinson's disease or Parkinson's plus) results from loss of or interference of dopamine action in the basal ganglia due to other idiopathic degenerative diseases, drugs or exotoxins. The most common cause of secondary parkinson's disease is the ingestion of antipsychotics or reserpine (reserpine), which produces parkinson's disease by blocking dopamine receptors. Uncommon causes include carbon monoxide or manganese poisoning, hydrocephalus, structural lesions (tumors, infarcts affecting the midbrain or basal ganglia), subdural hematomas, and degenerative disorders (including degeneration of the nigrostriatal body). Certain diseases, such as Progressive Supranuclear Palsy (PSP), Multiple System Atrophy (MSA), corticobasal degeneration (CBD), and dementia with lewy bodies (DLB), may manifest parkinsonian symptoms prior to the primary symptoms necessary to make a particular diagnosis, and thus may be labeled as "parkinson's disease".

Assessment of the progression of PD

Several rating scales have been used to assess the progress of PD. The most widely used scales include the Unified Parkinson's Disease Rating Scale (UPDRS, proposed in 1987) (j.rehibil res. dev., 201249 (8):1269-76) and the Hoehn and Yahr Scale (neurology, 196717 (5): 427-42). Other scales include the dyskinesia association (MDS) updated UPDRS scale (MDS-UPDRS) and the Schwab and England Activities of Daily Living (ADL) scale.

The UPDRS scale evaluates 31 items, which constitute three sub-scales: (1) mental, behavioral and emotional; (2) activities of daily living; and (3) motion checking. The Hoehn and Yahr scale divides PD into five stages with discreet sub-stages: 0-no signs of disease; 1-symptoms appear on one side only; 1.5-lateral symptoms, but also related to the neck and spine; 2-bilateral symptoms, no balance disorder; 2.5-mild bilateral symptoms, recovery when a "pull" test is provided; 3-dysbalance in mild to moderate disease; 4-severely disabled, but able to walk or stand independently; and 5-need wheelchair or bed rest without help. The Schwab and England scale classifies PD as a number of percentages (from 100% -completely independent to 10% -completely dependent). Frontotemporal dementia. Frontotemporal dementia (FTD) is a disease caused by progressive degeneration of the frontal lobe of the brain. Over time, degeneration may progress to the temporal lobe. FTD accounts for 20% of the cases of pre-senile dementia, second only to the prevalence of Alzheimer's Disease (AD). Symptoms were divided into three groups based on the function of the affected frontal and temporal lobes:

Behavioral modification type ftd (bvftd), symptoms of which include manifestation of sleepiness and involuntary nature on the one hand, and release of inhibition on the other hand; progressive non-fluent aphasia (PNFA), in which reduced fluency of speech due to pronunciation difficulties, phonetic and/or syntactic errors is observed, but lexical understanding is preserved; and Semantic Dementia (SD), where patients maintain fluent normal speech and syntax, but are increasingly difficult in naming and lexical understanding. Other cognitive symptoms common to all FTD patients include impaired executive function and attention. Other cognitive abilities (including perception, spatial skills, memory, and practice) generally remain intact. FTD can be diagnosed by observing frontal and/or anterior temporal lobe atrophy in a structural MRI scan.

There are many forms of FTD, any of which can be treated or prevented using the subject methods and compositions. For example, one form of frontotemporal dementia is Semantic Dementia (SD). SD is characterized by loss of semantic memory in both verbal and nonverbal domains. Patients with SD often suffer from difficulties in finding words. Clinical signs include fluent aphasia, inability to name, impaired lexical meaning comprehension, and associative visual insights (inability to match semantically related pictures or objects). As the disease progresses, behavior and personality changes are often found similar to those found in frontotemporal dementia, although cases are described as "pure" semantic dementia with few symptoms of later-stage behavior. Structural MRI images show a characteristic map of temporal lobe atrophy (mainly on the left), with lower progression greater than upper, and anterior temporal lobe atrophy greater than posterior.

As another example, another form of frontotemporal dementia is Pick's disease (PiD, also PcD). A key feature of the disease is the accumulation of tau protein in neurons, accumulating into silver-stained spherical aggregates known as "Pick bones". Symptoms include loss of speech (aphasia) and dementia. Patients with orbital-frontal dysfunction may become aggressive and socially inappropriate.

They may steal or show compulsive or repetitive improvisation. Patients with medial or lateral dorsolateral frontal lobe dysfunction may exhibit a lack of attention, apathy, or spontaneous decline. Patients may exhibit loss of self-monitoring, abnormal self-consciousness, and an inability to understand significance.

Patients who lose gray matter in the bilateral posterolateral orbital-frontal cortex and right anterior cerebral island may exhibit changes in eating behavior, such as pathological sweet taste. Patients who lose more focal gray matter in the anterolateral orbital frontal cortex may develop hyperphagia. While some symptoms may initially be alleviated, the disease progresses and patients typically die within two to ten years.

Huntington's disease. Huntington's Disease (HD) is a genetically progressive neurodegenerative disease characterized by abnormalities in the development of mood, behavior and spirit; loss of intellectual or cognitive function; and dyskinesias (dyskinesias). Typical symptoms of HD include the development of chorea, which can affect involuntary, rapid, irregular, reflex movements of the face, arms, legs or trunk, as well as cognitive decline, including the gradual loss of thought process and acquired mental capacity. There may be damage to memory, abstract thinking and judgment; misperception of time, place, or identity (disorientation); increased anxiety disorder; and personality changes (personality splitting). Although symptoms typically become apparent during the fortieth or fiftieth years of life, the age of onset can vary and range from infancy to advanced adulthood (e.g., over 70 or over 80 years).

HD spreads in a somatic dominant trait in the home. This condition occurs due to an abnormally long sequence or "repeat" of the coding instructions within the gene on chromosome 4 (4p 16.3). The progressive loss of nervous system function associated with HD results from the loss of neurons in certain areas of the brain, including the basal ganglia and the cerebral cortex.

Amyotrophic lateral sclerosis. Amyotrophic Lateral Sclerosis (ALS) is a rapidly progressive, invariably fatal neurological disease that attacks motor neurons. It was initially noted that symptoms of muscle weakness and atrophy, as well as anterior horn cell dysfunction, are most common in the hands and less common in the feet. The sites of disease are random and progress asymmetrically. Cramps are common and can precede debilitation. Few patients survive for 30 years; 50% die within 3 years of morbidity, 20% survive 5 years, and 10% survive 10 years.

Diagnostic features include onset in the middle or late stages of adult life and progressive motor involvement without paresthesia. Nerve conduction velocity is normal until late stage of disease. Recent studies have also documented the manifestation of cognitive disorders, particularly the reduction of real-time speech memory, visual memory, speech and executive function.

Reductions in cell body area, number of synapses and total synapse length have been reported even in normally occurring neurons in ALS patients. It has been shown that when plasticity of the active region reaches a limit, the continued loss of synapses may lead to functional impairment. Promoting the formation of new synapses or preventing loss of synapses may maintain neuronal function in these patients.

Multiple sclerosis. Multiple Sclerosis (MS) is characterized by various symptoms and signs of CNS dysfunction, which are remitted and aggravated relapses. The most commonly occurring symptoms are paresthesias in one or more limbs, trunk or one side of the face; weakness or awkwardness of the legs or hands; or a visual disorder, such as partial blindness and pain in one eye (retrobulbar neuritis), blurred vision, or vertigo. Common cognitive disorders include impairment of memory (acquiring, retaining and retrieving new information), attention and concentration (particularly distractions), information processing, executive function, visual-spatial function and verbal fluency. Common early symptoms are paralysis of the eyes (which leads to double vision (double vision)), transient weakness of one or more limbs, slight stiffness or abnormal susceptibility to fatigue of the limbs, mild gait disturbances, difficulty in bladder control, dizziness, and mild mood disturbances; they all indicate decentralized CNS involvement and usually confirm that the disease occurred months or years before. Overheating can exacerbate symptoms and signs.

The process is very different, unpredictable, and (in most patients) intermittent. Initially, remissions of months or years can occur separately, particularly when the disease begins with retrobulbar neuritis. However, some patients often develop and rapidly lose energy; a few processes can progress rapidly.

Glaucoma is caused. Glaucoma is a common neurodegenerative disease affecting Retinal Ganglion Cells (RGCs). Evidence supports the existence of separate degeneration programs in synapses and dendrites (including in RGCs). Recent evidence also indicates a correlation between cognitive impairment in elderly and glaucoma (Yochim BP et al, Presence of cognitive impairment, expression, and inertia systems amplitude masses with glaucoma. J Glaucoma. 2012; 21(4): 250-.

Myotonic dystrophy. Myotonic Dystrophy (DM) is a somatomatotonic multisystemic condition characterized by dystrophic muscle weakness and myotonia. The molecular defect is an amplified trinucleotide (CTG) repeat in the 3' untranslated region of the actin protein kinase gene on chromosome 19 q. Symptoms can occur at any age and range of clinical severity. Myotonia is prominent in hand muscles, and ptosis is common even in mild cases. In severe cases, there is significant peripheral muscle weakness, often accompanied by cataracts, premature baldness, the hatchling phase, cardiac arrhythmias, testicular atrophy, and endocrine abnormalities (e.g., diabetes). Mental retardation is common in severe congenital forms, while decline in cognitive function (particularly speech and executive function) of the frontal and temporal lobes associated with aging is observed in the milder adult form of the disorder. Severely affected persons die at the age of 50.

Dementia. Dementia describes a group of disorders with symptoms that affect thinking and social abilities sufficiently severe to interfere with daily functioning. In addition to the dementia observed in the late stage of the aging-related disorder discussed above, other examples of dementia include vascular dementia and dementia with lewy bodies as described below.

In vascular dementia or "multi-infarct dementia," cognitive impairment is caused by a problem with blood supply to the brain, typically by a series of mild strokes, or sometimes a large stroke before or after other smaller strokes. The vascular lesions may be the result of diffuse cerebrovascular disease, such as small vessel disease or localized lesions or both. Patients suffering from vascular dementia develop cognitive disorders acutely or subacute after an acute cerebrovascular event, after which progressive cognitive decline is observed. Cognitive disorders are those similar to those observed in alzheimer's disease, including speech, memory, complex visual processing or executive dysfunction, although the associated changes in the brain are not due to AD pathology, but rather due to chronic reduced blood flow in the brain, ultimately resulting in dementia. Single Photon Emission Computed Tomography (SPECT) and Positron Emission Tomography (PET) neuroimaging can be used to confirm the diagnosis of multi-infarct dementia and assessments related to mental status examinations.

Dementia with lewy bodies.

Dementia with lewy bodies (DLB), also known by various other names including lewy body dementia, diffuse lewy body disease, cortical lewy body disease and senile dementia of the lewy type, is a type of dementia characterized anatomically by the presence of lewy bodies (alpha-synuclein and ubiquitin protein clots) in neurons, detectable in postmortem brain histology. It is mainly characterized by cognitive (especially executive function) decline. Alertness and short-term memory will rise and fall.

Permanent or recurrent visual hallucinations with vivid and detailed pictures are often early diagnostic symptoms. DLB is often confused with alzheimer's disease and/or vascular dementia in its early stages, although, where alzheimer's disease usually begins quite slowly, DLB usually has a rapid or acute onset. DLB symptoms also include motor symptoms similar to those of parkinson's disease. DLB is distinguished from dementia that sometimes occurs in parkinson's disease by the time frame in which dementia symptoms appear relative to parkinson's symptoms. Parkinson's disease (PDD) with dementia will be diagnosed when onset of dementia-type Parkinson's disease occurs more than one year later. DLB is diagnosed when cognitive symptoms begin at the same time or within a year as parkinson's symptoms.

Treatment of DLB is a complex process and requires multiple approaches. (Neurology,201789: 1-13). Typical parkinson's disease therapies such as dopaminergic and anticholinergic drugs can exacerbate cognitive and behavioral symptoms. Optimal treatment typically utilizes both pharmacological (motor, cognitive training and caregiver meta-directed training) and non-pharmacological approaches. For cognitive symptoms, acetylcholinesterase inhibitors (e.g., rivastigmine, donepezil) may be administered, as well as NMDA receptor antagonists (memantine). For neuropsychiatric symptoms, acetylcholinesterase inhibitors improve apathy and hallucinations. Unfortunately, antipsychotics increase the risk of death in DLB patients. In DLB patients, motor symptoms respond poorly to dopaminergic treatment and can exacerbate the risk of psychosis. Levodopa can be used, but only at low threshold doses, and there is therefore a clear need in the field of novel agents for the treatment of DLB.

Progressive supranuclear palsy. Progressive Supranuclear Palsy (PSP) is a brain disorder that causes serious and progressive problems in gait and balance control, as well as complex eye movement and thinking problems. One of the typical symptoms of the disease is the inability to properly target the eye, which occurs due to foci in the brain region that coordinate eye movements. Some individuals describe this effect as fuzzy. Affected individuals often exhibit mood and behavior changes, including depression and apathy, as well as progressive mild dementia. The long name for the disease indicates that the disease slowly begins and continues to worsen (progressive) and causes weakness (paralysis) by damaging specific parts of the brain that control eye movement (supranuclear) above pea-sized structures called the nucleus. PSP was first described as a distinct disease in 1964, when three scientists published a paper that distinguished this condition from parkinson's disease. Sometimes referred to as the Steele-Richardson-olsnowski syndrome, reflects the combined name of the scientists defining the condition. While PSPs progressively get worse, no one dies of the PSPs themselves.

Ataxia is caused by ataxia. People with ataxia have problems with coordination because the part of the nervous system that controls movement and balance is affected. Ataxia can affect the movements of the fingers, palm, arms, legs, trunk, voice and eyes. Word-group ataxia is often used to describe uncoordinated symptoms that may be associated with infection, injury, other diseases, or degenerative changes in the central nervous system. Ataxia is also used to represent a group of specific degenerative diseases of the nervous system called genetic and sporadic Ataxia, which is the major focus of the National Ataxia Foundation.

Multiple system atrophy. Multiple System Atrophy (MSA) is a degenerative neurological disease. MSA is associated with the degeneration of nerve cells in specific areas of the brain. This cellular degeneration causes problems with movement, balance, and other autonomic functions of the body, such as bladder control or blood pressure regulation.

The cause of MSA is unknown and no specific risk factors are found. Approximately 55% of cases occur in men, with the typical age of onset ending at 50 years to the beginning of 60 years. MSA often shows some of the same symptoms as parkinson's disease. However, MSA patients typically show minimal (if any) response to dopamine drugs used in parkinson's disease.

Dystonia. Dystonia is a condition involving sustained involuntary muscle contraction. Such contractions may manifest themselves as a twisting repetitive motion. This condition can affect the entire body or specific parts of the body, each known as a systemic dystonia or a local dystonia. Cervical dystonia can lead to persistent or intermittent contractions of the neck muscles. Dystonia cannot be cured. Current treatment methods include carbidopa-levodopa, benidil, benztropine, tetrabenazine (tetrabenazine), diazepam (diazepam), clonazepam (clonazepam), baclofen (baclofen), physiotherapy, speech therapy, stretching, massage and invasive surgery.

And (5) weakening. Debilitating syndrome ("debilitation") is an aging syndrome characterized by functional and physical decline, including hypomotility, muscle weakness, physical slowness, poor endurance, low physical activity, malnutrition, and involuntary weight loss. These decline are often associated with and a consequence of diseases such as cognitive dysfunction and cancer. However, even without disease, debilitation can occur. Individuals with debilitating illnesses have an increased negative prognostic risk due to fractures, accidental falls, disabilities, complications, and premature death. (C.Buigues et al, Effect of a predictive Formulation on framework Syndrome: ARandomized, Double-blade Clinical Trial, int.J.mol.Sci.2016,17,932). In addition, the incidence of higher healthcare expenditures in individuals suffering from debilitation is increasing. (same as above)

Common symptoms of debilitation can be determined by a particular type of test. For example, unintended weight loss involves losing at least 10 pounds or more than 5% of the body weight of the previous year; muscle weakness can be determined by a minimum of 20% of the baseline decrease in grip strength (adjusted for gender and BMI); body slowness may be based on the time required to walk a 15 foot distance; poor tolerance can be determined by an individual's self-report of fatigue; and low physical activity can be measured using a standardized questionnaire. (Z.Palace et al, The framework Syndrome, Today's Geriatric Medicine 7(1), at 18 (2014)).

In some embodiments, the targeted methods and compositions are applied to slow the progression of cognitive, motor, or other age-related impairments associated with aging. In other words, the cognitive, motor, or other abilities of an individual decrease more slowly after treatment by the disclosed methods than before or without treatment by the disclosed methods. In some such examples, the targeted treatment method comprises measuring the progression of cognitive, motor, or other age-related decline after treatment and determining a decrease in progression of the decline. In some such examples, the determination is made by comparison to a reference, e.g., a rate of decline of the individual prior to treatment, e.g., as determined by measuring cognitive, motor, or other age-related abilities prior to two or more time points prior to administration of the individual's blood product.

The subject methods and compositions are also applied to stabilize cognitive, motor, or other abilities in a subject (e.g., a subject suffering from or at risk of suffering from aging-related cognitive decline). For example, an individual may exhibit some aging-related cognitive impairment, and the progression of cognitive impairment observed prior to treatment using the disclosed methods will cease after treatment with the disclosed methods. As another example, an individual may be at risk of developing an aging-related cognitive decline (e.g., the individual may be 50 years of age or older, or may be diagnosed with an aging-related disorder), and the individual's cognitive ability is substantially unchanged after treatment with the disclosed method as compared to prior to treatment with the disclosed method, i.e., no cognitive decline may be detected.

The subject methods and compositions are also applied to alleviate cognitive, motor, or other age-related impairments in a subject suffering from aging-related impairments. In other words, the affected capacity of the individual is improved after treatment with the target method. For example, the cognitive ability of an individual is increased after treatment with the target method by, e.g., 2-fold or more, 5-fold or more, 10-fold or more, 15-fold or more, 20-fold or more, 30-fold or more or 40-fold or more, including 50-fold or more, 60-fold or more, 70-fold or more, 80-fold or more, 90-fold or more or 100-fold or more, relative to the cognitive ability observed in the individual prior to treatment with the target method.

In some examples, treatment with the subject methods and compositions restores cognitive, motor, or other abilities of an individual suffering from age-related cognitive or motor decline to a level, for example, when the individual is about 40 years of age or less. In other words, cognitive or motor impairment is eliminated.

k. Methods of diagnosing and monitoring improvements in neurodegenerative-related diseases

One of ordinary skill will recognize that in a variety of methods of diagnosing and monitoring disease progression and amelioration of neurodegenerative-related diseases, the following types of assessments may be used alone or in combination with individuals suffering from neurodegenerative diseases. The following types of methods are presented as examples and are not limited to the methods. One of ordinary skill will recognize that other methods of monitoring disease will be useful in practicing the present invention. Those methods may also be encompassed by the present methods.

i. General cognition

The methods of the invention also include methods of monitoring the effectiveness of a drug or treatment for treating cognitive disorders and/or age-related dementia in an individual, comprising comparing cognitive function before and after the treatment. One of ordinary skill will recognize that there are well known methods of assessing cognitive function. For example, and not by way of limitation, the method can include assessing cognitive function based on medical history, family history, physical and neurological examinations by clinicians specializing in treating dementia and cognitive function, laboratory tests, and neuropsychological assessments. Other embodiments encompassed by the present invention include: assessment of consciousness, such as using the Glasgow Coma Scale (Glasgow Coma Scale) (EMV); mental state examination, including simplified mental test scoring (AMTS) or careful mental state examination (MMSE) (Folstein et al, J.Psychiator. Res 1975; 12: 1289-; overall evaluation for higher functionality; such as assessing intracranial pressure by ophthalmoscopy.

In one embodiment, examination of the peripheral nervous system can be used to assess cognitive function, including any of the following: olfactory, visual field and acuity, eye movement and pupils (sympathetic and parasympathetic), facial sensory function, facial and shoulder belt muscle strength, hearing, taste, pharyngeal movement and reflexes, tongue movement, which can be tested individually (e.g., visual sensitivity can be tested by Snellen chart; reflex hammer for testing reflexes including masseter, biceps brachii and triceps tendon, knee tendon, ankle and sole (i.e., Babinski symptoms); muscle strength is typically 1 to 5 on MRC scale; muscle tone and stiffness symptoms).

Multiple sclerosis

In addition to monitoring improvement in cognition-related symptoms, techniques well known to those of ordinary skill can be used to monitor the progression or improvement of neurodegeneration associated with Multiple Sclerosis (MS). By way of example and not limitation, monitoring may be by techniques such as: cerebrospinal fluid (CSF) monitoring; magnetic Resonance Imaging (MRI) to detect the development of lesions and demyelinating plaques; evoked potential studies; and gait monitoring.

CSF analysis can be performed, for example, by lumbar puncture to obtain pressure, appearance and CSF content. Normal values are typically in the following ranges: pressure (70 to 180mm H ₂O); the appearance is clear and colorless; total protein (15 to 60mg/100 mL); IgG is 3 to 12% of total protein; glucose is 50 to 80mg/100 mL; cell count is 0 to 5 white blood cells and no red blood cells; chloride (110 to 125 mEq/L). An abnormal result may indicate the presence or progression of an MS.

MRI is another technique that can be used to monitor disease progression and improvement. Typical criteria in monitoring MS with MRI include the appearance of abnormal white matter patch-like areas in the hemispheres and paraventricular regions of the brain, lesions present in the cerebellum and/or brainstem, and in the cervical or thoracic regions of the spinal cord.

Evoked potentials can be used to monitor the progression and amelioration of MS in an individual. Evoked potential measures such as relaxation of electrical impulses in the Visual Evoked Response (VER), Brainstem Auditory Evoked Response (BAER), and somatosensory evoked response (SSER). Abnormal responses help indicate a decrease in conduction velocity in the central sensory pathway.

Gait monitoring can also be used to monitor disease progression and improvement in MS individuals. MS is often associated with impaired mobility and gait abnormalities, due in part to fatigue. For example, monitoring may be performed using a mobile monitoring device worn by the individual. (Moon, Y., et al, Monitoring gain in multiple sclerasis with novel wearable motion sensors, PLOS One,12(2): e0171346 (2017)).

Huntington's disease

In addition to monitoring improvement in cognition-related symptoms, techniques well known to those of ordinary skill can be used to monitor the progression or improvement of neurodegeneration associated with Huntington's Disease (HD). By way of example and not limitation, monitoring may be by techniques such as: a motion function; a behavior; evaluating the function; and imaging.

Examples of motor functions that may be monitored as an indication of disease progression or improvement include chorea and dystonia, rigidity, bradykinesia, oculomotor dysfunction, and gait/balance changes. Techniques for performing monitoring of these metrics are well known to those of ordinary skill. (see Tang C et al, Monitoring Huntington's disease progression through preclinical and early stages, neurogene Dis Manag 2(4):421-35 (2012)).

The psychotic effects of HD provide an opportunity to monitor disease progression and improvement. For example, a psychiatric diagnosis may be made to determine whether an individual suffers from depression, irritability, anxiety, apathy, and psychosis associated with paranoia. (same as above)

Functional assessment can also be used to monitor disease progression or improvement. Total functional scoring techniques have been reported (supra) and generally decline a little per year in some HD groups.

MRI or PET may also be used to monitor disease progression or improvement. For example, there is loss of striatal projection neurons in HD, and changes in the number of these neurons can be monitored in an individual. Techniques for determining neuronal changes in HD individuals include imaging dopamine D₂Receptor binding. (same as above)

iv.ALS

In addition to monitoring improvement in cognition-related symptoms, techniques well known to those of ordinary skill can be used to monitor the progression or improvement of neurodegeneration associated with Amyotrophic Lateral Sclerosis (ALS). By way of example and not limitation, monitoring may be by techniques such as: evaluating the function; measuring muscle strength; measuring respiratory function; measuring Lower Motor Neuron (LMN) loss; and measuring Upper Motor Neuron (UMN) dysfunction.

Functional assessment can be performed using a functional scale known to those of ordinary skill, such as the ALS functional rating scale (ALSFRS-R), which assesses symptoms associated with medullary, limb, and respiratory function. The rate of change is useful for predicting survival as well as disease progression or improvement. Another measure includes joint assessment of function and survival (CAFS), ranking the clinical outcome of an individual by combining time to live with changes in ALSFRS-R. (Simon NG et al, quantitative Disease progress in Amyotropic lactic acid Sclerosis, Ann neuron 76:643-57 (2014)).

Muscle strength can be tested and quantified by using a composite Manual Muscle Test (MMT) score. This requires average measurements taken from multiple muscle groups using the Medical Research Council (MRC) muscle strength rating scale. Hand-held dynamometer (HHD), among other techniques, may also be used (as above). (same as above)

Respiratory function may be performed using a portable spirometer for obtaining a baseline Forced Vital Capacity (FVC) to predict progression or improvement of disease. In addition, maximum inspiratory pressure, Sniffing Nasal Inspiratory Pressure (SNIP), and small mouth fvc (filling fvc) can be determined and used to monitor disease progression/improvement. (same as above)

Loss of lower motor neurons can be used to monitor the progression or another measure of improvement in ALS disease. Neurophysiologic indices may be determined by measuring the Complex Muscle Action Potential (CMAP) in motor nerve conduction studies, where the parameters include CMAP amplitude and F-wave frequency. (see de Carvalho M et al, supra, New control students in amyotrophic cellular research. Muscle New 23: 344-352, (2000)). The number of lower motor neuron units (MUNE) can also be estimated. In MUNE, the number of residual motor axons that contribute to the maximal CMAP response by estimating individual motor units is estimated and used to determine disease progression or improvement. (Simon NG et al, supra). Other techniques for determining loss of LMN include testing for neural excitability, electrical impedance electromyographs, and detecting changes in muscle thickness using muscle ultrasound. (same as above)

Dysfunction of upper motor neurons can be used to monitor the progression or another measure of improvement in ALS disease. Techniques for determining dysfunction include MRI or PET scanning of the brain and spinal cord, transcranial magnetic stimulation; and determining the level of the biomarker in cerebrospinal fluid (CSF).

Glaucoma, v. glaucoma

In addition to monitoring improvement in cognition-related symptoms, techniques well known to those of ordinary skill can be used to monitor the progression or improvement of the neurodegeneration associated with glaucoma. By way of example and not limitation, monitoring may be by techniques such as: measuring intraocular pressure; assessing damage to the optic dish or optic nerve head; visual field testing for peripheral vision loss; and imaging the dish and retina for topography analysis.

Progressive supranuclear neuroparalgia (PSP)

In addition to monitoring improvement in cognition-related symptoms, techniques well known to those of ordinary skill can be used to monitor the progression or improvement of neurodegeneration associated with Progressive Supranuclear Palsy (PSP). By way of example and not limitation, monitoring may be by techniques such as: functional assessment (activities of daily living, or ADL); evaluating the movement; determination of psychiatric symptoms; and volumetric and functional Magnetic Resonance Imaging (MRI).

In the case of independence, partial dependence on others, or complete dependence, the functional level of an individual may be useful in determining the progression or amelioration of a disease. (see Duff, K et al, Functional impact in progressive subclauar palsy, Neurology 80:380-84, (2013)). The progressive supranuclear neuroparalgia rating scale (PSPRS) is a rating scale that includes twenty-eight measures in six categories: daily activities (by history); a behavior; medulla oblongata, eye movement, limb movement, and gait/midline. The results were a fraction of 0 to 100. Six scores of 0 to 2, twenty-two scores of 0 to 4, and perhaps a total of 100PSPRS scores are practical measurement indicators and are powerful patient survival prediction indicators. They are also sensitive to disease progression and are useful for monitoring disease progression or improvement. (Golbe LI et al, A clinical rating scale for progressive subclauar palsy, Brain 130:1552-65, (2007)).

The ADL portion from UPDRS (unified parkinson's disease rating scale) can also be used to quantify functional activity in individuals with PSP. (Duff K et al, supra). Similarly, Schwab and England Activity daily Life score (SE-ADL) can be used to assess independence. (supra) furthermore, the motor function part of UPDRS is useful as a reliable measure to assess disease progression in PSP patients. For example, the motion part may contain measurements of the motor function of, for example, 27 different quantitative PSP patients. Examples of these include resting tremor, rigidity, finger taps, posture and gait). Disease progression or improvement in an individual can also be assessed by conducting a baseline neuropsychological assessment performed by trained medical personnel using the neuropsychological scale (NPI) to determine the frequency and severity of behavioral abnormalities such as delusions, hallucinations, anxiousness, depression, anxiety, euphoria, lethargy, disinhibition, irritability, and abnormal motor behavior. (same as above)

Functional mri (fmri) may also be used to monitor disease progression and improvement. fMRI is a technique that uses MRI to measure changes in brain activity in certain areas of the brain, typically based on blood flow to these areas. Blood flow is thought to be associated with brain region activation. Patients with neurodegenerative diseases such as PSP may receive physical or mental tests prior to or during scanning in the MRI scanner. By way of example and not limitation, a test may be a sophisticated force control paradigm in which the patient is asked to generate force with the hand most affected by PSP and the Maximum Voluntary Contraction (MVC) is measured by fMRI immediately after the test occurs. (Burciu, RG et al, Distingction patterns of diagnostic in reactive delivery patterns and Parkinson's disease, Mov. Disord.30(9):1248-58 (2015)).

Volumetric MRI is a technique in which an MRI scanner determines volumetric differences in regional brain volumes. This can be done, for example, by comparing different conditions or by measuring the volume difference of the brain regions of a patient over time. Volumetric MRI can be used to determine disease progression or improvement of neurodegenerative disorders such as PSP. This technique is well known to the skilled person. (Messina D et al, Pattern of mail address in Parkinson's disease, progressive subclauar page and multiple system address, Parkinsonism and Related Disorders,17(3):172-76 (2011)). Examples of measurable brain regions include, but are not limited to, intracranial volume, cerebral cortex, cerebellar cortex, thalamus, caudate nucleus, putamen, globus pallidus, hippocampus, amygdala, lateral ventricle, third ventricle, fourth ventricle, and brainstem.

Neurogenesis

Noninvasive techniques for assessing neurogenesis have been reported. (Tamura Y et al, J.Neurosci. (2016)36(31): 8123-31). With tracer [ 2 ]¹⁸F]Positron Emission Tomography (PET) used with FLT, in combination with the BBB transporter inhibitor probenecid, allows tracer accumulation in the neurogenic region of the brain. This imaging allows assessment of neurogenesis in patients receiving treatment for neurodegenerative diseases.

Parkinson's disease and motor function

Several rating scales have been used to assess the progress of PD. The most widely used scales include the Unified Parkinson's Disease Rating Scale (UPDRS, proposed in 1987) (j. rehabil res. dev., 201249 (8):1269-76) and the Hoehn and Yahr Scale (Neurology, 196717 (5): 427-42). Other scales include the dyskinesia association (MDS) updated UPDRS scale (MDS-UPDRS) and the Schwab and England activities of daily living scale (ADL) scale.

The UPDRS scale evaluates 31 items, which constitute three sub-scales: (1) mental, behavioral and emotional; (2) activities of daily living; and (3) motion checking. The Hoehn and Yahr scale divides PD into five stages with discreet sub-stages: 0-no signs of disease; 1-symptoms appear on one side only; 1.5-lateral symptoms, but also related to the neck and spine; 2-bilateral symptoms, no balance disorder; 2.5-mild bilateral symptoms, recovery when a "pull" test is provided; 3-dysbalance in mild to moderate disease; 4-severely disabled, but able to walk or stand independently; and 5-need wheelchair or bed rest without help. The Schwab and England scale divides PD into several percentages (from 100% -completely independent to 10% -completely dependent).

General athletic function may be evaluated using widely used scales, including the general athletic function scale (GMF). This test consists of three components: dependence, pain and restlessness. (Aberg A.C. et al, (2003) Disabll. Rehabil.2003, 5.6.month, 25(9): 462-72.). Athletic performance may also be evaluated using home monitoring or wearable sensors. For example: gait (speed of movement, variability, leg rigidity) can be sensed with an accelerometer; sensing a gesture (torso tilt) by a gyroscope; sensing leg movement by an accelerometer; sensing hand motion by an accelerometer and a gyroscope; sensing tremor (amplitude, frequency, duration, asymmetry) by an accelerometer; sensing a fall by an accelerometer; sensing gait freeze by an accelerometer; sensing dyskinesia through an accelerometer, a gyroscope and an inertial sensor; bradykinesia (duration and frequency) was observed by accelerometer plus gyroscope, and aphasia (pitch) was sensed using a microphone. (Pasorino M et al, Journal of Physics: Conference Series 450(2013) 012055).

Reagent, device and kit

Reagents, devices and kits for carrying out one or more of the above methods are also provided. The target reagents, devices, and kits thereof can vary widely. Agents and devices of interest include those described above for methods of administering a compound of formula 1 in a subject.

In addition to the components described above, the target kit will also include instructions for practicing the target method. These instructions may be present in the target kit in various forms, one or more of which may be present in the kit. One form in which these instructions may be present is in the form of printed information on a suitable medium or substrate, e.g., one or more sheets of paper on which the information is printed, in the packaging of the kit, in package instructions, etc. Alternatively, a computer readable medium, such as a diskette, a CD, a portable flash drive, etc., has information recorded thereon. Another possible way is a website address where information can be accessed at a remote site via the internet. Any convenient means may be present in the kit.

Examples VII. examples

The following examples are provided by way of illustration and not limitation.

a. Pharmaceutical preparation

Pharmaceutical compositions comprising the above compounds, co-crystals and salts can be synthesized, prepared and formulated using the examples disclosed in U.S. patent application publication nos. 2013/0266646, 2016/0081998, U.S. patent nos. 8,278,302, 8,653,075, RE 45323, 8,742,115, 9,233,950 and 8,680,280, which are incorporated herein by reference in their entirety, for administration to an individual suffering from a cognitive or neurodegenerative disease. In addition, these pharmaceutical compositions can be prepared as described in the examples below:

1. Lozenge formulation-Wet granulation

Copovidone was dissolved in ethanol at ambient temperature to produce a granulation liquid. The active CCR3 antagonist ingredient, lactose, and partially cross-linked povidone were blended in a suitable mixer to create a pre-blend. The premix is wetted with a granulation liquid and subsequently granulated. The wet granulate is optionally sieved through a sieve having a mesh size of 1.6 to 3.0 mm. The granules were dried in a suitable dryer at 45 ℃ to a residual moisture content corresponding to a loss on drying of 1 to 3%. The dried granules were sieved through a sieve with a mesh size of 1.0 mm. The granules are blended with a portion of crospovidone and microcrystalline cellulose in a suitable mixer. Magnesium stearate was added to this blend after the lumps were removed by a 1.0mm sieve. The final blend is then produced by final blending in a suitable mixer and compressed into lozenges. The following lozenge compositions were obtained:

components	mg/lozenge	A tablet
			Active ingredient	28.500	30.0
Cross-linked polyvidone	1.500	1.6
			Lactose	28.000	29.5
Co-polyvidone	3.000	3.2
			In total (granules)	61.000	64.3
Microcrystalline cellulose	31.000	32.6
			Cross-linked polyvidone	2.500	2.6
Magnesium stearate	0.500	0.5
			Total of	95.000	100.000

2. Lozenge formulation-melt granulation

The active CCR3 antagonist ingredient, lactose, part mcc, polyethylene glycol, lactose, and part crospovidone were mixed in a suitable mixer to create a premix. The premix is heated in a high shear mixer and subsequently granulated. The hot granules were allowed to cool to room temperature and sieved through a sieve with a mesh size of 1.0 mm. The granules are blended with partially crosslinked povidone and microcrystalline cellulose in a suitable mixer. Magnesium stearate was added to this blend after the lumps were removed by a 1.0mm sieve. The final blend is then produced by final blending in a suitable mixer and compressed into lozenges. The following lozenge compositions were obtained:

3. Lozenge formulation-Hot melt granulation

The active CCR3 antagonist ingredients, mannitol (mannit), polyethylene glycol, and partial crospovidone were blended in a suitable mixer to create a pre-mix. The premix is heated in a high shear mixer and subsequently granulated. The hot granules were allowed to cool to room temperature and sieved through a sieve with a mesh size of 1.0 mm. The granules were blended with partially cross-linked povidone and mannitol in a suitable mixer. Magnesium stearate was added to this blend after the lumps were removed by a 1.0mm sieve. The final blend is then produced by final blending in a suitable mixer and compressed into lozenges. The following lozenge compositions were obtained:

4. lozenge formulation-Hot melt extrusion

The active CCR3 antagonist ingredients and stearic-palmitic acid were blended in a suitable mixer to create a pre-mix. The premix is extruded in a twin-screw extruder and subsequently granulated. The granules were sieved through a sieve with a mesh size of 1.0 mm. The granules were mixed with mannitol and crospovidone in a suitable mixer. Magnesium stearate was added to this blend after the lumps were removed by a 1.0mm sieve. The final blend is then produced by final blending in a suitable mixer and compressed into lozenges. The following lozenge compositions were obtained:

5. Lozenge formulation-Hot melt extrusion

The active CCR3 antagonist ingredients and stearic-palmitic acid were blended in a suitable mixer to create a pre-mix. The premix is extruded in a twin-screw extruder and subsequently granulated. The granules were sieved through a sieve with a mesh size of 1.0 mm. The granules were directly filled into hard capsules. The following capsule compositions were obtained:

components	mg/lozenge	A tablet
			Active ingredient	70.000	70.0
Stearic acid-palmitic acid	30.000	30.0
			In total (granules)	100.000	100.0
Capsule	90.000	-
			Total of	190.000	100.000

6. Lozenge formulation-roller compaction

The active CCR3 antagonist ingredients, part of the mannitol, and crospovidone and magnesium stearate were blended in a suitable mixer to create a pre-blend. The premix was compacted using a roller compactor and subsequently granulated. Optionally, the granules are sieved through a sieve with a mesh size of 0.8 mm. The granules were blended with a portion of mannitol and crospovidone in a suitable mixer. Magnesium stearate was added to this blend after the lumps were removed by a 1.0mm sieve. The final blend is then produced by final blending in a suitable mixer and compressed into lozenges. The following lozenge compositions were obtained:

components	mg/lozenge	A tablet
			Active ingredient	28.500	30.0
Cross-linked polyvidone	1.400	1.5
			Mannitol	34.600	36.4
Magnesium stearate	0.500	0.5
			In total (granules)	65.000	68.4
Mannitol	27.000	28.4
			Co-polyvidone	1.600	1.7
Cross-linked polyvidone	0.950	1.0
			Magnesium stearate	0.450	0.5
Total of	95.000	100.000

7. Lozenge formulation-roller compaction

The active CCR3 antagonist ingredient and magnesium stearate are blended in a suitable mixer to create a pre-mix. The premix was compacted using a roller compactor and subsequently granulated. Optionally, the granules are sieved through a sieve with a mesh size of 0.8 mm. The granules were blended with mannitol and croscarmellose sodium in a suitable mixer. Magnesium stearate was added to this blend after the lumps were removed by a 1.0mm sieve. The final blend is then produced by final blending in a suitable mixer and compressed into lozenges. The following lozenge compositions were obtained:

components	mg/lozenge	A tablet
			Active ingredient	114.200	66.0
Magnesium stearate	1.800	1.0
			In total (granules)	116.000	67.0
Mannitol	51.000	29.5
			Croscarmellose sodium	3.500	2.0
Magnesium stearate	2.500	1.5
			Total of	173.000	100.000

8. Lozenge formulation-roller compaction

The active CCR3 antagonist ingredient and magnesium stearate are blended in a suitable mixer to create a pre-mix. The premix was compacted using a roller compactor and subsequently granulated. Optionally, the granules are sieved through a sieve with a mesh size of 0.8 mm. The granules are blended with microcrystalline cellulose and crospovidone in a suitable mixer. Magnesium stearate was added to this blend after the lumps were removed by a 1.0mm sieve. The final blend is then produced by final blending in a suitable mixer and compressed into lozenges. The following lozenge compositions were obtained:

9. Coated lozenge formulation

Dragee cores according to the above formulation can be used for the production of film-coated dragees. Hydroxypropyl methylcellulose, polyethylene glycol, talc, titanium dioxide and iron oxide are suspended in purified water in a suitable mixer at ambient temperature to produce a coating suspension. The tablet cores were coated with the coating suspension to a weight gain of about 3% to produce film coated tablets. The following film coating composition can be obtained:

b. pharmaceutical formulation and administration

The study product of the invention (compound 1) conforms to the following chemical structure:

compound 1.

One of ordinary skill in the relevant art will recognize that the compounds, co-crystals, salts, and formulations described in the above sections are also useful in these examples.

Compound 1 can be made into 100mg, 200mg and 400mg film-coated tablets having a biconvex, rounded or oval shape and a dull red color. These tablets are produced by a dry granulation process and contain, as inactive ingredients, microcrystalline cellulose, hydrogen phosphate, croscarmellose sodium, magnesium stearate, polyvinyl alcohol, titanium dioxide, polyethylene glycol, talc, red iron oxide and yellow iron oxide. Placebo lozenges matching the study products were produced by direct compression method and contained the same inactive ingredients.

c. Preclinical examples

1. Materials and methods

(a) Subcutaneous osmotic pump implantation

The day before implantation, the Alzet micropump was filled, prepared and numbered as mouse ID to allow priming at 37 ℃ and to allow blind treatment. The pump was implanted in the back, slightly posterior to the scapula and slightly off-center. Mice were anesthetized with 3 to 5% isoflurane using a vaporizer and regulator in an induction room, then moved to the surgical area and nose cone mounted to maintain 1 to 3% isoflurane anesthesia. The eye ointment is applied to the eye to prevent dryness. Mice were injected subcutaneously with meloxicam (meloxicam)5 mg/kg. The hair was removed from the incision area using a small sharp scissors and the area was cleaned using alternate applications of 70% isopropyl alcohol and betadine (betadine). A 0.5 to 1cm incision is made and a hemostat is inserted to deploy the subcutaneous tissue to form a pump pocket. A pump is inserted into the pocket and the wound is closed with a wound clip. All surgical tools were autoclaved on the day of surgery before first use. Subsequently, the instruments were sterilized between animals using a glass bead sterilizer. Mice were placed in a clean recovery cage partially placed on top of a heating pad until fully recovered and ambulatory. Mice were tested for induction of anesthesia by toe-pinch and monitoring respiration. Mice were monitored every 15 minutes post-surgery until recovery. The following day the mice were administered a second dose of meloxicam. If signs of infection are observed, mice are given subcutaneous 5mg/kg Rakatril (Baytril) daily until the infection is cleared.

(b) Open space Test (Open Field Test)

Open space is used to evaluate general autonomic activity and exploratory behavior in new environments. At least 30 minutes prior to testing, mice were brought to the laboratory to adapt to laboratory conditions (dim lighting). The test site consisted of a 50cm x 50cm square site. Mice were placed in the center of the field and followed for 15 minutes. The time spent in the periphery and the central area and the development behavior were analyzed. All surfaces between the tests were cleaned using 70% ethanol.

(c) Y-shaped labyrinth

The large Y-maze test assesses short-term memory of familiarity with a particular context. At least 30 minutes prior to testing, mice were brought to the laboratory to adapt to laboratory conditions (dim lighting). For the initial training trial, mice were placed on the end of one arm of a large Y-shaped maze designated the "starting arm" (arm length: 15 inches). The third arm of the maze was blocked, allowing the mouse to freely explore two of the three arms ("the initial arm" and the "familiar arm") for 5 minutes. Each arm contains a spatial cue. One hour later, the mice were placed back in the "starting arm" of the maze and allowed to explore all three arms, with the third arm not blocked ("novel arm"). The movement into and out of each arm is tracked using automated tracking software (CleverSys). The tests were performed under dim lighting conditions and the equipment was cleaned with 70% ethanol between trials.

(d) Barnes Maze (Barnes Maze)

The modified bahness maze is used to evaluate spatial work/scenarios like learning and memory. The Barnes maze was composed of a 122cm diameter circular platform with 40 escape holes, each hole 5cm in diameter, placed along three rings at different distances from the center of the platform. One of these holes is accompanied by an escape box, all of which are uncovered. Training with lights and fans in the maze provides adverse stimuli to assist in escape. Visual cues were placed on all four sides of the maze. Mice were provided with a series of 4 or 5 trials with trial intervals of approximately 10 minutes and a maximum duration of 90 or 120 seconds for each trial. For each experiment, the mice were placed in the center of the maze. After 10 seconds, the mice were allowed to explore and the experiment was ended if they found the escape box and entered it before the end of the experiment. The mice that could not find the escape box were guided to the escape box and allowed to enter, and stayed for 30 seconds before being returned to their home cages. Training lasted 4 days. The data recorded and analyzed include speed, escape latency and travel distance.

The mice were divided into groups of 4 to 5 mice each, and the treatment groups were equilibrated. For example, group 1 mice were tested 4 times, followed by group 2 mice 4 times, and so on until all groups were tested. The field was cleaned between trials using 70% ethanol.

(e) Rotating rod method

Mice were trained 3 trials (up to 100 seconds each) in the rotarod method, which is a test of motor coordination. Success or failure was recorded in the last trial, with success defined as drop latency >90 seconds. Two tests were performed to compare the success rate between control and compound-treated mice.

(f) T-shaped labyrinth

The water maze was filled with water at least 24 hours before the test to allow it to reach room temperature. The water was stained with white latex paint to make the animals visible for tracking and to allow the use of a hidden platform. Two different visual cues were placed at the ends of the two T-arms of the T-maze insert. On day 1, animals were provided with 4 trials, each with a visible plateau and a 30 minute inter-trial interval. Animals were offered 60 seconds to reach the platform. If they do not reach the platform within that time, they are directed to the platform and allowed to remain for 5 seconds and then removed from the water tank. Target arms were switched after every three mice and both treatment groups had the same number of right and left turning target arms. After each trial, mice were placed in empty cages with blue pads and allowed to dry under red light before being returned to their home cages. Day 2 is the test day, where animals were subjected to the same test, 4 trials each and 30 minute inter-trial intervals, but with a hidden platform. Animals were scored for correct or incorrect selection and latency to reach the platform and two tests were performed to compare the success rate between control and compound-treated mice. All experiments were recorded using TopScan.

(g) Protein quantification

CCL11 protein levels were measured in mouse plasma by sandwich ELISA. Plasma was diluted 1:10 for the assay. (mouse CCL11/Eotaxin Duo Set ELISA kit, R & D Systems, Minneapolis, MN).

Human CCL11 was measured in human plasma by a SomaLogic aptamer-based assay (SOMAscan, available from SomaLogic, inc., Boulder, CO). (Gold L et al, (2010) Aptamer-Based Multiplexed technical for Biomarker discovery. PLOS ONE 5(12): e 15004).

Mouse plasma was analyzed by Luminex for a panel of circulating cytokines. Luminex Assay Service was performed by Eve Technologies (Calgary, Alberta, Canada).

(h) mRNA quantitation

Dissecting the frozen brain into cortex, hippocampus, striatum and thalamus. The cortex was divided into three homogeneous fractions, and one fraction was used for RNA isolation. RNA was isolated using the Prolink RNA Mini Kit (Thermo Fisher Scientific # 12183025). cDNA was prepared using the Taqman RT Kit (Thermo Fisher Sci # N8080234). qPCR was run on Quant Studio 6 using custom Taqman Multiplex primers for IL-1 β and GAPDH. All samples were run together on a single plate and analyzed first for ddCT against their endogenous controls and then against the control group.

(i) HPLC/MS quantification of Compound 1

Compound levels were measured by MS/MS of Quintara (Hayward, Calif.).

(j) Flow analysis

Eosinophil count

Charles River Clinical Pathology Services (Shrewsbury, Mass.) collecting 200. mu.L of whole blood during perfusion and shipping were used for hematology analysis of eosinophil counts by FACS (fluorescence activated cell sorter).

Eosinophil Shape Change (ESC)

ESCs determined the shape change of human eosinophils activated by human eotaxin-1 (PreProTech, Rocky Hill, New Jersey) compared to native eosinophils. This change was detected as a change in forward scatter as measured by FACS (fluorescence activated cell sorter). The mean forward scatter of the autofluorescent (eosinophil) population of each sample was determined in combination with the mean of triplicates for each set of samples. Methods of determining ESCs have been described previously and are known in the art.

CCR3 receptor internalization

CCR3 receptor internalization assay is FACS-based and uses human eosinophilsGranulocyte chemokine-1 (Eotaxin-1) (PreProTech, Rocky Hill, New Jersey) and by APC (R&D Systems, Minneapolis, Minnesota) labeled anti-human CCR3 antibody or isotype control IgG ₂Antibody a was used to monitor CCR3 receptor internalization in human eotaxin-1 induced human eosinophils as compared to untreated eosinophils. Median APC fluorescence intensity units for each sample and the average of triplicates for each set of samples were determined. Methods of monitoring internalization of the CCR3 receptor have been described previously and are known in the art.

(k) Histology

The next day after the behavioral testing was completed, the mice were removed. Anesthesia was induced by 2,2, 2-tribromoethanol and then perfused with 0.9% saline via the heart. The brain was dissected and cut in two uniform halves in the sagittal plane. Half were snap frozen in dry ice for later use, and the other half was fixed in 4% paraformaldehyde in PBS for immunohistochemistry. Two days after fixation, the hemibrain was transferred to a PBS solution containing 30% sucrose and replaced two days later. The brains were sectioned at 30um on a microtome at-22 ℃. Brain sections were stored in cryoprotectant media at-20 ℃ until staining. Blocking was performed on floating sections in 10% serum contained in 0.5% appropriate serum from PBST. The primary antibody was incubated overnight at 4 ℃. For optical microscopy, the following antibodies were used at given concentrations: DCX, 1:200, Santa Cruz BioTech, CD68, 1:1000, AbD Serotec. The following day secondary biotinylated antibody was administered at a concentration of 1: 300. Visualization of staining was achieved by reaction with ABC kit (Vector) and diaminobenzidine (Sigma). Dehydration of the mounted slides was achieved using ethanol and xylene dipping. Images were taken at 5 x magnification on a Leica optical microscope.

For fluorescence microscopy, the following antibodies were used at given concentrations: GFAP, 1:500, DAKO; iba1, 1:1000, Wako; and BrdU, 1:500, AbCam. An antigen repair protocol for BrdU (2N HCl, 37C, 30 min) was required prior to blocking. The following day the appropriate fluorescent secondary antibody was administered at a concentration of 1:300 for 1 hour at room temperature. The slides were covered with an elongated gold mounting medium. Images were taken on an optical microscope at 5 x magnification.

2. Experimental group

First experimental group (see fig. 1 to 2): 2-or 18-month-old C57Bl/6 mice were administered either by IP injection via IgG antibody control or subcutaneously via compound 1 via Alzet osmotic pump for 2 or 4 weeks. During the last week of treatment, mice were subjected to behavioral testing prior to perfusion on the last day of treatment. Immediately prior to the start of treatment, all mice received 150mg/kg BrdU IP injections for 5 consecutive days.

And (5) blending the medicines. Control rat IgG₂Pure A54447 (MAB006, R)&D Systems) was administered 50ug/kg in sterile saline. Compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Fresh solutions were prepared weekly and stored at 4 ℃.

Treatment group:

treatment group 1, young control: young C57BL/6 mice (n ═ 18) aged 1 to 2 months received 5 injections of control IgG by Intraperitoneal (IP) injection, once every 3 days for 14 days.

Treatment group 2, geriatric control: older C57BL/6 mice (n ═ 18) aged 18 months received 5 injections of control IgG by IP injection, every 3 days for 14 days.

Treatment group 3, compound 1 (dose 1) aged: older C57BL/6 mice (n ═ 16) at the age of 18 months received-50 mg/ml infusions of compound 1 by Alzet minipump type 2001 (1uL/hr) for two weeks, and were replaced once.

Treatment group 4, compound 1 (dose 2) old: older C57BL/6 mice (n ═ 16) at the age of 18 months received-50 mg/ml infusions of compound 1 by Alzet mini pump model 2002 (0.5uL/hr) for two weeks.

Treatment group 5, compound 1 (dose 2) old: older C57BL/6 mice (n ═ 16) at the age of 18 months received-50 mg/ml infusions of compound 1 by Alzet mini pump model 2002 (0.5uL/hr) for four weeks, and were replaced once.

Subcutaneous administration of compound 1 at four weeks in 18-month old mice increased the number of double-cortin positive cells in the hippocampus, which are indicative of neurogenesis (see figure 1A). Administration of higher doses of compound 1 for 2 weeks resulted in an increased propensity for BrdU positive cells in the hippocampus, which are markers of cell proliferation (see fig. 1B). Compound 1 low or high infusion for 2 or 4 weeks resulted in an improvement in the suggestive Y-maze (a memory test) (see figure 2, reports normalized to percentage of time of total interaction time and number of visits-similar effects are also seen in total time spent evaluating). Comparison was made with control groups (treatment groups 1 and 2) treated with IgG antibody without compound and administered and behaviours were performed in parallel.

Thus, administration of compound 1 increased the number of Dcx and BrdU positive cells, indicating that compound 1 increased neurogenesis and cell survival, respectively. Compound 1 was able to improve memory (cognition) as demonstrated by performance in the Y-maze test.

Second experimental group (see fig. 3 to 6): c57Bl/6 mice 3 or 16.5 months of age were administered subcutaneously via vehicle control or compound 1 for 4 weeks by an Alzet osmotic pump. During the last week of treatment, mice were subjected to behavioral testing prior to perfusion on the last day of treatment. Immediately prior to the start of treatment, all mice received 150mg/kg BrdU IP injections for 5 consecutive days.

And (5) blending the medicines. Compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The carrier solution was similarly prepared and the pH was adjusted. Fresh solutions were prepared weekly and stored at 4 ℃.

Treatment group:

treatment group 1, young control: young C57BL/6 mice (n ═ 19) aged 3 months received vector infusions by Alzet mini pump model 2002 (0.5uL/hr) for four weeks, and were replaced once.

Treatment group 2, geriatric control: older 16-month-old C57BL/6 mice (n ═ 19) received vector infusion by Alzet mini pump model 2002 (0.5uL/hr) for four weeks, and were replaced once.

Treatment group 3, compound 1 (dose 2) old: aged 16-month old C57BL/6 mice (n ═ 19) received an infusion of-50 mg/ml compound 1 by Alzet mini pump model 2002 (0.5uL/hr) for four weeks, and were replaced once.

Subcutaneous administration of compound 1 improved four weeks in 16.5 month old mice suggesting cognitive performance for memory in the Y-maze test (see figure 3). Four-week administration of compound 1 also tended to improve performance in the Barnesian maze, a hippocampus-dependent spatial memory test (see FIG. 4). A tendency to improve memory was also observed by analyzing the mean latency of the day 4 trial and the difference between the latencies of trials 13 and 16. Four weeks of administration of compound 1 also significantly increased the number of double BrdU positive cells in the hippocampus, which are indicative of neurogenesis (see figure 5). Thus, compound 1 was able to simultaneously improve cell survival and improve memory (cognition) as demonstrated by the results obtained using the Y-maze and the barnes maze tests.

Compound cerebrospinal fluid levels in mice

Cerebrospinal fluid (CSF) was collected from the 2-month old "young" and 16.5-month old "groups and the level of compound 1 was determined by mass spectrometry. Figure 6 depicts the levels of compounds of the invention detected in mouse CSF for the young and old groups (both below 10 nM). These CSF levels do not approximate the Ki of the compound in mice (124nM, determined by cell line receptor binding) and therefore do not cross the Blood Brain Barrier (BBB) at significant concentrations. For comparison, the levels of the compounds of the invention measured in young (2 month old) and old (18 month old mice) plasma perfused with 0.5 μ L/hr of 50mg/mL solution for 2 and 4 weeks, respectively, were significantly higher (352. + -.31 nM and 355. + -.43 nM, respectively; values are mean. + -..s.e.m.), which further indicates that the compounds of the invention were not able to cross the BBB in significant amounts.

This data shows that compound 1 does not act directly on the CNS, and therefore acts peripherally. In addition, the concentration across the BBB is not sufficient to make it effective. Furthermore, BBB penetration was not different between young and old mice, indicating that compound 1 exhibited effects on cognitive and neurodegenerative diseases not attributable to differences in BBB between the two groups.

Level of tissue distribution of compound

In oral administration of the warp 2¹⁴C]After radiolabeling compound 1 ("labelled compound") to male-stained C57BL/6JOlaHsd mice (Harlan Labs, BV),the distribution of compound 1 in mouse tissues was determined. The labeled compound was administered at 10mg/kg body weight, corresponding to 17. mu. mol/kg. One animal was killed 1, 24 and 168 hours after application. Blood, plasma and ocular radioactivity concentrations were measured using Liquid Scintillation Counting (LSC). Tissue and organ concentrations were measured by whole body automated radiography (QWBA). According to known techniques (see S.Ullberg et al, Autoadiography in Pharmacology, The int.encyclopedia of Pharmacology, J.Cohen (eds.), 1(78):221-39(1971)), using an extremely cold temperature microtome Reichert-Jung CRYO MACROUT or CRYO MACROUT LEICA CM 3600³Whole body animal sections were prepared.

The following sections were taken at different levels by embedding animals and whole body sections at 5 to 7 levels were selected to allow quantitative assessment of radioactivity: the adrenal gland; blood; bone marrow; a brain; the eye (lens); epididymis; fat (white and brown); harderian glands (Harderian gland); a heart; the kidney; liver; a lung; a muscle; a pituitary; the pancreas; prostate, spinal cord; a spleen; salivary glands; skin; a testis; thyroid gland; thymus; grape membrane. Two sections per animal were taken for each selected level and those sections were lyophilized in a microtome at-20 to-25 ℃ for a minimum of 48 hours.

Fig. 7 depicts a graph reporting values of area under the curve (AUC) for all three time points and quantifies exposure of each tissue to the marker POI after compound administration. Figure 7 shows that compound 1 does not cross the Blood Brain Barrier (BBB) at significant levels.

Again, these results show that compound 1 acts in a peripheral manner because it cannot cross the BBB at appreciable levels. Thus, the effect of compound 1 does not directly act on the central nervous system, and it overcomes the difficulties leading to the failure of many drug candidates targeting CNS diseases.

Pharmacokinetic profile of oral administration

At two doses: concentrations of Compound 1 in the plasma of male 2-month old C57Bl/6 mice were measured at time points 20 minutes, 2 hours, 8 hours, and 12 hours after oral gavage at 30mg/kg and 150 mg/kg. A dose of 30mg/kg was found to be sufficient to achieve a concentration of compound 1 of greater than 100nM at 8 hours (figure 8).

Third experimental group (see fig. 9 to 11): 2-month-old C57Bl/6 mice were administered twice daily oral gavage for 18 days with either vehicle control or Compound 1. During the last week of treatment, mice were subjected to behavioral testing prior to perfusion on the last day of treatment. Immediately prior to the start of treatment, all mice received 150mg/kg BrdU IP injections for 5 consecutive days.

Preparing the medicines: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The carrier solution was similarly prepared and the pH was adjusted. Fresh solutions were prepared weekly and stored at 4 ℃.

Treatment group:

treatment group 1a, support treatment: young C57BL/6 mice (n ═ 15) 7 weeks old received oral (PO) treatment of the vehicle solution twice daily (BID) for 18 days, with only 1 injection on the last day for a total of 35 injections.

Treatment group 1b, compound 1 treatment: young C57BL/6 mice (n ═ 15) 7 weeks of age received oral (PO) treatment of compound 1 twice daily (BID) for 18 days, with only 1 injection on the last day for a total of 35 injections.

Treatment group 2a, vector with rmCCL 11: young C57BL/6 mice (n ═ 15) 7 weeks old received oral (PO) treatment of the vehicle solution twice daily (BID) for 18 days, with only 1 injection on the last day for a total of 35 injections. Mice received peripheral Injections (IP) of rmCCL11 every 3 days for 5 injections simultaneously starting on day 1 of treatment.

Treatment group 2b, compound 1 with rmCCL 11: young C57BL/6 mice (n ═ 15) 7 weeks old received oral (PO) treatment of compound 1 twice daily for 18 days, with only 1 injection on the last day for a total of 35 injections. Mice received peripheral Injections (IP) of rmCCL11 every 3 days for 5 injections simultaneously starting on day 1 of treatment.

Recombinant mouse CCL11 ("rmE") treatment significantly aggravated anxiety in the open space, but improved anxiety with compound 1 orally for 2 weeks twice daily (see figure 9). rmCCL11 impaired memory in the Y-maze; however, the mice treated with compound 1 were no longer significantly different from the control mice (see figure 10). rmCCL11 also impaired memory in the bahnes maze, and treatment with compound 1 significantly improved memory performance (see figure 11). Thus, rmE aggravated both anxiety and memory via the open space test as evidenced by the performance measured in the Y-maze and the banus-maze tests. However, compound 1 treatment can attenuate these effects.

Fourth experimental group (see fig. 12): 23-month-old C57B1/6 mice were administered twice daily oral gavage for 19 days with either vehicle control or Compound 1. After 11 days of treatment, mice were subjected to the Y-maze test.

Preparing the medicines: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The carrier solution was similarly prepared and the pH was adjusted. Coriver (Kolliphor) was added to each solution at 10% per week and stored at 4 ℃.

Treatment group:

treatment group 1, support treatment: older 23-month-old C57B1/6 mice (n ═ 8) received twice daily (BID) treatment with oral gavage (PO) of the carrier solution for 19 days.

Treatment group 2, compound 1 treatment: older 23-month-old C57B1/6 mice (n ═ 11) received oral gavage (PO) treatment of compound 1 twice daily (BID) at 30mg/kg for 19 days.

Compound 1 treatment significantly improved memory in the Y-maze. Compound 1-treated mice exhibited intact memory for the novel arm in the number of visits (fig. 12A). Treatment with compound 1 also significantly increased the travel distance of mice during the test (fig. 12B).

Fifth experimental group (see fig. 13 to 17): 23-month-old C57B1/6 mice were subcutaneously administered twice daily with vehicle control or Compound 1 for 21 days. Three weeks later, mice were subjected to behavioral testing and sacrificed the next day of the last behavioral testing.

Preparing the medicines: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The carrier solution was similarly prepared and the pH was adjusted. Coriver (Kolliphor) was added to each solution at 10% per week and stored at 4 ℃.

Treatment group:

treatment group 1b, support treatment: older 23-month-old C57B1/6 mice (n ═ 9) received Subcutaneous (SQ) treatment with vehicle solution twice daily (BID) for 21 days.

Treatment group 4, compound 1 treatment: older 23-month-old C57B1/6 mice (n ═ 17) received Subcutaneous (SQ) treatment of compound 1 twice daily (BID), 30mg/kg, for 21 days.

Compound 1 treatment significantly improved memory in the Y-maze and the barnes maze. Compound 1-treated mice exhibited complete memory for the novel arm on the number of visits (fig. 13A-B) and the duration of time spent in the novel arm (fig. 13C-D). Treatment with compound 1 also significantly increased the speed of mice during the test (fig. 13E). Compound 1-treated mice performed significantly better in the bahnes maze on spatial memory (fig. 14A), and also showed increased speed during the test (fig. 14B). Autonomic activity in open space is also improved, with a strong tendency to improve travel distance and speed (fig. 15A and 15B, respectively).

Inflammatory cytokine levels in mouse plasma were measured by Luminex assay (figure 16). There is a strong tendency for several inflammatory markers to decline, including TNF α, IL6, IL1 β, IL5 and IL 17. Activated microglia were also quantified by IHC staining in the hippocampus of mice (fig. 17). Compound 1 treatment resulted in a strong tendency for microglial depletion.

Sixth experimental group (see fig. 18): 23-month-old C57B1/6 mice were subcutaneously administered twice daily for 30 days with vehicle control or Compound 1 and subsequently sacrificed on the day of the last injection.

Preparing the medicines: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The carrier solution was similarly prepared and the pH was adjusted. Coriver (Kolliphor) was added to each solution at 10% per week and stored at 4 ℃.

Treatment group:

treatment group 1a, support treatment: older 23-month-old C57B1/6 mice (n ═ 9) received Subcutaneous (SQ) treatment of the vehicle solution twice daily (BID) for 30 days.

Treatment group 3, compound 1 treatment: older 23-month-old C57B1/6 mice (n ═ 18) received Subcutaneous (SQ) treatment of compound 1 twice daily (BID) at 30mg/kg for 30 days.

FACS analysis in subgroups of animals (n-2, 5) revealed that compound 1 treatment resulted in a strong tendency for blood eosinophilia depletion in blood (fig. 18).

Seventh experimental group (see fig. 19): every other day for 3-month-old hairless miceOxazolone (Ox) treatment and treatment with control saline or 30mg/kg compound 1BID, PO for 2 weeks.

Preparing the medicines: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The carrier solution was similarly prepared and the pH was adjusted. Colift was added to each solution at 10% per week and stored at 4 ℃.

Treatment group:

treatment group 1, support treatment: 3-month-old hairless mice (n ═ 3) received oral gavage (PO) treatment of the carrier solution twice daily (BID) for 2 weeks.

Treatment group 2, vector treatment andand (3) azolone treatment: 3-month-old hairless mice (n ═ 6) received oral gavage (PO) treatment of the carrier solution twice daily (BID) for 2 weeks.

Treatment group 3, Compound 1 treatment andand (3) azolone treatment: 3-month-old hairless mice (n ═ 6) received oral gavage (PO) treatment of compound 1 twice daily (BID), 30mg/kg, for 2 weeks.

Compound 1 was shown to decrease in total blood count (CBC) analysisOxazolone-induced blood eosinophils Is increased. N is 3, 6, 8. P<0.05，**p<0.01. In thatCompound 1 significantly reduced the number of blood eosinophils in the oxazolone-induced eosinophilia model (fig. 19). This shows that inhibition of CCR3 may be sufficient to normalize eosinophil levels and function, especially in diseased states, demonstrating that compound 1 may be effective in treating a secondary mechanism of neuronal loss and associated motor and cognitive deficits (in addition to reducing brain inflammation).

Eighth experimental group (see fig. 20 to 22): 24-month old C57 mice were treated for 4 weeks with continuous infusion of compound 1 or vehicle by Alzet osmotic pumps implanted to deliver both treatments continuously.

Preparing the medicines: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The carrier solution was similarly prepared and the pH was adjusted. Colift was added to each solution at 10% per week and stored at 4 ℃.

Treatment group:

treatment group 1, geriatric control: older 23-month-old C57BL/6 mice (n ═ 15) received vector infusion by Alzet mini-pump model 2002 (0.5uL/hr) for four weeks, and were replaced once.

Treatment group 2, compound 1 (dose 2) old: older 23-month-old C57BL/6 mice (n ═ 15) received an infusion of-50 mg/ml compound 1 by Alzet mini-pump model 2002 (0.5uL/hr) for four weeks, and were replaced once.

Mice were tested for motor coordination in the rotarod method. Success or failure was recorded in the last trial, with success defined as drop latency >90 seconds. By both tests, mice treated with compound 1 were more successful than vehicle treated mice. Each N15, p < 0.05. A higher proportion of compound 1-treated mice (47%) were able to stay on the rods for more than 90 seconds, with only 20% of the control-treated mice being able to maintain this threshold after 3 consecutive trials (fig. 20). These results indicate that compound 1 has a consistent effect on motor function in the eotaxin-elevated model.

Mice were tested for cognitive function in a T-maze (figure 21). The number of successes or failures to lower the correct arm is recorded. By both tests, mice treated with compound 1 were more successful than vehicle treated mice. P < 0.05.

Fecal output was measured by weighing the dry weight of overnight fecal pellets (fig. 22). Gastric function is a well-known symptom of parkinson's disease. Water and food intake were measured over the same time period. Compound 1-treated mice had significantly lower fecal output compared to control mice. Their overnight water intake was also significantly higher compared to control mice. P < 0.05. These results indicate that compound 1 treatment may alter defects in gastric function.

Ninth experimental group (see fig. 23): three-month-old C57 mice were given a single dose of 10mg/kg IP Lipopolysaccharide (LPS) to induce inflammation and treated with Compound 1 for 18 days.

Preparing the medicines: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The carrier solution was similarly prepared and the pH was adjusted. Colift was added to each solution at 10% per week and stored at 4 ℃.

Treatment group:

treatment group 1, support treatment: young C57BL/6 mice (n ═ 15) aged 3 months received a single injection of LPS and were treated with vehicle by oral gavage, BID, for 18 days.

Treatment group 2, compound 1 treatment: young C57BL/6 mice (n ═ 15) aged 3 months received a single injection of LPS and were treated with compound 1 by oral gavage, 30mg/kg, BID, for 18 days.

Brain sections were immunostained for detection of CD68+ activated microglia. Compound 1-treated mice showed significantly reduced CD68+ immunoreactivity (decreased activated microglia) compared to vehicle (saline) -treated mice (fig. 23). N is 9, 10, 8. P <0.05 by one-factor ANOVA.

These data indicate a potent anti-neuroinflammatory effect of compound 1 with therapeutic potential to reduce neuroinflammation-induced neuronal toxicity in diseases that show neurodegeneration or cognitive or motor decline.

Tenth experimental group (see fig. 24 to 29): three-month-old C57 mice were provided with a daily dose of 0.5mg/kg IP Lipopolysaccharide (LPS) to induce inflammation and were chronically treated with compound 1 for up to 4 weeks.

Preparing the medicines: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The carrier solution was similarly prepared and the pH was adjusted. Colift was added to each solution at 10% per week and stored at 4 ℃.

Treatment group:

treatment group 1, support treatment: 3-month-old C57 mice (n ═ 10) received treatment with LPS daily IP injection for 7 weeks and twice daily (BID) oral gavage (PO) carrier solution for up to 4 weeks.

Treatment group 2, vector treatment andand (3) azolone treatment: 3-month-old C57 mice (n ═ 10) received treatment with LPS daily IP injection for 7 weeks and twice daily (BID) oral gavage (PO) carrier solution for up to 4 weeks.

Treatment group 3, Compound 1 treatment andand (3) azolone treatment: 3-month-old C57 mice (n ═ 9) received treatment with LPS daily IP injection for 7 weeks and twice daily (BID) oral gavage (PO) of compound 1, 30mg/kg, for up to 4 weeks.

Fig. 24A depicts a dosing paradigm for the tenth experimental group. The example portion highlighted by the square indicates the time point when the examination in fig. 24B is performed. Anxiety was tested in the open space assay 1 week after compound 1 treatment (fig. 24B). LPS treatment significantly increased anxiety in the open space, while compound 1 strongly reduced increased anxiety, p < 0.05.

Fig. 25A depicts a dosing paradigm for the tenth experimental group. The example portion highlighted by the square indicates the time point when the examination in fig. 25B is performed. Cognition was tested in the Y-maze 3 weeks after compound 1 treatment (fig. 25B). LPS-treated mice did not show significant preference for the novel arm. However, similar to vehicle-treated mice, compound 1-treated mice showed significant preference for novel arms, # p <0.05, # p < 0.01.

Fig. 26A depicts a dosing paradigm for the tenth experimental group. The example portion highlighted by the square indicates the time point at which the detection timing in fig. 26B is performed. mRNA levels of the inflammatory cytokine IL-1 β were measured by quantitative PCR 4 weeks after compound 1 treatment (fig. 26B). LPS-treated mice showed a tendency to increase IL-1 β levels, whereas compound 1-treated mice showed a significant decrease in IL-1 β expression, p < 0.05.

Fig. 27A depicts a dosing paradigm for the tenth experimental group. The example portion highlighted by the square indicates the time point when the examination in fig. 27B is performed. Brain sections were immunostained for detection of CD68+ activated microglia. Compound 1-treated mice showed significantly reduced CD68+ immunoreactivity (decreased activated microglia) compared to LPS-only-treated mice (fig. 27B).

Fig. 28A depicts a dosing paradigm for the tenth experimental group. The example portion highlighted by the square indicates the time point when the examination in fig. 28B is performed. Brain sections were immunostained for Iba1 positive microglia. Compound 1 treated mice showed significantly reduced Iba1+ immunoreactivity (total microglia reduction) compared to LPS only treated mice (fig. 28B).

Fig. 29A depicts a dosing paradigm for the tenth experimental group. The example portion highlighted by the square indicates the time point when the examination in fig. 29B is performed. Brain sections were immunostained for detection of GFAP positive astrocytes. Mice treated with compound 1 showed significantly reduced GFAP + immunoreactivity (total astrocytes decreased) compared to mice treated with LPS alone (fig. 29B).

These data indicate a potent anti-neuroinflammatory effect of compound 1 with therapeutic potential to reduce neuroinflammation-induced neuronal toxicity in diseases that show neurodegeneration or cognitive or motor decline.

Eleventh experimental group (see fig. 30): forty-five (42)2 month old C57BL/6 mice were divided into three groups: vehicle treated controls, vehicle treated LPS mice and compound 1 treated LPS mice. The vehicle or compound (PO BID) was administered orally twice daily for 6 doses. Hippocampus was evaluated histologically with the microglial marker Iba-1.

Preparing the medicine: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The vehicle solution was prepared and the pH was similarly adjusted. Solutions were prepared fresh weekly and stored at 4 ℃.

And (3) LPS preparation: LPS was formulated at 0.55mg/ml in saline and administered at 5 mg/kg.

Treatment group:

·treatment group 1Vehicle treatment control: young C57BL/6 mice 2 months old (n ═ 13) received a single IP saline injection followed by vehicle PO BID for 3 days for 6 injections.

·Treatment group 2Vehicle-treated LPS: young C57BL/6 mice 2 months old (n ═ 19) received a single IP injection of LPS (5mg/kg) followed by vehicle PO BID for 3 days for 6 injections.

·Treatment group 3Compound 1 treatment of LPS: young C57BL/6 mice 2 months old (n ═ 18) received a single IP injection of LPS (5mg/kg) followed by compound 1(30mg/kg) PO BID for 3 days for a total of 6 injections.

Tissue treatment and histology: the mouse half-brain was cut into 30 μm pieces at-22 ℃ on a microtome. Brain slices were collected sequentially into 12 tubes to represent 12 slices of the hippocampus in a given tube. Brain sections were stored in cryoprotective media at-20 ℃ until staining was required. Free floating sections were blocked in the appropriate serum of 10% serum in PBST 0.5%. The primary antibody, Iba1(Wako, 1: 1000), was incubated overnight at 4 ℃. The following day, appropriate fluorescent secondary antibodies were raised at a rate of 1: 300 was applied at room temperature for one hour. The slides were covered using the Prolong Gold Mounting Media.

Quantification of neuroinflammation: iba-1 positive area was quantified as percentage of ROI around the entire hippocampus using a threshold on Image Pro Premier v9.2 software. About 5 sections from each mouse were taken as the average Iba-1 positive area percentage. Statistical significance was tested using the general one-way anova and Dunnett's multiple post-hoc comparison test between treatment groups.

Figure 30 shows the results of histological analysis of mouse brain in an acute model of LPS-induced inflammation. The results show a significant reduction of LPS-induced microglial proliferation in hippocampus of mice treated with compound 1 when treatment with compound 1 and LPS was initiated on the same day. Microglial proliferation was measured by determining the percentage of Iba-1 positive area in hippocampus. Statistical significance was tested using the general one-way anova and Dunnett's multiple post-hoc comparison test between treatment groups (.;. P05;. P001).

Twelfth experimental group (see fig. 31): fifteen (15)2 month old C57BL/6 mice were divided into three groups: vehicle treated controls, vehicle treated LPS mice and compound 1 treated LPS mice. The vehicle or compound (PO BID) was administered orally twice daily for 6 doses. Hippocampus was evaluated histologically with the microglial marker Iba-1.

Preparing the medicine: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The vehicle solution was prepared and the pH was similarly adjusted. Solutions were prepared fresh weekly and stored at 4 ℃.

And (3) LPS preparation: LPS was formulated at 0.55mg/ml in saline and administered at 5 mg/kg.

Treatment group:

·treatment group 1Vehicle treatment control: young C57BL/6 mice 2 months old (n ═ 13) received a single IP saline injection, 72 hours later with vehicle PO BID for 3 days for 6 injections.

·Treatment group 2Vehicle-treated LPS: young C57BL/6 mice 2 months old (n ═ 19) received a single IP injection of LPS (5mg/kg) and 72 hours later vehicle PO BID was administered for 3 days for a total of 6 injections.

·Treatment group 3Compound 1 treatment of LPS: young C57BL/6 mice 2 months old (n ═ 18) received a single IP injection of LPS (5mg/kg) and compound 1(30mg/kg) PO BID was administered 72 hours later for 3 days for a total of 6 injections.

Tissue treatment and histology: the mouse half-brain was cut into 30 μm pieces at-22 ℃ on a microtome. Brain slices were collected into 12 tubes sequentially to represent each 12 slices of the hippocampus in a given tube. Brain sections were stored in cryoprotective media at-20 ℃ until staining was required. Free floating sections were blocked in the appropriate serum of 10% serum in PBST 0.5%. The primary antibody, Iba1(Wako, 1: 1000), was incubated overnight at 4 ℃. The following day, appropriate fluorescent secondary antibodies were raised at a rate of 1: 300 was applied at room temperature for one hour. The slides were covered using the Prolong Gold Mounting Media.

Quantification of neuroinflammation: iba-1 positive area was quantified as percentage of ROI around the entire hippocampus using a threshold on Image Pro Premier v9.2 software. About 5 sections from each mouse were taken as the average Iba-1 positive area percentage. Statistical significance was tested using the general one-way anova and Dunnett's multiple post-hoc comparison test between treatment groups.

Figure 31 shows the results of histological analysis of mouse brain in an acute model of LPS-induced inflammation. When administration of compound 1 was started three days after administration of LPS, a significant reduction in LPS-induced microglial proliferation in hippocampus of mice treated with compound 1 for three days was shown. Microglial proliferation was measured by determining the percentage of Iba-1 positive area in hippocampus. Statistical significance was tested using the general one-way anova and Dunnett's multiple post-hoc comparison test between treatment groups (.;. P05;. P001).

3. Mouse MPTP model for parkinson's disease

A mouse model using MPTP (1-methyl-4-phenyl-1, 2,3, 6-tetrahydropyridine) was used to determine the level of improvement in MPTP-induced Parkinson's disease deficiency. MPTP is a prodrug of MPP +, which can lead to permanent parkinson's symptoms. MPP + works by killing dopaminergic neurons in the substantia nigra region of the brain.

Seventy-four (74) 8-week-old male C57Bl/6J mice were used together in the experiment. Animals were divided into two separate study arms, 12-day and 4-day study arms, respectively, and similarly treated in both arms. Mice in the study arm were divided into three groups, such that one group served as control, receiving MPTP vehicle and compound vehicle, the other two groups received MPTP twice daily on days 1 and 2, and further received compound vehicle or compound 1(30mg/kg) 1 time daily on days 1 to 12, with only one dose on day 12 (fig. 32). On study day 11, ten days after treatment, motor function of 12 day study arm mice was evaluated in a fine motor analysis. Endpoint tissue treatments were performed on study days 12 and 4. Samples were processed for hematology, immunohistochemistry, and HPLC measurements. Striatal levels of Dopamine (DA), 3, 2-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were assessed by HPLC on day 12. The immunoreactivity of Tyrosine Hydroxylase (TH) was assessed from brain sections collected through the substantia nigra.

Since there are a large number (ninety-seven) of individual parameters, a Principal Component Analysis (PCA) was performed. PCA combines all parameter data together and reveals correlations between different parameters, providing an overall picture of fine motor and gait characteristics. FIG. 33A illustrates a visualization of ten principal components or PCs showing how the original parameters are related in the data set. The greater the intensity (blue or red) of each parameter, the greater the role a particular parameter plays in the corresponding PC. The heatmap is formed from PCAs of all available parametric data. Based on group 2 of all PC scores: MPTP + vehicle and group 1: differences between vehicle + vehicle mice, PC scores are expressed in figure 33B as overall gait analysis scores.

Figure 34 shows the gait characteristics of the anterior paw toe gap on study day 11. The MPTP + vehicle treated group showed statistically significant changes in the anterior paw-toe gap compared to the vehicle + vehicle treated group. And the MPTP + compound 1 treated group showed a positive trend compared to the MPTP + vehicle treated group. Data are presented as mean + SEM (group 1: vehicle + vehicle, n 15; group 2: MPTP + vehicle, n 14; group 3: MPTP + compound 1, n 13). Statistical significance: p <0.05, group 2: MPTP + vehicle control group 1: vehicle + vehicle (unpaired t-test).

Figure 35 shows the gait characteristics of the swing speed of the forepaw at study day 11. The MPTP + vehicle treated group showed a statistically significant change in paw swing velocity compared to the vehicle + vehicle treated group. And the MPTP + compound 1 treated group showed a positive trend compared to the MPTP + vehicle treated group. Data are presented as mean + SEM (group 1: vehicle + vehicle, n 15; group 2: MPTP + vehicle, n 14; group 3: MPTP + compound 1(30mg/kg), n 13). Statistical significance: p <0.05, group 2: MPTP + vehicle control group 1: vehicle + vehicle (unpaired t-test).

Figure 36 shows the gait characteristics of the ankle joint range of motion at study day 11. The MPTP + vehicle treated group showed statistically significant changes in ankle range of motion compared to vehicle + vehicle treated group. And the MPTP + compound 1 treated group showed a positive trend compared to the MPTP + vehicle treated group. Data are presented as mean + SEM (group 1: vehicle + vehicle, n 15; group 2: MPTP + vehicle, n 14; group 3: MPTP + compound 1(30mg/kg), n 13). Statistical significance: p <0.05, group 2: MPTP + vehicle with group 1: vehicle + vehicle (unpaired t-test).

Figure 37 reports the short-term effect of MPTP and compound 1 on T cell infiltration into the brain. The total number of CD 3-positive T cells counted in the substantia nigra of 3 30 μm thick sections from each mouse at study day 4 is presented. Data shown are mean ± s.e.m; p < 0.001; single factor analysis of variance, Sidak post hoc multiple comparison test. Mice treated with MPTP + compound 1 showed a tendency to have fewer CD3 positive cells, indicating a tendency for fewer T cells to infiltrate into the brain.

Figure 38 reports the short-term effects of MPTP and compound 1 on microglial proliferation at study day 4. Figure 38A depicts the extent of CD68 positive area measured in striatum of 3 30 μm thick sections from each mouse on study day 4. Fig. 38B depicts the extent of CD68 positive area measured in the substantia nigra compact of 3 30 μm thick sections from each mouse on study day 4. These data indicate that there is significant microglial proliferation in the substantia nigra (the major site of neuronal loss observed in parkinson's disease) but not in the striatum (the major brain region from neuronal projections from the substantia nigra), suggesting that treatment affects the central site of disease pathology without affecting the secondary projection region.

4. Synuclein transgenic mouse model for parkinson's disease

Synuclein transgenic mouse models were used to determine the effect of compound 1 on chronic CCR3 inhibition and its ability to slow, halt or reverse the progression of parkinson-like symptoms. This model of overexpression of human α -synuclein can test whether interventional therapy can prevent α -synuclein-induced behavioral and pathological effects. To select the best synuclein model for testing, plasma eotaxin levels were measured as biomarkers in multiple synuclein mouse models at different ages. These models include a53T, DxJ9M and Line 61. Plasma of 6-month-old Line 61 synuclein transgenic mice was measured by ELISA assay to detect plasma eotaxin levels as biomarkers. Line 61 mice aged 6 months (QPS Neuro, Grambach, Austria) showed transgenically induced increases in eotaxin levels (fig. 39), suggesting that plasma eotaxin levels may serve as appropriate clinical biomarkers to select treatment populations.

Thirty-three (33)4.5 month old male α -synuclein transgenic mice (Line 61) were divided into two groups (group a, n-17, compound 11 mg/mL; group B, n-16, vehicle) and a group of 15 non-transgenic age-matched littermates (group C, vehicle) were treated for 6 weeks with drinking water. Animals received vehicle or compound 1. The behavior of the animals was assessed at the end of the treatment period. Subsequently, tissue processing is performed.

Figure 40 reports the results from the wire hanging test. The average steel wire hang time for each group is shown, with animals from group C (non-transgenic, vehicle treated) exhibiting significantly higher steel wire hang times. Animals from group a (transgenic, compound 1-treated) exhibited significantly higher steel wire hang times than group B (transgenic, vehicle-treated). Data are shown as mean + SEM of all animals per group; p < 0.001; dunn post hoc inspection; mann Whitney test P <0.05 for group a versus group B. These data indicate that synuclein mice have a significant loss of their ability to hang on steel wire, which is at least partially rescued by compound 1 treatment.

Fig. 41 reports the results from the grip strength test. The average maximum grip g of each group is shown. Animals from groups a and C (transgenic, compound 1 treated and non-transgenic, respectively, vehicle treated) showed significantly higher grip strength compared to group B (transgenic, vehicle treated). Data are shown as mean ± SEM of all animals per group. Comparing each group to vehicle-treated transgenic animals (group B); one-way anova followed by Bonferroni post-test. These data indicate a statistically significant deficiency in synuclein mouse forelimb strength, which was completely rescued by compound 1 treatment.

Fig. 42 reports the results from the balance beam walk test. The number of animals per group that were able to pass completely through the balance beam in each trial is shown. No animals in group B (transgenic, vehicle treated) were able to cross the fifth (most difficult) balance bar, indicating that group a transgenic mice treated with compound 1 performed better than group B transgenic mice treated with vehicle and that the improved performance in the balance bar walk test was closer to the non-transgenic control.

Figure 43 reports the results of slippage or falls in each test for five balance-wood walking tests of three groups of mice (group a, transgenic treated with compound 1; group B, transgenic treated with vehicle; and group C, non-transgenic treated with vehicle). The average number of slips [ n ] for each group is shown, and each graph shows one test (1 to 5). Data are mean ± SEM of all animals per group. Comparing each group to group B; one-way anova followed by Bonferroni post hoc tests. These data clearly show that the ability of synuclein overexpressing mice to cross the balance bar is highly compromised and treatment with compound 1 at least partially rescues this effect, showing a significant improvement in motor function.

Figure 44 reports eosinophil counts from peripheral blood. Figure 44A reports the percentage of eosinophils in peripheral blood from three groups of mice (Tg Cmpd 1 ═ transgene treated with compound 1; Tg Veh ═ transgene treated with vehicle; nTG Veh ═ non-transgene treated with vehicle). The data were compared by t-test. Figure 44B reports the absolute eosinophil count in peripheral blood from three groups of mice (Tg Cmpd 1 ═ transgene treated with compound 1; Tg Veh ═ transgene treated with vehicle; nTG Veh ═ non-transgene treated with vehicle). The data were compared by t-test. These data indicate that the parkinsonian model of synuclein overexpression leads to eosinophilia depletion, which reverts to the levels of non-transgenic mice treated with compound 1, suggesting a beneficial immunomodulatory effect in this parkinsonian model. This also suggests that determining eosinophil levels in parkinson's disease patients treated with compound 1 can be a biomarker for the disease, including determining the level of progression, arrest or regression of the disease, as well as the therapeutic effect.

Figure 45 reports the effect of compound 1 on neuroinflammation. Figure 45A reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and the CD68 positive area quantified in hippocampus of transgenic, compound 1-treated mice (n ═ 14, 12, and 15, respectively). Figure 45B reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and the CD68 positive area quantified in the striatum of transgenic, compound 1-treated mice (n ═ 15, 11, and 16, respectively). Figure 45C reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and transgenic, Iba1 positive area quantified in hippocampus of compound 1-treated mice (n ═ 14, 13, and 16, respectively). Figure 45D reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and the quantified Iba1 positive area in the striatum of transgenic, compound 1-treated mice (n ═ 15, 11, and 16, respectively). Data are mean +/-s.e.m.; p < 0.05. Figure 45E reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and transgenic, GFAP-positive astrocytes quantified in hippocampus of compound 1-treated mice (n ═ 14, 13, and 15, respectively). Figure 45F reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and the quantified GFAP positive area in the striatum of transgenic, compound 1-treated mice (n ═ 15, 13, and 15, respectively). Data are mean +/-s.e.m.; p < 0.05. Figure 45G reports on non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and transgene, Iba-1 positive area quantified in substantia nigra dense part of compound 1 treated mice.

These data indicate that while overexpression of synuclein does not result in significant microglial hyperplasia as demonstrated by CD68 and Iba1 or astrocytosis as demonstrated by GFAP immunoreactivity, treatment with compound 1 reduced microglial hyperplasia of both markers and astrocytosis of GFAP, which may have an overall anti-inflammatory effect, i.e., not simply against one inflammatory cell type. Furthermore, the striatum is affected more than the hippocampus, suggesting that the region associated with parkinson's disease specifically reduces the microglial proliferation and astrocytic proliferation markers. This is further demonstrated by the effect of compound 1 on reversing microglial hyperplasia in the substantia nigra pars compacta.

Figure 46 reports compound 1 versus non-transgenic, vehicle-treated mice; transgenic, vehicle-treated mice; and effects of circulating levels of IL-4 and IL-6 cytokines in transgenic, compound 1-treated mice (n ═ 14, 15, and 17, respectively). Figure 46A reports the measured IL-4 levels in terminal heart plasma of all three groups. Figure 46B reports the measured IL-6 levels in terminal heart plasma of all three groups. Data shown are mean +/-s.e.m.; p < 0.05, P < 0.01; one-way anova, Dunnett's multiple comparison test after the event. Changes in these key cytokines indicate a general effect of compound 1 treatment on proteins involved in immune function and inflammation.

Infiltration of T cells into the brain

(a) Tissue processing and histology

Mice were sacrificed 2 hours after the last PO administration on the day 9 days after dosing. Anesthesia was induced by 2,2, 2-tribromoethanol. Mice were then perfused intracardiacally with a 1% EDTA in PBS followed by a 4% PFA in PBS. Brains were dissected and cut radially into uniform halves and fixed in 4% PFA in PBS. After two days of fixation, the brains were transferred to 30% sucrose in PBS and replaced two days later. Plasma was collected and stored on dry ice.

The half-brain was cut radially into 35 μm sections on a microtome at-22 ℃. Brain slices were collected into 12 tubes sequentially to represent every 12 th slice of the brain in a given tube. Brain sections were stored in cryoprotective media at-20 ℃ until staining was required. Free floating sections were blocked in the appropriate serum of 10% serum in PBST 0.5%. The primary antibody was incubated overnight at 4 ℃ or room temperature as described below.

1 at room temperature: 100 rat anti-CD 8a (63-0081-80, Thermo Fisher Scientific) was used at room temperature at a concentration of 1: 100 rat anti-CD 3(555273, BD Biosciences) was used at a concentration of 1: 1000 concentration Using rabbit anti-Iba-1 (016. sup. 26721, Wako) at 1: 1000 concentration rat anti-CD 68(MCA1957, Bio-Rad) was used, and the concentration was measured at 1: a concentration of 200 was used of Dylight 448/594-labeled tomato lectin (DL-1177, Fisher Scientific). The next day at room temperature 1: 300 concentration of a suitable fluorescent secondary antibody (Alexa-488/555/647, Invitrogen) was administered. The slides were covered using the Prolong Gold Mounting Media. Images were acquired at 20 x on a Hamamatsu nanozomer 2.0HT slide scanner.

(b) Quantification of T cell infiltration

CD3 and CD8 positive cells were counted from images taken from the cerebellum on a Hamamatsu slide scanner. The total number of CD3 and CD8 positive cells in the cerebellum was summed from about 5 35 μm sections. Statistical significance was tested using the general one-way anova and Dunnett's multiple post-hoc comparison test between treatment groups.

(c) Neuroinflammation quantification

Iba-1 and CD68 positive areas were quantified as a percentage of ROI around the entire cerebellum using a threshold on Image Pro Premier v9.2 software. About 5 sections from each mouse were averaged for Iba-1 and CD68 positive area percentage. Statistical significance was tested using the general one-way anova and Dunnett's multiple post-hoc comparison test between treatment groups.

Treatment group

Two month old C57Bl/6 mice were divided into three treatment groups: vehicle control, vehicle-treated EAE (Experimental autoimmune encephalomyelitis, see Methods Mol biol.2012; 900: 381-401, incorporated herein by reference in its entirety), and Compound 1-treated EAE. EAE was induced by Subcutaneous (SQ) MOG + CFA emulsion and Intravenous (IV) Pertussis Toxin (PT) on day 0. Additional PT injections were given on day 2. Vehicle or compound 1 administration was performed by oral gavage (PO) on day 0 and continued twice daily (BID) until day 9. Mice were removed 2 hours after the last PO administration.

Preparing the medicine: compound 1 was formulated in 40% HP- β -cyclodextrin and adjusted to pH 6.5 with NaOH (1M). The vehicle solution was similarly formulated and pH adjusted. Solutions were prepared fresh weekly and stored at 4 ℃.

EAE emulsion: MOG 35-55(Anaspec AS-60130-5) was reconstituted in PBS at 4 mg/ml. Mycobacterium tuberculosis H37 Ra (Fisher Scientific DF3114-33-8) was dissolved at 4mg/ml in incomplete Freud's adjuvant. The two solutions were then emulsified using a glass syringe (Thermo Scientific Male Luer-LOK printing Syringes 03-170-. Solutions were prepared just prior to administration.

Pertussis toxin: pertussis toxin (Sigma P7208-50UG) was dissolved at 0.002mg/ml in saline and injected at 100. mu.l IV on the day of induction and again two days later.

Treatment group:

treatment group 1, vehicle treatment: young C57BL/6 mice 2.5 months old (n ═ 5) received 100 μ l of vehicle PO BID for 9 days, starting 2 hours after SQ saline injection, with only 1 injection on the first and last day for a total of 19 treatment injections.

Treatment group 2, vehicle-treated EAE: young C57BL/6 mice 2.5 months old (n ═ 10) received 100 μ l of vehicle PO BID for 9 days, starting 2 hours after EAE induction, with only 1 injection on the first and last day for a total of 19 treatment injections.

Treatment group 3, compound 1 treated EAE: young C57BL/6 mice 2.5 months old (n ═ 10) received 100 μ l of compound 1(30mg/kg) PO BID for 9 days, starting 2 hours after EAE induction, with only 1 injection on the first and last day for a total of 19 treatment injections.

EAE induction resulted in an increase of CD3 and CD8 positive infiltrating T cells in the cerebellum. Treatment with compound 1 significantly reduced the number of these T cells. EAE induction also resulted in a significant increase in microglial proliferation in the cerebellum, which was also significantly rescued after 9 days of treatment with compound 1. Figure 47A shows an increase in CD3 positive infiltrating T cells in the cerebellum, which was significantly reduced after 9 days of treatment with compound 1. Figure 47B shows an increase in CD8 positive infiltrating T cells in the cerebellum, which was significantly reduced after 9 days of treatment with compound 1. FIG. 47C shows a significant increase in Iba-1 positive area in cerebellum after EAE, which was significantly reduced in cerebellum after 9 days of treatment. Figure 47D shows a significant increase in CD68 positive area in cerebellum after EAE, which was significantly reduced in cerebellum after 9 days of treatment.

d. Examples in humans

1. Eotaxin levels and senescence

The level of human eotaxin-1 was determined using a commercially available affinity-based assay (SOMAscan, SomaLogic, inc., Boulder, Colorado). Plasma samples were collected from 18, 30, 45, 55 and 66 year old donors and used for testing by SomaLogic using a SOMAscan aptamer-based affinity assay that tested, inter alia, for the relative levels of human eotaxin-1. Eotaxin-1 levels were determined and plotted against age group (FIG. 45). The relative eotaxin-1 concentrations increased with age, indicating a target for the eotaxin-1 pathway (including its primary receptor CCR3) to treat aging-related diseases such as neurodegenerative diseases and cognitive decline.

2. Human biomarker assays

Whole blood from compound 1-treated humans was incubated with human recombinant eotaxin-1 to trigger eosinophil shape change (fig. 48A) or CCR3 receptor internalization (fig. 49B). Both assays showed strong concentration-dependent effects of compound 1 on the corresponding functional biomarker readings.

In summary, the results obtained from the ESC and CCR3 receptor internalization assays demonstrate that compound 1 acts as an inhibitor of the human eotaxin-1 pathway. In particular, compound 1 may act as a potent inhibitor of CCR3 by binding to that receptor.

Furthermore, the data obtained in figure 44 show that eosinophil expression levels were lower in the transgenic parkinson's disease model in mammals than in non-transgenic animals, which were reversed after administration of compound 1. Thus, this data, which is applicable to parkinson's disease patients, can aid in the diagnosis, monitoring and determination of prognosis of the disease.

3. Treatment of subjects with parkinson's disease with compound 1

Subjects diagnosed with parkinson's disease were orally administered 400mg compound or placebo twice daily (BID). Subjects were treated for 12 weeks and then followed for 2 weeks of follow-up. The effect of compound 1 on motor function of subjects was evaluated under a practically defined non-drug treatment regime (which was greater than or equal to 12 hours without levodopa administration). Subjects were also evaluated by flow cytometry, pharmacogenomics, and biomarker analysis on blood and plasma samples, including eosinophil levels. Gait analysis was evaluated using Zeno walk. Bradykinesia, tremor, general activity and sleep were assessed using a wearable device.

Changes in baseline (day 1) motor function during the actual defined non-drug state at week 12 were determined using the Movement Disorder association's Unified Parkinson's Disease scoring Scale, MDS-UPDRS, section 3. Changes in baseline (day 1) clinical function, motor function and activities of daily living at week 12 during the dosing state were assessed by: MDS-UPDRS parts 1 to 4; montreal cognitive assessment (MoCA); schwab and England Activities of Daily Living, SE-ADL) scale; severity Index of Clinical Impression-PD (Clinical Impression of Severity Index, CISI-PD); PD Quality of Life Questionnaire-39 (PD Quality of Life questonaire-39, PDQ-39); the Sheehan suicide tendency Tracking Scale (S-STS); and 10 meter timed walking (which was also assessed in the non-drug state).

It is to be understood that the invention is not limited to the specific aspects described and as such may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

Where a range is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. When the range includes one or both of the limits, ranges that do not include one or both of those are also included in the invention.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, representative illustrative methods and materials are now described.

All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference, and are herein incorporated by reference to disclose and describe the methods and/or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

It should be noted that, as used herein and in the appended claims, no element preceding a quantity is intended to include the plural form unless the context clearly indicates otherwise. It is also noted that the claims can be drafted to exclude any optional element. Accordingly, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely," "only," and the like in connection with the recitation of claim elements, or use of a "negative" limitation.

As will be apparent to those of skill in the art upon reading this disclosure, each of the individual aspects described and illustrated herein has discrete components and features that can be readily separated from or combined with the features of any of the other several aspects without departing from the spirit and scope of the present disclosure. Any of the methods described may be performed in the order of events described or in any other order that is logically possible.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

Accordingly, the foregoing merely illustrates the principles of the invention. It will thus be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of the invention and the concepts contributed by the inventor to furthering the art, and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and aspects of the invention, as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof.

Additionally, it is intended that such equivalents include both currently known equivalents as well as equivalents developed in the future, i.e., any components developed that perform the same function, regardless of structure. Accordingly, the scope of the present invention is not intended to be limited to the exemplary aspects shown and described herein. Rather, the scope and spirit of the present invention is embodied by the appended claims.