COMPOUND FOR TARGETING AND DEGRADING PROTEIN, AND PREPARATION METHOD THEREFOR AND USE THEREOF

Abstract

A class of bifunctional compounds for targeting and degrading an IRAK4 kinase protein, a pharmaceutical composition and a preparation method therefor are provided. The compounds not only effectively inhibits and/or degrades the IRAK4 kinase protein in cells, but also effectively inhibit the production of IL-6 by immune cells, and has good degradation selectivity. The compounds can be used in the preparation of a drug for treating and/or preventing IRAK4-mediated related diseases or conditions, such as cancer, immunological diseases and inflammatory diseases.

Claims

1-33. (canceled)

34. A compound of formula I, and/or a stereoisomer, an enantiomer, a diastereomer, a deuterate, a hydrate, a solvate, a prodrug and/or a pharmaceutically acceptable salt thereof:
PTM-L-ULM   I wherein: PTM is a small molecule compound that can inhibit IRAK4 kinase protein or bind to IRAK4 kinase protein; L is a connecting chain, which connects PTM and ULM through a covalent bond; ULM is a small molecule ligand in E3 ubiquitin ligase complex, and the ULM has the following structure: ##STR00670## wherein, in ULM-1: X″ is CH or N; Y″ is CH, N, O or S; Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4 and Q.sub.5 are each independently CR.sub.3″ or N; R.sub.3″ are each independently hydrogen, deuterium, hydroxyl, amino, cyano, halogen, C1-C6 alkyl, C3-C8 cycloalkyl, 3-8-membered heterocycloalkyl, 6-10-membered aryl, 5-10-membered heteroaryl, —O(C1-C6 alkyl), —O—(C3-C8 cycloalkyl), —O-(3-8-membered heterocycloalkyl), —N(C1-C6 alkyl).sub.2, —NH(C3-C8 cycloalkyl)), —NH(3-8-membered heterocycloalkyl), —O-(6-10-membered aryl), or —O-(5-10-membered heteroaryl); and the alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted by 1-3 groups independently selected from hydroxyl, halogen, cyano, or amino; or R.sub.3″ together with its attached atoms form a cycloalkyl, heterocycloalkyl, aryl or heteroaryl; m″ is 1, 2 or 3; R.sub.1″ are each independently hydrogen, deuterium, hydroxyl, amino, cyano, halogen, C1-C6 alkyl, C3-C8 cycloalkyl, 3-8-membered heterocycloalkyl, 6-10-membered aryl, 5-10-membered heteroaryl, or —O(C1-C6 alkyl); and the alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted by 1-3 groups independently selected from hydroxyl, halogen, cyano, or amino; R.sub.2″ is absent, hydrogen, deuterium, C1-C6 alkyl, or C3-C6 cycloalkyl, the C1-C6 alkyl and C3-C6 cycloalkyl are optionally substituted by 1-3 groups independently selected from hydroxyl, halogen, —O—(C═O)—(C1-C6 alkyl), cyano or amino.

35. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein one or two of Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4, and Q.sub.5 in ULM-1 is N, the rest are each independently CR.sub.3″.

36. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4, and Q.sub.5 in ULM-1 are each independently CR.sub.3″.

37. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein Y″ in ULM-1 is N.

38. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.1″ in ULM-1 are each independently hydrogen, deuterium, —F, —Cl, or C1-C6 alkyl, the alkyl is optionally substituted by 1-3 halogens; preferably R.sub.1″ is hydrogen.

39. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.2″ in ULM-1 is hydrogen or C1-C6 alkyl, and the alkyl is optionally substituted by 1-3 halogens; preferably R.sub.2″ is hydrogen.

40. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.3″ in ULM-1 is each independently hydrogen, deuterium, halogen, —O(C1-C6 alkyl), or C1-C6 alkyl, and the alkyl is optionally substituted by 1 to 3 halogens; preferably R.sub.3″ is each independently hydrogen, deuterium, F, Cl, methyl, methoxy, ethoxy, trifluoromethoxy, 2-hydroxypropyl-2-yl or trifluoromethyl.

41. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein ULM is selected from the group consisting of: ##STR00671## wherein, Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4, Q.sub.5, R.sub.1″, R.sub.2″, R.sub.3″ and m″ are as defined in claim 34.

42. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein ULM is selected from the group consisting of: ##STR00672## ##STR00673## wherein, Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4, Q.sub.5, R.sub.1″, R.sub.2″, R.sub.3″ and m″ are as defined in claim 34.

43. The compound of formula I according to claim 41, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein ULM is selected from ##STR00674## ##STR00675##

44. The compound of formula I according to claim 42, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein ULM is selected from ##STR00676## ##STR00677##

45. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein PTM has the following structure: ##STR00678## wherein, in PTM-68: Z.sub.1 is absent, substituted or unsubstituted C1-C6 alkylene, substituted or unsubstituted C3-C6 cycloalkylene, substituted or unsubstituted C2-C6 alkenylene, or substituted or unsubstituted C2-C6 alkynylene; Z.sub.2 is carbonyl, ##STR00679## R.sub.1 is ##STR00680## each R.sub.2 and each R.sub.3 are independently absent, substituted or unsubstituted C1-C10 alkyl, substituted C3-C10 cycloalkyl, halogen, substituted or unsubstituted C1-C10 haloalkyl, substituted or unsubstituted C1-C12 hydroxyalkyl, substituted or unsubstituted C1-C12 sulfydrylalkyl, substituted or unsubstituted C3-C12 hydroxycycloalkyl, substituted or unsubstituted C3-C12 sulfydrylcycloalkyl, cyano, nitro, substituted or unsubstituted 3-12-membered heterocycloalkyl, -A-R.sub.10, or —N(R.sub.11)R.sub.12; R.sub.4 is substituted or unsubstituted C6-C20 aryl, or substituted or unsubstituted 5-20-membered heteroaryl; R.sub.5 is substituted or unsubstituted C1-C10 alkyl, or substituted or unsubstituted C3-C10 cycloalkyl; R.sub.6 is substituted or unsubstituted C1-C10 alkyl, or substituted or unsubstituted C3-C10 cycloalkyl; R.sub.7 is hydrogen, deuterium, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-C10 cycloalkyl, or R.sub.13—C(O)—; R.sub.8 and R.sub.9 are each independently substituted or unsubstituted C1-C10 alkyl, or R.sub.8 and R.sub.9 together with S atom to which they are attached form a 3-12 membered heterocycloalkyl; R.sub.10 is hydrogen, deuterium, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-12-membered heterocycloalkyl, or —R.sub.14-R.sub.15; R.sub.11 and R.sub.12 are each independently hydrogen, deuterium, substituted or unsubstituted C1-C10 alkyl, or substituted or unsubstituted C3-C8 cycloalkyl; R.sub.13 is substituted or unsubstituted C1-C10 alkyl, or substituted or unsubstituted C3-C10 cycloalkyl; R.sub.14 is substituted or unsubstituted C1-C6 alkylene; R.sub.15 is substituted or unsubstituted C3-C12 cycloalkyl, or substituted or unsubstituted C3-C12 heterocycloalkyl; wherein, any “substituted” refers to one or more (preferably 1, 2, 3 or 4) hydrogen atoms on the group is substituted by a substituent selected from the group consisting of: C2-C8 acyl, C3-C8 cycloalkyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, hydroxyl, sulfydryl, amino, nitro, halogen, 3-12 membered heterocycloalkyl, cyano, C1-C10 haloalkyl, C3-C8 halocycloalkyl, C2-C4 ester, C2-C4 amide, C1-C4 carboxyl, C2-C6 alkenyl, C2-C6 alkynyl, C6-C12 aryl, 5-12 membered heteroaryl, —N(R.sub.16)R.sub.17; R.sub.16 and R.sub.17 are each independently hydrogen, deuterium, C1-C6 alkyl, C3-C8 cycloalkyl, or 3-membered heterocycloalkyl; the heterocycloalkyl, heteroaryl and heteroalkyl ring are each independently have 1-3 (preferably 1, 2 or 3) heteroatoms selected from N, O and S; A is S or O; a is 0 or 1, b is 0, 1, 2 or 3.

46. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein PTM has the following structure: ##STR00681## or a pharmaceutically acceptable salt or a stereoisomer thereof, wherein in the PTM-4, PTM-4′, PTM-4c, and PTM-4c′, A is optionally substituted heteroaryl, optionally substituted aryl, optionally substituted heterocycloalkyl, optionally substituted cycloalkyl, optionally substituted (cycloalkyl) alkyl, optionally substituted (heterocycloalkyl) alkyl, optionally substituted (aryl) alkyl-, optionally substituted (heteroaryl) alkyl-, optionally substituted cycloalkyl-NR.sup.X—, optionally substituted heterocycloalkyl-NR.sup.X—, optionally substituted aryl-NR.sup.X—, optionally substituted heteroaryl-NR.sup.X—, optionally substituted cycloalkyl-O—, optionally substituted heterocycloalkyl-O—, optionally substituted aryl-O—, or optionally substituted heteroaryl-O—; wherein the optional substituent is R.sup.X; B is hydrogen, deuterium, halogen, cyano, optionally substituted alkyl, alkenyl, optionally substituted alkoxy, —NR.sup.aR.sup.b, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocycloalkyl, optionally substituted heteroaryl, optionally substituted (cycloalkyl) alkyl, optionally substituted (heterocycloalkyl) alkyl, optionally substituted (aryl) alkyl-, optionally substituted (heteroaryl) alkyl-, optionally substituted cycloalkyl-NR.sup.x—, optionally substituted heterocycloalkyl-NR.sup.x—, optionally substituted aryl-NR.sup.x—, optionally substituted heteroaryl-NR.sup.x—, optionally substituted cycloalkyl-O—, optionally substituted heterocycloalkyl-O—, optionally substituted heteroaryl-O—, optionally substituted aryl-O—; wherein the optional substituent is R.sup.y; Q is absent or optionally substituted heterocycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted (heterocycloalkyl) alkyl, optionally substituted (heteroaryl) alkyl, optionally substituted (aryl) alkyl-, optionally substituted (cycloalkyl) alkyl, —NR.sup.3R.sup.4, —OR.sup.3 or —SR.sup.3; wherein the optional substituent is R.sup.z; W is N or CH; R.sub.1 is hydrogen, deuterium, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted (cycloalkyl) alkyl, optionally substituted (heterocycloalkyl) alkyl, optionally substituted heterocycloalkyl, optionally substituted (aryl) alkyl-, optionally substituted (heteroaryl) alkyl-, optionally substituted alkoxyalkyl, optionally substituted aminoalkyl or —(CH.sub.2).sub.mR.sub.2; wherein the optional substituent is each independently selected from halogen, hydroxyl, alkoxy, amino, nitro, cycloalkyl, aryl, heteroaryl or heterocycloalkyl; R.sub.2 is hydrogen, deuterium, —NR.sup.aR.sup.b, alkoxy, hydroxyl, optionally substituted heteroaryl or optionally substituted heterocycloalkyl; wherein the optional substituent is R.sup.y; R.sub.3 and R.sub.4 are each independently selected from optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocycloalkyl, optionally substituted (aryl) alkyl-, optionally substituted (cycloalkyl) alkyl, optionally substituted (heteroaryl) alkyl, or optionally substituted (heterocycloalkyl) alkyl; wherein the optional substituent is each independently selected from alkyl, halogen, haloalkyl, hydroxyl, hydroxyalkyl, alkoxy, alkoxy alkyl, amino, nitro, cycloalkyl, (cycloalkyl) alkyl, aryl, (aryl) alkyl-, (heteroaryl) alkyl-, (heterocycloalkyl) alkyl, heteroaryl and (heteroaryl) alkyl; each R.sup.a and R.sup.b are independently selected from hydrogen, deuterium, alkyl, aminoalkyl, acyl, or heterocycloalkyl; or R and R.sup.b together with the nitrogen to which they are attached form an optionally substituted ring; R.sup.X is hydrogen, deuterium, alkyl, hydroxyl, hydroxyalkyl, acyl or cycloalkyl; each R.sup.y and R.sup.z are independently selected from hydroxyl, hydroxyalkyl, halogen, alkyl, oxo, haloalkyl, alkoxy, alkenyloxy, amino, nitro, cyano, —SH, —S(alkyl), glycine ester, ester, thioester, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, (cycloalkyl) alkyl, (heterocycloalkyl) alkyl, (aryl) alkyl- and (heteroaryl) alkyl; wherein the hydroxyl, hydroxyalkyl, alkoxy, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally further substituted by one or more substituents selected from alkyl, halogen, alkenyl, amino, nitro, cycloalkyl or (cycloalkyl) alkyl; or R.sup.y and R.sup.z together with the atoms to which they are attached form an alkyl chain with 1-10 carbon atoms; 1-3 carbon atoms of which are optionally substituted by O, NH or S; m is 1, 2 or 3; and n is 1 or 2.

47. The compound of formula I according to claim 45, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein PTM is ##STR00682## wherein, in PTM-71: ring A is 6-10 membered aryl or 5-10 membered heteroaryl; R.sub.d is each independently hydrogen, deuterium, halogen, cyano, C1-C6 alkyl, C3-C6 cycloalkyl, or 5-10-membered heteroaryl; and the alkyl, cycloalkyl, and heteroaryl are optionally substituted by one or more groups selected from halogen, hydroxyl, or amino; n is 1, 2, 3 or 4; R.sub.e is hydrogen or C1-C6 alkyl; R.sub.e is hydrogen, deuterium, —O—(C1-C6 alkyl), —O—(C3-C8 cycloalkyl), —O-(3-8-membered heterocycloalkyl), —O-(6-10-membered aryl), —O-(5-10-membered heteroaryl), —N(C1-C6 alkyl).sub.2, —NH(C3-C8 cycloalkyl), —NH(3-8-membered heterocycloalkyl), —NH-(6-10 membered aryl), —NH-(5-membered heteroaryl), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3-8-membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl; and the alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl are optionally substituted by one or more groups independently selected from hydroxyl, amino, halogen, cyano or —O—(C1-C6 alkyl); R.sub.b is hydrogen, deuterium, —O—(C1-C6 alkyl), —O—(C3-C8 cycloalkyl), —O-(3-8-membered heterocycloalkyl), —O-(6-10-membered aryl), —O-(5-10-membered heteroaryl), —N(C1-C6 alkyl).sub.2, —NH(C3-C8 cycloalkyl), —NH(3-8-membered heterocycloalkyl), —NH-(6-10 membered aryl), —NH-(5-membered heteroaryl), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3-8-membered heterocycloalkyl, 6-10 membered aryl or 5-10 membered heteroaryl; and the alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted by one or more groups independently selected from hydroxyl, amino, halogen, or cyano; R.sub.a is hydrogen, deuterium, 3-8-membered heterocycloalkyl, C3-C8 cycloalkyl, 6-10 membered aryl, 5-10 membered heteroaryl, C1-C6 alkyl, ##STR00683## the alkyl is optionally substituted by one or more substituents selected from halogen, hydroxyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, S(O).sub.1-2(C1-C6 alkyl), S(O).sub.1-2(C3-C6 cycloalkyl), unsubstituted or mono- or polyhalogen-substituted C3-C6 cycloalkyl, unsubstituted or mono- or di-methyl substituted monocyclic saturated heterocycloalkyl having 4-6 ring atoms and containing heteroatoms or heterogroups selected from O, S, SO or SO.sub.2; and the cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are optionally substituted by one or more groups selected from hydroxyl, halogen, amino, cyano, C1-C6 alkyl, —O(C1-C6 alkyl), halogenated C1-C6 alkyl, hydroxyl C1-C6 alkyl, or amino C1-C6 alkyl.

48. The compound of formula I according to claim 47, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein PTM is ##STR00684## wherein, in PTM-71: ring A is phenyl or pyridyl; R.sub.d is each independently hydrogen, deuterium, halogen, cyano, C1-C6 alkyl, or C3-C6 cycloalkyl; and the alkyl and cycloalkyl are optionally substituted by one or more groups selected from halogen, hydroxyl or amino; n is 1 or 2; R.sub.e is hydrogen; R.sub.e is hydrogen, deuterium, —O(C1-C6 alkyl), —N(C1-C6 alkyl).sub.2, C1-C6 alkyl, —O(C3-C6 cycloalkyl), —NH(C3-C6 cycloalkyl), —O(3-6-membered heterocycloalkyl), —NH (3-6-membered heterocycloalkyl); and the alkyl, cycloalkyl and heterocycloalkyl are optionally substituted by one or more groups independently selected from hydroxyl, amino, halogen, cyano or —O—(C1-C6 alkyl); R.sub.b is hydrogen or C1-C6 alkyl, and the alkyl is optionally substituted by one or more groups independently selected from hydroxyl, amino, halogen or cyano; R.sub.a is hydrogen, deuterium, 3-8-membered heterocycloalkyl, C3-C8 cycloalkyl, C1-C6 alkyl, ##STR00685## the alkyl is optionally substituted by one or more substituents selected from halogen, hydroxyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, S(O).sub.1-2(C1-C6 alkyl), S(O).sub.1-2(C3-C6 cycloalkyl), unsubstituted or mono- or polyhalogen-substituted C3-C6 cycloalkyl, unsubstituted or mono- or di-methyl substituted monocyclic saturated heterocycloalkyl having 4-6 ring atoms and containing heteroatoms or heterogroups selected from O, S, SO or SO.sub.2; the cycloalkyl and the heterocycloalkyl are optionally substituted by one or more groups selected from hydroxyl, halogen, amino, cyano, C1-C6 alkyl, —O(C1-C6 alkyl), halogenated C1-C6 alkyl, hydroxyl C1-C6 alkyl, or amino C1-C6 alkyl.

49. The compound of formula I according to claim 48, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.d in PTM-71 is each independently hydrogen, deuterium, halogen or C1-C6 alkyl; and the alkyl is optionally substituted by one or more groups selected from halogen or hydroxyl; preferably R.sub.d is hydrogen, deuterium, F, methyl, difluoromethyl, trifluoromethyl or 2-hydroxypropyl.

50. The compound of formula I according to claim 48, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.c in PTM-71 is C1-C6 alkyl, and the alkyl is optionally substituted by one or more groups independently selected from hydroxyl and halogen; preferably R.sub.c is difluoromethyl or 2-hydroxypropyl.

51. The compound of formula I according to claim 50, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.c in PTM-71 is C1-C6 alkyl, and the alkyl is substituted by hydroxyl.

52. The compound of formula I according to claim 48, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.c in PTM-71 is —O(C1-C6 alkyl), and the alkyl is optionally substituted by one or more groups independently selected from hydroxyl, halogen or —OCH.sub.3; preferably R.sub.c is methoxy, ethoxy, isopropoxy, —OCH.sub.2CH.sub.2OCH.sub.3, difluoromethoxy or trifluoromethoxy.

53. The compound of formula I according to claim 48, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.c in PTM-71 is —O-(3-6-membered heterocycloalkyl containing one or two heteroatoms selected from N or O); preferably ##STR00686##

54. The compound of formula I according to claim 48, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.b in PTM-71 is hydrogen or methyl.

55. The compound of formula I according to claim 48, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.a in PTM-71 is 3-12 membered heterocycloalkyl containing one or two heteroatoms selected from N, O or S, and the heterocycloalkyl is optionally substituted by one or more groups selected from hydroxyl, halogen, amino, cyano, C1-C6 alkyl, —O(C1-C6 alkyl), halogenated C1-C6 alkyl, hydroxyl C1-C6 alkyl, or amino C1-C6 alkyl; preferably R.sub.a is 3-12-membered heterocycloalkyl containing one or two heteroatoms selected from N or O, and the heterocycloalkyl is optionally substituted by one or more groups selected from hydroxyl, halogen, amino, cyano, C1-C6 alkyl, —O(C1-C6 alkyl), halogenated C1-C6 alkyl, hydroxyl C1-C6 alkyl, or amino C1-C6 alkyl; more preferably R.sub.a is piperidinyl or piperazinyl, the piperidinyl and the piperazinyl are optionally substituted by one or more groups selected from hydroxyl, halogen, amino, cyano, C1-C6 alkyl, —O(C1-C6 alkyl), halogenated C1-C6 alkyl, hydroxyl C1-C6 alkyl, or amino C1-C6 alkyl; most preferably is piperidinyl which is optionally substituted by one or more groups selected from hydroxyl, amino, methyl, methoxy, hydroxymethyl, or trifluoromethyl.

56. The compound of formula I according to claim 48, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein R.sub.a in PTM-71 is cyclohexane, and the cyclohexane is optionally substituted by one or more groups selected from hydroxyl, halogen, amino, cyano, C1-C6 alkyl, —O(C1-C6 alkyl), halogenated C1-C6 alkyl, hydroxyl C1-C6 alkyl, or amino C1-C6 alkyl, preferably optionally substituted by one or more groups selected from hydroxyl, amino, methyl, methoxy, hydroxymethyl, or trifluoromethyl.

57. The compound of formula I according to claim 48, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein PTM is ##STR00687## ##STR00688## ##STR00689##

58. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein L is a bond.

59. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein L is —(CH.sub.2).sub.j—, and one or more methylenes in —(CH.sub.2).sub.j— are optionally replaced by a group selected from —NR.sup.3′—, —O—, —S—, —S(O)—, —S(O)NR.sup.3′—, —NR.sup.3′S(O)—, —S(O).sub.2—, —S(O).sub.2NR.sup.3′—, —NR.sup.3′S(O).sub.2—, —NR.sup.4′S(O).sub.2NR.sup.3′—, —CR.sup.1′R.sup.2′—, —C(O)—, —C(O)O—, —OC(O)—, —NR.sup.3′C(O)O—, —OC(O)NR.sup.3′—, —C(O)NR.sup.3′—, —NR.sup.3′C(O)—, —NR.sup.4′C(O)NR.sup.3′—, —P(O)—, —P(O)O—, —OP(O)—, —OP(O)O—, vinylidene, ethynylene, C3-C12 cycloalkylene, 3-12 membered heterocycloalkylene containing one or more heteroatoms selected from N, O or S, 6-10 membered arylene or 5-10 membered heteroarylene; and the vinylidene, cycloalkylene, heterocycloalkylene, arylene, and heteroarylene are each independently optionally substituted by one or more substituents selected from halogen, —OR.sup.3′, —NR.sup.3′R.sup.4′, oxo, nitro, cyano, C1-C6 alkyl, —S(C1-C6 alkyl), C3-C10 cycloalkyl, 3-10 membered heterocycloalkyl, 6-10 membered aryl, 5-10 membered heteroaryl, —C(O)R.sup.1′, —C(O)OR.sup.3′, —OC(O)R.sup.1′, —C(O)NR.sup.3′, —NR.sup.3′C(O)R.sup.1′, —S(O)R.sup.1′, —S(O)NR.sup.3′, —S(O)).sub.2R.sup.1′, —S(O).sub.2NR.sup.3′, —NR.sup.3′S(O).sub.2R.sup.1′, —NR.sup.4′S(O).sub.2NR.sup.3′, —OC(O)NR.sup.3′, —NR.sup.4′C(O)NR.sup.3′, and the alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are each independently optionally substituted by one or more substituents selected from halogen, —OH, —NR.sup.3R.sup.4′, oxo, nitro, cyano, C1-C6 alkyl, C3-C10 cycloalkyl, 3-10 membered heterocycloalkyl, 6-10 membered aryl, or 5-10 membered heteroaryl; R.sup.1′ and R.sup.2′ are each independently halogen, —OH, —NR.sup.3′R.sup.4′, C1-C6 alkyl, chlorinated C1-C6 alkyl, hydroxyl C1-C6 alkyl, —O(C1-C6 alkyl), —NH(C1-C6 alkyl), —NH(C1-C6 alkyl), C3-C10 cycloalkyl, —O(C3-C10 cycloalkyl), —NH(C3-C10 cycloalkyl), 3-10 membered heterocycloalkyl, —O(3-10 membered heterocycloalkyl), —NH(3-10 membered heterocycloalkyl), 6-10 membered aryl, —O(6-10 membered aryl), —NH(6-10 membered aryl), 5-10 membered heteroaryl, —O(5-10 membered heteroaryl), or —NH(5-10 membered heteroaryl); R.sup.3′ and R.sup.4′ are each independently hydrogen, deuterium, C1-C6 alkyl, C3-C10 cycloalkyl, 3-10 membered heterocycloalkyl, 6-10 membered aryl, or 5-10 membered heteroaryl; j is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25.

60. The compound of formula I according to claim 59, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein L is —(CH.sub.2).sub.j, 1, 2, 3 or 4 methylenes in —(CH.sub.2).sub.j— are optionally replaced by a group selected from —NH—, —NCH3-, —NCH2CH.sub.3—, —O—, —C(CH.sub.3).sub.2—, —CHF—, —CHCF.sub.3—, —C(O)—, —C(O)O—, —OC(O)—, —C(O)NH—, —C(O)NCH.sub.3—, —NHC(O)—, —NCH.sub.3C(O)—, vinylidene, ethynylene, cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylidene, oxiranylene, oxetanylene, oxolanylene, oxanilene, azridinylene, azetidinylene, azacyclopentylene, piperidinylidene, piperazinylidene, morpholinylidene, perhomomorpholinylidene, phenylene, pyrrolylidene, thienylidene, furanylidene, imidazolylidene, pyrazolylidene, triazolylidene, tetrazolylidene, oxazolylidene, isoxazolylidene, thiazolylidene, isothiazolylidene, pyridylidene, pyrimidinylidene, pyridazinylidene, pyrazinylidene, ##STR00690## and the group is optionally substituted by one or more substituents selected from halogen, oxo, —NR.sup.3′R.sup.4′, —OR.sup.3′, or C1-C4 alkyl, the alkyl is optionally substituted by one or more substituents selected from halogen, —OH, or —NH.sub.2, R.sup.3′ and R.sup.4′ are each independently hydrogen, deuterium, C1-C4 alkyl, and j is 2, 3, 4, 5, 6, 7 or 8.

61. The compound of formula I according to claim 59, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein L is —(CH.sub.2).sub.j—, 1, 2 or 3 methylenes in —(CH.sub.2).sub.j— are optionally substituted by a group selected from —O—, —NH—, —NCH.sub.3—, —NCH.sub.2CH.sub.3—, —C(O)—, —C(O)NH—, —NHC(O)—, —NCH.sub.3C(O)—, —C(O)NCH.sub.3—, azridinylene, azetidinylene, azacyclopentylene, piperidinylidene, piperazinylidene, ##STR00691## is 2, 3, 4, 5, 6, 7, or 8.

62. The compound of formula I according to claim 59, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein L is —(CH.sub.2).sub.j-1—C(O)—, the methylene in —(CH.sub.2).sub.j-1—C(O)— is as defined in claim 59, optionally substituted by one or more groups, j is as defined in claim 59.

63. The compound of formula I according to claim 59, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein L is ##STR00692## ##STR00693## ##STR00694## ##STR00695## ##STR00696## ##STR00697## ##STR00698## ##STR00699## ##STR00700## ##STR00701## ##STR00702## ##STR00703## ##STR00704## ##STR00705## ##STR00706## ##STR00707## ##STR00708## ##STR00709## ##STR00710## ##STR00711## ##STR00712## ##STR00713## ##STR00714## ##STR00715## ##STR00716## ##STR00717## ##STR00718## ##STR00719## ##STR00720## ##STR00721## ##STR00722## ##STR00723## ##STR00724## ##STR00725## ##STR00726## ##STR00727## ##STR00728## ##STR00729## ##STR00730## ##STR00731## ##STR00732## ##STR00733## ##STR00734## ##STR00735## ##STR00736## ##STR00737##

64. The compound of formula I according to claim 59, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein L is LA, ##STR00738## wherein, in LA, ring D is absent or is C3-C12 cycloalkylene or 3-12 membered heterocycloalkylene containing 1-2 heteroatoms selected from N, O or S, the cycloalkylene and the heterocycloalkylene are optionally substituted by a substituent selected from halogen, oxo, cyano, amino, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl or —O—(C1-C6 alkyl); ring B is absent or is C3-C12 cycloalkylene or 3-12 membered heterocycloalkylene containing 1-2 heteroatoms selected from N, O or S, the cycloalkylene and the heterocycloalkylene are optionally substituted by a substituent selected from halogen, oxo, cyano, amino, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl or —O—(C1-C6 alkyl); ring C is absent, C3-C12 cycloalkylene or 3-12 membered heterocycloalkylene containing 1-2 heteroatoms selected from N, O or S, the cycloalkylene and the heterocycloalkylene are optionally substituted by a substituent selected from halogen, oxo, cyano, amino, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl or —O—(C1-C6 alkyl); X″″ is a bond, —NH—, —NCH.sub.3—, —O—, —C(CH.sub.3).sub.2—, —S—, —C═C—, —C≡C—, —CHF—, —CHCF.sub.3—, —C(O)—, —S(O)—, —S(O).sub.2—, —C(O)O—, —OC(O)—, —C(O)NH—, —C(O)NCH.sub.3—, —NHC(O)— or —NCH.sub.3C(O)—; L.sub.3 is —(CH.sub.2).sub.k, one or two methylenes in L3 are optionally substituted by a substituent selected from —O—, —NH—, —N(C1-C6 alkyl)-, —N(C1-C6 haloalkyl)-, —N(hydroxyC1-C6 alkyl)- or —N(C3-C8 cycloalkyl)-, k is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

65. The compound of formula I according to claim 59, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein L is LA, ##STR00739## wherein, in LA, ring D and ring B are absent, ring C is 4-7-membered saturated monocyclic heterocycloalkylene containing 1 or 2 nitrogen heteroatoms, or 7-11-membered spiroheterocycloalkylene containing 1 or 2 nitrogen heteroatoms; X″″ is —C(O)—; L.sub.3 is —(CH.sub.2).sub.k, one methylene in L.sub.3 is optionally replaced by a group selected from —O—, —NH—, —NCH.sub.3— or —NCH.sub.2CH.sub.3—; and k is 2, 3 or 4.

66. The compound of formula I according to claim 59, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein L is ##STR00740## ##STR00741## ##STR00742## ##STR00743##

67. The compound of formula I according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, wherein the compound of formula I is ##STR00744## ##STR00745## ##STR00746## ##STR00747## ##STR00748## ##STR00749## ##STR00750## ##STR00751## ##STR00752## ##STR00753## ##STR00754## ##STR00755## ##STR00756## ##STR00757## ##STR00758## ##STR00759## ##STR00760## ##STR00761## ##STR00762## ##STR00763## ##STR00764## ##STR00765## ##STR00766## ##STR00767## ##STR00768## ##STR00769## ##STR00770## ##STR00771## ##STR00772## ##STR00773## ##STR00774## ##STR00775## ##STR00776## ##STR00777## ##STR00778## ##STR00779## ##STR00780## ##STR00781## ##STR00782## ##STR00783## ##STR00784##

68. A pharmaceutical composition comprising the compound according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

69. A method for the treatment and/or prevention of IRAK4-mediated disease or condition, or TLR (other than TLR3R), IL-1α or IL-1β receptor family mediated disease or condition comprising the step of administrating the compound according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof into a subject in need thereof.

70. A method for the treatment and/or prevention of cancer, neurodegenerative diseases, viral diseases, autoimmune diseases, inflammatory diseases, hereditary diseases, hormone-related diseases, metabolic diseases, organ transplantation-related diseases, immunodeficiency diseases, destructive bone diseases, proliferative disorders, infectious diseases, conditions related to cell death, thrombin-induced platelet aggregation, liver diseases, pathological immune conditions involving T cell activation, cardiovascular diseases or CNS diseases comprising the step of administrating the compound according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof into a subject in need thereof.

71. A method for the treatment and/or prevention of brain cancer, kidney cancer, liver cancer, adrenal cancer, bladder cancer, breast cancer, gastric cancer, ovarian cancer, colon cancer, rectal cancer, prostate cancer, pancreatic cancer, lung cancer, vaginal cancer, cervical cancer, testicular cancer, genitourinary cancer, esophageal cancer, laryngeal cancer, skin cancer, bone cancer, thyroid cancer, sarcoma, glioblastoma, neuroblastoma, multiple myeloma, gastrointestinal cancer, neck or head tumor, epidermal hyperhyperplasia, bovine skin moss, prostate hyperplasia, Adenoma, adenocarcinoma, keratoacanthoma, epidermoid cancer, large cell carcinoma, non-small cell lung cancer, lymphoma, Hodgkin's and non-Hodgkin's, breast cancer, follicular cancer, undifferentiated tumor, papillary tumor, seminoma, melanoma, ABC DLBCL, Hodgkin's lymphoma, primary cutaneous T-cell lymphoma, chronic lymphocytic leukemia, smoking indolent multiple myeloma, leukemia, diffuse large B-cell lymphoma DLBCL, chronic lymphocytic leukemia CLL, chronic lymphocytic lymphoma, primary exudative lymphoma, Burkitt lymphoma/leukemia, acute lymphocytic leukemia, pre-B cell lymphocytic leukemia, lymphoplasmic lymphoma, Waldenstroms's macroglobulinemia WM, splenic marginal zone lymphoma, multiple myeloma, or plasmacytoma or intravascular large B-cell lymphoma, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, cerebral ischemia or traumatic injury, glutamate neurotoxicity, hypoxia, epilepsy, diabetes treatment, metabolic syndrome, obesity, neurodegenerative diseases caused by organ transplantation or graft-versus-host disease, eye disease, such as eye allergy, conjunctivitis, dry eye or spring conjunctivitis, diseases affecting the nose, including allergic rhinitis; autoimmune hematological diseases, such as hemolytic anemia, aplastic anemia, pure red blood cell anemia and idiopathic thrombocytopenia, systemic lupus erythematosus, rheumatoid arthritis, polychondritis, scleroderma, dermatomyositis, polymyositis, chronic active hepatitis, myasthenia gravis, Stephen-Johnson syndrome, idiopathic stomatitis diarrhea, autoimmune inflammatory bowel disease, bowel syndrome, celiac disease, root periostitis, lung hyaline membrane disease, nephropathy, glomerular disease, Alcoholic liver disease, multiple sclerosis, endocrine ophthalmopathy, Grave's disease, Sarcomatosis, dry eye, spring conjunctival keratitis, interstitial pulmonary fibrosis, bovine moss arthritis, systemic juvenile idiopathic arthritis, nephritis, vasculitis, interstitial cystitis, diverticulitis, Glomerulonephritis, chronic granulomatous disease, endometriosis, leptospirosis nephropathy, glaucoma, retinal disease, aging, headache, pain, complex regional pain syndrome, cardiac hypertrophy, muscle atrophy, catabolism, obesity, slow fetal growth, hypercholesterolemia, heart disease, chronic heart failure, mesothelioma, anhidromic ectodermal dysplasia, Behcet's disease, pigment incontinence, Paget's disease, pancreatitis, hereditary periodic fever syndrome, asthma, acute lung injury, acute respiratory distress syndrome, eosinophilia, allergic reaction, systemic allergic reaction, sinusitis, eye allergy, silica-induced diseases, COPD, lung disease, cystic fibrosis, acid-induced lung injury, pulmonary hypertension, polyneuropathy, Cataract, muscle inflammation combined with systemic sclerosis, inclusion body myositis, myasthenia gravis, thyroiditis, Addison's disease, lichen planus, type 1 diabetes, type 2 diabetes, appendicitis, atopic dermatitis, asthma, allergies, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, chronic transplant rejection, colitis, conjunctivitis, cystitis, lacrimal gland inflammation, dermatitis, dermatomyositis, polymyositis, encephalitis, endocarditis, endometritis, enteritis, enterocolitis, upper ankle inflammation, epididymitis, fasciitis, fibrous tissue inflammation, gastritis, gastroenteritis, Henoch-Schonlein purpura, hepatitis, suppurative sweat Inflammation, immunoglobulin A nephropathy, interstitial lung disease, laryngitis, mastitis, meningitis, myelitis, myocarditis, myositis, nephritis, liver fibrosis, renal fibrosis, alcoholic fatty liver, non-alcoholic fatty liver, heart fibrosis, psoriasis, Crohn's disease, inflammatory bowel disease, oophoritis, orchitis, osteitis, otitis, pancreatitis, mumps, pericarditis, peritonitis, pharyngitis, pleurisy, phlebitis, local pneumonia, pneumonia, polymyositis, proctitis, prostatitis, pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, articular inflammation, tendinitis, tonsillitis, ulcerative colitis, uveitis, vaginitis, vasculitis, vulvitis, alopecia areata, erythema multiforme, dermatitis herpetiformis, sclerosis, vitiligo, hypersensitivity vasculitis, urticaria, bullous pemphigoid, pemphigus vulgaris, deciduous pemphigus, paraneoplastic pemphigus, acquired bullous epidermal laxity, acute and chronic gout, chronic gouty arthritis, bovine skin moss, bovine skin arthritis, rheumatoid arthritis, juvenile rheumatoid arthritis, cryopyrin-associated periodic syndrome or osteoarthritis diseases comprising the step of administrating the compound according to claim 34, and/or the stereoisomer, the enantiomer, the diastereomer, the deuterate, the hydrate, the solvate, the prodrug and/or the pharmaceutically acceptable salt thereof into a subject in need thereof.

Description

DETAILED DESCRIPTION

[1310] Unless stated to the contrary, the following terms used in the specification and claims have the following meanings.

[1311] “Alkyl” refers to saturated aliphatic hydrocarbon groups, including linear or branched alkyls; C1-C8 alkyl refers to alkyls containing 1-8 carbon atoms, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-amyl, 1, 1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 1, 2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl or its various branched isomers; preferably C1-C6 alkyl; more preferably C1-C4 alkyl. The alkyl may be substituted or unsubstituted.

[1312] “Cycloalkyl” means saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituents; “C3-C12 cycloalkyl” means cycloalkyl comprising 3 to 12 carbon atoms; “C3-C8 cycloalkyl” means cycloalkyl comprising 3 to 8 carbon atoms; “C5-C10 cycloalkyl” means cycloalkyl comprising 5 to 10 carbon atoms;

[1313] Non-limiting examples of monocycloalkyl comprise cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclohepttrienyl, cyclooctyl, etc., preferably cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; preferably C3-C8 membered cycloalkyl; more preferably C3-C6 cycloalkyl.

[1314] Polycyclic cycloalkyls include spiro, fused and bridged cycloalkyls. “Spiro cycloalkyl” refers to a polycyclic group that shares a carbon atom (called a spiro atom) between monocyclyls. They can contain one or more double bonds, but none of the rings have a fully conjugated π electronic system. According to the number of spiro atoms shared between the rings, the spiro alkyl is divided into single spiro cycloalkyl, double spiro cycloalkyl or polyspiro cycloalkyl, preferably double spiro cycloalkyl. Non-limiting examples of spiro cycloalkyl include:

##STR00395##

[1315] “Fused cycloalkyl” refers to a fully carbon polycyclic group in which each ring in the system shares an adjacent pair of carbon atoms with the other rings in the system, wherein one or more rings may contain one or more double bonds, but none of them has a fully conjugated π-electron system. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl, preferably bicyclic fused cycloalkyl. Non-limiting examples of fused cycloalkyl include:

##STR00396##

[1316] “Bridged cycloalkyl” refers to an all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, they can contain one or more double bonds, but no ring has a fully conjugated π electron system. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl. Non-limiting examples of bridged cycloalkyl include:

##STR00397##

[1317] The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl, and non-limiting examples include indanyl, tetrahydronaphthyl, benzocycloheptyl, etc. The cycloalkyl may be optionally substituted or unsubstituted.

[1318] “Heterocycloalkyl” refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O)r (wherein r is an integer of 0, 1 or 2), but do not contain the ring moieties of —O—O—, —O—S— or —S—S—, and the remaining ring atoms are carbon. “3-12-membered heterocycloalkyl” refers to a cyclyl containing 3 to 12 ring atoms, “5-10-membered heterocycloalkyl” refers to a cyclyl containing 5 to 10 ring atoms, and “3-8-membered heterocycloalkyl” refers to a cyclyl containing 3 to 8 ring atoms, preferably “3-12-membered heterocycloalkyl” containing 1-3 heteroatoms selected from N, O or S, more preferably a 3-12-membered heterocycloalkyl containing 1 or 2 N atoms.

[1319] Non-limiting examples of monocyclic heterocycloalkyl include pyrrolidyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc., preferably piperidinyl and piperazinyl.

[1320] Polycyclic heterocycloalkyls include spiro, fused and bridged heterocycloalkyls. “Spiro heterocycloalkyl” refers to a polycyclic heterocycloalkyl that shares one atom (called a spiro atom) between single rings, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O)r (wherein r is an integer of 0, 1, 2), and the remaining ring atoms are carbon. They can contain one or more double bonds, but none of the rings have a fully conjugated π-electron system. According to the number of spiro atoms shared between the rings, the spiro cycloalkyl is divided into mono-spiro heterocycloalkyl, bis-spiro heterocycloalkyl or polyspiro heterocycloalkyl, preferably saturated “3-12 membered bis-spiro heterocycloalkyl” containing 1-3 heteroatoms selected from N, O or S; more preferably saturated “3-12 membered bis-spiro heterocycloalkyl” containing 1 or 2 N atoms. Non-limiting examples of spiro heterocycloalkyl include:

##STR00398##

[1321] “Fused heterocycloalkyl” refers to a polycyclic heterocycloalkyl in which each ring in the system shares an adjacent pair of atoms with other rings in the system, wherein one or more rings may contain one or more double bonds, but none of them has a fully conjugated π-electron system, wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O)r (wherein r is an integer of 0, 1, 2), the remaining ring atoms are carbon. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocycloalkyl, preferably “3-12-membered bicyclic fused heterocycloalkyl” containing 1 to 3 heteroatoms selected from N, O or S; more preferably saturated “3-12-membered bicyclic fused heterocycloalkyl” containing 1 or 2 N atoms. Non-limiting examples of fused heterocycloalkyl include:

##STR00399##

[1322] “Bridged heterocycloalkyl” refers to a polycyclic heterocycloalkyl in which any two rings share two atoms that are not directly connected. They can contain one or more double bonds, but no ring has a fully conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O)r (wherein r is an integer of 0, 1, 2), and the remaining ring atoms are carbon. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl. The non-limiting examples of bridged heterocycloalkyl include:

##STR00400##

[1323] The heterocycloalkyl ring can be fused to the aryl, heteroaryl or cycloalkyl, wherein the ring connected with the parent structure is heterocycloalkyl, non-limiting examples include:

##STR00401##

the heterocycloalkyl may be optionally substituted or unsubstituted.

[1324] “Aryl” refers to all-carbon monocyclyls or fused polycyclyls (that is, rings sharing adjacent pairs of carbon atoms) and polycyclyls having conjugated π-electron systems, and “6-10-membered aryl” refers to all-carbon aryls containing 6-10 carbons, such as phenyl and naphthyl: preferably phenyl. The aryl ring may be fused to the heteroaryl, heterocycloalkyl or cycloalkyl, wherein the ring connected with the parent structure is an aryl ring, and the non-limiting examples include:

##STR00402##

the aryl may be optionally substituted or unsubstituted.

[1325] “Heteroaryl” refers to a heteroaromatic system comprising 1 to 4 heteroatoms, the heteroatoms include nitrogen, oxygen or S(O)r (wherein r is an integer of 0, 1, 2), 5-6-membered heteroaryl refers to a heteroaromatic system containing 5 to 6 ring atoms, and 5-10-membered heteroaryl refers to a heteroaromatic system containing 5 to 10 ring atoms, preferably 5-6-membered heteroaryl; more preferably a 5-6-membered heteroaryl containing 1 or 2 N atoms; non-limiting examples include furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidyl, pyrazinyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, etc.; preferably pyridyl. The heteroaryl ring may be fused to the aryl, heterocycloalkyl or cycloalkyl, wherein the ring connected with the parent structure is a heteroaryl, and the non-limiting examples include:

##STR00403##

the heteroaryl may be optionally substituted or unsubstituted.

[1326] “Alkenyl” means an alkyl as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, and “C2-8 alkenyl” means a linear or branched alkenyl containing 2-8 carbons, including but not limited to vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc., preferably “C2-6 alkenyl”, more preferably “C2-4 alkenyl”. The alkenyl may be substituted or unsubstituted.

[1327] “Alkynyl” refers to an alkyl as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, and “C2-8 alkynyl” refers to a linear or branched alkynyl containing 2-8 carbons, including but not limited to ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, preferably “C2-6 alkynyl”, more preferably “C2-4 alkynyl”. The alkynyl may be substituted or unsubstituted.

[1328] “Subgroup” refers to a divalent, such as alkylene refers to divalent alkyl, alkenylene refers to divalent alkenyl, alkynylene refers to divalent alkynyl, cycloalkylene to divalent cycloalkyl, heterocycloalkylene to divalent heterocycloalkyl, arylene to divalent aryl, heteroarylene to divalent heteroaryl, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl as defined above, the subgroups may be optionally substituted or unsubstituted.

[1329] “Alkoxy” refers to —O-(alkyl), wherein the alkyl is as defined above. “C1-C8 alkoxy” refers to an alkyloxy containing 1 to 8 carbons. Non-limiting examples include methoxy, ethoxy, propoxy, butoxy, etc. The alkoxy may be optionally substituted or unsubstituted.

[1330] “Cycloalkoxy” refers to —O-(unsubstituted cycloalkyl), wherein the cycloalkyl is as defined above. C3-C8 cycloalkoxy refers to cycloalkyloxys containing 3 to 8 carbons. Non-limiting examples include cyclopropoxy, cyclobutoxy, cyclopentoxy, cyclohexoxy, etc. The cycloalkoxy may be optionally substituted or unsubstituted.

[1331] “Haloalkyl” refers to an alkyl substituted by one or more fluorine, chlorine, bromine or iodine atoms, wherein the alkyl is as defined above, and non-limiting examples include difluoromethyl, dichloromethyl, dibromomethyl, trifluoromethyl, trichloromethyl, tribromomethyl, etc.

[1332] “Haloalkoxy” refers to a group in which hydrogen on an alkyl is substituted by one or more fluorine, chlorine, bromine or iodine atoms, wherein the alkoxy is as defined above. For example, difluoromethoxy, dichloromethoxy, dibromomethoxy, trifluoromethoxy, trichloromethoxy, tribromomethoxy, etc.

[1333] “Hydroxyalkyl” refers to an alkyl optionally substituted by one or more —OH, wherein the alkyl is as defined above, and non-limiting examples include hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxyisopropyl.

[1334] “Aminoalkyl” refers to an alkyl optionally substituted by one or more —NH.sub.2, wherein the alkyl is as defined above, and non-limiting examples include aminomethyl, aminoethyl, aminopropyl, aminoisopropyl.

[1335] “Amino” and “amine” are recognized in the art and refer to substituted or unsubstituted ammonia.

[1336] “Carboxyl” or “carboxylic acid” refers to —COOH.

[1337] “Oxo” refers to ═O.

[1338] “Halogen” refers to fluorine, chlorine, bromine or iodine.

[1339] “Cyano” refers to —CN.

[1340] “Hydroxyl” refers to —OH.

[1341] “Raney-Ni” refers to Raney nickel (hydrogen active catalytic reducing agent).

[1342] “Dess-martin” refers to 1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3-(1H)-one.

[1343] “PdCl.sub.2(dppf)” refers to [1,1-bis (diphenylphosphorus) ferrocene] palladium dichloride.

[1344] “T3P” refers to 2,4, 6-tripropyl-1,3,5,2,4,6-trioxytriphosphate-2,4,6-trioxide.

[1345] “HATU” refers to 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate.

[1346] “DPBS” refers to Dulbecco's phosphate buffer saline.

[1347] “PBS” refers to phosphate buffer saline.

[1348] “SDS-PAGE” refers to sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

[1349] “PVDF” refers to polyvinylidene fluoride.

[1350] “More” independently refers to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[1351] “Optional” means that a subsequently described event or environment may, but does not have to, occur, and that description includes the place where the event or environment occurs or does not occur. For example, “a heterocycloalkyl optionally substituted by an alkyl” means that an alkyl may, but does not have to be present, and the description includes the case where the heterocycloalkyl is substituted by an alkyl and the case where the heterocycloalkyl is not substituted by an alkyl.

[1352] “Substituted” means that one or more hydrogen atoms, preferably at most 5, more preferably 1 to 3 hydrogen atoms are substituted independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (experimentally or theoretically) possible or impossible substitutions without too much effort. For example, amino or hydroxyls with free hydrogen may be unstable when combined with carbon atoms with unsaturated (e. g. olefinic) bonds.

[1353] “Pharmaceutical composition” means a mixture comprising one or more compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs with other chemical components, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to promote the administration of organisms, facilitate the absorption of active ingredients and thus exert biological activity.

[1354] The present invention also provides pharmaceutically acceptable salts of compounds of formula (I). The term “pharmaceutically acceptable salt” refers to an acid addition salt or a base addition salt of a compound of the present invention that is relatively non-toxic. The acid addition salts are salts of the compounds of formula (I) of the present invention and suitable inorganic or organic acids, these salts can be prepared during the final separation and purification of the compounds, or can be prepared by reacting the purified compounds of formula (I) in their free base form with suitable organic or inorganic acids. Representative acid addition salts include hydrobromate, hydrochloride, sulfate, bisulfate, sulfite, acetate, oxalate, valerate, oleate, palmitic acid, stearate, lauroleate, borate, benzoate, lactate, phosphate, hydrophosphate, carbonate, bicarbonate, toluate, citrate, maleate, fumarate, succinate, tartrate, benzoate, mesylate, p-toluene sulfonate, gluconate, lactobionate and lauryl sulfonate, etc. The base addition salt is a salt formed by a compound of formula (I) and a suitable inorganic or organic base, including, for example, a salt formed with alkali metals, alkaline earth metals, and quaternary ammonium cations, such as sodium salt, lithium salt, potassium salt, calcium salt, magnesium salt, tetramethylquaternary ammonium salt, tetraethyl quaternary ammonium salt, etc.; amine salt includes salts formed with ammonia (NH.sub.3), primary amine, secondary amine or tertiary amine, such as methylamine salt, dimethylamine salt, trimethylamine salt, triethylamine salt, ethylamine salt, etc.

[1355] The compounds of the invention, or pharmaceutically acceptable salts thereof, can be administered to mammals, including humans, orally, rectally, parenterally (intravenous, intramuscular or subcutaneous), topically (powder, ointment or drops), or intratumorally.

[1356] The dosage of the compound of the present invention may be about 0.05-300 mg/kg body weight/day, preferably 10-300 mg/kg body weight/day, more preferably 10-150 mg/kg body weight/day.

[1357] The compounds of the present invention, or pharmaceutically acceptable salts thereof, may be formulated as solid dosage forms for oral administration, including, but not limited to, capsules, tablets, pills, powders, granules, and the like. In these solid dosage forms, the active ingredient, i.e. the compound of formula (I) of the present invention, is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or mixed with any of the following components: (1) fillers or compatibilizer, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid, etc.; (2) binders, for example, hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and arabic gum, etc.; (3) humectants, for example, glycerol, etc; (4) disintegrating agents, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain composite silicates, and sodium carbonate, etc; (5) dissolution-retarding agents, for example, paraffin, etc; (6) absorption accelerators, for example, quaternary ammonium compounds, etc; (7) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (8) adsorbents, for example, kaolin, etc; and (9) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or the mixture thereof. Buffering agents may also be included in capsules, tablets and pills.

[1358] The solid dosage forms such as tablets, sugar pills, capsules, pills and granules can be coated or microencapsulated with coating and shell materials such as enteric coatings and other materials known in the art. They may contain opaque agents, and the release of the active ingredient in such a composition may be released in a delayed manner in a part of the digestive tract. Examples of embedding components that can be employed are polymeric materials and waxy materials. If necessary, the active ingredient may also form a microcapsule form with one or more of the excipients described above.

[1359] The compounds of the present invention, or pharmaceutically acceptable salts thereof, may be formulated as liquid dosage forms for oral administration, including, but not limited to, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, tinctures, and the like. In addition to the compounds of formula (I) or pharmaceutically acceptable salts thereof as active ingredients, liquid dosage forms may contain inert diluents such as water and other solvents, solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or a mixture of these substances as conventionally used in the art. In addition to these inert diluents, the liquid dosage forms of the present invention may also contain conventional auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and spices.

[1360] The suspending agent includes, for example, ethoxylated isooctadecanol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methanol and agar, or a mixture of these substances.

[1361] The compounds of the present invention, or pharmaceutically acceptable salts thereof, may be formulated as dosage forms for parenteral injection including, but not limited to, physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for re-dissolution into sterile injectable solutions or dispersions. Suitable carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[1362] The compounds of the present invention, or pharmaceutically acceptable salts thereof, may also be formulated as dosage forms for topical administration, including, for example, ointments, powders, suppositories, drops, propellants, inhalants, and the like. The compounds of formula (I) of the invention, or pharmaceutically acceptable salts thereof, as active ingredient, are mixed with a physiologically acceptable carrier and optionally a preservative, a buffer, or, if necessary, a propellant as may be required under sterile conditions.

[1363] The present invention also provides a pharmaceutical composition comprising a compound of formula (I) of the present invention or a pharmaceutically acceptable salt thereof as active ingredient, and a pharmaceutically acceptable carrier, excipient or diluent. In the preparation of pharmaceutical compositions, a compound of formula (I) or a pharmaceutically acceptable salt thereof is usually mixed with a pharmaceutically acceptable carrier, excipient or diluent.

[1364] The composition of the present invention can be formulated as a conventional pharmaceutical preparation according to the conventional preparation method. For example, tablets, pills, capsules, powders, granules, emulsions, suspensions, dispersions, solutions, syrups, elixirs, ointments, drops, suppositories, inhalants, propellants, etc.

[1365] The compound of the present invention or its pharmaceutically acceptable salt may be administered alone, or (if necessary) in combination with other pharmaceutically acceptable therapeutic agents, such as in combination with other anti-tumor drugs, anti-inflammatory drugs or autoimmune drugs. The ingredients to be combined may be administered simultaneously or sequentially, in the form of a single formulation or in the form of a different formulation. The combination may include not only a combination of a compound of the present invention and one other active agent, but also a combination of a compound of the present invention and two or more other active agents.

[1366] The present invention proves that the compound of formula I of the present invention can effectively bind to the IRAK4 target protein or produce an inhibitory effect through the IRAK4 kinase activity test experiment, and the compound of formula I of the present invention can effectively and specifically degrade the IRAK4 protein in THP-1 cells by Western-Blot. It is proved by AlphaLISA detection method that the compound of formula I of the present invention can effectively inhibit the production of IL-6 by immune cells, and the compound of the present invention has good degradation selectivity, and the compound of formula I of the present invention, and/or its stereoisomers, enantiomers, diastereomers, deuterates, hydrates, solvates, metabolites, prodrugs and/or pharmaceutically acceptable salts can effectively and selectively degrade IRAK4 protein, so as to achieve the effect of preventing or treating diseases or conditions related to IRAK4.

EXAMPLES

[1367] Hereinafter, the present invention will be described in further detail and completely with reference to the examples, but the present invention is not in any way limited to the contents of the examples. The starting materials in the examples of the present invention are known and can be commercially available or can be synthesized using or in accordance with methods known in the art. In the absence of special instructions, in the present examples, the experimental method that does not specify the specific conditions, usually in accordance with the conventional conditions, or in accordance with the conditions suggested by raw material or commodity manufacturer.

I Examples for Compounds Preparation

Intermediate 1: 2-Fluoro-4-methoxy-5-nitrobenzaldehyde

[1368] ##STR00404##

[1369] Cesium carbonate (3.5 g, 10.8 mmol) and potassium iodide (1.85 g, 13.0 mmol) were added to a solution of 2-fluoro-4-hydroxy-5-nitrobenzaldehyde (2 g, 10.8 mmol) in N,N-dimethylformamide (20 ml). The reaction solution was stirred for 10 hours at room temperature, water (30 ml) was added, extracted with ethyl acetate for three times, the organic phase was collected, washed with saturated saline, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the concentrate was purified by column to obtain 1.6 g of the target product 2-fluoro-4 methoxy-5-nitrobenzaldehyde.

[1370] LC-MS: (ES, m/z): [M+H].sup.+=200.1

Intermediate 2: 2-Fluoro-5-nitro-4-trifluoromethoxy) benzaldehyde

[1371] ##STR00405##

[1372] Nitric acid (1.0 mL, 65%) was added dropwise to a solution of 2-fluoro-4-(trifluoromethoxy) benzaldehyde (1.0 g, 4.8 mmol) in sulfuric acid (5 ml) while keeping the temperature below −10° C. After dripping, the solution was poured into ice water (20 mL) and stirred for 10 minutes. Filtered, solid was washed with water (10 mL×3), and dried under reduced pressure to obtain 0.9 g of crude target product. The crude product can be used for the next step without further purification.

[1373] LC-MS: (ES, m/z): [M+H].sup.+=254.0

Intermediate 3: N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: Preparation of 2-azido-4-methoxy-5-nitrobenzaldehyde

[1374] ##STR00406##

[1375] Sodium azide (1.06 g, 16.1 mmol) was added to a solution of 2-fluoro-4 methoxy-5-nitrobenzaldehyde (1.6 g, 8.04 mmol) in dimethyl sulfoxide (20 ml), stirred at room temperature for 2 hours, then the reaction solution was poured into ice water, extracted with ethyl acetate, the organic phase was collected, washed with saturated saline, dried over anhydrous sodium sulfate, and 1.7 g crude product was obtained by concentration, which was directly used in the next reaction without purification.

Step 2: Preparation of tert-butyl 4-(6-methoxy-5-nitro-2H-indazol-2-yl) piperidine-1-carboxylate

[1376] ##STR00407##

[1377] The solution of 2-azido-4-methoxy-5-nitrobenzaldehyde (1.7 g, 7.65 mmol) and tert-butyl 4-aminopiperidine-1-carboxylate (1.53 g, 7.65 mmol) in toluene (20 ml) was stirred and reacted at 100° C. for 2 hours, cooled to room temperature, the reaction solution was concentrated, 50 ml of water was added, extracted with ethyl acetate, the organic phase was collected, washed with saturated saline, dried over anhydrous sodium sulfate, the organic phase was concentrated, and the concentrate was purified by column to obtain 1.72 g of tert-butyl 4-(6-methoxy-5-nitro-2H-indazol-2-yl) piperidin-1-carboxylate.

[1378] LC-MS: (ES, m/z): [M+H].sup.+=377.2

Step 3: Preparation of tert-butyl 4-(5-amino-6-methoxy-2H-indazol-2-yl) piperidine-1-carboxylate

[1379] ##STR00408##

[1380] Iron powder (2.85 g, 50.9 mmol) and ammonium chloride (0.13 g, 2.3 mmol) were added to a solution of tert-butyl 4-(6-methoxy-5-nitro-2H-indazol-2-yl) piperidin-1-carboxylate (1.7 g, 4.5 mmol) in ethanol (20 ml), and the reaction solution was reacted at 90° C. for 2 hours. After the reaction solution was cooled to room temperature, it was filtered and concentrated to obtain 2.01 g of crude target product, which was directly used for the next reaction.

[1381] LC-MS: (ES, m/z): [M+H].sup.+=347.3

Step 4: Preparation of tert-butyl 4-(6-methoxy-5-(6-(trifluoromethyl) pyridine-2-amido)-2H-indazol-2-yl) piperidine-1-carboxylate

[1382] ##STR00409##

[1383] 6-(Trifluoromethyl) pyridine-2-carboxylic acid (940 mg, 4.9 mmol) and N,N-diisopropylethylamine (1.9 g, 14.7 mmol) were added to a solution of tert-butyl 4-(5-amino-6-methoxy-2H-indazol-2-yl) piperidine-1-carboxylate (1.7 g, 4.9 mmol) in tetrahydrofuran, then T.sub.3P (1.5 g) was added, and the reaction solution was stirred for 2 hours at room temperature. After concentrating the reaction solution, water was added, extracted with ethyl acetate for three times, and the organic phase was collected, washed with saturated saline, dried over anhydrous sodium sulfate and concentrated. The concentrate was purified by column to obtain 1.78 g of the target product tert-butyl 4-(6-methoxy-5-(6-(trifluoromethyl) pyridine-2-amide)-2H-indazol-2-yl) piperidine-1-carboxylate.

[1384] LC-MS: (ES, m/z): [M+H].sup.+=520.2

Step 5: Preparation of N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1385] ##STR00410##

[1386] Trifluoroacetic acid (1 ml) was added to a solution of tert-butyl 4-(6-methoxy-5-(6-(trifluoromethyl) pyridine-2-amido)-2H-indazol-2-yl) piperidine-1-carboxylate (435 mg, 0.795 mmol) in dichloromethane. The reaction solution was stirred at room temperature for 1h. The solvent was removed by concentration under reduced pressure to obtain the target crude product (500 mg). The crude product was used directly for the next step.

[1387] LC-MS: (ES, m/z): [M+H].sup.+=420.2

[1388] The following intermediates were prepared with reference to the method of intermediate 3.

TABLE-US-00001 Intermediate Structure Name LC-MS Intermediate 4 [00411] N-(2-(piperidin-4-yl)-6-(trifluoro- methoxy)-2H-indazol-5-yl)-6-(trifluo- romethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 474.1.

Intermediate 5: N-(6-(2-hydroxyprop-2-yl)-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: Preparation of methyl 5-nitro-1H-indazole-6-carboxylate

[1389] ##STR00412##

[1390] At −10° C., concentrated nitric acid (15 mL) was slowly added to a solution of methyl 1H-indazole-6-carboxylate (9.2 g, 0.052 mol) in concentrated sulfuric acid (20 mL). The reaction solution was stirred at −10° C. for 2h. The reaction solution was poured into ice water, and the solid was filtered to obtain 10.8 g of the target crude product as a white solid. The crude product was used directly for the next step.

[1391] LC-MS: (ES, m/z): [M+H].sup.+=222.1

Step 2: Preparation of methyl 2-(1-(tert-butoxycarbonyl) piperidin-4-yl)-5-nitro-2H-indazole-6-carboxylate

[1392] ##STR00413##

[1393] Potassium carbonate (11.1 g, 81.3 mmol) was added to a solution of methyl 5-nitro-1H-indazol-6-carboxylate (6 g, 27.12 mmol) and tert-butyl 4-(p-toluenesuloyloxy) piperidine-1-carboxylate (15.3 g, 43.5 mmol) in N,N-dimethylformamide (100 mL). The reaction solution was stirred at 100° C. for 10h. The reaction solution was cooled to room temperature, water (50 ml) and ethyl acetate (100 ml) were added, the water layer was separated and extracted with ethyl acetate (100 ml×2). The organic layers were combined, washed with saline (100 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 2.01 g of target product as a yellow solid.

[1394] LC-MS: (ES, m/z): [M+H].sup.+=405.2

Step 3: Preparation of methyl 5-amino-2-(1-(tert-butoxycarbonyl) piperidin-4-yl)-2H-indazole-6-carboxylate

[1395] ##STR00414##

[1396] Iron powder (2.85 g, 50.9 mmol) and ammonium chloride (0.13 g, 2.3 mmol) were added to a solution of methyl 2-(1-(tert-butoxycarbonyl) piperidin-4-yl)-5-nitro-2H-indazole-6-carboxylate (1.7 g, 4.5 mmol) in ethanol (20 ml). The reaction solution was stirred at 90° C. for 2h. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain 2.01 g of the crude target product as an oil. The crude product was used directly for the next step.

[1397] LC-MS: (ES, m/z): [M+H].sup.+=375.3

Step 4: methyl 2-(1-(tert-butoxycarbonyl) piperidin-4-yl)-5-(6-(trifluoromethyl) pyridine carboxamide)-2H-indazole-6-carboxylate

[1398] ##STR00415##

[1399] T.sub.3P (650 mg) was added to a mixture of methyl 5-amino-2-(1-(tert-butoxycarbonyl) piperidin-4-yl)-2H-indazole-6-carboxylate (170 mg, 0.45 mmol), 6-(trifluoromethyl) pyridine-2-carboxylic acid (86.8 mg, 0.45 mmol) and N,N-diisopropylethylamine (88 mg, 0.68 mmol) in tetrahydrofuran. The reaction solution was stirred at room temperature for 2 hours. The solvent was removed by concentration under reduced pressure. Water (20 mL) and ethyl acetate (20 mL) were added. The water layer was separated and extracted with ethyl acetate (20 ml×2). The organic layers were combined, washed with saline (20 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 150 mg of target product as a yellow solid.

[1400] LC-MS: (ES, m/z): [M+H].sup.+=548.3

Step 5: Preparation of tert-butyl 4-(6-(2-hydroxyprop-2-yl)-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidine-1-carboxylate

[1401] ##STR00416##

[1402] Lithium chloride (192 mg, 4.57 mmol) was added to a solution of methyl 2-(1-(tert-butoxycarbonyl) piperidin-4-yl)-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazole-6-carboxylate (500 mg, 0.91 mmol) in tetrahydrofuran. The reaction solution was cooled to 0° C., and then tetrahydrofuran methylmagnesium bromide (3.04 ml, 1M) was added. The reaction solution was stirred at room temperature for 12h, quenched with ammonium chloride aqueous solution, and water (30 mL) and ethyl acetate (50 mL) were added. The water layers were separated and extracted with ethyl acetate (50 ml×2), the organic layers were combined, washed with saline (50 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 435 mg of target product as a white solid.

[1403] LC-MS: (ES, m/z): [M+H].sup.+=548.3

Step 6: Preparation of N-(6-(2-hydroxyprop-2-yl)-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1404] ##STR00417##

[1405] A mixture of tert-butyl 4-(6-(2-hydroxyprop-2-yl)-5-(6-(trifluoromethyl) pyridineformamido)-2H-indazol-2-yl) piperidine-1-carboxylate (435 mg, 0.79 mmol) in dichloromethane (3 mL) and trifluoroacetic acid (1 mL) was stirred and reacted at room temperature for 1 h, and the solvent was removed by concentration under reduced pressure, 500 mg of target crude product was obtained as a yellow oil. The crude product was used directly for the next step.

[1406] LC-MS: (ES, m/z): [M+H].sup.+=448.2

Intermediate 6: N-(6-(2-hydroxyprop-2-yl)-1H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: methyl 5-amino-1H-indazole-6-carboxylate

[1407] ##STR00418##

[1408] Palladium (800 mg, 10% w.t) was added to a solution of methyl 5-nitro-1H-indazole-6-carboxylate (5.37 g, 0.024 mol) in ethyl acetate (500 mL). The reaction mixture was stirred for 16 hours at room temperature in a hydrogen atmosphere. The solids were filtered and washed with ethyl acetate (100 mL×2). The filtrate was concentrated under reduced pressure to obtain 4.45 g of crude target product as a brown solid. The crude product was used directly for the next reaction.

[1409] LC-MS: (ES, m/z): [M+H].sup.+=192.2.

Step 2: methyl 5-(6-(trifluoromethyl) pyridine carboxamide)-1H-indazole-6-carboxylate

[1410] ##STR00419##

[1411] N,N-diisopropylethylamine (4.5 g, 0.0345 mol) was added to a solution of methyl 5-amino-1H-indazole-6-carboxylate (4.4 g, 0.023 mol) and 6-(trifluoromethyl) picolinic acid (4.4 g, 0.023 mol) in tetrahydrofuran (50 ml), the mixture was cooled to 0° C., and then T3P (18.0 g, 0.0345 mol) was added in batches, and the reaction mixture was stirred at room temperature for 2 hours. The solvent was removed by concentration under reduced pressure, water (50 ml) and ethyl acetate (100 ml) were added, and the water layer was separated and extracted with ethyl acetate (100 ml×2). The organic layer was collected, washed with saturated saline (100 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 7.87 g of target product as a yellow solid.

[1412] LC-MS: (ES, m/z): [M+H].sup.+=365.1.

Step 3: N-(6-(2-hydroxyprop-2-yl)-1H-indazol-5-yl)-6-(trifluoromethyl) pyridine carboxamide

[1413] ##STR00420##

[1414] The solution of methyl 5-(6-(trifluoromethyl) pyridinecarboxamido)-1H-indazole-6-carboxylate (1 g, 0.002 mol) and lithium chloride (576 mg, 0.013 mol) in tetrahydrofuran (30 mL) was cooled to −10° C., and then methylmagnesium bromide (9.1 mL, 0.027 mol) was added under nitrogen protection. After the reaction mixture was stirred at room temperature for 8 hours, it was cooled to 0° C., and saturated ammonium chloride aqueous solution (50 mL), water (50 ml) and ethyl acetate (100 ml) were added to quench the solution. The water layer was separated and extracted with ethyl acetate (100 ml×2). The organic layer was collected, washed with saturated salt water (100 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 532 mg of target product as a yellow solid.

[1415] LC-MS: (ES, m/z): [M+H].sup.+=365.1.

Intermediate 7: 3-(2, 4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoic acid

Step 1: Preparation of 3-(2,4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoic acid

[1416] ##STR00421##

[1417] 3-Amino-4-methoxybenzoic acid (5.0 g, 2.93 mmol) was suspended in acrylic acid (8.05 mL, 117 mmol) and stirred at 100° C. for 3 h, then the reaction solution was stirred and cooled to room temperature. Acetic acid (33 ml) was added, the stirred suspension was heated at 100° C. for 10 minutes, urea (11.00 g, 183 mmol) was added, and the reaction solution was stirred at 120° C. overnight. The reaction solution was added to a mixture of ice water and concentrated hydrochloric acid (37%), stirred, and the obtained suspension was stored overnight in a refrigerator at 5° C., then filtered, and the solid was washed with water and dried to obtain the solid. The solid was ground in hydrochloric acid solution (0.05M), filtered, washed with methyl tert-butyl ether, and dried under reduced pressure at 40° C. to obtain 6.29 g of target product.

[1418] .sup.1H NMR (400 MHz, DMSO) δ 12.70 (s, 1H), 10.34 (s, 1H), 7.92 (dd, J=8.6, 2.2 Hz, 1H), 7.83 (d, J=2.2 Hz, 1H), 7.21 (d, J=8.8 Hz, 1H), 3.94-3.82 (m, 3H), 3.60 (t, J=6.7 Hz, 2H), 2.69 (s, 2H).

[1419] The following intermediates were prepared with reference to the method of intermediate 7:

TABLE-US-00002 Intermediate Structure Name LC-MS Intermediate 8 [00422] 3-(2,4- dioxotetrahydropyrimi- din- 1(2H)-yl)-4-ethoxy- benzoic acid (ES, m/z): [M + H].sup.+ = 279.1 Intermediate 9 [00423] 4-Chloro-3-(2,4- dioxotetrahydropyrimi- din-1(2H)-yl)benzoic acid (ES, m/z): [M + H].sup.+ = 269.0 Intermediate 10 [00424] 3-(2,4- dioxotetrahydropyrimi- din-1(2H)- yl)-4-(trifluorometh- oxy)benzoic acid (ES, m/z): [M + H].sup.+ = 319.1 Intermediate 11 [00425] 3-(2,4- dioxotetrahydropyrimi- din-1(2H)- yl)-4-fluorobenzoic acid (ES, m/z): [M + H].sup.+ = 253.0 Intermediate 12 [00426] 3-(2,4- dioxotetrahydropyrimi- din-1(2H)- yl)-4-(trifluorometh- yl)benzoic acid (ES, m/z): [M + H].sup.+ = 303.0 Intermediate 13 [00427] 3-(2,4- dioxotetrahydropyrimi- din-1(2H)-yl)benzoic acid (ES, m/z): [M + H].sup.+ = 235.1

Intermediate 14: 3-(2, 6-dioxopiperidin-3-yl)-4-fluorobenzoic acid

Step 1: Preparation of 2, 6-bis (benzyloxy) pyridine

[1420] ##STR00428##

[1421] Benzyl alcohol (109.6 g, 1.01 mol) was added to a solution of sodium hydride (60% mineral oil mixture, 54.0 g, 1.35 mol) in N,N-dimethylformamide (600 mL) at 0° C. After stirring for 30 minutes, 2,6-dichloropyridine (50.0 g, 0.338 mol) was added. The reaction mixture was heated to 80° C. and stirred overnight, and after cooling to room temperature, the reaction mixture was quenched with ice water (1000 mL). The solid was filtered and dried to obtain 75.9 g of target product.

[1422] .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ 7.68 (t, J=8.4 Hz, 1H), 7.48-7.36 (m, 10H), 6.45 (d, J=8.4 Hz, 2H), 5.38 (s, 4H).

[1423] LC-MS: (ES, m/z): [M+H].sup.+=292.2.

Step 2: Preparation of 2,6-bis (benzyloxy)-3-bromopyridine

[1424] ##STR00429##

[1425] N-bromosuccinimide (41.7 g, 0.23 mol) was added to a solution of 2,6-bis (benzyloxy) pyridine (75.9 g, 0.26 mol) in acetonitrile (600 mL). The reaction mixture was stirred at 80° C. for 2 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The concentrate was diluted with ethyl acetate (500 mL) and washed with water and saturated saline. The organic layer was collected and dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 75.0 g of target product.

[1426] .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ 7.88 (d, J=8.4 Hz, 1H), 7.45-7.37 (m, 10H), 6.44 (d, J=8.4 Hz, 1H), 5.42 (s, 2H), 5.33 (s, 2H).

[1427] LC-MS: (ES, m/z): [M+H].sup.+=370.1.

Step 3: Preparation of 3-(2, 6-bis (benzyloxy) pyridin-3-yl)-4-fluorobenzoic acid

[1428] ##STR00430##

[1429] PdCl.sub.2(dppf)-chloroform complex (612 mg, 0.84 mmol) and sodium carbonate (1.77 g, 16.74 mmol) were added to a mixture of 2, 6-bis (benzyloxy)-3-bromopyridine (3.1 g, 8.37 mmol) and 3-boro-4-fluorobenzoic acid (1.85 g, 10.05 mmol) in dioxane (31 mL) and water (7.5 mL). The reaction mixture was heated to 90° C. under nitrogen protection and the reaction was maintained for 16 hours. After the reaction was completed, 15% hydrochloric acid solution (20 mL) was added, extracted with ethyl acetate (100 mL×3), washed with saturated saline (50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 1.50 g of target product.

[1430] LC-MS: (ES, m/z): [M+H].sup.+=430.2

Step 4: Preparation of 3-(2, 6-dioxopiperidin-3-yl)-4-fluorobenzoic acid

[1431] ##STR00431##

[1432] Palladium/carbon (10%, 120 mg) was added to a solution of 3-(2,6-bis (benzyloxy) pyridin-3-yl)-4-fluorobenzoic acid (1.50 g, 3.49 mmol) in methanol (10 mL). At room temperature, the reaction mixture was stirred in a hydrogen atmosphere for 48 hours. After the reaction was completed, the reaction mixture was filtered through diatomite, and the filtrate was concentrated under reduced pressure. The concentrate was purified to obtain 600 mg of target product.

[1433] LC-MS: (ES, m/z): [M+H].sup.+=252.1

[1434] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.04 (s, 1H), 10.92 (s, 1H), 7.91-7.95 (m, 2H), 7.30-7.34 (m, 1H), 4.15-4.19 (m, 1H), 2.70-2.79 (m, 1H), 2.53-2.57 (m, 1H), 2.23-2.27 (m, 1H), 2.01-2.07 (m, 1H).

Intermediate 15: pentafluorophenyl 3-(2, 4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoate

[1435] ##STR00432##

[1436] A mixture of 3-(2, 4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoic acid (2.0 g, 7.58 mmol), 2,3,4,5, 6-pentafluorophenol (1.67 g, 9.09 mmol) and N,N′-dicyclohexylcarbimide (1.87 g, 9.09 mmol) in N,N-dimethylformamide (20 mL) was stirred for 3h at room temperature. The reaction solution was poured into water (200 mL) and stirred for 0.5h. The solution was extracted with ethyl acetate (3×200 mL). The organic phase was collected, washed with water (2×500 mL) and saturated saline (300 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 3.0 g of target product as a brown solid.

[1437] LC-MS: (ESI, m/z): [M+H].sup.+=431.1

[1438] The following intermediates were prepared with reference to the method of intermediate 15:

TABLE-US-00003 Intermediate Structure Name LC-MS Intermediate 16 [00433] pentafluorophenyl 4-chloro-3-(2,4- dioxotetrahydropyrimi- din-1(2H)-yl) benzoate (ES, m/z): [M + H].sup.+ = 435.0 Intermediate 17 [00434] pentafluorophenyl 3-(2,4- dioxotetrahydropyrimi- din-1 (2H)-yl)-4-ethoxy- benzoate (ES, m/z): [M + H].sup.+ = 445.1 Intermediate 18 [00435] pentafluorophenyl 3-(2,4- dioxotetrahydropyrimi- din-1 (2H)-yl)-4-(trifluorometh- oxy)benzoate (ES, m/z): [M + H].sup.+ = 485.0 Intermediate 19 [00436] pentafluorophenyl 3-(2,4- dioxotetrahydropyrimi- din-1 (2H)-yl)-4-fluorobenzo- ate (ES, m/z): [M + H].sup.+ = 419.0 Intermediate 20 [00437] pentafluorophenyl 3-(2,4- dioxotetrahydropyrimi- din-1 (2H)-yl)-4-(trifluorometh- yl)benzoate (ES, m/z): [M + H].sup.+ = 469.0 Intermediate 21 [00438] pentafluorophenyl 3-(2,4- dioxotetrahydropyrimi- din-1(2H)-yl) benzoate (ES, m/z): [M + H].sup.+ = 401.0 Intermediate 22 [00439] pentafluorophenyl 3-(2,6- dioxopiperidin-3-yl)- 4-fluorobenzoate (ES, m/z): [M + H ].sup.+ = 418.0

Intermediate 23: 1-(2-methoxy-5-(3, 9-diazaspiro[5.5] undecyl-3-carbonyl) phenyl) dihydropyrimidine-2, 4(1H,3H)-dione

Step 1: Preparation of tert-butyl 9-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-3, 9-diazaspiro[5.5] undecyl-3-carboxylate

[1439] ##STR00440##

[1440] At room temperature, under the protection of nitrogen, HATU (1.66 g, 4.37 mmol), tert-butyl 3, 9-diazaspiro[5.5] undecyl-3-carboxylate (0.96 g, 3.78 mmol) and N-methylmorpholine (0.8 g, 7.92 mmol) were added to a solution of 3-(2, 4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoic acid (1.0 g, 3.78 mmol) in N,N-dimethylformamide (10 mL). After stirring for 2h, the reaction mixture was quenched with water (50 mL) and extracted with ethyl acetate (3×50 mL). The organic layer was collected and washed with saturated saline (3×50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 1.55 g of target product as a white solid.

[1441] LC-MS: (ES, m/z): [M+H].sup.+=501.2

Step 2: Preparation of 1-(2-methoxy-5-(3, 9-diazaspiro [5.5] undecyl-3-carbonyl) phenyl) dihydropyrimidine-2,4 (1H,3H)-dione

[1442] ##STR00441##

[1443] The reaction solution of tert-butyl 9-(3-(2, 4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoyl)-3, 9-diazaspiro[5.5] undecyl-3-carboxylate (1.5 g, 3.1 mmol) in trifluoroacetic acid/dichloromethane (2.5 mL/5 mL) was stirred at room temperature for 4h. The reaction solution was concentrated under reduced pressure to obtain 2.7 g of crude target compound as a yellow oil. The crude product was used directly for the next reaction.

[1444] LC-MS: (ESI, m/z): [M+H].sup.+=401.1

[1445] The following intermediates were prepared according to the method of intermediate 23:

TABLE-US-00004 Intermediate Structure Name LC-MS Intermediate 24 [00442] 1-(2-methoxy-5-(2,7- diazaspiro[3.5] nonyl-7-carbonyl) phenyl) dihydropyrimidine- 2,4(1H,3H)-dione (ES, m/z): [M + H].sup.+ = 373.2 Intermediate 25 [00443] 1-(2-methoxy-5-(2,7- diazaspiro[3.5] nonyl-2-carbonyl) phenyl) dihydropyrimidine- 2,4(1H,3H)-dione (ES, m/z): [M + H].sup.+ = 373.2 Intermediate 26 [00444] 1-(2-chloro-5-(3,9- diazaspiro[5.5]un- decyl-3-carbonyl) phenyl) dihydropyrimidine- 2,4(1H,3H)-dione (ES, m/z): [M + H].sup.+ = 405.1

Intermediate 27: 2-(9-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) acetaldehyde

Step 1: Preparation of 1-(5-(9-(2, 2-dimethoxyethyl)-3, 9-diazaspiro[5.5]undecyl-3-carbonyl)-2-methoxyphenyl) dihydropyrimidine-2,4(1H,3H)-dione

[1446] ##STR00445##

[1447] 2-Bromo-1, 1-dimethoxyethane (46 mg, 0.3 mmol) was added to a mixture of 1-(2-methoxy-5-(3, 9-diazaspiro [5.5] undecyl-3-carbonyl) phenyl) dihydropyrimidine-2, 4(1H,3H)-dione (200 mg, 0.25 mmol), potassium carbonate (155 mg, 1.25 mmol) and potassium iodide (19 mg, 0.11 mmol) in acetone (4 mL). The reaction mixture was stirred at 80° C. for 3h and then concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 120 mg of target compound as a white solid.

[1448] LC-MS: (ESI, m/z): [M+H].sup.+=489.3

Step 2: Preparation of 2-(9-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-3, 9-diazaspiro [5.5] undec-3-yl) acetaldehyde

[1449] ##STR00446##

[1450] A mixture of 1-(5-(9-(2, 2-dimethoxyethyl)-3, 9-diazaspiro [5.5]undecyl-3-carbonyl)-2-methoxyphenyl) dihydropyrimidine-2, 4(1H,3H)-dione (120 mg, 0.246 mmol) in trifluoroacetic acid/dichloromethane (1 mL/2 mL) was stirred and reacted at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 100 mg of target compound as a white solid.

[1451] LC-MS: (ESI, m/z): [M+H].sup.+=443.1

[1452] The following intermediates were prepared with reference to intermediate 27:

TABLE-US-00005 Intermediate Structure Name LC-MS Intermediate 28 [00447] 3-(9-(3-(2,4- dioxotetrahydropyrimidin- 1(2H)-yl)- 4-methoxybenzoyl)- 3,9-diazaspiro[5.5] undec-3-yl) propionaldehyde (ES, m/z): [M + H].sup.+ = 457.3 Intermediate 29 [00448] 2-(7-(3-(2,4- dioxotetrahydropyrimidin- 1(2H)-yl)-4- methoxybenzoyl)- 2,7-diazaspiro[3.5] non-2-yl)acetaldehyde (ESI, m/z): [M + H].sup.+ = 415.1 Intermediate 30 [00449] 2-(2-(3-(2,4- dioxotetrahydropyrimidin- 1(2H)- yl)-4-methoxybenzoyl)- 2,7-diazaspiro[3.5]non- 7-yl)acetaldehyde (ESI, m/z): [M + H].sup.+ = 415.1 Intermediate 31 [00450] 2-(9-(4-chloro-3-(2,4- dioxotetrahydropyrimidin- 1(2H)-yl)benzoyl)-3,9- diazaspiro[5.5]undec-3-yl)ace- taldehyde (ES, m/z): [M + H].sup.+ = 447.1

Intermediate 32: 4-(9-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) butyraldehyde

Step 1: Preparation of 1-(5-(9-(3-(1, 3-dioxan-2-yl) propyl)-3, 9-diazaspiro[5.5]undecyl-3-carbonyl)-2-methoxyphenyl) dihydropyrimidine-2, 4(1H,3H)-dione

[1453] ##STR00451##

[1454] 2-(3-Bromopropyl)-1, 3-dioxane (58 mg, 0.3 mmol) was added to a mixture of 1-(2-methoxy-5-(3, 9-diazaspiro [5.5] undecyl-3-carbonyl) phenyl) dihydropyrimidine-2, 4(1H,3H)-dione (200 mg, 0.25 mmol), potassium carbonate (160 mg, 1.25 mmol) and potassium iodide (19 mg, 0.125 mmol) in acetone (10 mL). The reaction solution was stirred at 80° C. for 3h, and the resulting mixture was concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 70 mg of target compound as a white solid.

[1455] LC-MS: (ESI, m/z): [M+H].sup.+=515.1

Step 2: Preparation of 4-(9-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-3, 9-diazaspiro [5.5] undec-3-yl) butyraldehyde

[1456] ##STR00452##

[1457] A mixture of 1-(5-(9-(3-(1, 3-dioxacyclopentan-2-yl) propyl)-3, 9-diazaspiro[5.5]undecyl-3-carbonyl)-2-methoxyphenyl) dihydropyrimidine-2, 4(1H,3H)-dione (70 mg, 0.136 mmol) in trifluoroacetic acid/dichloromethane (1 mL/2 mL) was stirred for 4h at room temperature. The reaction solution was concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 60 mg of target compound as a white solid.

[1458] LC-MS: (ESI, m/z): [M+H].sup.+=471.1

Intermediate 33:3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxy-N-methyl-N-(5-oxopentyl) benzamide

Step 1: Preparation of tert-butyl (5-((tetrahydro-2H-pyran-2-yl) oxy) pentyl) carbamate

[1459] ##STR00453##

[1460] Tert-butyl (5-hydroxypentyl) carbamate (1.0 g, 4.92 mmol), dichloromethane (10.0 mL), p-toluenesulfonic acid (99.47 mg, 0.49 mmol) were added to a 50 mL round bottom flask, followed by 3, 4-dihydro-2H-pyran (496.0 mg, 5.90 mmol). The reaction mixture was stirred at room temperature overnight. 30 mL of water was added to quench the reaction, extracted with dichloromethane (2×30 mL), organic layers were combined, and washed with saturated saline (30 mL). The organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 900 mg of target product.

[1461] LC-MS: (ES, m/z): [M+H].sup.+=288.2

Step 2: Preparation of tert-butyl methyl (5-((tetrahydro-2H-pyran-2-yl) oxy) pentyl) carbamate

[1462] ##STR00454##

[1463] Tert-butyl (5-((tetrahydro-2H-pyran-2-yl) oxy) pentyl) carbamate (900.0 mg, 3.13 mmol), N,N-dimethylformamide (5.0 mL), and tetrahydrofuran (5.0 mL) were added to a 50 mL round-bottomed flask and placed in an ice-water bath, followed by sodium hydride (60% mineral oil mixture, 137.72 mg, 3.44 mmol). The reaction mixture was stirred at 0° C. for 0.5h. Potassium iodide (533.35 mg, 3.75 mmol) was then added and stirred overnight at room temperature. 30 mL of water was added to quench the reaction, extracted with ethyl acetate (2×30 mL), organic layers were combined, and washed with 30 mL of water and 30 mL of saturated saline. The organic layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 400 mg of target product.

[1464] LC-MS: [M+Na].sup.+=324.2

Step 3: Preparation of 5-(methylamino) pentyl-1-ol

[1465] ##STR00455##

[1466] Tert-butyl methyl (5-((tetrahydro-2H-pyran-2-yl) oxy) pentyl) carbamate (400 mg, 1.33 mmol), and dichloromethane (5.00 mL) were added to a 25 mL round bottom flask, followed by trifluoroacetic acid (3.0 mL), and the reaction mixture was stirred at room temperature for 2h. The reaction solution was concentrated under reduced pressure to obtain 500 mg of target product. The target product was directly used in the next reaction.

Step 4: Preparation of 3-(2,4-dioxotetrahydropyrimidin-1 (2H)-yl)-N-(5-hydroxypentyl)-4-methoxy-N-methylbenzamide

[1467] ##STR00456##

[1468] 5-(Methylamino) pentyl-1-ol (500 mg crude product, 1.33 mmol), dimethyl sulfoxide (5.0 mL), and N,N-diisopropylethylamine (686.28 mg, 5.32 mmol) were added to a 50 mL round bottom flask, followed by pentafluorophenyl 3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoate (430.0 mg, 1.0 mmol), the mixed reaction solution was stirred at room temperature overnight. 10 mL of water was added to quench the reaction. The mixture was extracted with ethyl acetate (2-20 mL), organic layers were combined, and washed with 10 mL of water and 20 mL of saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 100 mg of the target product.

Step 5: Preparation of 3-(2,4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxy-N-methyl-N-(5-oxopentyl) benzamide

[1469] ##STR00457##

[1470] 3-(2, 4-Dioxotetrahydropyrimidin-1(2H)-yl)-N-(5-hydroxypentyl)-4-methoxy-N-methylbenzamide (100 mg, 0.27 mmol) and dichloromethane (5.0 mL) were added to a 25 mL round-bottomed flask and placed in an ice-water bath, followed by the addition of a Dess-Martin oxidant (137.37 mg, 0.32 mmol), and the reaction mixture was stirred at room temperature for 2 hours. 5 mL of saturated sodium thiosulfate and 5 mL of saturated sodium bicarbonate were added to quench the reaction, and then the mixture was continued to be stirred for 20 minutes. The mixture was extracted with dichloromethane (2×20 mL), organic layers were combined and washed with 20 mL saturated saline. The organic layer was collected and dried over anhydrous sodium sulfate, then concentrated under reduced pressure to obtain 110.0 mg of the target crude product.

[1471] LC-MS: (ES, m/z): [M+H].sup.+=362.2

Intermediate 34: 2-(1-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl) piperidin-4-yl) acetaldehyde

Step 1: Preparation of 2-(piperidin-4-yl) acetaldehyde

[1472] ##STR00458##

[1473] Trifluoroacetic acid (1.5 mL) was added to a solution of tert-butyl 4-(2-oxoethyl) piperidin-1-carboxylate (580 mg, 2.72 mol) in dichloromethane (10 mL). The reaction solution was stirred at room temperature for 2 hours. The reactants were concentrated under reduced pressure to obtain 320 mg of oil, which was directly used in the next reaction.

[1474] LC-MS: (ES, m/z): [M+H].sup.+=128.2

Step 2: Preparation of 2-(1-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl) piperidin-4-yl) acetaldehyde

[1475] ##STR00459##

[1476] Triethylamine (406 mg, 3.98 mmol) and HATU (605 mg, 1.59 mmol) were added to a mixture of 2-(piperidin-4-yl) acetaldehyde (320 mg, 1.33 mmol) and 3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoic acid (150 mg, 1.33 mmol) in N,N-dimethylformamide (5 mL). The reaction solution was stirred at room temperature for 12 hours. The reaction solution was poured into water (50 mL), extracted with ethyl acetate (50 mL 2), and the organic layer was collected, washed with saline (50 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column to obtain 120 mg of target product as a yellow solid.

[1477] LC-MS: (ES, m/z): [M+H].sup.+=374.2

Intermediate 35:2-(4-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl) piperazin-1-yl) acetaldehyde

Step 1: Preparation of tert-butyl 4-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl) piperazine-1-carboxylate

[1478] ##STR00460##

[1479] N,N-diisopropylethylamine (440 mg, 9.3 mmol) and HATU (520 mg, 1.36 mmol) were added to a mixture of 3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoic acid (300 mg, 1.14 mmol) and tert-butyl piperazine-1-carboxylate (212 mg, 1.14 mmol) in N,N-dimethylformamide (10 mL). The reaction solution was stirred at room temperature for 2h, then the reaction solution was poured into water (20 mL) and extracted with ethyl acetate (20 mL×2). The organic layers were combined, washed with saline (20 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column to obtain 400 mg of target product as a yellow solid.

[1480] LC-MS: (ES, m/z): [M+H].sup.+=433.2

Step 2: Preparation of 1-(2-methoxy-5-(piperazin-1-carbonyl) phenyl) dihydropyrimidine-2,4(1H,3H)-dione

[1481] ##STR00461##

[1482] Trifluoroacetic acid (1.5 mL) was added to a solution of tert-butyl 4-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl) piperazine-1-carboxylate (400 mg, 0.93 mmol) in dichloromethane (10 mL). The reaction solution was stirred at room temperature for 2 hours. The solution was concentrated under reduced pressure to obtain 300 mg of crude target product as an oil. The crude product was used directly for the next step.

[1483] LC-MS: (ES, m/z): [M+H].sup.+=333.2

Step 3: Preparation of 1-(5-(4-(2, 2-dimethoxyethyl) piperazin-1-carbonyl)-2-methoxyphenyl) dihydropyrimidine-2,4 (1H,3H)-dione

[1484] ##STR00462##

[1485] Potassium carbonate (371 mg, 2.71 mmol) and potassium iodide (15 mg, 0.01 mmol) were added to a mixture of 1-(2-methoxy-5-(piperazin-1-carbonyl) phenyl) dihydropyrimidine-2,4(1H,3H)-dione (300 mg, 0.9 mmol) and 2-bromo-1, 1-dimethoxyethane (305 mg, 1.81 mmol) in acetone (20 mL). The reaction solution was stirred at 80° C. for 12h. The reaction solution was concentrated under reduced pressure to remove the solvent, and water (10 mL) and ethyl acetate (10 mL) were added. The water layers were separated and extracted with ethyl acetate (10 mL×2), and the organic layers were combined, washed with saline (10 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column to obtain 120 mg of target product as an oil.

[1486] LC-MS: (ES, m/z): [M+H].sup.+=421.2

Step 4: Preparation of 2-(4-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl) piperazin-1-yl) acetaldehyde

[1487] ##STR00463##

[1488] Trifluoroacetic acid (6 mL) was added to a solution of 1-(5-(4-(2, 2-dimethoxyethyl) piperazin-1-carbonyl)-2-methoxyphenyl) dihydropyrimidine-2,4 (1H,3H)-dione (120 mg, 0.27 mmol) in dichloromethane (10 mL). The reaction solution was stirred at room temperature for 12h. The solvent was removed by concentration under reduced pressure to obtain 53 mg of the crude target product. The crude product was used directly for the next step.

[1489] LC-MS: (ES, m/z): [M+H].sup.+=375.1

Intermediate 36: 3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxy-N-(2-(2-(2-oxoethoxy) ethoxy) ethyl) benzamide

Step 1: Preparation of 3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-N-(2-(2-(2-(2-hydroxyethoxy) ethoxy) ethyl)-4-methoxybenzamide

[1490] ##STR00464##

[1491] A mixture of 2-(2-(2-aminoethoxy) ethoxy) ethan-1-ol (200 mg, 0.75 mmol), 3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoic acid (135 mg, 0.91 mmol), HATU (432 mg, 1.14 mmol), and N,N-diisopropylethylamine (293 mg, 2.27 mmol) in N,N-dimethylformamide (5 mL) was stirred for 16h at room temperature. The reactants were poured into water (30 mL) and extracted with ethyl acetate (3×30 mL), the organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography to give 190 mg of product as yellow solid.

[1492] LC-MS-3: (ES, m/z): [M−H].sup.+=394.1

Step 2: Preparation of 3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxy-N-(2-(2-(2-oxoethoxy) ethoxy) ethyl) benzamide

[1493] ##STR00465##

[1494] Dess-Martin (408 mg, 0.96 mmol) was added to a mixture of 3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-N-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)-4-meth oxybenzamide (190 mg, 0.48 mmol) in dichloromethane (5 mL) at 0° C. The reaction solution was stirred for 2h at room temperature, filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 100 mg of target product as a yellow solid.

[1495] LC-MS-4: (ES, m/z): [M−H].sup.+=394.1

Intermediate 37: 5-((tert-butoxycarbonyl) amino) pentyl-4-methylbenzenesulfonate

[1496] ##STR00466##

[1497] A mixture of tert-butyl (5-hydroxypentyl) aminocarboxylate (500 mg, 2.463 mmol), p-toluenesulfonyl chloride (706 mg, 3.695 mmol) and 4-dimethylaminopyridine (154 mg, 1.232 mmol) in pyridine (5 mL) was stirred for 3 h at room temperature. The obtained reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (3-50 mL), and the organic phase was collected. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 400 mg of crude target product as a light yellow oil. The crude product was used directly for the next step.

[1498] LC-MS: (ESI, m/z): [M+H].sup.+=358.2

Intermediate 38: tert-butyl 9-(2-hydroxyethyl)-3-azaspiro[5.5] undecyl-3-carboxylate

Step 1: tert-butyl 9-(2-ethoxy-2-oxoethylene)-3-azaspiro[5.5] undecyl-3-carboxylate

[1499] ##STR00467##

[1500] At 0° C., sodium hydride (60% mineral oil mixture, 224 mg, 5.6 mmol) was added to a solution of 2-(diethoxyphosphoryl) ethyl acetate (1.26 g, 5.63 mmol) in N,N-dimethylformamide (15 mL), and stirred at 0° C. for 0.5 h, then tert-butyl 9-oxo-3-azaspiro [5.5]undecyl-3-carboxylate (1.0 g, 3.75 mmol) was added. The reaction mixture was stirred at 0° C. for 3h. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (3×30 mL). The organic layer was collected and washed with water (2×20 mL) and saturated saline (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain 0.7 g of the crude target product. The crude product was used directly for the next step.

Step 2: tert-butyl 9-(2-ethoxy-2-oxoethyl)-3-azaspiro[5.5] undecyl-3-carboxylate

[1501] ##STR00468##

[1502] The reaction mixture of tert-butyl 9-(2-ethoxy-2-oxoethylene)-3-azaspiro[5.5]undecyl-3-carboxylate (0.7 g, 2.08 mmol) and palladium hydroxide (0.2 g, 1.43 mmol) in ethanol (10 mL) was stirred at room temperature for 16h under hydrogen atmosphere. The reaction mixture was filtered and concentrated under reduced pressure to obtain 0.5 g of the crude target product. The crude product was used directly for the next step.

[1503] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 4.12 (q, J=7.1 Hz, 2H), 3.35 (ddd, J=7.6, 6.8, 5.0 Hz, 4H), 2.20 (d, J=7.1 Hz, 2H), 1.70-1.63 (m, 2H), 1.58 (dd, J=12.7, 7.1 Hz, 2H), 1.45 (s, 9H), 1.27 (dt, J=14.3, 6.5 Hz, 6H), 1.18-1.10 (m, 4H), 0.86 (dd, J=13.4, 6.3 Hz, 2H).

Step 3: tert-butyl 9-(2-hydroxyethyl)-3-azaspiro[5.5] undecyl-3-carboxylate

[1504] ##STR00469##

[1505] The reaction mixture of tert-butyl 9-(2-ethoxy-2-oxoethyl)-3-azaspiro[5.5]undecyl-3-carboxylate (0.5 g, 1.47 mmol) and lithium borohydride (0.13 g, 5.9 mmol) in tetrahydrofuran (5 mL) was refluxed overnight. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3×30 mL). The organic layer was collected and washed with water (2×20 mL) and saturated saline (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain 0.4 g of the crude target product.

[1506] LC-MS: (ESI, m/z): [M+H].sup.+=298.2

[1507] .sup.1H NMR (400 MHz, MeOD) δ 3.58 (t, J=6.6 Hz, 2H), 3.43-3.28 (m, 5H), 1.77-1.66 (m, 2H), 1.62-1.52 (m, 2H), 1.52-1.41 (m, 13H), 1.29 (t, J=5.6 Hz, 3H), 1.12 (dd, J=19.2, 11.5 Hz, 4H).

[1508] The following intermediates were prepared with reference to the method of intermediate 38:

TABLE-US-00006 Intermediate Structure Name LC-MS Intermediate 39 [00470] tert-butyl 2-(2-hydroxyethyl)-7-azaspiro[3.5] nonyl-7-carboxylate (ESI, m/z): [M + H].sup.+ = 270.3

Intermediate 40: N-(2-(1-(5-aminopentyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridine carboxamide

Step 1: Preparation of tert-butyl (5-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) pentyl) carbamate

[1509] ##STR00471##

[1510] A mixture of N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (387 mg, 0.924 mmol), 5-((tert-butoxycarbonyl) amino) pentyl-4-methylbenzenesulfonate (330 mg, 0.924 mmol) and potassium carbonate (638 mg, 4.622 mmol) in N,N-dimethylformamide (10 mL) was stirred at 75° C. for 3 hours, and the reaction solution was poured into water (100 mL), extracted with ethyl acetate (3-100 mL), the organic phase was collected, and washed with water (2×200 mL) and saline (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain yellow oily product (60 mg, Y=11%).

[1511] LC-MS: (ESI, m/z): [M+H].sup.+=605.3

Step 2: Preparation of N-(2-(1-(5-aminopentyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1512] ##STR00472##

[1513] A mixture of tert-butyl (5-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) pentyl) carbamate (60 mg, 0.0993 mmol) in trifluoroacetic acid/dichloromethane (1 mL, 1:1) was stirred and reacted overnight at room temperature, and the reactants were concentrated under reduced pressure to obtain 160 mg of crude target product as a yellow oil, and the crude product was directly used for the next step.

[1514] LC-MS: (ESI, m/z): [M+H].sup.+=505.2

Intermediate 41: N-(2-(1-(2-(2-aminoethoxy) ethyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: Preparation of tert-butyl (2-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) ethoxy) ethyl) carbamate

[1515] ##STR00473##

[1516] A solution of N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (500 mg, 1.193 mmol), tert-butyl (2-(2-bromoethoxy) ethyl) carbamate (610 mg, 2.864 mmol), potassium carbonate (823 mg, 5.967 mmol) and potassium iodide (99 mg, 0.597 mmol) in acetonitrile (10 mL) was stirred at 80° C. and reacted for 5 hours. The resulting reaction solution was diluted with ethyl acetate (50 mL), the organic phase was collected, washed with water (2×50 mL) and brine (50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 330 mg of yellow oily product.

[1517] LC-MS: (ESI, m/z): [M+H].sup.+=607.3

Step 2: Preparation of N-(2-(1-(2-(2-aminoethoxy) ethyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1518] ##STR00474##

[1519] A mixture of tert-butyl (2-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) ethoxy) ethyl) carbamate (140 mg, 0.231 mmol) in trifluoroacetic acid/dichloromethane (3 mL, 1:1) was stirred and reacted overnight at room temperature. The reactants were concentrated under reduced pressure to obtain 270 mg of crude product as a yellow oil. The crude product was used directly for the next step.

[1520] LC-MS: (ESI, m/z): [M+H].sup.+=507.3

Intermediate 42: N-(6-methoxy-2-(1-(2-(piperidin-4-yl) ethyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: Preparation of tert-butyl 4-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) ethyl) piperidine-1-formate

[1521] ##STR00475##

[1522] Sodium triacetoxyborohydride (1.518 g, 7.160 mmol) was added to a mixture of N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (1.0 g, 2.386 mmol) and tert-butyl 4-(2-oxoethyl) piperidine-1-carboxylate (0.813 g, 3.581 mmol) in tetrahydrofuran (20 mL). The reaction solution was stirred for reaction overnight at room temperature. The reaction solution was diluted with ethyl acetate (50 mL), washed with water (2×50 mL) and saturated saline (50 mL), and the organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 700 mg of target product as a yellow solid.

[1523] LC-MS: (ESI, m/z): [M+H].sup.+=631.3

Step 2: Preparation of N-(6-methoxy-2-(1-(2-(piperidin-4-yl) ethyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1524] ##STR00476##

[1525] A mixture of tert-butyl 4-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) ethyl) piperidine-1-formate (700 mg, 1.11 mmol) in hydrochloric acid/ethyl acetate (1M, 20 mL) was stirred and reacted overnight at room temperature. The reactants were concentrated under reduced pressure to obtain 650 mg of crude target product a yellow oil. The crude product was used directly for the next step.

[1526] LC-MS: (ESI, m/z): [M+H].sup.+=531.3

Intermediate 43: N-(6-methoxy-2-(1-(2-oxoethyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: Preparation of N-(2-(1-(2, 2-dimethoxyethyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1527] ##STR00477##

[1528] Potassium carbonate (1.235 g, 8.95 mmol) and potassium iodide (149 mg, 0.89 mmol) were added to a mixture of N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (750 mg, 1.79 mmol) and 2-bromo-1,1-dimethoxyethane (46 mg, 0.3 mmol) in acetonitrile (20 mL). The reaction solution was stirred at 80° C. for reaction overnight. The reaction solution was concentrated under reduced pressure to remove the solvent, water (20 mL) was added, extracted with ethyl acetate (20 mL×3), and the organic layers were combined, washed with saturated saline (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column to obtain 500 mg of target product as an oil.

[1529] LC-MS: (ESI, m/z): [M+H].sup.+=508.1

Step 2: Preparation of N-(6-methoxy-2-(1-(2-oxoethyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl)pyridinecarboxamide

[1530] ##STR00478##

[1531] A mixture of N-(2-(1-(2, 2-dimethoxyethyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (500 mg, 0.99 mmol) in hydrochloric acid (4 mL)/dioxane (5 mL) was stirred and reacted overnight at 50° C. The reaction solution was concentrated under reduced pressure to remove the solvent, water (20 mL) was added, extracted with ethyl acetate (20 mL-3), and the organic layers were combined, washed with saturated saline (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column to obtain 300 mg of target product as an oil.

[1532] LC-MS: (ESI, m/z): [M+H].sup.+=462.1

[1533] The following intermediates were prepared with reference to the method of intermediate 43:

TABLE-US-00007 Intermediate Structure Name LC-MS Intermediate 44 [00479] N-(6-(2-hydroxyprop- 2-yl)-2-(1-(2-oxoethyl) piperidin-4-yl)-2H-indazol-5-yl)- 6-(trifluoromethyl) pyridinecarboxamide (ES, m/z): [M + H].sup.+ = 490.2

Intermediate 45: N-(6-methoxy-2-(1-(3-piperidin-4-yl) propyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: tert-butyl 4-(3-(p-toluenesulfonyloxy) propyl) piperidine-1-formate

[1534] ##STR00480##

[1535] Triethylamine (2.1 g, 20.6 mmol) and p-toluenesulfonyl chloride (1.2 g, 6.1 mmol) were added to a stirred solution of tert-butyl 4-(3-hydroxypropyl) piperidine-1-formate (1.0 g, 4.1 mmol) in dichloromethane (30 mL). After the reaction mixture was stirred at 25° C. overnight, the reaction was quenched with saturated ammonium chloride (50 mL) and extracted with dichloromethane (2×50 mL). The organic layer was collected and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 1.1 g of target product.

[1536] LC-MS: (ESI, m/z): [M+H].sup.+=398.2.

Step 2: tert-butyl 4-(3-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido))-2H-indazol-2-yl) piperidin-1-yl) propyl) piperidine-1-formate

[1537] ##STR00481##

[1538] N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (1.2 g, 2.9 mmol) and potassium carbonate (2.0 g, 14.5 mmol) were added to a stirred solution of tert-butyl 4-(3-(p-toluenesulfonyloxy) propyl) piperidine-1-formate (1.1 g, 2.9 mmol) in N,N-dimethylformamide (15 mL). The reaction mixture was stirred at 75° C. for 3 hours. The reaction was quenched with water (100 mL) and extracted with ethyl acetate (3×50 mL). The organic layer was collected and washed with saturated saline (100 mL) and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 1.2 g of target product.

[1539] LC-MS: (ESI, m/z): [M+H].sup.+=645.3

Step 3: N-(6-methoxy-2-(1-(3-piperidin-4-yl) propyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1540] ##STR00482##

[1541] Trifluoroacetic acid (4.0 mL) was added to a stirred solution of tert-butyl 4-(3-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido))-2H-indazol-2-ye) piperidin-1-yl) propyl) piperidine-1-formate (1.4 g, 2.1 mmol) in dichloromethane (20.0 mL). The reaction mixture was stirred at 25° C. for 2 hours, then concentrated under reduced pressure to obtain 1.1 g of crude target product. The crude product was used directly for the next step.

[1542] LC-MS: (ESI, m/z): [M+H].sup.+=545.2.

[1543] The following intermediates were prepared with reference to the method of intermediate 45:

TABLE-US-00008 Inter- mediate Structure Name LC-MS Inter- mediate 46 [00483] N-(6-methoxy-2-(1-(3- piperazin-1-yl)propyl) piperidin-4-yl)-2H- indazol-5-yl)-6-(tri- fluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 546.3 Inter- mediate 47 [00484] N-(2-(1-(2-(3-aza- spiro[5.5]undec-9- yl)ethyl)piperidin-4-yl) 6-(2-hydroxyprop-2-yl)- 2H-indazol-5-yl)- 6-(trifluoromethyl) pyridinecarboxamide (ES, m/z): [M + H].sup.+ = 627.2 Inter- mediate 48 [00485] N-(2-(1-(2-(7-aza- spiro[3.5]non-2- yl)ethyl)piperidin- 4-yl)-6-(2-hydroxy- prop-2-yl)-2H-indazol-5- yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 599.3 Inter- mediate 49 [00486] N-(6-(2-hydroxyprop- 2-yl)-2-(1-(3-(piperidin-4- yl)propyl)piperidin- 4-yl)-2H-indazol- 5-yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 573.2 Inter- mediate 50 [00487] N-(6-(2-hydroxyprop- 2-yl)-2-(1-(2-(piperidin-4- yl)ethyl)piperidin- 4-yl)-2H-indazol- 5-yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 559.3 Inter- mediate 51 [00488] N-(2-(1-(2-(7-aza- spiro[3.5]non-2-yl)ethyl) piperidin-4-yl)-6-meth- oxy-2H-indazol-5-yl)-6- (trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+= 571.2 Inter- mediate 52 [00489] N-(2-(1-(2-(3-aza- spiro[5.5]undec-9- yl)ethyl]piperidin- 4-yl)-6-methoxy-2H- indazol-5-yl)-6-(trifluo- romethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+= 599.1

Intermediate 53: N-(6-methoxy-2-(1-(2-(piperidin-4-yloxy) ethyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: tert-butyl 4-(allyloxy) piperidine-1-carboxylate

[1544] ##STR00490##

[1545] Sodium hydride (60% mineral oil mixture, 360 mg, 12 mmol) was added to a stirred solution of tert-butyl 4-hydroxypiperidine-1-formate (2.0 g, 10 mmol) in tetrahydrofuran (50 mL) at 0° C., and then stirred for 1 h at 0° C. Then 3-bromoprop-1l-ene (1.44 g, 12 mmol) was added. After the stirring reaction was continued for 4h, the reaction was quenched with water (300 mL) and extracted with ethyl acetate (3×200 mL). The organic phase was collected and concentrated under reduced pressure. The concentrate was purified on a silica gel column to obtain 2.2 g of the target product.

[1546] .sup.1H NMR (400 MHz, CDCl3) δ 6.02-5.81 (m, 1H), 5.23 (dddd, J=42.1, 10.4, 3.1, 1.5 Hz, 2H), 4.02 (dt, J=5.5, 1.4 Hz, 2H), 3.86-3.71 (m, 2H), 3.49 (dt, J=12.0, 4.1 Hz, 1H), 3.15-2.97 (m, 2H), 1.91-1.74 (m, 2H), 1.60-1.36 (m, 11H).

Step 2: tert-butyl 4-(2-oxoethoxy) piperidine-1-carboxylate

[1547] ##STR00491##

[1548] Under ozone atmosphere, tert-butyl 4-(allyloxy) piperidine-1-carboxylate (1 g, 4.15 mmol) in dichloromethane (30 mL) was reacted at −78° C. for 30 minutes and then quenched with dimethyl sulfide (10 mL), concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 300 mg of the target product.

[1549] .sup.1H NMR (400 MHz, CDCl3) δ 9.74 (t, J=0.8 Hz, 1H), 4.68 (m, 1H), 4.51-4.43 (m, 1H), 3.56-3.52 (m, 2H), 3.29 (m, 2H), 3.21 (m, 1H), 2.31-2.19 (m, 2H), 1.82-1.68 (m, 2H), 1.46 (s, 9H).

[1550] Step 3: tert-butyl 4-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido))-2H-indazol-2-yl) piperidin-1-yl) ethoxy) piperidine-1-carboxylate

##STR00492##

[1551] Sodium triacetoxyborohydride (303 mg, 1.40 mmol) and three drops of acetic acid were added to a stirred solution of N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (200 mg, 0.477 mmol) and tert-butyl 4-(2-oxoethoxy) piperidine-1-carboxylate (174 mg, 0.716 mmol) in tetrahydrofuran (15 mL), then the reaction mixture was stirred at 25° C. for 2 hours. The reaction mixture was diluted with ethyl acetate (60 mL) and washed with water (2×60 mL) and saturated saline (60 mL). The organic phase was collected and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The concentrate was purified on a silica gel column to obtain 260 mg of the target product.

[1552] LC-MS: (ESI, m/z): [M+H].sup.+=647.1.

Step 4: N-(6-methoxy-2-(1-(2-(piperidin-4-yloxy) ethyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1553] ##STR00493##

[1554] The reaction mixture of tert-butyl 4-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido))-2H-indazol-2-yl) piperidin-1-yl) ethoxy) piperidine-1-formate (260 mg, 0.402 mmol) in trifluoroacetic acid/dichloromethane (4 mL, 1:1) was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to obtain 500 mg of the crude target product. The crude product was used directly for the next step.

[1555] LC-MS: (ESI, m/z): [M+H].sup.+=547.3.

[1556] The following intermediates were prepared with reference to the method of intermediate 53:

TABLE-US-00009 Inter- mediate Structure Name LC-MS Inter- mediate 54 [00494] N-(6-(2-hydroxyprop-2-yl)-2-(1- (2-(piperidin-4-yloxy)ethyl) piperidin-4-yl)-2H-indazol-5- yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 575.2

Intermediate 55: N-(2-(1-((3-azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: tert-butyl 9-((4-(6-(2-hydroxyprop-2-yl)-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5]undecyl-3-carboxylate

[1557] ##STR00495##

[1558] At room temperature, sodium triacetylborohydride (212 mg, 1 mmol) was added to a mixture of N-(6-(2-hydroxyprop-2-yl)-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (223 mg, 0.5 mmol) and tert-butyl 9-formyl-3-azaspiro[5.5]undecyl-3-carboxylate (140 mg, 0.5 mmol) in tetrahydrofuran (5 mL), and then stirred and reacted for 2 hours. The solution was diluted with water (20 mL) and extracted with dichloromethane (10 mL×3). The organic layer was collected, concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 260 mg of the target product.

[1559] LC-MS: (ESI, m/z): [M+H].sup.+=713.1.

[1560] .sup.1H NMR (400 MHz, DMSO) δ 12.37 (s, 1H), 8.72 (s, 1H), 8.45 (d, J=7.7 Hz, 1H), 8.37 (dd, J=14.8, 7.0 Hz, 2H), 8.16 (dd, J=7.8, 0.8 Hz, 1H), 7.57 (s, 1H), 5.95 (s, 1H), 4.44 (s, 1H), 3.28 (s, 4H), 2.96 (s, 2H), 2.08 (d, J=10.9 Hz, 7H), 1.70-1.44 (m, 12H), 1.39 (s, 11H), 1.22 (d, J=6.0 Hz, 2H), 1.07 (d, J=9.7 Hz, 4H).

Step 2: N-(2-(1-((3-azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1561] ##STR00496##

[1562] Tert-butyl 9-((4-(6-(2-hydroxyprop-2-yl)-5-(6-(trifluromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5, 5] undecyl-3-carboxylate (260 mg, 0.365 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (1 mL) was added at 0° C. The reaction mixture was stirred and reacted at 0° C. for 2h. N,N-diisopropylethylamine was slowly added at 0-5° C. until the pH of the reaction solution was >9, the solution was concentrated under reduced pressure at room temperature, and the crude product was directly used for the next step without further purification.

[1563] LC-MS: (ESL m/z): [M+H].sup.+=613.1.

[1564] The following intermediates were prepared with reference to the method of intermediate 55:

TABLE-US-00010 Intermediate Structure Name LC-MS Intermediate 56 [00497] N-(2-(1-(((7-azaspiro[3.5] non-2-yl) methyl) piperidin-4-yl)-6-(2-hy- droxyprop-2-yl)-2H-indazol- 5-yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H] .sup.+ = 585.3 Intermediate 57 [00498] N-(2-(1-(((2-azaspiro[3.5] non-7-yl) methyl) piperidin-4-yl)-6-methoxy- 2H-indazol-5-yl)-6-(tri- fluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H] .sup.+ = 557.3 Intermediate 58 [00499] N-(2-(1-((3-azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6-methoxy- 2H-indazol-5-yl)-6-(trifluoro- methyl) pyridinecarboxamide (ESI, m/z): [M + H] .sup.+ = 585.3 Intermediate 59 [00500] N-(2-(1-((3-azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6-(trifluoro- methoxy)-2H-indazol-5- yl)-6-(trifluoromethyl)) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 639.2

Intermediate 60: N-(6-methoxy-2-(1-(2-(piperazin-1-yl) ethyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: Preparation of tert-butyl 4-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido))-2H-indazol-2-yl) piperidin-1-yl) ethyl) piperazine-1-carboxylate

[1565] ##STR00501##

[1566] Sodium triacetoxyborohydride (110 mg, 0.521 mmol) was added to a mixture of N-(6-methoxy-2-(1-(2-oxoethyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (80 mg, 0.174 mmol) and tert-butyl piperazine-1-formate (32 mg, 0.174 mmol) in tetrahydrofuran (2 mL). The reaction solution was stirred and reacted overnight at room temperature, the resulting reaction solution was diluted with ethyl acetate (20 mL), washed with water (2-20 mL) and saturated saline (20 mL), and the organic phase was collected. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 80 mg of target product as a yellow oil.

[1567] LC-MS: (ESI, m/z): [M+H].sup.+=632.1

Step 2: N-(6-methoxy-2-(1-(2-(piperazin-1-yl) ethyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1568] ##STR00502##

[1569] A mixture of tert-butyl 4-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido))-2H-indazol-2-yl) piperidin-1-yl) ethyl) piperazine-1-carboxylate (80 mg, 0.127 mmol) in trifluoroacetic acid/dichloromethane (2 ml, 1:1) was stirred and reacted overnight at room temperature. The reaction solution was concentrated under reduced pressure to obtain 120 mg of crude target product as a yellow oil. The crude product was used directly for the next step.

[1570] LC-MS: (ESI, m/z): [M+H].sup.+=532.3

[1571] The following intermediates were prepared with reference to the method of intermediate 60:

TABLE-US-00011 Intermediate Structure Name LC-MS Intermediate 61 [00503] N-(2-(1-(2-(2,7-diazaspiro [3.5] non-2-yl) ethyl) piperidin-4-yl)-6-methoxy- 2H-indazol-5-yl)-6- (trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 572.3. Intermediate 62 [00504] N-(2-(1-(2-(3, 9-diazaspiro[5.5] undec-3-yl) ethyl) piperidin-4-yl)-6-methoxy- 2H-indazol-5-yl)-6- (trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 600.3 Intermediate 63 [00505] N-(2-(1-(2-(3,9-diazaspiro [5.5] undec-3-yl) ethyl) piperidin-4-yl)-6-(2-hydroxy- prop-2-yl)-2H-indazol-5- yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 628.3 Intermediate 64 [00506] N-(2-((1r,4r)-4-((3, 9-diazaspiro [5.5] undec-3-yl) methyl) cyclohexyl)-6-methoxy-2H- indazol-5-yl)-6- (trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 585.6. Intermediate 65 [00507] N-(2-((1r,4r)-4-(2-(3, 9-diazaspiro [5.5] undec-3-yl) ethyl) cyclohexyl)-6-methoxy-2H- indazol-5-yl)- 6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 599.3 Intermediate 67 [00508] N-(6-methoxy-2-((1r,4r)-4- (piperazin-1-ylmethyl) cyclohexyl)-2H-indazol-5- yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 517.2. Intermediate 68 [00509] N-(6-methoxy-  ((1r,4r)-4-(2-(piperazin- 1-yl) ethyl) cyclohexyl)-2H-indazol-5- yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 531.3 indicates data missing or illegible when filed

Intermediate 69: N-(6-methoxy-2-(1-(2-piperidin-4-yl) methyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: Preparation of tert-butyl 4-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) methyl) piperidine-1-formate

[1572] ##STR00510##

[1573] Sodium triacetoxyborohydride (1.518 g, 7.160 mmol) was added to a mixture of N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (1.0 g, 2.386 mmol) and tert-butyl 4-formylpiperidine-1-carboxylate (0.763 g, 3.581 mmol) in tetrahydrofuran (20 mL). The reaction solution was stirred for reaction overnight at room temperature. The reaction solution was diluted with ethyl acetate (50 mL), washed with water (2×50 mL) and saturated saline (50 mL), the organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 550 mg of target product as a yellow solid.

[1574] LC-MS: (ESI, m/z): [M+H].sup.+=617.3

Step 2: Preparation of N-(6-methoxy-2-(1-(2-(piperidin-4-yl) methyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1575] ##STR00511##

[1576] A mixture of tert-butyl 4-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) methyl) piperidine-1-formate (573 mg, 1.11 mmol) in hydrochloric acid/ethyl acetate (1M, 20 mL) was stirred and reacted overnight at room temperature. The reactants were concentrated under reduced pressure to obtain 400 mg of crude target product as a yellow oil. The crude product was used directly for the next step.

[1577] LC-MS: (ESI, m/z): [M+H].sup.+=517.2

Intermediate 70: N-(6-methoxy-2-(1-(2-(piperidin-4-yl) methyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: Preparation of tert-butyl 4-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) methyl) piperidine-1-formate

[1578] ##STR00512##

[1579] Sodium triacetoxyborohydride (1.518 g, 7.160 mmol) was added to a mixture of N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (1.0 g, 2.386 mmol) and tert-butyl 4-formylpiperidine-1-carboxylate (0.763 g, 3.581 mmol) in tetrahydrofuran (20 mL). The reaction solution was stirred for reaction overnight at room temperature. The reaction solution was diluted with ethyl acetate (50 mL), washed with water (2×50 mL) and saturated saline (50 mL), and the organic phase was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 550 mg of target product as a yellow solid.

[1580] LC-MS: (ESI, m/z): [M+H].sup.+=617.3

Step 2: Preparation of N-(6-methoxy-2-(1-(2-(piperidin-4-yl) methyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1581] ##STR00513##

[1582] A mixture of tert-butyl 4-(2-(4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) methyl) piperidine-1-formate (573 mg, 1.11 mmol) in hydrochloric acid/ethyl acetate (1M, 20 mL) was stirred and reacted overnight at room temperature. The reactants were concentrated under reduced pressure to obtain 400 mg of crude target product as a yellow oil. The crude product was used directly for the next step.

[1583] LC-MS: (ESI, m/z): [M+H].sup.+=517.2

Intermediate 71: N-(2-((1R,4R)-4-formylcyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridine carboxamide

Step 1: Preparation of ((1R,4R)-4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl) methanol

[1584] ##STR00514##

[1585] A mixture of 2-azido-4-methoxy-5-nitrobenzaldehyde (1.33 g, 6 mmol) and ((1r,4r)-4-aminocyclohexyl) methanol (774 mg, 6 mmol) in toluene (50 mL) was heated to 100° C. and stirred for reaction overnight. The reaction mixture was concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 1.5 g of target product as a brown solid.

[1586] .sup.1H NMR (400 MHz, DMSO) δ 8.60 (s, 1H), 8.39 (s, 1H), 7.27 (s, 1H), 4.54-4.40 (m, 2H), 3.91 (s, 3H), 3.29 (t, J=5.8 Hz, 2H), 2.20-2.09 (m, 2H), 1.96-1.82 (m, 4H), 1.47 (dtd, J=11.9, 6.0, 3.0 Hz, 1H), 1.22-1.04 (m, 2H).

Step 2: ((1R,4R)-4-(5-amino-6-methoxy-2H-indazol-2-yl) cyclohexyl) methanol

[1587] ##STR00515##

[1588] Hydrazine hydrate (2 mL) and Raney-Ni (150 mg) were added to a solution of ((1R,4R)-4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl) methanol (1.5 g, 5.0 mmol) in ethanol (50 mL). The reaction mixture was stirred at room temperature for 2 hours. Filtered, solid was washed with ethanol (10 mL/1). The filtrate was collected and concentrated under reduced pressure to obtain 1.4 g of crude product as a brown gel.

[1589] LC-MS: (ESI, m/z): [M+H].sup.+=276.3.

Step 3: N-(2-((1R,4R)-4-(hydroxymethyl) cyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1590] ##STR00516##

[1591] 6-(Trifluoromethyl) picolinic acid (0.97 g, 5.07 mmol), N,N-diisopropylethylamine (1.96 g, 15.21 mmol) and HATU (2.5 g, 6.59 mmol) were added to a solution of ((1R,4R)-4-(5-amino-6-methoxy-2H-indazol-2-yl) cyclohexyl) methanol (1.4 g, 5.07 mmol) in N,N-dimethylformamide (20 mL). After stirring the reaction mixture at room temperature for 2h, the reaction was quenched with water and extracted with ethyl acetate to collect the organic layer. The organic layer was washed with saturated saline, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 1.2 g of target product as a brown solid.

[1592] LC-MS: (ESI, m/z): [M+H].sup.+=449.2.

Step 4: N-(2-((1R,4R)-4-formylcyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1593] ##STR00517##

[1594] 2-Iodoacyl benzoic acid (404 mg, 1.0 mmol) was added to a mixture of N-(2-((1R,4R)-4-(hydroxymethyl) cyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (224 mg, 0.5 mmol) in acetonitrile (10 mL), the reaction mixture was heated to 80° C. and stirred for 2h. The reaction solution was concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 180 mg of target product as a brown solid.

[1595] LC-MS: (ESI, m/z): [M+H].sup.+=447.1

[1596] The following intermediates were prepared with reference to the method of intermediate 71:

TABLE-US-00012 Intermediate Structure Name LC-MS Intermediate 72 [00518] N-(2-((1r,4r)-4-for- mylcyclohexyl)-6- methoxy-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarboxamide LC-MS: (ESI, m/z): [M + H].sup.+ = 447.1. .sup.1H NMR (400 MHz, DMSO) δ 10.50 (s, 1H), 9.65 (s, 1H), 8.69 (s, 1H), 8.47 (d, J = 7.5 Hz, 1H), 8.41 (t, J = 7.8 Hz, 1H), 8.34 (s, 1H), 8.22 (dd, J = 7.6, 0.8 Hz, 1H), 7.16 (s, 1H), 4.40 (ddd, J = 11.7, 8.0, 3.9 Hz, 1H), 3.98 (s, 3H), 2.43 (t, J = 12.3 Hz, 1H), 2.16 (dd, J = 37.9, 10.4 Hz, 4H), 1.99-1.90 (m, 2H), 1.46 (tt, J = 13.1, 6.6 Hz, 2H). Intermediate 73 [00519] N-(6-methoxy-2-((1r, 4r)-4-(2-oxoethyl) cyclohexyl)-2H-inda- zol-5-yl)-6-(trifluoro- methyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 461.2

Intermediates 74: N-(2-((1r,4r)-4-formylcyclohexanyl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: ethyl (1s, 4s)-4-hydroxycyclohexan-1-carboxylate

[1597] ##STR00520##

[1598] Sulfuric acid (0.25 mL) was added to a solution of (1s, 4s)-4-hydroxycyclohexan-1-carboxylic acid (10 g, 69.4 mmol) in ethanol (50 mL), and the reaction mixture was stirred at 70° C. for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain 11.6 g of crude target product. The crude product was used directly for the next step.

Step 2: ethyl (1s, 4s)-4-(toluenesulfonyloxy) cyclohexane-1-carboxylate

[1599] ##STR00521##

[1600] p-Toluenesulfonyl chloride (12 g, 62 mmol), TEA (16 g, 156 mmol) and 4-dimethylaminopyridine (61 mg, 0.5 mmol) were added to a mixture of ethyl (1s, 4s)-4-hydroxycyclohexane-1-carboxylate (9.0 g, 12 mmol) in dichloromethane (150 mL). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 11 g of the target compound as a yellow solid.

[1601] LC-MS: (ES, m/z): [M+NH.sub.4].sup.+=344.2.

Step 3: (1s, 4s)-4-(hydroxymethyl) cyclohexyl 4-methylbenzenesulfonate

[1602] ##STR00522##

[1603] At 0° C., lithium aluminum hydride (24 mL) was added to a solution of ethyl (1s, 4s)-4-(toluenesulfonyloxy) cyclohexane-1-carboxylate (5.0 g, 15.3 mmol) in tetrahydrofuran (50 mL). The reaction mixture was stirred at 0° C. for 2 hours. Sodium sulfate decahydrate was added, filtered and the filtrate was concentrated to obtain 3.6 g of target compound as a colorless solid.

[1604] LC-MS: (ES, m/z): [M+HN.sub.4].sup.+=302.1.

Step 4: N-(2-((1r,4r)-4-(hydroxymethyl) cyclohexyl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1605] ##STR00523##

[1606] (1s, 4s)-4-(hydroxymethyl) cyclohexyl-4-methylbenzenesulfonate (2.65 g, 9 mmol) and cesium carbonate (8.8 g, 27 mmol) were added to a mixture of N-(6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl)pyridinecarboxamide (3.4 g, 9 mmol) in N,N-dimethylformamide (50 mL). The reaction mixture was stirred at 80° C. for 16 hours. The reaction mixture was quenched with water (100 mL) and the solution was extracted with ethyl acetate (3×30 mL). The organic layer was collected, washed with water (2×20 mL) and saturated saline (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 500 mg of target compound as a yellow solid.

[1607] LC-MS: (ES, m/z): [M+H].sup.+=477.1

Step 5: N-(2-((1r,4r)-4-formylcyclohexanyl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1608] ##STR00524##

[1609] Dess-Martin (213 mg, 0.50 mmol) was added to a mixture of N-(2-(1r,4r)-4-(hydroxymethyl) cyclohexyl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (200 mg, 0.42 mmol) in 5 mL of dichloromethane. After the reaction mixture was stirred at room temperature for 16 hours, the reaction mixture was quenched with water (10 mL) and extracted with dichloromethane (3×10 mL). The organic layer was collected, washed with water (2×2 mL) and saturated saline (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 50 mg of target compound as a yellow solid.

[1610] LC-MS: (ES, m/z): [M+H].sup.+=475.0.

[1611] Intermediate 75: N-(6-methoxy-2-((1R,4R)-4-((methyl (2-(piperidin-4-yl) ethyl) amino) methyl) cyclohexyl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: tert-butyl 4-(2-((((1R,4R)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) methyl) amino) ethyl) piperidine-1-formate

[1612] ##STR00525##

[1613] Sodium triacetoxyborohydride (424 mg, 2 mmol) was added to a mixture f N-(2-((1R,4R)-4-formylcyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (180 mg, 0.4 mmol) and butyl 4-(2-aminoethyl) piperidine-1-formate (91 mg, 0.4 mmol) in tetrahydrofuran (10 mL), and then the reaction mixture was stirred overnight at room temperature. Water (20 mL) was added, extracted with ethyl acetate (20 mL×3), the organic layer was collected, washed with saturated saline (20 mL×1), and the organic layer was collected, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was purified by a chromatographic column to obtain 170 mg of target product as a white solid.

[1614] LC-MS: (ESI, m/z): [M+H].sup.+=659.4.

Step 2: tert-butyl 4-(2-((((1R,4R)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridineformyl)-2H-indazol-2-yl) cyclohexyl) methyl) (methyl) amino) ethyl) piperidine-1-carboxylate

[1615] ##STR00526##

[1616] At room temperature, sodium triacetoxyborohydride (276 mg, 1.3 mmol) was added to a mixture of tert-butyl 4-(2-((((1R,4R)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) methyl) amino) ethyl) piperidine-1-formate (170 mg, 0.26 mmol) and formaldehyde (37 wt % aqueous solution, 162 mg, 2 mmol) in tetrahydrofuran (10 mL), and the reaction mixture was stirred for reaction overnight. Water (20 mL) was added to the reaction mixture, extracted with ethyl acetate (20 mL×3), the organic layer was collected, washed with saturated saline (20 mL×1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 135 mg of target product as a white solid.

[1617] LC-MS: (ESI, m/z): [M+H].sup.+=673.4.

Step 3: N-(6-methoxy-2-((1R,4R)-4-((methyl (2-(piperidin-4-yl) ethyl) amino) methyl) cyclohexyl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1618] ##STR00527##

[1619] At room temperature, trifluoroacetic acid (4 mL) was added to a mixture of tert-butyl 4-(2-((((1R,4R)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridineformyl)-2H-indazol-2-yl) cyclohexyl) methyl) (methyl) amino) ethyl) piperidine-1-carboxylate (135 mg, 0.20 mmol) in dichloromethane (10 mL), and the mixture was stirred for 2h. The reaction mixture was concentrated under reduced pressure to obtain 130 mg of crude product as a brown gel. The crude product can be used directly for the next step without further purification.

[1620] LC-MS: (ESI, m/z): [M+H].sup.+=573.4.

[1621] The following intermediates were prepared with reference to the method of intermediate 75:

TABLE-US-00013 Intermediate Structure Name LC-MS Intermediate 76 [00528] N-(6-methoxy-2-((1r,4r)- 4-((methyl (2-(piperidin-4-yl) ethyl) amino) methyl) cyclohexyl)-2H-indazol- 5-yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 573.3 Intermediate 77 [00529] N-(6-methoxy- 2-((1r,4r)-4-((methyl (3-azaspiro [5.5] undec-9-yl) amino) methyl) cyclohexyl)-2H-indazol- 5-yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 613.3 Intermediate 78 [00530] N-(6-(2-hydroxyprop- 2-yl)-2-((1r,4r)-4-((meth- yl (2-(piperidin-4-yl) ethyl) amino) methyl) cyclohexyl)-2H-indazol- 5-yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 601.3. Intermediate 79 [00531] N-(6-(2-hydroxyprop-2-yl)- 2-((1r,4r)-4-((methyl (piperidin-4-ylmethyl) amino) methyl) cyclohexyl)-2H-indazol- 5-yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 587.6. Intermediate 80 [00532] N-(6-(2-hydroxyprop-2-yl)- 2-((1r,4r)-4-((methyl (3-azaspiro [5.5] undec-9-yl) amino) methyl) cyclohexyl)-2H-indazol- 5-yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 641.5. Intermediate 81 [00533] N-(6-methoxy- 2-((1r,4r)-4-((methyl (piperidin-4-yl) amino) methyl) cyclohexyl)-2H-indazol- 5-yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 545.2 Intermediate 82 [00534] N-(6-methoxy-2-((1r,4r)- 4-((methyl (4-(piperidin-4-yl) butyl) amino) methyl) cyclohexyl)-2H-indazol- 5-yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 601.2.

Intermediate 83: N-(6-methoxy-2-((1r,4r)-4-(methyl (piperidin-4-ylmethyl amino) methyl) cyclohexyl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: tert-butyl 4-(((((1r,4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) methyl) (methyl) amino) methyl) piperidine-1-formate

[1622] ##STR00535##

[1623] A mixture of N-(2-((1r,4r)-4-formyl cyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (100 mg, 0.22 mmol), tert-butyl 4-((methylamino) methyl) piperidine-1-formate (51 mg, 0.22 mmol) and sodium triaceoxyborohydride (142 mg, 0.67 mmol) in tetrahydrofuran (10 mL) was stirred overnight at room temperature. The reaction mixture was quenched with water (50 mL) and extracted with dichloromethane (3×20 mL), the organic layer was collected, washed with saturated saline (60 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 120 mg of target product as a light yellow solid.

[1624] LC-MS: (ESI, m/z): [M+H].sup.+=659.4.

Step 2: N-(6-methoxy-2-((1r,4r)-4-((methyl (piperidin-4-ylmethyl) amino) methyl) cyclohexyl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1625] ##STR00536##

[1626] Trifluoroacetic acid (2 mL) was added to a mixture of tert-butyl 4-((((1r,4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) methyl) (methyl) amino) methyl) piperidine-1-formate (120 mg, 0.18 mmol) in dichloromethane (5 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated, to obtain 200 mg of crude target product as a light yellow solid was obtained. The crude product was used directly for the next step.

[1627] LC-MS: (ESI, m/z): [M+H].sup.+=559.2.

Intermediate 84: tert-butyl 9-amino-3-azaspiro[5.5] undecyl-3-carboxylate

Step 1: tert-butyl 9-(benzylamino)-3-azaspiro[5.5] undecyl-3-carboxylate

[1628] ##STR00537##

[1629] A mixture of tert-butyl 9-oxo-3-azaspiro[5.5] undecyl-3-carboxylate (267 mg, 1.0 mmol), benzylamine (321 mg, 3.0 mmol) and sodium triacetoxyborohydride (2.10 g, 10.0 mmol) in tetrahydrofuran (30 mL) was stirred for 2 hours at room temperature. The reaction mixture was quenched with water (100 mL) and extracted with dichloromethane (3×50 mL). The organic layer was collected and washed with saturated saline (150 mL), dried over anhydrous sodium sulfate, filtered and concentrated, and the concentrate was purified by column chromatography to obtain 310 mg of target compound as a colorless oil.

[1630] LC-MS: (ESI, m/z): [M+H].sup.+=359.2

Step 2: tert-butyl 9-amino-3-azaspiro[5.5] undecyl-3-carboxylate

[1631] ##STR00538##

[1632] Palladium hydroxide/carbon (303 mg) was added to a mixture of tert-butyl 9-(benzylamino)-3-azaspiro[5.5] undecyl-3-carboxylate (310 mg, 0.86 mmol) in methanol (30 mL). The reaction mixture was stirred overnight at room temperature in a hydrogen atmosphere. The reaction mixture was filtered through a diatomite mat and the filtrate was concentrated to obtain 200 mg of target crude product as a colorless oil. The crude mixture was used directly for the next step.

[1633] LC-MS: (ESI, m/z): [M+H].sup.+=269.3.

[1634] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 3.49 (s, 1H), 3.35 (d, J=3.3 Hz, 4H), 3.15 (s, 1H), 1.97 (d, J=12.3 Hz, 2H), 1.83-1.65 (m, 5H), 1.45 (s, 9H), 1.36-1.12 (m, 6H).

Intermediate 85: N-(6-methoxy-2-((1r,4r)-4-(methyl (3-azaspiro[5.5] undec-9-yl) amino) cyclohexyl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: tert-butyl ((1r,4r)-4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl) carbamate

[1635] ##STR00539##

[1636] Tert-butyl ((1r,4r)-4-aminocyclohexyl) carbamate (1.03 g, 4.8 mmol) was added to a solution of 2-azido-4-methoxy-5-nitrobenzaldehyde (1.07 g, 4.8 mmol) in toluene (50 mL), and the mixture was heated to 110° C. and stirred overnight. The reaction mixture was concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 1.5 g of target product as a yellow solid.

[1637] LC-MS: (ESI, m/z): [M+H].sup.+=391.2.

Step 2: tert-butyl ((1r,4r)-4-(5-amino-6-methoxy-2H-indazol-2-yl) cyclohexyl) carbamate

[1638] ##STR00540##

[1639] Raney-Ni (100 mg) and hydrazine hydrate (85%, 1.5 mL) were added to a mixture of tert-butyl ((1r,4r)-4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl) carbamate (1.5 g, 3.85 mmol) in ethanol (20 mL). The reaction mixture was stirred at room temperature for 2 hours, filtered, and the filtrate was concentrated under reduced pressure to obtain 1.15 g of the crude target product as a yellow gel, which can be used for the next step without further purification.

[1640] LC-MS: (ESI, m/z): [M+H].sup.+=361.2.

Step 3: tert-butyl ((1r, 4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) carbamate

[1641] ##STR00541##

[1642] 6-(Trifluoromethyl) picolinic acid (609 mg, 3.19 mmol), HATU (1.5 g, 4.0 mmol) and N,N-diisopropylethylamine (1.2 g, 9.3 mmol) were added to a mixture of tert-butyl ((1r,4r)-4-(5-amino-6-methoxy-2H-indazol-2-yl) cyclohexyl) carbamate (1.15 g, 3.19 mmol) in N,N-dimethylformamide (10 mL). The reaction mixture was stirred at room temperature for 2 hours, and the reaction was quenched with water (100 mL), and extracted with ethyl acetate (3×50 mL). The organic layers were collected and washed with saturated saline (150 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 1.3 g of crude target product as a yellow solid.

[1643] LC-MS: (ESI, m/z): [M+H].sup.+=534.3.

Step 4: N-(2-((1r,4r)-4-aminocyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1644] ##STR00542##

[1645] A mixture of tert-butyl ((1r, 4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) carbamate (400 mg, 0.75 mmol) in trifluoroacetic acid/dichloromethane (8 mL, 1:3) was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain 430 mg of the crude target product as a brown oil. The crude product was used directly for the next reaction.

[1646] LC-MS: (ESI, m/z): [M+H].sup.+=434.2.

Step 5: tert-butyl 9-(((1r,4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) amino)-3-azaspiro[5.5]undecyl-3-carboxylate

[1647] ##STR00543##

[1648] A mixture of N-(2-((1r,4r)-4-aminocyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (430 mg, crude), tert-butyl 9-oxo-3-azaspiro[5.5] undecyl-3-carboxylate (210 mg, 0.79 mmol) and sodium triacetoxyborohydride (835 mg, 3.93 mmol) in tetrahydrofuran (10 mL) was stirred overnight at room temperature. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (3×30 mL). The organic layer was collected, washed with saturated saline (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 400 mg of target product as a light yellow solid.

[1649] LC-MS: (ESI, m/z): [M+H].sup.+=685.4.

Step 6: tert-butyl 9-(((1r,4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) (methyl) amino)-3-azaspiro[5.5]undecyl-3-carboxylate

[1650] ##STR00544##

[1651] A mixture of tert-butyl 9-(((1r,4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) amino)-3-azaspiro[5.5]undecyl-3-carboxylate (400 mg, 0.58 mmol), formaldehyde (0.5 mL) and sodium triacetoxylborohydride (615 mg, 2.9 mmol) in tetrahydrofuran (10 mL) was stirred overnight at room temperature. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (3-30 mL). The organic layer was collected, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 300 mg of the target product as a yellow oil.

[1652] LC-MS: (ESI, m/z): [M+H].sup.+=699.3.

Step 7: N-(6-methoxy-2-((1r,4r)-4-(methyl (3-azaspiro[5.5] undec-9-yl) amino) cyclohexyl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1653] ##STR00545##

[1654] A mixture of tert-butyl 9-(((1r,4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) (methyl) amino)-3-azaspiro[5.5]undecyl-3-carboxylate (300 mg, 0.43 mmol) in trifluoroacetic acid/dichloromethane (6 mL, 1:2) was stirred and reacted at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain 330 mg of the crude target product as a brown oil. The crude product was used directly for the next step.

[1655] LC-MS: (ESI, m/z): [M+H].sup.+=599.4.

[1656] The following intermediates were prepared with reference to intermediate 85.

TABLE-US-00014 Intermediate Structure Name LC-MS Intermediate 86 [00546] N-(2-((1r,4r)-4-(ethyl (3-azaspiro[5.5] undec-9-yl) amino) cyclohexyl)-6-methoxy- 2H-indazol-5-yl)-6-(tri- fluoromethyl) pyridinecarboxamide LC-MS: (ESI, m/z): [M + H] .sup.+ = 613.4.

Intermediate 87: N-(6-methoxy-2-(3-(piperidin-4-ylmethyl)-3-azaspiro [5.5]undec-9-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: tert-butyl 9-(6-methoxy-5-nitro-2H-indazol-2-yl)-3-azaspiro[5.5]undecyl-3-carboxylate

[1657] ##STR00547##

[1658] Tert-butyl 9-amino-3-azaspiro[5.5] undecyl-3-carboxylate (498 mg, 1.85 mmol) was added to a solution of 2-azido-4-methoxy-5-nitrobenzaldehyde (416 mg, 1.85 mmol) in toluene (50 mL). The reaction mixture was heated to 110° C. and stirred for reaction overnight. The reaction mixture was concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 600 mg of the target product as a yellow solid.

[1659] LC-MS: (ESI, m/z): [M+H].sup.+=445.2.

Step 2: tert-butyl 9-(5-amino-6-methoxy-2H-indazol-2-yl)-3-azaspiro[5.5]undecyl-3-carboxylate

[1660] ##STR00548##

[1661] Raney-Ni (60 mg) and hydrazine hydrate (85%, 1 mL) were added to a mixture of tert-butyl 9-(6-methoxy-5-nitro-2H-indazol-2-yl)-3-azaspiro [5.5] undecyl-3-carboxylate (600 mg, 1.35 mmol) in ethanol (20 mL). The reaction mixture was stirred at room temperature for 2 hours, filtered, and concentrated under reduced pressure to obtain 450 mg of crude target product as a yellow gel. The crude product can be used for the next step without further purification.

[1662] LC-MS: (ESI, m/z): [M+H].sup.+=415.2.

Step 3: tert-butyl 9-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl)-3-azaspiro[5.5] undecyl-3-carboxylate

[1663] ##STR00549##

[1664] 6-(Trifluoromethyl) picolinic acid (208 mg, 1.09 mmol), HATU (538 mg, 1.42 mmol) and N,N-diisopropylethylamine (554 mg, 4.26 mmol) were added to a mixture of tert-butyl 9-(5-amino-6-methoxy-2H-indazol-2-yl)-3-azaspiro[5.5] undecyl-3-carboxylate (450 mg, 1.09 mmol) in N,N-dimethylformamide (10 mL), and the reaction mixture was stirred at room temperature for 2 hours. The reaction was quenched with water (100 mL) and extracted with ethyl acetate (3×50 mL). The organic layer was collected and washed with saturated saline (150 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by chromatography to obtain 600 mg of the target product as a light yellow solid.

[1665] LC-MS: (ESI, m/z): [M+H].sup.+=588.2.

Step 4: N-(6-methoxy-2-(3-azaspiro[5.5] undec-9-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1666] ##STR00550##

[1667] Trifluoroacetic acid (2 mL) was added to a mixture of tert-butyl 9-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl)-3-azaspiro[5.5]undecyl-3-carboxylate (200 mg, 0.34 mmol) in dichloromethane (5 mL). The reaction mixture was stirred at room temperature for 2 hours, and concentrated under reduced pressure to obtain 270 mg of the desired crude target product as a light yellow solid, which was directly used for the next step.

[1668] LC-MS: (ESI, m/z): [M+H].sup.+=488.3.

Step 5: tert-butyl 4-((9-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl)-3-azaspiro[5.5] undec-3-yl) methyl) piperidine-1-carboxylate

[1669] ##STR00551##

[1670] A reaction mixture of N-(6-methoxy-2-(3-azaspiro[5.5]undec-9-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (270 mg, crude), tert-butyl 4-formylpiperidine-1-carboxylate (130 mg, 0.61 mmol) and sodium triacetoxylborohydride (643 mg, 3.05 mmol) in tetrahydrofuran (10 mL) was stirred and reacted overnight at 60° C. The reaction was quenched with water (50 mL) and extracted with dichloromethane (3×20 mL), and the organic layer was collected, washed with saturated saline (60 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 130 mg of target product as a light yellow solid.

[1671] LC-MS: (ESI, m/z): [M+H].sup.+=685.6.

Step 6: N-(6-methoxy-2-(3-(piperidin-4-ylmethyl)-3-azaspiro [5.5]undec-9-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1672] ##STR00552##

[1673] Trifluoroacetic acid (2 mL) was added to a stirred mixture of tert-butyl 4-((9-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl)-3-azaspiro [5.5] undec-3-yl) methyl) piperidine-1-carboxylate (130 mg, 0.19 mmol) in dichloromethane (5 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated to obtain 220 mg of crude target product as a light yellow solid, and the crude product was directly used for the next step.

[1674] LC-MS: (ESI, m/z): [M+H].sup.+=585.3.

Intermediate 88: tert-butyl 9-(4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl)-3, 9-diazaspiro [5.5] undecyl-3-carboxylate

Step 1: 4-aminocyclohexyl-1-one

[1675] ##STR00553##

[1676] Trifluoroacetic acid (5 mL) was added to a solution of tert-butyl (4-oxocyclohexyl) carbamate (2.13 g, 10 mmol) in dichloromethane (20 mL), and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure to obtain 2.1 g of the crude target product, which can be used for the next step without further purification.

Step 2:4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl-1-one

[1677] ##STR00554##

[1678] 2-Azido-4-methoxy-5-nitrobenzaldehyde (2.22 g, 10 mmol) was added to a reaction mixture of 4-aminocyclohexyl-1-one (crude, 2.1 g) in toluene (50 mL), and the reaction mixture was heated to 100° C. and stirred for reaction overnight, and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 1.5 g of target compound as a yellow solid.

[1679] LC-MS: (ESI, m/z): [M+H].sup.+=290.1

Step 3: tert-butyl 9-(4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl)-3, 9-diazaspiro [5.5] undecyl-3-carboxylate

[1680] ##STR00555##

[1681] A reaction mixture of 4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl-1-one (500 mg, 1.73 mmol), tert-butyl 3,9-diazaspiro [5.5] undecyl-3-carboxylate (484 mg, 1.90 mmol), sodium triacetoxyborohydride (4.00 g, 18.95 mmol) and acetic acid (12 mg, 0.19 mmol) in dichloroethane (100 mL) was refluxed overnight. The reaction mixture was quenched with water (100 mL), and extracted with dichloromethane (3-50 mL). The organic layer was collected, washed with saturated saline (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain the target products P1:150 mg and P2:160 mg as a light yellow solid.

P1: tert-butyl 9-((1r,4r)-4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl)-3, 9-diazaspiro [5.5] undecyl-3-carboxylate

[1682] ##STR00556##

[1683] LC-MS-P1: (ESI, m/z): [M+H].sup.+=528.3.

P2: tert-butyl 9-((1s,4s)-4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl)-3, 9-diazaspiro [5.5] undecyl-3-carboxylate

[1684] ##STR00557##

[1685] LC-MS-P2: (ESI, m/z): [M+H].sup.+=528.3.

Intermediate 89: N-(2-((1s, 4s)-4-(3, 9-diazaspiro [5.5] undec-3-yl) cyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: tert-butyl 9-((1s, 4s)-4-(5-amino-6-methoxy-2H-indazol-2-yl) cyclohexyl)-3, 9-diazaspiro [5.5] undecyl-3-carboxylate

[1686] ##STR00558##

[1687] Palladium/carbon (100 mg) was added to a solution of tert-butyl 9-(4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl)-3, 9-diazaspiro[5.5]undecyl-3-carboxylate (P2, 160 mg, 0.30 mmol) in methanol (10 mL). The resulting mixture was stirred for 4 hours at room temperature in a hydrogen atmosphere. The reaction mixture was filtered through diatomite and the filtrate was concentrated to obtain 140 mg of crude target product as a light yellow solid. The crude mixture was used directly for the next step.

[1688] LC-MS: (ESI, m/z): [M+H].sup.+=498.3.

Step 2: tert-butyl 9-((1s, 4s)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl)-3, 9-diazaspiro[5.5]undecyl-3-carboxylate

[1689] ##STR00559##

[1690] 6-(Trifluoromethyl) picolinic acid (58 mg, 0.30 mmol), HATU (149 mg, 0.39 mmol) and N,N-diisopropylethylamine (117 mg, 0.90 mmol) were added to a stirred solution of tert-butyl 9-((1s, 4s)-4-(5-amino-6-methoxy-2H-indazol-2-yl) cyclohexyl)-3, 9-diazaspiro [5.5]undecyl-3-carboxylate (140 mg, 0.23 mmol) in N,N-dimethylformamide (3 mL). The resulting reaction mixture was stirred for 2 hours at room temperature. The reaction mixture was quenched with water (50 mL), and extracted with ethyl acetate (3-20 mL). The organic layers were collected and washed with saturated saline (60 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by chromatographic column to obtain 90 mg of target product as light yellow solid.

[1691] LC-MS: (ESI, m/z): [M+H].sup.+=671.3.

Step 3: N-(2-((1s, 4s)-4-(3, 9-diazaspiro [5.5] undec-3-yl) cyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1692] ##STR00560##

[1693] Trifluoroacetic acid (2 mL) was added to a solution of tert-butyl 9-((1s, 4s)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl)-3, 9-diazaspiro[5.5] undecyl-3-carboxylate (90 mg, 0.13 mmol) in dichloromethane (5 mL). The resulting reaction mixture was stirred for 2 hours at room temperature. The reaction mixture was concentrated to obtain 120 mg of the crude target product as a light yellow solid.

[1694] LC-MS: (ESI, m/z): [M+H].sup.+=571.3

[1695] The following compounds were prepared with reference to the method of intermediate 89.

TABLE-US-00015 Intermediate Structure Name LC-MS Intermediate 90 [00561] N-(2-((1r,4r)-4-(3, 9-diazaspiro [5.5] undec-3-yl) cyclohexyl)-6-methoxy- 2H-indazol-5-yl)-6-(tri- fluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H].sup.+ = 571.2.

Intermediate 91: N-(6-methoxy-2-((1r,4r)-4-((2-(piperidin-4-yl) ethoxy) methyl) cyclohexyl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: benzyl 4-(2-hydroxyethyl) piperidine-1-carboxylate

[1696] ##STR00562##

[1697] Benzyl carbonyl chloride (5.06 g, 29.8 mmol) was added to a mixture of 2-(piperidin-4-yl) ethyl-1-ol (3.2 g, 24.8 mmol) in saturated sodium bicarbonate aqueous solution (30 mL) and water (30 mL). The reaction mixture was stirred overnight at room temperature, and extracted with ethyl acetate (3×100 mL). The organic layer was collected, washed with saturated saline (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 6.0 g of the target product as a yellow oil.

[1698] LC-MS: (ESI, m/z): [M+H].sup.+=264.3.

Step 2: benzyl 4-(2-(toluenesulfonyloxy) ethyl) piperidine-1-carboxylate

[1699] ##STR00563##

[1700] A mixture of benzyl 4-(2-hydroxyethyl) piperidine-1-carboxylate (6.0 g, 22.8 mmol), p-toluenesulfonyl chloride (5.23 g, 27.4 mmol), triethylamine (6.9 g, 68.4 mmol) and 4-dimethylaminopyridine (0.278 g, 2.28 mmol) in dichloromethane (100 mL) was stirred overnight at room temperature. The reaction mixture was diluted with water (100 mL) and extracted with dichloromethane (3×100 mL). The organic layers were collected and washed with saturated saline (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 8.0 g of the target product as a white solid.

[1701] .sup.1H NMR (400 MHz, DMSO) δ 7.85-7.75 (m, 2H), 7.49 (d, J=8.0 Hz, 2H), 7.43-7.24 (m, 5H), 5.05 (s, 2H), 4.08-4.02 (m, 2H), 3.92 (d, J=13.2 Hz, 2H), 2.68 (s, 2H), 2.42 (s, 3H), 1.55-1.38 (m, 5H), 1.01-0.85 (m, 2H).

Step 3: benzyl 4-(2-(((1r, 4r)-4-((tert-butoxycarbonyl) amino) cyclohexyl) methoxy) ethyl) piperidine-1-carboxylate

[1702] ##STR00564##

[1703] At 0° C., sodium hydride (60%, 210 mg) was added to a solution of tert-butyl ((1r,4r)-4-(hydroxymethyl) cyclohexyl) carbamate (1.0 g, 4.37 mmol) in anhydrous tetrahydrofuran (20 mL), and the reaction mixture was stirred at this temperature for 1 hour, benzyl 4-(2-(toluenesulfonyloxy) ethyl) piperidine-1-carboxylate (2.0 g, 4.80 mmol) was added, the reaction mixture was heated to 60° C. and refluxed overnight. Water (50 mL) was slowly added, extracted with ethyl acetate (100 mL*3), the organic layer was collected, washed with saturated saline (50 mL), concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 1.2 g of target product as a colorless oil.

[1704] LC-MS: (ESI, m/z): [M+H].sup.+=475.3.

Step 4: benzyl 4-(2-(((1r, 4r)-4-aminocyclohexyl) methoxy) ethyl) piperidine-1-carboxylate

[1705] ##STR00565##

[1706] Trifluoroacetic acid (20 mL) was added to a solution of benzyl 4-(2-(((1r, 4r)-4-((tert-butoxycarbonyl) amino) cyclohexyl) methoxy) ethyl) piperidine-1-carboxylate (1.4 g, 2.95 mmol) in dichloromethane (50 mL), and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain 1.4 g of crude target product. It can be used for the next step without further purification.

[1707] LC-MS: (ESI, m/z): [M+H].sup.+=375.3.

Step 5: benzyl 4-(2-(((1r, 4r)-4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl) methoxy) ethyl) piperidine-1-carboxylate

[1708] ##STR00566##

[1709] Benzyl 4-(2-(((1r, 4r)-4-aminocyclohexyl) methoxy)ethyl) piperidine-1-carboxylate (1.4 g, 2.95 mmol) was added to a solution of 2-azido-4-methoxy-5-nitrobenzaldehyde (661 mg, 2.95 mmol) in toluene (50 mL), the reaction mixture was heated to 110° C. and stirred for reaction overnight, concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 0.9 g of target product as a yellow solid.

[1710] LC-MS: (ESI, m/z): [M+H].sup.+=551.2.

Step 6: benzyl 4-(2-(((1r, 4r)-4-(5-amino-6-methoxy-2H-indazol-2-yl) cyclohexyl) methoxy) ethyl) piperidine-1-carboxylate

[1711] ##STR00567##

[1712] Raney-Ni (100 mg) and hydrazine hydrate (85%, 1 mL) were added to a mixture of benzyl 4-(2-(((1r, 4r)-4-(6-methoxy-5-nitro-2H-indazol-2-yl) cyclohexyl) methoxy) ethyl) piperidine-1-carboxylate (570, 1.0 mmol) in ethanol (20 mL), the reaction mixture was stirred for 2 hours at room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain 500 mg of crude target product as a yellow gel. The crude product can be used for the next step without further purification.

[1713] LC-MS: (ESI, m/z): [M+H].sup.+=521.4.

Step 7: benzyl 4-(2-(((1r,4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) methoxy) ethyl) piperidine-1-carboxylate

[1714] ##STR00568##

[1715] 6-(Trifluoromethyl) picolinic acid (184 mg, 0.96 mmol), HATU (474 mg, 1.25 mmol) and N,N-diisopropylethylamine (372 mg, 2.88 mmol) were added to a mixture of benzyl 4-(2-(((1r, 4r)-4-(5-amino-6-methoxy-2H-indazol-2-yl) cyclohexyl) methoxy) ethyl) piperidine-1-carboxylate (500 mg, 0.96 mmol) in N,N-dimethylformamide (10 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction was quenched with water (20 mL). The resulting solution was extracted with ethyl acetate (3×50 mL). The organic layers were collected and washed with saturated saline (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by column chromatography to obtain 700 mg of target product as a yellow solid, .sup.1H-NMR showed that the target product contained N,N-dimethylformamide.

[1716] LC-MS: (ESI, m/z): [M+H].sup.+=694.2.

Step 8: N-(6-methoxy-2-((1r,4r)-4-((2-(piperidin-4-yl) ethoxy) methyl) cyclohexyl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1717] ##STR00569##

[1718] Palladium carbon (10%, 70 mg) was added to a mixture of benzyl 4-(2-(((1r,4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) methoxy) ethyl) piperidine-1-carboxylate (700 mg, 0.96 mmol) in ethyl acetate (20 mL), the reaction mixture was stirred overnight at room temperature in a hydrogen atmosphere, filtered, the solid was washed with ethyl acetate (20 mL), and the organic layer was collected, concentrated under reduced pressure to obtain 550 mg of crude target product. The crude product can be used for the next step without further purification.

[1719] LC-MS: (ESI, m/z): [M+H].sup.+=560.3.

Intermediate 92: N-(2-((1r,4r)-4-(([1,4′-bipiperidin]-4-yl (methyl) amino) methyl) cyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: tert-butyl 4-(((1r,4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) methyl) (methyl) amino)-[1,4′-bipiperidine]-1′-carboxylate

[1720] ##STR00570##

[1721] N-(6-methoxy-2-((1r,4r)-4-((methyl (piperidin-4-yl) amino) methyl) cyclohexyl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (280 mg, 0.40 mmol) and tert-butyl 4-oxopiperidine-1-carboxylate (88 mg, 0.44 mmol) were dissolved with 1,2-dichloroethane (20 mL), the temperature was increased to 70° C., three drops of acetic acid were added, sodium triacetoxyborohydride (844 mg, 4.0 mmol) was divided into five parts, one part was added to the reaction solution every 1 hour, and stirred overnight at 70° C. The reaction solution was cooled to room temperature, added with water (30 mL), extracted with dichloromethane (3×20 mL), and the organic phase was washed with saturated saline (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain 70 mg of target product as a light yellow solid.

[1722] LC-MS: (ESI, m/z): [M+H].sup.+=728.4.

Step 2: N-(2-((1r,4r)-4-(([1,4′-bipiperidin]-4-yl (methyl) amino) methyl) cyclohexyl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1723] ##STR00571##

[1724] Tert-butyl 4-((((1r,4r)-4-(6-methoxy-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) cyclohexyl) methyl) (methyl) amino)-[1,4′-biperidine]-1′-carboxylate (70 mg, 0.096 mmol) was dissolved with dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added and stirred at room temperature for 2 hours, directly concentrated under reduced pressure to obtain 100 mg of crude target product as a yellow solid, which was directly used for the next reaction without purification.

[1725] LC-MS: (ESI, m/z): [M+H].sup.+=628.3.

Intermediate 93: N-(2-(1-((3-azaspiro [5.5] undec-9-yl) methyl) piperidin-4-yl)-6-(difluoromethoxy)-2H-indazol-5-yl)-6-(trifluoromethyl)) pyridinecarboxamide

Step 1: tert-butyl 4-(6-hydroxy-5-nitro-2H-indazol-2-yl) piperidine-1-carboxylate

[1726] ##STR00572##

[1727] Tert-butyl 4-(6-methoxy-5-nitro-2H-indazol-2-yl) piperidine-1-carboxylate (5.0 g, 13.29 mmol) was dissolved in hydrogen bromide aqueous solution (48%, 60 mL), heated to 110° C., stirred for 48 hours, adjusted the pH value of the reaction solution to 9-10 with sodium hydroxide aqueous solution, and ditert-butyl dicarbonate (3.73 g, 13.29 mmol) and 4-dimethylaminopyridine (324 mg, 2.65 mmol) were added, stirred overnight at room temperature, extracted with ethyl acetate (3×100 mL), the organic phases was washed with saturated saline (300 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain 530 mg of target compound as an orange-red solid.

[1728] LC-MS: (ESI, m/z): [M+H].sup.+=363.1.

[1729] .sup.1H NMR (400 MHz, DMSO) δ 10.47 (s, 1H), 8.62 (s, 1H), 8.42 (s, 1H), 7.02 (s, 1H), 4.68 (tt, J=11.4, 4.0 Hz, 1H), 4.18-3.98 (m, 2H), 2.94 (m, 2H), 2.10 (d, J=10.1 Hz, 2H), 1.92 (qd, J=12.3, 4.3 Hz, 2H), 1.43 (s, 9H).

Step 2: tert-butyl 4-(6-(difluoromethoxy)-5-nitro-2H-indazol-2-yl) piperidine-1-carboxylate

[1730] ##STR00573##

[1731] Sodium hydroxide (1.17 g, 29.2 mmol) was dissolved in water (10 mL) and acetonitrile (10 mL), cooled to −20° C., tert-butyl 4-(6-hydroxy-5-nitro-2H-indazol-2-yl) piperidine-1-carboxylate (530 mg, 1.46 mmol) was added, diethyl (bromodifluoromethyl) phosphonate (782 mg, 2.92 mmol) was slowly added, stirred at −20° C. for 3h, extracted with ethyl acetate (3×30 mL), the organic phase was washed with saturated saline (90 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified to obtain 420 mg of the target product.

[1732] LC-MS: (ESI, m/z): [M−55] *=357.2.

[1733] .sup.1H NMR (400 MHz, DMSO) δ 8.84 (s, 1H), 8.68 (s, 1H), 7.64 (s, 1H), 7.31 (t, J=73.2 Hz, 1H), 4.89-4.75 (m, 1H), 4.11 (d, J=12.0 Hz, 2H), 2.97 (s, 2H), 2.13 (d, J=10.5 Hz, 2H), 2.02-1.89 (m, 3H), 1.44 (s, 9H).

Step 3: tert-butyl 4-(5-amino-6-(difluoromethoxy)-2H-indazol-2-yl) piperidine-1-carboxylate

[1734] ##STR00574##

[1735] Tert-butyl 4-(6-(difluoromethoxy)-5-nitro-2H-indazol-2-yl) piperidine-1-carboxylate (420 mg, 1.02 mmol) was added to ethanol (20 mL), Raney-Ni (0.5 mL) and hydrazine hydrate (0.5 mL) were added under stirring, stirred for 2 hours at room temperature, filtered, and the filter cake was washed twice with ethanol (5 mL×2), concentrated under reduced pressure to obtain 380 mg of the target compound, which can be directly used in the next reaction without purification.

[1736] LC-MS: (ESI, m/z): [M+H].sup.+=383.1

Step 4: tert-butyl 4-(6-(difluoromethoxy)-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidine-1-carboxylate

[1737] ##STR00575##

[1738] Tert-butyl 4-(5-amino-6-(difluoromethoxy)-2H-indazol-2-yl) piperidine-1-carboxylate (380 mg, 0.99 mmol) was dissolved in N,N-dimethylformamide (5 mL), 6-(trifluoromethyl) picolinic acid (229 mg, 1.2 mmol), HATU (494 mg, 1.3 mmol) and N,N-diisopropylethylamine (387 mg, 3.0 mmol) were added, stirred at room temperature for 2 hours, water (20 mL) was added, filtered, the filter cake was washed with water (10 mL×2) twice to obtain crude product, and the crude product was purified by column chromatography to obtain 400 mg of target compound.

[1739] LC-MS: (ESI, m/z): [M−55].sup.+=556.0.

[1740] .sup.1H NMR (400 MHz, DMSO) δ 10.51 (s, 1H), 8.69 (s, 1H), 8.46 (d, J=7.4 Hz, 1H), 8.41 (t, J=7.8 Hz, 1H), 8.35 (s, 1H), 8.22 (dd, J=7.7, 1.0 Hz, 1H), 7.15 (s, 1H), 4.39 (ddd, J=11.7, 8.0, 3.8 Hz, 1H), 3.97 (d, J=14.3 Hz, 5H), 3.04 (s, 1H), 2.76 (d, J=6.1 Hz, 4H), 2.17 (d, J=10.1 Hz, 2H), 2.00 (d, J=8.7 Hz, 4H), 1.93-1.84 (m, 2H), 1.73 (s, 1H), 1.39 (d, J=15.8 Hz, 11H), 1.21 (dt, J=14.3, 9.8 Hz, 2H).

Step 5: N-(6-(difluoromethoxy)-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1741] ##STR00576##

[1742] Tert-butyl 4-(6-(difluoromethoxy)-5-(6-(trifluoromethyl) pyridinecarboxamide)-2H-indazol-2-yl) piperidine-1-carboxylate (200 mg, 0.36 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added and stirred at room temperature for 2 hours, directly concentrated under reduced pressure to obtain 220 mg of crude target product, which was directly used for the next reaction without purification.

[1743] LC-MS: (ESI, m/z): [M+H].sup.+=456.1.

Step 6: tert-butyl 9-((4-(6-(difluoromethoxy)-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5]undecyl-3-carboxylate

[1744] ##STR00577##

[1745] N-(6-(difluoromethoxy)-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (220 mg) and tert-butyl 9-formyl-3-azaspiro[5.5] undecyl-3-carboxylate (109 mg, 0.38 mmol) were dissolved in tetrahydrofuran (10 mL), stirred at room temperature, sodium triacetoxyborohydride (371 mg, 1.76 mmol) was added and stirred at room temperature for 2 hours, water (30 mL) was added, extracted with dichloromethane (3×30 mL), the organic phase was washed with saturated saline (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain 100 mg of the target product.

[1746] LC-MS: (ESI, m/z): [M+H].sup.+=721.7.

Step 7: N-(2-(1-((3-azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6-(difluoromethoxy)-2H-indazol-5-yl)-6-(trifluoromethyl)) pyridinecarboxamide

[1747] ##STR00578##

[1748] Tert-butyl 9-((4-(6-(difluoromethoxy)-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5]undecyl-3-carboxylate (100 mg, 0.138 mmol) was dissolved with dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added and stirred at room temperature for 2 hours, and it was directly concentrated under reduced pressure to obtain 120 mg of target crude product, which was directly used for the next reaction without purification.

[1749] LC-MS: (ESI, m/z): [M+H].sup.+=621.2.

Intermediate 94: tert-butyl 4-(4-aminobutyl) piperidine-1-carboxylate

Step 1: tert-butyl 4-(4-(1, 3-dioxoisoindolin-2-yl) butyl) piperidine-1-carboxylate

[1750] ##STR00579##

[1751] Tert-butyl 4-(4-hydroxybutyl) piperidine-1-carboxylate (1.2 g, 4.73 mmol), phthalimide (0.82 g, 5.52 mmol) and triphenylphosphine (1.45 g, 5.52 mmol) were dissolved in anhydrous tetrahydrofuran (25 mL), the reaction solution was stirred at room temperature under nitrogen protection for 10 minutes, and then cooled to 0° C., diethyl azodicarate (0.87 mL, 5.52 mmol) was slowly added dropwise to the reaction solution at 0° C., the ice bath was removed, stirred at room temperature for 7 hours, the solvent was removed by concentration under reduced pressure, and the concentrate was purified by column chromatography (20% ethyl acetate/petroleum ether) to obtain 1.5 g of target compound as a white solid.

[1752] LC-MS: (ESI, m/z): [M−99].sup.+=387.2.

Step 2: tert-butyl 4-(4-aminobutyl) piperidine-1-carboxylate

[1753] ##STR00580##

[1754] Tert-butyl 4-(4-hydroxybutyl) piperidine-1-carboxylate (1.5 g, 3.88 mmol) and hydrazine hydrate (5 mL) were added to ethanol (20 mL), heated and refluxed overnight, cooled to room temperature, filtered, the filter cake was washed twice with ethanol (5 mL×2), and concentrated under reduced pressure to obtain 1.0 g of target compound as a colorless oil, which can be directly used for the next reaction without purification.

[1755] .sup.1H NMR (400 MHz, DMSO) δ 3.95 (t, J=27.6 Hz, 2H), 2.64 (s, 2H), 2.57-2.52 (m, 2H), 1.60 (d, J=12.4 Hz, 2H), 1.48-1.21 (m, 14H), 1.17 (dd, J=13.9, 6.3 Hz, 2H), 0.92 (qd, J=12.5, 4.2 Hz, 2H).

Intermediate 95: N-(2-(1-((3-azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6-(2-methoxyethoxy)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: 5-nitro-2-(piperidin-4-yl)-2H-indazol-6-ol

[1756] ##STR00581##

[1757] Tert-butyl 4-(6-hydroxy-5-nitro-2H-indazol-2-yl) piperidine-1-carboxylate (1.4 g, crude product) was dissolved in 4M hydrochloric acid-1,4-dioxane (20 mL), stirred for 3 hours at room temperature, and concentrated directly under reduced pressure to obtain 800 mg of crude target product as a light yellow solid. The crude product was directly used for the next reaction without purification.

[1758] LC-MS: (ESI, m/z): [M+H].sup.+=263.1.

Step 2: tert-butyl 9-((4-(6-hydroxy-5-nitro-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5] undecyl-3-carboxylate

[1759] ##STR00582##

[1760] 5-Nitro-2-(piperidin-4-yl)-2H-indazol-6-ol (500 mg) and tert-butyl 9-formyl-3-azaspiro [5.5] undecyl-3-carboxylate (536 mg, 1.9 mmol) were dissolved in tetrahydrofuran (20 mL), stirred at room temperature, sodium triacetoxyborohydride (1.2 g, 5.7 mmol) was added and stirred at room temperature for 2 hours, water (50 mL) was added, extracted with dichloromethane (3×30 mL), the organic phase was washed with saturated saline (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain 440 mg of target compound as a yellow solid.

[1761] LC-MS: (ESI, m/z): [M+H].sup.+=528.2.

Step 3: tert-butyl 9-((4-(6-(2-methoxyethoxy)-5-nitro-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5] undecyl-3-carboxylate

[1762] ##STR00583##

[1763] Tert-butyl 9-((4-(6-hydroxy-5-nitro-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5] undecyl-3-carboxylate (440 mg, 0.8 mmol) and 1-bromo-2-methoxyethane (1.15 g, 8.0 mmol) were dissolved with acetonitrile (20 mL), anhydrous potassium carbonate (331 mg, 2.4 mmol) was added and heated to 80° C., stirred for 2 hours, water (50 mL) was added, extracted with ethyl acetate (3×30 mL), organic phase was washed with saturated saline (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain 110 mg of target compound as a yellow solid.

[1764] LC-MS: (ESI, m/z): [M+H].sup.+=586.5.

Step 4: tert-butyl 9-((4-(5-amino-6-(2-methoxyethoxy)-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5] undecyl-3-carboxylate

[1765] ##STR00584##

[1766] Tert-butyl 9-((4-(6-(2-methoxyethoxy)-5-nitro-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5] undecyl-3-carboxylate (110 mg, 0.18 mmol) was added to ethanol (10 mL), Raney-Ni (0.5 mL) and hydrazine hydrate (0.5 mL) were added under stirring, stirred for 2 hours at room temperature, filtered, and the filter cake was washed twice with ethanol (5 mL×2), and concentrated under reduced pressure to obtain 100 mg of target compound as a yellow solid, which can be directly used for the next reaction without purification.

[1767] LC-MS: (ESI, m/z): [M+H].sup.+=556.5.

Step 5: tert-butyl 9-((4-(6-(2-methoxyethoxy))-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5]undecyl-3-carboxylate

[1768] ##STR00585##

[1769] Tert-butyl 9-((4-(5-amino-6-(2-methoxyethoxy)-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5] undecyl-3-carboxylate (100 mg, 0.18 mmol) was dissolved in N,N-dimethylformamide (5 mL), 6-(trifluoromethyl) picolinic acid (41 mg, 0.21 mmol), HATU (89 mg, 0.23 mmol) and N,N-diisopropylethylamine (70 mg, 0.54 mmol) were added, stirred for 2 hours at room temperature, water (20 mL) was added, extracted with ethyl acetate (3×20 mL), the organic phase was washed with saturated saline (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain 80 mg of target product as a yellow solid.

[1770] LC-MS: (ESI, m/z): [M+H].sup.+=729.1.

[1771] .sup.1H NMR (400 MHz, DMSO) δ 10.57 (s, 1H), 8.74 (s, 1H), 8.48 (d, J=7.8 Hz, 1H), 8.41 (t, J=7.8 Hz, 1H), 8.37 (s, 1H), 8.22 (d, J=7.7 Hz, 1H), 7.17 (s, 1H), 4.37 (m, 1H), 4.32-4.26 (m, 2H), 3.86-3.77 (m, 2H), 3.27 (m, 5H), 2.94 (m, 2H), 2.17 (m, 1=5.6 Hz, 2H), 2.07 (m, J=5.3 Hz, 6H), 1.64 (d, J=7.7 Hz, 2H), 1.56 (d, J=9.3 Hz, 2H), 1.53-1.32 (m, 14H), 1.22 (d, J=7.0 Hz, 2H), 1.12-0.98 (m, 4H).

Step 6: N-(2-(1-((3-azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6-(2-methoxyethoxy)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1772] ##STR00586##

[1773] Tert-butyl 9-((4-(5-amino-6-(2-methoxyethoxy)-2H-indazol-2-yl) piperidin-1-yl) methyl)-3-azaspiro[5.5] undecyl-3-carboxylate (80 mg, 0.110 mmol) was dissolved with dichloromethane (5 mL), trifluoroacetic acid (2 mL) was added and stirred at room temperature for 2 hours, directly concentrated under reduced pressure to obtain 105 mg of crude target compound, which was directly used for the next reaction without purification.

[1774] LC-MS: (ESI, m/z): [M+H].sup.+=629.1.

[1775] The following intermediates were prepared with reference to the method of intermediate 95.

TABLE-US-00016 Intermediate Structure Name LC-MS Intermediate 96 [00587] N-(2-(1-((3-azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)- 6-(oxetan-3-yloxy)-2H- indazol-5-yl)- 6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H] .sup.+ = 627.3

Intermediate 97: benzyl 4-(((1r,4r)-4-((tert-butoxycarbonyl) amino) cyclohexyl) methoxy) methyl) piperidine-1-carboxylate

Step 1: tert-butyl ((1r,4r)-4-((pyridin-4-ylmethoxy) methyl) cyclohexyl) carbamate

[1776] ##STR00588##

[1777] Sodium hydride (211 mg, 8.8 mmol) was added to tert-butyl ((1r,4r)-4-(hydroxymethyl) cyclohexyl) carbamate (1.0 g, 4.4 mmol) in tetrahydrofuran (20 mL) at 0° C., stirred at 0° C. for 30 minutes, continued stirring at room temperature for 30 minutes, 4-bromomethylpyridine (1.1 g, 4.4 mmol) was added, and stirred at room temperature overnight, water (30 mL) was added, extracted with ethyl acetate (3×50 mL), the organic phase was washed with saturated saline (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography (50% EA/PE) to obtain 1.0 g of target product as a white solid.

[1778] LC-MS: (ESI, m/z): [M+H].sup.+=321.1.

Step 2: tert-butyl ((1r,4r)-4-((piperidin-4-ylmethoxy) methyl) cyclohexyl) carbamate

[1779] ##STR00589##

[1780] Tert-butyl ((1r,4r)-4-((pyridin-4-ylmethoxy) methyl) cyclohexyl) carbamate (1.0 g, 3.1 mmol) was dissolved in isopropanol (30 mL) and water (35 mL), palladium/carbon (0.5 g) was added, stirred at 75° C. for 72 hours in hydrogen environment, filtered, concentrated and dried to obtain 1.0 g of crude target product as a white solid.

[1781] LC-MS: (ESI, m/z): [M+H].sup.+=327.2.

Step 3: benzyl 4-((((1r,4r)-4-((tert-butoxycarbonyl) amino) cyclohexyl) methoxy) methyl) piperidine-1-carboxylate

[1782] ##STR00590##

[1783] Saturated sodium bicarbonate solution (30 mL) and benzyl chloroformate (680 mg, 3.98 mmol) were added to tert-butyl ((1r,4r)-4-((piperidin-4-ylmethoxy) methyl) cyclohexyl) carbamate (1.0 g, 3.06 mmol) in ethyl acetate (30 mL), stirred overnight at room temperature, water (30 mL) was added, extracted with ethyl acetate (3×50 mL), the organic phase was washed with saturated saline (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain 400 mg of target product as a white solid.

[1784] LC-MS: (ESI, m/z): [M+H].sup.+=461.2.

[1785] The following intermediates were prepared with reference to the method of intermediate 91.

TABLE-US-00017 Intermediate Structure Name LC-MS Intermediate 98 [00591] N-(6-methoxy- 2-((1r,4r)-4-((piperidin-4- ylmethoxy) methyl) cyclohexyl)-2H-indazol-5- yl)-6-(trifluoromethyl) pyridinecarboxamide (ESI, m/z): [M + H] .sup.+ = 546.2

Example 1: N-(2-(1-(2-(9-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) ethyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl)-2-pyridinecarboxamide

[1786] ##STR00592##

[1787] N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (95 mg, 0.226 mmol) was added to a mixture of 2-(9-(3-(2, 4-dioxo tetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-3,9-diazaspiro[5.5] undec-3-yl) acetaldehyde (100 mg, 0.226 mmol) and sodium cyanoborohydride (28 mg, 0.452 mmol) in methanol/acetic acid (4 mL, 10:1). The mixture was stirred at room temperature for 4h, concentrated under reduced pressure, and the concentrate was purified by preparative HPLC to obtain 11.23 mg of the target compound as a light yellow solid.

[1788] LC-MS: (ESI, m/z): [M+H].sup.+=846.3

[1789] 1H NMR (400 MHz, MeOD) δ 8.85 (s, 1H), 8.50-8.42 (m, 2H), 8.32 (t, J=7.9 Hz, 1H), 8.07 (d, J=7.8 Hz, 1H), 7.49-7.39 (m, 2H), 7.20 (d, J=8.6 Hz, 1H), 7.13 (s, 1H), 4.94 (s, 1H), 4.09 (s, 3H), 3.97 (s, 1H), 3.93 (s, 3H), 3.82-3.36 (m, 16H), 3.26-3.18 (m, 1H), 2.81 (t, J=6.7 Hz, 2H), 2.63 (m, J=35.7, 11.9 Hz, 4H), 2.12-1.52 (m, 8H).

[1790] The following compounds were prepared with reference to the method of Example 1:

TABLE-US-00018 Example Structure Name LC-MS/NMR Example 2 [00593] N-(2-(1-(3-(9-(3-(2, 4-dioxotetrahydropyrimi- din-1 (2H)-yl)-4-methoxyben- zoyl)-3,9-diazaspiro [5.5] undec-3-yl) propyl) piperidin-4-yl)-6-methoxy- 2H-indazol-5-yl)-6-(tri- fluoromethyl)-2-pyridine carboxamide LC-MS: (ESI, m/z): [M + H].sup.+ = 860.4. .sup.1HNMR(400 MHz, DMSO) δ 7.98 (s, 1H), 7.69 (d, J = 7.9 Hz, 1H), 7.52 (t, J = 7.8 Hz, 1H), 7.42 (s, 1H), 7.26 (d, J = 7.8 Hz, 1H), 6.66 (m, J = 8.5, 2.0 Hz, 1H), 6.60 (d, J = 2.0 Hz, 1H), 6.40 (d, J = 8.6 Hz, 1H), 6.29 (s, 1H), 3.70-3.59 (m, 1H), 3.27 (s, 3H), 3.13 (s, 3H), 2.94 (m, 4H), 2.74 (s, 2H), 2.35 (d, J = 7.8 Hz, 2H), 2.02 (t, J = 6.7 Hz, 2H), 1.71 (m, J = 23.2, 16.2 Hz, 8H), 1.46 (t, J = 10.2 Hz, 6H), 1.01 (s, 2H), 0.87 (s, 4H), 0.81-0.66 (m, 4H). Example 3 [00594] N-(2-(1-(4-(9-) 3-(2, 4-dioxotetrahydropyrimi- din-1 (2H)-yl)-4-methoxybenzo- yl)-3,9-diazaspiro [5.5] undec-3-yl) butyl) piperidin-4-yl)-6-methoxy- 2H-indazol-5-yl)-6-(tri- fluoromethyl)-2-pyridine carboxamide LC-MS: (ESI, m/z): [M + H] .sup.+ = 874.2 .sup.1HNMR(400 MHz, MeOD) δ 8.76 (s, 1H), 8.47 (d, J = 7.6 Hz, 1H), 8.30 (t, J = 7.7 Hz, 1H), 8.21 (s, 1H), 8.05 (d, J = 7.5 Hz, 1H), 7.47-7.37 (m, 2H), 7.18 (d, J = 8.5 Hz, 1H), 7.07 (s, 1H), 4.42 (s, 1H), 4.05 (s, 3H), 3.92 (s, 3H), 3.72 (s, 4H), 3.52 (s, 2H), 3.12 (s, 2H), 2.80 (t, J = 6.5 Hz, 2H), 2.50 (d, J = 23.1 Hz, 7H), 2.24 (s, 6H), 1.61 (d, J = 22.7 Hz, 13H).

Example 4: N-(2-(1-(5-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzamido) pentyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1791] ##STR00595##

[1792] A mixture of N-(2-(1-(5-aminopentyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (160 mg, 0.317 mmol), 3-(2, 4-dioxotetrahydropyrimidin-1(2H)-yl)-4-methoxybenzoic acid (26 mg, 0.0985 mmol), HATU (49 mg, 0.129 mmol) and N,N-diisopropylethylamine (64 mg, 0.496 mmol) in N,N-dimethylformamide (3 mL was stirred for 3 h at room temperature. The reaction solution was poured into water (20 mL) and extracted with ethyl acetate (3×20 mL), the organic phase was collected, washed with water (2×50 mL) and saline (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by preparative HPLC to obtain 19.97 mg of target product as a light yellow solid.

[1793] LC-MS: (ESI, m/z): [M+H]-751.2.

[1794] 1H NMR (400 MHz, DMSO) δ 10.50 (s, 1H), 10.35 (s, 7H), 8.69 (s, 1H), 8.49-8.32 (m, 4H), 8.22 (d, J=7.6 Hz, JH), 7.86 (d, J=8.8 Hz, 1H), 7.79 (d, J=6.2 Hz, 1H), 7.18 (d, J=8.8 Hz, 2H), 4.45-4.30 (m, 1H), 3.98 (s, 3H), 3.85 (s, 3H), 3.62-3.55 (d 2H), 3.29-3.23 (d, J=6.2 Hz, 2H), 3.05-2.95 (m, 2H), 2.72-2.65 (m, 2H), 2.38-2.29 (m, 2H), 2.15-1.98 (m, 6H), 1.60-1.44 (m, 4H), 1.38-1.28 (in, 2H).

[1795] The following compounds were prepared with reference to the method of Example 4:

TABLE-US-00019 Example Structure Name LC-MS/NMR Example 5 [00596] N-(2-(1-(2-(2-(3-(2, 4-dioxotetrahydropyrim- idin-1 (2H)-yl)-4-methoxybenz- amido) ethoxy) ethyl) piperidin-4-yl)-6-me- thoxy-2H-indazol-5-yl)-6- (trifluoromethyl) pyridinecarboxamide LC-MS: (ESI, m/z): [M + H] .sup.+ = 753.2 1H NMR (400 MHz, DMSO) δ 10.50 (s, 1H), 10.36 (s, 1H), 8.68 (s, 1H), 8.50-8.30 (m, 4H), 8.22 (d J = 7.6 Hz, 1H), 7.91- 7.85 (m, 1H), 7.80 (d, J = 2.0 Hz, 1H), 7.17 (d, J = 9.8 Hz, 2H), 4.37-4.27 (m, 1H), 3.98 (s, 3H), 3.84 (s, 3H), 3.58- 3.53 (m, 6H), 3.45- 3.38 (m, 2H), 3.00 (d, J = 11.3 Hz, 2H), 2.71-2.62 (m, 2H), 2.58 2.51 (m, 2H), 2.25-2.13 (m, 2H), 2.11-1.95 (m, 4H). Example 6 [00597] N-(2-(1-(2-(1-(3-(2, 4-dioxotetrahydropyrim- idin-1 (2H)-yl)-4-methoxyben- zoyl) piperidin-4-yl) ethyl) piperidin-4-yl)-6-me- thoxy-2H-indazol-5-yl)- 6-(trifluoromethyl) pyridinecarboxamide LC-MS: (ESI, m/z): [M + H] .sup.+ = 777.3. .sup.1H NMR (400 MHz, CDCl3) δ 10.71 (s, 1H), 8.82 (s, 1H), 8.50 (d, J = 7.8 Hz, 1H), 8.12 (t, J = 7.8 Hz, 1H), 7.95- 7.82 (m, 2H), 7.59 (s, 1H), 7.45-7.34 (m, 2H), 7.10-6.96 (m, 2H), 4.62-4.50 (m, 1H), 4.04 (s, 3H), 3.90 (s, 3H), 3.76-3.68 (m, 2H), 3.45-3.34 (m, 2H), 2.87-2.79 (m, 2H), 2.82-2.73 (m, 4H), 2.48-2.37 (m, 8H), 1.75-1.69 (m, 5H), 1.30-1.10 (m, 2H).

Example 7: N-(2-(1-(2-(1-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl) piperidin-4-yl) ethyl) piperidin-4-yl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1796] ##STR00598##

[1797] STAB (71 mg, 0.33 mmol) was added to a mixture of 2-(1-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl) piperidin-4-yl) acetaldehyde (80 mg, 0.22 mmol) and N-(6-(2-hydroxyprop-2-yl)-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (50 mg, 0.11 mmol) in tetrahydrofuran (10 mL). The reaction solution was stirred at room temperature for 2 hours. Water (10 mL) and ethyl acetate (10 mL) were added. The water layer was separated and extracted with ethyl acetate (10 mL/2). The organic layers were combined, washed with saline (10 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by preparative HPLC to obtain 9.8 mg of target product as a yellow solid.

[1798] LC-MS: (ES, m/z): [M+H].sup.+=805.3

[1799] .sup.1H NMR (400 MHz, MeOD) δ 8.72 (s, 1H), 8.47 (d, J=7.8 Hz, 2H), 8.32-8.24 (m, 2H), 8.02 (d, J=7.8 Hz, 1H), 7.67 (s, 1H), 7.45 (dd, J=8.5, 2.1 Hz, 1H), 7.39 (d, J=2.0 Hz, 1H), 7.19 (d, J=8.6 Hz, 1H), 5.02-4.96 (m, 1H), 4.59-4.56 (m, 2H), 3.92 (s, 3H), 3.73 (s, 2H), 2.81 (t, J=6.7 Hz, 3H), 2.73-2.69 (m, 2H), 2.53-2.49 (m, 2H), 2.33-2.29 (m, 4H), 1.95-1.55 (m, 12H), 1.29-1.25 (m, 2H).

[1800] The following examples were prepared with reference to the method of example 7:

TABLE-US-00020 Example structure Name LC-MS/NMR Example 8 [00599] N-(2-(1-(2-(2-(2-(3-(2, 4-dioxotetrahydropyrimidin- 1 (2H)-yl)-4-methoxybenz- amido) ethoxy) ethoxy) ethyl) piperidin-4-yl)-6-methoxy- 2H-indazol-5-yl)-6-(trifluoro- methyl) pyridin- carboxamide LC-MS: (ESI, m/z): [M + H].sup.+ = 797.2 .sup.1H-NMR: 1H NMR (400 MHz, ) δ 10.51 (s, 1H), 10.36 (s, 1H), 8.69 (s, 1H), 8.47- 8.36 (m, 3H), 8.22 (d, J = 7.8 Hz, 2H), 7.87 (dd, J = 8.6, 2.2 Hz, 1H), 7.80 (d, J = 2.2 Hz, 1H), 7.17 (t, J = 4.2 Hz, 2H), 4.39- 4.31 (m, 1H), 3.98 (s, 3H), 3.84 (s, 3H), 3.60- 3.53 (m, 10H), 3.44- 3.40 (m, 2H), 3.00 (d, J = 11.3 Hz, 2H), 2.68 (t, J = 6.0 Hz, 2H), 2.53 (d, J = 5.8 Hz, 2H), 2.18 (dd, J = 16.4, 9.2 Hz, 2H), 2.11- 1.99 (m, 4H)

Example 9: N-(2-(1-(5-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxy-N-methylbenzamido) pentyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1801] ##STR00600##

[1802] 3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxy-N-methyl-N-(5-oxopentyl) benzamide (100 mg, 0.27 mmol), dichloroethane (2 mL), methanol (2 mL), N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (113.13 mg, 0.27 mmol), acetic acid (1 drop) and sodium triacetate borohydride (114.48 mg, 0.54 mmol) were added to a 25 mL round bottom flask, and the reaction mixture was stirred at room temperature overnight. 10 mL of water was added to quench the reaction, extracted with dichloromethane (2-20 mL), and the organic layers were combined and washed with 20 mL saturated saline, e dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was purified by preparative HPLC to obtain 36.5 mg of target product as a white solid.

[1803] LC-MS: (ES, m/z): [M+H].sup.+=765.4

[1804] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.51 (s, 1H), 10.34 (s, 1H), 8.69 (s, 1H), 8.49-8.38 (m, 2H), 8.36 (s, 1H), 8.22 (d, J=8.6 Hz, 2H), 7.44-7.27 (m, 2H), 7.16 (d, J=8.4 Hz, 2H), 4.39-4.35 (m, 1H), 3.98 (s, 3H), 3.84 (s, 3H), 3.59 (t, J=6.6 Hz, 3H), 2.97-2.90 (m, 6H), 2.68 (t, J=6.1 Hz, 2H), 2.35-2.31 (m, 2H), 2.11-2.05 (m, 6H), 1.64-1.16 (m, 6H).

Example 10: N-(2-(1-(2-(9-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) ethyl) piperidin-4-yl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1805] ##STR00601##

[1806] 2-(9-(3-(2, 4-Dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) acetaldehyde (150 mg, 0.336 mmol) was added to a mixture of N-(6-(2-hydroxyprop-2-yl)-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (50 mg, 0.112 mmol) and sodium cyanoborohydride (21 mg, 0.336 mmol) in methanol/acetic acid (4 mL, 10:1). The reaction mixture was stirred at room temperature for 4h. Then it was concentrated under reduced pressure, and the concentrate was purified by preparative HPLC to obtain 26.96 mg of the crude target product.

[1807] LC-MS: (ESI, m/z): [M+H].sup.+=874.4.

[1808] .sup.1H NMR (400 MHz, DMSO) δ 12.37 (s, 1H), 10.34 (s, 1H), 8.71 (s, 1H), 8.46-8.34 (m, 3H), 8.16 (dd, J=7.8, 0.8 Hz, 1H), 7.57 (s, 1H), 7.43-7.26 (m, 2H), 7.15 (d, J=8.6 Hz, 1H), 5.95 (s, 1H), 4.44-4.42 (m, 1H), 3.84 (s, 3H), 3.61-3.35 (m, 6H), 3.01 (d, J=10.5 Hz, 2H), 2.68 (t, J=6.6 Hz, 2H), 2.49-2.31 (m, 8H), 2.21-2.03 (m, 6H), 1.62 (s, 6H), 1.48-1.27 (m, 8H).

[1809] The compounds in the following table were prepared with reference to the method of Example 10.

TABLE-US-00021 Example 11 [00602] N-(2-(1-(2-(9-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) propyl) piperidin-4-yl)-6-(2-hydroxyprop- 2-yl)-2H-indazol-5-yl)-6-(trifluoro- methyl) pyridinecarboxamide LC-MS: (ESI, m/z): [M + H].sup.+ = 888.4 .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.33 (s, 1H), 8.71 (s, 1H), 8.45 (d, J = 7.7 Hz, 1H), 8.37 (dd, J = 14.9, 7.1 Hz, 2H), 8.28 (s, 2H), 8.16 (d, J = 7.7 Hz, 1H), 7.57 (s, 1H), 7.41- 7.28 (m, 2H), 7.15 (d, J = 8.6 Hz, 1H), 5.94 (s, 1H), 4.47-4. 43 (m, 1H), 3.84 (s, 3H), 3.60 (t, J = 6.6 Hz, 4H), 3.02- 2.97 (m, 2H), 2.68 (t, J = 6.3 Hz, 2H), 2.35 (d, J = 5.7 Hz, 8H), 2.09 (d, J = 5.7 Hz, 6H), 1.62 (s, 8H), 1.46 (d, J = 29.9 Hz, 8H). Example 12 [00603] N-(2-(1-(2-(7-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-2, 7-diazaspiro [3.5] non-2-yl) ethyl) piperidin-4-yl)-6-(2-hydroxyprop- 2-yl)-2H-indazol-5-yl)-6-(trifluoro- methyl) pyridinecarboxamide LC-MS: (ESI, m/z): [M + H] .sup.+ = 846.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.34 (s, 1H), 8.71 (s, 1H), 8.45 (d, J = 7.8 Hz, 1H), 8.37 (dd, J = 13.7, 5.9 Hz, 2H), 8.16 (d, J = 7.8 Hz, 1H), 7.58 (s, 1H), 7.39-7.30 (m, 2H), 7.15 (d, J = 8.6 Hz, 1H), 5.95 (s, 1H), 4.44 .40 (m, 1H), 3.84 (s, 3H), 3.59 (t, J = 6.6 Hz, 2H), 3.5 .35 (m, 4H), 2.99 (d, J = 12.4 Hz, 6H), 2.68 (t, J = 6.6 Hz, 2H), 2.55 (s, 2H), 2.31 (t, J = 6.7 Hz, 2H), 2.19-2.03 (m, 6H), 1.72-1.58 (m, 10H). Example 13 [00604] N-(2-(1-(2-(2-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl)-2, 7-diazaspiro [3.5] non-7-yl) ethyl) piperidin-4-yl)-6-(2-hydroxyprop- 2-yl)-2H-indazol-5-yl)-6-(trifluoro- methyl) pyridinecarboxamide LC-MS: (ESI, m/z): [M + H] .sup.+ = 846.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.34 (s, 1H), 8.71 (s, 1H), 8.45 (d, J = 7.8 Hz, 1H), 8.37 (dd, J = 14.3, 6.4 Hz, 2H), 8.16 (d, J = 7.8 Hz, 1H), 7.65 (dd, J = 8.6, 2.2 Hz, 1H), 7.60- 7.56 (m, 2H), 7.16 (d, J = 8.8 Hz, 1H), 5.95 (s, 1H), 4.45- 4.42 (m, 1H), 4.03 (s, 2H), 3.86 (s, 3H), 3.72 (s, 2H), 3.59 (t, J = 6.7 Hz, 2H), 3.01 (d, J = 10.1 Hz, 2H), 2.69 (t, J = 6.5 Hz, 2H), 2.49- 2.22 (m, 8H), 2.16- 2.07 (m, 6H), 1.75- 1.68 (m, 4H), 1.62 (s, 6H).

Example 14: N-(2-(1-(2-(4-(3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoyl) piperazin-1-yl) ethyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1810] ##STR00605##

[1811] A mixture of N-(6-methoxy-2-(1-(2-(piperazin-1-yl) ethyl) piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (120 mg, 0.226 mmol), pentafluorophenyl 3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl)-4-methoxybenzoate (49 mg, 0.113 mmol) and N,N-diisopropylethylamine (291 mg, 2.26 mmol) in dimethyl sulfoxide (3 mL) was stirred for 2h at room temperature. The reaction solution was poured into water (50 mL) and stirred for 0.5h, the solution was extracted with ethyl acetate (3-50 mL), and the organic phase was collected, washed with water (2×100 mL) and saturated saline (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The concentrate was purified by preparative HPLC to obtain 28.57 mg of target product as a yellow solid.

[1812] LC-MS: (ESI, m/z): [M+H].sup.+=778.2

[1813] .sup.1H NMR (400 MHz, CDCl3) δ 10.71 (s, 1H), 8.82 (s, 1H), 8.50 (d, J=7.9 Hz, 1H), 8.12 (t, J=7.8 Hz, 1H), 7.93-7.81 (m, 2H), 7.71 (s, 1H), 7.50-7.33 (m, 2H), 7.14-6.93 (m, 2H), 4.52-4.42 (m, 1H), 4.03 (s, 3H), 3.90 (s, 3H), 3.82-3.53 (m, 6H), 3.33 (d, J=11.7 Hz, 2H), 2.82 (t, J=6.6 Hz, 4H), 2.72 (t, J=6.2 Hz, 2H), 2.55-2.43 (m, 6H), 2.35-2.28 (m, 4H).

[1814] The following examples were prepared with reference to the method of example 14:

TABLE-US-00022 Example Structure Name LC-MS/NMR Example 15 [00606] N-(2-(1-(2-(7-(3- (2,4-dioxo- tetrahydropyrim- idin-1 (2H)-yl)-4-methoxy- benzoyl)-2, 7-diazaspiro [3.5] non-2-yl) ethyl) piperidin-4-yl)-6- methoxy-2H-in- dazol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide LC-MS: (ESI, m/z): [M + H] .sup.+ = 818.2 1H NMR (400 MHz, MeOH-d.sub.4) δ 8.76 (d, J = 3.0 Hz, 1H), 8.48- 8.46 (m, 1H), 8.31-8.30 (m, 1H), 8.20 (d, J = 3.0 Hz, 1H), 8.05 (dd, J = 7.8, 2.8 Hz, 1H), 7.42 (dd, J = 22.3, 5.3 Hz, 2H), 7.20 (d, J = 8.6 Hz, 1H), 7.07 (d, J = 3.1 Hz, 1H), 4.45-4.42 (m, 1H), 4.06 (s, 3H), 3.93 (s, 3H), 3.72-3.47 (m, 10H), 3.17- 2.96 (m, 4H), 2.80 (t, J = 6.7 Hz, 2H), 2.58 (d, J = 5.6 Hz, 2H), 2.33-2.21 (m, 6H), 1.89- 1.86 (m, 4H). Example 16 [00607] N-(2-(1-(2-(9-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) ethyl) piperidin-4-yl)-6- methoxy-2H-in- dazol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide LC-MS: (ESI, m/z): [M + H].sup.+ = 850.3 .sup.1H NMR (400 MHz, MeOH-d.sub.4) δ 8.77 (s, 1H), 8.48 (d, J = 7.8 Hz, 1H), 8.31 (t, J = 7.9 Hz, 1H), 8.21 (s, 1H), 8.05 (dd, J = 7.8, 0.8 Hz, 1H), 7.65 (d, J = 8.3 Hz, 1H), 7.54 (d, J = 2.0 Hz, 1H), 7.43 (dd, J = 8.3, 2.0 Hz, 1H), 7.08 (s, 1H), 4.42 (d, J = 7.9 Hz, 1H), 4.06 (s, 3H), 3.87- 3.68 (m, 4H), 3.47 (dd, J = 8.2, 6.5 Hz, 2H), 3.19-3.10 (m, 2H), 2.92-2.80 (m, 2H), 2.62 (d, J = 20.5 Hz, 8H), 2.38-2.27 (m, 2H), 2.26-2.13 (m, 4H), 1.93 (s, 1H), 1.58 (d, J = 57.1 Hz, 8H). Example 17 [00608] N-(2-(1-(2-(9-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) ethyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 878.3 .sup.1H NMR (400 MHz, DMSO) δ 12.37 (s, 1H), 10.51 (s, 1H), 8.71 (s, 1H), 8.45 (d, J = 7.8 Hz, 1H), 8.37 (dd, J = 14.2, 6.3 Hz, 2H), 8.16 (d, J = 7.8 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.59- 7.54 (m, 2H), 7.39 (dd, J = 8.2, 1.9 Hz, 1H), 5.95 (s, 1H), 4.44- 4.40 (m, 1H), 3.81-3.51 (m, 4H), 3.01 (d, J = 9.6 Hz, 2H), 2.74 (dd, J = 8.9, 5.6 Hz, 2H), 2.49-2.00 (m, 16H), 1.62 (s, 6H), 50-1.38 (m, 8H). Example 18 [00609] N-(2-(1-(3-(1-(3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-me- thoxybenzoyl) piperidin-4-yl) propyl) piperidin-4-yl)-6- methoxy-2H-in- dazol-5-yl)- 6-(trifluorometh- yl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 791.6. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.50 (s, 1H), 10.34 (s, 1H), 8.69 (s, 1H), 8.49-8.34 (m, 3H), 8.22 (d, J = 8.5 Hz, 1H), 7.37 (dd, J = 8.4, 2.1 Hz, 1H), 7.32 (d, J = 2.1 Hz, 1H), 7.15 (d, J = 8.1 Hz, 2H), 4.40-4.37 (s, 1H), 3.98 (s, 3H), 3.84 (s, 3H), 3.60 (t, J = 6.6 Hz, 2H), 3.36-3.32 (m, 2H), 2.76 (dd, J = 66.7, 60.1 Hz, 6H), 2.34-2.30 (m, 2H), 2.08 (d, J = 5.4 Hz, 6H), 1.71-1.65 (m, 2H), 1.50- 1.48 (m, 3H), 1.26 (dd, J = 14.5, 6.7 Hz, 2H), 1.08 (dd, J = 20.4, 11.8 Hz, 2H). Example 19 [00610] N-(2-(1-(3-(4-(3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-me- thoxybenzoyl) piperazin-1-yl) propyl) piperidin-4-yl)-6- methoxy-2H-in- dazol-5-yl)- 6-(trifluorometh- yl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 792.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.51 (s, 1H), 10.35 (s, 1H), 8.69 (s, 1H), 8.51-8.33 (m, 3H), 8.20 (d, J = 3.2 Hz, 1H), 7.44-7.30 (m, 2H), 7.17 (t, J = 4.2 Hz, 2H), 4.42-4.35 (m, 1H), 3.99 (s, 3H), 3.85 (s, 3H), 3.58 (dd, J = 25.2, 18.6 Hz, 6H), 3.01 (d, J = 7.1 Hz, 2H), 2.69 (t, J = 6.5 Hz, 2H), 2.36 (dd, J = 15.1, 7.5 Hz, 8H), 2.11 (dd, J = 13.0, 8.5 Hz, 6H), 1.72-1.56 (m, 2H) Example 20 [00611] N-(2-(1-(2-(1-(3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-y)-4-me- thoxybenzoyl) piperidin-4-yl) oxy) ethyl) piperidin-4-yl)-6- methoxy-2H-in- dazol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 793.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.51 (s, 1H), 10.34 (s, 1H), 8.69 (s, 1H), 8.48-8.35 (m, 3H), 8.22 (d, J = 7.6 Hz, 2H), 7.41-7.34 (m, 2H), 7.17-6.95 (m, 2H), 4.42-4.34 (m, 1H), 3.98 (s, 3H), 3.85 (s, 3H), 3.60 (dd, J = 11.4, 5.8 Hz, 7H), 3.28- 3.21 (m, 2H), 3.03 (d, J = 11.2 Hz, 2H), 2.68 (t, J = 6.5 Hz, 2H), 2.57 (t, J = 5.7 Hz, 2H), 2.24 (dd, J = 16.2, 9.0 Hz, 2H), 2.09 (t, J = 10.6 Hz, 4H), 1.87- 1.83 (m, 2H), 1.48-1.45 (m, 2H). Example 21 [00612] N-(2-(1-(2-(3- (3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-me- thoxybenzoyl)-3- azaspiro [5.5] undec-9-yl) ethyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 873.1 .sup.1H NMR (400 MHz, MeOD-d.sub.4) δ 8.72 (s, 1H), 8.47 (d, J = 7.8 Hz, 1H), 8.34-8.23 (m, 2H), 8.02 (d, J = 7.8 Hz, 1H), 7.67 (s, 1H), 7.44 (dd, J = 8.5, 2.1 Hz, 1H), 7.38 (d, J = 2.1 Hz, 1H), 7.18 (d, J = 8.6 Hz, 1H), 4.49 (s, 1H), 3.92 (s, 3H), 3.74-3.70 (m, 4H), 3.50-3.46 (m, 2H), 3.16-3.12 (m, 2H), 2.80 (t, J = 6.7 Hz, 2H), 2.55-2.44 (m, 2H), 2.25 (d, J = 6.1 Hz, 6H), 1.83-1.69 (m, 8H), 1.65-1.61 (m, 3H), 1.42 (s, 6H), 1.21- 1.17 (m, 4H). Example 22 [00613] N-(2-(1-(3-(1-(3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-me- thoxybenzoyl) piperidin-4-yl) propyl) piperidin-4-yl)- 6-(2-hydroxyprop- 2-yl)-2H-inda- zol-5-yl)- 6-(trifluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 819.2. .sup.1H NMR (400 MHz, CDCl.sub.3) δ 12.28 (s, 1H), 8.87 (s, 1H), 8.51 (d, J = 7.8 Hz, 1H), 8.11 (t, J = 7.8 Hz, 1H), 7.95 (s, 1H), 7.84 (d, J = 7.2 Hz, 1H), 7.71 (s, 1H), 7.47 (s, 1H), 7.42 (dd, J = 8.5, 2.1 Hz, 1H), 7.37 (d, J = 2.1 Hz, 1H), 7.00 (d, J = 8.5 Hz, 1H), 4.57 (s, 1H), 3.90 (s, 3H), 3.73- 3.69 (m, 2H), 3.35-3.31 (m, 2H), 2.87-2.33 (m, 12H), 1.80 (s, 6H), 1.77-1.73 (m, 5H), 1.58- 1.56 (m, 2H), 1.33 (dd, J = 15.3, 7.2 Hz, 2H), 1.20-1.17 (m, 2H). Example 23 [00614] N-(2-(1-(2-((1- (3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-me- thoxybenzoyl) piperidin-4-yl) oxy) ethyl) piperdin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-inda- zol-5-yl)-6-(triflu- oromethyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 821.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.33 (s, 1H), 8.72 (s, 1H), 8.45 (d, J = 7.7 Hz, 1H), 8.40 (s, 1H), 8.37 (t, J = 7.8 Hz, 1H), 8.26 (s, 1H), 8.16 (dd, J = 7.8, 0.8 Hz, 1H), 7.58 (s, 1H), 7.39 (dd, J = 8.4, 2.1 Hz, 1H), 7.35 (d, J = 2.1 Hz, 1H), 7.16 (d, J = 8.6 Hz, 1H), 5.94 (s, 1H), 4.48-4.38 (m, 1H), 3.84 (s, 3H), 3.60 (dd, J = 8.7, 4.5 Hz, 7H), 3.28-3.22 (m, 2H), 3.03 (d, J = 11.4 Hz, 2H), 2.68 (t, J = 6.6 Hz, 2H), 2.56 (dd, J = 10.6, 4.7 Hz, 2H), 2.23 (dd, J = 16.3, 9.4 Hz, 2H), 2.10 (t, J = 9.0 Hz, 4H), 1.85 (s, 2H), 1.62 (s, 6H), 1.46 (d, J = 8.7 Hz, 2H). Example 24 [00615] N-(2-(1-(2-(3-((3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-ethoxy- benzoyl)-3- azaspiro[5.5] undec-9-yl) ethyl) piperdin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 887.1. 1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.34 (s, 1H), 8.72 (s, 1H), 8.48-8.29 (m, 3H), 8.16 (d, J = 7.6 Hz, 1H), 7.58 (s, 1H), 7.38-7.28 (m, 2H), 7.13 (d, J = 8.5 Hz, 1H), 5.95 (s, 1H), 4.45-4.40 (m, 1H), 4.11 (q, J = 6.7 Hz, 2H), 3.65-3.40 (m, 6H), 2.99-2.96 (m, 2H), 2.69-2.65 (m, 2H), 2.37-2.33 (m, 2H), 2.11-2.08 (m, 6H), 1.71-1.68 (m, 2H), 1.64-1.59 (m, 6H), 1.52-1.48 (m, 4H), 1.36-1.32 (m, 8H), 1.11-1.08 (m, 4H). Example 25 [00616] N-(2-(1-(2-(7-((3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-ethoxy- benzoyl)-7- azaspiro[3.5] non-2-yl) ethyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 859.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.34 (s, 1H), 8.72 (s, 1H), 8.46-8.35 (m, 3H), 8.16 (d, J = 7.6 Hz, 1H), 7.58 (s, 1H), 7.36-7.27 (m, 2H), 7.13 (d, J = 8.4 Hz, 1H), 5.97-5.94 (m, 1H), 4.50-4.45 (m, 1H), 4.11 (q, J = 7.1 Hz, 2H), 3.62-3.58 (m, 2H), 3.38- 3.33 (m, 6H), 2.99-2.97 (m, 2H), 2.69-2.67 (m, 2H), 2.30- 2.21 (m, 3H), 2.15-2.05 (m, 4H), 2.00-1.95 (m, 2H), 1.64- 1.39 (m, 14H), 1.33 (t, J = 6.9 Hz, 3H). Example 26 [00617] N-(2-(1-(2-(7- ((3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-me- thoxybenzoyl)-7- azaspiro[3.5] non-2-yl) ethyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 845.2. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.33 (s, 1H), 8.72 (s, 1H), 8.40 (dt, J = 21.5, 7.8 Hz, 3H), 8.16 (d, J = 7.2 Hz, 1H), 7.57 (s, 1H), 7.36 (dd, J = 8.4, 2.0 Hz, 1H), 7.32 (d, J = 2.0 Hz, 1H), 7.15 (d, J = 8.6 Hz, 1H), 5.95 (s, 1H), 4.45-4.43 (m, 1H), 3.84 (s, 3H), 3.59 (t, J = 6.6 Hz, 2H), 3.41-3.48 (m, 2H), 3.00-2.98 (m, 2H), 2.68 (t, J = 6.5 Hz, 2H), 2.25-2.23 (m3, 3H), 2.08 (d, J = 4.7 Hz, 6H), 1.97 (t, J = 9.6 Hz, 2H), 1.63- 1.58 (m, 10H), 1.43 (dd, J = 21.6, 12.8 Hz, 4H). Example 27 [00618] N-(2-(1-(2-7-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-7- azospiro [3.5] non-2-yl) ethyl) piperdin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M+ H].sup.+ = 849.1. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.51 (s, 1H), 8.71 (s, 1H), 8.46-8.35 (m, 3H), 8.16 (d, J = 7.7 Hz, 1H), 7.63 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 11.5 Hz, 2H), 7.38 (d, J = 6.4 Hz, 1H), 5.96-5.92 (m, 1H), 4.45-4.42 (m, 1H), 3.69 (d, J = 45.0 Hz, 4H), 3.19-3.15 (m, 1H), 3.00-2.96 (m, 2H), 2.80-2.69 (m, 2H), 2.26-2.23 (m, 3H), 2.08 (d, J = 5.9 Hz, 6H), 1.99-1.95 (m, 2H), 1.52 (d, J = 78.9 Hz, 15H). Example 28 [00619] N-(2-(1-(2-(3-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro [5.5] undec-9-yl) ethyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 877.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.51 (s, 1H), 8.72 (s, 1H), 8.45 (d, J = 7.7 Hz, 1H), 8.37 (dd, J = 14.5, 6.7 Hz, 2H), 8.19-8.14 (m, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.57 (s, 1H), 7.55 (d, J = 2.0 Hz, 1H), 7.42- 7.36 (m, 1H), 5.94 (s, 1H), 4.45- 4.43 (m, 1H), 3.75-3.73 (m, 1H), 3.62 (d, J = 13.1 Hz, 3H), 3.30-3.23 (m, 2H), 3.01-2.99 (m, 2H), 2.74 (dd, J = 9.1, 5.2 Hz, 2H), 2.39-2.37 (m, 2H), 2.09 (d, J = 13.5 Hz, 6H), 1.70 (d, J = 8.8 Hz, 2H), 1.62 (s, 6H), 1.55-1.51 (m, 3H), 1.41-1.37 (m, 4H), 1.28-1.26 (m, 2H), 1.11-1.07 (m, 4H). Example 29 [00620] N-(2-(1-(3-(1-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1)(2H)- yl) benzoyl) piperidin-4-yl) propyl) piperdin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 823.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.51 (s, 1H), 8.72 (s, 1H), 8.40 (dt, J = 22.4, 7.8 Hz, 3H), 8.16 (d, J = 7.8 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.60-7.52 (m, 2H), 7.39 (dd, J = 8.2, 2.0 Hz, 1H), 5.95 (s, 1H), 4.49-4.44 (m, 2H), 3.74-3.71 (m, 1H), 3.68-3.50 (m, 2H), 3.05-2.95 (m, 3H), 2.78-2.70 (m, 3H), 2.38-2.28 (m, 2H), 2.09 (d, J = 5.5 Hz, 6H), 1.83-1.72 (m, 1H), 1.70-1.60 (m, 7H), 1.59 1.43 (m, 3H), 1.30-1.20 (m, 2H), 1.15-1.03 (m, 2H). Example 30 [00621] N-(2-(1-((3-(3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-me- thoxybenzoyl)-3- azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6- methoxy-2H- indazol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 831.1. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.50 (s, 1H), 10.33 (s, 1H), 8.69 (s, 1H), 8.46 (d, J = 7.7 Hz, 1H), 8.41 (t, J = 7.8 Hz, 1H), 8.37 (s, 1H), 8.22 (d, J = 8.6 Hz, 1H), 7.37 (dd, J = 8.5, 2.1 Hz, 1H), 7.33 (d, J = 2.1 Hz, 1H), 7.15 (t, J = 4.3 Hz, 2H), 4.40- 4.36 (m, 1H), 3.98 (s, 3H), 3.84 (s, 3H), 3.60 (t, J = 6.6 Hz, 2H), 3.59-3.33 (m, 4H), 2.97-2.93 (m, 2H), 2.68 (t, J = 6.4 Hz, 2H), 2.13 (d, J = 45.3 Hz, 8H), 1.80-1.63 (m, 2H), 1.63-1.38 (m, 5H), 1.33-1.29 (m, 2H), 1.14-1.08 (m, 4H). Example 31 [00622] N-(2-(1-((3-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro [5.5] undec-9-yl) methyl) piperdin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 863.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.52 (s, 1H), 8.72 (s, 1H), 8.40 (dt, J = 20.0, 7.8 Hz, 3H), 8.16 (dd, J = 7.8, 0.7 Hz, 1H), 7.76-7.47 (m, 3H), 7.39 (dd, J = 8.2, 1.8 Hz, 1H), 5.95 (s, 1H), 4.42 (s, 1H), δ 76 (dd, J = 16.6, 9.4 Hz, 1H), 3.71-3.50 (m, 3H), 3.31-3.29 (m, 2H), 2.96- 2.92 (m, 2H), 2.82-2.69 (m, 2H), 2.12 (d, J = 37.9 Hz, 8H), 1.77-1.66 (m, 2H), 1.66-1.40 (m, 11H), 1.32 (d, J = 30.3 Hz, 2H), 1.11-1.07 (m, 4H). Example 32 [00623] N-(2-(1-(3-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro[5.5] undec-9-yl) methyl) piperdin-4-yl)-6- methoxy-2H- indazol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 835.0 .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.51 (d, J = 2.5 Hz, 2H), 8.69 (s, 1H), 8.46 (d, J = 7.5 Hz, 1H), 8.41 (t, J = 7.8 Hz, 1H), 8.37 (s, 1H), 8.22 (d, J = 7.7 Hz, 1H), 7.64 (d, J = 8.3 Hz, 1H), 7.55 (d, J = 1.9 Hz, 1H), 7.39 (dd, J = 8.3, 1.9 Hz, 1H), 7.16 (s, 1H), 4.40-4.36 (m, 1H), 3.98 (s, 3H), 3.83-3.68 (m, 1H), 3.69- 3.46 (m, 3H), 3.30-3.20 (m, 2H), 2.98-2.95 (m, 2H), 2.81- 2.70 (m, 2H), 2.13 (d, J = 42.7 Hz, 8H), 1.76-1.65 (m, 2H), 1.47-1.42 (m, 7H), 1.12-1.07 (m, 4H). Example 33 [00624] N-(2-(1-((3-(3- (2- 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-me- thoxybenzoyl)-3- azaspiro [5.5] undec-9-yl) methyl) piperdin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 859.1 .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.33 (s, 1H), 8.71 (s, 1H), 8.45 (d, J = 7.7 Hz, 1H), 8.40 (s, 1H), 8.36 (t, J = 7.9 Hz, 1H), 8.16 (d, J = 7.8 Hz, 1H), 7.57 (s, 1H), 7.37 (dd, J = 8.5, 2.0 Hz, 1H), 7.33 (d, J = 2.0 Hz, 1H), 7.15 (d, J = 8.6 Hz, 1H), 5.96-5.93 (m, 1H), 4.45- 4.41 (m, 1H), 3.84 (s, 3H), 3.60 (t, J = 6.6 Hz, 2H), 3.51-3.33 (m, 4H), 2.97-2.95 (m, 2H), 2.68 (t, J = 6.5 Hz, 2H), 2.26- 1.98 (m, 8H), 1.75-1.67 (m, 2H), 1.61-1.44 (m, 11H), 1.33- 1.28 (m, 2H), 1.15-1.07 (m, 4H) Example 34 [00625] N-(2-(1-(2-(3-(3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-fluoro- benzoyl)-3- azaspiro[5.5]undec- 9-yl) ethyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z) : [M + H].sup.+ = 861.1. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.53 (s, 1H), 8.71 (s, 1H), 8.48-8.32 (m, 3H), 8.16 (d, J = 8.0 Hz, 1H), 7.57 (s, 1H), 7.51-7.32 (m, 3H), 5.94 (s, 1H), 4.45-4.35 (m, 1H), 3.75 (t, J = 6.4 Hz, 2H), 3.65-3.47 (m, 2H), 3.05- 2.92 (m, 2H), 2.73 (t, J = 6.6 Hz, 2H), 2.37-2.27 (m, 2H), 2.08 (s, 6H), 1.82-1.20 (m, 19H), 1.16-1.07 (m, 4H). Example 35 [00626] N-(2-(1-(2-(1-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin- 1)(2H)-yl) benzoyl) piperidin-4-yl) ethyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 809.4. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.51 (s, 1H), 8.72 (s, 1H), 8.45 (d, J = 7.9 Hz, 1H), 8.40 (s, 1H), 8.37 (t, J = 7.9 Hz, 1H), 8.16 (d, J = 7.8 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.58 (s, 1H), 7.55 (d, J = 1.9 Hz, 1H), 7.39 (dd, J = 8.2, 1.9 Hz, 1H), 5.95 (s, 1H), 4.50-4.35 (m, 2H), 3.80-3.72 (m, 1H), 3.66-3.51 (m, 2H), 3.11-2.94 (m, 3H), 2.84-2.66 (m, 3H), 2.42-2.35 (m, 2H), 2.09 (s, 6H), 1.85-1.73 (m, 1H), 1.71- 1.53 (m, 8H), 1.49-1.39 (m, 2H), 1.20-1.05 (m, 2H). Example 36 [00627] N-(2-(1-((7-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-7- azaspiro [3.5] non-2-yl) methyl) piperdin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 835.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.51 (s, 1H), 8.71 (s, 1H), 8.45 (d, J = 7.8 Hz, 1H), 8.40 (s, 1H), 8.36 (t, J = 7.9 Hz, 1H), 8.16 (d, J = 7.8 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.57 (s, 1H), 7.55 (d, J = 1.6 Hz, 1H), 7.38 (dd, J = 8.2, 1.8 Hz, 1H), 5.94 (s, 1H), 4.48-4.35 (m, 1H), 3.82-3.70 (m, 1H), 3.66-3.41 (m, 3H), 3.30-3.11 (m, 2H), 2.98-2.86 (m, 2H), 2.80-2.70 (m, 2H), 2.44-2.38 (m, 2H), 2.20-2.02 (m, 6H), 2.02-1.97 (m, 2H), 1.70-1.40 (m, 13H) Example 37 [00628] N-(2-(1-(2-(3-(3- (3-, 2, 4-dioxotetrahydro- pyrimidin-1)(2H)- yl)-4-(trifluoro- methoxy) benzoyl)-3- azaspiro[5.5] undec-9-yl) ethyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2 H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 927.4 .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.55 (s, 1H), 8.72 (s, 1H), 8.45 (d, J = 7.9 Hz, 1H), 8.40 (s, 1H), 8.36 (t, J = 8.0 Hz, 1H), 8.16 (d, J = 7.7 Hz, 1H), 7.60 (s, 1H), 7.57 (s, 1H), 7.50 (dd, J = 20.5, 8.2 Hz, 2H), 5.94 (s, 1H), 4.50-4.36 (m, 1H), 3.80-3.53 (m, 4H), 3.31- 3.19 (m, 2H), 3.04-2.94 (m, 2H), 2.74-2.66 (m, 2H), 2.40- 2.30 (m, 2H), 2.08 (s, 6H), 1.77- 1.19 (m, 17H), 1.18-1.02 (m, 4H). Example 38 [00629] N-(2-(1-(2-(3-(3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-(trifluoro- methyl) benzoyl)-3- azaspiro[5.5] undec-9-yl) ethyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 911.0. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.54 (s, 1H), 8.71 (s, 1H), 8.45 (d, J = 7.7 Hz, 1H), 8.40 (s, 1H), 8.39-8.33 (m, 1H), 8.16 (d, J = 7.8 Hz, 1H), 7.87 (d, J = 8.2 Hz, 1H), 7.71 (s, 1H), 7.59 (d, J = 10.8 Hz, 1H), 7.51 (s, 1H), 5.95 (s, 1H), 4.50-4.35 (m, 1H), 3.97- 3.80 (m, 1H), 3.65-3.55 (m, 2H), 3.55-3.41 (m, 2H), 3.27- 3.20 (m, 2H), 3.02-2.94 (m, 2H), 2.69 (dd, J = 10.1, 5.7 Hz, 2H), 2.38-2.30 (m, 2H), 2.13- 2.03 (m, 6H), 1.70 (d, J = 10.3 Hz, 2H), 1.62 (s, 6H), 1.57- 1.49 (m, 3H), 1.41 (d, J = 26.9 Hz, 4H), 1.31-1.23 (m, 2H), 1.17-1.10 (m, 4H). Example 39 [00630] N-(2-(1-((7-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-7- azospiro [3.5] non-2-yl) methyl) piperidin-4-yl)-6- methoxy-2H- indazol-5-yl)-6- (trifluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 807.0 .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.51 (s, 1H), 10.50 (s, 1H), 8.68 (s, 1H), 8.46 (d, J = 7.8 Hz, 1H), 8.40 (t, J = 7.8 Hz, 1H), 8.36 (s, 1H), 8.22 (d, J = 7.3 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.55 (s, 1H), 7.38 (d, J = 8.4 Hz, 1H), 7.16 (s, 1H), 4.40-4.30 (m, 1H), 3.98 (s, 3H), 3.80- 3.70 (m, 1H), 3.68-3.36 (m, 4H), 3.22-3.13 (m, 1H), 2.93 (d, J = 8.3 Hz, 2H), 2.81-2.67 (m, 2H), 2.45-2.37 (m, 3H), 2.20-1.86 (m, 8H), 1.70-1.40 (m, 6H). Example 40 [00631] N-(2-(1-(2-(3- (3-(2, 6-dioxopiperidin- 3-yl)-4-fluoro- benzoyl)-3-azaspiro [5.5] undec-9-yl) ethyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 860.3. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.92 (s, 1H), 8.72 (s, 1H), 8.45 (d, J = 7.6 Hz, 1H), 8.40 (s, 1H), 8.37 (t, J = 7.8 Hz, 1H), 8.16 (d, J = 7.7 Hz, 1H), 7.57 (s, 1H), 7.36 (d, J = 6.8 Hz, 2H), 7.26 (t, J = 9.6 Hz, 1H), 5.95 (s, 1H), 4.44 (s, 1H), 4.12 (dd, J = 12.6, 4.7 Hz, 1H), 3.56 (s, 2H), 3.00 (s, 2H), 2.84- 2.64 (m, 1H), 2.57 (s, 1H), 2.38 (s, 2H), 2.30-1.92 (m, 9H), 1.70 (d, J = 7.5 Hz, 2H), 1.62 (s, 6H), 1.57-1.20 (m, 10H), 1.09 (s, 4H). Example 41 [00632] N-(2-(1-((3-(3- (2, 6-dioxopiperidin- 3-yl)-4-fluoro- benzoyl)-3-azaspiro [5.5] undec-9-yl) methyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-inda- zol-5-yl)-6-(triflu- oromethyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 846.1 .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.37 (s, 1H), 10.92 (s, 1H), 8.72 (s, 1H), 8.45 (d, J = 7.8 Hz, 1H), 8.40 (s, 1H), 8.37 (t, J = 7.8 Hz, 1H), 8.17 (d, J = 7.9 Hz, 1H), 7.58 (s, 1H), 7.37 (d, J = 6.9 Hz, 2H), 7.27 (t, J = 9.6 Hz, 1H), 5.95 (s, 1H), 4.47-4.38 (m, 1H), 4.13 (dd, J = 12.7, 4.9 Hz, 1H), 3.67-3.50 (m, 2H), 3.29-3.20 (m, 1H), 3.02-2.87 (m, 2H), 2.84-2.66 (m, 1H), 2.60-2.55 (m, 1H), 2.32-1.97 (m, 10H), 1.72 (d, J = 9.1 Hz, 2H), 1.67-1.21 (m, 14H), 1.18- 0.95 (m, 4H). Example 42 [00633] N-(2-(1-((3-(3- (2, 6-dioxopiperidin- 3-yl)-4-fluoro- benzoyl)-3-azaspiro [5.5] undec-9-yl) methyl) piperidin-4-yl)-6- methoxy-2H- indazol-5-yl)-6- (trifluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 818.1 .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.92 (s, 1H), 10.50 (s, 1H), 8.69 (s, 1H), 8.46 (d, J = 7.8 Hz, 1H), 8.40 (t, J = 7.8 Hz, 1H), 8.37 (s, 1H), 8.22 (d, J = 7.7 Hz, 1H), 7.36 (d, J = 6.9 Hz, 2H), 7.26 (t, J = 9.6 Hz, 1H), 7.16 (s, 1H), 4.42-4.31 (m, 1H), 4.12 (dd, J = 12.7, 4.9 Hz, 1H), 3.98 (s, 3H), 3.66-3.50 (m, 2H), 3.30-3.24 (m, 1H), 2.99-2.88 (m, 2H), 2.82-2.68 (m, 1H), 2.55 (d, J = 17.2 Hz, 1H), 2.33- 1.96 (m, 10H), 1.76-1.66 (m, 2H), 1.64-1.20 (m, 8H), 1.19- 0.96 (m, 4H). Example 43 [00634] N-(2-(1-(2 (7-(4-chloro- 3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-7- azospiro [3.5] non-2-yl) ethyl) piperidin-4-yl)-6- methoxy-2H- indazol-5-yl)-6- (trifluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 821.1. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.50 (s, 2H), 8.69 (s, 1H), 8.46 (d, J = 7.3 Hz, 1H), 8.41 (t, J = 7.8 Hz, 1H), 8.37 (s, 1H), 8.22 (d, J = 8.6 Hz, 1H), 7.63 (d, J = 8.2 Hz, 1H), 7.54 (s, 1H), 7.38 (d, J = 8.1 Hz, 1H), 7.16 (s, 1H), 4.42-4.30 (m, 1H), 3.98 (s, 3H), 3.80-3.70 (m, 1H), 3.65-3.45 (m, 2H), 3.27-3.10 (m, 3H), 3.02-2.90 (m, 2H), 2.74 (dd, J = 11.0, 6.7 Hz, 2H), 2.30-2.16 (m, 3H), 2.13-1.90 (m, 8H), 1.66-1.35 (m, 8H). Example 44 [00635] N-(2-(1-((3-(3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-fluoro- benzoyl)-3- azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6- methoxy-2H- indazol-5-yl)-6- (trifluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 819.0 .sup.1HNMR (400 MHz, DMSO) δ 10.52 (d, J = 11.5 Hz, 2H), 8.69 (s, 1H), 8.49-8.33 (m, 3H), 8.22 (d, J = 7.6 Hz, 1H), 7.51 (d, J = 7.0 Hz, 1H), 7.42-7.30 (m, 2H), 7.16 (s, 1H), 4.39-4.35 (m, 1H), 3.98 (s, 3H), 3.76 (t, J = 6.5 Hz, 2H), 3.61-3.55 (m, 2H), 3.32-3.26 (m, 2H), 2.97-2.90 (m, 2H), 2.73 (t, J = 6.6 Hz, 2H), 2.19-2.13 (m, 2H), 2.07 (d, J = 5.5 Hz, 6H), 1.71 (d, J = 9.2 Hz, 2H), 1.64-1.43 (m, 5H), 1.40-1.01 (m, 6H). Example 45 [00636] N-(2-(1-((3-(3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-fluoro- benzoyl)-3- azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6- (2-hydroxyprop- 2-yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 847.6 .sup.1HNMR(400 MHz, DMSO) δ 12.37 (s, 1H), 10.55 (s, 1H), 8.72 (s, 1H), 8.45 (d, J = 7.9 Hz, 1H), 8.40 (s, 1H), 8.36 (t, J = 8.0 Hz, 1H), 8.16 (d, J = 7.7 Hz, 1H), 7.60 (s, 1H), 7.57 (s, 1H), 7.50 (dd, J = 20.5, 8.2 Hz, 2H), 5.94 (s, 1H), 4.50-4.36 (m, 1H), 3.85 (t, J = 8.0 Hz, 1H), 3.66-3.53 (m, 2H), 3.31-3.19 (m, 2H), 3.05-2.96 (m, 2H), 2.75 (t, J = 8.0 Hz, 1H), 2.25- 2.05 (m, 8H), 1.75-1.61 (m, 2H), 1.58-1.40 (m, 11H), 1.38- 1.19 (m, 2H), 1.18-1.03 (m, 4H). Example 46 [00637] N-(2-(1-(2-(3-(3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl)-4-fluoro- benzoyl)-3- azaspiro[5.5] undec-9-yl) ethyl) piperidin-4-yl)-6- methoxy-2H- indazol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 833.1 .sup.1H NMR (400 MHz, DMSO) δ 10.52 (d, J = 10.5 Hz, 2H), 8.69 (s, 1H), 8.46 (d, J = 7.7 Hz, 1H), 8.40 (dd, J = 15.5, 7.6 Hz, 2H), 8.22 (d, J = 7.6 Hz, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.40-7.33 (m, 2H), 7.16 (s, 1H), 4.42- 3.33 (m, 1H), 3.98 (s, 3H), 3.75 (t, J = 6.6 Hz, 2H), 3.62-3.48 (m, 2H), 3.35-3.24 (m, 1H), 3.04-2.92 (m, 2H), 2.73 (t, J = 6.7 Hz, 2H), 2.41-2.30 (m, 2H), 2.07 (d, J = 5.4 Hz, 6H), 1.70 (d, J = 8.1 Hz, 2H), 1.60- 1.20 (m, 10H), 1.19-1.02 (m, 4H). Example 47 [00638] N-(2-(1-(2-(3- (3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro[5.5] undec-9-yl) ethyl) piperidin-4-yl)-6- methoxy-2H-in- dazol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H].sup.+ = 815.3 .sup.1H NMR (400 MHz, DMSO) δ 10.51 (s, 1H), 10.42 (s, 1H), 8.69 (s, 1H), 8.47 (d, J = 7.6 Hz, 1H), 8.40 (dd, J = 15.0, 7.3 Hz, 2H), 8.23 (d, J = 7.6 Hz, 1H), 7.46 (t, J = 7.7 Hz, 1H), 7.40 (d, J = 8.2 Hz, 1H), 7.36 (s, 1H), 7.23 (d, J = 6.9 Hz, 1H), 7.17 (s, 1H), 4.45-4.32 (m, 1H), 4.02- 3.98 (s, 3H), 3.83 (t, J = 6.7 Hz, 2H), 3.62-3.58 (m, 2H), 3.32- 3.28 (m, 2H), 3.05-2.92 (m, 2H), 2.72 (t, J = 6.6 Hz, 2H), 2.40-2.30 (m, 2H), 2.15-2.02 (m, 5H), 1.71 (d, J = 8.7 Hz, 2H), 1.60-1.22 (m, 10H), 1.15-1.05 (m, 4H). Example 48 [00639] N-(2-(1-(2-(3-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl )-3- azaspiro[5.5] undec-9-yl) ethyl) piperidin-4-yl)-6- methoxy-2H-in- dazol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide (ESI, m/z): [M + H] .sup.+ = 849.4. .sup.1H NMR (400 MHz, DMSO) δ 10.51 (d, J = 2.0 Hz, 2H), 8.69 (s, 1H), 8.46 (d, J = 7.8 Hz, 1H), 8.40 (dd, J = 15.2, 7.3 Hz, 2H), 8.22 (d, J = 7.7 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.55 (s, 1H), 7.39 (d, J = 8.1 Hz, 1H), 7.16 (s, 1H), 4.45-4.30 (m, 1H), 3.98 (s, 3H), 3.81-3.70 (m, 1H), 3.67-3.55 (m, 3H), 3.05-2.95 (m, 2H), 2.79-2.71 (m, 2H), 2.40-2.30 (m, 2H), 2.20-1.95 (m, 6H), 1.75-1.65 (m, 2H), 1.60-1.20 (m, 11H), 1.18-1.02 (m, 4H). Example 49 [00640] N-(2-((1r,4r)-4- (((2-(1-(4-chloro- 3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl) piperidin-4-yl) ethyl) (methyl) amino) methyl) cyclohexyl)-6- methoxy-2H-inda- zol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide (ESI, m/z) [M + H] .sup.+ = 823.4. .sup.1H NMR (400 MHz, DMSO) δ 10.51 (s, 2H), 9.93 (s, 1H), 8.70 (d, J = 4.1 Hz, 1H), 8.50-8.32 (m, 3H), 8.22 (d, J = 7.6 Hz, 1H), 7.65 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.40 (dd, J = 8.2, 2.0 Hz, 1H), 7.15 (d, J = 8.7 Hz, 1H), 4.42 (d, J = 11.3 Hz, 2H), 3.99 (s, 3H), 3.83-3.69 (m, 1H), 3.64 (d, J = 6.3 Hz, 2H), 3.22- 2.89 (m, 5H), 2.75 (t, J = 7.5 Hz, 6H), 2.12 (d, J = 15.8 Hz, 3H), 1.96 (d, J = 10.5 Hz, 4H), 1.84- 1.55 (m, 5H), 1.36-1.07 (m, 4H). Example 50 [00641] N-(2-((1r,4r)-4- ((9-(4-chloro-3- (2, 4-dioxo tetrahydropyrim- idin-1 (2H)-yl) benzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) methyl) cyclohexyl)-6- methoxy-2H-inda- zol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide (ES, m/z): [M + H].sup.+ = 835.4 .sup.1H NMR (400 MHz, DMSO) δ 10.51 (s, 1H), 10.51 (s, 1H), 8.70 (d, J = 3.0 Hz, 1H), 8.46 (d, J = 7.6 Hz, 1H), 8.41 (t, J = 7.8 Hz, 1H), 8.33 (s, 1H), 8.22 (d, J = 7.6 Hz, 1H), 7.66 (d, J = 8.2 Hz, 1H), 7.56 (s, 1H), 7.40 (d, J = 8.4 Hz, 1H), 7.14 (s, 1H), 4.41 (t, J = 11.4 Hz, 1H), 3.99 (s, 3H), 3.77-3.71 (m, 3H), 3.42-3.27 (m, 4H), 3.13-2.93 (m, 4H), 2.75 (t, J = 6.8 Hz, 2H), 2.22-2.13 (m, 2H), 2.02- 1.85 (m, 7H), 1.75-1.55 (m, 5H), 1.50-1.35 (m, 2H), 1.32- 1.23 (m, 2H). Example 51 [00642] N-(2-((1r,4r)-4- ((((1-(4-chloro-3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl) piperdin-4-yl) methyl) (methyl) amino) methyl) cyclohexyl)-6- methoxy-2H-inda- zol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide LC-MS: (ES, m/z): [M + H].sup.+ = 809.3 .sup.1H NMR (400 MHz, DMSO) δ 10.51 (s, 2H), 8.70 (d, J = 3.2 Hz, 1H), 8.46 (d, J = 7.5 Hz, 1H), 8.41 (t, J = 7.8 Hz, 1H), 8.34 (s, 1H), 8.22 (d, J = 7.6 Hz, 1H), 7.66 (dd, J = 8.2, 3.1 Hz, 1H), 7.56 (d, J = 1.7 Hz, 1H), 7.40 (dd, J = 8.2, 2.0 Hz, 1H), 7.15 (d, J = 8.4 Hz, 1H), 4.40 (t, J = 14.0 Hz, 2H), 3.99 (s, 3H), 3.74 (m, 4H), 3.18-3.04 (m, 3H), 3.02-2.92 (m, 2H), 2.87- 2.82 (m, 3H), 2.75 (t, J = 7.0 Hz, 2H), 2.22-2.12 (m, 3H), 2.07-1.89 (m, 5H), 1.87-1.54 (m, 2H), 1.35-1.18 (m, 4H). Example 52 [00643] N-(2-((1r,4r)-4- ((2-(1-(3-(2, 6-dioxopiperidin- 3-yl)-4-fluoro- benzoyl) piperidin-4-yl) ethyl) (methyl) amino) methyl) cyclohexyl)-6- methoxy-2H-inda- zol-5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ES, m/z): [M + H].sup.+ = 806.4 .sup.1H NMR (400 MHz, DMSO) δ 10.90 (s, 1H), 10.50 (s, 1H), 8.69 (s, 1H), 8.46 (d, J = 7.5 Hz, Hi), 8.41 (t, J = 7.8 Hz, 1H), 8.33 (s, 1H), 8.22 (d, J = 7.6 Hz, 1H), 7.36 (d, J = 6.9 Hz, 2H), 7.26 (t, J = 9.7 Hz, 1H), 7.14 (s, 1H), 4.50-4.21 (m, 2H), 4.12 (dd, J = 12.6, 4.9 Hz, 1H), 3.98 (s, 3H), 3.66-3.45 (m, 1H), 3.10-2.95 (m, 2H), 2.79-2.66 (m, 2H), 2.60-2.55 (m, 2H), 2.37-2.21 (m, 3H), 2.20-2.10 (m, 4H), 2.07-1.97 (m, 2H), 1.96-1.85 (m, 4H), 1.70-1.35 (m, 6H), 1.20-1.03 (m, 4H). Example 53 [00644] N-(2-((1r,4r)-4- (((3-(4-chloo-3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro [5.5] undec-9-yl) (methyl) amino) methyl) cyclohexyl)-6- methoxy-2H-inda- zol-5-yl)-6-(triflu- oromethyl) pyridinecarbox- amide LC-MS: (ES, m/z): [M + H].sup.+ = 863.2 .sup.1H NMR (400 MHz, DMSO) δ 10.50 (d, J = 3.3 Hz, 2H), 8.68 (s, 1H), 8.46 (d, J = 7.7 Hz, 1H), 8.40 (t, J = 7.8 Hz, 1H), 8.32 (s, 1H), 8.21 (d, J = 7.7 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 1.5 Hz, 1H), 7.39 (d, J = 7.7 Hz, 1H), 7.15 (s, 1H), 4.39- 4.28 (m, 1H), 3.98 (s, 3H), 3.83- 3.44 (m, 4H), 3.33-3.26 (m, 2H), 2.75 (dd, J = 8.9, 5.1 Hz, 2H), 2.24 (d, J = 36.3 Hz, 6H), 2.13 (d, J = 11.4 Hz, 2H), 1.99- 1.81 (m, 4H), 1.76 (d, J = 12.3 Hz, 2H), 1.57-1.45 (m, 5H), 1.30 (d, J = 30.1 Hz, 4H), 1.12- 1.04 (m, 4H). Example 54 [00645] N-(2-((1r,4r)-4- (2-(9-(4-chloro-3- (2, 4-dioxo- tetrahydropyrim- idin-1 (2H)-yl) benzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) ethyl) cyclohexyl)-6- methoxy-2H-inda- zol-5-yl)-6-(tri- fluoromethyl) pyridinecarbox- amide LC-MS: (ES, m/z): [M + H].sup.+ = 849.4 .sup.1H NMR (400 MHz, DMSO) δ 10.51 (d, J = 5.6 Hz, 2H), 8.68 (s, 1H), 8.46 (d, J = 7.7 Hz, 1H), 8.40 (t, J = 7.8 Hz, 1H), 8.33 (s, 1H), 8.22 (d, J = 8.4 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 1.9 Hz, 1H), 7.39 (dd, J = 8.2, 2.0 Hz, 1H), 7.15 (s, 1H), 4.35 (t, J = 11.8 Hz, 1H), 3.98 (s, 3H), 3.82-3.52 (m, 4H), 3.35-3.21 (m, 1H), 2.74 (dd, J = 8.8, 5.6 Hz, 2H), 2.39-2.27 (m, 6H), 2.11 (d, J = 9.1 Hz, 2H), 1.93-1.79 (m, 4H), 1.44 (d, J = 40.7 Hz, 12H), 1.16 (d, J = 10.6 Hz, 2H) Example 55 [00646] N-(2-((1r,4r)-4- (((2-(1-(4-chloro- 3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl) piperidin-4-yl) ethyl) (methyl) amino) methyl) cyclohexyl)-6-(2- hydroxyprop-2- yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ES, m/z): [M+ H] .sup.+ = 851.5. .sup.1H NMR (400 MHz, DMSO) δ 12.36 (s, 1H), 10.51 (s, 1H), 8.89 (s, 1H), 8.72 (s, 1H), 8.45 (d, J = 7.8 Hz, 1H), 8.40-8.33 (m, 2H), 8.16 (d, J = 7.7 Hz, 1H), 7.66 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 2.3 Hz, 2H), 7.39 (dd, J = 8.2, 1.9 Hz, 1H), 5.94 (s, 1H), 4.49-4.29 (m, 2H), 3.86-3.71 (m, 1H), 3.64 (d, J = 5.4 Hz, 2H), 3.12-2.98 (m, 4H), 2.76 (dd, J = 15.7, 9.4 Hz, 6H), 2.17 (d, J = 10.4 Hz, 2H), 1.97 (d, J = 10.3 Hz, 5H), 1.72- 1.54 (m, 13H), 1.17 (d, J = 14.4 Hz, 4H). Example 56 [00647] N-(2-((1r,4r)-4- ((3-(4-chloro-3- (2, (4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro[5.5] undec-9-yl) (methyl) amino) cyclohexyl)-6- methoxy-2H-inda- zol-5-yl)-6-(triflu- oromethyl) pyridinecarbox- amide LC-MS: (ESI, m/z): [M + H].sup.+ = 849.4. 1H NMR (400 MHz, DMSO) δ 10.51 (s, 2H), 8.68 (s, 1H), 8.53- 8.38 (m, 2H), 8.32 (s, 1H, FA), 8.28-8.16 (m, 2H), 7.65 (d, J = 8.2 Hz, 1H), 7.55 (s, 1H), 7.40 (d, J = 7.3 Hz, 1H), 7.13 (s, 1H), 4.45-4.35 (m, 1H), 3.98 (s, 3H), 3.81-3.70 (m, 1H), 3.68-3.55 (m, 3H), 3.38-3.25 (s, 2H), 3.01-2.96 (m, 1H), 2.76 (dd, J = 8.5, 5.7 Hz, 3H), 2.39 (s, 3H), 2.22-2.14 (m, 2H), 2.10-1.90 (m, 4H), 1.79 (d, J = 12.7 Hz, 2H), 1.72-1.42 (m, 8H), 1.38-1.10 (m, 4H). Example 57 [00648] N-(2-(3-((1-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl) piperidin-4-yl) methyl)-3-azaspiro [5.5] undec-9-yl)-6- methoxy-2H-inda- zol-5-yl)-6-(triflu- oromethyl) pyridinecarbox- amide LC-MS: (ES, m/z): [M + H].sup.+ = 835.2 .sup.1H NMR (400 MHz, DMSO) δ 10.50 (s, 2H), 8.68 (s, 1H), 8.46 (d, J = 7.8 Hz, 1H, 8.40 (t, J = 7.8 Hz, 1H), 8.36 (s, 1H), 8.22 (d, J = 7.8 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.54 (s, 1H), 7.38 (d, J = 8.4 Hz, 1H), 7.17 (s, 1H), 4.42-4.32 (m, 2H), 3.98 (s, 3H), 3.69 (d, J = 46.3 Hz, 3H), 3.10-3.00 (m, 1H), 2.87-2.70 (m, 3H), 2.39-2.28 (m, 4H), 2.16 (d, J = 6.6 Hz, 2H), 2.09- 1.98 (m, 2H), 1.95-1.88 (m, 2H), 1.79 (d, J = 12.1 Hz, 4H), 1.69-1.52 (m, 3H), 1.43-1.33 (m, 2H), 1.29 (t, J = 12.2 Hz, 2H), 1.07 (d, J = 10.2 Hz, 2H). Example 58 [00649] N-(2-((1r,4r)-4- (9-(4-chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) cyclohexyl)-6- methoxy-2H-inda- zol-5-yl)-6-(triflu- oromethyl) pyridinecarbox- amide LC-MS: (ES, m/z): [M + H]+ = 821.4. 1H NMR (400 MHz, DMSO) δ 10.52 (s, 2H), 8.68 (s, 1H), 8.53- 8.35 (m, 2H), 8.35-8.30 (m, 2H, 1FA), 8.21 (d, J = 7.8 Hz, 1H), 7.66 (d, J = 8.3 Hz, 1H), 7.55 (d, J = 1.8 Hz, 1H), 7.41 (dd, J = 8.2, 1.9 Hz, 1H), 7.13 (s, 1H), 4.42-4.38 (m, 1H), 3.98 (s, 3H), 3.82-3.71 (m, 1H), 3.64-3.54 (m, 3H), 3.32 (s, 2H), 2.78 (d, J = 5.3 Hz, 7H), 2.20 (d, J = 10.7 Hz, 2H), 2.06 (d, J = 9.9 Hz, 2H), 2.00-1.86 (m, 2H), 1.65-1.35 (m, 10H). Example 59 [00650] N-(2-((1s,4s)-4- (9-(4-chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3, 9-diazaspiro[5.5] undec-3-yl) cyclohexyl)-6- methoxy-2H-inda- zol-5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ES, m/z): [M + H].sup.+ = 821.3 .sup.1H NMR (400 MHz, DMSO) δ 10.50 (s, 2H), 8.69 (s, 1H), 8.47 (d, J = 7.7 Hz, 1H), 8.40 (t, J = 7.8 Hz, 1H), 8.36 (s, 1H), 8.22 (d, J = 7.7 Hz, 1H), 7.63 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 1.8 Hz, 1H), 7.39 (dd, J = 8.3, 1.9 Hz, 1H), 7.19 (s, 1H), 4.50 (m, 1H), 3.98 (s, 3H), 3.65 (dd, J = 31.3, 24.7 Hz, 4H), 3.29 (m, 2H), 2.74 (dd, J = 8.9, 5.5 Hz, 2H), 2.46- 2.22 (m, 7H), 1.87 (m, 4H), 1.49 (t, J = 40.4 Hz, 10H). Example 60 [00651] N-(2-((1r,4r)-4- ((3-(4-chloro-3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro[5.5] undec-9-yl (ethyl) amino) cyclohexyl)-6- methoxy-2H-inda- zol-5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ESI, m/z): [M + H] .sup.+ = 863.4. 1H NMR (400 MHz, DMSO) δ 10.50 (s, 2H), 8.68 (s, 1H), 8.48- 8.37 (m, 2H), 8.30 (s, 1H), 8.22 (d, J = 7.6 Hz, 1H), 7.64 (d, J = 8.3 Hz, 1H), 7.55 (d, J = 1.7 Hz, 1H), 7.39 (d, J = 8.1 Hz, 1H), 7.14 (s, 1H), 4.40-4.30 (m, 1H), 3.98 (s, 3H), 3.80- 3.70 (m, 1H), 3.62 (d, J = 12.5 Hz, 3H), 3.29 (s, 2H), 2.72 (dd, J = 20.0, 13.1 Hz, 3H), 2.65- 2.50 (m, 2H), 2.18-2.08 (m, 2H), 2.02-1.85 (m, 2H), 1.85- 1.70 (m, 4H), 1.64-1.07 (m, 13H), 0.98 (s, 3H). Example 61 [00652] N-(2-(1-((3-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro [5.5] undec-9-yl) methyl) piperidin-4-yl)-6- (trifluoromethoxy)- 2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ESI, m/z): [M + H].sup.+ = 889.2. .sup.1H NMR (400 MHz, DMSO) δ 10.51 (s, 1H), 10.40 (s, 1H), 8.68 (s, 1H), 8.62 (s, 1H), 8.44 (dt, J = 15.7, 7.7 Hz, 2H), 8.25 (d, J = 7.5 Hz, 1H), 7.81 (s, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 1.8 Hz, 1H), 7.39 (d, J = 10.0 Hz, 1H), 4.50 (s, 1H), 3.75 (s, 1H), 3.60 (s, 3H), 3.30 (s, 2H), 2.96 (s, 2H), 2.81-2.72 (m, 2H), 2.18 (s, 2H), 2.10 (d, J = 5.3 Hz, 6H), 1.70 (s, 2H), 1.60 (s, 2H), 1.51 (s, 3H), 1.36 (s, 1H), 1.31-1.21 (m, 1H), 1.12 (d, J = 19.7 Hz, 4H). Example 62 [00653] N-(2-((1r,4r)-4- ((((1-(4-chloro-3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl) piperidin-4-yl) methyl (methyl) amino) methyl) cyclohexyl)-6-(2- hydroxyprop-2- yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ES, m/z): [M + H] .sup.+ = 837.5. .sup.1H NMR (400 MHz, DMSO) δ 12.35 (s, 1H), 10.50 (s, 1H), 8.71 (s, 1H), 8.45 (d, J = 8.6 Hz, 1H), 8.36 (d, J = 5.7 Hz, 2H), 8.16 (d, J = 7.2 Hz, 1H), 7.64 (d, J = 7.1 Hz, 1H), 7.56 (d, J = 5.3 Hz, 2H), 7.39 (d, J = 7.8 Hz, 1H), 5.93 (s, 1H), 4.41 (m, 2H), 3.68 (d, J = 53.7 Hz, 3H), 3.06 (m, 1H), 2.75 (m, 3H), 2.15 (m, 9H), 1.99-1.87 (m, 4H), 1.79 (m, 2H), 1.62 (m, 8H), 1.08 (m, 4H). Example 63 [00654] N-(2-((1r,4r)-4- (((3-(4-chloro-3- (2, (4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro[5.5] undec-9-yl) (methyl) amino) methyl) cyclohexyl)-6-(2- hydroxyprop-2- yl)-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ES, m/z): [M + H].sup.+ = 891.4. H-NMR-MC21-308-111: .sup.1H NMR (400 MHz, DMSO) δ 12.36 (s, 1H), 10.51 (.sub.S, 1H), 8.71 (s, 1H), 8.45 (d, J = 7.6 Hz, 1H), 8.36 (dd, J = 9.7, 5.9 Hz, 2H), 8.21-8.11 (m, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.59-7.52 (m, 2H), 7.39 (d, J = 8.2 Hz, 1H), 5.94 (s, 1H), 4.41 (m, 1H), 3.76 (d, J = 7.8 Hz, 1H), 3.69- 3.49 (m, 3H), 3.29 (m, 2H), 2.81- 2.70 (m, 2H), 2.24 (d, J = 37.0 Hz, 6H), 2.13 (d, J = 10.7 Hz, 2H), 1.90 (dd, J = 23.9, 11.1 Hz, 4H), 1.77 (d, J = 11.7 Hz, 2H), 1.62 (s, 6H), 1.51 (m, 5H), 1.31 (d, J = 31.9 Hz, 4H), 1.14-1.04 (m, 4H). Example 64 [00655] N-(2-((1r,4r)-4- ((2-(1-(4-chloro- 3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl) piperidin-4-yl) ethoxy) methyl) cyclohexyl)-6- methoxy-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ESI, m/z) [M + H] .sup.+ = 810.3. 1H NMR (400 MHz, DMSO) δ 10.50 (s, 2H), 8.68 (s, 1H), 8.48- 8.37 (m, 2H), 8.33 (s, 1H), 8.22 (d, J = 7.1 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 1.9 Hz, 1H), 7.38 (dd, J = 8.2, 2.0 Hz, 1H), 7.16 (s, 1H), 4.50- 4.32 (m, 2H), 3.98 (s, 3H), 3.75 (s, 1H), 3.63 (d, J = 6.5 Hz, 2H), 3.43 (t, J = 6.4 Hz, 2H), 3.25 (d, J = 6.2 Hz, 2H), 3.10-3.01 (m, 1H), 2.83-2.68 (m, 3H), 2.14 (d, J = 9.1 Hz, 2H), 1.89 (d, J = 10.7 Hz, 4H), 1.80-1.58 (m, 4H), 1.49 (d, J = 6.3 Hz, 2H), 1.25-1.07 (m, 4H). Example 65 [00656] N-(2-((1r,4r)-4- (((1′-(4-chloro-3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-[1,4′-bi- piperidin]-4-yl) (methyl) amino) methyl) cyclohexyl)-6- methoxy-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ESI, m/z): [M + H] .sup.+ = 878.3 .sup.1H NMR (400 MHz, DMSO) δ 10.50 (s, 2H), 8.68 (s, 1H), 8.46 (d, J = 7.5 Hz, 1H), 8.40 (t, J = 7.9 Hz, 1H), 8.33 (s, 1H), 8.22 (d, J = 8.5 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.40 (dd, J = 8.2, 2.0 Hz, 1H), 7.15 (s, 1H), 4.38 (dd, J = 33.5, 23.2 Hz, 2H), 3.98 (s, 3H), 3.83-3.51 (m, 3H), 3.29 (m, 1H), 3.04 (m, 1H), 2.89 (m, 2H), 2.82-2.70 (m, 3H), 2.22 (d, J = 7.2 Hz, 3H), 2.18 (s, 3H), 2.13 (d, J = 10.2 Hz, 4H), 1.99-1.78 (m, 5H), 1.65 (d, J = 9.6 Hz, 3H), 1.50 (m, 1H), 1.46-1.32 (m, 4H), 1.13-1.00 (m, 2H). Example 66 [00657] N-(2-(1-(3-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro [5.5] undec-9-yl) methyl) piperidin-4-yl)-6- (difluoromethoxy)- 2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS-(ESI, m/z): [M + H] .sup.+ = 871.3. .sup.1H NMR (400 MHz, DMSO) δ 10.50 (s, 1H), 10.44 (s, 1H), 8.72 (s, 1H), 8.53 (s, 1H), 8.47 (d, J = 7.6 Hz, 1H), 8.41 (t, J = 7.7 Hz, 1H), 8.23 (d, J = 7.6 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.59-7.17 (m, 4H), 4.45 (m, 1H), 3.82-3.50 (m, 4H), 3.29 (m, 2H), 2.95 (m, 2H), 2.79- 2.71 (m, 2H), 2.18 (m, 2H), 2.08 (m, 6H), 1.71 (d, J = 8.5 Hz, 2H), 1.60 (m, 2H), 1.51 (m, 3H), 1.32 (d, J = 30.0 Hz, 2H), 1.10 (m, 4H). Example 67 [00658] N-(2-((1r,4r)-4- (((4-(1-(4-chloro- 3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl) piperidin-4-yl) butyl) (methyl) amino) methyl) cyclohexyl)-6- methoxy-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ESI, m/z): [M + H] .sup.+ = 851.3. .sup.1H NMR (400 MHz, DMSO) δ 10.50 (d, J = 2.0 Hz, 2H), 8.68 (s, 1H), 8.50-8.36 (m, 2H), 8.32 (s, 1H), 8.21 (d, J = 7.7 Hz, 1H), 7.61 (d, J = 8.2 Hz, 1H), 7.54 (d, J = 1.9 Hz, 1H), 7.37 (dd, J = 8.2, 2.0 Hz, 1H), 7.15 (s, 1H), 4.36 (t, J = 11.4 Hz, 2H), 3.98 (s, 3H), 3.80-3.70 (m, 1H), 3.65-3.50 (m, 2H), 3.12-2.92 (m, 1H), 2.80-2.64 (m, 3H), 2.35-2.20 (m, 2H), 2.20-2.08 (m, 6H), 1.98-1.46 (m, 9H), 1.45-1.35 (m, 2H), 1.35-1.20 (m, 4H), 1.15-1.01 (m, 4H). Example 68 [00659] N-(2-(1-((3-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6- (2-methoxyethoxy)- 2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ESI, m/z): [M + H] .sup.+ = 879.3. Example 69 [00660] N-(2-(1-((3-(4- chloro-3-(2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl)-3- azaspiro[5.5] undec-9-yl) methyl) piperidin-4-yl)-6- (oxetan-3-yloxy)- 2H-indazol-5- yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ESI, m/z): [M + H) .sup.+ = 877.3. Example 70 [00661] N-(2-((1r,4r)-4- (((1-(4-chloro-3- (2, 4-dioxotetrahydro- pyrimidin-1 (2H)-yl) benzoyl) piperidin-4-yl) methoxy) methyl) cyclohexyl)-6- methoxy-2H-indazol- 5-yl)-6-(trifluoro- methyl) pyridinecarbox- amide LC-MS: (ESI, m/z): [M + H] .sup.+ = 796.2. .sup.1H NMR (400 MHz, DMSO) δ 10.50 (s, 2H), 8.68 (s, 1H), 8.50- 8.36 (m, 2H), 8.32 (s, 1H), 8.22 (d, J = 7.6 Hz, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 1.9 Hz, 1H), 7.39 (dd, J = 8.2, 1.9 Hz, 1H), 7.16 (s, 1H), 4.39 (dd, J = 31.0, 19.6 Hz, 2H), 3.98 (s, 3H), 3.76 (d, J = 6.4 Hz, 1H), J = 5.9 Hz, 4H), 3.15-2.85 (m, 1H), 2.75 (dd, J = 10.9, 6.6 Hz, 3H), 2.14 (d, J = 9.1 Hz, 2H), 1.88 (t, J = 11.5 Hz, 5H), 1.80- 1.68 (m, 1H), 1.69-1.55 (m, 2H), 1.20 (dd, J = 25.5, 11.5 Hz, 4H).

Example 71: methyl (3-(2-chloro-5-(9-(2-(4-(6-(2-hydroxyprop-2-yl)-5-(6-(trifluoromethyl) pyridinecarboxamido)-2H-indazol-2-yl) piperidin-1-yl) ethyl)-3-azaspiro[5.5]undecyl-3-carbonyl) phenyl)-2, 6-dioxotetrahydropyrimidin-1 (2H)-yl) pivalate

[1815] ##STR00662##

[1816] Cesium carbonate (82 mg, 0.249 mmol) and chloromethyl pivalate (63 mg, 0.416 mmol) were added to a mixture of N-(2-(1-(2-(3-(4-chloro-3-(2, 4-dioxotetrahydropyrimidin-1 (2H)-yl) benzoyl)-3-azaspiro [5.5] undec-9-yl) ethyl) piperidin-4-yl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (182 mg, 0.208 mmol) in acetonitrile (5 mL). The reaction mixture was stirred at room temperature for 4h, filtered, then the filtrate was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The concentrate was purified by preparative HPLC to obtain 50 mg of target product as a light yellow solid.

[1817] LC-MS: (ESI, m/z): [M+H].sup.+=991.1.

[1818] .sup.1H NMR (400 MHz, DMSO) δ 12.37 (s, 1H), 8.71 (s, 1H), 8.45 (d, J=7.9 Hz, 1H), 8.40-8.33 (m, 2H), 8.16 (d, J=7.8 Hz, 1H), 7.66 (d, J=8.2 Hz, 1H), 7.61-7.54 (m, 2H), 7.42 (d, J=8.3 Hz, 1H), 5.94 (s, 1H), 5.69 (s, 2H), 4.49-4.36 (m, 1H), 3.84-3.63 (m, 2H), 3.62-3.50 (m, 2H), 3.30-3.20 (m, 2H), 2.97 (dt, J=16.7, 7.9 Hz, 4H), 2.40-2.35 (m, 2H), 2.10-1.96 (m, 6H), 1.75-1.67 (m, 2H), 1.62 (s, 6H), 1.58-1.47 (m, 3H), 1.47-1.32 (m, 4H), 1.31-1.22 (m, 2H), 1.18-1.04 (m, 13H).

Control group 1: N-(2-(1-(2-(2-((2-(2, 6-dioxopiperidin-3-yl)-1, 3-dioxoisoindol-5-yl) oxy) ethoxy) ethyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: Preparation of 2-(2, 6-dioxopiperidin-3-yl)-5-(2-(2-hydroxyethoxy) ethoxy) isoindolin-1, 3-dione

[1819] ##STR00663##

[1820] N,N-diisopropylethylamine (706.2 mg, 0.0055 mol) and potassium iodide (30 mg, 0.0002 mol) were added dropwise to a mixture of 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindolin-1,3-dione (500 mg, 0.0018 mol) and 2-(2-chloroethoxy)ethyl-1-ol (227 mg, 0.0018 mol) in dimethylsulfoxide (20 mL). The reaction solution was stirred at 100° C. for 10h. Water (30 ml) and ethyl acetate (100 ml) were added, the water layer was separated, and extracted with ethyl acetate (100 ml×2). The organic phases were combined and washed with saline (100 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 612 mg of target product as a white solid.

[1821] LC-MS: (ES, m/z): [M+H].sup.+=363.0

Step 2: Preparation of 2-(2-((2-(2, 6-dioxopiperidin-3-yl)-1, 3-dioxoisoindolin-5-yl) oxy) ethoxy) acetaldehyde

[1822] ##STR00664##

[1823] Dess-martin (393 mg, 0.88 mmol) was added to a mixture of 2-(2, 6-dioxopiperidin-3-yl)-5-(2-(2-hydroxyethoxy) ethoxy) isoindolin-1, 3-dione (162 mg, 0.44 mmol) in tetrahydrofuran (10 mL) at 0° C., and the obtained reaction solution was stirred for 16 hours at room temperature. The reaction solution was quenched with water (30 ml) and dichloromethane (50 ml). The water layer was separated and extracted with dichloromethane (50 ml/2). The organic phases were combined, washed with saturated saline (50 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain 32 mg of target product as a white solid.

[1824] LC-MS: (ES, m/z): [M+H].sup.+=361.0

Step 3: preparation of N-(2-(1-(2-(2-((2-(2, 6-dioxopiperidin-3-yl)-1, 3-dioxoisoindol-5-yl) oxy) ethoxy) ethyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1825] ##STR00665##

[1826] Sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added to a mixture of N-(6-methoxy-2-(piperidin-4-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide (24 mg, 0.05 mmol) and 2-(2-((2-(2, 6-dioxopiperidin-3-yl)-1, 3-dioxoisoindol-5-yl) oxy) ethoxy) acetaldehyde (40 mg, 0.11 mmol) in 1, 2-dichloroethane (5 ml). The reaction solution was stirred at room temperature for 2h. The solvent was removed by concentration under reduced pressure, and the concentrate was purified by HPLC to obtain 3.25 mg of target product as a white solid.

[1827] LC-MS: (ES, m/z): [M+H].sup.+=764.3

[1828] .sup.1H-NMR-LT-002-007: .sup.1H NMR (400 MHz, CD.sub.3OD_SPE) δ 8.77 (s, 1H), 8.47 (d, J=8.0 Hz, 3H), 8.31 (t, J=7.8 Hz, 1H), 8.18 (s, 1H), 8.05 (d, J=7.4 Hz, 1H), 7.81 (d, J=8.3 Hz, 1H), 7.47 (d, J=1.9 Hz, 1H), 7.35 (d, J=8.3 Hz, 1H), 7.07 (s, 1H), 5.12-5.07 (m, 1H), 4.51 (s, 1H), 4.37 (s, 2H), 4.05 (s, 3H), 3.91 (s, 2H), 3.82 (t, J=5.0 Hz, 2H), 3.37 (s, 2H), 2.96 (s, 2H), 2.90-2.77 (m, 1H), 2.77-2.60 (m, 4H), 2.26 (s, 4H), 2.08 (d, J=5.2 Hz, 1H).

Control group 2: N-(2-(1-((1-(2-((2-(2, 6-dioxopiperidin-3-yl)-1, 3-dioxoisoindol-5-yl) oxy) ethyl) piperidin-4-yl) methyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

Step 1: Preparation of 5-(allyloxy)-2-(2, 6-dioxopiperidin-3-yl) isoindolin-1, 3-dione

[1829] ##STR00666##

[1830] Under nitrogen protection, 3-bromoprop-1-ene (470 mg, 3.89 mmol) was added to a mixture of 2-(2, 6-dioxopiperidin-3-yl)-5-hydroxyisoindolin-1, 3-dione (1 g, 3.65 mmol) and potassium carbonate (1.01 g, 7.29 mmol) in N,N-dimethylformamide (10 mL). The reaction mixture was stirred overnight at 50° C. At 0° C., the reaction was quenched with water (60 mL) and extracted with ethyl acetate (30 mL×3). The organic phase was collected and washed with saturated saline, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure.

[1831] The concentrate was purified by column chromatography to obtain 0.85 g of the target product.

[1832] LC-MS: (ES, m/z): [M+H].sup.+=315.1

Step 2:2-((2-(2,6-dioxopiperidin-3-yl)-1, 3-dioxoisoindolin-5-yl) oxy) acetaldehyde

[1833] ##STR00667##

[1834] Ozone was introduced into a solution of 5-(allyloxy)-2-(2, 6-dioxopiperidin-3-yl) isoindolin-1, 3-dione (0.7 g, 2.23 mmol) in methylene chloride (200 ml) at −78° C. until the reaction solution turned to be blue. Then nitrogen was introduced until the reaction solution became colorless. Dimethyl sulfide (7.61 g, 123 mmol) was added at −78° C. under nitrogen protection, and the reaction mixture was stirred at room temperature overnight. The reaction solution was concentrated, and the concentrate was purified by column chromatography to obtain the target product.

[1835] LC-MS: (ES, m/z): [M+H].sup.+=317.1

Step 3: Preparation of N-(2-(1-((1-(2-((2-(2, 6-dioxopiperidin-3-yl)-1, 3-dioxoisoindol-5-yl) oxy) ethyl) piperidin-4-yl) methyl) piperidin-4-yl)-6-methoxy-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1836] ##STR00668##

[1837] It was synthesized with reference to the method of step 3 of control group 1.

[1838] LC-MS: (ESI, m/z): [M+H].sup.+=817.1.

[1839] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.11 (s, 1H), 10.50 (s, 1H), 8.69 (s, 1H), 8.46 (d, J=7.6 Hz, 1H), 8.41 (t, J=7.8 Hz, 1H), 8.37 (s, 1H), 8.22 (d, J=8.6 Hz, 1H), 7.83 (d, J=8.3 Hz, 1H), 7.47 (d, J=2.2 Hz, 1H), 7.37 (dd, J=8.3, 2.3 Hz, 1H), 7.16 (s, 1H), 5.12 (dd, J=12.8, 5.3 Hz, 1H), 4.43-4.34 (m, 1H), 4.29 (t, J=5.5 Hz, 2H), 3.00-2.91 (m, 5H), 2.78-2.70 (m, 2H), 2.64-2.54 (m, 2H), 2.22-2.16 (m, 3H), 2.13-2.02 (m, 9H), 1.74-1.66 (m, 2H), 1.58-1.44 (m, 1H), 1.18-1.08 (m, 2H).

Control group 3: N-(2-((1r,4r)-4-((((1-(2-(2, 6-dioxopiperidin-3-yl)-1, 3-dioxoisoindolin-4-yl) piperidin-4-yl) methyl) (methyl) amino) methyl) cyclohexyl)-6-(2-hydroxyprop-2-yl)-2H-indazol-5-yl)-6-(trifluoromethyl) pyridinecarboxamide

[1840] ##STR00669##

[1841] It was prepared with reference to the method of compound 1-317 in WO2020113233.

[1842] LC-MS: (ESI, m/z): [M+H].sup.+=843.3

[1843] .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.36 (s, 1H), 11.08 (s, 1H), 8.71 (s, 1H), 8.45 (d, J=7.8 Hz, 1H), 8.41-8.32 (m, 2H), 8.16 (d, J=7.8 Hz, 1H), 7.74-7.62 (m, 1H), 7.57 (s, 1H), 7.42-7.17 (m, 2H), 5.94 (s, 1H), 5.09 (dd, J=12.8, 5.3 Hz, 1H), 4.47-4.36 (m, 1H), 3.70 (d, J=10.0 Hz, 2H), 2.90 (d, J=12.7 Hz, 3H), 2.65-2.52 (m, 2H), 2.25-2.10 (m, 9H), 2.05-1.81 (m, 7H), 1.74-1.55 (m, 8H), 1.32 (d, J=11.6 Hz, 2H), 1.11 (d, J=11.9 Hz, 2H)

II Biological Activity Test Example

Test Example 1: IRAK4 Kinase Activity Test

[1844] KinEASE-STK S1 serine/threonine kinase kit (Cisbio) was used to detect the inhibitory effect of the compound on IRAK4 kinase activity. The specific method was: the compound was dissolved in dimethyl sulfoxide, and then it was diluted with the buffer solution of the kit by equal gradient to make the final concentration range of the tested compound in the reaction system 10000 nM-0.038 nM, then 2.5 nM kinase, 1 μM biotinylated polypeptide substrate and 7 μM adenosine triphosphate (ATP) were added in sequence and incubated at 37° C. for 120 min. Subsequently, anti-phosphorylated serine/threonine antibody coupled with europium element compound and modified XL665 streptavidin were added to the reaction system to terminate the reaction. After incubating at room temperature for 1 h, the fluorescence intensity of each well at the emission wavelength of 620 nm and 665 nm and the excitation wavelength of 337 nm was determined in HTRF mode on the microplate reader EnVision(PerkinElmer), the ratio value was calculated using the formula Ratio=(665 nm/620 nm)×10.sup.4. By comparing with the fluorescence intensity ratio of the control group, the inhibition rate of the compound at each concentration was calculated, and then the IC.sub.50 value of the compound was obtained by fitting the nonlinear curve with the logarithmic concentration-inhibition rate by GraphPad Prism 7.

TABLE-US-00023 TABLE 1 Example IC.sub.50(nM) Example 1 2.5 Example 2 2.9 Example 3 2.7 Example 4 12.2 Example 5 9.1 Example 6 7.6 Example 7 5.9 Example 8 10.5 Example 9 18.0 Example 10 6.5 Example 11 6.0 Example 12 7.9 Example 13 7.9 Example 14 6.9 Example 15 6.0 Example 16 9.2 Example 17 7.9 Example 18 9.6 Example 19 16.9 Example 20 9.5 Example 21 18.0 Example 22 11.7 Example 23 8.5 Example 24 34.6 Example 25 13.7 Example 26 6.5 Example 27 13.8 Example 28 16.2 Example 29 8.6 Example 30 16.0 Example 31 14.0 Example 32 28.5 Example 33 17.2 Example 34 16.5 Example 35 10.3 Example 36 11.8 Example 37 30.5 Example 38 34.5 Example 39 16.7 Example 40 35.1 Example 41 28.1 Example 42 39.2 Example 43 24.2 Example 44 52.4 Example 45 23.5 Example 46 43.1 Example 47 34.2 Example 48 61.8 Example 49 3.4 Example 50 7.2 Example 51 11.3 Example 52 9.2 Example 53 6.3 Example 54 7.5 Example 55 10.6 Example 56 3.6 Example 57 3.8 Example 58 4.3 Example 59 11.04 Example 60 6.43 Example 61 128.2 Example 62 6.9 Example 63 4.4 Example 64 48.0 Example 65 1.6 Example 66 21.4 Example 67 5.13 Example 71 114.0 Control group 1 44.2 Control group 2 15.2 Control group 3 16.2

[1845] Experimental results: The compound of the invention can effectively bind with the target protein and inhibit the IRAK4 kinase activity.

Test Example 2: Degradation of IRAK4 in THP-1 Cells by Compounds

[1846] Each well of the 24-well cell culture plate was inoculated with 0.95 mL THP-1 cells (stem cell bank of Chinese academy of sciences) with a cell density of 5×10.sup.5 cells/well. The cell plate was placed in a 5% carbon dioxide incubator and cultured overnight at 37° C., then 50 μL compound in dimethyl sulfoxide was added. The final concentration of the compound was in the range of 1-3000 nM. After continuous cultivation for 24 hours, the cells were collected into a 1.5 mL centrifuge tube, centrifuged at 1000 rpm and 4° C. for 5 minutes. Cell precipitation was cleaned twice with 1-DPBS, resuspended cells were lysed with 200 μL lysate (cell lysate was Western and IP cell lysate (Beyotime), supplemented with 1 mM mixture of phenylmethylsulfonyl fluoride and protease inhibitor (Beyotime)), left on ice for 30 minutes, then centrifuged at 14000 g at 4° C. for 10 minutes, and the supernatant was taken to detect IRAK4 protein level by Western blot.

[1847] Western blot method: The total protein concentration in the supernatant of cell lysis was determined by BCA protein quantitative kit (Tiangen). According to the concentration of total protein detected by BCA, the supernatant was adjusted to 0.2 μg/μL with PBS and 5× SDS-PAGE protein loading buffer (Beyotime), bathed in water at 100° C. for 10 minutes, then placed in ice bath for 5 minutes, centrifuged at 14000 g and 4° C. for 5 minutes, and then collected as WB loading sample. Prefabricated glue (KeyGEN) was used for protein electrophoresis with a loading amount of 10 μL (total protein 2 μg). After Tris-MOPS-SDS electrophoresis solution (Adamas), 120V constant pressure electrophoresis was performed. After electrophoresis, the protein on the adhesive strip was transferred to PVDF membrane with a constant current of 250V for 50 minutes. After the membrane was transferred, the membrane was placed in 1×QuickBlock blocking buffer (Beyotime) and incubated for 30 minutes at room temperature. After blocking, PVDF membranes were incubated with IRAK4 primary antibody (Abcam) overnight at 4°, the membranes were washed with TBST buffer (2.4 g Tris, 8.8 g NaCl, 1.5 mL Tween 20, pH adjusted to 7.4, constant volume to 1 L) for 30 minutes, incubated with secondary antibody (Abcam) for 2 hours at room temperature, finally incubated with Clarity Western ECL Substrate(BIO-RAD) for 5 minutes for luminescence development, chemiluminescence imaging system (Clinx, chemiScope 6200 Touch) for development and protein mapping photography. The protein map was analyzed by Clinx chemiluminescence analysis software for grayscale values. Using the formula: grayscale correction value=(target protein grayscale value/corresponding internal reference grayscale value)×10.sup.3, the grayscale correction value of each sample was calculated. The degradation rate was calculated by comparing with the grayscale correction value of the control group. Furthermore, DC.sub.50 and D.sub.max values of the compound were obtained by the nonlinear curve fitting with the logarithmic concentration-inhibition rate by GraphPad Prism 7.

TABLE-US-00024 TABLE 2 Example DC.sub.50(nM) D.sub.max(%) Example 1 3.0 94.6 Example 2 7.4 99.1 Example 3 23.1 90.5 Example 6 24.0 91.7 Example 7 9.4 95.5 Example 9 16.4 94.3 Example 10 2.3 100.0 Example 11 27.7 99.6 Example 12 1.4 96.9 Example 16 7.1 99.4 Example 17 5.3 96.9 Example 18 7.1 99.4 Example 19 13.4 92.8 Example 20 12.2 93.7 Example 21 4.0 90.1 Example 22 16.4 94.3 Example 23 32.4 97.6 Example 25 0.82 98.0 Example 26 1.0 86.9 Example 27 3.5 96.3 Example 28 0.8 99.4 Example 30 2.3 98.4 Example 31 2.6 97.1 Example 32 5.1 96.9 Example 33 2.1 96.6 Example 34 2.5 98.1 Example 35 8.4 95.2 Example 36 4.0 92.5 Example 39 10.5 89.4 Example 40 4.8 98.0 Example 41 2.9 97.7 Example 42 7.8 100.0 Example 43 4.9 98.7 Example 44 2.3 98.5 Example 45 1.9 98.2 Example 46 4.0 97.8 Example 47 5.4 98.3 Example 48 6.0 98.0 Example 49 1.8 99.2 Example 50 5.6 97.0 Example 51 27.5 95.4 Example 53 5.9 96.3 Example 54 1.4 98.9 Example 55 2.70 98.1 Example 56 3.0 97.0 Example 58 7.6 92.1 Example 60 5.1 93.2 Example 62 5.6 92.3 Example 64 13.9 95.6 Example 65 13.4 98.3 Example 66 16.1 95.3 Example 67 11.5 99.2 Example 71 3.70 97.0 Control group 1 38.1 73.9 Control group 2 40.6 85.1 Control group 3 19.8 75.3

[1848] Experimental results: The compound of the present invention can effectively degrade IRAK4 kinase protein in THP-1 cells and has excellent degradation activity.

Test Example 3: Degradation of IKZF1 and IKZF3 in L363 Cells by Compound

[1849] The 24-well cell culture plate was inoculated with 0.95 mL L363 cells (Nanjing Co-bioer) per well with a cell density of 6×10.sup.5 cells/well. The cell plate was placed in a 5% carbon dioxide incubator and cultured at 37° C. for 8 hours, then 50 μL of compound solution was added. The final concentration of the compound was 1000 nM. After continuing to culture for 16 hours, the cells were collected into a 1.5 mL centrifuge tube and centrifuged at 3000 rpm and 4° C. for 5 minutes. Cell precipitation was cleaned twice with 1×DPBS, resuspended cells were lysed with 100 μL lysate (cell lysate was Western and IP cell lysate (Beyotime), supplemented with 1 mM mixture of phenylmethylsulfonyl fluoride and protease inhibitor (Beyotime)), left on ice for 30 minutes, then centrifuged at 14000 g at 4° C. for 10 minutes, and the supernatant was taken to detect IKZF1 and IKZF3 protein level by Western blot.

[1850] The total protein concentration in the supernatant of cell lysis was determined by BCA protein quantitative kit (Tiangen). According to the concentration of total protein detected by BCA, the supernatant was adjusted to 0.2 μg/μL with PBS and 5× SDS-PAGE protein loading buffer (Beyotime), bathed in water at 100° C. for 10 minutes, then placed in ice bath for 5 minutes, centrifuged at 14000 g and 4° C. for 5 minutes, and then collected as WB loading sample. Prefabricated glue (KeyGEN) was used for protein electrophoresis with a loading amount of 10 μL (total protein 2 μg). After Tris-MOPS-SDS electrophoresis solution (Adamas), 120V constant pressure electrophoresis was performed. After electrophoresis, the protein on the adhesive strip was transferred to PVDF membrane with a constant current of 250V for 50 minutes. After the membrane was transferred, the membrane was incubated in 5% bovine serum albumin solution for 2 hours at room temperature. After blocking, PVDF membranes were incubated with IKZF1 primary antibody (CST) and IKZF3 primary antibody (Abcam) overnight at 4°, the membranes were washed with TBST buffer (2.4 g Tris, 8.8 g NaCl, 1.5 mL Tween 20, pH adjusted to 7.4, constant volume to IL) for 30 minutes, incubated with secondary antibody (Abcam) for 2 hours at room temperature, finally incubated with Clarity Western ECL Substrate(BIO-RAD) for 5 minutes for luminescence development, chemiluminescence imaging system (Clinx, chemiScope 6200 Touch) for development and protein mapping photography. The protein map was analyzed by Clinx chemiluminescence analysis software for grayscale values. Using the formula: grayscale correction value=(target protein grayscale value/corresponding internal reference grayscale value)×10.sup.3, the grayscale correction value of each sample was calculated. The degradation rate was calculated by comparing with the grayscale correction value of the control group.

TABLE-US-00025 TABLE 3 IKZF1 IKZF3 Concentration degradation rate degradation rate Compound (nM) (%) (%) Pomalidomide 1000 99.7 ± 0.2 98.6 ± 0.6 Control group 2 1000 47.5 ± 9.7 71.2 ± 5.7 Control group 3 1000 97.2 ± 2.6 92.8 ± 5.0 Example 34 1000 10.2 ± 1.8 −2.2 ± 2.8 Example 31 1000  8.4 ± 13.9 17.8 ± 3.6 Example 40 1000 53.8 ± 2.0  9.8 ± 2.0 Note: In Table 3, “—” refers to no degradation of IKZF1 and IKZF3.

[1851] The experimental results show that the compound of the present invention has good degradation selectivity, no or a small amount of degradation to IKZF1 and IKZF3, and has small toxic side effects, which is beneficial to the patent medicine.

Test Example 4: Determination of Cytokine Concentration of LPS-Induced Human PBMC Secretion

[1852] Frozen human PBMC (Milestone, PB010C) was resuspended to RPMI 1640 medium (Gibco, A1049101) after resuscitation, supplemented with 10% fetal bovine serum (Gibco, 10099141), 100 U/mL penicillin and 100 μg/mL streptomycin (Gibco, 15140122). On the same day, 150 μL of PBMC was spread in each well of a 96-well transparent cell plate (Labserv, 310109008), so that the cell density was 2×105 cells/well. Then 50 μL diluted compound solution was added to the corresponding cell-covered wells to make the final concentration of the compound in the range of 0.026-10000 nM. The final concentration of DMSO was 0.25%. After the drug-treated cell plate was cultured at 37° C. for 1 hour or 20 hours in a 5% CO2 incubator, 10 μL LPS (0111:B4) (Sigma, L4391) was added with a final concentration of 100 ng/mL. The cell plate was again placed in a 5% CO2 incubator at 37° C. for 5 hours, centrifuged at 2000 rpm for 10 minutes, and 100 μL cell supernatant was taken from each well and transferred to a new 96-well transparent cell plate for cytokine concentration test, which was frozen at −80° C. to be tested.

[1853] Interleukin-6 (IL-6) AlphaLISA Detection

[1854] The concentration of IL-6 in the supernatant of LPS-induced human PBMC cells was determined using interleukin-6 (IL-6) AlphaLISA detection kit (PerkinElmer, AL223 C).

[1855] According to the product instructions, IL-6 standard solutions with different concentrations were prepared in the concentration range of 0-100000 μg/mL. 2 μL of IL-6 standard solution at various concentrations and the cell supernatant sample to be tested were taken, respectively added to white 384 well plates (PerkinElmer, 6007299), then a mixed solution of 8 μL of 5× anti-IL-6 receptor beads (final concentration: 10 μg/mL) and biotin-labeled anti-IL-6 antibody (final concentration 1 nM) was added to each well, and incubated at 23° C. for 60 minutes. Finally, 10 μL of 2× streptavidin-labeled donor beads (final concentration: 40 μg/mL) were added to each well and incubated at 23° C. for 30 minutes in the absent of light. After the incubation, the AlphaLISA signal value was determined in the AlphaScreen standard setting mode on the microplate reader EnVision(PerkinElmer, 2105). The standard curve was drawn by the AlphaLISA signal value of each concentration IL-6 standard solution, and then based on the corresponding concentration on the standard curve for the AlphaLISA signal value of the sample to be tested, the IL-6 concentration of each cell lysis supernatant was determined. Compared with the IL-6 concentration of the control group, the inhibition rate of the compound at each concentration was calculated, and then IC.sub.50 value of the compound was calculated by fitting the nonlinear curve with the logarithmic concentration-inhibition rate by GraphPad Prism 7.

TABLE-US-00026 TABLE 4 Example IC.sub.50(nM) Control group 2 2046 Control group 3 47.5 Example 1 4.9 Example 28 4.7 Example 31 1.8 Example 32 9.7 Example 41 5.1 Example 55 0.8 Example 53 2.8 Example 49 3.7 Example 54 2.6 Example 56 3.2

[1856] Experimental results: The compounds of the present invention have a good inhibitory effect on IL-6 production by immune cells.