WO2022002097A1 - Pyrazolo[1,5-a]pyridine compound, preparation method therefor and use thereof - Google Patents

Abstract

The present invention relates to a pyrazolo[1,5-a]pyridine compound as represented by the general formula (I), a preparation method therefor, a pharmaceutical composition, and the use thereof. In the formula, R1, R2, R3, R4, R5, X, Y, and Z are as defined in the description. The pyrazolo[1,5-a]pyridine compound of the present invention can effectively inhibit wild-type RET, mutant RET, specifically G810R mutant RET, and RET fusion, such as KIF5B-RET and CCDC6-RET, same has good safety, and can be used for treating RET-mediated diseases, such as cancers and irritable bowel syndrome.

Description

Pyrazo[1,5-a]pyridine compounds and preparation method and application thereof technical field

The invention belongs to the field of medicine, and in particular relates to pyrazo[1,5-a]pyridine compounds that can be used as RET inhibitors, a preparation method of such compounds, pharmaceutical compositions containing such compounds, such compounds and medicines thereof Use of a composition in the treatment of a disease mediated by RET.

Background technique

The RET (rearranged during transfection) gene is a proto-oncogene and is located on chromosome 10. The RET protein encoded by the RET gene is a receptor tyrosine kinase (RTK) that exists on the cell membrane and belongs to the cadherin superfamily. The ligands for RET are the glial-derived neurotrophic factor family, including glial cell line-derived neurotrophic factor (GDNF), neurturin (NRTN), artemin (ARTN), and persephin (PSPN). When the ligand binds to the extracellular domain of RET, it will lead to dimerization of RET and autophosphorylation of the intracellular kinase domain, thereby activating RET and then initiating multiple downstream signaling pathways such as RAS/MAPK/ERK, PI3K/AKT, JAK /STAT plays the role of cell proliferation, migration and differentiation.

Abnormalities of the RET gene (including fusions and point mutations) lead to excessive activation of the RET signaling pathway and uncontrolled cell growth, which are closely related to the occurrence and progression of various tumors. The incidence of RET gene fusions is approximately 1% to 2% in non-small cell lung cancer (NSCLC) patients and 10% to 20% in papillary thyroid cancer (PTC) patients; the most common fusion partners include KIF5B, TRIM33, CCDC6 and NCOA4. The incidence of RET gene point mutation in medullary thyroid cancer (MTC) patients is about 60%; common mutation sites include C634R, M918T, V804L, V804M, G810R, etc. In addition, low frequencies of RET fusions have also been observed in colorectal, breast, pancreatic, and other cancers; RET fusions have also been observed in NSCLC patients with EGFR mutations.

The current targeted drugs for the RET gene include (1) multi-kinase inhibitors targeting multiple targets, such as Cabozantinib, Vandetanib, Lenvatinib ), etc., although it can inhibit the activity of RET, but due to the low selectivity, off-target effects usually occur, especially serious side effects related to VEGFR inhibition, which limit the clinical application; (2) Drugs that selectively target the RET gene , such as BLU-667, LOXO-292, which have good inhibitory activity on wild-type and mutant (such as M918T, V804L/M, etc.) RET, as well as common KIF5B-RET fusion and CCDC6-RET fusion, etc. and relatively low toxicity.

BLU-667 was disclosed in WO2017079140A1 and was developed by Blueprint Medicines. Its structure is as follows, and a rolling application for new drug marketing has been submitted to the FDA. LOXO-292 was disclosed in WO2018071447A1 and developed by Loxo Oncology. Its structure is as follows. It was approved by the FDA in May 2020; It has been reported that patients with RET fusion NSCLC developed the RET G810R resistance mutation, resulting in LOXO-292 resistance.

In addition, abnormal RET expression and/or activity has also been demonstrated in gastrointestinal disorders such as irritable bowel syndrome (IBS).

The development of new potent, low toxicity and selective RET small-molecule inhibitors for the treatment of RET-mediated diseases is needed in humans.

SUMMARY OF THE INVENTION

The present invention provides a pyrazo[1,5-a]pyridine compound with novel structure or a pharmaceutically acceptable salt thereof, which is a kind of selective RET inhibitor, which can effectively inhibit wild-type RET, mutant RET and special It is a G810R mutant RET, and RET fusions such as KIF5B-RET and CCDC6-RET, which are safe and can be used for the treatment of RET-mediated diseases such as cancer and irritable bowel syndrome.

The present invention provides a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof,

in:

X is selected from CR ₆ or N;

Y is selected from CR ₆ or N;

Z is selected from CR ₆ or N;

where 0, 1 or 2 of X, Y and Z are N;

R _{1 is} selected from hydrogen, 6-10 membered aryl or 5-12 membered heteroaryl, wherein each of said 6-10 membered aryl or 5-12 membered heteroaryl is optionally replaced by 1, 2, 3 or 4 each independently selected from the group consisting of hydroxy, halogen, C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, hydroxy C _1-6 alkoxy substituted with substituents of -NR ₇ R ₈ or -O-(CH ₂ ) _n -NR ₇ R ₈ ;

R _{2 is} selected from hydrogen, C _1-4 alkyl or halogenated C _1-4 alkyl;

R _{3 is} selected from hydrogen, C _1-6 alkyl, 3-6 membered cycloalkyl, 3-8 membered heterocyclyl or 5-12 membered heteroaryl, wherein said C _1-6 alkyl, 3-6 membered cycloalkyl, 3-8 membered heterocyclyl or 5-12 membered heteroaryl group each optionally substituted with two, three or four substituents each independently selected from halogen, C _1-4 alkyl, C _{1- 4} alkoxy, halogenated C _1-4 alkyl, halogenated C _1-4 alkoxy, cyano, hydroxy, hydroxy C _1-4 alkyl, -C(=O)-NR ₇ R ₈ , - SO ₂ -C _1-4 alkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclyl substituent, wherein the 5-6 membered heteroaryl or 3-6 membered heterocyclyl are each optionally by 1, 2 or 3 each independently selected from halogen, hydroxy, C _1-4 alkyl, C _1-4 alkoxy-(CH ₂ ) _m - or -C(=O)-C _{1 -4} alkyl substituents are substituted;

R _{4 is} selected from hydrogen, C _1-4 alkyl or halogenated C _1-4 alkyl;

R _{5 is} selected from hydrogen, halogen, cyano, C _1-4 alkyl or halogenated C _1-4 alkyl;

each R _{6 is} independently selected from H, C _1-4 alkyl, C _1-4 alkoxy, haloC _1-4 alkyl or halogen;

each R _{7 is} independently selected from H, C _1-4 alkyl or halogenated C _1-4 alkyl;

each R _{8 is} independently selected from H, C _1-4 alkyl or halogenated C _1-4 alkyl;

n is selected from 1, 2 or 3;

m is selected from 0, 1, 2 or 3.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above-mentioned X is selected from CR ₆ or N, Y is selected from CR ₆ or N, Z is selected from CR ₆ or N, wherein 0 or 1 of X, Y, Z is N; wherein each R _{6 is} independently selected from H, C _1-4 alkyl, haloC _1-4 alkyl or halogen.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above-mentioned X is CR ₆ , Y is CR ₆ , and Z is CR ₆ ; wherein each R _{6 is} independently selected from H, C _1-4 alkyl, halogenated C _1-4 alkyl or halogen.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above-mentioned X is CR ₆ , Y is CR ₆ , and Z is CR ₆ ; wherein each R _{6 is} independently selected from H, methyl, ethyl, monofluoromethyl, difluoromethyl, _{trifluoromethyl, CF 3 CH 2 -, CHF} 2 CH 2 -, CH 2 FCH 2 -, F, Cl or Br.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above-mentioned X is CR ₆ , Y is CR ₆ , Z is CR ₆ , wherein each R ₆ is H.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{1 is} selected from phenyl or 5-6 membered heteroaryl, wherein the phenyl or 5-6 Each of the membered heteroaryl groups is optionally replaced by 1, 2 or 3 each independently selected from the group consisting of hydroxy, halogen, _C1-6 alkyl, _C1-6 alkoxy, halogenated _C1-6 alkyl, halogenated C _1-6 alkoxy or cyano substituent.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{1 is} selected from phenyl or 5-6 membered heteroaryl, wherein the phenyl or 5-6 membered heteroaryl group each optionally substituted with 1, 2 or 3 substituents each independently selected from hydroxy, halo, C _1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl or halo Substituents of C _{1-4 alkoxy.}

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{1 is} selected from phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyrrolyl, imidazolyl or thiazole wherein the phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyrrolyl, imidazolyl, or thiazolyl groups are each optionally surrounded by 1, 2 or 3 each independently selected from hydroxy, halogen, C _1-4 Substituents of alkyl, C _1-4 alkoxy, halogenated C _1-4 alkyl or halogenated C _{1-4 alkoxy.}

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{1 is} selected from phenyl,

wherein the phenyl,

Each optionally substituted with 1, 2 or 3 substituents each independently selected from hydroxy, F, Cl, Br, C 1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl or halo Substituents of C _{1-4 alkoxy.}

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{1 is} selected from phenyl,

wherein the phenyl,

Each is optionally selected by 1, 2 or 3 each independently selected from hydroxy, F, Cl, Br, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, n-propoxy , isopropoxy, monofluoromethyl, difluoromethyl, trifluoromethyl, CF ₃ CH ₂ -, CHF ₂ CH ₂ -, CH ₂ FCH ₂ -, monofluoromethoxy, difluoromethoxy , trifluoromethoxy, CF ₃ CH ₂ -O-, CHF ₂ CH ₂ -O- or CH ₂ FCH ₂ -O- substituents.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{1 is} selected from phenyl or pyridyl, wherein each of said phenyl or pyridyl is optionally replaced by 1 , 2 or 3 substituents each independently selected from hydroxy, halogen, C _1-4 alkyl, C _1-4 alkoxy, halogenated C _1-4 alkyl or halogenated C _1-4 alkoxy replaced.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{1 is} selected from phenyl,

wherein the phenyl,

Each is optionally selected by 1, 2 or 3 each independently selected from hydroxy, F, Cl, Br, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, n-propoxy , isopropoxy, monofluoromethyl, difluoromethyl, trifluoromethyl, CF ₃ CH ₂ -, CHF ₂ CH ₂ -, CH ₂ FCH ₂ -, monofluoromethoxy, difluoromethoxy , trifluoromethoxy, CF ₃ CH ₂ -O-, CHF ₂ CH ₂ -O- or CH ₂ FCH ₂ -O- substituents.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{1 is} selected from

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{1 is} selected from

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{2 is} selected from hydrogen or C _1-4 alkyl.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R ₂ is hydrogen.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{4 is} selected from hydrogen or C _1-4 alkyl.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R ₄ is hydrogen.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{5 is} selected from hydrogen, halogen or cyano.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R ₅ is cyano.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{3 is} selected from hydrogen, C _1-6 alkyl or 3-6 membered cycloalkyl, wherein the Each of C _1-6 alkyl or 3-6 membered cycloalkyl is optionally substituted by 1, 2 or 3 each independently selected from halogen, C _1-4 alkyl, C _1-4 alkoxy, halogenated C _{Substituents of 1-4} alkyl, halogenated C _1-4 alkoxy, cyano, hydroxy, -SO ₂ -C _1-4 alkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclyl substituted, wherein each of the 5-6 membered heteroaryl or 3-6 membered heterocyclyl is optionally substituted with 1, 2 or 3 substituents each independently selected from halogen, hydroxy or C _1-4 alkyl replaced.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{3 is} selected from hydrogen, C _1-4 alkyl, cyclopropyl, cyclobutyl or cyclopentyl , wherein the C _1-4 alkyl, cyclopropyl, cyclobutyl or cyclopentyl groups are each optionally surrounded by 1, 2 or 3 each independently selected from F, Cl, Br, C _1-4 alkyl , C _1-4 alkoxy, halogenated C _1-4 alkyl, halogenated C _1-4 alkoxy, cyano, hydroxyl, -SO ₂ -C _1-4 alkyl,

substituted by the substituents in which the

Each is optionally substituted with 1, 2 or 3 substituents each independently selected from F, Cl, Br, hydroxy or _C1-4 alkyl.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{3 is} selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclopropyl, cyclobutyl or cyclopentyl, wherein said methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec The butyl, tert-butyl, cyclopropyl, cyclobutyl or cyclopentyl groups are each optionally selected by 1, 2 or 3 each independently selected from F, Cl, Br, methyl, ethyl, methoxy, ethoxy group, a fluoromethyl, difluoromethyl, _{trifluoromethyl, CF 3 CH 2 -, CHF} 2 CH 2 -, CH 2 FCH 2 -, a fluoromethoxy, difluoromethoxy, tris Fluoromethoxy, CF ₃ CH ₂ -O-, CHF ₂ CH ₂ -O-, CH ₂ FCH ₂ -O-, cyano, hydroxyl, -SO ₂ -CH ₃ , -SO ₂ -CH ₂ CH ₃ ,

substituted by the substituents in which the

Each is optionally substituted with 1, 2 or 3 substituents each independently selected from F, Cl, Br, hydroxy, methyl or ethyl.

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{3 is} selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl base, sec-butyl, tert-butyl, monofluoromethyl, difluoromethyl, trifluoromethyl,

In another embodiment of the compound represented by formula (I) of the present invention or a pharmaceutically acceptable salt thereof, the above R _{3 is} selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl base, sec-butyl, tert-butyl, monofluoromethyl, difluoromethyl, trifluoromethyl,

The present invention also provides the following compounds or their pharmaceutically acceptable salts:

The compound represented by the formula (I) of the present invention can be prepared by various synthetic methods well known to those skilled in the art without particular limitation. The method for synthesizing the compound represented by formula (I) of the present invention is not limited to Scheme 1 and Scheme 2, and those skilled in the art can perform conventional substitutions and improvements.

Option One:

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , X, Y, Z are as defined above; L is a leaving group, such as halogen, preferably I; P is an amino protecting group, each P can be The same or different, each independently preferably tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), benzyl (Bn).

Using compound a-1 and compound a-2 as starting materials, Suzuki coupling reaction occurs to obtain compound a-3; compound a-3 undergoes substitution reaction with compound a-4 to obtain intermediate 1; intermediate 1 is deprotected to obtain compound a-5; compound a-5 undergoes reductive amination reaction with compound a-6 to obtain intermediate 2; intermediate 2 is combined with

A coupling reaction occurs to obtain the compound represented by formula (I).

Alternatively, when preparing the _{compound of formula (I) in which R 4} is H, intermediate 2 can also _{undergo coupling reaction with P-NH 2} to obtain compound a-7; compound a-7 undergoes substitution reaction _{with R 3 -L to obtain compound a-7} Compound a-8; Compound a-8 is deprotected to obtain the compound represented by formula (I).

In the above scheme 1, the Suzuki coupling reaction is carried out in the presence of a base and a catalyst, wherein the catalyst includes but is not limited to Pd(dppf)Cl ₂ , Pd(PPh ₃ ) ₄ , PdCl _{2 ,} etc., the base includes but not limited to Limited to cesium carbonate, potassium carbonate, sodium carbonate, lithium carbonate, potassium acetate, sodium acetate, potassium tert-butoxide, sodium tert-butoxide, etc.; the substitution reaction of preparation intermediate 1 and compound a-8 is carried out in the presence of a base the bases include, but are not limited to cesium carbonate, potassium carbonate, sodium carbonate, triethylamine and the like; and intermediate compound 1 a-8 deprotection reaction is carried out at or catalyst / H ₂ in the presence of acidic conditions, the The acidic conditions include but are not limited to HCl/MeOH, HCl/EA, trifluoroacetic acid, etc., and the catalysts include but are not limited to Pd/C, PdCl ₂ /C, Pd(OH) ₂ /C, etc.; The reductive amination reaction is carried out in the presence of a reducing agent, and the reducing agent includes but is not limited to NaBH(OAc) ₃ , NaBH ₄ , NaBH ₃ CN, etc.;

or P-NH ₂ The coupling reaction that occurs is carried out in the presence of a catalyst and a base, and the catalyst includes but is not limited to Pd ₂ (dba) ₃ /Xantphos, Pd ₂ (dba) ₃ /Brettphos, Pd(OAc) ₂ / Xantphos, etc., the bases include but are not limited to cesium carbonate, potassium carbonate, sodium carbonate, lithium carbonate, potassium acetate, sodium acetate, potassium tert-butoxide, sodium tert-butoxide, and the like.

Option II:

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , X, Y, Z are as defined above; L is a leaving group, such as halogen, preferably I; P is an amino protecting group, each P can be The same or different, each independently preferably tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), benzyl (Bn).

Intermediate 1 with

Coupling reaction occurs to obtain compound b-1; compound b-1 is deprotected to obtain intermediate 3; intermediate 3 undergoes reductive amination reaction with compound a-6 to obtain the compound represented by formula (I).

Alternatively, when R ₄ is the preparation of a compound of the formula H (the I), may be reacting intermediate 1 P-NH ₂ to give compound b-2 and the coupling reaction; b-2 with the compound R ₃ -L substitution reaction to give Compound b-3; compound b-3 is deprotected to obtain intermediate 3; intermediate 3 is subjected to reductive amination reaction with compound a-6 to obtain the compound represented by formula (I).

In the above scheme two, intermediate 1 and

or P-NH ₂ The coupling reaction that occurs is carried out in the presence of a catalyst and a base, and the catalyst includes but is not limited to Pd ₂ (dba) ₃ /Xantphos, Pd ₂ (dba) ₃ /Brettphos, Pd(OAc) ₂ / Xantphos, etc., the bases include but are not limited to cesium carbonate, potassium carbonate, sodium carbonate, lithium carbonate, potassium acetate, sodium acetate, potassium tert-butoxide, sodium tert-butoxide, etc.; protection or in the catalyst / H ₂ in the presence of acidic conditions, the acidic conditions include, but are not limited to HCl / MeOH, HCl / EA, trifluoroacetic acid, etc., the catalysts include, but are not limited to Pd / C, PdCl ₂ /C, Pd(OH) ₂ /C, etc.; the reductive amination reaction between intermediate 3 and compound a-6 is carried out in the presence of a reducing agent, including but not limited to NaBH(OAc) ₃ , NaBH ₄ , NaBH ₃ CN, etc.; the substitution reaction for preparing compound b-3 is carried out in the presence of a base, and the base includes but is not limited to cesium carbonate, potassium carbonate, sodium carbonate, triethylamine and the like.

The present invention also provides intermediate compounds, including compound a-7, compound a-8, compound b-1, compound b-2, compound b-3, and intermediate 3 shown in the following structures,

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , X, Y, Z are as defined above; P is an amino protecting group, each P may be the same or different, and each independently preferably tert-butoxycarbonyl (Boc ), benzyloxycarbonyl (Cbz), benzyl (Bn).

The present invention further provides the intermediate compound of the following structure:

The present invention also provides a pharmaceutical composition comprising the above-mentioned compound or a pharmaceutically acceptable salt thereof and optionally a pharmaceutically acceptable carrier. The pharmaceutical composition of the present invention including optional pharmaceutically acceptable carrier means that the composition may or may not contain a pharmaceutically acceptable carrier.

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof in the preparation of a medicament for treating a disease mediated by RET.

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof in the preparation of a medicament for treating a disease mediated by RET. Wherein the RET includes wild-type RET, mutant RET, RET fusion, and the mutant RET includes but is not limited to G810R mutant RET, M918T mutant RET, V804L mutant RET, V804M mutant RET, preferably G810R mutant RET , the RET fusion includes but is not limited to KIF5B-RET fusion, CCDC6-RET fusion, preferably KIF5B-RET fusion; the diseases include cancer and irritable bowel syndrome.

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof in the preparation of a medicament for the treatment of diseases mediated by wild-type RET, mutant RET, or RET fusion; wherein the mutation Type RET includes but is not limited to G810R mutant RET, M918T mutant RET, V804L mutant RET, V804M mutant RET, preferably G810R mutant RET, and the RET fusion includes but is not limited to KIF5B-RET fusion, CCDC6-RET fusion, KIF5B-RET fusions are preferred; such diseases include cancer, irritable bowel syndrome.

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt or pharmaceutical composition thereof in the preparation of a medicament for the treatment of RET-mediated cancer and irritable bowel syndrome. Wherein the RET includes wild-type RET, mutant RET, RET fusion, and the mutant RET includes but is not limited to G810R mutant RET, M918T mutant RET, V804L mutant RET, V804M mutant RET, preferably G810R mutant RET , the RET fusion includes but is not limited to KIF5B-RET fusion, CCDC6-RET fusion, preferably KIF5B-RET fusion.

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt or pharmaceutical composition thereof in the preparation of a medicament for the treatment of cancer and irritable bowel syndrome mediated by wild-type RET, mutant RET, RET fusion application. Wherein the mutant RET includes but is not limited to G810R mutant RET, M918T mutant RET, V804L mutant RET, V804M mutant RET, preferably G810R mutant RET; the RET fusion includes but is not limited to KIF5B-RET fusion, CCDC6 -RET fusion, preferably KIF5B-RET fusion.

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt or pharmaceutical composition thereof in the preparation of a medicament for treating cancer.

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt or pharmaceutical composition thereof in the preparation of a medicament for treating irritable bowel syndrome.

The present invention also provides a method of treating a disease mediated by RET, the method comprising administering to a patient in need thereof an effective amount of the above-mentioned compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof. The RET includes wild-type RET, mutant RET, and RET fusion, and the mutant RET includes but is not limited to G810R mutant RET, M918T mutant RET, V804L mutant RET, and V804M mutant RET, and the RET fusion includes But not limited to KIF5B-RET fusion, CCDC6-RET fusion; the diseases include cancer, irritable bowel syndrome.

The present invention also provides a method of treating cancer, the method comprising administering to a patient in need thereof an effective amount of the above-mentioned compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof.

The present invention also provides a method of treating irritable bowel syndrome, the method comprising administering to a patient in need thereof an effective amount of the above-mentioned compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof.

Cancers described in the present invention include but are not limited to small cell lung cancer, non-small cell lung cancer, papillary thyroid cancer, medullary thyroid cancer, follicular thyroid cancer, anaplastic thyroid cancer, recurrent thyroid cancer, multiple endocrine neoplasia Type 2A or 2B (MEN2A or MEN2B, respectively), hepatocellular carcinoma, lung cancer, head and neck cancer, glioma, neuroblastoma, pheochromocytoma, colorectal cancer, testicular cancer, prostate cancer, fallopian tube cancer, ovarian cancer cancer, cervical cancer, breast cancer and pancreatic cancer.

The irritable bowel syndrome described in the present invention includes, but is not limited to, diarrhea-predominant, constipation-predominant or alternate defecation pattern, functional flatulence, functional constipation, functional diarrhea, non-specific functional bowel disease, and functional abdominal pain syndrome, chronic idiopathic constipation, functional esophageal disease, functional gastroduodenal disease, functional anorectal pain, and inflammatory bowel disease.

The present invention also includes pharmaceutically acceptable salts of the compounds represented by formula (I). The "pharmaceutically acceptable salts" refer to relatively nontoxic acid addition or base addition salts of the compounds of the present invention. The acid addition salt is the salt formed by the compound represented by the formula (I) of the present invention and a suitable inorganic acid or organic acid. These salts can be obtained by mixing the compound represented by the formula (I) with a suitable organic acid or inorganic acid. prepared by reacting in a solvent. Representative acid addition salts include hydrobromide, hydrochloride, sulfate, bisulfate, sulfite, acetate, oxalate, valerate, oleate, palmitate, stearic acid Salt, laurosilicate, borate, benzoate, lactate, nitrate, phosphate, hydrogen phosphate, carbonate, bicarbonate, toluate, citrate, maleic acid Salt, fumarate, succinate, malate, ascorbate, tannate, pamoate, alginate, naphthalene sulfonate, tartrate, benzoate, mesylate, p-toluene Sulfonate, gluconate, lactobionate and lauryl sulfonate, etc. The base addition salt is the salt formed by the compound represented by the formula (I) and a suitable inorganic base or organic base. These salts can be obtained by mixing the compound represented by the formula (I) with a suitable inorganic base or organic base in a suitable solvent. prepared by the reaction. Representative base addition salts include, for example, salts formed with alkali metal, alkaline earth metal, quaternary ammonium cations, such as sodium, lithium, potassium, calcium, magnesium, tetramethylquaternary ammonium, tetraethylquaternary ammonium Salts, etc.; amine salts, including salts formed with ammonia (NH3), primary amine, secondary amine or tertiary amine, such as methylamine salt, dimethylamine salt, trimethylamine salt, triethylamine salt, ethylamine salt, etc.

In addition to salt forms, the compounds provided herein also exist in prodrug forms. The prodrugs described in the present invention refer to compounds that can be converted into the biologically active compounds described in the present invention (eg, compounds of formula (I)) under physiological conditions or by solvolysis. Thus, the term "prodrug" refers to a pharmaceutically acceptable precursor of a biologically active compound.

The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, radiolabeled compounds can be used, such as deuterium (D), tritium ⁽³ H), iodine -125 ⁽¹²⁵ I) or ^C-14 (14 C). All transformations of the isotopic composition of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, and racemic mixtures thereof and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which belong to this within the scope of the invention. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.

Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting mixture of diastereomers is separated and the auxiliary group is cleaved to provide pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, followed by conventional methods known in the art The diastereoisomers were resolved and the pure enantiomers recovered. In addition, separation of enantiomers and diastereomers is usually accomplished by the use of chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (eg, from amines to amino groups) formate).

In the present invention, "pharmaceutically acceptable" means that, within the scope of sound medical judgment, it is suitable for contact with human and animal tissues without undue toxicity, irritation or other problems or complications, commensurate with a reasonable benefit/risk ratio those compounds, substances, compositions and dosage forms.

In the present invention, "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid filler or gel substances, which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. "Compatibility" as used herein means that the components of the composition can be blended with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Examples of pharmacologically acceptable carrier moieties are cellulose and its derivatives (such as sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, hypromellose and its derivatives, cellulose acetate and its derivatives) cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium/calcium stearate, hydrogenated vegetable oil, sodium fumarate stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers, wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, Antioxidants, preservatives, etc.

In the present invention, for a drug or a pharmacologically active agent, "effective amount" refers to a sufficient amount of the drug or pharmacologically active agent that is non-toxic but can achieve the desired effect. The determination of the effective amount varies from person to person, depending on the age, weight and condition of the patient, and also on the specific active substance. The appropriate effective amount in individual cases can be determined by those skilled in the art based on routine experiments.

In the present invention, "active ingredient", "active substance" or "active agent" refers to a chemical entity that is effective in treating the target disorder, disease or condition.

In the present invention, "patient", "individual" or "subject" includes humans, animals, vertebrates, mammals, rodents (eg, guinea pigs, hamsters, rats, mice), murine animals (eg, mice), Canines (eg, dogs), primates, great apes (eg, monkeys or apes), monkeys (eg, marmosets, baboons), apes (eg, gorillas, chimpanzees, orangutans, gibbons). In some embodiments, a "patient" is a human.

In the present invention, "treating" means administering a compound or formulation of the present invention to prevent, ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes:

(i) preventing the emergence of a disease or condition in mammals, particularly when such mammals are susceptible to the disease or condition but have not been diagnosed with the disease or condition;

(ii) inhibiting a disease or disease state, i.e., arresting its development;

(iii) alleviating the disease or disease state, ie, causing the disease or disease state to regress.

In the present invention, "optional", "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes both instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted with R" means that it may or may not be substituted with R, and the description includes both R substitution and non-R substitution.

In the present invention, "substituted" or "substituted" refers to one or more hydrogen atoms in the group, preferably 1-5 hydrogen atoms, more preferably 1, 2, 3 or 4 hydrogen atoms with each other are independently substituted with the corresponding number of substituents, as long as the substituted compound is stable.

In the present invention, "independently" or "independently" means wherein more than one substituent is selected from a large number of possible substituents, which may be the same or different. For example, "R _{x is} selected from -OR or -N(R) ₂ ; each R is independently selected from H or C ₁ -C ₄ alkyl", where each substituent R is independently, they may be the same or can be different.

In the present invention, "group" and "group" denote a monovalent group or a divalent or higher group corresponding to a valence as required, for example, "cycloalkyl" includes a monovalent group obtained by removing one hydrogen atom therefrom The group also includes divalent or more groups obtained by removing two or more hydrogen atoms from the same carbon atom or two or more different carbon atoms. When a "cycloalkyl" is used as a terminal group, it is naturally a monovalent group, and when a cycloalkyl group is used as a linking group in the structure, it is a divalent or higher group. In the present invention, a monovalent or more than divalent group usually refers to a monovalent group or a divalent group, but as required, the group can be a higher valence (such as trivalent, tetravalent, pentavalent, hexavalent, etc.).

In the present invention, "C _de " (d and e represent an integer of 1 or more, d<e) includes any specific case of d to e carbons, for example, C _1-6 includes C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , also include any one range from d to e, for example, C _1-6 includes C _1-3 , C _1-4 , C _1-5 , C _2-5 , C _2-4 , C _3-6, etc.; in the same way, "de element" (d and e represent an integer of 1 or more, d<e) indicates that the number of atoms on the ring is d to e, for example, a 3-6-membered ring includes a 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, and any range from d to e, for example, 3-6-membered ring includes 3-4-membered ring, 3-5-membered ring, 4-6-membered ring, 4- 5-membered ring, etc.

In the present invention, "halogen" refers to fluorine, chlorine, bromine, iodine and the like, preferably fluorine, chlorine and bromine, more preferably fluorine and chlorine.

In the present invention, "C _1-6 alkyl group" refers to a straight-chain or branched alkyl group derived from an alkane moiety containing 1-6 carbon atoms by removing one or more hydrogen atoms, specifically, C _1-6 Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl , neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3,3- Dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,3- Dimethylbutyl, 2-ethylbutyl, 1-methyl-2-methylpropyl, etc.; the "C _1-4 alkyl" refers to the moiety removed from an alkane containing 1-4 carbon atoms A linear or branched alkyl group derived from one or more hydrogen atoms, specifically, C _1-4 alkyl groups include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl base, sec-butyl, tert-butyl.

In the present invention, "C _1-6 alkoxy" refers to the above defined "C _1-6 alkyl" group attached through an oxygen atom to the remainder of the molecule, i.e., "C _1-6 alkyl -O -" groups, including but not limited to, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, neo- Pentyloxy, n-hexyloxy, etc.; the "C _1-4 alkoxy" refers to a group in which the above-defined "C _1-4 alkyl" is connected to the rest of the molecule through an oxygen atom, that is, " C _1-4 alkyl-O-" groups, including but not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy , tert-butoxy.

In the present invention, "halogenated C _1-6 alkyl" means that one or more (preferably 1-5, more preferably 1, 2 or 3) halogen atoms replace the above-defined "C _1-6 alkyl" A group formed by a corresponding number of hydrogen atoms on the "base", wherein each halogen atom can be the same or different when it is substituted with multiple halogen atoms. Specifically, the halogenated C _1-6 alkyl group includes but is not limited to monofluoromethyl group, trifluoromethyl, trichloromethyl, difluoromethyl, dichloromethyl, monochloromethyl, CF ₃ CH ₂ -, CHF ₂ CH ₂ -, CH ₂ FCH ₂ -, CH ₃ CHF-, CH ₂ FCHCl-, CF ₃ CH ₂ CH ₂ -, CF ₃ CH ₂ CH(CH ₃ )CH ₂ -, etc.; "halogenated C _1-4 alkyl" refers to one or more (preferably 1-5, More preferably 1, 2 or 3) halogen atoms replace _{the group formed by the corresponding number of hydrogen atoms on the "C 1-4} alkyl" as defined above, wherein when there are multiple halogen atoms substituted, each halogen atom Can be the same or different, specifically, including but not limited to monofluoromethyl, trifluoromethyl, trichloromethyl, difluoromethyl, dichloromethyl, monochloromethyl, CF ₃ CH ₂ -, CHF ₂ CH ₂ -, CH ₂ FCH ₂ -, CH ₃ CHF-, CH ₂ FCHCl- and the like.

In the present invention, "halogenated C _1-6 alkoxy" means that one or more (preferably 1-5, more preferably 1, 2 or 3) halogen atoms are substituted for the above-defined "C _{1-6 alkoxy group"} A group formed by the corresponding number of hydrogen atoms on "alkoxy", wherein each halogen atom can be the same or different when substituted by multiple halogen atoms, specifically, including but not limited to monofluoromethoxy, trifluoromethyl Oxy, trichloromethoxy, difluoromethoxy, dichloromethoxy, monochloromethoxy, CF ₃ CH ₂ -O-, CHF ₂ CH ₂ -O-, CH ₂ FCH ₂ -O- , CF ₃ CH ₂ CH ₂ -O-, etc.; "halogenated C _1-4 alkoxy" means that one or more (preferably 1-5, more preferably 1, 2 or 3) halogen atoms replace the above The group formed by the corresponding number of hydrogen atoms on the defined "C _1-4 alkoxy", wherein each halogen atom can be the same or different when it is substituted with multiple halogen atoms, specifically, including but not limited to a Fluoromethoxy, trifluoromethoxy, trichloromethoxy, difluoromethoxy, dichloromethoxy, monochloromethoxy, CF ₃ CH ₂ -O-, CHF ₂ CH ₂ -O- , CH ₂ FCH ₂ -O-, CF ₃ CH ₂ CH ₂ -O-, etc.

In the present invention, "hydroxy C _1-6 alkyl" means that one or more (preferably 1, 2 or 3) hydroxyl groups replace the corresponding number of hydrogen atoms on the _{above-defined "C 1-6 alkyl"} Formed groups, including but not limited to hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 2,3-dihydroxypropyl, 2-hydroxy -2-methylpropyl, 4-hydroxybutyl, 5-hydroxypentyl, 2-hydroxymethyl-1-hydroxypropyl; "hydroxy C _1-4 alkyl" means one or more (preferably 1 , 2 or 3) hydroxyl groups substituted _{for the corresponding number of hydrogen atoms on the "C 1-4} alkyl" defined above, including but not limited to hydroxymethyl, 1-hydroxyethyl, 2- Hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 2,3-dihydroxypropyl, 2-hydroxy-2-methylpropyl, 4-hydroxybutyl.

In the present invention, "ring atom" refers to the atoms forming a ring, including but not limited to C, N, O, P and S; "ring carbon atom" refers to the ring atom C; "ring heteroatom" refers to the atoms other than C atoms External ring atoms, including but not limited to ring atoms N, O, P and S.

In the present invention, "cycloalkyl" refers to a saturated or partially unsaturated cyclic hydrocarbon group, which can be a monovalent group or a divalent or higher group, including monocyclic cycloalkyl and polycyclic cycloalkyl, Polycyclic cycloalkyl includes spiro cycloalkyl, condensed cycloalkyl and bridged cycloalkyl, the "spirocycloalkyl" refers to the single ring sharing one ring carbon atom (called spiro atom). Polycyclic cycloalkyl, the "fused ring cycloalkyl" refers to a polycyclic cycloalkyl in which each ring in the group shares an adjacent pair of ring carbon atoms with other rings, and the "bridged cycloalkane" "Radical" refers to a polycyclic cycloalkyl group in which any two rings share two ring carbon atoms that are not directly attached. In the present invention, "3-6 membered cycloalkyl" refers to a cycloalkyl group including 3 to 6 ring carbon atoms, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclo Hexyl, cyclohexenyl, cyclohexadienyl, etc.

In the present invention, "heterocyclyl" refers to a saturated or partially unsaturated wherein one or more (preferably 1, 2, 3 or 4) ring atoms are ring heteroatoms selected from N, O or S cyclic hydrocarbon group, which can be a monovalent group or a divalent or more group, including monocyclic heterocyclic group and polycyclic heterocyclic group, polycyclic heterocyclic group including spirocyclic heterocyclic group, fused ring heterocyclic group and bridged ring heterocyclyl, the "spirocyclic heterocyclyl" refers to a polycyclic heterocyclyl group in which a single ring atom (called a spiro atom) is shared between the single rings, and the "fused ring cycloalkyl" refers to a radical A polycyclic heterocyclic group in which each ring in the group shares an adjacent pair of ring atoms with other rings, the "bridged ring cycloalkyl" refers to a polycyclic heterocyclic group in which any two rings share two ring atoms that are not directly connected. Cyclic heterocyclyl. In addition, the heterocyclic group also includes the case where the ring atoms C and S are oxo, for example, the ring atoms C and S are replaced by C(=O), S(=O), S(=O) ₂ . In the present invention, "3-8 membered heterocyclic group" refers to a heterocyclic group comprising 3 to 8 ring atoms, preferably containing 1, 2 or 3 ring heteroatoms selected from N, O or S, specifically , 3-8 membered heterocyclic groups include, but are not limited to, azetidinyl, oxetanyl, azetidinyl, oxetanyl, thietanyl, tetrahydrofuranyl, tetrahydropyrrolyl , pyrrolidone, tetrahydrothienyl, imidazolidinyl, pyrazolidine, 1,2-oxazolidinyl, 1,3-oxazolidinyl, 1,2-thiazolidinyl, 1,3-thiazolidine base, tetrahydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, piperidinyl, piperidonyl, piperazinyl, morpholinyl, 1,4-dioxanyl, 1 , 4-oxathiolanyl, 4,5-dihydroisoxazolyl, 4,5-dihydrooxazolyl, 2,5-dihydrooxazolyl, 2,3-dihydrooxazole base, 3,4-dihydro-2H-pyrrolyl, 2,3-dihydro-1H-pyrrolyl, 2,5-dihydro-1H-imidazolyl, 4,5-dihydro-1H-imidazolyl, 4,5-dihydro-1H-pyrazolyl, 4,5-dihydro-3H-pyrazolyl, 4,5-dihydrothiazolyl, 2,5-dihydrothiazolyl, 2H-pyranyl, 4H-pyranyl, 2H-thiopyranyl, 4H-thiopyranyl, 2,3,4,5-tetrahydropyridyl, 1,2-isoxazinyl, 1,4-isoxazinyl or 6H -1,3-oxazinyl etc., preferably

Wait. In the present invention, "3-6 membered heterocyclic group" refers to a heterocyclic group including 3 to 6 ring atoms, preferably containing 1, 2 or 3 ring heteroatoms selected from N, O or S, specifically , 3-6 membered heterocyclyl groups include but are not limited to azetidinyl, oxetanyl, azetidinyl, oxetanyl, thietanyl, tetrahydrofuranyl, tetrahydropyrrolyl , pyrrolidone, tetrahydrothienyl, imidazolidinyl, pyrazolidine, 1,2-oxazolidinyl, 1,3-oxazolidinyl, 1,2-thiazolidinyl, 1,3-thiazolidine base, tetrahydro-2H-pyranyl, tetrahydro-2H-thiopyranyl, piperidinyl, piperidonyl, piperazinyl, morpholinyl, 1,4-dioxanyl, 1 , 4-oxathiolanyl, 4,5-dihydroisoxazolyl, 4,5-dihydrooxazolyl, 2,5-dihydrooxazolyl, 2,3-dihydrooxazole base, 3,4-dihydro-2H-pyrrolyl, 2,3-dihydro-1H-pyrrolyl, 2,5-dihydro-1H-imidazolyl, 4,5-dihydro-1H-imidazolyl, 4,5-dihydro-1H-pyrazolyl, 4,5-dihydro-3H-pyrazolyl, 4,5-dihydrothiazolyl, 2,5-dihydrothiazolyl, 2H-pyranyl, 4H-pyranyl, 2H-thiopyranyl, 4H-thiopyranyl, 2,3,4,5-tetrahydropyridyl, 1,2-isoxazinyl, 1,4-isoxazinyl or 6H -1,3-oxazinyl etc., preferably

Wait.

In the present invention, "aryl" refers to an aromatic cyclic hydrocarbon group, which can be a monovalent group or a group with more than two valences, including monocyclic aryl groups and fused-ring aryl groups, "fused-ring aryl groups" Refers to an aryl group containing multiple rings (preferably containing 2 or 3 rings) in which each ring in the group shares an adjacent pair of ring carbon atoms with other rings. In the present invention, "6-10 membered aryl group" refers to an aryl group including 6 to 10 ring carbon atoms, including phenyl and naphthyl.

In the present invention, "heteroaryl" refers to a cyclic heterocyclic group in which one or more (preferably 1-5, more preferably 1, 2, 3 or 4) ring atoms are selected from N, O or S. Atomic aromatic cyclic hydrocarbon group, which can be a monovalent group or a divalent or higher group, including a monocyclic heteroaryl group and a fused ring heteroaryl group, the "fused ring heteroaryl group" refers to the radical A heteroaryl group containing multiple rings (preferably containing 2 or 3 rings) in which each ring in the group shares an adjacent pair of ring atoms with the other rings. In the present invention, "5-12-membered heteroaryl group" refers to a heteroaryl group including 5 to 12 ring atoms, preferably containing 1, 2, 3 or 4 heterocyclic rings selected from N, O or S Atoms, specific, 5-12 membered heteroaryl groups include, but are not limited to, furanyl, imidazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, oxazolyl, pyridyl, pyridazinyl, Pyrimidyl, pyrazinyl, pyrazolyl, pyrrolyl, tetrazolyl, thiadiazolyl, thienyl, triazolyl, triazinyl, benzimidazolyl, benzofuranyl, benzothienyl, benzene oxadiazolyl, benzothiadiazolyl, benzothiazolyl, furopyridyl, indazolyl, indolyl, isoindolyl, isoquinolinyl, purinyl, quinolinyl, quinoxa olinyl, thienopyridyl, etc., preferably

Wait. In the present invention, "5-6 membered heteroaryl" refers to a heteroaryl group comprising 5 to 6 ring atoms, preferably containing 1, 2, 3 or 4 ring heteroaryls selected from N, O or S Atoms, specific, 5-6 membered heteroaryl groups include but are not limited to furyl, imidazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, oxazolyl, pyridyl, pyridazinyl, Pyrimidyl, pyrazinyl, pyrazolyl, pyrrolyl, tetrazolyl, thiadiazolyl, thienyl, triazolyl, triazinyl, etc., preferably

Wait.

In the present invention, "-C(=O)-C _1-4 alkyl" refers to a group formed by connecting the above-defined "C _1-4 alkyl" and -C(=O)-; " -SO ₂ -C _1-4 alkyl" refers to a group formed by attaching the above-defined "C _1-4 alkyl" to -SO _{2 -.}

In the present invention, the "more than one" means that the number of substituents can be the number of all chemically substituted positions of the substituted group, preferably 1-6, more preferably 1-5, more preferably 1 -3, more preferably 1-2, more preferably 1.

In the present invention, "amino protecting group" refers to a chemical group attached to an amino group and easily removed under certain conditions, including but not limited to alkoxycarbonyl, acyl, and alkyl; for example, tert-butoxycarbonyl , benzyloxycarbonyl, fluorenemethoxycarbonyl, allyloxycarbonyl, phthaloyl, benzyl, p-methoxybenzyl, trityl, etc. Those skilled in the art can refer to Greene's Protective Groups in Organic Synthesis (4 ^th edition), a textbook commonly used in the art, for appropriate selection and operation.

The pharmaceutical composition of the present invention can be prepared by methods well known in the art, such as conventional mixing method, dissolving method, granulation method, sugar-coated pill method, grinding method, emulsification method and freeze-drying method. The pharmaceutical compositions of the present invention can be administered to a subject by any convenient route of administration, including, but not limited to, oral, rectal, parenteral (eg, injection, including subcutaneous, intradermal, intramuscular, intravenous, etc.), Topical (including, for example, transdermal, intranasal, ocular, buccal, and sublingual), pulmonary (eg, using aerosol for oral or nasal inhalation or insufflation therapy), and the like.

Solid dosage forms suitable for oral administration include tablets, pills, capsules, powders, powders, granules and the like. When preparing these solid dosage forms, in addition to the compounds described in the present invention or their pharmaceutically acceptable salts, excipients, fillers or compatibilizers, binders, disintegrants, stabilizers commonly used in the art can be added , one or more of wetting agents, adsorbents, lubricants or encapsulating materials.

Liquid dosage forms suitable for oral administration include solutions, suspensions, emulsions, syrups or tinctures and the like. In addition to the compounds described in the present invention or their pharmaceutically acceptable salts, liquid dosage forms may contain diluents, solubilizers, emulsifiers, wetting agents, suspending agents, sweeteners, flavoring agents, conventionally used in the art, One or more of fragrances or preservatives.

Dosage forms suitable for topical administration include ointments, powders, patches, drops, sprays, inhalants, etc., as the active ingredient of the compound of the present invention or a pharmaceutically acceptable salt thereof under sterile conditions and pharmaceutically with an acceptable carrier.

Dosage forms suitable for rectal administration include suppositories comprising a compound of the present invention or a pharmaceutically acceptable salt thereof together with a suitable base.

Dosage forms suitable for parenteral injection include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions.

The pharmaceutical formulations are preferably in unit dosage form. In this form, the preparation is subdivided into unit doses containing appropriate quantities of the active component. Unit dosage forms can be packaged in discrete quantities of preparation, such as packeted tablets, capsules.

In treating a patient, an effective amount of a compound of the present invention or a pharmaceutically acceptable salt or pharmaceutical composition thereof is administered to a patient (eg, a human) in need of treatment. In some embodiments, a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, is about 0.01 to 1000 mg/day, preferably about 0.1 to 500 mg/day. The precise dosage will depend upon the route of administration, the form in which the compound is to be administered, the health status of the subject (patient) to be treated and the experience of the attending physician, among others.

The compounds of the present invention or their pharmaceutically acceptable salts or their pharmaceutical compositions can also be administered in combination with other antineoplastic drugs, such as chemotherapeutic drugs, or in combination with other treatments such as radiation therapy or surgery, as a preoperative or adjuvant after surgery.

Abbreviations used in the present invention have the following meanings:

Boc: tert-butoxycarbonyl

Cbz: benzyloxycarbonyl

Bn: benzyl

Pd(dppf)Cl ₂ : [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride

Pd ₂ (dba) ₃ : tris(dibenzylideneacetone)dipalladium

Xantphos: 4,5-Bisdiphenylphosphine-9,9-dimethylxanthene

Brettphos: dicyclohexyl[3,6-dimethoxy-2',4',6'-triisopropyl[1,1'-biphenyl]-2-yl]phosphine

PtO ₂ : platinum dioxide

Pd(PPh ₃ ) ₄ : tetrakis(triphenylphosphine)palladium

PdCl ₂ : Palladium dichloride

Pd(OH) ₂ : palladium hydroxide

Pd(OAc) ₂ : palladium acetate

Pd: Palladium

HCl: hydrogen chloride

HBr: hydrogen bromide

NaBH(OAc) ₃ : sodium triacetoxyborohydride

NaBH ₄ : sodium borohydride

NaBH ₃ CN: Sodium cyanoborohydride

DCM: dichloromethane

MeOH: methanol

EA: Ethyl acetate

CDCl ₃ : deuterated chloroform

DMSO: Dimethyl Sulfoxide

TLC: Thin Layer Chromatography

Description of drawings:

Figure 1 shows the tumor volume change curve of the subcutaneously transplanted tumor of Ba/F3 KIF5B-RET nude mice in stable transgenic cells of the compound of Example 12 and LOXO-292 at a dose of 10 mg/kg twice a day.

Fig. 2 is the tumor volume change curve of the subcutaneously transplanted tumor of stably transduced cells Ba/F3 KIF5B-RET-G810R nude mice with the compound of Example 12 and LOXO-292 at a dose of 30 mg/kg twice a day.

detailed description

The solution of the present invention will be explained below with reference to specific embodiments. It should be understood that these examples are only intended to illustrate the present invention and not to limit the scope of the present invention. If no specific technique or condition is indicated in the following examples, the technique or condition described in the literature in this field or the product specification is used. The reagents or instruments used without the manufacturer's indication are conventional products that can be obtained from the market.

I. Preparation Examples of Compounds of the Invention

The structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) or/and liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts ([delta]) are given in parts per million (ppm). The measurement of NMR was carried out with a Bruker AVANCE-400 nuclear magnetic instrument, the solvents were deuterated dimethyl sulfoxide (DMSO-d ₆ ) and deuterated chloroform (CDCl ₃ ), and the internal standard was tetramethylsilane (TMS).

Agilent 6410 Triple Quad LC/MS was used for LC-MS determination.

The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the specifications used for TLC are 0.15mm-0.20mm, and the specifications used for TLC separation and purification products are 0.4mm-0.5mm. Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

Example 1: 6-(2,2-Difluoroethylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo [3.1.1]Heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

Step 1: 6-Bromo-4-(6-fluoropyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

Into a 500ml there-necked flask was added 4,6-dibromopyrazo[1,5-a]pyridine-3-carbonitrile (33.0g, 0.11mol), 2-fluoro-5-pyridineboronic acid (14.1g, 0.10mmol) , [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (2.7g, 3.70mmol), potassium fluoride dihydrate (31.0g, 0.33mmol) and 150ml N,N-dihydrate Methylformamide was reacted at 60°C overnight under the protection of argon. After the reaction was completed, it was cooled to room temperature, and 500 ml of water was added. The reaction solution was filtered, and the filter cake was rinsed with acetonitrile and ethyl acetate. Dry under reduced pressure to obtain 30 g of 6-bromo-4-(6-fluoropyridin-3-yl)pyrazo[1,5-a]pyridine-3-carbonitrile. The yield was 86%.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.48 (d, J=1.6 Hz, 1H), 8.73 (s, 1H), 8.51 (d, J=2.5 Hz, 1H), 8.27 (td, J= 8.1, 2.6Hz, 1H), 7.86 (d, J=1.6Hz, 1H), 7.40 (dd, J=8.4, 2.8Hz, 1H).

Step 2: 3-(5-(6-Bromo-3-cyanopyrazo[1,5-a]pyridin-4-yl)pyridin-2-yl)-3,6-diazabicyclo[3.1. 1] Heptane-6-carbamic acid tert-butyl ester

To a 250ml there-necked flask was added 6-bromo-4-(6-fluoropyridin-3-yl)pyrazo[1,5-a]pyridine-3-carbonitrile (4.35g, 13.7mmol), 6-(tert-butyl) oxycarbonyl)-3,6-diazabicyclo[3.1.1]heptane (3.25 g, 16.3 mmol), potassium carbonate (5.67 g, 41.1 mmol) and 60 ml of dimethyl sulfoxide, react at 100°C overnight. After the reaction was completed, it was cooled to room temperature, and 200 ml of water was added. The reaction solution was filtered, the filter cake was dissolved in a mixed solvent (DCM:MeOH=10:1) and then drained, acetonitrile was watered three times, and 3.4g of 3-(5-(6-bromo-3-cyanopyridine was obtained by silica gel column chromatography) oxazo[1,5-a]pyridin-4-yl)pyridin-2-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carbamic acid tert-butyl ester. The yield was 50%.

MS m/z: 495.1/497.1 [M+1] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.68 (d, J=1.3 Hz, 1H), 8.36 (d, J=2.1 Hz, 1H), 8.25 (s, 1H), 7.72 (dd, J=8.8, 2.3Hz, 1H), 7.39 (d, J=1.3Hz, 1H), 6.66 (d, J=8.8Hz, 1H), 4.31 (d, J=4.3Hz, 2H), 4.15 (s, 2H), 3.55 (s, 2H), 2.68 (d, J=7.6Hz, 1H), 1.53 (d, J=8.7Hz, 1H), 1.37 (d, J=5.2Hz, 9H).

Step 3: 4-(6-(3,6-Diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)-6-bromopyrazolo[1,5-a]pyridine- 3-carbonitrile

Add 3-(5-(6-bromo-3-cyanopyrazo[1,5-a]pyridin-4-yl)pyridin-2-yl)-3,6-diazabicyclo to a 250ml there-necked flask [3.1.1] Heptane-6-carbamic acid tert-butyl ester (3.4 g, 6.88 mmol) and 50 ml of 4.0 M methanol solution of hydrogen chloride, react at 25°C overnight. The MS monitoring has been reflected. The reaction solution was drained and water was added to acetonitrile three times to obtain 3.66g of 4-(6-(3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)-6-bromo Crude pyrazo[1,5-a]pyridine-3-carbonitrile. The yield was 99%.

MS m/z: 395.0/397.0 [M+1] ⁺ .

Step 4: 6-Bromo-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3- yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

Add 4-(6-(3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)-6-bromopyrazo[1,5-a to a 250ml there-necked flask ] pyridine-3-carbonitrile crude product (3.66g, 6.88mmol), 6-methoxy-3-pyridinecarbaldehyde (1.88g, 13.76mmol) and 100ml of dichloroethane, slowly added sodium triacetylborohydride (7.3 g, 34.4 mmol), reacted overnight at room temperature, and TLC monitored the completion of the reaction. Saturated sodium bicarbonate quenched the reaction, washed with water, extracted with dichloromethane, dried the organic phase, sucked dry, and chromatographed on silica gel column to obtain 2.8g of 6-bromo-4-(6-(6-((6-methoxypyridine- 3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile. The yield was 79%.

MS m/z: 516.1/518.1 [M+1] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.70 (d, J=1.5 Hz, 1H), 8.40 (d, J=2.2 Hz, 1H), 8.26 (s, 1H), 8.11 (d, J=2.0 Hz) , 1H), 7.77 (dd, J=8.8, 2.5Hz, 2H), 7.40 (d, J=1.5Hz, 1H), 6.71 (dd, J=15.8, 8.7Hz, 2H), 3.90 (d, J= 12.7Hz, 7H), 3.68 (d, J=9.2Hz, 4H), 2.84 (s, 1H), 1.71 (d, J=9.0Hz, 1H).

Step 5: 6-(2,2-Difluoroethylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[ 3.1.1]Heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

Add 6-bromo-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane to a 50ml single-necked flask -3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile (103 mg, 0.20 mmol), tris(dibenzylideneacetone)dipalladium (18.3 mg, 0.02 mmol) , dicyclohexyl[3,6-dimethoxy-2',4',6'-triisopropyl[1,1'-biphenyl]-2-yl]phosphine (10.8 mg, 0.02 mmol), Cesium carbonate (163 mg, 0.50 mmol), 2,2-difluoroethylamine (33 mg, 0.40 mmol) and 10 ml of dioxane were reacted at 90°C overnight under argon protection. Celite was used for filtration, the filtrate was evaporated to dryness under reduced pressure, and silica gel column chromatography (dichloromethane: methanol = 15:1) was used to obtain 77 mg of product 6-(2,2-difluoroethylamine)-4-(6-( 6-((6-Methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazol[1, 5-a]pyridine-3-carbonitrile. The yield was 74%.

MS m/z: 517.2 [M+1] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.44(s, 1H), 8.36(s, 1H), 8.25(s, 1H), 8.08(s, 1H), 7.80(d, J=8.0Hz, 1H), 7.69(d, J=8.0Hz, 1H), 7.15(s, 1H), 6.79(t, J=8.0Hz, 2H), 6.32(t, J=8.0Hz, 1H), 6.22(tt, J ₁ =56.0Hz, J ₂ =4.0Hz, 1H), 3.83(s, 3H), 3.79-3.61(m, 4H), 3.59-3.46(m, 4H), 3.39-3.28(m, 2H), 2.50 (s, 1H), 1.60 (d, J=8.0 Hz, 1H).

Example 2: 6-(Ethylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane Alk-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. Using intermediate 2A and ethylamine as raw materials, the target compound 6-(ethylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-di Azabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile. The yield was 30%.

MS m/z: 481.5 [M+1] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.39(s, 1H), 8.34(s, 1H), 8.09(s, 1H), 7.93(s, 1H), 7.78(d, J=8.7Hz, 1H), 7.71(s, 1H), 7.07(s, 1H), 6.90-6.67(m, 2H), 6.01(s, 1H), 3.82(s, 3H), 3.72(s, 4H), 3.53(s , 4H), 3.14-2.98(m, 2H), 2.07(s, 1H), 1.59(s, 1H), 1.22(t, J=6.9Hz, 3H).

Example 3: 4-(6-(6-((6-Methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine -3-yl)-6-(n-propylamine)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. Using intermediate 2A and n-propylamine as raw materials, the target compound 4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1] was obtained Heptan-3-yl)pyridin-3-yl)-6-(n-propylamine)pyrazolo[1,5-a]pyridine-3-carbonitrile. The yield was 13%.

MS m/z: 495.3 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.40 (s, 1H), 8.35 (d, J=2.0 Hz, 1H), 8.09 (s, 1H), 7.95 (d, J=2.0 Hz, 1H) , 7.80(d, J=8.0Hz, 1H), 7.70(d, J=8.0Hz, 1H), 7.07(d, J=2.0Hz, 1H), 6.85-6.74(m, 2H), 5.98(t, J=8.0Hz, 1H), 3.83(s, 3H), 3.80-3.63(m, 4H), 3.62-3.45(m, 4H), 3.10-2.98(m, 2H), 2.50(s, 1H), 1.68 -1.54 (m, 3H), 0.99 (t, J=6.0 Hz, 3H).

Example 4: 4-(6-(6-((6-Methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine -3-yl)-6-(2,2,2-trifluoroethylamine)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. Using intermediate 2A and trifluoroethylamine as raw materials, the target compound 4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1 .1]Heptan-3-yl)pyridin-3-yl)-6-(2,2,2-trifluoroethylamine)pyrazo[1,5-a]pyridine-3-carbonitrile. The yield was 68%.

MS m/z: 535.4 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.46 (s, 1H), 8.37 (d, J=2.0 Hz, 2H), 8.09 (s, 1H), 7.81 (d, J=8.0 Hz, 1H) , 7.74-7.66(m, 1H), 7.18(d, J=2.0Hz, 1H), 6.86-6.74(m, 2H), 6.60(t, J=8.0Hz, 1H), 4.18-4.05(m, 2H) ), 3.83 (s, 3H), 3.80-3.63 (m, 4H), 3.62-3.40 (m, 4H), 2.50 (s, 1H), 1.60 (s, 1H).

Example 5: 6-(isobutylamino)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1 ]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. Using intermediate 2A and isobutylamine as raw materials, the target compound 6-(isobutylamino)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3 was obtained, 6-Diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile. The yield was 60%.

MS m/z: 509.3 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.40 (s, 1H), 8.36 (s, 1H), 8.09 (s, 1H), 7.94 (s, 1H), 7.80 (d, J=8.0 Hz, 1H), 7.70(d, J=8.0Hz, 1H), 7.11(s, 1H), 6.85-6.75(m, 2H), 6.00(t, J=8.0Hz, 1H), 3.83(s, 3H), 3.79-3.63(m, 4H), 3.62-3.42(m, 4H), 2.89(t, J=6.0Hz, 2H), 1.96-1.85(m, 1H), 2.50(s, 1H), 1.60(s, 1H), 0.99 (t, J=6.4Hz, 6H).

Example 6: 6-(3,3-Difluorocyclobutylamino)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazepine Heterobicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. Using intermediate 2A and 3,3-difluorocyclobutylamine as raw materials, the target compound 6-(3,3-difluorocyclobutylamino)-4-(6-(6-((6-methoxy Pyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-methyl Nitrile. The yield was 50%.

MS m/z: 543.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.45 (s, 1H), 8.36 (d, J=1.88 Hz, 1H), 8.09 (br.s., 1H), 7.98 (s, 1H), 7.80 (d, J=7.52Hz, 1H), 7.70 (d, J=5.91Hz, 1H), 7.02 (s, 1H), 6.73-6.86 (m, 2H), 6.56 (d, J=6.18Hz, 1H) , 3.88-3.86(m, 1H), 3.83(s, 3H), 3.76-3.64(m, 4H), 3.60-3.45(m, 4H), 3.17(tt, J=7.29, 13.94Hz, 2H), 2.61 -2.49(m, 3H), 1.60(s, 1H).

Example 7: 6-(Cyanomethylamino)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1 ]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. Using intermediate 2A and aminoacetonitrile as raw materials, the target compound 6-(cyanomethylamino)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6 was obtained - Diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile. The yield was 15%.

MS m/z: 492.2[M+1] ⁺ .

Example 8: 6-(2-Hydroxyethylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1. 1]Heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. Using intermediate 2A and aminoethanol as raw materials, the target compound 6-(2-hydroxyethylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3 was obtained, 6-Diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile. The yield was 15%.

MS m/z: 497.4 [M+1] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.41(s, 1H), 8.35(s, 1H), 8.10(s, 1H), 8.02(s, 1H), 7.80(d, J=8.0Hz, 1H), 7.70(d, J=8.0Hz, 1H), 7.15(s, 1H), 6.86-6.73(m, 2H), 5.99(t, J=8.0Hz, 1H), 4.78(t, J=5.2 Hz, 1H), 3.96-3.85(m, 2H), 3.83(s, 3H), 3.79-3.63(m, 4H), 3.61-3.44(m, 4H), 3.30(s, 2H), 3.21-3.10( m, 2H), 2.50 (s, 1H), 1.60 (s, 1H).

Example 9: 6-(2-Hydroxy-2-methylpropylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazepine Bicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. Using intermediate 2A and 1-amino 2-methyl 2-propanol as raw materials, the target compound 6-(2-hydroxy-2-methylpropylamine)-4-(6-(6-((6-methoxy pyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3- formonitrile. The yield was 8%.

MS m/z: 525.3 [M+1] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.39(s, 1H), 8.36(s, 1H), 8.10(s, 1H), 8.05(s, 1H), 7.80(d, J=8.0Hz, 1H), 7.70(d, J=8.0Hz, 1H), 7.26(s, 1H), 6.84-6.74(m, 2H), 5.83(t, J=8.0Hz, 1H), 4.56(s, 1H), 3.82(s, 3H), 3.78-3.66(m, 4H), 3.59-3.42(m, 4H), 3.00(d, J=6.0Hz, 2H), 2.50(s, 1H), 1.60(s, 1H) , 1.21 (s, 6H).

Example 10: 6-((3-Fluoroazetidin-3-yl)methylamino)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)- 3,6-Diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

Step 1: 3-((3-cyano-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1] Heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridin-6-ylamino)methyl)-3-fluoroazetidine-1-carboxylate tert-butyl ester

The synthesis of this compound is the same as that of Example 1. Using intermediate 2A and tert-butyl 3-(aminomethyl)-3-fluoroazetidine-1-carboxylate as raw materials, the target compound 3-((3-cyano-4-(6-( 6-((6-Methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazol[1, 5-a]Pyridin-6-ylamino)methyl)-3-fluoroazetidine-1-carboxylate tert-butyl ester. The yield was 74%.

MS m/z: 640.2[M+1] ⁺ .

Step 2: 6-((3-Fluoroazetidin-3-yl)methylamino)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3 , 6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

Add 3-((3-cyano-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1 .1]Heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridin-6-ylamino)methyl)-3-fluoroazetidine-1-carboxylic acid tert. Butyl ester (95 mg, 0.15 mmol), 4 ml of dichloromethane and 4 ml of trifluoroacetic acid were reacted at 25°C for 2 hours under the protection of argon. The reaction solution was evaporated to dryness under reduced pressure, 20 ml of dichloromethane and 10 ml of saturated sodium bicarbonate were added, the mixture was stirred until no bubbles were present, and the organic layer was separated, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Silica gel column chromatography (dichloromethane:methanol=15:1) to obtain 36 mg of product 6-((3-fluoroazetidin-3-yl)methylamino)-4-(6-(6-(( 6-Methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazo[1,5-a] Pyridine-3-carbonitrile. The yield was 44%.

MS m/z: 540.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.43 (s, 1H), 8.35 (d, J=2.2 Hz, 1H), 8.26 (s, 1H), 8.14 (s, 1H), 7.79 (dd, J=8.8, 2.4Hz, 1H), 7.74 (dd, J=5.5, 3.5Hz, 1H), 7.20 (s, 1H), 6.83-6.75 (m, 2H), 6.53 (t, J=5.9Hz, 1H) ), 4.18(d, J=2.6Hz, 1H), 4.13(s, 1H), 3.83(s, 3H), 3.82-3.79(m, 4H), 3.76-3.73(m, 4H), 3.64-3.60( m, 5H), 2.64 (d, J=18.9Hz, 1H), 1.65 (s, 1H).

Example 11: 4-(6-(6-(3-Fluoro-4-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine-3- yl)-6-((3-fluoroazetidin-3-yl)methylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

Step 1: 4-(6-(6-(3-Fluoro-4-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl) -6-Bromopyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of intermediate 2A. After deprotection of intermediate 1, reductive amination with 3-fluoro-4-methoxybenzaldehyde affords intermediate 2B. The yield was 44%.

MS m/z: 533.0 [M+1] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.69 (d, J=1.5 Hz, 1H), 8.40 (d, J=2.1 Hz, 1H), 8.27 (s, 1H), 7.76 (dd, J=8.8, 2.5Hz, 1H), 7.40 (d, J=1.5Hz, 1H), 7.15 (dd, J=12.2, 1.7Hz, 1H), 7.06 (d, J=8.4Hz, 1H), 6.89 (t, J= 8.5Hz, 1H), 6.69 (d, J=8.8Hz, 1H), 3.87 (s, 3H), 3.83 (d, J=6.1Hz, 4H), 3.59 (d, J=8.8Hz, 4H), 2.74 (d, J=6.9Hz, 1H), 1.66 (d, J=8.7Hz, 1H).

Step 2: 3-((4-(6-(6-(3-Fluoro-4-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine -3-yl)-3-cyanopyrazo[1,5-a]pyridin-6-ylamino)methyl)-3-fluoroazetidine-1-carboxylate tert-butyl ester

The synthesis of this compound is the same as that of step 1 of Example 10. The yield was 34%.

MS m/z: 657.3 [M+1] ⁺ .

Step 3: 4-(6-(6-(3-Fluoro-4-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl )-6-((3-fluoroazetidin-3-yl)methylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of step 2 of Example 10. The yield was 30%.

MS m/z: 557.0 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.43 (s, 1H), 8.35 (d, J=2.2 Hz, 1H), 8.26 (d, J=1.2 Hz, 1H), 7.79 (dd, J= 8.8, 2.3Hz, 1H), 7.25-7.05 (m, 4H), 6.78 (d, J=8.8Hz, 1H), 6.53 (t, J=6.2Hz, 1H), 4.17-4.12 (m, 1H), 4.11(s, 1H), 3.81(s, 3H), 3.78-3.70(m, 6H), 3.65-3.48(m, 6H), 2.60(s, 1H), 1.62(d, J=7.8Hz, 1H) .

Example 12: 6-(2-Fluoroethylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1. 1]Heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

Step 1: 3-cyano-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3 -yl)pyridin-3-yl)pyrazolo[1,5-a]pyridin-6-ylcarbamate tert-butyl ester

The synthesis of this compound is the same as that of step 5 of Example 1. It was synthesized from intermediate 2A and tert-butyl carbamate as raw materials. The yield was 68%.

MS m/z: 553.4 [M+1] ⁺ .

Step 2: 3-cyano-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3 -yl)pyridin-3-yl)pyrazolo[1,5-a]pyridin-6-yl(2-fluoroethyl)carbamate tert-butyl ester

Add 3-cyano-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane to a 25ml single-necked flask Alk-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridin-6-ylcarbamate tert-butyl ester (55 mg, 0.10 mmol), cesium carbonate (66 mg, 0.20 mmol), 1-fluoro -2-Iodoethane (35 mg, 0.20 mmol) and 5 ml of DMF were reacted at 80°C for 4 hours under nitrogen protection. The reaction solution was quenched by adding 15 ml of water, and extracted three times with ethyl acetate (15 ml). The organic layer was evaporated to dryness under reduced pressure, and the crude product was subjected to silica gel column chromatography (dichloromethane:methanol=30:1) to obtain 60 mg of the product 3-cyano-4-(6-(6-((6-methoxypyridine-3). -yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridin-6-yl(2- Fluoroethyl) tert-butyl carbamate. The yield was 99%.

MS m/z: 599.5 [M+1] ⁺ .

Step 3: 6-(2-Fluoroethylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1 ]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

Add 3-cyano-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane to a 10ml single-necked flask Alk-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridin-6-yl(2-fluoroethyl)carbamate (60 mg, 0.10 mmol) and 5 ml of 4.0 M hydrogen chloride in methanol The solution was reacted at 25°C for 2 hours under the protection of argon. The reaction solution was evaporated to dryness under reduced pressure, 20 ml of dichloromethane and 10 ml of saturated sodium bicarbonate were added, and the mixture was stirred until there were no bubbles, and the organic layer was separated, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Silica gel column chromatography (dichloromethane : methanol = 15 : 1) to obtain 20 mg of the product 6-(2-fluoroethylamine)-4-(6-(6-((6-methoxypyridin-3-yl) Methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile. The yield was 40%.

MS m/z: 499.3 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.41 (s, 1H), 8.35 (d, J=1.6 Hz, 1H), 8.11 (d, J=1.4 Hz, 2H), 7.79 (d, J= 7.5Hz, 1H), 7.70(s, 1H), 7.12(s, 1H), 6.80(d, J=8.5Hz, 2H), 6.25(t, J=5.8Hz, 1H), 4.62(dt, J= 47.8, 4.7Hz, 2H), 3.83(s, 3H), 3.80-3.61(m, 4H), 3.60-3.50(m, 4H), 3.43(ddd, J=14.5, 11.1, 5.9Hz, 2H), 2.50 (s, 1H), 1.62(s, 1H).

Example 13: 4-(6-(6-(3-Fluoro-4-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine-3- yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

Step 1: 3-cyano-4-(6-(6-((3-fluoro-4-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl )pyridin-3-yl)pyrazolo[1,5-a]pyridin-6-ylcarbamate tert-butyl ester

The synthesis of this compound is the same as that of step 5 of Example 1. It was synthesized from intermediate 2B and tert-butyl carbamate as raw materials.

MS m/z: 570.2 [M+1] ⁺ .

Step 2: 3-Carbononitrile-4-(6-(6-((3-fluoro-4-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl )pyridin-3-yl)pyrazolo[1,5-a]pyridin-6-yl(2-fluoroethyl)carbamate tert-butyl ester

The synthesis of this compound is the same as that of step 2 of Example 12.

MS m/z: 616.2[M+1] ⁺ .

Step 3: 4-(6-(6-(3-Fluoro-4-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl )-6-(2-Fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of 6-(2-fluoroethylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1 .1] Heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile step. The yield was 28%.

MS m/z: 516.2 [M+1] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.42 (s, 1H), 8.35 (d, J=1.5Hz, 1H), 8.11 (d, J=1.5Hz, 1H), 7.88-7.64 (m, 2H), 7.13 (d, J=1.7Hz, 2H), 6.80 (d, J=8.8Hz, 1H), 6.27 (t, J=5.9Hz, 1H), 4.63 (dt, J=47.8, 4.8Hz, 2H), 3.83(s, 3H), 3.78-3.66(m, 4H), 3.63-3.50(m, 4H), 3.44(ddd, J=28.5, 10.1, 5.1Hz, 2H), 2.64-2.54(m, 1H), 1.63 (s, 1H).

Example 14: 4-(6-(6-(2-Fluoro-5-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine-3- yl)-6-(2,2-difluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

Step 1: 3-(5-(3-Cyano-6-(2,2-difluoroethylamino)pyrazolo[1,5-a]pyridin-4-yl)pyridin-2-yl)-3 , tert-butyl 6-diazabicyclo[3.1.1]heptane-6-carbamate

The synthesis of this compound is the same as that of step 5 of Example 1. It was synthesized from intermediates 1 and 2,2-difluoroethylamine. The yield was 61%.

MS m/z: 496.2 [M+1] ⁺ .

Step 2: 4-(6-(3,6-Diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)-6-(2,2-difluoroethylamino)pyridine Azole[1,5-a]pyridine-3-carbonitrile

Add 3-(5-(3-cyano-6-(2,2-difluoroethylamino)pyrazo[1,5-a]pyridin-4-yl)pyridin-2-yl to a 25ml there-necked flask )-3,6-diazabicyclo[3.1.1]heptane-6-carbamic acid tert-butyl ester (245mg, 0.50mmol) and 5ml of 4.0M methanol solution of hydrogen chloride, react at 25°C overnight. The MS monitoring has been reflected. The reaction solution was drained, and acetonitrile was added with water three times to obtain 300 mg of 4-(6-(3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)-6-(2, Crude 2-difluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile. The yield was 99%.

MS m/z: 396.2 [M+1] ⁺ .

Step 3: 4-(6-(6-(2-Fluoro-5-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl )-6-(2,2-difluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of step 4 of Example 1. Synthesized from intermediate 3A and 2-fluoro-5-methoxybenzaldehyde. The yield was 23%.

MS m/z: 534.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.43 (s, 1H), 8.35 (d, J=2.2, 1H), 8.24 (d, J=1.6, 1H), 7.79 (dd, J=8.7, 2.2, 1H), 7.18 (d, J=1.8, 1H), 7.06 (d, J=8.3, 2H), 6.81 (t, J=8.1, 2H), 6.41 (t, J=6.5, 1H), 6.22 (t, J=3.7, 1H), 3.75 (s, 3H), 3.75-3.54 (m, 11H), 2.60 (s, 1H), 1.63 (s, 1H).

Example 15: 4-(6-(6-(4-Methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)-6 -(2-Fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

Step 1: 3-(5-(6-((tert-butoxycarbonyl)amino)-3-cyanopyrazolo[1,5-a]pyridin-4-yl)pyridin-2-yl)-3,6 - tert-butyl diazabicyclo[3.1.1]heptane-6-carbamate

Add 3-(5-(6-bromo-3-cyanopyrazo[1,5-a]pyridin-4-yl)pyridin-2-yl)-3,6-diazabicyclo to a 100ml single-necked flask [3.1.1] Heptane-6-carbamic acid tert-butyl ester (635 mg, 1.28 mmol), tris(dibenzylideneacetone)dipalladium (118 mg, 0.13 mmol), dicyclohexyl[3,6-dimethoxy yl-2',4',6'-triisopropyl[1,1'-biphenyl]-2-yl]phosphine (69 mg, 0.13 mmol), cesium carbonate (1.26 g, 3.84 mmol), tert-carbamic acid Butyl ester (453 mg, 3.84 mmol) and 15 ml of dioxane were reacted at 90°C overnight under argon protection. Celite was used for filtration, the filtrate was evaporated to dryness under reduced pressure, and silica gel column chromatography (dichloromethane: methanol = 30:1) was used to obtain 488 mg of product 3-(5-(6-((tert-butoxycarbonyl)amino)-3 -Cyanopyrazo[1,5-a]pyridin-4-yl)pyridin-2-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carbamic acid tert-butyl ester. The yield was 71%.

MS m/z: 532.3 [M+1] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.12 (s, 1H), 8.36 (s, 1H), 8.23 (s, 1H), 7.76 (d, J=7.3, 1H), 7.10 (s, 1H), 6.67 (t, J=7.3, 2H), 4.32 (d, J=4.5, 2H), 4.23-4.07 (m, 2H), 3.59-3.47 (m, 2H), 2.69 (d, J=6.5, 1H) , 1.55(s, 9H), 1.43(s, 1H), 1.39(s, 9H).

Step 2: 3-(5-(6-((tert-butoxycarbonyl)(2-fluoroethyl)amino)-3-cyanopyrazo[1,5-a]pyridin-4-yl)pyridine-2 -yl)-3,6-diazabicyclo[3.1.1]heptane-6-carbamic acid tert-butyl ester

Add 3-(5-(6-((tert-butoxycarbonyl)amino)-3-cyanopyrazo[1,5-a]pyridin-4-yl)pyridin-2-yl)- 3,6-Diazabicyclo[3.1.1]heptane-6-carbamic acid tert-butyl ester (2900 mg, 5.46 mmol), cesium carbonate (5340 mg, 16.4 mmol), 1-fluoro-2-iodoethane (1430 mg) , 8.17mmol) and 100ml DMF, under nitrogen protection, 80 ℃ of reaction for 12 hours. The reaction solution was quenched by adding 150 ml of water, and extracted three times with ethyl acetate (150 ml). The organic layer was evaporated to dryness under reduced pressure, and the crude product was subjected to silica gel column chromatography (dichloromethane:methanol=30:1) to obtain 2820 mg of product. The yield was 89%.

MS m/z: 578.3 [M+1] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.57 (s, 1H), 8.37 (d, J=1.8, 1H), 8.28 (s, 1H), 7.76 (d, J=8.2, 1H), 7.30 (s , 1H), 6.68 (d, J=8.7, 1H), 4.69 (dt, J=47.5, 4.5Hz, 2H), 4.32 (d, J=4.6, 2H), 4.12 (dd, J=14.3, 7.1, 2H), 3.94(dt, J=26.5, 4.6Hz, 2H), 3.57(s, 2H), 2.69(dd, J=13.7, 6.2, 1H), 1.53(s, 1H), 1.48(s, 9H), 1.39(s, 9H).

Step 3: 4-(6-(3,6-Diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)-6-(2-fluoroethylamino)pyrazo[1 , 5-a]pyridine-3-carbonitrile

Add 3-(5-(6-((tert-butoxycarbonyl)(2-fluoroethyl)amino)-3-cyanopyrazo[1,5-a]pyridin-4-yl) to a 250ml single-necked flask Pyridin-2-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carbamic acid tert-butyl ester (2820 mg, 4.88 mmol) and 50 ml of a 4.0 M solution of hydrogen chloride in ethyl acetate, under argon , 25 ℃ reaction for 2 hours. The reaction solution was evaporated to dryness under reduced pressure, 200 ml of dichloromethane and 100 ml of saturated sodium bicarbonate were added, and the mixture was stirred until there were no bubbles, and the organic layer was separated, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Silica gel column chromatography (dichloromethane:methanol=15:1) gave the product. The yield was 80%.

MS m/z: 378.2 [M+1] ⁺ .

Step 4: 4-(6-(6-(4-Methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)-6- (2-Fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

Add 4-(6-(3,6-diazabicyclo[3.1.1]heptane-3-yl)pyridin-3-yl)-6-(2-fluoroethylamino)pyridine into a 25ml three-necked flask oxazol[1,5-a]pyridine-3-cyano (60 mg, 0.16 mmol), 4-methoxybenzaldehyde (18 mg, 0.13 mmol) and 6 ml of dichloroethane were added slowly with sodium triacetylborohydride ( 140 mg, 0.66 mmol), the reaction was carried out at room temperature overnight, and the completion of the reaction was monitored by TLC. Saturated sodium bicarbonate quenched the reaction, washed with water, extracted with dichloromethane, dried the organic phase, sucked dry, and chromatographed on silica gel column to obtain 4-(6-(6-(4-methoxybenzyl)-3,6-dichloromethane) Azabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)-6-(2-fluoroethylamino)pyrazolo[1,5-a]pyridine-3-cyano.

MS m/z: 498.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.41 (s, 1H), 8.35 (d, J=1.9, 1H), 8.10 (d, J=1.2, 1H), 7.80 (d, J=7.5, 1H), 7.31(s, 2H), 7.14(d, J=1.4, 1H), 6.90(s, 2H), 6.80(d, J=8.8, 1H), 6.30(t, J=5.6, 1H), 4.62(dt, J=47.7, 4.7Hz, 1H), 4.16-3.86(m, 4H), 3.74(s, 3H), 3.68-3.44(m, 4H), 3.43-3.37(m, 2H), 2.61( s, 1H), 1.66(s, 1H).

Example 16: 4-(6-(6-((5-Fluoro-6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3 -yl)pyridin-3-yl)-6-(2-fluoroethylamine)-pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 5-fluoro-6-methoxynicotinaldehyde.

MS m/z: 517.1 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.41 (s, 1H), 8.34 (d, J=2.5 Hz, 1H), 8.10 (d, J=1.9 Hz, 1H), 7.93 (s, 1H) , 7.78 (dd, J=8.8, 2.5Hz, 1H), 7.64 (d, J=11.5Hz, 1H), 7.12 (d, J=2.0Hz, 1H), 6.78 (d, J=8.8Hz, 1H) , 6.27(t, J=5.9Hz, 1H), 4.63(dt, J=47.7, 4.6Hz, 2H), 3.92(s, 3H), 3.71(s, 4H), 3.54(s, 4H), 3.43( ddd, J=28.5, 9.9, 5.0Hz, 2H), 2.55 (s, 1H), 1.59 (d, J=8.0Hz, 1H).

Example 17: 4-(6-(6-(3,5-Difluoro-4-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine -3-yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 3,5-difluoro-4-methoxybenzaldehyde.

MS m/z: 534.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.40 (s, 1H), 8.32 (d, J=2.3, 1H), 8.08 (d, J=1.5, 1H), 7.77 (dd, J=8.8, 2.4, 1H), 7.41-7.34 (m, 1H), 7.12 (d, J=8.2, 2H), 6.77 (d, J=8.8, 1H), 6.27 (t, J=5.8, 1H), 4.62 (dt , J=47.7, 4.7Hz, 2H), 3.87 (s, 3H), 3.72-3.65 (m, 4H), 3.69-3.50 (m, 6H), 2.56 (dd, J=12.9, 6.6, 1H), 1.64 (s, 1H).

Example 18: 4-(6-(6-(2-Fluoro-4-methoxybenzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine-3- yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 2-fluoro-4-methoxybenzaldehyde. The yield was 37%.

MS m/z: 516.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.41 (s, 1H), 8.35 (d, J=2.2, 1H), 8.10 (d, J=1.7, 1H), 7.79 (dd, J=8.7, 1.9, 1H), 7.40 (s, 1H), 7.14 (d, J=1.7, 1H), 6.78 (t, J=8.3, 3H), 6.25 (t, J=5.8, 1H), 4.69 (t, J = 4.7, 1H), 4.57 (t, J=4.7, 1H), 3.73 (s, 3H), 3.73-3.37 (m, 10H), 2.59 (s, 1H), 1.62 (s, 1H).

Example 19: 4-(6-(6-(4-(difluoromethoxy)benzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine-3- yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 4-(difluoromethoxy)benzaldehyde. The yield was 24%.

MS m/z: 534.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.41 (s, 1H), 8.35 (d, J=2.1, 1H), 8.10 (d, J=1.5, 1H), 7.79 (d, J=8.5, 1H), 7.42 (s, 2H), 7.25-6.98 (m, 4H), 6.77 (t, J=11.7, 1H), 6.25 (t, J=5.8, 1H), 4.69 (t, J=4.7, 1H) ), 4.57(t, J=4.7, 1H), 3.82-3.37(m, 10H), 2.57(s, 1H), 1.63(s, 1H).

Example 20: 4-(6-(6-((6-(difluoromethoxy)pyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3 -yl)pyridin-3-yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 6-(difluoromethoxy)nicotinaldehyde. The yield was 29%.

MS m/z: 535.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.41 (s, 1H), 8.35 (d, J=2.2, 1H), 8.21 (s, 1H), 8.10 (d, J=1.6, 1H), 7.91 (d, J=7.8, 1H), 7.79 (dd, J=8.8, 2.4, 1H), 7.68 (s, 1H), 7.12 (d, J=1.7, 1H), 7.04 (d, J=8.4, 1H) ), 6.79(d, J=8.8, 1H), 6.25(t, J=5.8, 1H), 4.69(t, J=4.7, 1H), 4.57(t, J=4.7, 1H), 3.82-3.37( m, 10H), 2.55 (s, 1H), 1.61 (d, J=7.7, 1H).

Example 21: 4-(6-(6-((3-Fluoro-4-(difluoromethoxy)benzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl )pyridin-3-yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 3-fluoro-4-(difluoromethoxy)benzaldehyde. MS m/z: 552.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.42 (s, 1H), 8.34 (d, J=2.2, 1H), 8.10 (d, J=1.5, 1H), 7.78 (dd, J=8.8, 2.4, 1H), 7.38 (d, J=12.1, 1H), 7.29 (t, J=8.2, 1H), 7.23 (d, J=11.5, 2H), 7.13 (d, J=1.6, 1H), 6.78 (d, J=8.8, 1H), 6.30 (t, J=5.8, 1H), 4.63 (dt, J=47.7, 4.7Hz, 2H), 3.72-3.68 (m, 4H), 3.58-3.53 (m, 4H), 3.43 (ddd, J=27.3, 9.5, 4.5Hz, 2H), 2.57 (dd, J=12.8, 6.6Hz, 1H), 1.60 (d, J=8.4, 1H).

Example 22: 4-(6-(6-((2,3-Difluoro-4-(methoxy)benzyl)-3,6-diazabicyclo[3.1.1]heptane-3- yl)pyridin-3-yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 2,3-difluoro-4-(methoxy)benzaldehyde.

MS m/z: 534.2 [M+1] ⁺ .

Example 23: 4-(6-(6-((2,5-Difluoro-4-(methoxy)benzyl)-3,6-diazabicyclo[3.1.1]heptane-3- yl)pyridin-3-yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 2,5-difluoro-4-(methoxy)benzaldehyde.

MS m/z: 534.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.41 (s, 1H), 8.34 (d, J=2.3, 1H), 8.10 (d, J=1.7, 1H), 7.78 (dd, J=8.8, 2.5, 1H), 7.28 (dd, J=12.0, 7.0, 1H), 7.13 (d, J=1.8, 1H), 7.06 (dd, J=11.5, 7.3, 1H), 6.77 (d, J=8.8, 1H), 6.27(t, J=5.8, 1H), 4.62(dt, J=47.7, 4.7Hz, 2H), 3.82(s, 3H), 3.74-3.68(m, 4H), 3.60-3.40(m, 6H), 2.53 (d, J=7.3, 1H), 1.58 (d, J=8.4, 1H).

Example 24: 4-(6-(6-((3-Chloro-4-(methoxy)benzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine -3-yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 3-chloro-4-(methoxy)benzaldehyde.

MS m/z: 532.2 [M+1] ⁺ , 534.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ=8.42 (s, 1H), 8.34 (d, J=2.3, 1H), 8.11 (d, J=1.6, 1H), 7.78 (dd, J=8.8, 2.4, 1H), 7.41 (s, 1H), 7.28 (d, J=8.2, 1H), 7.14 (s, 1H), 7.07 (d, J=8.5, 1H), 6.78 (d, J=8.8, 1H) ), 6.31(d, J=4.1, 1H), 4.63(dt, J=47.7, 4.7Hz, 2H), 3.83(s, 3H), 3.69(s, 4H), 3.56-3.45(m, 4H), 3.43-3.38 (m, 2H), 2.57 (s, 1H), 1.60 (d, J=8.1, 1H).

Example 25: 4-(6-(6-((2-Chloro-4-(methoxy)benzyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridine -3-yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 2-chloro-4-(methoxy)benzaldehyde.

MS m/z: 532.2 [M+1] ⁺ , 534.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.41 (s, 1H), 8.34 (d, J=2.2, 1H), 8.11 (d, J=1.6, 1H), 7.79 (dd, J=8.7, 2.3, 1H), 7.49 (s, 1H), 7.15 (d, J=1.0, 1H), 7.00 (s, 1H), 6.93 (d, J=7.7, 1H), 6.79 (d, J=8.8, 1H) ), 6.30(s, 1H), 4.63(dt, J=47.7, 4.7Hz, 2H), 3.83(s, 3H), 3.82-3.66(m, 4H), 3.61-3.51(m, 4H), 3.49- 3.38(m, 2H), 2.58(s, 1H), 1.62(s, 1H).

Example 26: 4-(6-(6-((5-Chloro-6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptane-3 -yl)pyridin-3-yl)-6-(2-fluoroethylamino)pyrazo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 15. It was synthesized from intermediate 3B and 5-chloro-6-methoxynicotinic aldehyde.

MS m/z: 532.2 [M+1] ⁺ , 534.2.

Example 27: 6-(4,4,4-Trifluorobutylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazepine Heterobicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 12. After the intermediate 2A is coupled with tert-butyl carbamate, it is nucleophilically substituted with 1,1,1-trifluoro-4-iodobutane, and then the protecting group is removed.

MS m/z: 563.2 [M+1] ⁺ .

¹ H NMR (400MHz, DMSO) δ 8.42(s, 1H), 8.36(d, J=2.3Hz, 1H), 8.09(s, 1H), 8.03(d, J=1.6Hz, 1H), 7.79( dd, J=8.8, 2.4Hz, 1H), 7.70 (d, J=7.8Hz, 1H), 7.05 (d, J=1.7Hz, 1H), 6.94-6.65 (m, 2H), 6.11 (t, J =5.6Hz, 1H), 3.83(s, 3H), 3.79-3.65(m, 4H), 3.55(d, J=18.8Hz, 4H), 3.16(dd, J=12.7, 6.5Hz, 3H), 2.52 (s, 1H), 2.48-2.33 (m, 3H), 1.92-1.74 (m, 2H), 1.60 (d, J=6.4Hz, 1H).

Example 28: 6-(3-Methanesulfonylpropylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1 .1]Heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. It was synthesized from intermediate 2A and 3-methanesulfonylpropylamine.

MS m/z: 573.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO) δ 8.42 (s, 1H), 8.35 (d, J=2.2 Hz, 1H), 8.07 (d, J=11.5 Hz, 2H), 7.79 (dd, J=8.8, 2.2 Hz, 1H), 7.69(d, J=7.7Hz, 1H), 7.04(s, 1H), 6.78(t, J=7.6Hz, 2H), 6.13(t, J=5.6Hz, 1H), 3.82( s, 3H), 3.78-3.65(m, 4H), 3.51(s, 4H), 3.31-3.18(m, 4H), 3.00(s, 3H), 2.52(s, 1H), 2.00(dd, J= 14.6, 7.1Hz, 2H), 1.59 (d, J=7.1Hz, 1H).

Example 29: 6-(3-Fluoropropylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1 ]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 12. After the intermediate 2A is coupled with tert-butyl carbamate, it is nucleophilically substituted with 1-fluoro-3-iodopropane, and then the protecting group is removed.

MS m/z: 513.2 [M+1] ⁺ .

Example 30: 6-(3,3,3-Trifluoropropylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6-diazepine Bicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. It was synthesized from intermediate 2A and 3,3,3-trifluoropropylamine as raw materials.

MS m/z: 549.2 [M+1] ⁺ .

Example 31: 6-(2,2,3,3,3-Pentafluoropropylamine)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3,6 - Diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. It was synthesized from intermediate 2A and 2,2,3,3,3-pentafluoropropylamine.

MS m/z: 585.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO) δ 8.46 (s, 1H), 8.35 (d, J=5.7 Hz, 2H), 8.08 (s, 1H), 7.80 (d, J=7.7 Hz, 1H), 7.69 ( d, J=8.1Hz, 1H), 7.20 (d, J=1.6Hz, 1H), 6.78 (t, J=9.4Hz, 2H), 6.53 (t, J=6.8Hz, 1H), 4.17 (td, J=15.8, 6.5Hz, 2H), 3.82 (s, 3H), 3.78-3.62 (m, 4H), 3.57-3.51 (m, 4H), 2.50 (s, 1H), 1.59 (d, J=8.0Hz) , 1H).

Example 32: 6-((2-(1H-imidazol-1-yl)ethylamino)-4-(6-(6-((6-methoxypyridin-3-yl)methyl)-3 , 6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. It was synthesized from intermediate 2A and 2-(1H-imidazol-1-yl)ethanamine.

MS m/z: 547.2 [M+1] ⁺ .

¹ H NMR (400 MHz, DMSO) δ 8.42 (s, 1H), 8.34 (d, J=1.9 Hz, 1H), 8.10 (s, 2H), 7.78 (dd, J=8.7, 2.2 Hz, 1H), 7.70(d, J=8.1Hz, 2H), 7.28(s, 1H), 7.07(s, 1H), 6.91(s, 1H), 6.78(dd, J=8.6, 4.2Hz, 2H), 6.20(t , J=5.7Hz, 1H), 4.20(t, J=5.7Hz, 2H), 3.83(s, 3H), 3.78-3.65(m, 4H), 3.63-3.51(m, 4H), 3.50-3.46( m, 2H), 2.56 (s, 1H), 1.61 (d, J=8.2Hz, 1H).

Example 33: 6-((2-(1H-1,2,4-triazol-1-yl)ethylamino)-4-(6-(6-((6-methoxypyridine-3- yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. It was synthesized from intermediate 2A and 2-(1H-1,2,4-triazol-1-yl)ethanamine.

MS m/z: 548.2 [M+1] ⁺ .

¹ H NMR (400MHz, DMSO) δ 8.54(s, 1H), 8.43(s, 1H), 8.35(d, J=2.2Hz, 1H), 8.11(s, 2H), 8.01(s, 1H), 7.78 (dd, J=8.7, 2.1Hz, 1H), 7.70 (d, J=7.9Hz, 1H), 7.05 (d, J=1.0Hz, 1H), 6.79 (dd, J=8.5, 3.3Hz, 2H) ), 6.17(t, J=5.9Hz, 1H), 4.42(t, J=5.7Hz, 2H), 3.83(s, 3H), 3.77-3.74(m, 4H), 3.60-3.56(m, 6H) , 2.55(s, 1H), 1.61(s, 1H).

Example 34: 6-((2-(1H-1,3,4-triazol-1-yl)ethylamino)-4-(6-(6-((6-methoxypyridine-3- yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as that of Example 1. It was synthesized from intermediate 2A and 2-(1H-1,3,4-triazol-1-yl)ethanamine.

MS m/z: 548.2 [M+1] ⁺ .

¹ H NMR (400MHz, DMSO) δ 8.55(s, 2H), 8.43(s, 1H), 8.35(d, J=2.3Hz, 1H), 8.17(d, J=1.7Hz, 1H), 8.08( s, 1H), 7.78 (dd, J=8.8, 2.5Hz, 1H), 7.68 (dd, J=8.4, 2.2Hz, 1H), 7.04 (d, J=1.8Hz, 1H), 6.86-6.72 (m , 2H), 6.17(t, J=6.0Hz, 1H), 4.26(t, J=5.8Hz, 2H), 3.82(s, 3H), 3.74-3.67(m, 4H), 3.57-3.51(m, 6H), 2.52(s, 1H), 1.59(d, J=8.3Hz, 1H).

Example 35: 6-((2-(4H-1,2,4-triazol-1-yl)ethylamino)-4-(6-(6-((6-methoxypyridine-3- yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carbonitrile

The synthesis of this compound is the same as in Example 12. After coupling of intermediate 2A and tert-butyl carbamate, with 3-(2-iodoethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-1,2, The 4-triazole is nucleophilically substituted and the protecting group is removed.

MS m/z: 548.2 [M+1] ⁺ .

Example 36: 6-((2-(5-Methyl-4H-1,2,4-triazol-3-yl)ethylamino)-4-(6-(6-((6-methoxy pyridin-3-yl)methyl)-3,6-diazabicyclo[3.1.1]heptan-3-yl)pyridin-3-yl)pyrazolo[1,5-a]pyridine-3- Formonitrile

The synthesis of this compound is the same as in Example 12. After coupling of intermediate 2A and tert-butyl carbamate, nucleophilic substitution with 3-(2-iodoethyl)-5-methyl-4H-1,2,4-triazole-4-carboxylic acid tert-butyl ester , the deprotecting group is obtained.

MS m/z: 562.2[M+1] ⁺ .

II. Biological Test Example

Test Example 1: Inhibitory activity against RET kinase

The inhibitory effect of compounds on RET kinase (purchased from Carna Biosciences, Inc., Cat. No. 08-159) was detected by the Caliper Mobility Shift Assay method based on the microfluidic chip technology developed by Caliper. The compounds of the invention, BLU-667 and LOXO-292 were tested at final concentrations of 2 μM or 10 μM starting at 10 concentrations in 3-fold dilutions. Use a dispenser Echo 550 to transfer 250 nL of each compound at a 100-fold final concentration into a 384-well reaction plate, add 10 μL of kinase solution (diluted RET kinase with buffer to a final concentration of 1 nM; buffer purchased from Sandia Medical Technologies) (Shanghai) Co., Ltd.), pre-incubated at room temperature for 10 minutes (negative control wells contained 10 μL buffer and 250 nL 100% DMSO; positive control wells contained 10 μL kinase solution and 250 nL 100% DMSO). Add 15 μL of adenosine triphosphate (ATP) with a final concentration of 16 μM and 3 μM of fluorescently labeled substrate No. 2 peptide (purchased from Jill Biochemical (Shanghai) Co., Ltd., Cat. No. 112394) to the above-mentioned 384-well reaction plate. The mixed solution starts with reaction. The reaction was performed at room temperature for 60 minutes, and 30 μL of stop detection solution (ethylenediaminetetraacetic acid was added to the buffer) was added to stop the kinase reaction. The conversion rate was read with Caliper's EZ Reader platform, and the inhibition rate was converted.

Inhibition rate %=(average % of conversion rate of positive control-average % of conversion rate of each compound)/(average % of conversion rate of positive control-average % of conversion rate of negative control).

Taking the log value of the concentration as the X axis and the inhibition rate % as the Y axis, the log(inhibitor) vs.response-Variable slope of the analysis software GraphPad Prism 5 was used to fit the dose-response curve to obtain the IC _{50 of each compound on the enzyme activity} values, see Table 1 for the results.

Table 1

Note: LOXO-292 is prepared with reference to the method of Example 163 of WO2018071447A1

BLU-667 was prepared with reference to the method of Example 130 of WO2017079140A1

The test results show that the compounds of the present invention have good inhibitory activity on RET kinase.

Test Example 2: Proliferation Inhibitory Activity on Stably Transduced Cells Ba/F3 KIF5B-RET, Ba/F3 KIF5B-RET-G810R

Used to determine the effect of compounds on the Ba/F3 KIF5B-RET cell line that stably expresses KIF5B-RET fusion protein in mouse primary B cells Ba/F3 in vitro and the Ba/F3 mouse primary B cell Ba/F3 stably expressing KIF5B-RET fusion protein (RET region). Proliferation inhibitory activity of Ba/F3 KIF5B-RET-G810R cell line with G810R point mutation).

Cell source: Ba/F3 KIF5B-RET cells were purchased from Kangyuan Bochuang Biotechnology (Beijing) Co., Ltd., item number KC-1041; Ba/F3 KIF5B-RET-G810R cells were purchased from Kangyuan Bochuang Biotechnology (Beijing) Co., Ltd. Company, Cat. No. KC-1448.

Taken at logarithmic growth phase of Ba / F3 KIF5B-RET, Ba / F3 KIF5B-RET cells were seeded in 96 well plates (135μl / well) and placed in a thermostatic 37 ℃ incubator containing 5% CO ₂ in 24 hours . The compound of the present invention, BLU-667 and LOXO-292 have been previously dissolved in dimethyl sulfoxide (DMSO) to prepare a 10 mM stock solution, and then diluted to the target concentration in another 96-well plate with complete medium when testing. 10 times, and then add 15 μl/well of each compound to the 96-well plate seeded with cells to reach the target concentration. Three replicate wells were set for each concentration, and a DMSO blank control was set. Continue to incubate for 72 hours at 37°C in a _{constant temperature incubator with 5% CO 2 .} Take out the inoculated 96-well cell culture plate from the carbon dioxide constant temperature incubator, add 20 μl of the thiazolyl blue (ie MTT) stock solution that has been dissolved in 0.9% physiological saline in advance to 5 mg/ml and filter sterilized into each well, and then inoculate at 37°C. Incubate for 4 hr in a constant temperature incubator containing 5% CO _{2 .} Add 100μl of triple solution to each well (10g of sodium dodecyl sulfate (SDS), 5ml of isobutanol, and 0.1ml of 36%-38% hydrochloric acid are dissolved in purified water to prepare a 100ml solution), incubate at 37°C until the precipitation is complete Dissolve, detect the optical density value OD at the wavelength of 570nm and carry out data processing to calculate the inhibition rate. The inhibition rate data were analyzed using GraphPad Prism 5.0 software, and the dose-response curve was fitted by nonlinear S-curve regression, and IC ₅₀ values were calculated therefrom. The results are shown in Table 2.

Inhibition rate% = [(OD _{72 hours the medium containing the DMSO control group compound} -OD _{72 hours)} / (OD _{72 hours the culture medium containing DMSO control} -OD _{0 hours the medium containing DMSO control group)]} × 100%.

Table 2

Note: ND means not determined

LOXO-292 was prepared with reference to the method of Example 163 of WO2018071447A1

BLU-667 was prepared with reference to the method of Example 130 of WO2017079140A1

The test results show that the compound of the present invention has good proliferation inhibitory activity on the stable transfection cells Ba/F3 KIF5B-RET and Ba/F3 KIF5B-RET-G810R; /F3 KIF5B-RET-G810R has better proliferation inhibitory activity.

Test Example 3: Growth Inhibitory Effect on Stably Transduced Cells Ba/F3 KIF5B-RET, Ba/F3 KIF5B-RET-G810R Nude Mouse Subcutaneously Transplanted Tumors

This experiment was used to study the growth inhibitory effect and safety of the compounds on subcutaneously transplanted tumors of Ba/F3 KIF5B-RET and Ba/F3 KIF5B-RET-G810R nude mice.

Cell culture: Cells stably transfected Ba / F3 KIF5B-RET, Ba / F3 KIF5B-RET-G810R was cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum, placed in a thermostatic containing 5% CO ₂ at 37 ℃ Cultivated in an incubator. Exponentially growing cells were harvested and counted for seeding.

Experimental animals: BALB/c Nude nude mice, female, 5 weeks old, 30, purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

For stably transfected cells Ba/F3 KIF5B-RET and stably transfected cells Ba/F3 KIF5B-RET-G810R, 3 experimental groups were set up, each with 5 mice; the 3 experimental groups were: containing 10% dimethylacetamide + Vehicle control group (ie Vehicle group), LOXO-292 group and Example 12 compound group of 5% polyethylene glycol-15 hydroxystearate+85% physiological saline.

Experimental plan 3A: Ba/F3 KIF5B-RET cell line stably transfected (2×10 ⁶ cells/cell) was inoculated subcutaneously on the right back of BALB/c Nude nude mice. tumor growth. When the tumors grew to be larger than 150 mm ³ , they were randomly divided into groups according to tumor size. The LOXO-292 group and the compound group of Example 12 were administered by intragastric administration at 10 mg/kg, and the administration volume was 10 uL/g. The vehicle control group was given the same amount of vehicle, twice a day, for 14 consecutive days. Throughout the experiment, the body weight and tumor size of the mice were measured twice a week to observe whether there was any toxicity.

Experimental protocol 3B: The stable transfected cells Ba/F3 KIF5B-RET-G810R cell line (2×10 ⁶ cells/cell) were inoculated subcutaneously on the right back of BALB/c Nude nude mice, and the inoculation volume for each mouse was 0.1 mL. Regular observation of tumor growth. When the tumors grew to be larger than 150 mm ³ , they were randomly divided into groups according to tumor size. The LOXO-292 group and the compound group of Example 12 were administered orally at 30 mg/kg, and the administration volume was 10 uL/g. The vehicle control group was given the same amount of vehicle, twice a day, for 14 consecutive days. Throughout the experiment, the body weight and tumor size of the mice were measured twice a week to observe whether there was any toxicity.

The formula for calculating tumor volume (TV) is: TV=1/2×a×b×b, where a and b represent the length and width of the tumor, respectively.

Results: Animals in the vehicle control group in experimental schemes 3A and 3B all died after 7 days, while no animals died in the LOXO-292 group and the compound group of Example 12, and there was no significant decrease in body weight during administration; in experimental schemes 3A and 3B, 3 The tumor volume change curves of the experimental groups are shown in Figure 1 and Figure 2, respectively.

The test results show that the compound of the present invention has a good inhibitory effect on the growth of stably transduced cells Ba/F3 KIF5B-RET and Ba/F3 KIF5B-RET-G810R nude mice subcutaneously transplanted tumor; Ba/F3 KIF5B-RET-G810R nude mice have a better inhibitory effect on the growth of subcutaneously transplanted tumors; the compounds of the present invention have little effect on the body weight of nude mice and show better safety.

All documents mentioned herein are incorporated by reference into this application. In addition, it should be noted that, after reading the above disclosure of the present application, those skilled in the art can make various modifications, changes or modifications to the present invention without departing from the spirit and scope of the present invention, but these variations should also be fall within the scope described in the appended claims of this application.

Claims (23)

1. A compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof,

   

   in:

   X is selected from CR ₆ or N;

   Y is selected from CR ₆ or N;

   Z is selected from CR ₆ or N;

   where 0, 1 or 2 of X, Y and Z are N;

   R _{1 is} selected from hydrogen, 6-10 membered aryl or 5-12 membered heteroaryl, wherein each of said 6-10 membered aryl or 5-12 membered heteroaryl is optionally replaced by 1, 2, 3 or 4 each independently selected from the group consisting of hydroxy, halogen, C _1-6 alkyl, C _1-6 alkoxy, halogenated C _1-6 alkyl, halogenated C _1-6 alkoxy, hydroxy C _1-6 alkoxy substituted with substituents of -NR ₇ R ₈ or -O-(CH ₂ ) _n -NR ₇ R ₈ ;

   R _{2 is} selected from hydrogen, C _1-4 alkyl or halogenated C _1-4 alkyl;

   R _{3 is} selected from hydrogen, C _1-6 alkyl, 3-6 membered cycloalkyl, 3-8 membered heterocyclyl or 5-12 membered heteroaryl, wherein said C _1-6 alkyl, 3-6 membered cycloalkyl, 3-8 membered heterocyclyl or 5-12 membered heteroaryl group each optionally substituted with two, three or four substituents each independently selected from halogen, C _1-4 alkyl, C _{1- 4} alkoxy, halogenated C _1-4 alkyl, halogenated C _1-4 alkoxy, cyano, hydroxy, hydroxy C _1-4 alkyl, -C(=O)-NR ₇ R ₈ , - SO ₂ -C _1-4 alkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclyl substituent, wherein the 5-6 membered heteroaryl or 3-6 membered heterocyclyl are each optionally by 1, 2 or 3 each independently selected from halogen, hydroxy, C _1-4 alkyl, C _1-4 alkoxy-(CH ₂ ) _m - or -C(=O)-C _{1 -4} alkyl substituents are substituted;

   R _{4 is} selected from hydrogen, C _1-4 alkyl or halogenated C _1-4 alkyl;

   R _{5 is} selected from hydrogen, halogen, cyano, C _1-4 alkyl or halogenated C _1-4 alkyl;

   each R _{6 is} independently selected from H, C _1-4 alkyl, C _1-4 alkoxy, haloC _1-4 alkyl or halogen;

   each R _{7 is} independently selected from H, C _1-4 alkyl or halogenated C _1-4 alkyl;

   each R _{8 is} independently selected from H, C _1-4 alkyl or halogenated C _1-4 alkyl;

   n is selected from 1, 2 or 3;

   m is selected from 0, 1, 2 or 3.
2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof as claimed in claim wherein, X is CR _6, Y is CR _6, Z is CR _6, wherein each R ₆ is H.
3. The compound according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof, wherein R _{1 is} selected from phenyl or 5-6 membered heteroaryl, wherein the phenyl or 5-6 membered heteroaryl aryl group each optionally substituted with 1, 2 or 3 substituents each independently selected from hydroxy, halo, C _1-6 alkyl, C _1-6 alkoxy, halo C _1-6 alkyl, halogeno C _{1 -6} Substituents of alkoxy or cyano.
4. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt thereof, wherein R _{1 is} selected from phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyrrolyl, imidazolyl or thiazolyl, wherein each of the phenyl, pyridyl, pyrimidinyl, pyrazinyl, pyrrolyl, imidazolyl or thiazolyl groups is optionally substituted by 1, 2 or 3 each independently selected from hydroxy, halogen, C _1-4 alkyl , C _1-4 alkoxy, halogenated C _1-4 alkyl or halogenated C _1-4 alkoxy substituents.
5. The compound according to any one of claims 1-4 or a pharmaceutically acceptable salt thereof, wherein R _{1 is} selected from phenyl,

   

   

   wherein the phenyl,

   

   

   Each optionally substituted with 1, 2 or 3 substituents each independently selected from hydroxy, F, Cl, Br, C 1-4 alkyl, C _1-4 alkoxy, halo C _1-4 alkyl or halo Substituents of C _{1-4 alkoxy.}
6. The compound according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof, wherein R _{1 is} selected from phenyl,

   

   

   wherein the phenyl,

   

   

   Each is optionally selected by 1, 2 or 3 each independently selected from hydroxy, F, Cl, Br, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, n-propoxy , isopropoxy, monofluoromethyl, difluoromethyl, trifluoromethyl, CF ₃ CH ₂ -, CHF ₂ CH ₂ -, CH ₂ FCH ₂ -, monofluoromethoxy, difluoromethoxy , trifluoromethoxy, CF ₃ CH ₂ -O-, CHF ₂ CH ₂ -O- or CH ₂ FCH ₂ -O- substituents.
7. The compound according to any one of claims 1-6, or a pharmaceutically acceptable salt thereof, wherein R _{1 is} selected from

   

   
8. The compound of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, wherein R _{2 is} selected from hydrogen or C _1-4 alkyl.
9. The compound of any one of claims 1-8, or a pharmaceutically acceptable salt thereof, wherein R _{4 is} selected from hydrogen or C _1-4 alkyl.
10. The compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, wherein R _{5 is} selected from hydrogen, halogen or cyano.
11. The compound according to any one of claims 1-10, or a pharmaceutically acceptable salt thereof, wherein R _{3 is} selected from hydrogen, C _1-6 alkyl or 3-6 membered cycloalkyl, wherein said C _{1 -6} alkyl or 3-6 membered cycloalkyl groups are each optionally selected by 1, 2 or 3 each independently selected from halogen, C _1-4 alkyl, C _1-4 alkoxy, halogenated C _{1- Substituted by substituents of 4} alkyl, halogenated C _1-4 alkoxy, cyano, hydroxyl, -SO ₂ -C _1-4 alkyl, 5-6-membered heteroaryl or 3-6-membered heterocyclic , wherein each of the 5-6 membered heteroaryl or 3-6 membered heterocyclyl is optionally substituted by 1, 2 or 3 substituents each independently selected from halogen, hydroxy or C _1-4 alkyl .
12. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein, R ₃ is selected from hydrogen, C _1-4 alkyl, cyclopropyl, cyclobutyl or cyclopentyl, wherein The C _1-4 alkyl, cyclopropyl, cyclobutyl or cyclopentyl groups are each optionally optionally substituted by 1, 2 or 3 each independently selected from F, Cl, Br, C _1-4 alkyl, C _1-4 alkoxy, halogenated C _1-4 alkyl, halogenated C _1-4 alkoxy, cyano, hydroxyl, -SO ₂ -C _1-4 alkyl,

    

    

    

    substituted by the substituents in which the

    

    

    

    Each is optionally substituted with 1, 2 or 3 substituents each independently selected from F, Cl, Br, hydroxy or _C1-4 alkyl.
13. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, wherein, R ₃ is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, , sec-butyl, tert-butyl, cyclopropyl, cyclobutyl, or cyclopentyl, wherein the methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec , tert-butyl, cyclopropyl, cyclobutyl, or cyclopentyl are each optionally substituted by 1, 2 or 3 each independently selected from F, Cl, Br, methyl, ethyl, methoxy, ethoxy group, monofluoromethyl, difluoromethyl, trifluoromethyl, CF ₃ CH ₂ -, CHF ₂ CH ₂ -, CH ₂ FCH ₂ -, monofluoromethoxy, difluoromethoxy, trifluoromethyl Oxy, CF ₃ CH ₂ -O-, CHF ₂ CH ₂ -O-, CH ₂ FCH ₂ -O-, cyano, hydroxyl, -SO ₂ -CH ₃ , -SO ₂ -CH ₂ CH ₃ ,

    

    

    

    substituted by the substituents in which the

    

    

    

    Each is optionally substituted with 1, 2 or 3 substituents each independently selected from F, Cl, Br, hydroxy, methyl or ethyl.
14. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 13, wherein, R ₃ is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, monofluoromethyl, difluoromethyl, trifluoromethyl,

    

    

    
15. The compound according to any one of claims 1-14 or a pharmaceutically acceptable salt thereof, which is a compound selected from the following structures:

    

    

    
16. The preparation method of compound shown in formula (I), it comprises:

    

    

    Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , X, Y, Z are as defined in any one of claims 1-14; L is a leaving group selected from halogen; P is an amino protecting group , each P can be the same or different, and each is independently selected from tert-butoxycarbonyl, benzyloxycarbonyl or benzyl;

    Using compound a-1 and compound a-2 as starting materials, Suzuki coupling reaction occurs to obtain compound a-3; compound a-3 undergoes substitution reaction with compound a-4 to obtain intermediate 1; intermediate 1 is deprotected to obtain compound a-5; compound a-5 undergoes reductive amination reaction with compound a-6 to obtain intermediate 2; intermediate 2 is combined with

    

    A coupling reaction occurs to obtain a compound represented by formula (I);

    Or intermediate 2 and P-NH ₂ undergo coupling reaction to obtain compound a-7; compound a-7 _{undergoes substitution reaction with R 3} -L to obtain compound a-8; compound a-8 is deprotected to obtain formula (I) compound.
17. The preparation method of compound shown in formula (I), it comprises:

    

    Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , X, Y, Z are as defined in any one of claims 1-14; L is a leaving group selected from halogen; P is an amino protecting group , each P can be the same or different, and each is independently selected from tert-butoxycarbonyl, benzyloxycarbonyl or benzyl;

    Intermediate 1 with

    

    A coupling reaction occurs to obtain compound b-1; compound b-1 is deprotected to obtain intermediate 3; intermediate 3 undergoes reductive amination reaction with compound a-6 to obtain a compound represented by formula (I);

    Or, compound b-2 is obtained by coupling reaction of intermediate 1 with P-NH _{2 ; compound b-2 is obtained by substitution reaction of compound b-2 with R 3} -L; compound b-3 is deprotected to obtain intermediate 3; Compound 3 is subjected to reductive amination reaction with compound a-6 to obtain the compound represented by formula (I).
18. an intermediate compound selected from compound a-7, compound a-8, compound b-1, compound b-2, compound b-3 or intermediate 3 shown in the following structure,

    

    

    Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , X, Y, Z are as defined in any one of claims 1-14; P is an amino protecting group, and each P may be the same or different, and each independently Selected from tert-butoxycarbonyl, benzyloxycarbonyl or benzyl.
19. A pharmaceutical composition comprising a compound of any one of claims 1-15, or a pharmaceutically acceptable salt thereof, and an optional pharmaceutically acceptable carrier.
20. Use of the compound of any one of claims 1-15 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 19 in the preparation of a medicament for the treatment of a disease mediated by RET.
21. The application according to claim 20, wherein the RET is selected from wild-type RET, mutant RET, and RET fusion; the mutant RET is selected from G810R mutant RET, M918T mutant RET, V804L mutant RET, V804M Mutant RET, the RET fusion is selected from KIF5B-RET fusion and CCDC6-RET fusion.
22. The use according to claim 20, wherein the disease is selected from cancer, irritable bowel syndrome.
23. Use of the compound of any one of claims 1-15 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 19 in the preparation of a medicament for treating cancer.

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