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CN104230960A - Four-ring anaplastic lymphoma kinase inhibitor - Google Patents

Four-ring anaplastic lymphoma kinase inhibitor Download PDF

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Publication number
CN104230960A
CN104230960A CN201410157687.XA CN201410157687A CN104230960A CN 104230960 A CN104230960 A CN 104230960A CN 201410157687 A CN201410157687 A CN 201410157687A CN 104230960 A CN104230960 A CN 104230960A
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group
membered
radical
hydrogen
alkoxy
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CN104230960B (en
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吴永谦
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Xuanzhu Biopharmaceutical Co Ltd
Shandong Xuanzhu Pharma Co Ltd
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Shandong Xuanzhu Pharma Co Ltd
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    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a four-ring anaplastic lymphoma kinase inhibitor as shown in a general formula (I), or stereisomers, or pharmaceutically acceptable salts, esters or solvates of the inhibitor, wherein A1, A2, A3, M, X, Y, Q, R4, R5, R6, R7 and n are defined in the specification. The invention also relates to a preparation method of these compounds, pharmaceutical preparations and pharmaceutical compositions containing these compounds, and application of the compound or the stereisomers, or the pharmaceutically acceptable salts, esters or solvates of the compound in preparation of medicines for treating and/or preventing ALK-mediated cancer related diseases.

Description

Tetrafused cyclic anaplastic lymphoma kinase inhibitors
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a tetracyclic anaplastic lymphoma kinase inhibitor or a stereoisomer thereof, or a pharmaceutically acceptable salt, an ester or a solvate thereof, a preparation method of the compounds, a pharmaceutical preparation and a pharmaceutical composition containing the compounds, and application of the compounds or the stereoisomer thereof, or the pharmaceutically acceptable salt, the ester or the solvate thereof in preparation of medicines for treating and/or preventing cancer-related diseases mediated by ALK.
Background
Anaplastic Lymphoma Kinase (ALK) is a member of the receptor tyrosine kinase family, and can recruit downstream proteins through autophosphorylation, thereby expressing specific genes and regulating cellular metabolism and growth. Anaplastic lymphoma kinase was first found in Anaplastic Large Cell Lymphoma (ALCL) and was later found to be highly expressed in non-small cell lung cancer (NSCLC).
The micromolecule inhibitor of ALK can influence the growth of tumor cells and play a role in resisting tumors, but a large number of clinical trials prove that the first generation ALK inhibitor Crizotinib is easy to generate drug resistance, so the second generation ALK inhibitor which has good curative effect on patients with drug resistance to Crizotinib is designed and screened, and the clinical significance is remarkable.
Currently known ALK inhibitors include Crizotinib (Perey), LDK378 (Novartis), AZD-3463 (AstraZeneca), and the like.
Therefore, a new compound structure different from the Crizotinib is searched through compound structure modification, the drug resistance and the physicochemical property of the compound are improved, the drug forming property is improved, the bioavailability of the compound is improved, and a small molecular inhibitor active on ALK mutation is searched, so that the compound has important significance for treating diseases caused by ALK mutation clinically.
Disclosure of Invention
The invention aims at developing a small molecule inhibitor aiming at ALK, and provides a tetra-fused cyclic anaplastic lymphoma kinase inhibitor with good effect on treating and/or preventing ALK-mediated cancer-related diseases. The specific technical scheme is as follows:
1. a compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof:
wherein,
A1is selected from C-R1Or N;
A2is selected from C-R2Or N;
A3is selected from C-R3Or N, and A1、A2And A3Not simultaneously selected from N;
R1and R3Each independently selected from hydrogen, hydroxyl, carboxyl, amino, nitro, sulfonyl, halogen atom, cyano, C1-6Alkyl radical, C1-6Alkoxy group, (C)1-6Alkyl radical)2Amino group, C2-6Alkenyl radical, C2-6Alkynyl or 3-14 membered cycloalkyl;
R2selected from hydrogen, cyano, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylthio, 3-to 14-membered heterocyclyloxy, C2-6Alkenyl radical, C2-6Alkynyl, 3-14 membered cycloalkyl, 3-14 membered heterocyclyl, 5-15 membered heteroaryl or 6-14 membered aryl;
q is selected from 3-8 membered heterocyclic group, 6-14 membered heterocyclic group, 6-12 membered bridged ring group or 6-12 membered spiro ring group;
R4selected from the following groups:
(1) hydrogen, amino, hydroxy, nitro, cyano, halogen atom, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylamino radical, (C)1-6Alkyl radical)2Amino group, C1-6Alkylaminocarbonyl, hydroxy C1-6Alkyl, hydroxy C1-6Alkylamino, halogeno C1-6Alkyl radical, C1-6Alkylsulfonyl radical, C1-6Alkylsulfonylamino, aminosulfonyl, aminosulfonylamino, C2-6Alkenyl or C2-6An alkynyl group,
(2) a 6-12 membered bridged ring group, a 6-12 membered spiro ring group, a 3-8 membered heterocyclic group or a 6-14 membered heterocyclic group optionally substituted with a substituent selected from the group consisting of amino group, hydroxyl group, nitro group, halogen atom, carboxyl group, C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl radical, C2-6Alkynyl, 3-to 8-membered heterocyclic group or 3-to 8-membered cycloalkyl;
n is selected from 0, 1,2,3,4 or 5, and when n is 0, R is4When n is not less than 2, R4Which may be the same or different from each other,
and when Q is selected from 3-8 membered heterocyclic group, R4Can not be selected from 3-8 membered heterocyclic groups;
R5and R6Each independently selected from hydrogen, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy, hydroxy C1-6Alkyl radical, C2-6Alkenyl or C2-6An alkynyl group,
or R5And R6Are linked to each other to form a 3-to 14-membered heterocyclic group or a 3-to 14-membered cycloalkyl group together with the carbon atom to which they are linked;
R7selected from hydrogen, cyano, nitro, hydroxy, amino, amidino, sulfonyl, C1-6Alkyl radical, C1-6Alkoxy group, halogen atom, 3-to 14-membered cycloalkyl group, C2-6Alkenyl or C2-6Alkynyl, said C1-6Alkyl radical, C1-6Alkoxy, 3-to 14-membered cycloalkyl, C2-6Alkenyl and C2-6Alkynyl groups may be independently optionally substituted with the following substituents: hydroxyl, carboxyl, amino, cyano, halogen atom, nitro or 3-14-membered heterocyclic group;
m is selected from O, S or N-R8，R8Selected from hydrogen, C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl or C2-6Alkynyl, said C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl and C2-6Alkynyl may optionally be substituted by C1-6Alkoxy substitution;
y is selected from N or C-R9；
X is selected from O, S or N-R9；
R9Selected from hydrogen, hydroxyl, carboxyl, amino, nitro, cyano, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy group, (C)1-6Alkyl radical)2Amino group, C2-6Alkenyl radical, C2-6Alkynyl or 3-14 membered cycloalkyl.
2. The compound shown in the general formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof has the preferable technical scheme that the compound is shown in the general formula (II):
wherein,
R1and R3Each independently selected from hydrogen, hydroxyl, carboxyl, amino, nitro, sulfonyl, halogen atom, cyano, C1-6Alkyl radical, C1-6Alkoxy group, (C)1-6Alkyl radical)2Amino group, C2-6Alkenyl radical, C2-6Alkynyl or 3-to 8-membered cycloalkyl;
R2selected from hydrogen, cyano, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylthio, 3-to 14-membered heterocyclyloxy, C2-6Alkenyl radical, C2-6Alkynyl, 3-to 8-membered cycloalkyl, 5-to 6-membered heterocyclyl or 5-to 6-membered heteroaryl;
q is selected from 4-7 membered heterocyclic group, 6-12 membered heterocyclic group, 7-10 membered bridged ring group or 6-12 membered spiro ring group;
R4selected from the following groups:
(1) hydrogen, amino, hydroxy, nitro, cyano, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino radical, (C)1-4Alkyl radical)2Amino group, C1-4Alkylaminocarbonyl, hydroxy C1-4Alkyl, hydroxy C1-4Alkylamino, halogeno C1-4Alkyl radical, C1-4Alkylsulfonyl radical, C1-4Alkylsulfonylamino, aminosulfonyl, aminosulfonylamino, C2-6Alkenyl or C2-6An alkynyl group,
(2) 6-10 membered bridged ring group, 6-12 membered spiro ring group, 4-7 membered heterocyclic group or 6-12 membered heterocyclic ring optionally substituted with a substituent selected from amino group, hydroxyl group, halogen atom, C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl or C2-6An alkynyl group;
n is selected from 0, 1,2 or 3, and when n is 0, R4When n is not less than 2, R4Which may be the same or different from each other,
and when Q is selected from 4-7 membered heterocyclic group, R4Cannot be selected from 4-7 membered heterocyclic groups;
R5and R6Each independently selected from hydrogen, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy, hydroxy C1-6Alkyl radical, C2-6Alkenyl or C2-6An alkynyl group,
or R5And R6Are connected with each other to form a 5-to 10-membered heterocyclic group or a 3-to 8-membered cycloalkyl group together with the carbon atom to which they are connected;
R7selected from hydrogen, cyano, nitro, hydroxy, amino, amidino, sulfonyl, C1-6Alkyl radical, C1-6Alkoxy group, halogen atom, 3-to 8-membered cycloalkyl group, C2-6Alkenyl or C2-6Alkynyl, said C1-6Alkyl radical, C1-6Alkoxy, 3-to 8-membered cycloalkyl, C2-6Alkenyl and C2-6Alkynyl groups may be independently optionally substituted with the following substituents: hydroxyl, carboxyl, amino, cyano, halogen atom, nitro or 5-to 10-membered heterocyclic group;
m is selected from O, S or N-R8，R8Selected from hydrogen, C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl or C2-6Alkynyl, said C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl and C2-6Alkynyl may optionally be substituted by C1-6Alkoxy substitution;
y is selected from N or C-R9；
X is selected from S or N-R9；
R9Selected from hydrogen, hydroxy, carboxyl, amino, cyano, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy group, (C)1-6Alkyl radical)2Amino group, C2-6Alkenyl radical, C2-6Alkynyl or 3-to 8-membered cycloalkyl.
3. A compound according to claim 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof,
wherein,
R1and R3Each independently selected from hydrogen, hydroxy, amino, sulfonyl, halogen atom, C1-4Alkyl or C1-4An alkoxy group;
R2selected from hydrogen, cyano, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylthio, 5-to 10-membered heterocyclyloxy, C2-6Alkenyl radical, C2-6Alkynyl, 3-to 8-membered cycloalkyl, 5-to 6-membered heterocyclyl or 5-to 6-membered heteroaryl;
q is selected from 5-6 membered heterocyclic group, 6-10 membered heterocyclic group, 7-9 membered bridged ring group or 7-11 membered spiro ring group;
R4selected from the following groups:
(1) hydrogen, amino, hydroxy, nitro, cyano, halogen atom, C1-4Alkyl or C1-4An alkoxy group,
(2) a 7-to 10-membered bridged ring group, a 7-to 10-membered spiro ring group, a 5-to 6-membered heterocyclic group or a 6-to 10-membered heterocyclic group, which is optionally substituted with a substituent selected from the group consisting of an amino group, a hydroxyl group, a halogen atom, C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl or C2-6An alkynyl group;
n is selected from 0, 1,2 or 3, and when n is 0, R4When n is not less than 2, R4Which may be the same or different from each other,
and when Q is selected from 5-6 membered heterocyclic group, R4Can not be selected from 5-6 membered heterocyclic groups;
R5and R6Each independently selected from hydrogen, halogen atoms, C1-4Alkyl or C1-4An alkoxy group,
or R5And R6Are connected with each other to form a 5-to 6-membered heterocyclic group or a 3-to 6-membered cycloalkyl group together with the carbon atom to which they are connected;
R7selected from hydrogen, cyano, nitro, hydroxy, amino, amidino, sulfonyl, C1-6Alkyl radical, C1-6Alkoxy group, halogen atom, 3-to 6-membered cycloalkyl group, C2-6Alkenyl or C2-6An alkynyl group;
m is selected from O, S or N-R8，R8Selected from hydrogen, C1-4Alkyl or C1-4Alkoxy radical, said C1-4Alkyl radical, C1-4Alkoxy may optionally be substituted by C1-4Alkoxy substitution;
y is selected from N or C-R9；
X is selected from S or N-R9；
R9Selected from hydrogen, hydroxy, amino, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy or 3-to 8-membered cycloalkyl.
4. A compound according to claim 3 or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof,
wherein,
R1、R2and R3Are independently selected from hydrogen or C1-4An alkyl group;
q is selected from 5-6 membered heterocyclic group, 6-10 membered heterocyclic group, 7-9 membered bridged ring group or 7-11 membered spiro ring group;
R4selected from the following groups:
(1) hydrogen, amino or C1-4An alkyl group, a carboxyl group,
(2) 7-9 membered bridged ring group or 5-6 membered heterocyclic group;
n is selected from 0 or 1, and when n is 0, R4In the absence of the presence of the agent,
and when Q is selected from 5-6 membered heterocyclic group, R4Can not be selected from 5-6 membered heterocyclic groups;
R5and R6Are independently selected from hydrogen or C1-4An alkyl group;
R7selected from hydrogen, cyano, hydroxyl, amino, methyl, ethyl or chlorine atoms;
m is selected from N-R8，R8Selected from hydrogen or C1-4An alkyl group;
y is selected from C-R9，R9Selected from hydrogen, methyl, ethyl or n-propyl;
x is selected from S.
5. A compound according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof,
wherein,
q is selected from 4-7 membered heterocyclic group;
R4selected from 7-8 bridge ring groups;
n is selected from 1.
6. A compound according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof,
wherein,
q is selected from
R4Is selected from
n is selected from 1.
7. A compound according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof,
wherein,
q is selected from 7-8 membered bridged ring groups;
R4selected from hydrogen, amino, C1-4An alkyl group or a 4-to 7-membered heterocyclic group;
n is selected from 0 or 1, and when n is 0, R4Is absent.
8. A compound according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof,
wherein,
q is selected from
R4Selected from the following groups:
(1) hydrogen, amino, methyl;
(2)
n is selected from 0 or 1, and when n is 0, R4Is absent.
9. A compound according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof,
wherein,
q is selected from 6-10 membered heterocyclic group or 7-11 membered spiro ring group;
R4selected from hydrogen, amino or C1-4An alkyl group;
n is selected from 0 or 1, and when n is 0, R4Is absent.
10. A compound according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof,
wherein,
A1and A3Each independently selected from CH;
A2is selected from C-R2Wherein R is2Selected from hydrogen or C1-6An alkyl group;
R5and R6Each independently selected from hydrogen or C1-4An alkyl group;
R7selected from hydrogen or cyano;
m is selected from NH;
y is selected from CH;
x is selected from S.
11. A compound according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof, selected from:
12. the pharmaceutical composition of the compound according to any one of claims 1 to 11, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof, and one or more pharmaceutically acceptable carriers and/or diluents can be prepared into any pharmaceutically acceptable dosage form.
13. The pharmaceutical composition according to claim 12, further comprising one or more antineoplastic and immunosuppressive agents selected from the group consisting of methotrexate, capecitabine, gemcitabine, doxifluridine, pemetrexed disodium, pazopanib, imatinib, erlotinib, lapatinib, gefitinib, vandetanib, herceptin, bevacizumab, rituximab, trastuzumab, paclitaxel, vinorelbine, docetaxel, doxorubicin, hydroxycamptothecin, mitomycin, epirubicin, pirarubicin, bleomycin, letrozole, tamoxifen, fulvestrant, triptorelin, flutamide, leuprolide, anastrozole, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, nitrogen mustard, maflan, onconine, carboplatin, platinum, and platinum, Cisplatin, oxaliplatin, carboplatin, topotecan, camptothecin, topotecan, everolimus, sirolimus, carcinostat, 6-mercaptopurine, 6-thioguanine, azathioprine, rhzomorph D, daunorubicin, doxorubicin, mitoxantrone, bleomycin, plicamycin, or aminoglutethimide.
14. Use of a compound according to any one of claims 1 to 11, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof, for the manufacture of a medicament for the treatment and/or prevention of an ALK-mediated cancer-related disease selected from brain tumor, non-small cell lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, rectal cancer, hepatic cancer, hepatoblastoma, papillary renal cell tumor, head and neck squamous cell tumor, wilms' tumor, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, glioma, prostate cancer, thyroid cancer, female genital tract cancer, carcinoma in situ, lymphoma, neuroblastoma, neurofibromatosis, thyroid cancer, bone cancer, skin cancer, colon cancer, glioma, Testicular cancer, small cell lung cancer, gastrointestinal stromal tumor, prostate tumor, mast cell tumor, multiple myeloma, or melanoma.
Detailed Description
The "halogen atom" as used herein means a fluorine atom, chlorine atom, bromine atom, iodine atom, etc.
Said "C" of the present invention1-6Alkyl "denotes straight or branched alkyl having 1 to 6 carbon atoms, including for example" C1-4Alkyl group "," C1-3Alkyl "and the like, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 1, 2-dimethylpropyl and the like. "C" according to the invention1-4Alkyl "means" C "as defined above1-6Examples of the "alkyl group" are specific examples in which the number of carbon atoms is 1 to 4.
Said "C" of the present invention1-6Alkoxy "means" C "as defined above1-6Alkyl "a group bonded to another structure through an oxygen atom, such as methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy, 1-methylpropoxy, 2-methylpropoxy, 1-dimethylethoxy, pentyloxy, 1-methylbutoxy, 2-methylbutoxy, 3-methylbutoxy, 1-dimethylpropyloxy, 1, 2-dimethylpropyloxy, 2-dimethylpropyloxy, 1-ethylpropyloxy, hexyloxy, 1-methylpentyloxy, 2-methylpentyloxy, 3-methylpentyloxy, 4-methylpentyloxy, 1-dimethylbutyloxy, 1, 2-dimethylbutyloxy, 1, 3-dimethylbutyloxy, 2-dimethylbutyloxy, 1-methylpropyloxy, 1-dimethylbutyloxy, 2-dimethylbutyloxy, 1-methylpropyloxy, 1-dimethylbutyloxy, 1-methylpropyloxy, 2, 3-dimethylbutyloxy, 3-dimethylbutyloxy, 1-ethylbutoxy, 2-ethylbutoxy, 1, 2-trimethylpropoxy, 1,2, 2-trimethylpropoxy, 1-ethyl-1-methylpropyloxy, 1-ethyl-2-methylpropyloxy and the like. "C" according to the invention1-4Alkoxy "means" C "as defined above1-6Examples of "alkoxy groupSpecific examples of the carbon number of 1 to 4.
Said "C" of the present invention2-6Alkenyl "means a straight or branched chain or cyclic alkenyl group of 2 to 6 carbon atoms containing at least one double bond, including, for example," C2-4Alkenyl group "," C2-3Alkenyl group "," C3-6Cycloalkenyl "and the like, such as ethenyl, propenyl, 2-propenyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and the like. The double bond may optionally be cis and trans.
Said "C" of the present invention2-6Alkynyl refers to a straight or branched chain alkynyl group of 2-6 carbon atoms containing at least one triple bond, including, for example, "C2-6Alkynyl group "," C2-4Alkynyl group "," C2-3Alkynyl "and the like, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl and the like.
Said "C" of the present invention1-6Alkylamino radical and C1-6Alkylthio group "," C1-6Alkylaminocarbonyl group and C1-6Alkylsulfonyl group "," C1-6Alkylsulfonylamino "refers to the above-mentioned" C "respectively1-6Alkyl "a group attached to another structure through amino, thio, aminocarbonyl, sulfonyl, sulfonylamino. The term "C1-4Alkylsulfonyl "means" C "mentioned above1-4Alkyl "a group attached to another structure through a sulfonyl group.
The "hydroxy group C" of the present invention1-6Alkyl group "," halogeno C1-6Alkyl "means hydroxy and halogen atom substituted for the above" C "respectively1-6Alkyl "and groups attached to other structures through an alkyl group. The term "hydroxy C1-4Alkyl group "," halogeno C1-4Alkyl "means hydroxy and halogen atom substituted for the above" C "respectively1-4Alkyl "and groups attached to other structures through an alkyl group, wherein the" halogen atom "is as previously described.
The "according to the invention" (C)1-6Alkyl radical)2Amino "means that two atoms of the amino group which can be substituted by the above-mentioned" C "group1-6Alkyl groups "and groups attached to other structures through amino groups.
The 3-14 membered cycloalkyl refers to a cycloalkyl derived by removing a hydrogen atom from an alkane part with 3-14 carbon atoms, and includes 3-8 membered cycloalkyl and 6-14 membered fused cycloalkyl.
The 3-8 membered cycloalkyl group refers to a cyclic alkyl group derived from an alkane moiety of 3-8 carbon atoms by removing one hydrogen atom, and includes, for example, "3-6 membered cycloalkyl group", "3-5 membered cycloalkyl group", "5-6 membered cycloalkyl group", and the like, and examples thereof include, but are not limited to: cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, methylcyclopropane, dimethylcyclopropane, methylcyclobutane, dimethylcyclobutane, methylcyclopentane, dimethylcyclopentane, methylcyclohexane, dimethylcyclohexane, etc. The term "3-6-membered cycloalkyl" as used herein refers to a specific example of the "3-8-membered cycloalkyl" having 3 to 6 carbon atoms.
The 6-to 14-membered fused cycloalkyl group means a 6-to 14-membered cyclic group formed by two or more cyclic structures sharing two adjacent carbon atoms with each other, and examples thereof include, but are not limited to: bicyclo [3.1.0] hexanyl, bicyclo [4.1.0] heptanyl, bicyclo [2.2.0] hexanyl, bicyclo [3.2.0] heptanyl, bicyclo [4.2.0] octanyl, octahydropentanyl, octahydro-1H-indenyl, decahydronaphthyl, tetradecahydrophenanthryl, bicyclo [3.1.0] hex-2-enyl, bicyclo [4.1.0] hept-3-enyl, bicyclo [3.2.0] hept-3-enyl, bicyclo [4.2.0] oct-3-enyl, 1,2,3,3 a-tetrahydropentanyl, 2,3,3a,4,7,7 a-hexahydro-1H-indenyl, 1,2,3,4,4a,5,6,8 a-octahydronaphthyl, 1,2,4a,5,6,8 a-hexahydronaphthyl, 1,2,3,4,5, 8 a-hexahydronaphthyl, 10-decahydrophenanthryl and the like.
The term "heteroatom" as used herein means N, O, C (O), S, SO and/or SO2Etc., preferably N, O, S, more preferably N, O.
The 3-14-membered heterocyclic group refers to a 3-14-membered cyclic group containing one or more heteroatoms, and includes a 3-8-membered heterocyclic group and a 6-14-membered heterocyclic group.
The 3-to 8-membered heterocyclic group is a monocyclic heterocyclic group containing 3 to 8 ring atoms (wherein at least one hetero atom is contained). Specific examples include, but are not limited to, 2, 5-dihydrothienyl, 4, 5-dihydropyrazolyl, 3, 4-dihydro-2H-pyranyl, 5, 6-dihydro-4H-1, 3-oxazinyl, aziridinyl, azetidinyl, thietanyl, tetrahydrofuranyl, tetrahydropyrrolyl, imidazolidinyl, pyrazolidinyl, tetrahydrofuranyl, 1, 4-dioxanyl, 1, 3-dithianyl, morpholinyl, piperazinyl, and the like. The "3-to 6-membered heterocyclic group", "4-to 7-membered heterocyclic group" and "5-to 6-membered heterocyclic group" of the present invention refer to specific examples of the "3-to 8-membered heterocyclic group" in which the number of ring atoms is 3-to 6-membered, 4-to 7-membered and 5-to 6-membered.
The 6-14 membered heterocyclic group refers to a fused ring structure containing 6-14 ring atoms (wherein at least one heteroatom) and formed by connecting two or more ring structures sharing two adjacent atoms, such as a structure formed by a benzo 3-8 membered heterocyclic group, a structure formed by a 3-8 membered heterocyclic group and a 3-8 membered heterocyclic group, and the like, and specific examples include but are not limited to: and a group formed by substituting an optionally substituted hydrogen atom with an isocyclic structure.
The "6-12 membered heterocyclic group" and "6-10 membered heterocyclic group" in the present invention refer to specific examples of the "6-14 membered heterocyclic group" in which the number of ring atoms is 6-12 membered and 6-10 membered.
The 5-15-membered heteroaryl group refers to a cyclic aromatic group with 5-15 membered ring atoms and one or more heteroatoms, and comprises a 5-8-membered single heteroaryl group and an 8-15-membered thick heteroaryl group.
5-to 8-membered monoheteroaryl including, but not limited to, pyrrolyl, imidazolyl, pyrazolyl, 1,2, 3-triazolyl, 1,2, 4-triazolyl, pyridyl, furyl, thienyl, and the like,Azolyl radical, isoOxazolyl, thiazolyl, isothiazolyl, 1,2, 3-thiadiazolyl, 1,2, 4-thiadiazolyl, 1,3, 4-thiadiazolyl, 1,2,3-Oxadiazolyl, 1,2,4Oxadiazolyl, 1,2,5-Oxadiazolyl, 1,2, 3-triazinyl, 1,2, 4-triazinyl, tetrazolyl,Triazolyl, 2H-1,2Oxazinyl, 4H-1,2-Oxazinyl, 6H-1,2-Oxazinyl, 2H-1,3-Oxazinyl, 4H-1,3-Oxazinyl, 6H-1,3-Oxazinyl, 2H-1,4-Oxazinyl, 4H-1,4-Oxazinyl radical, iso-isomerOxazinyl, pyridazinyl, pyrimidinyl, pyrazinyl and the like;
an 8-15 membered fused heteroaryl group, which refers to a fused ring structure containing 8-15 ring atoms (wherein at least one heteroatom is present) joined by two or more heteroaryl rings sharing two adjacent atoms with each other, includes, but is not limited to, benzofuranyl, isobenzofuranyl, benzothienyl, indolyl, isoindolyl, quinolinyl, isoquinolinyl, indolizinyl, indazolyl, phthalazinyl, quinoxalinyl, quinazolinyl, benzodiazinyl, benzisoxazinylAzolyl, benzoAzinyl, benzimidazolyl, pyridopyridyl, pyrazolo [3,4-b]Pyridyl, purinyl, acridinyl, xanthenyl and the like. The preferable "8-to 12-membered thick heteroaryl", "8-to 10-membered heteroaryl" and "9-to 10-membered thick heteroaryl" refer to the specific examples in which the number of ring atoms in the "8-to 15-membered thick heteroaryl" is 8-to 12-membered, 8-to 10-membered and 9-to 10-membered, respectively.
The terms "5-to 10-membered heteroaryl group" and "5-to 6-membered heteroaryl group" refer to specific examples of the above "5-to 15-membered heteroaryl group" having 5-to 10-and 5-to 6-membered ring atoms.
The "6-to 14-membered aryl" refers to a monovalent moiety obtained by removing a hydrogen atom from a cyclic aromatic compound having 6-to 14-membered carbon atoms as a ring atom, and includes 6-to 8-membered aryl and 8-to 14-membered condensed ring aryl. The 6-to 8-membered aryl group includes phenyl, cyclooctatetraenyl and the like. The 8-to 14-membered fused ring aryl group is a fused ring group formed by two or more aromatic rings sharing two adjacent carbon atoms, and at least one ring is a cyclic group having an all unsaturated aromatic ring, and includes 8-to 14-membered all unsaturated fused ring carbon aryl groups such as naphthyl, anthryl, phenanthryl, etc., and also includes 8-to 14-membered partially saturated fused ring aryl groups such as benzo 3-to 8-membered cycloalkyl groups, specifically, 2, 3-dihydro-1H-indenyl, 1,2,3, 4-tetrahydronaphthyl, 1, 4-dihydronaphthyl, etc. The term "6-to 8-membered aryl" refers to a specific example of the above-mentioned "6-to 14-membered aryl" having 6 to 8 ring atoms.
The 6-12-membered bridged ring group refers to a structure which is formed by sharing two nonadjacent atoms by any two rings and contains 6-12 carbon atoms or/and heteroatoms. Examples thereof include "6 to 10-membered bridge ring group", "6 to 8-membered bridge ring group", "7 to 10-membered bridge ring group", "7 to 9-membered bridge ring group", "7 to 8-membered bridge ring group", "6 to 12-membered bridge heterocyclic group", "6 to 10-membered bridge heterocyclic group", "6 to 8-membered bridge heterocyclic group", "7 to 10-membered bridge heterocyclic group", "7 to 9-membered bridge heterocyclic group", "7 to 8-membered bridge heterocyclic group" and "8-membered bridge heterocyclic group". Examples include, but are not limited to:
and the like.
The 6-to 12-membered spiro ring group refers to a structure containing 6-to 12 carbon atoms or/and heteroatoms, wherein at least two rings share one atom. Examples thereof include "7-12-membered spiro ring group", "7-11-membered spiro ring group", "7-10-membered spiro ring group", "7-9-membered spiro ring group", "7-8-membered spiro ring group", "8-membered spiro ring", "6-12-membered spiro heterocyclic group", "7-11-membered spiro heterocyclic group", "7-10-membered spiro heterocyclic group", "7-9-membered spiro heterocyclic group", "7-8-membered spiro heterocyclic group" and "8-membered spiro heterocyclic group". Examples include, but are not limited to:
and the like.
The invention also provides three preparation methods of the compound, but not limited to the following methods, and the reaction equation is as follows:
the preparation method comprises the following steps:
the reaction steps are as follows:
step 1 preparation of intermediates 1,2, 7
Intermediates 1,2 and 7 were purchased or prepared separately.
Step 2 preparation of intermediate 3
Dissolving the intermediate 1 and the intermediate 2 in a proper amount of solvent (such as trifluoroacetic acid), heating and refluxing until the reaction is finished, cooling, removing the solvent, and separating by a proper method to obtain an intermediate 3.
Step 3 preparation of intermediate 4
Dissolving the intermediate 3 in a proper solvent, adding a proper amount of DDQ (dichlorodicyanobenzoquinone), heating at room temperature or heating until the reaction is finished, and separating by a proper method to obtain an intermediate 4.
Step 4 preparation of intermediate 5
And (3) uniformly mixing the intermediate 5 and a proper amount of pyridine hydrochloride or dissolving the mixture in a proper solvent, heating the mixture until the reaction is finished, and separating the mixture by a proper method to obtain the intermediate 5.
Step 5 preparation of intermediate 6
And (3) putting the intermediate 5 into a proper solvent, adding a proper amount of trifluoromethanesulfonic anhydride, stirring until the reaction is finished, and separating by a proper method to obtain an intermediate 6.
Step 6 preparation of intermediate 8
And (3) putting the intermediate 6 and a proper amount of the intermediate 7 into a proper solvent, heating until the reaction is finished, and separating and purifying by a proper method to obtain an intermediate 8.
Step 7 preparation of intermediate 9
The intermediate 8 and an appropriate amount of dess-martin reagent are added to an appropriate amount of the mixed solution (e.g., dichloromethane and dimethylsulfoxide) and stirred at room temperature (e.g., 2 hours). Intermediate 9 is isolated by a suitable method.
Step 8 preparation of Compounds of formula (I)
The intermediate 9 and indium chloride are dissolved in a proper amount of solvent (such as N, N-dimethylformamide) and stirred at room temperature (such as 3 hours) under the protection of nitrogen. Adding a proper amount of sodium cyanoborohydride, stirring at room temperature overnight, and separating by a proper method to obtain the compound of the general formula (I).
The preparation method 2 comprises the following steps:
the reaction steps are as follows:
step 1 preparation of intermediate 6
Refer to steps 1-6 of preparation method 1 above.
Step 2 preparation of intermediate 9
And dissolving the intermediate 7 and the intermediate 8 in a proper solvent, adding a proper amount of sodium cyanoborohydride, stirring at room temperature until the reaction is finished, and separating by a proper method to obtain an intermediate 9.
Step 3 preparation of Compounds of formula (I)
Intermediate 6 and intermediate 9 are dissolved in a suitable solvent, heated by microwave (e.g., 2 hours) with the addition of a suitable base (e.g., diisopropylethylamine), and isolated by a suitable method to provide the product.
The preparation method 3 comprises the following steps:
step 1 preparation of intermediate 2
Dissolving the intermediate 1 in a proper solvent (such as acetonitrile), adding N-bromosuccinimide at room temperature, stirring until the reaction is complete, adding water for quenching, and performing suction filtration to obtain an intermediate 2.
Step 2 preparation of intermediate 11
Refer to step 2 to step 8 of preparation method 1 described above.
Step 3 preparation of intermediate 12
The intermediate 11, triisopropylsilylacetylene, a base (e.g., cesium carbonate) and a suitable ligand (e.g., X-Phos dicyclohexylphosphonium-2 ',4',6' -triisopropylbiphenyl) and a palladium catalyst (e.g., bis (acetonitrile) palladium dichloride) are added to acetonitrile, heated (e.g., 80 ℃) to react under nitrogen protection, cooled (e.g., to room temperature), added with a solvent (e.g., ethyl acetate) and a saturated aqueous solution of sodium chloride, and the organic phases are separated and purified by a suitable method to give the intermediate 12.
Step 4 preparation of intermediate 13
Dissolving intermediate 12 in a suitable solvent (e.g. tetrahydrofuran), adding tetrabutylammonium fluoride, stirring at room temperature until the reaction is complete, adding water for quenching, extracting with an organic solvent (e.g. ethyl acetate), combining the organic phases, and purifying by suitable separation to give intermediate 13.
Step 5 preparation of the Compound of formula (II)
Dissolving the intermediate 13 in a proper solvent (such as methanol), adding a proper amount of palladium carbon, introducing hydrogen, stirring at room temperature until the reaction is complete, filtering the palladium carbon, concentrating the filtrate, and performing proper separation and purification to obtain the palladium-carbon composite material of the inventionGeneral formula (A)Ⅱ) Compound (I)。
In the reaction equation, A1、A2、A3、M、X、Y、Q、R1、R3、R4、R5、R6、R7And n is as defined hereinbefore, X' represents a halogen atom.
The pharmaceutically acceptable salt of any compound shown in the general formula (I) refers to a salt prepared from pharmaceutically acceptable and nontoxic alkali or acid, and comprises organic acid salt, inorganic acid salt, organic alkali salt and inorganic alkali salt. The organic acid salts include salts of formic acid, acetic acid, benzenesulfonic acid, benzoic acid, p-toluenesulfonic acid, camphorsulfonic acid, citric acid, methanesulfonic acid, ethanesulfonic acid, propanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, mucic acid, pamoic acid, pantothenic acid, succinic acid, tartaric acid, and the like. The inorganic acid salt includes salts of hydrobromic acid, hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid, and the like.
Organic base salts include primary, secondary and tertiary amines, and substituted amines include naturally occurring substituted amines, cyclic amines and basic ion exchange resins selected from the group consisting of betaine, caffeine, choline, N' -dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl-morpholine, N-ethylpiperidine, meglumine, glucosamine, hydrabamine, isopropylamine, methylglucamine, morpholine, piperazine, piperidine, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like. Natural amino acid salts such as glycine, alanine, valine, leucine, isoleucine, norleucine, tyrosine, cystine, cysteine, methionine, proline, hydroxyproline, histidine, ornithine, lysine, arginine, serine, and the like. Inorganic base salts include ammonium and salts of lithium, sodium, potassium, calcium, magnesium, zinc, barium, aluminum, iron, ketone, ferrous, manganese, manganous, and the like.
The present invention claims "stereoisomers" of the compounds of formula (I) refers to all stereoisomers produced when chiral centers, double bonds, etc. are present in the compounds of formula (I), including enantiomers, diastereomers, cis-trans isomers, tautomers, geometric isomers, epimers, and mixtures thereof, which are included in the scope of the present invention.
When any compound shown in the general formula (I) of the invention is synthesized to obtain a racemate, the required enantiomer-pure compound can be obtained by a chiral resolution method: chromatography with a chiral stationary phase (like high pressure preparative liquid phase, supercritical fluid chromatography) can be used. Chiral fillers include, but are not limited to: chiralcel OJ-H, Chiralpak AD-H, Chiralpak IA, and Chiralpak AS-H.
The "ester" of the compound of the general formula (I) of the present invention means an ester which can be formed by esterification with an alcohol in the presence of a carboxyl group in the compound of the formula (I), and which can be formed by esterification with an organic acid, an inorganic acid, an organic acid salt or the like in the presence of a hydroxyl group in the compound of the formula (I). The ester can be hydrolyzed in the presence of acid or alkali to generate corresponding acid or alcohol.
The compounds of formula (i) or stereoisomers thereof, or pharmaceutically acceptable salts or esters thereof, may be in the form of "solvates". Where the solvate is a hydrate, the hydration may be accomplished during the manufacturing process or may be gradual, taking advantage of the hygroscopic properties of the original anhydrous product.
The invention further claims a pharmaceutical composition which comprises any compound shown in the general formula (I) or a stereoisomer thereof, or pharmaceutically acceptable salt, ester or solvate thereof and one or more pharmaceutically acceptable carriers and/or diluents, and can be prepared into any pharmaceutically acceptable dosage form. Administered to a patient in need of such treatment by oral, parenteral, rectal, or pulmonary administration. For oral administration, it can be made into conventional solid preparations such as tablet, capsule, pill, granule, etc.; it can also be made into oral liquid, such as oral solution, oral suspension, syrup, etc. When the composition is formulated into oral preparations, appropriate filler, binder, disintegrating agent, lubricant, etc. can be added. For parenteral administration, it can be made into injection, including injection solution, sterile powder for injection and concentrated solution for injection. The injection can be prepared by conventional method in the existing pharmaceutical field, and can be prepared without adding additives or adding appropriate additives according to the properties of the medicine. For rectal administration, it can be made into suppository, etc. For pulmonary administration, it can be made into inhalant or spray.
The invention further claims pharmaceutical compositions comprising a compound of any of the general formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof, as defined above, together with one or more other antineoplastic agents and immunosuppressive agents. Such antineoplastic and immunosuppressive agents, such as antimetabolites, including but not limited to methotrexate, capecitabine, gemcitabine, doxifluridine, disodium pemetrexed; growth factor inhibitors, including but not limited to pazopanib, imatinib, erlotinib, lapatinib, gefitinib, vandetanib; antibodies, including but not limited to herceptin, bevacizumab; targeting classes, including but not limited to herceptin, bevacizumab, rituximab, trastuzumab; mitotic inhibitors, including but not limited to paclitaxel, vinorelbine, docetaxel, doxorubicin, hydroxycamptothecin, mitomycin, epirubicin, pirarubicin, bleomycin; antineoplastic hormones including, but not limited to, letrozole, tamoxifen, fulvestrant, triptorelin, flutamide, leuprolide, anastrozole; alkylating agents including, but not limited to, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, mechlorethamine, melphalan, and creme; the metal platins include, but are not limited to, carboplatin, cisplatin, oxaliplatin, and carboplatin; topoisomerase inhibitors including, but not limited to, topotecan, camptothecin, topotecan; immunosuppressive species including but not limited to everolimus, sirolimus, and temboticarb; purine analogs selected from the group consisting of 6-mercaptopurine, 6-thioguanine, azathioprine; antibiotics selected from the group consisting of rhzomycin D, daunorubicin, doxorubicin, mitoxantrone, and plicamycin; adrenocortical suppressants selected from aminoglutethimide.
The invention also provides application of the compound shown as the general formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt, an ester or a solvate thereof in preparing a medicament for treating and/or preventing ALK-mediated cancer-related diseases, wherein the cancer-related diseases are selected from brain tumor, lung cancer, non-small cell lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, rectal cancer, liver cancer, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, solid tumors, non-Hodgkin's lymphoma, thyroid cancer, female genital tract cancer, in situ cancer, lymphoma, histiocytoma, neurofibromatosis, bone cancer, skin cancer, colon cancer, testicular cancer, small cell lung cancer, gastrointestinal stromal tumor, prostate tumor, mast cell tumor, colon cancer, small cell lung cancer, gastrointestinal stromal tumor, colon cancer, Multiple myeloma, melanoma, glioma, astrocytoma, neuroblastoma, sarcoma, and the like.
The compound of the invention has the following advantages:
(1) the compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof has excellent inhibitory activity on ALK and NCI-H3122, Karpas-299 and NCI-H2228 cells;
(2) the compound of formula (I) or the stereoisomer or the pharmaceutically acceptable salt, ester or solvate thereof shows good pharmacokinetic property, and has lasting effect and high bioavailability;
(3) the compound of the invention has simple preparation process, high medicine purity, stable quality and easy large-scale industrial production.
The beneficial effects of the compounds of the present invention are further illustrated below by in vitro enzymatic inhibitory activity assays, but this should not be understood as meaning that the compounds of the present invention have only the following beneficial effects.
Experimental example 1 in vitro enzymatic Activity test of the Compound of the present invention
And (3) testing the sample: the chemical names and the preparation methods of the compounds 1 to 3, 5 to 11 and 13 of the present invention are shown in the preparation examples of the respective compounds. The abbreviations used in the following experiments have the following meanings:
DMSO, DMSO: dimethyl sulfoxide
DTT: dithiothreitol
SEB: enzyme catalyst buffer solution
ATP: adenosine triphosphate
ALK: anaplastic lymphoma kinase
SA-XL 665: streptavidin-labeled donors
2.5X, 5X, 10X wherein "X": multiple times
The experimental method comprises the following steps:
preparation of an ALK kinase buffer:
respectively taking a proper amount of MgCl with the mother liquor concentration of 1000mM2SEB (2500 nM), DTT (100 mM), 5 Xenzyme buffer were added to ultrapure water so that the final concentrations were: 5mM, 25nM, 1mM, 1 Xenzyme buffer, mixing, standby.
2.5 times of preparation of test solution:
1mM stock solution preparation of compound: respectively weighing appropriate amount of the compound (see the following table), adding appropriate amount of DMSO, dissolving, and mixing.
1mM stock solutions were each diluted with DMSO to prepare a 200. mu.M solution as a stock solution. The mother liquor is diluted three times step by DMSO to prepare a series of solutions with concentration, and then each concentration is diluted 80 times by ALK kinase buffer solution to prepare 2.5 times of test solution, the concentration is respectively: 2500nM, 833.33nM, 277.78nM, 92.59nM, 30.86nM, 10.29nM, 3.43nM, 1.14nM, 0.38nM, 0.13nM, 0.04 nM.
Various other reagent formulations:
the required 5 XALK kinase solution, 5 Xsubstrate solution and 5 XATP solution are prepared by ALK kinase buffer solution for standby.
ALK enzymatic reaction:
1) mu.L of the prepared 2.5 Xsample solution and 2. mu.L of the prepared 5 XALK kinase solution were added to corresponding wells of a 384-well plate, and incubated at 25 ℃ for 10 minutes.
2) mu.L of the prepared 5 Xsubstrate solution and 2. mu.L of the prepared 5 XATP solution were added to the corresponding wells, respectively, to initiate the enzymatic reaction, and incubated at 25 ℃ for 30 minutes.
And (3) enzymatic detection:
and (3) preparing SA-XL665 with a required concentration by using a detection buffer solution (detection buffer), then uniformly mixing the SA-XL665 with the tyrosine kinase antibody with the same volume, and respectively adding 10 mu L of the prepared antibody solution into corresponding holes to terminate the reaction. Incubate at 25 ℃ for 1 h.
The plate reader is 665nm/615 nm.
IC50: calculating the inhibition rate (%) = (maximum-sample ratio)/(maximum-minimum) × 100, and performing curve fitting by using Graph prism software to obtain IC50The value is obtained.
Maximum value: positive control without compound, minimum: negative control without enzyme.
Experimental results and conclusions:
TABLE 1 in vitro enzymatic inhibitory Activity of the Compounds of the invention
As can be seen from table 1, the compound of the present invention has good inhibitory activity on ALK kinase, and can be used for treating diseases related to kinase, particularly ALK kinase-mediated disorders or conditions, with significant clinical significance.
Experimental example 2 in vitro cell Activity experiment of the Compound of the present invention
And (3) testing the sample: the chemical names and the preparation methods of the compounds 1,2,3, 5,6 and 8 to 12 of the present invention are shown in the preparation examples of the respective compounds.
The abbreviations used in the following experiments have the following meanings:
rpm: rotate per minute
DMSO, DMSO: dimethyl sulfoxide
MTS: blue tetrazolium thiazole bromide
RPMI 1640: 1640 medium (RPMI: Roswell Park molar Institute)
500X, 1000X, 10X wherein "X": multiple times
Compound 1 experimental method:
NCI-H3122 and Karpas-299 cells:
(1) cell preparation:
using RPMI-1640 medium containing 10% fetal calf serum, 100U/ml penicillin, and 100mg/ml streptomycin in 5% CO2And culturing the cells in an incubator at 37 ℃ until the cells are 80% fused for later use.
(2) Inoculating cells:
digesting the cells with pancreatin, centrifuging at 1000rpm for 4min, removing the supernatant, resuspending with 10% fetal bovine serum-containing RPMI-1640 medium, adjusting the cell density, taking 90 μ L of the cell suspension, inoculating into a 96-well plate, and obtaining the final cell density Karpas-299: 1500/well; NCI-H3122: 2000 per well; then at 5% CO2And culturing in an incubator at 37 ℃ for 24 hours.
(3) Adding a test article:
(3.1) preparation method of Compound 1 solution: weighing a proper amount of compound 14.99mg, adding a proper amount of DMSO to dissolve, performing gradient dilution by using DMSO to prepare a series of mother solutions (1000 multiplied by test solution) with a series of concentrations, then diluting the mother solutions by 100 times by using a culture medium to obtain 10 multiplied by test solution, respectively taking 10 mu L of the solution, adding the solution into corresponding holes of a 96-well plate, and obtaining the final concentration of the test solution as follows: 10 μ M, 2.5 μ M, 625nM, 156nM, 39nM, 9.8nM, 2.5 nM.
(3.2) control well settings:
vehicle control: 0.1% DMSO.
Cell control: cells were seeded only, without compound addition.
Blank control: medium, instrument zero.
(3.3) placing the 96-well plate at 37 ℃ with 5% CO2Culturing for 72h in an incubator.
(4) And (3) detection:
(4.1) MTS detection method: suitable cells are: NCI-H3122 and Karpas-299
Firstly, the following steps are carried outThe reagents in the single-solution 96-well cell proliferation detection kit are placed at room temperature for 90 min.
② to 96-well plate to each test wellAQueous single solution reagent 20. mu.L.
Thirdly, putting 5% CO in 96-hole plate2And then cultured in an incubator at 37 ℃ for 2 hours.
And fourthly, setting the detection wavelength of the microplate reader to be 490nm, and reading the result.
(5) And (4) processing a result:
IC50and (3) calculating: cell viability (%) = (sample value-blank value)/(maximum value-blank value) × 100, curve fitting was performed using Graph prism software to obtain IC50The value is obtained.
Maximum value: cell control without compound plus vehicle alone, blank: blank control values.
(II) NCI-H2228 cells:
(1) cell preparation:
using RPMI-1640 medium containing 10% fetal bovine serum in 5% CO2And culturing the cells in an incubator at 37 ℃ until the cells are 80% fused for later use.
(2) Inoculating cells:
digesting the cells with pancreatin, centrifuging at 1000rpm for 4min, removing supernatant, resuspending with 10% fetal calf serum-containing RPMI-1640 medium, adjusting cell density to 2 × 104And (4) taking 100 mu L of the cell suspension, inoculating the cell suspension into a 96-well plate, and obtaining the final cell density: 2000 cells/well.
(3) Adding a test article:
(3.1) Compound 1 solution preparation: weighing 13.14mg of compound, adding a proper amount of DMSO, dissolving, uniformly mixing, and diluting the solution with DMSO in a gradient manner to obtain solutions with a series of concentrations for later use.
Adding 99 μ L of the culture medium into each well of a 96-well plate, and adding 1 μ L of the prepared solution with different concentrations into the corresponding wells, so that the final concentration of the compound 1 is: 10000nM,3333.33nM,1111.11nM,370.37nM,123.45nM,41.15nM,13.71nM,4.57nM,1.52nM,0.50 nM.
(3.2) placing 5% CO in the 96-well plate2And culturing for 96 hours in an incubator at 37 ℃.
(4) And (3) detection:
the CTG detection method comprises the following steps:
the culture medium in each well of the 96-well plate is removed, and 100. mu.L of new culture medium is added and the mixture is equilibrated at room temperature for 30 min.
② to 96-well plate to each test wellReagent 60 muL。
③ the 96 pore plate is shaken and mixed for 2min in a microplate oscillator in dark to crack the cells.
And fourthly, incubating the 96-well plate for 10min in a dark room temperature to stabilize the generated light signal value.
Reading the result by the microplate reader in a luminescence mode.
(5) And (4) processing a result:
curve fitting is carried out by adopting Graph prism software to obtain IC50The value is obtained.
Other part of the compounds of the invention experimental methods:
NCI-H3122, Karpas-299 cells:
(1) cell preparation:
using RPMI-1640 medium containing 10% fetal calf serum, 100U/ml penicillin, and 100mg/ml streptomycin in 5% CO2And culturing the cells in an incubator at 37 ℃ until the cells are 80% fused for later use.
(2) Inoculating cells:
digesting cells with pancreatin, centrifuging at 1000rpm for 4min, removing supernatant, resuspending with RPMI-1640 medium containing 2.5% fetal calf serum, adjusting cell density, taking 90 μ L of the cell suspension, and inoculating to 96-well plate to obtain final cell density of 3000 cells/well; then at 5% CO2And culturing in an incubator at 37 ℃ for 24 hours.
(3) Adding a test article:
(3.1) preparation of test solution
The first preparation method of the test solution comprises the following steps: weighing 22.93mg of a sample, adding a proper amount of DMSO (dimethyl sulfoxide), dissolving, diluting with DMSO gradient to prepare a series of mother solutions (200 multiplied by sample solutions) with concentration, diluting the mother solutions 20 times by using a culture medium to obtain 10 multiplied by sample solutions, respectively taking 10 mu L of the solutions, adding the solutions into corresponding holes of a 96-hole plate, and obtaining a sample 2 solution with the final concentration: 10 μ M, 2.5 μ M, 625nM, 156nM, 39nM, 9.8nM, 2.5 nM.
The preparation method of the test solution comprises the following steps: respectively weighing a proper amount of a sample (specifically weighing the sample amount shown in the following table), adding a proper amount of DMSO (dimethyl sulfoxide) for dissolving, respectively diluting with DMSO in a gradient manner to prepare a series of mother solutions (1000 multiplied by the sample solution) with concentration, respectively diluting the mother solutions by 100 times with a culture medium to obtain 10 multiplied by the sample solution, respectively taking 10 mu L of the solution, adding the solution into corresponding holes of a 96-hole plate, and obtaining the final concentration of each sample solution as follows: 10 μ M, 2.5 μ M, 625nM, 156nM, 39nM, 9.8nM, 2.5 nM.
(3.2) control well settings:
vehicle control: 0.1% DMSO or 0.2% DMSO, 0.5% DMSO.
Cell control: cells were seeded only, without compound addition.
Blank control: medium, instrument zero.
(3.3) placing the 96-well plate at 37 ℃ with 5% CO2Culturing for 72h in an incubator.
(4) And (3) detection:
the MTS detection method comprises the following steps:
firstly, the following steps are carried outThe reagents in the single-solution 96-well cell proliferation detection kit are placed at room temperature for 90 min.
② to 96-well plate to each test wellAQueous single solution reagent 20. mu.L.
Thirdly, putting 5% CO in 96-hole plate2And cultured in an incubator at 37 ℃ for 40 min.
And fourthly, setting the detection wavelength of the microplate reader to be 490nm, and reading the result.
(5) And (4) processing a result:
IC50and (3) calculating: cell viability (%) = (sample value-blank value)/(maximum value-blank value) × 100, curve fitting was performed using Graph prism software to obtain IC50The value is obtained.
Maximum value: cell control without compound plus vehicle alone, blank: blank control values.
(II) NCI-H2228 cells:
(1) cell preparation:
using RPMI-1640 medium containing 10% fetal bovine serum in 5% CO2And culturing the cells in an incubator at 37 ℃ until the cells are 80% fused for later use.
(2) Inoculating cells:
digesting the cells with pancreatin, centrifuging at 1000rpm for 4min, removing the supernatant, resuspending with RPMI-1640 medium containing 2.5% fetal calf serum (compounds 2, 9 and 12 with 10% fetal calf serum), adjusting the cell density to 2X 104And (4) taking 100 mu L of the cell suspension, inoculating the cell suspension into a 96-well plate, and obtaining the final cell density: 2000 cells/well.
(3) Adding a test article:
(3.1) preparing a test solution: respectively weighing appropriate amount of the test sample (see the following table), adding appropriate amount of DMSO, dissolving, mixing, and gradient diluting the solution with DMSO to obtain solutions with a series of concentrations.
Adding 99 μ L of culture medium into each well of a 96-well plate, and adding 1 μ L of the prepared solutions with different concentrations into the corresponding wells, so that the final concentration of the compound of the invention is: 10000nM, 2500nM, 625nM, 156.25nM, 39.06nM, 9.77nM, 2.44nM, 0.61nM, 0.15 nM. The final concentrations of the control drugs were: 9800nM, 2450nM, 612.5nM, 153.1nM, 38.28nM, 9.57nM, 2.39nM, 0.60nM, 0.15 nM.
(3.2) control well settings:
vehicle control: 0.5% DMSO.
Cell control: cells were seeded only, without compound addition.
Blank control: medium, instrument zero.
Paclitaxel control: cells were seeded with 1000nM paclitaxel and used as the lowest value control.
(3.3) placing 5% CO in the 96-well plate2And culturing for 96 hours in an incubator at 37 ℃.
(4) And (3) detection:
the CTG detection method comprises the following steps:
80 μ L of medium was removed from each well of the 96-well plate and allowed to equilibrate at room temperature for 30 min.
② to 96-well plate to each test wellReagent 60. mu.L.
③ the 96 pore plate is shaken and mixed for 2min in a microplate oscillator in dark to crack the cells.
And fourthly, incubating the 96-well plate for 10min in a dark room temperature to stabilize the generated light signal value.
Reading the result by the microplate reader in a luminescence mode.
(5) And (4) processing a result:
the first calculation method comprises the following steps: IC (integrated circuit)50And (3) calculating: cell inhibition ratio (%) = (OD)Solvent－ODCompound (I))/(ODSolvent－ODpositive) X 100, curve fitting with Graph prism software to obtain IC50The value is obtained.
ODSolvent: cell control, OD, without Compound plus vehicle alonepositive: cell control with 1000nM paclitaxel.
And a second calculation method comprises the following steps: IC (integrated circuit)50And (3) calculating: cell inhibition ratio (%) = (OD)Solvent－ODCompound (I))/(ODSolvent－ODNegative of) X 100, curve fitting with Graph prism software to obtain IC50The value is obtained.
ODSolvent: cell control, OD, without Compound plus vehicle aloneNegative of: media blank control
The experimental results are as follows:
TABLE 2 cytostatic Activity of Compounds of the invention
As can be seen from Table 2, the compounds of the invention have good inhibitory activity on cells NCI-H3122, Karpas-299 and NCI-H2228, can be used for treating ALK kinase-mediated disorders or conditions, and have significant clinical significance.
Experimental example 3 rat in vivo pharmacokinetic experiment of the Compound of the present invention
And (3) testing the sample: the chemical name and the preparation method of the compound 11 are shown in the preparation example of the compound 11.
The test animals were: male SD rats, 3/route of administration/test article; the weight of the rat used for the compound of the invention is 200-230 g/rat.
Preparing a test solution:
animal IV dosing group:
weighing 112.87mg of the compound, adding 101.46 mu L of DMSO, carrying out ultrasonic dissolution, adding 1.015mL of 40% HP-beta-CD (hydroxypropyl-beta-cyclodextrin), carrying out vortex mixing, carrying out heat preservation in a constant temperature water bath kettle at 50 ℃ for 20min, adding 3.957mL of sterilized water for injection, carrying out vortex mixing, and filtering to obtain the compound.
The preparation method of the 40% HP-beta-CD solution comprises the following steps: and adding 10mL of sterilized injection water into HP-beta-CD 4g, dissolving by ultrasonic, and mixing uniformly by vortex to obtain the compound.
Animal PO dosing group:
weighing 113.06mg of compound, adding 2% HPC +0.1% Twen 805.586mL, placing in a tissue grinder at a rotation speed of 600rpm/min, and dispersing uniformly to obtain the final product.
2% HPC +0.1% Tween80 solution preparation method: weighing 20g of hydroxypropyl cellulose (HPC), slowly adding into 1000mL of water, continuously stirring to fully dissolve, adding 1mL of Tween80, and mixing uniformly to obtain the final product.
Experimental methods
Administration the test solution was administered by intravenous bolus (iv) at a dose of 1mg/kg and a volume of 2 mL/kg; the test sample is administered by intragastric administration (po), the administration dose is 2mg/kg, and the administration volume is 4 mL/kg.
Blood sampling
Tail vein blood collection was performed 0.083, 0.25, 0.5, 1,2,4, 6,8, 24, 32, 48h after compound 11IV administration, 0.167, 0.5, 1,2,4, 6,8, 24, 32, 48h after PO administration, about 100 μ L of whole blood was collected at each time point, plasma was separated by centrifugation at 8000rpm for 6min, and plasma was frozen at-80 ℃ in a freezer. Plasma must be prepared within 30 minutes after blood collection.
Plasma sample analysis
The plasma sample adopts a liquid-liquid extraction method: mu.L of plasma was taken, 600. mu.L of MTBE (tert-butyl methyl ether) solution containing CH5424802 (10 ng/mL) was added, vortexed at 1500 rpm for 10min, then centrifuged at 12000 rpm for 5min, 300. mu.L of supernatant was taken, blown dry under nitrogen, redissolved with 300. mu.L of acetonitrile/water (7: 3, V/V), vortexed and mixed well, and analyzed by LC-MS/MS.
TABLE 3.1 rat PK evaluation results (iv:1mg/kg)
TABLE 3.2 rat PK evaluation results (po:2mg/kg)
AUClastArea under curve 0 → t when representing drug
CL stands for clearance
VssRepresenting steady state apparent distribution volume
T1/2Represents the half-life
TmaxRepresenting the peak time of blood
CmaxRepresenting blood drug peak concentration
F% represents absolute bioavailability
From the experimental results of tables 3.1 and 3.2, it is clear that compound 11 of the present invention has good pharmacokinetic properties.
Detailed Description
The present invention will be described in further detail with reference to the following examples. It should not be understood that the scope of the above-described subject matter of the present invention is limited to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
The following abbreviations represent the following definitions:
EA: ethyl acetate
PE: petroleum ether
DCM: methylene dichloride
NMP: n-methyl pyrrolidone
DMSO, DMSO: dimethyl sulfoxide
THF: tetrahydrofuran (THF)
TFA: trifluoroacetic acid
TBAF: tetrabutylammonium fluoride
DMF: dimethyl formamide
X-Phos: 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl
DDQ: 2, 3-dichloro-5, 6-dicyano-1, 4-benzoquinone
NBS: n-bromosuccinimide
Example 1: 7- (4- (3-oxa-8-azabicyclo [ 3.2.1)]Octane-8-yl) piperidin-1-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 1)
(1) Preparation of 2- (4-bromothien-2-yl) -1, 3-dioxolane
4-bromothiophene-2-carbaldehyde (19.1g,0.10mol) and methanesulfonic acid (1.0g,0.010mol) were dissolved in ethylene glycol (200mL), heated under reflux and water was allowed to separate for 6 hours. The resulting solution was rotary evaporated to remove the solvent, and the residue was isolated by silica gel column chromatography (petroleum ether: ethyl acetate =10:1) to give the product (16g, yield 68%).
1H-NMR(400MHz,CDCl3)δ:7.22(d,J=1.2,1H),7.08(d,J=0.8,1H),3.99～4.13(m,4H).
(2) Preparation of (5-Formylthiophen-3-yl) boronic acid
2- (4-bromothien-2-yl) -1, 3-dioxolane (16g,0.068mol) and triisopropyl borate (14.1g,0.075mol) were dissolved in dry THF (tetrahydrofuran) (150mL), n-BuLi (n-butyllithium) (30mL,2.5M) was slowly added at-78 ℃ under nitrogen protection, and after completion of the addition, stirring was carried out for 1 hour, the temperature was raised to room temperature, 10% hydrochloric acid was added, and stirring was carried out for 1 hour. Extraction with ethyl acetate, drying over anhydrous sodium sulfate and rotary evaporation to remove the solvent gave the product (7.5g, 71% yield).
1H-NMR(400MHz,CD3OD)δ:9.92(s,1H),8.33(s,1H),8.16(s,1H).
(3) Preparation of 1- (5-aldehyde thiophene-3-yl) hydrazine-1, 2-tert-butyl carboxylate
(5-Formylthiophen-3-yl) boronic acid (7.5g,0.0481mol) and azobistert-butyl ester (11.08g,0.0481mol) and a catalytic amount of copper acetate (0.44g,2.4mmol) were added to tetrahydrofuran (150mL) and stirred at room temperature for 16 hours. The solvent was removed by rotary evaporation, and the residue was subjected to silica gel column chromatography (petroleum ether: ethyl acetate =10:1) to isolate the product (11.7g, yield 71%).
(4) Preparation of 1- (5-cyanothiophen-3-yl) hydrazine-tert-butyl 1, 2-carboxylate
Tert-butyl 1- (5-formylthiophen-3-yl) hydrazine-1, 2-dicarboxylate (11.7g,0.0342mol) and hydroxylamine hydrochloride (4.8g,0.0691mol) were dissolved in pyridine (60mL), stirred at 90 ℃ for 10 minutes, cooled to room temperature, added with acetic anhydride (20.9g,0.205mol), heated to 80 ℃ and stirred for 1 hour, added with ethyl acetate (200mL), washed with a 10% hydrochloric acid solution, water, a saturated sodium chloride solution in that order, and the solvent was removed by rotary evaporation, and the product was isolated by silica gel column chromatography (ethyl acetate: petroleum ether =1:10) (10.4g, yield 90%).
(5) Preparation of 4-hydrazinothiophene-2-carbonitrile hydrochloride
Tert-butyl 1- (5-cyanothiophen-3-yl) hydrazine-1, 2-carboxylate (10.4g,0.031mol) was dissolved in dioxane (65mL), concentrated hydrochloric acid (39mL) was added, heated to 80 ℃ and stirred for 2 hours, and the solvent was removed by rotary evaporation to give the crude product (5.4 g).
(6) Preparation of 7-methoxy-5, 5-dimethyl-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
4-hydrazinothiophene-2-carbonitrile hydrochloride (5.4g,0.031mol) and 7-methoxy-1, 1-dimethyl-3, 4-dihydronaphthalen-2- (1H) -one (6.3g,0.031mol) were dissolved in trifluoroacetic acid (150mL), heated at reflux for 40 minutes, cooled, the reaction solution was poured into ethyl acetate and saturated potassium carbonate solution, the layers were separated, the organic phase was washed with saturated sodium chloride solution, dried, the solvent was removed by rotary evaporation, and the product was obtained by silica gel column separation (ethyl acetate: petroleum ether =1:5) (1.6g, yield 17%).
1H-NMR(400MHz,CDCl3)δ:8.32(s,1H),7.47(s,1H),7.24(m,1H),7.03(m,1H),6.84(m,1H),3.97(s,2H),3.86(s,3H),1.67(s,6H).
(7) Preparation of 7-methoxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
7-methoxy-5, 5-dimethyl-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (1.6g,5.2mmol) was dissolved in a mixed solvent of THF (tetrahydrofuran) (50mL) and water (5mL), cooled in an ice-water bath, DDQ (dichlorodicyanoquinone) (1.8g,7.9mmol) was added, then naturally warmed to room temperature and stirred for 30 minutes, ethyl acetate was extracted, the combined organic phases were washed with saturated potassium carbonate solution and water, dried, and the solvent was removed by rotary evaporation, and the residue was isolated by silica gel column chromatography (petroleum ether: ethyl acetate =1:1) to give the product (0.52g, 31% yield).
1H-NMR(400MHz,CDCl3)δ:8.91(s,1H),8.37(m,1H),7.54(s,1H),7.09(m,1H),7.03(m,1H),3.94(s,3H),1.78(s,6H).
(8) Preparation of 7-hydroxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
7-methoxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (0.52g,1.6mmol) and pyridine hydrochloride (2.96g,25.6mmol) were mixed well, microwave heated to 170 deg.C, stirred for 15 min, after which the mixture was taken up in water, extracted with ethyl acetate, the combined organics dried, and the solvent removed by rotary evaporation to give crude product (437 mg).
(9) Preparation of 2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate
Dissolving 7-hydroxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile crude product (437mg) in pyridine (10mL), slowly adding trifluoromethanesulfonic anhydride (440mg,1.56mmol) in ice-water bath, stirring at room temperature for 3 hours after the addition is finished, pouring the reaction solution into water, adding ethyl acetate for extraction, washing the combined organic phases with saturated saline, and removing the solvent by rotary evaporation to obtain crude product (462 mg).
1H-NMR(400MHz,CDCl3)δ:9.10(s,1H),8.51(d,J=8.8,1H),7.57(s,1H),7.53(d,J=2.4,1H),7.41(m,1H),1.83(s,6H).
(10) Preparation of 7- (4-hydroxypiperidin-1-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (440mg,1mmol) and 4-hydroxypiperidine (1.0g,0.01mol) were dissolved in N-methylpyrrolidone (30mL) and oil-bathed at 120 ℃ for 1 hour. After cooling, ethyl acetate (200mL) was added, the mixture was washed with saturated aqueous sodium chloride, and the solvent was removed by rotary evaporation to give the crude product (400 mg).
The molecular formula is as follows: c22H21N3O2S molecular weight: 391.49LC-MS (m/z): 392.0[ M + H]+
(11) Preparation of 5, 5-dimethyl-10-oxo-7- (4-oxopiperidin-1-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
7- (4-hydroxypiperidin-1-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thiophene [3,2-b ] indole-2-carbonitrile (400mg,1.02mmol) and dess-martin reagent (520mg,1.2mmol) were added to a mixed solution of dichloromethane (20mL) and dimethyl sulfoxide (8mL), and stirred at room temperature for 2 hours. Dichloromethane (200mL) was added, the mixture was washed with saturated sodium bicarbonate solution and saturated brine, the organic phase was dried, and the solvent was removed by rotary evaporation to give the crude product (230 mg).
The molecular formula is as follows: c22H19N3O2S molecular weight: 389.47LC-MS (m/z): 390.0[ M + H ]]+
(12) Preparation of 7- (4- (3-oxa-8-azabicyclo [3.2.1] octan-8-yl) piperidin-1-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
5, 5-dimethyl-10-oxo-7- (4-oxopiperidin-1-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (230mg,0.59mmol), indium chloride (130mg,0.59mmol) and 3-oxa-8-azabicyclo [3.2.1] octane hydrochloride (177mg,1.18mmol) were dissolved in N, N-dimethylformamide (15mL), and stirred at room temperature for 3 hours under nitrogen. Sodium cyanoborohydride (74mg,1.18mmol) was added, stirred at room temperature overnight, ethyl acetate (200mL) was added, washed with saturated aqueous sodium chloride solution, and subjected to preparative chromatography to give the final product (20mg, yield 7%).
The molecular formula is as follows: c28H30N4O2S molecular weight: 486.63LC-MS (m/z): 487.1[ M + H]+
1H-NMR(400MHz,MeOD)δ:8.13(d,J=8.8Hz,1H),7.74(s,1H),7.17-7.31(m,1H),7.08(m,1H),4.20(d,J=13.1Hz,2H),4.00(br.s.,2H),3.81-3.91(m,2H),3.67-3.79(m,2H),3.14(br.s.,1H),3.01(t,J=12.5Hz,2H),2.10～2.23(m,7H),1.77(s,6H),1.58-1.72(m,2H).
Example 27- (4- (8-oxa-3-azabicyclo [ 3.2.1)]Octane-3-yl) piperidin-1-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 2)
(1) Preparation of 2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate
7-hydroxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (10.1g,32.8mmol), prepared by the preparation method of step (1) to step (8) in example 1, was dissolved in anhydrous pyridine (50mL), trifluoromethanesulfonic anhydride (27.7g,98.2mmol) was added dropwise under ice bath, after completion of addition within 20min, and the reaction was continued at room temperature for 4H. Water (200mL) and ethyl acetate (200mL) were added to conduct liquid separation extraction, and after the organic phase was concentrated, the residue was purified by column chromatography (PE: EA =3:1) to obtain the product (7.9g, yield 55%).
(2) Preparation of 7- (4-hydroxypiperidin-1-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (400mg,0.91mmol) and 4-hydroxypiperidine (551mg,5.45mmol) were added to N-methylpyrrolidone (5mL), and the mixture was heated to 130 ℃ by microwave for 2 hours. After the reaction was completed, the reaction solution was poured into water (30mL), a solid was precipitated and suction-filtered under reduced pressure, and the resulting crude product was purified by column chromatography (DCM: MeOH =40:1) to obtain a product (200mg, yield 56%).
(3) Preparation of 5, 5-dimethyl-10-oxo-7- (4-oxopiperidin-1-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
7- (4-hydroxypiperidin-1-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (200mg,0.51mmol) was dissolved in DCM (10mL), DMSO (4mL) and triethylamine (4mL) were added, and sulfur trioxide pyridine complex (488mg,3.1mmol) was added in portions for 12H at room temperature. DCM and water were added to the system to extract and separate the liquid, the organic phase was concentrated and purified by column chromatography (DCM: MeOH =50:1) to give the product (150mg, yield 76%).
(4) Preparation of 7- (4- (8-oxa-3-azabicyclo [3.2.1] octan-3-yl) piperidin-1-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
5, 5-dimethyl-10-oxo-7- (4-oxopiperidin-1-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (150mg,0.39mmol) was dissolved in THF (5mL) and methanol (5mL), HOAC (0.5mL) and 8-oxa-3-azabicyclo [3.2.1] octane hydrochloride (105mg,0.7mmol) were added, respectively, and the reaction was continued at room temperature for 1H. Sodium cyanoborohydride (73mg,1.16mmol) was then added portionwise under ice bath, and the reaction was allowed to warm to room temperature for 4 h. Adding water to quench and react, rotary evaporating to remove organic solvent, adding EA and water to extract and separate liquid, concentrating the organic phase, and purifying by reverse phase preparative chromatography to obtain the product (40mg, 21% yield).
The molecular formula is as follows: c28H30N4O2S molecular weight: 486.63LC-MS (m/z): 487.3[ M + H]+
1H-NMR(400MHz,DMSO-d6)δ:12.72(s,1H),8.09(s,1H),7.97-7.94(d,1H,J=8.8Hz),7.17(s,1H),7.01-6.98(d,1H,J=8.8Hz),4.21(s,2H),3.95-3.91(m,2H),2.97-2.88(m,2H),2.61-2.56(m,2H),2.32-2.26(m,3H),1.89-1.53(m,12H),1.47-1.41(m,2H).
Example 37- (4- (3-oxa-8-azabicyclo [ 3.2.1)]Octane-8-yl) piperidin-1-yl) -8-ethyl-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 3)
(1) Preparation of 7-methoxy-1, 1-dimethyl-3, 4-dihydronaphthalen-2 (1H) -one
7-methoxy-3, 4-dihydronaphthalen-2 (1H) -one (50g,0.28mol), methyl iodide (100g, 0.70mol) were dissolved in THF (500mL), 50% aqueous KOH (125g, 1.1mol) was added, reflux was carried out for 2 hours, the reaction was stopped, ethyl acetate (2L) was added and extraction was carried out twice, drying was carried out, the solvent was removed by rotary evaporation, methanol (500mL) and water (250mL) were added, and the product was crystallized (45g, 79% yield).
(2) Preparation of 6-bromo-7-methoxy-1, 1-dimethyl-3, 4-dihydronaphthalen-2 (1H) -one
7-methoxy-1, 1-dimethyl-3, 4-dihydronaphthalen-2 (1H) -one (30g,0.15mol) was added to acetonitrile (600mL), and under stirring at room temperature, NBS (28.5g,0.16mol) was added thereto, stirred for 4 hours, added to 1L of water, stirred for 30 minutes, and dried by suction filtration to give the product (38g, 91% yield).
(3) Synthesis of 4-bromothiophene-2-carbonitrile
4-bromothiophene-2-carbaldehyde (600g,3.14mol) and hydroxylamine hydrochloride (438g,6.30mol) were added to pyridine (5L), heated to 90 ℃ for 10 minutes, then cooled to room temperature, added dropwise with acetic anhydride (1940g,19.0mol), heated to 80 ℃, reacted for 1 hour, poured into water (20L), stirred for 30 minutes, filtered with suction, and dried to obtain the product (564g, 95% yield).
(4) Synthesis of 4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) thiophene-2-carbonitrile
4-bromothiophene-2-carbonitrile (564g, 3.0mol), bis-pinacol boronate (805g,3.16mol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (56g), potassium acetate (885g, 9.02mol) dioxane (9L) were added to 2 four-necked bottles on average, heated to 90 ℃ under nitrogen protection, reacted for 15h, stopped, solvent removed by rotary evaporation, and the residue was purified by column chromatography (petroleum ether: ethyl ester =10:1) to give the product (426g, 60% yield).
(5) Preparation of di-tert-butyl 1- (5-cyanothiophen-3-yl) hydrazine-1, 2-dicarboxylate
4- (4,4,5, 5-tetramethyl-1, 3, 2-diheterocyclopentylboran-2-yl) thiophene-2-carbonitrile (426g,1.8mol), copper acetate (18g,0.09mol), di-tert-butyl (Z) -diazene-1, 2-dicarboxylate (621g,2.7mol) was added to THF (6L), stirred at room temperature for 16h, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography (petroleum ether: ethyl acetate =10:1) to give the product (396g, 65% yield).
(6) Preparation of 4-hydrazinothiophene-2-carbonitrile hydrochloride
Di-tert-butyl 1- (5-cyanothiophen-3-yl) hydrazine-1, 2-dicarboxylate (396g,1.17mol) was dissolved in dioxane (3L), concentrated hydrochloric acid (1584mL) was added and stirred at 80 ℃ for 2h, and the solvent was removed by rotary evaporation to give the product (140g, 68% yield).
(7) Preparation of 8-bromo-7-methoxy-5, 5-dimethyl-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
6-bromo-7-methoxy-1, 1-dimethyl-3, 4-dihydronaphthalen-2 (1H) -one (28.5g,0.1mol), 4-hydrazinothiophene-2-carbonitrile hydrochloride (17.6g,0.1mol) were dissolved in trifluoroacetic acid (200mL), heated under reflux for 40 minutes, cooled to room temperature, added with a saturated aqueous solution of ethyl acetate and sodium carbonate, separated, the organic phase washed with a saturated aqueous solution of sodium bicarbonate, dried, and the residue purified by column chromatography (petroleum ether: ethyl acetate =1:1) to give the product (17.6g, 45% yield).
(8) Synthesis of 8-bromo-7-methoxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
8-bromo-7-methoxy-5, 5-dimethyl-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (17.6g,0.045mol) was dissolved in THF (200mL), water (20mL) was added, DDQ (20.7g,0.09mol) was added in an ice-water bath, after completion of the addition, the temperature was raised to room temperature, stirring was carried out for 15 hours, the reaction was completed, the solvent was removed by rotary evaporation, and the residue was isolated and purified by column chromatography (petroleum ether: ethyl acetate =1:1) to give a product (9.5g, 52% yield).
(9) Synthesis of 8-bromo-7-hydroxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
8-bromo-7-methoxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (9g,0.02mol) and pyridine hydrochloride (45g) were mixed well, microwave reacted at 170 ℃ for 15 minutes, the mixture was dissolved in water, extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to give the product (5.6g, 72% yield).
(10) Preparation of 8-bromo-2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate
8-bromo-7-hydroxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (5.0g,12.9mmol) was dissolved in pyridine (100mL), trifluoromethanesulfonic anhydride (4.0g,14.2mmol) was slowly added under ice-bath, after completion of addition, stirring was performed at room temperature for 3 hours, ethyl acetate was added for extraction, and washed successively with water and saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to give the product (5.2g, 78% yield).
(11) Preparation of 8-bromo-7- (4-hydroxypiperidin-1-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
8-bromo-2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (1.0g,1.9mmol) and 4-hydroxypiperidine (1.92g,19mmol) were added to NMP (20mL), oil-washed at 120 ℃ for 5 hours, ethyl acetate (100mL) was added, washed successively with water, a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and the residue was subjected to silica gel column chromatography (PE: EA =1:1) to obtain a product (700mg, yield 77%).
(12) Preparation of 8-bromo-5, 5-dimethyl-10-oxo-7- (4-oxopiperidin-1-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Dissolving 8-bromo-7 (4-hydroxypiperidin-1-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (640mg,1.36mmol) in DCM (8mL) and DMSO (4mL), adding triethylamine (3mL), stirring at room temperature, adding pyridine trioxide (2.16g,13.7mmol), after completion of the addition, stirring at room temperature overnight, detecting by TLC, completely reacting, removing the solvent by rotary evaporation, adding water, stirring to precipitate, suction-filtering to obtain a filter cake, and purifying the filter cake by silica gel column chromatography (dichloromethane: methanol =30:1) to obtain a product (500mg, 78% yield).
(13) Preparation of 7- (4- (3-oxa-8-azabicyclo [3.2.1] octan-8-yl) piperidin-1-yl) -8-bromo-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
8-bromo-5, 5-dimethyl-10-oxo-7- (4-oxopiperidin-1-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (500mg,1.07mmol) and 3-oxa-8-azabicyclo [3.2.1] octane hydrochloride (319mg,213mmol) were dissolved in DMF (10mL), indium trichloride (237mg, 1.07mmol) was added, stirring was carried out at room temperature overnight, sodium cyanoborohydride (134mg,2.13mmol) was added, after stirring at room temperature for 5 hours, water (50mL) was added and the precipitate was stirred, suction filtered, and the filter cake was purified by column chromatography (dichloromethane: methanol =20:1) to give the product (510mg, 84% yield).
(14) Preparation of 7- (4- (3-oxa-8-azabicyclo [3.2.1] octan-8-yl) piperidin-1-yl) -5, 5-dimethyl-10-oxo-8- ((triisopropylsilyl) ethynyl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Adding 7- (4- (3-oxa-8-azabicyclo [3.2.1] octane-8-yl) piperidin-1-yl) -8-bromo-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (510mg,0.9mmol), triisopropylethynyl (246mg,1.35mmol), cesium carbonate (1.31g,4.02mmol), X-Phos (129mg,0.27mmol) and bis (acetonitrile) palladium dichloride (23mg,0.09mmol) to acetonitrile (20mL), reacting at 80 ℃ for 4 hours under nitrogen protection, detecting by TLC for completion of the reaction, cooling to room temperature, adding ethyl acetate (100mL) and saturated aqueous sodium chloride (50mL), separating to obtain an organic phase, the aqueous phase was extracted with ethyl acetate (50mL), the organic phases combined, dried over anhydrous sodium sulfate, and the solvent removed by rotary evaporation to give the crude product (650mg) which was used in the next step without purification.
(15) Preparation of 7- (4- (3-oxa-8-azabicyclo [3.2.1] oct-8-yl) piperidin-1-yl) -8-ethynyl-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Crude 7- (4- (3-oxa-8-azabicyclo [3.2.1] octan-8-yl) piperidin-1-yl) -5, 5-dimethyl-10-oxo-8- ((triisopropylsilyl) ethynyl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (650mg) was dissolved in THF (10mL), TBAF (380mg, 1.45mmol) was added, after stirring at room temperature for 1 hour, water and ethyl acetate were added each 50mL, and layers were separated to give an organic phase, which was washed six times with water, dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography (dichloromethane: methanol =20:1) to give a product (350 mg).
(16) Preparation of 7- (4- (3-oxa-8-azabicyclo [3.2.1] octan-8-yl) piperidin-1-yl) -8-ethyl-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Reacting 7- (4- (3-oxa-8-azabicyclo [3.2.1]]Octane-8-yl) piperidin-1-yl) -8-ethynyl-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Indole-2-carbonitrile (100mg,0.20mmol) was dissolved in THF (6mL) and methanol (4mL), Pd/C (60mg) was added, stirred at room temperature under hydrogen for 2 hours, filtered with suction, and the filtrate was purified by column chromatography (dichloromethane: methanol =15:1) by rotary evaporation to remove the solvent to give the final product (70mg, 69% yield). The molecular formula is as follows: c30H34N4O2S molecular weight: 514.69LC-MS (m/z): 515.3[ M + H]+
1H-NMR(400MHz,DMSO-d6)δ:12.81(s,1H),8.12(s,1H),7.98(s,1H),7.30(s,1H),3.51-3.54(m,2H),3.41-3.47(m,2H),3.15-3.19(m,2H),2.65-2.77(m,4H),2.11-2.28(m,1H),1.90-1.99(m,3H),1.66-1.75(m,11H),1.42-1.49(m,2H),1.18-1.29(m,3H).
Example 45, 5-dimethyl-7- (8-morpholin-3-azabicyclo [ 3.2.1)]Octane-3-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 4)
(1) Preparation of tert-butyl 8-morpholine-3-azabicyclo [3.2.1] octane-3-carboxylate
Tert-butyl 8-oxo-3-azabicyclo [3.2.1] octane-3-carboxylate (600mg,2.66mmol) was dissolved in THF (5mL) and methanol (5mL), and acetic acid (0.5mL) and morpholine (255mg,2.93mmol) were added, respectively, and the reaction was continued at room temperature for 1 hour. Sodium cyanoborohydride (336mg,5.3mmol) was then added portionwise in an ice bath and the reaction was continued at room temperature for 4 hours. Adding water to quench and react, performing rotary evaporation to remove the organic solvent, adding ethyl acetate and water to perform extraction and liquid separation, performing rotary evaporation to remove the solvent from the obtained organic phase, and purifying the obtained crude product by column chromatography (PE: EA =10:1) to obtain a product (408mg, yield 52%). .
(2) Preparation of 4- (3-azabicyclo [3.2.1] octan-8-yl) morpholine
Tert-butyl 8-morpholine-3-azabicyclo [3.2.1]]Octane-3-carboxylate (400mg,1.35mmol) was dissolved in DCM (20mL), and TFA (10mL) was added, and the reaction was continued at room temperature for 2 hours. The system is rotary evaporated to remove the solvent and then saturated aqueous sodium bicarbonate solution is added to adjust the pH to 9, the solvent is removed by rotary evaporation and the residue is chromatographed (DCM: MeOH: Et)3N =40:1:0.1) to give the product (215mg, yield 81%).
(3) Preparation of 5, 5-dimethyl-7- (8-morpholine-3-azabicyclo [3.2.1] octan-3-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (161mg,0.37mmol), 4- (3-azabicyclo [3.2.1] oct-8-yl) morpholine (215mg,1.1mmol) prepared according to the preparation method of step (1) in example 2 was dissolved in N-methylpyrrolidone (10mL), N-diisopropylethylamine (472mg,3.7mmol) was added thereto, and the reaction was continued at 120 ℃ for 2 hours under microwave. After completion of the reaction, the reaction mixture was poured into water (20mL), and the precipitated solid was suction-filtered under reduced pressure and the obtained solid was purified by column chromatography (DCM: MeOH =20:1) to obtain the final product (35mg, yield 20%).
The molecular formula is as follows: c28H30N4O2S molecular weight: 486.63LC-MS (m/z): 487.3[ M + H]+
1H-NMR(400MHz,CDCl3)δ:9.07(s,1H),8.27(d,1H,J=8.8Hz),7.52(s,1H),6.89-6.95(m,2H),3.71-3.73(m,4H),3.41-3.43(m,2H),3.33-3.35(m,2H),2.53-2.57(m,4H),2.44(s,2H),2.28-2.30(m,1H),1.74-1.87(m,10H).
Example 55, 5-dimethyl-7- (3-morpholin-8-azabicyclo [ 3.2.1)]Octane-8-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 5)
(1) Preparation of 7- (3-hydroxy-8-azabicyclo [3.2.1] octan-8-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (400mg,0.91mmol) prepared in the preparation method of the step (1) in example 2 and 8-azabicyclo [3.2.1] octan-3-ol (693mg,5.45mmol) were added to N-methylpyrrolidone (5mL) and reacted at 130 ℃ for 2 hours under microwave. After the reaction, the system was poured into 30mL of water, and a solid precipitated out of the system and was filtered under reduced pressure, and the resulting crude product was purified by column chromatography (DCM: MeOH =40:1) to give the product (200mg, 53% yield).
(2) Preparation of 5, 5-dimethyl-10-oxo-7- (3-oxo-8-azabicyclo [3.2.1] octan-8-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
7- (3-hydroxy-8-azabicyclo [3.2.1] octan-8-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (200mg,0.48mmol) was dissolved in 10mL DCM, DMSO (4mL) and triethylamine (4mL) were added, and pyridine complex (458mg,2.9mmol) was added in portions and reacted at room temperature for 12 hours. DCM was added to the system and water was extracted for liquid separation, the resulting organic phase was evaporated with rotary evaporation to remove the solvent and purified by column chromatography (DCM: MeOH =50:1) to give the product (159mg, 80% yield).
(3) Preparation of 5, 5-dimethyl-7- (3-morpholine-8-azabicyclo [3.2.1] octan-8-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
5, 5-dimethyl-10-oxo-7- (3-oxo-8-azabicyclo [3.2.1] octan-8-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (159mg,0.38mmol) was dissolved in THF (5mL) and methanol (5mL), and acetic acid (0.5mL) and morpholine (67mg,0.77mmol) were added, respectively, and the reaction was continued at room temperature for 1 hour. Sodium cyanoborohydride (73mg,1.16mmol) was then added portionwise under ice bath and the reaction was continued back to room temperature for 4 hours. Adding water to quench and react, performing rotary evaporation to remove organic solvent, adding EA and water to perform extraction and liquid separation, performing rotary evaporation to remove solvent from the obtained organic phase, and purifying by reverse phase preparative chromatography to obtain the final product (40mg, yield 22%).
The molecular formula is as follows: c28H30N4O2S molecular weight: 486.63LC-MS (m/z): 487.3[ M + H]+
1H-NMR(400MHz,DMSO-d6)δ:12.71(s,1H),8.08(s,1H),7.97-7.94(d,1H,J=8.4Hz),7.04(s,1H),6.89-6.87(m,1H),4.53(s,2H),3.44-3.49(m,4H),2.29(s,4H),1.99-1.97(m,3H),1.87-1.67(m,2H),1.78-1.41(m,8H),1.55-1.40(m,2H).
Example 65, 5-dimethyl-7- (3-methyl-3, 8-diazabicyclo [ 3.2.1)]Octane-8-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 6)
(1) Preparation of tert-butyl 8- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -3, 8-diazabicyclo [3.2.1] octane-3-carboxylate
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (250mg,0.57mmol) prepared in the preparation method of the step (1) in example 2 and tert-butyl 3, 8-diazabicyclo [3.2.1] octane-3-carboxylate (483mg,2.28mmol) were added to N-methylpyrrolidone (20mL) and reacted at 120 ℃ for 24 hours in an oil bath. After completion of the reaction, the reaction mixture was poured into water (30mL), a solid was precipitated and suction-filtered under reduced pressure, and the crude product was purified by column chromatography (DCM: MeOH =40:1) to obtain a product (80mg, yield 28%).
(2) Preparation of 7- (3, 8-diazabicyclo [3.2.1] octan-8-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Tert-butyl 8- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -3, 8-diazabicyclo [3.2.1] octane-3-carboxylate (80mg,0.16mmol) was dissolved in DCM (10mL), and TFA (5mL) was added and the reaction was continued at room temperature for 2 hours. The reaction solution was then rotary evaporated to remove the solvent, saturated aqueous sodium bicarbonate was added to adjust the pH to 9, DCM and water were added for extraction and liquid separation, and the organic phase was rotary evaporated to remove the solvent to give the product (60mg, 93% yield).
(3) Preparation of 5, 5-dimethyl-7- (3-methyl-3, 8-diazabicyclo [3.2.1] octan-8-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
7- (3, 8-diazabicyclo [3.2.1] octan-8-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (60mg,0.15mmol) was dissolved in acetonitrile (10mL), followed by addition of aqueous formaldehyde (61mg,0.75mmol), and the reaction was continued at room temperature for 1 hour. Sodium cyanoborohydride (19mg,0.3mmol) was then added portionwise under ice bath and the reaction was continued back to room temperature for 4 hours. Adding water to quench reaction, rotary evaporating to remove organic solvent, adding EA and water to extract and separate, rotary evaporating the obtained organic phase to remove solvent, and purifying the residue by reversed phase preparative chromatography to obtain the final product (12mg, 19% yield).
The molecular formula is as follows: c24H24N4Molecular weight of OS: 416.54LC-MS (m/z): 417.2[ M + H]+
1H-NMR(400MHz,CDCl3)δ:10.94(s,1H),8.26-8.23(d,1H,J=8.8Hz),7.48(s,1H),6.88(s,1H),6.84-6.82(d,1H,J=9.2Hz),4.37(s,2H),2.66-2.64(m,2H),2.46-2.44(m,2H),2.17(s,3H),2.11-2.04(m,4H),1.87(s,6H).
Example 75, 5-dimethyl-7- (8-methyl-3, 8-diazabicyclo [3.2.1]Octane-3-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 7)
(1) Preparation of 8-benzyl-3-tert-butyl-3, 8-diazabicyclo [3.2.1] octane-3, 8-dicarboxylate
Tert-butyl 3, 8-diazabicyclo [3.2.1] octane-3-carboxylate (820mg,3.86mmol), N-diisopropylethylamine (1.0g,7.7mmol) and dichloromethane (50mL) were added dropwise over ice, benzyl chloroformate (990mg,5.8mmol) was added dropwise over 20 minutes, and the reaction was allowed to warm to room temperature for 4 hours. The reaction was quenched with water, extracted with ethyl acetate, and the solvent removed by rotary evaporation of the organic phase to give the product (1.09g, 81% yield).
(2) Preparation of benzyl 3, 8-diazabicyclo [3.2.1] octane-8-carboxylate
8-benzyl 3-tert-butyl 3, 8-diazabicyclo [3.2.1] octane-3, 8-dicarboxylate (1.09g,3.15mmol) was dissolved in dichloromethane (20mL), followed by addition of TFA (10mL), and the reaction was continued at room temperature for 2 hours. The system was then rotary evaporated to remove the solvent, saturated aqueous sodium bicarbonate was added to adjust the pH to 9, dichloromethane and water were added for extraction and separation, the organic phase was rotary evaporated to remove the solvent and the crude product was purified by reverse phase preparative chromatography to give the product (715mg, 92% yield).
(3) Preparation of benzyl 3- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylate
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (426mg,0.97mmol), benzyl 3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (715mg,2.9mmol) prepared according to the preparation method of step (1) in example 9 was dissolved in N-methylpyrrolidone (10mL), and N, N-diisopropylethylamine (1.25g,9.7mmol) was added thereto, and the reaction was continued at 120 ℃ for 2 hours under microwave. After completion of the reaction, the reaction mixture was poured into water (20mL), and the precipitated solid was suction-filtered under reduced pressure and purified by column chromatography (DCM: MeOH =50:1) to give the product (429mg, yield 82%).
(4) Preparation of 7- (3, 8-diazabicyclo [3.2.1] octan-3-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Benzyl 3- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (429mg,0.80mmol) was added to acetonitrile (10mL), and iodotrimethylsilane (0.8g,4mmol) was slowly added dropwise at room temperature, and the reaction was continued for 2 hours after completion of the addition. The reaction was quenched by the addition of water (1mL) to the total, the organic solvent was removed by rotary evaporation, and the residue was purified by column chromatography (DCM: MeOH =40:1) to give the product (200mg, 62% yield).
(5) Preparation of 5, 5-dimethyl-7- (8-methyl-3, 8-diazabicyclo [3.2.1] octan-3-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
7- (3, 8-diazabicyclo [3.2.1] octan-3-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (200mg,0.50mmol) was dissolved in acetonitrile (10mL), and 37% aqueous formaldehyde (202mg,2.5mmol) was added to continue the reaction at room temperature for 1 hour. Sodium cyanoborohydride (63mg,1mmol) was then added portionwise under ice bath and the reaction was continued back to room temperature for 4 h. Water quenching was added to the reaction mixture, the organic solvent was removed by rotary evaporation, ethyl acetate and water were added for extraction and liquid separation, the solvent was removed from the organic phase by rotary evaporation, and the residue was purified by column chromatography (DCM: MeOH =20:1) to give the final product (80mg, 39% yield).
The molecular formula is as follows: c24H24N4Molecular weight of OS: 416.54LC-MS (m/z): 417.2[ M + H]+
1H-NMR(400MHz,MeOD)δ:8.09(d,1H,J=8.8Hz,),7.71-7.73(m,1H),7.08(d,1H,J=2.0Hz),6.94-6.97(m,1H),3.76-3.79(m,2H),3.61(s,2H),3.17-3.20(m,2H),2.56(s,3H),2.18-2.20(m,2H),1.90-1.93(m,2H),1.75(s,6H).
Example 85, 5-dimethyl-7- (7-methyl-2, 7-diazaspiro [3.5]]Nonan-2-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 8)
(1) Preparation of tert-butyl 2- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (100mg,0.23mmol) prepared according to the preparation method of step (1) in example 13 and tert-butyl 2, 7-diazaspiro [3.5] nonane-7-carboxylate (520mg,2.3mmol) were added to NMP (5mL), oil-washed at 120 ℃ for 3 hours, ethyl acetate (50mL) was added, washed successively with water and a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (PE: = EA 1:1) to obtain a product (100mg, yield 85%).
(2) Preparation of 5, 5-dimethyl-10-oxo-7- (2, 7-diazaspiro [3.5] nonan-2-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Tert-butyl 2- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (100mg,0.19mmol) was dissolved in DCM (4mL), trifluoroacetic acid (4mL) was added, the solvent was removed by rotary evaporation after stirring at room temperature for 0.5 hour, aqueous ammonia and methanol were added, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (dichloromethane: methanol =15:1) to give a product (70mg, yield 89%).
(3) Preparation of 5, 5-dimethyl-7- (7-methyl-2, 7-diazaspiro [3.5] nonan-2-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
5, 5-dimethyl-10-oxo-7- (2, 7-diazaspiro [3.5] nonan-2-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (70mg,0.17mmol) was dissolved in a mixed solvent of methanol (3mL), tetrahydrofuran (3mL) and acetic acid (0.5mL), formaldehyde (73mg) was added, after stirring at room temperature for 1 hour, sodium cyanoborohydride (53mg,0.84mmol) was added, stirring at room temperature overnight, water (50mL) was added, the precipitate was stirred, filtered, and the filter cake was washed with methanol to give the final product (40mg, 55% yield).
The molecular formula is as follows: c25H26N4Molecular weight of OS: 430.57LC-MS (m/z): 431.2[ M + H]+
1H-NMR(400MHz,DMSO-d6)δ:12.64(s,1H),8.05(s,1H),7.96(d,1H,J=8.8Hz),6.61(s,1H),6.45-6.48(m,1H),3.68(s,4H),2.26-2.49(m,4H),2.13(s,3H),1.75(s,4H),1.61-1.68(m,6H).
Example 95, 5-dimethyl-7- (2-methyl-2, 7-diazaspiro [3.5]]Nonan-7-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 9)
(1) Preparation of 2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate
7-hydroxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (5.0g,16.2mmol) was dissolved in pyridine (100mL), trifluoromethanesulfonic anhydride (5.02g, 17.9mmol) was slowly added under ice bath, after completion of addition, stirring at room temperature for 3 hours, extraction was performed by adding ethyl acetate, and then washing with water and saturated aqueous sodium chloride solution in this order, drying over anhydrous sodium sulfate, and spin-drying to give the title compound (5.0g, yield 70%).
(2) Preparation of tert-butyl 7- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -2, 7-diazaspiro [3.5] nonane-2-carboxylate
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (100mg,0.23mmol) prepared in the preparation method of the step (1) in example 12 and tert-butyl 2, 7-diazaspiro [3.5] nonane-2-carboxylate (520mg,2.3mmol) were added to 5mL of NMP, oil-bathed at 120 ℃ for 5 hours, 50mL of ethyl acetate was added, washed successively with water, a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (PE: EA =1:1) to obtain a product (110mg, yield 93%).
(3) Preparation of 5, 5-dimethyl-10-oxo-7- (2, 7-diazaspiro [3.5] nonan-7-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Tert-butyl 7- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -2, 7-diazaspiro [3.5] nonane-2-carboxylate (100mg,0.19mmol) was dissolved in DCM (4mL), trifluoroacetic acid (4mL) was added, the mixture was stirred at room temperature for 0.5 hour, then the solvent was removed by rotary evaporation, aqueous ammonia and methanol were added, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (dichloromethane: methanol =15:1) to give a product (79mg, yield 98%).
(4) Preparation of 5, 5-dimethyl-7- (2-methyl-2, 7-diazaspiro [3.5] nonan-7-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
5, 5-dimethyl-10-oxo-7- (2, 7-diazaspiro [3.5] nonan-7-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (70mg,0.17mmol) was dissolved in a mixed solvent of methanol (3mL), tetrahydrofuran (3mL) and acetic acid (0.5mL), formaldehyde (37% aqueous solution) (73mg,0.9mmol) was added, after stirring at room temperature for 1 hour, sodium cyanoborohydride (53mg,0.84mmol) was added, stirring at room temperature overnight, water (50mL) was added, the precipitate was stirred, filtered, and the filter cake was washed with methanol to give the final product (53mg, 73% yield).
The molecular formula is as follows: c25H26N4Molecular weight of OS: 430.57LC-MS (m/z): 431.2[ M + H]+
1H-NMR(400MHz,DMSO-d6)δ:12.80(s,1H),8.09(s,1H),7.96(d,1H,J=8.8Hz),7.19(d,1H,J=2Hz),7.02-7.05(m,1H),3.62-3.78(m,4H),3.37-3.38(m,4H),2.70(s,3H),1.86(s,4H),1.71(s,6H).
Example 1055-dimethyl-7- (6-methyl-2, 6-diazaspiro [3.3]]Heptane-2-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 10)
(1) Preparation of tert-butyl 6- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -2, 6-diazaspiro [3.3] heptane-2-carboxylate
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (100mg,0.23mmol) prepared according to the preparation method of step (1) of example 9 and tert-butyl 2, 6-diazaspiro [3.3] heptane-2-carboxylate (455mg,2.3mmol) were added to NMP (5mL), oil-bathed at 120 ℃ for 3 hours, ethyl acetate (50mL) was added, washed successively with water, a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (PE: EA =1:1) to obtain a product (100mg, yield 90%).
(3) Preparation of 5, 5-dimethyl-10-oxo-7- (2, 6-diazaspiro [3.3] heptan-2-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Tert-butyl 6- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -2, 6-diazaspiro [3.3] heptane-2-carboxylate (100mg,0.20mmol) was dissolved in DCM (4mL), trifluoroacetic acid (4mL) was added, the solvent was removed by rotary evaporation after stirring at room temperature for 0.5 hour, aqueous ammonia and methanol were added, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (dichloromethane: methanol =15:1) to give a product (70mg, yield 88%).
(4) Preparation of 5, 5-dimethyl-7- (6-methyl-2, 6-diazaspiro [3.3] heptan-2-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
5, 5-dimethyl-10-oxo-7- (2, 6-diazaspiro [3.3] heptan-2-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (66mg,0.17mmol) was dissolved in a mixed solvent of methanol (3mL), tetrahydrofuran (3mL) and acetic acid (0.5mL), formaldehyde (37% aqueous solution) (73mg,0.9mmol) was added, after stirring for 1 hour at room temperature, sodium cyanoborohydride (53mg,0.84mmol) was added, stirring overnight at room temperature, 50mL of water was added, the precipitate was stirred, filtered, and the filter cake was washed with methanol to give the final product (30mg, 44% yield).
The molecular formula is as follows: c23H22N4Molecular weight of OS: 402.52LC-MS (m/z): 403.2[ M + H+]
1H-NMR(400MHz,DMSO-d6)δ:12.80(s,1H),8.08(s,1H),7.97(d,J=8.4Hz,1H),6.63(s,1H),6.48-6.50(m,1H),4.09(s,4H),3.76(s,4H),2.49(s,3H),1.69(s,6H).
Example 115, 5-dimethyl-7- (9-methyl-3, 9-diazaspiro [5.5]]Undec-3-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 11)
(1) Preparation of tert-butyl 9- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -3, 9-diazaspiro [5.5] undecane-3-carboxylate
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (100mg,0.23mmol) prepared according to the preparation method of the first step (1) of example 9 and tert-butyl 3, 9-diazaspiro [5.5] undecane-3-carboxylate (585mg,2.3mmol) were added to NMP (5mL), oil-washed at 120 ℃ for 3 hours, ethyl acetate (50mL) was added, washed with water and a saturated aqueous sodium chloride solution in this order, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (PE: EA =1:1) to obtain a product (100mg, yield 78%).
(2) Preparation of 5, 5-dimethyl-10-oxo-7- (3, 9-diazaspiro [5.5] undecan-3-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Tert-butyl 9- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -3, 9-diazaspiro [5.5] undecane-3-carboxylate (100mg,0.18mmol) was dissolved in DCM (4mL), trifluoroacetic acid (4mL) was added, the mixture was stirred at room temperature for 0.5 hour, then the solvent was removed by rotary evaporation, aqueous ammonia and methanol were added, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (dichloromethane: methanol =15:1) to give a product (80mg, yield 98%).
(3) Preparation of 5, 5-dimethyl-7- (9-methyl-3, 9-diazaspiro [5.5] undecan-3-yl) -10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
5, 5-dimethyl-10-oxo-7- (3, 9-diazaspiro [5.5] undecan-3-yl) -5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (75mg,0.17mmol) was dissolved in a mixed solvent of methanol (3mL), tetrahydrofuran (3mL) and acetic acid (0.5mL), formaldehyde (73mg,0.85mmol) was added, after stirring at room temperature for 1 hour, sodium cyanoborohydride (53mg,0.84mmol) was added, stirring at room temperature overnight, water (50mL) was added, the precipitate was stirred, filtered, and the filter cake was washed with methanol to give the final product (50mg, 64% yield).
The molecular formula is as follows: c27H30N4Molecular weight of OS: 458.62LC-MS (m/z): 459.3[ M + H]+
1H-NMR(400MHz,CDCl3)δ:8.22-8.29(m,1H),7.49(s,1H),6.96-6.97(m,2H),3.37-3.40(m,4H),2.34-2.41(m,4H),2.27-2.32(m,3H),1.65-1.79(m,6H),1.52-1.61(m,4H),1.49-1.50(m,4H).
Example 125, 5-dimethyl-7- (cis-5-Methylpiperidino [3, 4-c)]Pyrrol-2- (1H) -yl) -10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 13)
(1) Preparation of 2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate
7-hydroxy-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (5.0g,16.2mmol) prepared according to the preparation method of example 1, step (1) to step (8) was dissolved in pyridine (100mL), trifluoromethanesulfonic anhydride (5.02g,17.8mmol) was slowly added under ice bath, after completion of addition, stirring at room temperature for 3 hours, extraction was performed by adding ethyl acetate, and washing was successively with water and a saturated aqueous sodium chloride solution, drying was performed over anhydrous sodium sulfate, and the solvent was removed by rotary evaporation to obtain the product (5.2g, yield 73%).
(2) Preparation of tert-butyl cis-5- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) hexahydropyrrolo [3,4-c ] pyrrole-2 (1H) -carboxylate
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (100mg,0.23mmol) and tert-butyl cis-hexahydropyrrolo [3,4-c ] pyrrole-2 (1H) -carboxylate (488mg,2.3mmol) were added to NMP (5mL), oil-bathed at 120 ℃ for 3 hours, ethyl acetate (50mL) was added, washed successively with water, saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (PE: EA =1:1) to obtain a product (100mg, yield 87%).
(3) Preparation of 7- (cis-hexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Tert-butyl cis-5- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) hexahydropyrrolo [3,4-c ] pyrrole-2 (1H) -carboxylate (100mg,0.20mmol) was dissolved in DCM (4mL), trifluoroacetic acid (4mL) was added, the mixture was stirred at room temperature for 0.5 hour, then the solvent was removed by rotary evaporation, aqueous ammonia and methanol were added, the solvent was removed by rotary evaporation, and the residue was purified by silica gel column chromatography (dichloromethane: methanol =15:1) to obtain a product (78mg, yield 97%).
(4) Preparation of 5, 5-dimethyl-7- (cis-5-methylhexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) -10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
7 (cis-hexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile (68mg,0.17mmol) was dissolved in a mixed solvent of methanol (3mL), tetrahydrofuran (3mL) and acetic acid (0.5mL), formaldehyde (73mg,0.89mmol) was added, after stirring at room temperature for 1 hour, sodium cyanoborohydride (53mg,0.84mmol) was added, stirring at room temperature overnight, water (50mL) was added, the precipitate was stirred, filtered, and the filter cake was washed with methanol to give the final product (40mg, 57% yield).
The molecular formula is as follows: c24H24N4Molecular weight of OS: 416.54LC-MS (m/z): 417.2[ M + H]+
1H-NMR(400MHz,DMSO-d6)δ:12.82(s,1H),8.10(s,1H),8.00(d,1H,J=8.8Hz),6.87(s,1H),6.77(d,1H,J=8.4Hz),3.35-3.85(m,9H),2.84(s,4H),1.71(s,6H).
Example 137- (Exo-6-amino-3-azabicyclo [ 3.1.0)]Hexane-3-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f]Thieno [3,2-b]Preparation of indole-2-carbonitrile (Compound 15)
(1) Preparation of tert-butyl exo-6- (((benzyloxy) carbonyl) amino) -3-azabicyclo [3.1.0] hexane-3-carboxylate
Tert-butyl exo-6-amino-3-azabicyclo [3.1.0] hexane-3-carboxylate (4.15g,21mmol), N, N-diisopropylethylamine (5.4g,42mmol) was added to dichloromethane (100mL), benzyl chloroformate (5.36g,31mmol) was added dropwise over ice, and after dropwise addition over 20 minutes, the reaction was allowed to warm to room temperature for 4 hours. Water (100mL) was added for extractive separation, and the solvent was removed by rotary evaporation of the organic phase to give the product (5.0g, 72% yield).
(2) Preparation of benzyl (exo-3-azabicyclo [3.1.0] hex-6-yl) carbamate
Tert-butyl exo-6- (((benzyloxy) carbonyl) amino) -3-azabicyclo [3.1.0] hexane-3-carboxylate (5.0g,15.0mmol) was dissolved in dichloromethane (20mL), and TFA (10mL) was added and the reaction was continued at room temperature for 2 hours. The solvent was then removed by rotary evaporation, saturated aqueous sodium bicarbonate was added to adjust the pH to 9, DCM was added and water was added for extraction and separation, the organic phase was rotary evaporated to remove the solvent and the crude product was purified by reverse phase preparative chromatography to give the product (3.0g, 86% yield).
(3) Preparation of benzyl (exo-3- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -3-azabicyclo [3.1.0] hex-6-yl) carbamate
2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl trifluoromethanesulfonate (360mg,0.82mmol), benzyl (exo-3-azabicyclo [3.1.0] hex-6-yl) carbamate (569mg,2.45mmol) prepared according to the preparation method of step (1) of example 2 were dissolved in N-methylpyrrolidone (10mL), N-diisopropylethylamine (1.06g,8.2mmol) was added, and the reaction was continued at 120 ℃ for 2 hours under microwave. After the reaction, the reaction mixture was poured into water, and the precipitated solid was vacuum filtered, and the obtained solid was purified by column chromatography (DCM: MeOH =50:1) to obtain a product (134mg, yield 31%).
(4) Preparation of 7- (exo-6-amino-3-azabicyclo [3.1.0] hex-3-yl) -5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indole-2-carbonitrile
Benzyl (exo-3- (2-cyano-5, 5-dimethyl-10-oxo-5, 10-dihydro-4H-benzo [ f ] thieno [3,2-b ] indol-7-yl) -3-azabicyclo [3.1.0] hex-6-yl) carbamate (134mg,0.26mmol) was added to acetonitrile (10mL), iodotrimethylsilane (260mg,1.3mmol) was slowly added dropwise at room temperature, and the reaction was continued for 2 hours after the addition. The reaction was quenched by addition of water, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography (DCM: MeOH =40:1) to give the final product (40mg, 40%).
The molecular formula is as follows: c22H20N4Molecular weight of OS: 388.49LC-MS (m/z): 389.2[ M + H]+
1H-NMR(400MHz,MeOD)δ:8.10(d,1H,J=8.8Hz),7.73(s,1H),6.80-6.81(m,1H),6.71-6.73(m,1H),3.86(d,2H,J=10Hz),3.51(d,2H,J=9.6Hz),2.54(s,1H),2.2(s,2H),1.76(s,6H)。

Claims (11)

1. A compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof:
wherein,
A1is selected from C-R1Or N;
A2is selected from C-R2Or N;
A3is selected from C-R3Or N, and A1、A2And A3Not simultaneously selected from N;
R1and R3Each independently selected from hydrogen, hydroxyl, carboxyl, amino, nitro, sulfonyl, halogen atom, cyano, C1-6Alkyl radical, C1-6Alkoxy group, (C)1-6Alkyl radical)2Amino group, C2-6Alkenyl radical, C2-6Alkynyl or 3-14 membered cycloalkyl;
R2selected from hydrogen, cyano, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylthio, 3-to 14-membered heterocyclyloxy, C2-6Alkenyl radical, C2-6Alkynyl, 3-14 membered cycloalkyl, 3-14 membered heterocyclyl, 5-15 membered heteroaryl or 6-14 membered aryl;
q is selected from 3-8 membered heterocyclic group, 6-14 membered heterocyclic group, 6-12 membered bridged ring group or 6-12 membered spiro ring group;
R4selected from the following groups:
(1) hydrogen, amino, hydroxy, nitro, cyano, halogen atom, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylamino radical, (C)1-6Alkyl radical)2Amino group, C1-6Alkylaminocarbonyl, hydroxy C1-6Alkyl, hydroxy C1-6Alkylamino, halogeno C1-6Alkyl radical, C1-6Alkylsulfonyl radical, C1-6Alkylsulfonylamino, aminosulfonyl, aminosulfonylamino, C2-6Alkenyl or C2-6An alkynyl group,
(2) a 6-12 membered bridged ring group, a 6-12 membered spiro ring group, a 3-8 membered heterocyclic group or a 6-14 membered heterocyclic group optionally substituted with a substituent selected from the group consisting of amino group, hydroxyl group, nitro group, halogen atom, carboxyl group, C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl radical, C2-6Alkynyl, 3-to 8-membered heterocyclic group or 3-to 8-membered cycloalkyl;
n is selected from 0, 1,2,3,4 or 5, and when n is 0, R is4When n is not less than 2, R4Which may be the same or different from each other,
and when Q is selected from 3-8 membered heterocyclic group, R4Can not be selected from 3-8 membered heterocyclic groups;
R5and R6Each independently selected from hydrogen, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy, hydroxy C1-6Alkyl radical, C2-6Alkenyl or C2-6An alkynyl group,
or R5And R6Are linked to each other to form a 3-to 14-membered heterocyclic group or a 3-to 14-membered cycloalkyl group together with the carbon atom to which they are linked;
R7selected from hydrogen, cyano, nitro, hydroxy, amino, amidino, sulfonyl, C1-6Alkyl radical, C1-6Alkoxy group, halogen atom, 3-to 14-membered cycloalkyl group, C2-6Alkenyl or C2-6Alkynyl, said C1-6Alkyl radical, C1-6Alkoxy, 3-to 14-membered cycloalkyl, C2-6Alkenyl and C2-6Alkynyl groups may be independently optionally substituted with the following substituents: hydroxyl, carboxyl, amino, cyano, halogen atom, nitro or 3-14-membered heterocyclic group;
m is selected from O, S or N-R8，R8Selected from hydrogen, C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl or C2-6Alkynyl, said C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl and C2-6Alkynyl may optionally be substituted by C1-6Alkoxy substitution;
y is selected from N or C-R9；
X is selected from O, S or N-R9；
R9Selected from hydrogen, hydroxyl, carboxyl, amino, nitro, cyano, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy group, (C)1-6Alkyl radical)2Amino group, C2-6Alkenyl radical, C2-6Alkynyl or 3-14 membered cycloalkyl.
2. A compound according to claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof, wherein formula (i) is formula (ii):
wherein,
R1and R3Each independently selected from hydrogen, hydroxyl, carboxyl, amino, nitro, sulfonyl, halogen atom, cyano, C1-6Alkyl radical, C1-6Alkoxy group, (C)1-6Alkyl radical)2Amino group, C2-6Alkenyl radical, C2-6Alkynyl or 3-to 8-membered cycloalkyl;
R2selected from hydrogen, cyano, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylthio, 3-to 14-membered heterocyclyloxy, C2-6Alkenyl radical, C2-6Alkynyl, 3-to 8-membered cycloalkyl, 5-to 6-membered heterocyclyl or 5-to 6-membered heteroaryl;
q is selected from 4-7 membered heterocyclic group, 6-12 membered heterocyclic group, 7-10 membered bridged ring group or 6-12 membered spiro ring group;
R4selected from the following groups:
(1) hydrogen, amino, hydroxy, nitro, cyano, halogen atom, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylamino radical, (C)1-4Alkyl radical)2Amino group, C1-4Alkylaminocarbonyl, hydroxy C1-4Alkyl, hydroxy C1-4Alkylamino, halogeno C1-4Alkyl radical, C1-4Alkylsulfonyl radical, C1-4Alkylsulfonylamino, aminosulfonyl, aminosulfonylamino, C2-6Alkenyl or C2-6An alkynyl group,
(2) 6-10 membered bridged ring group, 6-12 membered spiro ring group, 4-7 membered heterocyclic group or 6-12 membered heterocyclic ring optionally substituted with a substituent selected from amino group, hydroxyl group, halogen atom, C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl or C2-6An alkynyl group;
n is selected from 0, 1,2 or 3, and when n is 0, R4When n is not less than 2, R4Which may be the same or different from each other,
and when Q is selected from 4-7 membered heterocyclic group, R4Cannot be selected from 4-7 membered heterocyclic groups;
R5and R6Each independently selected from hydrogen, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy, hydroxy C1-6Alkyl radical, C2-6Alkenyl or C2-6An alkynyl group,
or R5And R6Are connected with each other to form a 5-to 10-membered heterocyclic group or a 3-to 8-membered cycloalkyl group together with the carbon atom to which they are connected;
R7selected from hydrogen, cyano, nitro, hydroxy, amino, amidino, sulfonyl, C1-6Alkyl radical, C1-6Alkoxy group, halogen atom, 3-to 8-membered cycloalkyl group, C2-6Alkenyl or C2-6Alkynyl, said C1-6Alkyl radical, C1-6Alkoxy, 3-to 8-membered cycloalkyl, C2-6Alkenyl and C2-6Alkynyl groups may be independently optionally substituted with the following substituents: hydroxyl, carboxyl, amino, cyano, halogen atom, nitro or 5-to 10-membered heterocyclic group;
m is selected from O, S or N-R8，R8Selected from hydrogen, C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl or C2-6Alkynyl, said C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl and C2-6Alkynyl may optionally be substituted by C1-6Alkoxy substitution;
y is selected from N or C-R9；
X is selected from S or N-R9；
R9Selected from hydrogen, hydroxy, carboxyl, amino, cyano, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy group, (C)1-6Alkyl radical)2Amino group, C2-6Alkenyl radical, C2-6Alkynyl or 3-to 8-membered cycloalkyl.
3. The compound of claim 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester, or solvate thereof:
wherein,
R1and R3Each independently selected from hydrogen, hydroxy, amino, sulfonyl, halogen atom, C1-4Alkyl or C1-4An alkoxy group;
R2selected from hydrogen, cyano, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylthio, 5-to 10-membered heterocyclyloxy, C2-6Alkenyl radical, C2-6Alkynyl, 3-to 8-membered cycloalkyl, 5-to 6-membered heterocyclyl or 5-to 6-membered heteroaryl;
q is selected from 5-6 membered heterocyclic group, 6-10 membered heterocyclic group, 7-9 membered bridged ring group or 7-11 membered spiro ring group;
R4selected from the following groups:
(1) hydrogen, amino, hydroxy, nitro, cyano, halogen atom, C1-4Alkyl or C1-4An alkoxy group,
(2) a 7-to 10-membered bridged ring group, a 7-to 10-membered spiro ring group, a 5-to 6-membered heterocyclic group or a 6-to 10-membered heterocyclic group, which is optionally substituted with a substituent selected from the group consisting of an amino group, a hydroxyl group, a halogen atom, C1-6Alkyl radical, C1-6Alkoxy radical, C2-6Alkenyl or C2-6An alkynyl group;
n is selected from 0, 1,2 or 3, and when n is 0, R4When n is not less than 2, R4Which may be the same or different from each other,
and when Q is selected from 5-6 membered heterocyclic group, R4Can not be selected from 5-6 membered heterocyclic groups;
R5and R6Each independently selected from hydrogen, halogen atoms, C1-4Alkyl or C1-4An alkoxy group,
or R5And R6Are connected with each other to form a 5-to 6-membered heterocyclic group or a 3-to 6-membered cycloalkyl group together with the carbon atom to which they are connected;
R7selected from hydrogen, cyano, nitro, hydroxy, amino, amidino, sulfonyl, C1-6Alkyl radical, C1-6Alkoxy group, halogen atom, 3-to 6-membered cycloalkyl group, C2-6Alkenyl or C2-6An alkynyl group;
m is selected from O, S or N-R8，R8Selected from hydrogen, C1-4Alkyl or C1-4Alkoxy radical, said C1-4Alkyl radical, C1-4Alkoxy may optionally be substituted by C1-4Alkoxy substitution;
y is selected from N or C-R9；
X is selected from S or N-R9；
R9Selected from hydrogen, hydroxy, amino, halogen atoms, C1-6Alkyl radical, C1-6Alkoxy or 3-to 8-membered cycloalkyl.
4. The compound of claim 3, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester, or solvate thereof:
wherein,
R1、R2and R3Are independently selected from hydrogen or C1-4An alkyl group;
q is selected from 5-6 membered heterocyclic group, 6-10 membered heterocyclic group, 7-9 membered bridged ring group or 7-11 membered spiro ring group;
R4selected from the following groups:
(1) hydrogen, amino or C1-4An alkyl group, a carboxyl group,
(2) 7-9 membered bridged ring group or 5-6 membered heterocyclic group;
n is selected from 0 or 1, and when n is 0, R4In the absence of the presence of the agent,
and when Q is selected from 5-6 membered heterocyclic group, R4Can not be selected from 5-6 membered heterocyclic groups;
R5and R6Are independently selected from hydrogen or C1-4An alkyl group;
R7selected from hydrogen, cyano, hydroxyl, amino, methyl, ethyl or chlorine atoms;
m is selected from N-R8，R8Selected from hydrogen or C1-4An alkyl group;
y is selected from C-R9，R9Selected from hydrogen, methyl, ethyl or n-propyl;
x is selected from S.
5. The compound of claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester, or solvate thereof:
wherein,
q is selected from 4-7 membered heterocyclic group;
R4selected from 7-8 bridge ring groups;
n is selected from 1.
6. The compound of claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester, or solvate thereof:
wherein,
q is selected from 7-8 membered bridged ring groups;
R4selected from hydrogen, amino, C1-4An alkyl group or a 4-to 7-membered heterocyclic group;
n is selected from 0 or 1, and when n is 0, R4Is absent.
7. The compound of claim 1 or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester, or solvate thereof:
wherein,
q is selected from 6-10 membered heterocyclic group or 7-11 membered spiro ring group;
R4selected from hydrogen, amino or C1-4An alkyl group;
n is selected from 0 or 1, and when n is 0, R4Is absent.
8. The compound of claim 6, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, an ester thereof, or a solvate thereof, selected from:
9. a pharmaceutical composition comprising a compound according to any one of claims 1 to 8, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof, together with one or more pharmaceutically acceptable carriers and/or diluents, in any pharmaceutically acceptable dosage form.
10. The pharmaceutical composition of claim 9, further comprising one or more antineoplastic and immunosuppressive agents selected from the group consisting of methotrexate, capecitabine, gemcitabine, doxifluridine, pemetrexed disodium, pazopanib, imatinib, erlotinib, lapatinib, gefitinib, vandetanib, herceptin, bevacizumab, rituximab, trastuzumab, paclitaxel, vinorelbine, docetaxel, doxorubicin, hydroxycamptothecin, mitomycin, epirubicin, pirarubicin, bleomycin, letrozole, tamoxifen, fulvestrant, triptorelin, flutamide, leuprolide, anastrozole, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, nitrogen mustard, maflan, onconine, carboplatin, platinum, and platinum, Cisplatin, oxaliplatin, carboplatin, topotecan, camptothecin, topotecan, everolimus, sirolimus, carcinostat, 6-mercaptopurine, 6-thioguanine, azathioprine, rhzomorph D, daunorubicin, doxorubicin, mitoxantrone, bleomycin, plicamycin, or aminoglutethimide.
11. Use of a compound according to any one of claims 1 to 8, or a stereoisomer thereof, or a pharmaceutically acceptable salt, ester or solvate thereof, for the manufacture of a medicament for the treatment and/or prevention of an ALK-mediated cancer-associated disease selected from brain tumor, non-small cell lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, rectal cancer, hepatic cancer, hepatoblastoma, papillary renal cell tumor, head and neck squamous cell tumor, wilms' tumor, kidney cancer, esophageal adenocarcinoma, esophageal squamous cell carcinoma, glioma, prostate cancer, thyroid cancer, female genital tract cancer, carcinoma in situ, lymphoma, neuroblastoma, neurofibromatosis, thyroid cancer, bone cancer, skin cancer, Colon cancer, testicular cancer, small cell lung cancer, gastrointestinal stromal tumor, prostate tumor, mast cell tumor, multiple myeloma, or melanoma.
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CN1852900A (en) * 2003-09-18 2006-10-25 诺瓦提斯公司 2,4-di (phenylamino) pyrimidines useful in the treatment of proliferative disorders
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CN104804016A (en) * 2014-01-23 2015-07-29 山东轩竹医药科技有限公司 Tetra-heterocyclic-fused anaplastic lymphoma kinase inhibitor
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JP2024518783A (en) * 2021-05-12 2024-05-02 シージーンテック （スーチョウ， チャイナ） カンパニー リミテッド Compounds having ALK inhibitory activity, and their production methods and uses
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WO2023214552A1 (en) 2022-05-02 2023-11-09 セントラル硝子株式会社 Trifluoromethane sulfonylation agent composition and method for producing trifluoromethanesulfonyloxy compound or trifluoromethane sulfonyl compound

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