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  • C07D487/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains four or more hetero rings
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  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
  • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
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  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
  • A61K31/529—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim forming part of bridged ring systems
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  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/66—Phosphorus compounds
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  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• • A—HUMAN NECESSITIES
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  • A61P35/00—Antineoplastic agents
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  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
  • C07D471/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed systems contains four or more hetero rings
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  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
  • C07D487/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains three hetero rings
  • C07D487/16—Peri-condensed systems
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  • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
  • C07D487/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains three hetero rings
  • C07D487/18—Bridged systems
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  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D498/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
  • C07D498/16—Peri-condensed systems
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  • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D498/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
  • C07D498/18—Bridged systems
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  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D498/22—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains four or more hetero rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
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  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
  • C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
  • C07F9/02—Phosphorus compounds
  • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
  • C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
  • C07F9/65616—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings containing the ring system having three or more than three double bonds between ring members or between ring members and non-ring members, e.g. purine or analogs

Definitions

• the present invention claims the priority of the Chinese patent application CN202111583828.0 with the application date of December 22, 2021, and the priority of the Chinese patent application CN202210098381.6 with the application date of January 27, 2022, and the application date of May 23, 2022.
• the priority of the Chinese patent application CN202210562007.7 dated December 07, 2022 is the priority of the Chinese patent application CN202211568373.X. This application cites part or all of the above-mentioned Chinese patent applications.
• the invention belongs to the field of medicine, and in particular relates to a pyrimidine heterocyclic compound, its preparation method and its application in medicine.
• CDK7 Cyclin-dependent kinase/cyclin complexes were identified as conserved components of the RNA polymerase II transcription machinery. Twenty mammalian CDKs currently exist. Among mammalian CDKs, CDK7 has robust kinase activity, and only CDK7 has dual functions of regulating cell cycle progression and transcription. In the cytosol, CDK7 exists as a heterotrimeric complex and is thought to function as a CDK1/2/4/6-activating kinase (CAK), whereby CDK7 contributes to the conservation of CDK1/2/4/6 Phosphorylation of residues is required for full catalytic CDK activity and cell cycle progression.
• CAK CDK1/2/4/6-activating kinase
• CDK7 forms the kinase core of the RNA polymerase II transcription factor complex and is responsible for phosphorylating the C-terminal domain (CTD) of RNA polymerase II, a necessary step for the initiation of gene transcription.
• CTD C-terminal domain
• CDK7 may serve as a therapeutic target for the treatment of many diseases and syndromes.
• CDK7 can interact with multiple transcription factors, cofactors, chromatin regulators and non-coding RNAs in the transcriptional regulatory region to regulate transcription. Mutations in these transcription factors, cofactors, chromatin regulators, or noncoding RNAs can lead to diseases such as cancer, autoimmune diseases, neurological disorders, developmental syndromes, diabetes, cardiovascular disease, and obesity.
• Some of these transcription factors control RNA polymerase II-mediated transcription initiation and elongation, and when their expression or function is altered, they can produce aggressive tumor cells (e.g. caused by c-Myc) or some forms of autoimmunity Sexuality (e.g. caused by AIRE).
• CDK7 kinase can promote the abnormal expression of some tumor-related transcription factors by regulating the overall transcription process, and promote tumor development by regulating the phosphorylation of key cell cycle kinases. More importantly, CDK7 more significantly regulates the expression of oncogenic transcription factors than other housekeeping genes in cancer cells. Inhibition of CDK7 can differentially affect the transcription of certain oncogenes and housekeeping genes, thus ensuring a therapeutic window. Transcription regulation and cell cycle regulation can be performed by regulating CDK7-mediated phosphorylation modification, which can be used to treat abnormal proliferation diseases including cancer. As a global regulator of transcription, CDK7 may also serve as a therapeutic target for the treatment of diseases such as inflammation, viral replication such as HIV, EBV, cancer and cardiac hypertrophy.
• diseases such as inflammation, viral replication such as HIV, EBV, cancer and cardiac hypertrophy.
• CDK7-selective inhibitors The high sequence and structural similarity of the kinase domains of CDK family members hampers the discovery of CDK7-selective inhibitors. Therefore, the development of selective CDK7 inhibitors is of great value for clinical application.
• the first aspect of the present invention provides a compound represented by formula (A) or a pharmaceutically acceptable salt thereof;
• Ring A is none; or Ring A is selected from the group consisting of 3 to 14 membered carbocycles, 3 to 14 membered heterocyclic rings, 5 to 12 membered heteroaryls, C 5-12 aryls; Ring A is unsubstituted or replaced by 1, 2, 3, 4 or 5 are substituted by groups selected from S1 group; the 3- to 14-membered heterocycle has 1, 2, 3 or 4 heteroatoms independently selected from nitrogen, oxygen and sulfur; The 5 to 12 membered heteroaryl has 1, 2, 3 or 4 heteroatoms independently selected from nitrogen, oxygen and sulfur;
• L 3 is none, C 1-6 alkylene, C 2-6 alkenylene or ring B (the structure is ); wherein, 1, 2, 3, 4 or 5 hydrogen atoms on the C 1-6 alkylene group can optionally be independently substituted by R 13 ; on the C 2-6 alkenylene group 1, 2, 3, 4 or 5 hydrogen atoms may optionally each be independently substituted by R 13 ;
• Ring B is selected from the group consisting of 3 to 14 membered carbocycle, 3 to 14 membered heterocyclic ring, 5 to 12 membered heteroaryl, C 5-12 aryl; Ring B is unsubstituted or replaced by 1, 2, 3, 4 or 5 are substituted by groups selected from S1 group; the 3 to 14 membered heterocyclic ring has 1, 2, 3 or 4 heteroatoms independently selected from nitrogen, oxygen and sulfur; the 5 to 12 membered Heteroaryl has 1, 2, 3 or 4 heteroatoms independently selected from nitrogen, oxygen and sulfur;
• the 5-12 membered heteroaryl has 1, 2, 3 or 4 heteroatoms independently selected from nitrogen, oxygen and sulfur; the C 5-12 aryl, 5-12 membered heteroaryl are each independently Unsubstituted or substituted by 1, 2, 3, 4 or 5 groups selected from S1 group;
• L2 is selected from the group consisting of:
• Each R 10 is independently hydrogen, C 1-6 alkyl, deuterated C 1-6 alkyl, -3 to 7-membered carbocyclyl, -C(O)-C 1-6 alkyl, -C(O )-C 1-4 alkyl-C 1-6 alkoxy, -C (O)-3 to 7-membered carbocyclyl, -C 1-4 alkyl-hydroxyl, -C 1-4 alkyl-cyanide Base, -C 1-4 alkyl-C 1-6 alkoxy, -C 1-4 alkyl -NHC (O)-C 1-6 alkyl, -C 1-4 alkyl - NHC (O) -C 1-4 alkyl-C 1-6 alkoxy, -C 1-4 alkyl-NHC(O)-3 to 7-membered carbocyclyl or -C 1-4 alkyl-NRR';
• Each 3 to 7 membered heterocyclic group independently has 1, 2 or 3 heteroatoms independently selected from nitrogen, oxygen and sulfur; each of the 3 to 7 membered carbocycles is independently unsubstituted or replaced by 1, Substituted by 2, 3, 4 or 5 groups selected from group S1;
• Each 3- to 7-membered carbocyclic group is independently unsubstituted or substituted by 1, 2, 3, 4 or 5 groups selected from group S1;
• each R 11 is independently hydrogen, C 1-6 alkyl or deuterated C 1-6 alkyl;
• Each R 12 is independently hydrogen, C 1-6 alkyl or deuterated C 1-6 alkyl;
• Each R 13 is independently hydrogen, cyano, hydroxyl, halogen, C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, -3 to 7-membered carbocyclyl, -C 0-6 alkylene-NRR', -C 1-6 alkylene-hydroxyl or -C 0-6 alkylene-cyano;
• Each R 14 is independently hydrogen, cyano, hydroxyl, halogen, C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, -3 to 7-membered carbocyclyl, -C 0-6 alkylene-NRR', -C 1-6 alkylene-hydroxyl or -C 0-6 alkylene-cyano;
• each m1 is each independently 1, 2, 3, 4, 5 or 6;
• Each m2 is independently 0 or 1;
• Each m3 is independently 0 or 1;
• each m4 is independently 1 or 2;
• each m5 is independently 0, 1, 2, 3, 4, 5 or 6;
• each m6 is independently 0, 1, 2, 3, 4, 5 or 6;
• Each m7 is independently 1 or 2;
• R 1 is hydrogen, halogen, cyano, C 1-6 alkyl or 3 to 14 membered carbocyclyl; said C 1-6 alkyl, 3 to 14 membered carbocyclyl are independently unsubstituted or replaced by 1 , 2, 3, 4 or 5 groups selected from the group substituted: oxo, hydroxyl, halogen, C 1-6 alkyl, halogenated C 1-6 alkyl;
• R 2 is hydrogen, halogen, cyano, C 1-6 alkyl or 3 to 14 membered carbocyclyl; said C 1-6 alkyl, 3 to 14 membered carbocyclyl are independently unsubstituted or replaced by 1 , 2, 3, 4 or 5 groups selected from the group substituted: oxo, hydroxyl, halogen, C 1-6 alkyl, halogenated C 1-6 alkyl;
• X 2 is N or C (R 3 );
• X 1 is N or C (R 4 );
• Y is N or C (R 4 );
• Z is N or C (R 3 );
• Each R is independently hydrogen, halogen, cyano, C 1-6 alkyl or halogenated C 1-6 alkyl;
• Each R is independently hydrogen, halogen, hydroxy, cyano, -C 2-4 alkenylene-phenyl, -C 2-4 alkynylene-phenyl, -S(O)-OH, -S( O) 2 -OH, -S-(C 1-6 alkyl), C 1-6 alkyl, -OC 1-6 alkyl, -C 1-6 alkylene -OC 1-6 alkyl, - OC 1-6 alkylene-OC 1-6 alkylene, -C 0-6 alkylene-NRR', -C 0-6 alkylene-C(O)OH, -C 0-6 alkylene -C(O)-C 1-6 alkyl, -C 0-6 alkylene-C(O)-NRR', -C 0-6 alkylene-NR-C(O)-C 1-6 Alkyl, -C 0-6 alkylene-S(O) 2 -C 1-6 alkyl, -C 0-6
• the groups of the S1 group include: oxo, halogen, hydroxyl, cyano, C 1-6 alkyl, -OC 1-6 alkyl, -C 1-6 alkylene-OC 1-6 alkyl, -OC 1-6alkylene -OC 1-6alkylene , -C 0-6alkylene -NRR', -C 0-6alkylene -C(O)OH, -C 0-6alkylene -C(O)-C 1-6 alkyl, -C 0-6 alkylene-C(O)-NRR', -C 0-6 alkylene-NR-C(O)-C 1- 6 alkyl, -C 0-6 alkylene-S(O) 2 -C 1-6 alkyl, -C 0-6 alkylene-S(O) 2 -NRR', -C 0-6 alkylene Alkyl-NR-S(O) 2 -C 1-6 alkyl, -C 0-6
• R and R' are each independently hydrogen, C 1-6 alkyl or deuterated C 1-6 alkyl; or R and R' optionally form a 3 to 14-membered heterocyclic group together with the nitrogen atom connected to them or 5 to 12-membered heteroaryl; wherein, the heterocyclic group and the heteroaryl each independently contain 0, 1 or 2 heteroatoms selected from N, O, and S in addition to the existing nitrogen atom;
• two hydrogen atoms on any same carbon atom on the C 0-6 alkylene group may also optionally be substituted by a 3-7 membered carbocyclic ring or a 3-7 membered heterocyclic spirocyclic ring at the same time ;
• two hydrogen atoms on any same carbon atom on the C1-6 alkylene group may optionally be simultaneously substituted by a 3- to 7-membered carbocyclic ring or a 3- to 7-membered heterocyclic spirocyclic ring .
• R is hydrogen, halogen, cyano, C 1-4 alkyl, or a 3- to 7-membered carbocyclyl; the C 1-4 alkyl, 3- to 7-membered carbocyclyl are independently Unsubstituted or substituted by 1, 2, 3, 4 or 5 groups selected from the group consisting of oxo, hydroxyl, halogen, C 1-3 alkyl, halo C 1-3 alkyl.
• R is hydrogen, halogen, cyano, C 1-4 alkyl (e.g. methyl), haloC 1-4 alkyl (e.g. trifluoromethyl), 3 to 7 membered carbocycle group or halogenated 3- to 7-membered carbocyclyl.
• R 2 is hydrogen, halogen, cyano, C 1-4 alkyl, or 3 to 7 membered carbocyclyl; said C 1-4 alkyl, 3 to 7 membered carbocyclyl are independently Unsubstituted or substituted by 1, 2, 3, 4 or 5 groups selected from the group consisting of oxo, hydroxyl, halogen, C 1-3 alkyl, halo C 1-3 alkyl.
• R is hydrogen, halogen, cyano, C 1-4 alkyl (e.g. methyl), haloC 1-4 alkyl (e.g. trifluoromethyl), 3 to 7 membered carbocycle group or halogenated 3- to 7-membered carbocyclyl.
• Xi is N.
• X 1 is C(R 4 ); wherein, R 4 is hydrogen, halogen, hydroxyl, cyano, -C 2-4 alkenylene-phenyl, -C 2-4 alkynylene- Phenyl, -S(O)-OH, -S(O) 2 -OH, -S-(C 1-6 alkyl) or R 8 ; R 8 is C 1-6 alkyl, -OC 1-6 Alkyl, -C 1-6 alkylene -OC 1-6 alkyl, -OC 1-6 alkylene -OC 1-6 alkyl, -C 0-6 alkylene -NRR', -C 0 -6 alkylene-C(O)OH, -C 0-6 alkylene-C(O)-C 1-6 alkyl, -C 0-6 alkylene-C(O)-NRR', -C 0-6 alkylene-NR-C(O)-C 1-6 alkyl, -C 0-6 al
• X 1 is C(R 4 ); wherein, R 4 is -C 0-6 alkylene-NRR', -C 0-6 alkylene-C(O)OH, -C 0 -6 alkylene-C(O)-C 1-6 alkyl, -C 0-6 alkylene-C(O)-NRR', -C 0-6 alkylene-NR-C(O) -C 1-6 alkyl, -C 0-6 alkylene-S(O) 2 -C 1-6 alkyl, -C 0-6 alkylene-S(O) 2 -NRR', -C 0-6 alkylene-NR-S(O) 2 -C 1-6 alkyl, -C 0-6 alkylene-NR-S(O) 2 -NRR', -C 0-6 alkylene-P(O)O-(C 1-6 alkyl) 2 ⁇ -C 0-6 alkylene-P(O)-(O)-(O
• X 1 is C(R 4 ); wherein, R 4 is -C 0-6 alkylene-P(O)O-(C 1-6 alkyl) 2 , -C 0-6 Alkylene-P (O)-(C 1-6 alkyl) (OC 1-6 alkyl) or -C 0-6 alkylene-P (O)-(C 1-6 alkyl) 2 ; Wherein, the C 1-6 alkyl, C 0-6 alkylene are independently unsubstituted or substituted by 1, 2, 3, 4 or 5 groups selected from S1 group; the group S1 group The group definition is the same as before.
• X 1 is C(R 4 ); wherein, R 4 is -P(O)O-(C 1-6 alkyl) 2 , -P(O)-(C 1-6 alkyl )(OC 1-6 alkyl) or -P(O)-(C 1-6 alkyl) 2 ; wherein, the C 1-6 alkyl is independently unsubstituted or replaced by 1, 2, 3, 4 or 5 groups selected from group S1 are substituted; the groups in group S1 are as defined above.
• X 1 is C(R 4 ); wherein R 4 is hydrogen, halo, or cyano.
• X 1 is C(R 4 ); wherein, R 4 is -P(O)O-(CH 3 ) 2 , -P(O)-(CH 3 )(O-CH 3 ) or -P(O)-(CH 3 ) 2 .
• X 2 is C(R 3 ); R 3 is hydrogen.
• Y is C( R4 ); R4 is hydrogen.
• Z is C( R3 ); R3 is hydrogen.
• two hydrogen atoms on the carbon atom on -CH 2 - are replaced by a cyclopropyl ring spiro ring, which means that two hydrogen atoms on the carbon atom on -CH 2 - are replaced by a divalent form of ethyl (i.e. ethylene After substitution, the ethylene group and the carbon atom jointly form a cyclopropyl group. and so on.
• the compound of formula (A) is a compound of formula (I-A);
• X 1 , Y, Z, X 2 , R 1 , R 2 , L 1 , L 2 , ring A and ring B are the same as above;
• the compound of formula (A) is a compound of formula (I-B);
• each of X 1 , Y, Z , X 2 , R 1 , R 2 , L 1 , L 2 , ring A, R 13 , and R 14 is as defined above; m0 is 1, 2, 3, 4, 5 or 6;
• the compound of formula (A) is a compound of formula (A1);
• X 1 , Y, Z, X 2 , R 1 , R 2 , L 1 , L 2 , ring A and ring B are the same as above;
• the compound of formula (A) is a compound of formula (A2);
• each of X 1 , Y, Z , X 2 , R 1 , R 2 , L 1 , L 2 , ring A, R 13 , and R 14 is as defined above; m0 is 1, 2, 3, 4, 5 or 6.
• the compound of formula (A) is a compound of formula (Ia), a compound of formula (Ib), a compound of formula (Ic), a compound of formula (Id), a compound of formula (Ie), or a compound of formula (If);
• X 1 , Y, Z, X 2 , R 1 , R 2 , L 1 , L 2 , ring A, and ring B are respectively defined as above.
• the compound of formula (A) is a compound of formula (II-A);
• R 1 , R 2 , L 1 , L 2 , ring A, ring B, and X 1 are each defined as above;
• the compound of formula (A) is a compound of formula (II-B);
• X 1 , R 1 , R 2 , L 1 , L 2 , Ring A, R 13 , R 14 are each as defined above; m0 is 1, 2, 3, 4, 5 or 6;
• the compound of formula (A) is a compound of formula (A3);
• X 1 , R 1 , R 2 , L 1 , L 2 , ring A, and ring B are each as defined above;
• the compound of formula (A) is a compound of formula (A4)
• X 1 , R 1 , R 2 , L 1 , L 2 , ring A, R 13 , and R 14 are each as defined above; m0 is 1, 2, 3, 4, 5 or 6.
• the compound of formula (A) is a compound of formula (IIa), formula (IIb), formula (IIc), formula (IId), formula (IIe) or formula (IIf);
• X 1 , R 1 , R 2 , L 1 , L 2 , ring A, and ring B are each as defined above.
• the compound of formula (A) is a compound of formula (III-A);
• L 1 , L 2 , Ring A, Ring B, and X 1 are the same as above;
• the compound of formula (A) is a compound of formula (III-B);
• X 1 , L 1 , L 2 , Ring A, R 13 , R 14 are as defined above; m0 is 1, 2, 3, 4, 5 or 6;
• the compound of formula (A) is a compound of formula (A5);
• the compound of formula (A) is a compound of formula (A6);
• X 1 , L 1 , L 2 , ring A, R 13 , and R 14 are each as defined above; m0 is 1, 2, 3, 4, 5 or 6.
• the compound of formula (A) is a compound of formula (III-1) or a compound of formula (III-2);
• L 1 , L 2 , ring A, ring B, and X 1 are each as defined above.
• the compound of formula (A) is a compound of formula (III-a), a compound of formula (III-b), a compound of formula (III-c) or a compound of formula (III-d);
• L 1 , L 2 , ring A, ring B, and X 1 are each as defined above.
• the compound of formula (A) is a compound of formula (IV-a), a compound of formula (IV-b), or a compound of formula (IV-c);
• L 1 , L 2 , ring B, and X 1 are as defined above.
• ring A is none; or ring A is selected from the group consisting of 3 to 6 membered carbocycle, 3 to 6 membered heterocycle, 5 to 6 membered heteroaryl, phenyl, said ring A is not Substituted or substituted by 1, 2 or 3 selected from OH, C 1-6 alkyl, halogen, and the remaining variables are as defined in the present invention.
• ring A is none; or ring A is selected from the group consisting of cyclobutyl ring, cyclopentyl ring, cyclohexyl ring, piperidine ring, piperazine ring, tetrahydropyrrole ring, pyrazole ring, imidazole ring ; Ring A is unsubstituted or substituted by 1, 2, 3, 4 or 5 groups selected from group S1, and the remaining variables are as defined in the present invention.
• ring A is none; or ring A is selected from the group consisting of cyclobutyl ring, cyclopentyl ring, cyclohexyl ring, piperidine ring, piperazine ring, tetrahydropyrrole ring, pyrazole ring, imidazole ring ; Ring A is unsubstituted or substituted by 1, 2, 3, 4 or 5 groups selected from S1 group; said group S group includes: hydroxyl, methyl, halogen, and the rest of the variables are as in the present invention defined.
• Ring A is None; or Ring A is selected from the group consisting of:
• Ring A is None; or Ring A is selected from the group consisting of:
• ring B is selected from the group consisting of 3 to 6 membered carbocycles, 3 to 6 membered heterocycles, said ring B is unsubstituted or replaced by 1, 2 or 3 members selected from OH, C 1- 6 alkyl, halogen substitution, and other variables are as defined in the present invention.
• Ring B is selected from the group consisting of cyclobutyl ring, cyclopentyl ring, cyclohexyl ring, piperidine ring, piperazine ring, tetrahydropyrrole ring, pyrazole ring, imidazole ring; Ring B is unsubstituted or substituted by 1, 2, 3, 4 or 5 groups selected from group S1, and the remaining variables are as defined in the present invention.
• Ring B is selected from the group consisting of: The remaining variables are as defined in the present invention
• Ring B is selected from the group consisting of:
• L 2 is -NR 11 -(CR 13 R 14 ) m1 -NR 12 -; R 11 , R 12 are independently hydrogen or methyl; each R 13 , R 14 are independently hydrogen, halogen or methyl; m1 is 1, 2, 3, 4, 5 or 6, and the remaining variables are as defined in the present invention.
• L 2 is -O-(CR 13 R 14 ) m1 -NR 12 -;
• R 12 is hydrogen or methyl;
• each R 13 , R 14 is independently hydrogen, halogen or methyl;
• m1 is 1, 2, 3, 4, 5 or 6, and the remaining variables are as defined in the present invention.
• L 2 is -NR 11 -(CR 13 R 14 ) m1 -;
• R 11 is hydrogen or methyl;
• each R 13 , R 14 is independently hydrogen, halogen or methyl;
• m1 is 1, 2, 3, 4, 5 or 6, and the remaining variables are as defined in the present invention.
• L 2 is -(CR 13 R 14 ) m1 -NR 12 -;
• R 12 is hydrogen or methyl;
• each R 13 , R 14 is independently hydrogen, halogen or methyl;
• m1 is 1, 2, 3, 4, 5 or 6, and the remaining variables are as defined in the present invention.
• L 2 is -(CR 13 R 14 ) m1 -; each R 13 , R 14 is independently hydrogen, halogen or methyl; m1 is 1, 2, 3, 4, 5 or 6, and the rest Variables are as defined herein.
• L 2 is -NR 11 -(CR 13 R 14 ) m5 -3 to 7-membered carbocycle-(CR 13 R 14 ) m6 -NR 12 -;
• R 11 and R 12 are independently hydrogen or Methyl; each R 13 and R 14 are independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4; said 3 to 7 members
• the carbocycle is unsubstituted or substituted by 1, 2, 3, 4 or 5 groups selected from group S1, the remaining variables are as defined in the present invention.
• L 2 is -O-(CR 13 R 14 ) m5 -3 to 7 membered carbocycle -(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl;
• each R 13 , R 14 are each independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4;
• the 3 to 7-membered carbocyclic ring is unsubstituted Or be substituted by 1, 2, 3, 4 or 5 groups selected from group S1, and the remaining variables are as defined in the present invention.
• L 2 is -(CR 13 R 14 ) m5 -3 to 7 membered carbocycle -(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl; each of R 13 , R 14 are each independently hydrogen, halogen or methyl; m5 is 0, 1, 2, 3 or 4; m6 is 0, 1, 2, 3 or 4; the 3 to 7-membered carbocyclic ring is unsubstituted or replaced by 1, 2, 3, 4 or 5 groups selected from group S1 are substituted.
• the 3- to 7-membered carbocycle is cyclobutyl, cyclopentyl, or cyclohexyl, with the remaining variables as defined herein.
• L 2 is -NR 11 -(CR 13 R 14 ) m5 -3 to 7-membered heterocycle-(CR 13 R 14 ) m6 -NR 12 ;
• R 11 and R 12 are independently hydrogen or methane each R 13 , R 14 are each independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4; the 3 to 7-membered hetero
• the ring is unsubstituted or substituted with 1, 2, 3, 4 or 5 groups selected from group S1, and the remaining variables are as defined in the present invention.
• L 2 is -O-(CR 13 R 14 ) m5 -3 to 7 membered heterocycle-(CR 13 R 14 ) m6 -NR 12 ;
• R 12 is hydrogen or methyl;
• each R 13 , R 14 are each independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4;
• the 3 to 7 membered heterocyclic ring is unsubstituted or Substituted by 1, 2, 3, 4 or 5 groups selected from group S1, and the remaining variables are as defined in the present invention.
• L 2 is -(CR 13 R 14 ) m5 -3 to 7 membered heterocycle -(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl;
• each R 13 , R 14 are each independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4;
• the 3 to 7-membered heterocyclic ring is unsubstituted or replaced by 1, 2, 3, 4 or 5 groups selected from group S1 are substituted, and the remaining variables are as defined in the present invention.
• L 2 is -NR 11 -(CR 13 R 14 ) m5 -NR 10 -(CR 13 R 14 ) m6 -NR 12 -;
• R 11 and R 12 are independently hydrogen or methyl; each R 13 and R 14 are each independently hydrogen, halogen or methyl;
• R 10 is hydrogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and other variables as defined herein.
• L 2 is -O-(CR 13 R 14 ) m5 -NR 10 -(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl; each R 13 , R 14 are each independently hydrogen, halogen or methyl;
• R 10 is hydrogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and the remaining variables are as defined in the present invention.
• L 2 is -(CR 13 R 14 ) m5 -NR 10 -(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl; each R 13 and R 14 are independently is hydrogen, halogen or methyl;
• R 10 is hydrogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and the remaining variables are as defined in the present invention.
• L 2 is -NR 11 -(CR 13 R 14 ) m5 -NR 10 C(O)-(CR 13 R 14 ) m6 -NR 12 -;
• R 11 and R 12 are independently hydrogen or Methyl; each R 13 and R 14 are independently hydrogen, halogen or methyl;
• R 10 is hydrogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4.
• the remaining variables are as defined in the present invention.
• L 2 is -O-(CR 13 R 14 ) m5 -NR 10 C(O)-(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl; each R 13 , R 14 are each independently hydrogen, halogen or methyl;
• R 10 is hydrogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and the rest of the variables are as described herein invention defined.
• L 2 is -(CR 13 R 14 ) m5 -NR 10 C(O)-(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl; each of R 13 , R 14 are each independently hydrogen, halogen or methyl;
• R 10 is hydrogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and the rest of the variables are as described in the present invention definition.
• L 2 is -NR 11 -(CR 13 R 14 ) m5 -C(O)NR 10 -(CR 13 R 14 ) m6 -NR 12 -;
• R 11 and R 12 are independently hydrogen or Methyl; each R 13 and R 14 are independently hydrogen, halogen or methyl;
• R 10 is hydrogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4.
• the remaining variables are as defined in the present invention.
• L 2 is -O-(CR 13 R 14 ) m5 -C(O)NR 10 -(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl; each R 13 , R 14 are each independently hydrogen, halogen or methyl;
• R 10 is hydrogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and the rest of the variables are as described herein invention defined.
• L 2 is -(CR 13 R 14 ) m5 -C(O)NR 10 -(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl; each of R 13 , R 14 are each independently hydrogen, halogen or methyl;
• R 10 is hydrogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and the rest of the variables are as described in the present invention definition.
• L 2 is -NR 11 -(CR 13 R 14 ) m5 -O-(CR 13 R 14 ) m6 -NR 12 -;
• R 11 and R 12 are independently hydrogen or methyl; each R 13 and R 14 are each independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and the remaining variables are as defined in the present invention.
• L 2 is -O-(CR 13 R 14 ) m5 -O-(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl;
• each R 13 and R 14 are independently m is hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and other variables are as defined in the present invention.
• L 2 is -(CR 13 R 14 ) m5 -O-(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl;
• each R 13 , R 14 is independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and other variables are as defined in the present invention.
• L 2 is -NR 11 -(CR 13 R 14 ) m5 -OC(O)-(CR 13 R 14 ) m6 -NR 12 -;
• R 11 and R 12 are independently hydrogen or methyl each R 13 ,
• R 14 is independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and other variables are as defined in the present invention.
• L 2 is -O-(CR 13 R 14 ) m5 -OC(O)-(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl;
• each R 13 , R 14 are each independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and the remaining variables are as defined in the present invention.
• L 2 is -(CR 13 R 14 ) m5 -OC(O)-(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl; each R 13 , R 14 are each independently hydrogen, halogen or methyl; m5 is 0, 1, 2, 3 or 4; m6 is 0, 1, 2, 3 or 4, and the remaining variables are as defined herein.
• L 2 is -NR 11 -(CR 13 R 14 ) m5 -C(O)O-(CR 13 R 14 ) m6 -NR 12 -;
• R 11 and R 12 are independently hydrogen or methane
• Each R 13 and R 14 are independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and the remaining variables are as defined in the present invention .
• L 2 is -O-(CR 13 R 14 ) m5 -C(O)O-(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl;
• each R 13 , R 14 are each independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and other variables are as defined in the present invention.
• L 2 is -(CR 13 R 14 ) m5 -C(O)O-(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl; each of R 13 , R 14 Each is independently hydrogen, halogen or methyl; m5 is 0, 1, 2, 3 or 4; m6 is 0, 1, 2, 3 or 4, and the remaining variables are as defined in the present invention.
• L 2 is -NR 11 -(CR 13 R 14 ) m5 -CH ⁇ CH-(CR 13 R 14 ) m6 -NR 12 -;
• R 11 and R 12 are independently hydrogen or methyl;
• Each R 13 and R 14 is independently hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and other variables are as defined in the present invention.
• L 2 is -O-(CR 13 R 14 ) m5 -CH ⁇ CH-(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl; each of R 13 , R 14 Each is independently hydrogen, halogen or methyl; m5 is 0, 1, 2, 3 or 4; m6 is 0, 1, 2, 3 or 4, and the remaining variables are as defined in the present invention.
• L 2 is -(CR 13 R 14 ) m5 -CH ⁇ CH-(CR 13 R 14 ) m6 -NR 12 -;
• R 12 is hydrogen or methyl;
• each R 13 and R 14 are independently m is hydrogen, halogen or methyl;
• m5 is 0, 1, 2, 3 or 4;
• m6 is 0, 1, 2, 3 or 4, and other variables are as defined in the present invention.
• the groups of the S1 group include: oxo, hydroxy, cyano, halogen, methyl, ethyl, propyl, and other variables are as defined in the present invention.
• the 3- to 7-membered carbocycle is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, with the remaining variables as defined herein.
• the 3- to 7-membered heterocyclic ring is azetidinyl, oxetanyl, tetrahydropyrrolyl, tetrahydrofuranyl, piperidinyl, or piperazinyl, and the remaining variables are as defined herein .
• L is selected from one of the following groups:
• L is selected from the group consisting of: The remaining variables are as defined herein.
• L 1 is -N(CD 3 )-, -CH(CD 3 ), -N(CD 3 )-C(O)- or -C(O)-N(CD 3 ), and the rest Variables are as defined herein.
• X 1 is N or C(R 4 ); wherein, R 4 is hydrogen, halogen, cyano, -S(O) 2 CH 3 or -P(O)(CH 3 ) 2 , and the rest Variables are as defined herein.
• the compound of formula (A) is selected from one of the compounds in Table (I) below:
• the second aspect of the present invention provides a pharmaceutical composition, which comprises the compound described in the first aspect, its stereoisomer, prodrug or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
• active substance of the present invention or “compound of the present invention” refers to the compound described in the first aspect, its stereoisomer, prodrug or a pharmaceutically acceptable salt thereof.
• the term "pharmaceutically acceptable carrier” refers to any formulation or carrier medium capable of delivering an effective amount of the active substance of the present invention, which does not interfere with the biological activity of the active substance and has no toxic side effects on the host or the subject.
• Carriers include water, oils, vegetables and minerals, cream bases, lotion bases, ointment bases, etc. These bases include suspending agents, viscosity builders, skin penetration enhancers and the like. Their formulations are well known to those skilled in the field of cosmetics or topical medicine.
• the pharmaceutical composition can be administered in any of the following ways: oral administration, spray inhalation, rectal administration, nasal administration, buccal administration, topical administration, parenteral administration, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, cardiac Indoor, intrasternal and intracranial injection or infusion, or with the aid of an explanted reservoir. Among them, oral, intraperitoneal or intravenous administration is preferred.
• the compounds of the present invention may be prepared in any orally acceptable preparation form, including but not limited to tablets, capsules, aqueous solutions or aqueous suspensions. Carriers used for tablets generally include lactose and corn starch, and lubricating agents such as magnesium stearate may also be added.
• Diluents used in capsule formulations generally include lactose and dried cornstarch.
• Aqueous suspensions are usually prepared by mixing the active ingredient with suitable emulsifying and suspending agents. If desired, some sweetening, flavoring or coloring agents may also be added to the above oral preparation forms.
• the compound of the present invention When using locally, especially when treating the affected areas or organs that are easily accessible by local external application, such as eyes, skin or lower intestinal nerve diseases, the compound of the present invention can be made into different topical preparations according to different affected areas or organs
• the compounds of the present invention may be formulated in the form of a micronized suspension or solution in the form of isotonic sterile saline at a certain pH with or without the addition of preservatives such as Benzyl alkoxide chloride.
• the compounds may also be formulated in ointments such as petrolatum.
• the compounds of the invention When applied topically to the skin, the compounds of the invention may be formulated in suitable ointments, lotions or creams wherein the active ingredients are suspended or dissolved in one or more carriers.
• Carriers that can be used in ointment formulations include, but are not limited to: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide, emulsifying wax, and water; carriers that can be used in lotions or creams include, but are not limited to: mineral oil Oil, sorbitan monostearate, Tween 60, cetyl esters wax, cetyl aryl alcohol, 2-octyldodecanol, benzyl alcohol and water.
• the compounds of the present invention can also be administered in the form of sterile injectable preparations, including sterile injectable aqueous or oily suspensions or sterile injectable solutions.
• Vehicles and solvents that may be employed include water, Ringer's solution, and isotonic sodium chloride solution.
• sterile, fixed oils, such as mono- or diglycerides can also be employed as a solvent or suspending medium.
• the present invention provides the use of the compound described in the first aspect, its stereoisomer, prodrug or pharmaceutically acceptable salt thereof in the preparation of a medicament for treating and/or preventing CDK7-related diseases.
• the present invention provides the use of the compound described in the first aspect, its stereoisomer, prodrug or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating and/or preventing cancer.
• the present invention provides a method of treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the compound of the first aspect, its stereoisomer, prodrug, or pharmaceutical acceptable salts above, or any combination of the above, or the step of administering the pharmaceutical composition described in the second aspect.
• the present invention provides the use of the compound described in the first aspect, its stereoisomer, prodrug or pharmaceutically acceptable salt thereof in the preparation of a CDK7 inhibitor.
• the CDK7-associated disease is a proliferative disease (eg, tumor or cancer), infectious disease, immune disease, autoimmune disease, and inflammatory disease.
• the tumor or cancer includes solid tumors, hematological tumors.
• the tumor or cancer includes melanoma, bone cancer (e.g., osteosarcoma, Ewing's sarcoma), breast cancer (e.g., hormone receptor positive (HR+) breast cancer (e.g., estrogen receptor positive (ER+) or progesterone receptor positive (PR+) breast cancer), hormone receptor negative breast cancer, triple negative breast cancer (TNBC; ER-/PR-/HER2-)), colorectal cancer, brain cancer, lung cancer (e.g.
• CLL chronic lymphocytic leukemia
• ALL acute lymphoblastic Cell leukemia
• T-ALL T-cell acute lymphoblastic leukemia
• CML chronic myeloid leukemia
• AML acute myelogenous leukemia
• lymphoma multiple myeloma
• glioblastoma Neuroblastoma.
• the infectious disease includes AIDS, cholera, conjunctivitis, dengue fever, encephalitis, enterovirus infection (e.g., polio, non-poliomyelitis), E. coli infection, foot-and-mouth disease, hepatitis, zoster Herpes, flu, measles, etc.
• enterovirus infection e.g., polio, non-poliomyelitis
• E. coli infection e.g., foot-and-mouth disease
• hepatitis hepatitis
• zoster Herpes e.g., flu, measles, etc.
• the immune and/or autoimmune diseases include asthma, diabetes, rheumatic diseases, AIDS, rejection of transplanted organs and tissues, rhinitis, chronic obstructive pulmonary disease, osteoporosis, ulcerative colitis , lupus erythematosus, allergies, rheumatoid arthritis, myasthenia gravis, Crohn's disease, psoriasis, etc.
• the inflammatory disease is selected from inflammatory diseases of the central nervous system (CNS), inflammatory rheumatic diseases, inflammatory diseases of blood vessels, inflammatory diseases of the middle ear, inflammatory bowel diseases, cutaneous Inflammatory Diseases, Uveitis Inflammatory Diseases, and Inflammatory Diseases of the Throat.
• CNS central nervous system
• inflammatory rheumatic diseases inflammatory diseases of blood vessels
• inflammatory diseases of the middle ear inflammatory diseases of the middle ear
• inflammatory bowel diseases cutaneous Inflammatory Diseases
• Uveitis Inflammatory Diseases Uveitis Inflammatory Diseases
• Inflammatory Diseases of the Throat cutaneous Inflammatory Diseases of the Throat.
• the term "subject” refers to an animal, especially a mammal, preferably a human.
• the term "effective amount” or “therapeutically effective amount” refers to a non-toxic but sufficient amount of a drug or agent to achieve the desired effect.
• the amount of a given drug depends on many factors, such as the specific dosing regimen, the type of disease or condition and its severity, the subject in need of treatment, or the uniqueness of the host (e.g. body weight), however, depending on the particular surrounding circumstances including, for example, the particular drug employed, the route of administration, the condition being treated, and the subject or host being treated, the dosage administered can be determined by those known in the art The method is routinely determined.
• the administered dosage is typically in the range of 0.02-5000 mg/day, for example about 1-1500 mg/day.
• the desired dose may conveniently be presented as one dose, or as divided doses administered simultaneously (or within a short period of time) or at appropriate intervals, for example as two, three, four or more divided doses per day.
• the specific effective dose can be adjusted appropriately according to the condition of the patient and in combination with the doctor's diagnosis.
• the term "pharmaceutically acceptable salt” refers to a salt of a compound of the present invention that is pharmaceutically acceptable and that possesses the pharmacological activity of the parent compound.
• Such salts include: acid addition salts formed with inorganic acids such as nitric acid, phosphoric acid, carbonic acid, etc.; said organic acids such as propionic acid, hexanoic acid, cypionic acid, Glycolic acid, pyruvic acid, gluconic acid, stearic acid, muconic acid, etc.; or salts formed when the acidic protons present on the parent compound are replaced by metal ions, such as alkali metal ions or alkaline earth metal ions; or with organic bases Formed coordination compound, the organic base such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine and the like.
• the pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing acid groups or bases by conventional chemical methods.
• such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.
• non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred.
• the compounds provided herein also exist in prodrug forms. Prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to convert them to the compounds of the present invention.
• prodrugs can be converted to the compounds of the invention by chemical or biochemical methods in an in vivo environment.
• stereoisomer includes conformational isomers and configurational isomers, wherein configurational isomers mainly include cis-trans isomers and optical isomers.
• the compounds of the present invention may exist in the form of stereoisomers, and thus encompass all possible stereoisomeric forms, including but not limited to cis-trans isomers, tautomers, enantiomers, non- Enantiomers, atropisomers, etc., the compounds of the present invention can also be any combination or any mixture of the aforementioned stereoisomers, such as mesoforms, racemates, atropisomers It exists in the form of a mixture of equal amounts.
• a single enantiomer, a single diastereoisomer or a mixture thereof, or a single atropisomer or a mixture thereof When the compounds described herein contain olefinic double bonds, unless otherwise specified, they include cis-isomers and trans-isomers, and any combination thereof.
• Atropisomers of the present invention are stereoisomers based on axial or planar chirality resulting from restricted intramolecular rotation.
• the compound of formula (A) has optical isomers derived from asymmetric carbon, axial asymmetry, etc., and if necessary, a single isomer can be resolved by methods known in the art, such as crystallization or chiral chromatography. .
• hydrocarbyl refers to a straight or branched chain saturated aliphatic hydrocarbyl group.
• C 1-6 hydrocarbon group refers to a straight or branched chain hydrocarbon group having 1 to 6 carbon atoms. Examples include C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl and the like.
• C 1-6 alkyl refers to a straight or branched chain alkyl having 1 to 6 carbon atoms. It is preferably C 1-4 alkyl, more preferably C 1-3 alkyl. Specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-Dimethylpropyl, 2,2-Dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2 -Methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1, 3-Dimethylbutyl, 2-ethylbutyl, 2-
• alkenyl refers to an alkyl group as defined above having one or more carbon-carbon double bonds at any position in the chain
• C 2-6 alkenyl refers to an alkyl group having 2 to 6 carbons. Atoms and at least one (eg, 1 to 2) carbon-carbon double bond.
• C 2-4 alkenyl ie, an alkenyl group having 2 to 4 carbon atoms and 1 to 2 carbon-carbon double bonds
• Specific examples include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl, pentenyl, hexenyl, butadienyl, and the like.
• alkynyl refers to an alkyl group as defined above having one or more carbon-carbon triple bonds at any position in the chain
• C alkynyl refers to an alkyl group having 2 to 6 carbons. Atoms and at least one (eg, 1 to 2) carbon-carbon triple bond. More preferred is C 2-4 alkynyl (ie, an alkynyl group having 2 to 4 carbon atoms and 1 to 2 carbon-carbon triple bonds). Specific examples include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl, and the like.
• halogen refers to fluorine, chlorine, bromine or iodine.
• halo refers to fluoro, chloro, bromo or iodo.
• halohydrocarbyl refers to a hydrocarbyl group in which one or more (eg, 1, 2, 3, 4 or 5) hydrogen atoms are replaced by halogen, wherein hydrocarbyl is as defined above.
• haloC 1-6 alkyl refers to a haloalkyl group having 1 to 6 carbon atoms. It is preferably a halogenated C 1-4 alkyl group, more preferably a halogenated C 1-3 alkyl group.
• Specific examples include, but are not limited to, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, monobromoethyl, monofluoromethyl, Difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, etc.
• deuterated hydrocarbyl refers to a hydrocarbyl group in which one or more (eg, 1, 2, 3, 4 or 5) hydrogen atoms are replaced by deuterium atoms, wherein hydrocarbyl is as defined above.
• deuterated C 1-6 alkyl refers to a deuterated alkyl group having 1 to 6 carbon atoms. It is preferably deuterated C 1-4 alkyl, more preferably deuterated C 1-3 alkyl. Specific examples include, but are not limited to, monodeuteromethyl, dideuuteromethyl, trideuteromethyl, and the like.
• carbocyclyl and “carbocycle” are used interchangeably to refer to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon group.
• the term “3- to 14-membered carbocyclyl” or “C 3-14 carbocyclyl” refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon group having 3 to 14 carbon atoms. Examples include monocyclic carbocyclyl (such as 3 to 7 membered carbocyclyl or "C 3-7 carbocyclyl”) or polycyclic carbocyclyl (such as spiro carbocyclyl, fused carbocyclyl and bridged carbocyclyl, etc. ).
• 3 to 7 membered carbocyclyl or " C3-7 carbocyclyl” can be used interchangeably, and both refer to a saturated or partially unsaturated monocyclic cyclic hydrocarbon group having 3 to 7 ring carbon atoms. It is preferably a 3- to 6-membered monocyclic carbocyclyl, preferably a 4- to 6-membered monocyclic carbocyclyl, and more preferably a 3, 4, 5, or 6-membered monocyclic carbocyclyl.
• monocyclic carbocyclyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
• spirocarbocyclyl and "spirocarbocyclyl ring” refer to a polycyclic cyclic hydrocarbon group formed by sharing one carbon atom (called a spiro atom) between two or more monocyclic rings. According to the number of spiro atoms shared between rings, spirocarbocyclyls are divided into single spirocarbocyclyls, double spirocarbocyclyls and polyspirocarbocyclyls.
• 6- to 12-membered spirocarbocyclyl refers to a polycyclic cyclic hydrocarbon group having 6 to 12 ring carbon atoms, wherein the monocyclic ring sharing a spiro atom is a 3 to 7-membered monocyclic carbocyclic ring. It is preferably a 7- to 12-membered spirocarbocyclyl, more preferably a 7- to 12-membered monospirocarbocyclyl.
• fused carbocyclyl and “fused carbocyclyl ring” refer to a polycyclic cyclic hydrocarbon group formed by two or more monocyclic rings sharing adjacent pairs of carbon atoms. According to the number of rings formed, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic condensed carbocyclic groups.
• the term “5 to 12 membered fused carbocyclyl” refers to a polycyclic cyclic hydrocarbon group having 5 to 12 ring carbon atoms, wherein the monocyclic rings sharing adjacent pairs of carbon atoms are 3 to 7 membered monocyclic carbocyclic rings. It is preferably a 7- to 12-membered condensed carbocyclic group.
• fused carbocyclyls include, but are not limited to:
• fused carbocyclic groups can be attached to the rest of the molecule through any ring atom.
• bridged carbocyclyl and “bridged carbocyclyl ring” refer to a polycyclic cyclic hydrocarbon group formed by two or more monocyclic rings sharing two carbon atoms that are not directly connected. According to the number of rings formed, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged carbocyclic groups.
• the term “5- to 12-membered bridged carbocyclyl” refers to a polycyclic cyclic hydrocarbon group having 5 to 12 ring carbon atoms, wherein any two rings share two carbon atoms that are not directly connected. It is preferably a 7- to 12-membered bridged carbocyclyl. Specific examples of bridged carbocyclyls include, but are not limited to:
• bridging carbocyclyls can be attached to the rest of the molecule through any ring atom.
• the carbocyclyl ring may be fused to an aryl, heteroaryl or heterocyclyl ring where the ring bonded to the parent structure is a carbocyclyl ring, non-limiting examples include indanyl, tetrahydronaphthalene base, benzocycloheptyl, etc.
• the above-mentioned various carbocyclic groups may be optionally substituted, and when substituted, the substituents are preferably one or more substituent groups described in the present application.
• heterocyclyl and “heterocycle” are used interchangeably and refer to saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon groups, including, for example, monocyclic heterocyclyl, spiroheterocyclyl, fused Heterocyclyl and Bridged Heterocyclyl.
• the ring carbon atoms of the heterocyclic group in the present invention may be optionally substituted by 1, 2 or 3 oxo groups to form a cyclic ketone, cyclic lactone or cyclic lactam structure.
• the 3- to 14-membered heterocyclic groups in the present invention include monocyclic heterocyclic groups (such as 3- to 7-membered heterocyclic groups), spiro heterocyclic groups, fused heterocyclic groups and bridged heterocyclic groups.
• Preferred are 4 to 7 membered monocyclic heterocyclic groups having 4 to 7 ring atoms, of which 1 or 2 ring atoms are heteroatoms. More preferred are 5 or 6 membered monocyclic heterocyclyl groups having 5 or 6 ring atoms, of which 1 or 2 ring atoms are heteroatoms.
• the heteroatom when the heteroatom is a nitrogen atom, the nitrogen atom may be substituted or unsubstituted (ie, N or NR, R being hydrogen or other substituents already defined herein).
• the ring carbon atoms of the monocyclic heterocyclic group may be optionally substituted by 1, 2 or 3 oxo groups to form a cyclic ketone, cyclic lactone or cyclic lactam structure.
• monocyclic heterocyclyl groups include, but are not limited to, aziridine, oxirane, azetidine, azetidin-2-one, oxazolidine, pyrrolidin-2-one, pyrrole Alkane-2,5-dione, 1,3-dioxolane, dihydrofuran-2(3H)-one, dihydrofuran-2,5-dione, piperidin-2-one, piperidine- 2,6-Diketone, Tetrahydro-2H-pyran-2-one, Imidazolidine, Tetrahydrofuran, Tetrahydrothiophene, Tetrahydropyrrole, 1,3-dioxolane-2-one, Oxazolidine-2 -ketone, imidazolidin-2-one, piperidine, piperazine, piperazin-2-one, morpholine, morpholin-3-one, morpholin-2-one, thiomorpholin-3-one1, 1-dioxide, thi
• the two ring atoms connected to the above-mentioned monocyclic heterocyclyl ring can optionally be combined with the monocyclic carbocyclyl ring, monocyclic heterocyclyl ring, monoaryl ring, 5 Or a carbocyclyl, heterocyclyl, aryl or heteroaryl such as a 6-membered monocyclic heteroaryl ring is fused to form a condensed polycyclic ring.
• the 2 ring atoms attached to the monocyclic heterocyclic group forming a fused ring with other rings are preferably C-C.
• the heteroatom is a nitrogen atom
• the nitrogen atom may be substituted or unsubstituted (ie, N or NR, R being hydrogen or other substituents already defined herein).
• Each single ring can contain one or more double bonds, but no ring has a fully conjugated ⁇ -electron system.
• spiroheterocyclyls are classified as single spiroheterocyclyls, double spiroheterocyclyls or polyspiroheterocyclyls.
• the term "7 to 14 membered spiroheterocyclyl” refers to a spiroheterocyclyl having 7 to 14 ring atoms, of which 1 or 2 ring atoms are heteroatoms.
• spiroheterocyclyls may be attached to the remainder of the molecule through any suitable ring atom.
• the heteroatom is a nitrogen atom
• the nitrogen atom may be substituted or unsubstituted (ie, N or NR, R being hydrogen or other substituents already defined herein).
• Each single ring can contain one or more double bonds, but no ring has a fully conjugated ⁇ -electron system.
• Shared adjacent pairs of ring atoms may be CC or NC. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic condensed heterocyclic groups.
• the term "5 to 14 membered fused heterocyclic group” refers to a condensed heterocyclic group having 5 to 14 ring atoms.
• Preferred is a 6- to 10-membered condensed heterocyclic group having 6 to 10 ring atoms, of which 1 or 2 ring atoms are heteroatoms.
• 8 to 10 membered fused heterocyclic groups having 8 to 10 ring atoms, of which 1 or 2 ring atoms are heteroatoms. Most preferred are 8-membered (5-membered monocyclic heterocyclyl ring fused to 5-membered monocyclic heterocyclyl ring), 9-membered (5-membered monocyclic heterocyclyl ring fused to 6-membered monocyclic heterocyclyl ring) Or a 10-membered (6-membered monocyclic heterocyclyl ring condensed with a 6-membered monocyclic heterocyclyl ring) bicyclic condensed heterocyclic group. Specific examples of fused heterocyclic groups include, but are not limited to:
• fused heterocyclic groups may be attached to the remainder of the molecule through any suitable ring atom.
• 5- to 14-membered bridged heterocyclic group refers to a saturated or partially unsaturated polycyclic heterocyclic group having 5 to 14 ring atoms, wherein any two rings share two ring atoms that are not directly connected, and each single Rings can contain one or more double bonds, but none have a fully conjugated ⁇ -electron system. It is preferably a 5- to 10-membered bridged heterocyclic group.
• bridged heterocyclyl groups include, but are not limited to:
• bridging heterocyclyl groups may be attached to the remainder of the molecule through any suitable ring atom.
• the above-mentioned various heterocyclic groups may be optionally substituted, and when substituted, the substituents are preferably one or more substituent groups described in the present application.
• aryl As used herein, the terms “aryl”, “aryl ring” and “aryl ring” are used interchangeably to refer to an all-carbon monocyclic ring, an all-carbon non-fused polycyclic ring (rings are covalently bonded, non-fused combined) or all-carbon fused polycyclic (that is, rings that share adjacent pairs of carbon atoms) groups, at least one ring in the group is aromatic, that is, has a conjugated ⁇ -electron system.
• C 5-12 aryl refers to an aryl group having 5 to 12 ring atoms, preferably a C 6-10 aryl group.
• the C 5-12 aryl group in the present invention includes monocyclic aryl groups (such as phenyl), non-fused polycyclic aryl groups (such as biphenyl) and aromatic fused polycyclic groups.
• the aromatic fused polycyclic ring can be a polycyclic ring formed by the fusion of a single aryl ring and one or more single aryl rings groups, non-limiting examples of which include naphthyl, anthracenyl, and the like.
• the aromatic fused polycyclic ring can also be a single aryl ring (such as phenyl) fused with one or more non-aromatic rings A polycyclic group formed by synthesis, wherein the ring connected to the parent structure is an aromatic ring or a non-aromatic ring.
• the non-aromatic ring includes but is not limited to 3 to 7 membered monocyclic heterocyclyl rings (preferably 5 or 6 membered monocyclic heterocyclyl rings, the ring carbon atoms of the monocyclic heterocyclyl rings can be replaced by 1 to 2 A oxo group is substituted to form a cyclic lactam or cyclic lactone structure), a 3 to 7-membered monocyclic carbocyclyl ring (preferably a 5 or 6-membered monocyclic carbocyclyl ring, the monocyclic carbocyclyl ring of which Ring carbon atoms can be substituted by 1 or 2 oxo groups to form a ring ketone structure).
• the polycyclic group in which the above-mentioned single aryl ring is fused with one or more non-aromatic rings can be connected to other groups or parent structures through nitrogen atoms or carbon atoms, and the rings connected to the parent structure are single aryl rings or non-aromatic ring.
• heteroaryl As used herein, the terms “heteroaryl”, “heteroaryl ring” and “heteroaryl ring” are used interchangeably to refer to a single or fused ring in which ring atoms are replaced by at least one heteroatom independently selected from nitrogen, oxygen or sulfur Polycyclic (i.e. sharing adjacent pairs of ring atoms which may be CC or NC) groups in which the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quaternized.
• the heteroaryl group has 6, 10 or 14 ⁇ -electrons shared, and at least one ring in the group is aromatic.
• the heteroaryl group herein can be a monocyclic heteroaryl (eg 5 or 6 membered monocyclic heteroaryl), a fused bicyclic heteroaryl (eg 8 to 10 membered bicyclic heteroaryl) or a fused tricyclic heteroaryl .
• monocyclic heteroaryl groups include, but are not limited to, thiophene, furan, thiazole, isothiazole, imidazole, oxazole, pyrrole, pyrazole, triazole, 1,2,3-triazole, 1,2,4-triazole, Azole, 1,2,5-triazole, 1,3,4-triazole, tetrazole, isoxazole, oxadiazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole , 1,2,5-oxadiazole, 1,3,4-oxadiazole, thiadiazole, pyridine, pyridazine, pyrimidine, pyrazine, etc.
• the fused bicyclic heteroaryl group can be a bicyclic group formed by the fusion of a monoaryl ring (such as phenyl) and a monocyclic heteroaryl ring (preferably a 5- or 6-membered monocyclic heteroaryl ring), or Can be a bicyclic group formed by the fusion of a monocyclic heteroaryl ring (preferably a 5- or 6-membered monocyclic heteroaryl ring) and a monocyclic heteroaryl ring (preferably a 5- or 6-membered monocyclic heteroaryl ring) .
• any two ring atoms connected to each other on the above-mentioned monocyclic heteroaryl ring can be combined with the monocyclic carbocyclyl ring, monocyclic heterocyclyl ring, monoaryl ring, 5 Or a carbocyclyl, heterocyclyl, aryl or heteroaryl such as a 6-membered monocyclic heteroaryl ring is fused to form a condensed polycyclic ring.
• Non-limiting examples of the 8- to 10-membered bicyclic heteroaryl include: benzo[d]isoxazole, 1H-indole, isoindole, 1H-benzo[d]imidazole, benzo[d]iso Thiazole, 1H-benzo[d][1,2,3]triazole, benzo[d]oxazole, benzo[d]thiazole, indazole, benzofuran, benzo[b]thiophene, quinoline , isoquinoline, quinazoline, quinoxaline, cinnoline, pyrido[3,2-d]pyrimidine, pyrido[2,3-d]pyrimidine, pyrido[3,4-d]pyrimidine, pyridine A[4,3-d]pyrimidine, 1,8-naphthyridine, 1,7-naphthyridine, 1,6-naphthyridine, 1,5-naph
• the above-mentioned monocyclic heteroaryl group, or the bicyclic heteroaryl group formed by the fusion of the benzene ring and the monocyclic heteroaryl ring, or the bicyclic heteroaryl group formed by the fusion of the monocyclic heteroaryl ring and the monocyclic heteroaryl ring can be obtained by
• the nitrogen or carbon atoms are linked to other groups or parent structures.
• Specific examples of bicyclic heteroaryls include, but are not limited to: These groups may be attached to the remainder of the molecule through any suitable ring atom.
• the ring attached to the parent structure can be a monocyclic heteroaryl ring or a benzene ring.
• the fused bicyclic heteroaryl or fused tricyclic heteroaryl can be a monocyclic heteroaryl ring (preferably a 5- or 6-membered monocyclic heteroaryl ring) with one or more A polycyclic group formed by the fusion of non-aromatic rings, in which the ring connected to the parent structure is a monocyclic heteroaryl ring or a non-aromatic ring.
• the non-aromatic ring includes but is not limited to 3 to 6 membered monocyclic heterocyclyl rings (preferably 5 or 6 membered monocyclic heterocyclyl rings, the ring carbon atoms of the monocyclic heterocyclyl rings can be replaced by 1 to 2 A oxo group is substituted to form a cyclic lactam or cyclic lactone structure), a 3 to 6-membered monocyclic carbocyclyl ring (preferably a 5 or 6-membered monocyclic carbocyclyl ring, the monocyclic carbocyclyl ring of which Ring carbon atoms can be substituted by 1 or 2 oxo groups to form a ring ketone structure) and the like.
• the polycyclic group formed by the fusion of the above-mentioned monocyclic heteroaryl ring and one or more non-aromatic rings can be connected to other groups or the parent structure through a nitrogen atom or a carbon atom, and the ring connected to the parent structure is a monocyclic heteroaryl base ring or non-aromatic ring.
• the term "8- to 10-membered heteroarylheterocyclyl” or “8- to 10-membered heteroarylheterocyclyl ring” refers to a 5- or 6-membered monocyclic heteroaryl ring in combination with a 5- or 6-membered monocyclic heterocycle
• An 8- to 10-membered fused bicyclic heteroaryl group formed by fused base rings that is, a fused 5- or 6-membered ring atom formed by two adjacent substituent groups on a 5- or 6-membered monocyclic heteroaryl group and the ring atoms connected to it
• a membered monocyclic heterocyclyl ring, said 5 or 6 membered monocyclic heterocyclyl ring is as defined above.
• the term "8 to 10 membered heteroarylcarbocyclyl” or “8 to 10 membered heteroarylcarbocyclyl ring” refers to a 5 or 6 membered monocyclic heteroaryl ring with a 5 or 6
• Non-limiting examples of "8 to 10 membered heteroarylheterocyclyl” or “8 to 10 membered heteroarylcarbocyclyl” include: The aforementioned groups may be attached to the rest of the molecule through any suitable ring atom thereof.
• the various heteroaryl groups mentioned above may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more substituent groups described in the present application.
• hydroxyl refers to -OH.
• amino refers to -NH2 .
• cyano refers to -CN.
• nitro refers to -NO2 .
• benzyl refers to -CH2 -benzene.
• alkylene means a divalent combination of an alkyl group.
• C 0-6 alkylene means an alkylene group having 0 to 6 carbon atoms.
• C 1 alkylene means an alkylene group having 1 carbon atom, eg -CH 2 -.
• C2alkylene means an alkylene group having 2 carbon atoms, eg -CH2CH2- . and so on.
• alkenylene means a divalent bond of alkenyl.
• C 2-6 alkenylene means an alkenylene group having 2 to 6 carbon atoms.
• substituted means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence of the specific atom is normal and the substituted compound is stable.
• it means that two hydrogen atoms are replaced.
• Oxo substitution does not occur on aryl groups.
• optionally substituted or “optionally substituted” means that it may or may not be substituted, and unless otherwise specified, the type and number of substituents may be arbitrary on a chemically feasible basis.
• any variable eg, R
• its definition is independent at each occurrence.
• said group may optionally be substituted with up to two R, with independent options for each occurrence of R.
• substituents and/or variations thereof are permissible only if such combinations result in stable compounds.
• the term “none” means that the two groups it is attached to are directly connected, such as When the ring A is nothing, L 1 and L 2 are directly connected, which is “L 1 -L 2 ".
• this substituent can be bonded through any atom, for example, pyridyl as a substituent can be connected to any atom on the pyridine ring.
• the carbon atom is attached to the group being substituted.
• linking group listed does not indicate its linking direction
• its linking direction is arbitrary, for example Among them, L is selected from -C(O)-NH-, at this time -C(O)-NH- can be connected to benzene ring and cyclopentane in the same direction as the reading order from left to right to form The benzene ring and cyclopentane can also be joined in the opposite direction to the left-to-right reading order to form
• Combinations of the described linking groups, substituents and/or variations thereof are permissible only if such combinations result in stable compounds.
• the compound represented by formula (A) of the present invention can be prepared by using a synthetic method known in the art or by combining a method known in the art with the method described in the present invention. Solvents, temperatures and other reaction conditions given in the present invention are exemplary and can be varied according to methods well known in the art.
• the example compounds described in the present invention can be synthesized according to the methods described in the examples using appropriate starting materials according to their specific structures, or can be synthesized using a method similar to that described in the examples.
• the starting materials used to synthesize the compounds of the examples of the present invention can be prepared by known synthetic methods or similar methods described in literature or obtained from commercial sources.
• the compound can be further resolved to obtain its stereoisomers by methods well known in the art, such as crystallization, chromatography, etc., if necessary, and the resolution conditions are easily obtained by those skilled in the art through routine means or limited experiments.
• the compound of formula (A) of the present invention can be synthesized by the following method, wherein the solvent, temperature and other reaction conditions in each step can be the same or similar to those described in the following examples, or use known methods in the art reaction conditions.
• the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and the methods well known to those skilled in the art equivalent replacement.
• Preferred embodiments include, but are not limited to, the examples of the present invention.
• Step 1 Add [2-(2-hydroxyethoxy)ethyl]benzyl carbamate (5.0g, 20.92mmol) and dichloromethane (25mL) into a three-necked flask, stir and add triethylamine (6.34g, 62.76mmol) and 4-dimethylaminopyridine (510mg, 4.18mmol). The mixture was cooled to 0 °C and p-toluenesulfonyl chloride (3.97 g, 20.92 mmol) was added slowly. The reaction was stirred at room temperature for 2 hours, and the reaction of the raw materials was monitored by LC-MS to complete.
• Step 2 2-(2-(((Benzyloxy)carbonyl)amino)ethoxy)ethyl 4-methylbenzenesulfonate (1000mg, 2.542mmol) and (S)-pyrrolidin-3-yl
• potassium iodide 0.056mL, 0.508mmol
• potassium carbonate 7.02mg, 5.083mmol
• Step 3 Benzyl (S)-(2-(2-(3-((tert-butoxycarbonyl)amino)pyrrolidin-1-yl)ethoxy)ethyl)carbamate (730mg, 1.791 mmol) in dichloromethane (6.0 mL), was added trifluoroacetic acid (4.0 mL, 53.85 mmol). The reaction mixture was stirred at room temperature for 3 hours, and the reaction was complete. The reaction solution was concentrated and extracted by adding ethyl acetate (50mLX3) and saturated sodium bicarbonate (50mLX3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated.
• Step 4 At room temperature, aluminum trichloride (12.9 g, 85.62 mmol). Raise the temperature to 90°C and react for 30 minutes. A solution of indole 6-methylcarboxylate (5.0 g, 28.54 mmol) in 1,2-dichloroethane (30.0 mL) was slowly added dropwise to the reaction solution. The reaction mixture was continued at 90°C for 18 hours. Cooled and poured into ice water (200 mL), and extracted with ethyl acetate (100 mL ⁇ 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated.
• indole 6-methylcarboxylate 5.0 g, 28.54 mmol
• 1,2-dichloroethane 30.0 mL
• Step 5 Under nitrogen protection at room temperature, compound 3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (450mg, 1.27mmol) and compound To a solution of benzyl (S)-(2-(2-(3-aminopyrrolidin-1-yl)ethoxy)ethyl)carbamate (583 mg, 1.9 mmol) in dioxane (6 mL) was added N , N-Diisopropylethylamine (1.56ml, 9.4mmol). The reaction mixture was warmed up to 120°C for 4 hours.
• Step 6 At room temperature, the compound methyl (R)-3-(2-((1-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)ethyl)pyrrolidine-3 10% palladium on carbon ( 100mg, 0.945mmol). The reaction mixture was stirred for about 4 hours under hydrogen atmosphere.
• Step 7 Under nitrogen protection at room temperature, the compound (R)-3-(2-((1-(2-(2-aminoethoxy)ethyl)pyrrolidin-3-yl)amino)-5-(tri)
• methanol/water (10/5mL) methanol/water (10/5mL)
• lithium hydroxide (197.3mg, 4.7mmol) was added. The reaction mixture was heated to 70°C for 3 hours. Cool and concentrate.
• Step 8 Under the protection of nitrogen at room temperature, the N , N,N-Diisopropylethylamine (0.11ml, 0.66mmol) was added to a solution of N-dimethylformamide (15mL).
• Compound (R)-3-(2-((1-(2-(2-aminoethoxy)ethyl)pyrrolidin-3-yl)amino)-5-(trifluoromethyl) was added under stirring the mixture
• Step 1 Add di-tert-butyl dicarbonate (3.67 g, 16.856 mmol) to a solution of 4-amino-1-butanol (3.0 g, 33.7 mmol) in ethanol (30 mL) at room temperature. The reaction mixture was stirred at room temperature for 4 hours. The reaction solution was concentrated, the crude product was extracted with ethyl acetate (100 mL*3) and sodium bicarbonate (150 mL), the organic phases were combined, dried over anhydrous sodium sulfate and concentrated.
• the crude product was purified by silica gel column (ethyl acetate/petroleum ether: 0-100%) to obtain tert-butyl (4-hydroxybutyl)carbamate (5.8 g, 30.69 mmol, yield: 91%) as a colorless oily product.
• Step 2 Under nitrogen protection, add oxalyl chloride (13ml, 23mmol, 2M dichloromethane) dropwise to the reaction solution of dimethyl sulfoxide (3.7ml) in dichloromethane (50ml) at -70°C, and continue Stir for 5 minutes. A solution of tert-butyl (4-hydroxybutyl)carbamate (4.1 g, 21.6 mmol) in dichloromethane (35.0 mL) was added dropwise at -70°C. The reaction mixture was gradually warmed to 0°C and stirred for 25 minutes. After the reaction was completed, it was poured into ice water, and extracted with ethyl acetate (50mLX3).
• oxalyl chloride 13ml, 23mmol, 2M dichloromethane
• Step 3 Under the protection of nitrogen at room temperature, compound 3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (2.0g, 5.62mmol) and Add N,N-diisopropylethylamine ( 2.78ml, 16.78mmol). The reaction mixture was heated to 140°C for 0.5 hours. Cooled and poured into ice water, and extracted with ethyl acetate (50mLX3). The organic phases were combined, washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, and concentrated.
• Step 4 Compound 3-(2-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidine-4- at room temperature
• dichloromethane 10 mL
• trifluoroacetic acid 1.0 mL, 13.56 mmol
• the reaction solution was concentrated and extracted by adding ethyl acetate (50mL ⁇ 3) and saturated sodium bicarbonate (100mL).
• Step 5 Compound methyl 3-(2-(((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole at room temperature
• -6-carboxylate 480 mg, 1.0 mmol
• sodium cyanoborohydride 143.84 mg, 2.29 mmol
• reaction solution was concentrated and extracted by adding ethyl acetate (50mL ⁇ 3) and saturated sodium bicarbonate (100mL). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated.
• Step 6 Compound methyl 3-(2-(((1S,3S)-3-((4-((tert-butoxycarbonyl)amino)butyl)amino)cyclopentyl)amino)-5- at room temperature (Trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylate (356mg, 0.76mmol) in dichloromethane (5mL) solution, add trifluoroacetic acid (0.5mL, 6.75mmol) . The reaction mixture was stirred at room temperature for 1 hour, and the reaction was complete. The reaction solution was concentrated and extracted by adding ethyl acetate (50mL ⁇ 3) and saturated sodium bicarbonate (100mL).
• Step 7 Under nitrogen protection at room temperature, the compound 3-(2-(((1S,3S)-3-((4-aminobutyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidine
• methyl-4-yl)-1H-indole-6-carboxylate 384 mg, 0.78 mmol
• lithium hydroxide 98.54 mg, 2.35 mmol
• Step 8 Under the protection of nitrogen at room temperature, the N , N,N-Diisopropylethylamine (0.139ml, 0.84mmol) was added to the solution of N-dimethylformamide (5mL).
• Step 1 Under nitrogen protection, add 3-N-tert-butoxycarbonyl-6-oxa-3-azabicyclo[3.1.0]hexane (5g, 26.99mmol) and cuprous bromide at -30°C (1) Allylmagnesium chloride (60.739mL, 121.48mmol, 2M/L) was slowly added dropwise in a tetrahydrofuran (150mL) solution of dimethyl sulfide complex (60.739mL, 121.48mmol), and the reaction solution was under nitrogen protection. The reaction was stirred at -10°C for 1 hour.
• reaction solution was quenched with 10% aqueous ammonium chloride solution (100 mL) and ethyl acetate (100 mL), and then stirred at room temperature for 1 hr.
• the reaction solution was extracted with ethyl acetate (50 mL ⁇ 3), and the combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness.
• Step 2 Under the protection of nitrogen, add (3R,4S)-3-hydroxy-4-allylpyrrolidine-1-carboxylic acid tert-butyl ester (1.5g, 6.60mmol) to N,N-di Sodium hydride (0.32 g, 7.92 mmol, 60% purity) was added to the methylformamide (8 mL) solution, and the reaction solution was stirred and reacted at room temperature for 0.5 hours under the protection of nitrogen. Benzyl bromide (0.947 mL, 7.92 mmol) was added thereto, and the resulting reaction solution was stirred overnight at room temperature.
• Step 3 To (3R,4S)-3-benzyloxy-4-allylpyrrolidine-1-carboxylic acid tert-butyl ester (800mg, 2.52mmol) and sodium periodate (1.62g, 7.56mmol) Potassium osmate (92.86 mg, 0.25 mmol) was added to a mixed solution of acetonitrile (15 mL) and water (6 mL), and the reaction solution was stirred at room temperature for 2 hours. Water (10 mL) was added to the reaction solution and extracted with ethyl acetate (10 mL ⁇ 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness.
• Step 4 Add (3R,4S)-3-benzyloxy-4-(2-acetaldehyde)pyrrolidine-1-carboxylic acid tert-butyl ester (480mg, 1.50mmol) and 3-(2-(( (1S,3S)-3-amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (630mg, 1.50mmol) , A small amount of acetic acid (0.086 mL, 1.50 mmol) was added to a solution of 2-dichloroethane (15 mL), and the reaction solution was stirred at room temperature for 0.5 hours.
• Step five to 3-(2-(((1S,3S)-3-((2-((3S,4R)-4-(benzyloxy)-1-(tert-butyloxycarbonyl)pyrrolyl- 3-yl)ethyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (480mg, 0.66mmol) in tetrahydrofuran ( 2 mL) and methanol (2 mL) was added a solution of lithium hydroxide (139 mg, 3.32 mmol) in water (2 mL), and the reaction solution was stirred at room temperature overnight.
• Step 6 3-(2-(((1S,3S)-3-((2-((3S,4R)-4-(benzyloxy)-1-(tert-butyloxycarbonyl) )pyrrolyl-3-yl)ethyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (390mg, 0.55mmol) A hydrochloric acid/dioxane (10 mL, 4N) solution was added, and the reaction solution was stirred at room temperature for 1 hour.
• Step 7 To 3-(2-(((1S,3S)-3-((2-((3S,4R)-4-(benzyloxy)pyrrolyl-3-yl)ethyl)amino) ring Pentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (320mg, 0.53mmol) in N,N-dimethylformamide (180mL) solution Pentafluorophenol (194mg, 1.05mmol) and N,N'-dicyclohexylcarbodiimide (217mg, 1.05mmol) were added sequentially, and the reaction solution was stirred overnight at room temperature.
• Step 8 To (4 1 S,4 3 S,8 3 S,8 4 R)-8 4 -(benzyloxy)-2 5 -(trifluoromethyl)-1 1 H-3 at 0°C ,5-diaza-1(3,6)-indole-2(4,2)-pyrimidine-8(3,1)-pyrrolidine-4(1,3)-cyclopentanecyclononane- A solution of 9-ketone (30mg, 0.05mmol) in dichloromethane (2mL) was slowly added dropwise with a solution of boron trichloride (0.25mL, 1M), and the reaction solution was stirred at 0°C for 1.5 hours under nitrogen protection.
• 9-ketone (30mg, 0.05mmol) in dichloromethane (2mL) was slowly added dropwise with a solution of boron trichloride (0.25mL, 1M), and the reaction solution was stirred at 0°C for 1.5 hours under nitrogen protection.
• Step 1 Add N,N-diisopropylethylamine ( 2.61g, 20.0mmol) and 2,4-dichloro-5-(trifluoromethyl)pyrimidine (1.10g, 5.0mmol), the reaction solution was stirred at 25°C for 14 hours. After the completion of the reaction detected by LCMS, the reaction solution was directly concentrated to obtain the crude product.
• Step 5 To 1-(tert-butyl)6-methyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole -1,6-Dicarboxylate (2.3g, 5.73mmol) in dioxane and aqueous solution (23mL) were added potassium carbonate (1.58g, 11.46mmol), palladium dichloride (0.28g, 0.38 mmol) and tert-butyl N-[(1S,3S)-3- ⁇ [4-chloro-5-(trifluoromethyl)pyrimidin-2-yl]amino ⁇ cyclopentyl]carbamate (1.46g, 3.82 mmol), the reaction was stirred at 100°C for 3 hours.
• Step 7 To 3-(2-(((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxy Add sodium borohydride (0.39mL, 11.92mmol), tert-butyl 4-(2-oxoethyl)piperidine to a solution of methyl ester (1.00g, 2.38mmol) in 1,2-dichloroethane (10mL) -1-Carboxylate (0.54g, 2.38mmol), the reaction was stirred at room temperature overnight.
• Step 8 To methyl 3-(2-(((1S,3S)-3-((2-(1-(tert-butoxycarbonyl)piperidin-4-yl)ethyl)amino)cyclopentyl) Amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylate (150 mg, 0.24 mmol) in THF/methanol/water (1:1:1) Lithium hydroxide (0.02 mL, 0.71 mmol) was added and the reaction was stirred at room temperature overnight. The pH of the reaction was adjusted to acidic with dilute hydrochloric acid.
• Step 10 To 3-(2-((1S,3S)-3-((2-(piperidin-4-yl)ethyl)amino)cyclopentylamino)-5-(trifluoromethyl)pyrimidine -4-yl)-1H-indole-6-carboxylic acid (90mg, 0.17mmol) in N,N-dimethylformamide (5mL) solution was added 2-(7-azabenzotriazole )-N,N,N',N'-tetramethyluronium hexafluorophosphate (72mg, 0.19mmol) and N,N-diisopropylethylamine (88mg, 0.68mmol), the reaction solution was stirred at room temperature for 16 Hour.
• Step 1 At room temperature, to 3-(2-(((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(di Methylphosphoryl)-1H-indole-6-carboxylate (270mg, 0.55mmol) in N,N-dimethylformamide solution (5mL) was successively added tert-butyl (4-bromobutyl) Carbamate (138mg, 0.55mmol) and N,N-diisopropylethylamine (284mg, 2.20mmol), the reaction solution was stirred at 80°C for 16 hours under the protection of nitrogen.
• reaction solution was added with 10% aqueous ammonium chloride (100 mL) and extracted with ethyl acetate (100 mL ⁇ 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated.
• Step 2 To methyl 3-(2-(((1S,3S)-3-((4-((tert-butoxycarbonyl)amino)butyl)amino)cyclopentyl)amino)-5-(three Mixed solution of fluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (170mg, 0.26mmol) in tetrahydrofuran (2mL) and methanol (2mL) A solution of lithium hydroxide (66.0mg, 1.56mmol) in water (2mL) was added, and the reaction solution was stirred at 40°C overnight.
• Step 3 3-(2-(((1S,3S)-3-((4-((tert-butoxycarbonyl)amino)butyl)amino)cyclopentyl)amino)-5- (Trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid (160 mg, 0.25 mmol) in methanol (5 mL) was added hydrochloric acid/methanol ( 5mL, 4N) solution, the reaction solution was stirred and reacted at 25°C for 16 hours.
• Step 4 To 3-(2-(((1S,3S)-3-((4-aminobutyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl) 2-(7-aza Benzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (91.2mg, 0.24mmol) and N,N-diisopropylethylamine (114mg, 0.88mmol), The reaction solution was stirred at 80°C for 16 hours.
• Step 1 Add vinylmagnesium bromide in tetrahydrofuran (1M, 163mL, 163mmol) dropwise slowly to a solution of 2-bromo-3-nitrobenzoic acid (10.0g, 40.6mmol) in tetrahydrofuran (150mL) at -78°C ), after the dropwise addition, the reaction was continued to stir at -78°C for 3 hours under the protection of nitrogen, and then naturally rose to room temperature and stirred for 16 hours.
• Step 2 Add iodomethane (6.66g, 46.9 mmol) and potassium carbonate (10.8g, 78.3mmol). The reaction solution was stirred at 25° C. for 16 hours under nitrogen protection. After the reaction was complete as detected by LCMS, the reaction solution was added with water (200 mL), and then extracted with ethyl acetate (200 mL ⁇ 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated.
• Step 3 At 25°C, add tris(dibenzylideneacetone)dipalladium (242mg, 0.393mmol), 4,5-bisdiphenylphosphine-9,9-dimethyloxa Anthracene (455mg, 0.787mmol) and 1,4-dioxane (20mL) solution was stirred for 30 minutes under nitrogen protection, and then 7-bromo-1H-indole-6-carboxylic acid methyl ester (2.0 g, 7.87mmol), potassium phosphate (1.67g, 7.87mmol) and dimethylphosphine oxide (0.61g, 7.87mmol). The reaction solution was stirred at 110°C for 14 hours.
• Step 4 In a nitrogen atmosphere, in a dry 50mL three-necked flask, add aluminum trichloride (1.06g, 7.96mmol), and then add 7-(dimethylphosphoryl)-1H-indole-6- A solution of methyl carboxylate (1.00g, 3.98mmol) in 1.2-dichloroethane (10mL) was stirred at 80°C for 30 minutes, then 2,4-dichloro-5-(trifluoromethyl)pyrimidine (0.95 g, 4.38mmol) in 1.2-dichloroethane (1mL), the reaction solution was reacted at 80°C for 16 hours.
• Step 5 In a 10mL microwave tube, sequentially add 3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6- Methyl carboxylate (770mg, 1.78mmol), N-methylpyrrolidone (4mL), N,N-diisopropylethylamine (0.59mL, 3.57mmol) and tert-butyl ((1S,3S)-3- Aminocyclopentyl) carbamate (392mg, 1.96mmol), the reaction solution was stirred at 80°C to dissolve, and then microwaved at 130°C for 2 hours.
• Step 6 At room temperature, methyl 3-(2-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidine- 4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (400 mg, 0.67 mmol) dissolved in hydrogen chloride/dioxane (5 mL, 4M) and methanol (2 mL) The mixed solution was reacted at 20°C for 16 hours.
• Step 7 3-(2-(((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl )-1H-indole-6-carboxylic acid methyl ester (500mg, 1.0mmol) was dissolved in N,N-dimethylformamide (3mL), tert-butyl (5-bromopentyl) carbamate ( 269 mg, 1.0 mmol) and N,N-diisopropylethylamine (0.67 mL, 4.0 mmol) were added sequentially and the reaction was stirred at 80°C for 16 hours.
• Step 8 Methyl 3-(2-(((1S,3S)-3-((5-((tert-butoxycarbonyl)amino)pentyl)amino)cyclopentyl)amino)-5-(tri Fluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (360 mg, 0.53 mmol) dissolved in water (1 mL), methanol (1 mL) and THF (1 mL), then add lithium hydroxide monohydrate (111 mg, 2.65 mmol), and stir at 40°C for 16 hours.
• Step 9 3-(2-(((1S,3S)-3-((5-((tert-butoxycarbonyl)amino)pentyl)amino)cyclopentyl)amino)-5-(trifluoromethyl Base) pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid (320mg, 0.47mmol) was dissolved in hydrogen chloride/methanol (5mL, 4M), and the system was heated at 25°C Stir for 4 hours.
• Step ten to 3-(2-(((1S,3S)-3-((5-aminopentyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl) 2-(7-aza Benzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (134mg, 0.35mmol) and N,N-diisopropylethylamine (0.23mL, 1.41mmol), The reaction solution was stirred at 60°C for 16 hours.
• Step 1 At room temperature, to 3-(2-(((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(di (2-(2-Bromoethoxy ) ethyl) tert-butyl carbamate (270mg, 1.00mmol) and N,N-diisopropylethylamine (0.67mL, 4.00mmol), the reaction solution was stirred at 70°C for 16 hours under nitrogen protection.
• reaction solution was added with 10% aqueous ammonium chloride (100 mL), extracted with ethyl acetate (100 mL ⁇ 3), the combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated.
• Step 2 To methyl 3-(2-(((1S,3S)-3-((2-(2-((tert-butoxycarbonyl)amino)ethoxy)ethyl)amino)cyclopentyl) Amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (350mg, 0.51mmol) in tetrahydrofuran (2mL) and A solution of lithium hydroxide (86.0 mg, 2.05 mmol) in water (2 mL) was added to a mixed solution of methanol (2 mL), and the reaction solution was stirred at 40° C. for 16 hours.
• Step 3 3-(2-(((1S,3S)-3-((2-(2-((tert-butoxycarbonyl)amino)ethoxy)ethyl)amino) ring at 25°C Pentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid (300mg, 0.45mmol) in methanol (5mL ) solution was added hydrochloric acid/methanol (5 mL, 4M) solution, and the reaction solution was stirred at 40° C. for 4 hours.
• Step 4 To 3-(2-(((1S,3S)-3-((2-(2-aminoethoxy)ethyl)amino)cyclopentyl)amino)-5-(trifluoromethyl )pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid (220mg, 0.39mmol) in N,N-dimethylformamide (8mL) solution was added successively 2-(7-Azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (149mg, 0.39mmol) and N,N-diisopropylethylamine ( 182mg, 1.41mmol), the reaction solution was stirred at 60°C for 16 hours.
• Step 1 In a 10mL microwave tube, sequentially add 3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6- Methyl carboxylate (810mg, 1.88mmol), N-methylpyrrolidone (5mL), N,N-diisopropylethylamine (0.93mL, 5.63mmol) and tert-butyl ((1r,3r)-3- Aminocyclobutyl) carbamate (384mg, 2.06mmol), the reaction solution was stirred at 80°C to dissolve, and then microwaved at 130°C for 2 hours.
• Step 2 At room temperature, methyl 3-(2-(((1r,3r)-3-((tert-butoxycarbonyl)amino)cyclobutyl)amino)-5-(trifluoromethyl)pyrimidine- 4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (500 mg, 0.86 mmol) dissolved in hydrogen chloride/dioxane (5 mL, 4M) and methanol (2 mL) The mixed solution was reacted at 25°C for 16 hours.
• Step three 3-(2-(((1r,3r)-3-aminocyclobutyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethyl tert-Butyl (4-bromobutyl)amino Formate (157mg, 0.62mmol) and N,N-diisopropylethylamine (322mg, 2.49mmol), the reaction solution was stirred at 80°C for 16 hours under the protection of nitrogen.
• reaction solution was added to saturated ammonium chloride solution (100 mL), and extracted with ethyl acetate (100 mL ⁇ 3), the combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. .
• Step 4 To methyl 3-(2-(((1r,3r)-3-((4-((tert-butoxycarbonyl)amino)butyl)amino)cyclobutyl)amino)-5-(three Mixed solution of fluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (225 mg, 0.35 mmol) in tetrahydrofuran (2 mL) and methanol (2 mL) A solution of lithium hydroxide monohydrate (72.0 mg, 1.72 mmol) in water (2 mL) was added, and the reaction solution was stirred and reacted at 40° C. for 16 hours.
• Step five to 3-(2-(((1r,3r)-3-((4-((tert-butoxycarbonyl)amino)butyl)amino)cyclobutyl)amino)-5-(trifluoro
• methanol methanol
• hydrochloric acid/methanol 5mL, 4M
• Step 6 To 3-(2-(((1r,3r)-3-((4-aminobutyl)amino)cyclobutyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl) 2-(7-aza Benzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (118mg, 0.31mmol) and N,N-diisopropylethylamine (200mg, 1.55mmol), the The reaction solution was stirred at 70°C for 16 hours.
• Step 1 In a 10mL microwave tube, sequentially add 3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6- Methyl carboxylate (450mg, 1.04mmol), N-methylpyrrolidone (5mL), N,N-diisopropylethylamine (0.52mL, 3.13mmol) and tert-butyl ((1S,3S)-3- Aminocyclohexyl) carbamate (235mg, 1.09mmol), the reaction solution was stirred at 80°C to dissolve, and then microwaved at 130°C for 2 hours.
• Step 2 At room temperature, methyl 3-(2-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclohexyl)amino)-5-(trifluoromethyl)pyrimidine-4 -yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (470mg, 0.77mmol) was dissolved in a mixture of hydrogen chloride/dioxane (5mL, 4M) and methanol (2mL) solution, reacted at 25°C for 16 hours.
• Step 3 3-(2-(((1S,3S)-3-aminocyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl) -1H-indole-6-carboxylic acid methyl ester (310mg, 0.61mmol) was dissolved in N,N-dimethylformamide (3mL), tert-butyl (4-bromobutyl) carbamate (153mg , 0.61 mmol) and N,N-diisopropylethylamine (0.67 mL, 4.0 mmol) were added sequentially, and the reaction was stirred at 80°C for 16 hours.
• Step 4 Methyl 3-(2-(((1S,3S)-3-((4-((tert-butoxycarbonyl)amino)butyl)amino)cyclohexyl)amino)-5-(trifluoro Methyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (200 mg, 0.29 mmol) was dissolved in water (1 mL), methanol (1 mL) and tetrahydrofuran ( 1 mL), then lithium hydroxide monohydrate (61.6 mg, 1.47 mmol) was added, and the reaction was stirred at 40°C for 16 hours.
• Step five 3-(2-(((1S,3S)-3-((4-((tert-butoxycarbonyl)amino)butyl)amino)cyclohexyl)amino)-5-(trifluoromethyl )pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid (180mg, 0.27mmol) was dissolved in hydrogen chloride/methanol (5mL, 4M), and the system was stirred at 15°C 4 hours.
• Step 6 To 3-(2-(((1S,3S)-3-((4-aminobutyl)amino)cyclohexyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)- Add 2-(7-azabenzene Triazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (80.4mg, 0.21mmol) and N,N-diisopropylethylamine (0.14mL, 0.84mmol), The reaction solution was stirred at 60°C for 16 hours.
• Step 1 3-(2-(((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethyl 3-(tert-butoxycarbonylamino)propyl Bromine (144mg, 0.61mmol) and N,N-diisopropylethylamine (313mg, 2.42mmol), the reaction solution was stirred at 80°C for 16 hours under the protection of nitrogen.
• reaction solution was added with 10% aqueous ammonium chloride (50 mL), extracted with ethyl acetate (50 mL ⁇ 3), the combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated.
• Step 2 To 3-(2-(((1S,3S)-3-(3-(tert-butoxycarbonylaminopropyl)amino)cyclopentylamino)-5-(trifluoromethyl)pyrimidine-4 -yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid methyl ester (100 mg, 0.16 mmol) in tetrahydrofuran (1.5 mL) and methanol (1.5 mL) was added monohydrate Lithium hydroxide (33mg, 0.78mmol) in water (1.5mL) solution, the reaction solution was stirred and reacted at 40°C for 16 hours.
• Step 3 3-(2-(((1S,3S)-3-((3-(tert-butoxycarbonyl)amino)propyl)amino)cyclopentyl)amino-5-(tri)
• methanol 3mL
• hydrochloric acid/methanol 6mL, 4N
• Step 4 To 3-(2-(((1S,3S)-3-((3-aminopropyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl) 2-(7-aza Benzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (80mg, 0.21mmol) and N,N-diisopropylethylamine (300mg, 2.32mmol), the The reaction solution was stirred at 80°C for 16 hours.
• Step 2 Add iodomethane (6.97g, 49.1 mmol) and potassium carbonate (11.3g, 81.9mmol). The reaction solution was stirred at 25° C. for 16 hours under nitrogen protection. After the completion of the reaction as detected by TLC, the reaction solution was added with water (200 mL), and then extracted with ethyl acetate (150 mL ⁇ 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated.
• Step 3 In a dry 50mL three-necked flask, add aluminum trichloride (1.27g, 9.54mmol), and add 7-chloro-1H-indole-6-carboxylic acid methyl ester (1.00g, 4.77mmol) under nitrogen protection 1.2-dichloroethane (10mL) solution, this system was stirred at 80°C for 30 minutes, then 2,4-dichloro-5-(trifluoromethyl)pyrimidine (1.09g, 5.01mmol) was added in 1.2-bis Chloroethane (1 mL) solution, and the reaction solution was reacted at 80° C. for 16 hours.
• Step 4 In a 10mL microwave tube, add 7-chloro-3-(2-chloro-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (420mg , 1.08mmol), N-methylpyrrolidone (2mL), N,N-diisopropylethylamine (0.53mL, 3.23mmol) and tert-butyl ((1S,3S)-3-aminocyclopentyl)amino Formic acid ester (237mg, 1.18mmol), the reaction solution was dissolved by stirring at 80°C, and then microwaved at 130°C for 2 hours.
• Step 5 At room temperature, methyl 3-(2-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidine- 4-yl)-7-chloro-1H-indole-6-carboxylate (430mg, 0.72mmol) was dissolved in a mixed solution of hydrogen chloride/dioxane (4mL, 4M) and methanol (2mL), reacted at 15°C 16 hours.
• Step 6 3-(2-(((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-chloro-1H-indole -6-Carboxylic acid methyl ester (320mg, 0.71mmol) was dissolved in N,N-dimethylformamide (2mL), tert-butyl (4-bromobutyl) carbamate (178mg, 0.71mmol) and N,N-Diisopropylethylamine (0.58 mL, 3.53 mmol) was added sequentially and the reaction was stirred at 80°C for 16 hours.
• Step 7 Methyl 3-(2-((1S,3S)-3-((4-((tert-butoxycarbonyl)amino)butyl)amino)cyclopentyl)amino)-5-(trifluoro Methyl)pyrimidin-4-yl)-7-chloro-1H-indole-6-carboxylate (230mg, 0.37mmol) was dissolved in a mixed system of water (1mL), methanol (1mL) and tetrahydrofuran (1mL) , then lithium hydroxide monohydrate (77.2mg, 1.84mmol) was added and the reaction was stirred at 40°C for 16 hours.
• Step 8 3-(2-(((1S,3S)-3-((4-((tert-butoxycarbonylamino)pentylamino)cyclopentyl)amino)-5-(trifluoromethyl) Pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid (lithium salt, 340mg) was dissolved in hydrogen chloride/methanol (5mL, 4M), and the system was stirred at 25°C for 4 Hour.
• Step 9 To 3-(2-(((1S,3S)-3-((4-aminobutyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl) 2-(7-azabenzene Triazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (108mg, 0.28mmol) and N,N-diisopropylethylamine (0.24mL, 1.42mmol), the The reaction solution was stirred at 60°C for 16 hours.
• Step 1 3-(2-((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethyl Phosphoryl)-1H-indole-6-carboxylic acid methyl ester (750mg, 1.52mmol) in methanol solution (10mL) was successively added tert-butyl ((4-oxocyclohexyl)methyl)carbamate (414mg , 1.82mmol) and glacial acetic acid (0.1mL), the reaction solution was stirred and reacted at 25°C for 16 hours under the protection of nitrogen.
• the obtained crude product was prepared and purified by reverse phase (chromatographic column: Ultimate XB-C18, 50*250mm, 10um (PARP-02); mobile phase: acetonitrile-water (0.1% trifluoroacetic acid); flow rate: 70mL/min; column Temperature: 25°C) to obtain white solid methyl 3-(2-((1S,3S)-3-((tert-butoxycarbonyl)amino)methylcyclohexylamino)cyclopentylamino)-5-(trifluoro Methyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (P1: 340 mg, yield 32%).
• Step 2 To methyl 3-(2-(((1S,3S)-3-((4-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)amino)cyclopentyl)amino)- 5-(Trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (P2: 340 mg, 0.48 mmol) in THF (3 mL) and methanol A solution of lithium hydroxide monohydrate (101 mg, 2.40 mmol) in water (3 mL) was added to the mixed solution of lithium hydroxide (3 mL), and the reaction solution was stirred and reacted at 40° C. overnight.
• Step 3 3-(2-(((1S,3S)-3-((4-(((tert-butoxycarbonyl)amino)methyl)cyclohexyl)amino)cyclopentyl)amino at 25°C )-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid (300mg, 0.43mmol) in methanol (3mL) was added Hydrochloric acid/dioxane (3 mL, 4N) solution, the reaction solution was stirred at 25°C for 16 hours.
• Step 4 To 3-(2-(((1S,3S)-3-((4-(aminomethyl)cyclohexyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidine- 4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid (30mg, 0.05mmol) in N,N-dimethylformamide (5mL) solution was added 2-( 7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (19mg, 0.05mmol) and N,N-diisopropylethylamine (32mg, 0.25 mmol), the reaction solution was stirred at 70°C for 14 hours.
• Step 1 Under nitrogen protection, add tert-butyl ((1S,3S)-3-hydroxycyclobutyl)(methyl)aminomethyl ester (200mg, 1.0mmol) in dichloromethane (4mL) at 25°C Dess-Martin reagent (843mg, 2.0mmol) was added, and the reaction solution was stirred and reacted at 25°C for 3 hours under the protection of nitrogen. After the reaction was complete as detected by LCMS, the reaction solution was extracted with water (10 mL) and ethyl acetate (20 mL ⁇ 3), and the combined organic layer was washed with saturated brine (10 mL ⁇ 2), dried over anhydrous sodium sulfate, filtered and concentrated to dryness.
• tert-butyl ((1S,3S)-3-hydroxycyclobutyl)(methyl)aminomethyl ester 200mg, 1.0mmol
• Dess-Martin reagent 843mg, 2.0mmol
• Step 2 Under nitrogen protection, 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl] Amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (423mg, 0.85mmol) and ⁇ [(3-oxylidenecyclobutyl)methyl] Acetic acid (20 mg, 0.33 mmol) was added to a solution of 2-methylpropan-2-yl amino ⁇ methanoate (170 mg, 0.85 mmol) in methanol (3 mL), and the reaction was stirred at 25°C for 16 hours under nitrogen protection.
• Step 3 Under nitrogen protection, at 25 ° C, to ⁇ [(3- ⁇ [(1S,3S)-3-[(4- ⁇ 7-[dimethyl(oxygenide)- ⁇ 5-methylphosphoryl]- 6-(methoxycarbonyl)-1H-indol-3-yl ⁇ -5-(trifluoromethyl)pyrimidin-2-yl)amino]cyclopentyl]amino ⁇ cyclobutyl)methyl]amino ⁇
• lithium hydroxide monohydrate 117mg, 2.8mmol
• Step 4 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3- ⁇ [3-(2,2-dimethyl -4-Oxylidene-5-aza-3-oxahex-6-yl)cyclobutyl]amino ⁇ cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)- To 1H-indole-6-carboxylic acid (300mg, 0.45mmol) in methanol (5mL) was added 4M methanolic hydrogen chloride solution (10mL), and the reaction solution was heated to 40°C under nitrogen protection and stirred for 2 hours.
• Step five to 7-[dimethyl (oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3- ⁇ [3-(aminomethyl)cyclobutyl ]amino ⁇ cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (50mg, 0.089mmol) in N,N-dimethylformamide ( 5mL) solution was added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (34mg, 0.089mmol) and N,N-di Isopropylethylamine (0.059mL, 0.35mmol), the reaction solution was stirred at 60°C for 2h.
• Step 1 To a solution of ⁇ [(3S)-tetrahydro-1H-pyrrol-3-yl]amino ⁇ methanoic acid benzyl ester (225mg, 1.02mmol) in N,N-dimethylmethaneamide (6mL) successively [(5-Bromopentyl)amino]methanoate-2-methylpropan-2-yl ester (299mg, 1.12mmol) and N,N-diisopropylethylenediamine (528mg, 4.08mmol) were added, and the The reaction solution was stirred at 80° C. for 16 hours under a nitrogen atmosphere.
• Step 2 To ⁇ [(3S)-1-(2,2-dimethyl-4-oxoylidene-5-aza-3-oxadec-10-yl)tetrahydro-1H-pyrrole-3 -Byl]amino ⁇ methanoic acid benzyl ester (500mg, 1.23mmol) in methanol (8mL) solution was added palladium carbon (50mg, 10% content, 60% water content), the reaction solution was stirred at 25°C under a hydrogen atmosphere for reaction 2 Hour. LCMS detected that the reaction was complete.
• Step 3 To ( ⁇ 5-[(3S)-3-aminotetrahydro-1H-pyrrol-1-yl]pentyl ⁇ amino)methanoic acid-2-methylpropan-2-yl ester (334mg, 1.23mmol ) in N,N-dimethylmethaneamide (8mL) was sequentially added 3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-7-[dimethyl(oxygenide) - ⁇ 5-methylphosphoryl]-1H-indole-6-carboxylic acid methyl ester (507mg, 1.23mmol) and N,N-diisopropylethylenediamine (477mg, 3.69mmol), the reaction solution was under nitrogen protection The temperature was raised to 80° C.
• reaction solution was extracted with water (20 mL) and dichloromethane (30 mL ⁇ 5), and the combined organic layer was washed with saturated brine (20 mL ⁇ 2), dried over anhydrous sodium sulfate, filtered and concentrated to dryness.
• Step 4 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(3S)-1-(2,2-dimethyl-4-oxygenidene -5-Aza-3-oxadec-10-yl)tetrahydro-1H-pyrrol-3-yl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole- Lithium hydroxide monohydrate (173 mg, 4.13 mmol) was added to a mixed solution of methyl 6-formate (550 mg, 0.83 mmol) in methanol (3 mL), water (3 mL), and tetrahydrofuran (3 mL), and the reaction solution was heated under nitrogen protection The reaction was stirred at 40°C for 2 hours.
• Step 5 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(3S)-1-(2,2-dimethyl-4-oxygenidene -5-Aza-3-oxadec-10-yl)tetrahydro-1H-pyrrol-3-yl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole- 4M 1,4-dioxane hydrogen chloride solution (8 mL) was added to 6-formic acid (480 mg, 0.70 mmol), and the reaction solution was heated to 40° C. under nitrogen protection and stirred for 2 hours.
• 6-formic acid 480 mg, 0.70 mmol
• Step 6 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(3S)-1-(5-aminopentyl)tetrahydro-1H-pyrrole- 3-yl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (100 mg, 0.18 mmol) in N,N-dimethylformamide (5 mL) Add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (69mg, 0.18mmol) and N,N-diisopropyl Ethylamine (94mg, 0.12mmol), the reaction solution was stirred at 60°C for 16 hours.
• Step 1 Add ⁇ [(3S)-hexahydropyridin-3-yl]amino ⁇ methanoic acid benzyl ester (242mg, 1.03mmol) to a solution of N,N-dimethylformamide (8mL) at 25°C [(5-Bromopentyl)amino]methanoate-2-methylpropan-2-yl ester (250mg, 0.939mmol), N,N-diisopropylethylamine (363mg, 2.82mmol) were added to the reaction The solution was stirred and reacted at 80° C. for 6 hours under the protection of nitrogen.
• Step 2 Add ( ⁇ 5-[(3S)-3- ⁇ [(benzyloxy)carbonyl]amino ⁇ hexahydropyridin-1-yl]pentyl ⁇ amino)methanoic acid-2-methylpropan-2
• the -yl ester 250 mg, 0.596 mmol
• methanol 10 mL
• palladium on carbon 100 mg, 10% content, 60% aqueous
• Step 3 Add ( ⁇ 5-[(3S)-3-aminohexahydropyridin-1-yl]pentyl ⁇ amino)methanoic acid-2-methylpropan-2-yl ester (180mg, 0.63mmol ) in N,N-dimethylformamide (6mL) solution, add 3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-7-[dimethyl(oxyethylene )- ⁇ 5 -methylphosphoryl]-1H-indole-6-carboxylic acid methyl ester (272mg, 0.63mmol), N,N-diisopropylethylamine (244mg, 1.89 mmol), the reaction solution was protected under nitrogen The reaction was stirred at 80°C for 16 hours.
• Step 4 Adding 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(3S)-1-(2,2-dimethyl-4 -Oxylidene-5-aza-3-oxadec-10-yl)hexahydropyridin-3-yl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole
• Lithium hydroxide hydrate (185mg, 4.41mmol) was added to methanol (4mL), water (4mL) and tetrahydrofuran (4mL) of methyl-6-carboxylate (300mg, 0.441mmol), and the reaction solution was heated under nitrogen protection at 40 The reaction was stirred at °C for 16 hours.
• Step 5 Add 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(3S)-1-(2,2-dimethyl-4- Oxylidene-5-aza-3-oxadec-10-yl)hexahydropyridin-3-yl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-
• dioxane hydrochloride 1.95 mL, 4 mol/L
• Step six at room temperature to 7-[dimethyl (oxygenide)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(3S)-1-(5-aminopentyl)hexahydropyridine- 3-yl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (70mg, 0.12mmol) in N,N-dimethylformamide (5mL) Add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (56.3mg, 0.14mmol), N,N-diisopropyl Ethylamine (77.4 mg, 0.6 mmol), and the reaction liquid was reacted at 65° C.
• Step 1 7-[Dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl]amino ⁇ -5-( Trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (300mg, 0.61mmol) was dissolved in N,N-dimethylformamide (3mL), [(4-bromobutyl Base)amino]methanoic acid-2-methylpropan-2-yl ester (153mg, 0.61mmol) and N,N-diisopropylethylamine (0.30mL, 1.83mmol) were added sequentially, and the reaction was stirred at 80°C for 16 Hour.
• Step 2 Add 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(2,2-dimethyl-4 -Oxylidene-5-aza-3-oxanon-9-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6 - Methyl formate (150 mg, 0.225 mmol) was dissolved in methanol (3 mL), then paraformaldehyde (45.0 mg, 1.13 mmol), palladium on carbon (10 mg, 10% content, 60% water) and glacial acetic acid (0.1 mL) were added , and stirred the reaction at 70°C for 5 hours.
• Step 4 Add 1,4-dioxane hydrochloride solution (5 mL, 4M) to the reaction solution in the previous step, and stir the system at 10° C. for 4 hours.
• the solvent was directly spin-dried, and the obtained crude product was prepared by reverse phase (chromatographic column: Ultimate XB-C18, 50*250mm, 10um (PARP-02); mobile phase: acetonitrile-water (0.1% bicarbonate ammonium); flow rate: 70mL/min; column temperature: 25°C) to obtain white solid 7-[dimethyl(oxyethylene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S) -3-[(4-aminobutyl)(methyl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (20mg, Yield: 13%).
• LCMS m/
• Step 5 To 7-[dimethyl (oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(4-aminobutyl)(methyl )amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (20mg, 0.04mmol) in N,N-dimethylformamide ( 3mL) solution was added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (14.8mg, 0.04mmol) and N,N- Diisopropylethylamine (0.03mL, 0.18mmol), the reaction solution was stirred at 60°C for 16 hours.
• Step 1 In a 250mL three-necked flask, sequentially add N-[(benzyloxy)carbonyl]glycine (20g, 95.6mmol), tetrahydrofuran (130mL), magnesium chloride (4.55g, 47.8mmol), carbonyldiimidazole (15.5 g, 95.6mmol), [(3-methoxy-1,3-dioxylidene)oxy] potassium (12.2g, 71.7mmol), the reaction solution was replaced with nitrogen, refluxed at 70°C for 4 hours, TLC The detection response is complete.
• Step 2 Dissolve benzyl [(1-methoxy-1,3-dioxylidenebut-4-yl)amino]methanoate (1.50g, 5.66mmol) in dichloromethane (30mL), and Add diethylaminosulfur trifluoride (1.00g, 6.22mmol) dropwise at 0°C, stir at 15°C for 16 hours, then add diethylaminosulfur trifluoride (1.00g, 6.22mmol) at 15°C, It was then stirred at 15°C for 4 hours.
• Step 3 Dissolve benzyl (3,3-difluoro-1-methoxy-1-oxyylidenebut-4-yl)amino]methanoate (130mg, 0.45mmol) in tetrahydrofuran (3mL), and A solution of lithium aluminum hydride in tetrahydrofuran (1 mL, 1 M) was added dropwise at 0°C, and the reaction was stirred at 10°C for 2 hours.
• Step 4 Dissolve [(2,2-difluoro-4-hydroxybutyl)amino]methanoic acid benzyl ester (120mg, 0.46mmol) in dichloromethane (1mL), then add pyridine (1mL) and p Methanesulfonyl chloride (265mg, 1.39mmol), the system was stirred at 10°C for 3 hours.
• Step 5 In a 10mL single-necked bottle, 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl ]Amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (173mg, 0.35mmol) was dissolved in N,N-dimethylformamide (3mL) , then added 4-methylbenzenesulfonic acid-6,6-difluoro-3-oxyylidene-1-phenyl-4-aza-2-oxahoct-8-yl ester (120mg, 0.29mmol) and N,N-Diisopropylethylamine (0.24 mL, 1.45 mmol).
• Step 6 Add 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(6,6-difluoro-3- Oxylidene-1-phenyl-4-aza-2-oxahoct-8-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H- Methyl indole-6-carboxylate (90mg, 0.12mmol) was dissolved in a mixed system of water (1mL), methanol (1mL) and tetrahydrofuran (1mL), and then lithium hydroxide monohydrate (5.13mg, 0.12mmol) was added, The reaction was stirred at 40°C for 16 hours.
• Step 7 In a 10mL single-necked bottle, add 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(6, 6-Difluoro-3-oxyylidene-1-phenyl-4-aza-2-oxahoct-8-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidine- 4-yl)-1H-indole-6-carboxylic acid (18 mg, 0.03 mmol) and trifluoroacetic acid (3 mL). Stir at 60°C for 1.5 hours.
• Step 8 To 2,2-difluoro-4- ⁇ [(1S,3S)-3-[(4- ⁇ 5-[(Z)-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphonium Base] methylidene] -4-[(1Z)-ethylidene]-1H-pyrrol-3-yl ⁇ -5-(trifluoromethyl)pyrimidin-2-yl)amino]cyclopentyl]amino ⁇
• 2-(7-azabenzotriazole)-N,N,N', N'-Tetramethyluronium hexafluorophosphate (9.95 mg, 0.03 mmol) and N,N-diisopropylethylamine (0.02 mL, 0.12 mmol).
• Step 1 [(1-Hydroxybut-4-yl)(methyl)amino]methanoic acid-2-methylpropan-2-yl ester (400mg, 1.97mmol) in tetrahydrofuran (8mL) at 25°C Carbon tetrabromide (981 mg, 3.74 mmol) and triphenylphosphine (1.24 g, 3.74 mmol) were added to the solution. The reaction solution was stirred and reacted at 25° C. for 16 hours under the protection of nitrogen.
• Step 2 Add [(1-bromobutyl-4-yl)(methyl)amino]methanoic acid-2-methylpropan-2-yl ester (240mg, 0.90mmol), 3-(2- ⁇ [(1S ,3S)-3-Aminocyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-7-[methyl(oxygenide)- ⁇ 5-methylphosphoryl]-1H-ind Indole-6-carboxylic acid methyl ester (447 mg, 0.90 mmol) was dissolved in acetonitrile (10 mL), and N,N-diisopropylethylamine (349 mg, 2.71 mmol) was added.
• Step 3 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(2,2,5- Trimethyl-4-oxyethylene-5-aza-3-oxanon-9-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H - To a solution of methyl indole-6-carboxylate (220mg, 0.32mmol) in methanol (8mL) was added successively paraformaldehyde (145mg, 1.62mmol), acetic acid (244mg, 1.89mmol), palladium on carbon (20mg, 10% content , 60% water).
• reaction was stirred at 80°C for 16 hours under hydrogen atmosphere. After the completion of the reaction detected by LCMS, an aqueous solution (50 mL) was added to the reaction solution, and extracted with dichloromethane (50 mL ⁇ 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated to dryness.
• Step 4 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-(2,2,5-tri Methyl-4-oxyylidene-5,10-diaza-3-oxaundecane-10-yl)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl) - To methyl 1H-indole-6-carboxylate (200 mg, 0.29 mmol) in methanol (3 mL), water (3 mL), tetrahydrofuran (3 mL) was added lithium hydroxide hydrate (121 mg, 2.88 mmol). The reaction was stirred at 40° C.
• Step 5 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-(2,2,5-tri Methyl-4-oxyylidene-5,10-diaza-3-oxaundecane-10-yl)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl) -
• To 1H-indole-6-carboxylic acid (180 mg, 0.26 mmol) in dioxane (4 mL) was added dioxane hydrochloride (1.32 mL, 5.29 mmol, 4 mol/L). The reaction was stirred at 25° C. for 16 hours under the protection of nitrogen.
• Step 6 To 7-[dimethyl (oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-(2,7-diazo N,N-dimethyl Add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (55mg, 0.15mmol), N, N-Diisopropylethylamine (78 mg, 0.61 mmol). The reaction was carried out at 60° C. for 18 hours under nitrogen protection.
• Step 1 Add 2-oxylidenetetrahydropyrrole-1-carboxylic acid-2-methylpropan-2-yl ester (2.00g, 10.8mmol) in tetrahydrofuran (30mL) at 0°C under nitrogen atmosphere Methylmagnesium bromide (15.1 mL, 15.1 mmol) was added dropwise. The reaction was stirred at 0 °C for 4 hours. After spotting the reaction to detect the completion of the reaction, water (30 mL) was added to quench the reaction, followed by extraction with ethyl acetate (100 mL ⁇ 3), the combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness.
• Step 2 Dissolve [(4-oxyethylenepentyl)amino]methanoic acid-2-methylpropan-2-yl ester (122mg, 0.61mmol) in tetrahydrofuran (5mL), then add isopropyl titanate ( 574mg, 2.0mmol), 7-[Dimethyl(oxysubunit)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl]amino ⁇ -5- (Trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (250mg, 0.51mmol), the reaction was stirred at 70°C for 16 hours, cooled to room temperature, to which was added cyanoborohydrogenation Sodium (159mg, 2.53mmol), stirred at 25°C for 6 hours.
• isopropyl titanate 574mg, 2.0mmol
• Step 4 Add hydrogen chloride/1,4-dioxane solution (5 mL, 4M) to the reaction solution in Step 3, and stir the system at 40° C. for 4 hours.
• the solvent was directly spin-dried to obtain the crude product, which was prepared by reverse phase (chromatographic column: Ultimate XB-C18, 50*250mm, 10um (PARP-02), mobile phase: acetonitrile-water (0.1% three Fluoroacetic acid), flow rate: 70mL/min, column temperature: 25°C) to obtain 7-[dimethyl(oxygenide)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3 -[(5-aminopent-2-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (150mg, yield 94 %), in the form of a white
• Step 5 7-[Dimethyl(oxyethylene)- ⁇ 5-methylphosphonyl]-3-(2- ⁇ [(1S,3S)-3-[(5-aminopent-2-yl)amino] Cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (70 mg, 0.12 mmol) in N,N-dimethylformamide (3 mL) Add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (47mg, 0.12mmol) and N,N-diisopropyl Ethylamine (64mg, 0.50mmol), the reaction solution was stirred at 80°C for 18h.
• Step 1 At 25°C, to 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl]amino ⁇
• [( 6-bromohexyl)amino]methanoic acid-2-methylpropan-2-yl ester (353 mg, 0.71 mmol)
• N,N-diisopropylethylamine 460 mg, 3.57 mmol
• Step 2 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(2,2-dimethyl Base-4-oxyethylene-5-aza-3-oxa-undecane-11-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H -To methyl indole-6-carboxylate (180 mg, 0.26 mmol) in methanol (3 mL), water (3 mL), tetrahydrofuran (3 mL) was added lithium hydroxide hydrate (109 mg, 2.59 mmol). The reaction was stirred at 40° C.
• Step 3 To 7-[dimethyl(oxygenide)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(2,2-dimethyl Base-4-oxyethylene-5-aza-3-oxa-undecane-11-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H - To methanol (4 mL) of indole-6-carboxylic acid (160 mg, 0.24 mmol) was added methanolic hydrochloride (1.12 mL, 4 mol/L). The reaction solution was stirred and reacted at 40° C. for 16 hours under the protection of nitrogen.
• Step 4 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(6-aminohexyl)amino) at room temperature ]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (70mg, 0.12mmol) in N,N-dimethylformamide (5mL) Add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (55.2mg, 0.15mmol), N,N-diisopropyl Ethylamine (78 mg, 0.61 mmol).
• Step 1 At room temperature, to (4 1 S,4 3 S)-1 7 -(dimethylphosphoryl)-2 5 -(trifluoromethyl)-1 1 H-8-oxa-3 ,5,11-Triaza-1(3,6)-indole-2(4,2)-pyrimidine-4(1,3)-cyclopentanecyclododecane-12-one 2,2, Paraformaldehyde (20 mg, 0.22 mmol) and isopropyl titanate (10.3 mg, 0.036 mmol) were successively added to a tetrahydrofuran solution (5 mL) of 2-trifluoroacetate (20 mg, 0.030 mmol). The reaction was stirred at 60° C. for 14 hours under the protection of nitrogen.
• Step 1 Add [(6-bromohexyl)amino]methanoic acid-2-methylpropan-2-yl ester (300mg, 1.28mmol) to N,N-dimethylformamide (8mL) at 25°C Benzyl ⁇ [(3S)-hexahydropyridin-3-yl]amino ⁇ methanoate (395 mg, 1.41 mmol), N,N-diisopropylethylamine (662 mg, 5.12 mmol) were added to the solution. The reaction solution was stirred and reacted at 80° C. for 6 hours under the protection of nitrogen.
• Step 2 Add ⁇ [(3S)-1-(2,2-dimethyl-4-oxyylidene-5-aza-3-oxaundecane-11-yl)hexahydropyridine-3- Benzyl]amino ⁇ methanoate (555 mg, 1.17 mmol) was dissolved in methanol (10 mL) and palladium on carbon (60 mg, 10% content, 60% aqueous) was added. The reaction solution was stirred and reacted at 25° C. for 3 hours under the protection of hydrogen.
• Step 3 Add ⁇ 6-[(3S)-3-aminohexahydropyridin-1-yl]hexyl ⁇ amino (280mg, 0.94mmol) to a solution of N,N-dimethylformamide (8mL) at room temperature Add 3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-7-[dimethyl(oxygenide)- ⁇ 5-methylphosphoryl]-1H-indole-6 in sequence -Methyl formate (404mg, 0.94mmol), N,N-diisopropylethylamine (483mg, 3.74mmol), the reaction solution was stirred and reacted at 80°C for 16 hours under the protection of nitrogen.
• Step 4 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(3S)-1-(2,2-dimethyl-4 -Oxylidene-5-aza-3-oxaundecane-11-yl)hexahydropyridin-3-yl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H- Add lithium hydroxide hydrate (74mg, 1.75mmol) to methyl indole-6-carboxylate (243mg, 0.35mmol) in methanol (3mL), water (3mL) and tetrahydrofuran (3mL), and the reaction solution is under nitrogen protection The reaction was stirred at 40°C for 14 hours.
• Step five to 7-[dimethyl (oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(3S)-1-(2,2-dimethyl-4 -Oxylidene-5-aza-3-oxaundecane-11-yl)hexahydropyridin-3-yl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-
• methanol 4methanol
• hydrogen chloride/methanol solution 6mL, 4mol/L
• Step 6 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(3S)-1-(6-aminohexyl)hexahydropyridine- 3-yl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (100mg, 0.17mmol) in N,N-dimethylformamide (3mL) Add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (65.4mg, 0.17mmol), N,N-diisopropyl Ethylamine (88.9 mg, 0.69 mmol), and the reaction liquid was reacted at 80° C.
• Step 1 At 25°C, to ⁇ [2-(tetrahydro-1H-pyrrol-3-yl)ethyl]amino ⁇ methane acid-2-methylpropan-2-yl ester (200mg, 0.93mmol) N,N-Dimethylformamide (6mL) solution was added [(2-bromoethyl)amino]benzyl methane (241mg, 0.93mmol), N,N-diisopropylethylamine (602mg, 4.67 mmol). The reaction solution was stirred and reacted at 80° C. for 5 hours under the protection of nitrogen.
• Step 2 To ( ⁇ 2-[1-(3-Oxylidene-1-phenyl-4-aza-2-oxahex-6-yl)tetrahydro-1H-pyrrole-3 -2-methylprop-2-yl-]ethyl ⁇ amino)methanoate (200 mg, 0.51 mmol) in methanol was added Pd/C (20 mg, 10% content, 60% aqueous). The reaction solution was stirred and reacted at 25° C. for 5 hours under the protection of hydrogen.
• Step 3 To ( ⁇ 2-[1-(2-aminoethyl)tetrahydro-1H-pyrrol-3-yl]ethyl ⁇ amino)methanoic acid-2-methylpropan-2-yl at room temperature Ester (100mg, 0.39mmol) in N,N-dimethylformamide (6mL) was added 3-[2-chloro-5-(trifluoromethyl)pyrimidin-4-yl]-7-[dimethyl (Oxylidene)- ⁇ 5-methylphosphonyl]-1H-indole-6-carboxylic acid methyl ester (168 mg, 0.39 mmol), N,N-diisopropylethylamine (150 mg, 1.17 mmol).
• Step 4 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-[2-( ⁇ 2-[3-(2,2-dimethyl-4- Oxylidene-5-aza-3-oxahept-7-yl)tetrahydro-1H-pyrrol-1-yl]ethyl ⁇ amino)-5-(trifluoromethyl)pyrimidin-4-yl] - To methyl 1H-indole-6-carboxylate (220 mg, 0.34 mmol) in methanol (3 mL), water (3 mL), tetrahydrofuran (3 mL) was added lithium hydroxide hydrate (141 mg, 3.37 mmol). The reaction was stirred at 40° C.
• Step five at room temperature, to 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-[2-( ⁇ 2-[3-(2,2-dimethyl-4 -Oxylidene-5-aza-3-oxahept-7-yl)tetrahydro-1H-pyrrol-1-yl]ethyl ⁇ amino)-5-(trifluoromethyl)pyrimidin-4-yl ]-1H-indole-6-carboxylic acid (200mg, 0.31mmol) in 1,4-dioxane (4mL) was added hydrochloride 1,4-dioxane (0.78mL, 3.13mmol, 4mol/L) .
• Step 6 3-[2-( ⁇ 2-[3-(2-aminoethyl)tetrahydro-1H-pyrrol-1-yl]ethyl ⁇ amino)-5-(trifluoromethyl) at room temperature Base) pyrimidin-4-yl]-7-[dimethyl(oxyethylene)- ⁇ 5-methylphosphoryl]-1H-indole-6-carboxylic acid (70mg, 0.11mmol) N,N-dimethyl Add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (51mg, 0.13mmol), N,N - Diisopropylethylamine (43 mg, 0.33 mmol).
• Step 1 In a 50mL single-necked bottle, to (1-hydroxy-6-aza-3-oxahept-6-yl)methanoic acid-2-methylpropan-2-yl ester (300mg, 1.37mmol) Imidazole (140mg, 2.05mmol), triphenylphosphine (431mg, 1.64mmol) and iodine (521mg, 2.05mmol) were sequentially added to the dichloromethane (10mL) solution. Stir at 5°C for 2 hours under nitrogen protection.
• Step 2 In a 50mL single-necked bottle, to (1-iodo-6-aza-3-oxahept-6-yl)methanoic acid-2-methylpropan-2-yl ester (350mg, 1.06mmol) Add 7-[dimethyl(oxygenide)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3- Aminocyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (553mg, 1.12mmol) and N,N-diisopropylethylamine (0.88 mL, 5.32 mmol).
• Step 3 Add 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(2,2,5-trimethyl -4-Oxylidene-5-aza-3,8-dioxadec-10-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H - Methyl indole-6-carboxylate (610mg, 0.88mmol) was dissolved in a mixed system of water (3mL), methanol (3mL) and tetrahydrofuran (3mL).
• Step 4 Add methanol solution of hydrochloric acid (5 mL, 4M) to the reaction solution in the previous step, and stir at 5° C. for 5 hours. After the LCMS detection is complete, the reaction solution is concentrated, and the crude product is prepared by reverse phase (chromatographic column: Ultimate XB-C18, 50*250mm, 10um (PARP-02), mobile phase: acetonitrile-water (0.1% trifluoroacetic acid), flow rate: 70mL/min, column temperature: 25°C) to obtain white solid 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-(6 -Aza-3-oxepept-1-ylamino)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (260mg, 2 steps Yield 51%).
• LCMS m/z (ES
• Step 5 To 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-(6-aza-3-oxapeptyl -1-ylamino)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (260mg, 0.45mmol) of N,N-dimethyl Add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (187mg, 0.49mmol) and N to the formamide (10mL) solution successively, N-Diisopropylethylamine (0.37 mL, 2.23 mmol).
• Step 1 In a 10mL single-necked bottle, add (4 1 S,4 3 S)-1 7 -(dimethylphosphoryl)-11-methyl-2 5 -(trifluoromethyl)-1 1 H- 8-oxa-3,5,11-triaza-1(3,6)-indole-2(4,2)-pyrimidine-4(1,3)-cyclopentanecyclododecane-12 Paraformaldehyde (9.30 mg, 0.31 mmol) and tetraisopropyl titanate (35.2 mg, 0.12 mmol) were successively added to a solution of -ketone trifluoroacetate (35 mg, 0.06 mmol) in tetrahydrofuran (3 mL).
• Step 1 At 25°C, to a solution of methyl 7-bromo-1H-indole-6-carboxylate (1.0g, 3.9mmol) in N,N dimethylformamide (10mL), add tetrakis(triphenyl Phosphine)palladium (0.45g, 0.39mmol) and zinc cyanide (0.46g, 3.9mmol). Blow nitrogen for one minute and put in microwave at 150°C for 1 hour.
• Step 2 Add N-iodobutane to a solution of methyl 7-cyano-1H-indole-6-carboxylate (800mg, 4.0mmol) in N,N dimethylformamide (10mL) at 25°C Imide (1.35 g, 6.0 mmol). Nitrogen was replaced, stirred at 40°C for 3 hours, and the reaction was complete as detected by LCMS. Water (30 mL) was added to the reaction solution, extracted with ethyl acetate (40 mL ⁇ 2), the combined organic layers were washed with saturated brine (40 mL ⁇ 4), dried over anhydrous sodium sulfate, filtered and concentrated.
• Step 3 At 25°C, add 7-cyano-3-iodo-1H-indole-6-carboxylic acid methyl ester (740mg, 2.27mmol) and 4,4,5,5-tetramethyl-1,3, To a solution of 2-dioxaborolane (871mg, 6.81mmol) in tetrahydrofuran (15mL), tris(dibenzylideneacetone) dipalladium (312mg, 0.34mmol), 2-dicyclohexylphosphine- 2',4',6'-Triisopropylbiphenyl (108mg, 0.227mmol) and triethylamine (689mg, 6.81mmol).
• Step 4 At 25°C, to 6-formyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborol-2-yl)-1H-ind Indole-7-carbonitrile methane (600 mg, 1.84 mmol) and ⁇ [(1S,3S)-3- ⁇ [4-chloro-5-(trifluoromethyl)pyrimidin-2-yl]amino ⁇ cyclopentyl] Amino ⁇ methanoic acid-2-methylpropan-2-yl ester (583mg, 1.53mmol) in tetrahydrofuran solution (10mL) was successively added chloro[(n-butylbis(1-adamantyl)phosphine)-2-( 2-Aminobiphenyl)] palladium (II) (102 mg, 0.153 mmol) and potassium phosphate (976 mg, 4.6 mmol) in water (2 mL).
• Step 5 At 25°C, to 7-cyano-3-(2- ⁇ [(1S,3S)-3-( ⁇ [(2-methylpropan-2-yl)oxy]carbonyl ⁇ amino) ring Pentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (500mg, 0.92mmol) in dichloromethane (5mL) was added trifluoroacetic acid ( 2.5mL). The reaction was carried out at 20° C. for 3 hours, and the reaction was detected by LCMS to be complete.
• Step 6 At room temperature, to 7-cyano-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)- 1H-Indole-6-carboxylic acid methyl ester (100mg, 0.225mmol) in N,N-dimethylformamide solution (2mL), add [(4-bromobutyl)amino]methane acid-2-methane 2-ylpropan-2-yl ester (56mg, 0.225mmol) and N,N-diisopropylethylamine (116mg, 0.9mmol).
• Step 7 To 7-cyano-3-(2- ⁇ [(1S,3S)-3-[(2,2-dimethyl-4-oxoylidene-5-aza-3-oxanonyl -9-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (70mg, 0.09mmol) in tetrahydrofuran (3mL) A solution of lithium hydroxide monohydrate (19.0 mg, 0.45 mmol) in water (3 mL) was added to the mixed solution of methanol (3 mL). The reaction was stirred at 40° C.
• Step 8 To 7-cyano-3-(2- ⁇ [(1S,3S)-3-[(2,2-dimethyl-4-oxyylidene-5-aza-3-oxanonyl -9-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (20mg, 0.033mmol) in dichloromethane (0.9mL ) solution was added trifluoroacetic acid (0.1 mL). The reaction was stirred at 15°C for 45 minutes. After the reaction was detected by LCMS, N,N-diisopropylethylamine was added to the reaction solution to adjust the pH to 8.
• Step 9 To 7-cyano-3-(2- ⁇ [(1S,3S)-3-[(4-aminobutyl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidine -4-yl)-1H-indole-6-carboxylic acid (12mg, 0.02mmol) in N,N-dimethylformamide (2mL) solution, add 2-(7-azabenzotriazole )-N,N,N',N'-tetramethyluronium hexafluorophosphate (15 mg, 0.04 mmol) and N,N-diisopropylethylamine (21 mg, 0.16 mmol).
• Step 1 Add [(4-hydroxybutyl)(methyl)amino]methane acid to a solution of triphenylphosphine (129mg, 0.49mmol) in dichloromethane (3mL) successively under nitrogen atmosphere at 0°C- 2-Methylpropan-2-yl ester (100mg, 0.49mmol), elemental iodine (125mg, 0.49mmol), imidazole (50mg, 0.74mmol). After stirring at 0°C for 5 minutes and warming to room temperature, the reaction was stirred at room temperature for 4 hours.
• triphenylphosphine 129mg, 0.49mmol
• dichloromethane 3mL
• elemental iodine 125mg, 0.49mmol
• imidazole 50mg, 0.74mmol
• Step 2 7-[Dimethyl(oxygenide)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl]amino ⁇ -5-(three Fluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (27 mg, 0.054 mmol) was dissolved in N,N-dimethylformamide (2 mL). Add [(4-iodobutyl)(methyl)amino]methanoic acid-2-methylprop-2-yl ester (20mg, 0.065mmol), N,N-diisopropylethylamine (28mg, 0.218 mmol).
• Step 3 Add 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(2,2,5-trimethyl- 4-Oxylidene-5-aza-3-oxanon-9-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-
• Methyl 6-carboxylate 110mg, 0.162mmol
• Lithium hydroxide (1 H2O, 34 mg, 0.808 mmol
• Step 4 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(2,2,5-trimethyl- 4-Oxylidene-5-aza-3-oxanon-9-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole- Hydrogen chloride/methanol solution (5 mL, 4M) was added to 6-formic acid (110 mg, 0.164 mmol), and the system was stirred at 40°C for 4 hours.
• Step five to 7-[dimethyl (oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3- ⁇ [4-(methylamino)butyl] Amino ⁇ cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (60mg, 0.106mmol) in N,N-dimethylformamide (5mL ) solution was added 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (40mg, 0.106mmol) and N,N-diiso Propylethylamine (68 mg, 0.530 mmol).
• Step 1 To a solution of methyl 4-((benzyloxy)carbonyl)amino)-3-oxobutanoate (5.0 g, 18.9 mmol) in THF (100 mL) at 5° C. was added lithium aluminum hydride in THF solution (28.3 mL, 1 M, 28.3 mmol). The reaction solution was stirred and reacted at 5° C. for 2 hours under the protection of nitrogen.
• Step 2 Add imidazole (0.68g, 10.04mmol) to a solution of benzyl (2,4-dihydroxybutyl) carbamate (1.2g, 5.02mmol) in dichloromethane (50mL) at 5°C , tert-butyldiphenylchlorosilane (1.38 g, 5.02 mmol). The reaction was stirred at 5° C. for 3 hours under the protection of nitrogen.
• Step 3 Benzyl (4-((tert-butyldiphenylsilyl)oxy)-2-hydroxybutyl)carbamate (2.2g, 4.61mmol) was dissolved in dichloromethane (30mL), Diethylaminosulfur trifluoride (1.1 g, 6.92 mmol) was then added slowly at 0°C and the reaction was stirred at 5°C for 3 hours.
• Step 4 Dissolve benzyl (4-((tert-butyldiphenylsilyl)oxy)-2-fluorobutyl)carbamate (700mg, 1.46mmol) in N,N-dimethylformaldehyde To the amide (5 mL), cesium fluoride (888 mg, 5.84 mmol) was added. The reaction was stirred at 5°C for 18 hours.
• Step 5 Dissolve benzyl (2-fluoro-4-hydroxybutyl) carbamate (300mg, 1.24mmol) in dichloromethane (10mL) in a 50mL single-necked bottle, add imidazole (126mg, 1.86mmol) , triphenylphosphine (390mg, 1.50mmol) and elemental iodine (381mg, 1.50mmol). The reaction was stirred at 10°C for 1 hour.
• Step 6 3-(2-((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(dimethylphosphoryl) -1H-indole-6-carboxylic acid methyl ester (220mg, 0.44mmol) was dissolved in N,N-dimethylformamide (5mL), and benzyl(2-fluoro-4-iodobutyl)aminomethyl ester (156mg, 0.44mmol) and N,N-diisopropylethylamine (286mg, 2.20mmol). Stir at 100°C for 18 hours.
• Step 7 Methyl 3-(2-((1S,3S)-3-((benzyloxy)carbonyl)amino)-3-fluorobutylamino)cyclopentylamino)-5-(trifluoromethyl yl)pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylate (290mg, 0.40mmol) was dissolved in methanol (5mL), tetrahydrofuran (5mL) and water (5mL ) into the mixed system, lithium hydroxide monohydrate (84mg, 2.00mmol) was added. The reaction was stirred at 40°C for 18 hours.
• Step 8 To 3-(2-((1S,3S)-3-((benzyloxy)carbonyl)amino)-3-fluorobutylamino)cyclopentylamino)-5-(trifluoromethyl) Add trifluoroacetic acid (5 mL) to a reaction flask of pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid (300 mg, 0.43 mmol), and stir at 60° C. for 3 h.
• Step 9 To 3-(2-((1S,3S)-3-((4-amino-3-fluorobutyl)amino)cyclopentyl)amino)-5-(trifluoromethyl) at room temperature )pyrimidin-4-yl)-7-(dimethylphosphoryl)-1H-indole-6-carboxylic acid (70mg, 0.12mmol) in N,N-dimethylformamide (5mL) was added successively 2 -(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (55mg, 0.15mmol), N,N-diisopropylethylamine (78mg ,0.61mmol).
• Step 10 Reverse phase the crude product obtained in the previous step (chromatographic column: Ultimate XB-C18, 50*250mm, 10um (PARP-02), mobile phase: acetonitrile-water (0.1% trifluoroacetic acid), flow rate: 70mL /min, column temperature: 25°C) to prepare and purify two isomeric compounds.
• One of the isomeric compounds is arbitrarily designated as (4 1 S,4 3 S,8R)-1 7 -(dimethylphosphoryl)-8-fluoro-2 5 -(trifluoromethyl)-1 1 H- 3,5,10-Triaza-1(3,6)-indole-2(4,2)-pyrimidine-4(1,3)-cyclopentanecycloundecane-11-one (4 1 S,4 3 S,8R)-1 7 -(dimethylphosphoryl)-8-fluoro-2 5 -(trifluoromethyl)-1 1 H-3,5,10-triaza-1(3,6)-indola-2( 4,2)-py rimidina-4(1,3)-cyclopentanacycloundecaphan-11-one (Z68-1, 4 mg, yield 6%, trifluoroacetate salt), white solid.
• Another isomeric compound is arbitrarily designated as (4 1 S,4 3 S,8S)-1 7 -(dimethylphosphoryl)-8-fluoro-2 5 -(trifluoromethyl)-1 1 H- 3,5,10-Triaza-1(3,6)-indole-2(4,2)-pyrimidine-4(1,3)-cyclopentanecycloundecane-11-one (4 1 S,4 3 S,8S)-1 7 -(dimethylphosphoryl)-8-fluoro-2 5 -(trifluoromethyl)-1 1 H-3,5,10-triaza-1(3,6)-indola-2( 4,2)-pyrimidina-4(1,3)-cyclopentanacycloundecaphan-11-one (Z68-2, 3 mg, yield 5%, trifluoroacetate salt).
• Step 1 In a 250mL three-necked flask, add N-[(benzyloxy)carbonyl]-N-methylglycine (10.8g, 48.4mmol), tetrahydrofuran (100mL) and carbonyldiimidazole (7.8g, 48.4mmol) in sequence .
• the reaction solution was replaced with nitrogen, stirred at 10°C for 12 hours, magnesium chloride (2.3g, 24.2mmol) and monomethyl malonate potassium salt (7.6g, 48.4mmol) were added, the temperature was raised to 60°C and stirred for 6 hours, TLC detected that the reaction was complete .
• Step 2 Dissolve benzyl (3,5-dioxylidene-7-aza-2-oxahoct-7-yl)methanoate (5.8g, 20.8mmol) in dichloromethane (50mL), Diethylaminosulfur trifluoride (5.0 g, 31.2 mmol) was added dropwise at 0°C. Stir at 25°C for 16 hours.
• Step 3 Dissolve benzyl (5,5-difluoro-3-oxylidene-7-aza-2-oxahoct-7-yl)methanoate (1.4g, 4.7mmol) in tetrahydrofuran (20mL ), added dropwise a tetrahydrofuran solution (4.7 mL, 1M) of lithium aluminum tetrahydrogen at 0°C, and stirred at 10°C for 2 hours.
• Step 4 To a solution of benzyl [(2,2-difluoro-4-hydroxybutyl)(methyl)amino]methanoate (400mg, 1.46mmol) in pyridine (5mL), add p-methylsulfonyl chloride (558mg, 2.93mmol), stirred at 10°C for 3 hours.
• Step 5 7-[Dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl]amino ⁇ -5-( Trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (426mg, 0.86mmol) was dissolved in N,N-dimethylformamide (7mL), and 4-methylbenzene Sulfonic acid-6,6-difluoro-4-methyl-3-oxylidene-1-phenyl-4-aza-2-oxahoct-8-yl ester (350mg, 0.82mmol) and N, N-Diisopropylethylamine (0.68 mL, 4.09 mmol).
• Step 6 Add 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(6,6-difluoro-4- Methyl-3-oxoylidene-1-phenyl-4-aza-2-oxa-oct-8-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidine-4- base)-1H-indole-6-carboxylic acid methyl ester (330mg, 0.46mmol) was dissolved in a mixed system of water (3mL), methanol (3mL) and tetrahydrofuran (3mL), and lithium hydroxide monohydrate (96.6mg, 2.30 mmol), stirred at 40°C for 16 hours.
• Step 7 To 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(6,6-difluoro-4- Methyl-3-oxoylidene-1-phenyl-4-aza-2-oxa-oct-8-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidine-4- (1)-1H-indole-6-carboxylic acid (350mg, 0.48mmol) was added trifluoroacetic acid (5mL).
• Step 8 To 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3- ⁇ [3,3-difluoro-4- N,N - Add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (149mg, 0.39mmol ) and N,N-diisopropylethylamine (0.30 mL, 1.78 mmol).
• Step 1 In a 10mL single-necked bottle, add (4 1 S,4 3 S)-1 7 -(dimethylphosphoryl)-8,8-difluoro-10-methyl-2 5 -(trifluoro Methyl)-1 1 H-3,5,10-Triaza-1(3,6)-indole-2(4,2)-pyrimidine-4(1,3)-cyclopentanecycloundec Paraformaldehyde (9 mg, 0.10 mmol) and tetraisopropyl titanate (28 mg, 0.10 mmol) were successively added to a solution of alkan-11-one (14 mg, 0.02 mmol, trifluoroacetate) in tetrahydrofuran (3 mL).
• Step 1 Add L-proline (0.18g , 1.6mmol), ketone iodide (0.95g, 0.8mmol) and sodium methylsulfite (1.1g, 9.5mmol). The reaction was stirred at 90° C. for 16 hours under the protection of nitrogen. After LCMS detected that the reaction was complete, slowly add saturated ammonium chloride aqueous solution (30mL) to the reaction solution to quench, and extract with ethyl acetate (30mL ⁇ 3), the combined organic layer was washed 3 times with saturated brine, and anhydrous sodium sulfate Dry, filter and concentrate.
• Step 2 Add N- Iodosuccinimide (1.5 g, 6.6 mmol). The reaction solution was stirred at room temperature for 16 hours under nitrogen protection. After the reaction was complete as detected by LCMS, the reaction solution was added with water (30 mL), and then extracted with ethyl acetate (30 mL ⁇ 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by filtration.
• Step 3 At room temperature, add 3-iodo-7-(methylsulfonyl)-1H-indole-6-carboxylic acid methyl ester (1.8g, 4.7mmol), tris(dibenzylidene Dipalladium (0.44g, 0.47mmol), 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (0.45g, 0.94mmol), pinacol borane (0.91g, 7.1 mmol), triethylamine (1.4g, 14.1mmol) and tetrahydrofuran (15mL) solution. Stir at 90°C for 16 hours under nitrogen protection.
• Step 4 At room temperature, sequentially add methyl 7-(methylsulfonyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxy Boropentan-2-yl)-1H-indole-6-carboxylate (1.1g, 2.9mmol), chloro[(n-butylbis(1-adamantyl)phosphine)-2-(2- Aminobiphenyl)] palladium (II) (0.19g, 0.29mmol), potassium phosphate (1.8g, 8.7mmol), tert-butyl ((1S,3S)-3-((4-chloro-5-(trifluoro A mixed solution of methyl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (1.1 g, 2.9 mmol), tetrahydrofuran (25 mL) and water (5 mL).
• Step 5 At room temperature, methyl 3-(2-((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5-(trifluoromethyl)pyrimidine-4 -yl)-7-(methylsulfonyl)-1H-indole-6-carboxylate (570mg, 0.95mmol) was dissolved in a mixed solution of hydrogen chloride/ethyl acetate (5mL, 4M) and ethyl acetate (3mL) , reacted at 30°C for 16 hours.
• a mixed solution of hydrogen chloride/ethyl acetate 5mL, 4M
• ethyl acetate 3mL
• Step 6 At room temperature, to 3-(2-((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(methylsulfonyl )-1H-indole-6-carboxylic acid methyl ester (300mg, 0.60mmol) in N,N-dimethylformamide (10mL) solution was added successively tert-butyl (2-(2-iodoethoxy) Ethyl)(methyl)carbamate (197mg, 0.60mmol) and N,N-diisopropylethylamine (310mg, 2.40mmol), stirred at 80°C for 16 hours.
• Step 7 Methyl 3-(2-((1S,3S)-3-((2-(2-((tert-butoxycarbonyl)(methyl)amino)ethoxy)ethyl)amino) ring Pentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(methylsulfonyl)-1H-indole-6-carboxylate (180mg, 0.26mmol) was dissolved in water ( 3 mL), methanol (3 mL) and tetrahydrofuran (3 mL), then added lithium hydroxide monohydrate (55 mg, 1.3 mmol), and stirred at 40°C for 16 hours.
• Step 8 Add 3-(2-((1S,3S)-3-((2-(2-((tert-butoxycarbonyl)(methyl)amino)ethoxy)ethyl)amino)cyclopentyl )amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-(methylsulfonyl)-1H-indole-6-carboxylic acid (176mg, 0.26mmol) was dissolved in methanol (5mL) , then added hydrochloric acid/methanol (5 mL, 4M), and stirred at 40° C. for 16 hours.
• Step 9 To 3-(2-((1S,3S)-3-((2-(methylamino)ethoxy)ethyl)amino)cyclopentylamino)-5-(trifluoromethyl)pyrimidine -4-yl)-7-(methylsulfonyl)-1H-indole-6-carboxylic acid (110mg, 0.16mmol, trifluoroacetate) in N,N-dimethylformamide (10mL) solution Add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (61mg, 0.16mmol) and N,N-diisopropyl ethyl Amine (83mg, 0.64mmol), stirred at 60°C for 16h.
• Step 10 In a 10mL single-necked bottle, add (4 1 S,4 3 S)-11-methyl-1 7 -(methylsulfonyl)-2 5 -(trifluoromethyl)-1 1 H- 8-oxa-3,5,11-triaza-1(3,6)-indole-2(4,2)-pyrimidine-4(1,3)-cyclopentanecyclododecane-12 Add paraformaldehyde (20mg, 0.221mmol) and tetraisopropyl titanate (63mg, 0.221mmol) successively to tetrahydrofuran (10mL) solution of -ketone (30mg, 0.044mmol, trifluoroacetate), and the reaction solution is Stir at 70°C for 14 hours, lower the reaction solution to 30°C, add sodium cyanoborohydride (41 mg, 0.660 mmol), and continue stirring at 30°C for 2 hours.
• paraformaldehyde (20mg, 0.221mmol) and
• Step 1 To a solution of N-[(benzyloxy)carbonyl]glycine (100.0 g, 478.49 mmol) in tetrahydrofuran (1000 mL) was added carbonyldiimidazole (85.3 g, 526.34 mmol) at 15°C. The reaction solution was stirred and reacted at 15° C. for 18 hours under the protection of nitrogen. Magnesium chloride (22.8 g, 239.25 mmol), monomethyl malonate potassium salt (82.2 g, 526.34 mmol) were then added to the reaction liquid. The reaction solution was stirred and reacted at 45° C. for 6 hours under the protection of nitrogen.
• Step 2 Dissolve benzyl [(1-methoxy-1,3-dioxylidenebut-4-yl)amino]methanoate (85.3g, 321.89mmol) in tetrahydrofuran (900mL) at 15°C Lithium borohydride (14.0 g, 643.78 mmol) was added in portions to the solution. The reaction solution was stirred and reacted at 15° C. for 18 hours under the protection of nitrogen.
• Step 3 At 15°C, add imidazole (22.2g, 326.37 mmol), and then slowly added tert-butyldiphenylchlorosilane (69.0 g, 251.05 mmol) dropwise. The reaction solution was stirred and reacted at 15° C. for 2 hours under the protection of nitrogen.
• Step 4 Benzyl [(7-hydroxy-2,2-dimethyl-3,3-diphenyl-4-oxa-3-silaoct-8-yl)amino]methanoate (88.0 g, 184.49mmol) was dissolved in dichloromethane (900mL), then diethylaminosulfur trifluoride (38.7g, 239.83mmol) was slowly added dropwise at room temperature. The reaction was stirred at 15°C for 2 hours.
• Step five benzyl [(7-fluoro-2,2-dimethyl-3,3-diphenyl-4-oxa-3-silaoct-8-yl)amino]methanoate (20.5 g, 42.80mmol) was dissolved in tetrahydrofuran (150mL), sodium hydrogen (5.1g, 60% content dispersed in mineral oil, 128.40mmol) was added in batches at room temperature, and then slowly added dropwise methyl iodide (12.2g, 85.60 mmol). The reaction was stirred at 15°C for 12 hours.
• Step 6 Benzyl (7-fluoro-2,2-dimethyl-3,3-diphenyl-9-aza-4-oxa-3-siladec-9-yl)methanoate (10.0g, 20.24mmol) was dissolved in a solution of N,N-dimethylformamide (60mL), then cesium fluoride (15.4g, 101.20mmol) was added, and stirred at 50°C for 8 hours.
• Step 7 Dissolve [(2-fluoro-4-hydroxybutyl)(methyl)amino]methanoic acid benzyl ester (4.4g, 17.25mmol) in a solution of dichloromethane (40mL) at room temperature , and then sequentially added imidazole (1.4g, 20.70mmol), triphenylphosphine (5.4g, 20.70mmol) and elemental iodine (5.3g, 20.70mmol). The reaction was stirred for an additional 2 hours at room temperature.
• Step 8 In a 250mL single-necked bottle, add 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl ]Amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid methyl ester (5.0g, 10.10mmol) was dissolved in N,N-dimethylformamide (70 mL ), then added benzyl [(2-fluoro-4-iodobutyl)(methyl)amino]methanoate (3.7g, 10.10mmol) and N,N-diisopropylethylamine (5.2g, 40.40 mmol), stirred at 100°C for 16 hours.
• Step 9 Add 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-[(6-fluoro-4-methyl- 3-Oxylidene-1-phenyl-4-aza-2-oxa-oct-8-yl)amino]cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)- 1H-Indole-6-methyl carboxylate (7.0g, 9.56mmol) was dissolved in a mixed system of water (20mL), methanol (20mL) and tetrahydrofuran (20mL), and then lithium hydroxide monohydrate (1.6g, 38.24 mmol), the reaction was stirred at 40°C for 16 hours.
• Step 11 To 7-[dimethyl(oxygenide)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3- ⁇ [3-fluoro-4-(methyl N,N- Add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2.6g, 6.85mmol ) and N,N-diisopropylethylamine (3.6g, 27.4mmol), the reaction solution was stirred at 60°C for 16h.
• One of the isomeric compounds is arbitrarily designated as (4 1 S,4 3 S,8R)-1 7 -(dimethylphosphoryl)-8-fluoro-10-methyl-2 5 -(trifluoromethyl) -1 1 H-3,5,10-Triaza-1(3,6)-indole-2(4,2)-pyrimidine-4(1,3)-cyclopentanecycloundecane-11 -Keto(4 1 S,4 3 S,8R)-1 7 -(dimethylphosphoryl)-8-fluoro-10-methyl-2 5 -(trifluoromethyl)-1 1 H-3,5,10-triaza-1( 3,6)-indola-2(4,2)-pyrimidina-4(1,3)-cyclopentanacycloundecaphan-11-one (Z76, 0.7g, yield 18%, trifluoroacetic acid salt), off-white solid.
• Another isomer compound is arbitrarily designated as (4 1 S,4 3 S,8S)-1 7 -(dimethylphosphoryl)-8-fluoro-10-methyl-2 5 -(trifluoromethyl)-1 1 H-3,5,10-tri Aza-1(3,6)-indole-2(4,2)-pyrimidine-4(1,3)-cyclopentanecycloundecane-11-one (4 1 S,4 3 S,8S )-1 7 -(dimethylphosphoryl)-8-fluoro-10-methyl-2 5 -(trifluoromethyl)-1 1 H-3,5,10-triaza-1(3,6)-indola-2(4,2 )-pyrimidina-4(1,3)-cyclopentanacycloundecaphan-11-one (Z77, 0.8g, yield 21%, trifluoroacetate salt), off-white solid.
• Step 12 To (4 1 S,4 3 S,8R)-1 7 -(dimethylphosphoryl)-8-fluoro-10-methyl-2 5 -(trifluoromethyl)-1 1 H -3,5,10-Triaza-1(3,6)-indole-2(4,2)-pyrimidine-4(1,3)-cyclopentanecycloundecane-11-one (Z76 , 220mg, 0.39mmol) in tetrahydrofuran (20mL) were successively added paraformaldehyde (176mg, 1.95mmol) and tetraisopropyl titanate (554mg, 1.95mmol), and the reaction solution was stirred at 70°C for 16h.
• paraformaldehyde 176mg, 1.95mmol
• tetraisopropyl titanate 554mg, 1.95mmol
• Step 13 To (4 1 S,4 3 S,8S)-1 7 -(dimethylphosphoryl)-8-fluoro-10-methyl-2 5 -(trifluoromethyl)-1 1 H -3,5,10-Triaza-1(3,6)-indole-2(4,2)-pyrimidine-4(1,3)-cyclopentanecycloundecane-11-one (Z77 , 200mg, 0.35mmol) in tetrahydrofuran (20mL) were successively added paraformaldehyde (158mg, 1.75mmol) and tetraisopropyl titanate (497mg, 1.75mmol), and the reaction solution was stirred at 70°C for 16h.
• paraformaldehyde 158mg, 1.75mmol
• tetraisopropyl titanate 497mg, 1.75mmol
• Step 1 To a solution of 7-bromo-1H-indole-6-carboxylate (5.0 g, 19.8 mmol) in N,N dimethylformamide (30 mL) at room temperature, add N-iodobutane Imide (6.7 g, 29.7 mmol). The reaction was stirred at 40°C for 2 hours under nitrogen protection. After the completion of the reaction as detected by LCMS, the reaction solution was added with saturated ammonium chloride solution (40 mL), and then extracted with ethyl acetate (40 mL ⁇ 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by filtration.
• Step 2 At room temperature, add 7-bromo-3-iodo-1H-indole-6-carboxylic acid methyl ester (6.5g, 17.2mmol), tris(dibenzylideneacetone) di Palladium (1.6g, 1.7mmol), 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (0.8g, 1.7mmol), pinacol borane (6.5g, 51.6mmol), tri Ethylamine (5.2 g, 51.6 mmol) and tetrahydrofuran (60 mL). Stir at 90°C for 15 hours under nitrogen protection.
• Step 3 Add 7-bromo-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl)-1H-indole-6-carboxylic acid methyl ester (2.4g, 6.3mmol), bistriphenylphosphine palladium dichloride (0.44g, 0.63mmol), potassium carbonate (1.7g, 12.6mmol) , tert-butyl ((1S, 3S)-3-((4-chloro-5-(trifluoromethyl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (2.4g, 6.3mmol) , a mixed solution of dioxane (30 mL) and water (6 mL).
• Step 4 At room temperature, methyl 7-bromo-3-(2-(((1S,3S)-3-((tert-butoxycarbonyl)amino)cyclopentyl)amino)-5-(trifluoromethyl Base) pyrimidin-4-yl)-1H-indole-6-carboxylate (1.4g, 2.3mmol) was dissolved in a mixed solution of hydrogen chloride/ethyl acetate (15mL, 4M) and ethyl acetate (10mL), React at 40°C for 16 hours.
• Step 5 At room temperature, methyl 3-(2-(((1S,3S)-3-aminocyclopentyl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)-7-bromo -1H-indole-6-carboxylate (500mg, 1.0mmol) in N,N-dimethylformamide (10mL) solution was added successively tert-butyl (4-iodobutyl) (methyl) aminomethyl Ester (344mg, 1.1mmol) and N,N-diisopropylethylamine (645mg, 5.0mmol), stirred at 70°C for 16 hours.
• Step 6 Methyl 7-bromo-3-(2-(((1S,3S)-3-((4-(tert-butoxycarbonyl)(methyl)amino)butyl)amino)cyclopentyl) Amino)-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylate (60mg, 0.09mmol) was dissolved in water (3mL), methanol (3mL) and tetrahydrofuran (3mL) Lithium hydroxide monohydrate (19mg, 0.45mmol) was added to the mixed system, and stirred at 40°C for 6 hours.
• Step 7 7-bromo-3-(2-(((1S,3S)-3-((4-((tert-butoxycarbonyl)(methyl)amino)butyl)amino)cyclopentyl)amino )-5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (60mg, 0.09mmol) was dissolved in methanol (5mL), then hydrochloric acid/methanol (5mL, 4M) was added , and stirred at 20°C for 16 hours.
• Step 8 To 7-bromo-3-(2-(((1S,3S)-3-((4-(methylamino)butyl)amino)cyclopentyl)amino)-5-(trifluoromethyl 2-(7-azabenzo Triazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (19 mg, 0.05 mmol) and N,N-diisopropylethylamine (32 mg, 0.25 mmol). The reaction was stirred at 60°C for 18 hours.
• Step 9 In a 50mL single-necked bottle, add (8S,11S)-20-bromo-17-methyl-3-(trifluoromethyl)-5,7,12,17,22,28-hexaaza Pentacyclic [17.4.3.1 2,6 .1 8,11 .0 21,24 ] twenty-eight-1(23),2(3),4,6(28),19(20),21(24) , 25-Heptaen-18-one (12mg, 0.022mmol, trifluoroacetate) in tetrahydrofuran (5mL) was added successively with paraformaldehyde (10mg, 0.11mmol) and tetraisopropyl titanate (31mg, 0.11 mmol), the reaction solution was stirred at 60°C for 14 hours, the reaction solution was lowered to 30°C, sodium cyanoborohydride (7 mg, 0.11 mmol) was added, and stirring was continued at 30°C for 4 hours.
• paraformaldehyde 10
• Step 10 At room temperature, sequentially add (8S,11S)-20-bromo-12,17-dimethyl-3-(trifluoromethyl)-5,7,12,17,22 into the microwave tube ,28-hexaazapentacyclo[17.4.3.1 2,6 .1 8,11 .0 21,24 ]28-1(23),2(3),4,6(28),19(20 ), 21(24), 25-heptaen-18-one (5mg, 0.009mmol), tetrakistriphenylphosphopalladium (2mg, 0.002mmol), zinc cyanide (3mg, 0.027mmol) and N,N-di Methylformamide (1 mL) solution. Microwave reaction at 150° C.
• Embodiment 34 compound Z85, the synthesis of Z86
• Step 1 At room temperature, add 4-methylbenzenesulfonic acid-7,7-difluoro-2,2,5-trimethyl-4-oxylidene-5-aza-3-oxanonyl-9 Add methylamine methanol solution (30% content, 20mL) and sodium iodide (60mg, 0.38mmol) to a methanol (10mL) solution of -yl ester (1.50g, 3.81mmol), and stir at 50°C for 14 hours. After LCMS detected that the reaction was complete, the reaction solution was concentrated to remove the solvent.
• Step 2 Dissolve (4,4-difluoro-2,7-diazaoct-2-yl)methanoic acid-2-methylpropan-2-yl ester (1.10g, 3.49mmol) in tetrahydrofuran (10mL ), then added ethyl trifluoroacetate (2.48g, 17.4mmol) and triethylamine (1.5mL, 10.5mmol) successively, and stirred at 50°C for 14 hours. After the reaction was detected by LCMS, ethyl acetate ( 30ml) and water (20mL), the organic phase was separated and concentrated.
• Step 3 Add (4,4,9,9,9-pentafluoro-7-methyl-8-oxyylidene-2,7-diazanon-2-yl)methanoic acid-2-methylpropane -2-yl ester (566mg, 1.63mmol) was dissolved in dichloromethane (5mL), then trifluoroacetic acid (1.0mL) was added to the reaction solution, and stirred at 25°C for 3 hours.
• Step 4 Dissolve N-[3,3-difluoro-4-(methylamino)butyl]-2,2,2-trifluoro-N-methylacetamide (399mg, 1.61mmol) in methanol ( 5mL), then add ⁇ [(3S)-1-oxyethylenecyclopent-3-yl]amino ⁇ methanoic acid benzyl ester (562mg, 2.41mmol), 4A molecular sieves (100mg), glacial acetic acid (2 drops) After stirring at 25°C for 2 hours, sodium cyanoborohydride (303 mg, 4.82 mmol) was added to the reaction solution, and stirring was continued at 25°C for 14 hours.
• Step 5 Add ⁇ [(3S)-1-(4,4,9,9,9-pentafluoro-7-methyl-8-oxyylidene-2,7-diazanon-2-yl) Cyclopent-3-yl]amino ⁇ methanoic acid benzyl ester (330mg, 2.21mmol) was dissolved in methanol (10mL), palladium carbon (30mg, 10% content, 55% water content), glacial acetic acid ( 30mg, 0.50mmol), replace the hydrogen, stir at 25°C for 2 hours, after TLC detects that the reaction is complete, filter and concentrate the organic phase to obtain the crude product as colorless oily N-(3,3-difluoro-4- ⁇ [(3S)- 3-Aminocyclopentyl](methyl)amino ⁇ butyl)-2,2,2-trifluoro-N-methylacetamide (310mg, yield 96%) can be directly used in the next reaction.
• LCMS m/z (ESI
• Step 6 Add N-(3,3-difluoro-4- ⁇ [(3S)-3-aminocyclopentyl](methyl)amino ⁇ butyl)-2,2,2-trifluoro-N- Methylacetamide (310mg, 0.94mmol) was dissolved in N,N-dimethylformamide (4ml), and 3-[2-chloro-5-(trifluoromethyl)pyrimidine-4- base]-7-[dimethyl(oxyethylene)- ⁇ 5 -methylphosphoryl]-1H-indole-6-carboxylic acid methyl ester (424mg, 0.98mmol), N,N-diisopropylethylamine (0.77mL, 4.68mmol), the reaction was stirred at 130°C for 5 hours under a nitrogen atmosphere, after the LCMS detected that the reaction was complete, water (10mL) was added to the reaction solution, then extracted with ethyl acetate (10mL ⁇ 2), and the combined organic layers , dried over anhydr
• Step 7 Add 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-(4,4,9,9,9- Pentafluoro-7-methyl-8-oxyylidene-2,7-diazanon-2-yl)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H -Methyl indole-6-carboxylate (125mg, 0.17mmol) was dissolved in a mixed solvent of water (1mL), methanol (3mL) and tetrahydrofuran (3mL), and lithium hydroxide (1 crystal water, 300mg, 7.09mmol ), stirred at 40°C for 14 hours.
• Step 8 To 7-[dimethyl(oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S,3S)-3-(4,4-difluoro-2,7 N,N -N,N-Diisopropylethylamine (37.7mg, 0.29mmol) and 2-(7-azabenzotriazole)-N,N,N were added in turn to dimethylformamide (4mL) solution ', N'-Tetramethylurea hexafluorophosphate (74mg, 0.20mmol), stirred at 60°C for 14 hours.
• dimethylformamide (4mL) solution ', N'-Tetramethylurea hexafluorophosphate 74mg, 0.20mmol
• Embodiment 35 compound Z87, the synthesis of Z88
• Step 2 Dissolve benzyl (4-fluoro-2,7-diazaoctyl-2-yl)carboxylate (1.50g, 5.59mmol) in dichloromethane (20mL), and then add 4-dimethylamino in sequence Pyridine (0.07g, 0.56mmol) and di-tert-butyl dicarbonate (1.46g, 6.71mmol) were stirred at 25°C for 14 hours. After the reaction was complete as detected by LCMS, the reaction solution was concentrated to remove the solvent.
• Step 3 Benzyl (8-fluoro-2,2,5-trimethyl-4-oxyylidene-5,10-diaza-3-oxaundecane-10-yl)methanoate (1.20g, 3.26mmol) was dissolved in methanol (20mL), and palladium on carbon (120mg, 10% content, 55% water content) was added to the reaction solution.
• Step 4 Dissolve (5-fluoro-2,7-diazaoct-2-yl)methanoic acid-2-methylpropan-2-yl ester (600mg, 2.56mmol) in methanol (15mL), and add ⁇ [(3S)-1-Oxylidenecyclopent-3-yl]amino ⁇ methanoic acid benzyl ester (597mg, 2.56mmol), glacial acetic acid (30mg, 0.50mmol), stirred at 25°C for 2 hours, and added to the reaction solution Sodium cyanoborohydride (483mg, 7.68mmol) was added, and stirring was continued at 25°C for 16 hours.
• Step 5 Dissolve 3-aminocyclopent-1-ol hydrochloride (10.0g, 72.7mmol) in dichloromethane (100mL), add benzyloxycarbonyl succinimide (21.7g , 87.2mmol), triethylamine (14.7g, 145mmol). Under nitrogen atmosphere, stir at 25°C for 14 hours, LCMS detects that the reaction is complete, add water (100mL) to the reaction solution, extract with dichloromethane (200mL ⁇ 3), wash the combined organic layer with saturated brine, dry over anhydrous sodium sulfate, and filter concentrate.
• Step 6 At -78°C, add dimethyl sulfoxide (1.69g, 21.7mmol) and oxalyl chloride (1.38g, 10.8mmol) to dichloromethane (20mL) successively, and stir the reaction solution at -78°C for 1h .
• Benzyl [(3-hydroxycyclopentyl)amino]methanoate (1.7g, 7.23mmol) was added to the reaction solution, and the reaction solution was continued for 3h, and triethylamine (3.66g, 36.1mmol) was added to the reaction solution.
• LCMS detected that the reaction was complete.
• Step 7 Add ⁇ 7-[(3S)-3- ⁇ [(benzyloxy)carbonyl]amino ⁇ cyclopentyl]-5-fluoro-2,7-diazaoct-2-yl ⁇ methane -2-Methyl prop-2-yl ester (P1, 1.00g, 2.21mmol) was dissolved in methanol (20mL), palladium carbon (100mg, 10% content, 55% water content), glacial acetic acid ( 30.0mg, 0.50mmol), the reaction was stirred at 25°C for 14 hours under hydrogen atmosphere.
• Step 8 Add ⁇ 7-[(3S)-3-aminocyclopentyl]-5-fluoro-2,7-diazaoct-2-yl ⁇ methanoic acid-2-methylpropan-2-yl ester (0.75g, 90% purity, 2.13mmol) was dissolved in N,N-dimethylformamide (10mL), and N,N-diisopropylethylamine (1.37g10.63mmol), 3 -[2-Chloro-5-(trifluoromethyl)pyrimidin-4-yl]-7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-1H-indole-6-carboxylic acid methyl ester (1.01 g, 2.34 mmol).
• Step 9 Add 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S)-3-(8-fluoro-2,2,5-trimethyl -4-Oxylidene-5,10-diaza-3-oxaundecane-10-yl)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H -Methyl indole-6-carboxylate (1.01g, 1.42mmol) was dissolved in a mixed system of water (5mL), methanol (5mL) and tetrahydrofuran (5mL), then lithium hydroxide (1 crystal water, 0.30g , 7.09mmol), stirred at 40°C for 5 hours. After the completion of the reaction detected by LCMS, the reaction solution was directly used in the next reaction without any treatment.
• LCMS: m/z (ESI): 699.4[M+H] + , t R 0.9
• Step 10 Add hydrogen chloride/methanol solution (10 mL, 4M) to the reaction solution in step 7, and stir the system at 40° C. for 5 hours.
• the solvent was directly spin-dried, and the obtained crude product was prepared by reverse-phase HPLC (chromatographic column: Ultimate XB-C18, 50*250mm, 10um (PARP-02); mobile phase: acetonitrile-water (0.1% bicarbonate ammonium); mobile phase: 70mL/min; column temperature: 25°C) to obtain 7-[dimethyl(oxygenide)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S)-3- (4-Fluoro-2,7-diazaoct-2-yl)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (530mg , yield 62%) was white solid.
• LCMS m/z (ESI)
• Step 11 To 7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S)-3-(4-fluoro-2,7-diazapine -2-yl)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (100mg, 0.17mmol) of N,N-dimethylformaldehyde Add N,N-diisopropylethylamine (108mg, 0.84mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetra Methylurea hexafluorophosphate (254mg, 0.67mmol), the reaction solution was stirred at 60°C for 14h.
• (4 1 S)-1 7 - (dimethylphosphoryl)-7-fluoro-5,10-dimethyl-2 5 -(trifluoromethyl)-1 1 H-3,5,10-triaza-1(3,6)-indola-2(4,2)-pyrimidina -4(1,3)-cyclopentanacycloundecaphan-11-one) (Z88, 8.24 mg; trifluoroacetate, yield 8.34%).
• Embodiment 36 compound Z89, the synthesis of Z90
• Step 1 Add ⁇ 7-[(3S)-3- ⁇ [(benzyloxy)carbonyl]amino ⁇ cyclopentyl]-5-fluoro-2,7-diazaoct-2-yl ⁇ methane -2-Methyl prop-2-yl ester (P2, 530mg, 1.17mmol) was dissolved in methanol (10mL), then palladium carbon (53mg, 10% content, 55% water content), glacial acetic acid ( 30mg, 0.50mmol). Under hydrogen atmosphere, stir at 25°C for 14 hours.
• Step 2 ⁇ 7-[(3S)-3-aminocyclopentyl]-5-fluoro-2,7-diazaoct-2-yl ⁇ methanoic acid-2-methylpropan-2-yl ester (380mg, 90% purity, 1.08mmol) was dissolved in N,N-dimethylformamide (8mL), and N,N-diisopropylethylamine (696mg 5.39mmol), 3-[ 2-Chloro-5-(trifluoromethyl)pyrimidin-4-yl]-7-[dimethyl(oxygenidene)- ⁇ 5-methylphosphonyl]-1H-indole-6-carboxylic acid methyl ester (465mg , 1.08mmol).
• Step 3 7-[Dimethyl(oxygenide)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S)-3-(8-fluoro-2,2,5-trimethyl -4-Oxylidene-5,10-diaza-3-oxaundecane-10-yl)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H -Methyl indole-6-carboxylate (330mg, 0.46mmol) was dissolved in a mixed system of water (3mL), methanol (3mL) and tetrahydrofuran (3mL), and lithium hydroxide (1 crystal water, 97mg, 2.32mmol ), and the reaction was stirred at 40°C for 5 hours. After the completion of the reaction detected by LCMS, the reaction solution was directly used in the next reaction without any treatment.
• Step 4 Add hydrogen chloride/methanol solution (8 mL, 4M) to the reaction solution in Step 3, and stir the system at 40° C. for 5 hours.
• the solvent was directly spin-dried, and the obtained crude product was prepared by reverse-phase HPLC (chromatographic column: Ultimate XB-C18, 50*250mm, 10um (PARP-02); mobile phase: acetonitrile-water (0.1% carbonic acid ammonium hydrogen); flow rate: 70mL/min; column temperature: 25°C) to obtain 7-[dimethyl(oxygenide)- ⁇ 5-methylphosphoryl]-3-(2- ⁇ [(1S)-3- (4-Fluoro-2,7-diazaoct-2-yl)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (190mg , Yield 74%), white solid.
• LCMS m/z (m/z)
• Step five to 7-[dimethyl (oxygenidene)- ⁇ 5 -methylphosphoryl]-3-(2- ⁇ [(1S)-3-(4-fluoro-2,7-diazapine -2-yl)cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-indole-6-carboxylic acid (100mg, 0.17mmol) of N,N-dimethylformaldehyde Add N,N-diisopropylethylamine (172mg, 1.33mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetra Methylurea hexafluorophosphate (254mg, 0.67mmol), the reaction solution was stirred at 60°C for 14 hours.
• (4 1 S)-1 7 - (dimethylphosphoryl)-7-fluoro-5,10-dimethyl-2 5 -(trifluoromethyl)-1 1 H-3,5,10-triaza-1(3,6)-indola-2(4,2)-pyrimidina -4(1,3)-cyclopentanacycloundecaphan-11-one) (Z90, 15.0 mg; yield 15.18%).
• Step 1 In a 25mL microwave tube, add 6-chloro-1H-pyrrolo[2,3-b]pyridine (1.0g, 6.55mmol), N-methylpyrrolidone (15mL), zinc cyanide (0.77g , 6.55mmol), tetrakis(triphenylphosphine)palladium (0.76g, 0.66mmol) and zinc powder (0.21g, 3.28mmol), and microwaved at 150°C for 6 hours.
• Step 2 Add N-iodosuccinyl to a solution of 1H-pyrrolo[2,3-b]pyridine-6-carbonitrile (1.20g, 8.38mmol) in N,N dimethylformamide (25mL) Imine (2.26 g, 10.1 mmol). Stir at 45°C for 2 hours under nitrogen protection. After the reaction was complete as detected by LCMS, the reaction solution was added with water (50 mL), extracted with ethyl acetate (50 mL ⁇ 3), the combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by filtration.
• Step 3 Add triethylamine (1.76g , 17.4mmol), 4-dimethylaminopyridine (0.17g, 1.39mmol) and di-tert-butyl dicarbonate (2.28g, 10.4mmol). Stir at room temperature for 14 hours. After the completion of the reaction was detected by LCMS, water (50 mL) was added to the reaction solution, and then extracted with ethyl acetate (50 mL ⁇ 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by filtration.
• Step 4 Add 6-cyano-3-iodopyrrolo[2,3-b]pyridine-1-carboxylic acid-2-methylpropan-2-yl ester (1.90g, 5.15mmol ), tris(dibenzylideneacetone)dipalladium (470mg, 0.515mmol), tetrahydrofuran (20mL), 2-dicyclohexylphosphine-2′,4′,6′-triisopropylbiphenyl (242mg, 0.515 mmol), triethylamine (1.56 g, 15.4 mmol) and 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.98 g, 15.4 mmol).
• Step five 6-cyano-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborol-2-yl)pyrrolo[2,3 -b]
• pyridine-1-carboxylic acid-2-methylpropan-2-yl ester (1.30g, 3.52mmol) in tetrahydrofuran (13mL)
• ⁇ [(1S,3S)-3- ⁇ [4-chloro -5-(trifluoromethyl)pyrimidin-2-yl]amino ⁇ cyclopentyl]amino ⁇ methanoic acid-2-methylpropan-2-yl ester (1.34g, 3.52mmol)
• Step 6 To 6-cyano-3-(2- ⁇ [(1S,3S)-3-( ⁇ [(2-methylpropan-2-yl)oxy]carbonyl ⁇ amino)cyclopentyl at room temperature ]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)pyrrolo[2,3-b]pyridine-1-carboxylic acid-2-methylprop-2-yl ester (700mg, 1.20mmol) Trifluoroacetic acid (2 mL) was added to the dichloromethane (10 mL) solution, and the reaction solution was reacted at 25° C. for 2 hours.
• Step 7 To 6-cyano-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)pyrrolo [2,3-b]Pyridine-1-carboxylic acid-2-methylpropan-2-yl ester (300mg, 0.774mmol) in N-methylpyrrolidone (5mL) was added successively (1-iodo-6-nitro Hetero-3-oxahept-6-yl)-2-methylpropan-2-yl methane (255mg, 0.77mmol) and N,N-diisopropylethylamine (0.384mL, 2.32mmol), The reaction was stirred at 80°C for 16 hours.
• Step 8 To (1- ⁇ [(1S,3S)-3- ⁇ [4-(6-cyano-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-( Trifluoromethyl)pyrimidin-2-yl]amino ⁇ cyclopentyl]amino ⁇ -6-aza-3-oxahept-6-yl)methanoic acid-2-methylpropan-2-yl ester (130mg , 0.221mmol) in methanol (5mL), add paraformaldehyde (100mg, 1.11mmol), acetic acid (0.1mL), 4A molecular sieves (130mg), and stir at 25°C for 2 hours under nitrogen protection.
• Step 9 Adding ⁇ 8-[(1S,3S)-3- ⁇ [4-(6-cyano-1H-pyrrolo[2,3-b]pyridin-3-yl)-5-(tri Fluoromethyl)pyrimidin-2-yl]amino ⁇ cyclopentyl]-2,8-diaza-5-oxanon-2-yl ⁇ methanoic acid-2-methylpropan-2-yl ester (100mg ,0.167mmol) of acetic acid (4mL) was added concentrated hydrochloric acid (2mL), and the reaction solution was stirred at 120°C for 16 hours.
• Step 10 To 3-(2- ⁇ [(1S,3S)-3-(2,8-diaza-5-oxanon-2-yl)cyclopentyl]amino ⁇ -5-(trifluoro Methyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridine-6-carboxylic acid (15mg, 0.029mmol) in N,N-dimethylformamide (3mL) solution, add in sequence N,N-Diisopropylethylamine (0.048mL, 0.29mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (54.7mg, 0.144mmol), the reaction solution was stirred at 60°C for 4h.
• Step 1 At room temperature, 3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)-1H-pyrrolo[ 2,3-b]pyridine-6-carbonitrile (220mg, 0.57mmol) in N-methylpyrrolidone (5mL), add [(4-iodobutyl)(methyl)amino]methane acid-2- Methylpropan-2-yl ester (178 mg, 0.57 mmol) and N,N-diisopropylethylamine (147 mg, 1.14 mmol).
• Step 2 At room temperature, to [(4- ⁇ [(1S,3S)-3- ⁇ [4-(6-cyano-1H-pyrrolo[2,3-b]pyridin-3-yl)-5 -(trifluoromethyl)pyrimidin-2-yl]amino ⁇ cyclopentyl]amino ⁇ butyl)(methyl)amino]methanoic acid-2-methylpropan-2-yl ester (120mg, 0.21mmol) To methanol (6 mL) solution, paraformaldehyde (94.0 mg, 1.05 mmol), acetic acid (30.0 mg), 4A molecular sieves (120 mg) were added, and the reaction was stirred at 25° C. for 2.0 hours under nitrogen protection.
• Step 3 At room temperature, to ⁇ 7-[(1S,3S)-3- ⁇ [4-(6-cyano-1H-pyrrolo[2,3-b]pyridin-3-yl)-5- (Trifluoromethyl)pyrimidin-2-yl]amino ⁇ cyclopentyl]-2,7-diazaoct-2-yl ⁇ methanoic acid-2-methylpropan-2-yl ester (110mg, 0.19mmol ) in acetic acid (4 mL) was added concentrated hydrochloric acid (2 mL), and stirred at 120°C for 14 hours.
• Step 4 To 3-(2- ⁇ [(1S,3S)-3- ⁇ [4-(methylamino)butyl]amino ⁇ cyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidine- 4-yl)-1H-pyrrolo[2,3-b]pyridine-6-carboxylic acid (40.0mg, 0.08mmol) in N,N-dimethylformamide (5mL) solution, add N,N- Diisopropylethylamine (51.0mg, 0.40mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (36.0mg, 0.10 mmol), and the reaction solution was stirred at 60° C.
• Step 1 At room temperature, to 6-cyano-3-(2- ⁇ [(1S,3S)-3-aminocyclopentyl]amino ⁇ -5-(trifluoromethyl)pyrimidin-4-yl)pyrrole And[2,3-b]pyridine-1-carboxylic acid-2-methylpropan-2-yl ester (2.0g, 5.16mmol) in N-methylpyrrolidone (20mL) solution, add 4-methylbenzene Sulfonic acid-7,7-difluoro-2,2,5-trimethyl-4-oxylidene-5-aza-3-oxanon-9-yl ester (2.44g, 6.20mmol), iodine Sodium chloride (0.15g, 1.03mmol) and N,N-diisopropylethylamine (2.56mL, 15.49mmol), the reaction was stirred at 80°C for 16 hours.
• Step 2 At room temperature, to [(2,2-difluoro-4- ⁇ [(1S,3S)-3- ⁇ [4-(6-cyano-1H-pyrrolo[2,3-b]pyridine -3-yl)-5-(trifluoromethyl)pyrimidin-2-yl]amino ⁇ cyclopentyl]amino ⁇ butyl)(methyl)amino]methanoate-2-methylpropan-2-yl ester (480mg, 0.789mmol) in methanol (5mL), add paraformaldehyde (118mg, 3.95mmol), acetic acid (0.1mL), 4A molecular sieves (480mg), and react under nitrogen protection at 25°C and stir for 2.0 hours.
• paraformaldehyde 118mg, 3.95mmol
• acetic acid 0.1mL
• 4A molecular sieves 480mg
• Step 3 At room temperature, to ⁇ 7-[(1S,3S)-3- ⁇ [4-(6-cyano-1H-pyrrolo[2,3-b]pyridin-3-yl)-5- (Trifluoromethyl)pyrimidin-2-yl]amino ⁇ cyclopentyl]-4,4-difluoro-2,7-diazaoct-2-yl ⁇ methanoic acid-2-methylpropan-2- Concentrated hydrochloric acid (1 mL) was added to a solution of base ester (330 mg, 0.53 mmol) in acetic acid (2 mL), and the reaction solution was stirred at 120°C for 5 hours.
• Step 4 To 3-(2- ⁇ [(1S,3S)-3-(5,5-difluoro-2,7-diazaoctyl-2-yl)cyclopentyl]amino ⁇ -5-( Trifluoromethyl)pyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridine-6-carboxylic acid (100mg, 0.19mmol) in N,N-dimethylformamide (2mL) solution, Add N,N-diisopropylethylamine (0.15mL, 0.92mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluoro Phosphate ester (280mg, 0.74mmol), the reaction solution was stirred at 60°C for 16 hours.
• Test Example 1 In vitro inhibition experiment of CDK kinase activity
• CDK2/Cyclin E1 was purchased from Byator; batch number: 20150228-BP469/477/691; CDK7/Cyclin H/MAT1 was purchased from Biotel; batch number: 20190326-BP487/492/479; CDK9/Cyclin T1 was purchased from Biotel Obtained; batch number: 20200727-BP488/792/691; CDK12/Cyclin K was purchased from Biotech; batch number: 20200526-BP1642/1648/691; MES was purchased from BioRoYee; batch number: 67GR9637; BSA was purchased from Aladdin ; Batch number: H1601024; ATP was purchased from VWR; Batch number: 97061-226; EDTA was purchased from Sinopharm Group; Batch number: 20200521;
• This experiment is used to determine the inhibitory effect of compounds on the kinase activity of CDK2, CDK7, CDK9 and CDK12.
• the kinase reaction carried out in the present invention is assayed in a 384-well plate, the final assay volume is 16 ⁇ l, and the reaction temperature is 27°C.
• Kinase concentrations were determined by optimization experiments.
• the specific experimental process is as follows:
• Kinase solution (CDK2/Cyclin E1): Kinase was diluted in assay buffer (20 mM MES pH 6.75, 0.01% Tween 20, 0.05 mg/mL BSA, 2 mM MgCl 2 ) to obtain a corresponding 2.4 ⁇ concentration enzyme solution.
• Kinase solution (CDK7/Cyclin H/MAT1): Kinase was diluted in assay buffer (20 mM MES pH 6.75, 0.01% Tween 20, 0.05 mg/mL BSA, 6 mM MgCl 2 ) to obtain a corresponding 2.4 ⁇ concentration enzyme solution.
• Kinase solution (CDK9/Cyclin T1): Kinase was diluted in assay buffer (20 mM MES pH 6.75, 0.01% Tween 20, 0.05 mg/mL BSA, 10 mM MgCl 2 ) to obtain a corresponding 2.4 ⁇ concentration enzyme solution.
• Kinase solution (CDK12/Cyclin K): Kinase was diluted in assay buffer (80 mM MES pH 6.5, 0.01% Tween 20, 0.05 mg/mL BSA, 10 mM MgCl 2 ) to obtain a corresponding 2.4 ⁇ concentration enzyme solution.
• Compound solution configuration the compound was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 10 mM. When used, the compound was diluted with DMSO to 10 concentration gradients from 25 nM to 500 ⁇ M, and diluted 8.3 times in ultrapure water to obtain Compound solution at 6X concentration.
• DMSO dimethyl sulfoxide
• Polypeptide substrate and ATP solution configuration The polypeptide substrate and ATP were diluted in the assay buffer to obtain a mixed solution of the polypeptide substrate and ATP at a concentration of 2.4 ⁇ .
• ATP 100 ⁇ M
• polypeptide substrate 5-FAM-YSPTSPSYSPTSPSYSPTSPSKKKK
• CDK2/Cyclin E1 0.5 nM
• ATP is 50 ⁇ M
• polypeptide substrate (5-FAM-YSPTSPSYSPTSPSYSPTSPSKKKK) is 2 ⁇ M
• CDK7/Cyclin H/MAT1 is 3 nM;
• ATP 50 ⁇ M
• peptide substrate FITC-Ahx-GSRTPMY-NH2
• CDK9/Cyclin T1 8 nM
• the CDK7 kinase activity inhibition of the compound of the present invention has reached the lower limit of the experiment, and the IC50 cannot accurately reflect the kinase activity inhibitory ability of the compound.
• SPR surface plasmon resonance
• CDK7/Cyclin H was purchased from Biotel; lot number: 20200309-BP487/492; MES buffer was purchased from BioRoYee; lot number: 67GR9637; CM5 sensor chip was purchased from Cytiva; lot number: 10305527; HEPES buffer was purchased from Cytiva; 32349.
• CDK7/Cyclin H dimer at a concentration of 50 ⁇ g/mL was amino-coupled to the CM5 sensor chip at a flow rate of 5 ⁇ L/min.
• the target protein is immobilized on the chip channel within 600 seconds, and generally reaches 7000-10000 response values.
• Compounds were serially diluted 2-fold in 5 steps to a concentration range of 0.6-10 nM in 10 mM HEPES buffer pH 7.4 with 150 mM NaCl, 0.05% surfactant P20 and 2% DMSO. Each compound concentration cycle was run at 100 ⁇ L/min, 180 s contact time and 1800 s dissociation time.
• K D (M) k off (s ⁇ 1 )/K on (M ⁇ 1 s ⁇ 1 ); wherein, K on (ka) is the association rate; K off (kd) is the dissociation rate; s ⁇ 1 ( per second) and M -1 s -1 (per mole per second) are the units of k off and k on .
• Table 2 The results of some example compounds are shown in Table 2.
• HCC70 was purchased from ATCC; Product No.: CRL-2315; RPMI1640 was purchased from Gibco; Product No.: 11875-093; Pancreatin (with EDTA) was purchased from Gibco; -Glo was purchased from Promega; article number: G7573; DMSO was purchased from VWR AMRESCO; article number: 0231-500ML; Staurosporine was purchased from Selleck; article number: S1421; ;Model: Envison.
• HCC70 is human breast ductal carcinoma cell cultured in RPMI-1640 medium containing 10% FBS. The cells in the logarithmic growth phase were taken, digested with trypsin-EDTA, collected and counted, and seeded 2000 HCC70 cells/well in a 384-well cell plate, and cultured overnight in 5% CO 2 .
• the cell group treated with DMSO was used as the experimental control (control), and the cell group treated with 5 ⁇ M Staurosporine was used as the blank control (blank).
• control the experimental control
• 5 ⁇ M Staurosporine was used as the blank control (blank).
• the results show that the compound of the present invention has a strong inhibitory effect on the proliferation of HCC70 human breast ductal carcinoma cells.
• the IC 50 of some compounds is lower than 1000nM or 500nM; even lower than 100nM or 50nM.
• the results of some exemplary compounds are shown in Table 3 below.
• OVCAR-3 was purchased from ATCC; Product No.: HTB-161; RPMI1640 was purchased from Gibco; Product No.: 11875-093; Pancreatin (with EDTA) was purchased from Gibco; ; CellTiter-Glo was purchased from Promega; Product No.: G7573; Bovine insulin was purchased from absin; Product No.: abs9169; DMSO was purchased from VWR AMRESCO; Model: NC-200; microplate reader purchased from PerkinElmer; model: Envison.
• OVCAR-3 is a human ovarian adenocarcinoma cell cultured in RPMI-1640 medium containing 20% FBS and 0.01mg/mL Bovine insulin.
• the cells in the logarithmic growth phase were taken, digested with trypsin-EDTA, collected and counted, and seeded 2000 OVCAR-3 cells/well in a 384-well cell plate, and cultured overnight in 5% CO 2 .
• Use DMSO to prepare 1000X compound stock solution with 3.16 times gradient concentration, dilute 100 times to 10X compound stock solution with medium, add 10X compound stock solution to each cell culture well on the second day after cell seeding, and the final concentration is 1X with 0.1% DMSO.
• the cell group treated with DMSO was used as the experimental control (control), and the cell group treated with 5 ⁇ M Staurosporine was used as the blank control (blank).
• control the experimental control
• 5 ⁇ M Staurosporine was used as the blank control (blank).
• the IC 50 of some compounds is lower than 1000nM or 500nM; even lower than 100nM or 50nM.
• the results of some exemplary compounds are shown in Table 4 below.
• Test Example 5 In vivo pharmacokinetic experiment of the compound of the present invention
• mice male Beagle dogs, 8-11kg, were purchased from Jiangsu Masi Biotechnology Co., Ltd. Three dogs were given intravenous injection of 0.2 mg/kg or 1.0 mg/kg of the compound to be tested (solvent: 2% DMSO+10% Kolliphor HS 15+88% (6.818% hydroxypropyl- ⁇ -cyclodextrin aqueous solution), 0.2mg/mL or 1.0mg/mL) and 3 dogs were given oral administration of 2mg/kg (solvent: 5% DMSO+95% (10% hydroxypropyl- ⁇ -cyclodextrin saline solution), 0.4mg/ mL).

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