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CN112771032B - Pyrimidine pyrazole compounds as fourth generation EGFR inhibitors - Google Patents

Pyrimidine pyrazole compounds as fourth generation EGFR inhibitors Download PDF

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CN112771032B
CN112771032B CN201980061241.8A CN201980061241A CN112771032B CN 112771032 B CN112771032 B CN 112771032B CN 201980061241 A CN201980061241 A CN 201980061241A CN 112771032 B CN112771032 B CN 112771032B
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CN112771032A (en
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桑迎霞
谷晓辉
诸舜伟
薛黎婷
于晓虹
任晋生
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    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
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Abstract

Novel pyrimidine pyrazole compounds of fourth generation (T790M/C797S mutation) EGFR kinase inhibitors and medical application thereof are disclosed, and particularly, compounds shown as a formula (I), stereoisomers, racemates, tautomers, isotopic labels, nitric oxides or pharmaceutically acceptable salts thereof are disclosed. The compound has better curative effect on diseases caused by EGFR Del19/T790M/C797S and L858R/T790M/C797S abnormal mutation.

Description

Pyrimidine pyrazole compounds as fourth generation EGFR inhibitors
The application claims the priority of two prior applications, namely a patent application number of 201811266972.X, the invention name of pyrimidine pyrazole compound as a fourth generation EGFR inhibitor, which is submitted to the Chinese intellectual property office in 2018, 10, 29 and 2019, 6, 4, the patent application number of 201910479288.8, the invention name of pyrimidine pyrazole compound as a fourth generation EGFR inhibitor. The entire disclosures of both applications are incorporated by reference into this application.
Technical Field
The invention relates to a novel pyrimidine pyrazole compound serving as a fourth-generation (T790M/C797S mutation) EGFR kinase inhibitor and medical application thereof, and particularly discloses a compound shown as a formula (I), a stereoisomer, a racemate, a tautomer, an isotope label, a nitric oxide or a pharmaceutically acceptable salt thereof.
Background
EGFR (Epidermal Growth Factor Receptor) is a Receptor for cell proliferation and signaling of the Epidermal Growth Factor (EGF). EGFR belongs to the ErbB receptor family, which includes EGFR (ErbB-1), HER2/c-neu (ErbB-2), HER 3 (ErbB-3), and HER4 (ErbB-4). EGFR is also known as HER1, erbB1. The EGFR is widely distributed on the cell surfaces of epithelial cells, fibroblasts, glial cells, keratinocytes and the like of mammals, and an EGFR signal channel plays an important role in the physiological processes of growth, proliferation, differentiation and the like of cells. EGFR is divided into three regions: an extracellular ligand binding domain, a transmembrane domain, and an intracellular kinase domain. After being bound by corresponding ligands, EGFR can be induced to form homo-or heterodimers, so that an intracellular tyrosine kinase pathway is activated, and the EGFR can be self-phosphorylated, thereby guiding downstream phosphorylation including MAPK, akt and JNK pathways and inducing cell proliferation.
An EGFR Tyrosine Kinase Inhibitor (TKI) inhibits receptor phosphorylation and activation of downstream signal transduction molecules by blocking binding of endogenous ATP to an intracellular Kinase domain, thereby blocking proliferation of tumor cells. Although EGFR targeted therapies have been successfully advanced into the clinical stage and drugs are already on the market, genetic mutations in EGFR have led to drug resistance. The mutation mainly occurs in exons 18-21, wherein deletion of exon 19 and point mutation of L858R in exon 21 are the most common mutant subtypes, accounting for 90% of all mutant types. With the development and use of drugs, most of the resistance occurs in the secondary mutation of the gatekeeper region T790M of the kinase. The third-generation irreversible inhibitor developed in recent years has good inhibitory activity on T790M mutation, but C797S mutation is also inevitable and is the main drug resistance mechanism (about 40%) of the star drug AZD 9291. AZD9291 has already entered first-line treatment in 2018, and there is an urgent need to develop novel, safer and more effective EGFR TKIs against C797S mutation.
Disclosure of Invention
In order to specifically solve the problem of drug resistance of the third-generation inhibitor, namely the problem of third mutation C797S of EGFR, the invention provides a compound shown as a formula (I), a stereoisomer, a racemate, a tautomer, an isotopic marker, a nitrogen oxide or a pharmaceutically acceptable salt thereof,
Figure GDA0003667177710000011
wherein E is selected from
Figure GDA0003667177710000021
Y is selected from O or S; r 1 、R 2 、R 3 、R 4 Identical or different, independently of one another, from C 1-12 An alkyl group;
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, unsubstitutedSubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-12 Alkyl radical, C 1-12 Heteroalkyl group, C 2-12 Alkenyl radical, C 2-12 Heteroalkenyl, C 2-12 Alkynyl, C 2-12 Heteroalkynyl, C 1-12 Alkoxy radical, C 3-20 Cycloalkyl, 3-20 membered heterocyclyl, C 6-20 Aryl, 5-20 membered heteroaryl, -COOC 1-12 Alkyl, -COC 1-12 An alkyl group;
each R a Identical or different, independently of one another, from the group consisting of halogen, cyano, amino, hydroxyl, the following groups which are unsubstituted or optionally substituted by one, two or more Rs: c 1-12 Alkyl radical, C 1-12 Heteroalkyl group, C 1-12 Alkoxy radical, C 3-20 Cycloalkyl, 3-20 membered heterocyclyl, -COOC 1-12 Alkyl, -COC 1-12 An alkyl group;
v and Z are identical or different and are independently selected from H, halogen, cyano, hydroxy, amino, unsubstituted or optionally substituted by one, two or more R b Substituted of the following groups: c 1-12 Alkyl radical, C 1-12 Heteroalkyl group, C 2-12 Alkenyl radical, C 2-12 Heteroalkenyl, C 2-12 Alkynyl, C 2-12 Heteroalkynyl, C 1-12 Alkoxy radical, 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 radical) 2 、-NHC 1-12 An alkyl group;
each R b Identical or different, independently of one another, from the group consisting of halogen, cyano, hydroxy, amino, the following groups which are unsubstituted or optionally substituted by one, two or more Rs: c 1-12 Alkyl radical, C 1-12 Alkoxy radical, C 1-12 Heteroalkyl group, C 3-20 Cycloalkyl, 3-20 membered heterocyclyl;
each Rs, which are identical or different, is independently selected from halogen, CN, OH, C 1-12 An alkyl group;
w is selected from hydrogen, unsubstituted or optionally substituted by one, two or more R c Substituted of the following groups: c 1-12 Alkyl radical, C 1-12 Heteroalkyl group, C 1-12 Alkanoyl radical, C 1-12 Heteroalkanoyl, 3-20 membered heterocycloyl, C 3-20 Cycloalkyl, 3-20 membered heterocyclyl, C 6-18 Aryl, 5-20 membered heteroaryl, C 3-20 Cycloalkyl radicals and C 3-20 Spiro ring radical formed by cycloalkyl, C 3-20 A spiro ring group formed by the cycloalkyl and the 3-20 membered heterocyclic group, a spiro ring group formed by the 3-20 membered heterocyclic group and the 3-20 membered heterocyclic group, a combined ring group of the C3-20 cycloalkyl and the C3-20 cycloalkyl, a combined ring group of the C3-20 cycloalkyl and the 3-20 membered heterocyclic group, a combined ring group of the 3-20 membered heterocyclic group and the 3-20 membered heterocyclic group, a C 5-14 Bridged ring group, 5-14 membered heterobridged ring group;
each R c Identical or different, independently from each other, selected from = O, halogen, cyano, hydroxy, amino, carboxy, unsubstituted or optionally substituted by one, two or more R d Substituted of the following groups: c 1-12 Alkyl radical, C 1-12 Alkoxy radical, C 1-12 Heteroalkyl, -S (O) 2 C 1-12 Alkyl, -C (O) 2 C 1-12 Alkyl, -COC 1-12 Alkyl radical, C 3-20 Cycloalkyl, 3-20 membered heterocyclyl;
each R d Identical or different, independently of one another, from the group consisting of halogen, = O, halogen, cyano, hydroxy, -S (O) 2 C 1-12 Alkyl, -COOC 1-12 Alkyl, -COC 1-12 Alkyl radical, C 1-12 An alkyl group; when W is selected from 3-20 membered heterocyclic group, C 3-20 When the substituted R is a spiro ring group formed by a cycloalkyl group and a 3-20 membered heterocyclic group, a spiro ring group formed by a 3-20 membered heterocyclic group and a 3-20 membered heterocyclic group, or a 5-14 membered hetero bridge ring group c May be attached by heteroatoms or carbon atoms.
According to an embodiment of the invention, Q and D are identical or different and are independently from each other selected from H, halogen, cyano, hydroxy, amino, unsubstituted or optionally substituted by one, two or more R a Substituted of the following groups: c 1-6 Alkyl radical, C 1-6 Heteroalkyl group, C 1-6 Alkenyl radical, C 1-6 Heteroalkenyl, C 1-6 Alkynyl, C 1-6 Heteroalkynyl, C 3-6 Cycloalkyl radical, C 3-6 Heterocycloalkyl, phenyl and 5-6 membered heteroaryl;
v and Z are identical or different and are independently selected from H, halogen, cyano, hydroxy, amino, unsubstituted or optionally substituted by one, two or more R b Substituted of the following groups: c 1-6 Alkyl radical, C 1-6 Heteroalkyl group, C 3-6 Cycloalkyl radical, C 3-6 Heterocycloalkyl, phenyl and 5-to 6-membered heteroaryl, said C 1-6 Alkyl radical, C 1-6 Heteroalkyl group, C 3-6 Cycloalkyl radical, C 3-6 Heterocycloalkyl, phenyl and 5-6 membered heteroaryl;
w is selected from hydrogen, unsubstituted or optionally substituted by one, two or more R c Substituted of the following groups: c 1-6 Alkyl radical, C 1-6 Heteroalkyl group, C 1-3 Alkanoyl radical, C 1-3 Heteroalkanoyl, C 3-8 Cycloalkyl radical, C 3-8 Heterocyclic group, C 3-6 Cycloalkyl radicals and C 3-6 Spiro ring radical of cycloalkyl, C 3-6 Cycloalkyl radicals and C 3-6 Spiro ring group of heterocyclic group of (1), C 3-6 Cycloalkyl radicals and C 3-6 Cycloalkyl fused ring radical, C 3-6 Cycloalkyl radicals and C 3-6 Heterocyclic radical, C 3-6 Heterocyclyl and C 3-6 Heterocyclic radical, C 5-14 Bridge ring radical, C 5-14 A heterobridged ring group;
according to an embodiment of the invention, each R is a 、R b 、R c The same or different, or a combination thereof, independently of one another, from the group consisting of-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-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 invention, each Rs, equal or different, is independently selected from halogen, CN, OH, C 1-6 An alkyl group;
preferably, E is selected from
Figure GDA0003667177710000031
Y is selected from O or S; r 1 、R 2 、R 3 、R 4 Identical or different, independently of one another, from C 1-6 An alkyl group; a is selected from N or C-Q; b is selected from N or C-D; at least one of A and B is N, and A and B are not N at the same time;
q and D are identical or different and are independently selected from H, halogen, cyano, C 1-6 Alkyl, halogen substituted C 1-6 Alkyl radical, C 1-6 Heteroalkyl group, C 1-6 Alkoxy radical, C 3-12 Cycloalkyl, 3-12 membered heterocyclyl, -COOC 1-6 Alkyl, -COC 1-6 An alkyl group;
v and Z are identical or different and are independently selected from H, halogen, cyano, C 1-6 Alkyl radical, C 1-6 Alkoxy radical, C 3-6 A cycloalkyl group;
w is selected from unsubstituted or optionally substituted by one, two or more R c Substituted of the following groups: c 1-6 Alkyl radical, C 1-6 Heteroalkyl group, C 3-12 Cycloalkyl, 3-12 membered heterocyclyl, C 6-12 Aryl, 5-12 membered heteroaryl, C 3-6 Cycloalkyl radicals and C 3-6 Spiro ring radical formed by cycloalkyl, C 3-12 Spiro ring group formed by cycloalkyl and 3-12 membered heterocyclic group, spiro ring group formed by 3-12 membered heterocyclic group and 3-12 membered heterocyclic group, C 3-6 Cyclic group of cycloalkyl and C3-6 heterocyclic group, cyclic group of C3-6 heterocyclic group and C3-6 heterocyclic group, C 5-14 Bridged ring group, 5-14 membered heterobridged ring group;
according to an embodiment of the invention, each R is a 、R b Identical or different, independently of one another, from the group consisting of Rr、-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, 1-tetrahydropyrrolyl, 2- (1-morpholinyl) ethyl, 1-methylpiperidin-4-acyl, 3-pyridyl, 2- (pyrrolidin-1-yl) ethyl;
more preferably, E is selected from
Figure GDA0003667177710000032
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 identical or different and are each independently selected from H, F, cl, br, trifluoromethyl, cyano, isopropyl, methoxy, cyclopropyl, -COOC 2 H 5 ；
V and Z are identical or different and are independently selected from H, F, cl, br, methyl, methoxy, cyclopropyl;
w is selected from: a tert-butyl group,
Figure GDA0003667177710000033
Figure GDA0003667177710000041
In some preferred embodiments, when W is
Figure GDA0003667177710000042
Z is methyl, V is H, B is N, E is
Figure GDA0003667177710000043
When Y is O, A is C-D, D is selected from: halogen, C1-6 alkoxy, C3-6 cycloalkyl; when W is
Figure GDA0003667177710000051
Z is methoxy, V is H, B is N, E is
Figure GDA0003667177710000052
When Y is O, A is C-D, D is selected from: c1-6 alkyl, halogen substituted C1-6 alkyl, C1-6 alkoxy, C3-6 cycloalkyl, C1-6 heteroalkyl; when W is
Figure GDA0003667177710000053
Z is methyl, V is H, A is N, E is
Figure GDA0003667177710000054
When Y is O, B is C-Q, 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:
Figure GDA0003667177710000055
Figure GDA0003667177710000061
Figure GDA0003667177710000071
Figure GDA0003667177710000081
Figure GDA0003667177710000091
Figure GDA0003667177710000101
Figure GDA0003667177710000111
Figure GDA0003667177710000121
Figure GDA0003667177710000131
wherein the salt is hydrochloride, trifluoroacetate or formate.
The invention also provides a pharmaceutical composition, which comprises at least one of a therapeutically effective amount of a compound shown in the formula (I), a stereoisomer, a racemate, a tautomer, an isotopic marker, a nitrogen oxide or a pharmaceutically acceptable salt thereof.
Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.
The invention also provides application of at least one of the compound shown in the formula (I), a stereoisomer, a racemate, a tautomer, an isotopic marker, a nitrogen oxide or a pharmaceutically acceptable salt thereof in preparing a medicament for treating 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 invention also provides application of at least one of the compounds shown in the formula (I), stereoisomers, racemates, tautomers, isotopic markers, nitric oxides or pharmaceutically acceptable salts thereof in preparing a medicament 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; and (6) C797S.
The present invention also provides a method of treating cancer comprising: administering at least one of a compound of formula (I), a stereoisomer, racemate, tautomer, isotopic label, nitroxide, or pharmaceutically acceptable salt thereof, to a subject in need thereof.
Has the advantages that: in vitro activity experiment results prove that the compound has better potential curative effect on diseases caused by EGFR Del19/T790M/C797S and L858R/T790M/C797S abnormal mutation.
Definition and description of terms
In the invention
Figure GDA0003667177710000132
And (b) represents a connection site.
As used herein, the following terms and phrases are intended to have the following meanings, unless otherwise indicated. A particular term or phrase, unless specifically defined, should not be considered as indefinite or unclear, but rather construed according to ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commodity or its active ingredient.
Unless otherwise indicated, the definitions of groups and terms described in the specification and claims of the present application, including definitions thereof as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. The definitions of the groups and the structures of the compounds in such combinations and after the combination are within the scope of the present specification.
Wherein "more" means three or more.
The term "halogen" refers to F, cl, br and I. In other words, F, cl, br, and I may be described as "halogen" in the present specification.
The term "C 1-12 Alkyl "is understood to preferably mean a straight-chain or branched, saturated, monovalent hydrocarbon radical having from 1 to 12 carbon atoms, preferably C 1-10 An alkyl group. "C 1-10 Alkyl "is understood to preferably mean a straight-chain or branched, saturated monovalent hydrocarbon radical 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-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, or the like or isomers thereof. In particular, the radicals have 1,2, 3,4, 5,6 carbon atoms ("C) 1-6 Alkyl groups) such as methyl, ethyl, propyl, butyl, isopropyl, isobutyl, sec-butyl, tert-butyl, more particularly groups having 1,2 or 3 carbon atoms ("C) 1-3 Alkyl groups) such as methyl, ethyl, n-propyl or isopropyl.
The term "C 2-12 Alkenyl "is understood to preferably mean a straight-chain or branched monovalent hydrocarbon radical comprising one or more double bonds and having from 2 to 12 carbon atoms, preferably" C 2-10 Alkenyl group ", more preferably" C 2-6 Alkenyl ". "C 2-10 Alkenyl "is understood to preferably mean a straight-chain or branched monovalent hydrocarbon radical which comprises one or more double bonds and has 2,3, 4,5, 6, 7, 8, 9 or 10 carbon atoms, in particular 2 or 3 carbon atoms (" C) 2-3 Alkenyl "), it being understood that in the case where the alkenyl group comprises more than one double bond, the double bonds may be separated from each other or conjugated. Alkenyl 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-enylA radical, (Z) -pent-2-enyl, (E) -pent-1-enyl, (Z) -pent-1-enyl, hex-5-enyl, (E) -hex-4-enyl, (Z) -hex-4-enyl, (E) -hex-3-enyl, (Z) -hex-3-enyl, (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-methylbut-2-enyl, (E) -2-methylbut-2-enyl, (Z) -2-methylbut-2-enyl, (E) -1-methyl-2-enyl, (Z) -1-methylbut-2-enyl, (Z) -1-methyl-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-methylbut-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl. As will be understood by those skilled in the art, the term "C 1-n Alkenyl radicals "(e.g." C) 1-12 Alkenyl "or" C 1-6 Alkenyl ") is identical to the above-mentioned" C 2-n Definition of "alkenyl" (e.g. C) 2-12 Alkenyl).
The term "C 2-12 Alkynyl "is understood to mean a straight-chain or branched, monovalent hydrocarbon radical comprising one or more triple bonds and having from 2 to 12 carbon atoms, preferably" C 2 -C 10 Alkynyl group ", more preferably" C 2 -C 6 Alkynyl ". The term "C 2 -C 10 Alkynyl "is understood as preferably meaning a straight-chain or branched, monovalent hydrocarbon radical which contains one or more triple bonds and has 2,3, 4,5, 6, 7, 8, 9 or 10 carbon atoms, in particular 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-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl2-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-2-ynyl, 1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl, 2,2-dimethylbut-3-ynyl, 1,1-dimethylbut-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. As will be understood by those skilled in the art, the term "C 1-n Alkynyl radicals "(e.g." C) 1-12 Alkynyl "or" C 1-6 Alkynyl ") is identical to the above-mentioned" C 2-n Definition of alkynyl (e.g. C) 2-12 Alkynyl).
"hetero" represents a heteroatom or a heteroatom group, each independently selected from-O-, -S-, = O, = S, -O-N =, -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, the number of heteroatoms or groups of heteroatoms is selected from 1,2 or 3, respectively.
As will be understood by those skilled in the art, the foregoing term "C" is intended to mean 1-12 Alkyl group "," C 2-12 Alkenyl group "," C 2-12 Alkynyl "definition of the radicals likewise applies to" C 1-12 Heteroalkyl group "," C 2-12 Heteroalkenyl "," C 2-12 Heteroalkynyl "refers to alkyl, alkenyl, and alkynyl moieties.
The term "C 3-20 Cycloalkyl "is understood to mean a saturated or unsaturated, monovalent, monocyclic or bicyclic hydrocarbon ring having from 3 to 20 carbon atoms, preferably" C 3-10 Cycloalkyl groups ". 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. Said C is 3-10 Cycloalkyl groups may be monocyclic hydrocarbon groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or bicyclic hydrocarbon groups such as decalin rings. Said "C 3-20 Cycloalkyl "may be further optionally substituted by, for example, C 1-8 Cycloalkyl radical, C 3-8 A cycloalkyl group, a,C 3-6 Cycloalkyl, and the like.
The term "3-20 membered heterocyclyl" means a saturated or unsaturated monovalent monocyclic or bicyclic hydrocarbon ring comprising 1-5 heteroatoms independently selected from N, O and S, preferably "3-10 membered heterocyclyl". The term "3-10 membered heterocyclyl" means a saturated monovalent monocyclic or bicyclic hydrocarbon ring comprising 1-5, preferably 1-3 heteroatoms selected from N, O and S. The heterocyclic group may be attached 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: 4-membered rings such as azetidinyl, oxetanyl; 5-membered rings such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or a 7-membered ring such as diazepanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclyl group may be bicyclic, for example but not limited to a 5,5 membered ring, such as a hexahydrocyclopenta [ c ] pyrrol-2 (1H) -yl ring, or a 5,6 membered bicyclic ring, such as a hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl ring. The nitrogen atom containing ring may be partially unsaturated, i.e. it may contain one or more double bonds, such as but not limited to 2,5-dihydro-1H-pyrrolyl, 4H- [1,3,4] thiadiazinyl, 4,5-dihydrooxazolyl, or 4H- [1,4] thiazinyl, or it may be benzo-fused, such as but not limited to dihydroisoquinolinyl. According to the invention, the heterocyclic radical is non-aromatic.
The terms "C3-6 heterocycloalkyl", "C3-6 heterocyclyl" correspond to a heterocyclyl group having 3-6C atoms and are also included within the above-mentioned "3-20 membered heterocyclyl".
The term "C 6-20 Aryl "is to be understood as preferably meaning a mono-, bi-or tricyclic hydrocarbon ring of monovalent or partially aromatic character having from 6 to 20 carbon atoms, preferably" C 6-14 Aryl ". The term "C 6-14 Aryl "is to be understood as preferably meaning a mono-, bi-or tricyclic hydrocarbon ring having a monovalent or partially aromatic character with 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms (" C 6-14 Aryl group "), in particular a ring having 6 carbon atoms (" C 6 Aryl "), such as phenyl; or biphenyl, or is a ring having 9 carbon atoms ("C 9 Aryl), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C 10 Aryl radicals), such as tetralinyl, dihydronaphthyl or naphthyl, or rings having 13 carbon atoms ("C 13 Aryl radicals), such as the fluorenyl radical, or a ring having 14 carbon atoms ("C) 14 Aryl), such as anthracenyl.
The term "5-20 membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: having 5 to 20 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, e.g., "5-14 membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: which have 5,6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and which contain 1 to 5, preferably 1 to 3, heteroatoms each independently selected from N, O and S and, in addition, can be benzo-fused in each case. In particular, heteroaryl is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, thia-4H-pyrazolyl and the like and their benzo derivatives, such as benzofuryl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl, isoquinolyl, and the like; or azocinyl, indolizinyl, purinyl and the like and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and the like.
Unless otherwise specified, heterocyclyl, heteroaryl or heteroarylene include all possible isomeric forms thereof, e.g., positional isomers thereof. Thus, for some illustrative, non-limiting examples, pyridyl or pyridinylene includes pyridin-2-yl, pyridinylene-2-yl, pyridin-3-yl, pyridinylene-3-yl, pyridin-4-yl, and pyridinylene-4-yl; thienyl or thienylene includes thien-2-yl, thien-3-yl and thien-3-yl.
The term "C 5-14 Bridged ring groups "refer to 5-to 14-membered all-carbon polycyclic groups in which any two rings share two carbon atoms not directly attached, and these may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. Comprises 7 to 10 yuan. For example the following groups:
Figure GDA0003667177710000151
they can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl groups depending on the number of constituent rings.
The term "C 5-14 Heterobridged cyclic "refers to 5-to 14-membered polycyclic groups in which any two rings share two atoms not directly connected, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system, wherein one or more ring atoms are selected from nitrogen, oxygen, or heteroatoms of S (O) n (where n is an integer from 0 to 2), and the remaining ring atoms are carbon. Preferably 7 to 10 membered. For example:
Figure GDA0003667177710000161
they may be classified into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups according to the number of constituent rings, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic.
"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.
The term "substituted" means that any one or more hydrogen atoms on the specified atom is replaced with a substituent, and may include variations of deuterium and hydrogen, as long as the valence of the specified atom is normal and the substituted compound is stable. When the substituent is keto (i.e = O), it means that two hydrogen atoms are substituted. The keto substitution does not occur on the aromatic group. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the kind of substituent and the tree may be arbitrary on the basis of chemical feasibility.
When any variable (e.g., R) occurs more than one time in the composition or structure of a compound, its definition in each case is independent.
The term "pharmaceutically acceptable" as used herein is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
"pharmaceutically acceptable salts" refers to those salts that retain the biological effectiveness and properties of the parent compound. Such salts include: (1) Salts with acids are obtained by reaction of the free base of the parent compound with inorganic acids including hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid, perchloric acid and the like, or with organic acids including acetic acid, trifluoroacetic acid, propionic acid, acrylic acid, caproic acid, cyclopentanepropionic 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, dodecylsulfuric acid, gluconic acid, glutamic acid, aspartic acid, stearic acid, mandelic acid, succinic acid or malonic acid and the like. (2) The acidic proton present in the parent compound is replaced by a metal ion such as an alkali metal ion, an alkaline earth metal ion or an aluminum ion, or is complexed with an organic base such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, etc.
In addition to salt forms, the compounds provided herein also exist in prodrug forms. Prodrugs of the compounds described herein readily undergo chemical changes under favorable conditions to convert to the compounds of the invention. In addition, prodrugs can be converted to the compounds of the present invention in vivo by chemical or biochemical means.
Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention.
Using wedge and virtual keys, unless otherwise indicated
Figure GDA0003667177710000162
Showing the absolute configuration of a stereocentre, by
Figure GDA0003667177710000163
Indicating the relative configuration of a stereocenter. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, they include the E, Z geometric isomers unless otherwise specified. Likewise, all tautomeric forms are included within the scope of the invention.
The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-) -and (+) -enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, as well as racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Substituents such as alkyl groups may have additional asymmetric carbon atoms. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.
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 one enantiomer of a compound of the invention is desired, it may be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, where the molecule contains a basic group (e.g., amino) or an acidic functional group (e.g., carboxylic acid), diastereomeric salts are formed with an appropriate optically active acid or base, followed by resolution by conventional methods known in the art, and recovery of the pure enantiomers. Furthermore, separation of enantiomers and diastereomers is typically accomplished by using chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamate formation from amines).
The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, compounds may be labeled with radioactive isotopes, such as tritium, iodine-125, C-14. All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.
"pharmaceutical composition" refers to the combination of one or more of the compounds of the present invention, or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof, with other chemical ingredients, such as a pharmaceutically acceptable carrier, excipient, or diluent. The purpose of the pharmaceutical composition is to facilitate the administration process to the animal.
"pharmaceutically acceptable carrier" refers to an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to an 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 (e.g., lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic or methacrylic polymers, gelatin, water, polyethylene glycol, propylene glycol, ethylene glycol, castor oil or hydrogenated or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil, and the like.
Excipients refer to inert substances added to a pharmaceutical composition to further facilitate administration of the compound. Examples of excipients include, without limitation, calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.
The aforementioned pharmaceutical compositions may contain, in addition to pharmaceutically acceptable carriers, adjuvants commonly used in pharmacology, such as: antibacterial agents, antifungal agents, antimicrobial agents, shelf-stable agents, hueing agents, solubilizing agents, thickening agents, surfactants, complexing agents, proteins, amino acids, fats, sugars, vitamins, minerals, trace elements, sweeteners, pigments, flavors or combinations thereof, and the like.
The compounds of the present invention may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, their combinations with other chemical synthetic methods to form embodiments and their equivalents well known to those skilled in the art, with preferred embodiments including, but not limited to, the examples of the present invention.
The solvent used in the present invention can be commercially available. The invention employs the following abbreviations:
Figure GDA0003667177710000171
Figure GDA0003667177710000181
the compounds were named manually or by ChemDraw software and the commercially available compounds were given the supplier catalog name.
LCMS gradient general formula: axBy or CxDy (x + y = 100), A, C: water (acidic: containing 0.1% formic acid, basic: containing 0.1% ammonium bicarbonate), B, D: acetonitrile (acidic: containing 0.1% formic acid, basic: containing 0.1% ammonium bicarbonate), gradient conditions: and acetonitrile y-95%. The total elution time is 2-10 minutes.
HPLC gradient formula: axBy or CxDy (x + y = 100), A, C: water (acidic: containing 0.1% trifluoroacetic acid, basic: containing 0.1% ammonium bicarbonate), B, D: acetonitrile (acidic: containing 0.1% trifluoroacetic acid, basic: containing 0.1% ammonium bicarbonate), gradient conditions: and acetonitrile y-95%. The total elution time is typically 16 minutes.
Detailed Description
The invention is described in detail below by way of examples, but is not meant to be limited to any of the disadvantages of the present invention. Having described the invention in detail and having disclosed specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.
Intermediate preparation example 1: preparation of intermediate D1
Figure GDA0003667177710000182
The compound, tert-butyl 4-hydroxypiperidine-1-carboxylate (5.0g, 24.8mmol) and triethylamine (7.54g, 74.5mmol) were dissolved in dichloromethane (50 mL) at room temperature, and methanesulfonyl chloride (3.13g, 27.3mmol) was slowly added dropwise to the reaction solution. After the addition was complete, the reaction mixture was stirred at room temperature for 8 hours. TLC (PE: etOAc =1, rf = 0.6) showed disappearance of starting material and formation of product. The reaction solution was diluted with water (40 mL), and the mixture was extracted with ethyl acetate (350 mL. Times.2). Anhydrous Na for organic phase synthesis 2 SO 4 The filtrate was concentrated under reduced pressure to give the objective crude product D1 (6.37 g, yield: 91.7%, white solid).
1 H NMR(400MHz,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
Figure GDA0003667177710000183
In a 250mL single-neck flask were added compound D1 (3.00g, 10.7 mmol), compound 4-nitropyrazole (1.28g, 11.3 mmol), and K in that order 2 CO 3 (2.97g, 21.5 mmol) and DMF (40 mL). After the reaction system is stirred uniformly, the temperature is raised to 110 ℃ and the reaction is stirred for 1 hour. TLC (PE: etOAc =3, 1, rf = 0.5) showed disappearance of starting material and formation of product. The reaction solution was diluted with water (100 mL), and the mixture was extracted with ethyl acetate (50 mL. Times.2). The organic phases were combined and washed with a saturated sodium chloride solution (100mL X3), the resulting organic solution was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the objective crude product D2 (1.94 g, yield: 61%, white solid).
1 H NMR(400MHz,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
Figure GDA0003667177710000184
Figure GDA0003667177710000191
After dissolving compound D2 (1.00g, 3.37mmol) in THF (10 mL) in a 100mL three-necked flask, the solution was cooled to-78 deg.C, liHMDS (8.76mL, 8.76mmol) was added dropwise to the reaction solution, and after completion of the addition, the reaction solution was stirred at-78 deg.C for 30min, and then a solution of hexachloroethane (1.20g, 5.06mmol) in THF (10 mL) was slowly added to the reaction solution. After the addition, the mixture was stirred at-78 ℃ for 2.5 hours. TLC (PE: etOAc =3, 1, rf = 0.7) showed disappearance of starting material and formation of product. The reaction was quenched with saturated aqueous ammonium chloride (10 mL), the mixture was extracted with ethyl acetate (50 mL. Times.3), the organic phases were combined and dried over anhydrous sodium sulfate, filtered, 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 give the target product D3 (980 mg, yield: 87.5%, white solid).
1 H NMR(400MHz,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
Figure GDA0003667177710000192
Compound D3 (500mg, 1.69mmol), methyl borate (1.01g, 16.9mmol), K 2 CO 3 (699mg，5.06mmol)，Pd(dppf)Cl 2 (123mg, 0.170mmol) was added to 1,4-Dioxane (5 mL) and H 2 O (1 mL) was added to the mixed solvent, and the mixture was replaced with nitrogen gas 3 times. The reaction was warmed to 85 ℃ and stirred for 10 hours, and compound D4 was detected by LCMS and D3 completely disappeared. The reaction mixture was cooled to room temperature, and added to a mixture of water (10 mL) and ethyl acetate (30 mL), and after separating the two phases, the aqueous phase was extracted with ethyl acetate (30 mL. Times.3). The organic phases were combined and dried and concentrated, and the residue was purified by column chromatography (eluent gradient PE: etOAc =100 from 1 to 5:1) to give the desired product D4 (290 mg, yield: 55.4%, white solid).
1 H NMR(400MHz,CD 3 Cl 3 )δ8.09(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
Figure GDA0003667177710000193
Pd/C (100mg, 10%) was added to a solution of Compound D4 (240mg, 0.770 mmol) in ethanol (5 mL) at room temperature. The reaction mixture was evacuated and filled with hydrogen to displace 3 times, then the system was stirred at 60 ℃ under hydrogen atmosphere for 1 hour and LCMS checked for completion of the reaction. The reaction mixture was filtered to remove Pd/C. The filtrate was concentrated under reduced pressure to give crude objective compound D5 (200 mg, yield: 93%, tan solid).
LCMS:Rt:1.178min；MS m/z(ESI):281.3[M+H]。
Intermediate preparation example 6: preparation of intermediate D6
Figure GDA0003667177710000194
Potassium carbonate (8.8g, 64.4 mmol) was added to a solution of the compound 4-nitro-1-pyrazole (3.6g, 32.2mmol) and the compound methanesulfonic acid (4-hexahydropyranyl) ester (5.8g, 32.2mmol) in DMF (50 mL) at room temperature, and then the reaction mixture was heated to 80 ℃ and stirred for reaction overnight. After completion of the reaction, the reaction mixture was diluted with water (300 mL), the mixture was extracted with ethyl acetate (100mL X3), and the organic phase was washed with a saturated aqueous sodium chloride solution (300mL X3); the organic solution was then dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (elution gradient: PE: etOAc = 5:1) to give the objective product D6 (2.7 g, yield: 43%, yellow solid).
Intermediate preparation example 7: preparation of intermediate D7
Figure GDA0003667177710000201
A solution of compound D6 (0.2g, 1.01mmol) in THF (5 mL) was cooled to-78 ℃ and LiHMDS (2mL, 1M) was slowly added dropwise; after the completion of the dropwise addition, the reaction mixture was stirred for 20 minutes, and hexachloroethane (0.288g, 1.22mmol) was then added to the reaction mixture; the mixture was stirred at-78 ℃ for 1 hour. TLC (PE: etOAc =3, 1, rf = 0.7) showed disappearance of starting material and formation of product. The reaction was quenched with saturated aqueous ammonium chloride (10 mL), the mixture was extracted with ethyl acetate (50 mL. Times.3), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the objective crude product D7 (130 mg, yield: 55%, yellow solid).
1 H NMR(400MHz,DMSO-d6)δ：8.50(s,1H),4.70-4.60(m,1H)3.97(d,J＝8.4Hz,2H),3.53-3.47(m,2H),2.01-1.85(m,4H).
Intermediate preparation example 8: preparation of intermediate D8
Figure GDA0003667177710000202
Compound D7 (300mg, 1.5 mmol) and methylboronic acid (448.9mg, 7.5 mmol) were dissolved in dioxane (10 mL) and water (2 mL) under nitrogen, and Pd (dppf) Cl was added to the system 2 (110mg, 0.15mmol) and potassium phosphate (955mg, 4.5mmol); the reaction system is heated to 90 ℃ after being protected by nitrogen and stirred to react for 16 hours under the condition. After the reaction is finished, filtering the reaction solution, washing a filter cake by using ethyl acetate (20 mL), combining organic phases, drying and filtering, and concentrating the filtrate under reduced pressure; the residue was purified by column chromatography (elution gradient: PE: etOAc =10: 1-5:1) to give the objective product D8 (220 mg, yield: 80.3%, light yellow solid).
1 H NMR(400MHz,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
Figure GDA0003667177710000203
In a 100mL single-necked flask, compound D8 (220mg, 1.04mmol), meOH (50 mL), and Pd/C (22mg, 5%) were added in that order. The reaction system was stirred at room temperature under a hydrogen atmosphere for 16 hours. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd/C, and the filtrate was concentrated under reduced pressure to give the objective crude product D9 (180 mg, yield: 96%, brown solid). The crude product was used directly in the next reaction.
LCMS:Rt:0.365min；MS m/z(ESI):182.4[M+H]。
Intermediate preparation example 10: preparation of intermediate D10
Figure GDA0003667177710000204
The compound (2-aminophenyl) dimethylphosphine oxide (0) was added to a 100mL three-necked flask.5g, 3.27mmol), 2,6-dichloro-5- (trifluoromethyl) pyrimidine (847 mg, 3.92mmol), DIEA (1.26g, 9.81mmol), and DMF (10 mL). The mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction mixture was poured into water (50 mL). The mixture was extracted with EtOAc (30mL 2), the organic phases were combined and washed with saturated aqueous sodium chloride (60mL X2), and the resulting organic solution was washed with anhydrous Na 2 SO 4 Drying, filtering, and concentrating the filtrate under reduced pressure. The residue was purified by column chromatography (eluent gradient: PE: etOAc = 5:1-0:1) to give a mixture of the objective product D10 and 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
Figure GDA0003667177710000211
Compound D6 (200mg, 1.0 mmol) was dissolved in MeOH (20 mL) at room temperature, followed by addition of Pd/C (40 mg); the reaction system was replaced with hydrogen for 3 times and then stirred at room temperature for 2h. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd/C, and the filtrate was concentrated under reduced pressure to give the objective crude product D11 (100 mg, yield: 58.8%, yellow oily liquid). The crude product was used directly in the next reaction.
LCMS:Rt:0.273min；MS m/z(ESI):168.4[M+H]。
Intermediate preparation example 12: preparation of intermediate D12
Figure GDA0003667177710000212
The compound (2-aminophenyl) dimethylphosphine oxide (1.0g, 5.0mmol), 2,4,5-trichloropyrimidine (923mg, 5.0mmol), naHCO was reacted at room temperature 3 (1.59g, 15.0 mmol) was dissolved in EtOH/H 2 In O (45mL, 8Overnight. The reaction half of the starting material was checked by LCMS and the mixture was extracted with ethyl acetate (50 mLX). The organic phases were combined and washed with saturated NaCl solution (100 mL) over anhydrous Na 2 SO 4 Drying, filtering, and concentrating the filtrate. The residue was purified by means of a silica gel column (elution gradient: PE: etOAC =1, 10-1:5) to give the objective intermediate D12 (740 mg, yield: 39.7%, white solid).
LCMS:Rt:1.827min；MS m/z(ESI):316.0[M+H]。
Intermediate preparation example 13: preparation of intermediate D13
Figure GDA0003667177710000213
The compound 4-nitropyrazole (3.39g, 30mmol), 2-bromoethanol (3.75g, 30mmol) and K are added into a 100mL three-necked bottle at room temperature 2 CO 3 (4.97g, 36mmol) and CH 3 CN (50 mL). The mixture was heated to 80 ℃ and reacted 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 give a crude aimed product D13 (4.7 g, yield: 100%, white solid) which was used directly in the next step.
LCMS:Rt:0.775min；MS m/z(ESI):158.1[M+H]。
Intermediate preparation example 14: preparation of intermediate D14
Figure GDA0003667177710000214
D13 (4.7g, 30mmol), TBSCl (5.0g, 33mmol), imidazole (2.5g, 36mmol) and dichloromethane (50 mL) were sequentially added to a 250mL single-neck flask at room temperature. The reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction solution was washed with water (40mL X3), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure the residue was purified by column chromatography (eluent gradient: PE: etOAC =20: 1-10) to obtain the objective product D14 (6.7 g, yield:80%, white solid.
LCMS:Rt:1.399min；MS m/z(ESI):272.2[M+H]。
Intermediate preparation example 15: preparation of intermediate D15
Figure GDA0003667177710000221
LiHMDS (25mL, 25mmol) was slowly added to a solution of compound D14 (2.7g, 10mmol) in THF (30 mL) at-60 ℃. After the addition, the reaction solution was stirred at-60 ℃ for 30 minutes, and then C was added in portions 2 Cl 6 (3.0 g, 12mmol). After the addition was complete, the reaction was stirred at-60 ℃ for 3 hours. After completion of the reaction, the reaction mixture was warmed to 0 ℃ and then quenched with saturated aqueous ammonium chloride (20 mL), the two phases were separated, and the aqueous phase was extracted with ethyl acetate (30mL X3). The organic phases were combined, washed with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient: PE: etOAC =20: 1-10.
LCMS:Rt:1.553min；MS m/z(ESI):306.2[M+H]。
1 H NMR(400MHz,DMSO)δ：8.61(s,1H),4.43(t,J＝4.0Hz,2H),4.05(t,J＝4.0Hz,2H),0.84(s,9H),0.00(s,6H).
Intermediate preparation example 16: preparation of intermediate D16
Figure GDA0003667177710000222
The compounds 4-nitropyrazole (2.26g, 20mmol), 1-bromo-2-methoxyethane (2.76g, 20mmol) and K were added to a 100mL single-neck flask at room temperature 2 CO 3 (6.72g, 40mmol) and CH 3 CN (30 mL). The mixture was heated to 80 ℃ and the reaction stirred for 16 hours. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give crude desired product D16 (3.1 g, yield: 91.1%, colorless liquid) which was used directly in the next step.
LCMS:Rt:0.984min；MS m/z(ESI):172.4[M+H]。
Intermediate preparation example 17: preparation of intermediate D17
Figure GDA0003667177710000223
Compound D16 (342.0 mg, 2mmol), meOH (5 mL), and Pd/C (34.2 mg, 10%) were added sequentially at room temperature in a 100mL single-neck flask. The reaction system was stirred at room temperature under a hydrogen atmosphere for 16 hours. The reaction was completed by LCMS detection, the reaction solution was filtered to remove Pd/C, and the filtrate was concentrated under reduced pressure to give crude desired product D17 (284 mg, yield: 100%, yellow liquid). The crude product was used directly in the next reaction.
LCMS:Rt:0.327min；MS m/z(ESI):142.2[M+H]。
Intermediate preparation example 18: preparation of intermediate D18
Figure GDA0003667177710000231
Compound D14 (271mg, 1.0 mmol), meOH (5 mL), and Pd/C (27.1mg, 10%) were added sequentially at room temperature in a 100mL single-necked flask. The reaction system was stirred at room temperature under a hydrogen atmosphere for 16 hours. The reaction was completed by LCMS detection, the reaction solution was filtered to remove Pd/C, and the filtrate was concentrated under reduced pressure to give crude desired product D18 (241 mg, yield: 100%, black liquid). The crude product was used directly in the next reaction.
LCMS:Rt:0.827min；MS m/z(ESI):242.4[M+H]。
Intermediate preparation example 19: preparation of intermediate D19
Figure GDA0003667177710000232
LiHMDS (15mL, 15mmol) was slowly added to a solution of compound D16 (1.71g, 10mmol) in THF (30 mL) at-60 ℃. After the addition, the reaction solution was stirred at-60 ℃ for 30 minutes, and then C was added in portions 2 Cl 6 (4.68g, 20mmol). After the addition was complete, the reaction was stirred at-60 ℃ for 4 hours. After completion of the reaction, the reaction mixture was warmed to 0 ℃ and then quenched with saturated aqueous ammonium chloride (20 mL), the two phases were separated, and the aqueous phase was extracted with ethyl acetate (30mL X3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient: PE: etOAC =10: 1-5:1) to give the objective product D19 (1.8 g, yield: 87.4%, yellow liquid).
LCMS:Rt:1.068min；MS m/z(ESI):206.0[M+H]。
Intermediate preparation example 20: preparation of intermediate D20
Figure GDA0003667177710000233
Compound D19 (205mg, 1.0 mmol), iron powder (280mg, 5.0 mmol), NH 4 Cl (428mg, 8.0mmol) was added to EtOH/H 2 O (4:1, 20 mL), and then the reaction mixture was heated to 90 ℃ to react for 2 hours. The reaction was completely detected by LCMS, the reaction was cooled to room temperature, then saturated aqueous sodium bicarbonate (50 mL) and ethyl acetate (20 mL) were added to the reaction, the mixture was stirred for 5 minutes and filtered; separating two phases of the filtrate, and extracting the water phase with ethyl acetate (20mL X3); the organic phases were combined, washed with a saturated aqueous solution of sodium chloride (50 mL), then dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the objective intermediate D20 (100 mg, yield: 57.1%, yellow liquid) which was used directly in the next step.
LCMS:Rt:0.51min；MS m/z(ESI):176.2[M+H]。
Intermediate preparation example 21: preparation of intermediate D21
Figure GDA0003667177710000234
Compound D12 (200mg, 0.63mmol), and Compound D5 (177mg, 0.63mmol) andNH 4 cl (101mg, 1.9 mmol) was dissolved in EtOH (5 mL) and the reaction was stirred at 100 ℃ for 3 hours. After completion of the reaction by LCMS detection, the reaction liquid 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, rf = 0.3) to obtain the objective compound D21 (180 mg, yield: 50.8%, white solid).
LCMS:Rt:1.42min；MS m/z(ESI):560.3[M+H]。
Intermediate preparation example 22: preparation of intermediate D22
Figure GDA0003667177710000241
A solution of compound D6 (0.2g, 1.01mmol) in THF (5 mL) was cooled to-78 ℃ and LiHMDS (2mL, 1M) was slowly added dropwise; after the completion of the dropwise addition, the reaction mixture was stirred for 20 minutes, and hexachloroethane (0.288g, 1.22mmol) was then added to the reaction mixture; the mixture was stirred at-78 ℃ for 1 hour. TLC (PE: etOAc =3, 1, rf = 0.7) showed disappearance of starting material and formation of product. The reaction was quenched with saturated aqueous ammonium chloride (10 mL), the mixture was extracted with ethyl acetate (50 mL. Times.3), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the objective crude product D22 (130 mg, yield: 55%, yellow solid).
1 H NMR(400MHz,DMSO-d6)δ：8.50(s,1H),4.70-4.60(m,1H)3.97(d,J＝8.4Hz,2H),3.53-3.47(m,2H),2.01-1.85(m,4H).
Intermediate preparation example 23: preparation of intermediate D23
Figure GDA0003667177710000242
Compound D22 (150mg, 0.64mmol), iron powder (180mg, 3.2mmol) and NH were reacted at room temperature 4 Cl (274mg, 5.12mmol) was added to EtOH/H 2 In O (4:1, 20 mL), the reaction system was heated to 90 ℃ for 2h. LCMS detects reaction completion, the reaction is cooled to room temperature and saturated carbonic acid is addedAqueous sodium hydrogen (50 mL) and ethyl acetate (20 mL); the mixture was stirred for 5 minutes and filtered, after separation of the two phases, the aqueous phase was extracted with ethyl acetate (20mL X3), the organic phases were combined and washed with a saturated aqueous sodium chloride solution (50 mL), and then dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the objective intermediate D23 (100 mg, yield: 76.9%, yellow liquid) which was used directly in the next step.
LCMS:Rt:0.47min；MS m/z(ESI):202.1[M+H]。
Intermediate preparation example 24: preparation of intermediate D24
Figure GDA0003667177710000243
Compound D15 (305mg, 1.0 mmol) and methylboronic acid (360.0 mg,6.0 mmol) were dissolved in dioxane (10 mL) and water (2 mL) under nitrogen, and Pd (dppf) Cl was added to the system 2 (73.3mg, 0.1mmol) and potassium carbonate (276mg, 2.0 mmol). The reaction system is heated to 100 ℃ after being protected by nitrogen and stirred to react for 16 hours under the condition. After the reaction is finished, filtering the reaction solution, washing a filter cake by using ethyl acetate (20 mL), combining organic phases, drying and filtering by using anhydrous sodium sulfate, and concentrating the filtrate under reduced pressure; the residue was purified by column chromatography (elution gradient: PE: etOAc =10, 1-5:1) to give the objective product D24 (160 mg, yield: 56.1%, light yellow liquid).
LCMS:Rt:1.863min；MS m/z(ESI):286.4[M+H]。
Intermediate preparation example 25: preparation of intermediate D25
Figure GDA0003667177710000251
In a 100mL single-necked flask, compound D24 (160mg, 0.56mmol), meOH (5 mL), and Pd/C (1695 mg, 10%) were added in that order. The reaction system was stirred at room temperature under a hydrogen atmosphere for 16 hours. The reaction was completed by LCMS detection, the reaction solution was filtered to remove Pd/C, and the filtrate was concentrated under reduced pressure to give crude desired product D25 (143 mg, yield: 100%, yellow liquid). The crude product was used directly in the next reaction.
LCMS:Rt:0.890min；MS m/z(ESI):256.3[M+H]。
Intermediate preparation 26: preparation of intermediate D26
Figure GDA0003667177710000252
Compound D19 (500mg, 2.5 mmol), methylboronic acid (900mg, 15.0mmol) was dissolved in 1,4-dioxane (36 mL) at room temperature, and then K was added dropwise 2 CO 3 (1.04g, 7.5 mmol) in water (4 mL); the reaction system is replaced by argon for three times, and then Pd (dppf) Cl is added 2 (200mg, 0.25mmol), the system was replaced with argon three times, heated to 80 ℃ and stirred overnight. The reaction was complete by LCMS, the reaction was cooled to room temperature, the mixture was diluted with water (100 mL) and extracted with ethyl acetate (30mL X3), the organic phases were combined and washed with saturated aqueous sodium chloride (100 mL), the solution was dried over anhydrous sodium sulfate filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient: PE: ETOAC = 2:1) to give the target intermediate D26 (170mg, yeild.
LCMS:Rt:1.383min；MS m/z(ESI):186.4[M+H]。
Intermediate preparation example 27: preparation of intermediate D27
Figure GDA0003667177710000253
Compound D26 (170mg, 0.92mmol) was dissolved in MeOH (20 mL) at room temperature, followed by Pd/C (30 mg); the reaction system was replaced with hydrogen three times and stirred at room temperature for 2 hours. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd/C, and the filtrate was concentrated under reduced pressure to give the objective 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
Figure GDA0003667177710000254
Compound E7 (390mg, 0.79mmol) and DIETOAC (308mg, 2.36mmol) were dissolved in DMF (15 mL) at room temperature, methyl bromoacetate (121mg, 0.79mmol) was added to the reaction solution, and the reaction solution was stirred at room temperature for 2 hours. After the reaction was complete by LCMS, the reaction mixture was poured into saturated aqueous ammonium chloride (20 mL). The mixture was extracted with ethyl acetate (30mL X4), the organic phase was dried over anhydrous sodium sulfate and filtered, 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 give the desired product D28 (366 mg, yield: 87%, beige solid).
LCMS:Rt:0.77min；MS m/z(ESI):532.4[M+H]。
Intermediate preparation example 29: preparation of intermediate D29
Figure GDA0003667177710000261
Compound D23 (300mg, 1.5 mmol) and methylboronic acid (448.9mg, 7.5 mmol) were dissolved in dioxane (10 mL) and water (2 mL) under nitrogen, and Pd (dppf) Cl was added to the system 2 (110mg, 0.15mmol) and potassium phosphate (955mg, 4.5mmol); the reaction system is heated to 90 ℃ after being protected by nitrogen and stirred to react for 16 hours under the condition. After the reaction is finished, filtering the reaction solution, washing a filter cake by using ethyl acetate (20 mL), combining organic phases, drying and filtering, and concentrating the filtrate under reduced pressure; the residue was purified by column chromatography (elution gradient: PE: etOAc =10: 1-5:1) to give the objective product D29 (220 mg, yield: 80.3%, light yellow solid).
1 H NMR(400MHz,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
Figure GDA0003667177710000262
Compound D22 (120mg, 0.51mmol), cyclopropylboronic acid (263mg, 3.06mmol) was dissolved in 1,4-dioxane (18 mL) and 2mL K was added dropwise 2 CO 3 (211mg, 1.53mmol) of the solution, purging with Ar 3 times and adding Pd (dppf) Cl 2 (38mg, 0.051mmol), the mixture was fully aerated for 3 times, and heated to 80 ℃ for reaction overnight. LCMS to detect the completion of the reaction, cooling the reaction solution to room temperature, pouring into a separating funnel, adding water (50 mL) and ETOAC (20 mLx 3) for extraction, combining organic phases, washing with saturated NaCl solution (50 mL), and passing through anhydrous Na 2 SO 4 After drying, filtration, concentration of the filtrate to dryness and purification of the crude product on TLC prep. (PE: ETOAC = 2:1) gave the desired intermediate D30 (120 mg, yield: 100%, yellow oily liquid).
LCMS:Rt:1.553min；MS m/z(ESI):238.4[M+H]。
Intermediate preparation example 31: preparation of intermediate D31
Figure GDA0003667177710000263
Compound D30 (120mg, 0.51mmol) was dissolved in MeOH (20 mL) at room temperature, followed by the addition of Pd/C (20 mg); after the addition, the reaction system was replaced with hydrogen for 3 times, and then the system was stirred at room temperature for 2 hours. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd/C, and the filtrate was concentrated under reduced pressure to give the objective crude product D31 (80 mg, yield: 100%, yellow oily liquid). The crude product was used directly in the next reaction. LCMS: rt 0.542min; MS m/z (ESI) 208.3[ 2 ] M + H ].
Intermediate preparation example 32: preparation of intermediate D32
Figure GDA0003667177710000271
The compound (507mg, 3.0mmol), 5-fluoro-2,4-dichloropyrimidine (501mg, 3.0mmol) and NaHCO were added at room temperature 3 (756mg, 9.0mmol) in EtOH/H 2 O (8:1, 45 mL), and then the reaction was heated to 80 ℃ for overnight reaction. After the reaction was completed, the reaction solution was cooled to room temperature and diluted with water (100 mL), the mixture was extracted with ethyl acetate (50mL X3), the organic phases were combined and washed with a saturated aqueous solution of sodium chloride (100 mL), and then dried and filtered with anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure, and the residue was purified by prep-HPLC (eluent gradient:
Figure GDA0003667177710000272
) Purification gave the objective intermediate D32 (36 mg, yield: 4%, white solid).
LCMS:Rt:1.453min；MS m/z(ESI):300.3[M+H]。
Intermediate preparation 33: preparation of intermediate D33
Figure GDA0003667177710000273
The compound 5-bromo-2,4-dichloropyrimidine (5 g, 22.02mmol), the compound 2- (aminophenyl) dimethylphosphine oxide (3.7g, 22.02mmol) and NaHCO are reacted at room temperature 3 (5.5g, 66.06mmol) was dissolved in EtOH (50 mL) and water (10 mL). The reaction was heated to 100 ℃ and stirred for 10 hours, and the reaction was determined to be complete by LCMS. The reaction solution was cooled to room temperature and then diluted with water (50 mL), the mixture was extracted with dichloromethane (50 mL × 5), the organic phase was dried over sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent gradient PE/EtOAc =100:1 to 1).
LCMS:Rt:1.23min；MS m/z(ESI):362.1[M+H]。
Intermediate preparation example 34: preparation of intermediate D34
Figure GDA0003667177710000281
The compound (2-aminophenyl) dimethylphosphine oxide (450mg, 2.5 mmol), the compound 2,4-dichloro-5-methoxypyrimidine (505mg, 3.0 mmol) and DIETOAC (650mg, 5.0 mmol) were dissolved in DMF (10 mL) at room temperature, and the reaction mixture was stirred at 140 ℃ for 12 hours. After the reaction had been completed by LCMS, the reaction mixture was concentrated under reduced pressure and the residue was purified by preparative TLC (eluent gradient ETOAC/MeOH = 10.
LCMS:Rt:1.28min；MS m/z(ESI):312.2[M+H]。
Intermediate preparation example 35: preparation of intermediate D35
Figure GDA0003667177710000282
The compound 2,4-dichloro-5-methylpyrimidine (500mg, 3.07mmol) and the compound (2-aminophenyl) dimethylphosphine oxide (466mg, 2.76mmol) were dissolved in DMF (5 mL) at room temperature, followed by the addition of DIEA (1.18mg, 9.21mmol). The reaction solution was heated to 100 ℃ and stirred to react for 10 hours. LCMS to check the reaction was complete, the reaction was cooled to rt and diluted with water (20 mL), the mixture was extracted with dichloromethane (20 mL × 3), the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient PE/EtOAc =1, r f = 0.2) to obtain the target compound D35 (200 mg, yield: 22% white solid).
LCMS:Rt:1.20min；MS m/z(ESI):296.1[M+H]。
Intermediate preparation 36: preparation of intermediate D36
Figure GDA0003667177710000283
Compound E7 (390mg, 0.79mmol) and DIEA (308mg, 2.36mmol) were dissolved in DMF (15 mL) at room temperature, methyl bromoacetate (121mg, 0.79mmol) was added to the reaction solution, and the reaction solution was stirred at room temperature for 2 hours. After the reaction was complete by LCMS, the reaction mixture was poured into saturated aqueous ammonium chloride (20 mL). The mixture was extracted with ethyl acetate (30mL X4), the organic phase was dried over anhydrous sodium sulfate and filtered, 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 give the desired product D36 (366 mg, yield: 87%, beige solid).
LCMS:Rt:0.77min；MS m/z(ESI):532.4[M+H]。
Intermediate preparation example 37: preparation of intermediate D37
Figure GDA0003667177710000291
Compound D36 (106mg, 0.2mmol) was dissolved in MeOH/THF/H2O (4 mL/4mL/1 mL) at room temperature and LiOH (25mg, 0.6 mmol) was added, and the reaction was stirred at room temperature for 3 hours. After the reaction was completed by LCMS detection, the pH of the reaction mixture was adjusted to pH 3, and then the reaction mixture was directly concentrated under reduced pressure to obtain crude desired product D37 (104 mg, yield: 100%, white solid).
LCMS:Rt:0.29min；MS m/z(ESI):518.0[M+H]。
Intermediate preparation 38 and intermediate preparation 39: preparation of intermediates D38 and D39
Figure GDA0003667177710000292
The compound 2,4-dichloro-5-trifluoromethylpyrimidine (2.2 g, 10mmol), the compound (2-aminophenyl) dimethylphosphine oxide (2.0g, 12mmol) and DIEA (4.0g, 30mmol) were dissolved in DMF (25 mL) at room temperature, and the reaction mixture was stirred at room temperature for 16 hours. LCMS detected disappearance of starting material with product formation of both isomers. The reaction solution was concentrated under reduced pressure, and the resulting product (crude product: 8.0 g) was isolated by the following column chromatography conditions:
first batch: 4.0g of crude product, 330g of reverse phase preparative column, eluent gradient: acetonitrile/water = 0-100%, time =110 minutes; peak 1: 28-38 minutes; peak 2: 42-52 minutes.
And (2) second batch: 4.0g of crude product, 330g of reverse phase preparative column, eluent gradient: acetonitrile/water = 0-100%, time =130 minutes; peak 1: 35-41 minutes, peak 2: 50-55 minutes.
The two batches of Peak1 were combined and concentrated under reduced pressure to give product D38 (1.8 g, yield: 69%, yellow solid)
LCMS:Rt:1.099min；MS m/z(ESI):350.1[M+H]
1 H NMR(400MHz,MeOD)δ：11.1(s,1H),8.67(s,1H),8.21(m,1H),7.68(m,2H),7.32(t,J＝8.0Hz,1H),1.80(s,3H),1.76(s,3H)。
The two batches of Peak 2 were combined and concentrated under reduced pressure to give product D39 (0.7 g, yield: 69%, yellow solid)
LCMS:Rt:1.358min；MS m/z(ESI):350.1[M+H]，
1 H NMR(400MHz,MeOD)δ：11.9(s,1H),8.85(s,1H),8.48(m,1H),7.68(m,2H),7.24(t,J＝8.0Hz,1H),1.83(s,3H),1.79(s,3H)。
Intermediate preparation example 40: preparation of intermediate D40
Figure GDA0003667177710000301
Compound D38 (350mg, 1.0 mmol) and intermediate D5 (420mg, 1.5 mmol) were dissolved in EtOH (30 mL) at room temperature, and NH was added to the reaction system 4 After Cl (535mg, 10.0mmol), the reaction mixture was heated to 80 ℃ and stirred for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with ethanol (10mL X2), the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient DCM: meOH =20, white solid) to obtain the objective intermediate D40 (420 mg, yield: 70.8%.
LCMS:Rt:1.593min；MS m/z(ESI):594.5[M+H]。
Intermediate preparation example 41: preparation of intermediate D41
Figure GDA0003667177710000302
Compound D39 (350mg, 1.0 mmol) and intermediate D5 (420mg, 1.5 mmol) were dissolved in EtOH (30 mL) at room temperature, and NH was added to the reaction system 4 After Cl (535mg, 10.0mmol), the reaction mixture was heated to 80 ℃ and stirred for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with ethanol (10mL X2), the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient DCM: meOH = 20) to obtain the objective intermediate D41 (450 mg, yield: 75.8%, white solid).
LCMS:Rt:1.723min；MS m/z(ESI):594.6[M+H]。
Intermediate preparation example 42: preparation of intermediate D42
Figure GDA0003667177710000303
Compound D22 (200mg, 0.86mmol) and vinylboronic acid pinacol ester (800mg, 5.18mmol) were dissolved in 1,4-dioxane (18 mL) at room temperature, and then K was added dropwise 2 CO 3 (358mg, 2.59mmol) in water (2 mL), the reaction system was replaced with argon 3 times, and Pd (dppf) Cl was added 2 (32mg, 0.043 mmol), and the system was replaced 3 times, and then heated to 80 ℃ to react overnight. The reaction was completed by LCMS, the reaction was cooled to room temperature and diluted with water (50 mL), the mixture was extracted with ethyl acetate (20mL X3), the organic phases were combined and washed with a saturated aqueous sodium chloride solution (50 mL), and then dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient: PE: ETOAC = 2:1) to give the objective intermediate D42 (170 mg, yield: 88.2%, yellow oily liquid).
LCMS:Rt:1.502min；MS m/z(ESI):224.3[M+H]。
Intermediate preparation 43: intermediate (II)Preparation of form D43
Figure GDA0003667177710000311
Pd/C (30 mg) was added to a solution of compound D42 (170mg, 0.73mmol) in MeOH (20 mL) at room temperature, the reaction was replaced with hydrogen 3 times, and the reaction was stirred at room temperature for 2h. The reaction was completed by LCMS, the reaction solution was filtered to remove Pd/C, and the filtrate was concentrated under reduced pressure to give the objective crude product D43 (150 mg, yield: 100%, yellow oily liquid). The crude product was used directly in the next reaction.
LCMS:Rt:0.303min；MS m/z(ESI):196.4[M+H]。
Intermediate preparation example 44: preparation of intermediate D44
Figure GDA0003667177710000312
The compound 4-nitropyrazole (2.0g, 17.7mmol), the compound 1,4-dioxaspiro [4.5 ]/can-8-yl methanesulfonate (4.17g, 17.7mmol) and potassium carbonate (4.9g, 35.4mmol) were added to acetonitrile (50 mL) at room temperature, and the reaction was heated to 80 ℃ under argon atmosphere and stirred overnight. After completion of the reaction, 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 separation of the two phases, the aqueous phase was extracted with ethyl acetate (20mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient PE/etaac = 10/1) to obtain the objective compound D44 (3 g, yield: 67%, white solid).
LCMS:Rt:1.198min；MS m/z(ESI):254[M+H]。
Intermediate preparation example 45: preparation of intermediate D45
Figure GDA0003667177710000313
Under the protection of argon, the THF (30 mL) solution of the compound D44 (2g, 8.66mmol) is cooled to-78 ℃, and then LiHMDS (13mL, 1M, 13mmol) is slowly added into the system dropwise; after the completion of the addition, the reaction mixture was stirred at this temperature for 30 minutes, and then a solution of hexachloroethane (4.1g, 17.3 mmol) in THF (14 mL) was added dropwise thereto, and the reaction mixture was stirred for 3 hours. After the reaction is completed, adding saturated ammonium chloride aqueous solution (100 mL) into the reaction solution to quench the reaction, and extracting the mixture by using ethyl acetate (30mL x 3); the organic phases were combined and dried over anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient PE/EtOAc =15/1 to 12/1) to give the objective compound D45 (1.8 g, yield: 78%, white solid).
LCMS:Rt:1.772min；MS m/z(ESI):288[M+H]。
Intermediate preparation example 46: preparation of intermediate D46
Figure GDA0003667177710000314
Compound D45 (1.5g, 5.23mmol), methylboronic acid (5.5g, 91.7mmol), pd (dppf) Cl at room temperature 2 (450mg, 0.61mmol) and sodium carbonate (5g, 47.2mmol) were added to a mixed solvent of 1,4-dioxane and water (20/4 mL), the reaction system was replaced three times with argon, and then heated to 90 ℃ to stir the reaction for 24 hours. After the reaction is completed, cooling the reaction solution to room temperature, then diluting the reaction solution with water (50 mL) and ethyl acetate (20 mL), and extracting the aqueous phase with ethyl acetate (20mL x 2) after two-phase separation; the organic phases were combined, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by thin layer chromatography (eluent gradient PE/ETOAC =20/1 to 12/1) to obtain the objective product D46 (1.2 g, yield: 97%, white solid).
LCMS:Rt:1.906min；MS m/z(ESI):268[M+H]。
Intermediate preparation 47: preparation of intermediate D47
Figure GDA0003667177710000321
Compound D46 (1.2 g,4.5 mmol) was dissolved in THF (20 mL) at room temperature, pd/C (content 10wt%,600 mg) was added, the reaction was replaced with hydrogen three times, and then the mixture was stirred at room temperature under a hydrogen atmosphere overnight. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the objective product D47 (1.1 g, yield: 98%, white solid).
LCMS:Rt:0.373min；MS m/z(ESI):238[M+H]。
Intermediate preparation example 48: preparation of intermediate D48
Figure GDA0003667177710000322
Compound D12 (900mg, 3.8mmol), compound D47 (640mg, 3.8mmol), and ammonium chloride (2.1g, 38mmol) were dissolved in ethanol (15 mL) at room temperature, and the reaction was heated to 80 ℃ under an argon atmosphere and reacted for 48 hours. After completion of the reaction, the reaction solution was cooled to room temperature, the mixture was filtered, the filtrate was concentrated under reduced pressure, the residue was dissolved in DCM/MeOH (20/1,100mL), washed with water (30mL X2), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the objective compound D48 (1.6 g, yield:80%, red solid).
LCMS:Rt:1.30min；MS m/z(ESI):517[M+H]。
Intermediate preparation example 49: preparation of intermediate D49
Figure GDA0003667177710000323
Dioxane hydrochloride (4M, 5 mL) solution was added to a methanol (5 mL) solution of Compound D48 (1.6 g) with ice bath protection, and the reaction mixture was stirred at room temperature overnight. After completion of the reaction was cooled to 0 ℃ and then neutralized to pH >7 by addition of saturated aqueous sodium bicarbonate solution, the resulting mixture was concentrated under reduced pressure, the residue was dissolved in DCM/MeOH (20/1, 100ml) and the mixture was washed with water (30mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent gradient DCM/MeOH = 40/1) to give the title 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
Figure GDA0003667177710000331
NaH (60mg, 1.5 mmol) was added portionwise to a solution of compound D7 (230mg, 1.0 mmol) in DMF (10 mL) at 0 ℃. After the addition was complete, the reaction was stirred at 0 ℃ for 30 minutes, followed by addition of MeOH (48mg, 1.5mmol). After the addition, the reaction mixture was stirred at 0 ℃ for 3 hours. After completion of the reaction, the reaction was quenched with a saturated aqueous ammonium chloride solution (50 mL), and the resulting mixture was extracted with ethyl acetate (20mL X3), and the organic phase was washed with water (50 mL) and a saturated brine (50mL X2), dried over anhydrous sodium sulfate, and filtered, and the filtrate was concentrated under reduced pressure. Crude intended product D50 (110 mg, yield: 48.4.1%, yellow liquid) was obtained and used directly in the next step.
LCMS:Rt:1.208min；MS m/z(ESI):228.4[M+H]。
Intermediate preparation example 51: preparation of intermediate D51
Figure GDA0003667177710000332
Compound D33 (200mg, 0.54mmol, compound cyclopropylboronic acid (52mg, 0.61mmol), K at room temperature 2 CO 3 (228mg, 1.662mmol) and Pd (dppf) Cl 2 (40mg, 0.054mmol) was added to 1,4 dioxane (5 mL) and H 2 O (0.1 mL). After the reaction solution was replaced with argon three times, the reaction solution was heated to 100 ℃ and stirred for 8 hours, and the reaction was detected to be complete by LCMS. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and the mixture was extracted with dichloromethane (20 mL. Times.3). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue passing thin layer colorChromatography (eluent gradient PE: etOAc =1:100, r f = 0.3) to obtain the target compound D51 (120 mg, yield: 68% white solid).
LCMS:Rt:1.28min；MS m/z(ESI):322.2[M+H]。
Intermediate preparation example 52: preparation of intermediate D52
Figure GDA0003667177710000333
MsCl (2.9g, 25mmol) was slowly added to a solution of tert-butyl 7-hydroxy-2-azaspiro [3.5] nonane-2-carboxylate (5.0 g, 21mmol) in dichloromethane (50 mL) at 0 deg.C, and the reaction mixture was stirred at room temperature for 2 hours. After the reaction is completed, adding saturated aqueous sodium bicarbonate (100 mL) into the reaction solution to quench the reaction; after separation of the two phases, the aqueous phase was extracted with ethyl acetate (50mL X3), the organic phases were combined and washed with water and saturated brine (100 mL), and then dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain the objective 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 53: preparation of intermediate D53
Figure GDA0003667177710000334
4-Nitropyrazole (4.1g, 12.8mmol), D52 (1.6g, 14.1mmol), and K were sequentially added to a three-necked flask at room temperature 2 CO 3 (2.7g, 19.2mmol) and DMF (50 mL). The reaction solution was stirred at 120 ℃ for 2 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, water (50 mL) and ethyl acetate (50 mL) were added to the reaction mixture, the two phases were separated, and the aqueous phase was extracted with ethyl acetate (50mL X3). Mixing the organic phases, drying with anhydrous sodium sulfate, filtering, and concentrating the filtrate under reduced pressureThe residue was purified by reverse phase chromatography (330 g column, eluent gradient: water/acetonitrile = 0-100%, time =100 min) to give the desired product D53 (2.4 g, 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 54: preparation of intermediate D54
Figure GDA0003667177710000341
LiHMDS (18ml, 18mmol) was slowly added to a solution of Compound D53 (2.4g, 7.1mmol) in THF (50 mL) at-60 ℃ and, after completion of the addition, the reaction mixture was stirred at this temperature for 30 minutes. Hexachloroethane (2.1g, 8.7 mmol) was added to the reaction in portions and the reaction was stirred for an additional 2 hours. The reaction was quenched by adding saturated aqueous ammonium chloride (50 mL) and the resulting mixture was extracted with ethyl acetate (20mL X3). The organic phase was washed with saturated brine (50 mL), then dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient: PE: ETOAC =10: 1-4:1) to give the objective 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
Figure GDA0003667177710000342
D54 (800mg, 2.2mmol), iron powder (300mg, 4.4mmol), ammonium chloride (600mg, 10mmol) and ethanol (10 mL) were added in this order to a single-necked flask at room temperature. The reaction mixture was stirred at 80 ℃ for 16 hours. After completion of the reaction, the reaction solution was filtered while it was hot, and the filtrate was concentrated under reduced pressure to give a crude target compound D55 (400 mg, yield: 50%, green solid) which was used directly in the next step.
LCMS:Rt:1.168min；MS m/z(ESI):285.3[M-56+H]。
Intermediate preparation 56: preparation of intermediate D56
Figure GDA0003667177710000343
A mixture of compound D55 (255mg, 0.84mmol), compound D12 (300mg, 0.90mmol) and ammonium chloride (510mg, 8.4 mmol) in ethanol (30 mL) was heated to 80 ℃ and the reaction was stirred for 72 hours. After completion of the reaction, the reaction solution was cooled to room temperature, 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 objective compound D56 (600 mg, yield: >100%, green solid) which was directly used in the next step.
LCMS:Rt:1.075min；MS m/z(ESI):620.4[M+H]。
Intermediate preparation example 57: preparation of intermediate D57
Figure GDA0003667177710000351
In a 100mL three-necked flask at room temperature were added the compound 2,4-dichloropyrimidine-5-carboxylic acid ethyl ester (2.0g, 9mmol), the compound 2-aminophenyl-dimethylphosphine oxide (1.9g, 10.8mmol), DIEA (3.5g, 27mmol) and DMF (20 mL), respectively. The mixture was heated to 25 ℃ and the reaction stirred for 16 hours. After completion of the reaction, the reaction mixture was poured into water (50 mL). The mixture was extracted with ethyl acetate (50mL X3), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (eluent gradient DCM/MeOH = 80.
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 58: preparation of intermediate D58
Figure GDA0003667177710000352
Compound D15 (611.7mg, 2mmol), meOH (5 mL) and Pd/C (61.2mg, 10%) were added sequentially at room temperature in a 100mL single vial. The reaction system was stirred at room temperature under a hydrogen atmosphere for 16 hours. The reaction was completed by LCMS detection, the reaction solution was filtered to remove Pd/C, and the filtrate was concentrated under reduced pressure to give crude desired 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 59: preparation of intermediate D59
Figure GDA0003667177710000353
Compound D50 (116mg, 0.51mmol) was dissolved in MeOH (20 mL) at room temperature, followed by the addition of Pd/C (20 mg); after the addition, the reaction system was replaced with hydrogen for 3 times, and then the system was stirred at room temperature for 2 hours. The reaction was completed by LCMS detection, pd/C was removed by filtration from the reaction mixture, and the filtrate was concentrated under reduced pressure to give the objective 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
Figure GDA0003667177710000354
D4 (1.0 g, 3.2mmol) was dissolved in a dioxane solution (6N, 10 mL) of hydrogen chloride gas at room temperature, and after the solid was completely dissolved, the reaction mixture was stirred at room temperature for 2 hours. Thin layer chromatography (petroleum ether: ethyl acetate = 1):1，R f = 0.2) showed disappearance of starting material and formation of product. The reaction solution was concentrated under reduced pressure to give the objective compound D60 (1.1 g, crude product, white solid). The crude product was used directly in the next reaction.
Intermediate preparation 61: preparation of intermediate D61
Figure GDA0003667177710000361
D60 (500mg, 2.36mmol) and 1,2-dichloroethane (10 mL) were added to a 100mL single-neck bottle at room temperature followed by aqueous formaldehyde (3 mL); after the mixture was stirred at room temperature for 1 hour, sodium borohydride triacetate (995mg, 4.72mmol) and 5 drops of acetic acid were added, and after the addition, the reaction mixture was allowed to react at room temperature for 10 hours. The reaction was complete by LCMS, the reaction was diluted with water (20 mL) and the mixture was extracted with dichloromethane (20 mL. Times.3); the organic phases are combined and dried over anhydrous sodium sulfate, the filtrate is concentrated under reduced pressure, and the residue is purified by preparative thin-layer chromatography (eluent gradient: dichloromethane/methanol =20 f = 0.2) to obtain a target compound D61 (200mg, yield:37.8%, white solid).
LCMS:Rt:0.32min；MS m/z(ESI):225.3[M+H]。
Intermediate preparation 62: preparation of intermediate D62
Figure GDA0003667177710000362
D61 (200mg, 0.89mmol) was dissolved in ethanol (5 mL) at room temperature and palladium on carbon catalyst (20 mg) was added and the reaction was heated to 60 ℃ under hydrogen atmosphere and stirred for 2h. The reaction was completed by LCMS detection, the mixture was cooled and filtered, and the filtrate was concentrated under reduced pressure to give the target compound D62 (150mg, yield 86%, white solid.
LCMS:Rt:0.32min；MS m/z(ESI):195.3[M+H]。
Intermediate preparation example 63: preparation of intermediate D63
Figure GDA0003667177710000363
(R) -tetrahydrofuran-3-ylmethanesulfonic acid methyl ester (2.0g, 12.03mmol), 4-nitropyrazole (1.5g, 13.24mmol) and N, N-dimethylformamide (20 mL) were sequentially added to a 50mL single-neck flask at room temperature, potassium carbonate (4.99g, 36.10mmol) was further added, and the reaction mixture was heated to 120 ℃ and stirred overnight. The reaction was checked for completion by LCMS, the reaction solution was cooled to room temperature, water (30 mL) and ethyl acetate (30 mL) were added, the two phases were separated, the organic phase was washed with water (50 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the desired crude product D63 (2.5 g, yield: 70%, white solid). The crude product was used directly in the next reaction.
LCMS:Rt:1.253min；MS m/z(ESI):184.4[M+H]。
Intermediate preparation 64: preparation of intermediate D64
Figure GDA0003667177710000364
D63 (300.0mg, 1.64mmol), 1,2-dichloroethane (425.6mg, 1.80mmol) and tetrahydrofuran (5 mL) were added to a 25mL three-necked flask, respectively, and the reaction system was cooled to-60 ℃ under a nitrogen atmosphere, followed by slow dropwise addition of lithium bis-trimethylsilyl amide (1.64mL, 1.64mmol). After the addition was complete, the reaction mixture was stirred at-60 ℃ for a further 2 hours. The reaction was quenched by LCMS, saturated aqueous ammonium chloride (25 mL) was added to the reaction mixture, the mixture was extracted with ethyl acetate (50 mL), and the organic phase was Na anhydrous 2 SO 4 Drying, filtering, and concentrating the filtrate 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 objective product D64 (110 mg, yield: 31.0%, white solid).
LCMS:Rt:2.463min；MS m/z(ESI):217.4[M+H]。
Intermediate preparation example 65: preparation of intermediate D65
Figure GDA0003667177710000371
D64 (110.0 mg, 0.51mmol), methylboronic acid (303.0 mg, 5.1mmol) were dissolved in dioxane/water (2 mL/0.5 mL) at room temperature, followed by addition of sodium carbonate (265.0 mg,2.5 mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (36.6 mg, 0.05mmol). The reaction was purged with nitrogen and heated to 100 ℃ with stirring overnight. After completion of the reaction, the reaction mixture was cooled and filtered, the filtrate was diluted with water (20 mL) and ethyl acetate (20 mL), 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 and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent gradient: petroleum ether: ethyl acetate =10, rf = 0.2) to obtain the objective product D65 (60.0 mg, yield: 60%, brown solid).
Intermediate preparation example 66: preparation of intermediate D66
Figure GDA0003667177710000372
D65 (60.0 mg,0.3 mmol) was dissolved in tetrahydrofuran (5 mL) at room temperature, followed by the addition of a palladium on carbon catalyst (30.0 mg, 50%). The reaction was stirred at room temperature for 2h under a hydrogen atmosphere. The reaction was completed by LCMS detection, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give the objective product D66 (50.0 mg, yield: 100%, yellow solid).
LCMS:Rt:0.273min；MS m/z(ESI):168.5[M+H]。
Intermediate preparation example 67: preparation of intermediate D67
Figure GDA0003667177710000373
At room temperature, adding sodium cyanide (212mg, 4.32mmol) into a solution of 5-chloro-4-nitro-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazole (500mg, 2.16mmol) in N, N-dimethylformamide (4.3 mL), heating the reaction system to 80 ℃ under the protection of argon, and stirring for reaction for 7 hours; after completion of the reaction, the reaction system was cooled to room temperature, then diluted with water (20 mL), and the mixture was extracted with ethyl acetate (10mL X3). The organic phases were combined and washed with water (30 mL), the organic solution was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give the crude title compound D67 (400mg, yield 60%, yellow solid).
LCMS:Rt:1.635min；MS m/z(ESI):461[M+H 2 O-H]。
Intermediate preparation example 68: preparation of intermediate D68
Figure GDA0003667177710000374
Palladium on carbon (specification 10wt%,100 mg) was added to a solution of D67 (200mg, 0.9 mmol) in tetrahydrofuran (5 mL) at room temperature, and the reaction was stirred at room temperature for 6 hours under a hydrogen atmosphere. After completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude target compound D68 (150mg, yield.
LCMS:Rt:1.254min；MS m/z(ESI):402[2M+NH 4 ]。
Intermediate preparation 69: preparation of intermediate D69
Figure GDA0003667177710000381
D33 (200mg, 0.55mmol), 4,4,5,5-tetramethyl-2- (isopropyl-1-en-2-yl) -1,3,2-dioxoborate (93mg, 0.55mmol), potassium carbonate (227mg, 1.65mmol) and 1,1' -bis diphenylphosphinoferrocene palladium dichloride (40mg, 0.05mmol) were dissolved in dioxane (10 mL) at room temperature. The reaction system is heated to 100 ℃ under the protection of nitrogen, and stirred for 8 hours. The reaction was checked for completion by LCMS, the reaction was cooled and diluted with water (20 mL), the mixture was extracted with dichloromethane (30 mL × 5), the organic phase was dried over sodium sulfate and 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 = 0.4) to give the title compound D69 (110 mg, yield: 62.5%, white solid).
LCMS:Rt:1.653min；MS m/z(ESI):322.2[M+H]。
Intermediate preparation example 70: preparation of intermediate D70
Figure GDA0003667177710000382
D69 (100mg, 0.3 mmol) was dissolved in ethyl acetate (10 mL) at room temperature, followed by addition of palladium on carbon catalyst (20 mg). The reaction system is stirred for 2 hours under the condition of hydrogen atmosphere and room temperature, and the reaction is detected to be complete 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 give the objective compound D70 (30 mg, yield: 30%, white solid).
LCMS:Rt:1.49min；MS m/z(ESI):324.2[M+H]。
Intermediate preparation 71: preparation of intermediate D71
Figure GDA0003667177710000383
D2 (640 mg, 2.16mmol) was dissolved in methanol (30 mL) at room temperature, and then a palladium on carbon catalyst (120 mg) was added to the solution, and the reaction was stirred at room temperature under a hydrogen atmosphere for 16 hours. The reaction was completed by LCMS detection, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give the objective crude product D71 (550 mg, yield: 95.6%, yellow oily liquid). The crude product was used directly in the next reaction.
LCMS:Rt:0.933min；MS m/z(ESI):267.4[M+H]。
Intermediate preparation 72: preparation of intermediate D72
Figure GDA0003667177710000391
D71 (225mg, 0.834mmol) and D33 (150mg, 0.417mmol) were dissolved in ethanol (30 mL) at room temperature, and ammonium chloride (225mg, 4.2mmol) was added to the reaction; the reaction was then heated to 80 ℃ and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled and 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 preparative thin layer chromatography (eluent gradient: dichloromethane: methanol = 20).
LCMS:Rt:1.425min；MS m/z(ESI):592.4[M+3]。
Intermediate preparation 73: preparation of intermediate D73
Figure GDA0003667177710000392
D72 (55mg, 0.093 mmol) was dissolved in hydrogen chloride in dioxane (4M, 10 mL) at room temperature, and the reaction was carried out at room temperature for 2 hours. The reaction was completed by LCMS detection, and the reaction solution was concentrated under reduced pressure to give the hydrochloride of the objective intermediate D73 (50 mg, yield: 100%, yellow solid).
LCMS:Rt:0.717min；MS m/z(ESI):490.2[M+H]。
Intermediate preparation 74: preparation of intermediate D74
Figure GDA0003667177710000393
D3 (1.5g, 5 mmol), iron powder (1.4g, 25mmol) and ammonium chloride (2.16g, 40mmol) were added to a mixture of ethanol (28 mL) and water (7 mL) at room temperature, and the reaction was heated to 80 ℃ under argon and stirred for 1 hour. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain a target crude product D74 (1.3 g, yield: 96%), properties: a red solid.
LCMS:Rt:1.395min；MS m/z(ESI):245.2[M-56+H]。
Intermediate preparation example 75: preparation of intermediate D75
Figure GDA0003667177710000394
Figure GDA0003667177710000401
D74 (1.16g, 3.7 mmol), D12 (1g, 3.7 mmol) and ammonium chloride (951mg, 18.5 mmol) were dissolved in ethanol (30 mL) at room temperature, and the reaction was stirred at 80 ℃ for 12 hours. After completion of the reaction, the reaction mixture was poured into water (100 mL), the mixture was extracted with methylene chloride/methanol (10/1, 30 mLX) and the organic phase was dried over anhydrous sodium sulfate and filtered 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 (1.2 g, yield: 54%), properties: a white solid.
LCMS:Rt:1.539min；MS m/z(ESI):580.4[M+H]。
Intermediate preparation example 76: preparation of intermediate D76
Figure GDA0003667177710000402
D75 (220mg, 0.38mmol) and a 4M solution of hydrogen chloride dioxane (0.95mL, 3.8mmol) were dissolved in absolute ethanol (4 mL) at room temperature, and the reaction mixture was stirred at room temperature for 2 hours. The reaction was completed by LCMS detection, and the reaction mixture was concentrated under reduced pressure to give crude desired product D76 (182 mg, yield: 95%) as a pale yellow solid.
LCMS:Rt:1.62min；MS m/z(ESI):480.1[M+H]。
Intermediate preparation example 77: preparation of intermediate D77
Figure GDA0003667177710000403
7- (4-Nitro-1H-pyrazol-1-yl) -2-azaspiro [3.5] nonane (156mg, 0.66mmol), 1-bromo-2-methoxyethane (109mg, 0.79mmol) and potassium carbonate (96mg, 0.79mmol) were added to acetonitrile (5 mL) at room temperature, and then the reaction solution was heated to 80 ℃ and stirred for 16 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude target compound D77 (120 mg, yield: 91%), properties: yellow solid.
LCMS:Rt:0.716min；MS m/z(ESI):295.2[M+H]。
Intermediate preparation 78: preparation of intermediate D78
Figure GDA0003667177710000404
Wet palladium on carbon (12 mg) was added to a solution of D77 (120mg, 041mmol) in ethanol (5 mL) at room temperature, and the reaction was heated to 30 ℃ under a hydrogen atmosphere and stirred for 16 h. After completion of the reaction, the reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude target compound D78 (100 mg, yield: 93%) as a substance having the following properties: white solid
LCMS:Rt:0.312min；MS m/z(ESI):265.4[M+H]。
Intermediate preparation example 79: preparation of intermediate D79
Figure GDA0003667177710000411
Trifluoroacetic acid (2 mL) was added to a solution of D53 (300mg, 0.86mmol) in dichloromethane (8 mL) at room temperature, and the reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the objective compound D79 (312 mg, yield: 100%), properties: white solid, used directly in the next step.
LCMS:Rt:0.727min；MS m/z(ESI):237.2[M+H]。
Intermediate preparation example 80: preparation of intermediate D80
Figure GDA0003667177710000412
D79 Triethylamine (66.7 mg, 0.66mmol) was added to a solution of (156mg, 0.66mmol) and aqueous formaldehyde (198mg, 37% aqueous solution, 10.0 mmol) in methanol (5 mL) at room temperature, and after 5 hours of reaction with stirring, sodium triacetoxyborohydride (699.6 mg,3.3 mmol) was added and the reaction mixture was further reacted with stirring at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by thin layer preparative chromatography (eluent gradient: dichloromethane/methanol = 10/1) to obtain the objective compound D80 (100 mg, yield: 60%), properties: white solid LCMS: rt 0.808min; MS m/z (ESI) 321.5[ 2 ] M + H ].
Intermediate preparation example 81: preparation of intermediate D81
Figure GDA0003667177710000413
At room temperature, wet palladium on carbon (8.0 mg) was added to a solution of D80 (80mg, 0.32mmol) in ethanol (5 mL), and the reaction was heated to 30 ℃ under a hydrogen atmosphere and stirred for 4 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the objective compound D81 (56 mg, yield: 80%), properties: yellow solid
LCMS:Rt:0.291min；MS m/z(ESI):221.2[M+H]。
Intermediate preparation 82: preparation of intermediate D82
Figure GDA0003667177710000414
D77 (1066 mg, 6.16mmol) and diisopropylethylamine (751mg, 6.16mmol) were dissolved in acetonitrile (10 mL) at room temperature, and the reaction was heated to 80 ℃ and stirred for 16 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to give a crude target compound D82 (560 mg, yield: 65%), properties: yellow solid.
LCMS:Rt:0.885min；MS m/z(ESI):283.3[M+H]。
Intermediate preparation example 83: preparation of intermediate D83
Figure GDA0003667177710000415
Wet palladium on carbon (56 mg) was added to a solution of D82 (560mg, 1.98mmol) in ethanol (10 mL) at room temperature, and the reaction was heated to 30 ℃ under a hydrogen atmosphere and stirred for 16 hours. After completion of the reaction, the reaction solution was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude target compound D83 (360 mg, yield: 93%) as a substance having the following properties: yellow solid.
LCMS:Rt:0.259min；MS m/z(ESI):253.1[M+H]。
Intermediate preparation example 84: preparation of intermediate D84
Figure GDA0003667177710000421
Tert-butyl 5-oxocyclopentylalkyl [ c ] pyrrolidine-2 (1H) -carboxylic acid ester (10g, 44.4mmol) was dissolved in ethanol (200 mL) at room temperature, and then cooled to 0 ℃ to slowly add sodium borohydride (5.1g, 133.2mmol) to the reaction system, and the reaction was allowed to react at room temperature overnight. The completion of the reaction was checked by TLC, water (100 mL) was slowly added dropwise to the reaction mixture, the compound was extracted with ethyl acetate (50mL X4), the organic phases were combined and washed with saturated brine (50mL X3), and then dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain crude target compound D84 (9 g, yield: 95%), and properties: a yellow liquid.
1 H NMR(400MHz,CDCl 3 )δ4.28-4.21(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
Figure GDA0003667177710000422
D84 (10.0g, 44.4 mmol) was dissolved in methylene chloride (200 mL) at room temperature, triethylamine (9.0g, 88.8 mmol) was added, the reaction solution was cooled to-5 ℃ and then methanesulfonyl chloride (10.2g, 88.8 mmol) was slowly added to the system, the temperature of the system was controlled to not more than 5 ℃, and after completion of the dropwise addition, the reaction was allowed to stir at room temperature overnight. LCMS to check reaction completion, add saturated sodium bicarbonate solution (100 mL), stir for 10 minutes, extract the mixture with dichloromethane (50mL X3), combine the organic phases, dry filter with anhydrous sodium sulfate, concentrate the filtrate under reduced pressure to give crude target product D85 (1.5 g, yield: 70%), property: yellow solid. The crude product was used directly in the next reaction.
1 H NMR(400MHz,CDCl 3 ):δ5.17-5.04(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
Figure GDA0003667177710000423
D85 (5.2g, 46mmol), 4-nitro-1H-pyrazole (14g, 46mmol) and potassium carbonate (12.7g, 92mmol) were added to N, N-dimethylformamide (100 mL) at room temperature, and the reaction mixture was heated to 120 ℃ and stirred overnight. LCMS to check the reaction was complete, water (100 mL) was added to the reaction solution, the mixture was extracted with ethyl acetate (50mL X3), the combined organic phases were washed with saturated brine (50mL X5), the organic phase was dried over anhydrous sodium sulfate and filtered, 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 objective compound D86 (6 g, yield: 40%), properties: a yellow oil.
LCMS:Rt:1.231min；MS m/z(ESI):267[M-55]。
Intermediate preparation example 87: preparation of intermediate D87
Figure GDA0003667177710000424
Figure GDA0003667177710000431
D86 (1.5g, 4.6mmol) was dissolved in methanol (10 mL) at room temperature, and a solution of 1,4-dioxane (10 mL, 4N) in hydrogen chloride was added and the reaction stirred at room temperature for 2 hours. LCMS (liquid crystal display system) detects that the reaction is complete, and the reaction solution is decompressed and concentrated to obtain a crude target compound D87 (1.1 g, yield: 100%), and the characters are as follows: a white solid. The crude product was used directly in the next reaction.
LCMS:Rt:0.599min；MS m/z(ESI):223[M+H]。
Intermediate preparation example 88: preparation of intermediate D88
Figure GDA0003667177710000432
D87 (1.1g, 4.9mmol) was dissolved in methanol (20 mL) at room temperature, and after adding formaldehyde (1.5g, 15mL, 37%) the reaction mixture was stirred at room temperature overnight. LCMS indicated complete reaction, sodium triacetoxyborohydride (3.2g, 15mmol) was added and the reaction was stirred at room temperature for an additional 15 minutes. LCMS showed the reaction was complete, the reaction 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) to give the target compound D88 (800 mg, yield 68%), properties: a yellow oil.
LCMS:Rt:0476min；MS m/z(ESI):236[M+H]。
Intermediate preparation example 89: preparation of intermediate D89
Figure GDA0003667177710000433
D88 (800mg, 3.4 mmol) was dissolved in ethanol (20 mL) at room temperature, a palladium on carbon catalyst (200 mg) was added, the system was replaced three times with hydrogen gas, and the reaction was stirred at room temperature under a hydrogen atmosphere for 3 hours. LCMS shows that the reaction is complete, the reaction solution is filtered, and the filtrate is concentrated under reduced pressure to obtain a crude target compound D89 (500 mg, yield: 66%), and the character: a yellow oil. The crude product was used directly in the next reaction.
LCMS:Rt:0.233min；MS m/z(ESI):207[M+H]。
Intermediate preparation example 90: preparation of intermediate D90
Figure GDA0003667177710000434
D124 (1.2 g,2.2 mmol) and Lawson's reagent (1.78g, 4.4 mmol) were dissolved in toluene (50 mL) at room temperature, and the reaction was stirred at 120 ℃ for 3 hours. LCMS detection of product formation, reaction solution was filtered, filtrate was concentrated under reduced pressure, and residue was purified by silica gel column chromatography (eluent gradient: methanol: dichloromethane =0 to 10%) to obtain a target product D90 (350 mg, yield: 30%), character: light yellow solid. LCMS: rt 1.84min; MS m/z (ESI) 562.5[ 2 ] M + H ].
Intermediate preparation 91: preparation of intermediate D91
Figure GDA0003667177710000441
D90 (350mg, 0.62mmol) was dissolved in a 4M solution of hydrogen chloride dioxane (1.55mL, 6.2 mmol) at room temperature, and the reaction solution was stirred at room temperature for 2 hours. After completion of the reaction was detected by LCMS, the reaction mixture was concentrated under reduced pressure to obtain crude desired product D91 (310 mg, yield: 98%), properties: light yellow solid.
LCMS:Rt:1.15min；MS m/z(ESI):462.3[M+H]。
Intermediate preparation example 92: preparation of intermediate D92
Figure GDA0003667177710000442
At room temperature, tert-butyl 6-hydroxy-2-azaspiro [3.3] heptane-2-carboxylate (3.0g, 14.1mmol) was dissolved in 1,2-dichloromethane (30 mL), triethylamine (2.9g, 28.2mmol) was added, the reaction solution was cooled to 0 ℃ and methanesulfonyl chloride (1.6g, 14.1mmol) was slowly added to the system; the reaction was warmed to room temperature and stirred overnight. The reaction was completed by TLC, water (100 mL) was added to the reaction solution and stirred for 15 minutes, the mixture was extracted with dichloromethane (50mL X4), the organic phases were combined and washed with saturated brine (50 mLX), then dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain crude target compound D92 (2.8 g, yield: 70%) as a substance: yellow solid. The crude product was used directly in the next reaction.
LCMS:MS m/z(ESI):N/A。
Intermediate preparation example 93: preparation of intermediate D93
Figure GDA0003667177710000443
D92 (3.5g, 12mmol), 4-nitro-1H-pyrazole (1.4g, 12mmol) and potassium carbonate (3.3g, 24mmol) were added to N, N-dimethylformamide (50 mL) at room temperature, and the reaction mixture was heated to 120 ℃ and reacted overnight. Upon completion of the reaction by LCMS, water (100 mL) was added to the reaction solution, the mixture was extracted with ethyl acetate (50mL X3), the organic phases were combined and washed with saturated brine (50mL X5), and then dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain crude target compound D93 (2.0 g, yield: 70%) as a substance: yellow solid. The crude product was used directly in the next reaction.
LCMS:Rt:1.585min；MS m/z(ESI):309[M+H]。
Intermediate preparation 94: preparation of intermediate D94
Figure GDA0003667177710000444
Tert-butyl 6- (4-nitro-1H-pyrazol-1-yl) -2-azaspiro [3.3] heptane-2-carboxylate (500g, 1.6 mmol) was dissolved in methanol (10 mL) at room temperature, a solution of 1,4-dioxane (10mL, 4N) in hydrochloric acid gas was added, and the reaction was stirred at room temperature for 2 hours. LCMS (liquid Crystal display Mass Spectrometry) detects that the reaction is complete, and the reaction solution is decompressed and concentrated to obtain a crude target compound D94 (430 mg, yield: 100%), and the characters are as follows: a white solid. The crude product was used directly in the next reaction.
LCMS:Rt:0.353min；MS m/z(ESI):209[M+H]。
Intermediate preparation example 95: preparation of intermediate D95
Figure GDA0003667177710000451
D94 (430mg, 2.1 mmol) was dissolved in methanol (20 mL) at room temperature, and formaldehyde (511g, 6.3mL, 37%) was added and stirred at room temperature overnight. After the reaction was complete by LCMS, sodium triacetoxyborohydride (1.4 g,6.3 mmol) was added and the reaction was stirred for an additional 15 minutes at room temperature. LCMS showed completion of the reaction, 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) to obtain the target compound D95 (250 mg, yield: 55%), property: a yellow oil.
LCMS:Rt:0.463min；MS m/z(ESI):223[M+H]。
Intermediate preparation 96: preparation of intermediate D96
Figure GDA0003667177710000452
D95 (200mg, 0.9 mmol) was dissolved in ethanol (10 mL) at room temperature, and a palladium on carbon catalyst (100 mg) was added thereto, and the system was replaced with hydrogen three times and then stirred at room temperature under a hydrogen atmosphere for reaction for 3 hours. LCMS shows the reaction is complete, the reaction is filtered and the filtrate is concentrated under reduced pressure to give crude target compound D96 (150 mg, yield: 80%). The characteristics are as follows: a yellow oil. The crude product was used directly in the next reaction.
LCMS:Rt:0312min；MS m/z(ESI):193[M+H]。
Intermediate preparation example 97: preparation of intermediate D97
Figure GDA0003667177710000453
3- (benzyloxy) cyclobutanone (3.52g, 20.0mmol) and 1-methylpiperazine (2.20g, 22.0mmol) were dissolved in 1,2-dichloroethane (300 mL) at room temperature, and then sodium borohydride triacetate (8.46g, 40.0mmol) was added in portions; the reaction solution was stirred at room temperature for 3 hours. The reaction was completed by LCMS detection, and a saturated aqueous solution of sodium hydrogencarbonate (100 mL) was added to the system, and the compound was extracted with ethyl acetate (50 mLX), the organic phases were combined and washed with saturated brine (100 mL), followed by drying and filtration over anhydrous sodium sulfate, 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 objective product D97 (4.2 g, yield: 81%) as a colorless oil.
LCMS:Rt:1.75min；MS m/z(ESI):261.3[M+H]。
Intermediate preparation 98: preparation of intermediate D98
Figure GDA0003667177710000454
D97 (2.0g, 7.7mmol) was dissolved in trifluoroacetic acid (9 mL) at room temperature, and the reaction mixture was stirred at 80 ℃ for 12 hours. The reaction was completed by LCMS and the reaction mixture was concentrated under reduced pressure to give crude desired product D98 (1.31 g) as a pale yellow liquid.
LCMS:Rt:0.31min；MS m/z(ESI):171.4[M+H]。
Intermediate preparation example 99: preparation of intermediate D99
Figure GDA0003667177710000461
D98 (1.31g, 7.7 mmol) was dissolved in dichloromethane (50 mL) under ice-bath and triethylamine (3.85g, 38.5 mmol) was added followed by methanesulfonyl chloride (0.9mL, 11.55mmol). The reaction mixture was stirred at room temperature for 12 hours. To the system was added a saturated aqueous sodium bicarbonate solution (100 mL), the compound was extracted with ethyl acetate (20mL. Multidot.3), the combined organic phases were washed with a saturated brine (100 mL), and then dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude target product D99 (1.9 g) as a pale yellow liquid.
LCMS:Rt:0.43min；MS m/z(ESI):249.3[M+H]。
Intermediate preparation example 100: preparation of intermediate D100
Figure GDA0003667177710000462
D99 (1.9g, 7.7mmol), 4-nitro-1H-pyrazole (1.31g, 11.55mmol) and potassium carbonate (3.20g, 23.1mmol) were added to N, N-dimethylformamide (20 mL) at room temperature, and the reaction mixture was stirred at 120 ℃ for 12 hours. The product formation was checked by LCMS, the reaction was diluted with water (50 mL), the compound was extracted with ethyl acetate (40 mLX 2), the organic phases were combined and washed with saturated brine (100mL X2), then dried over anhydrous sodium sulfate and 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 give the desired product D100 (120 mg, three-step yield: 6%) as a pale yellow solid.
LCMS:Rt:0.43min；MS m/z(ESI):266.4[M+H]。
Intermediate preparation example 101: preparation of intermediate D101
Figure GDA0003667177710000463
D100 (120mg, 0.45mmol) and a palladium on carbon catalyst (60 mg) were added to absolute ethanol/tetrahydrofuran (10 mL/1 mL) at room temperature, the reaction solution was replaced with hydrogen gas three times, and then the reaction was stirred at room temperature for 1 hour. The reaction was completed by LCMS detection, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give crude desired product D101 (106 mg, yield: 98%) as a pale yellow solid.
LCMS:Rt:0.43min；MS m/z(ESI):236.3[M+H]。
Intermediate preparation 102: preparation of intermediate D102
Figure GDA0003667177710000464
D18 (153mg, 0.63mmol) and D12 (200mg, 0.63mmol) were dissolved in isopropanol (10 mL), and trifluoroacetic acid (1 mL) was added to react the reaction mixture at 100 ℃ for 10 hours. The reaction was completed by LCMS detection, 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 objective compound D102 (190 mg, yield: 73.9%), property: yellow solid.
LCMS:Rt:1.261min；(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
Figure GDA0003667177710000471
D102 (100mg, 0.24mmol) was dissolved in toluene (10 mL), thionyl chloride (0.5 mL) was added, and the reaction solution was heated to 70 ℃ for reaction for 4 hours. The reaction was completed by LCMS, and the reaction solution was concentrated under reduced pressure, and the residue was diluted with water (20 mL), followed by extraction with dichloromethane (20 mL × 3), the combined organic phases were dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain the objective compound D103 (80 mg, yield: 76.9%), properties: brown solid.
LCMS:Rt:1.421min；(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
Figure GDA0003667177710000472
D71 (500mg, 0.85mmol) and Lawson's reagent (688mg, 1.70mmol) were dissolved in toluene (10 mL) at room temperature, and the reaction mixture was stirred at 120 ℃ for 3 hours. LCMS detection of product formation, reaction solution was filtered, filtrate was concentrated under reduced pressure, and residue was purified by silica gel column chromatography (eluent gradient methanol/dichloromethane = 0-10%) to obtain the objective product D104 (330 mg, yield: 50%) as a pale yellow solid. LCMS: rt 1.85min; MS m/z (ESI) 608.2[ 2 ], M + H ].
Intermediate preparation 105: preparation of intermediate D105
Figure GDA0003667177710000473
D104 (330mg, 0.54mmol) and trifluoroacetic acid (2.0 mL) were dissolved in dichloromethane (5 mL) at room temperature, and the reaction was stirred at room temperature for 2 hours. The reaction was completed by LCMS, and the reaction mixture was concentrated under reduced pressure to obtain crude desired product D105 (210 mg, yield: 76%) as a pale yellow solid.
LCMS:Rt:1.16min；MS m/z(ESI):508.2[M+H]。
Intermediate preparation 106: preparation of intermediate D106
Figure GDA0003667177710000481
Tert-butyl 3-hydroxypiperidine-1-carboxylate (10g, 49.7mmol) and triethylamine (15g, 149mmol) were added to dichloromethane (150 mL) at room temperature. The mixture was replaced with argon for 3 times, cooled to-20 ℃ and then methanesulfonyl chloride (11.38g, 99.4 mmol) was slowly added dropwise to the system, after which the reaction was stirred at room temperature for 4 hours. After completion of the reaction, the reaction mixture was poured into a saturated aqueous sodium bicarbonate solution (300 mL), the mixture was extracted with ethyl acetate (100mL X3), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude target product D106 (12 g, yield: 87%), properties: a red liquid.
LCMS:Rt:1.615min；MS m/z(ESI):224.1[M-56+H]。
Intermediate preparation 107: preparation of intermediate D107
Figure GDA0003667177710000482
D106 (12g, 42.96mmol), 4-nitro-1H-pyrazole (9.71g, 85.91mmol) and potassium carbonate (17.81g, 128.88mmol) were dissolved in N' -dimethylformamide (160 mL) at room temperature, and the reaction was stirred at 110 ℃ for 12 hours. After completion of the reaction, the reaction mixture was poured into water (500 mL), the mixture was extracted with ethyl acetate (100mL X3), the organic phase was washed with a saturated aqueous sodium carbonate solution (300mL X8), dried and filtered with 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 =1, rf = 0.4) to obtain the objective product D107 (7.5 g, yield: 59%) for the property: a yellow liquid.
LCMS:Rt:1.772min；MS m/z(ESI):297.1[M+H]。
Intermediate preparation 108: preparation of intermediate D108
Figure GDA0003667177710000483
D107 (200mg, 0.67mmol) and a palladium on carbon catalyst (100 mg) were added to methanol (10 mL) at room temperature, and the reaction was substituted with hydrogen gas for 3 times and stirred at room temperature for 4 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the objective product D108 (170 mg, yield: 95%), properties: a yellow liquid.
LCMS:Rt:1.007min；MS m/z(ESI):267.2[M+H]。
Intermediate preparation 109: preparation of intermediate D109
Figure GDA0003667177710000491
D108 (170mg, 0.64mmol), D12 (202mg, 0.64mmol) and ammonium chloride (171mg, 3.19mmol) were dissolved in ethanol (8 mL) at room temperature and the reaction was stirred at 80 ℃ for 12 h. After completion of the reaction, the reaction mixture was poured into water (40 mL). The mixture was extracted with dichloromethane/methanol (10/1, 30mLX 3), the organic phase was dried over anhydrous sodium sulfate and filtered, 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 objective product D109 (110 mg, yield: 32%), properties: a white liquid.
LCMS:Rt:1.488min；MS m/z(ESI):546.2[M+H]。
Intermediate preparation example 110: preparation of intermediate D110
Figure GDA0003667177710000492
D109 (110mg, 0.2mmol) was dissolved in a mixture of hydrogen chloride in dioxane (4M, 0.6 mL) and methanol (1 mL) at room temperature, the reaction was stirred at room temperature for 1 hour, and the mixture was concentrated under reduced pressure to give crude target compound D110 (80 mg, yield: 89%), properties: a colorless liquid.
LCMS:Rt:0.804min；MS m/z(ESI):446.1[M+H]。
Intermediate preparation 111: preparation of intermediate D111
Figure GDA0003667177710000493
7- (4-Nitro-1H-pyrazol-1-yl) -2-azaspiro [3.5] nonane (100mg, 0.42mmol), 1,1-difluoro-2-iodoethane (160mg, 0.84mmol) and diisopropylethylamine (102mg, 0.84mmol) were dissolved in acetonitrile (5 mL) at room temperature, and then the reaction solution was heated to 80 ℃ and stirred for 16 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain crude target compound D111 (100 mg, yield: 91%), properties: black liquid
LCMS:Rt:0.823min；MS m/z(ESI):301.3[M+H]。
Intermediate preparation example 112: preparation of intermediate D112
Figure GDA0003667177710000501
Wet palladium on carbon (12 mg) was added to a solution of D111 (45mg, 015 mmol) in ethanol (5 mL) at room temperature, and the reaction was heated to 30 ℃ under a hydrogen atmosphere and stirred for 16 hours. After completion of the reaction, the reaction liquid was cooled to room temperature and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude target compound D112 (30 mg, yield: 75%), property: a black solid.
LCMS:Rt:1.028min；MS m/z(ESI):271.2[M+H]。
Intermediate preparation example 113: preparation of intermediate D113
Figure GDA0003667177710000502
Wet palladium-on-carbon (50 mg) was added to a solution of D53 (0.5g, 1.49mmol) in ethanol (30 mL) at room temperature, and the reaction was heated to 30 ℃ under a hydrogen atmosphere and stirred for 6 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude target compound D113 (1.8 g, yield: 100%), properties: a white solid.
LCMS:Rt:0.858min；MS m/z(ESI):307.1[M+H]。
Intermediate preparation 114: preparation of intermediate D114
Figure GDA0003667177710000503
D12 (1.65g, 5.2 mmol), D113 (1.6 g,5.2 mol) and ammonium chloride (2.76g, 5.2 mmol) were dissolved in ethanol (5 mL) at room temperature, and the reaction was heated to 80 ℃ and stirred overnight. After completion of the reaction, the reaction mixture 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: dichloromethane: methanol = 1:0-10.
LCMS:Rt:1.442min；MS m/z(ESI):586.5[M+H]。
Intermediate preparation 115: preparation of intermediate D115
Figure GDA0003667177710000504
Lawson's reagent (276mg, 0.683mmol) was added to a solution of D114 (400mg, 0.683mmol) in toluene (10 mL) at room temperature, and the reaction was stirred at 110 ℃ for 4 hours. After completion of the reaction, the reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography (eluent gradient: dichloromethane: methanol = 1:0-10) of the residue to obtain the objective intermediate D115 (250 mg, yield: 49%) as a white solid.
LCMS:Rt:1.845min；MS m/z(ESI):602.4[M+H]。
Intermediate preparation 116: preparation of intermediate D116
Figure GDA0003667177710000511
Trifluoroacetic acid (3 mL) was added to a solution of D115 (250mg, 0.41mmol) in dichloromethane (12 mL) at room temperature, and the reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a crude target compound D116 (180 mg, yield: 86%), properties: white solid, used directly in the next step.
LCMS:Rt:1.006min；MS m/z(ESI):502.3[M+H]。
Intermediate preparation example 117: preparation of intermediate D117
Figure GDA0003667177710000512
D84 (2.0 g, 8.47mmol) and a palladium/carbon catalyst (300 mg) were added to ethanol (30 mL) at room temperature, and the reaction solution was replaced three times with hydrogen gas, followed by stirring at 50 ℃ for 1 hour. LCMS detects that the reaction is complete, the reaction solution is filtered, and the filtrate is decompressed and concentrated to obtain the target compound D117 (1.6 g, crude product) with the character of yellow solid.
LCMS:Rt:0.49min；MS m/z(ESI):237.2[M+H]。
Intermediate preparation 118: preparation of intermediate D118
Figure GDA0003667177710000513
D117 (1.52mg, 7.91mmol), and D12 (2.5g, 7.91mmol) and ammonium chloride (2.09g, 39.55mmol) were dissolved in ethanol (30 mL). The reaction solution was stirred at 100 ℃ for 8 hours, LCMS was used to detect completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give the desired compound D118 (4.0 g, crude) as a yellow solid.
LCMS:Rt:1.665min；(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
Figure GDA0003667177710000521
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 detects that the reaction is complete, and the reaction solution is decompressed and concentrated to obtain crude target compound D119 (2.1 g, crude product) with yellow solid.
LCMS:Rt:1.165min；(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
Figure GDA0003667177710000522
Trifluoroacetic acid (2 mL) was added to a solution of D114 (100mg, 0.17mmol) in dichloromethane (8 mL) at room temperature, and the reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a crude target compound D120 (72 mg, yield: 86%), ZZ character: white solid, used directly in the next step.
LCMS:Rt:0.748min；MS m/z(ESI):486.2[M+H]。
Intermediate preparation 121: preparation of intermediate D121
Figure GDA0003667177710000523
D54 (200mg, 0.54mmol) was dissolved in trifluoroacetic acid (1 mL) and dichloromethane (5 mL), and the reaction mixture was reacted at room temperature for 2 hours. LCMS detects that the reaction is complete, and the reaction solution is decompressed and concentrated to obtain the target compound D121 (140 mg, crude product) with the character of yellow solid.
LCMS:Rt:0.94min；(0.1％FA-H 2 O):(0.1％FA-CAN)＝90:10；MS m/z(ESI):271.1[M+H]。
Intermediate preparation 122: preparation of intermediate D122
Figure GDA0003667177710000524
D121 (100mg, 0.37mmol), 1-bromo-2-methoxyethane (101mg, 0.74mmol) and N, N-diisopropylethylamine (143mg, 1.11mmol) were dissolved in acetonitrile (5 mL), and the reaction mixture was stirred at 70 ℃ for 10 hours. LCMS to check the reaction was complete, the reaction was concentrated under reduced pressure and the residue was chromatographed by thin layer preparative chromatography (eluent gradient: ethyl acetate: petroleum ether =1, r f = 0.6) to obtain the target compound D122 (95 mg, yield: 78.5%), character: white solid.
LCMS:Rt:1.08min；(0.1％FA-H 2 O):(0.1％FA-CAN)＝90:10；MS m/z(ESI):329.1[M+H]。
Intermediate preparation 123: preparation of intermediate D123
Figure GDA0003667177710000531
D122 (90mg, 0.27mmol), ammonium chloride (14mg, 0.27mmol) and iron powder (15mg, 0.27mmol) were added to water (2 mL) and ethanol (10 mL), and the reaction mixture was heated to 80 ℃ for 8 hours. LCMS detects that the reaction is complete, the reaction is filtered, and the filtrate is decompressed and concentrated to obtain a crude target compound D123 (70 mg, crude product) with brown solid.
LCMS:Rt:0.375min；(0.1％FA-H 2 O):(0.1％FA-CAN)＝90:10；MS m/z(ESI):299.4[M+H]。
Intermediate preparation 124: preparation of intermediate D124
Figure GDA0003667177710000532
D71 (1.7 g, 6.38mmol), D12 (2g, 6.38mmol) and ammonium chloride (3.4 g, 63.8mmol) were dissolved in ethanol (30 mL) at room temperature, and the reaction was stirred at 80 ℃ 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 mLX), the organic phase was dried over anhydrous sodium sulfate and filtered, 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 objective product D124 (2 g, yield: 57%), properties: yellow solid.
LCMS:Rt:1.562min；MS m/z(ESI):546.5[M+H]。
Intermediate preparation 125: preparation of intermediate D125
Figure GDA0003667177710000533
D124 (1g, 1.8mmol) was dissolved in a mixed solution of hydrogen chloride in dioxane (4M, 10mL) and methanol (10 mL) at room temperature, the reaction was stirred at room temperature for 1 hour, and then the mixture was concentrated under reduced pressure to obtain the objective compound D125 (800 mg, yield: 94%), property: a white solid.
LCMS:Rt:0.796min；MS m/z(ESI):446.3[M+H]。
Intermediate preparation 126: preparation of intermediate D126
Figure GDA0003667177710000541
2-chloro-N, N-dimethylethylamine hydrochloride (3g, 21.23mmol), 4-nitro-1H-pyrazole (2g, 17.69mmol), cesium carbonate (17.29g, 53mmol) and diisopropylethylamine (4.56g, 35.38mmol) were mixed in N' N-dimethylformamide (30 mL) at room temperature, and the reaction was stirred at 130 ℃ for 12 hours. After completion of the reaction, the reaction mixture was poured into water (300 mL); the mixture was extracted with dichloromethane/methanol (10/1, 100mL X3), the organic phase was dried over anhydrous sodium sulfate and filtered, 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 objective product D126 (2.2 g, yield: 68%), properties: yellow solid.
LCMS:Rt:0.394min；MS m/z(ESI):185.3[M+H]。
Intermediate preparation 127: preparation of intermediate D127
Figure GDA0003667177710000542
D126 (500mg, 2.71mmol) and a palladium on carbon catalyst (150 mg) were added to methanol (10 mL) at room temperature, the reaction system was replaced with hydrogen gas 3 times, and then the reaction solution was stirred under hydrogen at room temperature for 4 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the objective product D127 (400 mg, yield: 96%), properties: yellow solid.
LCMS:Rt:0.285min；MS m/z(ESI):155.2[M+H]。
Intermediate preparation 128: preparation of intermediate D128
Figure GDA0003667177710000543
D21 (400mg, 0.71mmol) and Lawson's reagent (578mg, 1.42mmol) were dissolved in toluene (10 mL) at room temperature, and the reaction mixture was stirred at 120 ℃ for 3 hours. 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 objective product D128 (200 mg, yield: 50%) as a pale yellow solid.
LCMS:Rt:2.25min；MS m/z(ESI):576.2[M+H]。
Intermediate preparation example 129: preparation of intermediate D129
Figure GDA0003667177710000544
Figure GDA0003667177710000551
D128 (100mg, 0.17mmol) was dissolved in a 4M solution of hydrogen chloride in dioxane (0.4 mL,1.7 mmol) at room temperature, and the reaction mixture was stirred at room temperature for 12 hours. The reaction was completed by LCMS detection, and the reaction mixture was concentrated under reduced pressure to give crude desired product D129 (81 mg, yield: 98%) as a pale yellow solid.
LCMS:Rt:2.10min；MS m/z(ESI):476.2[M+H]。
Intermediate preparation 130: preparation of intermediate D130
Figure GDA0003667177710000552
D118 (600mg, 1.0 mmol), lawson's test, was added at room temperatureThe reagent (808mg, 2.0mmol) was dissolved in toluene (30 mL), and the reaction mixture was heated to 110 ℃ and stirred for 4 hours. After completion of the reaction, the reaction mixture was poured into water (100 mL), the mixture was extracted with dichloromethane/methanol (10/1, 30 mLX) and the organic phase was extracted with anhydrous Na 2 SO 4 The filtrate was concentrated under reduced pressure, and the residue was 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%), properties: white solid.
LCMS:Rt:1.894min；MS m/z(ESI):588.5[M+H]。
Intermediate preparation 131: preparation of intermediate D131
Figure GDA0003667177710000553
D130 (100mg, 0.17mmol) was dissolved in a mixture of hydrogen chloride in dioxane (4M, 2mL) and methanol (1 mL) at room temperature, the reaction was stirred at room temperature for 1 hour, and the mixture was concentrated under reduced pressure to obtain the objective compound D131 (80 mg, yield: 95%) as a solid product: yellow solid.
LCMS:Rt:1.225min；MS m/z(ESI):488.1[M+H]。
Intermediate preparation 132: preparation of intermediate D132
Figure GDA0003667177710000554
Tert-butyl 9-oxo-3-azaspiro [5.5] undecane-3-carboxylate (500mg, 1.87mmol) was dissolved in methanol (10 mL) at room temperature, then sodium borohydride (171mg, 4.5 mmol) was added in portions, and the reaction solution was stirred at room temperature for 1 hour. The reaction was completed by LCMS, and saturated aqueous sodium bicarbonate (50 mL) was added to the system, followed by extraction with ethyl acetate (20mL. Multidot.3), the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give crude desired product D132 (500 mg, yield: 60%) as a colorless oil.
LCMS:Rt:1.75min；MS m/z(ESI):214.5[M-56+H]。
Intermediate preparation 133: preparation of intermediate D133
Figure GDA0003667177710000561
D132 (500mg, 1.87mmol) was dissolved in dichloromethane (20 mL) under ice-bath, triethylamine (0.77mL, 5.61mmol) was added, followed by methanesulfonyl chloride (0.29mL, 3.74mmol); the reaction mixture was stirred at room temperature for 12 hours. The reaction was quenched with saturated aqueous sodium bicarbonate (50 mL), extracted with dichloromethane (20 mLX), the combined organic phases were washed with saturated brine (50 mL), then dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent gradient: petroleum ether/ethyl acetate = 5:1) to give the objective product D133 (100 mg, yield: 16%) as a colorless oil.
LCMS:Rt:1.89min；MS m/z(ESI):292.4[M-56+H]。
Intermediate preparation 134: preparation of intermediate D134
Figure GDA0003667177710000562
D133 (90mg, 0.26mmol), 4-nitro-1H-pyrazole (58g, 0.52mmol) and potassium carbonate (108mg, 0.78mmol) were added to N, N-dimethylformamide (2 mL) at room temperature, and the reaction mixture was stirred at 110 ℃ for 12 hours. The product formation was checked by LCMS, the reaction was diluted with water (20 mL), the compound was extracted with ethyl acetate (40 mLX), the organic phases were combined and washed with saturated brine (100mL X2), then dried over anhydrous sodium sulfate filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by preparative thin layer chromatography (eluent gradient: methanol/dichloromethane = 1) to give the desired product D134 (50 mg, yield: 52%) as a white solid.
LCMS:Rt:2.04min；MS m/z(ESI):309.3[M-56+H]。
Intermediate preparation 135:preparation of intermediate D135
Figure GDA0003667177710000563
Tert-butyl 9- (4-nitro-1H-pyrazol-1-yl) -3-azaspiro [5.5] undecane-3-carboxylic acid ester (50mg, 0.14mmol) and a palladium on carbon catalyst (30 mg) were added to absolute ethanol (3 mL) at room temperature, and the reaction solution was replaced with hydrogen gas three times and stirred at room temperature for 2 hours. The reaction was completed by LCMS detection, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give crude target compound D135 (48 mg, yield: 98%) as a pale yellow solid. LCMS, rt is 1.16min; MS m/z (ESI) 335.2[ 2 ] M + H ].
Intermediate preparation 136: preparation of intermediate D136
Figure GDA0003667177710000571
D135 (48mg, 0.14mmol), D12 (44mg, 0.14mmol) and ammonium chloride (38mg, 0.70mmol) were dissolved in isopropanol (3 mL) at room temperature, and the reaction solution was stirred at 80 ℃ for 3 hours. The reaction was checked for completion by LCMS, the reaction was diluted with saturated aqueous sodium bicarbonate (10 mL) and then extracted with ethyl acetate (20 mLX), the combined organic phases were washed with saturated brine (50 mL), then dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by preparative thin layer chromatography (eluent gradient: methanol/dichloromethane = 1) to give the target product D136 (32 mg, yield: 37%) as a white solid.
LCMS:Rt:1.89min；MS m/z(ESI):614.2[M-56+H]。
Intermediate preparation example 137: preparation of intermediate D137
Figure GDA0003667177710000572
D136 (32mg, 0.05mmol) was dissolved in a 4M dioxane solution of hydrogen chloride (0.13mL, 0.5mmol) at room temperature, and the reaction mixture was stirred at room temperature for 3 hours. The reaction was completed by LCMS detection, and the reaction mixture was concentrated under reduced pressure to give crude desired product D137 (30 mg, yield: 98%) as a pale yellow solid.
LCMS:Rt:0.37min；MS m/z(ESI):514.2[M+H]。
Intermediate preparation 138: preparation of intermediate D138
Figure GDA0003667177710000573
3- (benzyloxy) cyclobutanone (5 g,28.4 mmol) and tert-butylpiperazine-1-carboxylate (5.28g, 28.4 mmol) were dissolved in 1,2-dichloroethane (500 mL) at room temperature, sodium borohydride triacetate (11.98g, 56.8mmol) was added in portions, and the reaction mixture 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 phases were dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (eluent gradient: petroleum ether/ethyl acetate =100:1 to 2:1) to obtain the objective compound D138 (8.25 g, yield: 83.9%) as a white solid.
LCMS:Rt:1.345min；(0.1％FA-H 2 O):(0.1％FA-ACN)＝90:10；MS m/z(ESI):347.5[M+H]。
Intermediate preparation 139: preparation of intermediate D139
Figure GDA0003667177710000574
D138 (6.0 g, 17.34mmol) was dissolved in trifluoroacetic acid (20 mL) at room temperature, and the reaction was stirred at 80 ℃ for 6 hours. The reaction was complete by LCMS, the reaction mixture was concentrated to give crude which was dissolved in dichloromethane (5 mL) and triethylamine (5.25g, 52.02mmol) and di-tert-butyl dicarbonate (7.56g, 34.68mmol) were added and 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 phases were combined and concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluent gradient: dichloromethane/methanol =100:1 to 10) to obtain the target compound D139 (3.9 g, yield: 88.04%) as a yellow oil.
LCMS:Rt:1.006min；(0.1％FA-H 2 O):(0.1％FA-ACN)＝90:10；MS m/z(ESI):257.5[M+H]。
Intermediate preparation 140: preparation of intermediate D140
Figure GDA0003667177710000581
D139 (3.0g, 11.71mmol) was dissolved in methylene chloride (50 mL) and triethylamine (3.5g, 35.13mmol) was added, followed by methanesulfonyl chloride (2.6mg, 23.43mmol); the reaction solution was stirred at room temperature for 10 hours. Water (50 mL) was added to the system, the mixture was extracted with dichloromethane (50 mL. Times.4), the combined organic phases were washed with a saturated aqueous solution of sodium hydrogencarbonate (50 mL. Times.3), and then dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give crude title compound D140 (4.0 g, crude) as a pale yellow liquid.
LCMS:Rt:1.145min；(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
Figure GDA0003667177710000582
D140 (1.5g, 4.49mmol), 4-nitro-1H-pyrazole (507mg, 4.49mmol) and potassium carbonate (1.85g, 4.49mmol) were added to N' -N-dimethylformamide (20 mL) at room temperature, and the reaction mixture was stirred at 110 ℃ for 10 hours. LCMS to check the reaction was complete, diluted with water (50 mL), the mixture was extracted with ethyl acetate (30 mL × 4), the combined organic phases were washed with saturated aqueous potassium carbonate (50mL × 3), then dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluent gradient: ethyl acetate/petroleum ether = 1:5-ethyl acetate/methanol = 10) to obtain the objective compound D141 (650 mg, yield: 43.33%) as a white solid.
1 H NMR(400MHz,MeOD)：δ8.63(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 142: preparation of intermediate D142
Figure GDA0003667177710000583
D141 (650mg, 1.834mmol) and a palladium/carbon catalyst (300 mg) were added to tetrahydrofuran (30 mL) at room temperature, and the reaction solution was replaced with hydrogen gas three times, followed by stirring at room temperature for 10 hours. The reaction was completed by LCMS detection, palladium/carbon was filtered off, and the filtrate was concentrated under reduced pressure to give the objective compound D142 (610 mg, crude product) as a pale yellow solid.
LCMS:Rt:0.303min；(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
Figure GDA0003667177710000591
D142 (509mg, 1.58mmol), D12 (500g, 1.58mmol) and ammonium chloride (334g, 6.32mmol) were dissolved in ethanol (5 mL); the reaction solution was stirred at 110 ℃ for 10 hours. LCMS detects completion of the reaction, the reaction solution is filtered, the filtrate is concentrated under reduced pressure, and the obtained crude product is purified by silica gel column chromatography (eluent gradient: dichloromethane/methanol =100:1 to 10.
LCMS:Rt:1.250min；(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
Figure GDA0003667177710000592
D143 (150mg, 0.25mmol) was dissolved in hydrogen chloride in dioxane (5 mL) and methanol (1 mL) and the reaction was stirred at room temperature for 1.5 h. LCMS detects that the reaction is complete, and the reaction solution is decompressed and concentrated to obtain the target compound D144 (130 mg, crude product) with the character of yellow solid.
LCMS:Rt:0.780min；(0.1％FA-H2O):(0.1％FA-ACN)＝90:10；MS m/z(ESI):501.2[M+H]。
Intermediate preparation 145: preparation of intermediate D145
Figure GDA0003667177710000593
D143 (200mg, 0.33mmol) and Lawson's reagent (271mg, 0.67mmol) were dissolved in toluene (10 mL) at room temperature, and the reaction was stirred at 110 ℃ for 4 hours. After completion of the reaction, the reaction mixture was poured into water (100 mL), extracted with dichloromethane/methanol (10/1, 30 mLX), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure; the residue was purified by silica gel column chromatography (eluent gradient: dichloromethane: methanol =20:1, rf = 0.6) to obtain the objective product D145 (120 mg, yield: 59%), properties: yellow solid.
LCMS:Rt:1.019min；MS m/z(ESI):517.1[M-100+H]。
Intermediate preparation 146: preparation of intermediate D146
Figure GDA0003667177710000601
D145 (120mg, 0.19mmol) was dissolved in a mixture of hydrogen chloride in dioxane (4M, 4mL) and methanol (1 mL) at room temperature, the reaction was stirred at room temperature for 1 hour, and the mixture was concentrated under reduced pressure to give crude target compound D146 (100 mg, yield 99%) as a solid product: yellow solid.
LCMS:Rt:2.118min；MS m/z(ESI):517.2[M+H]。
Example 1 and example 1'
Dimethyl (2- ((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) -5- (tris Fluoromethyl) pyrimidin-4-yl) amino) phenyl) phosphine oxide 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
Figure GDA0003667177710000602
A mixture of compounds D10 and D10' (140mg, 0.42mmol) and compound D9 (79mg, 0.44mmol) were dissolved in EtOH (10 mL), and after ammonium chloride (106.9mg, 2.0 mmol) was added to the reaction system, the reaction was stirred at 105 ℃ 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 purified by preparative high performance liquid chromatography (eluent gradient:
gradient conditions:
Figure GDA0003667177710000603
) Purification gave pure product E1' (trifluoroacetate, 14.5mg, yield: 7.3%) and product E1 (trifluoroacetate salt, 74.5mg, yield: 37.6%).
E1’:
HPLC：99.152％@214nm，99.42％@254nm
LCMS：MS m/z(ESI):495.3[M+H]
19 F NMR(400MHz,CD 3 OD)δ：-62.90,-63.28,-77.23.
1 H NMR(400MHz,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/z(ESI):495.3[M+H]
19 F NMR(400MHz,CD 3 OD)δ：-64.17,-77.37.
1 H NMR(400MHz,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- ((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) -5- (tris Fluoromethyl) pyrimidin-4-yl) amino) phenyl) phosphine oxide
Figure GDA0003667177710000611
Compound D11 (100mg, 0.6 mmol) and intermediate D12 (158mg, 0.5 mmol) were dissolved in isopropanol (20 mL) at room temperature, trifluoroacetic acid (1.14g, 10.0 mmol) was added to the reaction system, and the reaction mixture was heated to 120 ℃ and stirred for 3 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000612
) Purification gave the desired product E2 (trifluoroacetate salt, 29.1mg, yield: 13% white solid).
HPLC：99.83％@214nm，99.81％@254nm
LCMS：MS m/z(ESI):447.4[M+H]
19 F NMR (376.5 MHz, methanol-d) 4 )δ-74.71(s)
1 H NMR(400MHz,DMSO-d 6 )δ10.94(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＝11.2Hz,2H),1.89-1.75(m,10H).
Example 3
(2- ((5-chloro-1- (2-hydroxyethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) benzene Yl) dimethyl phosphine oxide (E3)
Figure GDA0003667177710000621
Compound D58 (165mg, 0.6 mmol) and intermediate D12 (158mg, 0.5 mmol) were dissolved in isopropanol (20 mL) at room temperature, trifluoroacetic acid (1.14g, 10.0 mmol) was added to the reaction system, and the reaction mixture was heated to 120 ℃ and stirred for reaction for 3 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000622
) Purification gave the desired product E3 (trifluoroacetate salt, 15.3mg, yield: 16%), property: a white solid.
HPLC：99.11％@214nm，98.96％@254nm
LCMS：MS m/z(ESI):441.3[M+H]
1 H NMR (400 MHz, 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＝5.6Hz,2H),3.92(t,J＝5.6Hz,2H),1.89(s,3H),1.85(s,3H).
19 F NMR (376.5 MHz, methanol-d) 4 )δ-77.25(s)
Example 4
(5-chloro-2- (2- (1- (2-hydroxyethyl) -1-hydropyrazol-4-amino) pyrimidin-4-amino) phenyl) dimethylphosphine oxide Article (A)
Figure GDA0003667177710000623
Compound D17 (100mg, 0.70mmol) and intermediate D12 (223.0mg, 0.7mmol) were dissolved in isopropanol (5 mL) at room temperature, and after trifluoroacetic acid (1596.0mg, 14.0mmol) was added to the reaction system, the reaction was stirred at 120 ℃ 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 purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000624
Figure GDA0003667177710000631
) Purification gave the desired product E4 (trifluoroacetate salt, 47.4mg, yield: 16%, white solid).
HPLC：99.84％@214nm，99.83％@254nm
LCMS：MS m/z(ESI):421.2[M+H]
1 H NMR(400MHz,DMSO-d 6 )δ10.98(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＝5.3Hz,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) Dimethyl phosphine oxide
Figure GDA0003667177710000632
After completion of Compound D18 (100mg, 0.4 mmol) and intermediate D12, the reaction mixture was stirred at 80 ℃ for 16 hours at room temperature. 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 purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000633
) Purification gave the desired product E5 (trifluoroacetate salt, 27.4mg, yield: 9.7%) properties: a white solid.
HPLC：90.63％@214nm，96.38％@254nmLCMS：MS m/z(ESI):407.1[M+H]
1 H NMR(400MHz,DMSO-d 6 )δ11.11(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＝5.2Hz,2H),3.20(s,3H),1.79(s,3H),1.76(s,3H).
Example 6
(2- ((5-chloro-1- (2-methoxyethyl) -1H-pyrazol-4-yl) pyrimidin-4-yl) Pyrazol-4-yl) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000641
Compound D20 (100mg, 0.57mmol) and intermediate D12 (150mg, 0.475mmol) were dissolved in isopropanol (20 mL) at room temperature, trifluoroacetic acid (1.1g, 9.5mmol) was added to the reaction system, and the reaction mixture was heated to 120 ℃ and stirred for reaction for 3 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000642
) Purification gave the desired product E6 (hydrochloride, 29.4mg, yield: 13.6%), property: a white solid.
HPLC：98.83％@214nm，99.17％@254nm
LCMS：MS m/z(ESI):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-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) Yl) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000643
Compound D21 (180mg, 0.32mmol) 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 completion of the reaction was checked by LCMS, the reaction was concentrated under reduced pressure and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000644
Figure GDA0003667177710000651
) Purification gave pure product E7 (HCl salt, 16mg, yield: 32%), property: a white solid;
HPLC：99.084％@214nm，99.750％@254nm
LCMS：MS m/z(ESI):460.4[M+H]
1 H NMR(400MHz,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＝12.8Hz,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-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) Amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000652
Compound D23 (100mg, 0.5 mmol) and intermediate D12 (131mg, 0.42mmol) were dissolved in isopropanol (20 mL) at room temperature, trifluoroacetic acid (960mg, 8.4 mmol) was added to the reaction system, and the reaction mixture was heated to 120 ℃ and stirred for reaction for 3 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000653
) Purification gave the desired product E8 (hydrochloride, 35.9mg, yield: 17.8%) trait: a white solid.
HPLC：98.84％@214nm，99.86％@254nm
LCMS：MS m/z(ESI):481.3[M+H]
1 H NMR(400MHz,MeOH-d 4 )δ8.6-8.1(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 Yl) phenyl) dimethylphosphine oxide (E9)
Figure GDA0003667177710000661
Compound D25 (100mg, 0.39mmol) and intermediate D12 (148mg, 0.47mmol) were dissolved in isopropanol (10 mL) at room temperature, and after trifluoroacetic acid (889.2mg, 7.8mmol) was added to the reaction system, the reaction was stirred at 120 ℃ 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 purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000662
) Purification gave the desired product E9 (trifluoroacetate salt, 22.4mg, yield: 11%), property: a white solid.
HPLC：99.20％@214nm，99.82％@254nm
LCMS：MS m/z(ESI):421.2[M+H]
1 H NMR(400MHz,MeOD)δ8.47(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) Phenyl) dimethyl phosphine oxide
Figure GDA0003667177710000663
Compound D27 (155mg, 1.0mmol) and intermediate D12 (158mg, 0.5mmol) were dissolved in isopropanol (20 mL) at room temperature, trifluoroacetic acid (1.14g, 10.0mmol) was added to the reaction system, and the reaction solution was heated to 120 ℃ and stirred for reaction for 3 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000664
Figure GDA0003667177710000671
) Purification gave the title product E10 (trifluoroacetate salt, 35.1mg, yield: 16.2%), trait: a white solid.
HPLC：96.49％@214nm，98.48％@254nm
LCMS：MS m/z(ESI):435.4[M+H]
19 F NMR (376.5 MHz, methanol-d 4): delta-77.21(s)
1 H NMR(400MHz,MeOH-d 4 )δ8.6-8.0(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
Figure GDA0003667177710000672
Compound D28 (30mg, 0.056 mmol) was dissolved in tetrahydrofuran/methanol (1 mL/1 mL) at room temperature, and sodium borohydride (11mg, 0.28mmol) was added to the reaction solution under ice-bath conditions. The reaction solution was stirred at room temperature for two hours. After the reaction was completed by LCMS, the reaction mixture was poured into saturated aqueous ammonium chloride (5 mL), the mixture was extracted with ethyl acetate (10 mLX), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000673
) Purification gave pure product E11 (trifluoroacetate, 20mg, yield: 50%). The characteristics are as follows: a colorless oil.
HPLC：93.04％(214nm)，93.66％(254nm)
LCMS:Rt:0.70min；MS m/z(ESI):504.5[M-H]。
1 H NMR(400MHz,MeOD)δ8.43(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- Yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000681
Compound D12 (95mg, 0.30mmol), compound D29 (50mg, 0.27mmol) and NH were added at room temperature 4 Cl (72mg, 1.35mmol) was dissolved in EtOH (3 mL) and the reaction was stirred at 100 ℃ for 10 h. After completion of the reaction by LCMS, the reaction was cooled to room temperature and the mixture was filtered to remove NH 4 Cl, the filtrate was concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000682
) Purification gave the desired product E12 (HCl salt, 15mg, yield: 12%). The characteristics are as follows: a white solid;
HPLC：99.574％@214nm，99.705％@254nm
LCMS：MS m/z(ESI):461.4[M+H]
1 H NMR(400MHz,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-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000683
Compound D31 (80mg, 0.386mmol) and intermediate D12 (101mg, 0.322mmol) were dissolved in isopropanol (20 mL) at room temperature, trifluoroacetic acid (734mg, 6.44mmol) was added to the reaction system, and the reaction was heated to 120 ℃ and stirred for 3 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000684
Figure GDA0003667177710000691
) Purification gave the desired product E13 (hydrochloride, 3.1mg, yield: 2%), property: a white solid.
HPLC：97.42％@214nm，97.22％@254nm
LCMS：MS m/z(ESI):487.4[M+H]
1 H NMR(400MHz,MeOH-d 4 )δ8.56-7.97(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) pyrimidine-4-) Yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000692
Compound D32 (36mg, 0.12mmol) and intermediate D29 (43mg, 0.24mmol) were dissolved in isopropanol (10 mL) at room temperature, and trifluoroacetic acid (274mg, 2.4mmol) was added to the reaction system; after the addition, the reaction solution was heated to 120 ℃ and stirred to react for 3 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000693
) Purification gave the title product E14 (hydrochloride, 24.7mg, yield: 46.3%) behavior: a white solid.
HPLC：97.09％@214nm，96.73％@254nm
LCMS：MS m/z(ESI):477.4[M+H]
19 F NMR (376.5 MHz, methanol-d) 4 )δ161.97(s)
1 H NMR(400MHz,MeOH-d 4 )δ8.70-7.91(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＝11.6Hz,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) pyrimidine-4-) Yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000701
Compound D33 (100mg, 0.28mmol) and compound D29 (51mg, 0.28mmol) were dissolved in EtOH (10 mL) in a 100mL single-neck flask at room temperature, trifluoroacetic acid (5 mL) was added, and the reaction was heated to 80 ℃ and stirred overnight. LCMS to check reaction completion, reaction was concentrated under reduced pressure and residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000702
) Purification gave the title compound E15 (trifluoroacetate salt, 8mg, yield: 10%,), properties: a white solid.
HPLC：99.487％@214nm,99.739％@254nm
LCMS：MS m/z(ESI):507[M+H]
19 F NMR(376.5MHz,DMSO)δ74.35(s)
1 H NMR(400MHz,DMSO)δ11.11(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) pyrimidine-4-) Yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000703
Compound D34 (80mg, 0.25mmol), compound D29 (45mg, 0.25mmol) and ammonium chloride (110mg, 2.5 mmol) were dissolved in ethanol (5 mL) at room temperature. The reaction mixture was stirred at 100 ℃ for 12 hours. After LCMS check that the reaction had been completed the reaction mixture was concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000704
Figure GDA0003667177710000711
) Purification gave pure product E16 (hydrochloride, 6mg, yield: 5%), property: yellow solid
HPLC：97.65％(214nm)，99.39％(254nm)
LCMS:Rt:1.04min；MS m/z(ESI):457.4[M+H]。
1 H NMR(400MHz,MeOD)δ8.66(brs,1H),7.72–7.64(m,1H),7.57(s,2H),7.36(t,J＝7.2Hz,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-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) Pyrimidin-4-yl) amino) phenyl) phosphine oxides
Figure GDA0003667177710000712
Compound D35 (50mg, 0.1699 mmol), compound D29 (30mg, 0.1699 mmol) and NH were added at room temperature 4 Cl (27mg, 0.507mmol) was dissolved in EtOH (3 mL) and the reaction was heated to 100 ℃ and stirred for 8 hours. LCMS detects that the reaction is complete, the mixture is filtered, and the filtrate is decompressed and concentrated; the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000713
) Purification to obtain pure target product E17 (HCl salt, 15mg, yield: 20%). The characteristics are as follows: an oily liquid;
HPLC：98.434％@214nm，98.466％@254nm
LCMS：MS m/z(ESI):441.4[M+H]
1 H NMR(400MHz,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＝11.2Hz,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-pyrazol-1-yl) piperidin-1-yl acetamide
Figure GDA0003667177710000721
Compound D37 (104mg, 0.2mmol), ammonium chloride (22mg, 0.4 mmol), HATU (114mg, 0.3mmol) was dissolved in DMF (2 mL) at room temperature. The reaction mixture was stirred at room temperature for 12 hours. After the reaction had been completed by LCMS, the reaction mixture was poured into water (5 mL). The mixture was extracted with ethyl acetate (10mL X4), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000722
) Purification gave pure product E18 (hydrochloride, 20mg, yield: 20%), property: pale yellow solid
HPLC：99.79％(214nm)，99.89％(254nm)
LCMS:Rt:0.66min；MS m/z(ESI):517.5[M-H]。
1 H NMR(400MHz,MeOD)δ8.57(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).
Examples 19 and 20
Dimethyl (2- ((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) -5- (trifluoromethyl) pyrimidine Pyridin-4-yl) amino) phenyl) phosphine oxide and dimethyl (2- ((4- ((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) Radical) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) phosphine oxide
Figure GDA0003667177710000723
Compound D40 (450mg, 0.76mmol) was dissolved in 4M HCl/dioxane (20 mL) at room temperature, the reaction was stirred at room temperature for 2h, and LCMS detected reaction completion. The reaction solution was concentrated under reduced pressure to obtain a target crude product (400 mg), and 50mg of the crude product was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000724
Figure GDA0003667177710000731
) Purification gave pure target E19 (hydrochloride, 12.3mg, yield: 24.6%) behavior: a white solid.
HPLC：99.58％@214nm，99.85％@254nm,Rt:6.543min
LCMS：MS m/z(ESI):494.4[M+H],Rt:1.094min
1 H NMR (400 MHz, methanol-d) 4 )δ8.44(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 )δ64.68(s).
Compound D41 (420mg, 0.76mmol) was dissolved in 4M HCl/dioxane (20 mL) at room temperature, the reaction was stirred at room temperature for 2h, and the reaction was determined to be complete by LCMS. The reaction solution was concentrated under reduced pressure to obtain a target crude product (400 mg), and 50mg of the crude product was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000732
) Purification gave pure target E20 (hydrochloride, 14.6mg, yield: 29.2%,) trait: a white solid.
HPLC：99.79％@214nm，99.93％@254nm,Rt:5.625min
LCMS：MS m/z(ESI):494.5[M+H],Rt:0.915min
19 F NMR (376.5 MHz, methanol-d) 4 )δ63.652(s).
1 H NMR(400MHz,MeOH-d 4 )δ8.44(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
Figure GDA0003667177710000733
Compound E7 (180mg, 0.36mmol), 1-fluoro-2-iodoethane (75mg, 0.43mmol), DIEA (140mg, 1.08mmol) was dissolved in DMF (3 mL) at room temperature. The reaction mixture was stirred at room temperature for 12 hours. LCMS check reaction was complete and reaction was directly purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000741
) Purification gave pure product E21 (30 mg, yield: 17%). The characteristics are as follows: white solid
HPLC：97.37％(214nm)，97.47％(254nm)
LCMS:Rt:0.76min；MS m/z(ESI):506.4[M+H]。
1 H NMR(400MHz,DMSO)δ11.18(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(400MHz,DMSO-d 6 )δ11.11(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＝5.2Hz,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) pyrimidine-4-) Yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000742
Compound D43 (150mg, 0.73mmol) and intermediate D12 (158mg, 0.5mmol) were dissolved in isopropanol (30 mL) at room temperature, and trifluoroacetic acid (1.14g, 10.0 mmol) was added to the reaction system; after the addition, the reaction solution was heated to 120 ℃ and stirred for reaction for 3 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000743
Figure GDA0003667177710000751
) Purification gave the desired product E22 (hydrochloride salt, 14.6mg, yield: 6.2%) properties: yellow solid.
HPLC：95.92％@214nm，97.59％@254nm
LCMS：MS m/z(ESI):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＝12.0Hz,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 (376.5 MHz, 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 Pyridin-4-yl) amino) phenyl) dimethylphosphineoxide (cis and trans)
Figure GDA0003667177710000752
A solution of compound D49 (200mg, 0.42mmol) in methanol (5 mL) was cooled in an ice bath, and then sodium borohydride (160mg, 0.42mmol) was slowly added to the reaction solution; after the addition was complete, the reaction was carried out at room temperature for 2 hours. After completion of the reaction, the reaction was quenched by adding saturated aqueous ammonium chloride (10 mL), the mixture was concentrated under reduced pressure, and the residue was extracted with methylene chloride/methanol (20/1, 10 mL. Times.3). The organic phases are combined and concentrated directly under reduced pressure, and the residue is purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000753
) Purifying to obtain pure target product E23 (40mg, 20% yield). The characteristics are as follows: a white solid.
HPLC：99.51％@214nm，99.26％@254nm
LCMS：MS m/z(ESI):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 other isomer was of poor purity and purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000754
Figure GDA0003667177710000761
) Purification gave pure target E24 (trifluoroacetate salt, 10mg, yield: 5%) property: white solid.
HPLC：96.45％@214nm，96.83％@254nm
LCMS：MS m/z(ESI):475.4[M+H]
1 H NMR(400MHz,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- (trifluoro) Methyl) pyrimidin-4-yl) amino) yl dimethyl phosphine oxide and (2- ((4- ((1- (1- (2-hydroxyethyl) piperidin-4-yl) -5-) methyl-1H-pyrazol-4-yl) amino) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000762
Compound E19 (hydrochloride, 100mg, 0.2mmol) and bromoethanol (38mg, 0.3mmol) were dissolved in MeCN (30 mL) at room temperature, and K was added to the reaction system 2 CO 3 (110mg, 0.8 mmol) and then the reaction mixture was heated to 80 ℃ and stirred for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with acetonitrile (10mL X2), the filtrate was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000763
) Purification gave the desired product E25 (16.9 mg, yield: 15.5%) properties: a white solid.
HPLC：95.79％@214nm，95.88％@254nm,Rt:6.415min.
LCMS：MS m/z(ESI):538.5[M+H],Rt:1.096min
1 H NMR (400 MHz, methanol-d) 4 )δ8.23(s,1H),8.14-8.10(m,1H),7.51-7.46(m,2H),7.29(t,J＝8.0Hz,1H),7.11-7.08(m,1H),4.14(br,1H),3.71(t,J＝6.0Hz,2H),3.14-3.11(m,2H),2.59(t,J＝6.0Hz,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 (376.5 MHz, methanol-d) 4 )δ-63.388(s).
Compound E20 (100mg, 0.2mmol) and bromoethanol (38mg, 0.3mmol) were dissolved in acetonitrile (30 mL) at room temperature, and K was added to the reaction system 2 CO 3 (110mg, 0.8 mmol) and then the reaction mixture was heated to 80 ℃ and stirred for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with acetonitrile (10mL X2), the filtrate was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000771
) Purification yielded the objective product E26 (23.4 mg, yield: 21.4%) properties: a white solid.
HPLC：98.56％@214nm，98.34％@254nm，Rt:5.485min.
LCMS：MS m/z(ESI):538.6[M+H],Rt:0.916min.
19 F NMR (376.5 MHz, methanol-d) 4 )δ-61.933(s).
1 H NMR(400MHz,MeOH-d 4 )δ8.23(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＝6.0Hz,2H),3.14-3.11(m,2H),2.59(t,J＝6.0Hz,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]Pyrimidine-4- Yl) aminophenyl) dimethylphosphine oxide
Figure GDA0003667177710000772
Compound D59 (50mg, 0.25mmol) and intermediate D12 (79.0 mg, 0.25mmol) were dissolved in EtOH (5 mL) at room temperature, and NH was added to the reaction system 4 After Cl (81mg, 1.5mmol), the reaction mixture was heated to 80 ℃ and stirred for reaction for 16 hours. After the reaction was completed, 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 purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000773
Figure GDA0003667177710000781
) Purification gave the desired product E27 (trifluoroacetate salt, 15.3mg, yield: 16%) properties: a white solid.
HPLC：95.66％@214nm，97.91％@254nm
LCMS：MS m/z(ESI):477.4[M+H]
1 H NMR(400MHz,MeOH-d 4 )δ7.96-8.45(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＝12.0Hz,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) Pyridin-4-yl) amino) phenyl) dimethylphosphineoxide (cis and trans)
Figure GDA0003667177710000782
Dimethylamine hydrochloride (257mg, 3.2mmol) and triethylamine (646mg, 6.4mmol) were added to dry dichloromethane (20 mL) at room temperature and stirred for half an hour at room temperature, then Compound D49 (150mg, 0.32mmol) was added, followed by stirring for half an hour at room temperature, then sodium cyanoborohydride (340mg, 1.6mmol) was added and the reaction was stirred at room temperature overnight. After completion of the reaction, the reaction solution was washed with a saturated aqueous ammonium chloride solution (10mL X2), and the organic phase was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000783
purification gave the desired product E28 (trifluoroacetate salt, 40mg, yield: 25% as a white solid) and product E29 (trifluoroacetate salt, 40mg, yield: 25% as a white solid).
E28:
HPLC：99.24％@214nm，99.00％@254nm,Rt:3.983min
LCMS：MS m/z(ESI):501[M+H],Rt:1.104min
19F NMR (376.5 MHz, methanol-d 4) delta-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/z(ESI):501[M+H],Rt:1.124min
19F NMR(376.5MHz,DMSO-d6)δ-74.70(s)
1 H NMR(400MHz,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)]Pyrimidine-4- Radical) aminophenyl) dimethyl phosphine sulfide
Figure GDA0003667177710000791
Compound E12 (100mg, 0.15mmol) and Lawson's reagent (122mg, 0.3mmol) were dissolved in toluene (5 mL) at room temperature, and the reaction mixture was heated to 110 ℃ and stirred for 4 hours. After the reaction was completed, the reaction solution was cooled to room temperature, a yellow solid precipitated, the suspension was filtered, the cake was washed with toluene (5 mL), the filtrate was concentrated under reduced pressure, and the residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000792
) Purification gave the desired product E30 (trifluoroacetate salt, 20.7mg, yield: 29.1%). The characteristics are as follows: a white solid.
HPLC：97.38％@214nm，97.75％@254nm
LCMS：MS m/z(ESI):477.3[M+H]
1 H NMR(400MHz,DMSO-d 6 )δ8.20–7.66(m,2H),7.46(brs,2H),4.40(brs,1H),4.06(dd,J＝11.4,4.1Hz,2H),3.58(t,J＝11.6Hz,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 Pyridin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000793
The compound D51 (50mg, 0.15mmol), the compound D29 (28mg, 0.15mmol) and NH were reacted at room temperature 4 Cl (24mg, 0.45mmol) was dissolved in EtOH (5 mL), and the reaction was heated to 100 ℃ and stirred for 2 hours. LCMS detection reaction complete, mixThe mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000801
) Purification is carried out to obtain a pure target product E31 (hydrochloride, 15mg, yield: 12%), property: a white solid;
HPLC：98.731％@214nm，98.291％@254nm
LCMS：MS m/z(ESI):467.4[M+H]
1 H NMR(400MHz,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-1- (2-azazole [ 3.5)]Non-7-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000802
Trifluoroacetic acid (300mg, 2.42mmol) was added to a solution of compound D56 (300mg, 0.48mmol) in dichloromethane (10 mL) at room temperature, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and one third of the reaction solution was purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000803
) Purification is carried out to obtain a pure target product E32 (trifluoroacetate, 40mg, yield: 25% white solid).
HPLC：95.34％@233nm，95.85％@254nm
LCMS：MS m/z(ESI):520.3[M+H]
1 H NMR(400MHz,MeOD)δ：8.30(br，1H),8.10(br,1H),7.69-7.64(m,2H),7.53(br,1H),7.35(t,J＝7.2Hz,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-pyranyl) Azol-4-yl) amino) pyrimidine-5-carboxylic acid ethyl ester
Figure GDA0003667177710000811
Compound D57 (710mg, 2.0mmol), intermediate D29 (400mg, 2.2mmol), ammonium chloride (1.1g, 20mmol) and ethanol (20 mL) were each added at room temperature to a 100mL three-necked flask. The mixture was heated to reflux and the reaction stirred for 16 hours. After the reaction was completed, the reaction liquid was cooled to room temperature, then the reaction liquid was concentrated under reduced pressure, and the residue was purified by column chromatography (eluent gradient DCM/methanol =80: 1-40) to obtain the objective product E33 (600 mg, yield: 60%, white solid).
HPLC：96.12％@214nm，97.10％@254nm
LCMS：MS m/z(ESI):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＝7.2Hz,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
Figure GDA0003667177710000812
1-methyl-4- (5-methyl-4-amino-1H-pyrazol-1-yl) piperidine (100mg, 0.32mmol), (2- ((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (120mg, 0.64mmol) and trifluoroacetic acid (2 mL) were dissolved in ethanol (3 mL) at room temperature. The reaction solution was heated to 100 ℃ and stirred for 12 hours. LCMS to check the reaction was complete, the reaction was cooled and concentrated under reduced pressure and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000813
) And purifying to obtain a pure target product E34 (3mg, yield:1.9%, white solid).
HPLC：87.09％@214nm，86.80％@254nm
LCMS：MS m/z(ESI):474.4[M+H]
1 H NMR(400MHz,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) pyrimidine-4- Yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000821
(S) -5-methyl-4-amino-1- (tetrahydrofuran-3-yl) -1H-pyrazole (50.0mg, 0.3mmol), (2- ((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (104.0mg, 0.33mmol) was dissolved in isopropanol (2 mL) at room temperature, trifluoroacetic acid (680.0 mg, 6.0mmol) was then added, the mixture was heated to 120 ℃ and stirred for 4 minutes, and the reaction was checked to be complete by LCMS. The reaction solution was cooled to room temperature and concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000822
) Purification gave 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(400MHz,DMSO)δ11.19(s,1H),8.51(brs,1H),8.06(s,1H),7.54(dd,J＝12.0Hz,8.0Hz,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-) 1H-pyrazole-5-carbonitrile
Figure GDA0003667177710000823
5-cyano-4-amino-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazole (150mg, 0.78mmol) and (2- ((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (250mg, 0.78mmol) were dissolved in ethanol (10 mL) at room temperature, and then ammonium chloride (530mg, 10mmol) was added to the solution; the reaction system is heated to 80 ℃ under the protection of argon, and stirred to react overnight. After completion of the reaction, the reaction solution was cooled to room temperature, the mixture was filtered, the filtrate was concentrated under reduced pressure, and the obtained residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000831
) Purification to obtain the final product E36 (trifluoroacetate). The product was dissolved in dichloromethane/methanol (20/1, 5 mL), the solution was washed with saturated aqueous sodium bicarbonate (2mL X3) and water (2mL X2), and the residue was dried to give the desired product E36 (40mg, yield 11%, white solid) in free form
HPLC：97.21％@214nm，96.07％@254nm,Rt:8.807min.
LCMS：MS m/z(ESI):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 Radical) phenyl) bis (methylphosphorus oxide
Figure GDA0003667177710000832
(2- ((2-chloro-5-isopropylpyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (30mg, 0.09mmol), and 5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-amine (17mg, 0.09mmol) and ammonium chloride (24mg, 0.45mmol) were dissolved in ethanol (5 mL) at room temperature, and the reaction mixture was heated to 100 ℃ and stirred for reaction for 10 hours. LCMS check reaction complete. The reaction solution was cooled to room temperature and then filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000833
Figure GDA0003667177710000841
) Purification gave pure product E37 (hydrochloride, 8.1mg, yield: 12%, yellow solid);
HPLC：98.624％@214nm，98.489％@254nm
LCMS：MS m/z(ESI):469.5[M+H]
1 H NMR(400MHz,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 compound
Figure GDA0003667177710000842
(2- ((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (230mg, 0.7 mmol), 1-tert-butyl-1H-pyrazol-4-amine (100mg, 0.7 mmol) and ammonium chloride were dissolved in ethanol (3 mL) at room temperature. The reaction solution was heated to 85 ℃ and stirred to react for 12 hours. LCMS to check the reaction was complete, the reaction solution was cooled and filtered, and the filtrate was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000843
) And purifying to obtain a pure target product E38 (170mg, yield:58% light red solid
HPLC：99.31％@214nm，99.56％@254nm。
LCMS:Rt:1.327min；MS m/z(ESI):419.4[M+H]。
1 H NMR(400MHz,DMSO)δ11.11-10.82(m,1H),9.21(s,1H),8.15(s,1H),7.75(br,1H),7.61(dd,J＝16.0Hz,8.0Hz,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
Figure GDA0003667177710000851
1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-amine (80mg, 0.48mmol) was dissolved in ethanol (10 mL) at room temperature, followed by addition of (2- ((5-bromo-2-chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (173mg, 0.48mol) and ammonium chloride (260mg, 4.8mmol). The reaction solution was heated to 80 ℃ and stirred overnight, LCMS detected complete reaction, reaction solution was cooled and filtered, filtrate was concentrated under reduced pressure, and residue was purified by high performance liquid chromatography (depoling gradient:
Figure GDA0003667177710000852
) Purification afforded product E39 (25 mg, yield: 20%, off-white solid).
HPLC：98.296％@214nm，98.785％@254nm
LCMS：MS m/z(ESI):491.3[M+H]
1H NMR(400MHz,DMSO)δ11.20-10.20(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- Pyrazol-1-yl) piperidin-1-yl) -2-hydroxy-1-ones
Figure GDA0003667177710000853
(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, 0.6 mmol) were dissolved in N, N-dimethylformamide (2 mL) at room temperature. The reaction solution was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was diluted with water (5 mL), followed by extraction with dichloromethane/methanol (10, 10mL X3), the organic phases were combined and dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000854
Figure GDA0003667177710000861
) And purifying to obtain a pure product E40 (15mg, 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)δ11.18(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 Pyridin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000862
(2- ((5-chloro-2- ((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (100mg, 0.2 mmol), 1,1-difluoro-2-iodoethane (58mg, 0.3 mmol), and diisopropylethylamine (72mg, 0.6 mmol) were dissolved in N, N-dimethylformamide (2 mL) at room temperature. The reaction solution was heated to 70 ℃ and stirred to react for 12 hours. LCMS to check the reaction was complete, the reaction mixture was cooled and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000863
) Purification was carried out to obtain a pure product E41 (formate, 15mg, yield:15% white solid).
HPLC：94.86％@214nm，96.38％@254nm
LCMS:MS m/z(ESI):524.3[M+H]
1 H NMR(400MHz,MeOD)δ8.42(brs,1H),8.17(brs,1H),7.96(s,1H),7.55(dd,J＝16.0Hz,8.0Hz,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-pyrazol-1-yl) piperidin-1-yl) acetic acid formate salt
Figure GDA0003667177710000871
Methyl 2- (4- (4- ((5-chloro-4- ((2- (dimethylphosphoryl) phenyl) amino) pyrimidin-2-yl) amino) -5-methyl-1H-pyrazol-1-yl) piperidin-1-yl) acetate (50mg, 0.096 mmol) and lithium hydroxide (7mg, 0.29mmol) were dissolved in methanol (2 mL) and water (1 mL) at room temperature. The reaction solution was stirred at room temperature for 12 hours. LCMS check reaction was complete, the reaction was filtered and the filtrate was purified directly by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000872
) And purifying to obtain a pure target product E42 (formate, 40mg, yield:80% colorless oily liquid)
HPLC：97.90％@214nm，98.25％@254nm。
LCMS:Rt:0.83min；MS m/z(ESI):518.4[M+H]。
1 H NMR(400MHz,MeOD)δ8.58(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-pyranyl) Azol-4-yl) amino) pyrimidine-5-carbonitriles
Figure GDA0003667177710000873
(2- ((5-bromo-2- ((5-methyl-1- (tetrahydro-2H-pyran-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (100mg, 0.19mmol), zinc cyanide (22mg, 0.19mmol), tris (dibenzylideneacetone) dipalladium (17.3mg, 0.02mmol) and bis (diphenylphosphino) ferrocene (10.5mg, 0.02mmol) were mixed and added to N, N-dimethylformamide (5 mL) at room temperature, and the reaction system was heated to 100 ℃ under a nitrogen atmosphere and stirred for 8 hours. After completion of 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 and dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000881
) Purification gave the product E43 (20mg, yield:23.5% white solid
HPLC：99.068％@214nm，99.421％@254nm
LCMS：MS m/z(ESI):452.4[M+H]
1 H NMR(400MHz,MeOD-d 4 ):δ8.50-8.07(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-1- (2-methyl-2-azo [3.5]]Nonyl-7-yl) -1H-pyrazol-4-yl) amino group Pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000882
(2- ((5-chloro-1- (2-azaoxazolo [3.5] nonan-7-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (MED-SIM-2-157) (200mg, 0.38mmol) and a solution of aqueous formaldehyde (100mg, 37% in water, 1.2 mmol) in methanol (20 mL) were stirred at room temperature for 16 hours, then sodium triacetoxyborohydride (270mg, 1.2mmol) was added, and the reaction was allowed to continue stirring at room temperature for 30 minutes. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, the residue was dispersed in water (10 mL) and methylene chloride (10 mL), the two phases were separated, and the aqueous phase was extracted with methylene chloride (20mL X3). The organic phases are combined, dried over anhydrous sodium sulfate and filtered, the filtrate is concentrated under reduced pressure, and the residue is purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000883
Figure GDA0003667177710000891
) Purification gave a pure target E44 (the TFA salt obtained was free with potassium carbonate) (50mg, yield:23% white solid
HPLC：97.66％@233nm，98.87％@254nm
LCMS：MS m/z(ESI):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) Dimethyl phosphine oxide
Figure GDA0003667177710000892
1- (2-methoxyethyl) -1H-pyrazol-4-amine (47.0mg, 0.33mmol) and (2- ((5-modifying-2-chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (100.0mg, 0.28mmol) were dissolved in isopropanol (5 mL) at room temperature, and trifluoroacetic acid (319.2mg, 2.80mmol) was further added to the reaction system; the reaction solution was heated to 120 ℃ and stirred to react for 3 hours. After the reaction was completed, the reaction solution was cooled and filtered, the filter cake was washed with isopropyl alcohol (20 ml), the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000893
) Purification to obtain the target product E45 (29.9mg, yield:23% brown solid).
HPLC：99.62％@214nm，99.78％@254nm
LCMS：MS m/z(ESI):467.2[M+H]
1 H NMR(400MHz,DMSO-d 6 )：δ9.26(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
Figure GDA0003667177710000901
(2- ((5-bromo-2- ((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide hydrochloride (50mg, 0.095 mmol) and N, N-diisopropylethylamine (49mg, 0.38mmol) were dissolved in N, N-dimethylformamide (15 mL) at room temperature, and 1-fluoro-2-iodoethane (25mg, 0.143mmol) was added dropwise to the reaction system; the reaction solution was heated to 70 ℃ and stirred to react for 16 hours. LCMS to check the reaction was complete, the reaction was cooled to room temperature and diluted with water (50 mL), the mixture was extracted with ethyl acetate (20mL X4), the organic phases were combined and washed with saturated sodium chloride solution (50mL X2), then dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000902
) Purification gave the target product E46 (4.3 mg, yield: 7.9%, white solid).
HPLC：95.2％@214nm，95.8％@254nm
LCMS：MS m/z(ESI):536.5[M+H]
1 H NMR(400MHz,MeOH-d 4 )δ8.14(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- Pyrazol-1-yl) piperidin-1-yl) propionitrile
Figure GDA0003667177710000903
(2- ((5-chloro-2- ((5-methyl-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (72mg, 0.11mmol) was dissolved in N, N, -dimethylformamide (10 mL) at room temperature, and acrylonitrile (10mg, 0.11ummol) and sodium carbonate (34mg, 0.32mmol) were added; the reaction solution was stirred at 65 ℃ overnight, LCMS detected complete reaction, the reaction solution was cooled and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (depoling gradient:
Figure GDA0003667177710000911
) Purification afforded product E47 (25 mg, yield: 20%, off-white solid).
HPLC：93.920％@214nm，94.278％@254nm
LCMS：MS m/z(ESI):513.4[M+H]
1 H NMR(400MHz,MeOD)δ8.42(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) dimethylphosphine oxide
Figure GDA0003667177710000912
(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 (5 mL) at room temperature, methylvinylsulfone (13mg, 0.124mol) and sodium carbonate (26mg, 0.248mmol) were further added, and the reaction mixture was heated to 65 ℃ and stirred overnight. LCMS to check the reaction was complete, the reaction solution was cooled and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000913
) Purification afforded product E48 (25 mg, yield: 20%, off-white solid).
HPLC：93.512％@214nm，96.920％@254nm
LCMS：MS m/z(ESI):566.4[M+H]
1 H NMR(400MHz,MeOD)δ8.43(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-modified-2- ((1- (1- (2,2-difluoroethyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000921
(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 (15 mL) at room temperature, and 1,1-difluoro-2-iodoethane (30mg, 0.15mmol) was added dropwise to the reaction system; the reaction solution was heated to 70 ℃ and stirred to react for 16 hours. LCMS to check the reaction was complete, the reaction was cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (20mL X4), the organic phases combined, washed with saturated sodium chloride solution (50mL X2), dried and filtered over anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000922
) Purification gave the target product E49 (4.6 mg, yield: 6.8%, lot # white solid).
HPLC：93.9％@214nm，95.5％@254nm
LCMS：MS m/z(ESI):554.3[M+H]
1 H NMR(400MHz,MeOH-d 4 )δ8.14-7.80(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) pyrimidine-4- Radical) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000923
(2- ((5-chloro-2- ((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (120mg, 0.27mmol) was dissolved in N, N, -dimethylformamide (10 mL), followed by addition of (methylsulfonyl) ethylene (30mg, 0.27mol) and sodium carbonate (60mg, 0.54mmol). The reaction was heated to 60 ℃ and stirred overnight. LCMS to check the reaction was complete, the reaction was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000931
) Purification gave the desired product E50 (25 mg, yield: 20%). The characteristics are as follows: a white-like solid.
1 H NMR(400MHz,DMSO)δ11.5-10.5(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/z(ESI):552.4[M+H]
Example 51
(2- ((5-chloro-2- ((1- (1- (2-methoxyethyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4-) Radical) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000932
(2- ((5-chloro-2- ((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (100mg, 0.22mmol), 1-bromo-2-methoxyethane (31mg, 0.22mmol), and N, N-diisopropylethylamine (28mg, 0.22mmol) were dissolved in N, N-dimethylformamide (3 mL). The reaction was stirred at room temperature for 10 hours and LCMS detected complete reaction. The reaction solution was directly subjected to preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710000933
Figure GDA0003667177710000943
) Purification gave pure product E51 (25.5 mg, yield: 23%), character: a white solid.
1 H NMR(400MHz,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/z(ESI):504.4[M+H]
Example 52
(2- ((5-chloro-1- (1- (2- (methylsulfonyl) ethyl) piperidin-4-yl) -1H-pyrazol-4-yl) Amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000941
(2- ((5-chloro-1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (90mg, 0.19mmol), methanesulfonylethylene (24mg, 0.23mmol) and diisopropylethylamine (49mg, 0.38mmol) were dissolved in N' N-dimethylformamide (2 mL) at room temperature. The reaction mixture was stirred at 60 ℃ for 2 hours. LCMS to check reaction completion, the reaction mixture was concentrated under reduced pressure and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000942
) Purification was carried out to obtain a pure product E52 (45 mg, yield: 35%), properties: beige solid
1 H NMR(400MHz,MeOD)δ8.37(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＝8.0Hz,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:0.96min；MS m/z(ESI):586.4[M+H]。
Example 53
2- ((5-chloro-2- ((1- (2- (2-methoxyethyl) -2-azaspiro [ 3.5)]Nonan-7-yl) -1H-pyrazol-4-yl Amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000951
2- (2-methoxyethyl) -7- (4-amino-1H-pyrazol-1-yl) -2-azaspiro [3.5] nonane (100mg, 0.37mmol), (2- ((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (120mg, 0.37mmol) and ammonium chloride (196mg, 3.7 mmol) were dissolved in ethanol (5 mL) at room temperature, the reaction system was heated to reflux, and the reaction was stirred for 16 hours. After completion of the reaction, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000952
) Purification gave the pure desired product E53 (30.9 mg, yield: 15.4%), property: white solid
1 H NMR(400MHz,DMSO-d6)：δ11.30-10.75(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/z(ESI):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) pyrimidinePyridine- 4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000953
(2- ((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (3) (80mg, 0.25mmol), 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 (5 mL) at room temperature, and the reaction solution was heated to 80 ℃ and stirred for reaction overnight. After completion of the reaction, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000954
Figure GDA0003667177710000961
) Purification gave the pure product E54 (11 mg, yield: 8.8%), property: white solid
1 H NMR(400MHz,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/z(ESI):500.2[M+H]
Example 55
2- ((5-chloro-2- ((1- (2- (2-fluoroethyl) -2-azaspiro [ 3.5)]Nonan-7-yl) -1H-pyrazol-4-yl) amino group Pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000962
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 (450mg, 1.43mmol) and trifluoroacetic acid (1600mg, 14.3mmol) were dissolved in isopropanol (10 mL) at room temperature, the reaction system was heated to reflux, and the reaction was stirred for 16 hours. After completion of the reaction, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000963
) Purification gave the pure target product E55 (trifluoroacetate salt, 62.2mg, yield: 8.1%), property: white solid
1 H NMR(400MHz,MeOD)：δ8.09(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/z(ESI):532.4[M+H]
Example 56
(2- ((5-chloro-2- ((1-methylcyclopentano [ c)]Pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Radical) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710000971
2-methyl-5- (4-amino-1H-pyrazol-1-yl) cyclopentylalkyl [ c ] pyrrolidine (430mg, 2.07mmol), (2- ((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (650mg, 2.07mmol) was dissolved in ethanol (10 mL) at room temperature, and ammonium chloride (1.3g, 20.7mmol) was added; the reaction solution was heated to 80 ℃ and stirred overnight. LCMS to check reaction completion, the reaction was concentrated under reduced pressure and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000972
) Purification gave the desired product E56 (103.7 mg, yield: 10.6%). The characteristics are as follows: a white-like solid.
1 H NMR(400MHz,MeOD)δ8.40-8.05(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/z(ESI):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 Pyridin-4-yl) amino) phenyl) dimethylphosphine sulfide
Figure GDA0003667177710000973
(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-dimethyloxirane (447mg, 6.2mmol) and potassium carbonate (427mg, 3.1mmol) were dissolved in ethanol/water (6 mL/1.5 mL) at room temperature. The reaction solution was heated to 110 ℃ in a microwave reactor and reacted for 20 minutes with stirring. LCMS to check the reaction was complete, the reaction mixture was concentrated under reduced pressure and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000981
) Purification gave a pure product E57 (30 mg, yield: 21%), properties: white solid
1 H NMR(400MHz,DMSO)δ9.39-9.27(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:1.16min；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
Figure GDA0003667177710000982
2-methyl-6- (4-amino-1H-pyrazol-1-yl) -2-azaspiro [3.3] heptane (150mg, 0.78mmol), (2- ((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (8) (24msg, 0.78mmol) was dissolved in ethanol (10 mL) at room temperature, ammonium chloride (560g, 7.8mmol) was added thereto, and the reaction mixture was heated to 80 ℃ and stirred overnight. LCMS to check reaction completion, reaction filtered, filtrate concentrated under reduced pressure, residue purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000983
) Purification gave the target product E58 (19.4 mg, yield: 20%), property: a white-like solid.
1 H NMR(400MHz,DMSO)δ11.35-11.10(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/z(ESI):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-) Yl) amino) phenyl) dimethylphosphine oxide (example 59, single isomer 1: rt:5.078min; example 60 Single isomerization Body 2: rt:5.106 min)
Figure GDA0003667177710000991
1-methyl-4- (3- (4-amino-1H-pyrazol-1-yl) cyclobutyl) piperazine (106mg, 0.45mmol), (2- ((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (157mg, 0.5mmol) and ammonium chloride (240mg, 4.5mmol) were dissolved in isopropanol (3 mL) at room temperature. The reaction mixture was stirred at 80 ℃ for 12 hours. LCMS to check reaction completion, the reaction mixture was concentrated under reduced pressure and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710000992
) Purification to obtain pure product E59 (single isomer 1: rt:5.078 min) (31.5 mg, yield: 12%), properties: a white solid; and E60 (single isomer 2 rt: white solid.
E59 (single isomer 1
1 H NMR(400MHz,MeOD):δ8.25(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:0.855min；MS m/z(ESI):515.4[M+H]。
E60 (single isomer 2
1 H NMR(400MHz,MeOD):δ8.37(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:0.889min；MS m/z(ESI):515.4[M+H]。
Example 61
2- ((5-chloro-2- ((1- (2-morphinylethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) benzene Yl) dimethyl phosphine oxide
Figure GDA0003667177710001001
(2- ((5-chloro-2- ((1- (2-chloroethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl phosphine oxide (70mg, 0.16mmol), morpholine (27mg, 0.32mmol) and diisopropylethylamine (66mg, 0.48mmol) were dissolved in N, N-dimethylformamide (5 mL), and the reaction solution was stirred at room temperature for 10 hours and was detected to be complete by LCMS. The reaction solution was filtered, and the filtrate was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001002
) Purification gave pure product E61 (trifluoroacetate, 43.4mg, yield: 57.1%), property: a white solid.
1 H NMR(400MHz,MeOD)：δ8.12(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/z(ESI):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
Figure GDA0003667177710001003
(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 (5 mL) at room temperature; the reaction solution was heated to 70 ℃ and stirred to react for 24 hours. LCMS check reaction complete and reaction mixture concentrated. The residue was then purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001004
Figure GDA0003667177710001011
) Purification gave E62 (trifluoroacetate salt, 10mg, yield: 5%) as a solid product: a white solid.
1 H NMR(400MHz,MeOD):δ8.24(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:1.30min；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 Pyridin-4-yl) amino) phenyl) dimethylphosphineSulfide compound
Figure GDA0003667177710001012
(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-dimethyloxirane (120mg, 1.67mmol), and potassium carbonate (115mg, 0.85mmol) were dissolved in ethanol/water (2 mL/0.5 mL) at room temperature; the reaction solution was heated to 110 ℃ in a microwave reactor and reacted for 20 minutes with stirring. LCMS check reaction complete and reaction mixture concentrated. The residue was then purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001013
) Purification gave E63 (trifluoroacetate, 10mg, yield: 6%) as a product with the following properties: a white solid.
1 H NMR(400MHz,MeOD):δ8.24(brs,1H),7.99-7.40(m,6H),4.18(brs,1H),3.90(d,J＝11.2Hz,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:1.20min；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) Phenyl) dimethyl phosphine oxide
Figure GDA0003667177710001021
(2- ((5-chloro-2- ((1- (2-chloroethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (70mg, 0.16mmol), pyrrolidine (6) (35mg, 0.49mmol) and diisopropylethylamine (63mg, 0.49mmol) were dissolved in N, N-dimethylformamide (5 mL). The reaction was stirred at room temperature for 10 hours and LCMS showed completion. The reaction solution was filtered, and the filtrate was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001022
) Purification gave pure product E64 (19.3 mg, yield: 26.4%), property: a white solid.
1 H NMR(400MHz,MeOD)：δ8.51-8.05(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/z(ESI):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
Figure GDA0003667177710001023
(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-dimethylformamide (4 mL) at room temperature, and the reaction was stirred at 80 ℃ for 16 hours; after completion of the reaction, the reaction solution was cooled to room temperature, the mixture was filtered, and the filtrate was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001024
Figure GDA0003667177710001031
) Purification was carried out to obtain a pure target product E65 (trifluoroacetate salt, 9.3mg, yield: 10%), property: a 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/z(ESI):510.5[M+H]
Example 66
(2- ((5-chloro-2- ((1- (2- (2,2-difluoroethyl) -2-azaspiro [ 3.5)]Nonan-7-yl) -1H-pyrazol-4-yl Amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710001032
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) were dissolved in isopropanol (5 mL) at room temperature, the reaction was heated to reflux, and stirred for reaction for 16 hours. After completion of the reaction, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001033
) Purification gave the pure target product E66 (trifluoroacetate salt, 6.0mg, yield: 11%), property: white solid
1 H NMR(400MHz,MeOD)：δ8.25-8.19(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/z(ESI):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
Figure GDA0003667177710001041
Triethylamine (12.1mg, 0.12mmol) was added to a solution of (2- ((2- ((1- (2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) direct) -5-chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine sulfide (60mg, 0.12mmol) and an aqueous formaldehyde solution (97mg, 37% aqueous solution, 1.2 mmol) in methanol (5 mL) at room temperature, and after a reaction time of 5 hours with stirring, sodium triacetoxyborohydride (101.8mg, 0.48mmol) was added, and the reaction mixture was further stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001042
) Purification gave the title compound E67 (trifluoroacetate salt, 20.1mg, yield: 33%), property: a white solid.
1 H NMR(400MHz,MeOD)：δ8.15(brs,1H),7.97-7.38(m,6H),4.20(d,J＝8.0Hz,1H),4.04(d,J＝8.0Hz,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/z(ESI):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) dimethylphosphine oxide
Figure GDA0003667177710001043
(2- ((2- ((1- (2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) amino) -5-chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (48.5mg, 0.1mmol), 1,1,1-trifluoro-2-iodoethane (41.8mg, 0.2mol) and diisopropylethylamine (24.4mg, 0.2mmol) were dissolved in acetonitrile (5 mL) at room temperature, and the reaction solution was heated to 80 ℃ and stirred for reaction for 16 hours. After completion of the reaction, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001051
) Purification gave the pure target product E68 (trifluoroacetate salt, 32mg, yield: 56%), property: white solid.
1 H NMR(400MHz,MeOD)：δ8.13(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/z(ESI):568.5[M+H]
Example 69
(2- ((5-chloro-2- ((1- (2- (2-hydroxy-2-methylpropyl) cyclopenta [ c)]Pyrrolidin-5-yl) -1H-pyridine Oxazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl)
Dimethyl phosphine oxide
Figure GDA0003667177710001052
(2- ((5-chloro-2- ((1- (cyclopenta [ c ] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (160mg, 0.30mmol), 2,2-dimethyloxirane (216mg, 3.0 mmol) and potassium carbonate (207mg, 5.0 mmol) were dissolved in ethanol/water (2 mL/0.5 mL) at room temperature. The reaction solution was heated to 110 ℃ in a microwave reactor and reacted for 20 minutes with stirring. LCMS to check the reaction was complete, the reaction mixture was concentrated under reduced pressure and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001053
) Purification to obtain pure product E69 (hydrochloride, 30mg, yield: 21%), property: white solid
1 H NMR(400MHz,DMSO)δ9.27(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:0.96min；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) pyrimidine-4- Radical) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710001061
(2- ((5-chloro-2- ((1- (cyclopenta [ c ] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (160mg, 0.30mmol), iodoethane (56mg, 0.36mmol) and diisopropylethylamine (116mg, 0.90mmol) were dissolved in N' -N-dimethylformamide (1 mL) at room temperature. The reaction mixture was stirred at 70 ℃ for 12 hours. LCMS check reaction complete and reaction mixture was concentrated under reduced pressure. The residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001062
) Purification was carried out to obtain a pure product E70 (14 mg, yield: 10%), properties: a white solid.
1 H NMR(400MHz,DMSO)δ9.28(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:0.96min；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) amine Yl) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710001063
(2- ((5-chloro-2- ((1- (cyclopenta [ c ] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (100mg, 0.21mmol), 1-fluoro-2-iodoethane (36mg, 0.21mmol) and N, N-isopropylethylamine (54mg, 0.42mmol) were dissolved in N, N-dimethylformamide (3 mL). The reaction was stirred at 100 ℃ for 8 hours, LCMS checked for completion, and the reaction was purified by hplc (eluent gradient:
Figure GDA0003667177710001071
) Purification gave pure product E71 (25.5 mg, yield: 20.4%), property: a white solid.
1 H NMR(400MHz,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/z(ESI):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) dimethylphosphine oxide
Figure GDA0003667177710001072
(2- ((5-chloro-2- ((1- (cyclopenta [ c ] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (150mg, 0.31mmol), 1,1-difluoro-2-iodoethane (60mg, 0.31mmol) and N, N-isopropylethylamine (54mg, 0.42mmol) were dissolved in acetonitrile (3 mL). The reaction was stirred at 70 ℃ for 8 hours, LCMS checked for completion, and the reaction was purified by hplc (eluent gradient:
Figure GDA0003667177710001073
) Purification gave pure product E72 (13.2 mg, yield: 7.8%), property: white solid.
1 H NMR(400MHz,MeOD)：δ8.10(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/z(ESI):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) dimethylphosphine oxide
Figure GDA0003667177710001081
(2- ((5-chloro-2- ((1- (cyclopenta [ c ] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (100mg, 0.21mmol), 1,1,1-trifluoro-2-iodoethane (48mg, 0.21mmol), and N, N-isopropylethylamine (54mg, 0.42mmol) were dissolved in acetonitrile (3 mL). The reaction was stirred at 70 ℃ for 8 hours, LCMS checked for completion, and the reaction was purified by hplc (eluent gradient:
Figure GDA0003667177710001082
) Purification gave pure product E73 (20.2 mg, yield: 20.4%), property: a white solid.
1 H NMR(400MHz,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/z(ESI):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
Figure GDA0003667177710001091
To a microwave tube were added in sequence (2- ((2- ((1- (2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) direct) -5-chloropyrimidin-4-yl) amino) phenyl) dimethyl phosphine sulfide (60mg, 0.12mmol), 2,2-dimethyloxirane (86.4mg, 1.2mol), potassium carbonate (66.2mg, 0.48mmol), and ethanol/water (4 mL/1 mL) under argon protection. The reaction solution was heated to 110 ℃ by microwave and stirred for 1 hour. After completion of the reaction, 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 high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001092
) Purification gave the pure target product E74 (10.6 mg, yield: 14.7% white solid
1 H NMR(400MHz,MeOD)：δ8.07(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) dimethylphosphine oxide
Figure GDA0003667177710001093
To a microwave tube were added in sequence (2- ((2- ((1- (2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) amino) -5-chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (150mg, 0.31mmol), 2,2-dimethyloxirane (89.3mg, 1.24mol), potassium carbonate (171mg, 1.24mmol) and ethanol/water (8 mL/2 mL) under argon protection. The reaction solution was heated to 110 ℃ by microwave and reacted for 1 hour. After completion of the reaction, 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 high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001094
Figure GDA0003667177710001101
) Purification gave the pure target product E75 (trifluoroacetate salt, 108mg, yield: 62.8%), property: white solid
1 H NMR(400MHz,MeOD)：δ8.35-8.10(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/z(ESI):558.2[M+H]
Example 76
(2- ((5-chloro-2- ((1- (2- (2-methoxyethyl) cyclopenta [ c)]Pyrrolidin-5-yl) -1H-pyrazole-4- Yl) amino) pyrimidin-4-yl) Amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710001102
(2- ((5-chloro-2- ((1- (cyclopenta [ c ] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (160mg, 0.30mmol), bromoethyl ethane (50mg, 0.36mmol) and diisopropylethylamine (116mg, 0.90mmol) were dissolved in N, N-dimethylformamide (1 mL) at room temperature. The reaction solution was heated to 70 ℃ and stirred to react for 12 hours. LCMS to check reaction completion, the reaction mixture was concentrated under reduced pressure and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001103
) Purification was carried out to obtain pure product E76 (7 mg, yield: 5%), properties: white solid
1 H NMR(400MHz,DMSO)δ9.26(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:1.00min；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-pyrazol-4-yl Amino) pyrimidin-4-yl) amino) phenyl) dimethyl phosphine sulfide
Figure GDA0003667177710001111
(2- ((2- ((1- (2-azaspiro [3.5] non-7-yl) -1H-pyrazol-4-yl) direct) -5-chloropyrimidin-4-yl) amino) phenyl) dimethyl phosphine sulfide (60mg, 0.12mmol), 1-bromo-2-methoxyethane (33.1mg, 0.24mol) and diisopropylethylamine (29.3mg, 0.24mmol) were dissolved in acetonitrile (5 mL) at room temperature, and then the reaction solution was heated to 80 ℃ and stirred for reaction for 16 hours. After completion of the reaction, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001112
) Purification gave the pure target product E77 (trifluoroacetate salt, 8.7mg, yield: 13%), property: white solid
1 H NMR(400MHz,MeOD)：δ8.07(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/z(ESI):560.2[M+H]
Example 78
(2- ((5-chloro-1- (2- (2-methoxyethyl) -2-azaspiro [ 3.5)]Nonan-7-yl) -1H-pyri-dine Azol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710001113
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 (3 mL). The reaction was stirred at 100 ℃ for 8 hours, LCMS detected complete reaction, the reaction cooled to room temperature, filtered and the filtrate purified by preparative high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710001114
Figure GDA0003667177710001121
) Purification was carried out to obtain a pure target product E78 (trifluoroacetate salt, 9.4mg, yield: 7.1%), property: a white solid.
1 H NMR(400MHz,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/z(ESI):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
Figure GDA0003667177710001122
(2- ((5-chloro-2- ((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (100mg, 0.22mmol), 2,2,2-trifluoroethyl triflate (102mg, 0.44mmol) and diisopropylethylamine (142mg, 1.1 mmol) were dissolved in acetonitrile (4 mL) at room temperature, and the reaction was stirred at 70 ℃ for 12 hours; after completion of the reaction, the reaction solution was cooled and concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001123
) Purification was carried out to obtain a pure target product E79 (18.5 mg, yield: 16%), trait: a 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/z(ESI):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
Figure GDA0003667177710001131
(2- ((5-chloro-2- ((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (100mg, 0.22mmol), 1,1-difluoro-2-iodoethane (84mg, 0.44mmol), and potassium carbonate (121mg, 0.88mmol) were added to N, N-dimethylformamide (4 mL) at room temperature, and the reaction was stirred at 80 ℃ for 12 hours; after completion of the reaction, the reaction liquid was cooled and concentrated under reduced pressure, and the residue was purified by preparative chromatography using high performance liquid chromatography (eluent gradient:
Figure GDA0003667177710001132
) Purification was carried out to obtain a pure target product E80 (40 mg, yield: 36%), trait: a 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＝56.0Hz,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/z(ESI):510.4[M+H]
Example 81
2- ((5-chloro-2- ((1- (2- (dimethylamino) ethyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) Phenyl) dimethylphosphine oxide
Figure GDA0003667177710001133
N, N-dimethyl-2- (4-amino-1H-pyrazol-1-yl) ethylamine (100mg, 0.65mmol), (2- ((2,5-dichloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (205mg, 0.65mmol), trifluoroacetic acid (661mg, 6.5 mmol) was dissolved in ethanol (5 mL) at room temperature, and the reaction solution was stirred at 50 ℃ for 12 hours; after completion of the reaction, the reaction solution was cooled and concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001134
Figure GDA0003667177710001141
) Purification was carried out to obtain a pure target product E81 (trifluoroacetate salt, 93mg, yield: 33%), property: yellow solid.
1 H NMR(400MHz,DMSO-d6):δ11.19-10.79(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/z(ESI):434.4[M+H]
Example 82
(2- ((5-chloro-2- ((1- (1- (2-hydroxy-2-methylpropyl) piperidin-4-yl) -5-methyl-1H-pyrazol-4-yl) Amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine sulfide
Figure GDA0003667177710001142
(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-dimethyloxirane (125mg, 1.7 mmol), and potassium carbonate (120mg, 0.87mmol) were dissolved in ethanol/water (2 mL/0.5 mL) at room temperature. The reaction solution was heated to 110 ℃ in a microwave reactor and reacted for 20 minutes with stirring. LCMS check reaction complete and reaction mixture was concentrated under reduced pressure. The residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001143
) Purification was carried out to obtain pure product E82 (6 mg, yield: 6%), properties: a white solid.
1 H NMR(400MHz,DMSO)δ9.38(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:1.16min；MS m/z(ESI):548.4[M+H]。
Example 83
(2- ((5-chloro-2- ((1- (2-hydroxy-2-methylpropyl) piperidine)-4-yl) -1H-pyrazol-4-yl) amino) pyrimidine- 4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710001151
(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-dimethyloxirane (162mg, 2.2mmol), and potassium carbonate (155mg, 1.1mmol) were dissolved in a mixture of ethanol (3 mL) and water (0.75 mL) at room temperature, and the reaction was heated to 110 ℃ with a microwave and stirred for 20 minutes. After completion of the reaction, the reaction solution was cooled and concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001152
) Purification was carried out to obtain a pure target product E83 (58 mg, yield: 50%), property: a 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/z(ESI):518.4[M+H]
Example 84
(2- ((5-chloro-2- ((1- (1- (2- (dimethylamino) ethyl) piperidin-4-yl) -1H-pyrazol-4-yl) amino) Pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710001153
(2- ((5-chloro-2- ((1- (piperidin-4-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (100mg, 0.22mmol), 2-chloro-N, N-dimethylethylamine hydrochloride (32mg, 0.22mmol), sodium carbonate (70mg, 0.66mmol) and iodine sodium (7 mg, 0.044mmol) were added to N' N-dimethylformamide (4 mL) at room temperature, and the reaction was stirred at 80 ℃ for 12 hours; after completion of the reaction, the reaction solution was cooled and concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001161
) Purification was carried out to obtain a pure target product E84 (58 mg, yield: 50%), property: a 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/z(ESI):517.6[M+H]
Example 85
2- ((5-chloro-2- ((1- (2-methyl-cyclopenta [ c ]]Pyrrole-5-acyl) -1 h-pyrazole-4-acyl) amino) pyrimidines Pyridine-4-acyl) amino) phenyl) dimethylphosphinothio
Figure GDA0003667177710001162
At room temperature, (2- ((5-chloro-2- ((1- (cyclopenta [ c ] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl phosphine sulfide (80mg, 0.116mmol) was dissolved in methanol (4 mL), triethylamine was added dropwise to make the solution alkaline, a 30% aqueous formaldehyde solution (300mg, 3.2mmol) was added, and the reaction was stirred at room temperature for 12 hours; then sodium triacetoxyborohydride (339mg, 1.6mmol) is added and stirred for 1 hour at room temperature; after completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001163
) Purification is carried out to obtain a pure target product E85 (34 mg, yield: 42%), property: a 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/z(ESI):502.4[M+H]
Example 86
(2- ((5-bromo-2- ((1-methylcyclopentano [ c)]Pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Radical) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710001171
2-methyl-5- (4-amino-1H-pyrazol-1-yl) cyclopentylalkyl [ c ] pyrrolidine (62mg, 0.3 mmol), (2- ((5-bromo-2-chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (108mg, 0.3 mmol) and ammonium chloride (160mg, 3.0 mmol) were dissolved in isopropanol (3 mL) at room temperature. The reaction mixture was stirred at 80 ℃ for 12 hours. After completion of the reaction by LCMS, the reaction mixture was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001172
) Purification was carried out to obtain a pure product E86 (15 mg, yield: 10%), properties: a white solid.
1 H NMR(400MHz,DMSO)δ10.90-10.31(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:0.93min；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) dimethylphosphine oxide
Figure GDA0003667177710001173
(2- ((2-chloro-5- (trifluoromethyl) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (119mg, 0.34mmol), (2- ((5-chloro-2- ((1- (cyclopenta [ c ] pyrrolidin-5-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (70mg, 0.34mmol) and ammonium chloride (180mg, 3.4mmol) were dissolved in ethanol (5 mL) at room temperature, the reaction was heated to reflux, and the reaction was stirred for 16 hours. After completion of the reaction, the reaction solution was cooled to room temperature and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001181
) Purification gave the pure target product E87 (19.6 mg, yield: 11.1% white solid
1 H NMR(400MHz,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/z(ESI):520.1[M+H]
Example 88
(2- ((5-chloro-2- ((1- (3- (4- (2-methoxyethyl) piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) Yl) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710001182
(2- ((5-chloro-2- ((1- (3- (piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl phosphine oxide (40mg, 0.079mmol), bromoethyl methyl ether (21mg, 0.158mmol) and N, N-isopropylethylamine (30mg, 0.237mmol) were dissolved in N, N-dimethylformamide (3 mL). The reaction was stirred at room temperature for 10 hours, LCMS detected reaction was complete, and the reaction was directly purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001183
) Purification gave pure product E88 (8 mg, yield: 18.2%), character: a white solid.
1 H NMR(400MHz,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＝5.6Hz,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/z(ESI):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-) Yl) amino) phenyl) dimethylphosphine oxide (example 89, single unknown configurational isomer, rt:3.313min; in the case of the embodiment 90, the method comprises the following steps, Singleunknown configurational isomer, rt 3.949min)
Figure GDA0003667177710001191
Tert-butyl 3- (4- ((5-chloro-4- ((2- (dimethylphosphinoxy) phenyl) amino) pyrimidin-2-yl) amino) -1H-pyrazol-1-yl) piperidine-1-carboxylate (400mg, 0.73mmol) was dissolved in hydrogen chloride in dioxane (4 m,5 mL) and methanol (0.5 mL) at room temperature, and the reaction mixture was reacted at room temperature for 0.5 hour. Completion of the reaction was checked by LCMS, the reaction solution was concentrated under reduced pressure, then dissolved in N, N-dimethylformamide (5 mL), bromoethyl methyl ether (100mg, 0.73mmol) and N, N' -diisopropylethylamine (283mg, 2.19mmol) were added, and after the reaction mixture was reacted at room temperature for 10 hours, completion of the reaction was checked by LCMS, and the reaction solution was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001192
) Purification was carried out to obtain a pure racemic product (150 mg, yield: 40.8%) properties: yellow solid, chiral separation of target product:
the racemic product (150mg, 0.3mmol) was subjected to chiral separation (separation conditions:
instrument Signal Waters-SFC80
Chromatographic column type IA-H (2.5X 25cm,10 μm)
The mobile phase A is the supercritical carbon dioxide,
mobile phase B ethanol/ammonia
B =70/30, flow rate: 70g/min
Detection wavelength of 214nm
Column temperature 25 deg.C
Column pressure 100 bar)
The target product E89 (single unknown configurational isomer, rt:3.313 min) (54.93 mg, yield: 36.6%, property: yellow solid) and the target product E90 (single unknown configurational isomer, rt:3.949 min) (59.64 mg, yield: 39.766%, property: yellow solid) were obtained.
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/z(ESI):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/z(ESI):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 Pyridin-4-yl) amino) phenyl) dimethylphosphine oxide (example 91, unknown single isomer 1, rt:2.784min; in accordance with an embodiment 92 of the present invention, unknown single isomer 2, rt:4.559 min)
Figure GDA0003667177710001201
(2- ((5-chloro-2- ((1- (piperidin-3-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (310mg, 0.7 mmol), 2,2-dimethyloxirane (62mg, 0.84mmol), and potassium carbonate (193mg, 1.4 mmol) were dissolved in a mixed solvent of ethanol/water (4/1, 10 mL) at room temperature, and the system was heated to 110 ℃ by microwave for reaction for 3 hours. After LCMS detection of reaction completion, the reaction was filtered and the filtrate was purified directly by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001202
) Purification gave the racemic target product (160 mg, yield: 20%), property: white solid
LCMS:Rt:1.168min；MS m/z(ESI):518[M+H]。
The racemic target product (160mg, 0.3mmol) was separated by supercritical chiral hplc (separation conditions:
instrument Signal Waters-SFC80
Type of chromatographic column IA-H (2.5 × 25cm,10 μm)
The mobile phase A is the supercritical carbon dioxide,
mobile phase B ethanol/ammonia
B =60/40, flow rate: 70g/min
Detection wavelength of 214nm
Column temperature 25 deg.C
Column pressure 100 bar) to give two single isomers:
e91 (unknown single isomer 1,rt 2.784min) (10 mg, yield: 6%), properties: off-white solid
1 H NMR(400MHz,MeOD)δ8.20(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/z(ESI):518.4[M+H]
E92 (unknown single isomer 2, rt 4.559min) (50 mg, yield: 30%), properties: off-white solid
1 H NMR(400MHz,MeOD)δ8.20(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/z(ESI):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
Figure GDA0003667177710001212
(2- ((2- ((1- (3-azaspiro [5.5] undecan-9-yl) -1H-pyrazol-4-yl) amino) -5-chloropyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (30mg, 0.05mmol), 2,2-dimethyloxirane (36mg, 0.5 mmol), and potassium carbonate (42mg, 0.25mmol) were dissolved in ethanol/water (2 mL/0.5 mL) at room temperature. The reaction solution was heated to 110 ℃ in a microwave reactor and stirred for 20 minutes. LCMS to check the reaction was complete, the reaction mixture was concentrated under reduced pressure and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001211
) Purification to obtain pure product E93 (hydrochloride, 6mg, yield: 20%), property: a white solid;
1 H NMR(400MHz,MeOD)δ8.25(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:0.98min；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
Figure GDA0003667177710001221
(2- ((5-chloro-2- ((1- (3- (piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl phosphine sulfide (33mg, 0.064 mmol), 2,2-dimethyloxirane (46mg, 0.64mmol), and potassium carbonate (45mg, 0.32mmol) were dissolved in a mixed solution of ethanol (2 mL) and water (0.5 mL) at room temperature, and the reaction was heated to 110 ℃ with microwave and stirred for 20 minutes; after completion of the reaction, the reaction solution was cooled to room temperature, then concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001222
) Purification is carried out to obtain a pure target product E94 (2 mg, yield: 5%), property: a white solid.
1 H NMR(400MHz,MeOD–d4)δ：8.08(s,1H),7.85(brs,2H),7.65(t,J＝8.0Hz,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/z(ESI):589.1[M+H]
Example 95
2- ((5-chloro-2- ((1- (3- (4-ethylpiperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidine-4- Yl) amino) phenyl) dimethyl phosphine sulfide
Figure GDA0003667177710001223
(2- ((5-chloro-2- ((1- (3- (piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl phosphine sulfide (33mg, 0.064 mmol) was dissolved in ethanol (3 mL) at room temperature, triethylamine was added dropwise to make the solution basic, 1M acetaldehyde tetrahydrofuran solution (1.3ml, 1.28mmol) was added, and the reaction was stirred at room temperature for 12 hours; then adding sodium triacetoxyborohydride (136mg, 0.64mmol), reacting, and continuing stirring for 1 hour at room temperature; after completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001231
) Purification was carried out to obtain a pure target product E95 (trifluoroacetate salt, 4mg, yield: 11%), property: a white solid.
1 H NMR(400MHz,MeOD-d4)：δ8.14(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/z(ESI):545.4[M+H]
Example 96
(2- ((5-chloro-2- ((1- (3- (4- (2-methoxyethyl) piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) Yl) pyrimidin-4-yl) amino) phenyl) dimethylphosphine sulfide
Figure GDA0003667177710001232
(2- ((5-chloro-2- ((1- (3- (piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethyl phosphine sulfide (33mg, 0.064mmol), bromoethyl methyl ether (11mg, 0.077mmol), diisopropylethylamine (41mg, 0.32mmol) was dissolved in N' N-dimethylformamide (3 mL) at room temperature, and the reaction was stirred at 50 ℃ for 12 hours; after completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001233
) Purification is carried out to obtain a pure target product E96 (2 mg, yield: 5%), property: white solid.
1 H NMR(400MHz,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＝5.6Hz,2H),3.34(s,3H),3.03(br,1H),2.62(t,J＝5.6Hz,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/z(ESI):575.5[M+H]
Example 97
2- ((5-chloro-2- ((1- (1-methylpiperidin-3-yl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) benzene Yl) dimethyl phosphine oxide
Figure GDA0003667177710001241
(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 (4 mL) at room temperature, triethylamine was added dropwise to make the solution basic, a 30% aqueous formaldehyde solution (1.3g, 14.4 mmol) was added, and the reaction was stirred at room temperature for 12 hours; sodium triacetoxyborohydride (916mg, 4.32mmol) was added thereto, and stirring was continued at room temperature for 1 hour. After completion of the reaction, the reaction was concentrated under reduced pressure, and the residue was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001242
) Purification is carried out to obtain a pure target product E97 (110 mg, yield: 34%), property: a white solid.
1 H NMR(400MHz,DMSO-d6)：δ11.30-10.65(m,1H),9.26(s,1H),8.14(m,2H),7.62(br,1H),7.54(t,J＝8.0Hz,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/z(ESI):460.3[M+H]
Example 98
(2- ((5-chloro-2- ((1- (3- (4-ethylpiperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidine-4-) Yl) amino) phenyl) dimethylphosphine oxide
Figure GDA0003667177710001243
In a 100mL single vial was added (2- ((5-chloro-2- ((1- (3- (piperazin-1-yl) cyclobutyl) -1H-pyrazol-4-yl) amino) pyrimidin-4-yl) amino) phenyl) dimethylphosphine oxide (80mg, 0.16mmol) and dissolved in ethanol (5 mL), followed by addition of a solution of acetaldehyde in tetrahydrofuran (0.64mL, 0.64mmol), after stirring the compound for 10 hours, sodium borohydride triacetate (67mg, 0.32mmol) was added, the reaction mixture was allowed to react for 1 hour at room temperature, and the reaction was complete by LCMS. The reaction was diluted with acetonitrile (2 mL) and purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001251
) Purification was carried out to obtain pure product E98 (4 mg, yield: 4.7%), properties: a 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/z(ESI):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) dimethylphosphine oxide
Figure GDA0003667177710001252
(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 (5 mL), 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, the reaction was detected to be complete by LCMS, and the reaction liquid was purified by high performance liquid preparative chromatography (eluent gradient:
Figure GDA0003667177710001253
) Purification gave pure product E99 (22 mg, yield: 20.4%), property: a 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/z(ESI):528.4[M+H]
LCMS：MS m/z(ESI):463.1[M+H]。
Test example 1: enzymatic EGFR phosphorylation inhibition assay
Experimental materials:
HTRF KinEASE-TKkit was purchased from CisBio (France).
The 384-well assay plate was purchased from greiner bio-one (Germany).
384-well source plate was purchased from LABCYTE (USA).
MgCl 2 ，MnCl 2 DTT, tritonX-100, HEPES, BSA were purchased from Sigma (USA).
EGFR WT-WT, EGFR WT-del19/T790M/C797S, EGFR WT-L858R/T790M/C797S, EGFR WT-del19/T790M, and EGFR WT-L858R/T790M are offered by Jiangsu Prodonce pharmaceutical Co., ltd.
An experimental instrument:
automatic micropore pipettor: precision PRC384U (BioTek, USA)
Nano-liter acoustic wave shift system:
Figure GDA0003667177710001261
HANDLE RS(LABCYTE，USA)
multi-label detection analyzer: envision Multilabel Reader (PerkinElmer, USA)
The experimental method comprises the following steps:
1. kinase buffer solution (2 mM HEPES pH7.5,0.01% TritonX-100,0.01% BSA,1mM DTT,1mM MnCl) was prepared 2 ，5mM MgCl 2 )。
2. Preparation of the compound. The final concentration of the compound tested was 10. Mu.M, formulated at 100-fold concentration, i.e., 1mM. Test compounds were diluted in 4-fold gradients using an automated micropore pipette (Precision PRC 384U) to obtain 10 dose concentrations from 1mM to 3.8nM, and test compounds were transferred to 384-well plates using Echo, 100nL of compound per well.
3. And (3) kinase reaction. The enzyme reaction solution was prepared using kinase buffer, 5. Mu.l of the enzyme reaction solution was transferred to 384-well reaction wells, and 5. Mu.l of kinase buffer was added to negative control wells. The mixture was shaken at 450rpm and incubated at room temperature for 10 minutes. The substrate solution was prepared using kinase buffer, 5. Mu.l of the substrate solution was transferred to reaction wells of 384-well plates to initiate the reaction, mixed by shaking at 450rpm, and incubated at room temperature for 60 minutes.
4. The reaction was terminated. Mu.l of a reaction stop solution (streptavidin-XL 665, TK Antibody-Cryptate) was added to a 384-well reaction plate, centrifuged at 1000rpm for 1 minute, left at room temperature for 60 minutes, and the Envision plate was read
5. And (6) fitting a curve. Data from the Envision program were reproduced for luminescence readings and the ratio of 665nM/615nM was converted to percent inhibition by the formula: percent inhibition = (max-signal ratio)/(max-min) × 100."min" is the ratio of fluorescence readings of a control sample reacted without enzyme; "max" is the ratio of fluorescence readings for samples with DMSO added as a control. Data were imported into MS Excel and curve-fitted using XLFit Excel add-in version 5.4.0.8: y = Bottom + (toptom)/(1 + (IC) 50 /X)^HillSlope)。
BaF of the Compound of the present invention 3 Inhibition of enzymatic Activity of (WT-WT) 50 Enzymatic Activity inhibition IC of BaF3 (WT-del 19/T790M/C797S) 50 ，BaF 3 Enzymatic activity inhibition IC of (WT-L858R/T790M/C797S) 50 ，BaF 3 Enzymatic Activity inhibition IC of (WT-del 19/T790M) 50 ，BaF 3 Enzymatic Activity inhibition IC of (WT-L858R/T790M) 50 As follows.
As can be seen from the following results, most of the compounds of the present invention have excellent inhibitory effects on four mutated enzymes, and the antiproliferative activity is below 100 nM. Most of the compounds of the invention have good selectivity on wild type EGFR, such as compounds E1', E26 and E30 for inhibiting the enzymatic activity of BaF3 (WT-del 19/T790M/C797S) 50 Is an enzymatic activity inhibitory IC for BaF3 (WT-WT) 50 Over 150 times of that of Brigatinib, and is obviously superior to Brigatinib.
Figure GDA0003667177710001262
Figure GDA0003667177710001271
Figure GDA0003667177710001281
Figure GDA0003667177710001291
Test example 2 cell anti-proliferation assay
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).
Cell
Figure GDA0003667177710001292
The 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 the Mediterranean crown Biotechnology (Taicang) Inc.
BaF3 medium: RPMI1640+10% of FBS +8ng/ml IL-3.
BaF3/EGFR-WT, baF3/EGFR-Del19/T790M/C797S and BaF3/EGFR-L858R/T790M/C797S media: RPMI1640+10% FBS.
Plate reading instrument: enVision (Perkinelmer, USA).
The experimental method comprises the following steps:
cell concentration was adjusted to 2000 BaF per well in 96-well plates 3 ，BaF 3 /EGFR-WT，BaF 3 EGFR-Del19/T790M/C797S or BaF 3 Per EGFR-L858R/T790M/C797S cells, 90. Mu.l volume, 1000 Xtest compound stock solution was prepared in DMSO, and test compound was diluted 3-fold in gradient (total 9 concentrations) in DMSO and 100-fold in the corresponding cell culture medium, 10. Mu.l (10 Xconcentration) of test compound (three duplicate wells per compound concentration) was added per well, 5. Mu.l CO 2 And cultured at 37 ℃ for 72 hours. After 72 hours, 50. Mu.l Cell Titer-Glo per well was added 2m on a plate shakerMix well in and incubate for 10min before reading with EnVision.
And (3) data analysis:
cell viability was plotted using GraphPad Prism version 5, and the IC was calculated by fitting a nonlinear regression model with a sigmoidal dose response 50 And (4) data.
Proliferation inhibition IC of most compounds of the invention in cell BaF3 (EGFR-del 19/T790M/C797S) 50 And BaF 3 Cell proliferation inhibition IC of (EGFR-L858R/T790M/C797S) 50 All have good performance and are obviously superior to the standard compound Brigatinib. The test results are shown in the following table:
Figure GDA0003667177710001301
Figure GDA0003667177710001311
the embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, or improvement made without departing from the spirit and principle of the present invention shall fall within the protection scope of the present invention.

Claims (8)

1. The following compounds or pharmaceutically acceptable salts thereof:
Figure FDA0003926886120000011
Figure FDA0003926886120000021
Figure FDA0003926886120000031
Figure FDA0003926886120000041
Figure FDA0003926886120000051
2. the compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the salt is a hydrochloride, trifluoroacetate or formate salt.
3. A pharmaceutical composition comprising a therapeutically effective amount of at least one compound of claim 1 or a pharmaceutically acceptable salt thereof.
4. The pharmaceutical composition of claim 3, further comprising a pharmaceutically acceptable excipient.
5. Use of at least one compound of claim 1 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of cancer.
6. The use of claim 5, wherein 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.
7. Use of at least one compound of claim 1 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a disease caused by EGFR mutation.
8. The use according to claim 7, wherein the EGFR mutation is one, two or more mutations selected from: (1) Del19; (2) T790M; (3) C797S; (4) L858R; (5) T790M; and (6) C797S.
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