Pyrrolo [2,3-B ] pyridine derivatives as inhibitors of influenza virus replication
阅读说明：本技术 作为流感病毒复制的抑制剂的吡咯并[2,3-b]吡啶衍生物 (Pyrrolo [2,3-B ] pyridine derivatives as inhibitors of influenza virus replication ) 是由 伊里娜·C·雅各布森 塞姆·斯科·李 迈克尔·大卫·费思 于 2019-07-26 设计创作，主要内容包括：本文提供式A、B或C的化合物,其可抑制流感病毒的复制,减少流感病毒的量和/或治疗流感。(Provided herein are compounds of formula A, B or C that can inhibit replication of influenza virus, reduce the amount of influenza virus, and/or treat influenza.)
1. A compound having formula A, B or C:
wherein:
Z1is-R, halogen, cyano, -OR, -CO2R*、-NO2or-CON (R)2；
Z2is-R, -OR, -CO2R*、-NR*2or-CON (R)2；
Z3is-H, hydroxy, halogen, -NH2；-NH(C1-C6Alkyl groups); -N (C)1-C6Alkyl radical)2、C1-C6Alkoxy or C1-C6Alkyl optionally substituted with one or more substituents independently selected from the group consisting of: halogen, cyano, hydroxy and C1-C6An alkoxy group;
x is-C (R)5)2-、-O-、-N(R5)-、-NR5-SO2-、C6-C10Aryl or 5-to 10-membered heteroaryl comprising 1 to 3 ring heteroatoms selected from O, N and S,
wherein when X is-N (R)5) When is, R5The moiety together with the nitrogen to which it is attached optionally forms a 5 or 6 membered heterocyclic ring, in which case L is attached to a ring atom (e.g. a ring carbon or nitrogen atom), preferably to a carbon or nitrogen ring atom that is beta to the nitrogen, and the 5 or 6 membered heterocyclic ring may optionally include one or two oxo substituents,
R1is-H or C1-C6An alkyl group;
R2is-H; -F; -NH2；-NH(C1-C6Alkyl groups); -N (C)1-C6Alkyl radical)2(ii) a -C ═ N-OH; cyclopropyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, -OCH3and-CH3(ii) a Or C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy and C 1-C6An alkoxy group; and
R3is-H, halogen, hydroxy, C1-C6Alkoxy, -NH2、-NH(C1-C6Alkyl), -N (C)1-C6Alkyl radical)2-cyano or C1-C6Alkyl radical, wherein C1-C6Alkyl is optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C6Alkyl), -N (C)1-C6Alkyl radical)2、-OCO-C1-C6Alkyl, -CO-C1-C6Alkyl, -CO2H、-CO2C1-C6Alkyl and C1-C6An alkoxy group;
each R4Independently H, C1-C6Alkyl radical, C3-C8Cycloalkyl radicals、C2-C8Alkoxy group, (C)1-C14Alkoxy group)r-(C1-C14Alkyl radical)s、C2-8alkoxy-C6-C10Aryl radical, C1-C6alkyl-C6-C10Aryl, 5-to 10-membered heteroaryl or C1-C6An alkyl-5 to 10 membered heteroaryl, wherein the heteroaryl comprises 1 to 3 ring heteroatoms selected from O, N and S;
each R5Independently H, C1-C6Alkyl radical, C3-C8Cycloalkyl radical, C2-C8Alkoxy radical, C2-C8alkoxy-C6-C10Aryl radical, CO2H、CO2-C1-C6Alkyl, CONH2、CONH-C1-C6Alkyl, CON (C)1-C6Alkyl radical)2、C1-C6alkyl-CO2H、C1-C6alkyl-CO2-C1-C6Alkyl radical, C1-C6alkyl-CONH2、C1-C6alkyl-CONH (C)1-C6Alkyl group), C1-C6alkyl-CON (C)1-C6Alkyl radical)2、C1-C6alkyl-C6-C10Aryl, 5-to 10-membered heteroaryl or C1-C6An alkyl-5 to 10 membered heteroaryl, wherein the heteroaryl comprises 1 to 3 ring heteroatoms selected from O, N and S;
R9independently-H, halogen, cyano, hydroxy, -NH2Carboxyl group, C1-C6Alkyl radical, C1-C6Haloalkyl, C 1-C6Cyanoalkyl, C2-C6alkoxy-C1-C6Alkyl radical, C1-C6Aminoalkyl radical, C1-C6Hydroxyalkyl, CO-C1-C6Alkyl or C1-C6An alkoxy group;
each R is independently: i) -H; ii) C1-C6Alkyl, optionally via one or more (e.g. 1, 2 or 3)One) is independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2Carboxyl group, C3-C8Cycloalkyl, 5-to 6-membered heterocycloalkyl, phenyl, 5-to 6-membered heteroaryl, C1-C6Alkoxy and-CO-C1-C6An alkyl group; wherein C is1-C6Alkoxy and-CO-C1-C6Each of said alkyl groups in an alkyl group is optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO-C1-C4Alkyl, -CO-C1-C4Alkyl, -CO2H、-CO2-C1-C4Alkyl and C1-C4Alkoxy, wherein the heterocycloalkyl or heteroaryl includes 1 to 3 ring heteroatoms selected from O, N and S, and wherein each of the cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl is independently and optionally substituted with one or more (e.g., 1, 2, or 3) instances of J; or iii) C3-C8Cycloalkyl or 4 to 8 membered heterocycloalkyl comprising 1 to 3 ring heteroatoms selected from O, N and S, each independently and optionally substituted with one or more (e.g., 1, 2 or 3) instances of J; and
Each J is independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2Carboxyl group, amide group, C1-C6Alkyl, -C1-C6Alkoxy and-CO-C1-C6Alkyl, wherein each of said alkyl groups is optionally substituted with one or more (e.g., 1, 2, or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO-C1-C4Alkyl, -CO2H、-CO2C1-C4Alkyl and C1-C4Alkoxy radical；
L is (C)1-C14Alkoxy group)q-Rz、C1-C14Alkyl radical- (C)1-C14Alkoxy group)q-Rz、(C1-C14Alkoxy group)q-Y、C1-C14alkyl-C (O) NR5-C1-C14alkyl-Rz、C1-C14alkyl-NR5C(O)-C1-C14alkyl-Rz、C1-C14alkyl-OC (O) NH-C1-C14alkyl-Rz、-(C1-C14Alkyl radical)q-(C1-C14Alkoxy group)q-Y-(C1-C14Alkyl radical)q-Rz、(C1-C14Alkoxy group)q-Y-(C1-C14Alkoxy group)q-(C1-C14Alkyl radical)q-Rz、(C1-C14Alkyl radical)q-Y-C1-C14Alkyl radical- (C)1-C14Alkoxy group)q-Rz；
Each RzIndependently selected from H, halogen, cyano, hydroxy, C1-C6Alkyl radical, C1-C6Haloalkyl, -OR5、(C1-C6Alkoxy group)r-O-C1-C6Haloalkyl, C1-6alkyl-NHR5、-N(R5)2、-NH(R5)-CO2H、C1-C6alkoxy-OCHO, -CO2R5、C1-C6alkyl-CO2R5、-CONHR5、-O-Y、-CONH(R5)-Y、CO-Y、C1-C14alkyl-Y, C1-C14alkoxy-Y and Y;
each Y is independently selected from C6-C10Aryl radical, C3-C12Cycloalkyl, 5-to 10-membered heteroaryl, or 3-to 12-membered heterocyclyl, wherein said heteroaryl or heterocyclyl includes 1 to 3 ring heteroatoms selected from O, N and S, wherein Y is optionally substituted with one to three substituents selected fromAnd (3) substituent: halogen, cyano, hydroxy, C 1-C6Alkyl radical, C1-C6Alkoxy, -CO2H、-CO2C1-C4Alkyl and-O-C6-C10An aryl group;
m is 0, 1, 2 or 3;
q is 0, 1, 2 or 3 and if X is not-C (R)5)2-or 5 to 10 membered heteroaryl, then q is 1, 2 or 3; and
each of r and s is 0, 1, 2 or 3; and r + s is 1 to 6,
or a pharmaceutically acceptable salt thereof.
2. The compound or salt of claim 1 having the structure of any one of formulas 1 to 13:
wherein:
x is-C (R)5)2-、-N(R5)-、-NR5-SO2-or-O-;
each Q is independently N, CH or C, wherein indicates the point of attachment of the L group and only one Q is C; and
n is independently 1, 2, 3 or 4.
3. The compound or salt of claim 1 or 2, wherein L is one of the following formulae:
-CH2CH2OCH2CH2OCH2CH2-T, wherein T is selected from the group consisting of: OH, OR5、NHR5、CO2R5、CO2NHR5、C1-6alkyl-CO2R5、C1-6alkyl-NHR5、C6-C10Aryl, 5-to 10-membered heteroaryl, C1-8Alkyl and C3-8A cycloalkyl group;
-C1-8alkoxy-C1-8alkoxy-U;
-C1-8alkoxy-C1-8alkoxy-C1-8An alkoxy group-U, which is a cyclic alkoxy group,
-C1-8alkoxy-aryl/heteroaryl-C1-8alkyl-C1-8Alkoxy radical
-C6-13alkyl-U;
-C1-8alkyl-C1-8alkoxy-U;
-C1-8alkyl-C1-8alkoxy-C1-8alkoxy-U;
-C1-8alkyl-C1-8alkoxy-C1-8alkoxy-C1-8alkoxy-U;
-C1-8alkyl-aryl/heteroaryl-C1-8alkoxy-C1-8alkoxy-U;
-C1-8alkyl-aryl/heteroaryl-C 1-8alkyl-C1-8alkoxy-C1-8alkoxy-U;
-C1-8alkyl-aryl/heteroaryl-C1-8alkyl-C1-8alkoxy-C1-8alkyl-U;
-C1-8alkyl-aryl/heteroaryl-C1-8alkyl-C1-8alkoxy-U;
-C1-8alkyl-C1-8alkoxy-aryl/heteroaryl-C1-8Alkoxy-aryl;
-C1-8alkyl-C1-8alkoxy-aryl/heteroaryl-C1-8alkyl-U;
-C1-8alkyl-C1-8alkoxy-aryl/heteroaryl-C1-8alkyl-alkoxy-U;
-C1-8alkyl- (optionally substituted) phenoxy;
-C1-8alkyl-C1-8Alkoxy- (optionally substituted) phenoxy;
-C1-8alkyl-N (R)5)-C(O)-C1-8alkyl-C1-8alkoxy-U;
-C1-8alkyl-O-C (O) N (R)5)-C1-8alkyl-U;
-C1-8alkyl-aryl/heteroaryl-C1-8alkoxy-U;
-aryl/heteroaryl-U;
-aryl/heteroaryl-C1-8alkoxy-U;
-aryl/heteroaryl-C1-8alkoxy-C1-8alkyl-U;
-aryl/heteroaryl-C1-8alkoxy-C1-8alkoxy-U;
-aryl/heteroaryl-C1-8alkyl-C1-8alkoxy-C1-8alkoxy-U;
-aryl/heteroaryl-C1-8alkoxy-C1-8alkoxy-U; u is selected from the group consisting of: H. c1-8Alkoxy radical, C6-C10Aryloxy group, OR5、NHR5、CO2R5、C(O)NHR5、C1-6alkyl-CO2R5、C1-6alkyl-NHR5、C6-C10Aryl radical, C6-C10aryl-C1-6Alkyl radical, C1-6alkyl-C6-C10Aryl, 3-to 12-membered heterocycloalkyl, 5-to 10-membered heteroaryl, C1-8Alkyl radical, C1-6Haloalkyl and C3-8Cycloalkyl, wherein the heterocycloalkyl and heteroaryl have 1 to 3 ring heteroatoms selected from N, O and S,
Wherein the alkyl, alkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, and cycloalkyl moieties may be optionally substituted with one to three (e.g., 1, 2, or 3) substituents selected from the group consisting of: hydroxy, C1-6Hydroxyalkyl radical, C1-6Alkoxy radical, C2-8Alkoxyalkyl group, C2-8Alkyl alkoxy radical, C1-8Alkoxycarbonyl group, C1-8Alkyl, arylalkoxy CO-, C2-6Alkenyl radical, C2-6Alkynyl, carboxyl, halogen, C1-6A haloalkyl group,N3Cyano, N (R')2、SR'、-C(O)NHR'、OCOR'、OC(O)NHR'、N(CO)R'、N(CO)OR'、N(CO)COR'、SCOR'、S(O)2NR'2、S(O)2R ', wherein each R' is independently H, C1-6Alkyl radical, C1-6Haloalkyl, C1-6Alkoxy radical, C2-6Alkenyl radical, C2-6Alkynyl, C3-6Cycloalkyl, heterocyclyl, aryl, aryloxy, heteroaryl, heteroaryloxy, alkylaryl, arylalkyl, heteroarylalkyl, or alkylheteroaryl, and said heteroaryl having 1 to 3 ring heteroatoms selected from N, O and S.
4. The compound or salt of claim 1 or 2, wherein L is selected from S1 to S57:
5. the compound or salt of any one of claims 1 to 4, wherein Z1And R3Each independently is F or Cl, and R1、R2、Z2And Z3Each is H.
6. The compound or salt of any one of claims 1 to 5, wherein Z1And R3One or each of which is F.
7. The compound or salt according to any one of claims 1 to 4, wherein:
R1is-H;
R2is-H;
R3is-H, -F, -Cl, C1-4Alkyl or C1-4A haloalkyl group;
Z1is-H, -F or-Cl;
Z2is-H;
Z3is-H; and is
R9Independently is-H or C1-4An alkyl group.
8. The compound or salt of any one of claims 1 to 7, having the structure:
wherein
Het/Ar is C6-C10Aryl or 5 to 10 membered heteroaryl having 1 to 3 ring heteroatoms selected from N, O and S; and
l is C1-C14alkyl-NR5C(O)-C1-C14alkyl-RzOr C1-C14alkyl-OC (O) NH-C1-C14alkyl-Rz(ii) a And
Rzis H, C1-C6Alkyl or C1-C6alkyl-CO2R5And said alkyl is optionally substituted with one or more (e.g. 1, 2 or 3) of: hydroxy, halogen, C1-C6Alkoxy radical, C1-C6Haloalkoxy, NH2、
-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO-C1-C4Alkyl, -CO2H and-CO2C1-C4An alkyl group.
9. A compound or salt according to claim 8, wherein Het/Ar is pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, furanyl, pyranyl, thienyl, benzimidazolyl, benzothienyl, benzofuranylbenzotriazolyl, benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, indolyl, isoindolyl, purinyl, phenyl, or naphthyl.
10. A compound or salt according to claim 8 or 9, wherein Het/Ar is pyridinyl or phenyl.
11. A compound or salt according to any one of claims 8 to 10, wherein Het/Ar is pyridinyl.
12. The compound or salt of any one of claims 8 to 11 wherein L is C1-C14alkyl-OC (O) NH-C1-C14alkyl-Rz。
13. A compound or salt of claim 12 wherein L is C1-C6alkyl-OC (O) NH-C1-C6alkyl-Rz。
14. The compound or salt of claim 1 selected from the group consisting of:
c1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13 and C14.
15. The compound or salt of claim 1 selected from the group consisting of:
a1, a2, A3, a4, a5, a7, A8, a9, a10, a11, a12, a13, a14, and a 15.
16. The compound or salt of claim 1 selected from the group consisting of: a23, a24, a27, a28, a29, a30, a31, a33, a34, a35, a36, a37, a38, a39, a40, a41, a42, a43, a44, a45, a46, a47, a48, a49, a50, a51, a52, a53, a54, a55, a56, a57, a58, a59, a60, a61, a62, a63, a64, a65, a66, a67, a 36160, a153, a140, a124, a119, a124, a119, a124, a119, a126, a119, a126, a124, a129, a112, a124, a119, a126, a124, a126, a129, a112, a124, a112, a126, a112, a119, a124, a119, a124, a126, a124, a112, a115, a124 a115, a.
17. The compound or salt of claim 1 selected from the group consisting of: a164, a165, a166, a167, a168, a169, a170, a171, a172, a173, a174, a175, a176, a177, a178, a179, a180, a181, a182, a183, a184, a185, a186, a187, a188, a189, a190, a191, a192, a193, a194, a195, a196, a198, a199, a200, a201, a202, a203, a204, a205, a206, a207, a208, a209, a210, a211, a212, a213, a214, a215, a216, a219, a220, a221, a222, a223, a224, a225, a243, a227, a228, a229, a248, a230, a231, a232, a233, a234, a235, a236, a237, a236, a239, a240, a245, a246, and a 247.
18. The compound or salt of claim 1 selected from the group consisting of: a263, a264, a265, a266, a267, a268, a269, a270, a271, a272, a273, a274, a275, a276, a277, a278, a279, a280, a281, a282, a283, a284, a285, a286, a287, a288, a289, and a 290.
19. The compound or salt of claim 1 selected from the group consisting of: b1, B2, B3, B4, B5, B6 and B7.
20. The compound or salt of claim 1, having the formula:
wherein R' is-H or C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, amino, carboxy, C1-C6Alkoxy radical, C1-C6Haloalkoxy, C1-C6Aminoalkoxy group, C1-C6Cyanoalkoxy group, C1-C6Hydroxyalkoxy and C2-C6An alkoxyalkoxy group.
21. The compound or salt of claim 1, having the formula:
wherein R' is-H or C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, amino, CO2H、C1-C6Alkoxy radical, C1-C6Haloalkoxy, C1-C6Aminoalkoxy group, C1-C6Cyanoalkoxy group, C1-C6Hydroxyalkoxy and C2-C6An alkoxyalkoxy group.
22. The compound of claim 20 or 21, wherein R "is C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, hydroxy, amino, carboxy, C1-C6Alkoxy radical, C1-C6Haloalkoxy, C1-C6Aminoalkoxy group, C1-C6Hydroxyalkoxy and C2-C6An alkoxyalkoxy group.
23. A compound as listed in table a or a pharmaceutically acceptable salt thereof.
24. A pharmaceutical composition comprising a compound or salt according to any one of claims 1 to 23 and a pharmaceutically acceptable carrier, adjuvant or vehicle.
25. A method of treating or preventing influenza in a patient comprising administering to the patient a safe and effective amount of a compound or salt according to any one of claims 1 to 23.
26. A method of reducing the amount of influenza virus in a biological sample or patient comprising administering to the biological sample or patient a safe and effective amount of a compound or salt of any one of claims 1-23.
27. The method of claim 25 or 26, further comprising administering an influenza vaccine to the patient before, after, or simultaneously with the compound.
28. The method of any one of claims 25-27, wherein the compound is administered via a liposome or a nonionic surfactant vesicle (niosome).
29. The method of claim 28, wherein the liposome is a smaller unilamellar vesicle.
30. The method of claim 28 or 29, wherein the liposome is administered via intravenous injection or via inhalation.
31. The method of any one of claims 25 to 30, wherein the compound is administered via inhalation.
32. The method of claim 31, wherein the compound is provided in the form of porous particles sized to deliver the particles to the alveolar region of the lung.
33. Use of a compound or salt according to any one of claims 1 to 23 for the preparation of a medicament for treating or preventing influenza infection in a human.
34. Use of a compound according to any one of claims 1 to 23 for the preparation of a medicament for reducing the amount of influenza virus in a human.
35. The use of claim 33 or 34, wherein the medicament further comprises one or more additional active agents selected from the group consisting of: neuraminidase inhibitors, ion channel inhibitors and polymerase inhibitors.
36. The use of any one of claims 33-35, wherein the medicament is in the form of a liposome formulation.
37. The use of claim 36, wherein the liposome formulation comprises smaller unilamellar vesicles.
38. The use of any one of claims 33-37, wherein the medicament is formulated for intravenous administration.
39. The use of any one of claims 33-37, wherein the medicament is formulated for pulmonary or intranasal administration.
40. The use of claim 39, wherein the agent comprises porous particles sized for pulmonary administration to an alveolar region of a lung.
41. A composition comprising a compound or salt according to any one of claims 1 to 23 and a therapeutically effective amount of one or more additional active agents selected from the group consisting of: a neuraminidase inhibitor, an ion channel inhibitor and a polymerase inhibitor, wherein the combined amounts have a therapeutic effect on treating an influenza infection in a subject.
Technical Field
The present disclosure relates generally to inhibitors of influenza virus replication, and methods of treating or preventing influenza infection by administering the inhibitors to a patient in need of treatment.
Background
Influenza spreads worldwide in a seasonal fashion, resulting in hundreds of thousands of deaths per year, with millions in pandemic years. For example, three influenza pandemics occurred in the 20 th century, and killed tens of millions of people, with each pandemic being caused by the emergence of novel strains of human viruses. Typically, these novel strains arise as a result of transmission of existing influenza viruses from other animal species to humans.
Influenza is transmitted from person to person primarily via larger virus-carrying droplets produced when an infected person coughs or sneezes; these larger droplets may then settle on the mucosal surfaces of the upper respiratory tract of susceptible individuals in the vicinity of (e.g., within about 6 feet of) an infected person. Transmission can also occur via direct contact or indirect contact with respiratory secretions (such as touching a surface contaminated with influenza virus and then touching the eyes, nose or mouth). Adults may transmit influenza to others within 1 day before symptoms appear to about 5 days after symptoms begin. Young children and people with weak immune systems may remain contagious 10 or more days after the onset of symptoms.
Influenza viruses are RNA viruses of the orthomyxoviridae family, which includes five genera: influenza A virus, influenza B virus, influenza C virus, infectious salmon anemia virus (Isavirus) and Togao virus (Thogoto virus).
Influenza a viruses can cause seasonal influenza and pandemic influenza epidemics. It has one species, influenza a virus, and wild waterfowl is the natural host for many influenza a viruses. Occasionally, the virus spreads to other species and can then cause destructive outbreaks in poultry or produce human influenza pandemics. Type a viruses are the most virulent human pathogens of the three influenza types and cause the most serious diseases. Influenza a viruses can be subdivided into different serotypes based on antibody responses to these viruses. Among the serotypes identified in humans, ranked by the number of known human pandemic deaths, are: H1N1 (which caused spanish influenza in 1918), H2N2 (which caused asian influenza in 1957), H3N2 (which caused hong kong influenza in 1968), H5N1 (pandemic threat in influenza season between 2007 and 2008), H7N7 (which is a potential pandemic threat), H1N2 (endemic disease present in humans and pigs), H9N2, H7N2, H7N3 and H10N 7.
Influenza B virus can cause seasonal influenza and has one species, influenza B virus. Influenza B almost completely infects humans and is less prevalent than influenza a. The only other animal known to be susceptible to influenza B infection is seal. This type of influenza mutates at a rate 2 to 3 times slower than type a and is therefore less genetically diverse, having only one influenza B serotype. Due to this lack of antigenic diversity, a certain degree of immunity to influenza B is usually obtained early. However, influenza B is sufficiently mutated that sustained immunity is not possible. This reduced rate of antigenic change, combined with its restricted host range (inhibition of cross species antigen transfer), ensures that a pandemic of influenza B does not occur.
Influenza C virus has one species, influenza C virus, which infects humans and pigs and can cause severe disease and local epidemics. However, influenza C is less prevalent than other types and appears to often cause mild disease in children.
The structures of influenza viruses of each serotype and genus are very similar. The influenza genome consists of eight single-stranded RNAs packaged into rod-like structures of different sizes, called ribonucleoprotein complexes (RNPs). Each RNP contains a unique viral RNA, multiple copies of backbone nucleoproteins, and a heterologous trimeric viral polymerase consisting of PA, PB1, and PB2 subunits, which catalyzes transcription and replication of the viral genome. Recent biochemical and structural studies of the influenza polymerase complex provide insight into the mechanistic understanding of cap-robbing (cap-snatching) and RNA synthesis by influenza polymerase. Briefly, the PB2 end cap binding domain first binds to its 5' end cap to isolate the host precursor mRNA. PA, endonuclease subunit, followed by decomposition of 10-13 nucleotides of captured pre-mRNA downstream of the end cap. The PB2 subunit was then spun about 700 to direct the capping primer into the PB1 polymerase active site. The PB1 subunit interacts directly with the PB2 and PA subunits. These subunits contain highly conserved domains in different influenza strains and are attracted as attractive anti-influenza drug targets. In addition to the polymerase complex, the influenza genome encodes its own Neuraminidase (NA), Hemagglutinin (HA), Nucleoprotein (NP), matrix proteins M1 and M2 and nonstructural proteins NS1 and NS 2. NA is the target of the antiviral drugs oseltamivir (Tamiflu) and zanamivir (zanamivir) (renenza). These drugs are sialic acid analogs that inhibit the enzymatic activity of NA, thereby slowing the release of progeny virus from infected cells.
Influenza incurs direct costs due to lost productivity and related medical treatments, and indirect costs due to preventive measures. In the united states, influenza costs a total of over 100 billion dollars per year, while it is estimated that future pandemics can cause direct and indirect costs in the billions of dollars. The cost of control is also high. Governments worldwide have spent billions of dollars in preparing and planning for a potential H5N1 avian influenza pandemic, with costs associated with purchasing medications and vaccines and conducting disaster maneuvers and developing strategies for improved border control.
Current treatment options for influenza include vaccination and chemotherapy or chemoprevention using antiviral drugs. Vaccination against influenza with influenza vaccines is generally recommended for high risk groups such as children and the elderly, or people with asthma, diabetes or heart disease. However, it is possible that influenza still develops after vaccination. The vaccine is re-formulated for several specific influenza virus strains each season, but may not contain all the strains that effectively infect people worldwide during that season. Manufacturers spend about six months making up and producing millions of doses needed to handle seasonal behavior; occasionally, new or overlooked virus strains become prominent during that period and infect already vaccinated people (e.g. H3N2 fujian influenza in the 2003 to 2004 flu season). It may also happen to be infected just before vaccination and with the particular viral strain that the vaccine should prevent, since it takes about two weeks for the vaccine to be effective.
Furthermore, the effectiveness of these influenza vaccines is variable. Because of the high mutation rate of viruses, certain influenza vaccines typically provide protection for no more than a few years. A vaccine formulated for one year may not be effective in the next year because influenza viruses change rapidly over time and different virus strains become dominant.
Due to the absence of RNA-correcting enzymes, the RNA-dependent RNA polymerase of influenza vRNA generates a nucleotide insertion error approximately every ten thousand nucleotides (which is approximately the length of influenza vRNA). Thus, almost every newly manufactured influenza virus has a mutant antigen drift. The separation of the genome into eight separate segments of vRNA allows for mixing or reassortment of vRNA when more than one strain infects a single cell. The resulting rapid changes in viral genetics produce antigen transfer and allow the virus to infect novel host species and rapidly overcome protective immunity.
Antiviral drugs may also be used to treat influenza, where NA inhibitors are particularly effective, but the virus may develop resistance to approved NA antiviral drugs. Furthermore, the emergence of multi-drug resistant pandemic influenza a virus has been well documented. Drug resistant pandemic influenza a becomes a significant public health threat. In addition to resistant influenza a viruses, NA inhibitors are approved for the treatment of early stage influenza infection (within 48 hours of onset of influenza symptoms).
Thus, there remains a need for drugs for treating influenza infections, such as drugs with an extended therapeutic window and/or reduced sensitivity to viral titers.
Disclosure of Invention
The present disclosure relates generally to methods of treating influenza, methods of inhibiting replication of influenza virus, methods of reducing the amount of influenza virus, compounds and compositions useful in such methods.
In one aspect, the present disclosure provides compounds of formulae A, B and C and pharmaceutically acceptable salts thereof:
wherein:
Z1is-R, halogen, cyano, -OR, -CO2R*、-NO2or-CON (R)2；
Z2is-R, -OR, -CO2R*、-NR*2or-CON (R)2；
Z3is-H, hydroxy, halogen, -NH2；-NH(C1-C6Alkyl groups); -N (C)1-C6Alkyl radical)2、C1-C6Alkoxy or C1-C6Alkyl optionally substituted with one or more substituents independently selected from the group consisting of: halogen, cyano, hydroxy and C1-C6An alkoxy group;
x is-C (R)5)2-、-O-、-N(R5)-、-NR5-SO2-、C6-C10Aryl or 5-to 10-membered heteroaryl comprising 1 to 3 ring heteroatoms selected from O, N and S,
wherein when X is-N (R)5) When is, R5The moiety together with the nitrogen to which it is attached optionally forms a 5 or 6 membered heterocyclic ring, in which case L is attached to a ring atom (e.g. a ring carbon or nitrogen atom), preferably to a carbon or nitrogen ring atom that is beta to the nitrogen, and the 5 or 6 membered heterocyclic ring may optionally include one or two pendant oxy substituents,
R1is-H or C1-C6An alkyl group;
R2is-H; -F; -NH2；-NH(C1-C6Alkyl groups); -N (C)1-C6Alkyl radical)2(ii) a -C ═ N-OH; cyclopropyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, -OCH3and-CH3(ii) a Or C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogenElement, cyano, hydroxy and C1-C6An alkoxy group; and
R3is-H, halogen, hydroxy, C1-C6Alkoxy, -NH2、-NH(C1-C6Alkyl), -N (C)1-C6Alkyl radical)2-cyano or C1-C6Alkyl radical, wherein C1-C6Alkyl is optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C6Alkyl), -N (C)1-C6Alkyl radical)2、-OCO-C1-C6Alkyl, -CO-C1-C6Alkyl, -CO2H、-CO2C1-C6Alkyl and C1-C6An alkoxy group;
each R4Independently H, C1-C6Alkyl radical, C3-C8Cycloalkyl radical, C2-C8Alkoxy group, (C)1-C14Alkoxy group)r-(C1-C14Alkyl radical)s、C2-8alkoxy-C6-C10Aryl radical, C1-C6alkyl-C6-C10Aryl, 5-to 10-membered heteroaryl or C1-C6An alkyl-5 to 10 membered heteroaryl, wherein the heteroaryl comprises 1 to 3 ring heteroatoms selected from O, N and S;
each R5Independently H, C1-C6Alkyl radical, C3-C8Cycloalkyl radical, C 2-C8Alkoxy radical, C2-C8alkoxy-C6-C10Aryl radical, CO2H、CO2-C1-C6Alkyl, CONH2、CONH-C1-C6Alkyl, CON (C)1-C6Alkyl radical)2、C1-C6alkyl-CO2H、C1-C6alkyl-CO2-C1-C6Alkyl radical, C1-C6alkyl-CONH2、C1-C6Alkyl radical-CONH(C1-C6Alkyl group), C1-C6alkyl-CON (C)1-C6Alkyl radical)2、C1-C6alkyl-C6-C10Aryl, 5-to 10-membered heteroaryl or C1-C6An alkyl-5 to 10 membered heteroaryl, wherein the heteroaryl comprises 1 to 3 ring heteroatoms selected from O, N and S;
R9independently-H, halogen, cyano, hydroxy, -NH2Carboxyl group, C1-C6Alkyl radical, C1-C6Haloalkyl, C1-C6Cyanoalkyl, C2-C6alkoxy-C1-C6Alkyl radical, C1-C6Aminoalkyl radical, C1-C6Hydroxyalkyl, CO-C1-C6Alkyl or C1-C6An alkoxy group;
each R is independently: i) -H; ii) C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2Carboxyl group, C3-C8Cycloalkyl, 5-to 6-membered heterocycloalkyl, phenyl, 5-to 6-membered heteroaryl, C1-C6Alkoxy and-CO-C1-C6An alkyl group; wherein C is1-C6Alkoxy and-CO-C1-C6Each of said alkyl groups in an alkyl group is optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO-C1-C4Alkyl, -CO-C 1-C4Alkyl, -CO2H、-CO2-C1-C4Alkyl and C1-C4Alkoxy, wherein the heterocycloalkyl or heteroaryl includes 1 to 3 ring heteroatoms selected from O, N and S, and wherein each of the cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl is independently and optionally via one or more (e.g., 1, 2, or 3) J heteroatomsSubstitution is carried out; or iii) C3-C8Cycloalkyl or 4 to 8 membered heterocycloalkyl comprising 1 to 3 ring heteroatoms selected from O, N and S, each independently and optionally substituted with one or more (e.g., 1, 2 or 3) instances of J; and
each J is independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2Carboxyl group, amide group, C1-C6Alkyl, -C1-C6Alkoxy and-CO-C1-C6Alkyl, wherein each of said alkyl groups is optionally substituted with one or more (e.g., 1, 2, or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO-C1-C4Alkyl, -CO2H、-CO2C1-C4Alkyl and C1-C4An alkoxy group;
l is (C)1-C14Alkoxy group)q-Rz、C1-C14Alkyl radical- (C)1-C14Alkoxy group)q-Rz、(C1-C14Alkoxy group)q-Y、C1-C14alkyl-C (O) NR5-C1-C14alkyl-Rz、C1-C14alkyl-NR5C(O)-C1-C14alkyl-Rz、C1-C14alkyl-OC (O) NH-C1-C14alkyl-Rz、-(C1-C14Alkyl radical)q-(C1-C14Alkoxy group)q-Y-(C1-C14Alkyl radical)q-Rz、(C1-C14Alkoxy group)q-Y-(C1-C14Alkoxy group) q-(C1-C14Alkyl radical)q-Rz、(C1-C14Alkyl radical)q-Y-C1-C14Alkyl radical- (C)1-C14Alkoxy group)q-Rz；
Each RzIndependently selected from H, halogen, cyano, hydroxy, C1-C6Alkyl radical, C1-C6Haloalkyl, -OR5、(C1-C6Alkoxy group)r-O-C1-C6Haloalkyl, C1-6alkyl-NHR5、-N(R5)2、-NH(R5)-CO2H、C1-C6alkoxy-OCHO, -CO2R5、C1-C6alkyl-CO2R5、-CONHR5、-O-Y、-CONH(R5)-Y、CO-Y、C1-C14alkyl-Y, C1-C14alkoxy-Y and Y;
each Y is independently selected from C6-C10Aryl radical, C3-C12A cycloalkyl, a 5-to 10-membered heteroaryl, or a 3-to 12-membered heterocyclyl, wherein the heteroaryl or heterocyclyl includes 1-3 ring heteroatoms selected from O, N and S, wherein Y is optionally substituted with one to three substituents selected from: halogen, cyano, hydroxy, C1-C6Alkyl radical, C1-C6Alkoxy, -CO2H、-CO2C1-C4Alkyl and-O-C6-C10An aryl group;
m is 0, 1, 2 or 3;
q is 0, 1, 2 or 3 and if X is not-C (R)5)2-or 5 to 10 membered heteroaryl, then q is 1, 2 or 3; and
each of r and s is 0, 1, 2 or 3; and r + s is 1 to 6.
In some cases, the compound is a compound of formulae 1 to 13, or a pharmaceutically acceptable salt thereof:
wherein:
x is-C (R)5)2-、-N(R5)-、-NR5-SO2-or-O-;
each Q is independently N, CH or C, wherein indicates the point of attachment of the L group and only one Q is C; and n is independently 1, 2, 3 or 4.
In some cases, the compound or a pharmaceutical salt thereof has the structure
Wherein
Het/Ar is C6-C10Aryl or 5 to 10 membered heteroaryl having 1 to 3 ring heteroatoms selected from N, O and S; and
L is C1-C14alkyl-NR5C(O)-C1-C14alkyl-RzOr C1-C14alkyl-OC (O) NH-C1-C14alkyl-Rz(ii) a And
Rzis H, C1-C6Alkyl or C1-C6alkyl-CO2R5And said alkyl is optionally substituted with one or more (e.g. 1, 2 or 3) of: hydroxy, halogen, C1-C6Alkoxy radical, C1-C6Haloalkoxy, NH2、
-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO-C1-C4Alkyl, -CO2H and-CO2C1-C4An alkyl group.
In some cases, the compound or a pharmaceutical salt thereof has formula (la)
Wherein R' is-H or C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxoRadical, amino group, carboxyl group, C1-C6Alkoxy radical, C1-C6Haloalkoxy, C1-C6Aminoalkoxy group, C1-C6Cyanoalkoxy group, C1-C6Hydroxyalkoxy and C2-C6An alkoxyalkoxy group.
In some cases, the compound or a pharmaceutical salt thereof has formula (la)
Wherein R' is-H or C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, amino, carboxy, C1-C6Alkoxy radical, C1-C6Haloalkoxy, C1-C6Aminoalkoxy group, C1-C6Cyanoalkoxy group, C1-C6Hydroxyalkoxy and C2-C6An alkoxyalkoxy group.
Further provided are methods of administering a safe and effective amount of a compound as disclosed herein, e.g., as represented by formula A, B or C or formulae 1 to 13, to a biological sample or patient.
Also provided herein are methods of reducing the amount of influenza virus in a biological sample or patient by administering to the biological sample or patient a safe and effective amount of a compound as disclosed herein, e.g., as represented by formula A, B or C or any of formulae 1 to 13.
Further provided are methods of treating or preventing influenza in a patient comprising administering to the patient a safe and effective amount of a compound as disclosed herein, e.g., as represented by formula A, B or C or formulae 1 to 13.
Also provided are pharmaceutical compositions comprising a compound as disclosed herein, e.g., as represented by formula A, B or C, or any one of formulas 1 to 13, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.
Also provided is the use of a compound described herein for inhibiting the replication of influenza virus in a biological sample or patient, for reducing the amount of influenza virus in a biological sample or patient, or for treating influenza in a patient.
Further provided herein is the use of a compound described herein for the manufacture of a medicament for treating influenza in a patient, for reducing the amount of influenza virus in a biological sample or patient, or for inhibiting replication of influenza virus in a biological sample or patient.
Detailed Description
Disclosed herein are compounds and the use of these compounds in inhibiting influenza virus. One aspect of the present disclosure relates generally to the use of a compound or pharmaceutically acceptable salt described herein, or a pharmaceutically acceptable composition comprising such a compound or pharmaceutically acceptable salt thereof, for inhibiting the replication of influenza virus in a biological sample or patient, for reducing the amount of influenza virus in a biological sample or patient (reducing viral titer), and for treating influenza in a patient.
Compounds of the present disclosure
The present disclosure provides compounds of formula A, B or C, or formulae 1 to 13, or pharmaceutically acceptable salts thereof:
or a pharmaceutically acceptable salt thereof, wherein:
Z1is-R, halogen, cyano, -OR, -CO2R*、-NO2or-CON (R)2；
Z2is-R, -OR, -CO2R*、-NR*2or-CON (R)2；
Z3is-H, hydroxy, halogen, -NH2；-NH(C1-C6Alkyl groups); -N (C)1-C6Alkyl radical)2、C1-C6Alkoxy or C1-C6Alkyl optionally substituted with one or more substituents independently selected from the group consisting of: halogen, cyano, hydroxy and C1-C6An alkoxy group;
x is-C (R)5)2-、-O-、-N(R5)-、-NR5-SO2-、C6-C10Aryl or 5-to 10-membered heteroaryl comprising 1 to 3 ring heteroatoms selected from O, N and S,
wherein when X is-N (R)5) When is, R5The moiety together with the nitrogen to which it is attached optionally forms a 5 or 6 membered heterocyclic ring, in which case L is attached to a ring atom (e.g. a ring carbon or nitrogen atom), preferably to a carbon or nitrogen ring atom that is beta to the nitrogen, and the 5 or 6 membered heterocyclic ring may optionally include one or two oxo substituents,
R1is-H or C1-C6An alkyl group;
R2is-H; -F; -NH2；-NH(C1-C6Alkyl groups); -N (C)1-C6Alkyl radical)2(ii) a -C ═ N-OH; cyclopropyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, -OCH3and-CH3(ii) a Or C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy and C1-C6An alkoxy group; and
R3is-H, halogen, hydroxy, C1-C6Alkoxy, -NH2、-NH(C1-C6Alkyl), -N (C)1-C6Alkyl radical)2-cyano or C1-C6Alkyl radical, wherein C1-C6Alkyl is optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C6Alkyl), -N (C)1-C6Alkyl radical)2、-OCO-C1-C6Alkyl, -CO-C1-C6Alkyl, -CO2H、-CO2C1-C6Alkyl and C1-C6An alkoxy group;
each R4Independently H, C1-C6Alkyl radical, C3-C8Cycloalkyl radical, C2-C8Alkoxy group, (C)1-C14Alkoxy group)r-(C1-C14Alkyl radical)s、C2-8alkoxy-C6-C10Aryl radical, C1-C6alkyl-C6-C10Aryl, 5-to 10-membered heteroaryl or C1-C6An alkyl-5 to 10 membered heteroaryl, wherein the heteroaryl comprises 1 to 3 ring heteroatoms selected from O, N and S;
each R5Independently H, C1-C6Alkyl radical, C3-C8Cycloalkyl radical, C 2-C8Alkoxy radical, C2-C8alkoxy-C6-C10Aryl radical, CO2H、CO2-C1-C6Alkyl, CONH2、CONH-C1-C6Alkyl, CON (C)1-C6Alkyl radical)2、C1-C6alkyl-CO2H、C1-C6alkyl-CO2-C1-C6Alkyl radical, C1-C6alkyl-CONH2、C1-C6alkyl-CONH (C)1-C6Alkyl group), C1-C6alkyl-CON (C)1-C6Alkyl radical)2、C1-C6alkyl-C6-C10Aryl, 5-to 10-membered heteroaryl or C1-C6An alkyl-5 to 10 membered heteroaryl, wherein the heteroaryl comprises 1 to 3 ring heteroatoms selected from O, N and S;
R9independently-H, halogen, cyano, hydroxy, -NH2Carboxyl group, C1-C6Alkyl radical, C1-C6Haloalkyl, C1-C6Cyanoalkyl, C2-C6alkoxy-C1-C6Alkyl radical, C1-C6Aminoalkyl radical, C1-C6Hydroxyalkyl, CO-C1-C6Alkyl or C1-C6An alkoxy group;
each R is independently: i) -H; ii) C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2Carboxyl group, C3-C8Cycloalkyl, 5-to 6-membered heterocycloalkyl, phenyl, 5-to 6-membered heteroaryl, C1-C6Alkoxy and-CO-C1-C6An alkyl group; wherein C is1-C6Alkoxy and-CO-C1-C6Each of said alkyl groups in an alkyl group is optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical) 2、-OCO-C1-C4Alkyl, -CO-C1-C4Alkyl, -CO2H、-CO2-C1-C4Alkyl and C1-C4Alkoxy, wherein the heterocycloalkyl or heteroaryl includes 1 to 3 ring heteroatoms selected from O, N and S, and wherein each of the cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl is independently and optionally substituted with one or more (e.g., 1, 2, or 3) instances of J; or iii) C3-C8Cycloalkyl or 4 to 8 membered heterocycloalkyl comprising 1 to 3 ring heteroatoms selected from O, N and S, each independently and optionally substituted with one or more (e.g., 1, 2 or 3) instances of J; and
each J is independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2Carboxyl group, amide group, C1-C6Alkyl, -C1-C6Alkoxy and-CO-C1-C6Alkyl, wherein each of said alkyl groups is optionally substituted with one or more (e.g., 1, 2, or 3) substituents independently selected from the group consisting of: halogenElements, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO-C1-C4Alkyl, -CO2H、-CO2C1-C4Alkyl and C1-C4An alkoxy group;
l is (C)1-C14Alkoxy group)q-Rz、C1-C14Alkyl radical- (C)1-C14Alkoxy group)q-Rz、(C1-C14Alkoxy group)q-Y、C1-C14alkyl-C (O) NR5-C1-C14alkyl-Rz、C1-C14alkyl-NR5C(O)-C1-C14alkyl-Rz、C1-C14alkyl-OC (O) NH-C1-C14alkyl-Rz、-(C1-C14Alkyl radical)q-(C1-C14Alkoxy group)q-Y-(C1-C14Alkyl radical) q-Rz、(C1-C14Alkoxy group)q-Y-(C1-C14Alkoxy group)q-(C1-C14Alkyl radical)q-Rz、(C1-C14Alkyl radical)q-Y-C1-C14Alkyl radical- (C)1-C14Alkoxy group)q-Rz；
Each RzIndependently selected from H, halogen, cyano, hydroxy, C1-C6Alkyl radical, C1-C6Haloalkyl, -OR5、(C1-C6Alkoxy group)r-O-C1-C6Haloalkyl, C1-6alkyl-NHR5、-N(R5)2、-NH(R5)-CO2H、C1-C6alkoxy-OCHO, -CO2R5、C1-C6alkyl-CO2R5、-CONHR5、-O-Y、-CONH(R5)-Y、CO-Y、C1-C14alkyl-Y, C1-C14alkoxy-Y and Y;
each Y is independently selected from C6-C10Aryl radical, C3-C12A cycloalkyl, a 5-to 10-membered heteroaryl, or a 3-to 12-membered heterocyclyl, wherein the heteroaryl or heterocyclyl includes 1-3 ring heteroatoms selected from O, N and S, wherein Y is optionally substituted with one to three substituents selected from: halogen, cyano, hydroxy, C1-C6Alkyl radical, C1-C6Alkoxy, -CO2H、-CO2C1-C4Alkyl and-O-C6-C10An aryl group;
m is 0, 1, 2 or 3;
q is 0, 1, 2 or 3 and if X is not-C (R)5)2-or 5 to 10 membered heteroaryl, then q is 1, 2 or 3; and
each of r and s is 0, 1, 2 or 3; and r + s is 1 to 6.
In some cases, the compound has the structure of one of the following formulas:
or a pharmaceutically acceptable salt thereof, wherein:
x is-C (R)5)2-、-N(R5)-、-NR5-SO2-or-O-;
each Q is independently N, CH or C, wherein indicates the point of attachment of the L group and only one Q is C; and
n is independently 1, 2, 3 or 4, and all remaining substituents are as described for one of formulas A, B or C.
In some embodiments, the compounds described herein may have one of the following formulas:
wherein R' is-H or C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, amino, CO2H、C1-C6Alkoxy radical, C1-C6Haloalkoxy, C1-C6Aminoalkoxy group, C1-C6Cyanoalkoxy group, C1-C6Hydroxyalkoxy and C2-C6An alkoxyalkoxy group.
In the embodiments of the formulae described herein, a side chain moiety L that is not shown attached to a particular position in the structure is understood to be attached at any open position.
In some embodiments, the compounds of formulae 7 to 13 do not comprise a side chain, i.e., L ═ H.
The side chain moiety L may have one of the following formulae:
-CH2CH2OCH2CH2OCH2CH2-T, wherein T is selected from the group consisting of: OH, OR5、NHR5、CO2R5、CO2NHR5、C1-6alkyl-CO2R5、C1-6alkyl-NHR5、C6-C10Aryl, 5-to 10-membered heteroaryl, C1-8Alkyl and C3-8A cycloalkyl group;
-C1-8alkoxy-C1-8alkoxy-U;
-C1-8alkoxy-C1-8alkoxy-C1-8An alkoxy group-U, which is a cyclic alkoxy group,
-C1-8alkoxy-aryl/heteroaryl-C1-8alkyl-C1-8Alkoxy radical
-C6-13alkyl-U;
-C1-8alkyl-C1-8An alkoxy group-U, which is a cyclic alkoxy group,
-C1-8alkyl-C1-8alkoxy-C1-8An alkoxy group-U, which is a cyclic alkoxy group,
-C1-8alkyl-C1-8alkoxy-C1-8alkoxy-C1-8An alkoxy group-U, which is a cyclic alkoxy group,
-C1-8alkyl-aryl/heteroaryl-C1-8alkoxy-C 1-8An alkoxy group-U, which is a cyclic alkoxy group,
-C1-8alkyl-aryl/heteroaryl-C1-8alkyl-C1-8alkoxy-C1-8An alkoxy group-U, which is a cyclic alkoxy group,
-C1-8alkyl-aryl/heteroaryl-C1-8alkyl-C1-8alkoxy-C1-8An alkyl group-U, wherein the alkyl group is,
-C1-8alkyl-aryl/heteroaryl-C1-8alkyl-C1-8An alkoxy group-U, which is a cyclic alkoxy group,
-C1-8alkyl-C1-8alkoxy-aryl/heteroaryl-C1-8An alkoxy-aryl group, which is a cyclic alkyl group,
-C1-8alkyl-C1-8alkoxy-aryl/heteroaryl-C1-8An alkyl group-U, wherein the alkyl group is,
-C1-8alkyl-C1-8alkoxy-aryl/heteroaryl-C1-8An alkyl-alkoxy-U, in which,
-C1-8alkyl- (optionally substituted) phenoxy;
-C1-8alkyl-C1-8Alkoxy- (optionally substituted) phenoxy;
-C1-8alkyl-N (R)5)-C(O)-C1-8alkyl-C1-8alkoxy-U;
-C1-8alkyl-O-C (O) N (R)5)-C1-8alkyl-U;
-C1-8alkyl-aryl/heteroaryl-C1-8alkoxy-U;
-aryl/heteroaryl-U;
-aryl/heteroaryl-C1-8alkoxy-U;
-aryl/heteroaryl-C1-8alkoxy-C1-8alkyl-U;
-aryl/heteroaryl-C1-8alkoxy-C1-8alkoxy-U;
aryl/heteroarylradical-C1-8alkyl-C1-8alkoxy-C1-8alkoxy-U; or
-aryl/heteroaryl-C1-8alkoxy-C1-8alkoxy-U;
u is selected from the group consisting of: H. c1-8Alkoxy radical, C6-C10Aryloxy group, OR5、NHR5、CO2R5、C(O)NHR5、C1-6alkyl-CO2R5、C1-6alkyl-NHR5、C6-C10Aryl radical, C6-C10aryl-C1-6Alkyl radical, C1-6alkyl-C6-C10Aryl, 3-to 12-membered heterocycloalkyl, 5-to 10-membered heteroaryl, C1-8Alkyl radical, C1-6Haloalkyl and C3-8Cycloalkyl, wherein the heterocycloalkyl and heteroaryl have 1 to 3 ring heteroatoms selected from N, O and S,
Wherein the alkyl, alkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, and cycloalkyl moieties may be optionally substituted with one to three (e.g., 1, 2, or 3) substituents selected from the group consisting of: hydroxy, C1-6Hydroxyalkyl radical, C1-6Alkoxy radical, C2-8Alkoxyalkyl group, C2-8Alkyl alkoxy radical, C1-8Alkoxycarbonyl group, C1-8Alkyl, arylalkoxycarbonyl, C2-6Alkenyl radical, C2-6Alkynyl, CO2H. Halogen, C1-6Haloalkyl, N3Cyano, N (R')2、SR'、-C(O)NHR'、OCOR'、OC(O)NHR'、N(CO)R'、N(CO)OR'、N(CO)COR'、SCOR'、S(O)2NR'2、S(O)2R ', wherein each R' is independently H, C1-6Alkyl radical, C1-6Haloalkyl, C1-6Alkoxy radical, C2-6Alkenyl radical, C2-6Alkynyl, C3-6Cycloalkyl, heterocyclyl, aryl, aryloxy, heteroaryl, heteroaryloxy, alkylaryl, arylalkyl, heteroarylalkyl, or alkylheteroaryl, and said heteroaryl having 1 to 3 ring heteroatoms selected from N, O and S.
Representative side chain moieties L may comprise side chains S1 to S57:
in some embodiments, Z is for any of formulas A, B or C, or formulas 1 through 131And R3Is F or Cl, and R1、R2、Z2And Z3Is H. In some aspects, Z1And R3One or both of which are F.
In some embodiments, for formula A, B or C, or any one of formulae 1 to 13, or a pharmaceutically acceptable salt thereof: r1is-H; r 2is-H; r3is-H, -F, -Cl, C1-4Alkyl or C1-4A haloalkyl group; z1is-H, -F or-Cl; z2is-H; z3is-H; each R8Independently is-H, halogen, hydroxy, C1-C4Alkyl radical, C1-C4Haloalkyl, C1-C4Hydroxyalkyl radical, C2-C4Alkoxyalkyl or-O (C)1-C4Alkyl groups); and R is9Independently is-H or C1-C4An alkyl group.
In some embodiments, the compounds described herein may have the formula:
wherein Het/Ar is C6-C10Aryl or 5 to 10 membered heteroaryl; l is C1-C14alkyl-NR5C(O)-C1-C14Alkyl-RzOr C1-C14alkyl-OC (O) NH-C1-C14alkyl-Rz(ii) a And R iszIs H, C1-C6Alkyl or C1-C6alkyl-CO2R5And said alkyl is optionally substituted with one or more (e.g. 1, 2 or 3) of: hydroxy, halogen, C1-C6Alkoxy radical, C1-C6Haloalkoxy, NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO-C1-C4Alkyl, -CO2H and-CO2C1-C4An alkyl group.
In some embodiments, Het/Ar is pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, furanyl, pyranyl, thienyl, benzimidazolyl, benzothienyl, benzofuranylbenzotriazolyl, benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, indolyl, isoindolyl, purinyl, phenyl, or naphthyl. In some embodiments, Het/Ar is pyridinyl or phenyl. In some embodiments, Het/Ar is pyridinyl.
In some embodiments, L is C1-C14alkyl-OC (O) NH-C1-C14alkyl-Rz. In some embodiments, L is C1-C6alkyl-OC (O) NH-C1-C6alkyl-Rz。
In some embodiments, the compounds described herein may have the formula:
wherein R' is-H or C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, amino, carboxy, C1-C6Alkoxy radical, C1-C6Haloalkoxy, C1-C6Aminoalkoxy group, C1-C6Cyanoalkoxy group, C1-C6Hydroxyalkoxy and C2-C6An alkoxyalkoxy group.
In some embodiments, the compounds described herein may have the formula:
wherein R' is-H or C1-C6Alkyl optionally substituted with one or more (e.g. 1, 2 or 3) substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, amino, carboxy, C1-C6Alkoxy radical, C1-C6Haloalkoxy, C1-C6Aminoalkoxy group, C1-C6Cyanoalkoxy group, C1-C6Hydroxyalkoxy and C2-C6An alkoxyalkoxy group.
It will be appreciated that the values of each variable are selected such that stable or chemically feasible compounds are formed.
The specific compounds contemplated include those in the table below. Compounds that exhibit a particular stereogenic center indicate at least relative stereoisomerism, and may refer to absolute stereoisomerism. Compounds having a chiral center that do not indicate a particular stereoisomer indicate a mixture of stereogenic centers at that chiral center.
The compound may be a compound listed in table a or a pharmaceutically acceptable salt thereof.
TABLE A
Application method
The compounds described herein, or pharmaceutically acceptable salts thereof, can be used to reduce viral titer in a biological sample (e.g., infected cell culture) or in a human (e.g., pulmonary viral titer in a patient).
As used herein, the terms "influenza virus-mediated symptom," "influenza infection," or "influenza" are used interchangeably to mean a disease caused by an influenza virus infection.
Influenza is an infectious disease caused by influenza virus that affects birds and mammals. Influenza viruses are RNA viruses of the orthomyxoviridae family, which includes five genera: influenza A virus, influenza B virus, influenza C virus, infectious salmon anemia virus and Togavirus. Influenza a virus genus has one species, influenza a virus, which can be subdivided into different serotypes based on antibody responses to these viruses: H1N1, H2N2, H3N2, H5N1, H7N7, H1N2, H9N2, H7N2, H7N3, H7N9, and H10N 7. Influenza B virus has one species, influenza B virus. Influenza B almost completely infects humans and is less prevalent than influenza a. Influenza C virus has one species, influenza C virus, which infects humans and pigs and can cause severe disease and local epidemics. However, influenza C viruses are less prevalent than other types and appear to often cause mild disease in children.
In some embodiments, the influenza or influenza virus is associated with an influenza a or B virus. In some embodiments, the influenza or influenza virus is associated with influenza a virus. In some particular embodiments, the influenza a virus is H1N1, H2N2, H3N2, H7N9, or H5N 1.
In humans, common symptoms of influenza are chills, fever, pharyngitis, muscle aches, severe headaches, coughing, weakness and general malaise. In more severe cases, influenza causes pneumonia, which can be fatal, especially for young children and the elderly. Although it is often confused with the cold, influenza is a much more serious disease and is caused by different virus types. Influenza can cause nausea and vomiting, particularly in children, but these symptoms are more characteristic of unrelated gastroenteritis, sometimes referred to as "gastric flu" or "24 hour flu".
Symptoms of influenza can appear quite suddenly one to two days after infection. Typically, the first symptom is chills or intolerance of cold, but fever is also common early in the infection, with body temperatures ranging from 38 ℃ to 39 ℃ (about 100 ° f to 103 ° f). Many people are so ill that they are bedridden for several days with general pain and discomfort that is severe in their backs and legs. Symptoms of influenza may include: general pain (especially in joints and throat), extreme cold and fever, weakness, headache, eye irritation tearing, eye redness, skin (especially facial) redness, mouth redness, throat redness and nasal redness, abdominal pain (children with influenza type B). Symptoms of influenza are not specific, but are superimposed on many pathogens ("influenza-like diseases"). Often, laboratory data is required to confirm the diagnosis.
The terms "disease," "disorder," and "symptom" are used interchangeably herein to refer to an influenza virus-mediated medical or pathological condition.
As used herein, the terms "individual" and "patient" are used interchangeably. The terms "subject" and "patient" refer to an animal (e.g., a bird such as a chicken, quail or turkey, or a mammal), specifically, "mammal" includes non-primates (e.g., cows, pigs, horses, sheep, rabbits, guinea pigs, rats, cats, dogs, and mice) and primates (e.g., monkeys, chimpanzees, and humans), and more specifically, humans. In some embodiments, the subject is a non-human animal, such as a farm animal (e.g., a horse, cow, pig, or sheep), or a pet (e.g., a dog, cat, guinea pig, or rabbit). In a preferred embodiment, the subject is a "human".
As used herein, the term "biological sample" includes, but is not limited to, cell cultures or extracts thereof; biopsy material obtained from a mammal or an extract thereof; blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.
As used herein, the term "inhibiting replication of influenza virus" encompasses both reducing the amount of viral replication (e.g., by at least 10%) and completely inhibiting viral replication (i.e., 100% reducing the amount of viral replication). In some embodiments, replication of the influenza virus is inhibited by at least 50%, at least 65%, at least 75%, at least 85%, at least 90%, or at least 95%.
Influenza virus replication can be measured by any suitable method known in the art. For example, the influenza virus titer of a biological sample (e.g., infected cell culture) or a human (e.g., a patient's lung virus titer) can be measured. More specifically, for cell-based assays, in each case where cells are cultured in vitro, virus is added to the culture in the presence or absence of the test agent, and the virus-dependent endpoints are assessed after a suitable length of time. For a typical assay, Madin-Darby canine kidney cells (MDCK) and standard tissue cultures appropriate for the influenza strain A/Puerto Rico/8/34 can be used. The first type of cell analysis that can be used depends on the death of the infected target cells, a process known as cytopathic effect (CPE), where viral infection causes depletion of the cellular source and eventual lysis of the cells. In the first type of cell assay, a small fraction of the cells in the wells of a microtiter plate are infected (typically 1/10-1/1000), the virus is subjected to several rounds of replication in 48-72 hours, and the amount of cell death is then measured using a reduction in cellular ATP content compared to uninfected controls. The second type of cellular analysis that can be used depends on the proliferation of virus-specific RNA molecules in the infected cells, where the RNA content is directly measured using the branched-chain DNA hybridization method (bDNA). In the second type of cell analysis, a lower number of cells are initially infected in the wells of a microtiter plate, the virus is replicated in the infected cells and spread to additional rounds of cells, which are then lysed and the viral RNA content is measured. This assay is usually stopped early after 18 to 36 hours, when all target cells are still viable. Viral RNA is quantified by hybridizing to specific oligonucleotide probes immobilized to the wells of the assay plate, followed by amplification of the signal by hybridization with additional probes linked to a reporter enzyme.
As used herein, "viral titer" or "titer" is a measure of viral concentration.Titer testing can employ serial dilutions to obtain approximate quantitative information from analytical procedures that themselves only evaluate as positive or negative. Titer corresponds to the highest dilution factor that still produced a positive reading; for example, positive reads in the first 8 serial two-fold dilutions converted to titers of 1: 256. A specific example is virus titer. To determine titre, several dilutions will be made, e.g. 10-1、10-2、10-3、……、10-8. The lowest concentration of virus that still infects cells is the virus titer.
As used herein, the term "treatment" refers to both therapeutic and prophylactic treatment. For example, therapeutic treatment includes reducing or ameliorating the progression, severity, and/or duration of an influenza virus-mediated condition, or ameliorating one or more symptoms (in particular, one or more discernible symptoms) of an influenza virus-mediated condition, by administering one or more therapies (e.g., one or more therapeutic agents, such as a compound or composition of the invention). In particular embodiments, the therapeutic treatment comprises ameliorating at least one measurable physical parameter of an influenza virus-mediated condition. In other embodiments, the therapeutic treatment comprises physically inhibiting the progression of the influenza virus-mediated condition, for example, by stabilizing a discernible symptom, physiologically, for example, by stabilizing a physical parameter, or both. In other embodiments, the therapeutic treatment comprises reducing or stabilizing an influenza virus-mediated infection. Antiviral drugs can be used in the community setting to treat people already with influenza, to reduce the severity of symptoms and to reduce their days of illness.
The term "chemotherapy" refers to the treatment of a disorder or disease with a drug, such as a small molecule drug (rather than a "vaccine").
As used herein, the terms "prevention", "prophylactic use" and "prophylactic treatment" refer to any medical or public health procedure aimed at preventing, rather than treating or curing, a disease. As used herein, the term "preventing" refers to reducing the risk of developing or manifesting an established condition, or reducing or inhibiting the recurrence of or the condition in an individual who is not ill but who has become or may be near to becoming a patient-carrying person. The term "chemoprevention" refers to the treatment of a disorder or disease with a drug, such as a small molecule drug (rather than a "vaccine").
As used herein, prophylactic use includes use in situations where outbreaks have been detected to prevent contagious infection or spread in locations where a large number of people in close contact with each other, such as hospital wards, day care centers, prisons, nursing centers, etc., are at high risk of serious influenza complications. It also includes use in people who need protection from influenza, but do not obtain protection after vaccination (e.g. due to a weaker immune system), or when the vaccine is not available for it or when it is not available due to side effects. It also encompasses use within two weeks after vaccination or during any period after vaccination but before the vaccine is effective. Prophylactic use may also include treating persons who are not suffering from influenza or are not considered to have a high risk of complications, to reduce the chance of contracting the influenza and transmitting it to persons at high risk of being in close contact therewith (e.g., healthcare workers, nursing center workers, etc.).
As used herein and in accordance with the american centers for Disease Control (US CDC), an influenza "outbreak" is defined as a sharp increase in the Acute Febrile Respiratory Illness (AFRI) occurring within a period of 48 to 72 hours in a population in close proximity to each other (e.g., in the same area of a assisted living facility, in the same household, etc.) relative to the normal prior art rate or when any individual in the population analyzed tests positive for influenza. One instance of influenza identified by any test method is considered an outbreak.
As used herein, "index case", "primary case" or "patient zero" is the first patient in the population sample of the epidemiological study. The indicated case was the first patient to indicate the presence of an outbreak. Earlier cases may be found and labeled as primary, secondary, tertiary, etc.
In some embodiments, the methods of the invention are prophylactic or preventative measures against patients (particularly humans) susceptible to complications caused by infection with influenza virus. The prophylactic methods described herein can be used in situations where it is confirmed that an indication of a case or outbreak is present to prevent the spread of infection in the rest of the community or population.
In some embodiments, the methods of the invention are applied as a prophylactic measure to members of a community or population, particularly humans, to prevent the spread of infection.
As used herein, "effective amount" refers to an amount sufficient to elicit a desired biological response. In the present invention, the desired biological response is to inhibit replication of influenza virus, to reduce the amount of influenza virus or to reduce or ameliorate the severity, duration, progression or onset of influenza virus infection, to prevent the acceleration of influenza virus infection, to prevent the recurrence, manifestation, onset or progression of symptoms associated with influenza virus infection, or to enhance or improve the prophylactic or therapeutic effect of another therapy used against influenza infection. The precise amount of the compound to be administered to an individual will depend on the mode of administration, the type and severity of the infection and the characteristics of the individual, such as general health, age, sex, weight and tolerance to drugs. One skilled in the art will be able to determine the appropriate dosage based on these and other factors. When co-administered with other antiviral agents, for example when co-administered with an anti-influenza drug, the effective amount of the second agent will depend on the type of drug used. A safe amount is an amount with minimal side effects, as can be readily determined by one skilled in the art. Suitable dosages for approved agents are known and can be adjusted by one of skill in the art depending on the condition of the individual, the type of condition being treated, and the amount of the compound described herein used. In the case where the amount is not explicitly labeled, a safe and effective amount should be assumed. For example, a compound described herein may be administered to a subject for therapeutic or prophylactic treatment at a dosage range of between about 0.01 and 100 mg/kg body weight/day.
As used herein, a "safe and effective amount" of a compound or composition described herein is an effective amount of the compound or composition that does not cause excessive or harmful side effects in a patient.
In general, the dosing regimen may be selected based on a variety of factors including: the condition being treated and the severity of the condition; the activity of the particular compound employed; the particular composition employed; the age, weight, general health, sex, and diet of the patient; the time of administration, route of administration, and rate of excretion of the particular compound employed; the renal and hepatic function of the individual; and the particular compound or salt employed, the duration of treatment; drugs used in combination or concomitantly with the particular compound employed; and similar factors well known in the medical arts. A safe and effective amount of a compound described herein needed for treating, preventing, inhibiting (in whole or in part), or arresting the progression of a disease can be readily determined and established by those skilled in the art.
The dosage of the compounds described herein may range between about 0.01 to about 100 mg/kg body weight/day, about 0.01 to about 50 mg/kg body weight/day, about 0.1 to about 50 mg/kg body weight/day, or about 1 to about 25 mg/kg body weight/day. It is understood that the total amount per day may be administered in a single dose, or may be administered in multiple doses, such as twice a day (e.g., every 12 hours), three times a day (e.g., every 8 hours), or four times a day (e.g., every 6 hours).
For therapeutic treatment, a compound described herein can be administered to a patient within, e.g., 48 hours (or within 40 hours, or less than 2 days, or less than 1.5 days or 24 hours) of the onset of symptoms (e.g., nasal congestion, sore throat, cough, pain, weakness, headache, and cold/night sweats). The therapeutic treatment may be for any suitable duration, e.g., for 5 days, 7 days, 10 days, 14 days, etc. For prophylactic treatment during a community outbreak, the compounds described herein can be administered to a patient, for example, within 2 days of the onset of symptoms in the indicated case, and can last for any suitable duration, e.g., for 7 days, 10 days, 14 days, 20 days, 28 days, 35 days, 42 days, etc.
Combination therapy
The compounds described herein, or pharmaceutically acceptable salts thereof, can be administered alone or in combination with additional suitable therapeutic agents, such as antiviral agents or vaccines. When combination therapy is employed, a safe and effective amount may be obtained using a first amount of a compound as disclosed herein, e.g., any of the compounds of formula A, B or C or any of formulae 1 to 13, or a pharmaceutically acceptable salt thereof, and a second amount of an additional suitable therapeutic agent (e.g., an antiviral agent or vaccine).
In some embodiments of the present disclosure, the compound described herein, or a pharmaceutically acceptable salt thereof, and the additional therapeutic agent are each administered in a safe and effective amount (i.e., each in an amount that is therapeutically effective if administered alone). In some embodiments, the compound and additional therapeutic agent are each administered in an amount that alone does not provide a therapeutic effect (lower than the therapeutic dose). In some embodiments, the compound may be administered in a safe and effective amount, while the additional therapeutic agent is administered at a lower than therapeutic dose. In some embodiments, the compound may be administered at a lower therapeutic dose while the additional therapeutic agent, e.g., a suitable cancer therapeutic agent, is administered in a safe and effective amount.
As used herein, the terms "combination therapy," "combination," and "co-administration" or "co-administration" are used interchangeably to refer to the use of more than one therapy (e.g., one or more prophylactic and/or therapeutic agents). The use of the terms does not limit the order of therapy (e.g., prophylactic and/or therapeutic) administered to an individual.
Co-administration may encompass administering the combined first and second amounts of the compound in a substantially simultaneous manner, such as in a single pharmaceutical composition, e.g., a capsule or tablet having a fixed ratio of the first and second amounts, or in a plurality of respective individual capsules or tablets. In addition, such co-administration may also encompass the use of each compound in a sequential manner, in either order.
In some embodiments, the present disclosure relates to a combination therapy for a method of inhibiting influenza virus replication or treating or preventing influenza virus infection in a patient using a compound or pharmaceutical composition of the invention in a biological sample or patient. Accordingly, the pharmaceutical compositions described herein also include those comprising an inhibitor of influenza virus replication as described herein in combination with an antiviral compound that exhibits anti-influenza virus activity.
Methods of use also include combinations of chemotherapy with a compound described herein or with a compound disclosed herein in combination with other antiviral agents and vaccination with an influenza vaccine.
Where co-administration involves administering a first amount of a compound as described herein and a second amount of an additional therapeutic agent separately, the compound and agent are administered in sufficient proximity to have the desired therapeutic effect. For example, the period of time between administrations that can produce a desired therapeutic effect can range from minutes to hours, and can be determined taking into account the characteristics of each compound (e.g., potency, solubility, bioavailability, plasma half-life, and kinetic profile). For example, a compound and a second therapeutic agent as described herein can be administered in any order within about 24 hours of each other, within about 16 hours of each other, within about 8 hours of each other, within about 4 hours of each other, within about 1 hour of each other, or within about 30 minutes of each other.
More specifically, a first therapy (e.g., a prophylactic or therapeutic agent, such as a compound of the invention) can be administered prior to (e.g., 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 before), concurrently with, or after (e.g., 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 after) administration of an additional therapeutic agent (e.g., an antiviral agent or an influenza vaccine) to a subject.
It is understood that a method of co-administering a first amount of a compound as described herein and a second amount of an additional therapeutic agent may produce an enhanced or synergistic therapeutic effect, where the combined effect is greater than the additive effect produced by the individual administration of the first amount of the compound as described herein and the second amount of the additional therapeutic agent.
As used herein, the term "synergistic" refers to a combination of a compound of the invention and another therapy (e.g., prophylactic or therapeutic agent) that is more effective than the additive effects of the therapies. The synergistic effect of a combination of therapies (e.g., a combination of prophylactic or therapeutic agents) may allow for the use of lower doses of one or more of the therapies and/or less frequent administration of the therapies to an individual. Being able to utilize lower doses of therapy (e.g., prophylactic or therapeutic agents) and/or less frequently administering the therapy can reduce the toxicity associated with administering the therapy to an individual without reducing the efficacy of the therapy in preventing, treating, or treating a condition. In addition, the synergistic effect may result in an improved efficacy of the agent in preventing, treating or treating the condition. Finally, the synergistic effect of a combination of therapies (e.g., a combination of prophylactic or therapeutic agents) can avoid or reduce adverse or undesirable side effects associated with the use of either therapy alone.
When combination therapy using the compounds of the present invention is combined with an influenza vaccine, the two therapeutic agents may be administered such that the period between administrations may be longer (e.g., days, weeks, or months).
The presence of a synergistic effect may be determined using suitable methods for assessing drug interactions. Suitable methods include, for example, the Sigmoid-Emax equation (Holford, N.H.G. and Scheiner, L.B., "clinical pharmacokinetics (Clin. Pharmacokinet) 6:429-453(1981)), the Loewe addition equation (Loewe, S and Muischnek, H.," Experimental pathology and pharmacology archives (Arch. Exp. Pathol. Pharmacol.) 114:313-326(1926)) and the efficacy equation (Chou, T.C. and Talalay, P., "Adv. enzyme Regulation progress (adv. enzyme Regul.)) 22:27-55 (1984)). The above mentioned programs can be applied to experimental data to generate corresponding charts that help assess the effect of drug combinations. The corresponding graphs associated with the above mentioned equations are the concentration-effect curve, the isobologram curve and the combined index curve, respectively.
Vaccine against influenza
The compounds described herein may be administered prophylactically in conjunction with an anti-influenza vaccine. These vaccines can be administered, for example, via subcutaneous or intranasal administration. Vaccination via subcutaneous injection typically induces IgG antibodies in serum with neutralizing activity and is extremely effective in preventing the progression of disease states to more severe disease states such as pneumonia and the like. IgA, however, is the major prophylactic component in the upper respiratory mucosa as the site of infection. IgA is also may be advantageous for administration of vaccines via the intranasal route, as it is not induced by subcutaneous administration.
Antiviral inhibitors
Various other compounds may be used in combination with the compounds described herein to treat or prevent influenza infection. Approved compounds include Neuraminidase (NA) inhibitors, ion channel (M2) inhibitors, polymerase (PB1) inhibitors, and other influenza antiviral agents.
There are three FDA-approved influenza antiviral drugs for use against influenza virus, comprising(zanamivir),(oseltamivir phosphate) and(peramivir)) earlier drugs, e.g.(amantadine) andrimantadine is approved for the treatment and prevention of influenza a.
Neuraminidase (NA) inhibitors are a class of drugs that block neuraminidase. It is commonly used as an antiviral drug because it blocks the function of viral neuraminidase of influenza virus by preventing propagation of influenza virus by budding from host cells. Representative neuraminidase inhibitors comprise oseltamivirZanamivirRanina mivirAnd peramivir
M2 inhibitors may also be used. The matrix-2 (M2) protein is a proton-selective ion channel protein and is entirely located in the viral envelope of influenza A virus.
The anti-influenza virus drug amantadine is a specific blocker of the M2H + channel. Amantadine (comprising amantadine and rimantadine) has been widely abandoned because of virus resistance, but combination therapy can mitigate the development of resistance to these agents, as viruses that become resistant to one active agent can still be treated by the other agents in the combination therapy.
Inhibitors of influenza RNA-dependent RNA polymerase (RdRp) include favipiravir (favipiravir) and the compounds described in PCT WO 2013/138236. Additional compounds disclosed in Muratore et al, Proc. Natl. Acad. Sci. USA (PNAS), 109(16), 6247-:
specific examples of compounds that may be co-administered with the compounds described herein include: neuraminidase inhibitors, e.g. oseltamivirAnd zanamivirBlockers of viral ion channels (M2 protein), e.g. amantadineAnd rimantadineAnd antiviral drugs described in WO 2003/015798, including T-705 in the development of Fushan Chemical of Japan (Toyama Chemical). (see also ruuta et al, Antiviral res.,82:95-102(2009)) in some embodiments, the compounds described herein may be co-administered with a traditional influenza vaccine.
The compounds described herein may be useful as inhibitors of influenza virus replication in biological samples or patients. These compounds may also be useful for reducing the amount of influenza virus (virus titer) in a biological sample or patient. It may also be suitable for the therapeutic and prophylactic treatment of infections caused by influenza viruses in biological samples or patients.
The present disclosure also provides methods of making the compounds described herein. In some embodiments, the methods involve preparing a compound represented by formulas 1-13, or a pharmaceutically acceptable salt thereof.
Preparation of Compounds disclosed herein
The synthesis of the compounds described herein may be simplified using one or more backbones. For example, the backbone of one of the following formulas may be reacted with a suitably functionalized pyrimidine ring to form a core structure comprising an azaindole attached to the pyrimidine ring.
The present disclosure also provides methods of making the compounds described herein. The compounds described herein, and the pharmaceutical salts and prodrugs thereof, all comprise a common core comprising an azaindole ring coupled to a pyrimidine ring. In some embodiments, these two ring systems with appropriate substitution patterns to provide the compounds described herein can be coupled using the chemistries described below.
In some embodiments, the method comprises the steps of: reacting a precursor A:
reaction with precursor B or B1:
to form a compound represented by formula I1 (an intermediate, or "I" of the compound of formula 1):
other salts besides potassium salts may be used and any of a variety of Leaving Groups (LG) may be used, including tosylate, 4-nitrobenzenesulfonate, bromobenzenesulfonate, mesylate, triflate, and the like.
Representative examples of precursors B and B1 include the following:
wherein Ts is tosylate, also known as "tosyl" or "tosylate".
The variables (Z) in these formulae1、Z2、Z3、R2、R3、R4X and L) are the same as the definitions provided in the section defining the compounds described herein, or may be protected forms of the functional groups defined by the variables or synthetic components of such groups, where the functional groups will be unstable under the reaction conditions described herein. Examples of protecting Groups are described in Greene, T.W., Wuts, P.G, "Protective Groups in Organic Synthesis", third edition, John Wiley&Sons, new york: 1999 (and other versions of the book), the entire contents of which are incorporated herein by reference.
Any suitable reaction conditions known in the art, for example in PCT WO 2005/095400 and PCT WO 2007/084557 for the coupling of dioxaborolanes with chloro-pyrimidines may be used beforeReaction between the entities (A) and (B or B1). For example, the reaction between precursors (a) and (B or B1) may be in the presence of Pd (PPh)3)4The process is carried out as follows. Certain exemplary conditions are described in the examples section of this disclosure.
Replacing a leaving group on a compound of formula I1 to obtain a compound of formula 1:
wherein R is1As described above.
In some embodiments of the synthesis of compounds of formula I1, the synthesis comprises contacting the precursor C1 or C2 with an appropriately substituted cyclohexylamine (i.e., N (R) 4) C (o) X-L substitution) to form a compound represented by structural formula I1, as shown in scheme B below:
procedure B
In some embodiments, the leaving group is a tosylate group, and the tosylate group is "demethylbenzenesulfonated" to yield an intermediate of formula (IA). Any suitable conditions known in the art for deprotecting the Ts group may be employed. Certain exemplary conditions are described in the examples. The demethylation benzenesulfonylation can yield a compound of formula (IA) wherein R1is-H. If necessary, R1The positions may be alkylated by any suitable method known in the art to form compounds of formula (IA) wherein R1Is C1-6An alkyl group.
Precursors (a), (B1), (C1), (C2) and appropriately substituted cyclohexylamine may be prepared by any suitable method known in the art. Specific exemplary synthetic methods are described in the following examples.
Similar chemistry can be used to prepare compounds of formulae 2 to 11. A list of suitable precursors for use in scheme B is provided below:
alternatively, we can use compounds of the structure of intermediate Int-11 to prepare compounds of the structure of intermediate Int-12
From Int-12, we can prepare compounds of formula 1 (typically involving the use of trialkylamines as proton sponges) by reaction with compounds of formula L-X-C (O) -Cl under appropriate amidation conditions.
From Int-12, we can prepare compounds of formulae 2 to 3 by reaction with appropriately substituted azacyclopentane-C (O) Cl or azacyclohexane-C (O) Cl compounds under appropriate amidation conditions.
From Int-12, we can prepare compounds of formulas 4 to 5 by reaction with appropriately substituted tetrahydroisoquinoline-c (o) Cl under appropriate amidation conditions.
From Int-12, we can prepare compounds of formulas 8 to 9 by reaction with appropriately substituted pyridine-C (O) Cl, pyrimidine-C (O) Cl, or pyrazine-C (O) Cl under appropriate amidation conditions.
From Int-12, we can prepare compounds of formulas 10 to 13 by reaction with an appropriately substituted azanaphthalene-C (O) Cl under appropriate amidation conditions.
Chiral separation
The compounds described herein may have asymmetric centers and exist as racemates, racemic mixtures, individual diastereomers or enantiomers, with all isomeric forms being embraced by the present invention. The compounds described herein having chiral centers may exist and be isolated in optically active and racemic forms. Some compounds may exhibit polymorphism. The present disclosure encompasses racemic, optically-active, polymorphic, or stereoisomeric forms, or mixtures thereof, of the compounds described herein having useful properties described herein. Optically active forms can be prepared, for example, by resolution of the racemic form by recrystallization techniques, by synthesis from optically active starting materials, by chiral synthesis, or by chromatographic separation using a chiral stationary phase, or by enzymatic resolution. We can purify the corresponding compound, followed by derivatizing the compound to form the compounds described herein or purifying the compound itself.
Optically active forms of the compounds can be prepared using any method known in the art, including, but not limited to, by resolution of the racemic form using recrystallization techniques, by synthesis from optically active starting materials, by chiral synthesis, or by chromatographic separation using a chiral stationary phase.
Examples of the method for obtaining an optically active substance include at least the following.
i)Physical separation of the crystals:techniques whereby the macroscopic crystallization of individual enantiomers is separated manually. This technique can be used if crystals of the respective enantiomers exist, i.e., the substance is an agglomerate and the crystals are visually distinct;
ii)and (3) simultaneous crystallization:the technique of crystallizing individual enantiomers separately from a solution of the racemate is only possible when the racemate is a solid agglomerate;
iii)enzymatic resolution:whereby racemates are partially or completely separated by virtue of the difference in the reaction rate of the enantiomers with the enzyme;
iv)enzymatic asymmetric synthesis:at least one step of the synthesis uses synthetic techniques of enzymatic reactions to obtain enantiomerically pure or enriched synthetic precursors of the desired enantiomer;
v)chemical asymmetric synthesis:synthetic techniques whereby the desired enantiomer is synthesized from an achiral precursor under conditions that produce asymmetry (i.e., chirality) in the product, can be achieved using chiral catalysts or chiral auxiliary agents;
vi)Diastereomer separation:whereby the racemic compound and the individual pairsTechniques for the reaction of enantiomerically pure reagents (chiral auxiliary) for the conversion of enantiomers to diastereomers. The resulting diastereoisomers are then separated by chromatography or crystallization by virtue of their now more pronounced structural differences and subsequent removal of the chiral auxiliary to give the desired enantiomer;
vii)first and second stages of asymmetric transformation:techniques in which diastereoisomeric equilibrium from a racemate is preferentially resolved to give diastereoisomers from a desired enantiomer, or preferential crystallization of diastereoisomers from a desired enantiomer interferes with equilibrium so that ultimately substantially all of the material is converted to a crystalline diastereoisomer from a desired enantiomer. Followed by liberation of the desired enantiomer from the diastereomer;
viii)and (3) kinetic resolution:this technique refers to the partial or complete resolution of racemates (or further resolution of partially resolved compounds) achieved by virtue of the unequal reaction rates of the enantiomers with chiral non-racemic reagents or catalysts under kinetic conditions;
ix)enantiospecific synthesis from non-racemic precursors:synthetic techniques whereby the desired enantiomer is obtained from a non-chiral starting material and wherein the stereochemical integrity is not or only minimally impaired during the synthesis;
x)Chiral liquid chromatography:techniques whereby the enantiomers of a racemate are separated by virtue of their different interactions with a stationary phase, including (but not limited to) via chiral HPLC, in a liquid mobile phase. The stationary phase may be made of a chiral substance or the mobile phase may contain another chiral substance to cause different interactions;
xi)chiral gas chromatography:techniques whereby the racemate is volatilized and the enantiomers are separated by virtue of their different interactions in the gas mobile phase with a column containing a fixed non-racemic chiral adsorbent phase;
xii)extraction with a chiral solvent:whereby the enantiomers are separated by preferential dissolution of one enantiomer in a particular chiral solvent;
xiii)transport across the chiral membrane:thereby contacting the racemate with the film barrier. The barrier typically separates two miscible fluids, one containing the racemate, and the driving force (e.g., concentration or pressure differential) causes preferential transport across the membrane barrier. The separation is carried out by the non-racemic chiral nature of the membrane, which allows only one enantiomer of the racemate to pass through.
In one embodiment, chiral chromatography, including but not limited to simulated moving bed chromatography, is used. A variety of chiral stationary phases are commercially available.
Definitions and general terms
For the purposes of this disclosure, chemical Elements are identified according to the Periodic Table of the Elements (CAS version, Handbook of Chemistry and Physics), 75 th edition. In addition, general principles of Organic Chemistry are described in "Organic Chemistry" (in Thomas Sorrell, "University Science Books, Sausaltito"), 1999 and "March's Advanced Organic Chemistry" (in March's Advanced Organic Chemistry), 5 th edition, editor: smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are incorporated herein by reference.
As described herein, the compounds described herein can be optionally substituted with one or more substituents as generally shown below or exemplified by the particular classes, subclasses, and species described herein. It will be appreciated that the phrase "optionally substituted" may be used interchangeably with the phrase "substituted or unsubstituted. In general, the term "substituted," whether or not preceded by the term "optionally," means that one or more hydrogen groups in a given structure are replaced with a group of a particular substituent. Unless otherwise specified, an optionally substituted group may have a substituent at each substitutable position of the group. When more than one position in a given structure may be substituted with more than one substituent selected from a particular group, the substituents may be the same or different at each position. When the term "optionally substituted" precedes a list, the term refers to all subsequent substitutable groups in the list. If the substituents or structures are not present Identified or defined as "optionally substituted," then the substituent or structure is unsubstituted. For example, if X is optionally substituted C1-C3Alkyl or phenyl; then X may be optionally substituted C1-C3Alkyl or optionally substituted phenyl. Likewise, if the term "optionally substituted" follows a list, the term also refers to all substitutable groups in the previous list, unless otherwise indicated. For example: if X is C1-C3Alkyl or phenyl, wherein X is optionally and independently via JXSubstituted, then C1-C3Alkyl and phenyl groups may optionally be via JXAnd (4) substitution. As will be apparent to those skilled in the art, e.g., H, halogen, NO2、CN、NH2OH or OCF3Is an unsubstituted group.
As used herein, the phrase "at most" refers to zero or any integer equal to or less than the number following the phrase. For example, "at most 3" means any of 0, 1, 2, and 3. As described herein, the specified range of numbers of atoms includes any integer therein. For example, a group having 1-4 atoms can have 1, 2, 3, or 4 atoms.
As used herein, the phrase "substituted with one or more instances" indicates that the moiety referred to bears at least one substituent as defined herein, as permitted by the valence of the substituted moiety. For example, a 6-membered aryl ring (e.g., a phenyl ring) substituted with one or more instances of J may be substituted with any of 1, 2, 3, 4, 5, and 6J.
The choice of substituents and combinations of substituents contemplated herein are those that form stable or chemically feasible compounds. As used herein, the term "stable" means that the compound does not substantially change when subjected to conditions that allow its production, detection, and rather its recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, a stabilizing compound or chemically feasible compound is a compound that is not substantially altered when held at a temperature of 40 ℃ or less for at least one week in the absence of moisture or other chemical reaction conditions. Only those choices and combinations of substituents that result in stable structures are contemplated. Such selections and combinations will be apparent to those skilled in the art and may be determined without undue experimentation.
As used herein, the term "aliphatic group" or "aliphatic radical" means a straight (i.e., non-branched) or branched hydrocarbon chain that is fully saturated or contains one or more units that are unsaturated but not aromatic. Unless otherwise specified, aliphatic groups contain 1 to 20 aliphatic carbon atoms. In some embodiments, the aliphatic group contains 1 to 10 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1 to 8 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1 to 6 aliphatic carbon atoms, and in other embodiments, the aliphatic group contains 1 to 4 aliphatic carbon atoms. The aliphatic group may be a straight or branched, substituted or unsubstituted alkyl, alkenyl or alkynyl group. Specific examples include, but are not limited to, methyl, ethyl, isopropyl, n-propyl, sec-butyl, vinyl, n-butenyl, ethynyl, and tert-butyl, and acetylene.
As used herein, the term "alkyl" means a saturated straight or branched chain hydrocarbon. As used herein, the term "alkenyl" means a straight or branched chain hydrocarbon comprising one or more double bonds. As used herein, the term "alkynyl" means a straight or branched chain hydrocarbon including one or more triple bonds. As used herein, each of "alkyl", "alkenyl", or "alkynyl" may be optionally substituted as set forth below. In some embodiments, "alkyl" is C1-C14Alkyl radical, C1-C6Alkyl or C1-C4An alkyl group. In some embodiments, "alkenyl" is C2-C6Alkenyl or C2-C4An alkenyl group. In some embodiments, "alkynyl" is C2-C6Alkynyl or C2-C4Alkynyl.
The term "cycloaliphatic" (or "cycloalkyl", "carbocycle" or "carbocyclyl" or "carbocyclic") refers to a non-aromatic carbon-only ring system which may be saturated or contain one or more units of unsaturation, having from 3 to 14 ringsA carbon atom. In some embodiments, the number of carbon atoms is 3 to 12 (i.e., C)3-C12Cycloalkyl groups). In other embodiments, the number of carbon atoms is from 4 to 7. In other embodiments, the number of carbon atoms is 5 or 6. The term encompasses monocyclic, bicyclic or polycyclic fused, spiro or bridged carbocyclic ring systems. The term also encompasses polycyclic ring systems wherein a carbocyclic ring may be "fused" to one or more non-aromatic carbocyclic or heterocyclic rings or one or more aromatic rings or combinations thereof, wherein a group or point of attachment is on the carbocyclic ring. A "fused" bicyclic ring system includes two rings that share two adjacent ring atoms. The bridged bicyclic group includes two rings that share three or four adjacent ring atoms. Spiro bicyclic systems share a common ring atom. Examples of cycloaliphatic radicals include, but are not limited to, cycloalkyl and cycloalkenyl. Specific examples include, but are not limited to, cyclohexyl, cyclopropenyl, cyclobutyl, and cyclopropyl.
As used herein, the term "heterocycle" (or "heterocyclyl" or "heterocyclic" or "non-aromatic heterocycle") refers to a non-aromatic ring system that may be saturated or contain one or more units of unsaturation, having three to fourteen ring atoms, wherein one or more ring carbons are replaced with a heteroatom such as N, S or O. In some embodiments, the ring system may comprise 3 to 12 ring atoms (i.e., a 3-to 12-membered heterocyclyl). In some embodiments, the non-aromatic heterocycle includes up to three heteroatoms selected from N, S and O within the ring. In other embodiments, the non-aromatic heterocyclic ring includes up to two heteroatoms selected from N, S and O within the ring system. In other embodiments, the non-aromatic heterocyclic ring includes up to two heteroatoms selected from N and O within the ring system. The term encompasses monocyclic, bicyclic or polycyclic fused, spiro or bridged heterocyclic systems. The term also encompasses polycyclic ring systems wherein a heterocyclic ring may be "fused" to one or more non-aromatic carbocyclic or heterocyclic rings or one or more aromatic rings or combinations thereof, wherein the group or point of attachment is on the heterocyclic ring. Examples of heterocycles include (but are not limited to): piperidinyl, piperazinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, azepanyl, diazepanyl, triazacyclocycloheptyl, azocyclooctyl, diazacyclooctyl, triazacyclooctyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, oxazepinyl, oxazepanyl, thiazepanyl, thiazepinyl, benzimidazolonyl, tetrahydrofuranyl, tetrahydrothienyl, N-morpholinyl (including, for example, 3-N-morpholinyl, 4-N-morpholinyl, 2-N-thiomorpholinyl, 3-N-thiomorpholinyl, 4-N-thiomorpholinyl), 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 1-tetrahydropyriperazinyl, 2-tetrahydropyriperazinyl, diazepanyl, thiazepanyl, benzimidazolonyl, tetrahydrofuranyl, tetrahydrothienyl, N-morpholinyl (, 3-tetrahydropyrizinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, 5-pyrazolinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 2-thiazolinyl, 3-thiazolinyl, 4-thiazolinyl, 1-imidazopinyl, 2-imidazopinyl, 4-imidazopinyl, 5-imidazopinyl, indolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, benzodithiol-heterocyclopentyl, benzodithietanyl, 3- (1-alkyl) -benzimidazol-2-onyl and 1, 3-dihydro-imidazol-2-onyl.
As used herein, the term "aryl" refers to a monocyclic aromatic group, such as phenyl. Unless otherwise specified, an aryl group can have 6 to 14 carbon atoms in the ring, such as 6 to 10 carbon atoms in the ring (i.e., C)6-C10Aryl). Unless otherwise indicated, aryl groups may be unsubstituted or substituted with one or more, and especially one to four groups independently selected from, for example: halo, alkyl, alkenyl, OCF3、NO2CN, NC, OH, alkoxy, amino, CO2H、CO2Alkyl, aryl and heteroaryl. The aryl group can be independent (e.g., phenyl) or fused with another aryl group (e.g., naphthyl, anthracenyl), cycloalkyl (e.g., tetrahydronaphthyl), heterocycloalkyl, and/or heteroaryl. Exemplary aryl groups include, but are not limited to, phenyl, chlorophenyl, methylphenyl, methoxyphenyl, trifluoromethylphenyl, nitrophenyl, 2, 4-methoxychlorophenyl, 1-naphthyl, 2-naphthyl, 1-anthryl, 2-anthryl, indanyl, phthalimidyl, naphthalimide, phenanthridinyl, or tetrahydronaphthyl, and the like.
The terms "heteroaryl", "heteroaromatic", "heteroaryl ring", "heteroaryl group", "aromatic heterocycle" or "heteroaromatic group", used alone or as part of a larger moiety in "heteroaralkyl" or "heteroarylalkoxy", refer to heteroaromatic ring groups having five to fourteen members, including monocyclic heteroaromatic rings and polycyclic aromatic rings, wherein a monocyclic aromatic ring is fused to one or more other aromatic rings. Heteroaryl groups have one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur in the aromatic ring. As used herein, a heteroaryl ring can have 5 to 14 ring atoms, such as 5 to 10 ring atoms (i.e., a 5-to 10-membered heteroaryl). The term "heteroaryl" as used herein also includes within its scope groups in which an aromatic ring is "fused" to one or more non-aromatic rings (carbocyclic or heterocyclic), wherein the group or point of attachment is on the aromatic ring. As used herein, for example, a bicyclic 6.5 heteroaromatic ring is a six membered heteroaromatic ring fused to a second five membered ring, wherein the group or point of attachment is on the six membered ring. Examples of heteroaryl groups include pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, or thiadiazolyl, including, for example, 2-furyl, 3-furyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxadiazolyl, 5-oxadiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-pyrazolyl, 4-pyrazolyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 3-pyridazinyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-triazolyl, 5-triazolyl, tetrazolyl, 2-thienyl, 3-thienyl, carbazolyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl, benzotriazolyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, isoquinolyl, indolyl, isoindolyl, acridinyl, benzisoxazolyl, isothiazolyl, 1,2, 3-oxadiazolyl, 1,2, 5-oxadiazolyl, 1,2, 4-oxadiazolyl, 1,2, 3-triazolyl, 1,2, 3-thiadiazolyl, 1,3, 4-thiadiazolyl, 1,2, 5-thiadiazolyl, Purinyl, pyrazinyl, 1,3, 5-triazinyl, quinolinyl (e.g., 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl, 3-isoquinolinyl, or 4-isoquinolinyl).
As used herein, "ring," "cyclic," "cyclyl," or "cyclic moiety" includes mono-, di-, and tricyclic ring systems, bridged bicyclic ring systems, and spiro ring systems, including cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups, or combinations thereof, each of which has been previously defined.
As used herein, "bridged bicyclic ring system" refers to a bicyclic heterocyclic aliphatic ring system or bicyclic aliphatic ring system in which the rings are bridged. Examples of bridged bicyclic ring systems include, but are not limited to, adamantyl, norbornyl, bicyclo [3.2.1] octyl, bicyclo [2.2.2] octyl, bicyclo [3.3.1] nonyl, bicyclo [3.2.3] nonyl, 2-oxa-bicyclo [2.2.2] octyl, 1-aza-bicyclo [2.2.2] octyl, 3-aza-bicyclo [3.2.1] octyl and 2, 6-dioxa-tricyclo [3.3.1.03,7] nonyl. The bridged bicyclic ring system may be optionally substituted.
As used herein, the term "spiro" refers to a ring system having one atom (typically the fourth carbon) as the only common atom between two rings.
The term "ring atom" is an atom such as C, N, O or S, which is located in the ring of a cycloalkyl, heterocycloalkyl, aryl or heteroaryl group.
A "substitutable ring atom" is a ring carbon or nitrogen atom that may be bonded to a hydrogen atom but is bonded to a moiety other than a hydrogen atom. Thus, the term "substitutable ring atom" does not include a ring nitrogen or carbon atom that is common when two rings are fused. In addition, a "substitutable ring atom" does not contain a ring carbon or nitrogen atom when the structure depicts that it is already attached to a moiety other than hydrogen.
Unless stated otherwise herein, the term "heteroatom" means one or more of oxygen, sulfur, nitrogen, phosphorus, or a combination thereof.
As used herein, the term "aralkyl" refers to an aryl ring attached to an alkyl chain, and "aralkoxy" refers to an aryl ring attached to an alkoxy chain. Optionally substituted aralkyl groups may be substituted on the alkyl and aryl moieties. As used herein, unless otherwise specified, an optionally substituted aralkyl is optionally substituted on the aryl moiety.
In some implementationsIn the examples, the aliphatic or heteroaliphatic or non-aromatic heterocyclic ring may contain one or more substituents. Suitable substituents on the saturated carbon of the aliphatic or heteroaliphatic group or of the non-aromatic heterocycle are selected from those listed above, for example in the definition of J. Other suitable substituents include those listed as unsaturated carbons suitable for carbocyclic aryl or heteroaryl, and additionally include the following: two (R) ═ O, ═ S, ═ NNHR, ═ NN (R)2、＝NNHC(O)R*、＝NNHCO2(alkyl) ═ NNHSO2(alkyl) or ═ NR, where each R is independently selected from hydrogen or optionally substituted C1-6Aliphatic. Optional substituents on the aliphatic radical of R are selected from NH2、NH(C1-4Alkyl group), N (C)1-4Alkyl radical)2Halogen, C1-4Alkyl, OH, O (C) 1-4Alkyl), NO2、CN、CO2H、CO2(C1-4Alkyl), O (halo C)1-4Alkyl) or halo (C)1-4Alkyl) in which R is C1-4Each of the alkyl groups is unsubstituted.
In some embodiments, the optional substituents on the nitrogen of the non-aromatic heterocycle include those used above, for example, in the definition of J. Other suitable substituents include-R+、-N(R+)2、-C(O)R+、-CO2R+、-C(O)C(O)R+、-C(O)CH2C(O)R+、-SO2R+、-SO2N(R+)2、-C(＝S)N(R+)2、-C(＝NH)-N(R+)2or-NR + SO2R+(ii) a Wherein R is+Is hydrogen, optionally substituted C1-6Alkyl, optionally substituted phenyl, optionally substituted-O (Ph), optionally substituted-CH2(Ph), optionally substituted- (CH)2)1-2(Ph); optionally substituted-CH ═ CH (ph); or unsubstituted 5-to 6-membered heteroaryl or heterocycle having one to four ring heteroatoms independently selected from oxygen, nitrogen and sulfur, or two independently occurring R on the same or different substituents+And each R+The atoms to which the groups are bonded together form a 5-to 8-membered heterocyclic, aryl groupOr a heteroaryl ring or a 3-to 8-membered cycloalkyl ring, wherein the heteroaryl or heterocyclyl ring has 1 to 3 ring heteroatoms independently selected from nitrogen, oxygen, and sulfur. R+Is selected from NH or an optional substituent on the phenyl ring2、NH(C1-4Alkyl group), N (C)1-4Alkyl radical)2Halogen, C1-4Alkyl, OH, O (C)1-4Alkyl), NO2、CN、CO2H、CO2(C1-4Alkyl), O (halo C)1-4Alkyl) or halo (C) 1-4Alkyl) in which R is+C of (2)1-4Each of the alkyl groups is unsubstituted.
In some embodiments, an aryl or heteroaryl group can include one or more (e.g., 1,2, or 3) substituents. Suitable substituents include: halogen; -R °; -OR °; -SR °; 1, 2-methylenedioxy; 1, 2-ethylenedioxy; phenyl (Ph) optionally substituted with R °; -o (ph) optionally substituted with R °; optionally R DEG substituted- (CH)2)1-2 (Ph); -CH ═ CH (ph) optionally substituted with R °; -NO2；-CN；-N(R°)2；-NR°C(O)R°；-NR°C(S)R°)2；-NR°C(O)N(R°)2；-NR°C(S)N(R°)2；-NR°CO2R°；-NR°NR°C(O)R°；-NR°NR°C(O)N(R°)2；-NR°NR°CO2R°；-C(O)C(O)R°；-C(O)CH2C(O)R°；-CO2R°；-C(O)R°；-C(S)R°；-C(O)N(R°)2；-C(S)N(R°)2；-OC(O)N(R°)2；-OC(O)R°；-C(O)N(OR°)R°；-C(NOR°)R°；-S(O)2R°；-S(O)3R°；-SO2N(R°)2；-S(O)R°；-NR°SO2N(R°)2；-NR°SO2R°；-N(OR°)R°；-C(＝NH)-N(R°)2(ii) a Or- (CH)2)0-2NHC (O) R °; wherein each independently occurring R DEG is selected from hydrogen, optionally substituted C1-6Alkyl, unsubstituted 5-to 6-membered heteroaryl or heterocycle, phenyl, -O (Ph) or-CH2(Ph), or two independently occurring R.cndot on the same or different substituents, together with the atom to which each R.cndot.group is bonded, form a 5-to 8-membered heterocyclyl, aryl or heteroaryl ring or a 3-to 8-membered cycloalkyl ringWherein said heteroaryl or heterocyclyl ring has 1 to 3 ring heteroatoms independently selected from nitrogen, oxygen, and sulfur. The optional substituents on the aliphatic radical of R DEG are selected from NH2、NH(C1-4Alkyl group), N (C)1-4Alkyl radical)2Halogen, C1-4Alkyl, OH, O (C)1-4Alkyl), NO2、CN、CO2H、CO2(C1-4Alkyl), O (halo C)1-4Alkyl) or halo (C)1-4Alkyl), CHO, N (CO) (C) 1-4Alkyl group, C (O) N (C)1-4Alkyl) in which R.degree.C1-4Each of the alkyl groups is unsubstituted.
Non-aromatic nitrogen-containing heterocycles that are substituted on the ring nitrogen and attached to the rest of the molecule at a ring carbon atom are referred to as N-substituted. For example, an N-alkylpiperidinyl group is attached to the remainder of the molecule at two, three, or four positions of the piperidinyl ring and is substituted with an alkyl group at the ring nitrogen. Non-aromatic nitrogen-containing heterocycles such as pyrazinyl, substituted on the ring nitrogen and attached to the rest of the molecule at the second ring nitrogen atom, are referred to as N' substituted-N-heterocycles. For example, the N' -acyl N-pyrazinyl group is attached to the rest of the molecule at one ring nitrogen atom and is substituted with an acyl group at a second ring nitrogen atom.
As detailed above, in some embodiments, two independently occurring R ° (or R °)+Or any other variable similarly defined herein) may be taken together with the atom to which each variable is bound to form a 5-to 8-membered heterocyclyl, aryl or heteroaryl ring, or a 3-to 8-membered cycloalkyl ring. Two independently occurring R ° (or R)+Or any other variable similarly defined herein) along with the atoms to which each variable is bound include (but are not limited to) the following: a) two independently occurring R ° (or R) +Or any other variable similarly defined herein) is bound to the same atom and forms a ring with that atom, e.g., N (R °)2Wherein two occurrences of R ° together with the nitrogen atom form piperidin-1-yl, piperazin-1-yl, or morpholin-4-yl; and b) two independently occurring R ° (or R °)+Or any other variable similarly defined herein) are bound to different atoms and to both of those atomsTogether forming a ring, e.g. wherein the phenyl group is substituted by two occurrences of ORThese two occurrences of R ° together with the oxygen atom to which they are bound form a fused 6-membered oxygen containing ring:
it is understood that various other rings can occur at two independently occurring R ° (or R [ - ])+Or any other variable similarly defined herein) are formed together with the atom to which each variable is bound, and the examples detailed above are not intended to be limiting.
In some embodiments, the alkyl chain may optionally be interrupted with another atom or group. This means that the methylene unit of the alkyl chain is optionally replaced by said another atom or group. Examples of such atoms or groups include, but are not limited to, those listed in the definitions of the variables after each formula. For example, the methylene group in the side chain may be replaced by: -NR 4-、-O-、-S-、-CO2-、-OC(O)-、-C(O)CO-、-C(O)-、-C(O)NR4-、-C(＝N-CN)-、-NR4CO-、-NR4C(O)O-、-SO2NR4-、-NR4SO2-、-NR4C(O)NR4-、-OC(O)NR4-、-NR4SO2NR4-, -SO-or-SO2-, wherein R4As defined above.
As used herein, "amino" refers to-NRXRYWherein R isXAnd RYIs independently each-H, C1-C6Alkyl radical, C3-7Cycloalkyl radical, C6-10Aryl, 5-to 6-membered heteroaryl, or 4-to 7-membered heterocycle, each independently defined herein and optionally substituted. Suitable substituents for these groups include halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C6Alkyl), -N (C)1-C6Alkyl radical)2、C1-C6Alkyl radical、-O(C1-C6Alkyl), -C (O) OH, -C (O) O (C)1-C6Alkyl), -OC (O) (C)1-C6Alkyl), -NHC (O) (C)1-C6Alkyl), -NHC (O) O (C)1-C6Alkyl), -C (O) NH (C)1-C6Alkyl) and-C (O) N (C)1-C6Alkyl radical)2Wherein each of said alkyl groups is optionally and independently substituted with one or more substituents selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl) and-N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO (C)1-C4Alkyl), -CO2H、-CO2(C1-C4Alkyl) and C1-C4An alkoxy group. In some embodiments, RXAnd RYEach of which is independently-H, optionally substituted C1-6Alkyl or optionally substituted C3-8A cycloalkyl group. In some embodiments, RXAnd RYEach of which is independently-H or optionally substituted C1-6An alkyl group. In some embodiments, RXAnd RYEach of which is independently-H or C optionally substituted with one or more substituents selected from the group consisting of 1-6Alkyl groups: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO (C)1-C4Alkyl), -CO2H、-CO2(C1-C4Alkyl) and C1-C4An alkoxy group. Examples of amino groups include-NH2Alkylamino, dialkylamino or arylamino.
As used herein, "amido" when used terminally encompasses N (R)XRY) -C (O) -or RYC(O)-N(RX) -and when used internally encompasses-C (O) -N (R)X) -or-N (R)x) -C (O) -, wherein RXAnd RYAs defined above. Examples of amide groups include alkylamidesA group (e.g., alkylcarbonylamino, or alkylaminocarbonyl), (heterocycloaliphatic) amido, (heteroaralkyl) amido, (heteroaryl) amido, (heterocycloalkyl) alkylamido, arylamido, aralkylamido, (cycloalkyl) alkylamido, or cycloalkylamido. In some embodiments, the amide group is-NHC (O) (C)1-C6Alkyl), -N (C)1-C6Alkyl radical C (O) (C)1-C6Alkyl), -C (O) NH2、-C(O)NH(C1-C6Alkyl) or-C (O) NH (C)1-C6Alkyl radical)2Wherein each of said alkyl groups is optionally and independently substituted with one or more substituents selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C6Alkyl), -N (C)1-C6Alkyl radical)2、-OCO(C1-C4Alkyl), -CO (C)1-C4Alkyl), -CO 2H、-CO2(C1-C4Alkyl) and C1-C4An alkoxy group. In some embodiments, the amide group is-NHC (O) (C)1-C6Alkyl), -N (C)1-C6Alkyl radical C (O) (C)1-C6Alkyl), -C (O) NH2、-C(O)NH(C1-C6Alkyl) or-C (O) NH (C)1-C6Alkyl radical)2Wherein each of said alkyl groups is optionally and independently substituted with one or more substituents selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C6Alkyl), -N (C)1-C6Alkyl radical)2、-OCO(C1-C4Alkyl), -CO (C)1-C4Alkyl), -CO2H、-CO2(C1-C4Alkyl) and C1-C4An alkoxy group.
As used herein, "acyl" refers to formyl or RX-C (O) - (e.g. -alkyl-C (O) -, also known as "alkylcarbonyl"), wherein R has been previously definedXAnd "alkyl". Acetyl and pivaloyl are examples of acyl groups.
As used herein, "carboxy" when used as a terminal group refers to-COOH, -COORX、-OC(O)H、-OC(O)RX(ii) a Or when used as an internal group means-OC (O) -or-C (O) O-, wherein RXAs defined above.
As used herein, "alkoxycarbonyl," encompassed by the term carboxy, used alone or in combination with another group, refers to a group such as (alkyl-O) -c (O) -.
As used herein, "carbonyl" refers to-C (O) -.
As used herein, "oxo" refers to ═ O.
As used herein, the term "alkoxy" or "alkylthio" as used herein refers to an alkyl group as previously defined attached to the molecule via an oxygen ("alkoxy", e.g., -O-alkyl) or sulfur ("alkylthio", e.g., -S-alkyl) atom.
As used herein, the terms "halo", and "halo" mean F, Cl, Br, or I.
As used herein, the term "cyano" or "nitrile" refers to — CN.
The terms "alkoxyalkyl", "alkoxyalkenyl", "alkoxyaliphatic" and "alkoxyalkoxy" mean an alkyl, alkenyl, aliphatic or alkoxy group that may be optionally substituted with one or more alkoxy groups.
The terms "haloalkyl", "haloalkenyl", "haloaliphatic" and "haloalkoxy" mean alkyl, alkenyl, aliphatic or alkoxy groups which may optionally be substituted with one or more halogen atoms. The term includes perfluoroalkyl groups, such as-CF3and-CF2CF3。
The terms "cyanoalkyl", "cyanoalkenyl", "cyanoaliphatic" and "cyanoalkoxy" mean an alkyl, alkenyl, aliphatic or alkoxy group which may be optionally substituted with one or more cyano groups. In some embodiments, cyanoalkyl is (NC) -alkyl-.
The terms "aminoalkyl", "aminoalkenyl" and "aminoalkoxy" mean an alkyl, alkenyl or alkoxy group optionally substituted with one or more amino groups, wherein amino is as aboveAs defined. In some embodiments, C 1-C6Alkyl via one or more-NH2And (4) substituting the group. In some embodiments, aminoalkyl refers to the structure (R)XRY) N-alkyl-, wherein RXAnd RYEach is independently as defined above. In some particular embodiments, aminoalkyl is substituted with one or more-NH2Radical substituted C1-C6An alkyl group. In some particular embodiments, aminoalkenyl is substituted with one or more-NH2Radical substituted C1-C6An alkenyl group. In some embodiments, the aminoalkoxy group is — O (C)1-C6Alkyl) in which the alkyl group is substituted with one or more-NH groups2And (4) substituting the group.
The terms "hydroxyalkyl" and "hydroxyalkoxy" mean an alkyl or alkoxy group that may be optionally substituted with one or more-OH groups.
The terms "alkoxyalkyl" and "alkoxyalkoxy" mean an alkyl or alkoxy group that may be optionally substituted with one or more alkoxy groups. For example, "alkoxyalkyl" refers to an alkyl group, such as (alkyl-O) -alkyl-, wherein alkyl is as defined above.
The term "carboxyalkyl" means an alkyl group substituted with one or more carboxy groups, wherein alkyl and carboxy are as defined above.
In some embodiments, each of the amino groups mentioned in the description for a variable of formula (e.g., formula A, B or C or formulae 1 to 13) disclosed herein is independently-NH 2、-NH(C1-C6Alkyl), -NH (C)3-C6Cycloalkyl), -N (C)1-C6Alkyl radical)2or-N (C)1-C6Alkyl) (C3-C6Cycloalkyl, wherein the alkyl and cycloalkyl are each optionally and independently substituted with one or more substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO (C)1-C4Alkyl radicals),-CO2H、-CO2(C1-C4Alkyl) and C1-C4An alkoxy group; each of the carboxyl groups mentioned above in the description for formula A, B or C or the variables of formulae 1 to 13 (e.g. R)6、R7、J、R、R'、R"、R*、Ra、RbAnd Rc) Independently is-C (O) O (C)1-C6Alkyl), -OC (O) (C)1-C6Alkyl), -C (O) O (C)3-C6Cycloalkyl), -OC (O) (C)3-C6Cycloalkyl) or-CO2H, wherein the alkyl is optionally substituted with one or more substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO (C)1-C4Alkyl), -CO2H、-CO2(C1-C4Alkyl) and C1-C4An alkoxy group.
Each of the amide groups mentioned in the description for the variables of formula (e.g. formula A, B or C or formulae 1 to 13) disclosed herein is independently-nhc (o) (C)1-C6Alkyl), -N (C)1-C6Alkyl radical C (O) (C)1-C6Alkyl), -C (O) NH (C)1-C6Alkyl), -C (O) N (C)1-C6Alkyl radical)2、-NHC(O)(C3-C6Cycloalkyl), -N (C)1-C6Alkyl radical C (O) (C)3-C6Cycloalkyl), -C (O) NH (C)3-C6Cycloalkyl), -C (O) N (C) 1-C6Alkyl)) (C)3-C6Cycloalkyl) or-C (O) NH2Wherein said alkyl and cycloalkyl are each optionally substituted with one or more substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO (C)1-C4Alkyl), -CO2H、-CO2(C1-C4Alkyl) and C1-C4An alkoxy group.
Each of the aminoalkyl groups mentioned in the description of the variables of formula (e.g., formula A, B or C or formulae 1 to 13) disclosed herein is independently C substituted with one or more amino groups independently selected from the group consisting of1-C6Alkyl groups: -NH2、-NH(C1-C4Alkyl) and-N (C)1-C4Alkyl radical)2。
Each of the aminoalkoxy groups mentioned in the description for the variables of the formulae disclosed herein (e.g., formulae A, B or C or formulae 1 to 13) is independently-O (C)1-C6Alkyl), wherein the alkyl is substituted with one or more amino groups independently selected from the group consisting of: -NH2、-NH(C1-C4Alkyl) and-N (C)1-C4Alkyl radical)2。
In some embodiments, each of the amino groups mentioned in the description for the variables of formula (e.g., formula A, B or C or formulae 1 to 13) disclosed herein is independently-NH2、-NH(C1-C6Alkyl) or-N (C)1-C6Alkyl radical)2Wherein said alkyl groups are each optionally substituted with one or more substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH 2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO (C)1-C4Alkyl), -CO2H、-CO2(C1-C4Alkyl) and C1-C4An alkoxy group.
Each of the carboxyl groups mentioned in the description for the variables of formula (e.g. formula A, B or C or formulae 1 to 13) disclosed herein is independently-C (O) O (C)1-C6Alkyl), -OC (O) (C)1-C6Alkyl) or-CO2H, wherein the alkyl groups are each optionally substituted with one or more substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl radicals),-N(C1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO (C)1-C4Alkyl), -CO2H、-CO2(C1-C4Alkyl) and C1-C4An alkoxy group.
Each of the amide groups mentioned in the description for the variables of formula (e.g. formula A, B or C or formulae 1 to 13) disclosed herein is independently-nhc (o) (C)1-C6Alkyl), -N (C)1-C6Alkyl radical C (O) (C)1-C6Alkyl), -C (O) NH (C)1-C6Alkyl), -C (O) N (C)1-C6Alkyl radical)2or-C (O) NH2Wherein said alkyl groups are each optionally substituted with one or more substituents independently selected from the group consisting of: halogen, cyano, hydroxy, oxo, -NH2、-NH(C1-C4Alkyl), -N (C)1-C4Alkyl radical)2、-OCO(C1-C4Alkyl), -CO (C)1-C4Alkyl), -CO2H、-CO2(C1-C4Alkyl) and C1-C4An alkoxy group.
Each of the aminoalkyl groups mentioned in the description of the variables of formula (e.g., formula A, B or C or formulae 1 to 13) disclosed herein is independently C substituted with one or more amino groups independently selected from the group consisting of 1-C6Alkyl groups: -NH2、-NH(C1-C4Alkyl) and-N (C)1-C4Alkyl radical)2(ii) a And each of the aminoalkoxy groups mentioned in the description for the variables of the formulae disclosed herein (e.g., formulae A, B or C or formulae 1 to 13) is independently-O (C)1-C6Alkyl), wherein the alkyl is substituted with one or more amino groups independently selected from the group consisting of: -NH2、-NH(C1-C4Alkyl) and-N (C)1-C4Alkyl radical)2。
As used herein, the terms "protecting group" and "protecting group" are interchangeable and refer to an agent used to temporarily block one or more desired functional groups in a compound having multiple reactive sites. In certain embodiments, a protecting group has one or more, or exactly all, of the following characteristics: a) selective addition to functional groups in good yield, resulting in b) a protected substrate that is stable to reactions occurring at one or more of the other reactive sites; and c) can be selectively removed in good yield by a reagent that does not attack the regenerated deprotecting functional group. As will be appreciated by those skilled in the art, in some cases, the reagent does not attack other reactive groups in the compound. In other cases, the reagent may also react with other reactive groups in the compound. Examples of protecting Groups are detailed in Greene, T.W., Wuts, P.G, "Protective Groups in Organic Synthesis", third edition, John Wiley & Sons, New York: 1999 (and other versions of the books), the entire contents of which are incorporated herein by reference. As used herein, the term "nitrogen protecting group" refers to an agent used to temporarily block one or more desired nitrogen reactive sites in a polyfunctional compound. Preferred nitrogen protecting groups also have the characteristics exemplified above for the protecting groups, and certain exemplary nitrogen protecting groups are also detailed in Greene, t.w., Wuts, P.G, protecting groups in organic synthesis, third edition, chapter 7, John Wiley & Sons, new york: 1999, the entire contents of which are incorporated herein by reference.
As used herein, the term "displaceable moiety" or "leaving group" refers to a group that is associated with an aliphatic or aromatic group as defined herein and undergoes displacement by nucleophilic attack by a nucleophile.
Unless otherwise indicated, the structures depicted herein are also intended to include all isomeric (e.g., enantiomeric, diastereomeric, cis-trans, conformational and rotational) forms of the structures. For example, unless only one isomer is specifically drawn, the present invention encompasses R and S configurations, (Z) and (E) double bond isomers, and (Z) and (E) configurational isomers of each asymmetric center.
Thus, single stereochemical isomers as well as enantiomeric, diastereomeric, cis/trans, conformational and rotameric mixtures of the compounds of the present invention are within the scope of the invention.
Unless otherwise indicated, all tautomeric forms of the compounds described herein are within the scope of the invention.
In addition, unless otherwise indicated, the structures depicted herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, except for replacement of hydrogen by deuterium or tritium or carbon by 13C or14Compounds having the structure of the present invention other than carbon substitution for C enrichment are within the scope of the present invention. For example, in the reaction with R2Compounds of formulae 1 to 13 having-D at the corresponding position are also within the scope of the invention. Such compounds are useful, for example, as analytical tools or probes in biological assays. These compounds, in particular deuterium analogues, may also have therapeutic applicability.
The terms "a bond" and "absent" are used interchangeably to indicate that a group is absent.
The compounds described herein are defined herein by their chemical structures and/or chemical names. When a compound is referred to by and conflicts with a chemical name through a chemical structure and a chemical name, the chemical structure determines the identity of the compound.
Pharmaceutically acceptable salts
The compounds described herein may be present in free form or, where appropriate, in the form of a salt. Those salts which are pharmaceutically acceptable are of particular interest as they are suitable for administration of the compounds for medical use described below. Non-pharmaceutically acceptable salts are useful in manufacturing processes for isolation and purification purposes, and in some instances, for isolating stereoisomeric forms of the compounds described herein or intermediates thereof.
As used herein, the term "pharmaceutically acceptable salt" refers to salts of compounds which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue side effects (such as toxicity, irritation, allergic response, and the like) commensurate with a reasonable benefit/risk ratio.
Pharmaceutically acceptable salts are well known in the art. For example, s.m. berge et al, in the journal of medical Sciences (j. pharmaceutical Sciences), 1977,66,1-19, which is incorporated herein by reference, describe in detail pharmaceutically acceptable salts. Pharmaceutically acceptable salts of the compounds described herein include those derived from suitable inorganic acids and bases as well as organic acids and bases. These salts can be prepared in situ during the final isolation and purification of the compounds.
In the case where the compounds described herein contain a base or sufficiently basic bioisostere, the acid addition salts may be prepared by 1) reacting the purified compound in its free base form with a suitable organic or inorganic acid and 2) isolating the salt thus formed. In practice, acid addition salts may be in a more suitable form for use, and the use of such salts is equivalent to the use of the free base form.
Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipates, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, formates, fumarates, glucoheptonates, glycerophosphates, glycolates, gluconates, glycolates, hemisulfates, heptanoates, hexanoates, hydrochlorides, hydrobromides, hydroiodides, 2-hydroxy-ethanesulfonates, lactobionates, lactates, laurylsulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoate, embonate, pamoate, and the like, Pectate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate and the like.
In the case where the compounds described herein contain a carboxyl group or a sufficiently acidic bioisostere, base addition salts can be prepared by 1) reacting the purified compound in its acid form with a suitable organic or inorganic base and 2) isolating the salt thus formed. In practice, the use of base addition salts may be more suitable, and the use of the salt form itself corresponds to the use of the free acid form. Salts derived from suitable bases include alkali metals (e.g., sodium, lithium, and potassium), alkaline earth metals (e.g., magnesium and calcium), ammonium, and N+(C1-4Alkyl radical)4And (3) salt. The present disclosure also contemplates the quaternization of any basic nitrogen-containing group of the compounds disclosed herein. Water-or oil-soluble or dispersible products are obtained by such quaternization.
Base addition salts include pharmaceutically acceptable metal and amine salts. Suitable metal salts include sodium, potassium, calcium, barium, zinc, magnesium and aluminum. Sodium and potassium salts are generally preferred. Other pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium and amine cations formed using counter ions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate. Suitable inorganic base addition salts are prepared from metal bases including sodium hydride, sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, zinc hydroxide, and the like. Suitable amine base addition salts are prepared from amines which are commonly used in pharmaceutical chemistry because of their low toxicity and acceptability for medical use. Ammonia, ethylenediamine, N-methyl-glucamine, lysine, arginine, ornithine, choline, N' -benzhydrylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, diethylamine, piperazine, tris (hydroxymethyl) -aminomethane, tetramethylammonium hydroxide, triethylamine, benzhydrylamine, diphenylhydroxymethylamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids, dicyclohexylamine, and the like.
Other acids and bases, when not pharmaceutically acceptable per se, may be used to prepare salts useful as intermediates in obtaining the compounds described herein and pharmaceutically acceptable acid or base addition salts thereof.
It is to be understood that the present disclosure encompasses mixtures/combinations of different pharmaceutically acceptable salts, and also encompasses mixtures/combinations of the compound in free form and the pharmaceutically acceptable salts.
Pharmaceutical composition
The compounds described herein can be formulated into pharmaceutical compositions further comprising a pharmaceutically acceptable carrier, diluent, adjuvant, or vehicle. In some embodiments, the invention relates to a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable carrier, diluent, adjuvant, or vehicle. In some embodiments, the present disclosure encompasses a pharmaceutical composition comprising a safe and effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, adjuvant, or vehicle. Pharmaceutically acceptable carriers include, for example, pharmaceutical diluents, excipients or carriers appropriately selected with respect to the intended form of administration and in accordance with conventional pharmaceutical practice.
An "effective amount" includes both a "therapeutically effective amount" and a "prophylactically effective amount". The term "therapeutically effective amount" refers to an amount effective in treating and/or ameliorating influenza virus infection in a patient. The term "prophylactically effective amount" refers to an amount effective in preventing and/or substantially reducing the chance or size of an outbreak of an influenza virus infection.
Pharmaceutically acceptable carriers can contain inert ingredients that do not unduly inhibit the biological activity of the compound. Pharmaceutically acceptable carriers should be biocompatible, e.g., non-toxic, non-inflammatory, non-immunogenic, or free of other undesirable reactions or side effects when administered to an individual. Standard pharmaceutical compounding techniques may be employed.
A pharmaceutically acceptable carrier, adjuvant, or vehicle, as used herein, includes any solvent, diluent, or other liquid vehicle, dispersing or suspending aid, surfactant, isotonic agent, thickening or emulsifying agent, preservative, solid binder, lubricant, and the like, in a form suitable for the particular dosage form desired. Remington's Pharmaceutical Sciences, 16 th edition, e.w. martin (Mack Publishing co., Easton, Pa.,1980) discloses various carriers for formulating pharmaceutically acceptable compositions and known techniques for their preparation. Unless any conventional carrier medium is incompatible with the compounds described herein, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component of a pharmaceutically acceptable composition, its use is contemplated to be within the scope of the present disclosure. As used herein, the phrase "side effects" encompasses undesirable and adverse effects of a therapy (e.g., prophylactic or therapeutic agent). Side effects are always undesirable, but undesirable as not necessarily undesirable. Adverse effects of therapy (e.g., prophylactic or therapeutic agents) can be harmful or uncomfortable or at risk. Side effects include, but are not limited to, fever, chills, lethargy, gastrointestinal toxicity (including gastric and intestinal ulcers and erosions), nausea, vomiting, neurotoxicity, nephrotoxicity (including conditions including, e.g., papillary necrosis and chronic interstitial nephritis), hepatotoxicity (including elevated serum liver enzyme content), bone marrow toxicity (including leukopenia, myelosuppression, thrombocytopenia, and anemia), xerostomia, metallic taste, prolongation of pregnancy, weakness, somnolence, pain (including muscle pain, bone pain, and headache), hair loss, fatigue, dizziness, extravertebral symptoms, akathisia, cardiovascular disorders, and sexual dysfunction.
Some examples of materials that can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate; lecithin; serum proteins (e.g., human serum albumin); buffer substances (such as twin 80, phosphate, glycine, sorbic acid or potassium sorbate); partial glyceride mixtures of saturated vegetable fatty acids; water, salts or electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride or zinc salts); colloidal silicon dioxide; magnesium trisilicate; polyvinylpyrrolidone; a polyacrylate; a wax; a polyethylene-polyoxypropylene block polymer; methyl cellulose; hydroxypropyl methylcellulose; lanolin; sugars such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered gum tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol or polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; no pyrogen water; isotonic physiological saline; ringer's solution (Ringer's solution); ethanol and phosphate buffer solution; and other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate; as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preserving and anti-oxidants may also be present in the composition according to the judgment of the formulator.
Formulations for pulmonary delivery
In some embodiments, the pharmaceutical compositions described herein are suitable for administration directly via the respiratory tract to the lower respiratory tract (e.g., lungs) by inhalation. Compositions for administration by inhalation may be in the form of inhalable powder compositions or liquid or powder sprays and may be administered in standard form using powder inhalation devices or aerosol dispensing devices. Such devices are well known. For administration by inhalation, powdered formulations typically include the active compound in combination with an inert solid powdered diluent, such as lactose or starch. The inhalable dry powder compositions may be presented in capsules and cartridges of gelatin or similar material or blisters of laminated aluminum foil for use in an inhaler or insufflator. Each capsule or cartridge may typically contain, for example, from about 10mg to about 100g of each active compound. Alternatively, the compositions described herein may be presented without excipients.
The inhalable compositions may be packaged for unit-dose or multi-dose delivery. For example, the composition may be packaged for multiple dose delivery in a manner similar to that described in: GB2242134, U.S. patent nos. 6,632,666, 5,860,419, 5,873,360 and 5,590,645 (all describe "Diskus" devices); or GB2i78965, GB2129691, GB2169265, U.S. patent nos. 4,778,054, 4,811,731 and 5,035,237 (which illustrate "Diskhaler" devices); or EP 69715 ("Turbuhaler" device) or GB 2064336 and us 4,353,656 ("Rotahaler" device).
Spray compositions for delivery to the lung via an inhalation surface may be formulated as aqueous solutions or suspensions or as aerosols delivered from pressurised packs, such as Metered Dose Inhalers (MDI), using a suitable liquefied propellant, comprising hydrofluoroalkanes, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane and especially 1,1,1, 2-tetrafluoroethane, 1,1,1,2,3,3, 3-heptafluoro-n-propane and mixtures thereof. Aerosol compositions suitable for inhalation may be presented in the form of a suspension or solution.
Drugs administered by inhalation typically have a control particle size. The optimum particle size for inhalation into the bronchial system is typically from about 1 μm to about 10 μm, and in some embodiments, from about 2 μm to about 5 μm. Particles having a particle size greater than about 20 μm are generally too large to reach the smaller respiratory tract upon inhalation. To achieve these particle sizes, the particles of the active ingredient may be subjected to a particle size reduction process, such as micron sizing. The desired particle size fraction can be separated by air classification or sieving. Preferably, the particles will be crystalline.
Intranasal sprays may be formulated with aqueous or non-aqueous vehicles with the addition of agents such as thickeners, buffer salts or pH adjusting acids or bases, isotonic adjusting agents or antioxidants.
Solutions for inhalation by nebulization can be formulated using aqueous vehicles with the addition of agents such as acids or bases, buffer salts, isotonic adjusting agents or antimicrobial agents. It may be sterilized by filtration or heating in an autoclave, or presented as a non-sterile product. The atomizer supplies the aerosol in the form of a mist generated from the aqueous formulation.
In some embodiments, the pharmaceutical compositions described herein may be formulated with supplemental active ingredients.
In some embodiments, the pharmaceutical compositions described herein are administered from a dry powder inhaler.
In other embodiments, the pharmaceutical compositions described herein are prepared byThe aerosol dispensing device optionally incorporating e.g. aThe suction chamber of the suction chamber.
The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, liquid polyethylene glycol, and the like), suitable mixtures thereof, and/or vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants. Prevention of the effects of microorganisms in the compositions described herein can be achieved by the addition of antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it is preferred to include isotonic agents, for example sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.
In some embodiments, the pharmaceutical compositions described herein can be within a matrix that controls the release of the composition. In some embodiments, the matrix may comprise: lipids, polyvinyl alcohols, polyvinyl acetates, polycaprolactones, poly (glycolic acid), poly (lactic acid), polycaprolactones, polylactic acids, polyanhydrides, polylactide-co-glycolides, polyamino acids, polyethylene oxides, acrylic acid-capped polyethylene oxides, polyamides, polyethylene, polyacrylonitrile, polyphosphazenes, poly (orthoesters), Sucrose Acetate Isobutyrate (SAIB), and combinations thereof, as well as other polymers disclosed, for example, in U.S. patent nos. 6,667,371, 6,613,355, 6,596,296, 6,413,536, 5,968,543, 4,079,038, 4,093,709, 4,131,648, 4,138,344, 4,180,646, 4,304,767, 4,946,931, each of which is expressly incorporated herein by reference in its entirety. In these embodiments, the matrix provides sustained release of the drug.
The pharmaceutically acceptable carrier and/or diluent may also comprise any solvent, dispersion medium, coating, antibacterial and/or antifungal agent, isotonic and absorption delaying agent, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in pharmaceutical compositions is contemplated.
The pharmaceutical compositions described herein may be formulated for administration according to conventional techniques. See, e.g., Remington, "The Science and Practice of Pharmacy (20 th edition, 2000). For example, intranasal pharmaceutical compositions of the present disclosure may be formulated as aerosols (this term includes both liquid and dry powder aerosols). As known to those skilled in the art, aerosols of liquid particles may be generated by any suitable means, such as using a pressure-driven aerosol atomizer or an ultrasonic atomizer. See, for example, U.S. Pat. No. 4,501,729. Aerosols of solid particles (e.g., lyophilized, freeze-dried, etc.) may likewise be generated by techniques known in the medical arts using any solid particulate medicament aerosol generator. As another example, a pharmaceutical composition of the present disclosure may be formulated in an on-demand soluble form that provides a lyophilized portion of the pharmaceutical composition and a dissolved solution portion of the pharmaceutical composition.
In some embodiments of the invention, the pharmaceutical composition is in the form of an aqueous suspension, which may be prepared from a solution or suspension. For solutions or suspensions, the dosage form may comprise micelles of lipophilic substances, liposomes (phospholipid vesicles/membranes) and/or fatty acids (e.g. palmitic acid). In particular embodiments, the pharmaceutical composition is a solution or suspension capable of dissolving in a fluid secreted by the epithelial cell mucosa of the tissue to which the pharmaceutical composition is applied, applied and/or delivered, which desirably enhances absorption.
The pharmaceutical composition may be an aqueous solution, a non-aqueous solution, or a combination of aqueous and non-aqueous solutions.
Suitable aqueous solutions include (but are not limited to): hydrogels, aqueous suspensions, aqueous microsphere dispersions, aqueous liposome dispersions, aqueous micelles of liposomes, aqueous microemulsions and any combination of the foregoing or any other aqueous solution that is soluble in the fluids secreted by the nasal mucosa. Exemplary non-aqueous solutions include (but are not limited to): non-aqueous gels, non-aqueous suspensions, non-aqueous microsphere dispersions, non-aqueous liposome dispersions, non-aqueous emulsions, non-aqueous microemulsions and any combination of the foregoing or any other non-aqueous solution that is soluble or miscible in the fluid secreted by the mucosa.
Examples of powder formulations include, but are not limited to: pure powder mixtures, micronized powders, freeze-dried powders, lyophilized powders, powder microspheres, coated powder microspheres, liposome dispersions, and any combination of the foregoing. The powder microspheres may be formed from a variety of polysaccharides and celluloses, including, but not limited to, starch, methylcellulose, xanthan gum, carboxymethylcellulose, hydroxypropylcellulose, carbomer, polyvinyl alcohol alginate, acacia, polyglucose, and any combination thereof.
In particular embodiments, the composition is one that is at least partially or even largely (e.g., at least 80%, 90%, 95%, or more) soluble in fluids secreted by the mucosa so as to facilitate absorption. Alternatively or additionally, the composition may be formulated with: carriers and/or other substances that facilitate dissolution of the agent into the secretions include, but are not limited to, fatty acids (e.g., palmitic acid), gangliosides (e.g., GM-1), phospholipids (e.g., phosphatidylserine), and emulsifiers (e.g., polysorbate 80).
Those skilled in the art will appreciate that for intranasal administration or delivery, nasal secretions can alter the pH of the administered dose as the pH range in the nasal cavity can be as broad as 5 to 8, since the volume of pharmaceutical composition administered is generally small. Such changes may affect the concentration of the non-ionized drug available for absorption. Thus, in representative embodiments, the pharmaceutical composition further comprises a buffering agent to maintain or adjust pH in situ. Typical buffers include, but are not limited to, ascorbate, acetate, citrate, gluten, carbonate, and phosphate buffers.
In embodiments of the invention, the pH of the pharmaceutical composition is selected such that the internal environment of the mucosal tissue is acidic to neutral after administration, which (1) can provide the active compound in non-dissociated form for absorption, (2) prevents growth of pathogenic bacteria more likely to be present in an alkaline environment, and (3) reduces the likelihood of mucosal irritation.
For liquid and powder sprays or aerosols, the pharmaceutical compositions may be formulated to have any suitable and desired particle size or droplet size. In illustrative embodiments, the majority and/or average particle size of the particles or droplets ranges from equal to or greater than about 1, 2.5, 5, 10, 15, or 20 microns and/or equal to or less than about 25, 30, 40, 45, 50, 60, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, or 425 microns (including all combinations of the foregoing). Representative examples of suitable ranges for the majority and/or average particle size or droplet size include, but are not limited to, about 5 to 100 microns, about 10 to 60 microns, about 175 to 325 microns, and about 220 to 300 microns, which facilitate deposition of a safe and effective amount of the active compound, for example, in the nasal cavity (e.g., in the upper one-third of the nasal cavity, the upper nasal passage, the olfactory region, and/or the sinus region leading to the olfactory nerve pathway). Generally, particles or droplets smaller than about 5 microns will deposit in the trachea or even the lungs, while particles or droplets of about 50 microns or larger generally do not reach the nasal cavity and deposit on the anterior portion of the nose.
International patent publication WO 2005/023335(Kurve Technology, Inc.) describes particles and droplets having a diameter size suitable for practicing representative embodiments of the present disclosure. In particular embodiments, the particles or droplets have an average diameter of about 5 to 30 microns, about 10 to 20 microns, about 10 to 17 microns, about 10 to 15 microns, about 12 to 17 microns, about 10 to 15 microns, or about 10 to 12 microns. The particles may be "substantially" of an average diameter or size as described herein, i.e., at least about 50%, 60%, 70%, 80%, 90%, or 95% or more of the particles have the indicated diameter or size range.
The pharmaceutical compositions described herein may be delivered in the form of a nebulized or nebulized liquid having a droplet size as described above.
According to particular embodiments of the present disclosure that include intranasal delivery methods, it may be desirable to prolong the residence time of the pharmaceutical composition in the nasal cavity (e.g., in the upper third of the nasal cavity, the upper nasal passage, the olfactory region, and/or the sinus region), e.g., to enhance absorption. Thus, the pharmaceutical composition may optionally be formulated with: bioadhesive polymers, gums (e.g. xanthan gum), polyglucose (e.g. highly purified cationic polysaccharides), pectin (or any carbohydrate that thickens or emulsifies like a gel when applied to the nasal mucosa), microspheres (e.g. starch, albumin, polydextrose, cyclodextrin), gelatin, liposomes, carbomers, polyvinyl alcohol, alginates, acacia, polyglucose and/or cellulose (e.g. methyl or propyl cellulose; hydroxy or carboxy cellulose; carboxymethyl or hydroxypropyl cellulose), which are agents that increase the residence time in the nasal cavity. As another approach, increasing the viscosity of the formulation may also provide a means of prolonging contact of the agent with the nasal epithelial cells. The pharmaceutical composition may be formulated as a nasal emulsion, ointment or gel, which provides the advantage of topical administration due to its viscosity.
Moist and highly vascularized membranes can contribute to rapid absorption; thus, the pharmaceutical composition may optionally comprise a humectant, especially in the case of gel-based compositions, to ensure sufficient intranasal moisture content. Examples of suitable humectants include, but are not limited to, glycerol (glycerin/glycerol), mineral oil, vegetable oil, membrane regulators, soothing agents, and/or sugar alcohols (e.g., xylitol, sorbitol; and/or mannitol). The concentration of the humectant in the pharmaceutical composition will vary depending on the agent and formulation selected.
The pharmaceutical composition may also optionally comprise absorption enhancers, such as agents that inhibit enzyme activity, reduce viscosity or elasticity of mucus, reduce mucociliary clearance effects, open tight junctions, and/or solubilize the active compound. Chemical enhancers are known in the art and include chelating agents (e.g., EDTA), fatty acids, cholates, surfactants, and/or preservatives. Enhancers for permeation may be particularly useful when formulating compounds that exhibit poor membrane permeability, lack lipophilicity, and/or are degraded by aminopeptidases. The concentration of the absorption enhancer in the pharmaceutical composition will vary depending on the agent selected and the formulation.
Preservatives may optionally be added to the pharmaceutical composition in order to prolong shelf life. Suitable preservatives include, but are not limited to, benzyl alcohol, parabens, thimerosal, chlorobutanol, and benzalkonium chloride, and combinations of the foregoing. The concentration of the preservative will vary depending on the preservative used, the compound formulated, the formulation, and the like. In representative embodiments, the preservative is present in an amount of about 2% by weight or less.
The pharmaceutical compositions described herein may optionally contain an odorant, for example as described in EP 0504263B 1, to provide a sensation of odor in order to facilitate inhalation of the composition, thereby facilitating delivery to the olfactory region and/or triggering transmission by olfactory neurons.
As another option, the composition may include a flavoring agent, for example, to enhance taste and/or individual acceptability of the composition.
Porous particles for pulmonary administration
In some embodiments, the particles are porous such that they have a suitable density to avoid deposition in the back of the throat when administered via an inhaler. The combination of relatively large particle size and relatively low density avoids phagocytosis in the lung, provides for properly targeted delivery, avoids systemic delivery of components and provides for high concentrations of components within the lung.
Representative methods for preparing such particles and for delivering such particles are described in the following: for example, U.S. patent No. 7,384,649 entitled "particle compositions for pulmonary delivery"; U.S. patent No. 7,182,961 entitled "particle compositions for pulmonary delivery"; U.S. patent No. 7,146,978 entitled "Inhalation device and method"; U.S. patent No. 7,048,908 entitled "Particles for inhalation having sustained release properties"; U.S. patent No. 6,956,021 entitled "Stable spray-dried protein formulations"; U.S. patent No. 6,766,799 entitled "Inhalation device"; and U.S. patent No. 6,732,732 entitled Inhalation device and method.
Additional patents disclosing such particles include: U.S. patent No. 7,279,182 entitled "Formulation for spray-drying porous large particles"; U.S. Pat. No. 7,252,840 entitled "formation of porous particles using pure amino acids (Use of simple amino acids to form porous particles)"; U.S. patent No. 7,032,593 entitled "Inhalation device and method"; U.S. patent No. 7,008,644 entitled "Method and apparatus for producing dried particles"; U.S. Pat. No. 6,848,197 entitled "Control of Process humidity to Process Large, porous particles"; and U.S. patent No. 6,749,835 entitled "Formulation for spray-drying porous large particles".
U.S. patent No. 7,678,364, entitled "Particles for inhalation having sustained release properties," discloses a method for delivering Particles to the pulmonary system comprising: administering to the respiratory tract of a patient in need of treatment, prevention, or diagnosis a safe and effective amount of a dry powder comprising: a) a polyvalent metal cation complexed with a therapeutic, prophylactic or diagnostic agent, b) a pharmaceutically acceptable carrier, and c) a polyvalent metal cation-containing component, wherein the dry powder is spray-dried and has a total amount of polyvalent metal cations of about 10% w/w or more, about 0.4g/cm, based on the total weight of the medicament3Or less, a tap density of about 5 microns to about 30 microns, a median geometric diameter, and an aerodynamic diameter of about 1 to about 5 microns.
The amount of a compound described herein or salt thereof present in the particles can range from about 0.1 wt% to about 95 wt%, although in some cases, can even be as high as 100%. For example, from about 1 to about 50%, such as from about 5 to about 30%. Particles in which the drug is distributed throughout the particle may be preferred.
In some embodiments, the particles comprise a surfactant other than the phospholipids described above. As used herein, the term "surfactant" refers to any agent that preferentially absorbs to the interface between two immiscible phases (e.g., the interface between water and an organic polymer solution, the water/air interface, or the organic solvent/air interface). Surfactants generally have a hydrophilic portion and a lipophilic portion such that, upon absorption into the particle, they tend to bring the portions to an external environment that does not attract particles like the coating, thus reducing particle aggregation. Surfactants may also facilitate the absorption of therapeutic or diagnostic agents and increase the bioavailability of the agents.
Suitable surfactants that may be used in the manufacture of the particles described herein include (but are not limited to): cetyl alcohol; fatty alcohols, such as polyethylene glycol (PEG); polyoxyethylene-9-lauryl ether; surface active fatty acids, such as palmitic acid or oleic acid; glycocholate; surface active peptides (surfactin); poloxamers (poloxamers); sorbitan fatty acid esters, such as sorbitan trioleate (Span 85);80; and tyloxapol (tyloxapol).
The surfactant may be present in the particles in an amount in the range of about 0 to about 5 weight percent. Preferably, it may be present in the particles in an amount in the range of about 0.1 to about 1.0 wt%.
Having a density of less than about 0.4g/cm3Particles of tap density of at least about 5 μm, median diameter of about 1 μm to about 5 μm, or aerodynamic diameter of about 1 μm to about 3 μm are more capable of avoiding inertial and gravitational deposition in the oropharyngeal region and are targeted to the respiratory tract or deep lung. The use of larger, more porous particles is advantageous because it enables more efficient aerosolization than smaller, denser aerosol particles (such as those currently used for inhalation therapy).
Liposome delivery
The compositions described herein are preferably delivered to the lung so as to provide the compound at the site of actual or potential influenza infection. This can be achieved by pulmonary delivery via a metered dose inhaler or other pulmonary delivery device, and also by passing the particles into the microvascular bed surrounding the alveoli in the lung.
Nanocarriers comprising smaller unilamellar vesicles (such as liposomes) exhibit several advantages over other conventional methods for drug delivery to the lung, including prolonged drug release and cell-specific targeted drug delivery. Nanoscale drug carriers may also be advantageous for delivery of drugs that are poorly water soluble, and some of the compounds described herein are poorly water soluble. Additional advantages include its ability to provide controlled release, protection from metabolism and degradation, reduced drug toxicity and targeting ability.
Liposomes (preferably unilamellar vesicles) have a size of less than 200nm as measured by dynamic light scattering and are preferably characterized by comprising a chemically pure synthetic phospholipid composition, most preferably having an aliphatic side chain of at least 16 carbons in length, and containing one or more of the compounds described herein or a pharmaceutically acceptable salt thereof sufficient to deliver (i.e., target) an amount of its compound preferably to the microvascular bed surrounding the alveoli. The vesicle diameter can be measured, for example, by dynamic light scattering using a helium-neon 100mW NEC gas laser and a Malvern K7027 correlator, ideally producing at least two or three measurements at a time for each size determination.
The expression "chemically pure phospholipids" is intended to define phospholipids substantially free of harmful decontaminating moieties and impurities causing the polymerization of Smaller Unilamellar Vesicles (SUV) formed therefrom and having a purity of more than 97%. Preferably, the liposomes are predominantly about 50 to about 160nm in diameter, substantially neutral in charge, and incorporate phospholipids having side chains of 16 to 18 carbon atoms in length. More preferably, the liposomes are prepared from Distearoylphosphatidylcholine (DSPC) and contain cholesterol as a vesicle stabilizer (most preferably, in an amount of 10% to 50% of the total lipids).
It may also be advantageous for the liposomes to have a melting point above body temperature (i.e., greater than 37 ℃). Thus, it may be advantageous to use pure phospholipids, preferably saturated phospholipids, having a carbon chain length of at least 16 carbons, preferably between 16 and 18 carbons. Distearoylphosphatidylcholine (DSPC) is a preferred phospholipid. Cholesterol helps stabilize the liposomes and is preferably added in an amount sufficient to provide liposome stability. Most preferably, the liposome further comprises a pegylated phospholipid, such as dspeg. The methods involve introducing into the bloodstream of a patient an amount of liposomes that are less than 200nm in size (preferably unilamellar vesicles) and are preferably characterized by comprising a chemically pure synthetic phospholipid, most preferably having an aliphatic side chain of at least 16 carbons in length, and containing a compound described herein or a pharmaceutically acceptable salt or prodrug thereof sufficient to deliver (i.e., target) an amount of the compound, preferably to the microvascular bed surrounding the alveoli in the lung.
The compounds described herein may be combined with other anti-influenza agents, also as described herein. Such additional agents may also be present in the liposomes, may be present in different liposomes, or may be co-administered via different routes.
The liposomes comprise one or more of the compounds described herein, or a pharmaceutically acceptable salt thereof, and may optionally comprise other anti-influenza agents. The liposome can be prepared by dissolving phospholipid and cholesterol in a suitable organic solvent such as chloroform and evaporating the solvent to form a lipid film. If an ionophore is used to load the compounds described herein into liposomes, the ionophore may be added to the lipid solution prior to evaporation. The dried lipid film is then rehydrated in an appropriate aqueous phase, such as phosphate buffered saline or other physiologically suitable solution. The water soluble drug or therapeutic agent may be contained in the hydration solution, but if remote loading is desired, a loading agent such as a chelating agent as described above may be added to the hydration solution to be encapsulated within the internal aqueous space of the liposome.
Upon addition of the hydration solution, liposomes of different sizes spontaneously form and encapsulate a portion of the aqueous phase. Thereafter, the liposomes and the aqueous suspension solution are subjected to a shearing force such as extrusion, sonic treatment or treatment via a homogenizer according to the method described in U.S. Pat. No. 4,753,788; to produce vesicles within a specified size.
The liposomes can then be treated to remove undesirable compounds, such as unencapsulated drugs, from the suspension solution, which can be achieved via processes such as gel chromatography or ultrafiltration.
The use of liposomes in dry powder aerosols for targeted Lung delivery is described, for example, in Willis et al, lungs (Lung), 6.2012, 190(3): 251-262. One advantage is that the phospholipids used to prepare the liposomes are similar to endogenous lung surfactants.
Application method
Depending on the severity of the infection being treated, the compounds and pharmaceutically acceptable compositions described above may be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (as powders, ointments, or drops), buccally, as an oral or nasal spray, to the pulmonary system (e.g., by using an inhaler, such as a Metered Dose Inhaler (MDI)), or the like.
Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, liquid dosage forms may contain: inert diluents commonly used in the art, such as water or other solvents; co-solvents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, dimethylformamide, oils (in detail, cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan; and mixtures thereof. In addition to inert diluents, oral compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
For example, injectable preparations, such as sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S. P, and isotonic sodium chloride solution. In addition, sterile, nonvolatile oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed, including synthetic mono-or diglycerides. In addition, fatty acids (such as oleic acid) are used in the preparation of injectables.
The injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable medium at the point of use.
To prolong the effect of the compounds described herein, it is generally desirable to slow the absorption of the compounds from subcutaneous or intramuscular injection. This can be achieved by using liquid suspensions of crystalline or amorphous materials with poor water solubility. The rate of absorption of the compound depends on its rate of dissolution, which in turn may depend on crystal size and crystalline form. Alternatively, delayed absorption of a compound for parenteral administration is achieved by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming a microencapsulated matrix of the compound in a biodegradable polymer such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the nature of the particular polymer used, the rate of release of the compound can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Injectable depot formulations can also be prepared by encapsulating the compounds in liposomes or microemulsions that are compatible with body tissues.
Compositions for rectal or vaginal administration are in particular suppositories which can be prepared by mixing the compounds described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with: at least one inert, pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate; and/or a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders such as carboxymethyl cellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and acacia; c) humectants, such as glycerol; d) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; e) solution retarding agents, such as paraffin; f) absorption promoters, such as quaternary ammonium compounds; g) wetting agents, such as cetyl alcohol and glycerol monostearate; h) absorbents such as kaolin and bentonite clay; and i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.
Solid compositions of a similar type may also be employed as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose (lactose/milk sugar) and high molecular weight polyethylene glycols and the like. Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art of pharmaceutical formulation. It may optionally contain opacifying agents and may also have a composition that releases the active ingredient only or preferably in a certain part of the intestinal tract, or optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose (lactose/milk sugar) and high molecular weight polyethylene glycols and the like.
The active compound may also be in microencapsulated form with one or more excipients as described above. Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release-controlling coatings, and other coatings well known in the art of pharmaceutical formulation. In such solid dosage forms, the active compound may be mixed with at least one inert diluent (e.g., sucrose, lactose or starch). As in general practice, such dosage forms may also include other substances in addition to inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. It may optionally contain opacifying agents and may also have a composition that releases the active ingredient only or preferably in a certain part of the intestinal tract, or optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.
Dosage forms for topical or transdermal administration of the compounds described herein include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier and any required preservatives or buffers as may be desired. Ophthalmic formulations, ear drops, and eye drops are also contemplated as falling within the scope of the present disclosure. In addition, the present disclosure contemplates the use of transdermal patches, which have the additional advantage of providing controlled delivery of compounds to the body. Such dosage forms may be prepared by dissolving or dispensing the compound in the appropriate medium. Absorption enhancers may also be used to increase the flux of the compound through the skin. The rate can be controlled by providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.
The compositions described herein may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implantable reservoir. As used herein, the term "parenteral" includes, but is not limited to, subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. In particular, the compositions are administered orally, intraperitoneally, or intravenously.
The sterile injectable form of the compositions described herein can be an aqueous or oleaginous suspension. These suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution. In addition, sterile, nonvolatile oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. Fatty acids (such as oleic acid and its glyceride derivatives) are suitable for the preparation of injectables, as are natural pharmaceutically-acceptable oils (such as olive oil or castor oil, especially in their polyoxyethylated versions). These oil solutions or suspensions may also contain a long chain alcohol diluent or dispersant, such as carboxymethyl cellulose or similar dispersing agents commonly used in formulating pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants (such as Tween, Span, and other emulsifiers) or bioavailability enhancers commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for formulation purposes.
The pharmaceutical compositions described herein may be administered orally in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, commonly used carriers include, but are not limited to, lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in capsule form, suitable diluents include lactose and dried corn starch. When aqueous for oral use is desired, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.
Alternatively, the pharmaceutical compositions described herein may be administered in the form of suppositories for rectal administration. These suppositories can be prepared by mixing the agent with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include, but are not limited to, cocoa butter, beeswax and polyethylene glycols.
The pharmaceutical compositions described herein may also be administered topically, particularly when the target of treatment comprises topical administration of an easily accessible area or organ, including diseases of the eye, skin or lower intestinal tract. Suitable surface formulations for each of these regions or organs are readily prepared.
Topical administration for the lower intestinal tract may be achieved in the form of rectal suppository formulations (see above) or in the form of suitable enema formulations. Topical transdermal patches may also be used.
For topical administration, the pharmaceutical compositions may be formulated in a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds described herein include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide compounds, emulsifying wax, and water. Alternatively, the pharmaceutical compositions may be formulated in a suitable lotion or cream containing the active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
For ophthalmic use, the pharmaceutical composition may be formulated as a micronized suspension in isotonic pH adjusted sterile saline, or rather as a solution in isotonic pH adjusted sterile saline, with or without a preservative (such as benzalkonium chloride). Alternatively, for ocular administration, the pharmaceutical composition may be formulated in an ointment such as petrolatum.
The compounds for use in the methods described herein may be formulated in unit dosage form. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for the individual undergoing treatment, wherein each unit contains a predetermined quantity of active material calculated to produce the desired therapeutic effect, optionally in association with a suitable pharmaceutical carrier. The unit dosage form can be used in a single daily dose or in one of a plurality of daily doses (e.g., about 1 to 4 or more times per day). When multiple daily doses are used, the unit dosage forms for each dose may be the same or different.
The present disclosure may be more completely understood in consideration of the examples described herein in detail of exemplary embodiments. However, these examples should not be construed as limiting the scope of the disclosure. All references to the entire disclosure are expressly incorporated herein by reference.
Detailed Description
Examples of the invention
Example 1: side chain preparation
Side chain-1
2- (2- (2-ethoxyethoxy) ethoxy) acetic acid (side chain-1)
To a stirred solution of 2- (2-ethoxyethoxyethoxy-ethan-1-ol 1(2.68g, 20mmol) in 1, 4-dioxane (60mL) at 0 ℃ NaOH (1.2g, 30mmol) was added the mixture was stirred at room temperature for 15 minutes then cooled to 0 ℃, tert-butyl 2-bromoacetate 2(7.8g, 40mmol) and 18-crown ether (250mg) were added to the reaction mixture and stirred at room temperature for 16 hours after the starting material was exhausted, the mixture was diluted with ice-cold water (200mL), extracted with ether (2X 100mL), the separated aqueous layer was acidified (pH about 2) with concentrated HCl (30mL), and extracted with dichloromethane (2 × 250mL) brine solution (50mL), dried over sodium sulfate and concentrated under reduced pressure to give pure 2- (2- (2-ethoxyethoxy) ethoxy) acetic acid as a brown liquid.
Side chain-1 (1.5g, 7.8mmol, 39% yield). TLC system: 10% methanol in dichloromethane: rf：0.10
Side chain-2
4-Methylbenzenesulfonic acid 2- (2- (2-ethoxyethoxy) ethoxy) ethyl ester (2)
To a stirred solution of 2- (2- (2-ethoxyethoxy) ethoxy) ethan-1-ol 1(5.0g, 28.05mmol) in THF (20mL) at 0 deg.C was added a solution of NaOH (2.28g, 57.22mmol) in water (10mL) followed by p-toluenesulfonyl chloride (6.84g, 35.90) at 0 deg.Cmmol) in THF (10 mL). The mixture was then allowed to reach room temperature and stirred for 4 hours. After the starting material was consumed, the mixture was quenched with water (100mL) and extracted with ether (2 × 100mL), washed with ice-cold water (2 × 25mL), brine solution (25mL), dried over sodium sulfate, and concentrated to give 2- (2- (2-ethoxyethoxy) ethoxy) ethyl 4-methylbenzenesulfonate 2(9.02g, 27.16mmol, 96% yield) as a colorless oil. TLC system: 40% ethyl acetate in hexane, Rf：0.3；LCMS：m/z＝332.83(M+H)+
2- (2- (2- (2-ethoxyethoxy) ethoxy) ethyl) isoindoline-1, 3-dione (3)
To a stirred solution of 4-methylbenzenesulfonic acid 2- (2- (2-ethoxyethoxy) ethoxy) ethyl ester 2(6g, 18.07mmol) in DMF (15mL) at room temperature was added potassium phthalimide (4.418g, 23.85mmol), followed by stirring at 110 ℃ for 16 hours. After completion of the reaction as indicated by TLC, the reaction mixture was allowed to cool to room temperature, diethyl ether (50mL) was added and stirred at room temperature for 15 minutes, filtered and washed with diethyl ether. The filtrate was washed with 1M NaOH solution (50mL), water (50mL), brine solution (50mL), and Na 2SO4Dried and concentrated under reduced pressure to give 2- (2- (2- (2-ethoxyethoxy) ethoxy) ethyl) isoindoline-1, 3-dione 3(3.5g, 11.40mmol, 63%) as a yellow oily liquid. TLC system: 40% ethyl acetate in hexane, Rf：0.50；LCMS：m/z＝308.11(M+H)+
2- (2- (2-ethoxyethoxy) ethoxy) ethan-1-amine (side chain-2)
To a stirred solution of 2- (2- (2- (2-ethoxyethoxy) ethoxy) ethyl) isoindoline-1, 3-dione 3(500mg, 1.62mmol) in ethanol (5mL) was added hydrazine hydrate (5mL) and stirred at 110 ℃ for 16 h. Such asAfter TLC indicated completion of the reaction, the reaction mixture was cooled to room temperature and extracted with toluene (2 × 50mL) and washed with brine solution (25mL), Na2SO4Dried and concentrated under reduced pressure to give 2- (2- (2-ethoxyethoxy) ethoxy) ethan-1-amine 4(150mg, 0.84mmol, 52%) as a yellow oily liquid. TLC system: 10% methanol in dichloromethane, Rf：0.10
Side chain-3
2- (2- (2-ethoxyethoxy) ethoxy) -N-methylethyl-1-amine (side chain-3)
A stirred solution of 4-methylbenzenesulfonic acid 2- (2- (2-ethoxyethoxy) ethoxy) ethyl ester 1 (the synthesis of compound 1 is reported in side chain-2) (500mg, 1.50mmol) and 33% methylamine in ethanol solution (3mL) was heated in a sealed tube at 50 ℃ for 16 h. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, the crude material was dissolved in water (25mL) and extracted with dichloromethane (2 × 50 mL). The combined organic layers were washed with brine solution (25mL) and Na 2SO4Drying and concentration gave 2- (2- (2-ethoxyethoxy) ethoxy) -N-methylethyl-1-amine side chain-3 (210mg, 1.09mmol, 73%) as a yellow oily liquid. TLC system: 10% methanol in dichloromethane, Rf：0.20
Side chain-4
2- (2- (2-ethoxyethoxy) ethoxy) ethane-1-thiol (2)
To 4-methylbenzenesulfonic acid 2- (2- (2-ethoxyethoxy) ethoxy) ethyl ester 1 (the synthesis of compound 1 has been reported in side chain-2) (2.2g, 6.626mmol) inTo a stirred solution in ethanol (10mL), THF (1mL) was added sodium hydrogensulfite hydrate (3.7g, 66.26mmol) and stirred at room temperature for 16 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, the crude material was diluted with water (100mL) and extracted into ethyl acetate (2 × 100 mL). The organic layer was washed with brine solution (100mL) over Na2SO4Dried and concentrated to give 2- (2- (2-ethoxyethoxy) ethoxy) ethane-1-thiol 2(1.1g, 5.67mmol, 85%) as a light brown liquid. TLC system: 50% ethyl acetate in hexane; rf:0.20
2- (2- (2-ethoxyethoxy) ethoxy) ethane-1-sulfonyl chloride (side chain-4)
To a stirred solution of 2- (2- (2-ethoxyethoxy) ethoxy) ethane-1-thiol 2(1.4g, 7.21mmol) and potassium nitrate (2.41g, 18.04mmol) was added sulfuryl chloride (1.5ml) at 0 ℃ and stirred at room temperature for 16 hours. After completion of the reaction as indicated by TLC, water (200mL) was added and extracted with DCM (2 × 200mL), the organic layer was washed with brine solution (100mL), and Na was added 2SO4Drying and concentration gave 2- (2- (2-ethoxyethoxy) ethoxy) ethane-1-sulfonyl chloride side chain-4 (1.4g, 5.38mmol, 74%) as a brown liquid. TLC system: 50% ethyl acetate in hexane; rf：0.40
Side chain-5
2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) ethanol (2)
Stirring of 2,2' - (ethane-1, 2-diylbis (oxy)) diethanol 1(50g, 333.3mmol) in dichloromethane (1.5L) at 0 deg.CTo the mixed solution were added dihydropyran (20g, 233.3mmol) and p-toluenesulfonic acid (6.3g, 33.33mmol), and the mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water (1000mL) and extracted with dichloromethane (3 × 800mL), the combined organic layers were washed with brine (2 × 200mL) and dried over sodium sulfate, and concentrated. The residue was purified by flash column chromatography (100-200 silica) using 3% methanol in dichloromethane to give 2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) ethanol 2(3g, 12.82mmol, 10% yield) as a colorless oily liquid. TLC system: 15% acetone in dichloromethane, Rf：0.35
Ethyl 2- (2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) acetate (4)
To a stirred solution of 2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) ethanol 2(2.7g, 11.54mmol) in N, N-dimethylformamide (5mL) at 0 ℃ was added NaH (554mg, 23.08mmol) and stirred at room temperature for 30 min. A solution of ethyl 2-bromoacetate 3(2.3g, 13.84mmol) in N, N-dimethylformamide (5mL) was then added to the above reaction mixture at 0 deg.C and stirred at room temperature under a nitrogen atmosphere for 16 hours. The reaction mixture was quenched with ice-cold water and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine (2X 100mL) and Na 2SO4Drying and evaporating under reduced pressure. The crude residue was purified by combined flash chromatography using 40% ethyl acetate in hexanes to give ethyl 2- (2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) acetate 4(550mg, 1.7187mmol, 15% yield) as a light yellow oily liquid. TLC system: 70% ethyl acetate in hexane; rf：0.50
2- (2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) acetic acid (side chain-5)
To ethyl 2- (2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) acetate 3(550mg, 1.7187mmol) in THF H at 0 deg.C2To a stirred solution of a mixture of O (3:1) (12mL) was added lithium hydroxide monohydrate (288mg, 6.875mmol) and stirred at room temperature for 16 h. The reaction mixture was concentrated to remove organic volatiles, water (50mL) was added to the crude and extracted with ethyl acetate (2 × 20 mL). The aqueous layer was acidified with saturated citric acid solution and extracted with 10% methanol in dichloromethane (2 × 50 mL). The combined organic layers were washed with brine solution (15mL), dried over sodium sulfate, and concentrated to give 2- (2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) acetic acid as a pale yellow gummy liquid (340mg, 1.1643mmol, 68%). TLC system: 10% methanol in dichloromethane; r f：0.10
Side chain-6
4-Methylbenzenesulfonic acid 2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl ester (2)
To a stirred solution of 2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethan-1-ol 1(9.5g, 40.598mmol, (synthesis of compound 1 is reported in side chain-5)) in THF (100mL) at 0 ℃ were added NaOH (3.25g, 81.196mmol) and p-toluenesulfonyl chloride (9.3g, 48.718mmol), followed by stirring at room temperature for 16H. After the starting material was consumed, the mixture was diluted with ethyl acetate (50mL) and washed with ice-cold water (2 × 20mL), brine solution (20mL), dried over sodium sulfate and concentrated under reduced pressure to give 4-methylbenzenesulfonic acid 2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl ester 2 as a colorless oily liquid (14g, 36.08mmol, crude material). TLC system: 70% ethyl acetate in Petroleum Ether, Rf：0.50；LCMS：m/z＝305.04(M-THP)+
2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl) isoindoline-1, 3-dione
To a stirred solution of 4-methylbenzenesulfonic acid 2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl ester 2(14g, 36.08mmol) in DMF (35mL) was added potassium phthalimide (8.8g, 47.62mmol) at room temperature, followed by stirring at 110 ℃ for 16 hours. After completion of the reaction as indicated by TLC, the reaction mixture was concentrated, the residue was suspended in ether and stirred for 15 min and filtered, the filtrate was washed with 1M NaOH solution (2 × 100mL), water (100mL) and brine solution (100mL), dried over sodium sulfate and concentrated under reduced pressure to give 2- (2- (2- (2- (((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl) isoindoline-1, 3-dione 3(9g, 24.79mmol, 69% yield) as a yellow oily liquid a TLC system 40% ethyl acetate in petroleum ether, R, g, R f：0.50；LCMS：m/z＝364(M-THP)+
2- (2- (2- ((tetrahydro-2H-pyran-2-yloxy) ethoxy) ethan-1-amine (4)
To a stirred solution of 2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl) isoindoline-1, 3-dione 3(9g, 24.79mmol) in ethanol was added hydrazine hydrate (18mL) and stirred at 110 ℃ for 16H. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, water (100mL) was added to the crude material and extracted with toluene (3 × 200 mL). The organic layer was washed with brine solution (100mL) over Na2SO4Drying and concentration gave 2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethan-1-amine 4(3.4g, 14.59mmol, 59%)). TLC system: 15% acetone in dichloromethane, Rf：0.10；LCMS：m/z＝234(M+H)+
(2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl) carbamic acid tert-butyl ester (5)
To a stirred solution of 2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethan-1-amine (4) (3.2g, 13.7mmol) in THF (30mL) was added (Boc)2O (3.15mL, 13.73mmol) and 0.1 equivalent of DMAP (167mg, 1.37mmol), followed by stirring at room temperature for 2 hours. After completion of the reaction as indicated by TLC, water (100mL) was added to the reaction mixture and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with brine solution (100mL) and Na 2SO4Dried and concentrated. The crude compound was purified by 100-pack 200 silica gel column chromatography eluting with 25% ethyl acetate in petroleum ether to give tert-butyl (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl) carbamate 5(2.8g, 8.408mmol, 61%) as a colorless liquid. TLC system: 40% ethyl acetate in petroleum ether, Rf：0.50
(2- (2- (2-hydroxyethoxy) ethoxy) ethyl) carbamic acid tert-butyl ester (6)
To a stirred solution of tert-butyl (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl) carbamate 5(2.8g, 8.408mmol) in methanol (30mL) at 0 ℃ was added pyritinol p-toluenesulfonate (1.4g, 5.885mmol) and stirred at room temperature for 16H. The organic solvent was distilled off, water (100mL) was added to the crude material, and extracted with ethyl acetate (3X 150 mL). The combined organic layers were washed with brine solution (100mL), dried over sodium sulfate and concentrated under reduced pressure. Crude Compound 50% in Petroleum Ether by flash column chromatography (100-200 silica)Purification of ethyl acetate gave tert-butyl (2- (2- (2-hydroxyethoxy) ethoxy) ethyl) carbamate 6(1.8g, 7.229mmol, 86% yield) as a colorless gummy liquid. TLC system: 40% ethyl acetate in petroleum ether, R f：0.10
2, 2-dimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azahexadecane-16-oic acid ethyl ester (8)
A stirred solution of tert-butyl (2- (2- (2-hydroxyethoxy) ethoxy) ethyl) carbamate 6(125mg, 0.502mmol) in N, N-dimethylformamide (2mL) was treated with NaH (30mg, 1.255mmol) at 0 deg.C and stirred at room temperature for 30 min. Ethyl 2-bromoacetate 7(125mg, 0.7530mmol) in N, N-dimethylformamide (1mL) was added to the above reaction mixture at 0 ℃ and the mixture was stirred at room temperature under an argon atmosphere for 16 hours. The reaction mixture was quenched with ice water (50mL) and extracted with ethyl acetate (3X 20 mL). The combined organic layers were washed with brine solution (20mL) and Na2SO4Dried and evaporated under reduced pressure. The crude residue was purified by combined flash chromatography using 70% ethyl acetate in petroleum ether to give ethyl 2, 2-dimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azahexadecane-16-carboxylate 8(35mg, 0.144mmol, 20% yield) as a colorless liquid. TLC system: 70% ethyl acetate in petroleum ether, Rf: 0.20; direct quality: 236.1(M-Boc)+
2, 2-dimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azahexadecane-16-oic acid (side chain-6)
To ethyl 2, 2-dimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azahexadecane-16-oate 8(300mg, 0.895mmol) in THF H at room temperature 2To a stirred solution of O (4:1) (20mL) was added lithium hydroxide (112mg, 2.685mmol) for 16 hours. The reaction mixture was concentrated to remove organic volatiles, water (50mL) was added to the residue and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with brine solution (20mL), dried over sodium sulfate and concentrated under reduced pressure to give crude compound 2, 2-dimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azahexadecane-16-carboxylic acid side chain-6 (280mg, 0.9120mmol, crude material) as a colorless liquid. TLC system: 70% ethyl acetate in petroleum ether, Rf: 0.10; direct quality: m/z 208.07(M-Boc)+
Side chain-7
2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) ethanol (2)
To a stirred solution of 2,2' -oxodiethanol 1(25g, 235.58mmol) in dichloromethane (750mL) at 0 deg.C were added dihydropyran (13.8g, 164.85mmol) and pyritinol p-toluenesulfonate (4.48g, 23.55mmol), and the mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water (600mL), extracted into dichloromethane (3 × 250mL), the combined organic layers were washed with brine (2 × 100mL), and dried over sodium sulfate and concentrated. The residue was purified by column chromatography (100-200 silica) using 2% methanol/dichloromethane to give 2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) ethanol 2(7.8g, 41.0mmol, 17% yield) as a pale yellow oily liquid. TLC system: 10% methanol in dichloromethane, R f：0.3
3- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) propionic acid ethyl ester (4)
To NaH (1.74g, 43.42 mmo) at 0 deg.Cl) to a stirred suspension in THF (70mL) was added a solution of 2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) ethanol 2(5.5g, 28.947mmol) in THF (20mL) and stirred at room temperature for 30 min. Ethyl 3-bromopropionate 3(7.85g, 43.42mmol) in THF (10mL) was added to the above reaction mixture at 0 ℃ and stirred at room temperature under a nitrogen atmosphere for 16 hours. The reaction mixture was quenched with ice water (200mL) and extracted with ethyl acetate (3X 100 mL). The combined organic layers were washed with brine (2X 100mL) and Na2SO4Drying and evaporating under reduced pressure. The crude residue was purified by column chromatography (100-200 silica) using 20% ethyl acetate/petroleum ether to give ethyl 3- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) propionate 4 as a pale yellow oily liquid (3.4g, 11.72mmol, 40% yield). TLC system: 40% ethyl acetate in petroleum ether, Rf：0.50
3- (2- (2-Hydroxyethoxy) ethoxy) propionic acid ethyl ester (5)
To a stirred solution of ethyl 3- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) propionate 4(4.3g, 14.827mmol) in methanol (40mL) at 0 ℃ was added pyripyropene p-toluenesulfonate (1.86g, 7.413mmol) and stirred at room temperature for 16H. The solvent was distilled off under reduced pressure to give a residue, which was diluted with water (50mL) and extracted with ethyl acetate (3X 50 mL). The combined organics were washed with brine (50mL), dried over sodium sulfate and concentrated. The crude residue was purified by column chromatography (100-200 silica) using 70% ethyl acetate in petroleum ether to give ethyl 3- (2- (2-hydroxyethoxy) ethoxy) propionate 5 as a pale yellow gummy liquid (2.6g, 12.62mmol, 86% yield). TLC system: 70% ethyl acetate in petroleum ether, R f：0.20
3- (2- (2- (2-tert-butoxy-2-oxoethoxy) ethoxy) propionic acid ethyl ester
A stirred solution of ethyl 3- (2- (2-hydroxyethoxy) ethoxy) propionate 5(2.0g, 9.708mmol) in N, N-dimethylformamide (20mL) was treated with NaH (0.35g, 14.65mmol) at 0 deg.C and stirred at room temperature for 30 min. Tert-butyl 2-bromoacetate (2.27g, 11.65mmol) in N, N-dimethylformamide (10mL) was added to the above reaction mixture at 0 ℃ and stirred at room temperature under a nitrogen atmosphere for 3 hours. The reaction mixture was quenched with ice water (50mL) and extracted with ethyl acetate (3X 50 mL). The combined organic layers were washed with brine (2X 50mL) and Na2SO4Drying and evaporating under reduced pressure. The crude residue was purified by column chromatography (100-200 silica) using 20% ethyl acetate in petroleum ether to give ethyl 3- (2- (2- (2-tert-butoxy-2-oxoethoxy) ethoxy) propanoate 6(600mg, 1.875mmol, 19% yield) as a pale yellow oily liquid. TLC system: 70% ethyl acetate in petroleum ether, Rf：0.60
12-oxo-3, 6,9, 13-tetraoxapentadecane-1-oic acid (side chain-7)
To a stirred solution of ethyl 3- (2- (2- (2-tert-butoxy-2-oxoethoxy) ethoxy) propionate 6(600mg, 1.875mmol) in dioxane (5mL) was added a 4N HCl solution in dioxane (1mL) followed by stirring at room temperature for 16 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, the crude material was diluted with water (50mL) and extracted into ethyl acetate (2 × 30 mL). The organic layer was washed with brine solution (30mL) and Na 2SO4Dried and concentrated. The crude compound was purified by washing with diethyl ether (30mL) to obtain 12-oxo-3, 6,9, 13-tetraoxapentadecan-1-oic acid (side chain-7) (4.0g, 10.69mmol, 85%) as a pale yellow oily liquid. TLC system: 10% methanol in dichloromethane, Rf：0.20
2, 2-dimethyl-4-oxo-3, 6,9, 12-tetraoxapentadecane-15-oic acid (side chain-8)
To ethyl 3- (2- (2- (2- (tert-butoxy) -2-oxoethoxy) ethoxy) propionate (200mg, 0.625mmol, the synthesis of compound 1 is reported in side chain-7) in THF at 0 ℃: h2To a stirred solution of O mixture (4:1) (10mL) was added lithium hydroxide (26mg, 0.625mmol), followed by stirring at room temperature for 4 hours. The reaction mixture was concentrated to remove organic volatiles, water (50mL) was added and extracted with ethyl acetate (2 × 30 mL). The combined organic layers were washed with brine solution (20mL), dried over sodium sulfate and concentrated under reduced pressure. The crude residue was purified by column chromatography eluting with 20% ethyl acetate in petroleum ether to give 2, 2-dimethyl-4-oxo-3, 6,9, 12-tetraoxapentadecane-15-oic acid side chain-8 (70mg, 0.239mmol, 38%) as a colorless oily liquid. TLC system: 40% ethyl acetate in petroleum ether, R f：0.40
Side chain-9
2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethan-1-ol (2)
To a stirred solution of 2,2' - ((oxybis (ethane-2, 1-diyl)) bis (oxy)) bis (ethan-1-ol) 1(10g, 51.546mmol) in dichloromethane (100mL) was added dihydropyran (2.8mL, 30.6mmol) and p-toluenesulfonic acid (979mg, 5.154mmol) at 0 ℃ and stirred at room temperature for 4 hours. The reaction mixture was diluted with water (100mL), extracted with dichloromethane (3 × 200mL), the combined organic layers were washed with brine (2 × 75mL), dried over sodium sulfate and concentrated. The residue was purified by flash column chromatography (100-200 silica) using 2% methanol/dichloromethane to give 2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethan-1-ol 2(2.2g, 7.913mmol, 15.4% yield) as a yellow thick liquid. TLC system: 10% methanol in dichloromethane, Rf：0.20
4-Methylbenzenesulfonic acid 2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl ester (3)
To a stirred solution of 2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethan-1-ol 2(1g, 4.27mmol) in DCM (20mL) at 0 deg.C was added Et 3N (0.8ml, 5.55mmol) and p-toluenesulfonyl chloride (1.05g, 5.55mmol), and the reaction mixture was stirred at room temperature for 3 hours. After the starting material was consumed, the mixture was diluted with ethyl acetate (50mL) and ice-cold water (2X 20mL), NaHCO3The solution (2X 20mL), brine solution (20mL) was washed, dried over sodium sulfate and concentrated. The crude compound was purified by combined flash chromatography and eluted with 50% ethyl acetate in hexanes to give 4-methylbenzenesulfonic acid 2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl ester 3(1g, 2.31mmol, 54% yield) as a light brown liquid. TLC system: 100% ethyl acetate, Rf：0.50；LCMS：m/z＝455.39(M+Na)+
2- (2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl) isoindoline-1, 3-dione (4)
To a stirred solution of 4-methylbenzenesulfonic acid 2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl ester 3(500Mg, 1.15mmol) in DMF (10mL) was added potassium phthalimide (282Mg, 1.52mmol) at room temperature, followed by stirring at 110 ℃ for 16 hours. After completion of the reaction as indicated by TLC, ice water (2 × 200mL) was added to the reaction mixture and extracted with ethyl acetate (2 × 100 mL). The organic layer was washed with brine solution (100mL) over Na 2SO4Drying and concentrating to obtain 2- (2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl) isoindoline as a light brown liquid-1, 3-dione 4(400mg, 9.828mmol, 84%). TLC system: 50% ethyl acetate in hexane, Rf：0.20；LCMS：m/z＝429.97(M+Na)+
2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethan-1-amine (side chain-9)
To a stirred solution of 2- (2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl) isoindoline-1, 3-dione 4(400g, 0.98mmol) in ethanol (3mL) was added hydrazine hydrate (3mL) and stirred at 110 ℃ for 12 hours. After completion of the reaction as indicated by TLC, toluene (10ml) was added to the reaction mixture, two layers were formed and separated. The toluene layer was washed with brine solution (10mL) and Na2SO4Dried and concentrated to give 2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethan-1-amine side chain-9 (150mg, 0.541mmol, 55%) as a brown liquid. TLC system: 10% methanol in dichloromethane, Rf：0.20
Side chain-10
N-methyl-2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethan-1-amine (side chain-10)
To a stirred solution of 4-methylbenzenesulfonic acid 2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethyl ester 1(500mg, 1.15mmol, the synthesis of compound-1 is reported in side chain-9) in a sealed tube at room temperature was added 33% MeNH in ethanol 2(5mL), followed by stirring at 60 ℃ for 12 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, sodium bicarbonate solution (20mL) was added to the residue and extracted with 10% methanol in dichloromethane (3 × 20 mL). The combined organic layers were washed with brine solution (100mL) and dried over anhydrous Na2SO4Drying and concentration gave N-methyl-2- (2- (2- (2- ((tetrahydro-2H-pyran-2-yl) oxy) ethoxy) ethan-1-amine side chain-10 (150mg, 0.515mmol, 44%) as a brown liquid. TLC system: 10% methanol in dichloromethane, Rf：0.20
Side chain-11
2, 2-dimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azaheptadecane-17-oic acid ethyl ester (3)
A stirred solution of tert-butyl (2- (2- (2-hydroxyethoxy) ethoxy) ethyl) carbamate 1(1g, 4.01mmol) in N, N-dimethylformamide (15mL) was treated with NaH (240mg, 10.04mmol) at 0 deg.C to room temperature for 30 minutes. Ethyl 3-bromopropionate 2(1.1g, 6.02mmol) in N, N-dimethylformamide was added to the above reaction mixture at 0 ℃ and stirred at room temperature under an argon atmosphere for 16 hours. The reaction mixture was quenched with ice water and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with brine (50mL) and Na 2SO4Drying and evaporating under reduced pressure. The crude residue was purified by column chromatography eluting with 70% ethyl acetate in petroleum ether to give ethyl 2, 2-dimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azaheptadecane-17-carboxylate 3(310mg, 0.925mmol, 24% yield) as a colorless liquid. TLC system: 70% ethyl acetate in petroleum ether, Rf: 0.50; direct quality: m/z 372.15(M + Na)+
3- (2- (2- (2-aminoethoxy) ethoxy) propionic acid ethyl ester (side chain-11)
To a stirred solution of ethyl 2, 2-dimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azaheptadecane-17-carboxylate 3(310mg, 0.888mmol) in dioxane (1mL) was added HCl in dioxane (4M, 3mL) at room temperature and stirred under an argon atmosphere for 16 hours. The organic solvent was distilled off and co-distilled with dichloromethane to give ethyl 3- (2- (2- (2-aminoethoxy) ethoxy) propanoate side chain 11 as a colorless liquid (260mg, 1.044mmol, crude). TLC system: 20% methanol in dichloromethane, Rf: 0.10; direct quality: m/z 250.12(M + H)+
Side chain-12
3- (2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) propionic acid ethyl ester (3)
A stirred solution of 2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) ethanol 1 (the synthesis of compound 1 is reported in side chain-5) (5g, 21.367mmol) in N, N-dimethylformamide (30mL) was treated with NaH (1.3g, 53.417mmol) at 0 ℃ and stirred at room temperature for 30 minutes. Ethyl 3-bromopropionate 2(5.8g, 32.05mmol) in N, N-dimethylformamide (20mL) was added to the above reaction mixture at 0 ℃ and stirred at room temperature under a nitrogen atmosphere for 16 hours. The reaction mixture was quenched with ice water and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine (2X 100mL) and Na 2SO4Drying and evaporating under reduced pressure. The crude residue was purified by combined flash chromatography using 30% ethyl acetate in hexanes to give ethyl 3- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) propanoate 3(1g, 2.994mmol, 14% yield) as a light yellow oily liquid. TLC system: 70% ethyl acetate in hexane; rf：0.60
3- (2- (2- (2-hydroxyethoxy) ethoxy) propionic acid ethyl ester (4)
To a stirred solution of ethyl 3- (2- (2- (2- (tetrahydro-2H-pyran-2-yloxy) ethoxy) propionate 3(1g, 2.994mmol) in methanol (10mL) at 0 ℃ was added pyripyropene p-toluenesulfonate (376mg, 1.497mmol) and stirred at room temperature for 16H. The volatiles were removed under reduced pressure, water (50mL) was added and extracted with ethyl acetate (2X 30 mL). The combined organic layers were washed with brine solution (30mL), dried over sodium sulfate and concentrated to give ethyl 3- (2- (2- (2-hydroxyethoxy) ethoxy) propanoate 4 as a pale yellow oily liquid (580mg, 2.32mmol, 77% yield). TLC system: 5% methanol in dichloromethane, Rf：0.20
3- (2- (2- (2- (tosyloxy) ethoxy) propionic acid ethyl ester (5)
To a stirred solution of ethyl 3- (2- (2- (2-hydroxyethoxy) ethoxy) propionate 4(580mg, 2.32mmol) in THF (5mL) at 0 ℃ was added NaOH (186mg, 4.64mmol) and stirred at room temperature for 15 min. Then cooled to 0 ℃, p-toluenesulfonyl chloride (530mg, 2.784mmol) was added to the reaction mixture and stirred at room temperature for 16 hours. After the starting material was consumed according to TLC, the reaction mixture was diluted with ethyl acetate (50mL) and washed with ice-cold water (2 × 40mL), brine solution (20mL), dried over sodium sulfate and concentrated to give ethyl 3- (2- (2- (2- (tosyloxy) ethyl) ethoxy) propionate 5(750mg, 1.856mmol, 17% yield) as a light yellow oily liquid. TLC system: 70% ethyl acetate in hexane; r f：0.70
5,8, 11-Trioxa-2-azatetradecane-14-oic acid ethyl ester (side chain-12)
Ethyl 3- (2- (2- (2- (tosyloxy) ethoxy) propionate 5(750mg, 1.856mmol) was placed in a sealed tube and dissolved in ethanol (5 mL). Addition of MeNH containing 33% in ethanol at room temperature2(0.9mL, 9.282mmol) and then stirred at 60 ℃ for 16 h. After completion of the reaction as indicated by TLC, volatiles were removed under reduced pressure, water (50mL) was added and acidified with 1N HCL solution to pH 2. The aqueous layer was extracted with ethyl acetate (50mL), brine solution (30mL), and Na2SO4Drying and concentration gave side chain-12 as a reddish brown gummy liquid (200mg, 0.7604mmol, 41%). TLC system: 10% methanol in dichloromethane, Rf：0.10
Side chain-13
4-Methylbenzenesulfonic acid 2- (2- (2- (2-hydroxyethoxy) ethoxy) ethyl ester (2)
To a stirred solution of 2,2' - ((oxybis (ethane-2, 1-diyl)) bis (oxy)) bis (ethan-1-ol) 1(2g, 10.30mmol) in dichloromethane (30mL) was added silver oxide (3.5g, 15.46mmol), p-toluenesulfonyl chloride (2.3g, 12.37mmol), potassium iodide (342mg, 2.06mmol) at 0 ℃. The mixture was stirred at 0 ℃ for 30 minutes. After the starting material was consumed, the mixture was filtered through a pad of celite and the pad was washed several times with DCM. The filtrate was washed with water, brine solution (50mL), dried over sodium sulfate and concentrated. The crude compound was purified by column chromatography and eluted with 3% methanol in dichloromethane to give 2- (2- (2- (2-hydroxyethoxy) ethoxy) ethyl 4-methylbenzenesulfonate 2(2.6g, 7.471mmol, 72% yield) as a colorless oily liquid. TLC system: 10% to two Methanol in methyl chloride, Rf：0.50；LCMS：m/z＝371.19(M+Na)+
5,8, 11-trioxa-2-azatridecan-13-ol (3)
To a stirred solution of 4-methylbenzenesulfonic acid 2- (2- (2- (2-hydroxyethoxy) ethoxy) ethyl ester 2(600mg, 1.724mmol) in ethanol (5mL) at room temperature in a sealed tube was added 33% MeNH in ethanol2(0.8mL) (267mg, 8.62mmol), followed by stirring at 60 ℃ for 16 h. After completion of the reaction as indicated by TLC, the organic solvent was evaporated and the crude material co-distilled with dichloromethane (10mL) to give 5,8, 11-trioxa-2-azatridecan-13-ol 3 as a colorless gummy liquid (650mg, 3.14mmol, crude material). TLC system: 10% methanol in dichloromethane, Rf0.20; direct quality: m/z 208.15(M + H)+
(2- (2- (2- (2-hydroxyethoxy) ethoxy) ethyl) (methyl) carbamic acid tert-butyl ester (4)
To a stirred solution of 5,8, 11-trioxa-2-azatridecan-13-ol 3(2.9g, 14.09mmol) in DCM (30mL) at 0 deg.C were added triethylamine (3.8mL, 28.01mmol) and di-tert-butyl dicarbonate (3.2mL, 14.09mmol), followed by stirring at room temperature for 2 hours. After the starting material was consumed, the mixture was diluted with ethyl acetate (150mL) and washed with ice-cold water (2 × 20mL), brine solution (20mL), dried over sodium sulfate and concentrated under reduced pressure. The crude compound was purified by 100-phase 200 silica gel column chromatography eluting with 5% methanol in dichloromethane to give tert-butyl (2- (2- (2- (2-hydroxyethoxy) ethoxy) ethyl) (methyl) carbamate 4(1.8g, 5.863mmol, 42% yield) as a light brown liquid. TLC system: 100% ethyl acetate, R f：0.50；LCMS：m/z＝330.04(M+Na)+
4-Methylbenzenesulfonic acid 2,2, 5-trimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azahexadecan-16-yl ester (5)
To a stirred solution of tert-butyl (2- (2- (2- (2-hydroxyethoxy) ethoxy) ethyl) (methyl) carbamate 4(1.85g, 6.026mmol) in THF (20mL) was added sodium hydroxide (485mg, 12.052mmol) and p-toluenesulfonyl chloride (1.63g, 7.231mmol) at 0 deg.C and stirred at room temperature for 16 h. After completion of the reaction as indicated by TLC, the mixture was diluted with ethyl acetate (150mL) and washed with ice-cold water (2 × 20mL), brine solution (20mL), dried over sodium sulfate and concentrated under reduced pressure. The crude compound was purified by 100-200 silica gel column chromatography eluting with 40% ethyl acetate in petroleum ether to give 2,2, 5-trimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azahexadecan-16-yl 4-methylbenzenesulfonate 5(2.5g, 5.423mmol, 90% yield) as a colorless gum. TLC system: 70% ethyl acetate in petroleum ether, Rf：0.50；LCMS：m/z＝484.4(M+Na)+
(2- (2- (2- (2- (1, 3-dioxoisoindolin-2-yl) ethoxy) ethyl) (methyl) carbamic acid tert-butyl ester (6)
To a stirred solution of 4-methylbenzenesulfonic acid 2,2, 5-trimethyl-4-oxo-3, 8,11, 14-tetraoxa-5-azahexadecan-16-ester 5(2.5g, 5.423mmol) in DMF (10mL) was added potassium phthalimide (1.3g, 7.049mmol) at room temperature, followed by stirring at 110 ℃ for 16 hours. The reaction mixture was allowed to cool to room temperature, then diethyl ether (100mL) was added and stirred for 15 minutes, filtered. The filtrate was washed with 1M NaOH solution (50mL), water (100mL), brine solution (100mL), and Na 2SO4Dried and concentrated under reduced pressure to give (2- (2- (2- (1, 3-dioxoiso-2) as a pale brown gummy liquidIndolin-2-yl) ethoxy) ethyl) (methyl) carbamic acid tert-butyl ester 6(1.65g, 3.784mmol, crude). TLC system: 70% ethyl acetate in petroleum ether, Rf：0.50；LCMS：m/z＝459.45(M+Na)+
2- (5,8, 11-trioxa-2-azatridecan-13-yl) isoindoline-1, 3-dione (side chain-13)
To a stirred solution of tert-butyl (2- (2- (2- (2- (1, 3-dioxoisoindolin-2-yl) ethoxy) ethyl) (methyl) carbamate 6(1.55g, 3.555mmol) in dioxane (5mL) was added 4N HCl in dioxane (5mL) followed by stirring at room temperature for 30 minutes. The reaction mixture was concentrated, water (50mL) was added to the crude, basified with sodium bicarbonate solution (100mL), and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with brine solution (50mL) and Na2SO4Drying and concentration under reduced pressure gave 2- (5,8, 11-trioxa-2-azatridecan-13-yl) isoindoline-1, 3-dione side chain-13 (700mg, 2.083mmol, crude material) as a pale yellow oily liquid. TLC system: 70% ethyl acetate in petroleum ether, Rf：0.10；LCMS：m/z＝337.41(M+H)+
Side chain-14
5- (2-ethoxyethoxy) pentanoic acid ethyl ester (3)
To a stirred solution of 2-ethoxyethanol 1(500mg, 5.555mmol) in tetrahydrofuran (10mL) at 0 deg.C was added NaH (267mg, 11.11mmol) and stirred at room temperature for 30 min. In tetrahydrofuran (5mL) at 0 deg.CEthyl 5-bromovalerate 2(1.74g, 8.333mmol) was added to the above reaction mixture and stirred at room temperature under a nitrogen atmosphere for 16 hours. The reaction mixture was quenched with ice-cold water (50mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with brine (50mL) and Na2SO4Dried and evaporated under reduced pressure. The crude residue was purified by 100-200 silica gel chromatography using 6% ethyl acetate in hexane to give ethyl 5- (2-ethoxyethoxy) pentanoate 3(220mg, 1.009mmol, 18% yield) as a colorless oily liquid. TLC system: 50% ethyl acetate in hexane; rf:0.45
5- (2-ethoxyethoxy) pentanoic acid (side chain-14)
To ethyl 5- (2-ethoxyethoxy) pentanoate 3(220mg, 1.009mmol) in THF: h2To a stirred solution of O in mixture (3:1) (4mL) was added lithium hydroxide monohydrate (127mg, 3.027mmol) and stirred for 16 h. The reaction mixture was concentrated to remove organic volatiles, water (50mL) was added to the crude and extracted with ethyl acetate (2 × 20 mL). The aqueous solution was acidified with a saturated citric acid solution and extracted with 10% methanol in dichloromethane (2 × 30 mL). The combined organic layers were washed with brine solution (15mL), dried over sodium sulfate and concentrated to give 5- (2-ethoxyethoxy) pentanoic acid side chain-14 (110mg, 0.5789mmol, 57%) as a colorless oily liquid. TLC system: 70% ethyl acetate in hexane; r f：0.10
Side chain-15
5- (2-Ethoxyethoxy) pentan-1-ol (3)
A stirred solution of 1, 5-pentanediol 1(11g, 105.76mmol) in N, N-dimethylformamide (30mL) was treated with 60% NaH (4.5g, 116.34mmol) at 0 deg.C and stirred at room temperature for 30 min. 1-bromo-2-ethoxyethane 2(12mL, 105.76mmol) in N, N-dimethylformamide (20mL) was added to the above reaction mixture at 0 deg.C and stirred at room temperature under an argon atmosphere for 16 hours. The reaction mixture was quenched with ice water (200mL) and extracted with ethyl acetate (3X 250 mL). The combined organic layers were washed with brine (2X 100mL) and Na2SO4Drying and evaporating under reduced pressure. The crude residue was purified by column chromatography (100-200 silica gel) using 30% ethyl acetate in hexane to give 5- (2-ethoxyethoxy) pentan-1-ol 3 as an oily liquid (5.1g, 28.97mmol, 27% yield). TLC system: 70% ethyl acetate in hexane, Rf：0.50；LCMS：m/z＝199.12(M+Na)+
4-Methylbenzenesulfonic acid 5- (2-ethoxyethoxy) pentyl ester (4)
To a stirred solution of 5- (2-ethoxyethoxy) pentan-1-ol 3(4.6g, 26.136mmol) in DCM (45mL) was added triethylamine (11g, 78.40mmol) at 0 ℃. The mixture was stirred at 0 ℃ for 10 minutes. Then cooled to 0 ℃, p-toluenesulfonyl chloride (5.95g, 31.36mmol) was added and DMAP (31mg, 0.261mmol) was added to the reaction mixture and stirred at room temperature for 16 hours. After the starting material was consumed, the mixture was diluted with dichloromethane (100mL) and washed with ice-cold water (2 × 50mL), brine solution (50mL), dried over sodium sulfate and concentrated under reduced pressure. The crude compound was purified by combined flash chromatography eluting with 35% ethyl acetate in petroleum ether to give 4-methylbenzenesulfonic acid 5- (2-ethoxyethoxy) pentyl ester 4(3.9g, 11.81mmol, 45% yield) as a yellow oily liquid. TLC system: 50% ethyl acetate in hexane, R f：0.60；LCMS：m/z＝353.20(M+Na)+
1-azido-5- (2-ethoxyethoxy) pentane (5)
To a stirred solution of 4-methylbenzenesulfonic acid 5- (2-ethoxyethoxy) pentyl ester 4(1.4g, 4.242mmol) in DMF (15mL) at room temperature was added NaN3(441mg, 6.787mmol), followed by stirring at 60 ℃ for 16 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, water (200mL) was added to the crude material and extracted with ether (2 × 50 mL). The combined organic layers were washed with brine solution (50mL) and Na2SO4Dried and concentrated. The crude compound was purified by column chromatography (silica gel 100-200) eluting with 30% ethyl acetate in petroleum ether to give 1-azido-5- (2-ethoxyethoxy) pentane 5(750mg, 3.731mmol, 88%) as a liquid. TLC system: 30% ethyl acetate in hexane, Rf:0.40
5- (2-ethoxyethoxy) pentan-1-amine (side chain-15)
To 1-azido-5- (2-ethoxyethoxy) pentane 5(840mg, 4.179mmol) in THF at 0 deg.C: h2To a stirred solution of O (4:1) (10mL) was added 1M P (Me) in THF3(8.3mL, 8.358mmol), followed by stirring at room temperature for 16 h. After completion of the reaction as indicated by TLC, the organic solvent was evaporated to give 5- (2-ethoxyethoxy) pentan-1-amine side chain-15 (1.1g, crude material) as a liquid. TLC system: 10% methanol in dichloromethane, R f：0.10；LCMS：m/z＝176.18(M+H)+
5- (2-ethoxyethoxy) -N-methylpent-1-amine (side chain-16)
Stirring of 4-methylbenzenesulfonic acid 5- (2-ethoxyethoxy) pentyl ester 1 (synthesis of compound 1 is reported in side chain-15) (3) at 70 ℃g, 9.090mmol) in a solution of 1M methylamine in ethanol (70mL) for 6 h. After completion of the reaction as indicated by TLC, the reaction mixture was evaporated and diluted with ethyl acetate (150 mL). The organic layer was basified with triethylamine, water was added and extracted with ethyl acetate (2X 100 ml). The combined organic layers were washed with brine solution (50mL) and Na2SO4Dried and concentrated to give 5- (2-ethoxyethoxy) -N-methylpentan-1-amine side chain-16 as a brown oily liquid (1.6g, 8.465mmol, crude material). TLC system: 10% methylene chloride/triethylamine, Rf: 0.20; direct quality: 190(M + H) M/z+
Side chain-17
N-benzyl-5- (2-ethoxyethoxy) pentanamide (2)
To a stirred solution of 5- (2-ethoxyethoxy) pentanoic acid 1 (synthesis of compound 1 is reported in side chain-14) (500mg, 2.64mmol) in DMF (5mL) at room temperature was added diisopropylethylamine (1.4mL, 7.9mmol), HATU (2g, 5.28mmol) and stirred for 10 min, followed by addition of benzylamine (420mg, 3.96mmol) and stirring at room temperature for 16 h. After completion of the reaction as indicated by TLC, ice-cold water (50mL) was added to the reaction mixture and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with brine solution (50mL) and dried (Na) 2SO4) And concentrated. The crude compound was purified by combined flash chromatography and eluted with 25% ethyl acetate in petroleum ether to give N-benzyl-5- (2-ethoxyethoxy) pentanamide 2 as a pale yellow liquid (400mg, 1.43mmol, 54% yield). TLC system: 50% ethyl acetate in hexane, Rf：0.40；LCMS：m/z＝280.39(M+H)+
N-benzyl-5- (2-ethoxyethoxy) pentan-1-amine (side chain 17)
To a stirred solution of N-benzyl-5- (2-ethoxyethoxy) pentanamide 2(400mg, 1.43mmol) in THF (5mL) at 0 deg.C was added a solution of 1M borane in THF (5.7mL, 5.73mmol) and the mixture was stirred at room temperature for 4 h. After completion of the reaction as indicated by TLC, excess borane was carefully quenched with 1mL methanol, ice water (50mL) and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with brine (50mL) and Na2SO4Drying and evaporation under reduced pressure gave N-benzyl-5- (2-ethoxyethoxy) pentan-1-amine side chain 17 as a pale yellow liquid (300mg, 1.13mmol, 79% yield). TLC system: 50% ethyl acetate in hexane, Rf：0.50；LCMS：m/z＝288.14(M+Na)+
Side chain-18
5-ethoxypentan-1-ol (2)
To a stirred solution of pentane-1, 5-diol 1(500mg, 4.807mmol)) in tetrahydrofuran (10mL) at 0 deg.C was added NaH (115mg, 4.807mmol) and stirred at room temperature for 1 hour. Iodoethane (740mg, 4.807mmol) in tetrahydrofuran (5mL) was added to the above reaction mixture at 0 ℃ and stirred at 60 ℃ under a nitrogen atmosphere for 48 hours. The reaction mixture was quenched with ice water and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with brine (20mL) and Na 2SO4Dried and evaporated under reduced pressure. The crude residue was purified by 100-200 silica gel chromatography using 35% ethyl acetate in hexane to give 5-ethoxypentan-1-ol 2 as a red-brown oily liquid (310mg, 2.348mmol, 49% yield). TLC system: 70% of ethyl in hexaneEthyl ester of acid; rf：0.50
2- (5-Ethoxypentyloxy) acetic acid tert-butyl ester (4)
To a stirred solution of 5-ethoxypentan-1-ol 2(500mg, 3.7878mmol) in N, N-dimethylformamide (5mL) were added potassium carbonate (1.6g, 11.363mmol) and tert-butyl 2-bromoacetate 3(1.85g, 9.4697mmol), followed by stirring at 80 ℃ under a nitrogen atmosphere for 16 hours. The reaction mixture was quenched with ice water and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with brine solution (20mL) and Na2SO4Dried and evaporated under reduced pressure. The crude residue was purified by 100-200 silica gel chromatography using 8% ethyl acetate in hexane to give tert-butyl 2- (5-ethoxypentoxy) acetate 4(600mg, 2.439mmol, 64% yield) as a colorless oily liquid. TLC system: 30% ethyl acetate in hexane, Rf：0.50
2- (5-Ethoxypentyloxy) acetic acid (side chain-18)
To a stirred solution of tert-butyl 2- (5-ethoxypentoxy) acetate 4(2g, 8.13mmol)) in dioxane (10mL) was added 4N HCl in dioxane (15mL), followed by stirring at room temperature for 4 hours. The reaction mixture was concentrated, water (100mL) was added to the crude material and extracted with ethyl acetate (2 × 50 mL). The aqueous layer was acidified with 1N HCl solution to pH 2 and then extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with brine solution (100mL) and Na 2SO4Drying and concentration gave 2- (5-ethoxypentoxy) acetic acid side chain-18 (1.3g, 6.842mmol, 83%) as a colorless, gummy liquid. TLC system: 100% ethyl acetate, Rf：0.10
Side chain-19
5-ethoxypentan-1-ol (2)
To a stirred suspension of NaH (3.8g, 96.15mmol) was added a solution of pentane-1, 5-diol 1(10g, 96.15mmol) in THF (100mL) at 0 deg.C and stirred at room temperature for 1 hour. Iodoethane (3.7mL, 96.15mmol) in THF (50mL) was added to the above reaction mixture at 0 ℃ and stirred at room temperature under an argon atmosphere for 16 h. The reaction mixture was quenched with ice-cold water (200mL) and extracted with ethyl acetate (3 × 200 mL). The combined organic layers were washed with brine (100mL) and Na2SO4Dried and evaporated under reduced pressure. The crude residue was purified by column chromatography eluting with 40% ethyl acetate in petroleum ether to give 5-ethoxypentan-1-ol 2 as an oily liquid (6.5g, 49.24mmol, 51% yield). TLC system: 70% ethyl acetate in petroleum ether, Rf: 0.50; direct quality: m/z 133.36(M + H)+
((2-Bromoethoxy) methanetrityl) triphenyl (4)
To a stirred solution of 2-bromoethan-1-ol 3(7g, 56mmol) in DCM (175mL) at 0 deg.C was added triethylamine (15.5mL, 112mmol) and trityl chloride and stirred at room temperature for 3 h. After completion of the reaction as indicated by TLC, the reaction mixture was diluted with water (300mL) and extracted with dichloromethane (2 x 200 mL). The combined organic layers were washed with brine solution (100mL) and Na 2SO4Dried and concentrated under reduced pressure. The crude compound was purified by 100-200 silica gel column chromatography eluting with 5% ethyl acetate in petroleum ether to give ((2-bromoethoxy) methane trityl) triphenyl 4(4.5g, 12.295mmol, 22% yield) as an off-white solid. TLC system: 10 percent ofEthyl acetate in Petroleum Ether, Rf：0.60；LCMS：m/z＝367.24(M+H)+
((2- ((5-ethoxypentyl) oxy) ethoxy) methane trityl) triphenyl (5)
To a stirred suspension of NaH (568mg, 23.67mmol)) was added a solution of 5-ethoxypentan-1-ol 2(1.25g, 9.469mmol) in DMF (20mL) at 0 ℃ and stirred at room temperature for 30 min. ((2-bromoethoxy) methanetrityl) triphenyl 4(4.5g, 12.31mmol) in DMF (5mL) is then added to the above reaction mixture at 0 ℃ and stirred at room temperature under an argon atmosphere for 16 hours. The reaction mixture was quenched with ice-cold water (200mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine (100mL) and Na2SO4Dried and evaporated under reduced pressure. The crude residue was purified by 100-pack 200 silica gel column chromatography eluting with 5% ethyl acetate in petroleum ether to give ((2- ((5-ethoxypentyl) oxy) ethoxy) methane trityl triphenyl 5(2.5g, 5.980mmol, 63% yield) as a colorless oily liquid a TLC system 5% ethyl acetate in petroleum ether, R f：0.40；LCMS：m/z＝441.46(M+Na)+
2- ((5-ethoxypentyl) oxy) ethan-1-ol (6)
To a stirred solution of ((2- ((5-ethoxypentyl) oxy) ethoxy) methane trityl) triphenyl 5(12g, 28.70mmol) in dioxane (60mL) was added HCl in dioxane (4N, 24mL) at 0 ℃, followed by stirring at room temperature for 1 hour. After completion of the reaction as indicated by TLC, the reaction mixture was concentrated, water (50mL) was added to the residue, basified with sodium bicarbonate solution (100mL) and extracted with ethyl acetate (2 × 200 mL). The combined organic layers were washed with brine solution (100mL) and Na2SO4Dried and concentrated under reduced pressure. The crude compound was purified by 100-pack 200 silica gel column chromatography eluting with 70% ethyl acetate in petroleum ether to give 2- ((5-ethoxypentyl) oxy) ethan-1-ol 6(4.4g, 25.0mmol, 88% yield) as a colorless oily liquid. TLC system: 50% ethyl acetate in petroleum ether, Rf: 0.30; direct quality: m/z 177.22(M + H)+
4-Methylbenzenesulfonic acid 2- ((5-ethoxypentyl) oxy) ethyl ester (7)
To a stirred solution of 2- ((5-ethoxypentyl) oxy) ethan-1-ol 6(3g, 17.04mmol) in THF (5mL) at 0 deg.C was added NaOH (1.3g, 34.09mmol) and p-toluenesulfonyl chloride (3.8g, 20.45mmol) to the reaction mixture and stirred at room temperature for 16 h. After the starting material was consumed, the reaction mixture was quenched with water (200mL) and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with brine solution (50mL), dried over sodium sulfate and concentrated under reduced pressure. The crude compound was purified by 100-200 silica gel column chromatography eluting with 20% ethyl acetate in petroleum ether to give 2- ((5-ethoxypentyl) oxy) ethyl 4-methylbenzenesulfonate 7(5.2g, 15.75mmol, 92% yield) as a liquid. TLC system: 20% ethyl acetate in petroleum ether, R f：0.50；LCMS：m/z＝331.04(M+H)+
1- (2-azidoethoxy) -5-ethoxypentane (8)
To a stirred solution of 4-methylbenzenesulfonic acid 2- ((5-ethoxypentyl) oxy) ethyl ester 7(3.5g, 10.60mmol) in DMF (30mL) at room temperature was added NaN3(1.1g, 16.96mmol), followed by stirring at 60 ℃ for 16 hours. After completion of the reaction as indicated by TLC, the reaction mixture was quenched with water (200mL) and extracted with ether (2 × 100 mL). The combined organic layers were washed with brine solution (100mL) and Na2SO4Dried and concentrated under reduced pressure. The crude compound was purified by column chromatography eluting with 15% ethyl acetate in petroleum ether to give 1- (2-azidoethoxy) -5-ethoxypentane 8(1.95g, 6.414mmol, 92%) as a colorless oily liquid. TLC system: 20% ethyl acetate in petroleum ether, Rf：0.50
2- ((5-ethoxypentyl) oxy) ethan-1-amine (side chain-19)
To 1-azido-5- (2-ethoxyethoxy) pentane 8(900mg, 4.477mmol) in THF at 0 deg.C: h2To a stirred solution of O (4:1) (12.5mL) was added 1M P (Me) in THF3(8.9mL, 8.955mmol), followed by stirring at room temperature for 16 h. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, water (100mL) was added to the residue and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with brine solution (100mL), dried over sodium sulfate and concentrated under reduced pressure to give 5- (2-ethoxyethoxy) pentan-1-amine side chain-19 (240mg, 1.371mmol, crude material) as a pale yellow oily liquid. TLC system: 5% methanol in dichloromethane, R f: 0.10; direct quality: m/z 176.31(M + H)+
Side chain-20
2- ((5-ethoxypentyl) oxy) -N-methylethyl-1-amine (side chain-20)
To a stirred solution of 4-methylbenzenesulfonic acid 2- ((5-ethoxypentyl) oxy) ethyl ester 1 (the synthesis of compound 1 is reported in side chain-19) (1g, 3.030mmol) in ethanol (3mL) in a sealed tube at room temperature was added MeNH in ethanol2(1M, 5mL), followed by stirring in a sealed tube at 60 ℃ for 16 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated. To the crude material was added water (10mL), acidified with HCl solution (6N, 50mL) and dichloromethaneAlkane (2X 100 mL). The aqueous layer was basified with NaOH solution (100mL) and extracted with ethyl acetate (2X 100 mL). The organic layer was washed with brine solution (50mL) over Na2SO4Drying and concentration gave 2- ((5-ethoxypentyl) oxy) -N-methylethyl-1-amine side chain-20 (240mg, 1.269mmol, 41%) as a colorless oily liquid. TLC system: 70% ethyl acetate in petroleum ether, Rf: 0.10; direct quality: m/z 190.18(M + H)+
Side chain-21
N-benzyl-2- ((5-ethoxypentyl) oxy) ethan-1-amine (side chain-21)
To a stirred solution of 4-methylbenzenesulfonic acid 2- ((5-ethoxypentyl) oxy) ethyl ester 1 (the synthesis of compound 1 is reported in side chain-19) (65mg, 0.196mmol) in ethanol (4mL) in a sealed tube was added benzylamine (63mg, 0.5908mmol) at room temperature, followed by stirring at 70 ℃ for 16 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, water (50mL) was added to the crude material and extracted with ethyl acetate (2 × 20 mL). The combined organic layers were washed with brine solution (30mL) and Na 2SO4Drying and concentration gave the crude compound, N-benzyl-2- ((5-ethoxypentyl) oxy) ethan-1-amine side chain-21 (95mg, 0.358mmol, crude material) as a colorless, gummy liquid. TLC system: 10% methanol in dichloromethane, Rf：0.40；LCMS：m/z＝266.29(M+H)+
Side chain-22
2- (pentyloxy) ethan-1-ol (3)
Adding metallic sodium at room temperature(0.6g, 0.025mmol) was added to ethylene glycol 1(5g, 0.083mmol) and heated at 80 ℃. Next, 1-bromopentane 2(3.7g, 0.025mmol) was added to the reaction mixture at the same temperature and stirring of the reaction mixture was continued for 4 hours. The mixture was filtered to remove inorganic material and diluted with ice water (200mL) and extracted with ethyl acetate (2X 200 mL). The organic layer was washed with brine solution and dried (Na)2SO4) And concentrated to give pure 2- (pentyloxy) ethan-1-ol 3(2.2g, 0.0186mmol, 55%) as a yellow liquid. TLC system: 10% methanol in dichloromethane, Rf：0.5
2- (2- (pentyloxy) ethoxy) acetic acid (side chain-22)
To a stirred solution of 2- (pentyloxy) ethan-1-ol 3(500mg, 4.23mmol) in 1, 4-dioxane (10mL) was added NaOH (0.84g, 21.12mmol) at room temperature. The mixture was stirred at room temperature for 15 minutes. Tert-butyl 2-bromoacetate 4(1.3mL, 8.47mmol) and 18-crown ether (25mg) were then added to the reaction mixture and stirred at room temperature for 18 h. After the starting material was consumed, the mixture was diluted with ice-cold water (50mL) and extracted with ether (2X 50 mL). The separated aqueous layer was acidified with concentrated HCl (pH about 2) and extracted with a 10% methanol mixture in dichloromethane (2X 100 mL). The organic layer was washed with brine solution (50mL), dried over sodium sulfate and concentrated to give pure 2- (2- (pentyloxy) ethoxy) acetic acid side chain-22 (200mg, 1.05mmol, 24% yield) as a thick brown liquid. TLC system: 10% methanol in dichloromethane, R f：0.10
Side chain-23
2- (2- (pentyloxy) ethoxy) ethan-1-ol (3)
To a stirred solution of methyl 2,2' -oxybis (ethan-1-ol) 1(10g, 66.25mmol) in 50% aqueous NaOH (250ml) was added 1-bromopentane 2 and stirred at 100 ℃ for 16 h. After completion of the reaction as indicated by TLC, water was added to the reaction mixture (200mL) and extracted with ethyl acetate (2 × 500 mL). The combined organic layers were washed with brine solution (200mL) and Na2SO4Drying and concentration under reduced pressure gave 2- (2- (pentyloxy) ethoxy) ethan-1-ol 3(8.4g, 47.72mmol, 72%) as a brown liquid. TLC system: 100% ethyl acetate, Rf：0.50
4-Methylbenzenesulfonic acid 2- (2- (pentyloxy) ethoxy) ethyl ester (4)
To a stirred solution of 2- (2- (pentyloxy) ethoxy) ethan-1-ol 3(4g, 22.72mmol) in THF (30mL) at 0 deg.C was added a solution of NaOH (1.8g, 45.45mmol) in water (10 mL). The reaction mixture was stirred at room temperature for 15 minutes, then cooled to 0 ℃, p-toluenesulfonyl chloride (5.1g, 27.27mmol) was added to the reaction mixture and stirred at room temperature for 16 hours. After the starting material was consumed, the mixture was diluted with ethyl acetate (150mL) and washed with ice-cold water (2 × 30mL), brine solution (30mL), dried over sodium sulfate and concentrated under reduced pressure. The crude compound was purified by 100-mesh 200-mesh silica gel column chromatography eluting with 30% ethyl acetate in petroleum ether to give 2- (2- (pentyloxy) ethoxy) ethyl 4-methylbenzenesulfonate 4(5.4g, 16.36mmol, 72% yield) as a brown liquid. TLC system: 70% ethyl acetate in petroleum ether, R f：0.50；LCMS：m/z＝331.09(M+H)+
1- (2- (2-azidoethoxy) ethoxy) pentane (5)
To 4-methylbenzenesulfonic acid at room temperatureTo a stirred solution of acid 2- (2- (pentyloxy) ethoxy) ethyl ester 4(2g, 6.06mmol) in DMF (20mL) was added NaN3(630mg, 9.696mmol) and then stirred at 60 ℃ for 16 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, ice water (2 × 50mL) was added to the crude material and extracted with ether (2 × 100 mL). The combined organic layers were washed with brine solution (50mL) and Na2SO4Dried and concentrated under reduced pressure to give 1- (2- (2-azidoethoxy) ethoxy) pentane 5(1.1g, 5.97mmol, 91%) as a brown liquid. TLC system: 50% ethyl acetate in hexane, Rf：0.30；LCMS：m/z＝202(M+H)+
2- (2- (pentyloxy) ethoxy) ethan-1-amine (side chain-23)
To 1- (2- (2-azidoethoxy) ethoxy) pentane 5(1.1g, 5.47mmol) in THF: h2P (Me) in THF to a stirred solution of O (4:1) (10mL)3(1M, 11mL, 10.94mmol) and stirred at room temperature for 16 h. After completion of the reaction as indicated by TLC, the organic solvent was evaporated to give 2- (2- (pentyloxy) ethoxy) ethan-1-amine side-chain-23 (1.1g, 6.285mmol, crude material) as a brown liquid. TLC system: 10% methanol in dichloromethane, Rf：0.1
Side chain-24
N-methyl-2- (2- (pentyloxy) ethoxy) ethan-1-amine (side chain-24)
4-Methylbenzenesulfonic acid 2- (2- (pentyloxy) ethoxy) ethyl ester 1 (the synthesis of compound 1 is reported in side chain-23) (500mg, 1.51mmol) was added to methylamine in ethanol (1M, 5mL) at room temperature, followed by stirring at 80 ℃ for 12 hours. After completion of the reaction as indicated by TLC, the reaction mixture was evaporated, diluted with water (50mL), acidified with 1N HCl (pH ca. 2), and washed with diethyl ether (2 × 50 mL). Followed bySaturated NaHCO3The separated aqueous layer was basified with solution and extracted with 10% methanol in dichloromethane (2X 100mL), washed with brine solution (50mL), Na2SO4Dried and concentrated to give N-methyl-2- (2- (pentyloxy) ethoxy) ethan-1-amine side chain-24 (240mg, 1.27mmol, 84%) as a brown oily liquid. TLC system: methanol in dichloromethane (1 drop Et) 10%3N)，Rf：0.20
Side chain-25
N-benzyl-2- (2- (pentyloxy) ethoxy) ethan-1-amine (side chain 25)
To a stirred solution of 4-methylbenzenesulfonic acid 2- (2- (pentyloxy) ethoxy) ethyl ester 1 (the synthesis of compound 1 is reported in side chain-23) (500g, 1.515mmol) in DMF (5ml) at room temperature were added potassium carbonate (2g, 15.15mmol) and benzylamine (256mg, 2.424mmol), followed by stirring at 80 ℃ for 16 h. After completion of the reaction as indicated by TLC, ice water (30mL) was added to the reaction mixture and extracted with ethyl acetate (2 × 30 mL). The organic layer was washed with brine solution (30mL) and Na 2SO4Dried and concentrated under reduced pressure. The crude compound was purified by combinatorial flash chromatography eluting with 10% methanol in dichloromethane to afford N-benzyl-2- (2- (pentyloxy) ethoxy) ethan-1-amine side chain 25(300mg, 1.132mmol, 74.8% yield) as an off-white solid. TLC system: 10% methanol in dichloromethane, Rf：0.20；LCMS：m/z＝266.43(M+H)+
Side chain-26
4-Methylbenzenesulfonic acid undecyl ester (2)
To a stirred solution of undecan-1-ol 1(2.0g, 11.6mmol) in THF (20mL) at 0 deg.C was added NaOH (0.93g, 23.2 mmol). The mixture was stirred at room temperature for 15 minutes. Then cooled to 0 ℃, p-toluenesulfonyl chloride (2.65g, 13.92mmol) was added to the reaction mixture and stirred at room temperature for 16 hours. After the starting material was consumed, the mixture was diluted with ethyl acetate (150mL) and washed with ice-cold water (2 × 20mL), brine solution (20mL), dried over sodium sulfate and concentrated. The crude compound was purified by 100-mesh 200-mesh silica gel column chromatography eluting with 20% ethyl acetate in hexane to give undecyl 4-methylbenzenesulfonate 2 as an off-white solid (2.6g, 7.97mmol, 68% yield). TLC system: 10% methanol in dichloromethane, Rf：0.70；LCMS：m/z＝325.56(M-H)-
N-methylundecane-1-amine (side chain-26)
To a stirred solution of undecyl 4-methylbenzenesulfonate 2(600mg, 1.84mmol) in methanol (2mL) in a sealed tube at room temperature was added 1M MeNH in methanol 2(2.5mL), followed by stirring at 80 ℃ for 16 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated. Water (20mL) was added to the crude product and extracted with ethyl acetate (3X 30 mL). With 1N HCl solution (20mL) and saturated NaHCO3The organic layer was washed with Na, a solution (20mL), brine solution (100mL)2SO4Drying and concentration gave N-methylundecan-1-amine (side chain-26) (190mg, 1.02mmol, crude material) as a brown oily liquid. TLC system: 10% methanol in dichloromethane, Rf：0.20；LCMS：m/z＝186.40(M+H)+
Side chain-27
N-benzyl undecane-1-amine (side chain-27)
To a stirred solution of 4-methylbenzenesulfonic acid undecyl ester 1 (the synthesis of compound 1 is reported in side chain-26) (600mg, 1.84mmol) in ethanol (10mL) in a sealed tube was added benzylamine (0.6mL) at room temperature, followed by stirring at 60 ℃ for 16 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, water (20mL) was added to the crude material and extracted with ethyl acetate (3 × 30 mL). With 1N HCl solution (20mL) and saturated NaHCO3The organic layer was washed with Na, a solution (20mL), brine solution (50mL)2SO4Dried and concentrated to give crude material. The crude compound was purified by 100-mesh 200-mesh silica gel column chromatography eluting with 40% ethyl acetate in hexane to give N-methylundec-1-amine side chain-27 as an off-white semi-solid (350mg, 1.34mmol, 72% yield). TLC system: 50% ethyl acetate in petroleum ether, R f：0.20；LCMS：m/z＝262.47(M+H)+
Side chain-28
(3- (ethoxymethyl) phenyl) methanol (2)
A stirred solution of 1, 3-phenylenedimethanol (1) (5g, 36.496mmol) in THF (100mL) was treated with NaH (1.17g, 29.197mmol) at 0 deg.C to room temperature for 30 minutes. Iodoethane (2.3mL, 29.197mmol) in THF (10mL) was added to the above reaction mixture at ℃ and stirred at 60 ℃ under nitrogen atmosphere for 6 hours. The reaction mixture was quenched with ice water and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine (2X 100mL) and Na2SO4Drying and evaporating under reduced pressure. The crude residue was purified by combined flash chromatography using 20% ethyl acetate in petroleum ether to give (3- (ethoxymethyl) phenyl) methanol 3 as a light yellow oily liquid (2.1g, 12.65mmol, 34% yield). TLC system: 40% ethyl acetate in petroleum ether, Rf：0.50；LCMS：m/z＝120.99(M-46)+
2- (3- (ethoxymethyl) benzyloxy) acetic acid tert-butyl ester (4)
To a stirred solution of (3- (ethoxymethyl) phenyl) methanol 2(750mg, 4.158mmol) in 1, 4-dioxane (10mL) was added NaOH (542mg, 13.55mmol) at 0 ℃. The mixture was stirred at room temperature for 15 minutes, then cooled to 0 ℃, tert-butyl 2-bromoacetate 3(1.76g, 9.036mmol) and 18-crown ether (40mg) were added, and stirred at room temperature for 16 hours. After the starting material was consumed, the reaction mixture was diluted with ethyl acetate (50mL) and washed with ice-cold water (50mL), brine solution (20mL), dried over sodium sulfate and concentrated under reduced pressure. The crude compound was purified by combined flash chromatography and eluted with 10% ethyl acetate in petroleum ether to give tert-butyl 2- (3- (ethoxymethyl) benzyloxy) acetate 4(900mg, 3.21mmol, 71.1% yield) as a colorless oily liquid. TLC system: 30% ethyl acetate in petroleum ether, R f：0.50
2- (3- (ethoxymethyl) benzyloxy) acetic acid (side chain-28)
To a stirred solution of tert-butyl 2- (3- (ethoxymethyl) benzyloxy) acetate 4(300mg, 1.0714mmol) in 1, 4-dioxane (2mL) in a sealed tube was added 4M HCl in 1, 4-dioxane (2.5mL) at room temperature, followed by stirring at reflux for 4 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated, water (50mL) was added to the crude material and extracted with ethyl acetate (2 × 30 mL). The organic layer was washed with brine solution (30mL) and Na2SO4Dried and concentrated under reduced pressure. The crude compound was purified by wet milling with diethyl ether (30mL) and side chain-28 of 2- (3- (ethoxymethyl) benzyloxy) acetic acid was obtained as a pale yellow oily liquid (190mg, 0.848mmol, 79.1% yield). TLC system: 5% inMethanol in dichloromethane, Rf：0.20；LCMS：m/z＝225.41(M+H)+
Side chain-29
2- (3- (ethoxymethyl) benzyloxy) ethanol (2)
To a stirred solution of tert-butyl 2- (3- (ethoxymethyl) benzyloxy) acetate 1 (the synthesis of compound 1 is reported in side chain-28) (500mg, 1.7857mmol) in THF (10mL) at 0 deg.C was added 1M LiAlH in THF4(5.3mL, 5.357mmol) for 3 hours. After completion of the reaction as indicated by TLC, saturated NH was used4The reaction mixture was quenched with Cl solution (20mL) and brine solution (20mL) and extracted with ethyl acetate (2X 50 mL). The organic layer was washed with brine solution (50mL) over Na 2SO4Drying and concentration gave 2- (3- (ethoxymethyl) benzyloxy) ethanol 2(350mg, 1.666mmol, 93%) as a colorless oily liquid. TLC system: 40% ethyl acetate in hexane, Rf：0.20；LCMS：m/z＝233.33(M+Na)+
4-Methylbenzenesulfonic acid 2- (3- (ethoxymethyl) benzyloxy) ethyl ester (3)
To a stirred solution of 2- (3- (ethoxymethyl) benzyloxy) ethanol 2(1.1g, 5.23mmol) in THF (30mL) at 0 deg.C was added NaOH (0.42g, 10.47 mmol). The mixture was stirred at room temperature for 15 minutes. Then cooled to 0 ℃, p-toluenesulfonyl chloride (1.2g, 6.28mmol) was added to the reaction mixture and stirred at room temperature for 16 hours. After the starting material was consumed, the mixture was diluted with ethyl acetate (100mL) and washed with ice-cold water (2X 50mL), brine solution (50mL),dried over sodium sulfate and concentrated. The crude compound was purified by 100-mesh 200-mesh silica gel column chromatography and eluted with 10% ethyl acetate in hexane to give 2- (3- (ethoxymethyl) benzyloxy) ethyl 4-methylbenzenesulfonate 3(1.6g, 4.39mmol, 84% yield) as a colorless oily liquid. TLC system: 30% ethyl acetate in hexane, Rf：0.70；LCMS：m/z＝386.98(M+Na)+
1- ((2-azidoethoxy) methyl) -3- (ethoxymethyl) benzene (4)
To a stirred solution of 4-methylbenzenesulfonic acid 2- (3- (ethoxymethyl) benzyloxy) ethyl ester 3(1.7g, 4.39mmol) in DMF (30mL) at room temperature was added NaN 3(0.48g, 7.47mmol), followed by stirring at 60 ℃ for 16 hours. After completion of the reaction as indicated by TLC, water (2 × 100mL) was added to the crude material and extracted with ether (2 × 50 mL). The organic layer was washed with brine solution (50mL) over Na2SO4Dried and concentrated. The crude compound was purified by washing with n-pentane (100mL) to give 1- ((2-azidoethoxy) methyl) -3- (ethoxymethyl) benzene 4(1.1g, 4.68mmol, quantitative) as a brown gummy liquid. TLC system: 30% ethyl acetate in hexane; rf：0.70
2- (3- (ethoxymethyl) benzyloxy) ethylamine (side chain-29)
To 1- ((2-azidoethoxy) methyl) -3- (ethoxymethyl) benzene 4(600mg, 2.564mmol) in THF H2To a stirred solution in O (20mL) was added 1M P (Me) in THF3(7.7ml, 7.69mmol), followed by stirring at room temperature for 16 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated and the crude compound was co-distilled with toluene (50mL) to give 2- (3- (ethoxymethyl) benzyloxy) ethylamine side chain-29 (500mg, 2.39mmol, 93%) as a brown semi-solid.TLC system: 10% methanol in dichloromethane, Rf：0.20；LCMS：m/z＝210.32(M+H)+
Side chain-30
2- ((3- (ethoxymethyl) benzyl) oxy) -N-methylethyl-1-amine (side chain-30)
To a stirred solution of 2- (3- (ethoxymethyl) benzyloxy) ethyl 4-methylbenzenesulfonate 3 (the synthesis of compound 3 is reported in side chain-29) (500mg, 1.373mmol) in methanol (3mL) in a sealed tube at room temperature was added 1M MeNH in ethanol 2(5mL), followed by stirring at 80 ℃ for 16 hours. After completion of the reaction as indicated by TLC, the organic solvent was evaporated and the crude residue was diluted with water (50mL) and extracted with ethyl acetate (2 × 50 mL). With 1N HCl solution (20mL) and saturated NaHCO3The organic layer was washed with Na, a solution (20mL), brine solution (50mL)2SO4Drying and concentration gave 2- (3- (ethoxymethyl) benzyloxy) -N-methylethylamine side chain-30 (300mg, 1.345mmol, crude material) as a yellow solid. TLC system: 10% methanol/dichloromethane, Rf：0.20；LCMS：m/z＝224.38(M+H)+
Side chain-31
2- (3- (2-ethoxyethoxy) phenyl) acetic acid (side chain-31)
To a stirred solution of 2- (3-hydroxyphenyl) acetic acid 1(2.0g, 13.157mmol) in 10% NaOH solution (10ml) and DMSO (20ml) at room temperature was added. The mixture was stirred at room temperature for 15 minutes. 1-bromo-2-ethoxyethane 2(2.01g, 13.15mmol) was then added to the reaction mixture and stirred at 80 ℃ for 4 hours. After the starting material was consumed, the mixture was quenched with 1N HCl solution (20mL) and extracted with ethyl acetate (3 × 50mL), and washed with brine solution (50mL), dried over sodium sulfate and concentrated. The crude compound passes through (10)0-200 mesh silica gel) column chromatography and elution with 40% ethyl acetate in hexanes to afford side chain-31- (3- (2-ethoxyethoxy) phenyl) acetic acid as an off-white solid (520mg, 2.32mmol, 18%). TLC system: 100% ethyl acetate, R f：0.60；LCMS：m/z＝225.08(M+H)+
Side chain-32
3- (2-Ethoxyethoxy) benzaldehyde (3)
To a stirred solution of 3-hydroxybenzaldehyde 1(15g, 122.9mmol) in DMSO (100mL) and 10% aqueous NaOH was added 1-bromo-2-ethoxyethane 2(34mL, 307.3mmol) in DMSO (50mL) dropwise at 80 deg.C, followed by stirring at the same temperature for 10 hours. After completion of the reaction as indicated by TLC, the reaction mixture was poured into 1M HCl solution (200mL) and extracted with ether (2X 500 mL). The combined organic layers were washed with brine solution (100mL) and Na2SO4Dried and concentrated under reduced pressure to give 3- (2-ethoxyethoxy) benzaldehyde 3(5.4g, 27.83mmol, 22%) as a colorless oily liquid. TLC system: 30% ethyl acetate in petroleum ether, Rf：0.50；LCMS：m/z＝195.31(M+H)+
(E) -1- (2-ethoxyethoxy) -3- (2-nitrovinyl) benzene (4)
To a stirred solution of 3- (2-ethoxyethoxy) benzaldehyde (200mg, 1.03mmol) in nitromethane (0.55mL, 10.3mmol) was added ammonium acetate (88mg, 1.133mmol) at room temperature, followed by stirring at 100 ℃ for 12 hours. After completion of the reaction as indicated by TLC, the reaction mixture was diluted with water (50ml) and acetic acidExtraction with ethyl ester (2X 30 mL). The combined organic layers were washed with brine solution (20mL) and Na2SO4Dried and concentrated under reduced pressure. The crude compound was purified by Grace column chromatography eluting with 60% acetonitrile in 0.1% aqueous formic acid to give (E) -1- (2-ethoxyethoxy) -3- (2-nitrovinyl) benzene 4(120mg, 0.506mmol, 49%) as a colorless gum liquid. TLC system: 100% methylene chloride, R f：0.50；LCMS：m/z＝238.31(M+H)+
2- (3- (2-ethoxyethoxy) phenyl) ethan-1-amine (side chain-32)
To a stirred solution of (E) -1- (2-ethoxyethoxy) -3- (2-nitrovinyl) benzene 4(1.25g, 5.27mmol) in THF (15mL) was added LAH (1M, 26.3mL, 26.37mmol) in THF at 0 deg.C, followed by stirring at 75 deg.C for 16 h. After completion of the reaction as indicated by TLC, the reaction mixture was diluted with ice water (100mL) and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with brine solution (50mL) and Na2SO4Drying and concentration gave 2- (3- (2-ethoxyethoxy) phenyl) ethan-1-amine side chain-32 (310mg, 1.483mmol, crude material) as a reddish brown gummy liquid. TLC system: 10% methanol in dichloromethane, Rf：0.10；LCMS：m/z＝210.32(M+H)+
Side chain-33
2- (3- (2-ethoxyethoxy) phenyl) -N-methylacetamide (2)
Synthesis of 2- (3- (2-ethoxyethoxy) phenyl) acetic acid 1 (compound 1 is reported in side chain-31) at 0 deg.C (40)0mg, 1.785mmol) in DMF (5mL) was added methylamine hydrochloride (299mg, 4.464mmol), HATU (458mg, 2.678mmol) and DIPEA (1.24mL, 7.143mmol) and stirred at room temperature for 16 h. After completion of the reaction as indicated by TLC, the organic solvent was evaporated and water (50mL) was added to the crude residue and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with brine solution (30mL) and Na 2SO4Dried and concentrated under reduced pressure. The crude compound was purified by 100-phase 200 silica gel column chromatography eluting with 2% methanol in dichloromethane to give 2- (3- (2-ethoxyethoxy) phenyl) -N-methylacetamide 2(270mg, 1.139mmol, crude material) as an off-white solid. TLC system: 5% methanol in dichloromethane, Rf：0.50；LCMS：m/z＝238.28(M+H)+
2- (3- (2-ethoxyethoxy) phenyl) -N-methylethyl-1-amine (side chain-33)
To a stirred solution of 2- (3- (2-ethoxyethoxy) phenyl) -N-methylacetamide 2(165mg, 0.696mmol) in THF (2mL) at 0 deg.C was added 1M BH3THF (2.08mL, 2.088mmol), then stirred at room temperature for 4 h. After completion of the reaction as indicated by TLC, the reaction mixture was quenched with methanol (20mL) and the organic solvent was evaporated. To the crude residue was added water (50mL) and extracted with ethyl acetate (2X 50 mL). The combined organic layers were washed with brine solution (30mL) and Na2SO4Dried and concentrated under reduced pressure to give 2- (3- (2-ethoxyethoxy) phenyl) -N-methylethyl-1-amine side chain-33 (125mg, 0.560mmol, crude material) as a reddish brown liquid. TLC system: 5% methanol in dichloromethane, Rf：0.70
Side chain-34
4-Methylbenzenesulfonic acid 2- (2- (2-ethoxyethoxy) ethoxy) ethyl ester (2)
To a stirred solution of 2- (2- (2-ethoxyethoxy) ethoxy) ethan-1-ol 1(8.5g, 47.69mmol) in THF (30mL) at 0 deg.C was added NaOH (3.89g, 97.415mmol) in water (10 mL). The mixture was stirred at room temperature for 15 minutes. Then cooled to 0 ℃, p-toluenesulfonyl chloride (11.63g, 61.04mmol) was added to the reaction mixture and stirred at room temperature for 16 hours. After the starting material was consumed according to TLC, the mixture was diluted with ethyl acetate (150mL) and washed with ice-cold water (2 × 20mL), brine solution (20mL), dried over sodium sulfate and concentrated. The crude compound was purified by 100-mesh 200-mesh silica gel column chromatography eluting with 20% ethyl acetate in petroleum ether to give 2- (2- (2-ethoxyethoxy) ethoxy) ethyl 4-methylbenzenesulfonate 2(4g, 12.048mmol, 25% yield) as a brown liquid. TLC system: 40% ethyl acetate in petroleum ether, R f：0.50
N-benzyl-2- (2- (2-ethoxyethoxy) ethoxy) ethan-1-amine (side chain-34)
To a stirred solution of 4-methylbenzenesulfonic acid 2- (2- (2-ethoxyethoxy) ethoxy) ethyl ester 2(500g, 1.506mmol) in DMF (5ml) at room temperature were added potassium carbonate (2g, 15.06mmol) and benzylamine (273mg, 2.409mmol) and stirred at 80 ℃ for 16 h. After completion of the reaction (indicated by TLC), ice water was added to the reaction mixture (100mL) and extracted with ethyl acetate (2 × 100 mL). The organic layer was washed with brine solution (50mL) over Na2SO4Dried and concentrated under reduced pressure. The crude compound was purified by combinatorial flash chromatography eluting with 10% methanol in DCM to give N-benzyl-2- (2- (2-ethoxyethoxy) ethoxy) ethan-1-amine side chain 34 as a yellow liquid (500mg, 0.93mmol, 62% yield). An LC system: 10% methanol in dichloromethane, Rf：0.20
Side chain-35
5- (Pentayloxy) pentanoic acid ethyl ester (3)
To a stirred solution of pentan-1-ol 1(1.0g, 11.36mmol) in N, N-dimethylformamide (10mL) at 0 deg.C was added NaH (450mg, 11.63mmol) and stirred at room temperature for 30 min. Ethyl 5-bromovalerate 2(1.99mL, 12.49mmol) in N, N-dimethylformamide (5mL) was added to the above reaction mixture at 0 ℃ and stirred at room temperature under a nitrogen atmosphere for 16 hours. The reaction mixture was quenched with aqueous ammonium chloride solution and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with brine (2X 50mL) and Na 2SO4Drying and evaporating under reduced pressure. The crude residue was purified by column chromatography using 2% ethyl acetate in hexane to give ethyl 5- (pentyloxy) valerate 3(200mg, 1.7187mmol, 8% yield) as a light yellow oily liquid. An LC system: 5% ethyl acetate in hexane, Rf：0.30
5- (Pentayloxy) pentanoic acid (side chain-35)
To ethyl 5- (pentyloxy) valerate (200mg, 0.92mmol) in THF H at 0 deg.C2To a stirred solution of O in a mixture (4:1, 5mL) was added lithium hydroxide (88mg, 3.073mmol) and stirred at room temperature for 16 h. The reaction mixture was evaporated under reduced pressure and the crude residue was washed with diethyl ether and acidified with 1N aqueous HCl and extracted with ethyl acetate (3 × 10 ml). The combined organic layers were washed with brine (2X 15mL) and Na2SO4Dried and concentrated. The residue was washed with pentane (2X 30mL) to give side chain-35 (150mg, 0.797 mmol) as an oily liquidYield 86%). An LC system: 50% ethyl acetate in hexane, Rf：0.10；LCMS：m/z＝189.21(M+H)+
Side chain-36
5- (pentyloxy) pent-1-ol (3)
To a stirred solution of pentane-1, 5-diol 1(10g, 96.153mmol) in N, N-dimethylformamide (70mL) at 0 deg.C was added 60% NaH (4.2g, 105.768mmol) and stirred at room temperature for 1 hour. A solution of 1-bromopentane (14.5g, 96.153mmol) in N, N-dimethylformamide (30mL) was added to the above reaction mixture at 0 deg.C and stirred at room temperature under a nitrogen atmosphere for 16 hours. The reaction mixture was quenched with ice water and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine (2X 100mL) and Na 2SO4Drying and evaporating under reduced pressure. The crude residue was purified by 100-phase 200 silica gel column chromatography using 20% ethyl acetate in hexane to give 5- (pentyloxy) pentan-1-ol 3 as a pale yellow oily liquid (7g, 40.23mmol, 43% yield). An LC system: 50% ethyl acetate in hexane, Rf：0.50
4-Methylbenzenesulfonic acid 5- (pentyloxy) pentyl ester (4)
To a stirred solution of 5- (pentyloxy) pentan-1-ol 3(4g, 22.98mmol) in DCM (40mL) at 0 deg.C were added triethylamine (7g, 68.94mmol) and DMAP (0.28g, 2.298 mmol). The mixture was stirred at room temperature for 15 minutes. Then cooled to 0 ℃, p-toluenesulfonyl chloride (6.58g, 34.48mmol) was added to the reaction mixture and stirred at room temperature for 1 hour. After the starting material is exhausted, useThe mixture was diluted with ethyl acetate (200mL) and washed with ice-cold water (2 × 200mL), brine solution (100mL), dried over sodium sulfate and concentrated under reduced pressure. The crude compound was purified by combinatorial flash chromatography eluting with 10% ethyl acetate in hexanes to give 4-methylbenzenesulfonic acid 5- (pentyloxy) pentyl ester 4(6g, 18.29mmol, 80% yield) as a colorless oily liquid. An LC system: 20% ethyl acetate in hexane, Rf：0.50
1-azido-5- (pentyloxy) pentane (5)
To a stirred solution of 4-methylbenzenesulfonic acid 5- (pentyloxy) pentyl ester 4(4.0g, 12.195mmol) in DMF (30mL) at room temperature was added NaN3(1.2g, 18.292mmol), followed by stirring at 60 ℃ for 16 h. After completion of the reaction as indicated by TLC, the organic solvent was evaporated. Water (2X 200mL) was added to the crude residue and extracted with ethyl acetate (2X 100 mL). The organic layer was washed with brine solution (100mL) over Na2SO4Dried and concentrated. The crude compound was purified by washing with pentane (3 × 25mL) to obtain 1-azido-5- (pentyloxy) pentane 5(2.3g, 11.55mmol, 95%) as an oily liquid. TLC system: 20% ethyl acetate in hexane; rf:0.80
5- (Pentyloxy) pentan-1-amine (side chain-36)
To 1-azido-5- (pentyloxy) pentane (5) (2.3g, 11.55mmol) in THF H2To a stirred solution in O (30mL) was added 1M P (Me) in THF3(23ml, 23.11mmol) followed by stirring at room temperature for 16 h. After completion of the reaction as indicated by TLC, the organic solvent was evaporated to give 5- (pentyloxy) pent-1-amine side-chain-36 (2.8g, 16.185mmol, crude material) as an oily liquid. An LC system: 50% ethyl acetate in hexane, Rf：0.10
Side chain-37
N-methyl-5- (pentyloxy) pent-1-amine (side chain-37)
To a stirred solution of 4-methylbenzenesulfonic acid 5- (pentyloxy) pentyl ester 1 (the synthesis of compound 1 is reported in side chain-36) (6g, 18.29mmol) in ethanol (20mL) in a sealed tube at room temperature was added 1M methylamine in ethanol (91mL, 91.45mmol), followed by stirring at 80 ℃ for 16 h. After completion of the reaction as indicated by TLC, the organic solvent was evaporated. To the crude residue was added water (2X 200mL) and extracted with ethyl acetate (2X 100 mL). The organic layer was washed with 1% aqueous triethylamine (200mL), brine (100mL) and Na 2SO4Drying and concentration gave N-methyl-5- (pentyloxy) pent-1-amine (side chain-37) (2.6g, 13.90mmol, 76%) as a pale yellow oily liquid. An LC system: 10% methanol in dichloromethane, Rf：0.10
Example 2: preparation of compounds
Intermediates 1 and 2
Experiment:
5-fluoro-3-iodopyridin-2-amine (2)
To 5-Fluoropyridin-2-amine (1) (10g, 89.28mmol) in 2M H at 0 deg.C2SO4To a stirred solution in aqueous solution (150mL) was added sodium metaperiodate (7.5g, 35mmol) and heated at 100 ℃. Next, sodium iodide (13.5g, 89.28mmol) in water (30mL) was added dropwise at the same temperature and maintained for 2 hours. After the starting material was consumed, the mixture was poured into ice-cold water (300mL) and solid NaHCO was used3Alkalization (pH)About 8) and extracted with ethyl acetate (2 × 500 mL). The combined organic layers were washed successively with sodium thiosulfate solution (2X 200mL), water (200mL), brine solution (100mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude compound was purified by column chromatography using 100-200 silica and eluted with 15% to 20% ethyl acetate in petroleum ether to give 5-fluoro-3-iodopyridin-2-amine (2) (12g, 50mmol, 56%) as a brown solid. TLC system: 30% EtOAc in hexane, Rf：0.5LCMS(ESI):m/z 239[M+H]+
5-fluoro-3- ((trimethylsilyl) ethynyl) pyridin-2-amine (3)
A mixture of 5-fluoro-3-iodopyridin-2-amine (12g, 50mmol), trimethylsilylacetylene (10mL, 75mmol), and triethylamine (40mL) in DMF (20mL) was degassed with argon for 15 minutes. Next, dichlorobis (triphenylphosphine) palladium (II) (350mg, 0.5mmol) and CuI (280mg, 1.5mmol) were added and the reaction mixture was heated to 50 ℃ and stirred for 2 h. After the starting material was consumed, the mixture was filtered through a pad of celite, which pad was washed thoroughly with ethyl acetate. The combined filtrates were washed with water, brine solution and dried (anhydrous Na)2SO4) And concentrated. The crude compound was purified by column chromatography using 100-200 silica and eluted with 5% to 10% ethyl acetate in petroleum ether to give 5-fluoro-3- ((trimethylsilyl) ethynyl) pyridin-2-amine (3) (8g, 38mmol, 76%) as a yellow thick liquid. TLC system: 20% EtOAc in hexane, Rf：0.4LCMS(ESI):m/z 209[M+H]+
5-fluoro-1H-pyrrolo [2,3-b ] pyridine (4)
To a stirred solution of 5-fluoro-3- ((trimethylsilyl) ethynyl) pyridin-2-amine (3) (500mg, 2.4mmol) in NMP (5mL) was added at room temperaturePotassium tert-butoxide (430mg, 3.8mmol) was added. The reaction mixture was stirred at 130 ℃ for 2 hours. After completion of the reaction as indicated by TLC, the mixture was poured into saturated aqueous sodium chloride (50mL) and extracted with ether (3X 100 mL). The combined organic layers were washed with ice-cold water (2X 50ml), dried over anhydrous sodium sulfate and concentrated to give 5-fluoro-1H-pyrrolo [2,3-b ] as a light brown solid ]Pyridine (4) (200mg, 1.47mmol, 61%). TLC system: 20% EtOAc in hexane, Rf：0.3LCMS(ESI):m/z 137[M+H]+。
3-bromo-5-fluoro-1H-pyrrolo [2,3-b ] pyridine (5)
At 0 deg.C to 5-fluoro-1H-pyrrolo [2,3-b ]]NBS (6.4g, 36mmol) was added portionwise to a stirred solution of pyridine (4.5g, 33mmol) in DMF (50mL) and allowed to stand for 2 hours. After completion of the reaction as indicated by TLC, the reaction mixture was poured into ice-cold water (100mL), the precipitated solid was filtered and dried in vacuo to give 3-bromo-5-fluoro-1H-pyrrolo [2,3-b ] as a light brown solid]Pyridine (5) (4.6g, 21mmol, 64%). TLC system: 20% EtOAc in hexane, Rf：0.5LCMS(ESI):m/z 215[M+H]+
3-bromo-5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] pyridine (6)
To a stirred suspension of NaH (1.3g, 34mmol) in DMF (30mL) at 0 deg.C was added 3-bromo-5-fluoro 1H pyrrolo [2,3-b ] in DMF]Pyridine 5(4.6g, 21 mmol). After 1h, a solution of p-TsCl (5.7g, 30mmol) in DMF (20mL) was added slowly at the same temperature and stirred for 2 h. After completion of the reaction as indicated by TLC, the mixture was poured into ice-cold water (200mL) and the precipitated solid was filtered and dried to give 3-bromo-5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] as an off-white solid]Pyridine (6) (6.2g, 16mmol, 76%). TLC system: etoa in hexane 10% c，Rf：0.8LCMS(ESI):m/z 369[M+H]+
5-fluoro-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1-tosyl-1H-pyrrolo [2,3-b ] pyridine intermediate 1
To 3-bromo-5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] at room temperature]To an argon purged solution of pyridine 6(1g, 2.7mmol) in dioxane (10mL) were added bis-pinacolato diboron (2g, 8.1mmol), potassium acetate (0.8g, 8.1mmol) and PdCl2(dppf) (0.2g, 0.27 mmol). The reaction mixture was heated at 60 ℃ for 16 hours in a sealed tube. After the starting material was consumed, the mixture was diluted with ice cold water (50mL) and extracted with EtOAc (3X 100 mL). The combined organic layers were washed with brine solution (2 × 50mL), dried over anhydrous sodium sulfate and concentrated. The crude residue was purified by combined flash chromatography to give 5-fluoro-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1-toluenesulfonyl-1H-pyrrolo [2,3-b ] as a white solid]Pyridine (intermediate 1) (620mg, 1.49mmol, 55% yield). TLC system: 20% EtOAc in hexane, Rf：0.3LCMS(ESI):m/z 417[M+H]+。
Trifluoro (5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] pyridin-3-yl) potassium borate (intermediate 2)
To 5-fluoro-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1-toluenesulfonyl-1H-pyrrolo [2,3-b ] at room temperature ]To a stirred solution of pyridine (intermediate 1) (36g, 0.086mol) in MeOH (100mL) was added 4.5M KHF2(29g, 0.389mol), and the mixture was stirred for 6 hours. After the starting material was consumed, the reaction mixture was concentrated under reduced pressure and co-distilled with MeOH 3 to 4 times. The crude compound was dissolved in acetone (200mL) and filtered to remove undissolved inorganic solids. The filtrate was concentrated under reduced pressure and wetted with diethyl etherThe resulting crude compound was triturated until the less polar spot on TLC disappeared to give trifluoro (5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] as a brown solid]Pyridin-3-yl) potassium borate (intermediate 2). TLC system: 10% MeOH in DCM, Rf：0.1LCMS(ESI):m/z 397[M+H]+。
Intermediate 12
(1S,3R) -N1- (2-chloro-5-fluoropyrimidin-4-yl) cyclohexane-1, 3-diamine (3)
To a stirred solution of (1R,3S) -cyclohexane-1, 3-diamine (1) (5g, 43mmol) in DMF (40mL) at 0 deg.C was added dropwise DIPEA (4mL, 24mmol) and 2, 4-dichloro-5-fluoropyrimidine (2) (3.8g, 23mmol) in DMF (10 mL). The reaction mixture was stirred at the same temperature for 15 minutes. After consumption of the starting material according to TLC (no compound 2 present), the mixture was diluted with ethyl acetate (200mL) and washed with ice-cold water (3 × 150mL), brine solution (50mL), dried over anhydrous sodium sulfate and concentrated. The crude compound was purified by column chromatography using 100-200 silica gel and 10% in DCM and Et 3MeOH in N eluted to give (1S,3R) -N1- (2-chloro-5-fluoropyrimidin-4-yl) cyclohexane-1, 3-diamine (3) (4g, 16.3mmol, 71% yield) as a white solid. TLC system: 10% in DCM (2 drops of Et)3MeOH in N), Rf: 0.1LCMS (ESI) M/z 245.2(M + H)+
(1S,3R) -N1- (5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] pyridin-3-yl) pyrimidin-4-yl) cyclohexane-1, 3-diamine (intermediate 12)
Degassing of (1S,3R) -N1- (2-chloro-5-fluoro-N with argonA mixture of pyrimidin-4-yl) cyclohexane-1, 3-diamine (3) (3g, 12.3mmol), intermediate 2(7.3g, 18.4mmol) and aqueous sodium carbonate (2M, 4mL) in 1,2-DME (30mL) for 15 minutes. Next, tetrakis (triphenylphosphine) palladium (0.7g, 0.6mmol) was added and the mixture was stirred at 100 ℃ for 16 h. After the starting material was consumed, the mixture was concentrated under reduced pressure. The crude compound was purified by column chromatography using 100-200 silica and 10% in DCM and Et3MeOH in N eluted the compound to give (1S,3R) -N1- (5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b) as a brown solid]Pyridin-3-yl) pyrimidin-4-yl) cyclohexane-1, 3-diamine (intermediate 12) (3.5g, 7.02mmol, 58%). TLC system: 10% in DCM (2 drops of Et)3MeOH in N), Rf: 0.2LCMS (ESI) M/z 498.98(M + H) +
Intermediate 13
(R) -3- ((2-chloro-5-fluoropyrimidin-4-yl) amino) piperidine-1-carboxylic acid tert-butyl ester
To a stirred solution of (R) -3-aminopiperidine-1-carboxylic acid tert-butyl ester (1) (1.437g, 7.18mmol) in DMF (10mL) at 0 deg.C was added dropwise DIPEA (1.56mL, 8.98mmol) and 2, 4-dichloro-5-fluoropyrimidine (2) (1.0g, 5.98mmol) in DMF (10 mL). The reaction mixture was stirred at room temperature for 16 hours. After consumption of the starting material according to TLC (no compound 2 present), the mixture was diluted with ethyl acetate (50mL) and washed with ice-cold water (3 × 10mL), brine solution (10mL), dried over anhydrous sodium sulfate and concentrated. The crude compound was purified by column chromatography using 100-200 silica and eluted with 20% EA in petroleum ether to give tert-butyl (R) -3- ((2-chloro-5-fluoropyrimidin-4-yl) amino) piperidine-1-carboxylate (3) (1.6g, 4.84mmol, 81% yield) as an off-white solid. TLC system: EA 30% in petroleum ether, Rf: 0.5LCMS (ESI) M/z 330.74(M + H)+
(R) -3- ((5-fluoro-2- (5-fluoro-1-toluenesulfonyl-1H-pyrrolo [2,3-b ] pyridin-3-yl) pyrimidin-4-yl) amino) piperidine-1-carboxylic acid tert-butyl ester
A mixture of tert-butyl (R) -3- ((2-chloro-5-fluoropyrimidin-4-yl) amino) piperidine-1-carboxylate (3) (600mg, 1.81mmol), intermediate Int-2(1.0g, 2.54mmol) and aqueous sodium carbonate (2M, 3mL) in 1,2-DME (12mL) was degassed with argon for 15 min. Then tetrakis (triphenylphosphine) palladium (104mg, 0.09mmol) was added and degassed again with argon for 10 min. The reaction mixture was stirred at 90 ℃ for 16 hours. After the starting material was consumed, the mixture was concentrated under reduced pressure. The crude compound was purified by column chromatography using 100-200 silica, and the compound was eluted with 30% ethyl acetate-petroleum ether to give (R) -3- ((5-fluoro-2- (5-fluoro-1-toluenesulfonyl-1H-pyrrolo [2, 3-b) as an off-white solid ]Pyridin-3-yl) pyrimidin-4-yl) amino) piperidine-1-carboxylic acid tert-butyl ester 4(750mg, 1.28mmol, 71% yield). TLC system: EA 30% in petroleum ether, Rf: 0.5LCMS (ESI): M/z 585.12(M + H)+
(R) -5-fluoro-2- (5-fluoro-1H-pyrrolo [2,3-b ] pyridin-3-yl) -N- (piperidin-3-yl) pyrimidin-4-amine hydrochloride
Placing (R) -3- ((5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b) in a sealed tube]Pyridin-3-yl) pyrimidin-4-yl) amino) piperidine-1-carboxylic acid tert-butyl ester (4) (300mg, 0.51mmol), acetonitrile (4mL) and 4N HCl in dioxane (2 mL). Followed by heating at 70 ℃ for 16 hours. After the starting material was consumed, the mixture was wet-milled with ether to form a white precipitate, which was filtered and further wet-milled with ether to give (R) -5-fluoro-2- (5-fluoro-1H-pyrrolo [2,3-b ] as an off-white solid]Pyridin-3-yl) -N- (piperidin-3-yl) pyrimidin-4-amine hydrochloride (intermediate 13) (147mg, 0.40mmol, 78% yield). TLC system: 10% MeOH-DCM Rf: 0.3LCMS(ESI):m/z 331.37(M+1)+
Intermediate 14
(S) -3- ((2-chloro-5-fluoropyrimidin-4-yl) amino) piperidine-1-carboxylic acid tert-butyl ester
To a stirred solution of (S) -3-aminopiperidine-1-carboxylic acid tert-butyl ester (1) (1.437g, 7.18mmol) in DMF (10mL) at 0 deg.C was added dropwise DIPEA (1.56mL, 8.98mmol) and 2, 4-dichloro-5-fluoropyrimidine (2) (1.0g, 5.98mmol) in DMF (2 mL). The reaction mixture was stirred at room temperature for 16 hours. After consumption of the starting material according to TLC (no compound 2 present), the mixture was diluted with ether (75mL) and washed with ice-cold water (3 × 50mL), brine solution (25mL), dried over sodium sulfate and concentrated under reduced pressure to give tert-butyl (S) -3- ((2-chloro-5-fluoropyrimidin-4-yl) amino) piperidine-1-carboxylate (3) (1.670g, 5.06mmol, 84% yield) as a white solid. TLC system: EA 30% in petroleum ether, Rf: 0.5LCMS (ESI) M/z 331.44(M +1) +
(S) -3- ((5-fluoro-2- (5-fluoro-1-toluenesulfonyl-1H-pyrrolo [2,3-b ] pyridin-3-yl) pyrimidin-4-yl) amino) piperidine-1-carboxylic acid tert-butyl ester
A mixture of (S) -3- ((2-chloro-5-fluoropyrimidin-4-yl) amino) piperidine-1-carboxylate (3) (2.5g, 7.57mmol), intermediate 2(4.49g, 11.36mmol) and aqueous sodium carbonate (2M, 10mL) in 1,2-DME (40mL) was degassed with argon for 15 minutes. Tetrakis (triphenylphosphine) palladium (0.437g, 0.37mmol) was added and degassed again with argon for 10 min. The reaction mixture was heated at 90 ℃ for 16 hours. After the starting material was consumed, the mixture was concentrated under reduced pressure. Crude compound passing columnChromatography using 100-200 silica and elution of the compound with 30% ethyl acetate-petroleum ether afforded (S) -3- ((5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b) as a brown solid]Pyridin-3-yl) pyrimidin-4-yl) amino) piperidine-1-carboxylic acid tert-butyl ester 4(3.650g, 6.25mmol, 82% yield). TLC system: 40% EtOAc in petroleum ether, Rf: 0.5LCMS (ESI) M/z 585.48(M + H)+
(S) -5-fluoro-2- (5-fluoro-1H-pyrrolo [2,3-b ] pyridin-3-yl) -N- (piperidin-3-yl) pyrimidin-4-amine hydrochloride
(S) -3- ((5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b) in acetonitrile (40mL) was added in a sealed tube ]Pyridin-3-yl) pyrimidin-4-yl) amino) piperidine-1-carboxylic acid tert-butyl ester (4) (3.5g, 5.99mmol) and 4N HCl in dioxane (20 mL). Followed by heating at 70 ℃ for 16 hours, after depletion of the starting material, wet milling the mixture with diethyl ether, filtering the precipitated white solid and further wet milling with diethyl ether to give (S) -5-fluoro-2- (5-fluoro-1H-pyrrolo [2,3-b ] as an off-white solid]Pyridin-3-yl) -N- (piperidin-3-yl) pyrimidin-4-amine hydrochloride (intermediate 14) (2.020g, 5.51mmol, 92% yield). TLC system: 10% MeOH-DCM Rf: 0.3LCMS (ESI) M/z 331.02(M + H)+
Intermediate 18
3-bromo-5-chloro-1H-pyrrolo [2,3-b ] pyridine (2)
To 5-chloro-1H-pyrrolo [2,3-b ] at 0 DEG C]NBS (7.02g, 39.46mmol) was added portionwise to a stirred solution of pyridine 1(5.0g, 32.89mmol) in acetone (50mL) and stirred for 2 hours. Such as TLCAfter completion of the reaction was indicated, the reaction mixture was concentrated in vacuo. The crude material was diluted with water (50mL) and extracted with EtOAc (3X 100 mL). The combined organic layers were washed with brine solution (2X 50mL), dried over anhydrous sodium sulfate and concentrated to give 3-bromo-5-chloro-1H-pyrrolo [2,3-b ] as a pale brown solid]Pyridine 2(7.0g, 30.43mmol, 92%). TLC system: 20% EtOAc in hexane, R f：0.5LCMS(ESI):m/z 230.8[M+H]+
3-bromo-5-chloro-1-tosyl-1H-pyrrolo [2,3-b ] pyridine (3)
To a stirred suspension of NaH (1.4g, 58.33mmol) in DMF (30mL) at 0 deg.C was added 3-bromo-5-chloro 1H-pyrrolo [2,3-b ] in DMF]Pyridine 2(7.0g, 30.43 mmol). After 1h, a solution of p-TsCl (6.3g, 33.47mmol) in DMF (20mL) was added slowly at the same temperature and stirred for 2 h. After completion of the reaction (as indicated by TLC), the mixture was poured into ice-cold water (200mL), and the precipitated solid was filtered and dried to give 3-bromo-5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] as an off-white solid]Pyridine 3(8.5g, 22.14mmol, 73%). TLC system: 10% EtOAc in hexane, Rf：0.8LCMS(ESI):m/z 386.4[M+H]+
5-chloro-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1-toluenesulfonyl-1H-pyrrolo [2,3-b ] pyridine (4)
To 3-bromo-5-chloro-1-tosyl-1H-pyrrolo [2,3-b ] at room temperature]To an argon purged solution of pyridine 3(2.8g, 7.29mmol) in DMF (10mL) was added bis-pinacolato diboron (3.71g, 14.58mmol), potassium acetate (2.14g, 21.87mmol) and PdCl2(dppf) (678mg, 0.729 mmol). The reaction mixture was heated at 60 ℃ for 16 hours in a sealed tube. After the starting material was consumed, the mixture was diluted with ice cold water (50mL) and with EtOAc (3 × 1) 00mL) was extracted. The combined organic layers were washed with brine solution (2 × 50mL), dried over anhydrous sodium sulfate and concentrated. The crude residue was purified by combined flash chromatography to give 5-chloro-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1-toluenesulfonyl-1H-pyrrolo [2,3-b ] as a white solid]Pyridine 4(602mg, 1.39mmol, 19% yield). TLC system: 20% EtOAc in hexane, Rf：0.3LCMS(ESI):m/z 432.7[M+H]+。
Potassium (5-chloro-1-tosyl-1H-pyrrolo [2,3-b ] pyridin-3-yl) trifluoroborate (intermediate 18)
To 5-chloro-3- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1-toluenesulfonyl-1H-pyrrolo [2,3-b ] at room temperature]To a stirred solution of pyridine 4(9.0g, 20.83mmol) in MeOH (50mL) was added 4.5M KHF2(7.32g, 93.74mmol) for 6 hours. After the starting material was consumed, the reaction mixture was concentrated under reduced pressure and co-distilled with MeOH 3 to 4 times. The crude compound was dissolved in acetone (100mL) and the undissolved inorganic solid was filtered. The filtrate was concentrated under reduced pressure to give the crude compound, which was wet-milled with ether until the less polar spot on TLC disappeared. Followed by filtration and drying of the solid to obtain trifluoro (5-chloro-1-tosyl-1H-pyrrolo [2, 3-b) as a brown solid ]Pyridin-3-yl) potassium borate intermediate 18(3.2g, 7.766mmol, 37%). TLC system: 10% MeOH in DCM; rf：0.1LCMS(ESI):m/z 351[M-60]+。
Intermediate 19
(1R,4R) -bicyclo [2.2.2] oct-5-ene-2-carboxylic acid ethyl ester (2)
Cyclohexane-1, 3-diene 1(15.0g, 187.19mmol), ethyl acrylate (18.74g, 187.19mmol) and methylene blue (119mg, 0.37mmol) were placed in a sealed tube and the mixture was heated at 140 ℃ for 16 h. After exhaustion of the starting material, (1R,4R) -bicyclo [2.2.2] is obtained as a colorless oil by fractional distillation (at 70 ℃ C. and 0.1 mmHg)]Oct-5-ene-2-carboxylic acid ethyl ester 2(20.0g, 111.11mmol, 59% yield). 1HNMR coincided. TLC system: 5% EA, R in petroleum etherf：0.5
(1S,4S) -bicyclo [2.2.2] octane-2-carboxylic acid ethyl ester (3)
In a Parr shaker, to (1R,4R) -bicyclo [2.2.2]To a solution of octyl-5-ene-2-carboxylic acid ethyl ester 2(20.0g, 111.11mmol) in EtOH (100mL) was added 10% Pd/C (2.0g, 10% w/w). At H2The reaction mixture was stirred under gas at 50Psi for 16 hours. After the starting material was consumed, the mixture was filtered through a pad of celite, which pad was washed thoroughly with ethanol. The combined filtrates were concentrated under reduced pressure to give (1S,4S) -bicyclo [2.2.2] as a colorless oil]Octane-2-carboxylic acid ethyl ester 3(18.5g, 101.64mmol, 91% yield). 1HNMR coincidence, TLC system: 5% EtOAc in petroleum ether, Rf: 0.5
(1S,4S) -bicyclo [2.2.2] oct-2-ene-2-carboxylic acid ethyl ester (4)
To a stirred solution of freshly distilled DIPA (7.68mL, 54.94mmol) in THF (50mL) at 0 deg.C was added a solution of 2.5M n-BuLi (in hexane, 19.78mL, 49.45mmol) and stirred for 30 min. (1S,4S) -bicyclo [2.2.2] is added at-78 DEG C]A solution of octane-2-carboxylic acid ethyl ester 3(5.0g, 27.47mmol) in THF (10mL) was stirred at the same temperature for 1 hour. A solution of PhSeBr (9.724g, 41.20mmol) in THF was then added at-78 deg.C and allowed to warm to 0 deg.C and stir for 30 minutes. Followed by sequential addition of H at 0 deg.C2O(35mL)、H2O2(20mL), AcOH (7.5mL), and stirred at room temperature for 1 hour, diluted with ethyl acetate and the two layers separated, and the aqueous layer extracted with ethyl acetate (2X 50 mL). The combined organic layers were washed with water, brine solution, and Na2SO4Dried and concentrated. The crude compound was purified by column chromatography using 100-200 silica and eluted with 5% to 10% ethyl acetate in petroleum ether to give (1S,4S) -bicyclo [2.2.2] as a yellow liquid]Oct-2-ene-2-carboxylic acid ethyl ester 4(4.02g, 22.33mmol, 81% yield). TLC system: 5% EtOAc in petroleum ether, Rf: 0.5LCMS (ESI) M/z 181.1(M + H)+
Intermediate 20
2-chloro-5-fluoro-4- (methylthio) pyrimidine
To a stirred solution of 2, 4-dichloro-5-fluoropyrimidine 1(5.0g, 29.94mmol) in THF (40mL) at-30 deg.C was added a 15% aqueous solution of sodium thiomethoxide (15.36mL, 32.93mmol) and stirred for 2 hours. The reaction mixture was diluted with water (50mL) and extracted with ethyl acetate (2X 150 mL). The combined organic layers were washed with brine solution (50mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 2-chloro-5-fluoro-4- (methylthio) pyrimidine 2(5.0g, 28.08mmol, 93%) as an off-white solid. TLC system: 5% EtOAc in petroleum ether, Rf: 0.6LCMS (ESI) M/z 179.08(M +1)+
5-fluoro-3- (5-fluoro-4- (methylthiomethyl) pyrimidin-2-yl) -1-tosyl-1H-pyrrolo [2,3-b ] pyridine
A mixture of 2-chloro-5-fluoro-4- (methylthio) pyrimidine 2(1g, 5.61mmol), intermediate 2(2.66mg,6.74mmol) and aqueous sodium carbonate (2M, 5mL) in 1,2-DME (25mL) was degassed with argon for 15 minutes. Then, add four(triphenylphosphine) palladium (324mg, 0.28mmol) and the mixture was stirred at 90 ℃ for 16 h. After the starting material was consumed, the mixture was poured into water (200mL), and the precipitated brown solid was filtered and dried to give the crude material 5-fluoro-3- (5-fluoro-4- (methylthio) pyrimidin-2-yl) -1-tosyl-1H-pyrrolo [2,3-b ]Pyridine 3(2g) and traces of triphenylphosphine oxide, which was used in the next step without further purification. TLC system: 20% EtOAc in petroleum ether, Rf: 0.6LCMS (ESI) M/z433.15(M +1)+
5-fluoro-3- (5-fluoro-4- (methylsulfinyl) pyrimidin-2-yl) -1-tosyl-1H-pyrrolo [2,3-b ] pyridine and 5-fluoro-3- (5-fluoro-4- (methylsulfonyl) pyrimidin-2-yl) -1-tosyl-1H-pyrrolo [2,3-b ] pyridine
5-fluoro-3- (5-fluoro-4- (methylthio) pyrimidin-2-yl) -1-tosyl-1H-pyrrolo [2,3-b ] at 0 deg.C]To a mixture of pyridine 3(300mg, 0.69mmol) in dichloromethane (15mL) was added 70% m-CPBA (188mg, 0.76mmol) portionwise for 30 min, then allowed to reach room temperature and stirred for 4 h. After the starting material was consumed, the mixture was quenched with saturated ammonium chloride solution (50mL) and extracted with ethyl acetate (2 × 50mL) and washed with brine solution (25mL), dried over sodium sulfate and concentrated under reduced pressure. The crude compound was purified by column chromatography using 100-200 silica and eluted with 20% ethyl acetate-petroleum ether to give 5-fluoro-3- (5-fluoro-4- (methylsulfonyl) pyrimidin-2-yl) -1-toluenesulfonyl-1H-pyrrolo [2,3-b ] as an off-white solid]Pyridine intermediate 20A (90mg), and elution with 80% ethyl acetate-petroleum ether, gave 5-fluoro-3- (5-fluoro-4- (methylsulfinyl) pyrimidin-2-yl) -1-tosyl-1H-pyrrolo [2,3-b ] as an off-white solid ]Pyridine intermediate 20(60 mg). Both compounds were used in the next step without further purification. TLC system for intermediate 20 a: 40% EtOAc in petroleum ether, Rf: 0.4LCMS (ESI) M/z 433.15(M +1)+(ii) a TLC system for intermediate 20: 40% EtOAc in petroleum ether, Rf: 0.1LCMS (ESI) M/z 433.15(M +1)+
Target compound
2- (2,2, 2-Trifluoroethoxy) Ethyl carbamic acid tert-butyl ester (3)
To a stirred solution of 2,2, 2-trifluoroethanol 1(3.0g, 30.00mmol) in toluene was added tert-butyl 2-hydroxyethylcarbamate 2(0.97g, 6.00mmol), 1' - (azabicycloorbonyl) dipiperidine (3.0g, 12.00mmol), tributylphosphine (3.0ml, 12.00mmol) in that order at room temperature and stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure. The crude compound was purified by column chromatography (silica gel 100-200) eluting the compound with 8% ethyl acetate in petroleum ether to give the compound tert-butyl 2- (2,2, 2-trifluoroethoxy) ethylcarbamate 3(2.0g, 8.23mmol, 27% yield). TLC system: 20% ethyl acetate/petroleum ether; rf：0.6
2- (2,2, 2-Trifluoroethoxy) ethylamine hydrochloride (4)
To a stirred solution of tert-butyl 2- (2,2, 2-trifluoroethoxy) ethylcarbamate (3) (2.0g, 8.23mmol) in dioxane was added 4M HCl in dioxane (5ml) at 0 ℃ and stirred at room temperature in a sealed tube for 16 h. The reaction mixture was concentrated under reduced pressure. The crude compound was purified by wet milling with diethyl ether (3X 25ml) to give the compound 2- (2,2, 2-trifluoroethoxy) ethylamine hydrochloride 4(1.4g, 6.17mmol, 95%). TLC system: 100% EtOAc; rf: 0.1
2- (2,2, 2-trifluoroethoxy) ethylcarbamic acid (2- ((1R,3S) -3- (5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] pyridin-3-yl) pyrimidin-4-ylamino) cyclohexylcarbamoyl) pyridin-4-yl) methyl ester (5)
To N- ((1R,3S) -3- (5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b) at room temperature]Pyridin-3-yl) pyrimidin-4-ylamino) cyclohexyl) -4- (hydroxymethyl) pyridinecarboxamide (intermediate 23) (200mg, 0.315mmol) to a stirred solution in DMF was added 1,1' -carbonyldiimidazole (130mg, 0.789mmol) and stirred at room temperature for 1 hour. To the reaction mixture was added- (2,2, 2-trifluoroethoxy) ethylamine hydrochloride 4(0.11g, 0.63mmol) at room temperature and stirred at 60 ℃ for 6 hours. Ice water (20mL) was added, filtered and dried, and the crude compound was purified by column chromatography (silica gel 100-. The compound was eluted with 50% EtOAc/petroleum ether to give (2- ((1R,3S) -3- (5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b))]Pyridin-3-yl) pyrimidin-4-ylamino) cyclohexylcarbamoyl) pyridin-4-yl) methyl 2- (2,2, 2-trifluoroethoxy) ethylcarbamate (5) (180mg, 0.224mmol, 72% yield). TLC system: 100% EtOAc; rf: 0.5LCMS (ESI) M/z 803.38(M + H) +
2- (2,2, 2-trifluoroethoxy) ethylcarbamic acid (2- ((1R,3S) -3- (5-fluoro-2- (5-fluoro-1H-pyrrolo [2,3-b ] pyridin-3-yl) pyrimidin-4-ylamino) cyclohexylcarbamoyl) pyridin-4-yl) methyl ester
2- (2,2, 2-Trifluoroethoxy) ethylcarbamic acid (2- ((1R,3S) -3- (5-fluoro-2- (5-fluoro-1H-pyrrolo [2, 3-b) of Cpd-5 in THF (5.0mL) and water (5.0mL) at 0 deg.C]Pyridin-3-yl) pyrimidin-4-ylamino) cyclohexylcarbamoyl) pyridin-4-yl) methyl ester: (180mg, 0.22mmol) to which was added lithium hydroxide (21mg, 0.89mmol) and stirred at room temperature for 48 hours. The organic solvent was distilled off and water (5mL) was added to the crude material and acidified with aqueous sodium thiosulfate solution and the filtered solid was dried under reduced pressure. The crude residue was purified by preparative HPLC to give (2- ((1R,3S) -3- (5-fluoro-2- (5-fluoro-1H-pyrrolo [2, 3) as an off-white solid-b]Pyridin-3-yl) pyrimidin-4-ylamino) cyclohexylcarbamoyl) pyridin-4-yl) methyl-2- (2,2, 2-trifluoroethoxy) ethylcarbamate (70mg, 0.108mmol, 48% yield). TLC system: 100% EtOAc; rf: 0.4; LCMS (ESI) M/z 649.14(M + H)+
4- ((4-Methoxybenzylcarbamoyloxy) methyl) picolinic acid methyl ester (3)
To a stirred solution of methyl 4- (hydroxymethyl) picolinate 1(150mg, 0.898mmol) in DMF (2mL) at room temperature was added 1,1' -carbonyldiimidazole (291mg, 1.796mmol) and stirred at room temperature for 2 h. To the reaction mixture was added (4-methoxyphenyl) methylamine (246mg, 1.796mmol) and stirred at 70 ℃ for 16 h. After completion of the reaction, ice water (30mL) was added to the mixture and extracted with ethyl acetate (3X 50 mL). The separated organic layer was washed with ice water (2 × 20mL), brine solution (20mL), dried over sodium sulfate and concentrated under reduced pressure to obtain crude methyl 4- ((4-methoxybenzylcarbamoyloxy) methyl) picolinate 3(150mg, crude). The crude compound was used directly in the next step without further purification. TLC system: 70% ethyl acetate/hexane; r f：0.3；LCMS:m/z＝331.12(M+H)+
4- ((4-Methoxybenzylcarbamoyloxy) methyl) picolinic acid (4)
To methyl 4- ((4-methoxybenzylcarbamoyloxy) methyl) picolinate 3(150mg, 0.454) in THF: H2To a stirred solution of O (2:1) in 10mL was added LiOH (32mg, 1.363mmol) and stirred at room temperature for 16 h. Evaporating the organic solvent under reduced pressure to obtainThe residue was acidified with 2N HCl and the precipitated solid was filtered and dried to give 4- ((4-methoxybenzylcarbamoyloxy) methyl) picolinic acid (4) (90mg, 0.284mmol, 63% yield) as an off white solid. TLC system: 5% methanol/dichloromethane; rf：0.1；LCMS:m/z＝317.0(M+H)+
4-Methoxybenzylcarbamic acid (2- ((1S,3R) -3- (5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] pyridin-3-yl) pyrimidin-4-ylamino) cyclohexylcarbamoyl) pyridin-4-yl) methyl ester (5)
To a stirred solution of 4- ((4-methoxybenzylcarbamoyloxy) methyl) picolinic acid (4) (50mg, 0.158mmol) in DMF (2mL) at 0 deg.C was added sequentially (1R,3S) -N1- (5-fluoro-2- (5-fluoro-1-toluenesulfonyl-1H-pyrrolo [2, 3-b)]Pyridin-3-yl) pyrimidin-4-yl) cyclohexane-1, 3-diamine (intermediate 12, enantiomer 2(Ent-2)) (94mg, 0.189mmol), HATU (120mg, 0.316mmol), DIPEA (0.087mL, 0.474mmol) and stirred at room temperature for 16 h. After completion of the reaction, the reaction mixture was diluted with ice-cold water (20mL) and the precipitated solid was filtered. This material was purified by Grace reverse phase chromatography to give 4-methoxybenzylcarbamic acid (2- ((1S,3R) -3- (5-fluoro-2- (5-fluoro-1-toluenesulfonyl-1H-pyrrolo [2, 3-b) as an off-white solid ]Pyridin-3-yl) pyrimidin-4-ylamino) cyclohexylcarbamoyl) pyridin-4-yl) methyl ester (5) (90mg, 0.113mmol, 72% yield). TLC system: 100% ethyl acetate; rf：0.6；LCMS:m/z＝797.2(M+H)+
4-Methoxybenzylcarbamic acid (2- ((1S,3R) -3- (5-fluoro-2- (5-fluoro-1H-pyrrolo [2,3-b ] pyridin-3-yl) pyrimidin-4-ylamino) cyclohexylcarbamoyl) pyridin-4-yl) methyl ester
To 4-methoxybenzylcarbamic acid (2- ((1S,3R) -3- (5-fluoro)-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b)]Pyridin-3-yl) pyrimidin-4-ylamino) cyclohexylcarbamoyl) pyridin-4-yl) methyl ester (5) (90mg, 0.113mmol) in THF H2To a stirred solution of 8mL of a mixture of O (3:1) was added LiOH (13mg, 0.565mmol) and stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with water and extracted with ethyl acetate (3X 25mL), water (20mL), brine solution (15mL) over Na2SO4Dried and concentrated to give the crude product. This crude material was purified by Graves reverse phase chromatography to give benzylcarbamic acid (2- ((1R,3S) -3- (5-fluoro-2- (5-fluoro-1H-pyrrolo [2, 3-b) as an off-white solid]Pyridin-3-yl) pyrimidin-4-ylamino) cyclohexylcarbamoyl) pyridin-4-yl) methyl ester (28mg, 0.043mmol, 38% yield). TLC system: 70% EtOAc in petroleum ether; rf: 0.30; LCMS M/z 643.0(M + H) +
Target compound
N- ((1R,3S) -3- ((5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] pyridin-3-yl) pyrimidin-4-yl) amino) cyclohexyl) -4- (hydroxymethyl) pyridinecarboxamide (intermediate 23)
To a stirred solution of 4- (hydroxymethyl) picolinic acid 1(461mg, 3.012mmol) in DMF (10mL) at 0 deg.C were added diisopropylethylamine (0.8mL, 4.518mmol) and HATU (1.14g, 3.012 mmol). After 20 min, (1S,3R) -N1- (5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] is added]Pyridin-3-yl) pyrimidin-4-yl) cyclohexane-1, 3-diamine (intermediate 12, enantiomer 2(Ent-2)) (750mg, 1.506mmol) and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was diluted with ice-cold water (100mL) and the precipitated solid was filtered to give the pure compound N- ((1R,3S) -3- ((5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b) as a brown solid]Pyridin-3-yl) pyrimidinesPyridin-4-yl) amino) cyclohexyl) -4- (hydroxymethyl) pyridinecarboxamide (intermediate 23) (950mg, 75% LCMS purity). TLC system: 10% MeOH in dichloromethane; rf: 0.6; LCMS (ESI) M/z 634.31(M + H)+
((2- (((1R,3S) -3- ((5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2,3-b ] pyridin-3-yl) pyrimidin-4-yl) amino) cyclohexyl) carbamoyl) pyridin-4-yl) methoxy) carbonyl) -D-valine methyl ester (3)
To N- ((1R,3S) -3- ((5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b) at room temperature]Pyridin-3-yl) pyrimidin-4-yl) amino) cyclohexyl) -3- (hydroxymethyl) benzamide (intermediate 23) (100mg, 0.157mmol) was added to a stirred solution in DMF 1,1' -carbonyldiimidazole (64mg, 0.394mmol) and stirred at room temperature for 2 h. Ice water (30mL) was added to the reaction mixture and filtered and dried, the crude material was dissolved in DMF, D-valine methyl ester hydrochloride 2(53mg, 0.3159mmol) was added and stirred at 70 ℃ for 3 h. After completion of the reaction, the mixture was diluted with ice water (30mL), filtered and dried. The crude compound was purified by column chromatography eluting with 50% ethyl acetate/petroleum ether to give compound (((2- (((1R,3S) -3- ((5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b))]Pyridin-3-yl) pyrimidin-4-yl) amino) cyclohexyl) carbamoyl) pyridin-4-yl) methoxy) carbonyl) -D-valine methyl ester 3(65mg, 0.082mmol, 52% yield). TLC system: 70% ethyl acetate/petroleum ether; rf: 0.6; LCMS (ESI) M/z 791.38(M + H)+
((2- (((1R,3S) -3- ((5-fluoro-2- (5-fluoro-1H-pyrrolo [2, 3-b))]Pyridin-3-yl) pyrimidin-4-yl) amino) cyclohexyl) carbamoyl) pyridin-4-yl) methoxy) carbonyl) -D-valine
To (((2- (((1R,3S) -3- ((5-fluoro-2- (5-fluoro-1-tosyl-1H-pyrrolo [2, 3-b)) at 0 deg.C]Pyridin-3-yl) pyrimidin-4-yl) amino) cyclohexanesYl) carbamoyl) pyridin-4-yl) methoxy) carbonyl) -D-valine methyl ester 3(140mg, 0.1772mmol) to a stirred solution in 1,4 dioxane (1.5mL) and water (0.5mL) was added potassium hydroxide (39mg, 0.708mmol) followed by stirring at room temperature for 24 hours. The reaction mixture was concentrated under reduced pressure to give the crude compound, which was diluted with water (5mL) and acidified with citric acid solution until pH about 6. Next, the precipitated solid was filtered and dried under reduced pressure. The crude residue was purified by preparative HPLC to give an off-white solid (((2- (((1R,3S) -3- ((5-fluoro-2- (5-fluoro-1H-pyrrolo [2, 3-b))]Pyridin-3-yl) pyrimidin-4-yl) amino) cyclohexyl) carbamoyl) pyridin-4-yl) methoxy) carbonyl) -D-valine (17mg, 0.0273mmol, 15% yield). TLC system: 10% methanol in dichloromethane/acetic acid; rf: 0.1; LCMS (ESI) M/z 623.40(M + H)+
Example 3: data on selected compounds
In vitro antiviral assays
Influenza antiviral assay:
inhibition of virus-induced cytopathic effect (CPE) and cell viability after replication of influenza A (strain A/PR/8/34, ATCC VR-95) or influenza B (cell culture adapted strain B/Lee/40, ATCC VR-1535) in MDCK cells (female Care canine kidney epithelial cells, ATCC CCL-34) was measured by XTT dye reduction (Appleyard et al, antibiotic chemotherapy (J antibiotic chemotherapy) 1 (suppl 4):49-53,1975 and Shigeta et al, antibiotic and chemotherapy (antibiotic Agents Chemotherapy) 41(7): 1423-; 1427, 1997.). MDCK cells (1X 10. mu.L per well) were inactivated with 10% heat supplemented Fetal Bovine Serum (FBS), 2mM L-glutamine, 100U/mL penicillin, 100. mu.g/mL streptomycin, 1mM sodium pyruvate, and 0.1mM NEAA in Dulbecco's Minimum Essential Medium (DMEM) at 100. mu.L per well volume 4Individual cells) were grown as monolayers in 96-well flat-bottom tissue culture plates. On the day of assay setup, cell monolayers were washed three times with DPBS. Viruses were obtained from ATCC and grown in MDCK cells for the production of virus stocks. Test compounds were diluted to assay culture at twice the desired starting concentrationIn medium (DMEM, 2mM L-glutamine, 100U/mL penicillin, 100. mu.g/mL streptomycin, 50ng/mL TPCK treated trypsin, 0.1mM NEAA and 1mM sodium pyruvate), and serially diluted. Test compounds were added in triplicate at a volume of 100 μ l/well for determining efficacy, in duplicate for determining cytotoxicity, and in single per-well concentration for colorimetric evaluation immediately prior to addition of diluted virus. Ribavirin and oseltamivir carboxylate were evaluated simultaneously as control compounds. A pre-titrated aliquot of the virus was taken from the freezer (-80 ℃) and quickly thawed in the biosafety cabinet. The virus was diluted with assay medium such that the amount of virus added to each well in a volume of 100 μ Ι _ was the amount judged to give a 85% to 95% cell kill rate 4 days after infection. Cell control containing medium only, virus infection control containing medium and virus, cytotoxicity control containing medium, each at 37 deg.C and 5% CO 2Four days after incubation, 4 hours at 37 ℃ followed by passage through the platesDye XTT reduction was measured and inhibition of CPE (increased cell viability) was measured spectrophotometrically at 450nm (reference wavelength 650nm) using Softmax Pro 4.6 software. The percent CPE reduction for virus-infected wells and percent cell viability for uninfected drug-control wells were calculated by four-parameter curve fitting analysis using microsoft Excel XLfit 4.
(2) PB2 binding assays homogeneous, high throughput binding/displacement assays were designed for the determination of IC50 for PB2 inhibitors. The principle of the assay reverts to the competitive displacement of fluorescent probes (m7GTP analogs) from their complexes with PB2 by PB2 inhibitors. Briefly, purified PB2 was incubated with fluorescent m7GTP analog and test inhibitors were added to the reaction mixture. The displacement of fluorescent m7GTP was read using a Perkin Elmer Envision. The properties are reported in table B below: +++++: IC50, <5 μ M; ++++: IC50, 5-25. mu.M; +++: IC50, 25-50. mu.M; ++: IC50, 50-100. mu.M; and +: IC50, > 100. mu.M
TABLE B
PB2 binding assays of several influenza a strains were also compared. IC50 values are reported in table C below: +++++: EC50, <5 μ M; ++++: EC50, 5-25 μ M; +++: EC50, 25-50 μ M; ++: EC50, 50-100. mu.M; and +: EC50, >100 μ M, -: not tested.
Watch C