WO2020088390A1 - Pyrimidopyrazole compounds as fourth generation egfr inhibitors - Google Patents

Abstract

Disclosed are novel pyrimidopyrazole compounds as fourth generation (T790M/C797S mutation) EGFR kinase inhibitors and pharmaceutical use thereof, specifically disclosed are a compound represented by formula (I), a stereoisomer, a racemate, a tautomer, an isotope label, a nitrogen oxide or a pharmaceutically acceptable salt thereof. Said compounds have good efficacy in treating diseases caused by abnormal mutation of EGFR Del19/T790M/C797S and L858R/T790M/C797S.

Description

Pyrimidine pyrazoles as fourth-generation EGFR inhibitors

This application requires that it be submitted to the State Intellectual Property Office of China on October 29, 2018. The patent application number is 201811266972.X, and the name of the invention is "pyrimidine pyrazole compounds as fourth-generation EGFR inhibitors" Submitted by the State Intellectual Property Office of China, the patent application number is 201910479288.8, and the priority of the two prior applications is the invention titled "Pyrimidine pyrazoles as fourth-generation EGFR inhibitors." The full texts of the two applications are incorporated by reference in this application.

Technical field

The present invention relates to novel pyrimidine pyrazole compounds as fourth-generation (T790M / C797S mutation) EGFR kinase inhibitors and their medical applications, and specifically discloses compounds represented by formula (I), their stereoisomers, racemates, Tautomers, isotopic labels, nitrogen oxides or pharmaceutically acceptable salts thereof.

Background technique

EGFR (Epidermal Growth Factor Receptor) is a receptor for epithelial growth factor (EGF) cell proliferation and signaling. EGFR belongs to a family of ErbB receptors, which includes EGFR (ErbB-1), HER2 / c-neu (ErbB-2), HER3 (ErbB-3) and HER4 (ErbB-4). EGFR is also called HER1, ErbB1. EGFR is widely distributed on the cell surface of mammalian epithelial cells, fibroblasts, glial cells, keratinocytes, etc. EGFR signaling pathway plays an important role in the physiological processes of cell growth, proliferation and differentiation. EGFR is divided into three regions: extracellular ligand binding region, transmembrane region and intracellular kinase region. When the EGFR is bound by the corresponding ligand, it will induce the formation of homo- or heterodimers, thereby activating the intracellular tyrosine kinase pathway to phosphorylate itself, thereby guiding downstream phosphorylation, including MAPK, Akt and JNK Pathway to induce cell proliferation.

EGFR tyrosine kinase inhibitor (Tyrosine Kinase Inhibitor, TKI) is to block endogenous ATP binding to the intracellular kinase region, thereby inhibiting receptor phosphorylation and the activation of downstream signal transduction molecules, blocking tumors Cell proliferation. Although EGFR targeted therapy has successfully entered the clinical stage and drugs have been marketed, EGFR gene mutations have led to drug resistance. The mutations mainly occur in exons 18-21, of which the deletion of exon 19 and the point mutation of L858R in exon 21 are the most common mutation subtypes, accounting for 90% of all mutation types. With the development and use of drugs, most drug resistance appears in the second mutation of T790M in the gatekeeper region of kinases. The three generations of irreversible inhibitors developed in recent years have good inhibitory activity against the T790M mutation, but they also inevitably have the C797S mutation, and are the main drug resistance mechanism (about 40%) of the star drug AZD9291. In 2018, AZD9291 has entered the first-line treatment. In response to the C797S mutation, there is an urgent need to develop a new, safer and more effective EGFR TKI.

Summary of the invention

In order to solve the drug resistance problem of the third-generation inhibitors, that is, the third mutation C797S problem of EGFR, the present invention provides the compound represented by formula (I), its stereoisomers, racemates, and Variant isomers, isotope labels, nitrogen oxides or pharmaceutically acceptable salts thereof,

Where E is selected from

Y is selected from O or S; R ₁ , R ₂ , R ₃ , R _{4 are the} same or different, and are independently selected from C _1-12 alkyl;

A is selected from N or C-Q;

B is selected from N or C-D;

Q and D are the same or different and are independently selected from H, halogen, cyano, hydroxy, amino, unsubstituted or optionally substituted with one, two or more R _a of the following groups: C _1-12 alkane group, C _1-12 heteroalkyl, C ₂ - ₁₂ alkenyl group, C _2-12 alkenyl, heteroaryl, C ₂ - ₁₂ alkynyl group, C _2-12 heteroaryl alkynyl, C _1-12 alkoxy, C _{3 --20} cycloalkyl, 3-20 membered heterocyclyl, C ₆ - ₂₀ aryl, 5-20 membered heteroaryl, -COOC _1-12 alkyl, -COC _1-12 alkyl;

Each R _{a is the} same or different and is independently selected from halogen, cyano, amino, hydroxy, unsubstituted or optionally substituted with one, two or more Rs of the following groups: C _1-12 alkyl, C _1-12 heteroalkyl, C _1-12 alkoxy, C ₃ - ₂₀ cycloalkyl, 3-20 membered heterocyclyl, -COOC _1-12 alkyl, -COC _1-12 alkyl;

V and Z are the same or different and are independently selected from H, halogen, cyano, hydroxy, amino, unsubstituted or optionally substituted with one, two or more R _b of the following groups: C _1-12 alkane group, C _1-12 heteroalkyl, C ₂ - ₁₂ alkenyl group, C _2-12 alkenyl, heteroaryl, C ₂ - ₁₂ alkynyl group, C _2-12 heteroaryl alkynyl, C _1-12 alkoxy, C _{3 --20} cycloalkyl, 3-20 membered heterocyclyl, C ₆ - ₂₀ aryl, 5-20 membered heteroaryl, -COOC _1-12 alkyl, -COC _1-12 alkyl, -N (C _{1 -12} alkyl) ₂ , -NHC _1-12 alkyl;

Each R _{b is the} same or different and is independently selected from halogen, cyano, hydroxy, amino, unsubstituted or optionally substituted with one, two or more Rs of the following groups: C _1-12 alkyl, C _1-12 alkoxy, C _1-12 heteroalkyl, C ₃ - ₂₀ cycloalkyl, 3-20 membered heterocyclyl;

Each Rs is the same or different and is independently selected from halogen, CN, OH, C _1-12 alkyl;

W is selected from the group consisting of hydrogen, unsubstituted or optionally substituted with one, two or more R _c : C _1-12 alkyl, C _1-12 heteroalkyl, C _1-12 alkanoyl, C _1-12 alkanoyl heteroaryl, 3-20 membered heterocyclyl group, C ₃ - ₂₀ cycloalkyl, 3-20 membered heterocyclyl, C ₆ - ₁₈ aryl, 5-20 membered heteroaryl, C ₃ - ₂₀ cycloalkyl and C ₃ - ₂₀ cycloalkyl group formed spiro cycloalkyl group, C ₃ - ₂₀ cycloalkyl group spiro ring and 3-20 membered heterocyclic group formed, 3-20 membered heterocyclic group 3-20 membered Spirocyclic group formed by heterocyclic group, C3-20 cycloalkyl and C3-20 cycloalkyl cyclic group, C3-20 cycloalkyl and 3-20 membered heterocyclic group Cyclocyclic group, 3-20 member heterocyclic group with 3-20 membered heterocyclic group and cycloalkyl group, C ₅ - ₁₄ bridged cycloalkyl groups, 5-14 membered bridged ring heteroaryl group;

Each R _{c is the} same or different, and is independently selected from the group consisting of = O, halogen, cyano, hydroxy, amino, carboxy, unsubstituted or optionally substituted with one, two or more _Rd : C _1-12 alkyl, C _1-12 alkoxy, C _1-12 heteroalkyl, -S (O) ₂ C _1-12 alkyl, -C (O) ₂ C _1-12 alkyl, -COC ₁₂ alkyl, C ₃ - ₂₀ cycloalkyl, 3-20 membered heterocyclyl;

Each _{Rd is the} same or different, and is independently selected from halogen, = O, halogen, cyano, hydroxyl, -S (O) ₂ C _1-12 alkyl, -COOC _1-12 alkyl, -COC _{1- 12} alkyl group, C _1-12 alkyl group; when W is selected from 3-20 membered heterocyclyl, C ₃ - ₂₀ cycloalkyl group spiro ring and 3-20 membered heterocyclic group formed, 3-20 membered heterocyclyl When the group is a spiro ring group formed from a 3-20 membered heterocyclic group and a 5-14 membered heterobridged ring group, the substituted R _c may be connected by a hetero atom or a carbon atom.

According to an embodiment of the present invention, Q and D are identical or different and independently selected from H, halo, cyano, hydroxy, amino, unsubstituted or optionally substituted with one, two or more of the following R _a substituents group Group: C _1-6 alkyl, C _1-6 heteroalkyl, C _1-6 alkenyl, C _1-6 heteroalkenyl, C _1-6 alkynyl, C _1-6 heteroalkynyl, C _{3- 6} cycloalkyl, C _3-6 heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

V and Z are the same or different and are independently selected from H, halogen, cyano, hydroxy, amino, unsubstituted or optionally substituted with one, two or more R _b of the following groups: C _1-6 alkane Group, C _1-6 heteroalkyl, C _3-6 cycloalkyl, C _3-6 heterocycloalkyl, phenyl and 5-6 membered heteroaryl, the C _1-6 alkyl, C _{1 -6} heteroalkyl, C _3-6 cycloalkyl, C _3-6 heterocycloalkyl, phenyl and 5-6 membered heteroaryl;

W is selected from the group consisting of hydrogen, unsubstituted or optionally substituted with one, two or more R _c : C _1-6 alkyl, C _1-6 heteroalkyl, C _1-3 alkanoyl, C _1-3 Heteroalkanoyl, C _3-8 cycloalkyl, C _3-8 heterocyclyl, C _3-6 cycloalkyl and C _3-6 cycloalkyl spiro, C _3-6 cycloalkyl Spirocyclic group with C _3-6 heterocyclic group, C _3-6 cycloalkyl group and C _3-6 cycloalkyl group, C _3-6 cycloalkyl group and C _3-6 heterocyclic group Cyclocyclic group, C _3-6 heterocyclic group and C _3-6 heterocyclic group, C _5-14 bridge ring group, C _5-14 hetero bridge ring group;

According to an embodiment of the present invention, each R _a, R _b, R _c identical or different, each independently selected from -F, -Cl, -Br, -I, -OH, -CN, -NH 2, -CH ₃ , CH ₃ CH _2- , -CF ₃ , -CHF ₂ , -CH ₂ F, NH ₂ CH _2- , -NH (CH ₃ ) ₂ , -OCH ₃ , CH ₃ CH ₂ O-, CH ₃ OCH ₂ -, MeSO _2- , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ , -CH ₂ C (CH ₃ ) ₂ OH, -CH ₂ CH ₂ SO ₂ Me, -CH ₂ OH, -CH ₂ CH ₂ OH , -CH ₂ CH ₂ CN, -CH ₂ CH ₂ OCH ₃ , -COOCH (CF ₃ ) ₂ , -COCH ₂ OH, -CH ₂ COOH, -CH ₂ CONH ₂ , 2- (1-morpholinyl) ethyl Group, 1-methylpiperidin-4-acyl, 3-pyridyl, 2- (pyrrolidin-1-yl) ethyl, 3-oxetanyl, tetrahydropyrrolyl, morpholinyl, 4- Methyl-1-piperazinyl, 1-methyl-4-piperidinyl;

According to an embodiment of the present invention, each Rs is the same or different and is independently selected from halogen, CN, OH, C _1-6 alkyl;

Preferably, E is selected from

Y is selected from O or S; R ₁ , R ₂ , R ₃ , and R _{4 are the} same or different, and are independently selected from C _1-6 alkyl; A is selected from N or CQ; B is selected from N or CD; A and At least one of B is N, and A and B are not N at the same time;

Q and D are the same or different and are independently selected from H, halogen, cyano, C _1-6 alkyl, halogen substituted C _1-6 alkyl, C _1-6 heteroalkyl, C _1-6 alkoxy group, C ₃ - ₁₂ cycloalkyl, 3-12 membered heterocyclyl, -COOC _1-6 alkyl, -COC _1-6 alkyl;

V and Z identical or different, independently selected from H, halo, cyano, C _1-6 alkyl, C _1-6 alkoxy, C ₃ - ₆ cycloalkyl;

W is selected from unsubstituted or optionally substituted by one, two or more R _c is the group: C _1-6 alkyl, C _1-6 heteroalkyl, C ₃ - ₁₂ cycloalkyl group, 3- 12-membered heterocyclic, C ₆ - ₁₂ aryl, 5-12 membered heteroaryl, C ₃ - ₆ cycloalkyl and C ₃ - ₆ cycloalkyl group formed spiro cycloalkyl group, C ₃ - ₁₂ cycloalkyl and The spirocyclic group formed by the 3-12 membered heterocyclic group, the spirocyclic group formed by the 3-12 membered heterocyclic group and the 3-12 membered heterocyclic group, the combination of the C _3-6 cycloalkyl group and the C3-6 heterocyclic group cycloalkyl group, C3-6 heterocyclyl and C3-6 cycloalkyl and heterocyclyl group, C ₅ - ₁₄ bridged cycloalkyl groups, 5-14 membered bridged ring heteroaryl group;

According to an embodiment of the present invention, each R _a and R _{b are the} same or different and are independently selected from each other. R is selected from -F, -Cl, -Br, -I, -OH, -CN, -NH ₂ , and -CH ₃ , CH ₃ CH _2- , -CF ₃ , -CHF ₂ , -CH ₂ F, NH ₂ CH _2- , -NH (CH ₃ ) ₂ , -OCH ₃ , CH ₃ CH ₂ O-, CH ₃ OCH ₂ -, MeSO _2- , -CH ₂ CH ₂ F, -CH ₂ CHF ₂ , -CH ₂ C (CH ₃ ) ₂ OH, -CH ₂ CH ₂ SO ₂ Me, -CH ₂ OH, -CH ₂ CH ₂ OH , -CH ₂ CH ₂ CN, -CH ₂ CH ₂ OCH ₃ , -COOCH (CF ₃ ) ₂ , -COCH ₂ OH, -CH ₂ COOH, 1-tetrahydropyrrolyl, 2- (1-morpholinyl) Ethyl, 1-methylpiperidin-4-acyl, 3-pyridyl, 2- (pyrrolidin-1-yl) ethyl;

More preferably, E is selected from

Y is selected from O or S;

At least one of A and B is N, and A and B are not N at the same time;

Q and D are the same or different, and are independently selected from H, F, Cl, Br, trifluoromethyl, cyano, isopropyl, methoxy, cyclopropyl, -COOC ₂ H ₅ ;

V and Z are the same or different, and are independently selected from H, F, Cl, Br, methyl, methoxy, cyclopropyl;

W is selected from: tert-butyl,

In some preferred embodiments, when W is

Z is methyl, V is H, B is N, and E is

When Y is O, A is CD, D is selected from: halogen, C1-6 alkoxy, C3-6 cycloalkyl; when W is

Z is methoxy, V is H, B is N, and E is

When Y is O, A is CD, D is selected from: C1-6 alkyl, halogen substituted C1-6 alkyl, C1-6 alkoxy, C3-6 cycloalkyl, C1-6 heteroalkyl; W is

Z is methyl, V is H, A is N, and E is

When Y is O, B is CQ, and Q is selected from halogen, C1-6 alkyl, halogen-substituted C1-6 alkyl, C1-6 alkoxy, C3-6 cycloalkyl, and C1-6 heteroalkyl.

More preferably, the compound of formula (I) is selected from the following compounds or pharmaceutically acceptable salts thereof:

Wherein, the salt is hydrochloride, trifluoroacetate or formate.

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the compound represented by formula (I), its stereoisomer, racemate, tautomer, isotope label, nitrogen oxide or At least one of pharmaceutically acceptable salts.

Preferably, the pharmaceutical composition further includes a pharmaceutically acceptable excipient.

The present invention also provides at least one of the compound represented by formula (I), its stereoisomer, racemate, tautomer, isotopic label, nitrogen oxide, or pharmaceutically acceptable salt thereof. Use in the preparation of drugs for the treatment of cancer. Preferably, the cancer is non-small cell lung cancer, glioblastoma, pancreatic cancer, head and neck cancer, breast cancer, colorectal cancer, epithelial cancer, ovarian cancer, prostate cancer, adenocarcinoma or squamous cell carcinoma.

The present invention also provides at least one of the compound represented by formula (I), its stereoisomer, racemate, tautomer, isotopic label, nitrogen oxide, or a pharmaceutically acceptable salt thereof Application in preparing medicine for treating diseases caused by EGFR mutation.

Preferably, the EGFR mutation is one, two or more mutations selected from: (1) Del19; (2) T790M; (3) C797S; (4) L858R; (5) T790M; (6) C797S .

The present invention also provides a method for treating cancer, comprising: combining a compound represented by formula (I), its stereoisomer, racemate, tautomer, isotope label, nitrogen oxide, or its pharmaceutical At least one of the acceptable salts is administered to individuals in need.

Beneficial effect: The results of in vitro activity experiments prove that the compound of the present invention has a good potential therapeutic effect on diseases caused by abnormal mutations of EGFR Del19 / T790M / C797S and L858R / T790M / C797S.

Definition and description of terms

In the present invention

The position indicates the connection site.

Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A specific term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding trade product or its active ingredient.

Unless otherwise stated, the definitions of groups and terms described in the specification and claims of this application include their definitions as examples, exemplary definitions, preferred definitions, definitions listed in tables, definitions of specific compounds in the examples Etc., can be arbitrarily combined and combined with each other. Such combination and combination of group definitions and compound structures should fall within the scope of the description of this application.

Among them, "more" means three or more.

The term "halogen" refers to F, Cl, Br and I. In other words, F, Cl, Br and I can be described as "halogen" in this specification.

The term "C _1-12 alkyl group" is understood to mean preferably a linear or branched saturated monovalent hydrocarbon group having 1 to 12 carbon atoms, preferably a C _1-10 alkyl group. "C _1-10 alkyl" is understood to preferably mean a linear or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl Group, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, 1,2-dimethylbutyl, etc. or their isomers. In particular, the group has 1, 2, 3, 4, 5, 6, carbon atoms ("C _1-6 alkyl"), such as methyl, ethyl, propyl, butyl, isopropyl , Isobutyl, sec-butyl, tert-butyl, more particularly, the group has 1, 2 or 3 carbon atoms ("C _1-3 alkyl"), such as methyl, ethyl, n-propyl Base or isopropyl.

The term "C _2-12 alkenyl" is understood to mean preferably a linear or branched monovalent hydrocarbon group which contains one or more double bonds and has 2 to 12 carbon atoms, preferably "C _2-10 alkenyl" , Further preferably, "C _2-6 alkenyl". "C _2-10 alkenyl" is understood to mean preferably a linear or branched monovalent hydrocarbon group, which contains one or more double bonds and has 2, 3, 4, 5, 6, 7, 8, 9 or 10 Carbon atoms, especially 2 or 3 carbon atoms ("C _2-3 alkenyl"), it should be understood that in the case where the alkenyl group contains more than one double bond, the double bonds may be separated from each other yoke. The alkenyl group is, for example, vinyl, allyl, (E) -2-methylvinyl, (Z) -2-methylvinyl, (E) -but-2-enyl, (Z)- But-2-enyl, (E) -but-1-enyl, (Z) -but-1-enyl, pent-4-enyl, (E) -pent-3-enyl, (Z) -Pent-3-enyl, (E) -pent-2-enyl, (Z) -pent-2-enyl, (E) -pent-1-enyl, (Z) -pent-1-ene Group, hex-5-enyl, (E) -hex-4-enyl, (Z) -hex-4-enyl, (E) -hex-3-enyl, (Z) -hex-3- Alkenyl, (E) -hex-2-enyl, (Z) -hex-2-enyl, (E) -hex-1-enyl, (Z) -hex-1-enyl, isopropenyl , 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E) -1-methylprop-1-enyl, ( Z) -1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methyl Butyl-2-enyl, (E) -2-methylbut-2-enyl, (Z) -2-methylbut-2-enyl, (E) -1-methylbut-2- Alkenyl, (Z) -1-methylbut-2-enyl, (E) -3-methylbut-1-enyl, (Z) -3-methylbut-1-enyl, (E ) -2-methylbut-1-enyl, (Z) -2-methylbut-1-enyl, (E) -1-methylbut-1-enyl, (Z) -1-methyl Butyl-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethyl Prop-1-enyl, 1-propyl vinyl, 1-isopropyl vinyl. Those skilled in the art can understand that the term "C _1-n alkenyl" (for example, "C _1-12 alkenyl" or "C _1-6 alkenyl") is equivalent to the above definition of "C _2-n alkenyl" (Eg C _2-12 alkenyl).

The term "C ₂ - ₁₂ alkynyl group" is understood to mean a linear or branched divalent hydrocarbon chain comprising one or more triple bonds and having 2 to 12 carbon atoms, preferably "C ₂ -C ₁₀ alkynyl" , Further preferably, "C ₂ -C ₆ alkynyl". The term "C ₂ -C ₁₀ alkynyl" is understood to mean preferably a linear or branched monovalent hydrocarbon group, which contains one or more triple bonds and has 2, 3, 4, 5, 6, 7, 8, 9 Or 10 carbon atoms, especially 2 or 3 carbon atoms ("C ₂ -C ₃ -alkynyl"). The alkynyl group is, for example, ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl , Pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hex-4-ynyl, Hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-3-ynyl, 1-methylbut-2-ynyl , 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpentane -4-ynyl, 2-methylpent-3-ynyl, 1-methylpent-3-ynyl, 4-methylpent-2-ynyl, 1-methylpent-2-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-ethylbut-3-ynyl, 1-ethylbut-3-ynyl, 1-ethylbut- 2-alkynyl, 1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl, 2,2-dimethylbut-3-ynyl, 1,1-dimethylbutanyl -3-ynyl, 1,1-dimethylbut-2-ynyl or 3,3-dimethylbut-1-ynyl. In particular, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl. Those skilled in the art will understand that the term "C _1-n alkynyl" (for example, "C _1-12 alkynyl" or "C _1-6 alkynyl") is equivalent to the above definition of "C _2-n alkynyl" (Eg C _2-12 alkynyl).

"Hetero" means a heteroatom or a heteroatom group, independently selected from -O-, -S-, = O, = S, -ON =, -C (= O) O-, -C (= O)-, -C (= S)-,-S (= O) 2-,-S (= O)-,-C (= O) NH-,-NH-,-C (= NH)-,-S (= O) 2NH-,-S (= O) NH- and -NHC (= O) NH-; -P-,-P (= O) Me2 any one of the above cases, the number of heteroatoms or heteroatom groups are Choose from 1, 2, or 3.

Those skilled in the art will understand that the definitions of the aforementioned terms “C _1-12 alkyl”, “C _2-12 alkenyl”, and “C _2-12 alkynyl” also apply to “C _1-12 heteroalkane group "," C _2-12 alkenyl heteroaryl "," C _2-12 alkynyl heteroaryl "relates to alkyl, alkenyl, alkynyl moieties.

The term "C _3-20 cycloalkyl" is understood to mean a saturated or unsaturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 20 carbon atoms, preferably "C _3-10 cycloalkyl". The term "C _3-10 cycloalkyl" is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. The C _3-10 cycloalkyl group may be a monocyclic hydrocarbon group, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or bicyclic Hydrocarbon groups such as decalin ring. The "C _3-20 cycloalkyl group" may further include, for example, a C _1-8 cycloalkyl group, a C _3-8 cycloalkyl group, a C _3-6 cycloalkyl group, or the like.

The term "3-20 membered heterocyclic group" means a saturated or unsaturated monovalent monocyclic or bicyclic hydrocarbon ring, which contains 1-5 heteroatoms independently selected from N, O, and S, preferably "3-10 membered" Heterocyclyl ". The term "3-10 membered heterocyclic group" means a saturated monovalent monocyclic or bicyclic hydrocarbon ring, which contains 1-5, preferably 1-3 heteroatoms selected from N, O, and S. The heterocyclic group may be connected to the rest of the molecule through any of the carbon atoms or nitrogen atom (if present). In particular, the heterocyclic group may include, but is not limited to: a 4-membered ring, such as azetidine, oxetanyl; a 5-membered ring, such as tetrahydrofuranyl, dioxolyl, pyrrole Alkyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or 6-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithialkyl, thiomorpholinyl, piperazinyl Or trithiane; or a 7-membered ring, such as diazacycloheptanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclic group may be bicyclic, such as but not limited to a 5,5 membered ring, such as hexahydrocyclopenta [c] pyrrole-2 (1H) -yl ring, or a 5,6 membered bicyclic ring, such as hexahydropyrrole And [1,2-a] pyrazine-2 (1H) -yl ring. The nitrogen-containing ring may be partially unsaturated, ie it may contain one or more double bonds, such as but not limited to 2,5-dihydro-1H-pyrrolyl, 4H- [1,3,4] thiadi Azinyl, 4,5-dihydrooxazolyl, or 4H- [1,4] thiazinyl, or it may be benzo-fused, such as but not limited to dihydroisoquinolinyl. According to the present invention, the heterocyclic group is non-aromatic.

The terms "C3-6 heterocycloalkyl" and "C3-6 heterocyclyl" correspond to a heterocyclic group having 3 to 6 C atoms, and are also covered by the above-mentioned "3-20 membered heterocyclic group".

The term "C _6-20 aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6 to 20 carbon atoms, preferably "C _6-14 aryl" . The term "C _6-14 aryl" is understood to preferably mean a monovalent aromatic or partially aromatic monocyclic, bicyclic or monocyclic aromatic ring having 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms Tricyclic hydrocarbon rings ("C _6-14 aryl"), especially those with 6 carbon atoms ("C ₆ aryl"), such as phenyl; or biphenyl, or those with 9 carbon atoms Ring ("C ₉ aryl"), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C ₁₀ aryl"), such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl, Either a ring with 13 carbon atoms ("C ₁₃ aryl"), such as fluorenyl, or a ring with 14 carbon atoms ("C ₁₄ aryl"), such as anthryl.

The term "5-20 membered heteroaryl" should be understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5 to 20 ring atoms and containing 1 to 5 independently selected from N, O And S heteroatoms, such as "5-14 membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, especially 5 or 6 or 9 or 10 carbon atoms, and it contains 1-5, preferably 1-3 heteroatoms independently selected from N, O and S and, in addition, in each case The bottom can be benzo fused. In particular, the heteroaryl group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thio Diazolyl, thi-4H-pyrazolyl, etc. and their benzo derivatives such as benzofuranyl, benzothienyl, benzoxazolyl, benzoisoxazolyl, benzimidazolyl, benzene Pyridazolyl, indazolyl, indolyl, isoindolyl, etc .; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., and their benzo derivatives, such as quinoline Group, quinazolinyl group, isoquinolinyl group, etc .; or azininyl, indazine group, purinyl group, etc. and their benzo derivatives; or cinnoline group, phthalazinyl group, quinazolinyl group, quinoxaline group Porphyrinyl, naphthyridyl, pteridyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, etc.

Unless otherwise stated, heterocyclyl, heteroaryl or heteroarylene includes all possible isomeric forms thereof, such as positional isomers. Thus, for some illustrative non-limiting examples, pyridyl or pyridinylene includes pyridin-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-3-yl, pyridin-4-yl And pyridin-4-yl; thienyl or thienylene includes thien-2-yl, thien-2-yl, thien-3-yl and thien-3-yl.

The term "C _5-14 bridged cyclic group" refers to a 5- to 14-membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected. These may contain one or more double bonds, but none of them The ring has a completely conjugated π electron system. Including 7 to 10 yuan. For example, the following groups:

According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridge cycloalkyl.

The term "C _{5-14 heterobridge} ring group" refers to a polycyclic group of 5 to 14 members, any two rings sharing two atoms that are not directly connected, these may contain one or more double bonds, but none of the rings has A completely conjugated π-electron system in which one or more ring atoms are selected from nitrogen, oxygen, or S (O) n (where n is an integer from 0 to 2) heteroatom, and the remaining ring atoms are carbon. It is preferably 7 to 10 yuan. E.g:

It can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups according to the number of constituent rings, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic.

"Optional" or "optionally" means that the subsequently described events or conditions may, but need not necessarily occur, and that the description includes situations in which the events or conditions occur and situations in which the events or conditions do not occur.

The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include heavy hydrogen and hydrogen variants, as long as the valence state of the particular atom is normal and the compound after substitution is stable of. When the substituent is a keto group (ie = O), it means that two hydrogen atoms are substituted. Ketone substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted. Unless otherwise specified, the types of substituents and trees may be arbitrary on the basis that they are chemically achievable.

When any variable (such as R) appears more than once in the composition or structure of a compound, its definition in each case is independent.

The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms that are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues, and No excessive toxicity, irritation, allergic reactions or other problems or function concurrently, commensurate with a reasonable benefit / risk ratio.

"Pharmaceutically acceptable salts" means those salts that retain the biological effectiveness and properties of the parent compound. Such salts include: (1) Forming a salt with an acid, which is obtained by reacting the free base of the parent compound with an inorganic acid or an organic acid. The inorganic acid includes hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, and sulfurous acid And perchloric acid, organic acids include acetic acid, trifluoroacetic acid, propionic acid, acrylic acid, hexanoic acid, cyclopentane propionic acid, glycolic acid, pyruvic acid, oxalic acid, (D) or (L) malic acid, fumaric acid , Maleic acid, benzoic acid, hydroxybenzoic acid, γ-hydroxybutyric acid, methoxybenzoic acid, phthalic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid , P-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, cinnamic acid, dodecyl sulfuric acid, gluconic acid, glutamic acid, aspartic acid, stearic acid, mandelic acid, succinic acid or propionic acid Diacids, etc. (2) The salts of acid protons present in the parent compound are replaced by metal ions or coordinated with organic bases. Examples of metals are alkali metal ions, alkaline earth metal ions, or aluminum ions. Organic bases are ethanolamine, diethanolamine, and triethanolamine. Ethanolamine, tromethamine, N-methylglucamine, etc.

In addition to salt forms, the compounds provided by the invention also exist in prodrug forms. The prodrugs of the compounds described herein easily undergo chemical changes under victory conditions to transform the compounds of the invention. In addition, prodrugs can be converted to the compounds of the invention by chemical or biochemical methods in the in vivo environment.

Certain compounds of the invention may exist in unsolvated or solvated forms, including hydrated forms. Generally speaking, the solvated form is equivalent to the unsolvated form and is included in the scope of the present invention.

Unless otherwise stated, use deed keys and imaginary keys

Represents the absolute configuration of a three-dimensional center, use

Represents the relative configuration of a three-dimensional center. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, unless specified otherwise, they include E, Z geometric isomers. Likewise, all tautomeric forms are included within the scope of the present invention.

The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)-and (+)-enantiomers, (R)-and (S) -enantiomers, non- Enantiomers, (D) -isomers, (L) -isomers, and their racemic and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of these Mixtures are within the scope of the present invention. Substituents such as alkyl groups may have additional asymmetric carbon atoms. All these isomers and mixtures thereof are included in the scope of the present invention.

The optically active (R)-and (S) -isomers and (D)-and (L) -isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If an enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, where the resulting mixture of diastereomers is separated and the auxiliary group is cleaved to provide pure The desired enantiomer. Alternatively, when the molecule contains a basic group (such as an amino group) or an acidic functional group (such as a carboxylic acid), a salt of a diastereomer is formed with an appropriate optically active acid or base, and then known by the art The conventional method is used for resolution, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which employs a chiral stationary phase and is optionally combined with chemical derivatization methods (for example, the formation of carbamate from amines) Acid salt).

The compound of the present invention may contain unnatural proportions of atomic isotopes in one or more atoms constituting the compound. For example, compounds can be labeled with radioactive isotopes, such as tritium, iodine-125, C-14. All changes in isotopic composition of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.

"Pharmaceutical composition" refers to combining one or more of the compounds of the present invention or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof with other chemical components, such as a pharmaceutically acceptable carrier, carrier Form or diluent mixed. The purpose of the pharmaceutical composition is to facilitate the process of administration to animals.

"Pharmaceutically acceptable carrier" means an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the administered compound, such as but not limited to: calcium carbonate, calcium phosphate , Various sugars (such as lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic polymer or methacrylic polymer, gel, water, polyethylene glycol, Propylene glycol, ethylene glycol, castor oil or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil, etc.

Excipient refers to an inert substance added to the pharmaceutical composition to further facilitate administration of the compound. Examples of excipients include (but are not limited to) calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oil, and polyethylene glycol.

In addition to the pharmaceutically acceptable carrier, the aforementioned pharmaceutical composition may also include adjuvants commonly used in medicine (agents), for example: antibacterial agents, antifungal agents, antimicrobial agents, shelf-stable agents, Colorants, solubilizers, thickeners, surfactants, complexing agents, proteins, amino acids, fats, sugars, vitamins, minerals, trace elements, sweeteners, colors, flavors, or combinations thereof.

The compounds of the present invention can be prepared by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the implementation methods formed by the combination with other chemical synthetic methods, and the well known to those skilled in the art Equivalently, preferred embodiments include but are not limited to the embodiments of the present invention.

The solvent used in the present invention is commercially available. The present invention uses the following abbreviations:

Compounds are named manually or by ChemDraw software, and commercially available compounds use the supplier catalog name.

LCMS gradient general formula: AxBy or CxDy (x + y = 100), A, C: water (acid: containing 0.1% formic acid, basic: containing 0.1% ammonium bicarbonate), B, D: acetonitrile (acid: containing 0.1 % Formic acid, alkaline: containing 0.1% ammonium bicarbonate), gradient conditions: acetonitrile y% ~ 95%. The total elution time is shown in each spectrum (2-10 minutes).

HPLC gradient general formula: AxBy or CxDy (x + y = 100), A, C: water (acid: containing 0.1% trifluoroacetic acid, basic: containing 0.1% ammonium bicarbonate), B, D: acetonitrile (acid: Containing 0.1% trifluoroacetic acid, alkaline: containing 0.1% ammonium bicarbonate), gradient conditions: acetonitrile y% ~ 95%. The total elution time is shown in each spectrum (generally 16 minutes).

detailed description

The invention will be described in detail by the following examples, but it does not mean any adverse limitation of the invention. The present invention has been described in detail herein, and its specific embodiment modes are also disclosed. For those skilled in the art, various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention Will be obvious.

Intermediate Preparation Example 1: Preparation of Intermediate D1

At room temperature, the compound 4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (5.0 g, 24.8 mmol) and triethylamine (7.54 g, 74.5 mmol) were dissolved in methylene chloride (50 mL), and then added Methanesulfonyl chloride (3.13g, 27.3mmol) was slowly added dropwise. After the dropwise addition, the reaction mixture was stirred at room temperature for 8 hours. TLC (PE: EtOAc = 1: 1, Rf = 0.6) showed that the starting material disappeared and the product was formed. The reaction solution was diluted with water (40 mL), and the mixture was extracted with ethyl acetate (350 mL × 2). The organic phases were combined and dried with anhydrous Na ₂ SO ₄ and filtered, and the filtrate was concentrated under reduced pressure to obtain the target crude product D1 (6.37 g, yield: 91.7%, white solid).

¹ H NMR (400 MHz, CD ₃ Cl ₃ ) δ: 4.89-4.87 (m, 1H) 3.73-3.67 (m, 2H), 3.33-3.26 (m, 2H), 3.09 (s, 3H), 1.97-1.94 ( m, 2H), 1.83-1.79 (m, 2H), 1.41 (s, 9H).

Intermediate Preparation Example 2: Preparation of Intermediate D2

In a 250 mL single-necked bottle, compound D1 (3.00 g, 10.7 mmol), compound 4-nitropyrazole (1.28 g, 11.3 mmol), K ₂ CO ₃ (2.97 g, 21.5 mmol), and DMF (40 mL) were added sequentially. After the reaction system was stirred uniformly, the temperature was raised to 110 ° C and the reaction was stirred for 1 hour. TLC (PE: EtOA c = 3: 1, Rf = 0.5) showed that the starting material disappeared and the product was formed. The reaction solution was diluted with water (100 mL), and the mixture was extracted with ethyl acetate (50 mL × 2). The organic phases were combined and washed with saturated sodium chloride solution (100 mL X 3). The resulting organic solution was dried and filtered with anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure to obtain the target crude product D2 (1.94 g, yield: 61%, white solid).

¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.17 (s, 1H), 8.08 (s, 1H), 4.30-4.26 (m, 3H) 2.82-2.93 (m, 2H), 2.18-2.15 (m, 2H) , 1.93-1.89 (m, 2H), 1.46 (s, 9H).

Intermediate Preparation Example 3: Preparation of Intermediate D3

After dissolving compound D2 (1.00 g, 3.37 mmol) in THF (10 mL) in a 100 mL three-necked flask, the solution was cooled to -78 ° C, and then LiHMDS (8.76 mL, 8.76 mmol) was added dropwise to the reaction solution. The reaction solution was stirred at -78 ° C for 30 min, and then hexachloroethane (1.20 g, 5.06 mmol) in THF (10 mL) was slowly added to the reaction solution. After the dropwise addition, the mixture was stirred at -78 ° C for 2.5 hours. TLC (PE: EtOAc = 3: 1, Rf = 0.7) showed that the starting material disappeared and the product was formed. The reaction was quenched with saturated aqueous ammonium chloride solution (10 mL), the mixture was extracted with ethyl acetate (50 mL × 3), the organic phases were combined, dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient: PE: EtOAc = 100: 1 to 5: 1) to obtain the target product D3 (980 mg, yield: 87.5%, white solid).

¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.18 (s, 1H), 4.50-4.3 (m, 3H), 2.82-2.93 (m, 2H), 2.18-2.15 (m, 2H), 1.93-1.89 (m , 2H), 1.46 (s, 9H).

Intermediate Preparation Example 4: Preparation of Intermediate D4

Compound D3 (500 mg, 1.69 mmol), methyl borate (1.01 g, 16.9 mmol), K ₂ CO ₃ (699 mg, 5.06 mmol), Pd (dppf) Cl ₂ (123 mg, 0.170 mmol) were added to 1,4- In a mixed solvent of Dioxane (5 mL) and H ₂ O (1 mL), nitrogen was replaced three times. The temperature of the reaction solution was raised to 85 ° C, and the reaction was stirred for 10 hours. LCMS detected compound D4 and D3 disappeared completely. The reaction solution was cooled to room temperature and added to a mixed solution of water (10 mL) and ethyl acetate (30 mL). After the two phases were separated, the aqueous phase was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, dried and concentrated, and the residue was purified by column chromatography (eluent gradient PE: EtOAc = 100: 1-5: 1) to obtain the target product D4 (290 mg, yield: 55.4%, white solid).

¹ H NMR (400 MHz, CD ₃ Cl ₃ ) δ 8.09 (s, 1H), 4.32-4.17 (m, 3H), 2.87-2.85 (m, 2H), 2.69 (s, 2H), 2.19-2.10 (m , 2H), 1.88 (d, J = 12Hz, 2H), 1.48 (s, 9H).

Intermediate Preparation Example 5: Preparation of Intermediate D5

Pd / C (100 mg, 10%) was added to a solution of compound D4 (240 mg, 0.770 mmol) in ethanol (5 mL) at room temperature. The reaction mixture was filled with hydrogen and replaced with vacuum three times, and then the system was stirred at 60 ° C for 1 hour under hydrogen atmosphere, and the reaction was completed by LCMS. The reaction mixture was filtered to remove Pd / C. The filtrate was concentrated under reduced pressure to obtain crude target compound D5 (200 mg, yield: 93%, brownish yellow solid).

LCMS: Rt: 1.178 min; MS m / z (ESI): 281.3 [M + H].

Intermediate Preparation Example 6: Preparation of Intermediate D6

Potassium carbonate (8.8g, 64.4mmol) was added to compound 4-nitro-1pyrazole (3.6g, 32.2mmol) and compound (4-hexahydropyran) methanesulfonate (5.8g, 32.2mmol) at room temperature ) In DMF (50 mL) solution, the reaction solution was then heated to 80 ° C. and stirred overnight. After the reaction was completed, the reaction solution was diluted with water (300 mL), the mixture was extracted with ethyl acetate (100 mL X 3), the organic phase was washed with saturated aqueous sodium chloride solution (300 mL X 3); then the organic solution was dried and filtered with anhydrous sodium sulfate, The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (elution gradient: PE: EtOAc = 5: 1) to obtain the target product D6 (2.7 g, yield: 43%, yellow solid).

Intermediate Preparation Example 7: Preparation of Intermediate D7

After cooling the compound D6 (0.2 g, 1.01 mmol) in THF (5 mL) to -78 ° C, LiHMDS (2 mL, 1 M) was slowly added dropwise; after the completion of the dropwise addition, the reaction solution was continuously stirred for 20 minutes, and then added to the reaction solution. Ethyl chloride (0.288 g, 1.22 mmol); the mixture was stirred at -78 ° C for 1 hour. TLC (PE: EtOAc = 3: 1, Rf = 0.7) showed that the starting material disappeared and the product was formed. The reaction was quenched with saturated aqueous ammonium chloride solution (10 mL), the mixture was extracted with ethyl acetate (50 mL × 3), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain the target crude product D7 (130 mg, yield: 55%, yellow solid).

¹ H NMR (400 MHz, DMSO-d6) δ: 8.50 (s, 1H), 4.70-4.60 (m, 1H) 3.97 (d, J = 8.4 Hz, 2H), 3.53-3.47 (m, 2H), 2.01- 1.85 (m, 4H).

Intermediate Preparation Example 8: Preparation of Intermediate D8

Under nitrogen protection, compound D7 (300mg, 1.5mmol) and methylboronic acid (448.9mg, 7.5mmol) were dissolved in dioxane (10mL) and water (2mL), and then Pd (dppf) Cl ₂ (110 mg, 0.15 mmol) and potassium phosphate (955 mg, 4.5 mmol); the reaction system was protected by nitrogen and heated to 90 ° C and stirred under this condition for 16 hours. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed with ethyl acetate (20 mL). The organic phases were combined, dried and filtered, and the filtrate was concentrated under reduced pressure; the residue was subjected to column chromatography (elution gradient: PE: EtOAc = 10) : 1-5: 1) Purification to obtain the target product D8 (220mg, yield: 80.3%, light yellow solid).

¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.04 (s, 1H), 4.25-4.19 (m, 1H), 4.10-4.06 (m, 2H), 3.55-3.44 (m, 2H), 2.62 (s, 3H ), 2.32-2.22 (m, 2H), 1.78-1.52 (m, 2H)

Intermediate Preparation Example 9: Preparation of Intermediate D9

In a 100 mL single-necked bottle, compound D8 (220 mg, 1.04 mmol), MeOH (50 mL), and Pd / C (22 mg, 5%) were added sequentially. The reaction system was stirred at room temperature for 16 hours under a hydrogen atmosphere. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd / C, and the filtrate was concentrated under reduced pressure to obtain the target crude product D9 (180 mg, yield: 96%, brown solid). The crude product was used directly in the next reaction.

LCMS: Rt: 0.365 min; MS m / z (ESI): 182.4 [M + H].

Intermediate Preparation Example 10: Preparation of Intermediate D10

Add the compound (2-aminophenyl) dimethyl phosphine oxide (0.5g, 3.27mmol), 2,6-dichloro-5- (trifluoromethyl) pyrimidine (847mg, 3.92) to a 100mL three-necked bottle mmol), DIEA (1.26 g, 9.81 mmol) and DMF (10 mL). The mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction mixture was poured into water (50 mL). The mixture was extracted with EtOAc (30 mL 2), the organic phases were combined and washed with saturated aqueous sodium chloride solution (60 mL X 2). The resulting organic solution was dried and filtered with anhydrous Na ₂ SO ₄ , and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient: PE: EtOAc = 5: 1-0: 1) to obtain a mixture of the target product D10 and the by-product D10 ′ (0.9 g, yield: 87.5%, light yellow solid). The mixture was difficult to separate and was used directly in the next reaction.

LCMS: Rt: 1.245min, 1.521min; MS m / z (ESI): 350.1 [M + H].

Intermediate Preparation Example 11: Preparation of Intermediate D11

Compound D6 (200 mg, 1.0 mmol) was dissolved in MeOH (20 mL) at room temperature, and then Pd / C (40 mg) was added; the reaction system was replaced with hydrogen three times and the reaction was stirred at room temperature for 2 h. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd / C, and the filtrate was concentrated under reduced pressure to obtain the target crude product D11 (100 mg, yield: 58.8%, yellow oily liquid). The crude product was used directly in the next reaction.

LCMS: Rt: 0.273 min; MS m / z (ESI): 168.4 [M + H].

Intermediate Preparation Example 12: Preparation of Intermediate D12

The compound (2-aminophenyl) dimethyl phosphine oxide (1.0 g, 5.0 mmol), 2,4,5-trichloropyrimidine (923 mg, 5.0 mmol), NaHCO ₃ (1.59 g, 15.0 mmol) were dissolved at room temperature In EtOH / H ₂ O (45 mL, 8: 1), the reaction system was heated to 80 ° C. under the protection of Ar and the reaction was stirred overnight. LCMS detected that the raw material was half reacted, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined and washed with saturated NaCl solution (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated. The residue was purified through a silica gel column (elution gradient: PE: EtOAC = 1: 10-1: 5) to obtain the target intermediate D12 (740 mg, yield: 39.7%, white solid).

LCMS: Rt: 1.827 min; MS m / z (ESI): 316.0 [M + H].

Intermediate Preparation Example 13: Preparation of Intermediate D13

At room temperature, the compounds 4-nitropyrazole (3.39g, 30mmol), 2-bromoethanol (3.75g, 30mmol), K ₂ CO ₃ (4.97g, 36mmol) and CH ₃ CN ( 50mL). The mixture was heated to 80 ° C and reacted for 16 hours. After the reaction, the reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target product D13 (4.7 g, yield: 100%, white solid), which was directly used in the next step.

LCMS: Rt: 0.775 min; MS m / z (ESI): 158.1 [M + H].

Intermediate Preparation Example 14: Preparation of Intermediate D14

At room temperature, D13 (4.7g, 30mmol), TBSCl (5.0g, 33mmol), imidazole (2.5g, 36mmol) and dichloromethane (50mL) were added to a 250mL single-necked bottle in this order. The reaction solution was stirred at room temperature for 16 hours. After the reaction, the reaction solution was washed with water (40 mL × 3), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was subjected to column chromatography (eluent gradient: PE: EtOAC = 20: 1- 10: 1) Purification to obtain the target product D14 (6.7 g, yield: 80%, white solid).

LCMS: Rt: 1.399 min; MS m / z (ESI): 272.2 [M + H].

Intermediate Preparation Example 15: Preparation of Intermediate D15

At -60 ° C, LiHMDS (25 mL, 25 mmol) was slowly added to a solution of compound D14 (2.7 g, 10 mmol) in THF (30 mL). After the addition was complete, the reaction solution was stirred at -60 ° C for 30 minutes, and then C ₂ Cl ₆ (3.0 g, 12 mmol) was added in portions. After the addition, the reaction solution was stirred at -60 ° C for 3 hours. After the reaction was completed, the reaction solution was heated to 0 ° C, and then quenched with saturated aqueous ammonium chloride solution (20 mL). After the two phases were separated, the aqueous phase was extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with water (50 mL) and saturated brine (50 mL), then dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient: PE: EtOAC = 20: 1-10: 1) to obtain the target product D15 (1.0 g, yield: 30%, yellow solid).

LCMS: Rt: 1.553 min; MS m / z (ESI): 306.2 [M + H].

¹ H NMR (400 MHz, DMSO) δ: 8.61 (s, 1H), 4.43 (t, J = 4.0 Hz, 2H), 4.05 (t, J = 4.0 Hz, 2H), 0.84 (s, 9H), 0.00 ( s, 6H).

Intermediate Preparation Example 16: Preparation of Intermediate D16

At room temperature, the compound 4-nitropyrazole (2.26g, 20mmol), 1-bromo-2-methoxyethane (2.76g, 20mmol), K ₂ CO ₃ (6.72g, 40 mmol) and CH ₃ CN (30 mL). The mixture was heated to 80 ° C and stirred for 16 hours. The reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target product D16 (3.1 g, yield: 91.1%, colorless liquid), which was directly used in the next step.

LCMS: Rt: 0.984 min; MS m / z (ESI): 172.4 [M + H].

Intermediate Preparation Example 17: Preparation of Intermediate D17

At room temperature, compound D16 (342.0 mg, 2 mmol), MeOH (5 mL) and Pd / C (34.2 mg, 10%) were added in sequence in a 100 mL single-necked bottle. The reaction system was stirred at room temperature for 16 hours under a hydrogen atmosphere. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd / C, and the filtrate was concentrated under reduced pressure to obtain the crude target product D17 (284 mg, yield: 100%, yellow liquid). The crude product was used directly in the next reaction.

LCMS: Rt: 0.327 min; MS m / z (ESI): 142.2 [M + H].

Intermediate Preparation Example 18: Preparation of Intermediate D18

At room temperature, compound D14 (271 mg, 1.0 mmol), MeOH (5 mL) and Pd / C (27.1 mg, 10%) were added in sequence in a 100 mL single-necked bottle. The reaction system was stirred at room temperature for 16 hours under a hydrogen atmosphere. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd / C, and the filtrate was concentrated under reduced pressure to obtain the crude target product D18 (241 mg, yield: 100%, black liquid). The crude product was used directly in the next reaction.

LCMS: Rt: 0.827 min; MS m / z (ESI): 242.4 [M + H].

Intermediate Preparation Example 19: Preparation of Intermediate D19

LiHMDS (15 mL, 15 mmol) was slowly added to a solution of compound D16 (1.71 g, 10 mmol) in THF (30 mL) at -60 ° C. After the addition was complete, the reaction solution was stirred at -60 ° C for 30 minutes, and then C ₂ Cl ₆ (4.68 g, 20 mmol) was added in portions. After the addition, the reaction solution was stirred at -60 ° C for 4 hours. After the reaction was completed, the reaction solution was heated to 0 ° C, and then quenched with saturated aqueous ammonium chloride solution (20 mL). After the two phases were separated, the aqueous phase was extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with saturated brine (30 mL), then dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient: PE: EtOAC = 10: 1-5: 1) to obtain the target product D19 (1.8 g, yield: 87.4%, yellow liquid).

LCMS: Rt: 1.068 min; MS m / z (ESI): 206.0 [M + H].

Intermediate Preparation Example 20: Preparation of Intermediate D20

Compound D19 (205 mg, 1.0 mmol), iron powder (280 mg, 5.0 mmol), NH ₄ Cl (428 mg, 8.0 mmol) were added to EtOH / H ₂ O (4: 1, 20 mL) at room temperature, and then the reaction solution Heat to 90 ° C for 2 hours. LCMS detected that the reaction was complete. The reaction solution was cooled to room temperature, and then saturated aqueous sodium bicarbonate solution (50 mL) and ethyl acetate (20 mL) were added to the reaction solution. The mixture was stirred for 5 minutes and then filtered. Extracted with ethyl acetate (20mL × 3); the organic phases were combined and washed with saturated aqueous sodium chloride solution (50mL), then dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain the target intermediate D20 (100mg, yield) : 57.1%, yellow liquid) directly used in the next step.

LCMS: Rt: 0.51 min; MS m / z (ESI): 176.2 [M + H].

Intermediate Preparation Example 21: Preparation of Intermediate D21

At room temperature, compound D12 (200 mg, 0.63 mmol), compound D5 (177 mg, 0.63 mmol) and NH ₄ Cl (101 mg, 1.9 mmol) were dissolved in EtOH (5 mL), and the reaction solution was stirred at 100 ° C. for 3 hours. After the reaction was detected by LCMS, the reaction solution was cooled to room temperature, the mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient PE / EtOAc = 1: 100, Rf = 0.3) to obtain the target Compound D21 (180 mg, yield: 50.8%, white solid).

LCMS: Rt: 1.42 min; MS m / z (ESI): 560.3 [M + H].

Intermediate Preparation Example 22: Preparation of Intermediate D22

After cooling the solution of compound D6 (0.2g, 1.01mmol) in THF (5mL) to -78 ° C, LiHMDS (2mL, 1M) was slowly added dropwise; after the completion of the dropwise addition, the reaction solution was continuously stirred for 20 minutes. Ethyl chloride (0.288 g, 1.22 mmol); the mixture was stirred at -78 ° C for 1 hour. TLC (PE: EtOAc = 3: 1, Rf = 0.7) showed that the starting material disappeared and the product was formed. The reaction was quenched with saturated aqueous ammonium chloride solution (10 mL), the mixture was extracted with ethyl acetate (50 mL × 3), the organic phase was dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain the target crude product D22 (130 mg, yield: 55%, yellow solid).

¹ H NMR (400 MHz, DMSO-d6) δ: 8.50 (s, 1H), 4.70-4.60 (m, 1H) 3.97 (d, J = 8.4 Hz, 2H), 3.53-3.47 (m, 2H), 2.01- 1.85 (m, 4H).

Intermediate Preparation Example 23: Preparation of Intermediate D23

Compound D22 (150 mg, 0.64 mmol), iron powder (180 mg, 3.2 mmol) and NH ₄ Cl (274 mg, 5.12 mmol) were added to EtOH / H ₂ O (4: 1, 20 mL) at room temperature, and the reaction system was heated up Reaction to 90 ° C for 2h. LCMS detected the reaction was complete. The reaction solution was cooled to room temperature and diluted with the addition of saturated aqueous sodium bicarbonate solution (50 mL) and ethyl acetate (20 mL); the mixture was stirred for 5 minutes and filtered. After the two phases were separated, the aqueous phase was separated with ethyl acetate (20 mL X 3) Extraction, the organic phases are combined, washed with saturated aqueous sodium chloride solution (50 mL), then dried and filtered with anhydrous sodium sulfate, and the filtrate is concentrated under reduced pressure to obtain the target intermediate D23 (100 mg, yield: 76.9%, yellow liquid) ) Used directly in the next step.

LCMS: Rt: 0.47 min; MS m / z (ESI): 202.1 [M + H].

Intermediate Preparation Example 24: Preparation of Intermediate D24

Under nitrogen protection, dissolve compound D15 (305mg, 1.0mmol) and methylboronic acid (360.0mg, 6.0mmol) in dioxane (10mL) and water (2mL), then add Pd (dppf) Cl to the system ₂ (73.3 mg, 0.1 mmol) and potassium carbonate (276 mg, 2.0 mmol). After the reaction system was protected with nitrogen, the temperature was raised to 100 ° C and the reaction was stirred under this condition for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with ethyl acetate (20 mL), the organic phases were combined, dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure; the residue was subjected to column chromatography (elution gradient: PE: EtOAc = 10: 1-5: 1) purification to obtain the target product D24 (160 mg, yield: 56.1%, light yellow liquid).

LCMS: Rt: 1.863 min; MS m / z (ESI): 286.4 [M + H].

Intermediate Preparation Example 25: Preparation of Intermediate D25

In a 100 mL single-necked bottle, compound D24 (160 mg, 0.56 mmol), MeOH (5 mL), and Pd / C (16 mg, 10%) were added in sequence. The reaction system was stirred at room temperature for 16 hours under a hydrogen atmosphere. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd / C, and the filtrate was concentrated under reduced pressure to obtain the crude target product D25 (143 mg, yield: 100%, yellow liquid). The crude product was used directly in the next reaction.

LCMS: Rt: 0.890 min; MS m / z (ESI): 256.3 [M + H].

Intermediate Preparation Example 26: Preparation of Intermediate D26

Compound D19 (500 mg, 2.5 mmol) and methylboronic acid (900 mg, 15.0 mmol) were dissolved in 1,4-dioxane (36 mL) at room temperature, and then K ₂ CO ₃ (1.04 g, 7.5 mmol) was added dropwise Water (4mL) solution; after replacing the reaction system with argon three times, Pd (dppf) Cl ₂ (200mg, 0.25mmol) was added. After replacing the system with argon three times, the system was heated to 80 ° C and stirred overnight. The reaction was completed by LCMS. The reaction solution was cooled to room temperature. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (30 mL X 3). The organic phases were combined and washed with saturated aqueous sodium chloride solution (100 mL). The solution was dried with anhydrous sulfuric acid. Sodium was dried and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by thin layer chromatography (eluent gradient: PE: ETOAC = 2: 1) to obtain the target intermediate D26 (170 mg, Yeild: 36.8%, yellow oily liquid ).

LCMS: Rt: 1.383 min; MS m / z (ESI): 186.4 [M + H].

Intermediate Preparation Example 27: Preparation of Intermediate D27

Compound D26 (170 mg, 0.92 mmol) was dissolved in MeOH (20 mL) at room temperature, and then Pd / C (30 mg) was added; the reaction system was replaced with hydrogen three times, and the reaction was stirred at room temperature for 2 hours. The reaction was completed by LCMS. The reaction solution was filtered to remove Pd / C. The filtrate was concentrated under reduced pressure to obtain the target crude product D27 (155 mg, yield: 100%, yellow oily liquid). The crude product was used directly in the next reaction.

LCMS: Rt 0.282min; MS M / z (ESI): 156.3 [M + H].

Intermediate Preparation Example 28: Preparation of Intermediate D28

Compound E7 (390 mg, 0.79 mmol) and DIETOAC (308 mg, 2.36 mmol) were dissolved in DMF (15 mL) at room temperature, then methyl bromoacetate (121 mg, 0.79 mmol) was added to the reaction solution, and the reaction solution was stirred at room temperature React for 2 hours. After LCMS detected that the reaction was complete, the reaction mixture was poured into saturated aqueous ammonium chloride solution (20 mL). The mixture was extracted with ethyl acetate (30 mL × 4), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient: EtOAc: MeOH = 10: 1, Rf = 0.5) to obtain the target product D28 (366 mg, yield: 87%, beige solid).

LCMS: Rt: 0.77 min; MS m / z (ESI): 532.4 [M + H].

Intermediate Preparation Example 29: Preparation of Intermediate D29

Under nitrogen protection, compound D23 (300 mg, 1.5 mmol) and methylboronic acid (448.9 mg, 7.5 mmol) were dissolved in dioxane (10 mL) and water (2 mL), and then Pd (dppf) Cl was added to the system ₂ (110 mg, 0.15 mmol) and potassium phosphate (955 mg, 4.5 mmol); the reaction system was protected by nitrogen and heated to 90 ° C and stirred under this condition for 16 hours. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed with ethyl acetate (20 mL). The organic phases were combined, dried and filtered, and the filtrate was concentrated under reduced pressure; the residue was subjected to column chromatography (elution gradient: PE: EtOAc = 10) : 1-5: 1) to obtain the target product D29 (220 mg, yield: 80.3%, light yellow solid).

¹ H NMR (400 MHz, CDCl ₃ ) δ: 8.04 (s, 1H), 4.25-4.19 (m, 1H), 4.10-4.06 (m, 2H), 3.55-3.44 (m, 2H), 2.62 (s, 3H ), 2.32-2.22 (m, 2H), 1.78-1.52 (m, 2H)

Intermediate Preparation Example 30: Preparation of Intermediate D30

Compound D22 (120 mg, 0.51 mmol), cyclopropylboronic acid (263 mg, 3.06 mmol) was dissolved in 1,4-dioxane (18 mL), and 2 mL of K ₂ CO ₃ (211 mg, 1.53 mmol) solution was added dropwise, Ar Vent 3 times, add Pd (dppf) Cl ₂ (38mg, 0.051mmol), fully ventilate 3 times, heat to 80 ° C for overnight. LCMS detected that the basic reaction was complete. The reaction solution was cooled to room temperature, poured into a separatory funnel and added with water (50 mL) and ETOAC (20 mL × 3) for extraction. The combined organic phases were washed with saturated NaCl solution (50 mL) and passed through anhydrous Na ₂ SO ₄ After drying and filtering, the filtrate was concentrated and evaporated to dryness, and the crude product was purified by TLC preparation plate (PE: ETOAC = 2: 1) to obtain the target intermediate D30 (120 mg, yield: 100%, yellow oily liquid).

LCMS: Rt: 1.553 min; MS m / z (ESI): 238.4 [M + H].

Intermediate Preparation Example 31: Preparation of Intermediate D31

Compound D30 (120 mg, 0.51 mmol) was dissolved in MeOH (20 mL) at room temperature, and then Pd / C (20 mg) was added; after the addition, the reaction system was replaced with hydrogen three times, and then the system was stirred at room temperature for 2 h. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd / C, and the filtrate was concentrated under reduced pressure to obtain the target crude product D31 (80 mg, yield: 100%, yellow oily liquid). The crude product was used directly in the next reaction.

LCMS: Rt: 0.542 min; MS m / z (ESI): 208.3 [M + H].

Intermediate Preparation Example 32: Preparation of Intermediate D32

The compound (507 mg, 3.0 mmol), 5-fluoro-2,4-dichloropyrimidine (501 mg, 3.0 mmol) and NaHCO ₃ (756 mg, 9.0 mmol) were dissolved in EtOH / H ₂ O (8: 1, 45 mL) at room temperature ), The reaction system was then heated to 80 ° C and reacted overnight. After the reaction, the reaction solution was cooled to room temperature and diluted with water (100 mL). The mixture was extracted with ethyl acetate (50 mL X 3). The organic phases were combined and washed with saturated aqueous sodium chloride solution (100 mL), and then dried over anhydrous sodium sulfate. Filtration, the filtrate was concentrated under reduced pressure, the residue was prep-HPLC (eluent gradient:

) Purification to obtain the target intermediate D32 (36 mg, yield: 4%, white solid).

LCMS: Rt: 1.453 min; MS m / z (ESI): 300.3 [M + H].

Intermediate Preparation Example 33: Preparation of Intermediate D33

A mixture of 5-bromo-2,4-dichloropyrimidine compound (5g, 22.02mmol), compound 2- (aminophenyl) dimethyl phosphine oxide (3.7g, 22.02mmol) and NaHCO ₃ (5.5g, 66.06 mmol) was dissolved in EtOH (50 mL) and water (10 mL). The reaction solution was heated to 100 ° C and stirred for 10 hours, and the reaction was completed by LCMS. The reaction solution was cooled to room temperature and diluted with water (50 mL). The mixture was extracted with dichloromethane (50 mL × 5). The organic phase was dried and filtered with sodium sulfate. The filtrate was concentrated under reduced pressure. Gradient PE / EtOAc = 100: 1 to 1: 100) purification to obtain the target compound D33 (4g, yield: 50%, white solid).

LCMS: Rt: 1.23 min; MS m / z (ESI): 362.1 [M + H].

Intermediate Preparation Example 34: Preparation of Intermediate D34

Compound (2-aminophenyl) dimethyl phosphine oxide (450 mg, 2.5 mmol), compound 2,4-dichloro-5-methoxypyrimidine (505 mg, 3.0 mmol) and DIETOAC (650 mg, 5.0 mmol) at room temperature ) Was dissolved in DMF (10 mL), and the reaction solution was stirred at 140 ° C. for 12 hours. After LCMS detected that the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the residue was purified by preparative TLC (eluent gradient ETOAC / MeOH = 10: 1) to obtain the target product D34 (340 mg, yield: 45%, light yellow solid ).

LCMS: Rt: 1.28 min; MS m / z (ESI): 312.2 [M + H].

Intermediate Preparation Example 35: Preparation of Intermediate D35

Compound 2,4-dichloro-5-methylpyrimidine (500 mg, 3.07 mmol) and compound (2-aminophenyl) dimethyl phosphine oxide (466 mg, 2.76 mmol) were dissolved in DMF (5 mL) at room temperature, then DIEA (1.18 mg, 9.21 mmol) was added. The reaction solution was heated to 100 ° C and stirred for 10 hours. The reaction was completed by LCMS. The reaction solution was cooled to room temperature and diluted with water (20 mL). The mixture was extracted with dichloromethane (20 mL × 3). The organic phase was dried and filtered over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure. The residue was passed through a thin layer. Chromatography (eluent gradient PE / EtOAc = 1: 1, R _f = 0.2) purification gave the target compound D35 (200 mg, yield: 22%, white solid).

LCMS: Rt: 1.20 min; MS m / z (ESI): 296.1 [M + H].

Intermediate Preparation Example 36: Preparation of Intermediate D36

Compound E7 (390 mg, 0.79 mmol) and DIEA (308 mg, 2.36 mmol) were dissolved in DMF (15 mL) at room temperature, then methyl bromoacetate (121 mg, 0.79 mmol) was added to the reaction solution, and the reaction solution was stirred at room temperature React for 2 hours. After LCMS detected that the reaction was complete, the reaction mixture was poured into saturated aqueous ammonium chloride solution (20 mL). The mixture was extracted with ethyl acetate (30 mL × 4), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient: EtOAc: MeOH = 10: 1, Rf = 0.5) to obtain the target product D36 (366 mg, yield: 87%, beige solid).

LCMS: Rt: 0.77 min; MS m / z (ESI): 532.4 [M + H].

Intermediate Preparation Example 37: Preparation of Intermediate D37

At room temperature, compound D36 (106 mg, 0.2 mmol) was dissolved in MeOH / THF / H2O (4 mL / 4 mL / 1 mL) and LiOH (25 mg, 0.6 mmol) was added, and the reaction solution was stirred at room temperature for 3 hours. After LCMS detected that the reaction had been completed, the pH of the reaction mixture was adjusted to pH ~ 3, and then the reaction solution was directly concentrated under reduced pressure to obtain the crude target product D37 (104 mg, yield: 100%, white solid).

LCMS: Rt: 0.29 min; MS m / z (ESI): 518.0 [M + H].

Intermediate Preparation Example 38 and Intermediate Preparation Example 39: Preparation of Intermediate D38 and D39

At room temperature, the compound 2,4-dichloro-5-trifluoromethylpyrimidine (2.2g, 10mmol), compound (2-aminophenyl) dimethyl phosphine oxide (2.0g, 12mmol) and DIEA (4.0g, 30 mmol) was dissolved in DMF (25 mL), and the reaction solution was stirred at room temperature for 16 hours. LCMS detected the disappearance of the raw materials, and at the same time, the production of two isomers. The reaction solution was concentrated under reduced pressure, and the resulting product (crude product: 8.0 g) was separated by the following column chromatography conditions:

The first batch: 4.0g crude product, 330g reverse phase preparative column, eluent gradient: acetonitrile / water = 0-100%, time = 110 minutes; Peak 1: 28 minutes-38 minutes; Peak 2: 42 minutes-52 minutes .

The second batch: 4.0g crude product, 330g reverse phase preparative column, eluent gradient: acetonitrile / water = 0-100%, time = 130 minutes; Peak 1: 35 minutes to 41 minutes, Peak 2: 50 minutes to 55 minutes .

The two batches of Peak1 were combined and concentrated under reduced pressure to obtain product D38 (1.8 g, yield: 69%, yellow solid)

LCMS: Rt: 1.099min; MS m / z (ESI): 350.1 [M + H]

¹ H NMR (400 MHz, MeOD) δ: 11.1 (s, 1H), 8.67 (s, 1H), 8.21 (m, 1H), 7.68 (m, 2H), 7.32 (t, J = 8.0 Hz, 1H), 1.80 (s, 3H), 1.76 (s, 3H).

The two batches of Peak 2 were combined and concentrated under reduced pressure to obtain product D39 (0.7 g, yield: 69%, yellow solid)

LCMS: Rt: 1.358min; MS m / z (ESI): 350.1 [M + H],

¹ H NMR (400 MHz, MeOD) δ: 11.9 (s, 1H), 8.85 (s, 1H), 8.48 (m, 1H), 7.68 (m, 2H), 7.24 (t, J = 8.0 Hz, 1H), 1.83 (s, 3H), 1.79 (s, 3H).

Intermediate Preparation Example 40: Preparation of Intermediate D40

Compound D38 (350 mg, 1.0 mmol) and intermediate D5 (420 mg, 1.5 mmol) were dissolved in EtOH (30 mL) at room temperature. After adding NH ₄ Cl (535 mg, 10.0 mmol) to the reaction system, the reaction solution was heated to The reaction was stirred at 80 ° C for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with ethanol (10 mL X 2), the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient DCM: MeOH = 20: 1) to obtain Target intermediate D40 (420 mg, yield: 70.8%, white solid).

LCMS: Rt: 1.593 min; MS m / z (ESI): 594.5 [M + H].

Intermediate Preparation Example 41: Preparation of Intermediate D41

Compound D39 (350 mg, 1.0 mmol) and intermediate D5 (420 mg, 1.5 mmol) were dissolved in EtOH (30 mL) at room temperature. After adding NH ₄ Cl (535 mg, 10.0 mmol) to the reaction system, the reaction solution was heated to The reaction was stirred at 80 ° C for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with ethanol (10 mL X 2), the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient DCM: MeOH = 20: 1) to obtain Target intermediate D41 (450 mg, yield: 75.8%, white solid).

LCMS: Rt: 1.723 min; MS m / z (ESI): 594.6 [M + H].

Intermediate Preparation Example 42: Preparation of Intermediate D42

Compound D22 (200 mg, 0.86 mmol) and pinacol vinyl borate (800 mg, 5.18 mmol) were dissolved in 1,4-dioxane (18 mL) at room temperature, and then K ₂ CO ₃ (358 mg, after the addition of 2.59 mmol) in water (2mL) added and the reaction system was replaced 3 times with argon, _{Pd (dppf) Cl 2 (32mg} , 0.043mmol), the system then replaced 3 times, heating to 80 deg.] C overnight. The reaction was checked by LCMS. The reaction solution was cooled to room temperature and diluted with water (50 mL). The mixture was extracted with ethyl acetate (20 mL X 3). The combined organic phases were washed with saturated aqueous sodium chloride solution (50 mL) and then anhydrous sodium sulfate. It was dried and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by thin layer chromatography (eluent gradient: PE: ETOAC = 2: 1) to obtain the target intermediate D42 (170 mg, yield: 88.2%, yellow oily liquid) ).

LCMS: Rt: 1.502 min; MS m / z (ESI): 224.3 [M + H].

Intermediate Preparation Example 43: Preparation of Intermediate D43

To a solution of compound D42 (170 mg, 0.73 mmol) in MeOH (20 mL) was added Pd / C (30 mg) at room temperature, the reaction system was replaced with hydrogen three times, and the reaction was stirred at room temperature for 2 h. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd / C, and the filtrate was concentrated under reduced pressure to obtain the target crude product D43 (150 mg, yield: 100%, yellow oily liquid). The crude product was used directly in the next reaction.

LCMS: Rt: 0.303 min; MS m / z (ESI): 196.4 [M + H].

Intermediate Preparation Example 44: Preparation of Intermediate D44

At room temperature, compound 4-nitropyrazole (2.0g, 17.7mmol), compound 1,4-dioxaspiro [4.5] decan-8-yl methanesulfonate (4.17g, 17.7mmol) and potassium carbonate (4.9g, 35.4mmol) It was added to acetonitrile (50 mL), and the reaction system was heated to 80 ° C under the protection of argon and the reaction was stirred overnight. After the reaction was complete, the reaction mixture was concentrated under reduced pressure to remove acetonitrile, and the residue was diluted with water (50 mL) and ethyl acetate (50 mL); after the two phases were separated, the aqueous phase was extracted with ethyl acetate (20 mL x 2), and the organic phases were combined and used Dry filtration over anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient PE / ETOAC = 10/1) to obtain the target compound D44 (3g, yield: 67%, white) solid).

LCMS: Rt: 1.198 min; MS m / z (ESI): 254 [M + H].

Intermediate Preparation Example 45: Preparation of Intermediate D45

Under the protection of argon, the solution of compound D44 (2g, 8.66mmol) in THF (30mL) was cooled to -78 ° C, then LiHMDS (13mL, 1M, 13mmol) was slowly added dropwise to the system; the reaction solution was at this temperature after the addition The reaction was stirred for 30 minutes, and then a solution of hexachloroethane (4.1 g, 17.3 mmol) in THF (14 mL) was added dropwise. After the completion of the dropping, the reaction solution was stirred and reacted for 3 hours. After the reaction was completed, a saturated aqueous ammonium chloride solution (100 mL) was added to the reaction solution to quench the reaction, and the mixture was extracted with ethyl acetate (30 mL x 3); the organic phases were combined and dried with anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by thin layer chromatography (eluent gradient PE / EtOAc = 15/1 to 12/1) to obtain the target compound D45 (1.8 g, yield: 78%, white solid).

LCMS: Rt: 1.772 min; MS m / z (ESI): 288 [M + H].

Intermediate Preparation Example 46: Preparation of Intermediate D46

A mixture of compound D45 (1.5g, 5.23mmol), methyl boronic acid (5.5g, 91.7mmol), Pd ( dppf) Cl 2 (450mg, 0.61mmol) and sodium carbonate (5g, 47.2mmol) was added 1,4 -In a mixed solvent (20/4 mL) of dioxane and water, the reaction system was replaced with argon three times, then heated to 90 ° C. and stirred for 24 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and then the reaction solution was diluted with water (50 mL) and ethyl acetate (20 mL). After the two phases were separated, the aqueous phase was extracted with ethyl acetate (20 mL x 2); the organic phases were combined and used Dry filtration over anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient PE / ETOAC = 20/1 ～ 12/1) to obtain the target product D46 (1.2g, yield : 97%, white solid).

LCMS: Rt: 1.906min; MS m / z (ESI): 268 [M + H].

Intermediate Preparation Example 47: Preparation of Intermediate D47

At room temperature, compound D46 (1.2 g, 4.5 mmol) was dissolved in THF (20 mL) and Pd / C (content 10 wt%, 600 mg) was added. After the reaction system was replaced with hydrogen three times, it was stirred overnight at room temperature under a hydrogen atmosphere. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the target product D47 (1.1 g, yield: 98%, white solid).

LCMS: Rt: 0.373 min; MS m / z (ESI): 238 [M + H].

Intermediate Preparation Example 48: Preparation of Intermediate D48

At room temperature, compound D12 (900mg, 3.8mmol), compound D47 (640mg, 3.8mmol) and ammonium chloride (2.1g, 38mmol) were dissolved in ethanol (15mL), the reaction system was heated to 80 ℃ under argon protection 48 hours. After the reaction was completed, the reaction solution was cooled to room temperature, the mixture was filtered, the filtrate was concentrated under reduced pressure, the residue was dissolved with DCM / MeOH (20 / 1,100mL) and washed with water (30mL × 2), the organic phase was dried and filtered over anhydrous sodium sulfate The filtrate was concentrated under reduced pressure to obtain the target compound D48 (1.6 g, yield: 80%, red solid).

LCMS: Rt: 1.30 min; MS M / z (ESI): 517 [M + H].

Intermediate Preparation Example 49: Preparation of Intermediate D49

Under ice bath protection, dioxane hydrochloride (4M, 5 mL) solution was added to a solution of compound D48 (1.6 g) in methanol (5 mL), and the reaction solution was stirred at room temperature overnight. After the reaction was completed, the reaction solution was cooled to 0 ° C, and then saturated aqueous sodium bicarbonate solution was added to neutralize the reaction solution to pH> 7. The resulting mixture was concentrated under reduced pressure, and the residue was dissolved with DCM / MeOH (20 / 1,100 mL). 30mL x 3) Wash. The organic phase was dried and filtered with anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure, and the residue was subjected to column chromatography (eluent gradient DCM / MeOH = 40/1) to obtain the target compound D49 (1.4 g, 95%, red solid) .

LCMS: Rt: 1.244min; MS m / z (ESI): 473 [M + H].

Intermediate Preparation Example 50: Preparation of Intermediate D50

NaH (60 mg, 1.5 mmol) was added portionwise to a solution of compound D7 (230 mg, 1.0 mmol) in DMF (10 mL) at 0 ° C. After the addition was completed, the reaction solution was stirred at 0 ° C for 30 minutes, and then MeOH (48 mg, 1.5 mmol) was added. After the addition, the reaction solution was stirred at 0 ° C for 3 hours. After the reaction was completed, the reaction was quenched with saturated aqueous ammonium chloride solution (50 mL). The resulting mixture was extracted with ethyl acetate (20 mL × 3). The organic phase was washed with water (50 mL) and saturated brine (50 mL × 2) and then dried. Sodium sulfate was dried and filtered, and the filtrate was concentrated under reduced pressure. The crude target product D50 (110 mg, yield: 48.4.1%, yellow liquid) was obtained and used directly in the next step.

LCMS: Rt: 1.208 min; MS m / z (ESI): 228.4 [M + H].

Intermediate Preparation Example 51: Preparation of Intermediate D51

At room temperature, compound D33 (200 mg, 0.54 mmol, compound cyclopropylboronic acid (52 mg, 0.61 mmol), K ₂ CO ₃ (228 mg, 1.662 mmol) and Pd (dppf) Cl ₂ (40 mg, 0.054 mmol) were added to 1, 4In dioxane (5mL) and H ₂ O (0.1mL). After replacing the reaction solution with argon three times, heat to 100 ° C and stir the reaction for 8 hours. The reaction was completed by LCMS. After the reaction solution was cooled to room temperature, water was added ( 20 mL) diluted, the mixture was extracted with dichloromethane (20 mL × 3). The organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was subjected to thin layer chromatography (eluent gradient PE: EtOAc = 1) : 100, R _f = 0.3) purification to obtain the target compound D51 (120 mg, yield: 68%, white solid).

LCMS: Rt: 1.28 min; MS m / z (ESI): 322.2 [M + H].

Intermediate Preparation Example 52: Preparation of Intermediate D52

At 0 ° C, MsCl (2.9 g, 25 mmol) was slowly added to the compound 7-hydroxy-2-azaspiro [3.5] nonane-2-carboxylic acid tert-butyl ester (5.0 g, 21 mmol) in methylene chloride (50 mL ) In the solution, the reaction solution was stirred and reacted at room temperature for 2 hours. After the reaction was completed, saturated aqueous sodium bicarbonate solution (100 mL) was added to the reaction solution to quench the reaction; after the two phases were separated, the aqueous phase was extracted with ethyl acetate (50 mL × 3), and the organic phase was combined with water and saturated brine (100 mL) ) Washed, then dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain the target product D52 (5.0 g, yield: 70%, white solid). The crude product was used directly in the next reaction.

¹ H NMR (400MHz, DMSO-d6) δ: 4.63 (brs, 1H), 3.53-3.50 (m, 4H), 3.16 (s, 3H), 1.82-1.75 (m, 4H), 1.59-1.56 (m, 4H), 1.37 (s, 9H).

Intermediate Preparation Example 53: Preparation of Intermediate D53

At a room temperature, 4-nitropyrazole (4.1 g, 12.8 mmol), D52 (1.6 g, 14.1 mmol), K ₂ CO ₃ (2.7 g, 19.2 mmol), and DMF (50 mL) were sequentially added to the three-necked flask. The reaction solution was stirred at 120 ° C for 2 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and then water (50 mL) and ethyl acetate (50 mL) were added to the reaction solution. After the two phases were separated, the aqueous phase was extracted with ethyl acetate (50 mL × 3). The organic phases were combined and dried and filtered with anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase chromatography (330g column, eluent gradient: water / acetonitrile = 0-100%, time = 100 minutes). To obtain the target product D53 (2.4g, yield: 50%, white solid).

¹ H NMR (400MHz, DMSO) δ: 8.93 (s, 1H), 8.24 (s, 1H), 4.27-4.20 (m, 1H), 3.62-3.51 (m, 4H), 1.95-1.92 (m, 4H) , 1.83-1.76 (m, 2H), 1.63-1.55 (m, 2H), 1.38 (s, 9H).

Intermediate Preparation Example 54: Preparation of Intermediate D54

LiHMDS (18 ml, 18 mmol) was slowly added to a solution of compound D53 (2.4 g, 7.1 mmol) in THF (50 mL) at -60 ° C. After the addition was complete, the reaction solution was stirred at this temperature for 30 minutes. Hexachloroethane (2.1 g, 8.7 mmol) was added to the reaction system in portions, and the reaction was continued to stir for 2 hours. A saturated aqueous ammonium chloride solution (50 mL) was added to the reaction system to quench the reaction, and the resulting mixture was extracted with ethyl acetate (20 mL × 3). The organic phase was washed with saturated brine (50 mL), then dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient: PE: ETOAC = 10: 1-4: 1) to obtain the target product D54 (2.0 g, yield: 70%, white solid).

¹ H NMR (400MHz, DMSO) δ: 8.47 (s, 1H), 4.43-4.37 (m, 1H), 3.64-3.52 (m, 4H), 1.99-1.95 (m, 2H), 1.80-1.73 (m, 2H), 1.68-1.60 (m, 4H), 1.38 (s, 9H).

Intermediate Preparation Example 55: Preparation of Intermediate D55

At a room temperature, D54 (800 mg, 2.2 mmol), iron powder (300 mg, 4.4 mmol), ammonium chloride (600 mg, 10 mmol), and ethanol (10 mL) were added to the single-necked bottle in sequence. The reaction solution was stirred at 80 ° C for 16 hours. After the reaction was completed, the reaction solution was filtered while hot, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D55 (400 mg, yield: 50%, green solid), which was used directly in the next step.

LCMS: Rt: 1.168 min; MS m / z (ESI): 285.3 [M-56 + H].

Intermediate Preparation Example 56: Preparation of Intermediate D56

A mixture of compound D55 (255 mg, 0.84 mmol), compound D12 (300 mg, 0.90 mmol) and ammonium chloride (510 mg, 8.4 mmol) in ethanol (30 mL) was heated to 80 ° C. and stirred for 72 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and then concentrated under reduced pressure, and the residue was purified by column chromatography (eluent gradient DCM / MeOH = 50/1 to 10/1) to obtain the target compound D56 (600 mg, Yield:> 100%, green solid) directly used in the next step.

LCMS: Rt: 1.075 min; MS m / z (ESI): 620.4 [M + H].

Intermediate Preparation Example 57: Preparation of Intermediate D57

At room temperature, compound 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester (2.0 g, 9 mmol) and compound 2-aminophenyl-dimethylphosphine oxide (1.9 g, 10.8) were added to a 100 mL three-necked flask, respectively. mmol), DIEA (3.5 g, 27 mmol) and DMF (20 mL). The mixture was heated to 25 ° C and stirred for 16 hours. After the reaction was completed, the reaction mixture was poured into water (50 mL). The mixture was extracted with ethyl acetate (50 mL × 3), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient DCM / MeOH = 80: 1-40: 1) to obtain the target product D57 (1.5 g, yield: 45%, yellow solid).

¹ H NMR (400MHz, DMSO) δ: 11.0 (s, 1H), 8.80 (s, 1H), 7.88-7.84 (m, 1H), 7.71-7.59 (m, 2H), 7.37-7.33 (m, 1H) , 4.37 (q, J = 6.8Hz, 2H), 1.74 (s, 3H), 1.71 (s, 3H), 1.36 (t, J = 6.8Hz, 3H)

Intermediate Preparation Example 58: Preparation of Intermediate D58

At room temperature, compound D15 (611.7 mg, 2 mmol), MeOH (5 mL) and Pd / C (61.2 mg, 10%) were added in sequence in a 100 mL single-necked bottle. The reaction system was stirred at room temperature for 16 hours under a hydrogen atmosphere. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd / C, and the filtrate was concentrated under reduced pressure to obtain the crude target product D58 (551.7 mg, yield: 100%). The crude product was used directly in the next reaction.

MS / m / z (ESI): 276.9 [M + H].

Intermediate Preparation Example 59: Preparation of Intermediate D59

Compound D50 (116 mg, 0.51 mmol) was dissolved in MeOH (20 mL) at room temperature, and then Pd / C (20 mg) was added; after the addition, the reaction system was replaced with hydrogen three times, and then the system was stirred at room temperature for 2 h. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd / C, and the filtrate was concentrated under reduced pressure to obtain the target crude product D59 (100.6 mg, yield: 100%). The crude product was used directly in the next reaction.

MS / m / z (ESI): 198.2 [M + H].

Intermediate Preparation Example 60: Preparation of Intermediate D60

At room temperature, D4 (1.0 g, 3.2 mmol) was dissolved in a solution of hydrogen chloride gas in dioxane (6N, 10 mL). After the solid was completely dissolved, the reaction solution was stirred at room temperature for 2 hours. Thin layer chromatography (petroleum ether: ethyl acetate = 1: 1, R _f = 0.2) showed that the starting material disappeared and the product was formed. The reaction solution was concentrated under reduced pressure to obtain the target compound D60 (1.1 g, crude product, white solid). The crude product was used directly in the next reaction.

Intermediate Preparation Example 61: Preparation of Intermediate D61

At room temperature, add D60 (500mg, 2.36mmol) and 1,2-dichloroethane (10mL) to a 100mL single-necked bottle, and then add aqueous formaldehyde (3mL); the mixture was stirred at room temperature for 1 hour and triacetic acid was added Sodium borohydride (995 mg, 4.72 mmol) and 5 drops of acetic acid. After the addition was complete, the reaction mixture continued to react at room temperature for 10 hours. LCMS detected that the reaction was complete, the reaction solution was diluted with water (20 mL), the mixture was extracted with dichloromethane (20 mL × 3); the organic phases were combined, dried and filtered with anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure, and the residue was prepared by thin layer chromatography The analytical method (eluent gradient: dichloromethane / methanol = 20: 1, R _f = 0.2) was purified to obtain the target compound D61 (200 mg, Yield: 37.8%, white solid).

LCMS: Rt: 0.32 min; MS m / z (ESI): 225.3 [M + H].

Intermediate Preparation Example 62: Preparation of Intermediate D62

At room temperature, D61 (200 mg, 0.89 mmol) was dissolved in ethanol (5 mL) and palladium / carbon catalyst (20 mg) was added, and the reaction system was heated to 60 ° C. under a hydrogen atmosphere and stirred for 2 hours. The reaction was checked by LCMS, the mixture was cooled and filtered, and the filtrate was concentrated under reduced pressure to obtain the target compound D62 (150 mg, Yield: 86%, white solid).

LCMS: Rt: 0.32 min; MS m / z (ESI): 195.3 [M + H].

Intermediate Preparation Example 63: Preparation of Intermediate D63

At room temperature, (R) -tetrahydrofuran-3-yl methyl methanesulfonate (2.0g, 12.03mmol), 4-nitropyrazole (1.5g, 13.24mmol) and N, N -Dimethylformamide (20 mL), potassium carbonate (4.99 g, 36.10 mmol) was added, and the reaction solution was heated to 120 ° C. and stirred overnight. The reaction was completed by LCMS. After the reaction solution was cooled to room temperature, water (30 mL) and ethyl acetate (30 mL) were added. After the two phases were separated, the organic phase was washed with water (50 mL), dried and filtered with anhydrous sodium sulfate, and the filtrate was decompressed. Concentrate to obtain the target crude product D63 (2.5g, yield: 70%, white solid). The crude product was used directly in the next reaction.

LCMS: Rt: 1.253 min; MS m / z (ESI): 184.4 [M + H].

Intermediate Preparation Example 64: Preparation of Intermediate D64

D63 (300.0 mg, 1.64 mmol), 1,2-dichloroethane (425.6 mg, 1.80 mmol) and tetrahydrofuran (5 mL) were added to a 25 mL three-necked bottle, and the reaction system was cooled to -60 ° C under a nitrogen atmosphere. Then lithium bistrimethylsilylamine (1.64 mL, 1.64 mmol) was slowly added dropwise. After the dropwise addition, the reaction mixture was further stirred at -60 ° C for 2 hours. LCMS detected that the reaction was complete. To the reaction solution was added saturated aqueous ammonium chloride solution (25 mL) to quench the reaction. The mixture was extracted with ethyl acetate (50 mL). The organic phase was dried and filtered with anhydrous Na ₂ SO ₄ and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent gradient: petroleum ether: ethyl acetate = 10: 1, Rf = 0.1) to obtain the target product D64 (110 mg, yield: 31.0%, white solid).

LCMS: Rt: 2.463 min; MS m / z (ESI): 217.4 [M + H].

Intermediate Preparation Example 65: Preparation of Intermediate D65

At room temperature, dissolve D64 (110.0 mg, 0.51 mmol) and methylboronic acid (303.0 mg, 5.1 mmol) in dioxane / water (2 mL / 0.5 mL), then add sodium carbonate (265.0 mg, 2.5 mmol) And [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride (36.6 mg, 0.05 mmol). The reaction system was protected with nitrogen and heated to 100 ° C and stirred overnight. After the reaction was completed, the reaction mixture was cooled and filtered. The filtrate was diluted with water (20 mL) and ethyl acetate (20 mL). After the two phases were separated, the aqueous phase was extracted with ethyl acetate (20 mL). The ethyl acetate phases were combined and dried over anhydrous sodium sulfate. It was dried and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent gradient: petroleum ether: ethyl acetate = 10: 1, Rf = 0.2) to obtain the target product D65 (60.0 mg, yield : 60%, brown solid).

Intermediate Preparation Example 66: Preparation of Intermediate D66

At room temperature, D65 (60.0 mg, 0.3 mmol) was dissolved in tetrahydrofuran (5 mL), and then a palladium carbon catalyst (30.0 mg, 50%) was added. The reaction system was stirred at room temperature for 2h under a hydrogen atmosphere. The reaction was detected by LCMS, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the target product D66 (50.0 mg, yield: 100%, yellow solid).

LCMS: Rt: 0.273 min; MS m / z (ESI): 168.5 [M + H].

Intermediate Preparation Example 67: Preparation of Intermediate D67

At room temperature, to 5-chloro-4-nitro-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazole (500 mg, 2.16 mmol) in N, N-dimethylformamide ( 4.3mL) solution was added sodium cyanide (212mg, 4.32mmol), the reaction system was heated to 80 ℃ under argon protection and stirred for 7 hours; after the reaction was complete, the reaction system was cooled to room temperature, and then diluted with water (20mL) The mixture was extracted with ethyl acetate (10mL × 3). The combined organic phases were washed with water (30 mL), the organic solution was dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D67 (400 mg, Yield 60%, yellow solid).

LCMS: Rt: 1.635 min; MS m / z (ESI): 461 [M + H ₂ OH].

Intermediate Preparation Example 68: Preparation of Intermediate D68

At room temperature, palladium carbon (10 wt%, 100 mg) was added to a solution of D67 (200 mg, 0.9 mmol) in tetrahydrofuran (5 mL), and the reaction system was stirred at room temperature for 6 hours under a hydrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D68 (150 mg, Yield: 80%, yellow solid).

LCMS: Rt: 1.254min; MS m / z (ESI): 402 [2M + NH 4].

Intermediate Preparation Example 69: Preparation of Intermediate D69

At room temperature, D33 (200 mg, 0.55 mmol), 4,4,5,5-tetramethyl-2- (isopropyl-1-en-2-yl) -1,3,2-dioxaborate (93 mg, 0.55 mmol), potassium carbonate (227 mg, 1.65 mmol) and 1,1′-bisdiphenylphosphinoferrocene palladium dichloride (40 mg, 0.05 mmol) were dissolved in dioxane (10 mL). The reaction system was heated to 100 ° C under the protection of nitrogen, and the reaction was stirred for 8 hours. The reaction was completed by LCMS. After cooling, the reaction solution was diluted with water (20 mL). The mixture was extracted with dichloromethane (30 mL × 5). The organic phase was dried and filtered with sodium sulfate. The filtrate was concentrated under reduced pressure. Removal gradient: dichloromethane / methanol = 20/1, Rf = 0.4) purification to obtain the target compound D69 (110 mg, yield: 62.5%, white solid).

LCMS: Rt: 1.653 min; MS m / z (ESI): 322.2 [M + H].

Intermediate Preparation Example 70: Preparation of Intermediate D70

At room temperature, D69 (100 mg, 0.3 mmol) was dissolved in ethyl acetate (10 mL), and then a palladium / carbon catalyst (20 mg) was added. The reaction system was stirred under a hydrogen atmosphere at room temperature for 2 hours, and the reaction was completed by LCMS. The reaction mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by thin-layer preparative chromatography (eluent gradient: dichloromethane: methanol = 20: 1, Rf = 0.6) to obtain the target compound D70 (30 mg, yield: 30%, white solid).

LCMS: Rt: 1.49 min; MS m / z (ESI): 324.2 [M + H].

Intermediate Preparation Example 71: Preparation of Intermediate D71

At room temperature, D2 (640 mg, 2.16 mmol) was dissolved in methanol (30 mL), and then a palladium-carbon catalyst (120 mg) was added. The reaction system was stirred at room temperature under a hydrogen atmosphere for 16 hours. The reaction was detected by LCMS, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the target crude product D71 (550 mg, yield: 95.6%, yellow oily liquid). The crude product was used directly in the next reaction.

LCMS: Rt: 0.933 min; MS m / z (ESI): 267.4 [M + H].

Intermediate Preparation Example 72: Preparation of Intermediate D72

At room temperature, dissolve D71 (225 mg, 0.834 mmol) and D33 (150 mg, 0.417 mmol) in ethanol (30 mL), add ammonium chloride (225 mg, 4.2 mmol) to the reaction system; then heat the reaction solution to 80 ° C Stir for 16 hours. After the reaction, the reaction solution was cooled and filtered, the filter cake was washed with ethanol (10 mL × 2), the filtrate was concentrated under reduced pressure, and the residue was subjected to preparative thin layer chromatography (eluent gradient: dichloromethane: methanol = 20: 1) Purification to obtain the target intermediate D72 (55 mg, yield: 22.3%, yellow solid).

LCMS: Rt: 1.425 min; MS m / z (ESI): 592.4 [M + 3].

Intermediate Preparation Example 73: Preparation of Intermediate D73

At room temperature, D72 (55 mg, 0.093 mmol) was dissolved in a solution of hydrogen chloride in dioxane (4M, 10 mL), and the reaction was carried out at room temperature for 2 hours. LCMS detected the reaction was complete, and the reaction solution was concentrated under reduced pressure to obtain the hydrochloride salt of the target intermediate D73 (50 mg, yield: 100%, yellow solid).

LCMS: Rt: 0.717 min; MS m / z (ESI): 490.2 [M + H].

Intermediate Preparation Example 74: Preparation of Intermediate D74

At room temperature, D3 (1.5g, 5mmol), iron powder (1.4g, 25mmol), ammonium chloride (2.16g, 40mmol) was added to a mixture of ethanol (28mL) and water (7mL), the reaction solution was in argon Heat to 80 ° C under air and stir for 1 hour. After the reaction was completed, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the target crude product D74 (1.3 g, yield: 96%), character: red solid.

LCMS: Rt: 1.395 min; MS m / z (ESI): 245.2 [M-56 + H].

Intermediate Preparation Example 75: Preparation of Intermediate D75

At room temperature, D74 (1.16 g, 3.7 mmol), D12 (1 g, 3.7 mmol), and ammonium chloride (951 mg, 18.5 mmol) were dissolved in ethanol (30 mL), and the reaction was stirred at 80 ° C. for 12 hours. After the reaction was completed, the reaction mixture was poured into water (100 mL), the mixture was extracted with dichloromethane / methanol (10/1, 30 mL × 3), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent gradient: dichloromethane: methanol = 20: 1, Rf = 0.7) to obtain the target product D75 (1.2g, yield: 54%), properties: white solid .

LCMS: Rt: 1.539 min; MS m / z (ESI): 580.4 [M + H].

Intermediate Preparation Example 76: Preparation of Intermediate D76

At room temperature, a solution of D75 (220 mg, 0.38 mmol) and 4M hydrogen chloride dioxane (0.95 mL, 3.8 mmol) was dissolved in absolute ethanol (4 mL), and the reaction solution was stirred at room temperature for 2 hours. LCMS detected that the reaction was complete, and the reaction mixture was concentrated under reduced pressure to obtain the crude target product D76 (182 mg, yield: 95%), character: light yellow solid.

LCMS: Rt: 1.62 min; MS m / z (ESI): 480.1 [M + H].

Intermediate Preparation Example 77: Preparation of Intermediate D77

At room temperature, 7- (4-nitro-1H-pyrazol-1-yl) -2-azaspiro [3.5] nonane (156 mg, 0.66 mmol), 1-bromo-2-methoxyethane (109 mg, 0.79 mmol) and potassium carbonate (96 mg, 0.79 mmol) were added to acetonitrile (5 mL), and then the reaction solution was heated to 80 ° C. and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D77 (120 mg, yield: 91%), character: yellow solid.

LCMS: Rt: 0.716 min; MS m / z (ESI): 295.2 [M + H].

Intermediate Preparation Example 78: Preparation of Intermediate D78

At room temperature, wet palladium carbon (12 mg) was added to a solution of D77 (120 mg, 041 mmol) in ethanol (5 mL), and the reaction system was heated to 30 ° C. under a hydrogen atmosphere and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D78 (100 mg, yield: 93%), properties: white solid

LCMS: Rt: 0.312 min; MS m / z (ESI): 265.4 [M + H].

Intermediate Preparation Example 79: Preparation of Intermediate D79

At room temperature, trifluoroacetic acid (2 mL) was added to a solution of D53 (300 mg, 0.86 mmol) in methylene chloride (8 mL), and the reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the target compound D79 (312 mg, yield: 100%), character: white solid, which was directly used in the next step.

LCMS: Rt: 0.727 min; MS m / z (ESI): 237.2 [M + H].

Intermediate Preparation Example 80: Preparation of Intermediate D80

To a solution of D79 (156 mg, 0.66 mmol) and formaldehyde in water (198 mg, 37% aqueous solution, 10.0 mmol) in methanol (5 mL) was added triethylamine (66.7 mg, 0.66 mmol) at room temperature. After stirring for 5 hours, boron triacetate was added Sodium hydride (699.6 mg, 3.3 mmol), and the reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by thin-layer preparative chromatography (eluent gradient: dichloromethane / methanol = 10/1) to obtain the target compound D80 (100 mg, yield: 60%) , Properties: White solid

LCMS: Rt: 0.808 min; MS m / z (ESI): 321.5 [M + H].

Intermediate Preparation Example 81: Preparation of Intermediate D81

At room temperature, wet palladium carbon (8.0 mg) was added to a solution of D80 (80 mg, 0.32 mmol) in ethanol (5 mL), and the reaction system was heated to 30 ° C. under a hydrogen atmosphere and stirred for 4 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the target compound D81 (56 mg, yield: 80%), properties: yellow solid

LCMS: Rt: 0.291 min; MS m / z (ESI): 221.2 [M + H].

Intermediate Preparation Example 82: Preparation of Intermediate D82

At room temperature, D77 (1066 mg, 6.16 mmol) and diisopropylethylamine (751 mg, 6.16 mmol) were dissolved in acetonitrile (10 mL), and then the reaction solution was heated to 80 ° C. and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D82 (560 mg, yield: 65%), character: yellow solid.

LCMS: Rt: 0.885 min; MS m / z (ESI): 283.3 [M + H].

Intermediate Preparation Example 83: Preparation of Intermediate D83

At room temperature, wet palladium carbon (56 mg) was added to a solution of D82 (560 mg, 1.98 mmol) in ethanol (10 mL), and the reaction system was heated to 30 ° C. under a hydrogen atmosphere and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D83 (360 mg, yield: 93%), properties: yellow solid.

LCMS: Rt: 0.259 min; MS m / z (ESI): 253.1 [M + H].

Intermediate Preparation Example 84: Preparation of Intermediate D84

At room temperature, dissolve tert-butyl 5-oxocyclopentyl [c] pyrrolidine-2 (1H) -carboxylate (10g, 44.4mmol) in ethanol (200mL), and then cool to 0 ° C. Sodium borohydride (5.1 g, 133.2 mmol) was slowly added to the reaction system, and the reaction was allowed to react overnight at room temperature. The reaction was completed by TLC, water (100 mL) was slowly added dropwise to the reaction solution, the compound was extracted with ethyl acetate (50 mL × 4), the organic phase was combined and washed with saturated brine (50 mL × 3), and then dried and filtered over anhydrous sodium sulfate The filtrate was concentrated under reduced pressure to obtain the crude target compound D84 (9g, yield: 95%), properties: yellow liquid.

¹ H NMR (400 MHz, CDCl ₃ ) δ 4.28-4.21 (m, 1H), 3.43-3.41 (m, 2H), 3.29-3.26 (m 2H), 2.53 (br, 2H), 2.10-2.06 (m, 2H), 1.48-1.43 (m, 1H), 1.42-1.40 (m, 1H), 1.39 (s, 9H).

Intermediate Preparation Example 85: Preparation of Intermediate D85

At room temperature, D84 (10.0g, 44.4mmol) was dissolved in dichloromethane (200mL), then triethylamine (9.0g, 88.8mmol) was added, the reaction solution was cooled to -5 ℃ and then slowly added to the system Methanesulfonyl chloride (10.2g, 88.8mmol), control the temperature of the system not to exceed 5 ° C, and after the dropwise addition, let the reaction stir at room temperature overnight. LCMS detected the reaction was complete, added saturated sodium bicarbonate solution (100 mL), and stirred for 10 minutes, the mixture was extracted with dichloromethane (50 mL X 3), the combined organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain The crude target product D85 (1.5g, yield: 70%), properties: yellow solid. The crude product was used directly in the next reaction.

¹ H NMR (400 MHz, CDCl ₃ ): δ 5.17-5.04 (m, 1H), 3.56-3.53 (m, 2H), 3.36-3.33 (m, 2H), 3.09-2.96 (m, 5H), 2.67 ( br, 2H), 2.35-2.32 (m, 2H), 1.88-1.85 (m, 2H), 1.46 (s, 9H).

Intermediate Preparation Example 86: Preparation of Intermediate D86

At room temperature, D85 (5.2 g, 46 mmol), 4-nitro-1H-pyrazole (14 g, 46 mmol), potassium carbonate (12.7 g, 92 mmol) were added to N, N-dimethylformamide (100 mL), The reaction solution was heated to 120 ° C and stirred overnight. The reaction was completed by LCMS, water (100 mL) was added to the reaction solution, the mixture was extracted with ethyl acetate (50 mL × 3), the combined organic phase was washed with saturated brine (50 mL × 5), and the organic phase was dried and filtered over anhydrous sodium sulfate The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent gradient: petroleum ether: ethyl acetate = 10: 1-ethyl acetate) to obtain the target compound D86 (6g, yield: 40%) ), Character: yellow oil.

LCMS: Rt: 1.231 min; MS m / z (ESI): 267 [M-55].

Intermediate Preparation Example 87: Preparation of Intermediate D87

At room temperature, D86 (1.5 g, 4.6 mmol) was dissolved in methanol (10 mL), then a solution of hydrogen chloride in 1,4-dioxane (10 mL, 4N) was added, and the reaction was stirred at room temperature for 2 hours. LCMS detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure to obtain the crude target compound D87 (1.1 g, yield: 100%), character: white solid. The crude product was used directly in the next reaction.

LCMS: Rt: 0.599 min; MS m / z (ESI): 223 [M + H].

Intermediate Preparation Example 88: Preparation of Intermediate D88

At room temperature, D87 (1.1 g, 4.9 mmol) was dissolved in methanol (20 mL), and formaldehyde (1.5 g, 15 mL, 37%) was added, and the reaction solution was stirred at room temperature overnight. LCMS showed that the reaction was complete, sodium triacetoxyborohydride (3.2 g, 15 mmol) was added, and the reaction solution was stirred at room temperature for 15 minutes. LCMS showed that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (eluent gradient: petroleum ether: ethyl acetate = 10: 1; 1,2-dichloromethane: methanol = 10: 1 ) Purification to obtain the target compound D88 (800 mg, yield 68%), properties: yellow oil.

LCMS: Rt: 0476 min; MS m / z (ESI): 236 [M + H].

Intermediate Preparation Example 89: Preparation of Intermediate D89

At room temperature, D88 (800 mg, 3.4 mmol) was dissolved in ethanol (20 mL), a palladium-carbon catalyst (200 mg) was added, the system was replaced with hydrogen three times, and the reaction was stirred at room temperature under a hydrogen atmosphere for 3 hours. LCMS showed that the reaction was complete, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D89 (500 mg, yield: 66%), character: yellow oil. The crude product was used directly in the next reaction.

LCMS: Rt: 0.233 min; MS m / z (ESI): 207 [M + H].

Intermediate Preparation Example 90: Preparation of Intermediate D90

At room temperature, D124 (1.2 g, 2.2 mmol) and Lawesson's reagent (1.78 g, 4.4 mmol) were dissolved in toluene (50 mL), and the reaction solution was stirred at 120 ° C for 3 hours. The product was detected by LCMS, the reaction solution was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent gradient: methanol: dichloromethane = 0-10%) to obtain the target product D90 (350 mg, Yield: 30%), character: light yellow solid.

LCMS: Rt: 1.84 min; MS m / z (ESI): 562.5 [M + H].

Intermediate Preparation Example 91: Preparation of Intermediate D91

At room temperature, D90 (350 mg, 0.62 mmol) was dissolved in 4M hydrogen chloride dioxane (1.55 mL, 6.2 mmol) solution, and the reaction solution was stirred at room temperature for 2 hours. After LCMS detected that the reaction was complete, the reaction mixture was concentrated under reduced pressure to obtain the crude target product D91 (310 mg, yield: 98%), character: light yellow solid.

LCMS: Rt: 1.15 min; MS m / z (ESI): 462.3 [M + H].

Intermediate Preparation Example 92: Preparation of Intermediate D92

At room temperature, dissolve tert-butyl 6-hydroxy-2-azaspiro [3.3] heptane-2-carboxylate (3.0 g, 14.1 mmol) in 1,2-dichloromethane (30 mL) and add Triethylamine (2.9g, 28.2mmol). After the reaction solution was cooled to 0 ° C, methanesulfonyl chloride (1.6g, 14.1mmol) was slowly added to the system; the reaction solution was warmed to room temperature and stirred overnight. TLC detected the completion of the reaction. Water (100 mL) was added to the reaction solution and stirred for 15 minutes. The mixture was extracted with dichloromethane (50 mL × 4). The organic phases were combined and washed with saturated brine (50 mL × 3), and then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to obtain the crude target compound D92 (2.8 g, yield: 70%), character: yellow solid. The crude product was used directly in the next reaction.

LCMS: MS / m (zI): N / A.

Intermediate Preparation Example 93: Preparation of Intermediate D93

At room temperature, D92 (3.5g, 12mmol), 4-nitro-1H-pyrazole (1.4g, 12mmol), potassium carbonate (3.3g, 24mmol) was added to N, N-dimethylformamide (50mL) , The reaction solution was heated to 120 ℃ reaction overnight. LCMS detected that the reaction was complete. Water (100 mL) was added to the reaction solution, the mixture was extracted with ethyl acetate (50 mL × 3), the organic phase was combined and washed with saturated brine (50 mL × 5), and then dried and filtered with anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure to obtain crude target compound D93 (2.0 g, yield: 70%), character: yellow solid. The crude product was used directly in the next reaction.

LCMS: Rt: 1.585 min; MS m / z (ESI): 309 [M + H].

Intermediate Preparation Example 94: Preparation of Intermediate D94

Dissolve tert-butyl 6- (4-nitro-1H-pyrazol-1-yl) -2-azaspiro [3.3] heptane-2-carboxylate (500g, 1.6mmol) in methanol at room temperature To (10 mL), a solution of hydrochloric acid gas in 1,4-dioxane (10 mL, 4N) was added, and the reaction was stirred at room temperature for 2 hours. LCMS detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure to obtain the crude target compound D94 (430 mg, yield: 100%), character: white solid. The crude product was used directly in the next reaction.

LCMS: Rt: 0.353 min; MS m / z (ESI): 209 [M + H].

Intermediate Preparation Example 95: Preparation of Intermediate D95

At room temperature, D94 (430 mg, 2.1 mmol) was dissolved in methanol (20 mL), and formaldehyde (511 g, 6.3 ml, 37%) was added and stirred at room temperature overnight. After LCMS showed that the reaction was complete, sodium triacetoxyborohydride (1.4 g, 6.3 mmol) was added, and the reaction was continued to be stirred at room temperature for 15 minutes. LCMS showed the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent gradient: petroleum ether: ethyl acetate = 10: 1, 2-dichloromethane: methanol = 10: 1) , To obtain the target compound D95 (250mg, yield: 55%), properties: yellow oil.

LCMS: Rt: 0.463 min; MS m / z (ESI): 223 [M + H].

Intermediate Preparation Example 96: Preparation of Intermediate D96

At room temperature, D95 (200 mg, 0.9 mmol) was dissolved in ethanol (10 mL), a palladium-carbon catalyst (100 mg) was added, the system was replaced with hydrogen three times, and the reaction was stirred at room temperature under a hydrogen atmosphere for 3 hours. LCMS showed that the reaction was complete, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D96 (150 mg, yield: 80%). Traits: yellow oil. The crude product was used directly in the next reaction.

LCMS: Rt: 0312 min; MS m / z (ESI): 193 [M + H].

Intermediate Preparation Example 97: Preparation of Intermediate D97

At room temperature, 3- (benzyloxy) cyclobutanone (3.52g, 20.0mmol) and 1-methylpiperazine (2.20g, 22.0mmol) were dissolved in 1,2-dichloroethane (300mL), Then sodium borohydride triacetate (8.46 g, 40.0 mmol) was added in portions; the reaction was stirred at room temperature for 3 hours. The reaction was completed by LCMS. A saturated sodium bicarbonate aqueous solution (100 mL) was added to the system. The compound was extracted with ethyl acetate (50 mL × 3). The organic phase was combined and washed with saturated brine (100 mL), then dried and filtered with anhydrous sodium sulfate, and the filtrate It was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent gradient: methanol / dichloromethane = 0 to 10%) to obtain the target product D97 (4.2g, yield: 81%), properties: none Color oil.

LCMS: Rt: 1.75 min; MS m / z (ESI): 261.3 [M + H].

Intermediate Preparation Example 98: Preparation of Intermediate D98

At room temperature, D97 (2.0 g, 7.7 mmol) was dissolved in trifluoroacetic acid (9 mL), and the reaction solution was stirred at 80 ° C. for 12 hours. LCMS detected that the reaction was complete, and the reaction mixture was concentrated under reduced pressure to obtain the crude target product D98 (1.31 g), character: light yellow liquid.

LCMS: Rt: 0.31 min; MS m / z (ESI): 171.4 [M + H].

Intermediate Preparation Example 99: Preparation of Intermediate D99

Under ice bath, dissolve D98 (1.31 g, 7.7 mmol) in dichloromethane (50 mL), add triethylamine (3.85 g, 38.5 mmol) first, and then add methanesulfonyl chloride (0.9 mL, 11.55 mmol). The reaction solution was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate solution (100 mL) was added to the system, and the compound was extracted with ethyl acetate (20 mL * 3). The combined organic phases were washed with saturated brine (100 mL), then dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure To get the crude target product D99 (1.9g), character: light yellow liquid.

LCMS: Rt: 0.43 min; MS m / z (ESI): 249.3 [M + H].

Intermediate Preparation Example 100: Preparation of Intermediate D100

At room temperature, D99 (1.9 g, 7.7 mmol), 4-nitro-1H-pyrazole (1.31 g, 11.55 mmol) and potassium carbonate (3.20 g, 23.1 mmol) were added to N, N-dimethylformamide ( In 20 mL), the reaction solution was stirred at 120 ° C for 12 hours. LCMS detected product formation, the reaction solution was diluted with water (50 mL), the compound was extracted with ethyl acetate (40 mL × 2), the organic phase was combined and washed with saturated brine (100 mL × 2), then dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure , The residue was purified by silica gel column chromatography (eluent gradient: methanol / dichloromethane = 0 ~ 10%) to obtain the target product D100 (120mg, three-step yield: 6%), properties: light yellow solid .

LCMS: Rt: 0.43 min; MS m / z (ESI): 266.4 [M + H].

Intermediate Preparation Example 101: Preparation of Intermediate D101

At room temperature, D100 (120 mg, 0.45 mmol) and palladium carbon catalyst (60 mg) were added to absolute ethanol / tetrahydrofuran (10 mL / 1 mL). After the reaction solution was replaced with hydrogen three times, the reaction was stirred at room temperature for 1 hour. LCMS detected the reaction was complete, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target product D101 (106 mg, yield: 98%), properties: light yellow solid.

LCMS: Rt: 0.43 min; MS m / z (ESI): 236.3 [M + H].

Intermediate Preparation Example 102: Preparation of Intermediate D102

D18 (153 mg, 0.63 mmol) and D12 (200 mg, 0.63 mmol) were dissolved in isopropanol (10 mL), then trifluoroacetic acid (1 mL) was added, and the reaction liquid was reacted at 100 ° C. for 10 hours. LCMS detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was purified by thin-layer preparative chromatography (eluent gradient: dichloromethane / methanol = 10: 1, Rf = 0.3) to obtain the target compound D102 (190 mg , Yield: 73.9%), character: yellow solid.

LCMS: Rt: 1.261 min; (0.1% FA-H ₂ O): (0.1% FA-ACN) = 90: 10; MS m / z (ESI): 407.3 [M + H].

Intermediate Preparation Example 103: Preparation of Intermediate D103

D102 (100 mg, 0.24 mmol) was dissolved in toluene (10 mL), dichlorosulfoxide (0.5 mL) was added, and the reaction solution was heated to 70 ° C. for 4 hours. LCMS detected that the reaction was complete. The reaction solution was concentrated under reduced pressure. The residue was diluted with water (20 mL) and then extracted with dichloromethane (20 mL × 3). The combined organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain the target compound D103 (80 mg, yield: 76.9%), character: brown solid.

LCMS: Rt: 1.421 min; (0.1% FA-H ₂ O): (0.1% FA-ACN) = 90: 10; MS m / z (ESI): 425.2 [M + H].

Intermediate Preparation Example 104: Preparation of Intermediate D104

At room temperature, D71 (500 mg, 0.85 mmol) and Lawson's reagent (688 mg, 1.70 mmol) were dissolved in toluene (10 mL), and the reaction solution was stirred at 120 ° C for 3 hours. The product was detected by LCMS, the reaction solution was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent gradient methanol / dichloromethane = 0 to 10%) to obtain the target product D104 (330 mg, yield Rate: 50%), traits: light yellow solid.

LCMS: Rt: 1.85 min; MS m / z (ESI): 608.2 [M + H].

Intermediate Preparation Example 105: Preparation of Intermediate D105

At room temperature, D104 (330 mg, 0.54 mmol) and trifluoroacetic acid (2.0 mL) were dissolved in dichloromethane (5 mL), and the reaction solution was stirred at room temperature for 2 hours. LCMS detected that the reaction was complete, and the reaction mixture was concentrated under reduced pressure to obtain the crude target product D105 (210 mg, yield: 76%), character: light yellow solid.

LCMS: Rt: 1.16 min; MS m / z (ESI): 508.2 [M + H].

Intermediate Preparation Example 106: Preparation of Intermediate D106

At room temperature, tert-butyl 3-hydroxypiperidine-1-carboxylate (10 g, 49.7 mmol) and triethylamine (15 g, 149 mmol) were added to dichloromethane (150 mL). The mixture was replaced with argon three times and then cooled to -20 ° C. Methanesulfonyl chloride (11.38g, 99.4mmol) was slowly added dropwise to the system. After the addition, the reaction was stirred at room temperature for 4 hours. After the reaction was completed, the reaction mixture was poured into saturated aqueous sodium bicarbonate solution (300 mL), the mixture was extracted with ethyl acetate (100 mL × 3), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain the crude target product D106 (12g, yield: 87%), properties: red liquid.

LCMS: Rt: 1.615 min; MS m / z (ESI): 224.1 [M-56 + H].

Intermediate Preparation Example 107: Preparation of Intermediate D107

At room temperature, dissolve D106 (12g, 42.96mmol), 4-nitro-1H-pyrazole (9.71g, 85.91mmol), potassium carbonate (17.81g, 128.88mmol) in N'N-dimethylformamide (160 mL), the reaction was stirred at 110 ° C. for 12 hours. After the reaction was completed, the reaction mixture was poured into water (500 mL), the mixture was extracted with ethyl acetate (100 mL × 3), the organic phase was washed with saturated aqueous sodium carbonate solution (300 mL × 8), and then dried and filtered with anhydrous sodium sulfate, and the filtrate It was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent gradient: petroleum ether: ethyl acetate = 1: 1, Rf = 0.4) to obtain the target product D107 (7.5 g, yield: 59%) , Properties: yellow liquid.

LCMS: Rt: 1.772 min; MS m / z (ESI): 297.1 [M + H].

Intermediate Preparation Example 108: Preparation of Intermediate D108

At room temperature, D107 (200 mg, 0.67 mmol) and palladium-carbon catalyst (100 mg) were added to methanol (10 mL). The reaction system was replaced with hydrogen three times and stirred at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the target product D108 (170 mg, yield: 95%), character: yellow liquid.

LCMS: Rt: 1.007 min; MS m / z (ESI): 267.2 [M + H].

Intermediate Preparation Example 109: Preparation of Intermediate D109

At room temperature, D108 (170 mg, 0.64 mmol), D12 (202 mg, 0.64 mmol) and ammonium chloride (171 mg, 3.19 mmol) were dissolved in ethanol (8 mL), and the reaction was stirred at 80 ° C. for 12 hours. After the reaction was complete, the reaction mixture was poured into water (40 mL). The mixture was extracted with dichloromethane / methanol (10/1, 30 mL × 3), the organic phase was dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent gradient: dichloromethane: methanol = 20: 1, Rf = 0.5) to obtain the target product D109 (110 mg, yield: 32%), character: white liquid.

LCMS: Rt: 1.488 min; MS m / z (ESI): 546.2 [M + H].

Intermediate Preparation Example 110: Preparation of Intermediate D110

At room temperature, D109 (110 mg, 0.2 mmol) was dissolved in a mixed solution of hydrogen chloride in dioxane (4M, 0.6 mL) and methanol (1 mL). After the reaction was stirred at room temperature for 1 hour, the mixture was concentrated under reduced pressure. The crude target compound D110 (80 mg, yield: 89%) was obtained. Properties: colorless liquid.

LCMS: Rt: 0.804 min; MS m / z (ESI): 446.1 [M + H].

Intermediate Preparation Example 111: Preparation of Intermediate D111

At room temperature, 7- (4-nitro-1H-pyrazol-1-yl) -2-azaspiro [3.5] nonane (100 mg, 0.42 mmol), 1,1-difluoro-2-iodoethyl Alkane (160 mg, 0.84 mmol) and diisopropylethylamine (102 mg, 0.84 mmol) were dissolved in acetonitrile (5 mL), and then the reaction solution was heated to 80 ° C. and stirred for 16 hours. After the reaction was completed, the reaction liquid was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D111 (100 mg, yield: 91%), properties: black liquid

LCMS: Rt: 0.823 min; MS m / z (ESI): 301.3 [M + H].

Intermediate Preparation Example 112: Preparation of Intermediate D112

At room temperature, wet palladium carbon (12 mg) was added to a solution of D111 (45 mg, 015 mmol) in ethanol (5 mL), and the reaction system was heated to 30 ° C. under a hydrogen atmosphere and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D112 (30 mg, yield: 75%), character: black solid.

LCMS: Rt: 1.028 min; MS m / z (ESI): 271.2 [M + H].

Intermediate Preparation Example 113: Preparation of Intermediate D113

At room temperature, wet palladium carbon (50 mg) was added to a solution of D53 (0.5 g, 1.49 mmol) in ethanol (30 mL), and the reaction system was heated to 30 ° C. under a hydrogen atmosphere and stirred for 6 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D113 (1.8 g, yield: 100%). Properties: white solid.

LCMS: Rt: 0.858 min; MS m / z (ESI): 307.1 [M + H].

Intermediate Preparation Example 114: Preparation of Intermediate D114

At room temperature, dissolve D12 (1.65g, 5.2mmol), D113 (1.6g, 5.2mol) and ammonium chloride (2.76g, 5.2mmol) in ethanol (5mL), then heat the reaction solution to 80 ° C and stir overnight . After the reaction was completed, the reaction solution was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent gradient: methylene chloride: methanol = 1: 0 ～ 10: 1) to obtain the target intermediate. Body D114 (1.2g, yield: 40%), character: white solid.

LCMS: Rt: 1.442 min; MS m / z (ESI): 586.5 [M + H].

Intermediate Preparation Example 115: Preparation of Intermediate D115

At room temperature, Lawson's reagent (276 mg, 0.683 mmol) was added to a solution of D114 (400 mg, 0.683 mmol) in toluene (10 mL), and the reaction solution was stirred at 110 ° C for 4 hours. After the reaction is complete, the reaction solution is cooled to room temperature, filtered, and the filtrate is concentrated under reduced pressure, and the residue is purified by silica gel column chromatography (eluent gradient: dichloromethane: methanol = 1: 0 ～ 10: 1) to obtain the target Intermediate D115 (250 mg, yield: 49%), properties: white solid.

LCMS: Rt: 1.845 min; MS m / z (ESI): 602.4 [M + H].

Intermediate Preparation Example 116: Preparation of Intermediate D116

At room temperature, trifluoroacetic acid (3 mL) was added to a solution of D115 (250 mg, 0.41 mmol) in methylene chloride (12 mL), and the reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude target compound D116 (180 mg, yield: 86%), character: white solid, which was directly used in the next step.

LCMS: Rt: 1.006 min; MS m / z (ESI): 502.3 [M + H].

Intermediate Preparation Example 117: Preparation of Intermediate D117

At room temperature, D84 (2.0 g, 8.47 mmol) and palladium / carbon catalyst (300 mg) were added to ethanol (30 mL), the reaction liquid was replaced with hydrogen three times, and then stirred at 50 ° C. for 1 hour. The reaction was detected by LCMS, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the target compound D117 (1.6 g, crude product), character: yellow solid.

LCMS: Rt: 0.49 min; MS m / z (ESI): 237.2 [M + H].

Intermediate Preparation Example 118: Preparation of Intermediate D118

D117 (1.52 mg, 7.91 mmol), and D12 (2.5 g, 7.91 mmol) and ammonium chloride (2.09 g, 39.55 mmol) were dissolved in ethanol (30 mL). The reaction solution was stirred at 100 ° C for 8 hours. The reaction was detected to be complete by LCMS. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain the target compound D118 (4.0 g, crude product). Properties: yellow solid.

LCMS: Rt: 1.665 min; (0.1% FA-H ₂ O): (0.1% FA-ACN) = 90: 10; MS m / z (ESI): 572.5 [M + H].

Intermediate Preparation Example 119: Preparation of Intermediate D119

D118 (2.0 g, 3.5 mmol) was dissolved in a solution of hydrogen chloride in dioxane (20 mL) and methanol (5 mL), and the reaction mixture was stirred at room temperature for 2 hours. LCMS detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure to obtain the crude target compound D119 (2.1 g, crude product), character: yellow solid.

LCMS: Rt: 1.165 min; (0.1% FA-H ₂ O): (0.1% FA-ACN) = 90: 10; MS m / z (ESI): 472.2 [M + H].

Intermediate Preparation Example 120: Preparation of Intermediate D120

At room temperature, trifluoroacetic acid (2 mL) was added to a solution of D114 (100 mg, 0.17 mmol) in methylene chloride (8 mL), and the reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude target compound D120 (72 mg, yield: 86%), ZZ property: white solid, which was directly used in the next step.

LCMS: Rt: 0.748 min; MS m / z (ESI): 486.2 [M + H].

Intermediate Preparation Example 121: Preparation of Intermediate D121

D54 (200 mg, 0.54 mmol) was dissolved in trifluoroacetic acid (1 mL) and dichloromethane (5 mL), and the reaction mixture was reacted at room temperature for 2 hours. LCMS detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure to obtain the target compound D121 (140 mg, crude product), character: yellow solid.

LCMS: Rt: 0.94 min; (0.1% FA-H ₂ O): (0.1% FA-CAN) = 90: 10; MS m / z (ESI): 271.1 [M + H].

Intermediate Preparation Example 122: Preparation of Intermediate D122

Dissolve D121 (100 mg, 0.37 mmol), 1-bromo-2-methoxyethane (101 mg, 0.74 mmol) and N, N-diisopropylethylamine (143 mg, 1.11 mmol) in acetonitrile (5 mL), The reaction solution was stirred at 70 ° C for 10 hours. LCMS detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was purified by thin-layer preparative chromatography (eluent gradient: ethyl acetate: petroleum ether = 1: 1, R _f = 0.6) to obtain the target compound D122 (95mg, yield: 78.5%), character: white solid.

LCMS: Rt: 1.08 min; (0.1% FA-H ₂ O): (0.1% FA-CAN) = 90: 10; MS m / z (ESI): 329.1 [M + H].

Intermediate Preparation Example 123: Preparation of Intermediate D123

D122 (90 mg, 0.27 mmol), ammonium chloride (14 mg, 0.27 mmol) and iron powder (15 mg, 0.27 mmol) were added to water (2 mL) and ethanol (10 mL), and the reaction mixture was heated to 80 ° C. for 8 hours. LCMS detected the reaction was complete, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D123 (70 mg, crude), character: brown solid.

LCMS: Rt: 0.375 min; (0.1% FA-H ₂ O): (0.1% FA-CAN) = 90: 10; MS m / z (ESI): 299.4 [M + H].

Intermediate Preparation Example 124: Preparation of Intermediate D124

At room temperature, D71 (1.7 g, 6.38 mmol), D12 (2 g, 6.38 mmol) and ammonium chloride (3.4 g, 63.8 mmol) were dissolved in ethanol (30 mL), and the reaction was stirred at 80 ° C. for 12 hours. After the reaction was complete, the reaction mixture was poured into water (100 mL). The mixture was extracted with dichloromethane / methanol (dichloromethane / methanol = 10/1, 30 mL × 3), the organic phase was dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent gradient: dichloromethane: methanol = 20: 1, Rf = 0.5) to obtain the target product D124 (2g, yield: 57%), character: yellow solid.

LCMS: Rt: 1.562 min; MS m / z (ESI): 546.5 [M + H].

Intermediate Preparation Example 125: Preparation of Intermediate D125

At room temperature, D124 (1 g, 1.8 mmol) was dissolved in a mixed solution of hydrogen chloride in dioxane solution (4M, 10 mL) and methanol (10 mL). After the reaction was stirred at room temperature for 1 hour, the mixture was concentrated under reduced pressure to obtain Target compound D125 (800 mg, yield: 94%), properties: white solid.

LCMS: Rt: 0.796 min; MS m / z (ESI): 446.3 [M + H].

Intermediate Preparation Example 126: Preparation of Intermediate D126

At room temperature, 2-chloro-N, N-dimethylethylamine hydrochloride (3g, 21.23mmol), 4-nitro-1H-pyrazole (2g, 17.69mmol), cesium carbonate (17.29g, 53mmol) ) And diisopropylethylamine (4.56 g, 35.38 mmol) were mixed in N′N-dimethylformamide (30 mL), and the reaction was stirred at 130 ° C. for 12 hours. After the reaction was completed, the reaction mixture was poured into water (300 mL); the mixture was extracted with dichloromethane / methanol (10/1, 100 mL × 3), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent gradient: dichloromethane: methanol = 20: 1, Rf = 0.4) to obtain the target product D126 (2.2 g, yield: 68%), properties: yellow solid .

LCMS: Rt: 0.394 min; MS m / z (ESI): 185.3 [M + H].

Intermediate Preparation Example 127: Preparation of Intermediate D127

At room temperature, D126 (500 mg, 2.71 mmol) and palladium-carbon catalyst (150 mg) were added to methanol (10 mL), the reaction system was replaced with hydrogen three times, and then the reaction solution was stirred under hydrogen at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the target product D127 (400 mg, yield: 96%), character: yellow solid.

LCMS: Rt: 0.285 min; MS m / z (ESI): 155.2 [M + H].

Intermediate Preparation Example 128: Preparation of Intermediate D128

At room temperature, D21 (400 mg, 0.71 mmol) and Lawesson's reagent (578 mg, 1.42 mmol) were dissolved in toluene (10 mL), and the reaction solution was stirred at 120 ° C. for 3 hours. The product was detected by LCMS, the reaction solution was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent gradient: methanol / dichloromethane = 0-10%) to obtain the target product D128 (200 mg, Rate: 50%), traits: light yellow solid.

LCMS: Rt: 2.25 min; MS m / z (ESI): 576.2 [M + H].

Intermediate Preparation Example 129: Preparation of Intermediate D129

At room temperature, D128 (100 mg, 0.17 mmol) was dissolved in a 4M solution of hydrogen chloride in dioxane (0.4 mL, 1.7 mmol), and the reaction solution was stirred at room temperature for 12 hours. LCMS detected that the reaction was complete, and the reaction mixture was concentrated under reduced pressure to obtain the crude target product D129 (81 mg, yield: 98%), character: light yellow solid.

LCMS: Rt: 2.10 min; MS m / z (ESI): 476.2 [M + H].

Intermediate Preparation Example 130: Preparation of Intermediate D130

At room temperature, D118 (600 mg, 1.0 mmol) and Lawson's reagent (808 mg, 2.0 mmol) were dissolved in toluene (30 mL), and the reaction solution was heated to 110 ° C. and stirred for 4 hours. After the reaction was completed, the reaction mixture was poured into water (100 mL), the mixture was extracted with dichloromethane / methanol (10/1, 30 mL × 3), the organic phase was dried and filtered with anhydrous Na ₂ SO ₄ , the filtrate was concentrated under reduced pressure, and the residue Purified by silica gel column chromatography (eluent gradient: dichloromethane: methanol = 20: 1, Rf = 0.8) to obtain the target product D130 (300 mg, yield: 49%), character: white solid.

LCMS: Rt: 1.894 min; MS m / z (ESI): 588.5 [M + H].

Intermediate Preparation Example 131: Preparation of Intermediate D131

At room temperature, D130 (100 mg, 0.17 mmol) was dissolved in a mixture of hydrogen chloride in dioxane solution (4M, 2 mL) and methanol (1 mL). After the reaction was stirred at room temperature for 1 hour, the mixture was concentrated under reduced pressure to obtain Target compound D131 (80 mg, yield: 95%), character: yellow solid.

LCMS: Rt: 1.225 min; MS m / z (ESI): 488.1 [M + H].

Intermediate Preparation Example 132: Preparation of Intermediate D132

At room temperature, dissolve tert-butyl 9-oxo-3-azaspiro [5.5] undecane-3-carboxylate (500 mg, 1.87 mmol) in methanol (10 mL), then add sodium borohydride in portions (171 mg, 4.5 mmol), the reaction solution was stirred at room temperature for 1 hour. LCMS detected the completion of the reaction. To the system was added saturated aqueous sodium bicarbonate solution (50 mL), followed by extraction with ethyl acetate (20 mL * 3). The combined organic phase was washed with saturated brine (50 mL), dried and filtered with anhydrous sodium sulfate The filtrate was concentrated under reduced pressure to obtain crude target product D132 (500 mg, yield: 60%), character: colorless oil.

LCMS: Rt: 1.75 min; MS m / z (ESI): 214.5 [M-56 + H].

Intermediate Preparation Example 133: Preparation of Intermediate D133

Under ice bath, dissolve D132 (500 mg, 1.87 mmol) in dichloromethane (20 mL), add triethylamine (0.77 mL, 5.61 mmol), and then add methylsulfonyl chloride (0.29 mL, 3.74 mmol); The reaction was stirred at room temperature for 12 hours. The reaction was quenched with saturated aqueous sodium bicarbonate (50 mL), extracted with dichloromethane (20 mL × 3), the combined organic phases were washed with saturated brine (50 mL), then dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. Column chromatography (eluent gradient: petroleum ether / ethyl acetate = 5: 1) was purified to obtain the target product D133 (100 mg, yield: 16%), character: colorless oil.

LCMS: Rt: 1.89 min; MS m / z (ESI): 292.4 [M-56 + H].

Intermediate Preparation Example 134: Preparation of Intermediate D134

At room temperature, D133 (90 mg, 0.26 mmol), 4-nitro-1H-pyrazole (58 g, 0.52 mmol) and potassium carbonate (108 mg, 0.78 mmol) were added to N, N-dimethylformamide (2 mL) The reaction solution was stirred at 110 ° C for 12 hours. LCMS detected product formation, the reaction solution was diluted with water (20 mL), the compound was extracted with ethyl acetate (40 mL × 4), the organic phase was combined and washed with saturated brine (100 mL × 2), then dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative thin layer chromatography (eluent gradient: methanol / dichloromethane = 1:10) to obtain the target product D134 (50 mg, yield: 52%), character: white solid.

LCMS: Rt: 2.04 min; MS m / z (ESI): 309.3 [M-56 + H].

Intermediate Preparation Example 135: Preparation of Intermediate D135

At room temperature, combine tert-butyl 9- (4-nitro-1H-pyrazol-1-yl) -3-azaspiro [5.5] undecane-3-carboxylate (50 mg, 0.14 mmol) and palladium A carbon catalyst (30 mg) was added to absolute ethanol (3 mL), and the reaction liquid was replaced with hydrogen three times and stirred at room temperature for 2 hours. The reaction was detected by LCMS, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude target compound D135 (48 mg, yield: 98%), character: light yellow solid.

LCMS: Rt: 1.16 min; MS m / z (ESI): 335.2 [M + H].

Intermediate Preparation Example 136: Preparation of Intermediate D136

At room temperature, D135 (48 mg, 0.14 mmol), D12 (44 mg, 0.14 mmol) and ammonium chloride (38 mg, 0.70 mmol) were dissolved in isopropanol (3 mL), and the reaction solution was stirred at 80 ° C. for 3 hours. LCMS detected the reaction was complete, the reaction solution was diluted with saturated aqueous sodium bicarbonate solution (10 mL), then extracted with ethyl acetate (20 mL × 3), the combined organic phase was washed with saturated brine (50 mL), and then dried and filtered over anhydrous sodium sulfate The filtrate was concentrated under reduced pressure, and the residue was purified by preparative thin layer chromatography (eluent gradient: methanol / dichloromethane = 1:10) to obtain the target product D136 (32 mg, yield: 37%), properties: white solid.

LCMS: Rt: 1.89 min; MS m / z (ESI): 614.2 [M-56 + H].

Intermediate Preparation Example 137: Preparation of Intermediate D137

At room temperature, D136 (32 mg, 0.05 mmol) was dissolved in a 4 M solution of hydrogen chloride in dioxane (0.13 mL, 0.5 mmol), and the reaction solution was stirred at room temperature for 3 hours. LCMS detected that the reaction was complete, and the reaction mixture was concentrated under reduced pressure to obtain the crude target product D137 (30 mg, yield: 98%), character: light yellow solid.

LCMS: Rt: 0.37 min; MS m / z (ESI): 514.2 [M + H].

Intermediate Preparation Example 138: Preparation of Intermediate D138

At room temperature, 3- (benzyloxy) cyclobutanone (5g, 28.4mmol) and tert-butylpiperazine-1-carboxylate (5.28g, 28.4mmol) were dissolved in 1,2-dichloroethane (500 mL), sodium triacetoxyborohydride (11.98 g, 56.8 mmol) was added in portions, and the reaction solution was stirred at room temperature for 8 hours. The reaction was completed by LCMS. Water (100 mL) was added to the system. The mixture was extracted with dichloromethane (200 mL × 4). The combined organic phase was dried and filtered over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure. The crude product was subjected to silica gel column chromatography (Eluent gradient: petroleum ether / ethyl acetate = 100: 1 to 2: 1) Purification to obtain the target compound D138 (8.25 g, yield: 83.9%), character: white solid.

LCMS: Rt: 1.345 min; (0.1% FA-H ₂ O): (0.1% FA-ACN) = 90: 10; MS m / z (ESI): 347.5 [M + H].

Intermediate Preparation Example 139: Preparation of Intermediate D139

At room temperature, D138 (6.0 g, 17.34 mmol) was dissolved in trifluoroacetic acid (20 mL), and the reaction solution was stirred at 80 ° C. for 6 hours. LCMS detected the reaction was complete, the reaction mixture was concentrated to obtain a crude product, the crude product was dissolved in dichloromethane (5mL) and then added triethylamine (5.25g, 52.02mmol) and di-tert-butyl dicarbonate (7.56g, 34.68mmol) The reaction mixture was stirred at room temperature for 2 hours. The product was detected by LCMS, water (100 mL) was added to the system, the compound was extracted with dichloromethane (50 mL × 3), the organic phase was combined and concentrated under reduced pressure, and the resulting crude product was subjected to silica gel column chromatography (eluent gradient: two Chloromethane / methanol = 100: 1 ～ 10: 1) purification, to obtain the target compound D139 (3.9g, yield: 88.04%), properties: yellow oil.

LCMS: Rt: 1.006 min; (0.1% FA-H ₂ O): (0.1% FA-ACN) = 90: 10; MS m / z (ESI): 257.5 [M + H].

Intermediate Preparation Example 140: Preparation of Intermediate D140

Dissolve D139 (3.0g, 11.71mmol) in dichloromethane (50mL) and add triethylamine (3.5g, 35.13mmol), then add methylsulfonyl chloride (2.6mg, 23.43mmol); the reaction solution was stirred at room temperature 10 hours. Water (50 mL) was added to the system, the mixture was extracted with dichloromethane (50 mL × 4), the combined organic phases were washed with saturated aqueous sodium bicarbonate solution (50 mL × 3), then dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure To obtain the crude target compound D140 (4.0g, crude), traits: light yellow liquid.

LCMS: Rt: 1.145 min; (0.1% FA-H ₂ O): (0.1% FA-ACN) = 90: 10; MS m / z (ESI): 335.3 [M + H].

Intermediate Preparation Example 141: Preparation of Intermediate D141

At room temperature, D140 (1.5g, 4.49mmol) and 4-nitro-1H-pyrazole (507mg, 4.49mmol), potassium carbonate (1.85g, 4.49mmol) was added to N'N-dimethylformamide (20mL ), The reaction solution was stirred at 110 ° C for 10 hours. LCMS detected the reaction was complete, diluted with water (50 mL), the mixture was extracted with ethyl acetate (30 mL × 4), the combined organic phase was washed with saturated aqueous potassium carbonate solution (50 mL × 3), then dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure The obtained crude product was purified by silica gel column chromatography (eluent gradient: ethyl acetate / petroleum ether = 1: 5 ～ ethyl acetate / methanol = 10: 1) to obtain the target compound D141 (650 mg, yield: 43.33 %), Character: white solid.

¹ H NMR (400 MHz, MeOD): δ 8.63 (s, 1H), 8.16 (s, 1H), 4.94-4.86 (m, 1H), 3.46 (brs, 4H), 3.29-3.16 (m, 1H), 2.63-2.57 (m, 4H), 2.39-2.36 (m, 4H), 1.47 (s, 9H).

Intermediate Preparation Example 142: Preparation of Intermediate D142

At room temperature, D141 (650 mg, 1.834 mmol) and palladium / carbon catalyst (300 mg) were added to tetrahydrofuran (30 mL), the reaction liquid was replaced with hydrogen three times, and then stirred at room temperature for 10 hours. The reaction was completed by LCMS, and the palladium / carbon was filtered off. The filtrate was concentrated under reduced pressure to obtain the target compound D142 (610 mg, crude product). Properties: pale yellow solid.

LCMS: Rt: 0.303 min; (0.1% FA-H ₂ O): (0.1% FA-ACN) = 90: 10; MS m / z (ESI): 322.2 [M + H].

Intermediate Preparation Example 143: Preparation of Intermediate D143

D142 (509 mg, 1.58 mmol), D12 (500 g, 1.58 mmol) and ammonium chloride (334 g, 6.32 mmol) were dissolved in ethanol (5 mL); the reaction solution was stirred at 110 ° C. for 10 hours. LCMS detected that the reaction was complete, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent gradient: dichloromethane / methanol = 100: 1-10: 1) to obtain the target compound D143 (400mg, yield: 42.1%), character: white solid.

LCMS: Rt: 1.250 min; (0.1% FA-H2O): (0.1% FA-ACN) = 90:10; MS m / z (ESI): 601.2 [M + H].

Intermediate Preparation Example 144: Preparation of Intermediate D144

D143 (150 mg, 0.25 mmol) was dissolved in a solution of hydrogen chloride in dioxane (5 mL) and methanol (1 mL), and the reaction was stirred at room temperature for 1.5 hours. LCMS detected the reaction was complete, and the reaction solution was concentrated under reduced pressure to obtain the target compound D144 (130 mg, crude product), character: yellow solid.

LCMS: Rt: 0.780 min; (0.1% FA-H2O): (0.1% FA-ACN) = 90: 10; MS m / z (ESI): 501.2 [M + H].

Intermediate Preparation Example 145: Preparation of Intermediate D145

At room temperature, D143 (200 mg, 0.33 mmol) and Lawesson's reagent (271 mg, 0.67 mmol) were dissolved in toluene (10 mL), and the reaction was stirred at 110 ° C for 4 hours. After the reaction was completed, the reaction mixture was poured into water (100 mL), extracted with dichloromethane / methanol (10/1, 30 mL × 3), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure; the residue was subjected to silica gel column chromatography Chromatography (eluent gradient: dichloromethane: methanol = 20: 1, Rf = 0.6) was purified to obtain the target product D145 (120 mg, yield: 59%), character: yellow solid.

LCMS: Rt: 1.019 min; MS m / z (ESI): 517.1 [M-100 + H].

Intermediate Preparation Example 146: Preparation of Intermediate D146

At room temperature, D145 (120 mg, 0.19 mmol) was dissolved in a mixture of hydrogen chloride in dioxane solution (4M, 4 mL) and methanol (1 mL). After the reaction was stirred at room temperature for 1 hour, the mixture was concentrated under reduced pressure to obtain Crude target compound D146 (100 mg, yield 99%), character: yellow solid.

LCMS: Rt: 2.118 min; MS m / z (ESI): 517.2 [M + H].

Example 1 and Example 1 ’

Dimethyl (2-((2-((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) -5- (trifluoro Methyl) pyrimidin-4-yl) amino) phenyl) oxidation Phosphine and dimethyl (2-((4-((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) -5- ( Trifluoromethyl) pyrimidin-2-yl) amino) phenyl) Phosphine oxide

A mixture of compound D10 and D10 '(140 mg, 0.42 mmol) and compound D9 (79 mg, 0.44 mmol) were dissolved in EtOH (10 mL), and after adding ammonium chloride (106.9 mg, 2.0 mmol) to the reaction system, the reaction was Stir at 105 ° C for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with ethanol (20 mL), the filtrate was concentrated under reduced pressure, and the residue was used for preparative high performance liquid chromatography (eluent gradient:

Gradient conditions:

) Purification to obtain pure product E1 '(trifluoroacetate salt, 14.5 mg, yield: 7.3%) and product E1 (trifluoroacetate salt, 74.5 mg, yield: 37.6%). E1 ’:

HPLC: 99.152%@214nm, 99.42%@254nm

LCMS: MS / m (zI): 495.3 [M + H]

¹⁹ F NMR (400MHz, CD ₃ OD) δ: -62.90, -63.28, -77.23.

¹ H NMR (400 MHz, CD ₃ OD) δ: 8.38 (brs, 1H), 8.15 (brs, 1H), 7.87-7.68 (m, 2H), 7.52-7.35 (m, 2H), 4.43-4.27 (m, 1H), 4.08-4.04 (m, 2H), 2.58 (brs, 2H), 2.24-2.12 (m, 5H), 1.87-1.76 (m, 8H).

E1:

HPLC: 99.468%@214nm, 99.228%@254nm

LCMS: MS / m (zI): 495.3 [M + H]

¹⁹ F NMR (400MHz, CD ₃ OD) δ: -64.17, -77.37.

¹ H NMR (400 MHz, CD ₃ OD) δ: 8.35 (s, 1H), 7.94-7.90 (m, 1H), 7.60-7.54 (m, 1H), 7.49 (s, 1H), 7.41-7.37 (m, 1H) 7.28-7.24 (m, 1H), 4.40-4.33 (m, 1H), 4.10-4.06 (m, 2H), 3.62-3.56 (m, 2H), 2.27-2.16 (m, 2H), 2.14 (s , 3H), 1.84-1.81 (m, 8H).

Example 2

Dimethyl (2-((2-((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) -5- (trifluoro Methyl) pyrimidin-4-yl) amino) phenyl) oxidation phosphine

Compound D11 (100 mg, 0.6 mmol) and intermediate D12 (158 mg, 0.5 mmol) were dissolved in isopropanol (20 mL) at room temperature. After adding trifluoroacetic acid (1.14 g, 10.0 mmol) to the reaction system, the reaction The solution was heated to 120 ° C and stirred for 3 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was subjected to preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E2 (trifluoroacetate salt, 29.1 mg, yield: 13%, white solid).

HPLC: 99.83%@214nm, 99.81%@254nm

LCMS: MS / m (zI): 447.4 [M + H]

¹⁹ F NMR (376.5MHz, methanol-d ₄ ) δ-74.71 (s)

¹ H NMR (400MHz, DMSO-d ₆ ) δ 10.94 (brs, 1H), 9.42 (brs, 1H), 8.17 (brs, 1H), 7.65-7.45 (m, 4H), 7.25 (brs, 1H), 4.26 (brs, 1H), 3.97-3.93 (m, 2H), 3.45 (t, J = 11.2 Hz, 2H), 1.89-1.75 (m, 10H).

Example 3

(2-((5-chloro-2-((5-chloro-1- (2-hydroxyethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) di Methylphosphine oxide (E3)

Compound D58 (165 mg, 0.6 mmol) and intermediate D12 (158 mg, 0.5 mmol) were dissolved in isopropanol (20 mL) at room temperature. After adding trifluoroacetic acid (1.14 g, 10.0 mmol) to the reaction system, the reaction The solution was heated to 120 ° C and stirred for 3 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was subjected to preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E3 (trifluoroacetate salt, 15.3 mg, yield: 16%), character: white solid.

HPLC: 99.11%@214nm, 98.96%@254nm

LCMS: MS / m / z (ESI): 441.3 [M + H]

¹ H NMR (400MHz, methanol-d ₄ ) δ8.4 (br, 1H), 8.07 (s, 1H), 7.68-7.58 (m, 3H), 7.37-7.33 (m, 1H), 4.27 (t, J = 5.6 Hz, 2H), 3.92 (t, J = 5.6 Hz, 2H), 1.89 (s, 3H), 1.85 (s, 3H).

¹⁹ F NMR (376.5MHz, methanol-d ₄ ) δ-77.25 (s)

Example 4

(5-chloro-2 -– (2- (1- (2-hydroxyethyl) -1-hydropyrazole-4-amino) pyrimidine 4-amino) phenyl) dimethyl phosphine oxide

Compound D17 (100 mg, 0.70 mmol) and intermediate D12 (223.0 mg, 0.7 mmol) were dissolved in isopropanol (5 mL) at room temperature. After adding trifluoroacetic acid (1596.0 mg, 14.0 mmol) to the reaction system, the reaction The reaction was stirred at 120 ° C for 3 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with ethanol (20 mL), the filtrate was concentrated under reduced pressure, and the residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E4 (trifluoroacetate salt, 47.4 mg, yield: 16%, white solid).

HPLC: 99.84%@214nm, 99.83%@254nm

LCMS: MS / m (zI): 421.2 [M + H]

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.98 (brs, 1H), 9.38 (s, 1H), 9.05–7.84 (m, 1H), 7.96–7.30 (m, 4H), 7.22 (brs, 1H ), 4.03 (brs, 2H), 3.69 (t, J = 5.3 Hz, 2H), 1.76 (s, 3H), 1.79 (s, 3H),

Example 5

(2- (5-chloro-2-((1- (2-methoxyethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine Oxide

After the compound D18 (100 mg, 0.4 mmol) and the intermediate D12 were completed at room temperature, the reaction solution was stirred at 80 ° C for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with ethanol (20 mL), the filtrate was concentrated under reduced pressure, and the residue was used for preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E5 (trifluoroacetate salt, 27.4 mg, yield: 9.7%) Properties: white solid.

HPLC: 90.63%@214nm, 96.38%@254nm

LCMS: MS / m (zI): 407.1 [M + H]

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.11 (br, 1H), 9.42 (s, 1H), 8.17 (brs, 2H), 7.89-7.52 (m, 3H), 7.45 (s, 1H), 7.24 (brs, 1H), 4.15 (brs, 2H), 3.63 (t, J = 5.2 Hz, 2H), 3.20 (s, 3H), 1.79 (s, 3H), 1.76 (s, 3H).

Example 6

(2-((5-chloro-2-((5-chloro-1- (2-methoxyethyl) -1H-pyrazol-4-yl) pyrazol-4-yl) pyrimidin-4-yl ) Pyrazol-4-yl) phenyl) dimethylphosphine oxide

Compound D20 (100 mg, 0.57 mmol) and intermediate D12 (150 mg, 0.475 mmol) were dissolved in isopropanol (20 mL) at room temperature, and trifluoroacetic acid (1.1 g, 9.5 mmol) was added to the reaction system. The reaction was heated to 120 ° C and stirred for 3 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E6 (hydrochloride, 29.4 mg, yield: 13.6%), character: white solid.

HPLC: 98.83%@214nm, 99.17%@254nm

LCMS: MS / m (zI): 455.4 [M + H]

¹ H NMR (400MHz, MeOH-d ₄ ) δ8.5 ～ 8.11 (m, 2H), 7.73-7.60 (m, 3H), 7.44-7.41 (m, 1H), 4.35 (brs, 2H), 3.79 (brs , 2H), 3.30 (s, 3H), 1.90 (s, 3H), 1.87 (s, 3H)

Example 7

(2-((5-methyl-2-((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) benzene Radical) dimethyl phosphine oxide

Compound D21 (180 mg, 0.32 mmol) was dissolved in a mixed solvent of HCl-dioxane (3 mL) and methanol (1 mL) at room temperature, and the reaction was stirred at room temperature for 1 hour. After LCMS detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was subjected to preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E7 (HCl salt, 16 mg, yield: 32%), character: white solid;

HPLC: 99.084%@214nm, 99.750%@254nm

LCMS: MS / m (zI): 460.4 [M + H]

¹ H NMR (400 MHz, MeOD-d ₄ ) δ: 8.56 (brs, 1H), 8.12-7.98 (m, 1H), 7.68 (brs, 2H), 7.55 (s, 1H), 7.40 (brs, 1H), 4.60 (brs, 1H), 3.58 (d, J = 12.8 Hz, 2H), 3.26-3.30 (m, 2H), 2.22-2.34 (m, 7H), 1.91 (s, 3H), 1.88 (s, 3H) .

Example 8

(2-((5-chloro-2-((5-chloro-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) Amino) phenyl) dimethylphosphine oxide

At room temperature, compound D23 (100 mg, 0.5 mmol) and intermediate D12 (131 mg, 0.42 mmol) were dissolved in isopropanol (20 mL), trifluoroacetic acid (960 mg, 8.4 mmol) was added to the reaction system, and the reaction solution was heated The reaction was stirred at 120 ° C for 3 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was subjected to preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E8 (hydrochloride, 35.9 mg, yield: 17.8%) Properties: white solid.

HPLC: 98.84%@214nm, 99.86%@254nm

LCMS: MS / m (zI): 481.3 [M + H]

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 8.6-8.1 (m, 2H), 7.74-7.67 (m, 3H), 7.44-7.41 (m, 1H), 4.61 (brs, 1H),

4.10-4.06 (m, 2H), 3.63-3.56 (m, 2H), 2.23-2.16 (m, 2H), 1.91-1.87 (m, 8H).

Example 9

(((5-chloro-2– (2- (1- (2-hydroxyethyl) -5-methyl-1-hydropyrazole-4-amino) pyrimidine 4-amino) phenyl) amino) phenyl ) Dimethylphosphine oxide (E9)

At room temperature, compound D25 (100 mg, 0.39 mmol) and intermediate D12 (148 mg, 0.47 mmol) were dissolved in isopropanol (10 mL), and trifluoroacetic acid (889.2 mg, 7.8 mmol) was added to the reaction system. The reaction was stirred at ℃ for 3 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with ethanol (20 mL), the filtrate was concentrated under reduced pressure, and the residue was used for preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E9 (trifluoroacetate salt, 22.4 mg, yield: 11%), character: white solid.

HPLC: 99.20%@214nm, 99.82%@254nm

LCMS: MS / m (zI): 421.2 [M + H]

¹ H NMR (400 MHz, MeOD) δ 8.47 (brs, 1H), 7.95 (s, 1H), 7.64–7.42 (m, 3H), 7.19-7.22 (m, 1H), 4.15 (t, J = 5.4Hz , 2H), 3.87 (t, J = 5.4Hz, 2H), 2.18 (s, 3H), 1.86 (s, 3H), 1.83 (s, 3H)

Example 10

(2-((5-chloro-2-((1- (2-methoxyethyl) -5-methyl-1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) benzene Radical) dimethyl phosphine oxide

Compound D27 (155 mg, 1.0 mmol) and intermediate D12 (158 mg, 0.5 mmol) were dissolved in isopropanol (20 mL) at room temperature, and trifluoroacetic acid (1.14 g, 10.0 mmol) was added to the reaction system. The reaction was heated to 120 ° C and stirred for 3 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E10 (trifluoroacetate salt, 35.1 mg, yield: 16.2%), character: white solid.

HPLC: 96.49%@214nm, 98.48%@254nm

LCMS: MS / m (zI): 435.4 [M + H]

¹⁹ F NMR (376.5MHz, methanol-d4): δ-77.21 (s)

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 8.6-8.0 (m, 2H), 7.67-7.52 (m, 3H), 7.38 (brs, 1H), 4.27-4.25 (m, 2H), 3.75 (brs , 2H), 3.30 (s, 3H), 2.21 (s, 3H), 1.90 (s, 3H), 1.86 (s, 3H).

Example 11

(2- (5-chloro-2- (1- (1- (2-hydroxyethyl) piperidin-4-yl) -5-methyl-1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) aminophenyl) dimethylphosphine oxide

Compound D28 (30 mg, 0.056 mmol) was dissolved in tetrahydrofuran / methanol (1 mL / 1 mL) at room temperature, and sodium borohydride (11 mg, 0.28 mmol) was added to the reaction solution under ice bath conditions. The reaction solution was stirred at room temperature for two hours. After LCMS detected that the reaction was complete, the reaction mixture was poured into saturated aqueous ammonium chloride solution (5 mL), the mixture was extracted with ethyl acetate (10 mL × 4), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E11 (trifluoroacetate salt, 20 mg, yield: 50%). Properties: colorless oil.

HPLC: 93.04% (214nm), 93.66% (254nm)

LCMS: Rt: 0.70 min; MS m / z (ESI): 504.5 [M-H].

¹ H NMR (400MHz, MeOD) δ 8.43 (brs, 1H), 8.02 (brs, 1H), 7.64 (dd, J = 12.0Hz, 8.0Hz, 1H), 7.55 (brs, 2H), 7.33 (brs, 1H), 4.67-4.54 (m, 1H), 3.93-3.91 (m, 2H), 3.83-3.80 (m, 2H), 3.65-3.40 (m, 1H), 3.26-3.24 (m, 3H), 2.49- 2.45 (m, 2H), 2.24-2.17 (m, 5H), 1.89 (s, 3H), 1.85 (s, 3H).

Example 12

(2-((5-chloro-2-((5-methyl-1- (4-hydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Radical) amino) phenyl) dimethyl phosphine oxide

Compound D12 (95 mg, 0.30 mmol), compound D29 (50 mg, 0.27 mmol) and NH ₄ Cl (72 mg, 1.35 mmol) were dissolved in EtOH (3 mL) at room temperature, and the reaction solution was stirred at 100 ° C. for 10 hours. After the reaction was checked by LCMS, the reaction solution was cooled to room temperature, the mixture was filtered to remove NH ₄ Cl, and the filtrate was concentrated under reduced pressure. The residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E12 (HCl salt, 15 mg, yield: 12%). Properties: white solid;

HPLC: 99.574%@214nm, 99.705%@254nm

LCMS: MS / m (zI): 461.4 [M + H]

¹ H NMR (400 MHz, MeOD-d ₄ ) δ: 8.59 (brs, 1H), 8.12-7.97 (m, 1H), 7.70-7.68 (m, 2H), 7.54 (s, 1H), 7.42 (brs, 1H ), 4.45 (brs, 1H), 4.05-4.09 (m, 2H), 3.56-3.62 (m, 2H), 2.20-2.26 (m, 5H), 1.88-1.92 (m, 8H).

Example 13

(2-((5-chloro-2-((5-cyclopropyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Radical) amino) phenyl) dimethyl phosphine oxide

Compound D31 (80 mg, 0.386 mmol) and intermediate D12 (101 mg, 0.322 mmol) were dissolved in isopropanol (20 mL) at room temperature, trifluoroacetic acid (734 mg, 6.44 mmol) was added to the reaction system, and then the reaction solution The reaction was heated to 120 ° C and stirred for 3 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E13 (hydrochloride, 3.1 mg, yield: 2%), character: white solid.

HPLC: 97.42%@214nm, 97.22%@254nm

LCMS: MS / m (zI): 487.4 [M + H]

¹ H NMR (400MHz, MeOH-d ₄ ) δ 8.56-7.97 (m, 2H), 7.70-7.43 (m, 4H), 4.86-4.84 (m, 1H), 4.12-4.07 (m, 2H), 3.61 (t, J = 12.0Hz, 2H), 2.30-2.19 (m, 2H), 1.93-1.89 (m, 8H), 1.85-1.79 (m, 1H), 1.02-1.00 (m, 2H), 0.73-0.71 (m, 2H).

Example 14

(2-((5-chloro-2-((5-ethyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl ) Amino) phenyl) dimethylphosphine oxide

At room temperature, compound D32 (36 mg, 0.12 mmol) and intermediate D29 (43 mg, 0.24 mmol) were dissolved in isopropanol (10 mL), and trifluoroacetic acid (274 mg, 2.4 mmol) was added to the reaction system; after the addition The reaction solution was heated to 120 ° C and stirred for 3 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E14 (hydrochloride, 24.7 mg, yield: 46.3%) Properties: white solid.

HPLC: 97.09%@214nm, 96.73%@254nm

LCMS: MS / m (zI): 477.4 [M + H]

¹⁹ F NMR ^(376.5MHz, methanol -d ₄₎ δ161.97 (s)

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 8.70-7.91 (m, 2H), 7.71-7.57 (m, 3H), 7.42-7.38 (m, 1H), 4.51-4.43 (m, 1H), 4.11 --4.06 (m, 2H), 3.61 (t, J = 11.6 Hz, 2H), 2.27 (s, 3H), 2.27-2.18 (m, 2H), 1.92-1.86 (m, 8H).

Example 15

(2-((5-Bromo-2-((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl ) Amino) phenyl) dimethylphosphine oxide

Dissolve compound D33 (100 mg, 0.28 mmol) and compound D29 (51 mg, 0.28 mmol) in EtOH (10 mL) at room temperature in a 100 mL single-necked bottle, add trifluoroacetic acid (5 mL), and then heat the reaction solution to 80 The reaction was stirred at ℃ overnight. LCMS detected the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was subjected to preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the target compound E15 (trifluoroacetate salt, 8 mg, yield: 10%), character: white solid.

HPLC: 99.487%@214nm, 99.739%@254nm

LCMS: MS / m (zI): 507 [M + H]

¹⁹ F NMR (376.5MHz, DMSO) δ 74.35 (s)

¹ H NMR (400MHz, DMSO) δ 11.11 (brs, 1H), 8.73 (brs, 1H), 8.16 (s, 1H), 7.53-7.57 (m, 1H), 7.45 (s, 1H), 7.15 (brs , 1H), 7.01 (brs, 1H), 4.37-4.31 (m, 1H), 3.97-3.94 (m, 2H), 3.49-3.46 (m, 2H), 2.15 (s, 3H), 2.09-1.99 (m , 2H), 1.79-1.75 (m, 2H), 1.79 (s, 3H), 1.75 (s, 3H).

Example 16

(2-((5-Bromo-2-((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl ) Amino) phenyl) dimethylphosphine oxide

Compound D34 (80 mg, 0.25 mmol), compound D29 (45 mg, 0.25 mmol) and ammonium chloride (110 mg, 2.5 mmol) were dissolved in ethanol (5 mL) at room temperature. The reaction solution was stirred at 100 ° C for 12 hours. After LCMS detected that the reaction was complete, the reaction mixture was concentrated under reduced pressure. The residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E16 (hydrochloride, 6mg, yield: 5%), character: yellow solid

HPLC: 97.65% (214nm), 99.39% (254nm)

LCMS: Rt: 1.04 min; MS m / z (ESI): 457.4 [M + H].

¹ H NMR (400 MHz, MeOD) δ 8.66 (brs, 1H), 7.72–7.64 (m, 1H), 7.57 (s, 2H), 7.36 (t, J = 7.2 Hz, 2H), 4.47 (t, J = 11.6Hz, 1H), 4.09 (dd, J = 11.4, 4.0Hz, 2H), 3.94 (s, 3H), 3.61 (t, J = 11.7Hz, 2H), 2.31–2.17 (m, 5H), 1.89 (m, 8H).

Example 17

Dimethyl (2-((5-methyl-2-((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine -4-yl) amino) phenyl) phosphine oxide

Compound D35 (50 mg, 0.169 mmol), compound D29 (30 mg, 0.169 mmol) and NH ₄ Cl (27 mg, 0.507 mmol) were dissolved in EtOH (3 mL) at room temperature, and the reaction solution was heated to 100 ° C. and stirred for 8 hours. The reaction was completed by LCMS, the mixture was filtered, and the filtrate was concentrated under reduced pressure; the residue was subjected to preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E17 (HCl salt, 15 mg, yield: 20%). Properties: oily liquid;

HPLC: 98.434%@214nm, 98.466%@254nm

LCMS: MS / m (zI): 441.4 [M + H]

¹ H NMR (400 MHz, MeOD-d ₄ ) δ: 8.70 (br, 1H), 7.66-7.52 (m, 4H), 7.35 (br, 1H), 4.43 (br, 1H), 4.08 (dd, J = 11.2 Hz, 4.4Hz, 2H), 3.63-3.56 (m, 2H), 2.29-2.18 (m, 8H), 1.94-1.84 (m, 8H).

Example 18

2- (4- (4- (5-chloro-4-((2- (dimethylphosphoryl) phenyl) amino) pyrimidin-2-yl) amino) -5-methyl-1H-pyrazole- 1-yl) piperidin-1-ylacetamide

Compound D37 (104 mg, 0.2 mmol), ammonium chloride (22 mg, 0.4 mmol), HATU (114 mg, 0.3 mmol) were dissolved in DMF (2 mL) at room temperature. The reaction solution was stirred at room temperature for 12 hours. After LCMS detected that the reaction was complete, the reaction mixture was poured into water (5 mL). The mixture was extracted with ethyl acetate (10 mL × 4), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E18 (hydrochloride, 20mg, yield: 20%), properties: light yellow solid

HPLC: 99.79% (214nm), 99.89% (254nm)

LCMS: Rt: 0.66 min; MS m / z (ESI): 517.5 [M-H].

¹ H NMR (400MHz, MeOD) δ 8.57 (brs, 1H), 7.99 (brs, 1H), 7.71 (brs, 2H), 7.56 (brs, 1H), 7.43 (brs, 1H), 4.65-4.55 (m , 1H), 4.05 (s, 2H), 3.81-3.78 (m, 2H), 3.44-3.38 (m, 2H), 2.53-2.50 (m, 2H), 2.35-2.25 (m, 5H), 1.92-1.89 (m, 6H).

Example 19 and Example 20

Dimethyl (2-((2-((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) -5- (trifluoromethyl) pyrimidine- 4-yl) amino) phenyl) phosphine oxide and dimethyl Yl (2-((4-((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) -5- (trifluoromethyl) pyrimidine-2- Group) amino) phenyl) phosphine oxide

Compound D40 (450 mg, 0.76 mmol) was dissolved in 4M HCl / dioxane (20 mL) at room temperature, and the reaction solution was stirred at room temperature for 2 h. The reaction was detected by LCMS. The reaction solution was concentrated under reduced pressure to obtain the target crude product (400 mg), and 50 mg of the crude product was used for preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain pure target product E19 (hydrochloride, 12.3mg, yield: 24.6%) Properties: white solid.

HPLC: 99.58%@214nm, 99.85%@254nm, Rt: 6.543min

LCMS: MS / m (zI): 494.4 [M + H], Rt: 1.094min

¹ H NMR (400 MHz, methanol-d ₄ ) δ 8.44 (br, 1H), 7.79-7.77 (m, 1H), 7.69-7.64 (m, 1H), 7.53-7.46 (m, 2H), 7.43-7.41 (m, 1H), 4.57 (br, 1H), 3.61-3.57 (m, 2H), 3.32-3.24 (m, 2H), 2.36-2.32 (m, 2H), 2.16 (s, 3H), 2.13-2.09 (m, 2H), 1.86 (s, 3H), 1.83 (s, 3H).

¹⁹ F NMR (376.5 MHz, methanol-d ₄ ) δ 64.68 (s).

At room temperature, compound D41 (420 mg, 0.76 mmol) was dissolved in 4M HCl / dioxane (20 mL), and the reaction solution was stirred at room temperature for 2 h, and the reaction was completed by LCMS. The reaction solution was concentrated under reduced pressure to obtain the target crude product (400 mg), and 50 mg of the crude product was used for preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain pure target product E20 (hydrochloride, 14.6 mg, yield: 29.2%,) Properties: white solid.

HPLC: 99.79%@214nm, 99.93%@254nm, Rt: 5.625min

LCMS: MS / m (zI): 494.5 [M + H], Rt: 0.915min

¹⁹ F NMR (376.5 MHz, methanol-d ₄ ) δ 63.652 (s).

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 8.44 (brs, 1H), 7.79-7.77 (m, 1H), 7.68-7.64 (m, 1H), 7.53-7.46 (m, 2H), 7.43-7.41 (m, 1H), 4.57 (brs, 1H), 3.61-3.58 (m, 2H), 3.32-3.24 (m, 2H), 2.35-2.32 (m, 2H), 2.16 (s, 3H), 2.12-2.09 (m, 2H), 1.86 (s, 3H), 1.83 (s, 3H).

Example 21

(2- (5-chloro-2-((1- (1- (2-fluoroethyl) piperidin-4-yl) -5-methyl-1H-pyrazol-4-yl) amino) pyrimidine- 4-yl) aminophenyl) dimethylphosphine oxide

Compound E7 (180 mg, 0.36 mmol), 1-fluoro-2 iodoethane (75 mg, 0.43 mmol), DIEA (140 mg, 1.08 mmol) were dissolved in DMF (3 mL) at room temperature. The reaction solution was stirred at room temperature for 12 hours. After LCMS detected that the reaction had been completed, the reaction solution was directly used for preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E21 (30 mg, yield: 17%). Properties: white solid

HPLC: 97.37% (214nm), 97.47% (254nm)

LCMS: Rt: 0.76 min; MS m / z (ESI): 506.4 [M + H].

¹ H NMR (400MHz, DMSO) δ 11.18 (s, 1H), 8.48 (s, 2H), 8.05 (s, 1H), 7.60-7.49 (m, 1H), 7.43 (s, 1H), 7.31 (brs , 1H), 7.11 (brs, 1H), 4.61 (t, J = 4.9Hz, 1H), 4.50 (t, J = 4.9Hz, 1H), 4.13-3.98 (m, 1H), 3.00 (d, J = 11.4Hz, 2H), 2.70 (t, J = 4.9Hz, 1H), 2.63 (t, J = 4.9Hz, 1H), 2.20-2.17 (m, 2H), 2.13 (s, 3H), 2.06-2.04 ( m, 2H), 1.79-1.76 (m, 8H).

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.11 (br, 1H), 9.42 (s, 1H), 8.17 (brs, 2H), 7.89-7.52 (m, 3H), 7.45 (s, 1H), 7.24 (brs, 1H), 4.15 (brs, 2H), 3.63 (t, J = 5.2 Hz, 2H), 3.20 (s, 3H), 1.79 (s, 3H), 1.76 (s, 3H).

Example 22

(2-((5-chloro-2-((5-ethyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl ) Amino) phenyl) dimethylphosphine oxide

Compound D43 (150 mg, 0.73 mmol) and intermediate D12 (158 mg, 0.5 mmol) were dissolved in isopropanol (30 mL) at room temperature, and trifluoroacetic acid (1.14 g, 10.0 mmol) was added to the reaction system; the addition was complete After that, the reaction liquid was heated to 120 ° C and the reaction was stirred for 3 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E22 (hydrochloride, 14.6 mg, yield: 6.2%) Properties: yellow solid.

HPLC: 95.92%@214nm, 97.59%@254nm

LCMS: MS / m (zI): 475.4 [M + H]

¹ H NMR (400 MHz, methanol-d ₄ ) δ: 8.45-7.97 (m, 2H), 7.71-7.43 (m, 4H), 4.49-4.46 (m, 1H), 4.09–4.06 (m, 2H), 3.62 (t, J = 12.0 Hz, 2H), 2.78-2.72 (m, 2H), 2.32-2.21 (m, 2H), 1.92-1.88 (m, 8H), 1.17-1.15 (m, 3H).

¹⁹ F NMR (376.5MHz, methanol-d ₄ ) δ: N / A

Example 23 and Example 24

(2-((5-chloro-2-((1-((1s, 4s) 4- (hydroxy) cyclohexyl) -5-methyl-1H-pyrazol-4-yl) amino) pyridine-4 -Yl) amino) phenyl) dimethylphosphine oxide Thing (cis and trans)

A solution of compound D49 (200 mg, 0.42 mmol) in methanol (5 mL) was cooled with an ice bath, and then sodium borohydride (160 mg, 0.42 mmol) was slowly added to the reaction solution; the reaction was carried out at room temperature for 2 hours after the addition. After the reaction was completed, a saturated aqueous ammonium chloride solution (10 mL) was added to the reaction system to quench the reaction, the mixture was concentrated under reduced pressure, and the residue was extracted with dichloromethane / methanol (20 / 1,10 mL × 3). The organic phases were combined and concentrated directly under reduced pressure, and the residue was subjected to preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E23 (40 mg, 20% yield). Properties: white solid.

HPLC: 99.51%@214nm, 99.26%@254nm

LCMS: MS / m (zI): 475.4 [M + H]

¹ H NMR (400MHz, DMSO-d6) δ: 11.18 (s, 1H), 8.46 (brs, 2H), 8.05 (s, 1H), 7.57-7.52 (m, 1H), 7.40 (s, 1H), 7.20 (br, 1H), 7.13-7.10 (m, 1H), 4.65 (d, J = 4.4Hz, 1H), 4.06-4.03 (m, 1H), 3.50-3.48 (m, 1H), 2.12 (s, 3H ), 1.93-1.75 (m, 6H), 1.79 (s, 3H), 1.75 (s, 3H), 1.38-1.34 (m, 2H).

The purity of the other isomer is not good, and then use preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E24 (trifluoroacetate salt, 10 mg, yield: 5%) Properties: white solid.

HPLC: 96.45%@214nm, 96.83%@254nm

LCMS: MS / m (zI): 475.4 [M + H]

¹ H NMR (400 MHz, MeOH-d6) δ: 8.57 (br, 1H), 8.20 (br, 1H), 7.95-9.68 (m, 2H), 7.52-7.35 (m, 2H), 4.20-4.05 (m, 1H), 4.03 (br, 1H), 2.42-2.32 (m, 2H), 2.23-2.18 (m, 3H), 1.98-1.87 (m, 8H), 1.76-1.68 (m, 4H).

¹⁹ F NMR (376.5MHz, MeOH-d6) δ: -77.33 (s)

Example 25 and Example 26

(2-((2-((1- (1- (2-hydroxyethyl) piperidin-4-yl) -5-methyl-1H-pyrazol-4-yl) amino) -5- (tri Fluoromethyl) pyrimidin-4-yl) amino) yl) dimethylphosphine Compound and (2-((4-((1- (1- (2- (hydroxyethyl) piperidin-4-yl) -5-methyl-1H-pyrazol-4-yl) amino) -5- (Trifluoromethyl) pyrimidin-2-yl) amino) phenyl) di Methylphosphine oxide

Compound E19 (hydrochloride, 100 mg, 0.2 mmol) and bromoethanol (38 mg, 0.3 mmol) were dissolved in MeCN (30 mL) at room temperature, and K ₂ CO ₃ (110 mg, 0.8 mmol) was added to the reaction system. The solution was heated to 80 ° C and stirred for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with acetonitrile (10 mL X 2), the filtrate was concentrated under reduced pressure, and the residue was subjected to preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E25 (16.9 mg, yield: 15.5%) Properties: white solid.

HPLC: 95.79%@214nm, 95.88%@254nm, Rt: 6.415min.

LCMS: MS / m (zI): 538.5 [M + H], Rt: 1.096min

¹ H NMR (400 MHz, methanol-d ₄ ) δ 8.23 (s, 1H), 8.14-8.10 (m, 1H), 7.51-7.46 (m, 2H), 7.29 (t, J = 8.0 Hz, 1H), 7.11-7.08 (m, 1H), 4.14 (br, 1H), 3.71 (t, J = 6.0 Hz, 2H), 3.14-3.11 (m, 2H), 2.59 (t, J = 6.0 Hz, 2H), 2.30 -2.24 (m, 4H), 2.13 (s, 3H), 1.91-1.88 (m, 2H), 1.82 (s, 3H), 1.79 (s, 3H).

¹⁹ F NMR (376.5MHz, methanol-d ₄ ) δ-63.388 (s).

Compound E20 (100 mg, 0.2 mmol) and bromoethanol (38 mg, 0.3 mmol) were dissolved in acetonitrile (30 mL) at room temperature, K ₂ CO ₃ (110 mg, 0.8 mmol) was added to the reaction system, and the reaction solution was heated to 80 The reaction was stirred at ℃ for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with acetonitrile (10 mL X 2), the filtrate was concentrated under reduced pressure, and the residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E26 (23.4 mg, yield: 21.4%) Properties: white solid.

HPLC: 98.56%@214nm, 98.34%@254nm, Rt: 5.485min.

LCMS: MS / m (zI): 538.6 [M + H], Rt: 0.916min.

¹⁹ F NMR (376.5MHz, methanol-d ₄ ) δ-61.933 (s).

¹ H NMR (400MHz, MeOH-d ₄ ) δ 8.23 (s, 1H), 8.14-8.10 (m, 1H), 7.51-7.46 (m, 2H), 7.32-7.27 (m, 1H), 7.11-7.08 (m, 1H), 4.14 (br, 1H), 3.71 (t, J = 6.0 Hz, 2H), 3.14-3.11 (m, 2H), 2.59 (t, J = 6.0 Hz, 2H), 2.30-2.24 ( m, 4H), 2.13 (s, 3H), 1.91-1.88 (m, 2H), 1.82 (s, 3H), 1.79 (s, 3H).

Example 27

(2- (5-chloro-2- (5-methoxy-1- (4-hydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino] pyrimidin-4-yl ) Aminophenyl) dimethylphosphine oxide

At room temperature, compound D59 (50 mg, 0.25 mmol) and intermediate D12 (79.0 mg, 0.25 mmol) were dissolved in EtOH (5 mL). After adding NH ₄ Cl (81 mg, 1.5 mmol) to the reaction system, the reaction solution was heated up The reaction was stirred at 80 ° C for 16 hours. After the reaction, the reaction solution was filtered, the filter cake was washed with ethanol (10 mL), the filtrate was concentrated under reduced pressure, and the residue was prepared by preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E27 (trifluoroacetate salt, 15.3 mg, yield: 16%) Properties: white solid.

HPLC: 95.66%@214nm, 97.91%@254nm

LCMS: MS / m (zI): 477.4 [M + H]

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 7.96-8.45 (m, 2H), 7.58-7.81 (m, 2H), 7.33 (brs, 2H), 4.34-4.48 (m, 1H), 3.94–3.99 (m, 5H), 3.48 (t, J = 12.0 Hz, 2H), 2.03-2.12 (m, 2H), 1.83 (s, 3H), 1.79 (s, 3H), 1.76-1.79 (m, 2H)

Example 28 and Example 29

(2-((5-chloro-2-((1-((1s, 4s) 4- (dimethylamino) cyclohexyl) -5-methyl-1H-pyrazol-4-yl) amino) pyridine -4-yl) amino) phenyl) dimethylphosphine oxide Compound (cis and trans)

At room temperature, dimethylamine hydrochloride (257 mg, 3.2 mmol) and triethylamine (646 mg, 6.4 mmol) were added to anhydrous dichloromethane (20 mL) and stirred at room temperature for half an hour before adding compound D49 (150 mg, 0.32 mmol) After stirring at room temperature for half an hour, sodium cyanoborohydride (340 mg, 1.6 mmol) was added, and the reaction was stirred at room temperature overnight. After the reaction, the reaction solution was washed with a saturated aqueous ammonium chloride solution (10 mL × 2), and the organic phase was concentrated under reduced pressure. The residue was prepared by high performance liquid chromatography (eluent gradient:

Purification gave the target product E28 (trifluoroacetate salt, 40 mg, yield: 25%, white solid) and product E29 (trifluoroacetate salt, 40 mg, yield: 25%, white solid).

E28:

HPLC: 99.24%@214nm, 99.00%@254nm, Rt: 3.983min

LCMS: MS / m (zI): 501 [M + H], Rt: 1.104min

19F NMR (376.5MHz, methanol-d4) δ-76.98 (s)

¹ H NMR (400MHz, MeOH-d6) δ: 8.60-7.90 (m, 2H), 7.68-7.66 (m, 2H), 7.52 (s, 1H), 7.38 (br, 1H), 4.24 (br, 1H) , 3.39-3.30 (m, 1H), 2.90 (s, 6H), 2.22-2.10 (m, 5H), 2.09-2.05 (m, 4H), 1.90-1.78 (m, 8H).

E29:

HPLC: 98.76%@214nm, 98.62%@254nm, Rt: 4.165min

LCMS: MS / m (zI): 501 [M + H], Rt: 1.124min

19F NMR (376.5MHz, DMSO-d6) δ-74.70 (s)

¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 11.84 (br, 1H), 9.80-9.50 (m, 2H), 8.80-8.30 (m, 2H), 7.68-7.62 (m, 1H), 7.50 (s , 1H), 7.25 (br, 2H), 4.46 (br, 1H), 3.34 (br, 1H), 2.82 (s, 3H), 2.81 (s, 3H), 2.26-2.06 (m, 3H), 2.15 ( s, 3H), 1.91-1.85 (m, 4H), 1.83 (s, 3H), 1.80 (s, 3H).

Example 30

(2- (5-chloro-2-((5-methyl-1- (4-hydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl ) Aminophenyl) dimethylphosphine sulfide

Compound E12 (100 mg, 0.15 mmol) and Lawesson's reagent (122 mg, 0.3 mmol) were dissolved in toluene (5 mL) at room temperature, and the reaction solution was heated to 110 ° C and stirred for 4 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and a yellow solid precipitated. The suspension was filtered. The filter cake was washed with toluene (5 mL). The filtrate was concentrated under reduced pressure. The residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E30 (trifluoroacetate salt, 20.7 mg, yield: 29.1%). Properties: white solid.

HPLC: 97.38%@214nm, 97.75%@254nm

LCMS: MS / m (zI): 477.3 [M + H]

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.20–7.66 (m, 2H), 7.46 (brs, 2H), 4.40 (brs, 1H), 4.06 (dd, J = 11.4, 4.1 Hz, 2H), 3.58 (t, J = 11.6 Hz, 2H), 2.20 (brs, 5H), 2.06 (s, 3H), 2.03 (s, 3H), 1.83 (brs, 2H).

Example 31

(2-((5-cyclopropyl-2-((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethylphosphine oxide

At room temperature, compound D51 (50 mg, 0.15 mmol), compound D29 (28 mg, 0.15 mmol) and NH ₄ Cl (24 mg, 0.45 mmol) were dissolved in EtOH (5 mL), and then the reaction solution was heated to 100 ° C. and stirred for 2 hour. The reaction was checked by LCMS, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E31 (hydrochloride, 15mg, yield: 12%), properties: white solid;

HPLC: 98.731%@214nm, 98.291%@254nm

LCMS: MS / m (zI): 467.4 [M + H]

¹ H NMR (400 MHz, MeOD-d ₄ ) δ: 8.64 (br, 1H), 7.70-7.64 (m, 2H), 7.57 (s, 1H), 7.57-7.40 (m, 1H), 7.39-7.35 (m , 1H), 4.47-4.45 (m, 1H), 4.10-4.06 (m, 2H), 3.64-3.57 (m, 2H), 2.26-2.17 (m, 5H), 1.92-1.88 (m, 2H), 1.91 (s, 3H), 1.88 (s, 3H), 1.76-1.72 (m, 1H), 1.12-1.07 (m, 2H), 0.71-0.67 (m, 2H).

Example 32

(2-((5-chloro-2-((5-chloro-1- (2-azazol [3.5] non-7-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Radical) amino) phenyl) dimethyl phosphine oxidation Thing

Trifluoroacetic acid (300 mg, 2.42 mmol) was added to a solution of compound D56 (300 mg, 0.48 mmol) in methylene chloride (10 mL) at room temperature, and the mixture was stirred at room temperature for 1 hour. After the reaction is completed, the reaction solution is concentrated under reduced pressure, and one third is used for preparative high performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E32 (trifluoroacetate salt, 40 mg, yield: 25%, white solid).

HPLC: 95.34%@233nm, 95.85%@254nm

LCMS: MS / m / z (ESI): 520.3 [M + H]

¹ H NMR (400 MHz, MeOD) δ: 8.30 (br, 1H), 8.10 (br, 1H), 7.69-7.64 (m, 2H), 7.53 (br, 1H), 7.35 (t, J = 7.2 Hz, 1H) ), 4.39-4.30 (m, 1H), 3.96 (s, 2H), 3.82 (s, 2H), 2.27-2.23 (m, 2H), 1.94-1.85 (m, 4H), 1.85 (s, 3H), 1.81 (s, 3H), 1.79-1.73 (m, 2H).

Example 33

4-((2- (dimethylphosphoryl) phenyl) amino) -2-((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazole-4 -Yl) amino) pyrimidine-5-carboxylic acid ethyl ester

At room temperature, compound D57 (710 mg, 2.0 mmol), intermediate D29 (400 mg, 2.2 mmol), ammonium chloride (1.1 g, 20 mmol) and ethanol (20 mL) were added to a 100 mL three-necked flask, respectively. The mixture was heated to reflux and stirred for 16 hours. After the reaction was completed, the reaction liquid was cooled to room temperature, and then the reaction liquid was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent gradient DCM / methanol = 80: 1-40: 1) to obtain the target product E33 (600 mg, yield: 60%, white solid).

HPLC: 96.12%@214nm, 97.10%@254nm

LCMS: MS / m (zI): 499.4 [M + H]

¹ H NMR (400MHz, MeOD) δ: 8.71-8.65 (m, 1H), 7.79-7.51 (m, 3H), 7.46-7.31 (m, 2H), 4.36 (q, J = 7.2Hz, 2H), 4.23 (br, 1H), 4.07-4.02 (m, 2H), 3.58-3.52 (m, 2H), 2.17-2.05 (m, 2H), 2.05 (s, 3H), 1.78-1.72 (m, 8H), 1.40 (t, J = 7.2 Hz, 3H).

Example 34

(2-((5-chloro-2-((5-methyl-1- (1-methylpiperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) Amino) phenyl) dimethylphosphine oxide

At room temperature, 1-methyl-4- (5-methyl-4-amino-1H-pyrazol-1-yl) piperidine (100 mg, 0.32 mmol), (2-((2,5-dichloro Pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (120 mg, 0.64 mmol) and trifluoroacetic acid (2 mL) were dissolved in ethanol (3 mL). The reaction solution was heated to 100 ° C, and the reaction was stirred for 12 hours. The reaction was completed by LCMS. The reaction solution was cooled and concentrated under reduced pressure. The residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E34 (3mg, Yield: 1.9%, white solid).

HPLC: 87.09%@214nm, 86.80%@254nm

LCMS: MS / m (zI): 474.4 [M + H]

¹ H NMR (400 MHz, MeOD) δ: 8.39 (br, 1H), 7.96 (brs, 1H), 7.57-7.52 (m, 1H), 7.45 (brs, 2H), 7.22-7.17 (m, 1H), 4.13 -4.10 (m, 1H), 3.01-2.98 (m, 2H), 2.33 (s, 3H), 2.26-2.24 (m, 4H), 2.16 (s, 3H), 1.92-1.90 (m, 2H), 1.86 (s, 3H), 1.82 (s, 3H).

Example 35

(S)-(2-((5-chloro-2-((5-methyl-1- (tetrahydrofuran-3-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino ) Phenyl) dimethyl phosphine oxide

At room temperature, (S) -5-methyl-4-amino-1- (tetrahydrofuran-3-yl) -1H-pyrazole (50.0 mg, 0.3 mmol), (2-((2,5-dichloro Pyrimidine-4-yl) amino) phenyl) dimethylphosphine oxide (104.0 mg, 0.33 mmol) was dissolved in isopropanol (2 mL), then trifluoroacetic acid (680.0 mg, 6.0 mmol) was added, and the mixture was heated The reaction was stirred at 120 ° C for 4 minutes, and the reaction was completed by LCMS. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E35 (35.0 mg, yield: 26%, white solid).

HPLC: 98.44%@214nm, 99.37%@254nm

LCMS: Rt: 1.1.3min; MS m / z (ESI): 447.4 [M + H]

¹ H NMR (400 MHz, DMSO) δ 11.19 (s, 1H), 8.51 (brs, 1H), 8.06 (s, 1H), 7.54 (dd, J = 12.0 Hz, 8.0 Hz, 1H), 7.45 (s, 1H), 7.31 (br, 1H), 7.12-7.09 (m, 1H), 5.05-4.89 (m, 1H), 4.12-3.94 (m, 2H), 3.82-3.80 (m, 2H), 2.30-2.27 ( m, 2H), 2.14 (s, 3H), 1.79 (s, 3H), 1.75 (s, 3H).

Example 36

4-((5-chloro-4-((2- (dimethylphosphino) phenyl) amino) pyrimidin-2-yl) amino) -1- (tetrahydro-2H-pyran-4-yl ) -1H-pyrazole-5-carbonitrile

At room temperature, 5-cyano-4-amino-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazole (150 mg, 0.78 mmol) and (2-((2,5-di Chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (250 mg, 0.78 mmol) was dissolved in ethanol (10 mL), and then ammonium chloride (530 mg, 10 mmol) was added to the solution; the reaction system was in argon It was heated to 80 ° C under the protection of gas, and the reaction was stirred overnight. After the reaction was completed, the reaction solution was cooled to room temperature, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to give the final product E36 (trifluoroacetate). The product was dissolved in dichloromethane / methanol (20 / 1,5mL), the solution was washed with saturated aqueous sodium bicarbonate (2mL × 3) and water (2mL × 2), and the residue was dried to obtain the free target product E36 (40mg) , Yield 11%, white solid)

HPLC: 97.21%@214nm, 96.07%@254nm, Rt: 8.807min.

LCMS: MS / m (zI): 472.4 [M + H], Rt: 7.395min.

¹ H NMR (400MHz, DMSO-d6) δ: 11.23 (brs, 1H), 9.64 (s, 1H), 8.51 (br, 1H), 8.19 (s, 1H), 7.83 (s, 1H), 7.63-7.57 (m, 1H), 7.47-7.44 (m, 1H), 7.20-7.17 (m, 1H), 4.57-7.53 (m, 1H), 4.00-3.96 (m, 2H), 3.54-3.47 (m, 2H) , 2.05-1.90 (m, 4H), 1.80 (s, 3H), 1.77 (s, 3H).

Example 37

(2- (5-cyclopropyl-2- (5-methyl- (tetrahydropyran-4-2-yl) -pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl ) Dimethylphosphine oxide

At room temperature, (2-((2-chloro-5-isopropylpyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (30 mg, 0.09 mmol), and 5-methyl-1- (Tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-amine (17 mg, 0.09 mmol) and ammonium chloride (24 mg, 0.45 mmol) were dissolved in ethanol (5 mL), and the reaction mixture was heated The reaction was stirred at 100 ° C for 10 hours. LCMS detected the reaction was complete. The reaction solution was cooled to room temperature and filtered, the filtrate was concentrated, and the residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E37 (hydrochloride, 8.1 mg, yield: 12%, yellow solid);

HPLC: 98.624%@214nm, 98.489%@254nm

LCMS: MS / m (zI): 469.5 [M + H]

¹ H NMR (400 MHz, MeOD-d ₄ ) δ: 8.73 (br, 1H), 7.70-7.64 (m, 2H), 7.65-7.55 (m, 2H), 7.36 (br, 1H), 4.46-4.40 (m , 1H), 4.11-4.06 (m, 2H), 3.64-3.57 (m, 2H), 3.16-3.12 (m, 1H), 2.26-2.17 (m, 5H), 1.91-1.87 (m, 8H), 1.31 (d, J = 6.4Hz, 6H).

Example 38

(2-((2-((1-tert-butyl-1H-pyrazol-4-yl) amino) -5-chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide

At room temperature, (2-((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (230 mg, 0.7 mmol), 1-tert-butyl-1H-pyrazole- 4-amine (100 mg, 0.7 mmol) and ammonium chloride were dissolved in ethanol (3 mL). The reaction solution was heated to 85 ° C and stirred for 12 hours. LCMS detected the reaction was complete, the reaction solution was cooled and filtered, and the filtrate was subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure target product E38 (170mg, Yield: 58%, light red solid)

HPLC: 99.31%@214nm, 99.56%@254nm.

LCMS: Rt: 1.327 min; MS m / z (ESI): 419.4 [M + H].

¹ H NMR (400MHz, DMSO) δ 11.11-10.82 (m, 1H), 9.21 (s, 1H), 8.15 (s, 1H), 7.75 (br, 1H), 7.61 (dd, J = 16.0Hz, 8.0 Hz, 1H), 7.53-7.45 (m, 2H), 7.19 (t, J = 8.0Hz, 1H), 1.80 (s, 3H), 1.76 (s, 3), 1.45 (s, 9H).

Example 39

(2-((5-Bromo-2-((1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl ) Dimethylphosphine oxide

At room temperature, 1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-amine (80 mg, 0.48 mmol) was dissolved in ethanol (10 mL), then (2-((5- Bromo-2-chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (173 mg, 0.48 mol) and ammonium chloride (260 mg, 4.8 mmol). The reaction solution was heated to 80 ° C and stirred overnight. The reaction was detected by LCMS. The reaction solution was cooled and filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to high-performance liquid chromatography (removal gradient:

) Purification to obtain product E39 (25 mg, yield: 20%, off-white solid).

HPLC: 98.296%@214nm, 98.785%@254nm

LCMS: MS / m (zI): 491.3 [M + H]

1H NMR (400MHz, DMSO) δ 11.20-10.20 (m, 1H), 9.27 (brs, 1H), 8.79-7.90 (m, 2H), 7.63-7.54 (m, 3H), 7.41 (br, 1H), 7.24 (br, 1H), 4.21 (br, 1H), 3.95-3.93 (m, 2H), 3.45-3.42 (m, 2H), 1.86 (br, 2H), 1.77-1.74 (m, 8H).

Example 40

1- (4-((5-chloro-4-((2- (dimethylphosphoryl) phenyl) amino) pyrimidin-2-yl) amino) -5-methyl-1H-pyrazole-1- Yl) piperidin-1-yl) -2-hydroxy-1-one

At room temperature, convert (2-((5-chloro-2-((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) Amino) phenyl) dimethylphosphine oxide (100mg, 0.2mmol), glycolic acid (30mg, 0.4mmol), HATU (114mg, 0.3mmol) and diisopropylethylamine (72mg, 0.6mmol) dissolved in N , N-dimethylformamide (2mL). The reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was diluted with water (5 mL), and then extracted with dichloromethane / methanol (10: 1, 10 mL × 3). The organic phases were combined, dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. HPLC preparative chromatography (eluent gradient:

) Purification to obtain pure product E40 (15 mg, Yield: 15%, white solid).

HPLC: 97.85%@214nm, 97.82%@254nm

LCMS: MS / m / z (ESI): 518.5 [M + H]

¹ H NMR (400MHz, DMSO) δ 11.18 (s, 1H), 8.49 (brs, 2H), 8.06 (s, 1H), 7.60-7.50 (m, 1H), 7.45 (s, 1H), 7.32 (brs , 1H), 7.11 (t, J = 8.0Hz, 1H), 4.58-4.55 (m, 1H), 4.51-4.35 (m, 2H), 4.22-4.06 (m, 2H), 3.83-3.81 (m, 1H ), 3.14-3.12 (m, 1H), 2.80-2.77 (m, 1H), 2.16 (s, 3H), 2.02-1.81 (m, 4H), 1.79 (s, 3H), 1.75 (s, 3H).

¹⁹ F NMR (376.5MHz, DMSO) δ-73.63, -75.51.

Example 41

2-((5-chloro-2-((1- (2,2-difluoroethyl) piperidin-4-yl) -5-methyl-1H-pyrazol-4-yl) amino) pyrimidine- 4-yl) amino) phenyl) dimethylphosphine oxide

At room temperature, convert (2-((5-chloro-2-((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) Amino) phenyl) dimethylphosphine oxide (100mg, 0.2mmol), 1,1-difluoro-2-iodoethane (58mg, 0.3mmol) and diisopropylethylamine (72mg, 0.6mmol) were dissolved In N, N-dimethylformamide (2mL). The reaction solution was heated to 70 ° C and stirred for 12 hours. The reaction was completed by LCMS, the reaction mixture was cooled and filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to high performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E41 (formate, 15 mg, yield: 15%, white solid).

HPLC: 94.86%@214nm, 96.38%@254nm

LCMS: MS / m / z (ESI): 524.3 [M + H]

¹ H NMR (400 MHz, MeOD) δ 8.42 (brs, 1H), 8.17 (brs, 1H), 7.96 (s, 1H), 7.55 (dd, J = 16.0 Hz, 8.0 Hz, 1H), 7.46 (s, 1H), 7.46 (br, 1H), 7.19 (t, J = 8.0Hz, 1H), 6.01 (tt, J = 56.0Hz, 4.0Hz, 1H), 4.17-4.11 (m, 1H), 3.16-3.13 ( m, 2H), 2.90-2.82 (m, 2H), 2.53-2.47 (m, 2H), 2.26-2.22 (m, 2H), 2.16 (s, 3H), 1.89-1.80 (m, 8H).

¹⁹ F NMR (376.5MHz, MeOD) δ-120.55

Example 42

2- (4- (4-((5-chloro-4-((2- (dimethylphosphoryl) phenyl) amino) pyrimidin-2-yl) amino) -5-methyl-1H-pyrazole -1-yl) piperidin-1-yl) acetic acid formate

At room temperature, methyl 2- (4- (4-((5-chloro-4-((2- (dimethylphosphoryl) phenyl) amino) pyrimidin-2-yl) amino) -5-methyl Yl-1H-pyrazol-1-yl) piperidin-1-yl) acetate (50 mg, 0.096 mmol) and lithium hydroxide (7 mg, 0.29 mmol) were dissolved in methanol (2 mL) and water (1 mL). The reaction solution was stirred at room temperature for 12 hours. LCMS detected that the reaction was complete, the reaction solution was filtered, and the filtrate was directly passed through high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain the pure target product E42 (formate, 40mg, Yield: 80%, colorless oily liquid)

HPLC: 97.90%@214nm, 98.25%@254nm.

LCMS: Rt: 0.83 min; MS m / z (ESI): 518.4 [M + H].

¹ H NMR (400MHz, MeOD) δ 8.58 (br, 1H), 8.25-7.95 (m, 1H), 7.71 (br, 2H), 7.56 (s, 1H), 7.43 (br, 1H), 4.66 (br , 1H), 4.28-7.19 (m, 2H), 3.86-3.83 (m, 2H), 3.64-3.38 (m, 2H), 2.55-2.50 (m, 2H), 2.50 2.10 (m, 5H), 1.93 1.89 (m, 6H).

Example 43

4-((2- (dimethylphosphoryl) phenyl) amino) -2-((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazole-4 -Yl) amino) pyrimidine-5-carbonitrile

At room temperature, (2-((5-bromo-2-((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine -4-yl) amino) phenyl) dimethylphosphine oxide (100 mg, 0.19 mmol), zinc cyanide (22 mg, 0.19 mmol), tris (dibenzylideneacetone) dipalladium (17.3 mg, 0.02 mmol) It was mixed with bis (diphenylphosphine) ferrocene (10.5 mg, 0.02 mmol) and added to N, N-dimethylformamide (5 mL). The reaction system was heated to 100 ° C. under a nitrogen atmosphere and stirred for 8 hours. After the reaction was checked by LCMS, the reaction solution was cooled to room temperature, the mixture was diluted with water (20 mL), and then extracted with dichloromethane (30 mL × 3). The organic phases were combined, dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. Preparative chromatography using high performance liquid chromatography (eluent gradient:

) Purification to obtain product E43 (20 mg, Yield: 23.5%, white solid)

HPLC: 99.068%@214nm, 99.421%@254nm

LCMS: MS / m (zI): 452.4 [M + H]

¹ H NMR (400 MHz, MeOD-d ₄ ): δ 8.50-8.07 (m, 2H), 7.61-7.23 (m, 4H), 4.37-4.33 (m, 1H), 4.09-4.04 (m, 2H), 3.61-3.54 (m, 2H), 2.25-2.15 (m, 2H), 2.15 (s, 3H), 1.87-1.78 (m, 2H), 1.82 (s, 3H), 1.79 (s, 3H).

Example 44

2-((5-chloro-2-((5-chloro-1- (2-methyl-2-azo [3.5] nonyl-7-yl) -1H-pyrazol-4-yl) amino) Pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide

(2-((5-chloro-2-((5-chloro-1- (2-azazol [3.5] nonane-7-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethylphosphine oxide (MED-SIM-2-157) (200 mg, 0.38 mmol) and formaldehyde aqueous solution (100 mg, 37% aqueous solution, 1.2 mmol) in methanol (20 mL) at room temperature The reaction was stirred for 16 hours, and then sodium triacetate borohydride (270 mg, 1.2 mmol) was added, and the reaction solution was stirred at room temperature for 30 minutes. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was dispersed in water (10 mL) and dichloromethane (10 mL). After the two phases were separated, the aqueous phase was extracted with dichloromethane (20 mL × 3). The organic phases are combined, dried and filtered with anhydrous sodium sulfate, the filtrate is concentrated under reduced pressure, and the residue is subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E44 (the resulting TFA salt is free with potassium carbonate) (50mg, Yield: 23%, white solid)

HPLC: 97.66%@233nm, 98.87%@254nm

LCMS: MS / m (zI): 534.4 [M + H]

¹ H NMR (400MHz, MeOD) δ: 8.36 (br, 1H), 8.01 (s, 1H), 7.63 (m, 2H), 7.48-7.46 (m, 1H), 7.24-7.20 (m, 1H), 4.30 -4.23 (m, 1H), 3.28 (s, 2H), 3.13 (s, 2H), 2.42 (s, 3H), 2.12-2.08 (m, 2H), 1.93-1.82 (m, 4H), 1.85 (s , 3H), 1.82 (s, 3H), 1.67-1.61 (m, 2H).

Example 45

(2- (5-Bromo-2-((1- (2-methoxyethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine Oxide

At room temperature, combine 1- (2-methoxyethyl) -1H-pyrazol-4-amine (47.0 mg, 0.33 mmol) and (2-((5-Amino-2-chloropyrimidin-4-yl) Amino) phenyl) dimethyl phosphine oxide (100.0 mg, 0.28 mmol) was dissolved in isopropanol (5 mL), and then trifluoroacetic acid (319.2 mg, 2.80 mmol) was added to the reaction system; the reaction solution was heated to The reaction was stirred at 120 ° C for 3 hours. After the reaction, the reaction solution was cooled and filtered, the filter cake was washed with isopropanol (20 ml), the filtrate was concentrated under reduced pressure, and the residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E45 (29.9 mg, Yield: 23%, brown solid).

HPLC: 99.62%@214nm, 99.78%@254nm

LCMS: MS / m (zI): 467.2 [M + H]

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 9.26 (brs, 1H), 8.59-8.21 (m, 2H), 7.63-7.55 (m, 3H), 7.41 (brs, 1H), 7.22 (brs, 1H), 4.12 (brs, 2H), 3.61 (brs, 2H), 3.32 (s, 3H), 1.79 (s, 3H), 1.76 (s, 3H).

Example 46

(2-((5-Bromo-2-((1- (1- (2-fluoroethyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) Amino) phenyl) dimethylphosphine oxide

At room temperature, convert (2-((5-bromo-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine oxide hydrochloride (50mg, 0.095mmol) and N, N-diisopropylethylamine (49mg, 0.38mmol) were dissolved in N, N-dimethylformamide (15mL) 1-Fluoro-2-iodoethane (25mg, 0.143mmol) was added dropwise to the reaction system; the reaction was heated to 70 ° C and stirred for 16 hours. LCMS detected the reaction was complete. The reaction solution was cooled to room temperature and diluted with water (50 mL). The mixture was extracted with ethyl acetate (20 mL × 4). The organic phases were combined and washed with saturated sodium chloride solution (50 mL × 2), then dried. Sodium sulfate was dried and filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain the target product E46 (4.3 mg, yield: 7.9%, white solid).

HPLC: 95.2%@214nm, 95.8%@254nm

LCMS: MS / m (zI): 536.5 [M + H]

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 8.14 (brs, 2H), 7.73-7.63 (m, 3H), 7.41 (br, 2H), 4.68-4.64 (m, 1H), 4.56-4.52 (m , 1H), 3.99-3.95 (m, 1H), 3.11-3.07 (m, 2H), 2.81-2.78 (m, 1H), 2.74-2.71 (m, 1H), 2.33-2.25 (m, 2H), 1.96 (br, 4H), 1.83 (s, 3H), 1.80 (s, 3H).

¹⁹ F NMR (376.5MHz, MeOH-d ₄ ) δ-200.00.

Example 47

3- (4-((5-chloro-4-((2- (dimethylphosphoryl) phenyl) amino) pyrimidin-2-yl) amino) -5-methyl-1H-pyrazole-1- Yl) piperidin-1-yl) propionitrile

At room temperature, convert (2-((5-chloro-2-((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) Amino) phenyl) dimethylphosphine oxide (72mg, 0.16mmol) was dissolved in N, N, -dimethylformamide (10mL), and then acrylonitrile (10mg, 0.16mol) and sodium carbonate (34mg, 0.32 mmol); the reaction solution was stirred at 65 ° C. overnight. The reaction was checked by LCMS. The reaction solution was cooled and filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to high-performance liquid chromatography (removal gradient:

) Purification to obtain product E47 (25 mg, yield: 20%, off-white solid).

HPLC: 93.920%@214nm, 94.278%@254nm

LCMS: MS / m (zI): 513.4 [M + H]

¹ H NMR (400MHz, MeOD) δ 8.42 (brs, 1H), 7.96 (s, 1H), 7.58-7.40 (m, 3H), 7.20-7.17 (m, 1H), 4.16-4.09 (m, 1H) , 3.11-3.08 (m, 2H), 2.75-2.60 (m, 4H), 2.31-2.17 (m, 4H), 2.16 (s, 3H), 1.90-1.88 (m, 2H), 1.86 (s, 3H) , 1.82 (s, 3H).

Example 48

(2-((5-chloro-2-((5-methyl-1- (1- (2- (methylsulfonyl) ethyl) piperidin-4-yl) -1H-pyrazole-4- Yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine oxide

At room temperature, convert (2-((5-chloro-2-((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) Amino) phenyl) dimethylphosphine oxide (57mg, 0.124mmol) was dissolved in N, N, -dimethylformamide (5mL), and then added methyl vinyl sulfone (13mg, 0.124mol) and sodium carbonate (26 mg, 0.248 mmol), the reaction solution was heated to 65 ° C and the reaction was stirred overnight. The reaction was checked by LCMS, the reaction liquid was cooled and filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to high performance liquid preparative chromatography (removal gradient:

) Purification to obtain product E48 (25 mg, yield: 20%, off-white solid).

HPLC: 93.512%@214nm, 96.920%@254nm

LCMS: MS / m (zI): 566.4 [M + H]

¹ H NMR (400 MHz, MeOD) δ 8.43 (br, 1H), 8.17 (br, 1H), 7.96 (br, 1H), 7.56-7.41 (m, 1H), 7.47 (s, 1H), 7.19 (t , J = 8.0Hz, 1H), 4.22-4.15 (m, 1H), 3.36-3.34 (m, 2H), 3.25-3.20 (m, 2H), 3.11 (s, 3H), 2.98-2.96 (m, 2H ), 2.40-2.37 (m, 2H), 2.28-2.19 (m, 2H), 2.17 (s, 3H), 1.95-1.92 (m, 2H), 1.86 (s, 3H), 1.82 (s, 3H).

Example 49

2-((5-Amino-2-((1- (1- (2,2-difluoroethyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Radical) amino) phenyl) dimethyl phosphine oxide

At room temperature, convert (2-((5-bromo-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine oxide hydrochloride (60mg, 0.1mmol) and N, N-diisopropylethylamine (52mg, 0.4mmol) were dissolved in N, N-dimethylformamide (15mL), and then reacted 1,1-Difluoro-2-iodoethane (30 mg, 0.15 mmol) was added dropwise to the system; the reaction solution was heated to 70 ° C. and stirred for 16 hours. The reaction was checked by LCMS. The reaction solution was cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (20 mL × 4), the organic phases were combined, washed with saturated sodium chloride solution (50 mL × 2), and dried over anhydrous sodium sulfate Dry filtration, the filtrate is concentrated under reduced pressure, and the residue is subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain the target product E49 (4.6 mg, yield: 6.8%, Lot # white solid).

HPLC: 93.9%@214nm, 95.5%@254nm

LCMS: MS / m (zI): 554.3 [M + H]

¹ H NMR (400 MHz, MeOH-d ₄ ) δ 8.14-7.80 (m, 2H), 7.77-7.65 (m, 2H), 7.55-7.38 (m, 3H), 6.15-5.86 (m, 1H), 3.93 (br, 1H), 3.09-3.05 (m, 2H), 2.85-2.75 (m, 2H), 2.43-2.35 (m, 2H), 1.91 (br, 4H), 1.83 (s, 3H), 1.80 (s , 3H).

¹⁹ F NMR (376.5MHz, MeOH-d ₄ ) δ-120.82

Example 50

(2-((5-chloro-2-((1- (1- (2-methylsulfonyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl ) Amino) phenyl) dimethylphosphine oxide

(2-((5-chloro-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine oxide (120 mg, 0.27 mmol) was dissolved in N, N, -dimethylformamide (10 mL), then (methylsulfonyl) ethylene (30 mg, 0.27 mol) and sodium carbonate (60 mg, 0.54 mmol) were added . The reaction solution was heated to 60 ° C and stirred overnight. The reaction was detected by LCMS, the reaction solution was filtered, the filtrate was concentrated under reduced pressure, and the residue was prepared by high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E50 (25 mg, yield: 20%). Properties: Off-white solid.

¹ H NMR (400MHz, DMSO) δ 11.5-10.5 (m, 1H), 9.24 (s, 1H), 8.13 (s, 1H), 7.65-7.52 (m, 3H), 7.42 (br, 1H), 7.22 (br, 1H), 3.99 (br, 1H), 3.30-3.29 (m, 2H), 3.04 (s, 3H), 3.01-2.98 (m, 2H), 2.75-2.73 (m, 2H), 2.15-2.10 (m, 2H), 1.91-1.75 (m, 4H), 1.78 (s, 3H), 1.75 (s, 3H).

HPLC: 96.947%@214nm, 97.696%@254nm

LCMS: MS / m (zI): 552.4 [M + H]

Example 51

(2-((5-chloro-2-((1- (1- (2-methoxyethyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Group) amino) phenyl) dimethyl phosphine oxide

(2-((5-chloro-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine oxide (100 mg, 0.22 mmol), 1-bromo-2-methoxyethane (31 mg, 0.22 mmol), and N, N-diisopropylethylamine (28 mg, 0.22 mmol) dissolved in N, N -Dimethylformamide (3 mL). The reaction solution was stirred at room temperature for 10 hours, and LCMS detected that the reaction was complete. The reaction solution was directly prepared by high performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E51 (25.5 mg, yield: 23%), character: white solid.

¹ H NMR (400 MHz, MeOD) δ: 8.41-8.05 (m, 1H), 8.05 (s, 1H), 7.71-7.61 (m, 3H), 7.43 (s, 1H), 7.35 (br, 1H), 3.97 -3.93 (m, 1H), 3.58-3.55 (m, 2H), 3.36 (s, 3H), 3.10-3.06 (m, 2H), 2.65-2.61 (m, 2H), 2.27-2.21 (m, 2H) , 1.98-1.88 (m, 4H), 1.84 (s, 3H), 1.81 (s, 3H).

HPLC: 97.049%@214nm, 98.876%@254nm

LCMS: MS / m (zI): 504.4 [M + H]

Example 52

(2-((5-chloro-2-((5-chloro-1- (1- (2- (methylsulfonyl) ethyl) piperidin-4-yl) -1H-pyrazol-4-yl ) Amino) pyrimidin-4-yl) amino) phenyl) dimethyloxy Phosphine

At room temperature, place (2-((5-chloro-2-((5-chloro-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino ) Phenyl) dimethyl phosphine oxide (90mg, 0.19mmol), methanesulfonyl ethylene (24mg, 0.23mmol) and diisopropylethylamine (49mg, 0.38mmol) dissolved in N'N-dimethyl methyl Amide (2mL). The reaction solution was stirred at 60 ° C for 2 hours. LCMS detected the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the residue was subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure product E52 (45mg, yield: 35%), character: beige solid

¹ H NMR (400MHz, MeOD) δ 8.37 (brs, 1H), 8.01 (s, 1H), 7.65 (s, 1H), 7.58 (dd, J = 12.0Hz, 8.0Hz, 1H), 7.50-7.46 ( m, 1H), 7.22 (t, J = 8.0 Hz, 1H), 4.38-4.28 (m, 1H), 3.33-3.30 (m, 2H), 3.14-3.10 (m, 2H), 3.10 (s, 3H) , 2.92-2.90 (m, 2H), 2.31-2.11 (m, 4H), 1.94-1.91 (m, 2H), 1.86 (s, 3H), 1.82 (s, 3H).

HPLC: 99.83%@214nm, 99.96%@254nm

LCMS: Rt: 0.96 min; MS m / z (ESI): 586.4 [M + H].

Example 53

2-((5-chloro-2-((1- (2- (2-methoxyethyl) -2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl ) Amino) pyrimidin-4-yl) amino) phenyl) dimethyloxy Phosphine

At room temperature, 2- (2-methoxyethyl) -7- (4-amino-1H-pyrazol-1-yl) -2-azaspiro [3.5] nonane (100 mg, 0.37 mmol), (2-((2,5-Dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (120mg, 0.37mmol) and ammonium chloride (196mg, 3.7mmol) were dissolved in ethanol (5mL) During the reaction, the reaction system was heated to reflux and the reaction was stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target pure product E53 (30.9mg, yield: 15.4%), properties: white solid

¹ H NMR (400MHz, DMSO-d6): δ 11.30-10.75 (m, 1H), 9.24 (s, 1H), 8.85-8.13 (m, 2H), 7.92-6.44 (m, 5H), 3.94 (brs , 1H), 3.29-3.27 (m, 2H), 3.22 (s, 3H), 2.97 (s, 2H), 2.89 (s, 2H), 2.56-2.54 (m, 2H), 1.93-1.90 (m, 2H ), 1.82-1.75 (m, 2H), 1.79 (s, 3H), 1.75 (s, 3H), 1.68-1.39 (m, 4H).

HPLC: 97.47%@214nm, 98.72%@254nm

LCMS: MS / m (zI): 544.4 [M + H]

Example 54

(2-((5-chloro-2-((1- (2-methyl-2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethylphosphine oxide

At room temperature, (2-((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (3) (80 mg, 0.25 mmol), 1- (2-methyl- 2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-amine (8) (56mg, 0.25mol) and ammonium chloride (132.5mg, 2.5mmol) were dissolved in ethanol (5mL) Then, the reaction solution was heated to 80 ° C and the reaction was stirred overnight. After the reaction was completed, the reaction solution was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure, and the residue was prepared by high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target pure product E54 (11mg, yield: 8.8%), properties: white solid

¹ H NMR (400 MHz, MeOD) δ: 8.45 (br, 1H), 8.06 (s, 1H), 7.69-7.35 (m, 5H), 3.90 (brs, 1H), 3.22 (s, 2H), 3.12 (s , 2H), 2.41 (s, 3H), 2.08-2.05 (m, 2H), 1.90-1.85 (m, 2H), 1.81 (s, 3H), 1.75 (s, 3H), 1.68-.58 (m, 4H).

HPLC: 97.44%@214nm, 98.29%@254nm

LCMS: MS / m (zI): 500.2 [M + H]

Example 55

2-((5-chloro-2-((1- (2- (2-fluoroethyl) -2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) amino ) Pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide

At room temperature, 2- (2-fluoroethyl) -7- (4-amino-1H-pyrazol-1-yl) -2-azaspiro [3.5] nonane (360mg, 1.43mmol), (2 -((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (450 mg, 1.43 mmol) and trifluoroacetic acid (1600 mg, 14.3 mmol) dissolved in isopropanol (10 mL) In the process, the reaction system was heated to reflux, and the reaction was stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification, to obtain the target pure product E55 (trifluoroacetate, 62.2mg, yield: 8.1%), properties: white solid

¹ H NMR (400MHz, MeOD): δ 8.09 (brs, 2H), 7.93-6.79 (m, 5H), 4.80-4.75 (m, 1H), 4.69-4.63 (m, 1H), 4.23 (brs, 1H ), 4.06-4.00 (m, 4H), 3.73-3.56 (m, 2H), 2.27-2.16 (m, 2H), 1.99 (br, 2H), 1.86 (s, 3H), 1.82 (s, 3H), 1.86-1.80 (m, 4H).

¹⁹ F NMR (376.5MHz, MeOD): δ-77.14, -218.41

HPLC: 95.27%@214nm, 97.41%@254nm

LCMS: MS / m (zI): 532.4 [M + H]

Example 56

(2-((5-chloro-2-((1-methylcyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino ) Phenyl) dimethyl phosphine oxide

At room temperature, 2-methyl-5- (4-amino-1H-pyrazol-1-yl) cyclopentyl [c] pyrrolidine (430 mg, 2.07 mmol), (2-((2,5- Dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (650 mg, 2.07 mmol) was dissolved in ethanol (10 mL), and ammonium chloride (1.3 g, 20.7 mmol) was added; the reaction solution was heated to The reaction was stirred overnight at 80 ° C. LCMS detected the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain the target product E56 (103.7 mg, yield: 10.6%). Properties: Off-white solid.

¹ H NMR (400 MHz, MeOD) δ 8.40-8.05 (m, 2H), 7.72-7.62 (m, 3H), 7.41-7.36 (m, 2H), 4.64 (brs, 1H), 2.81 (d, J = 5.2Hz, 4H), 2.36 (s, 3H), 2.26 (d, J = 4.0Hz, 2H), 2.08 (br, 2H), 1.91 (br, 2H), 1.85 (s, 3H), 1.81 (s, 3H).

HPLC: 94.994%@214nm, 96.542%@254nm

LCMS: MS / m (zI): 486.4 [M + H]

Example 57

(2-((5-chloro-2-((1- (1- (2-hydroxy-2-methylpropyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine -4-yl) amino) phenyl) dimethylphosphine sulfide Thing

At room temperature, (2-((5-chloro-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine sulfide (310mg, 0.62mmol), 2,2-dimethylethylene oxide (447mg, 6.2mmol) and potassium carbonate (427mg, 3.1mmol) dissolved in ethanol / water (6mL / 1.5mL) in. The reaction solution was heated to 110 ° C in a microwave reactor, and the reaction was stirred for 20 minutes. LCMS detected the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the residue was subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure product E57 (30mg, yield: 21%), properties: white solid

¹ H NMR (400MHz, DMSO) δ 9.39-9.27 (m, 2H), 8.16 (brs, 1H), 7.94 (brs, 1H), 7.72 (brs, 2H), 7.48 (brs, 1H), 7.22 (brs , 2H), 4.14 (s, 1H), 3.74 (brs, 1H), 3.33 (br, 2H), 3.03-3.00 (m, 2H), 2.23 (s, 2H), 2.22 (br, 2H), 1.99 ( s, 3H), 1.95 (s, 3H), 1.69 (brs, 4H), 1.13 (s, 6H).

HPLC: 99.62%@214nm, 99.73%@254nm

LCMS: Rt: 1.16 min; MS m / z (ESI): 534.5 [M + H].

Example 58

(2-((5-chloro-2-((1- (2-methyl-2-azaspiro [3.3] hept-6-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethylphosphine oxide

At room temperature, 2-methyl-6- (4-amino-1H-pyrazol-1-yl) -2-azaspiro [3.3] heptane (150 mg, 0.78 mmol), (2-((2, 5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (8) (245 mg, 0.78 mmol) was dissolved in ethanol (10 mL), and ammonium chloride (560 g, 7.8 mmol) was added, The reaction solution was heated to 80 ° C and the reaction was stirred overnight. The reaction was completed by LCMS, the reaction solution was filtered, the filtrate was concentrated under reduced pressure, and the residue was prepared by high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target product E58 (19.4 mg, yield: 20%), properties: off-white solid.

¹ H NMR (400MHz, DMSO) δ 11.35-11.10 (m, 1H), 8.70-8.50 (m, 1H), 8.03-7.74 (m, 3H), 7.67-7.50 (m, 2H), 7.30-7.10 ( m, 2H), 5.05 (brs, 1H), 4.44 (br, 2H), 3.83 (br, 2H), 3.11 (s, 3H), 2.65-2.50 (m, 2H), 1.96 (br, 2H), 1.79 (s, 3H), 1.76 (s, 3H).

HPLC: 95.553%@214nm, 97.271%@254nm

LCMS: MS / m (zI): 472.4 [M + H]

Examples 59 and 60

(2-((5-chloro-2-((1- (3- (4-methylpiperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Radical) amino) phenyl) dimethyl phosphine oxide (Example 59 single isomer 1: Rt: 5.078 min; Example 60 single isomer 2: Rt: 5.106 min)

At room temperature, 1-methyl-4- (3- (4-amino-1H-pyrazol-1-yl) cyclobutyl) piperazine (106 mg, 0.45 mmol), (2-((2,5- Dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (157 mg, 0.5 mmol) and ammonium chloride (240 mg, 4.5 mmol) were dissolved in isopropanol (3 mL). The reaction solution was stirred at 80 ° C for 12 hours. LCMS detected the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the residue was then subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure product E59 (single isomer 1: Rt: 5.078 min) (31.5 mg, yield: 12%), trait: white solid; and

E60 (single isomer 2: Rt: 5.106 min) (4.1 mg, yield: 2%), character: white solid.

E59 (single isomer 1: Rt: 5.078min)

¹ H NMR (400MHz, MeOD): δ 8.25 (br, 1H), 8.06 (s, 1H), 7.80-7.60 (m, 3H), 7.48 (brs, 1H), 7.34 (brs, 1H), 4.69 ( brs, 1H), 3.10-3.05 (m, 1H), 2.72-2.26 (m, 12H), 1.85 (s, 3H), 1.82 (s, 3H).

HPLC: 99.71%@214nm, 99.81%@254nm, Rt: 5.078min

LCMS: Rt: 0.855 min; MS m / z (ESI): 515.4 [M + H].

E60 (single isomer 2: Rt: 5.106min)

¹ H NMR (400MHz, MeOD): δ 8.37 (br, 1H), 8.06 (s, 1H), 7.83-7.54 (m, 3H), 7.48-7.45 (m, 1H), 7.33-7.30 (m, 1H) ), 4.41 (br, 1H), 3.08 (br, 1H), 2.65-2.30 (m, 12H), 1.86 (s, 3H), 1.82 (s, 3H).

HPLC: 96.28%@214nm, 96.20%@254nm, Rt: 5.106min

LCMS: Rt: 0.889 min; MS m / z (ESI): 515.4 [M + H].

Example 61

2-((5-chloro-2-((1- (2-morpholinolinylethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine Oxide

Oxidation of (2-((5-chloro-2-((1- (2-chloroethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine (70mg, 0.16mmol), morphine (27mg, 0.32mmol) and diisopropylethylamine (66mg, 0.48mmol) were dissolved in N, N-dimethylformamide (5mL), the reaction solution was at room temperature The reaction was stirred for 10 hours, and the reaction was completed by LCMS. The reaction solution was filtered, and the filtrate was prepared by high-performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E61 (trifluoroacetate salt, 43.4 mg, yield: 57.1%), character: white solid.

¹ H NMR (400 MHz, MeOD): δ 8.12 (brs, 1H), 7.78-7.40 (m, 6H), 4.49 (brs, 2H), 3.91 (brs, 4H), 3.68-3.64 (m, 2H), 3.37-3.31 (m, 4H), 1.86 (s, 3H), 1.83 (s, 3H).

¹⁹ F NMR (376.5MHz, MeOD): δ-77.18

HPLC: 94.558%@214nm, 94.857%@254nm

LCMS: MS / m (zI): 476.4 [M + H]

Example 62

(2-((5-Bromo-2-((1- (1- (2,2-difluoroethyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethylphosphine sulfide

At room temperature, (2-((5-bromo-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine sulfide (120mg, 0.24mmol), 1,1-difluoro-2-iodoethane (60mg, 0.31mmol) and potassium carbonate (98mg, 0.71mmol) were added to N'N-dimethylformamide (5mL); The reaction solution was heated to 70 ° C, and the reaction was stirred for 24 hours. The reaction was completed by LCMS and the reaction mixture was concentrated. The residue is then subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain E62 (trifluoroacetate salt, 10 mg, yield: 5%), character: white solid.

¹ H NMR (400MHz, MeOD): δ 8.24 (brs, 1H), 7.98-7.41 (m, 6H), 6.57-6.29 (m, 1H), 4.19 (brs, 1H), 3.74-3.69 (m, 4H) ), 3.44-3.33 (m, 2H), 2.24 (brs, 4H), 2.04 (s, 3H), 2.00 (s, 3H).

¹⁹ F NMR (376.5MHz, MeOD): δ-77.12, -122.16.

HPLC: 98.17%@214nm, 98.28%@254nm

LCMS: Rt: 1.30 min; MS m / z (ESI): 572.3 [M + H].

Example 63

2-((5-Bromo-2-((1- (1- (2-hydroxy-2-methylpropyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine- 4-yl) amino) phenyl) dimethylphosphine sulfide

At room temperature, (2-((5-bromo-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine sulfide (85mg, 0.17mmol), 2,2-dimethylethylene oxide (120mg, 1.67mmol) and potassium carbonate (115mg, 0.85mmol) dissolved in ethanol / water (2mL / 0.5mL) Medium; the reaction solution was heated to 110 ° C with a microwave reactor, and the reaction was stirred for 20 minutes. The reaction was completed by LCMS and the reaction mixture was concentrated. The residue is then subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain E63 (trifluoroacetate salt, 10 mg, yield: 6%), character: white solid.

¹ H NMR (400 MHz, MeOD): δ 8.24 (brs, 1H), 7.99-7.40 (m, 6H), 4.18 (brs, 1H), 3.90 (d, J = 11.2 Hz, 1H), 3.53 (brs, 1H), 3.33-3.23 (m, 4H), 2.32 (br, 2H), 3.19 (br, 2H), 2.04 (s, 3H), 2.00 (s, 3H), 1.37 (s, 6H).

¹⁹ F NMR (376.5MHz, MeOD): δ-77.13 (s).

HPLC: 93.70%@214nm, 93.71%@254nm

LCMS: Rt: 1.20 min; MS m / z (ESI): 580.3 [M + H].

Example 64

(2-((5-chloro-2-((1- (2- (pyrrolidin-1-yl) ethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) benzene Radical) dimethyl phosphine oxide

Oxidation of (2-((5-chloro-2-((1- (2-chloroethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine (70 mg, 0.16 mmol), pyrrolidine (6) (35 mg, 0.49 mmol) and diisopropylethylamine (63 mg, 0.49 mmol) were dissolved in N, N-dimethylformamide (5 mL). The reaction solution was stirred at room temperature for 10 hours, and LCMS showed that the reaction was complete. The reaction solution was filtered, and the filtrate was prepared by high-performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E64 (19.3 mg, yield: 26.4%), character: white solid.

¹ H NMR (400MHz, MeOD): δ 8.51-8.05 (m, 2H), 7.68-7.34 (m, 5H), 4.14 (brs, 2H), 2.86 (br, 2H), 2.54 (brs, 4H), 1.85 (s, 3H), 1.81 (s, 3H), 1.85-1.76 (m, 4H).

HPLC: 98.925%@214nm, 98.812%@254nm

LCMS: MS / m (zI): 460.4 [M + H]

Example 65

(2-((5-chloro-2-((1- (1- (2,2-difluoroethyl) piperidin-3-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethylphosphine oxide

At room temperature, (2-((5-chloro-2-((1- (piperidin-3-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine oxide (80mg, 0.18mmol), 1,1-difluoro-2-iodoethane (52mg, 0.28mmol) and potassium carbonate (74mg, 0.54mmol) were added to N'N-dimethyl methyl In amide (4mL), the reaction was stirred at 80 ° C for 16 hours; after the reaction was complete, the reaction solution was cooled to room temperature, the mixture was filtered, and the filtrate was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E65 (trifluoroacetate, 9.3 mg, yield: 10%), character: white solid.

¹ H NMR (400MHz, DMSO-d6) δ: 11.15-10.67 (m, 1H), 9.28 (brs, 1H), 8.33 (br, 1H), 8.14 (s, 1H), 7.63 (br, 1H), 7.57 -7.53 (m, 1H), 7.43 (br, 1H), 7.22 (br, 1H), 6.30-6.02 (m, 1H), 4.11 (br, 1H), 3.10-3.07 (m, 1H), 2.89-2.75 (m, 3H), 2.50-2.40 (m, 1H), 2.27-2.20 (m, 1H), 1.94 (br, 1H), 1.79 (s, 3H), 1.76 (s, 3H), 1.75-1.67 (m , 1H), 1.62-1.55 (m, 2H).

¹⁹ F NMR (376.5MHz, DMSO-d6) δ: -118.78

HPLC: 97.84%@214nm, 99.36%@254nm

LCMS: MS / m (zI): 510.5 [M + H]

Example 66

(2-((5-chloro-2-((1- (2- (2,2-difluoroethyl) -2-azaspiro [3.5] non-7-yl) -1H-pyrazole-4 -Yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyloxy Phosphine

At room temperature, 2- (2,2-difluoroethyl) -7- (4-amino-1H-pyrazol-1-yl) -2-azaspiro [3.5] nonane (30mg, 0.11mmol) , (2-((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (35mg, 0.11mmol) and trifluoroacetic acid (125mg, 1.1mmol) dissolved in isopropanol (5 mL), the reaction system was heated to reflux, and the reaction was stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure, and the residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target pure product E66 (trifluoroacetate, 6.0 mg, yield: 11%), properties: white solid

¹ H NMR (400MHz, MeOD): δ 8.25-8.19 (m, 4H), 7.69-7.67 (m, 2H), 7.36 (brs, 1H), 6.40-5.94 (m, 1H), 4.58 (s, 2H ), 4.24-3.74 (m, 1H), 3.38-3.34 (m, 2H), 2.98 (s, 2H), 2.35-2.05 (m, 4H), 1.91-1.54 (m, 10H).

¹⁹ F NMR (376.5MHz, MeOD): δ-76.13, -123.39

HPLC: 94.41%@214nm, 96.64%@254nm

LCMS: MS / m (zI): 550.4 [M + H]

Example 67

(2-((5-chloro-2-((1- (2-methyl-2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethylphosphine sulfide

At room temperature, (2-((2-((1- (2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) directly) -5-chloropyrimidine-4- Methyl) amino) phenyl) dimethyl phosphine sulfide (60mg, 0.12mmol) and formaldehyde aqueous solution (97mg, 37% aqueous solution, 1.2mmol) in methanol (5mL) solution was added triethylamine (12.1mg, 0.12mmol), stirred After 5 hours of reaction, sodium triacetate borohydride (101.8 mg, 0.48 mmol) was added, and the reaction solution was stirred at room temperature for 2 hours. After the reaction is complete, the reaction solution is concentrated under reduced pressure, and the residue is subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target compound E67 (trifluoroacetate, 20.1mg, yield: 33%), properties: white solid.

¹ H NMR (400 MHz, MeOD): δ 8.15 (brs, 1H), 7.97-7.38 (m, 6H), 4.20 (d, J = 8.0 Hz, 1H), 4.04 (d, J = 8.0 Hz, 1H) , 3.90-3.82 (m, 3H), 2.96 (s, 3H), 2.45-2.09 (m, 2H), 2.04 (s, 3H), 2.00 (3H,) 2.04-1.93 (m, 2H), 1.77-1.64 (m, 4H).

¹⁹ F NMR (376.5MHz, MeOD): δ-77.17

HPLC: 99.64%@214nm, 99.52%@254nm

LCMS: MS / m (zI): 516.1 [M + H]

Example 68

(2-((5-chloro-2-((1- (2- (2,2,2-trifluoroethyl) -2-azaspiro [3.5] human-7-yl) -1H-pyrazole -4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine oxide

At room temperature, (2-((2-((1- (2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) amino) -5-chloropyrimidine-4- Yl) amino) phenyl) dimethylphosphine oxide (48.5 mg, 0.1 mmol), 1,1,1-trifluoro-2-iodoethane (41.8 mg, 0.2 mol) and diisopropylethylamine ( 24.4 mg, 0.2 mmol) was dissolved in acetonitrile (5 mL), the reaction solution was heated to 80 ° C, and the reaction was stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was prepared by high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target pure product E68 (trifluoroacetate salt, 32 mg, yield: 56%), character: white solid.

¹ H NMR (400 MHz, MeOD): δ 8.13 (br, 2H), 7.87-6.96 (m, 5H), 4.40-4.06 (m, 6H), 3.98 (brs, 1H), 2.25 (brs, 2H), 1.99 (brs, 2H), 1.87-1.76 (m, 10H).

¹⁹ F NMR (376.5MHz, MeOD): δ-70.15, -76.84

HPLC: 98.26%@214nm, 98.59%@254nm

LCMS: MS / m (zI): 568.5 [M + H]

Example 69

(2-((5-chloro-2-((1- (2- (2-hydroxy-2-methylpropyl) cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazole- 4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethyl phosphine oxide

At room temperature, (2-((5-chloro-2-((1- (cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Group) amino) phenyl) dimethyl phosphine oxide (160 mg, 0.30 mmol), 2,2-dimethyl ethylene oxide (216 mg, 3.0 mmol) and potassium carbonate (207 mg, 5.0 mmol) dissolved in ethanol / water (2mL / 0.5mL). The reaction solution was heated to 110 ° C in a microwave reactor, and the reaction was stirred for 20 minutes. LCMS detected the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the residue was subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure product E69 (hydrochloride, 30mg, yield: 21%), properties: white solid

¹ H NMR (400MHz, DMSO) δ 9.27 (brs, 1H), 8.14 (s, 1H), 7.63-7.52 (m, 3H), 7.37 (brs, 1H), 7.22 (brs, 1H), 4.72 (brs , 1H), 4.09 (s, 1H), 2.68-2.51 (m, 6H), 2.27 (s, 2H), 2.02 (brs, 2H), 1.85-1.75 (m, 2H), 1.78 (s, 3H), 1.75 (s, 3H), 1.12 (s, 6H).

HPLC: 96.18%@214nm, 97.23%@254nm

LCMS: Rt: 0.96 min; MS m / z (ESI): 544.5 [M + H].

Example 70

(2-((5-chloro-2-((1-ethylcyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino ) Phenyl) dimethyl phosphine oxide

At room temperature, (2-((5-chloro-2-((1- (cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Group) amino) phenyl) dimethyl phosphine oxide (160mg, 0.30mmol), ethyl iodide (56mg, 0.36mmol) and diisopropylethylamine (116mg, 0.90mmol) dissolved in N'N-dimethyl Formamide (1 mL). The reaction solution was stirred at 70 ° C for 12 hours. The reaction was checked by LCMS, and the reaction mixture was concentrated under reduced pressure. The residue is subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure product E70 (14mg, yield: 10%), character: white solid.

¹ H NMR (400MHz, DMSO) δ 9.28 (brs, 1H), 8.14 (s, 1H), 7.65-7.54 (m, 3H), 7.36 (brs, 1H), 7.23 (brs, 1H), 4.67 (brs , 1H), 2.66 (brs, 2H), 2.50-2.40 (m, 6H), 2.02 (brs, 2H), 1.90-1.75 (m, 2H), 1.78 (s, 3H), 1.75 (s, 3H), 1.06 (t, J = 8.0Hz, 3H).

HPLC: 95.05%@214nm, 96.19%@254nm

LCMS: Rt: 0.96 min; MS m / z (ESI): 500.5 [M + H].

Example 71

(2-((5-chloro-2-((1- (2- (2-fluoroethyl) cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino ) Pyrimidin-4-yl) amino) phenyl) dimethyloxy Phosphine

(2-((5-chloro-2-((1- (cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino ) Phenyl) dimethyl phosphine oxide (100 mg, 0.21 mmol), 1-fluoro-2-iodoethane (36 mg, 0.21 mmol) and N, N-isopropylethylamine (54 mg, 0.42 mmol) dissolved in N , N-dimethylformamide (3mL). The reaction solution was stirred at 100 ° C for 8 hours. The reaction was completed by LCMS. The reaction solution was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E71 (25.5 mg, yield: 20.4%), character: white solid.

¹ H NMR (400 MHz, MeOD) δ: 8.41-8.05 (m, 2H), 7.68-7.62 (m, 3H), 7.41-7.35 (m, 2H), 4.65-4.62 (m, 2H), 4.53-4.50 ( m, 1H), 2.90-2.73 (m, 6H), 2.38-2.34 (m, 2H), 2.07-2.06 (m, 2H), 1.94-1.93 (m, 2H), 1.84 (s, 3H), 1.81 ( s, 3H).

¹⁹ H NMR (376.5MHz, MeOD) δ: -221.02

HPLC: 99.172%@214nm, 99.196%@254nm

LCMS: MS / m (zI): 518.4 [M + H]

Example 72

2-((5-chloro-2-((1- (2- (2,2-difluoroethyl) cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl ) Amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine oxide

(2-((5-chloro-2-((1- (cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino ) Phenyl) dimethyl phosphine oxide (150 mg, 0.31 mmol), 1,1-difluoro-2-iodoethane (60 mg, 0.31 mmol) and N, N-isopropylethylamine (54 mg, 0.42 mmol) Soluble in acetonitrile (3mL). The reaction solution was stirred at 70 ° C for 8 hours, and the reaction was completed by LCMS. The reaction solution was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E72 (13.2 mg, yield: 7.8%), character: white solid.

¹ H NMR (400MHz, MeOD): δ 8.10 (br, 2H), 7.70-7.63 (m, 3H), 7.49-7.40 (m, 2H), 6.51-6.15 (m, 1H), 3.90-3.77 (m , 5H), 3.14 (br, 4H), 2.10 (br, 2H), 2.07-2.06 (m, 2H), 1.86 (s, 3H), 1.83 (s, 3H).

¹⁹ H NMR (376.5MHz, MeOD) δ: -123.16

HPLC: 96.968%@214nm, 97.660%@254nm

LCMS: MS / m (zI): 536.5 [M + H]

Example 73

(2-((5-chloro-2-((1- (2- (2,2,2-trifluoroethyl) cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazole- 4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine oxide

(2-((5-chloro-2-((1- (cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino ) Phenyl) dimethyl phosphine oxide (100 mg, 0.21 mmol), 1,1,1-trifluoro-2-iodoethane (48 mg, 0.21 mmol) and N, N-isopropylethylamine (54 mg, 0.42 mmol) was dissolved in acetonitrile (3mL). The reaction solution was stirred at 70 ° C for 8 hours, and the reaction was completed by LCMS. The reaction solution was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E73 (20.2 mg, yield: 20.4%), character: white solid.

¹ H NMR (400 MHz, MeOD) δ: 8.41-8.04 (m, 2H), 7.75-7.60 (m, 3H), 7.40-7.35 (m, 2H), 4.69 (br, 1H), 3.15-3.07 (m, 2H), 2.76-2.72 (m, 4H), 2.64-2.62 (m, 2H), 2.09 (br, 2H), 1.95-1.87 (m, 2H), 1.84 (s, 3H), 1.81 (s, 3H) .

¹⁹ F NMR (376.5MHz, MeOD) δ: -71.05

HPLC: 99.125%@214nm, 99.196%@254nm

LCMS: MS / m (zI): 554.4 [M + H]

Example 74

2-((5-chloro-2-((1- (2- (2-hydroxy-2-methylpropyl) -2-azaspiro [3.5] human-7-yl) -1H-pyrazole- 4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine sulfide

Under the protection of argon, add (2-((2-((1- (2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) directly to the microwave tube) -5-chloropyrimidin-4-yl) amino) phenyl) dimethyl phosphine sulfide (60 mg, 0.12 mmol), 2,2-dimethyl ethylene oxide (86.4 mg, 1.2 mol), potassium carbonate (66.2 mg, 0.48 mmol) and ethanol / water (4 mL / 1 mL). The reaction solution was heated to 110 ° C by microwave, and the reaction was stirred for 1 hour. After the reaction was completed, the reaction solution was cooled to room temperature, the mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target pure product E74 (10.6mg, yield: 14.7%, white solid)

¹ H NMR (400 MHz, MeOD): δ 8.07 (s, 1H), 7.85 (brs, 2H), 7.70-7.23 (m, 4H), 3.81 (brs, 1H), 3.25 (s, 2H), 3.15 ( s, 2H), 2.54 (s, 2H), 2.24-1.92 (m, 2H), 1.99 (s, 3H), 1.93 (s, 3H), 1.84 (brs, 2H), 1.58-1.56 (m, 4H) , 1.09 (s, 6H). HPLC: 98.21%@214nm, 98.26%@254nm

LCMS: MS / m / z (ESI): 574.1 [M + H]

Example 75

2-((5-chloro-2-((1- (2- (2-hydroxy-2-methylpropyl) -2-azaspiro [3.5] human-7-yl) -1H-pyrazole- 4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine oxide

Under the protection of argon, sequentially add (2-((2-((1- (2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) amino) to the microwave tube -5-chloropyrimidin-4-yl) amino) phenyl) dimethyl phosphine oxide (150 mg, 0.31 mmol), 2,2-dimethyl ethylene oxide (89.3 mg, 1.24 mol), potassium carbonate (171 mg , 1.24mmol) and ethanol / water (8mL / 2mL). The reaction solution was heated to 110 ° C for 1 hour by microwave. After the reaction was completed, the reaction solution was cooled to room temperature, the mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target pure product E75 (trifluoroacetate, 108mg, yield: 62.8%), properties: white solid

¹ H NMR (400MHz, MeOD): δ 8.35-8.10 (m, 2H), 7.89-6.54 (m, 5H), 4.30-4.28 (m, 1H), 4.21-3.70 (m, 4H), 2.35-2.33 (m, 1H), 2.19-2.17 (m, 1H), 2.15-1.34 (m, 14H), 1.29 (s, 6H).

¹⁹ F NMR (376.5MHz, MeOD): δ-76.91

HPLC: 97.90%@214nm, 98.01%@254nm

LCMS: MS / m (zI): 558.2 [M + H]

Example 76

(2-((5-chloro-2-((1- (2- (2-methoxyethyl) cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl ) Amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine oxide

At room temperature, (2-((5-chloro-2-((1- (cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Group) amino) phenyl) dimethyl phosphine oxide (160mg, 0.30mmol), bromoethylethane (50mg, 0.36mmol) and diisopropylethylamine (116mg, 0.90mmol) dissolved in N, N-di Methyl formamide (1mL). The reaction solution was heated to 70 ° C and stirred for 12 hours. LCMS detected the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the residue was subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure product E76 (7mg, yield: 5%), properties: white solid

¹ H NMR (400MHz, DMSO) δ 9.26 (s, 1H), 8.13 (s, 1H), 7.65-7.53 (m, 3H), 7.37 (brs, 1H), 7.22 (brs, 1H), 4.69 (brs , 1H), 3.46-3.43 (m, 2H), 3.26 (s, 3H), 2.64 (brs, 2H), 2.54 (br, 2H), 2.50-2.38 (m, 4H), 2.02 (brs, 2H), 1.84-1.75 (m, 2H), 1.78 (s, 3H), 1.75 (s, 3H).

HPLC: 98.55%@214nm, 99.82%@254nm

LCMS: Rt: 1.00 min; MS m / z (ESI): 530.5 [M + H].

Example 77

(2-((5-chloro-2-((1- (2- (2-methoxyethyl) -2-azaspiro [3.5] human-7-yl) -1H-pyrazole-4- Yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylsulfide Phosphine

At room temperature, (2-((2-((1- (2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) directly) -5-chloropyrimidine-4- Yl) amino) phenyl) dimethylphosphine sulfide (60mg, 0.12mmol), 1-bromo-2-methoxyethane (33.1mg, 0.24mol) and diisopropylethylamine (29.3mg, 0.24 mmol) was dissolved in acetonitrile (5 mL), and then the reaction solution was heated to 80 ° C, and the reaction was stirred for 16 hours. After the reaction is complete, the reaction solution is cooled to room temperature and filtered, the filtrate is concentrated under reduced pressure, and the residue is prepared by high-performance liquid chromatography (eluent gradient:

) Purification, to obtain the target pure product E77 (trifluoroacetate, 8.7mg, yield: 13%), properties: white solid

¹ H NMR (400 MHz, MeOD): δ 8.07 (s, 1H), 7.86 (brs, 2H), 7.74-6.99 (m, 4H), 3.81 (brs, 1H), 3.45-3.42 (m, 2H), 3.34 (s, 3H), 3.20 (s, 2H), 3.09 (s, 2H), 2.72 (t, J = 4.0Hz, 2H), 2.15-1.92 (m, 2H), 2.02 (s, 3H), 1.99 (s, 3H), 1.87-1.85 (m, 2H), 1.59-1.55 (m, 4H).

¹⁹ F NMR (376.5MHz, MeOD): δ-76.94

HPLC: 94.66%@214nm, 94.71%@254nm

LCMS: MS / m (zI): 560.2 [M + H]

Example 78

(2-((5-chloro-2-((5-chloro-1- (2- (2-methoxyethyl) -2-azaspiro [3.5] nonane-7-yl) -1H- Pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) di Methylphosphine oxide

7- (5-chloro-4-amino-1H-pyrazol-1-yl) -2- (2-methoxyethyl) -2-azaspiro [3.5] nonane (70mg, 0.23mmol) , (2-((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (88mg, 0.28mmol) and ammonium chloride (36mg, 0.69mmol) were dissolved in ethanol (3mL ). The reaction solution was stirred at 100 ° C for 8 hours. The reaction was completed by LCMS. The reaction solution was cooled to room temperature and filtered. The filtrate was prepared by preparative high-performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E78 (trifluoroacetate salt, 9.4 mg, yield: 7.1%), character: white solid.

¹ H NMR (400 MHz, MeOD) δ: 8.41-8.05 (m, 2H), 7.68-7.54 (m, 3H), 7.35-7.31 (m, 1H), 4.36-4.34 (m, 1H), 4.21-4.18 ( m, 1H), 4.04-3.90 (m, 3H), 3.64-3.60 (m, 2H), 3.47-3.40 (m, 2H), 3.40 (s, 3H), 2.32-2.17 (m, 2H), 1.98- 1.78 (m, 12H).

¹⁹ F NMR (376.5MHz, MeOD) δ: -77.30

HPLC: 97.723%@214nm, 98.827%@254nm

LCMS: MS / m (zI): 578.4 [M + H]

Example 79

(2-((5-chloro-2-((1- (1,2,2,2-trifluoroethyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine- 4-yl) amino) phenyl) dimethylphosphine oxide Thing

At room temperature, (2-((5-chloro-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine oxide (100 mg, 0.22 mmol), 2,2,2-trifluoroethyl trifluoromethanesulfonate (102 mg, 0.44 mmol) and diisopropylethylamine (142 mg, 1.1 mmol) were dissolved In acetonitrile (4mL), the reaction was stirred at 70 ° C for 12 hours; after the reaction was complete, the reaction solution was cooled and concentrated under reduced pressure, and the residue was subjected to high performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E79 (18.5 mg, yield: 16%), character: white solid.

¹ H NMR (400MHz, DMSO-d6) δ: 11.25-10.52 (m, 1H), 9.28 (s, 1H), 8.14 (s, 1H), 7.64-7.55 (m, 3H), 7.41 (brs, 1H) , 7.22 (brs, 1H), 4.01 (brs, 2H), 3.28-3.22 (m, 2H), 3.01-2.98 (m, 2H), 2.53 (m, 1H), 1.88 (br, 4H), 1.78 (s , 3H), 1.75 (s, 3H).

¹⁹ F NMR (376.5MHz, MeOD) δ: -68.18

HPLC: 97.62%@214nm, 98.64%@254nm

LCMS: MS / m (zI): 528.4 [M + H]

Example 80

(2-((5-chloro-2-((1- (1- (2,2-difluoroethyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethylphosphine oxide

At room temperature, (2-((5-chloro-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine oxide (100 mg, 0.22 mmol), 1,1-difluoro-2-iodoethane (84 mg, 0.44 mmol) and potassium carbonate (121 mg, 0.88 mmol) were added to N, N-dimethylformamide (4mL), the reaction was stirred at 80 ° C for 12 hours; after the reaction was complete, the reaction liquid was cooled and concentrated under reduced pressure, and the residue was prepared by high-performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E80 (40 mg, yield: 36%), character: white solid.

¹ H NMR (400MHz, DMSO-d6) δ: 11.50-10.50 (m, 1H), 9.27 (s, 1H), 8.14 (s, 1H), 7.75-7.65 (m, 2H), 7.58-7.54 (m, 1H), 7.41 (br, 1H), 7.23 (br, 1H), 6.17 (t, J = 56.0 Hz, 1H), 3.99 (br, 1H), 3.00-2.97 (m, 2H), 2.80-2.73 (m , 2H), 2.35-2.32 (m, 2H), 1.88 (br, 4H), 1.78 (s, 3H), 1.75 (s, 3H).

¹⁹ F NMR (376.5MHz, DMSO-d6) δ: -118.83

HPLC: 97.49%@214nm, 97.87%@254nm

LCMS: MS / m (zI): 510.4 [M + H]

Example 81

2-((5-chloro-2-((1- (2- (dimethylamino) ethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) di Methylphosphine oxide

At room temperature, N, N-dimethyl-2- (4-amino-1H-pyrazol-1-yl) ethylamine (100 mg, 0.65 mmol), (2-((2,5-dichloropyrimidine- 4-yl) amino) phenyl) dimethylphosphine oxide (205 mg, 0.65 mmol), trifluoroacetic acid (661 mg, 6.5 mmol), dissolved in ethanol (5 mL), and the reaction solution was stirred at 50 ° C for 12 hours; After the reaction is complete, the reaction liquid is cooled and concentrated under reduced pressure, and the residue is subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E81 (trifluoroacetate, 93 mg, yield: 33%), properties: yellow solid.

¹ H NMR (400MHz, DMSO-d6): δ 11.19-10.79 (m, 1H), 9.31 (s, 1H), 8.76 (br, 0.4H), 8.14 (s, 1H), 7.83-7.40 (m, 4H), 7.21 (brs, 1H), 4.19 (brs, 2H), 2.88 (brs, 2H), 2.35 (brs, 6H), 1.79 (s, 3H), 1.76 (s, 3H).

¹⁹ F NMR (376.5MHz, DMSO-d6) δ: -73.42

HPLC: 99.68%@214nm, 99.80%@254nm

LCMS: MS / m (zI): 434.4 [M + H]

Example 82

(2-((5-chloro-2-((1- (1- (2-hydroxy-2-methylpropyl) piperidin-4-yl) -5-methyl-1H-pyrazole-4- Yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine sulfide

At room temperature, convert (2-((5-chloro-2-((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) (Amino) phenyl) dimethyl phosphine sulfide (81mg, 0.17mmol), 2,2-dimethyl ethylene oxide (125mg, 1.7mmol) and potassium carbonate (120mg, 0.87mmol) dissolved in ethanol / water (2 mL / 0.5mL). The reaction solution was heated to 110 ° C in a microwave reactor, and the reaction was stirred for 20 minutes. LCMS detected the reaction was complete, and the reaction mixture was concentrated under reduced pressure. The residue is subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure product E82 (6mg, yield: 6%), character: white solid.

¹ H NMR (400MHz, DMSO) δ 9.38 (s, 1H), 8.57 (br, 1H), 8.11 (s, 1H), 7.93 (br, 1H), 7.74-7.63 (m, 1H), 7.51 (brs , 1H), 7.35-7.30 (m, 2H), 4.07 (s, 1H), 3.94 (brs, 1H), 3.04-3.01 (m, 2H), 2.28-2.18 (m, 4H), 2.07-1.92 (m , 11H), 1.67-1.60 (m, 2H), 1.10 (s, 6H).

HPLC: 97.59% (214nm), 97.15% (254nm)

LCMS: Rt: 1.16 min; MS m / z (ESI): 548.4 [M + H].

Example 83

(2-((5-chloro-2-((1- (2-hydroxy-2-methylpropyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Radical) amino) phenyl) dimethyl phosphine oxidation Thing

At room temperature, (2-((5-chloro-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine oxide (100mg, 0.22mmol), 2,2-dimethylethylene oxide (162mg, 2.2mmol), potassium carbonate (155mg, 1.1mmol) dissolved in ethanol (3mL) and water (0.75mL) ) In the mixed solution, the reaction was heated to 110 ° C. with microwave and the reaction was stirred for 20 minutes. After the reaction is complete, the reaction solution is cooled and concentrated under reduced pressure, and the residue is subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain pure target product E83 (58 mg, yield: 50%), character: white solid.

¹ H NMR (400MHz, DMSO-d6) δ: 11.35-10.50 (m, 1H), 9.28 (s, 1H), 8.76 (br, 1H), 8.14 (s, 1H), 8.10-7.57 (m, 3H) , 7.39 (brs, 1H), 7.22 (brs, 1H), 4.10 (s, 1H), 3.94 (brs, 1H), 3.04-3.01 (m, 2H), 2.23 (br, 4H), 1.84 (br, 4H ), 1.78 (s, 3H), 1.75 (s, 3H), 1.12 (s, 6H).

HPLC: 99.66%@214nm, 99.85%@254nm

LCMS: MS / m (zI): 518.4 [M + H]

Example 84

(2-((5-chloro-2-((1- (1- (2- (dimethylamino) ethyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine -4-yl) amino) phenyl) dimethylphosphine oxide Thing

At room temperature, (2-((5-chloro-2-((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine oxide (100 mg, 0.22 mmol), 2-chloro-N, N-dimethylethylamine hydrochloride (32 mg, 0.22 mmol), sodium carbonate (70 mg, 0.66 mmol) and sodium iodide (7 mg , 0.044mmol) was added to N'N-dimethylformamide (4mL), the reaction was stirred at 80 ℃ for 12 hours; after the reaction was complete, the reaction solution was cooled and concentrated under reduced pressure, the residue was subjected to high performance liquid chromatography (Eluent gradient:

) Purification to obtain the pure target product E84 (58 mg, yield: 50%), character: white solid.

¹ H NMR (400MHz, DMSO-d6) δ: 11.16-10.68 (m, 1H), 9.27 (s, 1H), 8.77 (br, 0.3H), 8.13 (s, 1H), 7.63-7.54 (m, 3H) ), 7.40 (br, 1H), 7.22 (br, 1H), 3.97 (br, 1H), 2.96-2.93 (m, 2H), 2.41-2.35 (m, 4H), 2.15 (s, 6H), 2.09- 2.03 (m, 2H), 1.88 (br, 4H), 1.78 (s, 3H), 1.75 (s, 3H).

HPLC: 97.85%@214nm, 98.65%@254nm

LCMS: MS / m (zI): 517.6 [M + H]

Example 85

2-((5-chloro-2-((1- (2-methyl-cyclopenta [c] pyrrole-5-acyl) -1h-pyrazol-4-acyl) amino) pyrimidine-4-acyl) Amino) phenyl) dimethylphosphine sulfide

At room temperature, (2-((5-chloro-2-((1- (cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Base) amino) phenyl) dimethyl phosphine sulfide (80 mg, 0.16 mmol) was dissolved in methanol (4 mL), triethylamine was added dropwise to make the solution alkaline, and then 30% aqueous formaldehyde solution (300 mg, 3.2 mmol) was added, The reaction was stirred at room temperature for 12 hours; additional sodium triacetate borohydride (339 mg, 1.6 mmol) was added and stirred at room temperature for 1 hour; after the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was subjected to high-performance liquid chromatography (washing) Removal gradient:

) Purification to obtain the pure target product E85 (34 mg, yield: 42%), character: white solid.

¹ H NMR (400MHz, DMSO-d6) δ: 9.32-9.26 (m, 2H), 8.15 (brs, 1H), 7.99 (brs, 1H), 7.69 (brs, 2H), 7.49 (brs, 1H), 7.18 (brs, 2H), 4.46 (brs, 1H), 2.63 (brs, 2H), 2.37 (brs, 4H), 2.25 (s, 3H), 1.98 (s, 3H), 1.85 (s, 3H), 1.85 ( brs, 2H), 1.73 (brs, 2H).

HPLC: 97.79%@214nm, 98.11%@254nm

LCMS: MS / m (zI): 502.4 [M + H]

Example 86

(2-((5-Bromo-2-((1-methylcyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino ) Phenyl) dimethyl phosphine oxide

At room temperature, 2-methyl-5- (4-amino-1H-pyrazol-1-yl) cyclopentyl [c] pyrrolidine (62mg, 0.3mmol), (2-((5-bromo- 2-chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (108 mg, 0.3 mmol) and ammonium chloride (160 mg, 3.0 mmol) were dissolved in isopropanol (3 mL). The reaction solution was stirred at 80 ° C for 12 hours. After LCMS detected the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the residue was subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure product E86 (15mg, yield: 10%), character: white solid.

¹ H NMR (400MHz, DMSO) δ 10.90-10.31 (m, 1H), 9.28 (brs, 1H), 8.20 (s, 1H), 8.02 (brs, 1H), 7.65-7.54 (m, 3H), 7.30 -7.25 (m, 2H), 4.61 (brs, 1H), 2.65 (brs, 2H), 2.41-2.34 (m, 4H), 2.23 (s, 3H), 1.99 (brs, 2H), 1.77-1.73 (m , 2H), 1.76 (s, 3H), 1.73 (s, 3H).

HPLC: 94.28%@214nm, 94.22%@254nm

LCMS: Rt: 0.93 min; MS m / z (ESI): 532.4 [M + H].

Example 87

(2-((5-trifluoromethyl-2-((1- (2- (2,2,2-trifluoroethyl) cyclopenta [c] pyrrolidin-5-yl) -1H- Pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethyl phosphine oxide

At room temperature, (2-((2-chloro-5- (trifluoromethyl) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (119 mg, 0.34 mmol), (2-(( 5-chloro-2-((1- (cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine oxide (70 mg, 0.34 mmol) and ammonium chloride (180 mg, 3.4 mmol) were dissolved in ethanol (5 mL), the reaction system was heated to reflux, and the reaction was stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the target pure product E87 (19.6mg, yield: 11.1%, white solid)

¹ H NMR (400 MHz, MeOD) δ: 8.30-8.26 (m, 1H), 7.91 (brs, 1H), 7.73-7.55 (m, 3H), 7.27 (brs, 1H), 7.21 (brs, 1H), 4.21 (brs, 1H), 2.79 (brs, 4H), 2.37 (brs, 3H), 2.25 (brs, 2H), 1.95-1.74 (m, 10H).

¹⁹ F NMR (400MHz, MeOD) δ: -62.90

HPLC: 98.165%@214nm, 99.338%@254nm

LCMS: MS / m (zI): 520.1 [M + H]

Example 88

(2-((5-chloro-2-((1- (3- (4- (2-methoxyethyl) piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl ) Amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine Oxide

(2-((5-chloro-2-((1- (3- (piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) (Amino) phenyl) dimethylphosphine oxide (40mg, 0.079mmol), bromoethyl methyl ether (21mg, 0.158mmol) and N, N-isopropylethylamine (30mg, 0.237mmol) dissolved in N, N- Dimethylformamide (3mL). The reaction solution was stirred at room temperature for 10 hours. The reaction was detected by LCMS. The reaction solution was directly prepared by high-performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E88 (8 mg, yield: 18.2%), character: white solid.

¹ H NMR (400 MHz, MeOD) δ: 8.41-8.05 (m, 1H), 8.05 (s, 1H), 7.68-7.59 (m, 3H), 7.49 (br, 1H), 7.33 (br, 1H), 4.71 (brs, 1H), 3.55 (t, J = 5.6 Hz, 2H), 3.34 (s, 3H), 3.07-3.04 (m, 1H), 2.63-2.47 (m, 14H), 1.85 (s, 3H), 1.81 (s, 3H).

HPLC: 82.874%@214nm, 99.553%@254nm

LCMS: MS / m (zI): 559.3 [M + H]

Example 89 and Example 90

(2-((5-chloro-2-((1- (1- (2-methoxyethyl) piperidin-3-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Radical) amino) phenyl) dimethyl phosphine oxide (Example 89, single unknown configuration isomer, Rt: 3.313min; Example 90, single unknown configuration isomer, Rt: 3.949min)

At room temperature, tert-butyl 3- (4-((5-chloro-4-((2- (dimethylphosphinooxy) phenyl) amino) pyrimidin-2-yl) amino) -1H-pyrazole -1-yl) piperidine-1-carboxylate (400 mg, 0.73 mmol) was dissolved in a solution of hydrogen chloride in dioxane (4M, 5 mL) and methanol (0.5 mL), and the reaction mixture was reacted at room temperature for 0.5 hour. LCMS detected the reaction was complete, the reaction solution was concentrated under reduced pressure, and then dissolved with N, N-dimethylformamide (5mL), and then added bromoethyl methyl ether (100mg, 0.73mmol) and N, N'-diisopropyl Ethyl amine (283mg, 2.19mmol), after the reaction mixture was reacted at room temperature for 10 hours, the reaction was completed by LCMS, and the reaction solution was subjected to high performance liquid chromatography (eluent gradient:

) Purification to obtain pure racemic product (150mg, yield: 40.8%) Properties: yellow solid, chiral separation of target product:

Chiral separation of the racemic product (150mg, 0.3mmol) (separation conditions:

Instrument signal: Waters-SFC80

Column model: IA-H (2.5 × 25cm, 10μm)

Mobile phase A: Supercritical carbon dioxide,

Mobile phase B: ethanol / ammonia

A: B = 70/30, flow rate: 70g / min

Detection wavelength: 214nm

Column temperature: 25 ℃

Column pressure: 100bar)

The target product E89 (single unknown configuration isomer, Rt: 3.313min) (54.93mg, yield: 36.6%, traits: yellow solid) and target product E90 (single unknown configuration isomer, Rt: 3.949min) were obtained ) (59.64 mg, yield: 39.766%, character: yellow solid).

E89

¹ H NMR (400MHz, MeOD) δ: 8.41-8.05 (m, 2H), 7.70-7.62 (m, 3H), 7.46 (br, 1H), 7.35 (br, 1H), 4.24 (br, 1H), 3.60 -3.57 (m, 2H), 3.35 (s, 3H), 3.31-3.28 (m, 3H), 3.06-3.03 (m, 1H), 2.81 (br, 2H), 2.03 (br, 1H), 1.85 (s , 3H), 1.81 (s, 3H), 1.85-1.75 (m, 3H).

Chiral HPLC: ee 100%, 254nm, Rt: 3.313min

HPLC: 99.701%@214nm, 99.548%@254nm

LCMS: MS / m (zI): 504.3 [M + H]

E90

¹ H NMR (400MHz, MeOD) δ: 8.41-8.05 (m, 2H), 7.70-7.61 (m, 3H), 7.46 (s, 1H), 7.35 (br, 1H), 4.24 (s, 1H), 3.60 -3.57 (m, 2H), 3.35 (s, 3H), 3.31-3.30 (m, 3H), 3.04-3.02 (m, 1H), 2.81 (br, 2H), 2.03 (br, 1H), 1.85 (s , 3H), 1.81 (s, 3H), 1.85-1.75 (m, 3H).

Chiral HPLC: ee 99.16%, 254nm, Rt: 3.949min

HPLC: 97.808%@214nm, 99.787%@254nm

LCMS: MS / m (zI): 504.3 [M + H]

Example 91 and Example 92

(2-((5-chloro-2-((1- (1- (2-hydroxy-2-methylpropyl) piperidin-3-yl) -1H-pyrazol-4-yl) amino) pyrimidine -4-yl) amino) phenyl) dimethylphosphine oxide (Example 91, unknown single isomer 1, Rt: 2.784 min; Example 92, unknown single isomer 2, Rt: 4.559 min)

At room temperature, (2-((5-chloro-2-((1- (piperidin-3-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine oxide (310mg, 0.7mmol), 2,2-dimethylethylene oxide (62mg, 0.84mmol), potassium carbonate (193mg, 1.4mmol) dissolved in ethanol / water (4/1, 10mL) ) In the mixed solvent, the system was heated to 110 ° C for 3 hours with microwave. After the reaction was detected by LCMS, the reaction solution was filtered, and the filtrate was directly subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the racemic target product (160mg, yield: 20%), character: white solid

LCMS: Rt: 1.168 min; MS m / z (ESI): 518 [M + H].

The racemic target product (160mg, 0.3mmol) was separated by supercritical chiral high performance liquid phase (separation conditions:

Instrument signal: Waters-SFC80

Column model: IA-H (2.5 * 25cm, 10μm)

Mobile phase A: Supercritical carbon dioxide,

Mobile phase B: ethanol / ammonia

A: B = 60/40, flow rate: 70g / min

Detection wavelength: 214nm

Column temperature: 25 ℃

Column pressure: 100 bar), two single isomers are obtained:

E91 (unknown single isomer 1, Rt: 2.784min) (10mg, yield: 6%), properties: off-white solid

¹ H NMR (400MHz, MeOD) δ 8.20 (br, 1H), 8.06 (s, 1H), 7.68-7.63 (m, 3H), 7.44-7.35 (m, 2H), 4.21 (brs, 1H), 3.13 (br, 1H), 2.93 (br, 1H), 2.55-2.30 (m, 4H), 1.97 (br, 1H), 1.85-1.75 (m, 10H), 1.21 (s, 6H).

Chiral HPLC: ee: 100%, Rt: 2.784min

HPLC: 98.051%@214nm; 99.530%@254nm

LCMS: MS / m (zI): 518.4 [M + H]

E92 (unknown single isomer 2, Rt: 4.559min) (50mg, yield: 30%), properties: off-white solid

¹ H NMR (400MHz, MeOD) δ 8.20 (br, 1H), 8.06 (s, 1H), 7.68-7.62 (m, 3H), 7.44-7.35 (m, 2H), 4.21 (brs, 1H), 3.13 (br, 1H), 2.96 (br, 1H), 2.55-2.30 (m, 4H), 1.97 (br, 1H), 1.85-1.75 (m, 10H), 1.21 (s, 6H).

Chiral HPLC: ee: 100%, Rt: 4.559min

HPLC: 97.927%@214nm; 99.808%@254nm

LCMS: MS / m (zI): 518.4 [M + H]

Example 93

(2-((5-chloro-2-((1- (3- (2-hydroxy-2-methylpropyl) -3-azaspiro [5.5] undecane-9-yl) -1H- Pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethyl phosphine oxide

At room temperature, (2-((2-((1- (3-azaspiro [5.5] undec-9-yl) -1H-pyrazol-4-yl) amino) -5-chloropyrimidine- 4-yl) amino) phenyl) dimethylphosphine oxide (30 mg, 0.05 mmol), 2,2-dimethylethylene oxide (36 mg, 0.5 mmol) and potassium carbonate (42 mg, 0.25 mmol) dissolved in ethanol / Water (2mL / 0.5mL). The reaction solution was heated to 110 ° C in a microwave reactor and stirred for 20 minutes. LCMS detected the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the residue was then subjected to high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure product E93 (hydrochloride, 6mg, yield: 20%), properties: white solid;

¹ H NMR (400MHz, MeOD) δ 8.25 (brs, 1H), 8.05 (s, 1H), 7.70-7.60 (m, 3H), 7.43 (s, 1H), 7.34 (brs, 1H), 3.92 (brs , 1H), 2.74-2.71 (m, 4H), 2.48 (s, 2H), 1.89-1.82 (m, 12H), 1.72 (brs, 2H), 1.54-1.46 (m, 2H), 1.35-1.27 (m , 2H), 1.21 (s, 6H).

HPLC: 98.99%@214nm, 99.38%@254nm

LCMS: Rt: 0.98 min; MS m / z (ESI): 586.3 [M + H].

Example 94

(2-((5-chloro-2-((1- (3- (4- (2-hydroxy-2-methylpropyl) piperazin-1-yl) cyclobutyl) -1H-pyrazole- 4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine sulfide

At room temperature, (2-((5-chloro-2-((1- (3- (piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethyl phosphine sulfide (33mg, 0.064mmol), 2,2-dimethyl ethylene oxide (46mg, 0.64mmol), potassium carbonate (45mg, 0.32mmol) dissolved in ethanol ( 2mL) and water (0.5mL), the reaction was heated to 110 ° C with a microwave and stirred for 20 minutes; after the reaction was completed, the reaction liquid was cooled to room temperature, then concentrated under reduced pressure, and the residue was prepared by high-performance liquid chromatography ( Eluent gradient:

) Purification to obtain the pure target product E94 (2mg, yield: 5%), properties: white solid.

¹ H NMR (400 MHz, MeOD–d4) δ: 8.08 (s, 1H), 7.85 (brs, 2H), 7.65 (t, J = 8.0 Hz, 1H), 7.46 (br, 1H), 7.42 (s, 2H ), 4.61 (brs, 1H), 3.03 (brs, 1H), 2.69 (brs, 4H), 2.45 (br, 8H), 2.34 (s, 2H), 2.02 (s, 3H), 1.98 (s, 3H) , 1.19 (s, 6H).

HPLC: 98.11%@214nm, 98.74%@254nm

LCMS: MS / m (zI): 589.1 [M + H]

Example 95

2-((5-chloro-2-((1- (3- (4-ethylpiperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl ) Amino) phenyl) dimethylphosphine sulfide

At room temperature, (2-((5-chloro-2-((1- (3- (piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethylphosphine sulfide (33mg, 0.064mmol) was dissolved in ethanol (3mL), triethylamine was added dropwise to make the solution alkaline, and then 1M acetaldehyde tetrahydrofuran solution (1.3mL, 1.28 mmol), the reaction was stirred at room temperature for 12 hours; additional sodium triacetate boron hydride (136 mg, 0.64 mmol) was added, and the reaction was further stirred at room temperature for 1 hour; after the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was separated by a high-performance liquid phase Preparative chromatography (eluent gradient:

) Purification to obtain the pure target product E95 (trifluoroacetate salt, 4 mg, yield: 11%), properties: white solid.

¹ H NMR (400 MHz, MeOD-d4): δ 8.14 (s, 1H), 7.92 (br, 1H), 7.82 (br, 1H), 7.70-7.67 (m, 1H), 7.54-7.42 (m, 3H ), 4.62 (brs, 1H), 3.32 (br, 4H), 3.31-3.25 (m, 4H), 3.20-2.60 (m, 3H), 2.53 (br, 4H), 2.04 (s, 3H), 2.00 ( s, 3H), 1.36 (t, J = 8.0Hz, 3H).

¹⁹ F NMR (376.5MHz, MeOD-d4): δ-77.25

HPLC: 96.29%@214nm, 97.86%@254nm

LCMS: MS / m (zI): 545.4 [M + H]

Example 96

(2-((5-chloro-2-((1- (3- (4- (2-methoxyethyl) piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl ) Amino) pyrimidin-4-yl) amino) phenyl) dimethylsulfide Phosphine

At room temperature, (2-((5-chloro-2-((1- (3- (piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidine-4 -Yl) amino) phenyl) dimethylphosphine sulfide (33mg, 0.064mmol), bromoethyl methyl ether (11mg, 0.077mmol), diisopropylethylamine (41mg, 0.32mmol) dissolved in N'N -In dimethylformamide (3mL), the reaction was stirred at 50 ° C for 12 hours; after the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was subjected to high performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E96 (2mg, yield: 5%), character: white solid.

¹ H NMR (400 MHz, MeOD–d4) δ: 8.08 (s, 1H), 7.87 (brs, 2H), 7.67-7.63 (m, 1H), 7.42 (br, 3H), 4.59 (br, 1H), 3.55 (t, J = 5.6 Hz, 2H), 3.34 (s, 3H), 3.03 (br, 1H), 2.62 (t, J = 5.6 Hz, 2H), 2.62 (br, 2H), 2.45 (brs, 6H) , 2.02 (s, 3H), 1.99 (s, 3H), 1.33-1.29 (m, 4H).

HPLC: 88.62%@214nm, 89.04%@254nm

LCMS: MS / m (zI): 575.5 [M + H]

Example 97

2-((5-chloro-2-((1- (1-methylpiperidin-3-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) di Methylphosphine oxide

At room temperature, (2-((5-chloro-2-((1- (piperidin-3-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) Dimethylphosphine oxide (320mg, 0.72mmol) was dissolved in methanol (4mL), triethylamine was added dropwise to make the solution alkaline, then 30% aqueous formaldehyde solution (1.3g, 14.4mmol) was added, and the reaction was stirred at room temperature 12 hours; additional sodium triacetylborohydride (916 mg, 4.32 mmol) was added, and stirring was continued at room temperature for 1 hour. After the reaction was completed, the reaction was concentrated under reduced pressure, and the residue was subjected to high-performance liquid chromatography (eluent gradient:

) Purification to obtain the pure target product E97 (110 mg, yield: 34%), character: white solid.

¹ H NMR (400 MHz, DMSO-d6): δ 11.30-10.65 (m, 1H), 9.26 (s, 1H), 8.14 (m, 2H), 7.62 (br, 1H), 7.54 (t, J = 8.0 Hz, 1H), 7.42 (br, 1H), 7.21 (br, 1H), 4.11 (br 1H), 2.92-2.90 (m, 1H), 2.70-2.68 (m, 1H), 2.19 (s, 3H), 2.10 (br, 1H), 1.93-1.90 (m, 2H), 1.79 (s, 3H), 1.75 (s, 3H), 1.70 (br, 1H), 1.59 (br, 2H).

HPLC: 98.94%@214nm, 99.41%@254nm

LCMS: MS / m (zI): 460.3 [M + H]

Example 98

(2-((5-chloro-2-((1- (3- (4-ethylpiperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Radical) amino) phenyl) dimethyl phosphine oxide

Add (2-((5-chloro-2-((1- (3- (piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) to a 100mL single-necked bottle Pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (80mg, 0.16mmol) and dissolved in ethanol (5mL), then added acetaldehyde in tetrahydrofuran solution (0.64mL, 0.64mmol), the compound was stirred for 10 hours After adding sodium triacetoxyborohydride (67mg, 0.32mmol), the reaction mixture continued to react at room temperature for 1 hour, and the reaction was completed by LCMS. The reaction solution was diluted with acetonitrile (2mL), and then used high performance liquid preparative chromatography (eluent gradient:

) Purification to obtain pure product E98 (4mg, yield: 4.7%), character: white solid.

¹ H NMR (400MHz, MeOD): δ8.41-8.05 (m, 1H), 8.05 (s, 1H), 7.71-7.60 (m, 3H), 7.49 (br, 1H), 7.34 (br, 1H), 4.70-4.68 (m, 1H), 3.09-3.05 (m, 1H), 2.65-2.45 (m, 14H), 1..85 (s, 3H), 1.82 (s, 3H), 1.12 (t, J = 7.2Hz, 3H).

HPLC: 95.261%@214nm, 95.304%@254nm

LCMS: MS / m (zI): 529.4 [M + H]

Example 99

(2-((5-chloro-2-((1- (2- (2-oxetan-3-yl) cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazole- 4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl Phosphine oxide

(2-((5-chloro-2-((1- (cyclopenta [c] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino ) Phenyl) dimethylphosphine oxide (100mg, 0.212mmol) was dissolved in methanol (5mL), and then oxetan-3-one (76mg, 1.06mmol) and sodium cyanoborohydride (65mg, 1.06mmol) were added The reaction mixture was reacted at room temperature for 10 hours. LCMS detected the reaction was complete. The reaction solution was prepared by high-performance liquid chromatography (eluent gradient:

) Purification to obtain pure product E99 (22 mg, yield: 20.4%), character: white solid.

¹ H NMR (400MHz, MeOD) δ: 8.41-8.05 (m, 2H), 7.69-7.60 (m, 3H), 7.40-7.35 (m, 2H), 4.75-4.71 (m, 3H), 4.65-4.61 ( m, 2H), 3.58-3.54 (m, 1H), 2.79 (brs, 2H), 2.65-2.60 (m, 2H), 2.33-2.29 (m, 2H), 2.15-2.03 (m, 2H), 1.96 1.93 (m, 2H), 1.84 (s, 3H), 1.81 (s, 3H).

HPLC: 98.816%@214nm, 98.867%@254nm

LCMS: MS / m (zI): 528.4 [M + H]

LCMS: MS m / z (ESI): 463.1 [M + H].

Test Example 1: Enzymatic EGFR phosphorylation inhibition experiment

Experimental Materials:

HTRF KinEASE-TK was purchased from CisBio (France).

The 384-well assay plate was purchased from greater bio-one (Germany).

The 384-well source plate was purchased from LABCYTE (USA).

MgCl ₂ , MnCl ₂ , DTT, TritonX-100, HEPES, BSA were purchased from Sigma (USA).

EGFRWT-WT, EGFRWT-del19 / T790M / C797S, EGFRWT-L858R / T790M / C797S, EGFRWT-del19 / T790M, EGFRWT-L858R / T790M are provided by Jiangsu Xiansheng Pharmaceutical Co., Ltd.

laboratory apparatus:

Automatic micropore pipette: Precision PRC384U (BioTek, USA)

Nano upgrade sound wave shift system:

HANDLE RS (LABCYTE, USA)

Multi-label detection analyzer: Envision Multilabel Reader (PerkinElmer, USA)

experimental method:

1. Prepare a kinase buffer solution (20 mM HEPES pH 7.5, 0.01% Triton X-100, 0.01% BSA, 1 mM DTT, 1 mM MnCl ₂ , 5 mM MgCl ₂ ).

2. Preparation of the compound. The final detection concentration of the compound is 10 μM, which is formulated into a 100-fold concentration, ie 1 mM. Use an automatic micro-pipette (Precision PRC384U) to make a 4-fold gradient dilution of the test compound to obtain 10 dose concentrations ranging from 1mM to 3.8nM. Use Echo to transfer the test compound to a 384-well plate at 100nL per well Compound.

3. Kinase reaction. The enzyme reaction solution was prepared using kinase buffer, 5 μl of the enzyme reaction solution was transferred to the reaction well of the 384-well plate, and 5 μl of kinase buffer was added to the negative control well. Shake well at 450 rpm and incubate at room temperature for 10 minutes. Use kinase buffer to prepare the substrate solution, transfer 5 μl of the substrate solution to the reaction well of the 384-well plate to start the reaction, shake and mix at 450 rpm, and incubate at room temperature for 60 minutes.

4. Terminate the reaction. Add 10 μl of reaction stop solution (Streptaridin-XL665, TK Antibody-Cryptate) to the 384-well reaction plate, centrifuge at 1000 rpm for 1 minute, leave at room temperature for 60 minutes, and read the plate with Envision

5. Curve fitting. Copy the data of the luminescence reading from the Envision program, and convert the luminescence reading ratio of 665nM / 615nM to the percentage of inhibition by the formula: Percent inhibition = (max-signal ratio) / (max-min) * 100. "Min" is the ratio of the fluorescence reading of the control sample without enzyme; "max" is the ratio of the fluorescence reading of the sample added DMSO as a control. Import the data into MS Excel and use XLFit excel add-in version 5.4.0.8 for curve fitting: Y = Bottom + (TopBottom) / (1+ (IC ₅₀ / X) ^ HillSlope).

The enzymatic activity of BaF ₃ (WT-WT) of the compound of the present invention inhibits IC ₅₀ , the enzymatic activity of BaF3 (WT-del19 / T790M / C797S) inhibits the enzyme of IC ₅₀ , BaF ₃ (WT-L858R / T790M / C797S) Science inhibitory activity _{IC 50, BaF 3 (WT-} del19 / T790M) of enzymatic activity inhibition _{IC 50, BaF 3 (WT-} L858R / T790M) inhibition of enzymatic activity below ₅₀ IC.

It can be seen from the following results that most of the compounds of the present invention have a good inhibitory effect on four mutant enzymes, and the anti-proliferative activity is below 100 nM. Most of the compounds of the present invention have very good selectivity for wild-type EGFR, for example, compounds E1 ′, E26, and E30 have an inhibitory activity on BaF3 (WT-del19 / T790M / C797S), and IC ₅₀ inhibits BaF3 (WT- The enzymatic activity of WT) inhibits IC ₅₀ by more than 150 times, which is significantly better than that of Brigatinib.

Test Example 2 Cell anti-proliferation experiment

Experimental Materials:

RPMI1640 was purchased from Gibco (USA).

FBS was purchased from Hyclone (USA).

IL-3 was purchased from R & D (USA).

DMSO was purchased from Amresco (USA).

Luminescent Cell Viability kit was purchased from Promega (USA).

BaF3 cells were purchased from Riken (Japan).

BaF3 / EGFR-WT, BaF3 / EGFR-Del19 / T790M / C797S and BaF3 / EGFR-L858R / T790M / C797S cells were constructed by Sino-American Crown Biotech (Taicang) Co., Ltd.

BaF3 medium: RPMI1640 + 10% FBS + 8ng / ml IL-3.

BaF3 / EGFR-WT, BaF3 / EGFR-Del19 / T790M / C797S and BaF3 / EGFR-L858R / T790M / C797S medium: RPMI1640 + 10% FBS.

Plate reader: EnVision (PerkinElmer, USA).

experimental method:

Adjust the cell concentration, each kind of 2000 BaF ₃ , BaF ₃ / EGFR-WT, BaF ₃ / EGFR-Del19 / T790M / C797S or BaF ₃ / EGFR-L858R / T790M / C797S cells in a 96-well plate, 90 μl volume, use DMSO Prepare a 1000X test compound mother liquor, and use DMSO to make a 3-fold gradient of the test compound (a total of 9 concentrations), and then make a 100-fold dilution with the corresponding cell culture medium. Three replicates per compound per concentration), 5% CO ₂ , 37 ° C incubation for 72 hours. After 72 hours, add 50 μl Cell Titer-Glo to each well, mix on a shaker for 2 min, and incubate for 10 min, then read with EnVision.

data analysis:

The cell activity was plotted with GraphPad Prism version 5, and the curve was fitted with a nonlinear regression model with a sigmoidal dose response, thereby calculating IC ₅₀ data.

Most of the compounds of the present invention have good performance in the cell proliferation inhibition IC _{50 of} BaF3 (EGFR-del19 / T790M / C797S) and the cell proliferation inhibition IC _{50 of} BaF ₃ (EGFR-L858R / T790M / C797S), which is significantly better than The standard compound Brigatinib. The test results are shown in the following table:

In the above, the embodiment of the present invention has been described. However, the present invention is not limited to the above-mentioned embodiment. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (11)

1. The compound represented by formula (I), its stereoisomers, racemates, tautomers, isotope labels, nitrogen oxides or pharmaceutically acceptable salts thereof,

   

   Where E is selected from

   

   Y is selected from O or S; R ₁ , R ₂ , R ₃ , R _{4 are the} same or different, and are independently selected from C _1-12 alkyl;

   A is selected from N or C-Q;

   B is selected from N or C-D;

   Q and D are the same or different and are independently selected from H, halogen, cyano, hydroxy, amino, unsubstituted or optionally substituted with one, two or more R _a of the following groups: C _1-12 alkane Group, C _1-12 heteroalkyl, C _2-12 alkenyl, C _2-12 heteroalkenyl, C _2-12 alkynyl, C _2-12 heteroalkynyl, C _1-12 alkoxy, C _{3 -20} cycloalkyl, 3-20 membered heterocyclic group, C _6-20 aryl group, 5-20 membered heteroaryl group, -COOC _1-12 alkyl group, -COC _1-12 alkyl group;

   Each R _{a is the} same or different and is independently selected from halogen, cyano, amino, hydroxy, unsubstituted or optionally substituted with one, two or more Rs of the following groups: C _1-12 alkyl, C _1-12 heteroalkyl, C _1-12 alkoxy, C _3-20 cycloalkyl, 3-20 membered heterocyclyl, -COOC _1-12 alkyl, -COC _1-12 alkyl;

   V and Z are the same or different and are independently selected from H, halogen, cyano, hydroxy, amino, unsubstituted or optionally substituted with one, two or more R _b of the following groups: C _1-12 alkane Group, C _1-12 heteroalkyl, C _2-12 alkenyl, C _2-12 heteroalkenyl, C _2-12 alkynyl, C _2-12 heteroalkynyl, C _1-12 alkoxy, C _{3 -20} cycloalkyl, 3-20 membered heterocyclic group, C _6-20 aryl group, 5-20 membered heteroaryl group, -COOC _1-12 alkyl group, -COC _1-12 alkyl group, -N (C _{1 -12} alkyl) ₂ , -NHC _1-12 alkyl;

   Each R _{b is the} same or different and is independently selected from halogen, cyano, hydroxy, amino, unsubstituted or optionally substituted with one, two or more Rs of the following groups: C _1-12 alkyl, C _1-12 alkoxy, C _1-12 heteroalkyl, C _3-20 cycloalkyl, 3-20 membered heterocyclyl;

   Each Rs is the same or different and is independently selected from halogen, CN, OH, C _1-12 alkyl;

   W is selected from the group consisting of hydrogen, unsubstituted or optionally substituted with one, two or more R _c : C _1-12 alkyl, C _1-12 heteroalkyl, C _1-12 alkanoyl, C _1-12 heteroalkanoyl, 3-20 membered heterocyclic acyl, C _3-20 cycloalkyl, 3-20 membered heterocyclyl, C _6-18 aryl, 5-20 membered heteroaryl, C _{3-20 Spirocyclic} group formed by cycloalkyl and C _3-20 cycloalkyl, spirocyclic group formed by C _3-20 cycloalkyl and 3-20 membered heterocyclic group, 3-20 membered heterocyclic group and 3-20 membered Spirocyclic group formed by heterocyclic group, C3-20 cycloalkyl and C3-20 cycloalkyl cyclic group, C3-20 cycloalkyl and 3-20 membered heterocyclic group Cyclocyclic group, 3-20 member A heterocyclic group and a 3-20 membered heterocyclic group, a C _5-14 bridged cyclic group, a 5-14 membered heterocyclic ring group;

   Each R _{c is the} same or different, and is independently selected from the group consisting of = O, halogen, cyano, hydroxy, amino, carboxy, unsubstituted or optionally substituted with one, two or more _Rd : C _1-12 alkyl, C _1-12 alkoxy, C _1-12 heteroalkyl, -S (O) ₂ C _1-12 alkyl, -C (O) ₂ C _1-12 alkyl, -COC _1-12 alkyl, C _3-20 cycloalkyl, 3-20 membered heterocyclyl;

   Each _{Rd is the} same or different, and is independently selected from halogen, = O, halogen, cyano, hydroxyl, -S (O) ₂ C _1-12 alkyl, -COOC _1-12 alkyl, -COC _{1- 12} alkyl, C _1-12 alkyl; when W is selected from 3-20 membered heterocyclic group, C _3-20 cycloalkyl and 3-20 membered heterocyclic group formed spiro ring group, 3-20 membered heterocyclic ring When the group is a spiro ring group formed from a 3-20 membered heterocyclic group and a 5-14 membered heterobridged ring group, the substituted R _c may be connected by a hetero atom or a carbon atom.
2. The compound represented by formula (I) according to claim 1, its stereoisomer, racemate, tautomer, isotope label, nitrogen oxide, or a pharmaceutically acceptable salt thereof, characterized in that ,

   Each R _a , R _b , R _{c is the} same or different, and is independently selected from -F, -Cl, -Br, -I, -OH, -CN, -NH ₂ , -CH ₃ , CH ₃ CH _2- , -CF ₃ , -CHF ₂ , -CH ₂ F, NH ₂ CH _2- , -NH (CH ₃ ) ₂ , -OCH ₃ , CH ₃ CH ₂ O-, CH ₃ OCH _2- , MeSO ₂ -,- CH ₂ CH ₂ F, -CH ₂ CHF ₂ , -CH ₂ C (CH ₃ ) ₂ OH, -CH ₂ CH ₂ SO ₂ Me, -CH ₂ OH, -CH ₂ CH ₂ OH, -CH ₂ CH ₂ CN , -CH ₂ CH ₂ OCH ₃ , -COOCH (CF ₃ ) ₂ , -COCH ₂ OH, -CH ₂ COOH, -CH ₂ CONH ₂ , 2- (1-morpholinyl) ethyl, 1-methylpiper Pyridin-4-acyl, 3-pyridyl, 2- (pyrrolidin-1-yl) ethyl, 3-oxetanyl, tetrahydropyrrolyl, morpholinyl, 4-methyl-1-piperazine Group, 1-methyl-4-piperidinyl;

   Each Rs is the same or different and is independently selected from halogen, CN, OH, C _1-6 alkyl;

   E is selected from

   

   Y is selected from O or S; R ₁ , R ₂ , R ₃ , and R _{4 are the} same or different, and are independently selected from C _1-6 alkyl; A is selected from N or CQ; B is selected from N or CD; A and At least one of B is N, and A and B are not N at the same time;

   Q and D are the same or different and are independently selected from H, halogen, cyano, C _1-6 alkyl, halogen substituted C _1-6 alkyl, C _1-6 heteroalkyl, C _1-6 alkoxy Group, C _3-12 cycloalkyl, 3-12 membered heterocyclic group, -COOC _1-6 alkyl, -COC _1-6 alkyl;

   V and Z are the same or different, and are independently selected from H, halogen, cyano, C _1-6 alkyl, C _1-6 alkoxy, C _3-6 cycloalkyl;

   W is selected from the following groups which are unsubstituted or optionally substituted with one, two or more R _c : C _1-6 alkyl, C _1-6 heteroalkyl, C _3-12 cycloalkyl, 3- 12-membered heterocyclic group, C _6-12 aryl group, 5-12 membered heteroaryl group, C _3-6 cycloalkyl and C _3-6 cycloalkyl formed spiro ring group, C _3-12 cycloalkyl and The spirocyclic group formed by the 3-12 membered heterocyclic group, the spirocyclic group formed by the 3-12 membered heterocyclic group and the 3-12 membered heterocyclic group, the combination of the C _3-6 cycloalkyl group and the C3-6 heterocyclic group Cyclic group, C3-6 heterocyclic group and C3-6 heterocyclic group fused ring group, C _5-14 bridge ring group, 5-14 member heterobridge ring group.
3. The compound represented by formula (I) according to claim 1, its stereoisomer, racemate, tautomer, isotope label, nitrogen oxide, or a pharmaceutically acceptable salt thereof, characterized in that , E is selected from

   

   Y is selected from O or S;

   At least one of A and B is N, and A and B are not N at the same time;

   Q and D are the same or different, and are independently selected from H, F, Cl, Br, trifluoromethyl, cyano, isopropyl, methoxy, cyclopropyl, -COOC ₂ H ₅ ;

   V and Z are the same or different, and are independently selected from H, F, Cl, Br, methyl, methoxy, cyclopropyl;

   W is selected from: tert-butyl,

   

   
4. The compound represented by formula (I) according to claim 1, a stereoisomer, a racemate, a tautomer, an isotopic label, a nitrogen oxide, or a pharmaceutically acceptable salt thereof, characterized in that The compound of formula (I) is selected from the following compounds or pharmaceutically acceptable salts thereof:

   

   

   

   

   

   

   

   

   
5. The compound represented by formula (I) according to claim 1, its stereoisomer, racemate, tautomer, isotope label, nitrogen oxide, or a pharmaceutically acceptable salt thereof, characterized in that The salt is hydrochloride, trifluoroacetate or formate.
6. A pharmaceutical composition, characterized in that it contains a therapeutically effective amount of the compound represented by any one of claims 1-5, its stereoisomer, racemate, tautomer, At least one of an isotope label, nitrogen oxide, or a pharmaceutically acceptable salt thereof.
7. The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.
8. The compound represented by formula (I) according to any one of claims 1 to 5, its stereoisomers, racemates, tautomers, isotopic labels, nitrogen oxides or pharmaceutically acceptable salts thereof Use of at least one of them in the preparation of a medicament for treating cancer.
9. The use according to claim 8, characterized in that the cancer is non-small cell lung cancer, glioblastoma, pancreatic cancer, head and neck cancer, breast cancer, colorectal cancer, epithelial cancer, ovarian cancer, prostate cancer, Adenocarcinoma or squamous cell carcinoma.
10. The compound represented by formula (I) according to any one of claims 1 to 5, its stereoisomer, racemate, tautomer, isotopic label, nitrogen oxide, or a pharmaceutically acceptable Use of at least one of the salts in the preparation of a medicament for treating diseases caused by EGFR mutation.
11. The use according to claim 10, characterized in that the EGFR mutation is one, two or more mutations selected from: (1) Del19; (2) T790M; (3) C797S; (4) L858R ; (5) T790M; (6) C797S.