Novel 5-hydroxypyridines as P2X1 and P2X3 receptor antagonists and pharmaceutical compositions containing the same
阅读说明：本技术 用作p2x1及p2x3受体拮抗剂的新型5-羟基吡啶类化合物及包含其的药物组合物 (Novel 5-hydroxypyridines as P2X1 and P2X3 receptor antagonists and pharmaceutical compositions containing the same ) 是由 金容哲 郑瑛焕 于 2018-03-13 设计创作，主要内容包括：本发明涉及用作P2X1和P2X3受体拮抗剂的新型5-羟基吡啶类化合物和包含该化合物的组合物。本发明的化合物具有强烈拮抗P2X1和P2X3受体的活性,因此可以有效地用作治疗或预防由P2X1和P2X3受体活性引起的慢性炎症性疾病或神经性疼痛疾病的药物。(The present invention relates to novel 5-hydroxypyridines useful as P2X1 and P2X3 receptor antagonists and compositions containing them. The compound of the present invention has strong antagonistic activity against P2X1 and P2X3 receptors, and thus can be effectively used as a drug for treating or preventing chronic inflammatory diseases or neuropathic pain diseases caused by P2X1 and P2X3 receptor activity.)
1. A compound represented by the following general formula 1, an isomer of the compound, or a pharmaceutically acceptable salt,
[ general formula 1 ]
In the case of the above-mentioned general formula 1,
a is any one of substituents represented by the following chemical formula 1 or chemical formula 2,
[ chemical formula 1 ]
[ chemical formula 2 ]
Wherein n and m are each independently an integer of 0 to 5;
R1is a hydrogen atom, a linear C which is unsubstituted or substituted by carboxyl, sulfonyl, thioether, benzyl, phenethyl1-C6Alkyl or branched C1-C6An alkyl group;
R2is hydrogen, unsubstituted or substituted by carboxyl, sulfonyl, cyano, amide, tetrazolyl or
R3is a hydrogen atom, unsubstituted or substituted by a carboxyl groupRadical-substituted straight-chain C1-C6Alkyl or branched C1-C6Alkyl, or with R2Together said 5-6 membered heterocyclyl;
R4is a hydrogen atom, or a linear C1-C6Alkyl or branched C1-C6An alkyl group;
R5is unsubstituted or straight-chain C substituted by phenyl, pyrrolidinyl, piperidinyl, tetrahydrofuran or 1, 3-dioxo-1, 3-dihydro-2H-isoindol-2-yl1-C8Alkyl or branched C1-C8Alkyl radical, C5-C6Cycloalkyl radicals or
2. The compound according to claim 1, an isomer of the compound, or a pharmaceutically acceptable salt thereof, when A of the compound represented by the general formula I is the chemical formula 1,
n is an integer of 0 to 5;
R1is a hydrogen atom, straight chain type C1-C4Alkyl or branched C1-C4Alkyl, carboxyl, -CH2CH2-S(＝O)2-CH3，-CH2CH2SCH3Benzyl or phenethyl;
R2is a hydrogen atom, C1-C3Alkyl, -COOH, -CH ═ CHCOOH, -CH2CH2COOH、-S(＝O)2-CH3Cyano, -CONH2Or
R3is a hydrogen atom, C1-C3Alkyl or carboxyl, or with R2Together form the 5-6 membered heterocyclyl;
R4is a hydrogen atom or C1-C3An alkyl group.
3. The compound, the isomer of the compound, or the pharmaceutically acceptable salt according to claim 1, when A of the compound represented by the general formula I is chemical formula 2,
m is an integer of 0 to 5;
R1is a hydrogen atom or a straight chain type C1-C3Alkyl or branched C1-C3An alkyl group;
R2is a hydrogen atom, straight chain type C1-C3Alkyl or branched C1-C3Alkyl, -COOH, -CH ═ CHCOOH, -CH2CH2COOH、-CH2NH2、-CH2NH-S(＝O)2-CH3Cyano or
R3Is a hydrogen atom, straight chain type C1-C3Alkyl or branched C1-C3Alkyl or-COOH;
R5is that
4. A5-hydroxypyridine compound, an isomer of the 5-hydroxypyridine compound, or a pharmaceutically acceptable salt thereof, wherein the 5-hydroxypyridine compound is represented by the following general formula Ia,
[ formula Ia ]
In the general formula Ia described above, the,
R1is a hydrogen atom, straight chain type C1-C4Alkyl or branched C1-C4Alkyl, -COOH, -CH2CH2-S(＝O)2-CH3、-CH2CH2SCH3Benzyl or phenethyl;
R2is a hydrogen atom, C1-C3Alkyl, -COOH, -CH ═ CHCOOH, -CH2CH2COOH、-S(＝O)2-CH3Cyano, -CONH2Or
R3is a hydrogen atom, C1-C3Alkyl or carboxyl, or with R2Together form the 5-6 membered heterocyclyl;
R4is a hydrogen atom or C1-C3An alkyl group.
5. A5-hydroxypyridine compound, an isomer of the 5-hydroxypyridine compound or a pharmaceutically acceptable salt thereof, wherein the 5-hydroxypyridine compound is represented by the following general formula Ib,
[ formula Ib ]
In the general formula Ib, R1Is a hydrogen atom or a straight chain type C1-C3Alkyl or branched C1-C3An alkyl group;
R2is a hydrogen atom, a straight chain or a branched chain C1-C3Alkyl, -COOH, -CH ═ CHCOOH, -CH2CH2COOH、-CH2NH2、-CH2NH-S(＝O)2-CH3Cyano or
R3Is a hydrogen atom, a straight chain or a branched chain C1-C3Alkyl or-COOH;
R5is that
6. The 5-hydroxypyridine compound of claim 5, an isomer of the 5-hydroxypyridine compound, or a pharmaceutically acceptable salt thereof, wherein the compound is represented by the following general formula Ib',
[ formula Ib' ]
In the formula Ib', R1Is a hydrogen atom or a straight chain type C1-C3Alkyl or branched C1-C3An alkyl group;
R3is a hydrogen atom or a straight chain type C1-C3Alkyl or branched C1-C3An alkyl group;
R5is that
7. The compound, an isomer of the compound, or a pharmaceutically acceptable salt of the compound according to claim 1, wherein the compound represented by formula 1 is any one selected from the following group of compounds:
5-hydroxy-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 a);
5-hydroxy-2- (4-methoxyphenethyl) -6-methylpyridine-3, 4-dicarboxylic acid (13 b);
6-ethyl-5-hydroxy-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 c);
5-hydroxy-2- (4-methoxyphenethyl) -6-propylpyridine-3, 4-dicarboxylic acid (13 d);
6-butyl-5-hydroxy-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 e);
5-hydroxy-6-isopropyl-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 f);
5-hydroxy-6-isobutyl-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 g);
6-benzyl-5-hydroxy-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 h);
5-hydroxy-2- (4-methoxyphenethyl) -6-phenethylpyridine-3, 4-dicarboxylic acid (13 i);
3-hydroxy-6- (4-methoxyphenethyl) pyridine-2,4, 5-tricarboxylic acid (13 j);
5-hydroxy-2- (4-methoxyphenylethyl) -6- (2- (methylthioethyl) ethyl) pyridine-3, 4-dicarboxylic acid (13 k);
5-hydroxy-2- (4-methoxyphenylethyl) -6- (2- (methylsulfonyl) ethyl) pyridine-3, 4-dicarboxylic acid (13 l);
4-cyano-5-hydroxy-2- (4-methoxyphenylethyl) -6-methylnicotinic acid ethyl ester (14);
ethyl 5-hydroxy-2- (4-methoxyphenylethyl) -6-methyl-4- (H-tetrazol-5-yl) nicotinate (15);
4-cyano-5-hydroxy-2- (4-methoxyphenylethyl) -6-methylnicotinic acid (16);
5-hydroxy-2- (4-methoxyphenylethyl) -6-methyl-4- (H-tetrazol-5-yl) nicotinic acid (17);
7-hydroxy-4- (4-methoxyphenylethyl) -6-methylfuran [3,4-c ] pyridine-1, 3-dione (18);
8-hydroxy-5- (4-methoxyphenethyl) -7-methyl-2, 3-dihydropyrido [3,4-d ] pyridazine-1, 4-dione (19);
6- (4-methoxyphenylethyl) -2-methyl-4- (methylsulfonyl) pyridin-3-ol (20);
3-hydroxy-6- (4-methoxyphenylethyl) -2-methylisonicotinic acid methyl ester (21 a);
3-hydroxy-6- (4-methoxyphenylethyl) -2, 5-dimethylisonicotinic acid methyl ester (21 b);
3-hydroxy-6- (4-methoxyphenylethyl) -5-methyl-2-propylisonicotinic acid methyl ester (21 c);
3-hydroxy-6- (4-methoxyphenylethyl) -2-methylisonicotinic acid (22 a);
3-hydroxy-6- (4-methoxyphenylethyl) -2, 5-dimethylisonicotinic acid (22 b);
3-hydroxy-6- (4-methoxyphenylethyl) -5-methyl-2-propylisonicotinic acid (22 c); and
5-hydroxy-2- [2- (2- (4-methoxyphenyl) ethyl ] -3-methyl-6-propyl-pyridine-4-carboxamide (23);
5-hydroxy-2- (3-phenoxybenzyl) -6-propylpyridine-3, 4-dicarboxylic acid dimethyl ester (26);
5-hydroxy-2- (3-phenoxybenzyl) -6-propylpyridine-3, 4-dicarboxylic acid (27);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid methyl ester (28);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid (29);
3- (benzyloxy) -5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid methyl ester (30);
(3- (benzyloxy) -5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) methanol (31);
4- (hydroxymethyl) -5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-3-ol (32);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinal (33);
(E) -ethyl 3- (3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) acrylate (34);
(E) -3- (3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) acrylic acid (35);
ethyl3- (3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) propionate (36);
3- (3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) propionic acid (37)
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid nitrile (38);
3- (benzyloxy) -5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid nitrile (39);
(3- (cyclohexa-2, 4-dien-1-ylmethoxy) -5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) methylamine (40);
4- (aminomethyl) -5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-3-ol (41);
n- ((3- (benzyloxy) -5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) methyl) methanesulfonamide (42);
n- ((3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) methyl) methanesulfonamide (43);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid phenethyl ester (46 a);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid octyl ester (46 b);
2-phthalimidoethyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (46 c);
4-phthalimidobutyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (46 d);
2- (pyrrolidin-1-yl) ethyl3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (46 e);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid isopropyl ester (46 f);
pent-3-yl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (46 g);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid cyclopentyl ester (46 h);
tetrahydrofuran-3-yl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (46 i);
1- (tert-butoxycarbonyl) piperidin-4-yl3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (46 j);
piperidin-4-yl3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (47); and
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid ethyl ester (48).
8. A pharmaceutical composition for preventing or treating a chronic inflammatory disease, a neuropathic pain disease, or a platelet aggregation-related disease, comprising the compound, an isomer of the compound, or a pharmaceutically acceptable salt of the compound according to any one of claims 1 to 7 as an active ingredient.
9. The pharmaceutical composition of claim 8, wherein the chronic inflammatory disease is degenerative arthritis, rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, or cystitis.
10. The pharmaceutical composition according to claim 8, wherein the neuropathic pain disorder is neuropathic pain, allodynia, diabetic neuropathy, idiopathic pain, irritative pain, phantom limb pain, or complex regional pain syndrome.
11. The pharmaceutical composition according to claim 8, wherein the disease associated with platelet aggregation is arteriosclerosis, stroke, thrombosis, embolism, myocardial infarction, atherosclerosis or peripheral blood circulation disorder.
12. An antagonist of the P2X receptor, comprising the compound of any one of claims 1 to 7, an isomer of the compound, or a pharmaceutically acceptable salt thereof as an active ingredient.
Technical Field
The present invention was made with the support of the future department of creative sciences by the project No. 2014R1A2A1a11052300, the research administration and management professional organization of which is korean research foundation, the research industry name of "core personal research (achievement diffusion)", the research project name of "research on multi-target control mechanism of neuropathic pain signaling and drug design", the competent organization of which is korean science and technology institute; the duration of the study was 2014.11.01-2016.10.31.
The present patent application claims priority of korean patent application No. 10-2017-0031413, filed on 13.3.2017 to the korean intellectual property office, and the disclosure of the above patent application is incorporated into the present specification by reference.
The present invention relates to novel 5-hydroxypyridines as P2X1 and P2X3 receptor antagonists and pharmaceutical compositions containing them.
Background
The P2X receptor is a ligand-gated cation channel that is activated in response to binding of extracellular adenosine 5' -triphosphate (ATP). It is well known that when the transmembrane pore (transmembrane pore) of the P2X receptor is opened, such as Ca2+，Na+And K+Enter the cytoplasm, causing membrane depolarization and cellular excitability. In mammals, the seven P2X receptor subtypes are widely expressed in vascular smooth muscle, platelets, the peripheral and central nervous systems, and immune cells, and mediate a variety of physiological processes, including synaptic transmission, presynaptic mediation, smooth muscle contraction, cell proliferation and cell death, visceral motility, platelet aggregation, taste, pain sensation, and inflammation. Recent studies of the X-ray crystal structure of the zebrafish P2X4 receptor have shown that the P2X receptor has a common topology, including two transmembrane domains (TM1 and TM2), an ATP binding site, and a large glycosylated and cysteine-rich extracellular loop containing intracellular N-and C-termini. Also, it is known that the gating mechanism of ATP binding to zP2X4 receptor is determined by the steric structural change between the apolipoprotein (apo) state and the open (open) state of the crystal structureAnd (4) determining.
Among the P2X receptor subtypes, the P2X3 receptor (P2X3R) is present in C-and a δ -fiber primary afferent neurons of small and medium diameter in the form of homo (homomeric) P2X3 and hetero (heterotomeric) P2X2/3 receptors, which provides a high degree of specificity for the pain-sensing system a number of studies including knockout animal studies (knock-out animal studies) indicate that P2X3R is closely associated with pain perception, for example, administration of ATP or P2X 3R-selective agonist (agonst) αβ -methylene ATP to rat hindpaw causes pain, in neuropathic or inflammatory pain animal models, knock-out P2X3R or animals using P2X3R selective antisense RNA or siRNA (short interfering RNA) have a significantly reduced pain-associated behaviour in animals, furthermore, treatment with P2X4 receptor antagonists TNP-ATP (2',3' -O2, 3' -O) or siRNA (short interfering RNA) has shown that the potential for pain control of the effects of P2X R on P-and P2X 3-nitrophenyl responses in these animal models.
Over the past decade, potent and selective P2X3 receptor antagonists have been developed by the pharmaceutical industry and academic institutions, including the present inventors. Yapei Laboratories (Abbott Laboratories) disclosed A-317491 (Compound 1 of FIG. 1a), which is the first selective non-nucleotide dual (dual) P2X3-P2X2/3 antagonist, shown to reduce pain in chronic and inflammatory pain animal models. Roche (Roche) reported analogs RO-4 (Compound 2a of FIG. 1b), RO-51 (Compound 2b of FIG. 1b), and RO-85 (Compound 3 of FIG. 1c) of the diaminopyrimidine (diaminopyrimidine) class as the first allosteric (alloteric) antagonist with high potency and selectivity for the P2X3-P2X2/3 receptor. Astrazepam (AstraZeneca) to improve the efficacy and physicochemical properties of P2X3 receptor antagonists, a study of the structure-activity relationship (SAR) of pyrrolopyrimidinone (pyridone) derivatives (compound 4 of fig. 1 d) was published. An oral selective P2X3-P2X2/3 receptor antagonist AF-219 with strong biocompatibility as a clinical candidate drug of affinity pharmaceutical company (affinity Pharmaceuticals) due to the inclusion of interstitial cystitis/bladderResults of several proof-of-concept studies in phase II clinical trials of cystalgia syndrome and chronic cough show significant pharmacological effects. The present inventors reported that the selective and non-competitive peptide antagonist silibinol (Spinorphin) (compound 5, IC of figure 1 e)508.3pM) and 5-hydroxypyridine derivatives (compounds 7a-c of fig. 1 g), and these compounds were developed by transforming the strong anionic phosphosulfonic acid group and the unstable azo bond (-N ═ N-) of PPADS (compound 6a in fig. 1 f) as a non-selective P2X antagonist into a weak anionic carboxylic acid and a stable carbon-carbon bond, respectively.
In order to optimize the drug properties of 5-hydroxypyridine derivatives (compounds 7a-c of fig. 1 g), the present inventors introduced various substituents at various positions of carbon, and as a result, developed novel hP2X3 receptor antagonists having higher biological stability and excellent antagonistic action.
Throughout this specification, reference is made to a number of papers and patent documents, the contents of which are shown. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to more specifically describe the state of the art to which this invention pertains and the contents of this invention.
Disclosure of Invention
Problems to be solved by the invention
The present inventors have attempted to develop a novel antagonist against the P2X1 receptor and the P2X3 receptor, which play an important role in pain and inflammatory signaling. The present inventors synthesized novel compounds (compounds 7a-c of fig. 1 g) having various substituents introduced to carbons at various positions of 5-hydroxypyridine derivatives, and tested the results of pharmacokinetics and efficacy thereof, and completed the present invention by elucidating hP2X3 receptor antagonists which have higher biostability and excellent antagonistic action.
Therefore, the present invention aims to provide novel 5-hydroxypyridines useful as P2X1 and P2X3 receptor antagonists, and pharmaceutical compositions comprising the same.
Another object of the present invention is to provide a method for treating chronic inflammatory diseases, neuropathic pain diseases or platelet aggregation-related diseases, in a step of administering to a subject a pharmaceutical composition comprising the above novel 5-hydroxypyridine compounds serving as the P2X1 receptor and the P2X3 receptor.
Other objects and advantages of the present invention will become more fully apparent from the following detailed description of the invention, the appended claims and the accompanying drawings.
Means for solving the problems
According to an aspect of the present invention, there is provided a compound represented by the following general formula 1, an isomer of the above compound, or a pharmaceutically acceptable salt thereof:
[ general formula 1 ]
In the above-mentioned general formula 1,
a is any one of substituents represented by the following chemical formula 1 or 2,
[ chemical formula 1 ]
[ chemical formula 2 ]
Wherein n and m are each independently an integer of 0 to 5;
R1is a hydrogen atom, unsubstituted or substituted by carboxyl, sulfonyl, thioether (e.g. -CH)2CH2SCH3) Benzyl, phenethyl substituted straight or branched C1-C6An alkyl group;
R2is hydrogen, unsubstituted or substituted by carboxyl, sulfonyl, cyano, amide, tetrazolyl or
R3is a hydrogen atom, a linear or branched C which is unsubstituted or substituted by a carboxyl group1-C6Alkyl, or with R2Together form the 5-to 6-membered heterocyclyl described above;
R4is a hydrogen atom, or a linear or branched C1-C6An alkyl group;
R5is unsubstituted or substituted by phenyl, pyrrolidinyl, piperidinyl, tetrahydrofuran or 1, 3-dioxo-1, 3-dihydro-2H-isoindol-2-yl1-C8Alkyl radical, C5-C6Cycloalkyl radicals or
According to one embodiment of the present invention, R of formula I above2May be a hydrogen atom, C1-C6Alkyl, -COOH, -CH ═ CHCOOH, -CH2CH2COOH, methylsulfonyl (-S (═ O)2-CH3) Cyano, amides (-CONH)2)、
Wherein R is5Can be
When a of the compounds represented by the above general formula I is the above chemical formula 1, the group of compounds belonging to the above general formula I includes the group of compounds having the following characteristics:
n is an integer of 0 to 5; r1Is a hydrogen atom, unsubstituted or substituted by a carboxyl group, a sulfonyl group,Thioethers (e.g. -CH)2CH2SCH3) Benzyl, phenethyl substituted straight or branched C1-C4An alkyl group; r2Is hydrogen, unsubstituted or substituted by carboxyl, sulfonyl, cyano, amide, tetrazolyl or
And, when a of the compounds represented by the above general formula I is the above chemical formula 2, the group of compounds belonging to the above general formula I includes the group of compounds having the following characteristics:
m is an integer of 0 to 5; r1Is a hydrogen atom, or a straight or branched C1-C3An alkyl group; r2Is a hydrogen atom, unsubstituted or substituted by carboxyl, -CH2NH2、-CH2NH-S(＝O)2-CH3Cyano or
Further, according to an embodiment of the present invention, there is provided a 5-hydroxypyridine compound represented by the following general formula Ia, an isomer of the compound, or a pharmaceutically acceptable salt of the compound:
[ formula Ia ]
In the above-mentioned general formula Ia,
R1is a hydrogen atom, a straight chain or a branched chain C1-C4Alkyl, carboxyl (-COOH), methylsulfonylethyl (-CH)2CH2-S(＝O)2-CH3)、-CH2CH2SCH3Benzyl or phenethyl; r2 is a hydrogen atom, C1-C3Alkyl, -COOH, -CH ═ CHCOOH, -CH2CH2COOH, methylsulfonyl (-S (═ O)2-CH3) Cyano, amides (-CONH)2) OrOr with R3Together form a five-to six-membered heterocyclic group containing at least one atom selected from the group consisting of N, O, S; r3Is a hydrogen atom, C1-C3Alkyl or carboxyl, or with R2Together form a 5-6 membered heterocyclyl. R4Is a hydrogen atom or C1-C3An alkyl group.
In addition, according to another embodiment of the present invention, there is provided a 5-hydroxypyridine compound represented by the following general formula Ib:
[ general formula Ib ]
In the above general formula Ib, R1Is a hydrogen atomSub-or straight-chain or branched C1-C3An alkyl group; r2Is a hydrogen atom, a straight chain or a branched chain C1-C3Alkyl, -COOH, -CH ═ CHCOOH, -CH2CH2COOH、-CH2NH2Sulfonyl group (-CH)2NH-S(＝O)2-CH3) Cyano or
Further, according to still another embodiment of the present invention, there is provided a 5-hydroxypyridine compound represented by the following general formula Ib', an isomer thereof, or a pharmaceutically acceptable salt thereof:
[ formula Ib' ]
In the general formula Ib',
R1is a hydrogen atom or a straight or branched C1-C3An alkyl group; r3Is a hydrogen atom or a straight or branched C1-C3An alkyl group; r5Is that
Among the compounds belonging to the above formula Ia, there may be included:
5-hydroxy-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 a);
5-hydroxy-2- (4-methoxyphenethyl) -6-methylpyridine-3, 4-dicarboxylic acid (13 b);
6-ethyl-5-hydroxy-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 c);
5-hydroxy-2- (4-methoxyphenethyl) -6-propylpyridine-3, 4-dicarboxylic acid (13 d);
6-butyl-5-hydroxy-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 e);
5-hydroxy-6-isopropyl-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 f);
5-hydroxy-6-isobutyl-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 g);
6-benzyl-5-hydroxy-2- (4-methoxyphenethyl) pyridine-3, 4-dicarboxylic acid (13 h);
5-hydroxy-2- (4-methoxyphenethyl) -6-phenethylpyridine-3, 4-dicarboxylic acid (13 i);
3-hydroxy-6- (4-methoxyphenethyl) pyridine-2,4, 5-tricarboxylic acid (13 j);
5-hydroxy-2- (4-methoxyphenylethyl) -6- (2- (methylthioethyl) ethyl) pyridine-3, 4-dicarboxylic acid (13 k);
5-hydroxy-2- (4-methoxyphenylethyl) -6- (2- (methylsulfonyl) ethyl) pyridine-3, 4-dicarboxylic acid (13 l);
4-cyano-5-hydroxy-2- (4-methoxyphenylethyl) -6-methylnicotinic acid ethyl ester (14);
ethyl 5-hydroxy-2- (4-methoxyphenylethyl) -6-methyl-4- (H-tetrazol-5-yl) nicotinate (15);
4-cyano-5-hydroxy-2- (4-methoxyphenylethyl) -6-methylnicotinic acid (16);
5-hydroxy-2- (4-methoxyphenylethyl) -6-methyl-4- (H-tetrazol-5-yl) nicotinic acid (17);
7-hydroxy-4- (4-methoxyphenylethyl) -6-methylfuran [3,4-c ] pyridine-1, 3-dione (18);
8-hydroxy-5- (4-methoxyphenethyl) -7-methyl-2, 3-dihydropyrido [3,4-d ] pyridazine-1, 4-dione (19);
6- (4-methoxyphenylethyl) -2-methyl-4- (methylsulfonyl) pyridin-3-ol (20);
3-hydroxy-6- (4-methoxyphenylethyl) -2-methylisonicotinic acid methyl ester (21 a);
3-hydroxy-6- (4-methoxyphenylethyl) -2, 5-dimethylisonicotinic acid methyl ester (21 b);
3-hydroxy-6- (4-methoxyphenylethyl) -5-methyl-2-propylisonicotinic acid methyl ester (21 c);
3-hydroxy-6- (4-methoxyphenylethyl) -2-methylisonicotinic acid (22 a);
3-hydroxy-6- (4-methoxyphenylethyl) -2, 5-dimethylisonicotinic acid (22 b);
3-hydroxy-6- (4-methoxyphenylethyl) -5-methyl-2-propylisonicotinic acid (22 c); and
5-hydroxy-2- [2- (2- (4-methoxyphenyl) ethyl ] -3-methyl-6-propyl-pyridine-4-carboxamide (23).
Compounds belonging to the above formula Ib or Ib' may include:
5-hydroxy-2- (3-phenoxybenzyl) -6-propylpyridine-3, 4-dicarboxylic acid dimethyl ester (26);
5-hydroxy-2- (3-phenoxybenzyl) -6-propylpyridine-3, 4-dicarboxylic acid (27);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid methyl ester (28);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid (29);
3- (benzyloxy) -5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid methyl ester (30);
(3- (benzyloxy) -5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) methanol (31);
4- (hydroxymethyl) -5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-3-ol (32);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinal (33);
(E) -ethyl 3- (3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) acrylate (34);
(E) -3- (3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) acrylic acid (35);
ethyl3- (3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) propionate (36);
3- (3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) propionic acid (37)
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid nitrile (38);
3- (benzyloxy) -5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid nitrile (39);
(3- (cyclohexa-2, 4-dien-1-ylmethoxy) -5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) methylamine (40);
4- (aminomethyl) -5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-3-ol (41);
n- ((3- (benzyloxy) -5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) methyl) methanesulfonamide (42);
n- ((3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylpyridin-4-yl) methyl) methanesulfonamide (43);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid Phenethyl ester (phenyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinite) (46 a);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid Octyl ester (Octyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinite) (46 b);
2-phthalimidoethyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (2-phthalimidoethyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate) (46 c);
4-phthalimidobutyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (4-phthalimidobutyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate) (46 d);
2- (pyrrolidin-1-yl) ethyl3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (2- (pyrrolidin-1-yl) ethyl3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate) (46 e);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid Isopropyl ester (isoproyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinite) (46 f);
pent-3-yl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (Pentan-3-yl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate) (46 g);
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid Cyclopentyl ester (Cyclopentyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinite) (46 h);
tetrahydrofuran-3-yl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (Tetrahydrofuran-3-yl 3-hydroxy-5-methyl-6- (3-phenyloxybenzyl) -2-propylisonicotinate) (46 i);
1- (Tert-butoxycarbonyl) piperidin-4-yl3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (1- (Tert-butoxycarbonyl) piperidine-4-yl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate) (46 j);
piperidin-4-yl3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate (Piperidin-4-yl3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinate) (47); and
3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid Ethyl ester (Ethyl 3-hydroxy-5-methyl-6- (3-phenoxybenzyl) -2-propylisonicotinic acid) (48).
The compounds of the present invention represented by the above general formula I can be prepared into pharmaceutically acceptable salts and solvates according to conventional methods in the art.
Acid addition salts formed from pharmaceutically acceptable free acids (free acids) are useful as salts. Acid addition salts may be prepared by conventional methods, for example, by dissolving the compound in an excess of aqueous acid and precipitating the salt by using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. An acid or an alcohol (for example, ethylene glycol monomethyl ether) in an amount equimolar to the compound and water may be heated, and then the mixture may be dried by evaporation, or the precipitated salt may be suction-filtered.
In this case, as the free acid, an organic acid or an inorganic acid can be used. As the inorganic acid, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, or the like; as the organic acid, methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid (propionic acid), citric acid (citricacid), lactic acid (lactic acid), glycolic acid (glycolic acid), gluconic acid (gluconic acid), galacturonic acid, glutamic acid, glutaric acid (glutaric acid), glucuronic acid (glucoronic acid), aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, and the like can be used.
Also, bases may be used to prepare pharmaceutically acceptable metal salts. For example, the alkali metal salt or alkaline earth metal salt is obtained by dissolving the compound in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, and filtering the undissolved salt of the compound, followed by evaporating and drying the filtrate. Here, as the metal salt, it is most suitable to prepare a sodium salt, a potassium salt or a calcium salt in terms of pharmaceutical formulation, and a silver salt corresponding to the metal salt can be obtained by reacting an alkali metal or alkaline earth metal salt with an appropriate silver salt (for example, silver nitrate).
Unless otherwise indicated, pharmaceutically acceptable salts of 5-hydroxypyridines having the structure of formula I above may include salts of acidic or basic groups that may be present in 5-hydroxypyridines having the structure of formula I. For example, pharmaceutically acceptable salts may include sodium, calcium and potassium salts of hydroxyl groups, and other pharmaceutically acceptable salts of amino groups may include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, mesylate (tosylate), tosylate (tosylate), and the like, which salts may be prepared by preparation methods known in the art or prepared too long.
In addition, the 5-hydroxypyridine compounds having the structure of formula I have asymmetric centers and thus may exist in different enantiomeric forms, and all optical isomers and R or S type stereoisomers of the 5-hydroxypyridine compounds having the structure of formula I and mixtures thereof are within the scope of the present invention. The present invention includes the use of racemates, more than one enantiomeric form, more than one diastereomeric form or mixtures thereof, and also includes separation or preparation methods of the isomers as known in the art.
It is another object of the present invention to provide a method for preparing the above compound of formula I, which can be chemically synthesized by the method shown in the following reaction scheme, but is not limited to these examples.
The following reaction formulae show the preparation method of a representative compound of the present invention in terms of preparation steps, and the present invention is prepared by minor modifications (modification of reagents, solvents and reaction order used in the synthesis process of reaction formulae 1 to 8).
Some of the compounds of the present invention were synthesized according to procedures not within the scope of the equations, and detailed synthetic methods for these compounds are described in their respective examples.
(reaction formula 1)3, 4-dicarboxylpyridine derivative (C)3,4-Dicarboxypyridine derivatives) Preparation of
Reagents and conditions in the above reaction scheme 1: (a) EDC, TEA, DCM, RT, 2h, 55-99%; (b) p2O5，CHCl3Refluxing for 5h, 58-99%; (b') triphenylphosphine, iodine, TEA, DCM, RT, 12h, 50-63%; (c) dimethyl maleate, pure (neat), reflux, 5h, 21-52%; (d)
(reaction formula 2) 5-hydroxyPhenylpyridine derivatives (A)5-Hydroxypyridine Derivatives) Preparation of (Compound 16-19)
(reaction formula 2-1)
(reaction formula 2-2)
(reaction formula 2-3)
Reagents and conditions of equation 2(2-1 to 2-3) are (a) cis- β -cyanoacrylate, neat (neat), reflux, 5h, 27-55%, (b) sodium azide, ammonium chloride, DMF, 90 ℃, 12h, 48-62%, (c) 20% KOH (aq), RT, 6h, 30-52%, (d) maleic anhydride, benzene, 60 ℃, 2h, 43-54%, (e) hydrazine hydrate (aq), anhydrous sodium acetate, acetic acid, reflux, 2h, 35-46%, (f) methyl vinyl sulfone, neat, reflux, 5h, 49-55%.
(reaction formula 3) preparation of 5-hydroxypyridine derivative (Compound 21-23)
Reagents and conditions in equation 3 above: (a) methyl acrylate, pure (neat), 5h, 37% -46%; (a') methyl crotonate, neat (neat), 5h, 32% -52%; (b) 20% KOH (aq), RT, 6h, 40% -57%; (c) 30% ammonia water (aq), RT, 12h, 32% -45%.
(reaction formula 4) preparation of 5-Hydroxypyridine derivative (5-Hydroxypyridine Derivatives) (Compounds 36-39, 33-37)
(reaction formula 4-1)
(reaction formula 4-2)
Reagents and conditions of equation 4: (a) p2O5，CHCl3Refluxing for 5h, 85% -99%; (b) methyl crotonate, pure and refluxed for 5 hours, 22 to 52 percent; (c) benzyl bromide, K2CO3Acetone, refluxing for 2h, 73-96%; (d) LAH, ether, 0 ℃, 1h, 67-88%; (e) Pd/C, H2，RT，0.5h，54％-91％；(f)MnO2DCM, RT, 2h, 66-85%; (g) NaH, triethyl phosphonoacetate, THF, RT, 1h, 63% -78%; (h)20 percent of KOH (aq), room temperature, 6h, 37 percent to 94 percent; (i) dimethyl maleate, pure, refluxing, 5h, 32% -54%.
(reaction formula 5) preparation of derivative of 5-Hydroxypyridine (5-Hydroxypyridine Derivatives) (Compounds 38-43)
Reagents and conditions of equation 5: (a) methyl crotononitrile, pure, refluxing, 5h, 52% -74%; (b) benzyl bromide, K2CO3Acetone, refluxing for 2h, 62-86%; (c) LAH, ether, 0 ℃, 1h, 52-74%; (d) methanesulfonyl chloride, TEA, DCM, RT, 1h, 61% -77%; (e) Pd/C, H2, RT, 0.5H, 60% -91%.
(reaction formula 6) preparation of Compound 46a-j
Reagents and conditions of equation 6: (a) 20% of KOH, methanol, 1, 4-dioxane, 50 ℃, 2d, 63%; (b) alkyl bromides, NaHCO3，DMF，60℃，5h，35％-89％；(c)Pd/C，H2(1atm), DCM, methanol, RT, 30min, 22% -89%.
The substituents R for compounds 45a-j and compounds 46a-j are shown in Table 1 below:
[ TABLE 1 ]
(reaction formula 7) preparation of 5-Hydroxypyridine derivative (5-Hydroxypyridine Derivatives) (Compound 47)
The Boc deprotection reagents and conditions of scheme 7: 20% TFA, DCM, RT, 1h, 67%.
(reaction formula 8) preparation of 5-Hydroxypyridine derivative (5-Hydroxypyridine Derivatives) (Compound 48)
Reagents and conditions for the diels-Alder (diels-Alder) reaction of equation 8: ethyl crotonate, pure, 5h, 42%.
Since the compounds represented by the general formula I of the present invention exhibit strong antagonistic actions against P2X1 receptors and P2X3 receptors, the above compounds can be effectively used as agents capable of preventing or treating diseases caused by the activities of P2X1 receptors and P2X3 receptors, such as chronic inflammatory diseases such as degenerative arthritis, rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, cystitis, and the like; neuropathic pain diseases such as neuropathic pain, allodynia, diabetic neuropathy, spontaneous pain, irritative pain, phantom limb pain, or complex regional pain syndrome; or diseases associated with platelet aggregation such as arteriosclerosis, apoplexy, thrombosis, embolism, myocardial infarction, atherosclerosis or peripheral blood circulation disorder.
The present invention provides a pharmaceutical composition for preventing or treating chronic inflammatory diseases, neuropathic pain diseases or diseases associated with platelet aggregation caused by the activities of P2X1 receptor and P2X3 receptor, which contains the compound of the above general formula I as an active ingredient.
Chronic inflammatory diseases caused by the activity of the P2X1 receptor and the P2X3 receptor as defined herein include, for example, degenerative arthritis, rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, or cystitis, etc.; neuropathic pain disorders resulting from the activity of P2X1 receptors and P2X3 receptors include, for example, neuropathic pain, allodynia, diabetic neuropathy, idiopathic pain, irritative pain, phantom limb pain or complex regional pain syndrome, etc.; diseases associated with platelet aggregation caused by the activity of P2X1 receptor and P2X3 receptor include, for example, arteriosclerosis, stroke, thrombosis, embolism, myocardial infarction, atherosclerosis or peripheral blood circulation disorder, etc.
Therefore, the invention provides an antagonist for a P2X1 receptor and a P2X3 receptor, wherein the pyridine carboxylic acid compound with the structure of the general formula I is used as an effective component.
The present inventors confirmed that the novel 5-hydroxypyridine compound shows strong antagonistic activity against P2X1 receptor and P2X3 receptor in frog eggs (xenopus laevis oocytes) expressed as cloned mouse P2X1 receptor and human P2X3 receptor using the two-electrode voltage clamp method (TEVC), and confirmed that the compound of the present invention has a high development possibility as a drug for treating and preventing chronic inflammatory diseases, neuropathic pain diseases or platelet aggregation-related diseases caused by P2X1 receptor and P2X3 receptor activities by analyzing accumulation of ethidium bromide in human HEK293 cells expressed as human P2X7 receptor, and completed the present invention.
In another aspect, the present invention relates to a pharmaceutical composition comprising pyridine carboxylic acids for preventing and treating diseases caused by activity of P2X1 receptor and P2X3 receptor and a method for treating the above diseases by using pyridine carboxylic acids.
In the present invention, the term "prevention" refers to all actions aimed at inhibiting the onset or delaying the onset of all diseases associated with the activity of P2X1 receptor and P2X3 receptor by administering a composition comprising 5-hydroxypyridines. The term "treatment" refers to all behaviors intended to ameliorate or beneficially modify all diseases associated with P2X1 receptor and P2X3 receptor activity by administering a pharmaceutical composition.
The compositions and methods of treatment comprising 5 hydroxypyridines of the present invention may be used not only in humans, but also in mammals such as cattle, horses, sheep, pigs, goats, camels, antelopes, dogs, cats that may develop all diseases associated with P2X1 and P2X3 receptor activity.
In addition, the present invention provides a pharmaceutical composition for preventing and treating all diseases related to the activities of P2X1 receptor and P2X3 receptor, comprising the 5-hydroxypyridine compound having the structure of the above general formula (I) as an active ingredient.
Compositions comprising the compounds of the present invention may also include suitable carriers, excipients, or diluents according to conventional methods in the art.
Carriers, excipients and diluents that may be included in the compositions of the invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.
The composition containing the compound of the present invention is used in the form of oral preparations such as powder, granule, tablet, capsule, suspension, emulsion, syrup, and aerosol, external preparations, suppositories, or sterile injections according to a conventional method in the art.
In detail, when formulating a formulation, the composition of the present invention can be prepared by using diluents or excipients such as fillers, binders, wetting agents, disintegrants, surfactants, etc., which are generally used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and may be prepared by mixing at least one excipient of the above-mentioned compounds, for example, starch, calcium carbonate (calcium carbonate), sucrose (sucrose), lactose, gelatin (lactose), and the like. Also, magnesium stearate and talc may be used in addition to simple excipients. Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups and the like. In addition to water and liquid paraffin, which are generally used as simple diluents, various excipients, for example, wetting agents, sweetening agents, flavoring agents, preservatives and the like, may be included. Formulations for non-oral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilizates and suppositories. Propylene glycol (propylene glycol), polyethylene glycol, vegetable oils (e.g., olive oil), injectable esters (e.g., ethyl oleate) can be used as the non-aqueous solution and suspension. As the base of the suppository, semisynthetic cocoyl ester (witepsol), polyethylene glycol (macrogol), tween 61, cacao butter, laurate, glycerogelatin, and the like can be used.
The preferred dosage of the compound of the present invention varies depending on the condition and body weight of the patient, the degree of disease, the pharmaceutical form, the administration route and time, but can be appropriately selected by those skilled in the art. However, in order to obtain preferable effects, the daily dose of the compound of the present invention may be 0.0001 to 100mg/kg, preferably 0.001 to 100mg/kg, and the dose may be administered once to several times per day. The compounds of the invention may be present in the composition in an amount of from 0.0001% to 10% by weight, preferably from 0.001% to 1% by weight, relative to the total weight of the composition.
In addition, pharmaceutical dosage forms of the compounds of the present invention may be employed in the form of pharmaceutically acceptable salts, either alone or in combination with other pharmaceutically active compounds and in appropriate combinations.
The pharmaceutical composition of the present invention is administered to mammals such as rats, mice, livestock and humans through various routes. All modes of administration are contemplated, for example, administration may be by oral, rectal or intravenous, intramuscular, subcutaneous, endometrial or intracerebroventricular (intracerebrave) injection.
Effects of the invention
The invention provides a novel 5-hydroxypyridine compound serving as a P2X1 and P2X3 receptor antagonist and a pharmaceutical composition containing the compound. The compounds according to the present invention strongly antagonize P2X1 and P2X3 receptors, and thus can be effectively used as drugs for the treatment and prevention of diseases caused by the activity of P2X1 and P2X3 receptors, i.e., chronic inflammatory diseases or neuropathic pain diseases.
Drawings
Fig. 1a to 1f are diagrams showing a P2X receptor antagonist developed in the prior art and a P2X receptor antagonist reported by the present inventors.
FIGS. 2a to 2c are graphs showing antagonistic activity against P2X3 and P2X2/3 receptors of Compound 29 of the present invention (2 a: hP2X3R, 2 b: rP2X 3/3).
Fig. 3a and 3b are graphs showing that the anti-hyperalgesic effect of SNL rats was shown by intradurally administering pregabalin (pregabalin), AF353 and compound 29. The withdrawal threshold (within the range of one hundred percent) or the percentage of the maximum possible effect (maximum possible effect) (% MPE at 1. mu.g) represents the mean. + -. SEM of 5 to 6 rats per experimental group. P < 0.05 compared to vehicle (control).
Fig. 4a and 4b are graphs showing the anti-hyperalgesic effect in SNL rats by intravenous administration of compound 28. The withdrawal threshold or percentage of maximum possible effect represents the mean ± SEM of 5-6 rats per experimental group. P <0.01, P <0.001, compared to vehicle (control).
Fig. 5a and 5b are graphs showing the anti-hyperalgesic effect of CIPN rats by intravenous administration of compound 28. The withdrawal threshold or percentage of maximum possible effect represents the mean ± SEM of 5-6 rats per experimental group. P <0.001 compared to vehicle (control).
Detailed Description
Hereinafter, the present invention will be described in more detail by examples. It will be apparent to those skilled in the art that these examples are intended to illustrate the present invention more specifically, and the scope of the present invention is not limited to these examples in light of the gist of the present invention.
Reference example 1, preparation of experiment, etc
Abbreviations
The abbreviations used in this experiment are defined as follows:
ACN, acetonitrile;
ANOVA, analysis of variance;
ATP, 5' -adenosine triphosphate;
BBB, blood brain barrier;
BzATP, 2 '(3') -O- (4-benzoylbenzoyl) -ATP;
CIPN, chemotherapy-induced peripheral neuropathy;
CYP, cytochrome P450;
DMSO, dimethyl sulfoxide;
FBS, fetal bovine serum;
i.p., intraperitoneal administration;
i.v., intravenous injection;
MPE, maximum possible effect;
NeP, neuropathic pain;
PBS, phosphate buffered saline;
PEG, polyethylene glycol;
p2XR, P2X receptor;
SAR, structure-activity relationship;
SNL, spinal nerve ligation;
TEVC, dual electrode voltage clamp;
TM, transmembrane
Chemical analysis
1H and13c Nuclear Magnetic Resonance (NMR) spectra were obtained from a 300MHz Jeol JNM-LA 300WB spectrometer and a 400MHz Jeol JNM-ECX 400P spectrometer and from CDCl3，DMSO-d6Or CD3OD. Chemical shifts are expressed in δ units and ppm units relative to Tetramethylsilane (TMS) as an internal standard. The data are reported as follows: chemical shifts (Chemical shifts), integration (integration), multiplicities (s, singlet; d, doublet; t, triplet; m, multiplet; br, broad). Coupling constants J are reported in Hz units. Mass spectrometry was performed on MALDI-TOF and ESI (electrospray ionization) equipment. In addition, high resolution mass spectra (m/z) of selected compounds were separated by ESI (electrospray ionization) at Korea (Changxingshire)And (6) analyzing.
The purity of all final compounds was determined by HPLC (purity at least 95% unless otherwise indicated). The determination of purity was performed by using a Shimadzu Shim-pack C18 analytical column (250 mm. times.4.6 mm, 5 μm, 100A) on a Shimadzu SCL-10A VP HPLC system with two solvent systems. By passing through a 0.1% formic acid solution (H)2O):CH3The solvent system was run at a flow rate of 1mL/min for 30 minutes at a CN-90: 10 to 1:100 ratio. The peak (peak) was detected by UV absorption using a diode array detector.
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