CN113461670B

CN113461670B - Novel compounds as inhibitors of rearrangement kinase during transfection

Info

Publication number: CN113461670B
Application number: CN202010242796.7A
Authority: CN
Inventors: 孔祥龙; 周超; 郑之祥
Original assignee: Nanjing Innocare Pharma Tech Co ltd
Current assignee: Nanjing Innocare Pharma Tech Co ltd
Priority date: 2020-03-31
Filing date: 2020-03-31
Publication date: 2024-08-09
Anticipated expiration: 2040-03-31
Also published as: WO2021197250A1; CN113461670A

Abstract

The present invention relates to compounds, pharmaceutical compositions containing them and methods for their preparation and their use as inhibitors of rearrangement (RET, REARRANGED DURING TRANSFECTION) kinase during transfection. The compound is a compound shown in a formula I or pharmaceutically acceptable salts, prodrugs, isomers and stable isotope derivatives thereof. The invention also relates to the use of said compounds for the treatment or prophylaxis of diseases which are associated with RET kinase, such as tumors, and to methods of using the same for the treatment of said diseases.

Description

Novel compounds as inhibitors of rearrangement kinase during transfection

Technical Field

The present invention relates to compounds, pharmaceutical compositions containing them and their use as inhibitors of rearrangement (RET, REARRANGED DURING TRANSFECTION) kinase during transfection. More particularly, the present invention provides novel compounds that are inhibitors of RET kinase, pharmaceutical compositions containing such compounds, and methods of using the compounds to treat or prevent diseases associated with RET kinase, such as tumors. The invention also relates to a process for preparing the compounds described below.

Background

The RET (Rearranged during transfection) gene encodes a membrane receptor tyrosine kinase RET protein, belongs to the cadherin superfamily, is expressed in cells of neural crest origin and genitourinary system, and plays a vital role in neural crest development. RET kinase activates the Ras/MAPK, PI3K/Akt, JNK, p38 and PLCg pathway signaling cascades by binding to one of four glial cell-derived neurotrophic factor (GDNF) family receptor alpha (gfrα) proteins to form a complex that homodimerizes, phosphorylates and activates its tyrosine kinase activity (Mulligan, l.m. Nature REVIEWS CANCER, 2014, 14, 173-186).

Oncogenic, activating mutations (mainly rearrangements or fusions at the cytogenetic level) of the RET gene amplify the signaling cascade without relying on ligand binding, while also activating other signaling cascades (such as STAT3 and STAT 1) responses, promoting tumorigenesis. RET is a cancer driver, and mutations result in overactivity of the RET signaling pathway, leading to uncontrolled cellular growth, which in turn triggers tumor formation.

RET protein mutation types mainly comprise fusion mutation with KIF5B, TRIM, CCDC6, NCOA4 and other genes, point mutation of M918T and other sites, common RET mutation mainly occurs in thyroid cancer, non-small cell lung cancer and other cancer types, although the incidence rate of RET mutation in non-small cell lung cancer is only 2%, the patient base of China is very large, the patients are not rare in clinical treatment, RET fusion is more common in young patients, particularly young non-smoking lung adenocarcinoma patients, and the incidence rate is up to 7% -17%.

In the prior art of treating RET positive patients, the targeted drug often selects cabatinib or vandetanib, and the two drugs are used as multi-target tyrosine kinase inhibitors, so that the selectivity of RET is not strong. In 26 small samples of cabatinib with RET treatment, the primary endpoint ORR was 28% and the median time of PFS was 5.5 months, while 19 patients in treatment had adjusted dose due to adverse effects and the toxic response was very pronounced. Development of selective RET kinase inhibitors has the potential to reduce adverse reactions caused by off-target sites and improve therapeutic effects. Currently, there is a need for new selective RET kinase inhibitors that reduce the adverse effects caused by off-target sites and improve the therapeutic effect, mainly by developing selective RET kinase inhibitors of Blu-667 (US 20170121312A1, subbiah, v.; gainor, j.f.; et al Cancer discovery.2018, 8 (7), 836-849).

Disclosure of Invention

The object of the present invention is to provide a compound of formula (I), an isomer, a prodrug, a stable isotope derivative or a pharmaceutically acceptable salt thereof, which is useful as a selective RET kinase inhibitor:

Wherein:

x ¹ is CR ¹ or N; preferably CH or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

y ¹ is CR ² or N; preferably CH or N; most preferably N;

Y ² is CR ³ or N; preferably CH or N; most preferably CH;

Y ³ is CR ⁴ or N; preferably CH or N;

Y ⁴ is CR ⁵ or N; preferably CH or N; most preferably CH;

Provided that up to 2 of Y ¹、Y²、Y³ and Y ⁴ are N;

Preferably one of the following conditions: y ¹-Y⁴ is not N; only Y ¹ or Y ² is N; y ¹ and Y ² are both N; or Y ¹ and Y ³ are both N;

Further preferred is one of the following conditions: only Y ¹ or Y ² is N; or Y ¹ and Y ³ are both N; most preferably one of the following conditions: y ¹ is N, Y ²、Y³ and Y ⁴ are CH; or Y ¹ and Y ³

N, Y ² and Y ⁴ are CH;

R ¹-R⁵ is each independently selected from hydrogen, halogen, cyano, C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl, heteroaryl 、-OR⁶、-NR⁷R⁸、-OC(O)NR⁷R⁸、-C(O)OR⁶、-C(O)NR⁷R⁸、-NR⁹C(O)R⁶、-NR⁹C(O)NR⁷R⁸、-S(O)_mR⁶、-NR⁹S(O)_mR⁶、-SR⁶、-S(O)_mNR⁷R⁸, or-NR ⁹S(O)_mNR⁷R⁸, wherein the alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkenyl, or alkynyl is optionally substituted with one or more substituents selected from halogen, cyano, C3-C8 cycloalkyl, 4-8 membered heterocyclyl 、-OR¹⁰、-NR¹¹R¹², -OC(O)NR¹¹R¹²、-C(O)OR¹⁰、-C(O)NR¹¹R¹²、-NR¹¹C(O)R¹⁰、-NR¹³C(O)NR¹¹R¹²、-S(O)_mR¹⁰、-NR¹³S(O)_mR¹⁰、-SR¹⁰、-S(O)_mNR¹¹R¹², or-NR ¹³S(O)_mNR¹¹R¹²; preferably, R ¹-R⁵ is each independently selected from hydrogen, halogen, cyano, C1-C6 alkyl, -OR ⁶, OR-NR ⁷R⁸; further preferably, each R ¹-R⁵ is independently selected from hydrogen, halogen, or C1-C4 alkyl; most preferably, R ¹-R⁵ are both hydrogen;

A is selected from hydrogen, halogen, cyano or C1-C8 alkyl; preferably, A is selected from hydrogen, halogen, cyano or C1-C6 alkyl; further preferably, A is selected from hydrogen, cyano or C1-C4 alkyl; most preferably, a is hydrogen;

b is selected from hydrogen, halogen, cyano, C1-C4 alkyl, -OR ¹⁴, OR optionally substituted Ar ¹; preferably, B is selected from hydrogen, halogen, -OR ¹⁴, OR optionally substituted Ar ¹; most preferably, B is selected from hydrogen, halogen, -OR ¹⁴, OR ；

R ¹⁴ is selected from hydrogen, C1-C8 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl, heteroaryl, alkenyl, or alkynyl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkenyl, alkynyl is optionally substituted with one or more substituents selected from halogen, cyano, hydroxy, C3-C8 cycloalkyl, 4-8 membered heterocyclyl optionally substituted with C1-C6 alkyl; preferably, R ¹⁴ is selected from hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, or 4-8 membered heterocyclyl, wherein said alkyl, cycloalkyl, heterocyclyl is optionally substituted with one or more substituents selected from halogen, hydroxy, C3-C8 cycloalkyl, 4-8 membered heterocyclyl optionally substituted with C1-C6 alkyl; further preferably, R ¹⁴ is selected from C1-C4 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclyl, wherein said alkyl, cycloalkyl, heterocyclyl is optionally substituted with one or more substituents selected from halogen, hydroxy, 4-6 membered heterocyclyl optionally substituted with C1-C4 alkyl; most preferably, R ¹⁴ is C1-C4 alkyl optionally substituted with 1-methylpiperidin-4-yl;

Ar ¹ is selected from 5-or 6-membered heteroaryl groups containing 1-3 ring heteroatoms, wherein each heteroatom is independently selected from N, O, S; preferably, ar ¹ is selected from 5-or 6-membered heteroaryl groups containing two ring N atoms; further preferably, ar ¹ is selected from the group consisting of 5-membered heteroaryl groups containing two ring N atoms; most preferably Ar ¹ is ；

Ar ¹ may be optionally substituted with one or more substituents each independently selected from halogen, cyano, C1-C8 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl, heteroaryl, C2-C8 alkenyl, C2-C8 alkynyl 、-OR⁶、-NR⁷R⁸、-OC(O)NR⁷R⁸、-C(O)OR⁶、-C(O)R⁶、-C(O)NR⁷R⁸、-NR⁹C(O)R⁶、-NR⁹C(O)NR⁷R⁸、-S(O)_mR⁶、-NR⁹S(O)_mR⁶、-SR⁶、-S(O)_mNR⁷R⁸ or-NR ⁹S(O)_mNR⁷R⁸, wherein the alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkenyl, alkynyl is optionally substituted with one or more substituents selected from halogen, cyano, C3-C8 cycloalkyl, 4-8 membered heterocyclyl 、-OR¹⁰、-NR¹¹R¹², -OC(O)NR¹¹R¹²、-C(O)OR¹⁰、-C(O)NR¹¹R¹²、-NR¹¹C(O)R¹⁰、-NR¹³C(O)NR¹¹R¹²、-S(O)_mR¹⁰、-NR¹³S(O)_mR¹⁰、-SR¹⁰、-S(O)_mNR¹¹R¹²、-NR¹³S(O)_mNR¹¹R¹²;

R ¹⁵ is selected from hydrogen, C1-C8 alkyl, hydroxy C1-C8 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl or heteroaryl; preferably, R ¹⁵ is selected from hydrogen, C1-C6 alkyl, hydroxyC 1-C6 alkyl, C3-C8 cycloalkyl or 4-8 membered heterocyclyl; further preferred, R ¹⁵ is selected from hydrogen, C1-C4 alkyl, hydroxyC 1-C4 alkyl, C3-C6 cycloalkyl or 4-6 membered heterocyclyl containing 1-2 heteroatoms selected from N, O or S; most preferably, R ¹⁵ is C1-C4 alkyl;

D is selected from 4-8 membered heterocyclylene, 6-8 membered fused heterocyclylene or 7-11 membered spiroheterocyclylene containing 1-3 heteroatoms selected from N, O; preferably, D is selected from 4-6 membered heterocyclylene containing 1-2N or 6-8 membered fused heterocyclylene containing 1-2N; most preferably, D is selected from:

; and when X ² is CH, D is not Or (b)；

D is optionally substituted with a substituent selected from halogen, cyano, hydroxy, amino or C1-C8 alkyl, wherein said alkyl is optionally substituted with a substituent selected from the group consisting of: halogen, hydroxy, mono-or di (C1-C8 alkyl) amino, N- (C1-C8 alkyl) -N- (C1-C4 alkylcarbonyl) amino, 4-8 membered heterocyclyl optionally substituted by halogen, hydroxy, (C1-C4 alkyl) carbonyl or C1-C8 alkyl; preferably, D is optionally substituted with one substituent selected from halogen, cyano, hydroxy, amino or C1-C6 alkyl, wherein said alkyl is optionally substituted with one substituent selected from the group consisting of: halogen, hydroxy, mono-or di (C1-C6 alkyl) amino, N- (C1-C6 alkyl) -N- (C1-C4 alkylcarbonyl) amino, 4-6 membered heterocyclyl optionally substituted by halogen, hydroxy, (C1-C4 alkyl) carbonyl or C1-C6 alkyl; further preferred, D is optionally substituted with one substituent selected from hydroxy, amino or C1-C6 alkyl, wherein said alkyl is optionally substituted with one substituent selected from the group consisting of: hydroxy, mono-or di (C1-C4 alkyl) amino, N- (C1-C4 alkyl) -N- (C1-C4 alkylcarbonyl) amino, 4-6 membered heterocyclyl optionally substituted by (C1-C4 alkyl) carbonyl or C1-C6 alkyl; most preferably, D is optionally substituted with one substituent selected from hydroxy, amino or C1-C4 alkyl, said alkyl being optionally substituted with one substituent selected from hydroxy, di (C1-C4 alkyl) amino, N-acetyl-N-methylamino, morpholin-4-yl, 1-ethylpiperazin-4-yl or 1-acetylpiperazin-4-yl; wherein for the followingIf the substituent is hydroxy, it is substituted at its 3-position; if the substituent is other than hydroxy, then it is substituted at the 4-position; for the followingThe substituent is hydroxyl or amino, and is substituted at the 4-position;

E is selected from amino, NHC (O) R ^X、-C(O)R^y, 、-OR⁶、-NR⁷R⁸、-OC(O)NR⁷R⁸、-C(O)OR⁶、-C(O)NR⁷R⁸、-NR⁹C(O)NR⁷R⁸、-S(O)_mR⁶、-NR⁹S(O)_mR⁶、-SR⁶、-S(O)_mNR⁷R⁸、-NR⁹S(O)_mNR⁷R⁸、 Or-CH ₂-Ar²; preferably E is selected from amino, heteroaryloxy, -NHC (O) R ^X、-SO₂ (C1-C8) alkyl, -C (O) R ^y,Or-CH ₂-Ar²; further preferably E is selected from amino, pyridyloxy, -NHC (O) R ^X、SO₂ (C1-C6) alkyl, -C (O) R ^y,Or-CH ₂-Ar²;

most preferably, when D is When E is selected from amino, -NHC (O) R ^X or-SO ₂ (C1-C4) alkyl; when D isWhen E is pyridine-2-oxy; when D is When E is selected from the group consisting of-C (O) R ^y,-SO ₂ (C1-C4) alkyl, or-CH ₂-Ar²;

r ^X is selected from C1-C8 alkoxy, optionally substituted aryl, optionally substituted heteroaryl containing 1 or 2 heteroatoms selected from N, O, S, optionally substituted C1-C8 alkyl, optionally substituted 4-8 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, S, C3-C8 cycloalkyl or amino optionally substituted by C1-C8 alkyl; the optional substituent is selected from halogen, nitro, cyano, hydroxy, C1-C8 alkoxy or C3-C8 cycloalkyl; preferably, R ^X is selected from C1-C6 alkoxy, optionally substituted 6 membered aryl, optionally substituted 5-6 membered heteroaryl containing 1 or 2 heteroatoms selected from N, O, optionally substituted C1-C6 alkyl, optionally substituted 4-6 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, S, C3-C6 cycloalkyl or amino optionally substituted by C1-C6 alkyl; the optional substituents are selected from halogen, hydroxy, C1-C6 alkoxy or C3-C6 cycloalkyl; most preferably, R ^X is selected from C1-C4 alkoxy, optionally mono-or di-substituted phenyl, optionally mono-or di-substituted pyridinyl, 5 membered heteroaryl containing 1 or 2 heteroatoms selected from N, O, optionally substituted C1-C4 alkyl, optionally substituted 4-6 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, C3-C6 cycloalkyl or amino optionally substituted by C1-C4 alkyl; the optional substituents are selected from halogen, hydroxy, C1-C4 alkoxy or C3-C6 cycloalkyl;

R ^y is selected from optionally substituted amino, C1-C8 alkyl, 4-8 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, S, or C3-C8 cycloalkyl, said substituents being selected from C1-C8 alkyl optionally substituted with aryl or heteroaryl; preferably, R ^y is selected from optionally substituted amino, C1-C6 alkyl, 4-6 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, S, or C3-C6 cycloalkyl, said substituents being selected from C1-C6 alkyl optionally substituted by aryl; most preferably, R ^y is selected from amino optionally substituted by C1-C4 alkyl or benzyl, C1-C4 alkyl, 4-6 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, or C3-C6 cycloalkyl;

Ar ² is selected from optionally substituted aryl or heteroaryl, said substituents being selected from halogen, hydroxy, amino or C1-C8 alkoxy; preferably Ar ² is selected from 5-6 membered aryl or heteroaryl optionally mono-or di-substituted, said substituents being selected from halogen, C1-C6 alkoxy; most preferably Ar ² is selected from phenyl optionally mono-or disubstituted, pyridinyl optionally mono-or disubstituted, said substituents being selected from halogen, C1-C4 alkoxy;

R ^m、Rⁿ is each independently selected from hydrogen, hydroxy, C1-C8 alkyl or hydroxy C1-C8 alkyl, or R ^m、Rⁿ together with the carbon atom to which it is attached form C3-C8 cycloalkyl; preferably, R ^m、Rⁿ is each independently selected from hydrogen, hydroxy, C1-C6 alkyl or hydroxyC 1-C6 alkyl, or R ^m、Rⁿ together with the carbon atom to which it is attached form C3-C6 cycloalkyl; most preferably, each R ^m、Rⁿ is independently selected from hydrogen, hydroxy or hydroxy C1-C4 alkyl, or R ^m、Rⁿ together with the carbon atom to which it is attached form cyclopropyl;

R ⁶、R⁷、R⁸、R⁹、R¹⁰、R¹¹、R¹²、R¹³ is each independently selected from hydrogen, halogen, hydroxy, amino, C1-C8 alkyl, C1-C8 alkoxy, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, heteroaryl, aryl, C2-C8 alkenyl, or C2-C8 alkynyl;

r is selected from 0, 1, 2 or 3; preferably r is selected from 0, 1 or 2;

m is selected from 1 or 2.

Preferably, the present invention provides a compound of formula (I), an isomer, a prodrug, a stable isotope derivative or a pharmaceutically acceptable salt thereof, as hereinbefore described, useful as a selective RET kinase inhibitor, wherein

X ¹ is CR ¹ or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

Y ¹ is CR ² or N;

y ² is CR ³ or N;

y ³ is CR ⁴ or N;

Y ⁴ is CR ⁵ or N;

And one of the following conditions is satisfied: y ¹-Y⁴ is not N; only Y ¹ or Y ² is N; y ¹ and Y ² are both N; or Y ¹ and Y ³ are both N;

r ¹-R⁵ is each independently selected from hydrogen, halogen, cyano, C1-C8 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl, heteroaryl, C2-C8 alkenyl, C2-C8 alkynyl 、-OR⁶、-NR⁷R⁸、-OC(O)NR⁷R⁸、-C(O)OR⁶、-C(O)NR⁷R⁸、

-NR⁹C(O)R⁶、-NR⁹C(O)NR⁷R⁸、-S(O)_mR⁶、-NR⁹S(O)_mR⁶、-SR⁶、-S(O)_mNR⁷R⁸ Or-NR ⁹S(O)_mNR⁷R⁸;

a is selected from hydrogen, halogen, cyano or C1-C6 alkyl;

B is selected from hydrogen, halogen, -OR ¹⁴, OR optionally substituted Ar ¹;

R ¹⁴ is selected from hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl, heteroaryl, alkenyl, or alkynyl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkenyl, alkynyl is optionally substituted with one or more substituents selected from the group consisting of: halogen, cyano, hydroxy, C3-C8 cycloalkyl, 4-8 membered heterocyclyl optionally substituted by C1-C6 alkyl;

ar ¹ is selected from 5-or 6-membered heteroaryl groups containing 1-3 ring heteroatoms, wherein each heteroatom is independently selected from N, O, S;

Ar ¹ is optionally substituted with one or more substituents each independently selected from halogen, cyano, C1-C8 alkyl, C3-C8 cycloalkyl, heterocyclyl, aryl, heteroaryl, C2-C8 alkenyl, C2-C8 alkynyl 、-OR⁶、-NR⁷R⁸、-OC(O)NR⁷R⁸、-C(O)OR⁶、-C(O)R⁶、-C(O)NR⁷R⁸、-NR⁹C(O)R⁶、-NR⁹C(O)NR⁷R⁸、-S(O)_mR⁶、-NR⁹S(O)_mR⁶、-SR⁶、-S(O)_mNR⁷R⁸ or-NR ⁹S(O)_mNR⁷R⁸;

D is selected from a 4-6 membered heterocyclylene group containing 1-2N or a 6-8 membered fused heterocyclylene group containing 1-2N, and when X ² is CH, D is not Or (b)；

D is optionally substituted with a substituent selected from halogen, cyano, hydroxy, amino, or C1-C8 alkyl, wherein the alkyl is optionally substituted with a substituent selected from: halogen, hydroxy, mono-or di (C1-C8 alkyl) amino, N- (C1-C8 alkyl) -N- (C1-C4 alkylcarbonyl) amino, 4-8 membered heterocyclyl optionally substituted by halogen, hydroxy, (C1-C4 alkyl) carbonyl or C1-C8 alkyl;

E is selected from amino, NHC (O) R ^X、-C(O)R^y, 、-OR⁶、-NR⁷R⁸、-OC(O)NR⁷R⁸、-C(O)OR⁶、-C(O)NR⁷R⁸、-NR⁹C(O)NR⁷R⁸、-S(O)_mR⁶、-NR⁹S(O)_mR⁶、-SR⁶、-S(O)_mNR⁷R⁸、-NR⁹S(O)_mNR⁷R⁸、 Or-CH ₂-Ar²;

r ^X is selected from C1-C8 alkoxy, optionally substituted aryl, optionally substituted heteroaryl containing 1 or 2 heteroatoms selected from N, O, S, optionally substituted C1-C8 alkyl, optionally substituted 4-8 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, S, C3-C8 cycloalkyl or amino optionally substituted by C1-C8 alkyl; the optional substituent is selected from halogen, nitro, cyano, hydroxy, C1-C8 alkoxy or C3-C8 cycloalkyl;

r ^y is selected from optionally substituted amino, C1-C8 alkyl, 4-8 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, S, or C3-C8 cycloalkyl, said substituents being selected from C1-C8 alkyl optionally substituted with aryl or heteroaryl;

Ar ² is selected from optionally substituted aryl or heteroaryl, said substituents being selected from halogen, hydroxy, amino or C1-C8 alkoxy;

R ^m、Rⁿ is each independently selected from hydrogen, hydroxy, C1-C8 alkyl, hydroxy C1-C8 alkyl, or R ^m、Rⁿ taken together with the carbon atom to which it is attached form C3-C8 cycloalkyl;

R ⁶、R⁷、R⁸、R⁹ is each independently selected from hydrogen, halogen, hydroxy, amino, C1-C8 alkyl, C1-C8 alkoxy, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, heteroaryl, aryl, C2-C8 alkenyl, or C2-C8 alkynyl;

m is 1 or 2;

r is 0, 1,2 or 3.

Further preferably, the present invention provides a compound of formula (I), an isomer, a prodrug, a stable isotope derivative or a pharmaceutically acceptable salt thereof, which is useful as a selective RET kinase inhibitor as described above, wherein

X ¹ is CR ¹ or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

Y ¹ is CR ² or N;

y ² is CR ³ or N;

y ³ is CR ⁴ or N;

Y ⁴ is CR ⁵ or N;

R ¹-R⁵ is each independently selected from hydrogen, halogen, cyano, C1-C6 alkyl, -OR ⁶, OR-NR ⁷R⁸;

a is selected from hydrogen, halogen, cyano or C1-C6 alkyl;

R ¹⁴ is selected from hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl, heteroaryl, alkenyl, or alkynyl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkenyl, alkynyl is optionally substituted with one or more substituents selected from the group consisting of: halogen, cyano, hydroxy, C3-C8 cycloalkyl, or 4-8 membered heterocyclyl optionally substituted with C1-C6 alkyl;

Ar ¹ is selected from 5-or 6-membered heteroaryl groups containing two ring N atoms;

Ar ¹ is optionally substituted with one or more substituents each independently selected from halogen, cyano, C1-C8 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl, heteroaryl, C2-C8 alkenyl, C2-C8 alkynyl 、-OR⁶、-NR⁷R⁸、-OC(O)NR⁷R⁸、-C(O)OR⁶、-C(O)R⁶、-C(O)NR⁷R⁸、-NR⁹C(O)R⁶、-NR⁹C(O)NR⁷R⁸、-S(O)_mR⁶、-NR⁹S(O)_mR⁶、-SR⁶、-S(O)_mNR⁷R⁸ or-NR ⁹S(O)_mNR⁷R⁸;

D is optionally substituted with a substituent selected from halogen, cyano, hydroxy, amino, C1-C8 alkyl, wherein said alkyl is optionally substituted with a substituent selected from the group consisting of: halogen, hydroxy, mono-or di (C1-C8 alkyl) amino, N- (C1-C8 alkyl) -N- (C1-C4 alkylcarbonyl) amino, 4-8 membered heterocyclyl optionally substituted by halogen, hydroxy, (C1-C4 alkyl) carbonyl or C1-C8 alkyl;

E is selected from amino, heteroaryloxy, -NHC (O) R ^X、-SO₂ (C1-C8) alkyl, -C (O) R ^y, Or-CH ₂-Ar²;

Ar ² is selected from optionally substituted aryl or heteroaryl, said substituents being selected from halogen, hydroxy, amino, C1-C8 alkoxy;

m is 1 or 2;

r is 0, 1,2 or 3.

Still further preferably, the present invention provides a compound of formula (I), an isomer, a prodrug, a stable isotope derivative or a pharmaceutically acceptable salt thereof, which is useful as a selective RET kinase inhibitor as described above, wherein

X ¹ is CR ¹ or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

Y ¹ is CR ² or N;

y ² is CR ³ or N;

y ³ is CR ⁴ or N;

Y ⁴ is CR ⁵ or N;

and one of the following conditions is satisfied: y ¹-Y⁴ is not N; only Y ¹ or Y ² is N; or Y ¹ and Y ³ are both N;

R ¹-R⁵ is each independently selected from hydrogen, halogen or C1-C4 alkyl;

a is selected from hydrogen, cyano or C1-C4 alkyl;

r ¹⁴ is selected from hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl, heteroaryl, alkenyl, or alkynyl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, alkenyl, alkynyl is optionally substituted with one or more substituents selected from the group consisting of: halogen, cyano, hydroxy, C3-C8 cycloalkyl, 4-8 membered heterocyclyl optionally substituted by C1-C6 alkyl;

ar ¹ is selected from the group consisting of 5-membered heteroaryl groups containing two ring N atoms;

Ar ¹ is optionally substituted with one or more substituents each independently selected from halogen, cyano, C1-C8 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl, heteroaryl, C2-C8 alkenyl, C2-C8 alkynyl 、-OR⁶、-NR⁷R⁸、-OC(O)NR⁷R⁸、-C(O)OR⁶、-C(O)R⁶、-C(O)NR⁷R⁸、-NR⁹C(O)R⁶、-NR⁹C(O)NR⁷R⁸、-S(O)_mR⁶、-NR⁹S(O)_mR⁶、-SR⁶、-S(O)_mNR⁷R⁸、-NR⁹S(O)_mNR⁷R⁸;

D is selected from a 4-6 membered heterocyclylene group containing 1-2N or a 6-8 membered fused heterocyclylene group containing 1-2N, and when X ² is CH, D is notOr (b)；

D is optionally substituted with a substituent selected from halogen, cyano, hydroxy, amino, C1-C6 alkyl, wherein said alkyl is optionally substituted with a substituent selected from the group consisting of: halogen, hydroxy, mono-or di (C1-C6 alkyl) amino, N- (C1-C6 alkyl) -N- (C1-C4 alkylcarbonyl) amino, 4-6 membered heterocyclyl optionally substituted by halogen, hydroxy, (C1-C4 alkyl) carbonyl or C1-C6 alkyl;

m is 1 or 2;

r is 0, 1,2 or 3.

X ¹ is CH or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

y ¹ is CH or N;

Y ² is CH or N;

y ³ is CH or N;

Y ⁴ is CH or N;

And one of the following conditions is satisfied: y ¹、Y²、Y³ and Y ⁴ are both CH; only Y ¹ or Y ² is N; or Y ¹ and Y ³ are both N;

a is hydrogen;

b is selected from hydrogen, halogen, -OR ¹⁴ OR ；

R ¹⁴ is selected from hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl, aryl, heteroaryl, alkenyl, or alkynyl, wherein said alkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkenyl, or alkynyl is optionally substituted with one or more substituents selected from the group consisting of: halogen, cyano, hydroxy, C3-C8 cycloalkyl, 4-8 membered heterocyclyl optionally substituted by C1-C6 alkyl;

r ¹⁵ is selected from hydrogen, C1-C8 alkyl, hydroxyC 1-C8 alkyl, C3-C8 cycloalkyl, 4-8 membered heterocyclyl containing 1-2 heteroatoms selected from N, O, S, aryl or heteroaryl;

d is selected from:

And when X ² is CH, D is not Or (b)；

D is optionally substituted with a substituent selected from halogen, cyano, hydroxy, amino or C1-C6 alkyl, wherein said alkyl is optionally substituted with a substituent selected from the group consisting of: halogen, hydroxy, mono-or di (C1-C6 alkyl) amino, N- (C1-C6 alkyl) -N- (C1-C4 alkylcarbonyl) amino, 4-6 membered heterocyclyl optionally substituted by halogen, hydroxy, (C1-C4 alkyl) carbonyl or C1-C6 alkyl;

E is selected from amino, pyridyloxy, -NHC (O) R ^X、SO₂ (C1-C6) alkyl, -C (O) R ^y, Or-CH ₂-Ar²;

r is 0, 1,2 or 3.

X ¹ is CH or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

y ¹ is CH or N;

Y ² is CH or N;

y ³ is CH or N;

Y ⁴ is CH or N;

a is hydrogen;

b is selected from hydrogen, halogen, -OR ¹⁴ OR ；

R ¹⁴ is selected from hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, or 4-8 membered heterocyclyl, wherein said alkyl, cycloalkyl, heterocyclyl is optionally substituted with one or more substituents selected from the group consisting of: halogen, hydroxy, C3-C8 cycloalkyl, 4-8 membered heterocyclyl optionally substituted by C1-C6 alkyl;

R ¹⁵ is selected from hydrogen, C1-C6 alkyl, hydroxyC 1-C6 alkyl, C3-C8 cycloalkyl, or 4-8 membered heterocyclyl containing 1-2 heteroatoms selected from N, O, S;

d is selected from:

And when X ² is CH, D is not Or (b)；

D is optionally substituted with a substituent selected from hydroxy, amino or C1-C6 alkyl, wherein said alkyl is optionally substituted with a substituent selected from: mono-or di (C1-C6 alkyl) amino, N- (C1-C6 alkyl) -N- (C1-C4 alkyl) carbonylamino, 4-6 membered heterocyclyl optionally substituted by C1-C6 alkyl, acetyl;

r ^X is selected from C1-C6 alkoxy, optionally substituted 6 membered aryl, optionally substituted 5-6 membered heteroaryl containing 1 or 2 heteroatoms selected from N, O, optionally substituted C1-C6 alkyl, optionally substituted 4-6 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, S, C3-C6 cycloalkyl or amino optionally substituted by C1-C6 alkyl; the optional substituents are selected from halogen, hydroxy, C1-C6 alkoxy or C3-C6 cycloalkyl;

R ^y is selected from optionally substituted amino, C1-C6 alkyl, 4-6 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, S, or C3-C6 cycloalkyl, said substituents being selected from C1-C6 alkyl optionally substituted by aryl;

Ar ² is selected from 5-6 membered aryl or heteroaryl optionally mono-or di-substituted, said substituents being selected from halogen or C1-C6 alkoxy;

R ^m、Rⁿ is each independently selected from hydrogen, hydroxy, C1-C6 alkyl or hydroxy C1-C6 alkyl, or R ^m、Rⁿ together with the carbon atom to which it is attached form C3-C6 cycloalkyl;

r is 0, 1,2 or 3.

X ¹ is CH or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

Y ¹ is N, Y ²、Y³ and Y ⁴ are CH; or Y ¹ and Y ³ are N, Y ² and Y ⁴ are CH;

a is hydrogen;

b is selected from hydrogen, halogen, -OR ¹⁴ OR ；

R ¹⁴ is selected from C1-C4 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclyl, wherein said alkyl, cycloalkyl, or heterocyclyl is optionally substituted with one or more substituents selected from the group consisting of: halogen, hydroxy, 4-6 membered heterocyclyl optionally substituted by C1-C4 alkyl;

R ¹⁵ is selected from hydrogen, C1-C4 alkyl, hydroxyC 1-C4 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclyl containing 1-2 heteroatoms each independently selected from N, O or S;

d is selected from:

And when X ² is CH, D is not Or (b)；

D is optionally substituted with a substituent selected from hydroxy, amino or C1-C4 alkyl, said alkyl being optionally substituted with a substituent selected from hydroxy, di (C1-C4 alkyl) amino, N-acetyl-N-methylamino, morpholin-4-yl, 1-ethylpiperazin-4-yl or 1-acetylpiperazin-4-yl;

R ^X is selected from C1-C4 alkoxy, optionally mono-or di-substituted phenyl, optionally mono-or di-substituted pyridinyl, 5 membered heteroaryl containing 1 or 2 heteroatoms selected from N, O, optionally substituted C1-C4 alkyl, optionally substituted 4-6 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, C3-C6 cycloalkyl or amino optionally substituted by C1-C4 alkyl; the optional substituents are selected from halogen, hydroxy, C1-C4 alkoxy or C3-C6 cycloalkyl;

R ^y is selected from amino optionally substituted by C1-C4 alkyl or benzyl, C1-C4 alkyl, 4-6 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, or C3-C6 cycloalkyl;

Ar ² is selected from phenyl optionally mono-or disubstituted, pyridinyl optionally mono-or disubstituted, said substituents being selected from halogen or C1-C4 alkoxy;

R ^m、Rⁿ is each independently selected from hydrogen, hydroxy C1-C4 alkyl, or R ^m、Rⁿ taken together with the carbon atom to which it is attached forms cyclopropyl;

r is 0, 1 or 2.

X ¹ is CH or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

a is hydrogen;

b is selected from hydrogen, halogen, -OR ¹⁴ OR ；

R ¹⁴ is selected from C1-C4 alkyl optionally substituted by 1-methylpiperidin-4-yl;

R ¹⁵ is C1-C4 alkyl;

D is selected from

And when X ² is CH, D is notOr (b)；

D is optionally substituted with one substituent selected from hydroxy, amino or C1-C4 alkyl, said alkyl being optionally substituted with one substituent selected from hydroxy, di (C1-C4 alkyl) amino, N-acetyl-N-methylamino, morpholin-4-yl, 1-ethylpiperazin-4-yl or 1-acetylpiperazin-4-yl;

wherein for the following If the substituent is hydroxy, it is substituted at its 3-position; if the substituent is other than hydroxy, then it is substituted at the 4-position; for the followingThe substituent is hydroxyl or amino, and is substituted at the 4-position;

When D is When E is selected from amino, -NHC (O) R ^X or-SO ₂ (C1-C4) alkyl;

When D is When E is pyridine-2-oxy;

When D is Or (b)When E is selected from the group consisting of-C (O) R ^y,-SO ₂ (C1-C4) alkyl, or-CH ₂-Ar²;

Ar ² is selected from phenyl optionally mono-or disubstituted, pyridinyl optionally mono-or disubstituted, said substituents being selected from halogen, C1-C4 alkoxy;

r is 0, 1 or 2.

Still further preferably, the present invention provides a compound of formula (I), an isomer, a prodrug, a stable isotope derivative or a pharmaceutically acceptable salt thereof, as described hereinbefore, useful as a selective RET kinase inhibitor, wherein the compound is:

。

The invention further relates to a pharmaceutical composition comprising a compound of formula (I) as described in any one of the embodiments of the invention or an isomer, prodrug, stable isotope derivative or pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, diluent, excipient and optionally further comprising one or more other RET kinase inhibitors.

The invention also relates to the use of a compound of formula (I) or an isomer, prodrug, stable isotope derivative or pharmaceutically acceptable salt thereof according to any one of the embodiments of the invention for the manufacture of a medicament for use as a RET kinase inhibitor.

The invention also relates to the use of a compound of formula (I) or an isomer, prodrug, stable isotope derivative or pharmaceutically acceptable salt thereof according to any of the embodiments of the invention for the manufacture of a medicament for the treatment or prevention of diseases mediated by RET kinase, such as cancer, in particular hematological malignancy, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain glioma.

The invention also relates to the use of a pharmaceutical composition according to the invention for the preparation of a medicament for the treatment or prevention of diseases mediated by RET kinase, such as cancer, in particular hematological malignancy, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain glioma.

The invention also relates to a method of treating or preventing a RET kinase mediated disease (e.g., a tumor, especially hematological malignancy, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, glioma) comprising administering to a patient in need thereof a therapeutically effective amount of a compound as described in any one of the embodiments of the invention or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described herein.

Another aspect of the invention relates to a compound as described in any one of the embodiments of the invention, or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, for use in the treatment or prophylaxis of a RET kinase mediated disease, such as a tumor, particularly hematological malignancy, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain glioma.

Another aspect of the invention relates to a pharmaceutical composition comprising a compound of formula (I) or an isomer, prodrug, stable isotope derivative or pharmaceutically acceptable salt thereof as described in any one of the embodiments of the invention and a pharmaceutically acceptable carrier, diluent, excipient for use in the treatment or prophylaxis of RET kinase mediated diseases, such as tumors, in particular hematological malignancies, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain glioma.

Another aspect of the present invention relates to a compound of formula (I) or a tautomer, a meso, a racemate, an enantiomer, a diastereomer, a mixture thereof, and a pharmaceutically acceptable salt thereof, as described in any one of the embodiments of the present invention for the treatment and/or prevention of a RET kinase mediated disease (e.g., tumor, etc.). The tumor is especially hematological malignancy, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain glioma.

Another aspect of the invention relates to a pharmaceutical composition comprising a compound of formula (I) or an isomer, prodrug, stable isotope derivative or a pharmaceutically acceptable salt thereof as described in any one of the embodiments of the invention and a pharmaceutically acceptable carrier, diluent, excipient, and optionally further comprising one or more other RET kinase inhibitors, as a treatment and/or prophylaxis of a RET kinase mediated disease (e.g., tumor, etc.).

According to the present invention, the drug may be any pharmaceutical dosage form including, but not limited to, tablets, capsules, solutions, lyophilized formulations, injections.

The pharmaceutical formulations of the present invention may be administered in dosage unit form containing a predetermined amount of active ingredient per dosage unit. Such units may contain, for example, from 0.5 mg to 1g, preferably from 1mg to 700 mg, particularly preferably from 5mg to 300 mg of a compound of the invention, or the pharmaceutical formulation may be administered in dosage unit form containing a predetermined amount of active ingredient per dosage unit, depending on the disorder being treated, the method of administration and the age, weight and condition of the patient. Preferred dosage unit formulations are those containing a daily dose or divided dose, or a corresponding fraction thereof, of the active ingredient as indicated above. In addition, this type of pharmaceutical formulation may be prepared using methods well known in the pharmaceutical arts.

The pharmaceutical formulations of the invention may be adapted for administration by any desired suitable method, for example by oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) methods. Such formulations may be prepared using all methods known in the pharmaceutical arts by, for example, combining the active ingredient with one or more excipients or one or more adjuvants.

Preparation method

The invention also provides a method for preparing the compound. The compounds of the present invention may be prepared by various methods well known in the art. For example, the compounds of the present invention can be synthesized as shown in schemes 1-10.

Scheme 1

R ¹⁶ is selected from C1-C4 alkyl;

P ¹ and P ² are each independently selected from halogen, such as chlorine, bromine, iodine, and the like;

the first step:

Adding a base (such as sodium bicarbonate) and R ¹⁶ OCOCl into a solvent selected from 1, 4-dioxane, tetrahydrofuran or water, and reacting at room temperature or under heating in an oil bath (20-50 ℃) to obtain a compound (II);

and a second step of:

Adding a compound (II), R ¹⁶ OH, diphenyl azide phosphate and alkali (such as anhydrous triethylamine) into anhydrous solvent (such as anhydrous toluene) in an inert gas (such as nitrogen or argon) atmosphere, heating in an oil bath, and keeping the temperature at 70-90 ℃ to perform Curtius rearrangement reaction to obtain a compound (III);

and a third step of:

adding a compound (IV), R ¹⁶ OH, diphenyl azide phosphate and alkali (such as anhydrous triethylamine) into anhydrous solvent (such as anhydrous toluene) in an inert gas (such as nitrogen or argon) atmosphere, heating in an oil bath, and keeping the temperature at 60-80 ℃ to perform Curtius rearrangement reaction to obtain a compound (III);

fourth step:

Adding a compound (III) and alkali (such as hexamethylenetetramine) into a solvent (such as trifluoroacetic acid and the like), heating in an oil bath, and maintaining the temperature at 80-100 ℃ for ring closure reaction to obtain a compound (V);

Fifth step:

dissolving the compound (V) in a solvent (such as ethanol and water), adding alkali (such as potassium hydroxide), heating in an oil bath, stirring for 1-3 hours at 70-90 ℃, adding an oxidant (such as potassium ferricyanide), and continuously stirring for 2-6 hours at the same temperature to perform hydrolysis and oxidation reaction to obtain a compound (VI);

Sixth step:

Dissolving the compound (VI) in 5-15% dilute hydrochloric acid, slowly adding an aqueous solution of sodium nitrite under ice bath, heating in oil bath, maintaining the temperature at 40-50 ℃ and stirring for 1-3 hours, then dropwise adding an aqueous solution of halogenated salt (such as potassium iodide and the like), continuously stirring for 1-3 hours at 40-50 ℃ and carrying out amino diazotization and halogenated reaction to obtain the compound (IX);

Seventh step:

Adding the compound (VII), lithium diisopropylamide and N, N-dimethylformamide into a solvent (such as tetrahydrofuran and the like), and carrying out formylation reaction at-78 ℃ to obtain a compound (VIII);

eighth step:

Adding the compound (VIII), potassium carbonate and methyl acetate into a solvent (such as anhydrous acetonitrile and the like), heating in an oil bath, and carrying out substitution ring closure reaction at 100-120 ℃ to obtain the compound (IX).

Scheme 2

P ¹、P²、P³ are each independently selected from halogen, such as chlorine, bromine, iodine, and the like;

PG ¹ is selected from t-butoxycarbonyl;

ar ¹ is selected from Wherein R ¹⁵ is selected from C1-C4 alkyl;

R ¹⁷ is selected from hydrogen, hydroxy;

R ¹⁸ is selected from hydrogen, amino or C1-C4 alkyl optionally substituted with one substituent selected from hydroxy, di (C1-C4 alkyl) amino, N-acetyl-N-methylamino, morpholin-4-yl, 1-ethylpiperazin-4-yl or 1-acetylpiperazin-4-yl;

r ¹⁹ is selected from C1-C4 alkoxy, optionally mono-or di-substituted phenyl, optionally mono-or di-substituted pyridinyl, 5 membered heteroaryl containing 1 or 2 heteroatoms selected from N, O, optionally substituted C1-C4 alkyl, optionally substituted 4-6 membered heterocyclyl containing 1 or 2 heteroatoms selected from N, O, C3-C6 cycloalkyl or amino optionally substituted by C1-C4 alkyl; the optional substituents are selected from halogen, hydroxy, C1-C4 alkoxy or C3-C6 cycloalkyl;

r ²⁰、R²¹ is each independently selected from hydrogen, C1-C4 alkyl;

the first step:

Dissolving a starting material (IX) (P ¹ is chlorine, P ² is bromine or iodine) and N-substituted pyrazole borate (or boric acid) in a solvent (dioxane and water), using a palladium complex such as tetra (triphenylphosphine) palladium as a catalyst, using sodium carbonate or potassium carbonate as a base, heating in an oil bath under the protection of nitrogen or argon, stirring at 70-100 ℃ for 2-12 hours, and carrying out Suzuki coupling reaction to obtain a compound (X);

and a second step of:

Dissolving a starting raw material (X), boric acid ester (or boric acid) (XI) in a solvent (dioxane or N, N-dimethylformamide and water), using a palladium complex such as tetrakis (triphenylphosphine) palladium as a catalyst, using potassium carbonate or potassium phosphate as a base, heating by an oil bath or microwaves under the protection of nitrogen or argon, stirring at 90-140 ℃ for 1-12 hours, and carrying out Suzuki coupling reaction to obtain a compound (XIV);

and a third step of:

Preparation of pinacol borate: dissolving a starting material (X) and a bisboronic acid pinacol ester in a solvent (such as dioxane), using a palladium complex such as 1,1' -bis (diphenylphosphino) ferrocene palladium dichloride as a catalyst or using tris (dibenzylideneacetone) dipalladium as a catalyst, adding a phosphine ligand such as tricyclohexylphosphine, using potassium acetate as a base, heating in an oil bath under the protection of nitrogen or argon, and stirring at the temperature of 60-100 ℃ for 2-12 hours to obtain a compound (XII);

fourth step:

Dissolving a starting material (XII) and a heterocyclic aryl halide (XIII) in a solvent (dioxane or N, N-dimethylformamide and water), using a palladium complex such as tetrakis (triphenylphosphine) palladium as a catalyst, using potassium carbonate or potassium phosphate as a base, heating in an oil bath or a microwave under the protection of nitrogen or argon, stirring at 90-140 ℃ for 1-12 hours, and carrying out a Suzuki coupling reaction to obtain a compound (XIV);

Fifth step:

Deprotection of PG ¹, for example of t-butoxycarbonyl, uses trifluoroacetic acid or hydrochloric acid as acid; in a solvent such as methylene dichloride or dioxane, the reaction is carried out at the temperature of 0-25 ℃; obtaining a compound (XV) through reaction;

Sixth step:

Dissolving the initial raw material (XV) and corresponding acyl chloride in a solvent (such as tetrahydrofuran, methylene dichloride and the like), adding alkali such as triethylamine and the like, stirring at room temperature for 20 minutes to 18 hours for condensation; or dissolving the initial raw material (XV) and the corresponding carboxylic acid in a solvent (tetrahydrofuran, dichloromethane or N, N-dimethylformamide and the like), adding a condensing agent such as 2- (7-benzotriazol-N, N, N ', N' -tetramethylurea hexafluorophosphate and the like, adding a base such as triethylamine and the like, stirring at room temperature for 20 minutes to 18 hours for condensation; compound (XVI) can be obtained;

Seventh step:

Dissolving the initial raw material (XV) and the corresponding isocyanate in a solvent (tetrahydrofuran, methylene dichloride or N, N-dimethylformamide and the like), and stirring at room temperature for 20 minutes to 18 hours; or the initial raw material (XV) and N, N '-carbonyl diimidazole or N, N' -carbonyl di (1, 2, 4-triazole) are dissolved in solvent (tetrahydrofuran, methylene dichloride or N, N-dimethylformamide, etc.), stirred for 20 minutes at room temperature or in oil bath heating (20-70 ℃), corresponding amine is added, stirred for 20 minutes to 18 hours at room temperature or in oil bath heating (20-70 ℃), and condensed; compound (XVII) can be obtained;

eighth step:

Dissolving a starting material (IX) (P ¹ is bromine or iodine, P ² is chlorine) and boric acid ester (or boric acid) (XI) in a solvent (dioxane and water), using palladium complex such as tetra (triphenylphosphine) palladium as a catalyst, using sodium carbonate or potassium carbonate as a base, heating in an oil bath under the protection of nitrogen or argon, stirring at 50-100 ℃ for 2-12 hours, and carrying out Suzuki coupling reaction to obtain a compound (XVIII);

ninth step:

The starting material (XVIII), N-substituted pyrazole borate (or boric acid) is dissolved in a solvent (dioxane or N, N-dimethylformamide and water), palladium complex such as tetrakis (triphenylphosphine) palladium is used as a catalyst, potassium carbonate or potassium phosphate is used as a base, and under the protection of nitrogen or argon, the mixture is heated by an oil bath or microwaves, stirred for 1 to 12 hours at the temperature of 90 to 140 ℃ and subjected to Suzuki coupling reaction to obtain the compound (XIV).

Scheme 3:

the first step:

Dissolving 8-aminoquinoline (XIX) in dichloromethane, sequentially adding triethylamine and benzoyl chloride, and reacting at 25 ℃ for 1 hour after the addition to obtain N- (quinoline-8-yl) benzamide (XX);

and a second step of:

N- (quinolin-8-yl) benzamide (XX) was added to water, N-bromosuccinimide was added in portions, and the mixture was reacted at 25℃for 15 hours after the addition. To give N- (5-bromoquinolin-8-yl) benzamide (XXI);

and a third step of:

Dissolving N- (5-bromoquinolin-8-yl) benzamide (XXI) in ethanol, adding a 1M sodium hydroxide solution in ethanol with stirring at 25℃and stirring at 85℃for 15 hours to give 5-bromo-8-aminoquinoline (XXII);

Fourth step:

dissolving 5-bromo-8-aminoquinoline (XXII) in acetonitrile, adding N-chlorosuccinimide, and stirring at 80 ℃ for 6 hours to obtain 5-bromo-7-chloro-8-aminoquinoline (XXIII);

Fifth step:

5-bromo-7-chloro-8-aminoquinoline (XXIII) was added to water, then concentrated sulfuric acid was added dropwise at 0℃until most of the solid disappeared, 4M aqueous sodium nitrite was added dropwise when the solution turned orange-red, stirred at 0℃for 0.5 hours, finally this mixture was added dropwise to hypophosphorous acid, stirred at 65℃for 4 hours, and the mixture was poured into saturated aqueous sodium hydroxide solution (3L) to give 5-bromo-7-chloroquinoline (XXIV).

Scheme 4

P ³ is selected from halogen, such as chlorine, bromine, iodine, etc.;

PG ¹ is selected from t-butoxycarbonyl;

R ¹⁵ is selected from C1-C4 alkyl;

R ¹⁷ is selected from hydrogen, hydroxy;

R ¹⁸ is selected from amino or C1-C4 alkyl, wherein said alkyl is optionally substituted with one substituent selected from hydroxy, di (C1-C4 alkyl) amino, N-acetyl-N-methylamino, morpholin-4-yl, 1-ethylpiperazin-4-yl or 1-acetylpiperazin-4-yl;

the first step:

Dissolving a starting material (XXIV) and boric acid ester (or boric acid) (XI) in a solvent (dioxane and water), using a palladium complex such as tetra (triphenylphosphine) palladium as a catalyst, using sodium carbonate or potassium carbonate as a base, heating in an oil bath under the protection of nitrogen or argon, stirring at 50-100 ℃ for 2-12 hours, and carrying out Suzuki coupling reaction to obtain a compound (XXV);

and a second step of:

Dissolving a starting material (XXV) and N-substituted pyrazole borate (or boric acid) in a solvent (dioxane or N, N-dimethylformamide and water), using a palladium complex such as tetrakis (triphenylphosphine) palladium as a catalyst, using potassium carbonate or potassium phosphate as a base, heating in an oil bath or a microwave under the protection of nitrogen or argon, stirring at 90-140 ℃ for 1-12 hours, and carrying out Suzuki coupling reaction to obtain a compound (XXVI);

and a third step of:

Deprotection of t-butoxycarbonyl group uses trifluoroacetic acid or hydrochloric acid as acid; deprotection of PG ¹ in a solvent such as methylene chloride or dioxane, and reaction at 0 to 25℃to give compound (XXVII);

fourth step:

Dissolving the initial raw material (XXVII) and corresponding acyl chloride in a solvent (such as tetrahydrofuran, methylene dichloride and the like), adding alkali such as triethylamine and the like, stirring at room temperature for 20 minutes to 18 hours for condensation; or dissolving the starting material (XXVII) and the corresponding carboxylic acid in a solvent (tetrahydrofuran, methylene dichloride or N, N-dimethylformamide and the like), adding a condensing agent such as 2- (7-benzotriazol-N, N, N ', N' -tetramethylurea hexafluorophosphate and the like, adding a base such as triethylamine and the like, stirring at room temperature for 20 minutes to 18 hours for condensation; compound (XXVIII) can be obtained;

Fifth step:

Dissolving a starting material (XXIV) and a bisboronic acid pinacol ester in tetrahydrofuran, adding potassium acetate and 1,1' -bis (diphenylphosphino) ferrocene palladium dichloride, bubbling the system with nitrogen for 10 minutes, stirring at 60 ℃ for 2 hours under the protection of nitrogen, and directly performing rotary evaporation to obtain 7-chloro-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) quinoline (XXIX), wherein the crude product is used in the next step without purification;

Sixth step:

The starting material 7-chloro-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) quinoline (XXIX), a heterocyclic aryl halide (XIII) is dissolved in a solvent (dioxane or N, N-dimethylformamide and water), palladium complex such as tetrakis (triphenylphosphine) palladium is used as a catalyst, potassium carbonate or potassium phosphate is used as a base, and the mixture is heated in an oil bath or microwave under the protection of nitrogen or argon, stirred at a temperature of 90-140 ℃ for 1-12 hours, and subjected to Suzuki coupling reaction to obtain the compound (XXV).

Scheme 5

P ³ is selected from halogen, such as chlorine, bromine, iodine, etc.;

PG ¹ is selected from t-butoxycarbonyl;

R ¹⁷ is selected from hydrogen, hydroxy;

the first step:

Dissolving 2-chloro-4-ethoxy-6-fluorobenzaldehyde (XXX) in dimethyl sulfoxide, adding potassium carbonate and formamidine acetate under stirring, and stirring at 110 ℃ for 6 hours to obtain 5-chloro-7-ethoxyquinazoline (XXXI);

and a second step of:

Dissolving a starting material of 5-chloro-7-ethoxyquinazoline (XXXI), boric acid ester (or boric acid) (XI) in a solvent (dioxane or N, N-dimethylformamide and water), using a palladium complex such as tetrakis (triphenylphosphine) palladium as a catalyst, using potassium carbonate or potassium phosphate as a base, heating in an oil bath or a microwave under the protection of nitrogen or argon, stirring at 90-140 ℃ for 1-12 hours, and carrying out Suzuki coupling reaction to obtain a compound (XXXIII);

and a third step of:

Dissolving a starting material of 5-chloro-7-ethoxyquinazoline (XXXI) and bisboronic acid pinacol ester in a solvent (such as dioxane), using a palladium complex such as 1,1' -bis (diphenylphosphino) ferrocene palladium dichloride as a catalyst or using tris (dibenzylideneacetone) dipalladium as a catalyst, adding a phosphine ligand such as tricyclohexylphosphine, using potassium acetate as a base, heating in an oil bath under the protection of nitrogen or argon, and stirring at the temperature of 60-100 ℃ for 2-12 hours to obtain a compound (XXXII);

fourth step:

Dissolving a starting material (XXXII) and a heterocyclic aryl halide (XIII) in a solvent (dioxane or N, N-dimethylformamide and water), using a palladium complex such as tetrakis (triphenylphosphine) palladium as a catalyst, using potassium carbonate or potassium phosphate as a base, heating in an oil bath or a microwave under the protection of nitrogen or argon, stirring at 90-140 ℃ for 1-12 hours, and carrying out a Suzuki coupling reaction to obtain a compound (XXXIII);

Fifth step:

Deprotection of PG ¹, for example of t-butoxycarbonyl, uses trifluoroacetic acid or hydrochloric acid as acid; in a solvent such as methylene dichloride or dioxane, the reaction is carried out at the temperature of 0-25 ℃; the reaction gives compound (XXXIV);

Sixth step:

Dissolving the starting material (XXXIV) and the corresponding carboxylic acid in a solvent (tetrahydrofuran, dichloromethane or N, N-dimethylformamide and the like), adding a condensing agent such as 2- (7-benzotriazol-N, N, N ', N' -tetramethylurea hexafluorophosphate and the like, adding a base such as triethylamine and the like, stirring at room temperature for 20 minutes to 18 hours for condensation; compound (XXXV) can be obtained.

Scheme 6

PG ¹ is selected from t-butoxycarbonyl;

R ¹⁷ is selected from hydrogen, hydroxy;

The first step: dissolving a starting material of 5, 7-dichloro-1, 6-naphthyridine (synthetic reference: PCT Int. Appl., 2011134971) (XXXVI), boric acid ester (or boric acid) (XI) in a solvent (dioxane or N, N-dimethylformamide and water), using a palladium complex such as tetrakis (triphenylphosphine) palladium as a catalyst or tris (dibenzylideneacetone) dipalladium as a catalyst, using tricyclohexylphosphine as a ligand, using potassium carbonate or potassium phosphate as a base, heating with an oil bath or microwaves under the protection of nitrogen or argon, and stirring at 80-90 ℃ for 12-16 hours, and performing Suzuki coupling reaction to obtain a compound (XXXVII);

and a second step of:

Dissolving a starting material (XXXVII), N-methylpyrazole borate (or boric acid) in a solvent (dioxane or N, N-dimethylformamide and water), using a palladium complex such as tetrakis (triphenylphosphine) palladium as a catalyst, using potassium carbonate or potassium phosphate as a base, heating in an oil bath or a microwave under the protection of nitrogen or argon, stirring at 120-140 ℃ for 1-12 hours, and carrying out Suzuki coupling reaction to obtain a compound (XXXVIII);

and a third step of:

Deprotection of PG ¹, for example, deprotection of t-butoxycarbonyl group uses hydrogen chloride dioxane solution or ethyl hydrogen chloride acetate solution as acid; in solvents such as methanol, ethyl acetate or dioxane, the reaction is carried out at the temperature of 0-25 ℃;

fourth step:

Dissolving the starting material (XXXIX) and the corresponding carboxylic acid in a solvent (tetrahydrofuran, dichloromethane or N, N-dimethylformamide and the like), adding a condensing agent such as 2- (7-benzotriazol-N, N, N ', N' -tetramethylurea hexafluorophosphate and the like, adding a base such as triethylamine and the like, stirring at room temperature for 20 minutes to 18 hours for condensation; compound (XXXX) is obtained.

Scheme 7

PG ¹ is selected from t-butoxycarbonyl;

r ¹⁴ is selected from C1-C4 alkyl or C1-C4 alkyl optionally substituted with 1-methylpiperidin-4-yl;

R ¹⁷ is selected from hydrogen, hydroxy;

the first step:

starting material XXXVII (R ¹⁴ is ethyl) was dissolved in sodium ethoxide ethanol solution prepared using sodium and ethanol, heated in an oil bath and stirred at 100℃for about 16 hours to give compound (XXXXI); or (b)

Dissolving a starting material XXXVII and 1-methyl-4-piperidinemethanol in 1, 4-dioxane, using a metal complex such as tris (dibenzylideneacetone) dipalladium as a catalyst and cesium carbonate as a base, bubbling the reaction solution with nitrogen for ten minutes, and heating and stirring the reaction solution with a microwave at 110 ℃ under the protection of nitrogen for 1 hour to obtain a compound (XXXXI);

and a second step of:

deprotection of PG ¹, for example, deprotection of t-butoxycarbonyl uses ethyl chloride solution as acid; in ethyl acetate and other solvents, the reaction is carried out at 0-25 ℃;

and a third step of:

dissolving the starting material (XXXXII) and the corresponding carboxylic acid in a solvent (tetrahydrofuran, dichloromethane or N, N-dimethylformamide, etc.), adding a condensing agent such as 2- (7-benzotriazol-N, N, N ', N' -tetramethylurea hexafluorophosphate, etc., adding a base such as triethylamine, etc., stirring at room temperature for 20 minutes to 18 hours for condensation; compound (XXXXIII) can be obtained.

Scheme 8

PG ¹ is selected from t-butoxycarbonyl;

r ²² is selected from phenyl optionally mono-or disubstituted, pyridinyl optionally mono-or disubstituted, said substituents being selected from halogen, C1-C4 alkoxy;

r ²³ is selected from C1-C4 alkyl;

Selected from:

the first step:

Dissolving a starting material of 5, 7-dichloro-1, 6-naphthyridine (synthetic reference: PCT Int. Appl., 2011134971) (XXXVI) and boric acid ester (or boric acid) (XXXXIV) in a solvent (dioxane or N, N-dimethylformamide and water), using a palladium complex such as tetrakis (triphenylphosphine) palladium as a catalyst or tris (dibenzylideneacetone) dipalladium as a catalyst, using tricyclohexylphosphine as a ligand, using potassium carbonate or potassium phosphate as a base, heating by an oil bath or microwaves under the protection of nitrogen or argon, and stirring at 80-90 ℃ for 12-16 hours, and performing Suzuki coupling reaction to obtain a compound (XXXXXXV);

and a second step of:

Dissolving a starting material (XXXXV) and N-methylpyrazole borate (or boric acid) in a solvent (dioxane or N, N-dimethylformamide and water), using a palladium complex such as tetrakis (triphenylphosphine) palladium as a catalyst, using potassium phosphate as a base, heating in an oil bath or under the protection of nitrogen or argon, stirring at 120-140 ℃ for 1-12 hours, and carrying out Suzuki coupling reaction to obtain a compound (XXXXVI);

and a third step of:

Deprotection of PG ¹, for example, deprotection of t-butoxycarbonyl group uses hydrogen chloride dioxane solution or ethyl hydrogen chloride acetate solution as acid; in solvents such as methanol, ethyl acetate or dioxane, the reaction is carried out at 0-25 ℃ to obtain a compound (XXXXVII);

fourth step:

The starting material (XXXXVII) and the corresponding aldehyde or ketone are dissolved in a solvent (methanol, ethanol or 1, 2-dichloroethane, etc.), and a reducing agent such as sodium triacetoxyborohydride, etc. is added thereto, and the mixture is heated and stirred at 20 to 50℃for 12 to 48 hours at room temperature or in an oil bath to obtain the compound (XXXXVIII).

Fifth step:

dissolving the starting material (XXXXVII) and the corresponding carboxylic acid in a solvent (tetrahydrofuran, dichloromethane or N, N-dimethylformamide, etc.), adding a condensing agent such as 2- (7-benzotriazol-N, N, N ', N' -tetramethylurea hexafluorophosphate, etc., adding a base such as triethylamine, etc., stirring at room temperature for 20 minutes to 18 hours for condensation; compound (XXXXIX) can be obtained.

Sixth step:

Dissolving the starting material (XXXXVII) and the corresponding carboxylic acid in a solvent (N, N-dimethylformamide, etc.), adding alkylsulfonyl chloride R ²³S(O)₂ Cl, adding a base such as triethylamine, etc., and stirring at room temperature for 15 minutes to 18 hours; compound (XXXXX) is obtained.

Scheme 9

R ²³ is selected from C1-C4 alkyl;

the first step:

dissolving a starting material of 5, 7-dichloro-1, 6-naphthyridine (synthetic reference: PCT Int. Appl., 2011134971) (XXXVI) and boric acid ester (or boric acid) (XXXXXI) in a solvent (dioxane or N, N-dimethylformamide and water), using a palladium complex such as tetrakis (triphenylphosphine) palladium as a catalyst or tris (dibenzylideneacetone) dipalladium as a catalyst, using tricyclohexylphosphine as a ligand, using potassium carbonate or potassium phosphate as a base, stirring for 12-16 hours at 80-90 ℃ under the protection of nitrogen or argon, and carrying out a Suzuki coupling reaction to obtain a compound (XXXXXII);

and a second step of:

The starting material (XXXXXII), N-methylpyrazole borate (or boric acid) is dissolved in a solvent (dioxane or N, N-dimethylformamide and water), palladium complex such as tetra (triphenylphosphine) palladium is used as a catalyst, potassium phosphate is used as a base, and stirring is carried out for 1-12 hours at 120-140 ℃ under the protection of nitrogen or argon, and Suzuki coupling reaction is carried out to obtain a compound (XXXXXIII).

Scheme 10

R ¹⁷ is selected from hydrogen, hydroxy or amino;

the first step:

dissolving the initial raw material (XXXXXIV) in N, N-dimethylformamide, adding sodium hydrogen, stirring for half an hour at room temperature, then adding 2-fluoropyridine, and stirring for 4-16 hours at 80-90 ℃ to obtain a compound (XXXXXV);

and a second step of:

dissolving a starting material (XXXXXV), 5-bromo-2-fluoropyridine and N, N-diisopropylethylamine in dimethyl sulfoxide, and stirring at 90-110 ℃ for 10-20 hours to obtain a compound (XXXXXVI);

and a third step of:

Dissolving a starting material (XXXXXVI), pinacol ester, potassium acetate and 1,1' -bis-diphenylphosphino ferrocene palladium dichloride in dioxane under the protection of nitrogen, stirring for 4-10 hours at 80-100 ℃, cooling the obtained mixture to room temperature, adding 5, 7-dichloro-1, 6-naphthyridine (synthetic reference: PCT Int. Appl., 2011134971) (XXXVI), palladium complex such as tetra (triphenylphosphine) palladium serving as a catalyst or tris (dibenzylideneacetone) dipalladium serving as a catalyst, adding water, stirring for 12-20 hours at 80-90 ℃ and performing Suzuki coupling reaction to obtain a compound (XXXXXVII);

fourth step:

The starting material (XXXXXVII), N-methylpyrazole borate (or boric acid) is dissolved in a solvent (dioxane or N, N-dimethylformamide and water), palladium complex such as tetrakis (triphenylphosphine) palladium is used as a catalyst, potassium phosphate is used as a base, and stirring is carried out for 2-12 hours at 130-150 ℃ under the protection of nitrogen or argon, so that Suzuki coupling reaction is carried out, and a compound (XXXXXVIII) is obtained.

Detailed Description

Definition of the definition

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

The expression "Cx-Cy" as used in the present invention means a range of carbon atom numbers, wherein x and y are both integers, for example, C3-C8 cycloalkyl represents cycloalkyl having 3 to 8 carbon atoms, -C0-C2 alkyl represents alkyl having 0 to 2 carbon atoms, wherein-C0 alkyl means a chemical single bond.

In the present invention, the term "alkyl" refers to saturated aliphatic hydrocarbon groups, including straight and branched chain groups of 1 to 20 carbon atoms, which may be, for example, straight and branched chain groups of 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, and various branched isomers thereof, and the like. The alkyl group may be optionally substituted or unsubstituted.

In the present invention, the term "alkenyl" refers to straight-chain, branched-chain hydrocarbon groups containing at least 1 carbon-carbon double bond, which may include from 2 to 20 carbon atoms, for example, straight-chain and branched-chain groups which may be from 2 to 18 carbon atoms, from 2 to 12 carbon atoms, from 2 to 8 carbon atoms, from 2 to 6 carbon atoms, or from 2 to 4 carbon atoms. Wherein 1-3 carbon-carbon double bonds may be present, preferably 1 carbon-carbon double bond is present. The term "C2-C4 alkenyl" refers to alkenyl groups having 2 to 4 carbon atoms. Including ethenyl, propenyl, butenyl, buten-2-yl, 2-methylbutenyl. Alkenyl groups may be optionally substituted or unsubstituted.

In the present invention, the term "alkynyl" refers to straight-chain, branched-chain hydrocarbon groups containing at least 1 carbon-carbon triple bond, which may include from 2 to 20 carbon atoms, for example, straight-chain and branched-chain groups which may be from 2 to 18 carbon atoms, from 2 to 12 carbon atoms, from 2 to 8 carbon atoms, from 2 to 6 carbon atoms, or from 2 to 4 carbon atoms. Wherein 1-3 carbon-carbon triple bonds may be present, preferably 1 carbon-carbon triple bond. The term "C2-C4 alkynyl" refers to alkynyl groups having 2 to 4 carbon atoms. Non-limiting examples include ethynyl, propynyl, butynyl and butyn-2-yl, 3-methylbutynyl. Alkynyl groups may be optionally substituted or unsubstituted.

In the present invention, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic cyclic hydrocarbon group comprising 3 to 12 ring atoms, which may be, for example, 3 to 12, 3 to 10, 3 to 8 or 3 to 6 ring atoms, or may be a 3, 4, 5, 6 membered ring. Non-limiting examples of monocyclic ring groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. Cycloalkyl groups may be optionally substituted or unsubstituted.

In the present invention, the term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon group comprising 3 to 20 ring atoms, which may be, for example, 3 to 16, 3 to 12, 3 to 10, 3 to 8 or 3 to 6 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen or heteroatoms of S (O) m (where m is an integer from 0 to 2), but excluding the ring portion of-O-, -O-S-or-S-, the remaining ring atoms being carbon. Preferably 3 to 12 ring atoms, of which 1 to 4 are heteroatoms, more preferably the heterocyclyl ring contains 3 to 10 ring atoms, more preferably 3 to 8 ring atoms, most preferably 5-membered ring or 6-membered ring, of which 1 to 4 are heteroatoms, more preferably 1 to 3 are heteroatoms, most preferably 1 to 2 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Polycyclic heterocyclyl groups include spiro, fused and bridged heterocyclic groups. The heterocyclyl group may be optionally substituted or unsubstituted.

In the present invention, the term "heterocyclylene" refers to a substituted or unsubstituted heterocyclic group having two terminal monovalent radical cores, resulting from the removal of one hydrogen atom from each of the two terminal atoms; the heterocyclic group has the meaning as described above. Non-limiting examples of "heterocyclylene" include pyrrolidinylene, piperidinyl, piperazinylene, morpholinylene, and the like.

In the present invention, the term "spiroheterocyclyl" refers to a 5 to 20 membered, monocyclic, polycyclic heterocyclic group sharing one atom (referred to as the spiro atom) between the monocyclic rings, wherein one or more of the ring atoms is selected from nitrogen, oxygen or a heteroatom of S (O) m (where m is an integer from 0 to 2) and the remaining ring atoms are carbon. These may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system. Preferably 6 to 14 membered, more preferably 7 to 10 membered. The spirocyclic groups are classified into single-, double-or multiple-spiroheterocyclic groups according to the number of common spiro atoms between rings, preferably Shan Luohuan groups and double-spirocyclic groups. More preferably 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered single spiro-cyclic group. Non-limiting examples of spiro radicals include

. The spirocyclic group may be optionally substituted or unsubstituted.

In the present invention, the term "spiroheterocyclyl" refers to a substituted or unsubstituted spiroheterocyclyl having two terminal monovalent radical cores, resulting from the removal of one hydrogen atom from each of the two terminal atoms; the spiroheterocyclyl has the meaning described above. Non-limiting examples of "spiroylene" radicals include。

In the present invention, the term "fused heterocyclyl" refers to a 5 to 20 membered, polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with the other rings in the system, one or more of which may contain one or more double bonds, but none of which has a fully conjugated pi electron system in which one or more ring atoms are selected from nitrogen, oxygen or a heteroatom of S (O) m (where m is an integer from 0 to 2) and the remaining ring atoms are carbon. Preferably 6 to 14 membered, more preferably 7 to 10 membered. The number of constituent rings may be classified into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic, more preferably a 4-membered/5-membered, 5-membered/6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclyl groups include

。

The heterocyclyl ring may be fused to an aryl, heteroaryl or cyclyl ring, wherein the ring attached to the parent structure is heterocyclyl, non-limiting examples include:

Etc. The fused heterocyclic group may be optionally substituted or unsubstituted.

In the present invention, the term "fused heterocyclic group" refers to a substituted or unsubstituted fused heterocyclic group having two terminal monovalent group cores, which is produced by removing one hydrogen atom from each of the two terminal atoms; the fused heterocyclic group has the meaning as described above. Non-limiting examples of "fused heterocyclylene" include

。

In the present invention, the term "aryl" refers to a6 to 14 membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group, a polycyclic (i.e., rings bearing adjacent pairs of carbon atoms) group having a conjugated pi-electron system, preferably 6 to 10 membered, such as phenyl and naphthyl, most preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cyclyl ring, wherein the ring attached to the parent structure is an aryl ring, non-limiting examples of which include:

Aryl groups may be substituted or unsubstituted.

In the present invention, the term "heteroaryl" refers to heteroaromatic systems containing from 1 to 4 heteroatoms, including oxygen, sulfur and nitrogen, from 5 to 14 ring atoms. Preferably 5 to 10 yuan. More preferably heteroaryl is 5-or 6-membered, such as furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, oxazolyl, isoxazolyl, and the like, which heteroaryl ring may be fused to an aryl, heterocyclyl, or cyclyl ring, wherein the ring attached to the parent structure is a heteroaryl ring, non-limiting examples include:

。

heteroaryl groups may be optionally substituted or unsubstituted.

In the present invention, the term "halogen" refers to fluorine, chlorine, bromine or iodine.

In the present invention, the term "cyano" refers to-CN.

In the present invention, the term "nitro" refers to the NO ₂ group.

In the present invention, the term "hydroxy" refers to an-OH group.

In the present invention, the term "amino" refers to the-NH ₂ group. Substitution of one or two hydrogens on the amino group with an alkyl group having the meaning set forth above may form a mono-or dialkylamino group.

In the present invention, the term "carbonyl" refers toA group. One side of the carbon on the carbonyl group is attached to an alkyl group, which has the meaning as previously described, to form an alkylcarbonyl group.

In the present invention, the term "alkoxy" refers to an alkyl group attached through an oxygen bridge, including alkyloxy, cycloalkyloxy, and heterocycloalkyloxy. The alkyl, heterocycloalkyl and cycloalkyl have the meanings indicated above.

In the present invention, the term "heteroaryloxy" refers to a heteroaryl group linked through an oxygen bridge, said heteroaryl group having the meaning as described hereinbefore.

In the present invention, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxyl group, and includes hydroxyalkyl, hydroxycycloalkyl, and hydroxyheterocycloalkyl groups. The alkyl, heterocycloalkyl and cycloalkyl have the meanings indicated above.

In the present invention, the term "haloalkyl" refers to an alkyl substituent in which at least one hydrogen is replaced by a halogen group. Typical halogen groups include chlorine, fluorine, bromine and iodine. Examples of haloalkyl include fluoromethyl, fluoroethyl, chloromethyl, chloroethyl, 1-bromoethyl, difluoromethyl, trifluoromethyl and 1, 1-trifluoroethyl. It will be appreciated that if a substituent is substituted with more than one halogen group, those halogen groups may be the same or different (unless otherwise indicated).

In the present invention, "optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs or does not. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may be, but is not necessarily, present, and the description includes cases where the heterocyclic group is substituted with an alkyl group and cases where the heterocyclic group is not substituted with an alkyl group.

In the present invention, "substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable when bound to carbon atoms having unsaturated (e.g., olefinic) bonds.

Such substituents include, but are not limited to, the various groups described previously.

The claimed compounds include not only the compounds themselves, but also isomers, prodrugs, stable isotopic derivatives of the compounds or pharmaceutically acceptable salts thereof.

The term "pharmaceutical composition" as used herein means a mixture containing one or more of the isomers, prodrugs, stable isotopic derivatives of the compounds of the present invention or pharmaceutically acceptable salts thereof, and other chemical components. Other components such as pharmaceutically acceptable carriers, diluents and excipients. The purpose of the pharmaceutical composition is to promote the administration to organisms, facilitate the absorption of active ingredients and thus exert biological activity.

The term "comprising" when used in the specification is comprised of ….

The term "room temperature" as used herein means 15-30deg.C.

The "stable isotope derivatives" of the present invention include: isotopically substituted derivatives in which any hydrogen atom in formula I is substituted with 1 to 5 deuterium atoms, isotopically substituted derivatives in which any carbon atom in formula I is substituted with 1 to 3 carbon 14 atoms, or isotopically substituted derivatives in which any oxygen atom in formula I is substituted with 1 to 3 oxygen 18 atoms.

The "pharmaceutically acceptable salts" of the present invention are discussed in Berge, et al, "Pharmaceutically acceptable salts," j.pharm.sci., 66, 1-19 (1977) and are readily apparent to pharmaceutical chemists that are substantially non-toxic and provide desirable pharmacokinetic properties, palatability, absorption, distribution, metabolism, or excretion, and the like.

Pharmaceutically acceptable salts of the invention can be synthesized by general chemical methods.

In general, salts can be prepared by reacting the free base or acid with an equivalent stoichiometric or excess of an acid (inorganic or organic) or base in a suitable solvent or solvent composition.

The term "prodrug" as used herein refers to a compound that is converted to the original active compound after metabolism in vivo. Typically, prodrugs are inactive substances or less active than the active parent compound, but may provide ease of handling, administration or improved metabolic characteristics.

The term "isomer" as used herein refers to the tautomers, meso, racemates, enantiomers, diastereomers, mixtures thereof, and the like, of the compounds of formula (I) according to the invention. All such isomers, including stereoisomers, geometric isomers are encompassed by the present invention. The geometric isomers include cis and trans isomers.

The invention includes any polymorph of the compound or salt thereof, and any hydrate or other solvate.

In the present invention, the term "patient" generally refers to a mammal, especially a human.

In the present invention, the term "tumor" includes benign tumors and malignant tumors, such as cancers.

In the present invention, the term "cancer" includes RET kinase mediated tumors including, but not limited to, hematological malignancies, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain glioma.

In the present invention, the term "therapeutically effective amount" is meant to include an amount of a compound of the present invention that is effective to treat or prevent a related disorder mediated by RET kinase.

Examples

The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.

The structures of all compounds of the invention can be identified by nuclear magnetic resonance (¹ H NMR) and/or Mass Spectrometry (MS).

¹ H NMR chemical shifts (δ) are reported in PPM (parts per million parts per million). NMR was performed by Bruker AVANCE III-400MHz spectrometer. Suitable solvents are selected from deuterated chloroform (CDCl ₃), deuterated methanol (CD ₃ OD), deuterated dimethyl sulfoxide (DMSO-d ₆), and the like, tetramethylsilane as an internal standard (TMS).

Low resolution Mass Spectrometry (MS) was determined by an Agilent 1260 HPLC/6120 mass spectrometer using Agilent ZORBAX XDB-C18, 4.6X10 mm,3.5 μm.

Gradient elution condition one: 0:95% solvent A1 and 5% solvent B1, 1-2:5% solvent A1 and 95% solvent B1;2.01-2.50, 95% solvent A1 and 5% solvent B1. The percentage is the volume percentage of a certain solvent to the total solvent volume. Solvent A1:0.01% formic acid aqueous solution; solvent B1:0.01% formic acid in acetonitrile; the percentage is the volume percentage of solute in the solution.

The thin-layer silica gel plate is a tobacco stand yellow sea HSGF254 or Qingdao GF254 silica gel plate. Column chromatography generally uses 100-200 or 200-300 mesh silica gel of yellow sea as carrier.

Preparative liquid chromatography (prep-HPLC) using WATERS SQD mass spectrometry directed to a high pressure liquid chromatography separator, XBridge-C18; 30X 150 mm preparation column, 5 μm;

the method comprises the following steps: acetonitrile-water (0.2% formic acid), flow rate 25 mL/min; the second method is as follows: acetonitrile-water (0.8% ammonium bicarbonate), flow rate 25 mL/min;

The known starting materials of the present invention may be synthesized using or according to methods known in the art, or may be purchased from Acros Organics, ALDRICH CHEMICAL Company, shaoshan chemical technology (Accela ChemBio Inc), shanghai Pi De medicine, shanghai Ala Ding Huaxue, shanghai Michelson chemistry, carbofuran chemistry, an Naiji chemistry, and the like.

In the examples, if no special description exists, the solvent used in the reaction is anhydrous solvent, wherein the anhydrous tetrahydrofuran uses commercial tetrahydrofuran, sodium block is used as a water scavenger, benzophenone is used as an indicator, reflux is carried out to the solution under the protection of argon gas to form bluish violet, distillation and collection are carried out, the solution is stored at room temperature under the protection of argon gas, other anhydrous solvents are purchased from Angust chemical and carbofuran chemical, and all transfer and use of the anhydrous solvents are carried out under the protection of argon gas if no special description exists.

In the examples, unless otherwise specified, the reaction was carried out under an argon atmosphere or a nitrogen atmosphere.

An argon or nitrogen atmosphere means that the reactor flask is connected to a balloon of argon or nitrogen of about 1L volumes.

The hydrogen atmosphere is defined as the reaction flask being connected to a balloon of hydrogen gas of about 1L volumes.

The hydrogenation reaction is usually vacuumized, filled with hydrogen and repeatedly operated for 3 times.

In the examples, the reaction temperature was room temperature and the temperature range was 15 ℃to 30℃unless otherwise specified.

The progress of the reaction in the examples was monitored by Thin Layer Chromatography (TLC) using a system of developing agents of a: methylene chloride and methanol systems; b: petroleum ether and ethyl acetate systems. The volume ratio of the solvent is adjusted according to the polarity of the compound.

The system of eluent for column chromatography and the system of developing agent for thin layer chromatography used for purifying the compound include a: methylene chloride and methanol systems; b: petroleum ether and ethyl acetate systems. The volume ratio of the solvent is adjusted according to the polarity of the compound, and can be adjusted by adding a small amount of triethylamine, an acidic or alkaline reagent and the like.

Example 23

2-Chloro-N- ((3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-yl) benzamide

First step

((3S, 4S) -1- (5-bromopyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamic acid tert-butyl ester

The compound tert-butyl 2-fluoro-5-bromopyridine (1.08 g,5.00 mmol), ((3S, 4S) -3-hydroxypiperidin-4-yl) carbamate (synthetic reference: WO 2004058144A 2) (1.18 g,5.00 mmol) and N, N-diisopropylethylamine (1.29 g,10.00 mmol) were dissolved in dimethyl sulfoxide (10 mL) and heated to 90℃for overnight reaction. After cooling, the reaction mixture was diluted with ethyl acetate (100 mL), the organic phase was washed with water (100 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel (ethyl acetate/petroleum ether=2:3) to give the desired product tert-butyl (3 s,4 s) -1- (5-bromopyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamate (1.40 g, yellow solid). Yield: 75.4 Percent of the total weight of the composition. MS M/z (ESI) 372&374 [ M+1 ];

Second step

((3S, 4S) -1- (5- (7-chloroquinolin-5-yl) pyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamic acid tert-butyl ester

The compound ((3S, 4S) -1- (5-bromopyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamic acid tert-butyl ester (0.15 g,0.54 mmol), 7-chloro-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) quinoline (synthetic reference: WO 2017032840A 1) (0.15 g,0.54 mmol), tetrakis (triphenylphosphine) palladium (20 mg,0.05 mmol), potassium carbonate (75 mg,0.54 mmol) were dissolved in dioxane (6 mL) and water (1 mL) and stirred under nitrogen atmosphere at 80℃for 2 hours. The reaction mixture was diluted with water (20 mL), extracted with ethyl acetate (50 mL), and the organic phase was washed with water (10 mL) and saturated brine (10 mL). Drying over anhydrous sodium sulfate, filtering, desolventizing under reduced pressure, and separating the residue by silica gel column chromatography (dichloromethane: methanol=100:0-9:1) to obtain target product ((3 s,4 s) -1- (5- (7-chloroquinolin-5-yl) pyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamic acid tert-butyl ester (0.11 g, yellow liquid), yield: 90 Percent of the total weight of the composition. MS M/z (ESI) 455 [ M+1 ];

Third step

(3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester

Tert-butyl (3 s,4 s) -1- (5- (7-chloroquinolin-5-yl) pyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamate (0.11 g,0.24 mmol), 1-methyl-1H-pyrazole-4-boronic acid pinacol ester (0.10 g,0.48 mmol), tetrakis (triphenylphosphine) palladium (20 mg,0.02 mmol), potassium phosphate (0.10 g,0.48 mmol) in dioxane (4 mL) and water (1 mL), oil bath 120 ℃ stirring overnight under nitrogen atmosphere conditions the reaction solution was diluted with water (20 mL), extracted with ethyl acetate (50 mL), the organic phase was washed with water (10 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, desolventized under reduced pressure, and the residue was chromatographed on silica gel (dichloromethane: methanol=100:0 to 20:1) to give the title product (3 s,4 s) -3-hydroxy-1- (5- (7-methyl-1H-4-pyridinyl) 2-yl) carbamate as a yellow solid (3912. 0.12 g-tert-butyl) carbamate: 99 The%;

MS m/z (ESI): 501 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 8.88 (d, J = 4.0 Hz, 1H), 8.28 (d, J = 2.0 Hz, 1H), 8.18-8.16 (m, 2H), 7.93 (s, 1H), 7.79 (s, 1H), 7.61 (d, J = 2.0 Hz, 1H), 7.60-7.58 (m, 1H), 7.31-7.29 (m, 1H), 6.84 (d, J = 8.8 Hz, 1H), 4.90-4.88 (m, 1H), 4.50-4.43 (m, 2H), 3.98 (s, 3H), 3.63-3.57 (m, 2H), 3.49-3.47 (m, 2H), 3.00-2.87 (m, 1H), 2.14-2.10 (m, 1H), 1.47 (s, 9H);

Fourth step

(3S, 4S) -4-amino-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-3-ol

The compound (3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester (0.11 g,0.22 mmol) was dissolved in dioxane (4 mL,16 mmol, 4M) of hydrochloric acid and stirred at room temperature for 30 minutes. The reaction mixture was desolventized under reduced pressure, neutralized with a saturated aqueous sodium hydrogencarbonate (3 mL), extracted with ethyl acetate (10 mL), and the organic phase was washed with saturated brine (4 mL ×3). Drying over anhydrous sodium sulfate, filtration, and desolventizing under reduced pressure to give the target product (3 s,4 s) -4-amino-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-3-ol (70 mg, yellow solid), yield: 80 Percent of the total weight of the composition. MS M/z (ESI) 401 [ M+1 ];

Fifth step

The compounds o-chlorobenzoic acid (4 mg,0.02 mmol), triethylamine (5 mg,0.05 mmol) and 2- (7-oxybenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (7 mg,0.02 mmol) were dissolved in N, N-dimethylformamide (1 mL mmol), and stirred at room temperature for 5 minutes. (3S, 4S) -4-amino-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-3-ol (5 mg,0.01 mmol) was added and stirring was continued at room temperature for 10 minutes. The reaction solution was washed with saturated sodium hydroxide solution (2 mL), extracted with ethyl acetate (10 mL), the organic phase was washed with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and desolventized under reduced pressure to give the target product 2-chloro-N- ((3 s,4 s) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-yl) benzamide (2.7 mg, white solid), yield: 42 The%;

MS m/z (ESI): 539&541 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 8.90-8.89 (m, 1H), 8.32 (d, J = 2.0 Hz, 1H), 8.18 (s, 1H), 7.93 (s, 1H), 7.80 (s, 1H), 7.77-7.75 (m, 1H), 7.66-7.64 (m, 1H), 7.60 (d, J = 1.6 Hz, 1H), 7.53 (d, J = 8.4 Hz, 1H), 7.45-7.35 (m, 3H), 7.33-7.30 (m, 1H), 6.89 (d, J = 8.8 Hz, 1H), 6.42 (d, J = 6.4 Hz, 1H), 4.20-4.15 (m, 1H), 4.00 (s, 3H), 3.73-2.99 (m, 5H), 2.05-2.00 (m, 2H).

Example 44

2-Chloro-N- (4- ((4-ethylpiperazin-1-yl) methyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-yl) -6-fluorobenzamide

First step

1- (5-Bromopyridin-2-yl) -4- ((tert-butoxycarbonyl) amino) piperidine-4-carboxylic acid methyl ester

The compound 2-fluoro-5-bromopyridine (0.61 g,3.49 mmol), methyl 4- ((tert-butoxycarbonyl) amino) piperidine-4-carboxylate (0.90 g,3.49 mmol), potassium carbonate (1.60 g,13.94 mmol) and dimethyl sulfoxide (15 mL) were mixed and stirred at 100℃for 16 hours. Cooled to room temperature, the mixture was slowly poured into water (40 mL), extracted with ethyl acetate (50 mL ×3), the organic phase was washed with water (50 mL ×3) and saturated brine (50 mL ×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure to give methyl 1- (5-bromopyridin-2-yl) -4- ((tert-butoxycarbonyl) amino) piperidine-4-carboxylate (1.10 g, crude). Directly used in the next step. MS M/z (ESI) 414&416 [ M+1 ];

Second step

(1- (5-Bromopyridin-2-yl) -4- (hydroxymethyl) piperidin-4-yl) carbamic acid tert-butyl ester

The compound methyl 1- (5-bromopyridin-2-yl) -4- ((tert-butoxycarbonyl) amino) piperidine-4-carboxylate (0.25 g,0.60 mmol) was dissolved in tetrahydrofuran (3 mL), a solution of lithium borohydride in tetrahydrofuran (2 mL, 2M) was added at room temperature, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (20 mL ×3), and the organic phase was washed with water (20 mL ×3) and saturated brine (20 mL ×3). Drying over anhydrous sodium sulfate, filtering to remove the drying agent, decompressing and desolventizing the filtrate, purifying the residue by a silica gel chromatographic column (petroleum ether: ethyl acetate=100:0-4:6) to obtain the target product (tert-butyl 1- (5-bromopyridin-2-yl) -4- (hydroxymethyl) piperidin-4-yl) carbamate (0.14 g, white solid), yield: 56 Percent of the total weight of the composition. MS M/z (ESI) 386&388 [ M+1 ];

Third step

(1- (5-Bromopyridin-2-yl) -4-formylpiperidin-4-yl) carbamic acid tert-butyl ester

The compound (1- (5-bromopyridin-2-yl) -4- (hydroxymethyl) piperidin-4-yl) carbamic acid tert-butyl ester (0.93 g,2.59 mmol) was dissolved in dichloromethane (10 mL), and dess-martin reagent (1.65 g,3.89 mmol) was added in portions and stirred at room temperature for 4 hours. The reaction solution was washed with a saturated sodium sulfite (5 mL) solution, extracted with dichloromethane (20 mL), and the organic phase was washed with water (20 mL) and saturated brine (20 mL). Drying over anhydrous sodium sulfate, filtering, and desolventizing under reduced pressure to obtain the target product (1- (5-bromopyridin-2-yl) -4-formylpiperidin-4-yl) carbamic acid tert-butyl ester (0.68 g, white solid), yield: 68 The%;

MS m/z (ESI): 384&386 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.52 (s, 1H), 8.18 (d, J = 2.0 Hz, 1H), 7.55-7.52 (m, 1H), 6.58 (d, J = 8.8 Hz, 1H), 4.92 (s, 1H), 3.94-3.88 (m, 2H), 3.36-3.33 (m, 2H), 2.04-1.98 (m, 2H), 1.87-1.84 (m, 2H), 1.45 (s, 9H);

Fourth step

(1- (5-Bromopyridin-2-yl) -4- ((4-ethylpiperazin-1-yl) methyl) piperidin-4-yl) carbamic acid tert-butyl ester

The compound (1- (5-bromopyridin-2-yl) -4-formylpiperidin-4-yl) carbamic acid tert-butyl ester (0.60 g,1.60 mmol) and 2-ethylpiperazine (0.34 g,3.00 mmol) were dissolved in tetrahydrofuran (15 mL), to which was added tetraethyltitanate (0.94 g,4.16 mmol), and after stirring at room temperature for 30 minutes, sodium borohydride acetate (0.85 g,4.00 mmol) was added and reacted at 50℃for 3 hours. After the reaction mixture was diluted with dichloromethane/methanol (10:1, 200 mL), water (5 mL) was added, and after stirring vigorously for 15 minutes, the mixture was filtered, the filter cake was washed with dichloromethane (40 mL ×3), and the organic phases were combined and washed with water (20 mL) and saturated brine (20 mL). Drying with anhydrous sodium sulfate, filtering to remove desiccant, and removing solvent under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane: methanol=20:1) to give tert-butyl carbamate (0.53 g, yellow oil) as the target product (1- (5-bromopyridin-2-yl) -4- ((4-ethylpiperazin-1-yl) methyl) piperidin-4-yl), yield: 70 Percent of the total weight of the composition. MS M/z (ESI): 482&484 [ M+1 ];

Fifth step

(1- (5- (7-Chloroquinolin-5-yl) piperidin-2-yl) -4- ((4-ethylpiperazin-1-yl) methyl) piperidin-4-yl) carbamic acid tert-butyl ester

7-Chloro-5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) quinoline (0.17 g,0.60 mmol), (1- (5-bromopyridin-2-yl) -4- ((4-ethylpiperazin-1-yl) methyl) piperidin-4-yl) carbamic acid tert-butyl ester (0.20 g,0.41 mmol) and dioxane (10 mL), water (2 mL) were mixed, potassium carbonate (0.17 g,1.23 mmol) and tetrakis (triphenylphosphine) palladium (47 mg,0.041 mmol) were added under argon atmosphere, replaced three times with argon atmosphere, and stirred at 80℃for 5 hours under argon atmosphere. Cooled to room temperature, desolventized under reduced pressure, the residue was taken up in water (20 mL), extracted with ethyl acetate (15 mL ×3), and the organic phase was washed with saturated brine (20 mL ×3). Drying over anhydrous sodium sulfate, filtering to remove the drying agent, decompressing and desolventizing to obtain crude product, purifying (dichloromethane: methanol=100:0-15:1) by flash silica gel column chromatography to obtain target product (tert-butyl 1- (5- (7-chloroquinolin-5-yl) piperidin-2-yl) -4- ((4-ethylpiperazin-1-yl) methyl) piperidin-4-yl) carbamate (0.10 g, yellow solid), yield: 43 Percent of the total weight of the composition. MS M/z (ESI): 565&567 [ M+1 ];

Sixth step

(4- ((4-Ethylpiperazin-1-yl) methyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester

The compound (1- (5- (7-chloroquinolin-5-yl) piperidin-2-yl) -4- ((4-ethylpiperazin-1-yl) methyl) piperidin-4-yl) carbamic acid tert-butyl ester (0.10 g,0.18 mmol), 1-methyl-1H-pyrazole-4-boronic acid pinacol ester (75 mg,0.36 mmol) and dioxane (6 mL) and water (1 mL) were mixed into a 35 mL sealed tube, potassium carbonate (75 mg,0.54 mmol) and tetrakis (triphenylphosphine) palladium (21 mg,0.018 mmol) were added under argon atmosphere, replaced three times with argon, and stirred for 3 hours at 140℃under argon atmosphere. Cooled to room temperature, desolventized under reduced pressure, the residue was taken up in water (20 mL), extracted with ethyl acetate (15 mL ×3), and the organic phase was washed with saturated brine (20 mL ×2). Drying over anhydrous sodium sulfate, filtering to remove the drying agent, and removing the solvent under reduced pressure to obtain crude product, which is purified by silica gel preparation plate (dichloromethane/methanol=12:1) to obtain target product (4- ((4-ethylpiperazin-1-yl) methyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester (50 mg, yellow solid), yield: 45 Percent of the total weight of the composition. MS M/z (ESI) 611 [ M+1 ];

Seventh step

4- ((4-Ethylpiperazin-1-yl) methyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-amine

The compound (4- ((4-ethylpiperazin-1-yl) methyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester (50 mg,0.082 mmol) and a solution of hydrogen chloride in methanol (6 mL,24 mmol, 4M) were mixed and stirred at room temperature for 1 hour. The residue was desolventized under reduced pressure, taken up in saturated sodium bicarbonate solution (10 mL), extracted with dichloromethane (10 mL ×4), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the desired product 4- ((4-ethylpiperazin-1-yl) methyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-amine (35 mg, yellow solid) was obtained by desolventizing under reduced pressure and was used directly in the next reaction without purification. MS M/z (ESI): 511 [ M+1 ];

eighth step

The compound 4- ((4-ethylpiperazin-1-yl) methyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-amine (6.0 mg,0.012 mmol), 2-chloro-6-fluorobenzoic acid (2.6 mg,0.015 mmol), 2- (7-oxobenzotriazol) -N, N' -tetramethylurea hexafluorophosphate (6.8 mg,0.018 mmol), triethylamine (3 mg,0.03 mmol) and N, N-dimethylformamide (2 mL) were mixed and stirred at room temperature for 1 hour. Water (20 mL) was added to dilute the mixture, the mixture was extracted with methylene chloride (20 mL X4), and the organic phase was washed with saturated brine (20 mL X2). Drying over anhydrous sodium sulfate, filtration to remove the drying agent, and purification of the residue from the preparative liquid phase (Agilent ZORBAX XDB-C18, 4.6x50 mm,3.5 μm, ACN/H ₂ O25% -50%) afforded the desired product 2-chloro-N- (4- ((4-ethylpiperazin-1-yl) methyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) quinolin-5-yl) pyridin-2-yl) piperidin-4-fluorobenzamide (2.2 mg, white solid), yield: 28 The%;

MS m/z (ESI): 667&669 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 8.92-8.90 (m, 1H), 8.32 (d, J = 2.4 Hz, 1H), 8.27 (s, 1H), 8.26-8.18 (m, 2H), 7.94 (s, 1H), 7.82 (s, 1H), 7.67-7.59 (m, 2H), 7.38-7.30 (m, 2H), 7.09-7.06 (m, 1H), 6.85 (d, J = 8.8 Hz, 1H), 6.08 (s, 1H), 4.34-4.18 (m, 2H), 4.00 (s, 3H), 3.40-3.27 (m, 2H), 3.17-3.01 (m, 10H), 2.98 (s, 2H), 2.54-2.41 (m, 2H), 1.82-1.69 (m, 2H), 1.40-1.31 (m, 3H).

Example 142

2-Chloro-N- ((3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) benzamide

First step

((3S, 4S) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamic acid tert-butyl ester

The compound 5, 7-dichloro-1, 6-naphthyridine (synthetic reference: PCT Int. Appl., 2011134971) (45 mg,0.23 mmol), (6- ((3S, 4S) -4-carbamic acid tert-butyl ester-3-hydroxypiperidin-1-yl) pyridin-3-yl) boronic acid (crude) and potassium carbonate (89 mg,0.65 mmol) were added to 1, 4-dioxane (5 mL) and water (1 mL), and tetrakis (triphenylphosphine) palladium (25 mg,0.022 mmol) was added under nitrogen protection and reacted at 90℃for 12 hours. The reaction solution was diluted with ethyl acetate (80 mL), and washed with water (10 mL ×3) and saturated brine (10 mL ×3). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the solvent was removed under reduced pressure. Purification of the residue on a preparative silica gel plate (petroleum ether/ethyl acetate 1:1) afforded the desired product ((3 s,4 s) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamic acid tert-butyl ester (60 mg, yellow solid) in 61% yield;

MS m/z (ESI): 456&458 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.07 (dd, J = 4.2, 1.6 Hz, 1H), 8.50-8.46 (m, 2H), 7.96-7.88 (m, 2H), 7.70-7.61 (m, 1H), 6.85 (d, J = 8.8 Hz, 1H), 4.78 (d, J = 6.4 Hz, 1H), 4.59-4.42 (m, 2H), 3.66-3.59 (m, 1H), 3.56-3.42 (m, 1H), 3.02-2.88 (m, 2H), 2.10-2.05 (m, 2H), 1.47 (s, 9H);

Second step

((3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester

The compound (((3S, 4S) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamic acid tert-butyl ester (25 mg,0.055 mmol), 1-methyl-1H-pyrazole-4-boronic acid pinacol ester (29 mg,0.137 mmol) and potassium phosphate (33 mg,0.165 mmol) were dissolved in dioxane (2 mL) and water (0.5 mL), palladium tetrakis (triphenylphosphine) was added under nitrogen protection (7 mg,0.006 mmol), the mixture was diluted with dichloromethane/methanol (20:1, 50 mL) at 140℃for 1.5 hours, the saturated brine (5 mL X3) and the organic phase was dried over anhydrous sodium sulfate, the desiccant was removed by filtration, and the residue was purified with a preparative silica gel plate (dichloromethane/methanol 25:1) to give the target product ((3S) -3-hydroxy-1- (7 mg,0.006 mmol) palladium (34%: 6-naphthyridin-1- (1H-5-naphthyridin-5-yl) 1-yl) amino-34% [ 34M-4-5-yl ] piperidine ] solid (34M-4-1, 34%) [ 1-naphthyridine);

Third step

(3S, 4S) -4-amino-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-3-ol

The compound ((3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester (15 mg,0.030 mmol) was dissolved in ethyl acetate (2 mL) and ethyl acetate hydrochloride solution (8 mL,16 mml, 2M) was added at room temperature. The reaction was stirred at room temperature for 0.5 hour. The reaction mixture was desolventized under reduced pressure, and the residue was dissolved in methylene chloride/methanol (10:1), and the pH was adjusted to about 8-9 with triethylamine. The mixture is rotary distilled to remove solvent, and then yellow solid crude product is obtained and directly used for the next reaction. MS M/z (ESI) 402 [ M+1 ];

Fourth step

The compound (3 s,4 s) -4-amino-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-3-ol (crude, 0.030 mmol), 2-chlorobenzoic acid (7 mg,0.045 mmol) and triethylamine (6 mg,0.06 mmol) were dissolved in dichloromethane (2 mL), 2- (7-benzotriazol-oxide) -N, N' -tetramethylurea hexafluorophosphate (17 mg,0.045 mmol) was added at room temperature and the reaction stirred at room temperature for 0.5 hours. The reaction mixture was diluted with water (20 mL), extracted with dichloromethane (20 mL ×2), and the combined organic phases were washed with saturated brine (20 mL ×2). Drying over anhydrous sodium sulfate, filtration, desolventizing under reduced pressure, and purification of the residue using preparative silica gel plate (dichloromethane/methanol 30:1) gives the title product 2-chloro-N- ((3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) benzamide (10 mg, yellow solid). Yield: 62 The%;

MS m/z (ESI): 540&542 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.02-9.00 (m, 1H), 8.57 (d, J = 3.2 Hz, 1H), 8.43 (d, J = 8.2 Hz, 1H), 8.12 (s, 1H), 8.08 (s, 1H), 7.98-7.95 (m, 2H), 7.75-7.73 (m, 1H), 7.42-7.35 (m, 3H), 6.91 (d, J = 8.8 Hz, 1H), 6.64 (d, J = 6.8 Hz, 1H), 4.65-4.53 (m, 2H), 4.27-4.25 (m, 1H), 4.17-4.15 (m, 1H), 3.99 (s, 3H), 3.77-3.71 (m, 1H), 3.30-3.24 (m, 1H), 2.96-2.90 (m, 1H), 2.32-2.27 (m, 1H).

synthetic procedure of examples 143 to 167 reference is made to the last step of example 142, in which 2-chlorobenzoic acid is replaced by a different acid:

。

Example 168

2-Chloro-N- ((3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyrazin-2-yl) piperidin-4-yl) benzamide

First step

(6- ((3S, 4S) -4- ((tert-Butoxycarbonyl) amino) -3-hydroxypiperidin-1-yl) pyrazin-3-yl) boronic acid

Tert-butyl (3S, 4S) -1- (5-bromopyrazin-2-yl) -3-hydroxypiperidin-4-yl) carbamate (first step of Synthesis reference example 23) (0.52 g,1.40 mmol), pinacol diboronate (0.71 g,2.80 mmol) was dissolved in 1, 4-dioxane (20 mL), tris (dibenzylideneacetone) dipalladium (0.14 g,0.15 mmol), tricyclohexylphosphine (84 mg,0.30 mmol) and potassium acetate (0.41 g,4.20 mmol) were added, the reaction solution was bubbled with nitrogen for ten minutes, and heated and stirred under nitrogen at 95℃for 2 hours. The reaction solution was spin-dried to give the crude (6- ((3 s,4 s) -4- ((tert-butoxycarbonyl) amino) -3-hydroxypiperidin-1-yl) pyrazin-3-yl) boronic acid (1.20 g, crude) which was used directly in the next step without purification. MS M/z (ESI) 339 [ M+1 ];

Second step

((3S, 4S) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyrazin-2-yl) -3-hydroxypiperidin-4-yl) carbamic acid tert-butyl ester

(6- ((3S, 4S) -4- ((tert-Butoxycarbonyl) amino) -3-hydroxypyrazin-1-yl) pyridin-3-yl) boronic acid (1.20 g, crude), 5, 7-dichloro-1, 6-naphthyridine (synthetic reference: PCT Int. Appl., 2011134971) (0.30 g,1.50 mmol), dissolved in 1, 4-dioxane (5 mL) and water (1 mL), and tris (dibenzylideneacetone) dipalladium (92 mg,0.10 mmol), tricyclohexylphosphine (56 mg,0.20 mmol) and potassium carbonate (0.62 g,4.50 mmol) were added, the reaction solution was bubbled with nitrogen for ten minutes and heated to 90℃under nitrogen for 14 hours. Cooled to room temperature, diluted with dichloromethane (100 mL), the organic phase washed with water (20 mL ×3) and saturated brine (20 mL ×3), dried over anhydrous sodium sulfate, filtered, desolventized under reduced pressure, and the crude product purified with a silica gel column (petroleum ether: ethyl acetate=1:1) to give ((3 s,4 s) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyrazin-2-yl) -3-hydroxypiperidin-4-yl) carbamic acid tert-butyl ester (0.28 g, yellow solid) in 43.7% yield;

MS m/z (ESI): 457&459 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.39 (d, J = 8.8 Hz, 1H), 9.06-9.04 (m, 1H), 9.00 (s, 1H), 8.26 (s, 1H), 8.19-8.16 (m, 1H), 7.94 (s, 1H), 4.76-4.74 (m, 1H), 4.58-4.52 (m, 2H), 3.55-3.48 (m, 1H), 3.06-2.80 (m, 2H), 2.12-2.07 (m, 1H), 1.57-1.51 (m, 1H), 1.47 (s, 9H);

Third step

((3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyrazin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester

Tert-butyl ((3S, 4S) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyrazin-2-yl) -3-hydroxypiperidin-4-yl) carbamate (0.28 g,0.62 mmol), 1-methyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (0.26 g,1.24 mmol) was dissolved in N, N-dimethylacetamide (5 mL) and water (1 mL), the reaction mixture was bubbled with nitrogen for ten minutes, and heated and stirred at 120℃for 4 hours under nitrogen. The reaction solution was cooled to room temperature, diluted with dichloromethane (100 mL), and the organic phase was washed with water (20 mL) and saturated brine (20 mL). Drying over anhydrous sodium sulfate, filtering, and desolventizing under reduced pressure, purifying the crude product obtained by column chromatography on silica gel (dichloromethane: methanol=40:1) to give tert-butyl ((3 s,4 s) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyrazin-2-yl) piperidin-4-yl) carbamate (100 mg, yellow solid) as the target product in 32.7% yield;

MS m/z (ESI): 503 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.28 (d, J = 8.8 Hz, 1H), 9.13 (s, 1H), 8.99-8.97 (m, 1H), 8.27 (s, 1H), 8.14 (s, 1H), 8.09 (s, 1H), 7.98 (s, 1H), 7.44-7.40 (m, 1H), 4.89-4.87 (m, 1H), 4.57-4.53 (m, 2H), 3.97 (s, 3H), 3.54-3.51 (m, 1H), 3.09-2.94 (m, 2H), 2.15-2.11 (m, 1H), 1.59-1.51 (m, 1H), 1.47 (s, 9H);

Fourth step

(3S, 4S) -4-amino-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyrazin-2-yl) piperidin-3-ol hydrochloride

Tert-butyl ((3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyrazin-2-yl) piperidin-4-yl) carbamate (0.10 g,0.20 mmol) was dissolved in dioxane hydrochloride solution (5 mL,20 mmol, 4M) and stirred at room temperature for 1 hour. The reaction solution was spin-dried to give the crude (3 s,4 s) -4-amino-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyrazin-2-yl) piperidin-3-ol hydrochloride (110 mg, crude) which was used directly in the next step without purification. MS M/z (ESI) 403 [ M+1 ];

Fifth step

The compound (3 s,4 s) -4-amino-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyrazin-2-yl) piperidin-3-ol hydrochloride (crude, 0.004 mmol), 2-chlorobenzoic acid (1 mg,0.006 mmol) and triethylamine (1 mg,0.008 mmol) were dissolved in dichloromethane (1 mL), 2- (7-benzotriazol-oxide) -N, N' -tetramethylurea hexafluorophosphate (2.3 mg,0.006 mmol) was added at room temperature and the reaction was stirred at room temperature for 0.5 hours. The reaction mixture was diluted with water (5 mL), dichloromethane (10 mL X2) was used for extraction, the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, desolventized under reduced pressure, and the residue was purified using preparative silica gel plates (dichloromethane/methanol 25:1) to give the title product 2-chloro-N- ((3S, 4S) -3-hydroxy-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyrazin-2-yl) piperidin-4-yl) benzamide (1.2 mg, yellow solid). Yield: 50 The%;

MS m/z (ESI): 541&543 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.31 (d, J = 7.8 Hz, 1H), 9.10 (d, J = 1.2 Hz, 1H), 9.01-9.00 (m, 1H), 8.32 (s, 1H), 8.14 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.79-7.73 (m, 1H), 7.45-7.36 (m, 3H), 6.45 (d, J = 6.8 Hz, 1H), 4.68-4.56 (m, 2H), 4.30-4.17 (m, 1H), 4.00 (s, 3H), 3.81-3.68 (m, 1H), 3.18-3.06 (m, 2H), 2.29-2.26 (m, 1H), 2.01-1.98 (m, 1H).

Synthetic procedure of examples 169 to 181 refer to the final step of example 27, in which 2-chlorobenzoic acid is replaced with a different acid:

。

Example 183

2-Chloro-N- (1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) benzamide

First step

(1- (5- (7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester

Tert-butyl (1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamate (third step of synthetic reference example 168) (38 mg,0.09 mmol), 1-methyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (37 mg,0.18 mmol) was dissolved in N, N-dimethylformamide (4 mL), tetrakis (triphenylphosphine) palladium (10 mg,0.009 mmol) and potassium phosphate (57 mg,0.27 mmol) were added in sequence, and the mixture was heated to 140℃under nitrogen protection to react for 6 hours. The reaction solution was cooled to room temperature, poured into water (20 mL), extracted with ethyl acetate (25 mL ×3), the organic phases were combined, washed with saturated brine (25 mL ×3), dried over anhydrous sodium sulfate, filtered and concentrated, and purified by thin layer chromatography (dichloromethane: methanol=15:1) to give tert-butyl (37 mg, white solid) carbamate (1- (5- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) of the target compound in 84.7% yield;

MS m/z (ESI): 486 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.00 (s, 1H), 8.57 (m, 1H), 8.44-8.42 (m, 1H), 8.12 (s, 1H), 8.07 (s, 1H), 7.96-7.95 (m, 2H), 7.38-7.35 (m, 1H), 6.83 (d, J = 8.8 Hz, 1H), 4.53-4.52 (m, 1H), 4.37-4.35 (m, 2H), 3.99 (s, 3H), 3.77-3.76 (m, 1H), 3.14-3.08 (m, 2H), 2.10-2.05 (m, 4H), 1.47 (s, 9H);

Second step

(1- (5- (7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) amino hydrochloride

(1- (5- (7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester (10 mg,0.02 mmol) was dissolved in methanol (3 mL), dioxane hydrochloride solution (3 mL,12 mmol, 4M) was added, and the mixture was reacted at room temperature for 30 minutes. The reaction solution was concentrated to give crude (1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) amino hydrochloride (crude, 10 mg, yellow solid) of the target compound, which was directly fed to the next step. MS M/z (ESI) 386 [ M+1 ];

Third step

O-chlorobenzoic acid (6 mg,0.04 mmol), 2- (7-benzotriazol-N, N, N ', N' -tetramethylurea hexafluorophosphate (16 mg,0.04 mmol), triethylamine (0.1 mL) were dissolved in dichloromethane (5 mL), and (1- (5- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) amino hydrochloride (10 mg,0.02 mmol) was added and the reaction was stirred at room temperature for 1 hour. Dichloromethane (20 mL) was added for dilution, the organic phase was washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered and concentrated, and purified by thin layer chromatography (dichloromethane: methanol=15:1) to give 2-chloro-N- (1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) benzamide (8.0 mg, yellow solid) in 76.5% yield;

MS m/z (ESI): 524&526 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.01-9.00 (m, 1H), 8.59-8.58 (m, 1H), 8.43 (d, J = 8.4 Hz, 1H), 8.12 (s, 1H), 8.07 (s, 1H), 7.98-7.95 (m, 2H), 7.69-7.66 (m, 1H), 7.41-7.32 (m, 4H), 6.87 (d, J = 8.8 Hz, 1H), 6.22 (d, J = 8.0 Hz, 1H), 4.43-4.34 (m, 3H), 3.99 (s, 3H), 3.27-3.21 (m, 2H), 2.23-2.21 (m, 2H), 1.67-1.62 (m, 2H).

Synthetic procedure of examples 184 to 203 refer to the last step of example 183, in which 2-chlorobenzoic acid was replaced with a different acid:

。

Example 205

2-Chloro-N- ((3S, 4S) -3-hydroxy-1- (5- (7- ((1-methylpiperidin-4-yl) methoxy) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) benzamide

First step

((3S, 4S) -3-hydroxy-1- (5- (7- ((1-methylpiperidin-4-yl) methoxy) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester

Tert-butyl ((3S, 4S) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamate (46 mg,0.10 mmol), 1-methyl-4-piperidinemethanol (65 mg,0.05 mmol) was dissolved in 1, 4-dioxane (1.5 mL), tris (dibenzylideneacetone) dipalladium (9.2 mg,0.01 mmol) and cesium carbonate (98 mg,0.30 mmol) were added, the reaction mixture was bubbled with nitrogen for ten minutes and stirred under microwave heating at 110℃under nitrogen for 1 hour. Dichloromethane (20 mL) was added thereto, and the mixture was washed with saturated brine (20 mL ×2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was desolventized under reduced pressure. Purification of the residue on a silica gel column (dichloromethane: methanol=15:1) afforded tert-butyl ((3 s,4 s) -3-hydroxy-1- (5- (7- ((1-methylpiperidin-4-yl) methoxy) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamate (8 mg, yellow solid) as the target product in 20.2% yield;

MS m/z (ESI): 549 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 8.95 (d, J = 2.8 Hz, 1H), 8.50 (d, J = 2.0 Hz, 1H), 8.41 (d, J = 8.8 Hz, 1H), 8.34 (s, 1H), 7.91-7.88 (m, 1H), 7.30-7.27 (m, 1H), 7.20 (s, 1H), 6.86 (d, J = 8.8 Hz, 1H), 4.78-4.48 (m, 3H), 4.33 (d, J = 6.0 Hz, 2H), 3.66-3.51 (m, 4H), 3.02-2.89 (m, 2H), 2.76-2.70 (m, 5H), 2.13-2.04 (m, 4H), 1.92-1.84 (m, 2H), 1.47 (s, 9H);

Second step

(3S, 4S) -4-amino-1- (5- (7- ((1-methylpiperidin-4-yl) methoxy) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-3-ol

Tert-butyl ((3S, 4S) -3-hydroxy-1- (5- (7- ((1-methylpiperidin-4-yl) methoxy) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamate (8 mg,0.02 mmol) was dissolved in dioxane hydrochloride solution (1 mL,4 mmol, 4M) and stirred at room temperature for 1 hour. The reaction solution was neutralized by adding triethylamine and spin-dried to give the crude (3 s,4 s) -4-amino-1- (5- (7- ((1-methylpiperidin-4-yl) methoxy) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-3-ol (10 mg, crude) which was used directly in the next step without purification. MS M/z (ESI): 449 [ M+1 ];

Third step

The third operating step of reference example 183 is synthesized to yield the target product 2-chloro-N- ((3 s,4 s) -3-hydroxy-1- (5- (7- ((1-methylpiperidin-4-yl) methoxy) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) benzamide;

MS m/z (ESI): 587&589 [M + 1]；

¹H NMR (400 MHz, CD₃OD) δ 8.95-8.93 (m, 1H), 8.53-8.49 (m, 2H), 7.97-7.94 (m, 1H), 7.46-7.36 (m, 5H), 7.16 (s, 1H), 7.04 (d, J = 8.8 Hz, 1H), 4.65-4.58 (m, 1H), 4.42-4.39 (m, 3H), 4.11-4.05 (m, 1H), 3.67-3.62 (m, 1H), 3.54-3.48 (m, 2H), 3.21-3.13 (m, 1H), 3.07-2.97 (m, 2H), 2.86 (s, 3H), 2.17-2.10 (m, 4H), 1.75-1.61 (m, 4H).

synthetic procedure of examples 206 to 211 reference is made to the last step of example 205, in which 2-chlorobenzoic acid is replaced by a different acid:

。

Example 212

2-Chloro-N- (1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) benzamide

First step

(1- (5- (7-Ethoxy-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid ethyl ester

To absolute ethanol (5 mL) was added sodium metal (0.23 g,10 mmol), stirred at room temperature for 20 minutes, tert-butyl (1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamate (20 mg,0.046 mmol) was added, the reaction was continued for 2 hours at 80℃with a large amount of remaining starting material, the temperature was continued to increase to 100℃and the reaction was continued for 16 hours, and the crude (1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamate reaction solution was fed directly to the next step. MS M/z (ESI) 408 [ M+1 ];

Second step

(1- (5- (7-Ethoxy-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) amine

To a solution of crude ethyl (1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamate (crude, 0.046 mmol) ethanol (5 mL) was added water (3 mL), sodium hydroxide (5 mg,0.09 mmol) and the temperature was raised to 120℃for reaction for 5 hours. The reaction solution was dried by spinning, ethyl acetate (50 mL) was added, washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give (1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) amine (crude, 20 mg) which was directly fed to the next step. MS M/z (ESI) 350 [ M+1 ];

Third step

O-chlorobenzoic acid (18 mg,0.12 mmol), 2- (7-benzotriazol-N, N, N ', N' -tetramethylurea hexafluorophosphate (34 mg,0.09 mmol), triethylamine (10 mg,0.10 mmol) were dissolved in dichloromethane (5 mL), and (1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) amine (crude, 20mg, 0.06 mmol) was added and the reaction was stirred at room temperature for 1 hour. Dichloromethane (20 mL) was added for dilution, the organic phase was washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was prepared for liquid phase purification (Agilent ZORBAX XDB-C18, 4.6X50 mm,3.5 μm, ACN/H ₂ O25% -50%) to give the target product 2-chloro-N- (1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) benzamide (2.5 mg, yellow solid) in a total yield of 8.5% in three steps;

MS m/z (ESI): 488&490 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 8.96 (s, 1H), 8.54 (d, J = 2.0 Hz, 1H), 8.44 (d, J = 8.4 Hz, 1H), 7.97-7.94 (m, 1H), 7.69-7.67 (m, 1H), 7.40-7.33 (m, 4H), 7.25-7.21 (m, 1H), 6.86 (d, J = 9.2 Hz, 1H), 6.16 (d, J = 7.6 Hz, 1H), 4.49-4.39 (m, 5H), 3.27-3.21 (m, 2H), 2.24-2.22 (m, 2H), 1.65-1.62 (m, 2H), 1.27-1.23 (m, 3H).

Example 213

2-Chloro-N- ((3S, 4S) -1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl) pyridin-2-yl) -3-hydroxypiperidin-4-yl) benzamide

First step

((3S, 4S) -1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl)) pyridin-2-yl) -3-hydroxypiperidin-4-yl) amine

To absolute ethanol (5 mL) was added sodium metal (0.23 g,10 mmol), and the mixture was stirred at room temperature for 20 minutes, tert-butyl ((3S, 4S) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl)) pyridin-2-yl) -3-hydroxypiperidin-4-yl) carbamate (first step of synthesis reference example 142) (20 mg,0.046 mmol) was added and the mixture was allowed to react at 90℃for 16 hours. Concentrated under reduced pressure, ethyl acetate (20 mL) and water (20 mL) were added to the residue, extracted with ethyl acetate (20 mL ×2), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and desolventized under reduced pressure to give ((3 s,4 s) -1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl)) pyridin-2-yl) -3-hydroxypiperidin-4-yl) amine (crude, 20 mg), which was directly fed to the next step. MS M/z (ESI) 366 [ M+1 ];

Second step

O-chlorobenzoic acid (10 mg,0.055 mmol), 2- (7-oxybenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (42 mg,0.11 mmol), triethylamine (0.1 mL) were dissolved in dichloromethane (3 mL), and ((3S, 4S) -1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl)) pyridin-2-yl) -3-hydroxypiperidin-4-yl) amine (crude, 20 mg) was added and reacted at room temperature with stirring for 30 minutes. Dichloromethane (20 mL) was added thereto for dilution, and the mixture was washed with an aqueous lithium hydroxide solution (10 mL, 1M) and saturated brine (20 mL X2). Drying over anhydrous sodium sulfate, filtration, desolventizing under reduced pressure, and liquid phase purification of the residue preparation (Agilent ZORBAX XDB-C18, 4.6X10 mm,3.5 μm, ACN/H ₂ O20% -50%) gives the target product 2-chloro-N- ((3S, 4S) -1- (5- (7-ethoxy-1, 6-naphthyridin-5-yl) pyridin-2-yl) -3-hydroxypiperidin-4-yl) benzamide (2.0 mg, yellow solid), overall yield 9.9% in two steps;

MS m/z (ESI): 504&506 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 8.95-8.94 (m, 1H), 8.54 (d, J = 2.0 Hz, 1H), 8.39 (d, J = 8.8 Hz, 1H), 7.96-7.93 (m, 1H), 7.77-7.75 (m, 1H), 7.43-7.35 (m, 3H), 7.25-7.23 (m, 1H), 6.88 (d, J = 9.2 Hz, 1H), 6.40 (d, J = 6.4 Hz, 1H), 4.64-4.61 (m, 1H), 4.56-4.50 (m, 1H), 4.49-4.43 (m, 2H), 4.19-4.15 (m, 1H), 4.06-4.04 (m, 1H), 3.73-3.66 (m, 1H), 3.10-2.99 (m, 2H), 2.23-2.21 (m, 1H), 1.72-1.65 (m, 1H), 1.50-1.45 (m, 3H).

Example 214

5- (6- (4- (3-Fluorobenzyl) piperazin-1-yl) pyridin-3-yl) -7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridine formate salt

First step

4- (5- (7-Chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -1-piperazine-1-carboxylic acid tert-butyl ester

Tert-butyl (1- (5- (4, 5-tetramethyl-4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridin-2-yl) -1-piperazin-1-yl) carboxylate (0.17 g,0.45 mmol), 5, 7-dichloro-1, 6-naphthyridine (60 mg,0.3 mmol) was dissolved in dioxane (6 mL), tetrakis (triphenylphosphine) palladium (35 mg,0.03 mmol) and potassium carbonate (0.12 g,0.90 mmol) were added in sequence and heated to 85℃under nitrogen protection for 16 hours. The reaction solution was cooled to room temperature, filtered, concentrated, diluted with dichloromethane (50 mL), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, desolventized under reduced pressure, and purified (petroleum ether: ethyl acetate=1:1.5) to give the target compound tert-butyl 4- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -1-piperazine-1-carboxylate (110 mg, yellow solid) in 86.0% yield. MS M/z (ESI): 427&429 [ M+1 ];

¹H NMR (400 MHz, CDCl₃) δ 9.08-9.06 (m, 1H), 8.51-8.48 (m, 2H), 7.98-7.95 (m, 1H), 7.92 (s, 1H), 7.48-7.45 (m, 1H), 6.82 (d, J = 8.4 Hz, 1H), 3.70-3.68 (m, 4H), 3.60-3.56 (m, 4H), 1.49 (s, 9H);

Second step

4- (5- (7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -1-piperazine-1-carboxylic acid tert-butyl ester

Tert-butyl 4- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -1-piperazine-1-carboxylate (0.11 g,0.26 mmol), 1-methyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (0.11 g,0.52 mmol) was dissolved in N, N-dimethylacetamide (4 mL), tetrakis (triphenylphosphine) palladium (30 mg,0.026 mmol) and potassium phosphate (0.17 g,0.78 mmol) were added in sequence and heated to 140℃under nitrogen protection for reaction for 6 hours. The reaction solution was cooled to room temperature, poured into water (40 mL), extracted with ethyl acetate (50 mL ×3), the organic phases combined, washed with saturated brine (50 mL ×3), dried over anhydrous sodium sulfate, filtered, desolventized under reduced pressure, and purified on a silica gel preparation (dichloromethane: methanol=15:1) to give the target compound tert-butyl 4- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -1-piperazine-1-carboxylate (0.12 g, yellow solid) in 97.8% yield;

MS m/z (ESI): 472 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.02-9.01 (m, 1H), 8.59 (d, J = 2.4 Hz, 1H), 8.43-8.41 (m, 1H), 8.12 (s, 1H), 8.07 (s, 1H), 8.00-7.96 (m, 2H), 7.39-7.36 (m, 1H), 6.82 (d, J = 8.8 Hz, 1H), 3.98 (s, 3H), 3.91-3.68 (m, 4H), 3.61-3.59 (m, 4H), 1.51 (s, 9H);

Third step

4- (5- (7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -1-piperazine-1-carboxylic acid tert-butyl ester (0.12 g,0.25 mmol) was dissolved in methanol (5 mL), and dioxane hydrochloride solution (5 mL,20 mmol, 4M) was added and reacted at room temperature for 1.5 hours. The reaction solution was concentrated to obtain crude (1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) amino hydrochloride (crude, yellow solid) of the target compound, which was directly fed to the next step. MS M/z (ESI) 372 [ M+1 ];

Fourth step

(1- (5- (7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) amino hydrochloride (crude, 0.07 mmol) was dissolved in methanol (10 mL), triethylamine (10 mg,0.10 mmol) was added, stirred for 10 minutes, concentrated and then dissolved in methanol (10 mL), m-fluorobenzaldehyde (16 mg, 0.11 mmol) and sodium borohydride acetate (47 mg,0.21 mmol) were added in this order, and the mixture was reacted at room temperature for 48 hours. After the reaction solution is concentrated, dichloromethane (25 mL) is added for dissolution, saturated saline solution is used for washing (20 mL), anhydrous sodium sulfate is used for drying, filtration and decompression desolventizing are carried out, and a residue is prepared for liquid phase purification (Agilent ZORBAX XDB-C18, 4.6X50 mm,3.5 mu m, ACN/H ₂ O25% -50%) to obtain a target product 5- (6- (4- (3-fluorobenzyl) piperazin-1-yl) pyridin-3-yl) -7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridine formate (15 mg, yellow solid) with the yield of 50.6%;

MS m/z (ESI): 480 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.02-9.01 (m, 1H), 8.58 (d, J = 2.0 Hz, 1H), 8.45-8.43 (m, 1H), 8.27 (s, 1H), 8.13 (s, 1H), 8.07 (s, 1H), 7.98-7.96 (m, 2H), 7.41-7.38 (m, 1H), 7.36-7.30 (m, 1H), 7.18-7.13 (m, 2H), 7.04-7.02 (m, 1H), 6.82 (d, J = 8.8 Hz, 1H), 3.99 (s, 3H), 3.79-3.77 (m, 4H), 3.72 (m, 2H), 2.74 (t, J = 4.8 Hz, 4H).

synthesis of examples 215 to 222 reference the synthesis procedure of example 214:

。

example 223

2- (5-Fluoropyridin-2-yl) -1- (4- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperazin-1-yl) ethan-1-one

The compound 7- (1-methyl-1H-pyrazol-4-yl) -5- (6- (piperazin-1-yl) pyridin-3-yl) -1, 6-naphthyridine hydrochloride (20 mg,0.05 mmol) and triethylamine (10 mg,0.10 mmol) were dissolved in dichloromethane (2 mL), and 2- (5-fluoropyridin-2-yl) acetic acid (8 mg,0.05 mmol), 2- (7-oxobenzotriazol) -N, N' -tetramethylurea hexafluorophosphate (23 mg,0.06 mmol) was added, quenched with 10 mL water, the organic phase was separated, the aqueous phase extracted with dichloromethane (10 mL ×2) and the combined organic phases washed with saturated brine (20 mL ×2). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the solvent was removed under reduced pressure. Purification of the residue from the preparative liquid phase (Agilent ZORBAX XDB-C18, 4.6X150 mm,3.5 μm, ACN/H ₂ O ((0.1% TFA) 25% -55%) afforded the desired product 2- (5-fluoropyridin-2-yl) -1- (4- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperazin-1-one (5 mg, yellow solid): 19% yield;

MS m/z (ESI)：509 [M + 1];

¹H NMR (400 MHz, DMSO-d₆) δ 9.06-9.05 (m, 1H), 8.56-8.54 (m, 1H), 8.50-8.49 (m, 1H), 8.45 (s, 1H), 8.43-8.41 (m, 1H), 8.18 (s, 1H), 8.06(s, 1H), 8.03-8.00 (m, 1H), 7.72-7.67 (m, 1H), 7.56-7.53 (m, 1H), 7.43-7.40 (m, 1H), 7.06 (d, J = 8.8 Hz, 1H), 3.99 (s, 2H), 3.92 (s, 3H), 3.75-3.58 (m, 8H).

Synthesis procedure for examples 224 to 255 reference example 223 for synthesis: wherein 2- (5-fluoropyridin-2-yl) acetic acid is substituted with a different acid:

。

Example 256

5- (6- (4- (Ethylsulfonyl) piperazin-1-yl) pyridin-3-yl) -7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridine

The compound 7- (1-methyl-1H-pyrazol-4-yl) -5- (6- (piperazin-1-yl) pyridin-3-yl) -1, 6-naphthyridine hydrochloride (third step of synthesis reference example 214) (20 mg,0.054 mmol) was dissolved in N, N-dimethylformamide (2 mL), triethylamine (16 mg,0.162 mmol) and ethylsulfonyl chloride (9 mg,0.064 mmol) were added in this order, and after the addition, magnetic stirring was carried out at 25℃for 15 minutes. Quenched with water (10 mL), the mixture extracted with dichloromethane (10 mL ×2) and the combined organic phases washed with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the solvent was removed under reduced pressure. Purification of the residue by preparative high performance liquid chromatography (Agilent ZORBAX XDB-C18, 4.6X50 mm,3.5 μm, ACN/H ₂O（0.5 % NH₄ OH) 15% -35%) afforded the target product 5- (6- (4- (ethylsulfonyl) piperazin-1-yl) pyridin-3-yl) -7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridine (11.4 mg, yellow solid), yield: 46 The%;

MS m/z (ESI): 464 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.03-8.99 (m, 1H), 8.60 (d, J = 2.1 Hz, 1H), 8.42 (d, J = 8.0 Hz, 1H), 8.13 (s, 1H), 8.07 (s, 1H), 8.00-7.90 (m, 2H), 7.45-7.40 (m, 1H), 6.85 (d, J = 8.7 Hz, 1H), 3.99 (s, 3H), 3.85-3.74 (m, 4H), 3.50-3.41 (m, 4H), 3.02-2.95 (m, 2H), 1.47-1.40 (m, 3H).

Example 257

N-benzyl-4- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperazine-1-carboxamide

Benzylamine (11 mg,0.10 mmol) and triethylamine (25 mg,0.25 mmol) were dissolved in dichloromethane (5 mL), phenyl chloroformate (16 mg,0.10 mmol) was added and stirred at room temperature for 10 minutes, followed by 7- (1-methyl-1H-pyrazol-4-yl) -5- (6- (piperazin-1-yl) pyridin-3-yl) -1, 6-naphthyridine hydrochloride (third step of synthetic reference example 214) (20 mg,0.05 mmol). The temperature was raised to 60℃and the reaction was carried out for 5 hours. The residue was dissolved in ethyl acetate (20 mL), washed with saturated brine (20 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by preparative liquid phase (Agilent ZORBAX XDB-C18, 4.6x50 mm,3.5 μm, ACN/H ₂O（0.5 % NH₄ OH) 15% -35%) to give the desired product N-benzyl-4- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperazine-1-carboxamide (7 mg, yellow solid), yield: 28.2 The%;

MS m/z (ESI)：505 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.04-8.97 (m, 1H), 8.57 (s, 1H), 8.42 (d, J= 8.0 Hz, 1H), 8.11 (d, J = 12.0 Hz, 2H), 8.01-7.95 (m, 2H), 7.44-7.25 (m, 6H), 6.83 (d, J = 8.0 Hz, 1H), 4.46 (s, 2H), 3.99 (s, 3H), 3.80-3.72 (m, 4H), 3.63-3.59 (m, 4H).

synthesis of example 258 reference the procedure of example 257:

。

The synthesis procedure of examples 259 to 283 refers to the synthesis of example 223.

Example 284

3- (5- (7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 6-diazabicyclo [3.1.1] heptan-6-yl) (piperidin-4-yl) methanone hydrochloride

First step

4- (3- (5- (7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 6-diazabicyclo [3.1.1] heptane-6-carbonyl) piperidine-1-carboxylic acid tert-butyl ester

The compound 1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-amine (15 mg,0.04 mmol) was dissolved in dichloromethane (2 mL), and 1- (tert-butoxycarbonyl) piperidine-4-carboxylic acid (12 mg,0.05 mmol), 2- (7-benzotriazol) -N, N' -tetramethylurea hexafluorophosphate (19 mg,0.05 mmol), N-diisopropylethylamine (11 mg,0.08 mmol) was added and stirred at room temperature for 0.5 hours. Dichloromethane (20 mL) was added thereto, and the mixture was washed with water (20 mL ×2) and saturated brine (20 mL ×2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified from the preparative liquid phase (Agilent ZORBAX XDB-C18, 4.6x50 mm,3.5 μm, CH ₃CN/H₂ O ((0.1% TFA) 15% -30%) to give the desired product, tert-butyl 4- (3- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 6-diazabicyclo [3.1.1] heptane-6-carbonyl) piperidine-1-carboxylate (8.6 mg, yellow solid): yield: 36%. MS M/z (ESI): 595 [ m+1 ];

Second step

The compound 4- (3- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 6-diazabicyclo [3.1.1] heptane-6-carbonyl) piperidine-1-carboxylic acid tert-butyl ester (8.6 mg,0.01 mmol) was dissolved in 1, 4-dioxane solution (2 mL,8.00 mmol,4M) of hydrochloric acid and reacted at room temperature for 1 hour. The reaction mixture was concentrated directly to give the objective 3- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 6-diazabicyclo [3.1.1] heptan-6-yl) (piperidin-4-yl) methanone hydrochloride (5.4 mg, yellow solid). Yield: 78 The%;

MS m/z (ESI)：495 [M + 1];

¹H NMR (400 MHz, CD₃OD) δ 9.38-9.37 (m, 1H), 9.33-9.29 (m, 1H), 8.65-8.52 (m, 3H), 8.31 (s, 1H), 8.21 (s, 1H), 8.12-8.08 (m, 1H), 7.62-7.56 (m, 1H), 4.74-4.68 (m, 1H), 4.27-4.22 (m, 1H), 4.03 (s, 3H), 3.46-3.44 (m, 2H), 3.10-3.04 (m, 4H), 2.71-2.67 (m, 1H), 2.17-2.14 (m, 4H), 1.93-1.89 (m, 4H);

synthesis of example 285 reference the procedure of example 257:

。

Example 286

6- (3-Chlorobenzomethyl) -3- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 6-diazabicyclo [3.1.1] heptane

The compound 3- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 6-diazabicyclo [3.1.1] heptane (crude, 0.17 mmol) was dissolved in 1, 2-dichloroethane (2 mL), and 3-chlorobenzaldehyde (29 mg,0.20 mmol) and sodium borohydride acetate (72 mg,0.34 mmol) were added to react for 8 hours at ordinary temperature. The reaction was quenched by addition of saturated ammonium chloride solution, the organic phase was separated and the aqueous phase was extracted with dichloromethane (10 mL x 2). The combined organic phases were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative liquid phase (Agilent ZORBAX XDB-C18, 4.6x50 mm,3.5 μm, CH ₃CN/H₂ O ((0.1% TFA) 25% -55%) to give the desired product 6- (3-chlorophenyl) -3- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 6-diazabicyclo [3.1.1] heptane (3 mg, yellow solid): 3% yield;

MS m/z (ESI)：508&510 [M + 1];

¹H NMR (400 MHz, CD₃OD) δ 8.93-8.92 (m, 1H), 8.48-8.47 (m, 1H), 8.24 (s, 1H), 8.09 (s, 1H), 8.01-7.98 (m, 1H), 7.91 (s, 1H), 7.49-7.46 (m, 1H), 7.36 (s, 1H), 7.30-7.19 (m, 3H), 7.05-7.02 (m, 1H), 6.87 (d, J = 8.8 Hz, 1H), 4.03-3.98 (m, 1H), 3.90 (s, 3H), 3.87-3.84 (m, 2H), 3.72-3.66 (m, 3H), 2.11-2.07 (m, 1H), 1.71-1.69 (m, 1H), 1.52-1.49 (m, 2H).

Synthesis of example 287 reference the procedure of example 286:

。

Example 288

2- (5-Fluoropyridin-2-yl) -1- (3- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octan-8-yl) ethan-1-one

First step

3- (5-Bromopyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester

The compound 5-bromo-2-fluoropyridine (4.9 g,28.2 mmol), tert-butyl 3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (2.00 g,9.40 mmol) and N, N-diisopropylethylamine (3.60 g,28.20 mmol) were added to dimethyl sulfoxide (5 mL), and reacted overnight at 100 ℃. Cooled to room temperature, diluted with ethyl acetate (50 mL), washed with water (50 mL ×3) and saturated brine (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column separation (10-20% ethyl acetate/petroleum ether) to give the target product 3- (5-bromopyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester (3.10 g, white solid), yield: 89.5 The%;

MS m/z (ESI): 368&370 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 8.18 (d, J = 4.0 Hz, 1H), 7.52 (dd, J = 8.0, 4.0 Hz, 1H), 6.49 (d, J = 8.0 Hz, 1H), 4.46-4.52 (m, 2H), 3.88-3.58 (m, 2H), 3.26-3.03 (m, 2H), 2.041.76 (m, 4H), 1.45 (s, 9H);

Second step

3- (5- (7-Chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester

The compound 3- (5-bromopyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester (3.10 g,8.40 mmol), pinacol ester (3.20 g,12.60 mmol), potassium acetate (1.70 g,16.80 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (0.61 g,0.80 mmol) was added to dioxane (30 mL), the mixture was purged three times with nitrogen under vacuum, and the temperature was raised to 90℃for 4 hours. After cooling to room temperature, 5, 7-dichloro-1, 6-naphthyridine (1.70 g,8.40 mmol), tetrakis (triphenylphosphine) palladium (0.97 g,0.80 mmol), anhydrous potassium carbonate (2.30 g,16.60 mmol) and water (6 mL) were added and reacted overnight under nitrogen at 85 ℃. The residue was concentrated under reduced pressure, and then, the residue was dissolved in ethyl acetate (100 mL) and washed with water (100 mL) and saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (20-70% ethyl acetate/petroleum ether) to give the target product 3- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester (1.90 g, yellow solid). Yield: 49.8 The%;

MS m/z (ESI): 452&454 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.08 (d, J = 4.0 Hz, 1H), 8.56-8.46 (m, 2H), 7.93 (s, 1H), 7.72- 7.63 (m, 1H), 7.59-7.52 (m, 1H), 6.84-6.76 (m, 1H) 4.45-4.42 (m, 2H), 4.23- 3.96 (m, 2H), 3.27 (s, 2H), 2.00-1.98 (m, 2H), 1.85-1.75 (m, 2H), 1.50 (s, 9H);

Third step

3- (5- (7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester

A mixed solution of 3- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester (1.90 g,4.20 mmol), 1-methyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (1.70 g,8.40 mmol), potassium phosphate (1.80 g,8.40 mmol) and tetrakis triphenylphosphine palladium (0.49 g,0.40 mmol) was added to the mixture, and the mixture was purged three times with nitrogen under vacuum, and the mixture was reacted at 140℃for 4 hours. After cooling to room temperature, the reaction solution was diluted with ethyl acetate (100 mL), and then washed with saturated brine (80 mL ×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (20-100% ethyl acetate/petroleum ether) to give the target product 3- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester (1.40 g, yellow solid). Yield: 66.8 The%;

MS m/z (ESI): 498 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.05-9.00 (m, 1H), 8.62-8.58 (m, 1H), 8.12 (s, 1H), 8.08 (s, 1H), 8.00 (s, 1H), 7.72-7.63 (m, 1H), 7.59-7.52 (m, 2H), 6.84-6.76 (m, 1H), 4.44 (s, 2H), 4.28-4.06 (m, 2H), 3.99 (s, 3H), 3.27 (s, 2H), 2.04-1.98 (m, 2H), 1.86-1.78 (m, 2H), 1.50 (s, 9H);

Fourth step

5- (6- (3, 8-Diazabicyclo [3.2.1] oct-3-yl) pyridin-3-yl) -7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridine hydrochloride

The compound 3- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylic acid tert-butyl ester (1.40 g,2.80 mmol) was dissolved in a 1, 4-dioxane solution of hydrogen chloride (5 mL,20.00 mmol,4M), and stirred at room temperature for 1 hour. Concentrating under reduced pressure to obtain 5- (6- (3, 8-diazabicyclo [3.2.1] oct-3-yl) pyridin-3-yl) -7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridine hydrochloride (1.20 g, yellow solid), crude product. MS M/z (ESI): 398 [ M+1 ];

Fifth step

The compound 5- (6- (3, 8-diazabicyclo [3.2.1] oct-3-yl) pyridin-3-yl) -7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridine hydrochloride (17 mg,0.04 mmol) was dissolved in dichloromethane (2 mL mmol), and 2- (5-fluoropyridin-2-yl) acetic acid (8 mg,0.05 mmol), 2- (7-benzotriazol-oxide) -N, N, N ', N' -tetramethylurea hexafluorophosphate (19 mg,0.05 mmol), N, N-diisopropylethylamine (11 mg,0.08 mmol) was added and stirred at room temperature for 0.5 hours. Dichloromethane (20 mL) was added to dilute, saturated brine (20 mL ×2) was washed, and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Purification of the residue from the preparative liquid phase (Agilent ZORBAX XDB-C18, 4.6x50 mm,3.5 μm, CH ₃ CN/H2O ((0.1% TFA) 20% -40%) gave the desired product 2- (5-fluoropyridin-2-yl) -1- (3- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -3, 8-diazabicyclo [3.2.1] octan-8-yl) ethan-1-one (6 mg, yellow solid): 28% yield;

MS m/z (ESI)：535 [M + 1];

¹H NMR (400 MHz, CDCl₃) δ 9.02-9.01 (m, 1H), 8.58-8.57 (m, 1H), 8.42-8.40 (m, 2H), 8.12 (s, 1H), 8.06 (s, 1H), 7.96-7.94 (m, 2H), 7.47-7.35 (m, 3H), 6.76 (d, J = 8.4 Hz, 1H), 4.90 (s, 1H), 4.67 (s, 1H), 4.25-4.22 (m, 1H), 4.05-3.90 (m, 3H), 3.99 (s, 3H), 3.21-3.18 (m, 1H), 3.01-2.98 (m, 1H), 1.96-1.81 (m, 4H).

synthesis procedures for examples 289 to 320 Synthesis with reference to example 288: wherein 2- (5-fluoropyridin-2-yl) acetic acid is substituted with a different acid:

。

the synthesis procedure of examples 321 to 322 was synthesized with reference to example 286.

The synthesis procedure of examples 323 to 324 was synthesized with reference to example 288.

Synthetic procedure of examples 325 to 365 were synthesized with reference to example 183.

Example 366

3-Chloro-N- (4- (hydroxymethyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) -2-carbamoyl pyridine

First step

(4- (Hydroxymethyl) -1- (5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester

Tert-butyl (1- (5-bromo-2-yl) -4- (hydroxymethyl) pyridin-2-yl) piperidin-4-yl) carbamate (second step of synthesis reference example 44) (1.00 g,2.6 mmol) and pinacol diboronate (0.99 g,3.9 mmol) were dissolved in dioxane (30 mL), bis (triphenylphosphine) palladium dichloride (0.19 g,0.26 mmol) and potassium acetate (0.51 g,5.2 mmol) were added in sequence and reacted at 90℃under nitrogen protection for 8 hours. After the reaction solution is cooled to room temperature, the crude dioxane solution is directly fed to the next step. MS M/z (ESI) 434 [ M+1 ];

Second step

(1- (5- (7-Chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (hydroxymethyl) piperidin-4-yl) carbamic acid tert-butyl ester

To a solution of tert-butyl (4- (hydroxymethyl) -1- (5- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridin-2-yl) piperidin-4-yl) carbamate (1.14 g,2.6 mmol) in dioxane (30 mL) was added sequentially 5, 7-dichloro-1, 6-naphthyridine (0.52 g,2.6 mmol), tetrakis (triphenylphosphine) palladium (0.30 g,0.26 mmol), potassium carbonate (0.90 g,6.5 mmol) and water (6 mL). Heating to 85 ℃ under the protection of nitrogen, and reacting for 16 hours. After the reaction solution was cooled to room temperature, ethyl acetate (50 mL) and water (20: 20 mL) were added to dilute the mixture, and the organic phase was separated, and the aqueous phase was extracted with ethyl acetate (50 mL ×2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate) to give tert-butyl (1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (hydroxymethyl) piperidin-4-yl) carbamate (0.65 g, yellow solid) as the target compound in 53.2% yield. MS M/z (ESI): 470 & 472 [ M+1 ];

Third step

(4- (Hydroxymethyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester

Tert-butyl (1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (hydroxymethyl) piperidin-4-yl) carbamate (0.65 g,1.39 mmol), 1-methyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (0.58 g,2.8 mmol) was dissolved in N, N-dimethylacetamide (10 mL), and tetrakis (triphenylphosphine) palladium (0.16 g,0.14 mmol), potassium phosphate (0.89 g,4.17 mmol) and water (2 mL) were added sequentially. The reaction was heated to 140℃under nitrogen for 4 hours. The reaction mixture was cooled to room temperature, poured into water (50 mL), and extracted with ethyl acetate (50 mL ×3). The organic phases were combined and washed with saturated brine (50 mL ×3), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. Purification of the residue by column chromatography (petroleum ether: ethyl acetate=1:3) afforded the target compound tert-butyl (4- (hydroxymethyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) carbamate (0.42 g, white solid), yield 58.8%;

MS m/z (ESI): 516 [M + 1];

¹H NMR (400 MHz, CDCl₃) δ 8.98-8.97 (m, 1H), 8.56 (d, J = 2.4 Hz, 1H), 8.41 (d, J = 8.0 Hz, 1H), 8.12 (s, 1H), 8.09 (s, 1H), 7.93-7.92 (m, 2H), 7.37-7.33 (m, 1H), 6.83 (d, J = 8.8 Hz, 1H), 4.98 (s, 1H), 4.06-4.01 (m, 2H), 3.97 (s, 3H), 3.77 (s, 2H), 3.40-3.35 (m, 2H), 2.14-2.10 (m, 2H), 1.83-1.76 (m, 2H), 1.45 (s, 9H);

Fourth step

4- (Hydroxymethyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-amine

(4- (Hydroxymethyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester (0.28 g,0.54 mmol) was dissolved in methanol (3 mL), dioxane hydrochloride solution (3 mL,12.00 mmol,4M) was added, and the reaction was carried out at room temperature for 3 hours. The reaction solution was concentrated, dichloromethane (15 mL) was added, the pH was adjusted to 8-9 with triethylamine, and concentrated again to give crude 4- (hydroxymethyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-amine (crude, 0.45 g, yellow solid) as the target compound, which was directly fed to the next step. MS M/z (ESI): 416 [ M+1 ];

Fifth step

3-Chloro-picolinic acid (10 mg,0.06 mmol), 2- (7-Oxyltriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (27 mg,0.072 mmol), triethylamine (20 mg,0.20 mmol) were dissolved in N, N-dimethylformamide (4 mL), stirred at room temperature for 10 minutes, 4- (hydroxymethyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-amine (crude product, 80 mg,0.096 mmol) was added and the reaction was stirred at room temperature for 2 hours. The reaction solution was concentrated, methylene chloride (20 mL) was added, and washed successively with 1M sodium hydroxide solution (5 mL,5 mmol) and saturated brine (25 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Purification of the residue by high performance liquid chromatography gave the title compound 3-chloro-N- (4- (hydroxymethyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) -2-carbamoyl pyridine (28 mg, yellow solid) in 52.6% yield;

MS m/z (ESI): 555 [M + 1];

¹H NMR (400 MHz, CDCl₃) δ 9.02-9.01 (m, 1H), 8.59 (d, J = 2.4 Hz, 1H), 8.47-8.44 (m, 2H), 8.17 (s, 1H), 8.12 (s, 1H), 8.08 (s, 1H), 7.99-7.96 (m, 2H), 7.86-7.83 (m, 1H), 7.42-7.38 (m, 2H), 6.88 (d, J = 8.8 Hz, 1H), 4.16-4.12 (m, 2H), 3.99 (s, 3H), 3.94 (s, 2H), 3.51-3.46 (m, 2H), 2.35-2.31 (m, 2H), 1.95-1.89 (m, 2H).

Synthesis procedures of examples 367 to 371 were followed for synthesis in accordance with example 366.

The synthesis of example 372 is described with reference to the procedure of example 44.

Example 373

3-Fluoro-N- (1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- ((morpholin-4-yl) methyl) piperidin-4-yl) -2-carbamoyl pyridine

First step

(6- (4- ((Tert-Butoxycarbonyl) amino) -4- (methoxycarbonyl) piperidin-1-yl) pyridin-3-yl) boronic acid

The compound methyl 1- (5-bromopyridin-2-yl) -4- ((tert-butoxycarbonyl) amino) piperidine-4-carboxylate (first step of synthesis reference example 44) (4.60 g,11.14 mmol) was added to 1, 4-dioxane (50 mL), pinacol bisborate (4.24 g,16.71 mmol), 1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (1.22 g, 1.67 mmol) and potassium acetate (3.27 g, 33.41 mmol) were added in portions. After the addition was completed, argon was replaced three times, magnetically stirred at 90 ℃ and condensed and refluxed for 3 hours. Cooled to room temperature and desolventized under reduced pressure to give the target product (6- (4- ((tert-butoxycarbonyl) amino) -4- (methoxycarbonyl) piperidin-1-yl) pyridin-3-yl) boronic acid (crude), which was used in the next reaction without purification. MS m/z (ESI): 380 [ M+1 ];

Second step

4- ((Tert-Butoxycarbonyl) amino) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidine-4-carboxylic acid methyl ester

The compound (6- (4- ((tert-butoxycarbonyl) amino) -4- (methoxycarbonyl) piperidin-1-yl) pyridin-3-yl) boronic acid (crude, 11.14 mmol), 5, 7-dichloro-1, 6-naphthyridine (2.65 g,13.37 mmol), tetrakis (triphenylphosphine) palladium (1.92 g,1.67 mmol), anhydrous potassium carbonate (3.07 g,22.28 mmol), 1, 4-dioxane (50 mL) and water (5 mL) were added in a 250 mL single vial with a mixture of three argon substitutions, magnetically stirred at 120 ℃ and condensed back for 16 hours. After cooling to room temperature, 100 mL water quench was added, extraction was performed with ethyl acetate (100 mL ×3), the organic phases were combined and washed with saturated brine (100 mL ×2), dried over anhydrous sodium sulfate, the drying agent was removed by filtration, the filtrate was desolventized under reduced pressure, and the residue was purified by chromatography on a silica gel column (petroleum ether/ethyl acetate=100:1-7:3) to give the target product 4- ((tert-butoxycarbonyl) amino) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidine-4-carboxylic acid methyl ester (3.20 g, yellow oil). Yield: 58 The%;

MS m/z (ESI): 498&500 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.08-9.06 (m, 1H), 8.53-8.47 (m, 2H), 7.95 (m, 1H), 7.91 (s, 1H), 7.18 (s, 1H), 6.84 (d, J = 8.9 Hz, 1H), 4.85 (s, 1H), 4.14-4.12 (m, 4H), 3.76 (s, 3H), 3.44-3.42 (m, 2H), 2.21-3.19 (m, 2H), 1.46 (s, 9H);

Third step

4- ((Tert-Butoxycarbonyl) amino) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidine-4-carboxylic acid methyl ester

Methyl compound 4- ((tert-butoxycarbonyl) amino) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidine-4-carboxylate (3.20 g,6.43 mmol), pinacol 1-methyl-4-pyrazole borate (1.6 g, 7.71 mmol), tetrakis (triphenylphosphine) palladium (1.11 g,0.96 mmol), anhydrous potassium carbonate (1.77 g,12.85 mmol), N-dimethylacetamide (50 mL) were added to a 250mL single-necked flask, replaced with argon three times, magnetically stirred at 120 ℃ and condensed reflux for 8 hours. After cooling to room temperature, 100mL water quench was added, extraction was performed with ethyl acetate (100 mL ×3), the organic phases were combined and washed with saturated brine (100 mL ×2), dried over anhydrous sodium sulfate, the drying agent was removed by filtration, the filtrate was desolventized under reduced pressure, and the residue was purified by chromatography on a silica gel column (dichloromethane/methanol=100:1 to 9:1) to give the title product 4- ((tert-butoxycarbonyl) amino) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidine-4-carboxylic acid methyl ester (2.77 g, yellow oil). Yield: 79 The%;

MS m/z (ESI): 544 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.02-9.00 (m, 1H), 8.58 (d, J = 2.3 Hz, 1H), 8.43 (d, J = 8.5 Hz, 1H), 8.12 (s, 1H), 8.08 (s, 1H), 7.98-7.96 (m, 2H), 7.38-7.36 (m, 1H), 6.86 (d, J = 8.8 Hz, 1H), 4.91 (s, 1H), 4.18-4.09 (m, 2H), 3.99 (s, 3H), 3.76 (s, 3H), 3.55-3.34 (m, 4H), 2.26-2.18 (m, 2H), 1.46 (s, 9H);

Fourth step

The compound methyl 4- ((tert-butoxycarbonyl) amino) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidine-4-carboxylate (1.00 g,1.842 mmol) was added to tetrahydrofuran (10 mL), then lithium aluminum hydride (70 mg,1.841 mmol) was added slowly in portions at 0 ℃ and magnetically stirred at 0 ℃ for 0.5 hours. Quench with water (30 mL), extract with ethyl acetate (30 mL x 3), combine the organic phases and wash with saturated brine (100 mL x 2), dry over anhydrous sodium sulfate and filter to remove the desiccant. The filtrate was desolventized under reduced pressure and the residue was purified by chromatography on a silica gel column (dichloromethane/methanol=100:1 to 9:1) to give the target product tert-butyl (4- (hydroxymethyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamate (0.70 g, yellow solid). Yield: 74 The%;

MS m/z (ESI): 516 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.02-9.00 (m, 1H), 8.59 (d, J = 2.2 Hz, 1H), 8.43 (d, J = 8.4 Hz, 1H), 8.16-8.05 (m, 2H), 8.00-7.87 (m, 2H), 7.39-7.32 (m, 1H), 6.86 (d, J = 8.8 Hz, 1H), 4.68 (s, 1H), 4.03-4.01 (m, 2H), 3.99 (s, 3H), 3.78 (s, 2H), 3.49-3.36 (m, 2H), 2.12-1.99 (m, 2H), 1.86-1.75 (m, 2H), 1.46 (s, 9H);

Fifth step

(4-Formyl-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester

The compound (4- (hydroxymethyl) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester (0.70 g,1.359 mmol) was added to dichloromethane (10 mL), followed by the addition of dessert-martin oxidant (69 mg,1.631 mmol) and magnetic stirring at room temperature for 1 hour. Quenched by addition of saturated aqueous sodium sulfite (15 mL), followed by addition of saturated aqueous sodium bicarbonate (35 mL) and extraction with ethyl acetate (50 mL ×3). The organic phases were combined and washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was desolventized under reduced pressure to give tert-butyl (4-formyl-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamate (0.60 g, yellow oil) as the target product. Yield: 86 The%;

MS m/z (ESI): 514 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.56 (s, 1H), 9.03-8.98 (m, 1H), 8.59-8.57 (m, 1H), 8.43 (d, J = 8.4 Hz, 1H), 8.11-8.09 (m, 2H), 8.00-7.95 (m, 2H), 7.39-7.33 (m, 1H), 6.86 (d, J = 8.9 Hz, 1H), 4.66 (s, 1H), 4.20-4.06 (m, 2H), 3.99 (s, 3H), 3.21-3.18 (m, 2H), 2.14-2.02 (m, 2H), 1.97-1.85 (m, 2H), 1.43 (s, 9H);

Sixth step

(7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- ((morpholin-4-yl) methyl) piperidin-4-yl) carbamic acid tert-butyl ester

The compound (4-formyl-1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) piperidin-4-yl) carbamic acid tert-butyl ester (0.12 g,1.167 mmol) was dissolved in 1, 2-dichloroethane (10 mL), and morpholine (0.15 g,1.400 mmol), zinc chloride (0.79 g,5.835 mmol) and sodium cyanoborohydride (0.15 g,2.334 mmol) were added in this order and stirred at room temperature for 3 hours. Quench with water (30 mL), extract with dichloromethane (30 mL x 3), combine the organic phases and wash with saturated brine (100 mL x 2), dry over anhydrous sodium sulfate and filter to remove the desiccant. The filtrate was desolventized under reduced pressure and the residue was purified by chromatography on a silica gel column (dichloromethane/methanol=100:1-9:1) to give the target product tert-butyl (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) -4- ((morpholin-4-yl) methyl) piperidin-4-yl) carbamate (0.30 g, yellow oil). Yield: 44 The%;

MS m/z (ESI): 585 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.02-9.00 (m, 1H), 8.58 (d, J = 2.2 Hz, 1H), 8.44 (d, J = 8.0 Hz, 1H), 8.11-8.09 (m, 2H), 7.99-7.92 (m, 2H), 7.39-7.27 (m, 1H), 6.84 (d, J = 8.8 Hz, 1H), 4.61 (s, 1H), 4.13-4.11 (m, 4H), 3.99 (s, 3H), 3.70-3.68 (m, 4H), 3.25-3.23 (m, , 4H), 2.86-2.84 (m, 2H), 2.60-2.55 (m, 4H), 1.45 (s, 9H);

Seventh step

1- (5- (7- (1-Methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- ((morpholin-4-yl) methyl) piperidin-4-amine hydrochloride

Tert-butyl (15 mg,0.03 mmol) of the compound (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- ((morpholin-4-yl) methyl) piperidin-4-yl) carbamate (15 mg,0.03 mmol) was dissolved in dichloromethane (1 mL) and methanol (0.5 mL), dioxane hydrochloride (1 mL,4.00 mmol,4M) was added at room temperature, and the reaction was stirred at 25℃for 30 minutes. The reaction solution is decompressed and desolventized to obtain crude 1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridine-2-yl) -4- ((morpholin-4-yl) methyl) piperidine-4-amine hydrochloride, and the next reaction is directly carried out. MS M/z (ESI) 485 [ M+1 ];

eighth step

The compound 1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- ((morpholin-4-yl) methyl) piperidin-4-amine hydrochloride (crude, 0.03 mmol), 3-fluoropyridine-2-carboxylic acid (7 mg,0.036 mmol) and triethylamine (10 mg,0.09 mmol) were dissolved in N, N-dimethylformamide (2 mL), and 2- (7-benzotriazol-on-N, N' -tetramethylurea hexafluorophosphate (18 mg,0.045 mmol) was added at room temperature and magnetically stirred at 25 ℃ for 1 hour. Quench with water (20 mL) and extract with dichloromethane (30 mL x 3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to preparative high performance liquid chromatography to give the target product 3-fluoro-N- (1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- ((morpholin-4-yl) methyl) piperidin-4-yl) -2-carbamoyl pyridine (6.8 mg, yellow solid). Yield: 38 The%;

MS m/z (ESI): 608 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.03-9.01 (m, 1H), 8.59 (d, J = 2.2 Hz, 1H), 8.47-8.39 (m, 2H), 8.14-8.05 (m, 2H), 7.96 (s, 2H), 7.61-7.55 (m, 1H), 7.52 (s, 1H), 7.39-7.37 (m, 1H), 6.87 (d, J = 8.8 Hz, 1H), 4.29 (s, 2H), 3.99 (s, 3H), 3.78-3.76 (m, 4H), 3.32-3.30 (m, 2H), 3.12-3.10 (m, 2H), 2.85-2.83 (m, 4H), 2.64-2.62 (m, 2H), 1.84-1.82 (m, 2H).

The synthesis procedure of examples 374 to 412 refers to the synthesis of example 373.

Example 446

7- (1-Methyl-1H-pyrazol-4-yl) -5- (6- (4- (methylsulfonyl) piperidin-1-yl) pyridin-3-yl) -1, 6-naphthyridine

First step

5-Bromo-2- (4- (methylsulfonyl) piperidin-1-yl) pyridine

The compound 5-bromo-2-fluoropyridine (0.89 g,5.06 mmol), 4- (methylsulfonyl) piperidine (0.75 g,4.60 mmol), potassium carbonate (1.27 g,9.20 mmol) was dissolved in dimethyl sulfoxide (10 mL), reacted at 80℃for 8 hours, the reaction solution was poured into water (50 mL) and extracted with ethyl acetate (30 mL X3). The combined organic phases were washed with water (50 mL ×3) and saturated brine (50 mL ×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by chromatography on a silica gel column (petroleum ether/ethyl acetate=100:1 to 3:1) to give the target product 5-bromo-2- (4- (methylsulfonyl) piperidin-1-yl) pyridine (0.62 g, yellow solid). Yield: 42 The%;

MS m/z (ESI)：319&321 [M + 1];

¹H NMR (400 MHz, CDCl₃) δ 8.19 (d, J = 2.4 Hz, 1H), 7.56-7.53 (m, 1H), 6.59 (d, J = 9.0 Hz, 1H), 4.46-4.44 (m, 2H), 3.12-3.02 (m, 1H), 2.93-2.86 (m, 2H), 2.85 (s, 3H), 2.24-2.20 (m, 2H), 1.90-1.79 (m, 2H);

Second step

(6- (4- (Methylsulfonyl) piperidin-1-yl) pyridin-3-yl) boronic acid

The compound 5-bromo-2- (4- (methylsulfonyl) piperidin-1-yl) pyridine (0.62 g,1.94 mmol), pinacol biborate (0.99 g,3.89 mmol), potassium acetate (0.38 g,3.88 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride dichloromethane complex (15 mg,0.02 mmol) and 1, 4-dioxane (20 mL) were added to a single vial of 100 mL, after which the gases were replaced three times with argon and the reaction was allowed to react at 90℃for 4 hours. The crude product (1.94 mmol) of the target product (6- (4- (methylsulfonyl) piperidin-1-yl) pyridin-3-yl) boric acid obtained in this way was used directly in the next step without post-treatment. MS m/z (ESI): 285 [ M+1 ];

Third step

7-Chloro-5- (6- (4- (methylsulfonyl) piperidin-1-yl) pyridin-3-yl) -1, 6-naphthyridine

The compound (6- (4- (methylsulfonyl) piperidin-1-yl) pyridin-3-yl) boronic acid (crude, 1.94 mmol), 5, 7-dichloro-1, 6-naphthyridine (0.42 g,2.13 mmol), potassium carbonate (0.53 g,3.88 mmol), tetrakis (triphenylphosphine) palladium (23 mg,0.02 mmol) and water (4 mL) were added to a single vial of 100 mL, after which the gases were replaced three times with argon and the reaction was allowed to react at 100℃for 6 hours. Cooled to room temperature, diluted with water (50 mL) and extracted with ethyl acetate (50 mL ×3). The organic phases were combined and washed with brine (50 mL ×2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was purified by chromatography on silica gel (dichloromethane/methanol=100:1 to 25:1) to give the target product 7-chloro-5- (6- (4- (methylsulfonyl) piperidin-1-yl) pyridin-3-yl) -1, 6-naphthyridine (0.40 g, yellow solid). Yield: 51 Percent of the total weight of the composition. MS M/z (ESI): 403 & 405 [ M+1 ];

Fourth step

The compound 7-chloro-5- (6- (4- (methylsulfonyl) piperidin-1-yl) pyridin-3-yl) -1, 6-naphthyridine (20 mg,0.05 mmol), 1-methyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (13 mg,0.06 mmol), potassium phosphate (22 mg,0.10 mmol), tetrakis (triphenylphosphine) palladium (12 mg,0.01 mmol) and N, N-dimethylacetamide (10 mL)/water (2 mL) were mixed, replaced three times with argon and the reaction was allowed to react at 120℃for 2 hours. Dilute with water (50 mL) and extract with ethyl acetate (50 mL x 3). The organic phase was washed with water (50 mL ×3) and saturated brine (50 mL ×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative liquid phase (Agilent ZORBAX XDB-C18, 4.6x50 mm,3.5 μm, ACN/H ₂ O ((0.1% TFA) 15% -40%) to give the desired product 7- (1-methyl-1H-pyrazol-4-yl) -5- (6- (4- (methylsulfonyl) piperidin-1-yl) pyridin-3-yl) -1, 6-naphthyridine (6.80 mg, yellow solid): 31%;

MS m/z (ESI): 449 [M + 1];

¹H NMR (400 MHz, CDCl₃) δ 9.02-9.01 (m, 1H), 8.59-8.58 (m, 1H), 8.43 (d, J = 8.4 Hz, 1H), 8.12 (s, 1H), 8.08 (s, 1H), 7.99-7.97 (m, 1H), 7.97 (s, 1H), 7.40-7.37 (m, 1H), 6.87 (d, J = 8.8 Hz, 1H), 4.69-4.67 (m, 2H), 3.99 (s, 3H), 3.21-3.13 (m, 1H), 3.07-3.00 (m, 2H), 2.89 (s, 3H), 2.29-2.26 (m, 2H), 1.98-1.90 (m, 2H).

the synthesis procedure of examples 447 to 448 refers to the synthesis of example 446.

Example 449

(3S, 4S) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-amine

First step

(3S, 4S) -3- (1, 3-Dioxoisoindolin-2-yl) -4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester

Tert-butyl (3S, 4S) -3-amino-4-hydroxypyrrolidine-1-carboxylate (0.25 g,1.20 mmol), phthalic anhydride (0.18 g,1.20 mmol), triethylamine (0.36 g,3.60 mmol) and N, N-4-dimethylaminopyridine (12 mg,0.10 mmol) were added to tetrahydrofuran (10 mL), and the mixture was warmed to 70℃and reacted overnight. The residue was dissolved in ethyl acetate (50 mL) and washed with saturated brine (50 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column (30-50% ethyl acetate/petroleum ether) to give the desired product (3 s,4 s) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-amine (0.15 g, yellow solid). Yield: 36.5 The%;

MS m/z (ESI)：355 [M + 23]；

¹H NMR (400 MHz, CDCl₃) δ 7.88-7.83 (m, 2H), 7.78-7.72 (m, 2H), 5.08-4.94 (m, 1.5H), 4.66-4.56 (m, 1.5H), 4.02-3.80 (m, 3H), 2.23-2.21 (m, 1H), 1.47 (s, 9H);

Second step

2- (((3S, 4S) -1- (tert-Butoxycarbonyl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) carbamoyl) benzoic acid

(3S, 4S) -3-amino-4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester (0.15 g,0.45 mmol) was dissolved in anhydrous N, N-dimethylacetamide (2 mL), sodium hydrogen (60% dispersed in mineral oil, 54 mg,1.35 mmol) was added. Stirring was carried out at room temperature for half an hour, then 2-fluoropyridine (0.13 g,1.35 mmol) was added, and the temperature was raised to 80℃to react for 4 hours. After cooling to room temperature, the reaction was also poured into water (20 mL), and the aqueous phase was extracted with ethyl acetate (15 mL ×3). The organic phases were combined, washed with saturated brine (30 mL ×2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to give the desired product 2- (((3 s,4 s) -1- (tert-butoxycarbonyl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) carbamoyl) benzoic acid (0.20: 0.20 g, black oil). Crude product. MS m/z (ESI): 428 [ M+1 ];

Third step

2- ((3S, 4S) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) isoindoline-1, 3-dione hydrochloride

2- (((3S, 4S) -1- (tert-Butoxycarbonyl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) carbamoyl) benzoic acid (0.20 g,0.45 mmol) was dissolved in a solution of hydrogen chloride in methanol (5 mL,20.00 mmol,4M) and stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to give the desired product 2- ((3 s,4 s) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) isoindoline-1, 3-dione hydrochloride (crude, 0.45 mmol). Crude product. MS m/z (ESI): 310 [ M+1 ];

Fourth step

2- ((3S, 4S) -1- (5-bromopyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) isoindoline-1, 3-dione

2- ((3S, 4S) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) isoindoline-1, 3-dione hydrochloride (crude, 0.45 mmol), 5-bromo-2-fluoropyridine (0.24 g,1.35 mmol) and N, N-diisopropylethylamine (0.35 g,2.70 mmol) were dissolved in dimethyl sulfoxide (1 mL), warmed to 100℃and stirred overnight. After cooling to room temperature, the reaction solution was poured into water (20 mL), and the aqueous phase was extracted with ethyl acetate (20 mL ×2). The organic phases were combined and washed with water (30 mL ×2) and saturated brine (30 mL ×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was desolventized under reduced pressure. The residue was purified by column chromatography on silica gel to give the desired product 2- ((3 s,4 s) -1- (5-bromopyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) isoindoline-1, 3-dione (65 mg, white solid). Yield: 31.1 The%;

MS m/z (ESI)：465 & 467 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 8.24-8.14 (m, 1H), 7.90-7.68 (m, 4H), 7.62-7.46 (m, 2H), 6.85-6.68 (m, 2H), 6.40-6.30 (m, 1H), 6.19-6.08 (m, 1H), 5.22-5.12 (m, 1H), 4.38-4.28 (m, 1H), 4.14-3.84 (m, 2H), 3.69-3.58 (m, 1H), 3.52-3.41 (m, 1H);

Fifth step

2- (((3S, 4S) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) carbamoyl) benzoic acid

The compound 2- ((3 s,4 s) -1- (5-bromopyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) isoindoline-1, 3-dione (65 mg,0.14 mmol), pinacol biborate (53 mg,0.21 mmol), potassium acetate (28 mg,0.28 mmol), 1' -bis-diphenylphosphino ferrocene palladium dichloride (7 mg,0.10 mmol) was added to dioxane (5 mL), purged three times with nitrogen, and the reaction was warmed to 90 ℃ for 4 hours. After cooling to room temperature, 5, 7-dichloro-1, 6-naphthyridine (28 mg,0.14 mmol), tetrakis triphenylphosphine palladium (12 mg,0.01 mmol), anhydrous potassium carbonate (38 mg,0.28 mmol) and water (1 mL) were added. The temperature was raised to 85 ℃ and reacted overnight under nitrogen atmosphere. After cooling to room temperature, the product 2- (((3 s,4 s) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) carbamoyl) benzoic acid (0.20 g) is obtained by concentration under reduced pressure. Crude product. MS M/z (ESI): 567 & 569 [ M+1 ];

Sixth step

2- (((3 S,4 s) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) carbamoyl) benzoic acid

A mixed solution of compound 2- (((3S, 4S) -1- (5- (7-chloro-1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) carbamoyl) benzoic acid (0.20 g,0.14 mmol), 1-methyl-4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (58 mg,0.28 mmol), potassium phosphate (59 mg,0.28 mmol) and tetrakis (triphenylphosphine) palladium (12 mg,0.01 mmol) was added, the mixture was purged three times with nitrogen, and the temperature was raised to 140℃for reaction for 4 hours. After cooling to room temperature, the reaction solution was diluted with water (20 mL), and the aqueous phase was washed with dichloromethane (20 mL ×2). The resulting aqueous phase was then concentrated under reduced pressure to give the product 2- (((3 s,4 s) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) carbamoyl) benzoic acid (0.23 g, black oil). Crude products; MS M/z (ESI) 613 [ M+1 ];

Seventh step

The compound 2- (((3S, 4S) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-yl) carbamoyl) benzoic acid (0.23 g,0.14 mmol) was dissolved in aqueous hydrochloric acid (6 mL,18.00 mmol,3M), and the mixture was stirred at 50℃for 3 hours. Concentrated under reduced pressure and the residue purified by preparative liquid phase (Agilent ZORBAX XDB-C18, 4.6X150 mm,3.5 μm, ACN/H ₂ O (0.5% HCOOH) 25% -40%) to give (3S, 4S) -1- (5- (7- (1-methyl-1H-pyrazol-4-yl) -1, 6-naphthyridin-5-yl) pyridin-2-yl) -4- (pyridin-2-yloxy) pyrrolidin-3-amine hydrochloride (1.8 mg, yellow solid), yield: 2.77 The%;

MS m/z (ESI): 465 [M + 1]；

¹H NMR (400 MHz, CDCl₃) δ 9.04-8.98 (m, 1H), 8.60-8.56 (m, 1H), 8.44 (d, J = 8.0 Hz, 1H), 8.20-8.16 (m, 1H), 8.12 (s, 1H), 8.07 (s, 1H), 8.00-7.89 (m, 2H), 7.63-7.56 (m, 1H), 7.42-7.33 (m, 1H), 6.94-6.88 (m, 1H), 6.76 (d, J= 8.0 Hz, 1H), 6.59 (d, J = 8.0 Hz, 1H), 5.42-4.35 (m, 1H), 4.24-4.14 (m, 1H), 4.05-3.85 (m, 5H), 3.77-3.69 (m, 1H), 3.58-3.52 (m, 1H).

The synthesis procedure of examples 450 to 451 was referred to the synthesis of example 449:

。

synthesis of example 461 reference example 223 Synthesis procedure.

Biological experiments

Activity inhibition assay for RET kinase

Evaluation of the Effect of the Compounds of the invention on RET kinase Activity Using in vitro kinase assay experiments

The experimental procedure is summarized as follows:

The in vitro activity of RET kinase was determined by detecting the level of phosphorylation of the substrate in the kinase reaction using a Homogeneous Time Resolved Fluorescence (HTRF) kinase assay kit (Cisbio, cat. 62TK0 PEC). The reaction buffer contained the following components: the kit is provided with enzyme reaction buffer (1×), 5mM MgCl ₂ and 1mM DTT; human recombinant RET protein (cat 11997) was purchased from Yinqiao Shenzhou and diluted with reaction buffer to 0.1 ng/. Mu.l kinase solution; the substrate reaction solution includes a biotin-labeled tyrosine kinase substrate diluted to 0.66. Mu.M with a reaction buffer and 12. Mu.M ATP; the detection buffer included Eu3+ labeled caged antibody diluted to 0.1 ng/. Mu.l with reaction buffer-and 41.25nM streptavidin labeled XL665.

Compounds were diluted to 25 μm in 100% DMSO, then serially diluted 4-fold with DMSO to a minimum concentration of 1.5nM, and diluted 40-fold with reaction buffer at each concentration point.

To 384-well assay plates (Corning, cat No. 4512) 4 μl of compound solution and 2 μl of RET kinase solution were added, mixed well and incubated at room temperature for 15 minutes. Subsequently, 4. Mu.L of the substrate reaction solution was added, and the reaction mixture was incubated at room temperature for 60 minutes. Then, 10. Mu.L of detection buffer was added in an equal volume to the reaction, and after mixing well and standing at room temperature for 30 minutes, the progress of the reaction was detected at 620nm and 665nm wavelengths using an Envision reader (PERKIN ELMER). 665/620 and the degree of phosphorylation of the substrate, whereby RET kinase activity is detected. In this experiment, the group without RET kinase protein was taken as 100% inhibition, the RET kinase protein was added but the group without compound was taken as 0% inhibition. The percent inhibition of RET kinase activity by a compound can be calculated by the following formula:

Compound IC ₅₀ values were calculated from 8 concentration points using XLfit (ID Business Solutions ltd., UK) software by the following formula:

Y = Bottom+(Top-Bottom)/(1+10^((logIC₅₀-X)*slope factor))

Wherein Y is the inhibition percentage, X is the logarithmic value of the concentration of the compound to be detected, bottom is the maximum inhibition percentage, top is the minimum inhibition percentage, and slope factor is the curve slope coefficient.

Test for inhibition of RET M918T Activity

Evaluation of the Effect of the Compounds of the invention on RET M918T Activity Using in vitro kinase assay experiments

The experimental procedure is summarized as follows:

The in vitro activity of RET M918T was determined by detecting the level of phosphorylation of the substrate in the kinase reaction using the HTRF kinase assay kit (Cisbio, cat. No. 62TK0 PEC). The reaction buffer contained the following components: the kit is provided with enzyme reaction buffer (1×), 5mM MgCl ₂ and 1mM DTT; human recombinant RET M918T protein (accession number PV 6217) was purchased from Thermo Fish company and diluted with reaction buffer to 0.35 ng/. Mu.l kinase solution; the substrate reaction solution includes a biotin-labeled tyrosine kinase substrate diluted to 0.9. Mu.M with a reaction buffer and 18. Mu.M ATP; the detection buffer included Eu ³⁺ -labeled cage antibody diluted to 0.1 ng/. Mu.l with reaction buffer and streptavidin-labeled XL665 at 56.25 nM.

Compounds were diluted to 10 μm in 100% DMSO, then serially diluted 4-fold with DMSO to a minimum concentration of 0.61nM, and diluted 40-fold with reaction buffer at each concentration point.

To 384-well assay plates (Corning, cat No. 4512) 4 μl of compound solution and 2 μl of RET M918T kinase solution were added, mixed well and incubated at room temperature for 15 minutes. Subsequently, 4. Mu.L of the substrate reaction solution was added, and the reaction mixture was incubated at room temperature for 40 minutes. Then, 10. Mu.L of detection buffer was added in an equal volume to the reaction, and after mixing well and standing at room temperature for 30 minutes, the progress of the reaction was detected at 620nm and 665nm wavelengths using an Envision reader (PERKIN ELMER). 665/620 and the degree of phosphorylation of the substrate, thereby detecting RET M918T kinase activity. In this experiment, the RET M918T kinase protein group was not added as 100% inhibition, the RET M918T kinase protein was added but the compound group was not added as 0% inhibition. The percent inhibition of RET M918T activity by a compound can be calculated by the following formula:

percent inhibition = 100-100 x (signal value at specific concentration of test compound-negative control signal value)/(positive control signal value-negative control signal value).

Y = Bottom+(Top-Bottom)/(1+10^((logIC₅₀-X)*slope factor))

Activity inhibition assay for RET V804M

Evaluation of the Effect of the Compounds of the invention on RET V804M Activity Using in vitro kinase assay experiments

The experimental procedure is summarized as follows:

the in vitro activity of RET V804M was determined by detecting the level of phosphorylation of the substrate in the kinase reaction using a Homogeneous Time Resolved Fluorescence (HTRF) kinase assay kit (Cisbio, cat No. 62TK0 PEC). The reaction buffer contained the following components: the kit is provided with enzyme reaction buffer (1×), 5mM MgCl ₂, 1mM DTT and 0.08% Tween-20; the recombinant RET V804M protein (accession number PV 6223) was purchased from Thermo Fish and diluted with reaction buffer to a kinase solution of 0.15 ng/. Mu.l; the substrate reaction solution includes a biotin-labeled tyrosine kinase substrate diluted to 0.9. Mu.M with a reaction buffer and 8. Mu.M ATP; the detection buffer included Eu ³⁺ -labeled cage antibody diluted to 0.1 ng/. Mu.l with reaction buffer and streptavidin-labeled XL665 at 56.25 nM.

To 384-well assay plates (Corning, cat No. 4512) were added 4 μl of compound solution and 2 μl of RET V804M kinase solution, and after mixing well, incubated at room temperature for 15 minutes. Subsequently, 4. Mu.L of the substrate reaction solution was added, and the reaction mixture was incubated at room temperature for 30 minutes. Then, 10. Mu.L of detection buffer was added in an equal volume to the reaction, and after mixing well and standing at room temperature for 30 minutes, the progress of the reaction was detected at 620nm and 665nm wavelengths using an Envision reader (PERKIN ELMER). 665/620 and the degree of phosphorylation of the substrate, whereby RET V804M kinase activity was detected. In this experiment, the RET V804M kinase protein group was not added as 100% inhibition, the RET V804M kinase protein was added but the compound group was not added as 0% inhibition. The percent inhibition of RET V804M activity by a compound can be calculated by the following formula:

Y = Bottom+(Top-Bottom)/(1+10^((logIC₅₀-X)*slope factor))

Activity inhibition test of RET V804L

Evaluation of the Effect of the Compounds of the invention on RET V804L Activity Using in vitro kinase assay experiments

The experimental procedure is summarized as follows:

The in vitro activity of RET V804L was determined by detecting the level of phosphorylation of the substrate in the kinase reaction using the HTRF kinase assay kit (Cisbio, cat. No. 62TK0 PEC). The reaction buffer contained the following components: the kit is provided with enzyme reaction buffer (1×), 5mM MgCl ₂, 1mM DTT and 0.05% Tween-20; human recombinant RET V804L protein (cat No. 14-758) was purchased from Merck and diluted with reaction buffer to 0.05 ng/. Mu.l kinase solution; the substrate reaction solution includes 1. Mu.M biotin-labeled tyrosine kinase substrate and 8. Mu.M ATP diluted with reaction buffer; the detection buffer included Eu ³⁺ labeled cage antibody diluted to 0.1 ng/. Mu.l with reaction buffer and 62.5nM streptavidin labeled XL665.

Compounds were diluted to 100 μm in 100% DMSO, then serially diluted 4-fold with DMSO to a minimum concentration of 6.1nM, and diluted 40-fold with reaction buffer at each concentration point.

To 384-well assay plates (Corning, cat No. 4512) 4 μl of compound solution and 2 μl of RET V804L kinase solution were added, mixed well and incubated at room temperature for 15 minutes. Subsequently, 4. Mu.L of the substrate reaction solution was added, and the reaction mixture was incubated at room temperature for 40 minutes. Then, 10. Mu.L of detection buffer was added in an equal volume to the reaction, and after mixing well and standing at room temperature for 30 minutes, the progress of the reaction was detected at 620nm and 665nm wavelengths using an Envision reader (PERKIN ELMER). 665/620 and the degree of phosphorylation of the substrate, whereby RET V804L kinase activity was detected. In this experiment, the RET V804L kinase protein group was not added as 100% inhibition, the RET V804L kinase protein was added but the compound group was not added as 0% inhibition. The percent inhibition of RET V804L activity by a compound can be calculated by the following formula:

Y = Bottom+(Top-Bottom)/(1+10^((logIC₅₀-X)*slope factor))

Inhibition of VEGFR2 Activity test

Evaluation of the Effect of the Compounds of the invention on VEGFR2 Activity Using in vitro kinase assay experiments

The experimental procedure is summarized as follows:

the in vitro activity of VEGFR2 was determined by detecting the level of phosphorylation of the substrate in the kinase reaction using HTRF kinase assay kit (Cisbio, cat No. 62TK0 PEC). The reaction buffer contained the following components: the kit was provided with enzyme reaction buffer (1×), 5mM MgCl ₂、1mM DTT、1mM MnCl₂, 0.01% BSA and 0.05% Tween-20; human recombinant VEGFR2 protein (accession number 10012) was purchased from Yinqiao Shenzhou and diluted with reaction buffer to 0.3 ng/. Mu.l kinase solution; the substrate reaction solution includes a biotin-labeled tyrosine kinase substrate diluted to 0.3. Mu.M with a reaction buffer and 3.5. Mu.M ATP; the detection buffer included Eu ³⁺ -labeled cage antibody diluted to 0.1 ng/. Mu.l with reaction buffer and 18.75nM streptavidin-labeled XL665.

Compounds were diluted to 1000 μm in 100% DMSO, then serially diluted 4-fold with DMSO to a minimum concentration of 0.06 μm, and diluted 40-fold with reaction buffer at each concentration point.

To 384-well assay plates (Corning, cat No. 4512) were added 4 μl of compound solution and 2 μl of VEGFR2 kinase solution, and after mixing well, incubated at room temperature for 15 minutes. Subsequently, 4. Mu.L of the substrate reaction solution was added, and the reaction mixture was incubated at room temperature for 40 minutes. Then, 10. Mu.L of detection buffer was added in an equal volume to the reaction, and after mixing well and standing at room temperature for 30 minutes, the progress of the reaction was detected at 620nm and 665nm wavelengths using an Envision reader (PERKIN ELMER). 665/620 and the degree of phosphorylation of the substrate, whereby the activity of the VEGFR2 kinase is detected. In this experiment, the VEGFR2 kinase protein group was not added as 100% inhibition, the VEGFR2 kinase protein group was added but the compound group was not added as 0% inhibition. The percent inhibition of VEGFR2 activity by a compound can be calculated using the following formula:

percent inhibition = 100-100 × (signal at specific concentration of test compound-negative control signal)/(positive control signal-negative control signal)

Y = Bottom+(Top-Bottom)/(1+10^((logIC₅₀-X)*slope factor))

Determination of half-effective inhibitory concentration GI ₅₀ of Ba/F3 KIF5B-RET cells

The effect of the compounds of the invention on Ba/F3 KIF5B-RET cell proliferation was evaluated using a luminescent cell viability test experiment.

The experimental procedure is summarized as follows:

Using CELLTILTER-Glo (CTG) assay kit, the cell proliferation status of Ba/F3 KIF5B-RET was detected by detecting an indicator ATP of viable cell metabolism using a unique, stable luciferase, the luminescence signal generated in the assay being proportional to the number of viable cells in the medium.

Ba/F3 KIF5B-RET cells (CBP 73195, nanjac) were cultured in RPMI1640 medium (Thermofisher, 12440053) containing 10% FBS (GBICO, 10099-141) and 100units/ml Streptomyces lividans (Thermofisher, 15140122), and the cultured cells were grown in white 384-well plates (Thermofisher, 164610) after being digested and blown off with 0.25% pancreatin (Thermofisher, 25200056), with 1000 cells in 27. Mu.L of medium per well, and the 384-well plates were placed in a 5% CO ₂ -containing incubator at 37℃overnight. Compounds were dissolved in 100% DMSO and diluted to 5mM, after which 4-fold serial dilutions were made in DMSO to a minimum concentration of 0.31. Mu.M, and each concentration point was diluted 50-fold with RPMI1640 medium. If the compound GI ₅₀ value is very low, the initial concentration of the compound can be reduced. mu.L of diluted compound was added to each well and gently centrifuged to mix. Wherein, the culture medium without cells was used as a negative control (100% inhibition), and a 0.2% DMSO group was used as a positive control (0% inhibition). The 384 well plate was placed in an incubator at 37 ℃ with 5% CO ₂ for continuous incubation, after 72 hours, was removed and placed at room temperature for 30 minutes, CTG reagent was also removed and equilibrated to room temperature, 30 μ LCTG reagent was added to each well, placed on a shaker with gentle shaking to ensure adequate cell lysis, placed for 10 minutes to stabilize the luminescence signal, and then the luminescence signal was read with EnVision (Perkin Elmer). In addition, for correcting the cell number, a T ₀ control is simultaneously arranged, wherein the T ₀ control comprises a blank control only containing a culture medium and a cell adding control, the difference value of the blank control and the cell adding control is set as a T ₀ control, and the cell adding control is obtained by adding a CTG reagent before adding medicine.

The percent inhibition of Ba/F3 KIF5B-RET cell proliferation by a compound can be calculated using the following formula:

percent inhibition =100-100*{[(signal_{Compounds of formula (I)}-Signal_{Negative control})-T_{0 control}]/[(signal_{positive control}-Signal_{Negative control})_-T_{0 control}]}.

Compound GI ₅₀ values were calculated from 8 concentration points using XLfit (ID Business Solutions ltd., UK) software by the following formula:

Y = Bottom + (Top- Bottom)/(1+10^((LogIC₅₀-X) * slope factor))

where Y is the percent inhibition, bottom is the Bottom plateau of the curve (Bottom plateau of S-curve), top is the Top plateau of the curve (Top plateau of S-curve), and X is the log of the concentration of the test compound.

Determination of the half-potent inhibitory concentration GI ₅₀ of LC-2/ad cells

The effect of the compounds of the invention on LC-2/ad cell proliferation was evaluated using a luminescent cell viability test experiment.

The experimental procedure is summarized as follows:

Using CELLTILTER-Glo (CTG) assay kit, the cell proliferation status of LC-2/ad was detected by detecting the indicator ATP of viable cell metabolism using a unique, stable luciferase, the luminescence signal generated in the assay being proportional to the number of viable cells in the medium.

LC-2/ad cells (available from Shang Hai Yu biological Co.) were cultured in RPMI 1640:F12 (1:1) complete medium (Thermofisher, 72400047, 11765054) containing 10% FBS (GBICO, 10099-141) and 100units/ml Streptomyces lividans mix (Thermofisher, 15140122), when the coverage of cells in the culture vessel reached 80-90%, digested and blown off with 0.25% pancreatin (containing EDTA) (Thermofisher, 25200056) and planted in white 384-well plates (Thermofisher, 164610) with 27. Mu.l IMDM complete medium containing 1000 cells per well, and the 384-well plates were placed in a 37℃incubator containing 5% CO ₂ overnight. Compounds were dissolved in 100% DMSO and diluted to 1mM, after which 4-fold serial dilutions were made in DMSO to a minimum concentration of 0.061. Mu.M, and 50-fold dilutions were made at each concentration point using RPMI 1640:F12 (1:1) medium. If the compound GI ₅₀ value is very low, the initial concentration of the compound can be reduced. Mu.l of diluted compound was added to each well and gently centrifuged to mix. Wherein, the culture medium without cells was used as a negative control (100% inhibition), and a 0.2% DMSO group was used as a positive control (0% inhibition). The 384 well plate was placed in an incubator at 37℃with 5% CO ₂ for continuous incubation, after 96 hours, was removed and placed at room temperature for 30 minutes, the CTG reagent was also removed and equilibrated to room temperature, 15 μl of CTG reagent was added to each well, and placed on a shaker with gentle shaking for 5 minutes to ensure adequate cell lysis, placed for 10 minutes to stabilize the luminescence signal, and then EnVision (Perkin Elmer) was used to read the luminescence signal. In addition, for correcting the cell number, a T ₀ control is simultaneously arranged, wherein the T ₀ control comprises a blank control only containing a culture medium and a cell adding control, the difference value of the blank control and the cell adding control is set as a T ₀ control, and the cell adding control is obtained by adding a CTG reagent before adding medicine.

The percentage of inhibition of LC-2/ad cell proliferation by a compound can be calculated by the following formula:

percent inhibition =100-100*{[(signal_{Compounds of formula (I)}-Signal_{Negative control})-T_{0 control}]/[(signal_{positive control}-Signal_{Negative control})-T_{0 control}]}.

Y = Bottom + (Top- Bottom)/(1+10^((LogIC₅₀-X) * slope factor))

Determination of TT cell half-effective inhibition concentration GI ₅₀

The effect of the compounds of the invention on TT cell proliferation was evaluated using a luminescent cell viability test experiment.

The experimental procedure is summarized as follows:

using CELLTILTER-Glo (CTG) assay kit, cell proliferation status of TT was detected by detecting an indicator ATP of viable cell metabolism using a unique, stable luciferase, the luminescence signal generated in the assay being proportional to the number of viable cells in the medium.

TT cells (ATCC, CRL-1803) were cultured in F12K medium (Thermofisher, 21127022) containing 10% FBS (GBICO, 10099-141) and 100units/ml of a mixture of green streptomycin (Thermofisher, 15140122), and the cultured cells were digested and blown off with 0.25% pancreatin (Thermofisher, 25200056) and then plated in white 384-well plates (Thermofisher, 164610) with 27. Mu.L of the medium containing the cells per well, and then the 384-well plates were placed in an incubator containing 5% CO ₂ at 37℃overnight. Compounds were dissolved in 100% DMSO and diluted to 1mM, after which 4-fold serial dilutions were made in DMSO to a minimum concentration of 0.061. Mu.M, and each concentration point was diluted 50-fold with F12K medium. If the compound GI ₅₀ value is very low, the initial concentration of the compound can be reduced. mu.L of diluted compound was added to each well and gently centrifuged to mix. Wherein, the culture medium without cells was used as a negative control (100% inhibition), and a 0.2% DMSO group was used as a positive control (0% inhibition). The 384 well plate was placed in an incubator at 37℃with 5% CO ₂ for continuous incubation, after 96 hours, was removed and placed at room temperature for 30 minutes, the CTG reagent was also removed and equilibrated to room temperature, 30. Mu.L of CTG reagent was added to each well, placed on a shaker with gentle shaking to ensure adequate cell lysis, placed for 10 minutes to stabilize the luminescence signal, and then EnVision (Perkin Elmer) was used to read the luminescence signal. In addition, for correcting the cell number, a T ₀ control is simultaneously arranged, wherein the T ₀ control comprises a blank control only containing a culture medium and a cell adding control, the difference value of the blank control and the cell adding control is set as a T ₀ control, and the cell adding control is obtained by adding a CTG reagent before adding medicine.

The percent inhibition of TT cell proliferation by a compound can be calculated using the following formula:

percent inhibition =100-100*{[(signal_{Compounds of formula (I)}-Signal_{Negative control})-T_{0 control}]/[(signal_{positive control}-Signal_{Negative control})_-T_{0 control}]}

Y = Bottom + (Top- Bottom)/(1+10^((LogIC₅₀-X) * slope factor))

The results of the in vitro kinase assay described above are shown in Table 1 below and the results of the cell assay are shown in Table 2 below.

Table 1: in vitro kinase Activity assay results

Numbering of compounds	RET IC₅₀(nM)	RET V804M IC₅₀ (nM)	RET V804L IC₅₀ (nM)	RET M918T IC₅₀ (nM)	VEGFR2 IC₅₀ (nM)
						23	0.95	0.47	1.07
44	0.88
						142	2.47	0.99	1.31
143	0.21	0.48		3.52	44.19
						144	0.18		0.59	40.24
145	0.38			5.41	71.86
						146	0.26		1.01
147	0.26	0.31		0.89	10.83
						148	1		6.94	36.67
149	1.66			6.27	64.56
						150	1.67		8.69	30.47
151	3.38
						152	3.23	3.08	7.76	30.81
153	0.27	0.23		0.52	21.70
						154	10.63
155	2.32	1.93		2.87	14.55
						156	0.98		2.92	10.9
157	2.36	3.96		4.63	6.5
						158	34.65
159	2.16	1.39		6.96	6.82
						160	2.02	2.19	5.45	17.29
161	1.71	0.23		3.21	125.96
						162	0.06	0.05	0.66	7.58
163	1.56
						164	0.62
165	69.84
						166	79.86
167	25.18
						168	0.47		1.32
169	1.13	0.29		1.30	292.72
						170	1.18	1.42
171	0.67	0.63		7.13	340.12
						172	1.37
173	0.99			5.77
						174	3.4
175	0.29	0.28		1.34	221.57
						176	3.73	1.38	14.21	113.33
177	3.74	1.05		17.94	38.86
						178	19.11
179	0.29	0.06		1.84	206.82
						180	0.9		4.75	413.35
181	4.41
						183	0.24	0.11	2.53	41.63
184	6.59
						185	1.18		2.46	81.63
186	3.94	1.59		9.63	201.11
						187	0.89	0.51	3.49	75.19
188	1.93	0.44		3.80	45.91
						189	4.79
190	3.47
						191	13.53
192	0.18	0.21		1.96	84.52
						193	0.35	0.34	3.55	86.83
194	1.32	1.81		9.84	53.95
						195	0.31	0.14	1.56	48.19
196	5.31
						197	4.72
198	1.48	1.22		9.98	80.83
						199	2.31
200	15.08
						201	1.96	1.85	10.9	79.94
202	4.68	2.31		25.51	169.82
						203	1.38	2.55	29.81	67.9
205	1.97			4.79
						206	8.91		14.55
207	3.47
						208	8.5
209	8.2
						210	3.84
211	1.53
						212	78.04
213	25.34
						214	3.63		12.15
215	10.82
						216	2.82	0.43	4.33	93.79
217	8.76
						218	7.83		39.83	194
219	9.55			23.1	119.28
						220	12.6		34.62	160.37
221	14.37			48.34	81.68
						222	6.01		22.57	144.73
223	4.92			46.76	95.63
						224	4.47		22.96	104.97
225	13.9
						226	8.72
227	30.7
						228	2.88		16.15	54.91
229	11.42
						230	5.85		12.62	36.94
231	9.24			28.22	33.17
						232	16.21
233	17.55
						234	8.57		22.25	68.82
235	0.35			20.93	32.31
						236	11.77
237	11.83
						238	23.18
239	38.27
						240	10.37
241	26.39
						242	7.59			67.38
243	6.2				29.5
						244	25.18
245	4.88			20.88	43.76
						246	2.39			38.47
247	2.15			8.77	33.56
						248	3.75		18.31	53.24
249	13.07
						250	24.42
251	15.12
						252	6.61		64.47	32.24
253	16.86			59.7	47.96
						254	22.91
255	13.34			62.72	44.71
						256	26.84
257	15.45
						258	6.46		50.75	127.21
259	4.12			12.83	44.78
						260	3.61			29.61
261	28.37
						262	26.39
263	9.91
						264	11.95
265	30.44
						266	33.69
267	30.7
						268	4.63
269	22.42
						270	16.67
271	9.76
						272	14.02
273	30.14
						274	14.62
275	13.77
						276	74.39
277	30.79
						278	53.71
279	55.45
						280	26.39
281	13.34			62.72	44.71
						282	15.91		44.61	107.3
283	13.04			39.1	219.35
						284	12.09
285	17.69
						286	2.04		5.38	70.94
287	5.65			30.34	70.99
						288	6.58		10.78	111.16
289	11.46			17.11	129.67
						290	14.58		21.03	47.94
291	1.98			20.44	31.39
						292	0.8
293	10.74
						294	0.38		2.81	19.67
295	1.09			12.32	42.78
						296	3.59		16.33	44.31
297	3.47			10.49	55.78
						298	5.54		10.46	60.58
299	5.24			15.23	74.61
						300	9.27		12.22	53.3
301	2.98			5.7	46.08
						302	8.62		8.01	64.8
303	0.83			4.04	24.38
						304	1.04		8.54	54.33
305	3.88			39.55	61.38
						306	11.78
307	8.92			21.02	131.75
						308	17.57
309	17.78
						310	18.29
311	23.75
						312	10.64
313	21.22
						314	16.86
315	9.39			22.97	129.8
						316	26
317	15.12
						318	21.47
319	3.01			79.11	460.54
						320	26.92		19.42
321	4.57
						322	8.71
323	7.69			13.8	36.55
						324	9.24		17.91	21.84
325	3.06
						326	10.08
327	2.91			10.51	28.79
						328	3.22		5.2	61.96
329	7.05				47.67
						330	2.34		6.06
331	1.64			4.47
						332	0.82		1.31
333	10.25
						334	3.27		9.45
335	1.27			20.48	899.16
						336	30.52
337	1.47			16.26	966.39
						338	1.59		12.58	679.27
339	23.9
						340	10.57
341	7.1				497.75
						342	64.14
343	59.22
						344	64.02
345	2.04			5.38	70.94
						346	54.97
347	7.04			20.09	50.67
						348	11.99
349	9.13			47.81	55.66
						350	16.55
351	22.74
						352	0.81		1.98	21.22
353	2.51
						354	3.96			157.23
355	3.04				196.25
						356	1.14			105.32
357	37.56
						358	23.66
359	20.44
						360	3.1		4.23
361	2.49			3.65
						362	1.2		1.23
363	14.15
						364	13.74
365	3.14			11.83
						366	5.44
367	19.02
						368	8.23		49.17	39.05
369	18.42			32.24	50.3
						370	12.27
371	26.39
						372	0.41		4.48	332.31
373	6.53			17.64	268.53
						374	1.53		3.02	179.11
375	2.22			4.62	132.69
						376	0.57		0.63	32.23
377	0.54			1.19	65.8
						378	2.67
379	5.07
						380	0.96		1.39	37.89
381	0.84			0.91	39.75
						382			13.06	349.93
383	4.76			7.95	90.59
						384	0.58		0.52	26.75
385	1.2			1.28	45.02
						386	0.86		0.81	78.61
387	1.61			1.22	48.39
						388	17.85		21.09	221.82
389	1.09			0.54	40.93
						390	2.6
391	6
						392	0.42		0.69	21.81
393	8.7
						394	0.87		1.13	142.77
395	2.52			2.37	99.44
						396	1.05		9.86	44.03
397	1.82			2.95	32.87
						398	0.56		0.59	45.47
399	0.63			0.94	37.58
						400	3.11		4.16	51.76
401	1.7			6.25	92.39
						402	1.46		3.39	58.8
403	0.77			1.68	78.19
						404	2.30		19.0	182.05
405	0.85			2.15	67.61
						406	1.42		6.65	58.84
407	9.16
						408	1.12		4.78	97.68
409	1.64			2.17	58.61
						410	1.04		1.66	46.13
411	2.1			2.89	63.34
						412	2.89		5.51	88.27
446	9.69			53.48	50.92
						447			7.09	60.54
448	6.06			13.38	211.38
						449	2		9.46	49.15
450	4.88			6.41	19.51
						451	7.28
461	2.66			13.58	36.67

Table 2: in vitro cell Activity detection results

Numbering of compounds	Ba/F3_KIF5B-RET GI₅₀ (nM)	LC-2/ad CCDC6 GI₅₀ (nM)	TT C634W ATCC GI₅₀ (nM)
				23	518.26	18.69
142	24.78	5.02	7.97
				143	244.26	12.08	5.36
144	27.94	8.09	4.52
				145	702.15	10.43	9.31
146	231.74	21.04
				147	105.35	2.42	1.25
148	965.61	32.23	59.35
				149		37.03	35.95
150	855.75	32.05	82.99
				151	559.84	59.45
152	211.93	18.48	19.97
				153	158.65	5.0	6.1
154	321.79	48.01
				155	136.42	37.28	18.59
156	67.51	25.07	7.75
				157	312.54	30.85	27.89
158		556.06
				159	135.49	22.36	18.39
160	173.65	17.9	16.84
				161	324.77	29.84	14.26
162	103.51	1.55	0.98
				164	720.31	21.1	19.54
165	1555.84
				168		66.96
169	197.39	10.14	2.31
				170	522.1	24.82	25.61
171	516.12	28.99	25.41
				172		28.02
173	516.84	12.44
				174	318.77	31.31
175	334.26	7.98	5.88
				176	274.71	36.29	45.23
177	268.76	28.28	51.6
				179	260.85	7.51	5.33
180	396.60	18.43
				181	883.77	66.68
183	157.62	8.83	5.04
				184	237.32	36.75
185	136.02	13.41	7.67
				186	229.57	34.43	12.22
187	241.42	14.79	9.83
				188	192.05	15.42	19.3
189	362.33
				190	321.38	37
192	210.86	3.85	3.91
				193	147.57	4.45	7.43
194	271.67	13.02	31.22
				195	215.15	9.44	3.54
198	243.31	19.21	27.58
				199	318.54	41.3
201	277.11	17.52	42.56
				202	187.4	27.64	47.68
203	215.55	25.4	38.01
				205		42.8
206	898.33	70.89
				207		103.63
210		96.36
				211		68.86
213		178.56
				214	274.61	92.4
215	601.01	230.46
				216	202.1	40.62	15.34
218	971.55
				223	793.65
228	397.3
				230	473.18	102.92	249.12
231	615.5	126.95	301.75
				234	491.32	63.8	151.26
235	326.84	85.93	208.72
				237		203.39	584.34
242	888.52	198.36	469.38
				243	903.01	158.22	214.51
245	386.43	93.42	233
				246	865.63	146.35
247	799.02	469.39	878.81
				248	578.42	403.94	453.02
249		406.54	855.56
				252	343	258.82	426.81
253	887.99	401.30	791.34
				255	617.08	362.79	700.30
259	638.64	44.61	135.88
				260	871.38	229.90	369.14
271	968.26
				275		234.94	548.45
281	617.08	362.79	700.30
				286	330.94	34.45	84.8
287	857.68	194.76	486.51
				288	756.40	320.47	530.4
289	445.16	348.23	492.58
				290	716.69	500.41	706.35
291	459.59	79.19	179.69
				292		71.72	187.11
293		688.03
				294	277.87	136.85	176.79
295	589.70	45.39	328.88
				296	417.86	360.56	297.38
297	494.17	354.23	252.55
				298	415.28	354.77	207.53
299	436.13	315.01	450.7
				300	323.69	330.95	349.79
301	377.79	294.01	227.15
				302	713.96
303	422.3	354.78	362.22
				304	348.09	316.17	277.7
305	991.95	262.44	430.52
				308	774.67
309	903.56
				310	598.31
311	656.03
				312	794.56
313	646.35
				314	583.37
315	651.85
				316	708.98
317	216	298.86	438.62
				318	514.63
320	467.99	488.48	778.82
				322		344.87	732.16
323		378.61	123.46
				324	746.25	290.41	425.1
327	493.06	69.78	171.7
				329	933.23
332	563.19
				345	330.94	34.45	84.8
347	285.11	63.74	136.98
				349	614.76	191.09	330.19
352	619.78	99.14	425.4
				353	677.96
354	939.15
				356	942.56
362	879.96	144.85	127.41
				366		146.98	143.05
368	745.78	149.3	296.28
				369	827.98	343.55	721.41
372	650.16
				374	748.49
376	464.22	88.88	44.03
				377	385.11	139.27	118.43
380	622.27		115.24
				381	787.76
383	831.02
				384	772.76
386		228.50	150.29
				387	657.42	134.61	134.56
388
				389	693.6	100.44	62.95
390	863.65
				392	607.20	78.54	58.81
394		120.26	51.71
				395		203.92	96.59
396	512.03	94.22	65.41
				397		243.47	151.24
398	736.92	52.86	27.56
				399	636.95	91.1	38.26
400		467.94	517.93
				401		333.91	281.8
402		311.63	278.06
				403		145.9	154.65
404	811.76	188.48	203.78
				405	641.09	77.65	58.63
406	717.75	135.01	173.48
				407	828.47		244.87
408	685.15	96.42	125.23
				409	506.99	94.44	79.31
410	445.39	66.25	61.38
				411	499.71	119.39	100.08
412	417.64	123.33	109.82
				446	314.32	84.74	237.43
447	220.16	217.22	421.58
				448	473.19	201.59	283.41
449	439.24	468.31	315.1
				450	328.53	425.29	470.94
451		840.01	853.51
				461	466.35	57.4	159.11

From the above experimental results, it is known that the compounds of the embodiments of the present invention can effectively inhibit the activity of RET kinase, and can be used for treating RET kinase mediated diseases, such as cancers, especially hematological malignancies, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, and glioma. Some compounds can also effectively inhibit proliferation of Ba/F3_KIF5B-RET tumor cells, LC-2/ad tumor cells and TT tumor cells.

It will be apparent to those skilled in the art that the present disclosure is not limited to the illustrative embodiments described above, and that it may be embodied in other specific forms without departing from the essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, reference being made to the appended claims rather than to the foregoing embodiments, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

Claims

1. A compound according to formula I or a pharmaceutically acceptable salt thereof:

Wherein:

x ¹ is CH or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

y ¹ is CH or N;

Y ² is CH or N;

y ³ is CH or N;

y ⁴ is CH or N;

And one of the following conditions is satisfied: y ¹、Y²、Y³ and Y ⁴ are both CH; only Y ¹ or Y ² is N; or Y ¹ and Y ³ are both N; a is hydrogen;

b is selected from hydrogen, halogen, -OR ¹⁴ OR

d is selected from:

And when X ² is CH, D is not

Ar ² is selected from 5-6 membered aryl or heteroaryl optionally mono-or di-substituted, said substituents being selected from halogen or C1-C6 alkoxy; r ^m、Rⁿ is each independently selected from hydrogen, hydroxy, C1-C6 alkyl or hydroxy C1-C6 alkyl, or R ^m、Rⁿ together with the carbon atom to which it is attached form C3-C6 cycloalkyl;

r is 0, 1,2 or 3.

2. A compound of formula I as defined in claim 1, or a pharmaceutically acceptable salt thereof, wherein

X ¹ is CH or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

a is hydrogen;

b is selected from hydrogen, halogen, -OR ¹⁴ OR

d is selected from:

And when X ² is CH, D is not

r is 0, 1 or 2.

3. A compound of formula I as defined in claim 1, or a pharmaceutically acceptable salt thereof, wherein

X ¹ is CH or N;

X ² is CH or N;

Provided that up to 1 of X ¹、X² is N;

a is hydrogen;

b is selected from hydrogen, halogen, -OR ¹⁴ OR

R ¹⁵ is C1-C4 alkyl;

D is selected from

And when X ² is CH, D is not

When D is When E is selected from amino, -NHC (O) R ^X or-SO ₂ (C1-C4) alkyl;

When D is When E is pyridine-2-oxy;

When D is When E is selected from the group consisting of-C (O) R ^y,-SO ₂ (C1-C4) alkyl, or-CH ₂-Ar²;

r is 0, 1 or 2.

4. A compound of formula I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is

5. Use of a compound of formula I according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for use as a RET kinase inhibitor.

6. Use of a compound of formula I according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment or prophylaxis of a related disorder mediated by RET kinase, said related disorder being a tumor selected from the group consisting of hematological malignancies, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, brain glioma.

7. A pharmaceutical composition comprising a compound of formula I according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, optionally one or more other RET kinase inhibitors, and one or more pharmaceutically acceptable carriers, diluents and excipients.