CN104230960B - Four-ring anaplastic lymphoma kinase inhibitor - Google Patents

Four-ring anaplastic lymphoma kinase inhibitor Download PDF

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Publication number
CN104230960B
CN104230960B CN201410157687.XA CN201410157687A CN104230960B CN 104230960 B CN104230960 B CN 104230960B CN 201410157687 A CN201410157687 A CN 201410157687A CN 104230960 B CN104230960 B CN 104230960B
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cancer
yuan
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CN104230960A (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

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

Four and ring class anaplastic lymphoma kinase ALK Alk receptor tyrosine kinase inhibitor
Technical field
The invention belongs to pharmaceutical technology field is and in particular to four and ring class anaplastic lymphoma kinase ALK Alk receptor tyrosine kinase inhibitor or it is three-dimensional Isomer or its pharmaceutically acceptable salt, ester or solvate, the preparation method of these compounds, containing these chemical combination The pharmaceutical preparation of thing and pharmaceutical composition, and this compound or its stereoisomer or its pharmaceutically acceptable salt, ester Or application in the medicine of the cancer-related diseases that preparation treatment and/or prevention are mediated by ALK for the solvate.
Background technology
Anaplastic lymphoma kinase ALK Alk receptor tyrosine kinase (Anaplastic lymphoma kinase, ALK) is receptor tyrosine kinase family Member, can raise downstream albumen by autophosphorylation, and then expresses specific gene, adjust cellular metabolism and growth.Anaplasia Property lymphom kinase is found in primary cutaneous type earliest（Anaplastic large cell lymphoma, ALCL） In, found later in nonsmall-cell lung cancer（NSCLC）In also have high expression.
The micromolecular inhibitor of ALK can affect the growth of tumor cell, plays antineoplastic action, but have and face in a large number Bed proves generation ALK inhibitor C rizotinib, easily produces drug resistance, therefore, designs and screens to Crizotinib generation The patient of drug resistance also has the secondary ALK inhibitor of good curative effect, has significant clinical meaning.
The ALK inhibitor being currently known includes Crizotinib（Pfizer）、LDK378（Novartis）、AZD-3463 （AstraZeneca）Deng.
Therefore, modified by compound structure and find the new compound structures different from Crizotinib, make great efforts to improve Its drug resistance and physicochemical property, improve druggability, such as improve the bioavailability of compound, to find active to ALK mutation Micromolecular inhibitor, for the treatment of disease clinically causing because of ALK mutation, have great importance.
Content of the invention
The present invention, to develop micromolecular inhibitor for ALK as target, has invented to treatment and/or prevention ALK mediation Cancer-related diseases have good result four and ring class anaplastic lymphoma kinase ALK Alk receptor tyrosine kinase inhibitor.Specific technical scheme be as Under：
1. formula（Ⅰ）Shown compound or its stereoisomer or its pharmaceutically acceptable salt, ester or solvation Thing：
Wherein,
A1Selected from C-R1Or N；
A2Selected from C-R2Or N；
A3Selected from C-R3Or N, and A1、A2And A3It is not simultaneously selected from N；
R1And R3Separately it is selected from hydrogen, hydroxyl, carboxyl, amino, nitro, sulfonyl, halogen atom, cyano group, C1-6Alkane Base, C1-6Alkoxyl, (C1-6Alkyl)2Amino, C2-6Thiazolinyl, C2-6Alkynyl or 3～14 yuan of cycloalkyl；
R2Selected from hydrogen, cyano group, halogen atom, C1-6Alkyl, C1-6Alkoxyl, C1-6Alkyl sulfenyl, 3～14 circle heterocycles base oxygen Base, C2-6Thiazolinyl, C2-6Alkynyl, 3～14 yuan of cycloalkyl, 3～14 circle heterocycles bases, 5～15 unit's heteroaryls or 6～14 yuan of aryl；
Q is selected from 3～8 circle heterocycles bases, 6～14 yuan and heterocyclic radical, 6～12 yuan of bridged ring bases or 6～12 yuan of volution bases；
R4Selected from following groups：
(1) hydrogen, amino, hydroxyl, nitro, cyano group, halogen atom, C1-6Alkyl, C1-6Alkoxyl, C1-6Alkyl amino, (C1-6 Alkyl)2Amino, C1-6Alkyl amino-carbonyl, hydroxyl C1-6Alkyl, hydroxyl C1-6Alkyl amino, halo C1-6Alkyl, C1-6Alkyl sulphur Acyl group, C1-6Alkyl sulfonyl-amino, amino-sulfonyl, aminosulfonylamino, C2-6Thiazolinyl or C2-6Alkynyl,
(2) 6～12 yuan of bridged ring bases being optionally substituted with a substituent, 6～12 yuan of volution bases, 3～8 circle heterocycles bases or 6～14 Unit heterocyclic radical, described substituent group is selected from amino, hydroxyl, nitro, halogen atom, carboxyl, C1-6Alkyl, C1-6Alkoxyl, C2-6Alkene Base, C2-6Alkynyl, 3～8 circle heterocycles bases or 3～8 yuan of cycloalkyl；
N is selected from 0,1,2,3,4 or 5, when n is 0, R4Do not exist, during n >=2, R4Can be identical or different,
And when Q is selected from 3～8 circle heterocycles base, R43～8 circle heterocycles bases can not be selected from；
R5And R6Separately it is selected from hydrogen, halogen atom, C1-6Alkyl, C1-6Alkoxyl, hydroxyl C1-6Alkyl, C2-6Thiazolinyl Or C2-6Alkynyl,
Or R5And R6It is connected with each other, together with the carbon atom being connected with them, form 3～14 circle heterocycles bases or 3～14 yuan of rings Alkyl；
R7Selected from hydrogen, cyano group, nitro, hydroxyl, amino, amidino groups, sulfonyl, C1-6Alkyl, C1-6Alkoxyl, halogen atom, 3 ～14 yuan of cycloalkyl, C2-6Thiazolinyl or C2-6Alkynyl, described C1-6Alkyl, C1-6Alkoxyl, 3～14 yuan of cycloalkyl, C2-6Thiazolinyl and C2-6Alkynyl can independently optionally be replaced by substituents：Hydroxyl, carboxyl, amino, cyano group, halogen atom, nitro or 3～14 Circle heterocycles base；
M is selected from O, S or N-R8, R8Selected from hydrogen, C1-6Alkyl, C1-6Alkoxyl, C2-6Thiazolinyl or C2-6Alkynyl, described C1-6Alkane Base, C1-6Alkoxyl, C2-6Thiazolinyl and C2-6Alkynyl can be optionally by C1-6Alkoxyl replaces；
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 group, halogen atom, C1-6Alkyl, C1-6Alkoxyl, (C1-6Alkane Base)2Amino, C2-6Thiazolinyl, C2-6Alkynyl or 3～14 yuan of cycloalkyl.
2. formula（Ⅰ）Shown compound or its stereoisomer or its pharmaceutically acceptable salt, ester or solvation The optimal technical scheme of thing is formula（Ⅱ）：
Wherein,
R1And R3Separately it is selected from hydrogen, hydroxyl, carboxyl, amino, nitro, sulfonyl, halogen atom, cyano group, C1-6Alkane Base, C1-6Alkoxyl, (C1-6Alkyl)2Amino, C2-6Thiazolinyl, C2-6Alkynyl or 3～8 yuan of cycloalkyl；
R2Selected from hydrogen, cyano group, halogen atom, C1-6Alkyl, C1-6Alkoxyl, C1-6Alkyl sulfenyl, 3～14 circle heterocycles base oxygen Base, C2-6Thiazolinyl, C2-6Alkynyl, 3～8 yuan of cycloalkyl, 5～6 circle heterocycles bases or 5～6 unit's heteroaryls；
Q is selected from 4～7 circle heterocycles bases, 6～12 yuan and heterocyclic radical, 7～10 yuan of bridged ring bases or 6～12 yuan of volution bases；
R4Selected from following groups：
(1) hydrogen, amino, hydroxyl, nitro, cyano group, halogen atom, C1-4Alkyl, C1-4Alkoxyl, C1-4Alkyl amino, (C1-4 Alkyl)2Amino, C1-4Alkyl amino-carbonyl, hydroxyl C1-4Alkyl, hydroxyl C1-4Alkyl amino, halo C1-4Alkyl, C1-4Alkyl sulphur Acyl group, C1-4Alkyl sulfonyl-amino, amino-sulfonyl, aminosulfonylamino, C2-6Thiazolinyl or C2-6Alkynyl,
(2) 6～10 yuan of bridged ring bases being optionally substituted with a substituent, 6～12 yuan of volution bases, 4～7 circle heterocycles bases or 6～12 Unit heterocycle, described substituent group is selected from amino, hydroxyl, halogen atom, C1-6Alkyl, C1-6Alkoxyl, C2-6Thiazolinyl or C2-6Alkynyl；
N is selected from 0,1,2 or 3, when n is 0, R4Do not exist, during n >=2, R4Can be identical or different,
And when Q is selected from 4～7 circle heterocycles base, R44～7 circle heterocycles bases can not be selected from.；
R5And R6Separately it is selected from hydrogen, halogen atom, C1-6Alkyl, C1-6Alkoxyl, hydroxyl C1-6Alkyl, C2-6Thiazolinyl Or C2-6Alkynyl,
Or R5And R6It is connected with each other, together with the carbon atom being connected with them, form 5～10 circle heterocycles bases or 3～8 yuan of cycloalkanes Base；
R7Selected from hydrogen, cyano group, nitro, hydroxyl, amino, amidino groups, sulfonyl, C1-6Alkyl, C1-6Alkoxyl, halogen atom, 3 ～8 yuan of cycloalkyl, C2-6Thiazolinyl or C2-6Alkynyl, described C1-6Alkyl, C1-6Alkoxyl, 3～8 yuan of cycloalkyl, C2-6Thiazolinyl and C2-6Alkynyl can independently optionally be replaced by substituents：Hydroxyl, carboxyl, amino, cyano group, halogen atom, nitro or 5～10 Circle heterocycles base；
M is selected from O, S or N-R8, R8Selected from hydrogen, C1-6Alkyl, C1-6Alkoxyl, C2-6Thiazolinyl or C2-6Alkynyl, described C1-6Alkane Base, C1-6Alkoxyl, C2-6Thiazolinyl and C2-6Alkynyl can be optionally by C1-6Alkoxyl replaces；
Y is selected from N or C-R9；
X is selected from S or N-R9；
R9Selected from hydrogen, hydroxyl, carboxyl, amino, cyano group, halogen atom, C1-6Alkyl, C1-6Alkoxyl, (C1-6Alkyl)2Ammonia Base, C2-6Thiazolinyl, C2-6Alkynyl or 3～8 yuan of cycloalkyl.
3. the compound as described in technical scheme 2 or its stereoisomer or its pharmaceutically acceptable salt, ester or molten Agent compound,
Wherein,
R1And R3Separately it is selected from hydrogen, hydroxyl, amino, sulfonyl, halogen atom, C1-4Alkyl or C1-4Alkoxyl；
R2Selected from hydrogen, cyano group, halogen atom, C1-6Alkyl, C1-6Alkoxyl, C1-6Alkyl sulfenyl, 5～10 circle heterocycles base oxygen Base, C2-6Thiazolinyl, C2-6Alkynyl, 3～8 yuan of cycloalkyl, 5～6 circle heterocycles bases or 5～6 unit's heteroaryls；
Q is selected from 5～6 circle heterocycles bases, 6～10 yuan and heterocyclic radical, 7～9 yuan of bridged ring bases or 7～11 yuan of volution bases；
R4Selected from following groups：
(1) hydrogen, amino, hydroxyl, nitro, cyano group, halogen atom, C1-4Alkyl or C1-4Alkoxyl,
(2) 7～10 yuan of bridged ring bases being optionally substituted with a substituent, 7～10 yuan of volution bases, 5～6 circle heterocycles bases or 6～10 Unit heterocyclic radical, described substituent group is selected from amino, hydroxyl, halogen atom, C1-6Alkyl, C1-6Alkoxyl, C2-6Thiazolinyl or C2-6Alkynes Base；
N is selected from 0,1,2 or 3, when n is 0, R4Do not exist, during n >=2, R4Can be identical or different,
And when Q is selected from 5～6 circle heterocycles base, R45～6 circle heterocycles bases can not be selected from；
R5And R6Separately it is selected from hydrogen, halogen atom, C1-4Alkyl or C1-4Alkoxyl,
Or R5And R6It is connected with each other, together with the carbon atom being connected with them, form 5～6 circle heterocycles bases or 3～6 yuan of cycloalkanes Base；
R7Selected from hydrogen, cyano group, nitro, hydroxyl, amino, amidino groups, sulfonyl, C1-6Alkyl, C1-6Alkoxyl, halogen atom, 3 ～6 yuan of cycloalkyl, C2-6Thiazolinyl or C2-6Alkynyl；
M is selected from O, S or N-R8, R8Selected from hydrogen, C1-4Alkyl or C1-4Alkoxyl, described C1-4Alkyl, C1-4Alkoxyl can be appointed Choosing is by C1-4Alkoxyl replaces；
Y is selected from N or C-R9；
X is selected from S or N-R9；
R9Selected from hydrogen, hydroxyl, amino, halogen atom, C1-6Alkyl, C1-6Alkoxyl or 3～8 yuan of cycloalkyl.
4. the compound as described in technical scheme 3 or its stereoisomer or its pharmaceutically acceptable salt, ester or molten Agent compound,
Wherein,
R1、R2And R3It is respectively selected from hydrogen or C1-4Alkyl；
Q is selected from 5～6 circle heterocycles bases, 6～10 yuan and heterocyclic radical, 7～9 yuan of bridged ring bases or 7～11 yuan of volution bases；
R4Selected from following groups：
(1) hydrogen, amino or C1-4Alkyl,
(2) 7～9 yuan of bridged ring bases or 5～6 circle heterocycles bases；
N is selected from 0 or 1, when n is 0, R4Do not exist,
And when Q is selected from 5～6 circle heterocycles base, R45～6 circle heterocycles bases can not be selected from；
R5And R6It is respectively selected from hydrogen or C1-4Alkyl；
R7Selected from hydrogen, cyano group, hydroxyl, amino, methyl, ethyl or chlorine atom；
M is selected from N-R8, R8Selected from hydrogen or C1-4Alkyl；
Y is selected from C-R9, R9Selected from hydrogen, methyl, ethyl or n-pro-pyl；
X is selected from S.
5. the compound as described in technical scheme 1 or its stereoisomer or its pharmaceutically acceptable salt, ester or molten Agent compound,
Wherein,
Q is selected from 4～7 circle heterocycles bases；
R4Selected from 7～8 yuan of bridged ring bases；
N is selected from 1.
6. the compound as described in technical scheme 1 or its stereoisomer or its pharmaceutically acceptable salt, ester or molten Agent compound,
Wherein,
Q is selected from
R4It is selected from
N is selected from 1.
7. the compound as described in technical scheme 1 or its stereoisomer or its pharmaceutically acceptable salt, ester or molten Agent compound,
Wherein,
Q is selected from 7～8 yuan of bridged ring bases；
R4Selected from hydrogen, amino, C1-4Alkyl or 4～7 circle heterocycles bases；
N is selected from 0 or 1, when n is 0, R4Do not exist.
8. the compound as described in technical scheme 1 or its stereoisomer or its pharmaceutically acceptable salt, ester or molten Agent compound,
Wherein,
Q is selected from
R4Selected from following groups：
(1) hydrogen, amino, methyl；
(2)
N is selected from 0 or 1, when n is 0, R4Do not exist.
9. the compound as described in technical scheme 1 or its stereoisomer or its pharmaceutically acceptable salt, ester or molten Agent compound,
Wherein,
Q is selected from 6～10 yuan and heterocyclic radical or 7～11 yuan of volution bases；
R4Selected from hydrogen, amino or C1-4Alkyl；
N is selected from 0 or 1, when n is 0, R4Do not exist.
10. the compound as described in technical scheme 1 or its stereoisomer or its pharmaceutically acceptable salt, ester or Solvate,
Wherein,
A1And A3Separately it is selected from CH；
A2Selected from C-R2, wherein R2Selected from hydrogen or C1-6Alkyl；
R5And R6Separately it is selected from hydrogen or C1-4Alkyl；
R7Selected from hydrogen or cyano group；
M is selected from NH；
Y is selected from CH；
X is selected from S.
11. compounds as described in technical scheme 1 or its stereoisomer or its pharmaceutically acceptable salt, ester or Solvate, described compound is selected from：
12. compounds as described in any one of technical scheme 1-11 or its stereoisomer or it is pharmaceutically acceptable Salt, ester or solvate and one or more pharmaceutical carrier and/or diluent pharmaceutical composition, can make and pharmaceutically may be used The arbitrary dosage form accepting.
13. pharmaceutical compositions as described in technical scheme 12 are it is characterised in that also can contain one or more antitumor agent And immunosuppressant, described antitumor agent and immunosuppressant are selected from methotrexate, Capecitabine, gemcitabine, deoxygenate fluorine Uridnine, pemetrexed disodium, pazopanib, imatinib, erlotinib, Lapatinib, gefitinib, ZD6474, He Sai Spit of fland, bevacizumab, Rituximab, Herceptin, paclitaxel, vinorelbine, docetaxel, doxorubicin, hydroxy-camptothecin Alkali, mitomycin, epirubicin, Pirarubicin, bleomycin, letrozole, tamoxifen, fulvestrant, music score of Chinese operas Rayleigh, fluorine he Amine, leuprorelin, Anastrozole, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, nitrogen Mustard, L-Sarcolysinum, Chlorambucil, carboplatin, cisplatin, oxaliplatin, network platinum, Topotecan, camptothecine, topotecan, everolimuses, west Luo Mosi, special cancer are fitted, Ismipur, 6- thioguanine, azathioprine, rhzomorph D, daunorubicin, amycin, mitoxantrone, Bleomycin A5, plicamycin or aminoglutethimide.
14. compounds as described in any one of technical scheme 1-11 or its stereoisomer or it is pharmaceutically acceptable Salt, ester or solvate preparation for treat and/or prevent ALK mediation cancer-related diseases medicine in application, The related disease of described cancer be selected from cerebroma, lung cancer in non-cellule type, squamous cell cancer, bladder cancer, gastric cancer, ovarian cancer, Peritoneal cancer, cancer of pancreas, breast carcinoma, head and neck cancer, cervical cancer, carcinoma of endometrium, rectal cancer, hepatocarcinoma, hepatoblastoma, mamillary Renal cell carcinoma, incidence squamous cytoma, nephroblastoma, renal carcinoma, adenocarcinoma of esophagus, esophageal squamous cell carcinoma, glioma, Carcinoma of prostate, thyroid carcinoma, female reproductive tract cancer, cancer in situ, lymphoma, neuroblastoma, neurofibromatosiss, thyroid Cancer, osteocarcinoma, skin carcinoma, colon cancer, carcinoma of testis, small cell lung cancer, gastrointestinal stromal tumor, tumor of prostate, mast cell tumor, Multiple myeloma or melanoma.
Detailed Description Of The Invention
" halogen atom " of the present invention refers to fluorine atom, chlorine atom, bromine atoms, atomic iodine etc..
" C of the present invention1-6Alkyl " represents the alkyl containing 1-6 carbon atom of straight or branched, including such as " C1-4 Alkyl ", " C1-3Alkyl " etc., such as methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl, isobutyl group, sec-butyl, the tert-butyl group, positive penta Base, isopentyl, 2- methyl butyl, neopentyl, 1- ethyl propyl, n-hexyl, isohesyl, 3- methyl amyl, 2- methyl amyl, 1- Methyl amyl, 3,3- dimethylbutyl, 2,2- dimethylbutyl, 1,1- dimethylbutyl, 1,2- dimethylbutyl, 1,3- diformazan Base butyl, 2,3- dimethylbutyl, 2- ethyl-butyl, 1,2- dimethyl propyl etc.." C of the present invention1-4Alkyl " refers to above-mentioned “C1-6In alkyl " example, carbon number is the instantiation of 1-4.
" C of the present invention1-6Alkoxyl " refers to above-mentioned " C1-6The group that alkyl " is connected with other structures by oxygen atom, As methoxyl group, ethyoxyl, propoxyl group, 1- methyl ethoxy, butoxy, 1- methyl propoxyl group, 2- methyl propoxyl group, 1,1- diformazan Base oxethyl, amoxy, 1- methylbutoxy group, 2- methylbutoxy group, 3- methylbutoxy group, 1,1- dimethyl propylene epoxide, 1,2- Dimethyl propylene epoxide, 2,2- dimethyl propylene epoxide, 1- ethylpropoxy, hexyloxy, 1- methyl amoxy, 2- methyl amoxy, 3- methyl amoxy, 4- methyl amoxy, 1,1- dimethyl butyrate epoxide, 1,2- dimethyl butyrate epoxide, 1,3- dimethyl butyrate epoxide, 2,2- dimethyl butyrate epoxide, 2,3- dimethyl butyrate epoxide, 3,3- dimethyl butyrate epoxide, 1- ethyl-butoxy, 2- ethyl-butoxy, 1,1,2- trimethyl propoxyl group, 1,2,2- trimethyl propoxyl group, 1- ethyl -1- methyl propoxyl group and 1- Ethyl-2-Methyl third oxygen Base etc.." C of the present invention1-4Alkoxyl " refers to above-mentioned " C1-6In alkoxyl " example, carbon number is the instantiation of 1-4.
" C of the present invention2-6Thiazolinyl " refer to the carbon number containing at least one double bond be 2-6 straight or branched or The thiazolinyl of ring-type, including such as " C2-4Thiazolinyl ", " C2-3Thiazolinyl ", " C3-6Cycloalkenyl group " etc., such as vinyl, acrylic, 2- propylene Base, cyclobutenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, cyclopentenyl, cyclopentadienyl group, cyclohexene Base and cyclohexadienyl etc..Double bond is optionally cis and trans.
" C of the present invention2-6Alkynyl " refers to the straight or branched that the carbon number containing at least one three key is 2-6 Alkynyl, including such as " C2-6Alkynyl ", " C2-4Alkynyl ", " C2-3Alkynyl " etc., such as acetenyl, propinyl, butynyl, pentyne Base, hexin base etc..
" C of the present invention1-6Alkyl amino ", " C1-6Alkyl sulfenyl ", " C1-6Alkyl amino-carbonyl ", " C1-6Alkyl sulfonyl Base ", " C1-6Alkyl sulfonyl-amino " refers to above-mentioned " C respectively1-6Alkyl " passes through amino, sulfenyl, amino carbonyl, sulfonyl, sulphonyl The group that base amino is connected with other structures.Term " C1-4Alkyl sulphonyl " refers to above-mentioned " C1-4Alkyl " pass through sulfonyl and its The group that his structure is connected.
" hydroxyl C of the present invention1-6Alkyl ", " halo C1-6Alkyl " refers to hydroxyl respectively, halogen atom replaces above-mentioned " C1-6 One or more hydrogen atoms on alkyl ", and the group being connected with other structures by alkyl.Term " hydroxyl C1-4Alkyl ", " halo C1-4Alkyl " refers to hydroxyl respectively, halogen atom replaces above-mentioned " C1-4One or more hydrogen atoms on alkyl ", and pass through The group that alkyl is connected with other structures, wherein " halogen atom " are as mentioned before.
" (C of the present invention1-6Alkyl)2Amino " refers in amino atom that two energy are substituted by above-mentioned " C1-6Alkyl " Replaced, and the group being connected with other structures by amino.
" 3～14 yuan of cycloalkyl " of the present invention refers to that the paraffin section of 3～14 carbon atoms removes a hydrogen atom Derivative cyclic alkyl, including 3～8 yuan of cycloalkyl, 6～14 yuan and ring cycloalkyl.
3～8 yuan of cycloalkyl, refer to that the paraffin section of 3～8 carbon atoms removes cyclic alkyl derived from a hydrogen atom, Including such as " 3～6 yuan of cycloalkyl ", " 3～5 yuan of cycloalkyl ", " 5～6 yuan of cycloalkyl " etc., the example includes but is not limited to：Ring Propyl, Tetramethylene. base, Pentamethylene. base, cyclohexyl, cycloheptyl alkyl, cyclooctane base, methyl cyclopropane base, dimethylcyclopropane Base, methyl cyclobutane base, dimethylcyclobutane base, methyl cyclopentane base, dimethylcyclopentane base, methyl cyclohexane alkyl, diformazan Butylcyclohexane base etc.." 3～6 yuan of cycloalkyl " of the present invention refers to contain 3～6 in above-mentioned " 3～8 yuan of cycloalkyl " example The instantiation of individual carbon atom.
6～14 yuan and ring cycloalkyl, refer to that shared two adjacent carbon are former each other by two or more circuluses 6～14 cyclic group that son is formed, the example includes but is not limited to：Bicyclo- [3.1.0] hexyl, bicyclo- [4.1.0] heptan Alkyl, bicyclo- [2.2.0] hexyl, bicyclo- [3.2.0] heptane base, bicyclo- [4.2.0] octyl, octahydro pentalene base, Octahydro -1H- indenyl, decahydronaphthalene naphthyl, ten tetrahydrochysene phenanthryl, bicyclic [3.1.0] hex- 2- thiazolinyl, bicyclic [4.1.0] hept- 3- thiazolinyl, Bicyclic [3.2.0] hept- 3- thiazolinyl, bicyclic [4.2.0] octyl- 3- thiazolinyl, 1,2,3,3a- tetrahydrochysene pentalene base, 2,3,3a, 4, 7,7a- hexahydro -1H- indenyl, 1,2,3,4,4a, the octahydro naphthyl of 5,6,8a-, 1,2,4a, 5,6,8a- hexahydro naphthyl, 1,2, 3,4,5,6,7,8,9,10- decahydro phenanthryl etc..
" hetero atom " of the present invention refers to N, O, C (O), S, SO and/or SO2Deng preferably N, O, S, more preferably N, O.
" 3～14 circle heterocycles base " of the present invention refers to containing one or more heteroatomic 3～14 cyclic group, including 3～8 circle heterocycles bases and 6～14 yuan and heterocyclic radical.
3～8 circle heterocycles bases, refer to containing 3～8 annular atoms（Wherein at least contains a hetero atom）Monocyclic heterocycles Base.Instantiation include but are not limited to 2,5- dihydro-thiophene base, 4,5- pyrazoline base, 3,4- dihydro -2H- pyranose, 5, 6- dihydro -4H-1,3- piperazine base, aziridine base, azetidinyl, Thietane base, tetrahydrofuran base, tetrahydrochysene Pyrrole radicals, imidazolidinyl, pyrazolidinyl, tetrahydrofuran base, 1,4- dioxane base, 1,3- dioxane base, 1, 3- dithian base, morpholinyl, piperazinyl etc..The present invention " 3～6 circle heterocycles base ", " 4～7 circle heterocycles base ", " 5～6 yuan Heterocyclic radical " refer to above-mentioned " 3～8 circle heterocycles base " medium ring atomic number be 3～6 yuan, 4～7 yuan, 5～6 yuan of instantiation.
6～14 yuan and heterocyclic radical, refer to containing 6～14 annular atoms（Wherein at least contains a hetero atom）By two or Two or more circulus shares the condensed cyclic structure that two adjacent atoms couple together formation, such as benzo 3～8 circle heterocycles each other Structure that the structure that base is formed, 3～8 circle heterocycles bases 3～8 circle heterocycles bases are formed etc., instantiation includes but is not limited to： Former etc. the circulus arbitrarily commutable hydrogen of replacement The group that son is formed.
" 6～12 yuan and heterocyclic radical " of the present invention, " 6～10 yuan and heterocyclic radical " refer to above-mentioned " 6～14 yuan and heterocyclic radical " Medium ring atomic number is 6～12 yuan, 6～10 yuan of instantiation.
" 5～15 unit's heteroaryl " of the present invention finger ring atom is that 5～15 yuan of inclusion is one or more heteroatomic Cyclic aromatic groups, including 5～8 yuan of single heteroaryls and 8～15 yuan of thick heteroaryls.
5～8 yuan of single heteroaryls, including but not limited to pyrrole radicals, imidazole radicals, pyrazolyl, 1,2,3-triazoles base, 1,2,4- tri- Oxazolyl, pyridine radicals, furyl, thienyl,Oxazolyl, differentOxazolyl, thiazolyl, isothiazolyl, 1,2,3- thiadiazolyl group, 1,2, 4- thiadiazolyl group, 1,3,4- thiadiazolyl group, 1,2,3-Di azoly, 1,2,4Di azoly, 1,2,5-Di azoly, 1,2,3- Triazine radical, 1,2,4- triazine radical, tetrazole radical,Triazolyl, 2H-1,2Piperazine base, 4H-1,2-Piperazine base, 6H-1,2-Piperazine base, 2H-1,3-Piperazine base, 4H-1,3-Piperazine base, 6H-1,3-Piperazine base, 2H-1,4-Piperazine base, 4H-1,4-Piperazine base, differentPiperazine Base, pyridazinyl, pyrimidine radicals and pyrazinyl etc.；
8～15 yuan of thick heteroaryls, refer to containing 8～15 annular atoms（Wherein at least contains a hetero atom）By two or Two or more hetero-aromatic ring shares the condensed cyclic structure that two adjacent atoms couple together formation, including but not limited to benzo furan each other Mutter base, isobenzofuran-base, benzothienyl, indyl, isoindolyl, quinolyl, isoquinolyl, indolizine base, indazolyl, phthalein Piperazine base, quinoxalinyl, quinazolyl, benzodiazine base, benzisoxaOxazolyl, benzoPiperazine base, benzimidazolyl, pyrido Pyridine radicals, pyrazolo [3,4-b] pyridine radicals, purine radicals, acridinyl and ton base etc..Preferably " 8～12 yuan of thick heteroaryls ", " 8～ 10 unit's heteroaryls ", " 9～10 yuan of thick heteroaryls " refer to above-mentioned " 8～15 yuan of thick heteroaryls " example medium ring atomic number be respectively 8～ 12 yuan, 8～10 yuan, 9～10 yuan of instantiation.
It is 5 that term " 5～10 unit's heteroaryl ", " 5～6 unit's heteroaryl " refer to above-mentioned " 5～15 unit's heteroaryl " medium ring atomic number ～10 yuan, 5～6 yuan of instantiation.
" 6～14 yuan of aryl " of the present invention refers to the cyclic nonaromatics that annular atom is 6～14 yuan of carbon atoms Remove the monovalent moiety that hydrogen atom obtains, including 6～8 yuan of aryl and 8～14 yuan of fused ring aryl.6～8 yuan of aryl include phenyl, Cyclooctatetraenyl etc..8～14 yuan of fused ring aryl refer to share two adjacent carbon atoms each other by two or more aromatic rings The condensed ring group being formed, at least one ring is the cyclic group of all undersaturated aromatic rings, including 8～14 yuan all not Saturation condensed ring carbon aryl, such as naphthyl, anthryl, phenanthryl etc., also include 8～14 yuan of fractional saturation fused ring aryl, such as benzo 3～8 First cycloalkyl, instantiation such as 2,3- dihydro -1H- indenyl, 1H- indenyl, 1,2,3,4- tetralyls, Isosorbide-5-Nitrae-ihydro naphthyl etc.. Term " 6～8 yuan of aryl " refers to the instantiation that above-mentioned " 6～14 yuan of aryl " medium ring atomic number is 6～8 yuan.
" 6～12 yuan of bridged ring bases " of the present invention refers to that any two ring shares containing of two non-conterminous atoms formation 6～12 carbon atoms or/and heteroatomic structure.Including such as " 6～10 yuan of bridged ring bases ", " 6～8 yuan of bridged ring bases ", " 7 ～10 yuan of bridged ring bases ", " 7～9 yuan of bridged ring bases ", " 7～8 yuan of bridged ring bases ", " 8 yuan of bridged ring bases ", " 6～12 yuan of bridge heterocyclic radicals ", " 6 ～10 yuan of bridge heterocyclic radicals ", " 6～8 yuan of bridge heterocyclic radicals ", " 7～10 yuan of bridge heterocyclic radicals ", " 7～9 yuan of bridge heterocyclic radicals ", " 7～8 yuan of bridges Heterocyclic radical " and " 8 yuan of bridge heterocyclic radicals " etc..The example includes but is not limited to：
Deng.
" 6～12 yuan of volution bases " of the present invention refer to that at least two rings share that atom formed containing 6～ 12 carbon atoms or/and heteroatomic structure.Including such as " 7-12 unit volution base ", " 7-11 unit volution base ", " 7～10 First volution base ", " 7～9 yuan of volution bases ", " 7～8 yuan of volution bases ", " 8 yuan of volutions ", " 6-12 unit spiro heterocyclic radical ", " 7-12 unit spiral shell Heterocyclic radical ", " 7-11 unit spiro heterocyclic radical ", " 7-10 unit spiro heterocyclic radical ", " 7-9 unit spiro heterocyclic radical ", " 7-8 unit spiro heterocyclic radical " and " 8 First spiro heterocyclic radical " etc..The example includes but are not limited to：
Deng.
Present invention also offers the three of above-claimed cpd kinds of preparation methoies, but it is not limited only to following methods, reaction equation As follows：
Preparation method 1：
Reactions steps：
The preparation of step 1 intermediate 1,2,7
Intermediate 1,2 and 7 is bought respectively or is prepared.
The preparation of step 2 intermediate 3
Intermediate 1 and appropriate intermediate 2 are dissolved in appropriate solvent（Such as trifluoroacetic acid）In, it is heated to reflux to having reacted Finish, cooling, remove solvent, separate to obtain intermediate 3 through proper method.
The preparation of step 3 intermediate 4
Intermediate 3 is dissolved in appropriate solvent, add appropriate DDQ (DDQ), room temperature or be heated to react Finish, separate to obtain intermediate 4 through proper method.
The preparation of step 4 intermediate 5
By intermediate 5 and appropriate pyridine hydrochloride mix homogeneously or appropriate solvent dissolving, it is heated to reaction and finishes, through suitable When method separates to obtain intermediate 5.
The preparation of step 5 intermediate 6
Intermediate 5 is placed in appropriate solvent, adds appropriate trifluoromethanesulfanhydride anhydride, stir and finish to reaction, through suitably square Method separates to obtain intermediate 6.
The preparation of step 6 intermediate 8
Intermediate 6 and appropriate intermediate 7 are placed in appropriate solvent, are heated to reaction and finish, separate through proper method pure Change to obtain intermediate 8.
The preparation of step 7 intermediate 9
Intermediate 8 and appropriate Dai Si-Martin reagent are added to appropriate mixed solution（Such as dichloromethane and dimethyl are sub- Sulfone）In, it is stirred at room temperature（Such as 2 hours）.Separate to obtain intermediate 9 through proper method.
Step 8 formula（Ⅰ）The preparation of compound
Intermediate 9, indium chloride are dissolved in appropriate solvent（Such as N,N-dimethylformamide）In, nitrogen is protected, and room temperature is stirred Mix（Such as 3 hours）.Add appropriate sodium cyanoborohydride, be stirred overnight at room temperature, separate to obtain formula through proper method（Ⅰ）Chemical combination Thing.
Preparation method 2：
Reactions steps：
The preparation of step 1 intermediate 6
With reference to step 1-6 in above-mentioned preparation method 1.
The preparation of step 2 intermediate 9
Intermediate 7 and intermediate 8 are dissolved in appropriate solvent, add appropriate sodium cyanoborohydride, be stirred at room temperature to reaction Finish, separate through proper method and obtain intermediate 9.
Step 3 formula（Ⅰ）The preparation of compound
Intermediate 6 and intermediate 9 are dissolved in appropriate solvent, add suitable alkali（Such as diisopropyl ethyl amine）Microwave adds Heat（Such as 2 hours）, separate through proper method and obtain product.
Preparation method 3：
The preparation of step 1 intermediate 2
Intermediate 1 is dissolved in appropriate solvent (such as acetonitrile), room temperature adds N- bromo-succinimide, stirs to having reacted Entirely, add water and be quenched, sucking filtration obtains intermediate 2.
The preparation of step 2 intermediate 11
With reference to step 2- step 8 in above-mentioned preparation method 1.
The preparation of step 3 intermediate 12
By intermediate 11, triisopropylsilyl acetylene, alkali（Such as cesium carbonate）With suitable part（Such as X-Phos bis- hexamethylene Base phosphorus -2', 4', 6'- tri isopropyl biphenyl）And palladium catalyst（For example double (acetonitrile) palladium chloride）It is added in acetonitrile, nitrogen Protection is lower to heat（Such as 80 DEG C）Reaction, cooling（For example to room temperature）, add solvent（Such as ethyl acetate）, saturated sodium-chloride water Solution, is layered to obtain organic faciess, and organic faciess obtain intermediate 12 through proper method purification.
The preparation of step 4 intermediate 13
Intermediate 12 is dissolved in appropriate solvent（Such as oxolane）, add tetrabutyl ammonium fluoride, be stirred at room temperature to reaction Completely, add water and be quenched, organic solvent（Such as ethyl acetate）Extraction, merges organic faciess, obtains intermediate through appropriate separation purification 13.
Step 5 formula（Ⅱ）The preparation of compound
Intermediate 13 is dissolved in appropriate solvent（Such as methanol）, add appropriate palladium carbon, be passed through hydrogen, be stirred at room temperature to reaction Completely, filter palladium carbon, filtrate is concentrated, obtain the present invention through appropriate separation purificationFormula（Ⅱ）Compound.
In reaction equation, A1、A2、A3、M、X、Y、Q、R1、R3、R4、R5、R6、R7With n as defined hereinabove, X' represents halogen Plain atom.
The formula of the present invention（I）Shown arbitrary compound pharmaceutically acceptable salt refer to by pharmaceutically acceptable, Non-toxic alkali or the standby salt of processed with acid, including acylate, inorganic acid salt, organic alkali salt, inorganic base salts.Acylate includes first Acid, acetic acid, benzenesulfonic acid, benzoic acid, p-methyl benzenesulfonic acid, camphorsulfonic acid, citric acid, methanesulfonic acid, ethyl sulfonic acid, propane sulfonic acid, fumaric acid, Gluconic acid, glutamic acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, glactaric acid, pamoic acid, pantothenic acid, succinic acid, wine The salt of stone acid etc..Inorganic acid salt includes the salt of hydrobromic acid, hydrochloric acid, nitric acid, sulphuric acid, phosphoric acid etc..
Organic alkali salt includes primary, secondary and tertiary amine, and substituted amine includes naturally occurring replacement amine, cyclammonium and basic ion and exchanges Resin, selected from glycine betaine, caffeine, choline, N, N '-dibenzyl-ethylenediamin, diethylamine, 2- Diethylaminoethanol, 2- dimethylamine Base ethanol, ethanolamine, ethylenediamine, N- ethyl-morpholine, N-ethylpiperidine, meglumine, glucosamine, Hai Baming, isopropyl Amine, methylglucosamine, morpholine, piperazine, piperidines, procaine, purine, theobromine, triethylamine, trimethylamine, tripropyl amine (TPA), ammonia fourth three The salt of alcohol etc..Native amino hydrochlorate for example glycine, alanine, L-Valine, leucine, isoleucine, nor-leucine, tyrosine, Cystine, cysteine, Methionine, proline, hydroxyproline, histidine, ornithine, lysine, arginine, serine etc. Salt.Inorganic base salts include the salt of ammonium and lithium, sodium, potassium, calcium, magnesium, zinc, barium, aluminum, ferrum, ketone, ferrous iron, manganese, bivalent manganese etc..
Claimed formula（I）" stereoisomer " of compound refers to work as formula（I）Compound exist chiral centre, During double bond etc., produced all stereoisomers, including enantiomer, diastereomer, cis-trans-isomer, mutually make a variation Structure body, geometric isomer, epimer and its mixture, are included in the scope of the invention.
The formula of the present invention（I）If the raceme that shown arbitrary compound synthesis obtain, required enantiomer-pure Compound can be obtained by the method for chiral separation：Can be by having the chromatography of chiral stationary phase（Image height compacting is standby Liquid phase, supercritical fluid chromatography）.Chirality padding includes but is not limited to：Chiralcel OJ-H, Chiralpak AD-H, Chiralpak IA, Chiralpak AS-H.
Formula of the present invention（I）" ester " of compound refers to, works as formula（I）When compound has carboxyl, with alcohol, ester can occur The ester changed reaction and formed, works as formula（I）When compound has hydroxyl, ester can be occurred with organic acid, mineral acid, acylate etc. The ester changed reaction and formed.Ester, under conditions of acid or alkali exist, can occur hydrolysis to generate acid or alcohol accordingly.
Compound shown in logical formula I or its stereoisomer or its pharmaceutically acceptable salt or ester can be " molten Agent compound " form.In the case that solvate is hydrate, hydration can complete in preparation process or can utilize The hygroscopicity of original anhydrous product is gradually carried out.
Further requirement protection of the present invention includes above-mentioned formula（I）Shown arbitrary compound or its stereoisomer, Or its pharmaceutically acceptable salt, ester or solvate and the drug regimen of one or more pharmaceutical carrier and/or diluent Thing, can make pharmaceutically acceptable arbitrary dosage form.Being applied in modes such as oral, parenteral, rectum or transpulmonary administration is needed Want the patient of this treatment.During for oral administration, can be made into the solid preparation of routine, such as tablet, capsule, pill, granule Agent etc.；May be made as oral liquid, such as oral solution, oral suspensionses, syrup etc..When making oral formulations, can To add suitable filler, binding agent, disintegrating agent, lubricant etc..During for parenteral, can be made into injection, including Injection, injectable sterile powder and concentrated solution for injection.When making injection, can be using the routine side in existing pharmaceutical field Method produces, and when preparing injection, can be added without additives, and the property also dependent on medicine adds suitable additives.For During rectally, can be made into suppository etc..During for transpulmonary administration, can be made into inhalant or spray etc..
Further requirement protection of the present invention includes formula recited above（I）Arbitrary compound or its stereoisomer or Its pharmaceutically acceptable salt of person, ester or solvate and the medicine group of other one or more antitumor agent and immunosuppressant Compound.Described antitumor agent and immunosuppressant, such as anti-metabolism, include but are not limited to methotrexate, Capecitabine, Ji His shore, Doxifluridine, pemetrexed disodium of west；Growth factor receptor inhibitors class, includes but are not limited to pazopanib, she horse replaces Buddhist nun, erlotinib, Lapatinib, gefitinib, ZD6474；Antibody class, includes but are not limited to Herceptin, bevacizumab； Targeting class, includes but are not limited to Herceptin, bevacizumab, Rituximab, Herceptin；Mitotic inhibitor class, Include but are not limited to paclitaxel, vinorelbine, docetaxel, doxorubicin, hydroxy camptothecin, mitomycin, epirubicin, Pirarubicin, bleomycin；Antitumor hormoness, include but are not limited to letrozole, tamoxifen, fulvestrant, the music score of Chinese operas auspicious Woods, flutamide, leuprorelin, Anastrozole；Alkylating agents, include but are not limited to ifosfamide, busulfan, ring phosphinylidyne Amine, carmustine, nimustine, semustine, chlormethine, L-Sarcolysinum, Chlorambucil；Metal platinum class, include but are not limited to carboplatin, Cisplatin, oxaliplatin, network platinum；Topoisomerase enzyme inhibitor, includes but are not limited to Topotecan, camptothecine, topotecan；Exempt from Epidemic disease suppresses class, includes but are not limited to everolimuses, Sirolimus, special cancer are fitted；Purine analogue, selected from Ismipur, 6- Thioguanine, azathioprine；Antibioticses, selected from rhzomorph D, daunorubicin, amycin, mitoxantrone, plicamycin；Adrenal gland Cortex inhibitor class, selected from aminoglutethimide.
Present invention also offers formula of the present invention（I）Shown compound or its stereoisomer or its pharmaceutically may be used Application in the medicine of preparation treatment and/or the cancer-related diseases of prevention ALK mediation for salt, ester or the solvate accepting, The related disease of described cancer is selected from cerebroma, pulmonary carcinoma, lung cancer in non-cellule type, squamous cell cancer, bladder cancer, gastric cancer, ovum Nest cancer, peritoneal cancer, cancer of pancreas, breast carcinoma, head and neck cancer, cervical cancer, carcinoma of endometrium, rectal cancer, hepatocarcinoma, renal carcinoma, esophageal gland Cancer, esophageal squamous cell carcinoma, solid tumor, non-Hodgkin lymphoma, thyroid carcinoma, female reproductive tract cancer, cancer in situ, lymphoma, Histocytic lymphoma, neurofibromatosiss, osteocarcinoma, skin carcinoma, colon cancer, carcinoma of testis, small cell lung cancer, Gastrointestinal Stromal Tumor, tumor of prostate, mast cell tumor, multiple myeloma, melanoma, glioma, astrocytoma, neuroblast Tumor, sarcoma etc..
The compounds of this invention has advantages below：
（1）Formula（I）Compound or its stereoisomer or its pharmaceutically acceptable salt, ester or solvation Thing has excellent ALK and NCI-H3122, the inhibitory activity of Karpas-299, NCI-H2228 cell；
（2）Formula（I）Compound or its stereoisomer or its pharmaceutically acceptable salt, ester or solvation Thing shows good pharmacokinetic property, persistent, and bioavailability is high；
（3）The compounds of this invention preparation process is simple, medicine purity is high, and steady quality is it is easy to carry out large-scale industry life Produce.
The compounds of this invention beneficial effect is expanded on further below by way of the experiment of external zymetology inhibitory activity, but should not be by this It is interpreted as that the compounds of this invention only has following beneficial effect.
The external zymetology activity experiment of experimental example 1 the compounds of this invention
Test sample：The compounds of this invention 1-3,5-11 and 13, its chemical name and preparation method ask for an interview the system of each compound Standby embodiment.The implication that the abbreviation of following middle experiments is representative is as follows：
DMSO：Dimethyl sulfoxide
DTT：Dithiothreitol, DTT
SEB：Enzyme catalyst buffer solution
ATP：Adenosine triphyosphate
ALK：Anaplastic lymphoma kinase ALK Alk receptor tyrosine kinase
SA-XL665：The donor of marked by streptavidin
2.5 ×, 5 ×, 10 × "×" therein：Times
Experimental technique：
ALK kinase buffer liquid is prepared：
Take the MgCl that appropriate mother liquid concentration is 1000mM respectively2, the DTT of SEB, 100mM of 2500nM, 5 × enzyme buffer liquid, It is added in ultra-pure water so that ultimate density is respectively：5mM, 25nM, 1mM, 1 × enzyme buffer liquid, mix, stand-by.
2.5 × need testing solution is prepared：
The 1mM storing solution of compound is prepared：Weigh Compound is appropriate (concrete sample weighting amount please see table) respectively, adds appropriate DMSO dissolves, and mixes, standby.
Take 1mM storing solution respectively, make, with DMSO dilution, the solution that concentration is 200 μM, as mother solution.Will be above-mentioned with DMSO Mother solution three times stepwise dilution makes a series of solution of concentration, and then each concentration uses ALK kinase buffer liquid to dilute 80 times respectively, Make each 2.5 × need testing solution, concentration is respectively：2500nM、833.33nM、277.78nM、92.59nM、30.86nM、 10.29nM、3.43nM、1.14nM、0.38nM、0.13nM、0.04nM.
Various other preparation of reagents：
5 × ALK kinase solution required for being prepared respectively with ALK kinase buffer liquid, 5 × substrate solution, 5 × ATP solution, Standby.
ALK zymetology is reacted：
1）It is separately added into 2.5 × need testing solution that 4 μ L prepare, 2 μ L prepare in corresponding hole in 384 orifice plates 5 × ALK kinase solution, 25 DEG C are incubated 10 minutes.
2）Corresponding Kong Zhongzai is separately added into 5 × substrate solution that 2 μ L prepare and 5 × ATP that 2 μ L prepare is molten Liquid, starts enzyme reaction, and 25 DEG C are incubated 30 minutes.
Zymetology detects：
Prepare the SA-XL665 of desired concn with detection buffer (detection buffer), then with isopyknic cheese Histidine kinase antibody mixes, and is separately added into this antibody-solutions that 10 μ L prepare, terminating reaction in corresponding hole.25 DEG C of incubations 1h.
Microplate reader 665nm/615nm read plate.
IC50：Calculate suppression ratio (%)=(maximum-sample ratio)/(maximum-minima) × 100, using Graph Prisim software carries out curve fitting, and draws IC50Value.
Maximum：It is not added with the positive control of compound, minima：Not enzyme-added negative control.
Experimental result and conclusion：
The external zymetology inhibitory activity of table 1 the compounds of this invention
From table 1, the compounds of this invention has good inhibitory activity to ALK kinases, can be used for treating and kinases phase Disease, the kinase mediated disease of particularly ALK or the patient's condition closed, has significant clinical meaning.
The cell in vitro activity experiment of experimental example 2 the compounds of this invention
Test sample：The compounds of this invention 1,2,3,5,6 and 8-12, its chemical name and preparation method are shown in the system of each compound Standby embodiment.
The implication that the abbreviation of following middle experiments is representative is as follows：
rpm：Rpm
DMSO：Dimethyl sulfoxide
MTS：Thiazole bromide blue tetrazolium
RPMI1640：1640 culture medium (RPMI:Roswell Park Memorial Institute)
500 ×, 1000 ×, 10 × "×" therein：Times
Compound 1 experimental technique：
（One）NCI-H3122 and Karpas-299 cell：
（1）Prepared by cell：
With containing 10% hyclone, 100U/ml penicillin, 100mg/ml streptomycin RPMI-1640 culture medium, 5% CO2, 37 DEG C of conditions incubator in, cultured cells is to 80% fusion, standby.
（2）Inoculating cell：
With trypsin digestion cell, 1000rpm centrifugation 4min, remove supernatant, with the RPMI-1640 containing 10% hyclone Culture medium is suspended again, adjusts cell density, takes this cell suspension 90 μ L to be inoculated in 96 orifice plates, obtains final cell close Spend for Karpas-299：1500/hole；NCI-H3122：2000/hole；Then in 5%CO2, cultivate 24h in 37 DEG C of incubators.
（3）Add test sample：
（3.1）Compound 1 solution compound method：Weigh Compound 14.99mg is appropriate, adds appropriate DMSO to dissolve and is used in combination DMSO gradient dilution makes a series of mother solution of concentration（1000 × need testing solution）, then dilute 100 times of this mother solution with culture medium Obtain 10 × need testing solution, take this solution 10 μ L respectively, be added in the corresponding hole of 96 orifice plates, obtain need testing solution final Concentration is：10μM、2.5μM、625nM、156nM、39nM、9.8nM、2.5nM.
（3.2）Control wells are arranged：
Vehicle controls：0.1%DMSO.
Cell controls：Inoculating cell, is not added with compound.
Blank：Culture medium, instrument returns to zero.
（3.3）96 orifice plates are put 37 DEG C, 5%CO272h is cultivated in incubator.
（4）Detection:
（4.1）MTS detection method：Applicable cell：NCI-H3122 and Karpas-299
1. willReagent room temperature in single solution 96 hole cell proliferation detecting kit places 90min.
2. add in each test hole of 96 orifice platesThe mono- solution reagent of AQueous 20 μ L.
3. 96 orifice plates are put 5%CO2, cultivate 2h in 37 DEG C of incubators.
4. microplate reader Detection wavelength 490nm is set, reads result.
（5）Result treatment：
IC50Calculate：Cell survival rate (%)=(sample value-blank value)/(maximum-blank value) × 100, adopts Graph prisim software carries out curve fitting, and draws IC50Value.
Maximum：Be not added with compound solubilization matchmaker cell controls, blank value：Blank value.
（Two）NCI-H2228 cell：
（1）Prepared by cell：
With the RPMI-1640 culture medium containing 10% hyclone, in 5%CO2, 37 DEG C of conditions incubator in, cultured cells To 80% fusion, standby.
（2）Inoculating cell：
With trypsin digestion cell, 1000rpm centrifugation 4min, remove supernatant, with the RPMI-1640 containing 10% hyclone Culture medium is suspended again, adjusts cell density 2 × 104Individual/mL, takes this cell suspension 100 μ L to be inoculated in 96 orifice plates, obtains Final cell density is：2000 cells/well.
（3）Add test sample：
（3.1）Compound 1 solution is prepared：Weigh Compound 13.14mg, adds appropriate DMSO, dissolving, mixes, by solution Use DMSO gradient dilution, obtain a series of solution of each concentration, standby.
99 μ L culture medium are taken to be added separately in each hole of 96 orifice plates, then take the solution of the above-mentioned variable concentrations preparing 1 μ L is added in corresponding hole so that the ultimate density of compound 1 is：10000nM,3333.33nM,1111.11nM, 370.37nM,123.45nM,41.15nM,13.71nM,4.57nM,1.52nM,0.50nM.
（3.2）This 96 orifice plate is put 5%CO2, 37 DEG C of conditions incubator in cultivate 96h.
（4）Detection：
CTG detection method：
1. culture medium in every for 96 orifice plates hole is removed, add new culture medium 100 μ L, equilibrium at room temperature 30min.
2. add in each test hole of 96 orifice platesReagent 60 μ L.
3. 96 orifice plates are vibrated with micropore plate oscillator lucifuge and mix 2min, make cell cracking.
4. by 96 orifice plate lucifuge incubated at room 10min, make the optical signal value of generation stable.
5. microplate reader reads result under luminescence pattern.
（5）Result treatment：
Carried out curve fitting using Graph prisim software, draw IC50Value.
Other parts compound experimental technique of the present invention：
（One）NCI-H3122, Karpas-299 cell：
（1）Prepared by cell：
With containing 10% hyclone, 100U/ml penicillin, 100mg/ml streptomycin RPMI-1640 culture medium, 5% CO2, 37 DEG C of conditions incubator in, cultured cells is to 80% fusion, standby.
（2）Inoculating cell：
With trypsin digestion cell, 1000rpm centrifugation 4min, remove supernatant, with the RPMI-1640 containing 2.5% hyclone Culture medium is suspended again, adjusts cell density, takes this cell suspension 90 μ L to be inoculated in 96 orifice plates, obtains final cell density For 3000/hole；Then in 5%CO2, cultivate 24h in 37 DEG C of incubators.
（3）Add test sample：
（3.1）Need testing solution is prepared
Need testing solution compound method one：Weigh test sample 22.93mg, add appropriate DMSO to dissolve and use DMSO gradient dilute Release a series of mother solution making concentration（200 × need testing solution）, then with culture medium dilute this mother solution 20 obtain 10 again × for examination Product solution, takes this solution 10 μ L respectively, is added in the corresponding hole of 96 orifice plates, obtains test sample 2 solution ultimate density and is：10μ M、2.5μM、625nM、156nM、39nM、9.8nM、2.5nM.
Need testing solution compound method two：Weigh test sample respectively appropriate（Concrete sample weighting amount please see table）, add appropriate DMSO dissolves and makes a series of mother solution of concentration respectively with DMSO gradient dilution（1000 × need testing solution）, then use respectively Culture medium dilutes this mother solution 100 and obtains 10 × need testing solution again, takes this solution 10 μ L respectively, is added to the corresponding hole of 96 orifice plates In, obtaining each need testing solution ultimate density is：10μM、2.5μM、625nM、156nM、39nM、9.8nM、2.5nM.
（3.2）Control wells are arranged：
Vehicle controls：0.1%DMSO or 0.2%DMSO, 0.5%DMSO.
Cell controls：Inoculating cell, is not added with compound.
Blank：Culture medium, instrument returns to zero.
（3.3）96 orifice plates are put 37 DEG C, 5%CO272h is cultivated in incubator.
（4）Detection:
MTS detection method：
1. willReagent room temperature in single solution 96 hole cell proliferation detecting kit places 90min.
2. add in each test hole of 96 orifice platesThe mono- solution reagent of AQueous 20 μ L.
3. 96 orifice plates are put 5%CO2, cultivate 40min in 37 DEG C of incubators.
4. microplate reader Detection wavelength 490nm is set, reads result.
（5）Result treatment：
IC50Calculate：Cell survival rate (%)=(sample value-blank value)/(maximum-blank value) × 100, adopts Graph prisim software carries out curve fitting, and draws IC50Value.
Maximum：Be not added with compound solubilization matchmaker cell controls, blank value：Blank value.
（Two）NCI-H2228 cell：
（1）Prepared by cell：
With the RPMI-1640 culture medium containing 10% hyclone, in 5%CO2, 37 DEG C of conditions incubator in, cultured cells To 80% fusion, standby.
（2）Inoculating cell：
With trypsin digestion cell, 1000rpm centrifugation 4min, remove supernatant, with containing 2.5% hyclone（Compound 2,9 Use 10% hyclone with 12）RPMI-1640 culture medium be again suspended, adjust cell density 2 × 104Individual/mL, takes this cell Suspension 100 μ L is inoculated in 96 orifice plates, obtains final cell density and is：2000 cells/well.
（3）Add test sample：
（3.1）Need testing solution is prepared：Weigh test sample respectively appropriate（Concrete sample weighting amount please see table）, add appropriate DMSO, dissolving, mix, solution DMSO gradient dilution obtains a series of solution of each concentration, standby.
99 μ L culture medium are taken to be added separately in each hole of 96 orifice plates, then take the solution of the above-mentioned variable concentrations preparing 1 μ L is added in corresponding hole so that the ultimate density of the compounds of this invention is：10000nM、2500nM、625nM、 156.25nM、39.06nM、9.77nM、2.44nM、0.61nM、0.15nM.The ultimate density of comparison medicine is：9800nM、 2450nM、612.5nM、153.1nM、38.28nM、9.57nM、2.39nM、0.60nM、0.15nM.
（3.2）Control wells are arranged：
Vehicle controls：0.5%DMSO.
Cell controls：Inoculating cell, is not added with compound.
Blank：Culture medium, instrument returns to zero.
Taxol control：Add 1000nM paclitaxel, inoculating cell, as minimum comparison.
（3.3）This 96 orifice plate is put 5%CO2, 37 DEG C of conditions incubator in cultivate 96h.
（4）Detection：
CTG detection method：
1. culture medium in every for 96 orifice plates hole is removed 80 μ L, equilibrium at room temperature 30min.
2. add in each test hole of 96 orifice platesReagent 60 μ L.
3. 96 orifice plates are vibrated with micropore plate oscillator lucifuge and mix 2min, make cell cracking.
4. by 96 orifice plate lucifuge incubated at room 10min, make the optical signal value of generation stable.
5. microplate reader reads result under luminescence pattern.
（5）Result treatment：
Computational methods one：IC50Calculate：Cell inhibitory rate (%)=(ODSolvent- ODCompound)/(ODSolvent- ODpositive) × 100, Carried out curve fitting using Graph prisim software, draw IC50Value.
ODSolvent：Be not added with compound solubilization matchmaker cell controls, ODpositive：Add the cell pair of 1000nM paclitaxel According to.
Computational methods two：IC50Calculate：Cell inhibitory rate (%)=(ODSolvent- ODCompound)/(ODSolvent- ODNegative) × 100, adopt Graph prisim software carries out curve fitting, and draws IC50Value.
ODSolvent：Be not added with compound solubilization matchmaker cell controls, ODNegative：Culture medium blank
Experimental result：
The cell inhibitory activity of table 2 the compounds of this invention
From table 2, the compounds of this invention is respectively provided with well to cell NCI-H3122, Karpas-299 and NCI-H2228 Inhibitory activity, can be used for treating the kinase mediated disease of ALK or the patient's condition, there is significant clinical meaning.
The Pharmacokinetics in Rat experiment of experimental example 3 the compounds of this invention
Test sample：The compounds of this invention 11, self-control, its chemical name and preparation method are shown in that the preparation of compound 11 is implemented Example.
Animal subject：Male SD rat, 3/route of administration/test sample；Rat body weight 200- used by the compounds of this invention 230g/ is only.
Prepared by need testing solution：
Animal IV administration group：
Weigh Compound 112.87mg, adds DMSO101.46 μ L, ultrasonic dissolution, adds 40%HP- β-CD（Hydroxy propyl-Beta- Cyclodextrin）1.015mL, it is vortexed and mixes, be incubated 20min in 50 DEG C of thermostat water baths, add sterilized water for injection 3.957mL, is vortexed and mixes, and filtration obtains final product.
40%HP- β-CD solution compound method：Take HP- β-CD4g to add sterilized water for injection 10mL, ultrasonic dissolution, be vortexed Mix, obtain final product.
Animal PO administration group：
Weigh Compound 113.06mg, adds 2%HPC+0.1%Tween805.586mL, puts in tissue grinder, The rotating speed of 600rpm/min, is uniformly dispersed and obtains final product.
2%HPC+0.1%Tween80 solution compound method：Weigh hydroxypropyl cellulose（HPC）20g, is slowly added to In 1000mL water, continuously stirred make fully to dissolve, add 1mL Tween80, mix homogeneously, obtain final product.
Experimental technique
Need testing solution is carried out intravenous injection administration by administration（iv）, dosage is 1mg/kg, administered volume 2mL/kg； Test sample gastric infusion（po）, dosage is 2mg/kg, administered volume 4mL/kg.
Blood sampling
Compound 11IV administration after 0.083,0.25,0.5,1,2,4,6,8,24,32,48h, PO administration after 0.167,0.5, 1st, 2,4,6,8,24,32,48h carries out tail vein blood, each time point take 100 μ L about whole blood, in the high speed of 8000rpm It is centrifuged 6min separated plasma, blood plasma is frozen in -80 DEG C of refrigerators in centrifuge.Blood plasma must be made in 30 minutes after blood collection Standby.
Plasma sample analysis
Plasma sample adopts liquid-liquid extraction method：Take 20 μ L blood plasma, add the MTBE (tert-butyl group first that 600 μ L contain CH5424802 Ether) solution（10ng/mL）, 1500 revs/min of vortex 10min, then 12000 revs/min of centrifugation 5min, take supernatant 300 μ L, Dry up under a nitrogen, with 300 μ L acetonitriles:Water（7:3, V/V）Redissolve, be vortexed and mix, LC-MS/MS analyzes.
Table 3.1 P of Rats K evaluation result (iv:1mg/kg)
Table 3.2 P of Rats K evaluation result (po:2mg/kg)
AUClastRepresent area under the drug-time curve 0 → t
CL represents clearance rate
VssRepresent apparent steady state distribution volume
T1/2Represent the half-life
TmaxRepresent blood medicine peak time
CmaxRepresent blood peak concentration of drug
F% represents absolute bioavailability
From the experimental result of table 3.1 and table 3.2, the compounds of this invention 11 pharmacokinetic property is good.
Specific embodiment
The specific embodiment of form by the following examples, makees further specifically to the above of the present invention Bright.But this scope being interpreted as the above-mentioned theme of the present invention should not be only limitted to following examples.All based on the above of the present invention The technology realized belongs to the scope of the present invention.
It is defined as follows representated by following abbreviations：
EA：Ethyl acetate
PE：Petroleum ether
DCM：Dichloromethane
NMP：N-Methyl pyrrolidone
DMSO：Dimethyl sulfoxide
THF：Oxolane
TFA：Trifluoroacetic acid
TBAF：Tetrabutyl ammonium fluoride
DMF：Dimethylformamide
X-Phos：2- dicyclohexyl phosphine -2', 4', 6'- tri isopropyl biphenyl
DDQ：The chloro- 5,6- dicyano -1,4- benzoquinone of 2,3- bis-
NBS：N- bromo-succinimide
Embodiment 1：7- (4- (3- oxa- -8- azabicyclo [3.2.1] octane -8- base) piperidin-1-yl) -5,5- diformazan The preparation of base -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 1)
(1) preparation of 2- (4- bromothiophene -2- base) -1,3- dioxolanes
4- bromothiophene -2- formaldehyde (19.1g, 0.10mol) and pyrovinic acid (1.0g, 0.010mol) are dissolved in ethylene glycol (200mL), in, it is heated to reflux and divides water 6 hours.Resulting solution rotary evaporation removes solvent, and residue is through silica gel column chromatography (stone Oily ether:Ethyl acetate=10:1) separate to obtain 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- formylthiophene -3- base) boric acid
By 2- (4- bromothiophene -2- base) -1,3- dioxolanes (16g, 0.068mol) and triisopropyl borate ester (14.1g, 0.075mol) it is dissolved in dry THF (oxolane) (150mL), nitrogen is protected, and is slowly added to n-BuLi (positive fourth at -78 DEG C Base lithium) (30mL, 2.5M), stir 1 hour after completion of dropping, be warmed to room temperature, add 10% hydrochloric acid, stir 1 hour.Use acetic acid Ethyl ester extracts, anhydrous sodium sulfate drying, and rotary evaporation removes solvent and obtains product (7.5g, yield 71%).
1H-NMR(400MHz,CD3OD)δ:9.92(s,1H),8.33(s,1H),8.16(s,1H).
(3) preparation of 1- (5- formylthiophene -3- base) hydrazine -1,2- diformazan tert-butyl acrylate
By (5- formylthiophene -3- base) boric acid (7.5g, 0.0481mol) and azo di tert butyl carbonate (11.08g, 0.0481mol) and the copper acetate (0.44g, 2.4mmol) of catalytic amount is added in oxolane (150mL), it is stirred at room temperature 16 Hour.Rotary evaporation removes solvent, residue silica gel column chromatography (petroleum ether:Ethyl acetate=10:1) separate to obtain product (11.7g, yield 71%).
(4) preparation of 1- (5- cyano thiophene -3- base) hydrazine -1,2- diformazan tert-butyl acrylate
By 1- (5- formylthiophene -3- base) hydrazine -1,2- diformazan tert-butyl acrylate (11.7g, 0.0342mol) and oxammonium hydrochloride. (4.8g, 0.0691mol) is dissolved in pyridine (60mL), and 90 DEG C are stirred 10 minutes, are cooled to room temperature, add acetic anhydride (20.9g, 0.205mol), it is again heated to 80 DEG C to stir 1 hour, add ethyl acetate (200mL) to reactant liquor, molten with 10% hydrochloric acid successively Liquid, water, saturated nacl aqueous solution washing, rotary evaporation removes solvent, silica gel column chromatography (ethyl acetate:Petroleum ether=1:10) divide From product (10.4g, yield 90%).
(5) preparation of 4- diazanyl thiophene -2- carbonitrile hydrochloride
1- (5- cyano thiophene -3- base) hydrazine -1,2- diformazan tert-butyl acrylate (10.4g, 0.031mol) is dissolved in dioxane (65mL) in, add concentrated hydrochloric acid (39mL), be heated to 80 DEG C and stir 2 hours, rotary evaporation removes solvent afforded crude material (5.4g).
(6) 7- methoxyl group -5,5- dimethyl -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN Preparation
By 4- diazanyl thiophene -2- carbonitrile hydrochloride (5.4g, 0.031mol) and 7- methoxyl group -1,1- dimethyl -3,4- two Hydrogen naphthalene -2- (1H) -one (6.3g, 0.031mol) is dissolved in trifluoroacetic acid (150mL), is heated to reflux 40 minutes, cooling, will react Liquid is poured in ethyl acetate and unsaturated carbonate potassium solution, point liquid, and organic faciess are washed through saturated nacl aqueous solution, is dried, rotary evaporation Remove solvent, silica gel post separation (ethyl acetate:Petroleum ether=1:5) obtain product (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) 7- methoxyl group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole - The preparation of 2- formonitrile HCN
By 7- methoxyl group -5,5- dimethyl -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (1.6g, 5.2mmol) is dissolved in THF (oxolane) (50mL) and the mixed solvent of water (5mL), cools down under ice-water bath, adds DDQ (DDQ) (1.8g, 7.9mmol), is then naturally warmed to room temperature and stirs 30 minutes, and ethyl acetate extracts, The organic faciess merging, through unsaturated carbonate potassium solution and water washing, are dried, and rotary evaporation removes solvent, residue silica gel column chromatography (petroleum ether:Ethyl acetate=1:1) separate and obtain product (0.52g, yield 31%).
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) 7- hydroxyl -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- The preparation of formonitrile HCN
By 7- methoxyl group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole - 2- formonitrile HCN (0.52g, 1.6mmol) and pyridine hydrochloride (2.96g, 25.6mmol) mix homogeneously, microwave heating, to 170 DEG C, stirs Mix 15 minutes, after finishing, mixture is placed in water, ethyl acetate extracts, the organic faciess of merging are dried, and rotary evaporation removes molten Agent obtains crude product (437mg).
(9) 2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- The preparation of base triflate
By 7- hydroxyl -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- Formonitrile HCN crude product (437mg) is dissolved in pyridine (10mL), is slowly added to trifluoromethanesulfanhydride anhydride (440mg, 1.56mmol) under ice-water bath, It is stirred at room temperature after adding 3 hours, reactant liquor is poured in water, add ethyl acetate extraction, the organic faciess of merging are through saturated common salt Water washing, rotary evaporation removes solvent, obtains crude product (462mg).
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) 7- (4- hydroxy piperidine -1- base) -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno The preparation of [3,2-b] indole -2- formonitrile HCN
By 2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- Base triflate (440mg, 1mmol) and 4- hydroxy piperidine (1.0g, 0.01mol) are dissolved in N-Methyl pyrrolidone (30mL) In, 120 DEG C of oil baths 1 hour.Cooling, adds ethyl acetate (200mL), is washed with saturated sodium-chloride water solution, rotary evaporation removes Solvent is gone to obtain crude product (400mg).
Molecular formula：C22H21N3O2S molecular weight：391.49LC-MS(m/z)：392.0[M+H]+
(11) 5,5- dimethyl -10- oxo -7- (4- oxo-piperidine -1- base) -5,10- dihydro -4H- benzo [f] thieno The preparation of [3,2-b] indole -2- formonitrile HCN
By 7- (4- hydroxy piperidine -1- base) -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thiophene [3, 2-b] indole -2- formonitrile HCN (400mg, 1.02mmol) and Dai Si-Martin reagent (520mg, 1.2mmol) be added to dichloromethane (20mL), and in the mixed solution of dimethyl sulfoxide (8mL), it is stirred at room temperature 2 hours.Add dichloromethane (200mL), use saturation Sodium bicarbonate solution and saturated common salt water washing, take organic faciess to be dried, rotary evaporation removes solvent, obtains crude product (230mg).
Molecular formula：C22H19N3O2S molecular weight：389.47LC-MS(m/z)：390.0[M+H]+
(12) 7- (4- (3- oxa- -8- azabicyclo [3.2.1] octane -8- base) piperidin-1-yl) -5,5- dimethyl -10- The preparation of oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 5,5- dimethyl -10- oxo -7- (4- oxo-piperidine -1- base) -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (230mg, 0.59mmol), indium chloride (130mg, 0.59mmol) and 3- oxa- -8- azabicyclo [3.2.1] octane hydrochloride (177mg, 1.18mmol) is dissolved in DMF (15mL), and nitrogen is protected, and room temperature is stirred Mix 3 hours.Add sodium cyanoborohydride (74mg, 1.18mmol), be stirred overnight at room temperature, add ethyl acetate (200mL), with satisfying Wash with sodium-chloride water solution, preparative hplc separates to obtain end-product (20mg, yield 7%).
Molecular formula：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).
Embodiment 27- (4- (8- oxa- -3- azabicyclo [3.2.1] octane -3- base) piperidin-1-yl) -5,5- dimethyl - The preparation of 10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 2)
(1) 2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- The preparation of base triflate
Will according to embodiment 1 (1st) step the-the (8) one step preparation method preparation 7- hydroxyl -5,5- dimethyl -10- oxo - 5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (10.1g, 32.8mmol) is dissolved in anhydrous pyridine (50mL) in, Deca trifluoromethanesulfanhydride anhydride (27.7g, 98.2mmol) under ice bath, completion of dropping in 20min, it is back to room temperature and continue Reaction 4h.Add water (200mL) and ethyl acetate (200mL), carry out a point liquid extraction, after organic faciess concentrate, residue is through post layer Analysis purification (PE:EA=3:1), obtain product (7.9g, yield 55%).
(2) 7- (4- hydroxy piperidine -1- base) -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno The preparation of [3,2-b] indole -2- formonitrile HCN
By 2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- Base triflate (400mg, 0.91mmol), 4- hydroxy piperidine (551mg, 5.45mmol) are added to N-Methyl pyrrolidone (5mL) in, microwave heating to 130 DEG C of reaction 2h.Reaction finishes, and reactant liquor is poured in water (30mL), separates out solid and carries out Decompression sucking filtration, gained crude product is through column chromatography purification (DCM:MeOH=40:1) obtain product (200mg, yield 56%).
(3) 5,5- dimethyl -10- oxo -7- (4- oxo-piperidine -1- base) -5,10- dihydro -4H- benzo [f] thieno The preparation of [3,2-b] indole -2- formonitrile HCN
By 7- (4- hydroxy piperidine -1- base) -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (200mg, 0.51mmol) is dissolved in DCM (10mL), adds DMSO (4mL) and triethylamine (4mL) add, more in batches and under sulfur trioxide pyridine complex (488mg, 3.1mmol) room temperature, react 12h.System adds DCM Carry out extraction point liquid with water, after organic faciess concentrate, through column chromatography (DCM:MeOH=50:1) purification obtains product (150mg, yield 76%).
(4) 7- (4- (8- oxa- -3- azabicyclo [3.2.1] octane -3- base) piperidin-1-yl) -5,5- dimethyl -10- The preparation of oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 5,5- dimethyl -10- oxo -7- (4- oxo-piperidine -1- base) -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (150mg, 0.39mmol) is dissolved in THF (5mL) and methanol (5mL), then is separately added into HOAC (0.5mL) with 8- oxa- -3- azabicyclo [3.2.1] octane hydrochloride (105mg, 0.7mmol), sustained response 1h under room temperature. Then add sodium cyanoborohydride (73mg, 1.16mmol) under ice bath in batches, be warmed to room temperature sustained response 4h.Water quenching is added to go out Reaction, rotary evaporation removes organic solvent, adds EA and water to carry out extraction point liquid, after organic faciess concentrate, inverted preparative hplc Purification obtains product (40mg, yield 21%).
Molecular formula：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).
Embodiment 37- (4- (3- oxa- -8- azabicyclo [3.2.1] octane -8- base) piperidin-1-yl) -8- ethyl -5,5- The preparation of dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 3)
(1) preparation of 7- methoxyl group -1,1- dimethyl -3,4- dihydronaphthalene -2 (1H) -one
By 7- methoxyl group -3,4- dihydronaphthalene -2 (1H) -one (50g, 0.28mol), iodomethane (100g, 0.70mol) dissolving In THF (500mL), add the aqueous solution of 50% KOH (125g, 1.1mol), flow back 2 hours, stopped reaction, add acetic acid Ethyl ester (2L) extracts at twice, is dried, and rotary evaporation removes solvent, adds methanol (500mL), water (250mL), crystallizes to obtain product (45g, yield 79%).
(2) preparation of 6- bromo- 7- methoxyl group -1,1- dimethyl -3,4- dihydronaphthalene -2 (1H) -one
7- methoxyl group -1,1- dimethyl -3,4- dihydronaphthalene -2 (1H) -one (30g, 0.15mol) is added to acetonitrile (600mL) in, it is stirred at room temperature down, adds NBS (28.5g, 0.16mol) to stir 4 hours, be added in 1L water to system and stir 30 Minute, sucking filtration is dried, and obtains product (38g, yield 91%).
(3) synthesis of 4- bromothiophene -2- formonitrile HCN
4- bromothiophene -2- formaldehyde (600g, 3.14mol) and oxammonium hydrochloride. (438g, 6.30mol) are added to pyridine (5L) In, it is heated to 90 DEG C after 10 minutes and is down to room temperature, Deca acetic anhydride (1940g, 19.0mol) is heated to 80 DEG C, react 1 hour, It is poured in water (20L), stir 30 minutes, sucking filtration, dry to obtain product (564g, yield 95%).
(4) synthesis of 4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolanes -2- base) thiophene -2- formonitrile HCN
By 4- bromothiophene -2- formonitrile HCN (564g, 3.0mol), duplex pinacol borate (805g, 3.16mol), [1,1'- Double (diphenylphosphine) ferrocene] palladium chloride (56g), potassium acetate (885g, 9.02mol) dioxane (9L), be averagely added to 2 In individual four-hole bottle, under nitrogen protection, it is heated to 90 DEG C, react 15h, stopped reaction, rotary evaporation removes solvent, and residue is through post Chromatography (petroleum ether：Ethyl ester=10:1) purification obtains product (426g, yield 60%).
(5) preparation of di-t-butyl 1- (5- cyano thiophene -3- base) hydrazine -1,2- dicarboxylic acid esters
By 4- (4,4,5,5- tetramethyl -1,3,2- two heterocycle pentaborane -2- base) thiophene -2- formonitrile HCN (426g, 1.8mol), Schweinfurt green (18g, 0.09mol), di-t-butyl (Z)-diazene -1,2- dicarboxylic acid esters (621g, 2.7mol) is added to THF (6L) in, 16h is stirred at room temperature, rotary evaporation removes solvent, and residue is through column chromatography (petroleum ether：Ethyl acetate=10:1) purification Obtain product (396g, yield 65%).
(6) preparation of 4- diazanyl thiophene -2- carbonitrile hydrochloride
Di-t-butyl 1- (5- cyano thiophene -3- base) hydrazine -1,2- dicarboxylic acid esters (396g, 1.17mol) is dissolved in dioxy In six rings (3L), add 80 DEG C of stirring 2h of concentrated hydrochloric acid (1584mL), rotary evaporation removes solvent and obtains product (140g, yield 68%).
(7) 8- bromo- 7- methoxyl group -5,5- dimethyl -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- The preparation of formonitrile HCN
By bromo- for 6- 7- methoxyl group -1,1- dimethyl -3,4- dihydronaphthalene -2 (1H) -one (28.5g, 0.1mol), 4- diazanyl thiophene Fen -2- carbonitrile hydrochloride (17.6g, 0.1mol) is dissolved in trifluoroacetic acid (200mL), is heated to reflux 40 minutes, is cooled to room temperature, Add the saturated aqueous solution of ethyl acetate and sodium carbonate, point liquid, organic faciess are washed with saturated sodium bicarbonate aqueous solution, are dried, and remain Excess is through column chromatography (petroleum ether：Ethyl acetate=1:1) purification obtains product (17.6g, yield 45%).
(8) 8- bromo- 7- methoxyl group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] The synthesis of indole -2- formonitrile HCN
By bromo- for 8- 7- methoxyl group -5,5- dimethyl -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- first Nitrile (17.6g, 0.045mol) is dissolved in THF (200mL), add water (20mL), under ice-water bath add DDQ (20.7g, 0.09mol), addition finishes, and is warming up to room temperature, stirs 15h, and reaction terminates, and rotary evaporation removes solvent, residue column chromatography (petroleum ether：Ethyl acetate=1:1) isolate and purify to obtain product (9.5g, yield 52%).
(9) 8- bromo- 7- hydroxyl -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] Yin The synthesis of diindyl -2- formonitrile HCN
By bromo- for 8- 7- methoxyl group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] Indole -2- formonitrile HCN (9g, 0.02mol) and pyridine hydrochloride (45g), mix homogeneously, 170 DEG C of microwave reactions 15 minutes, will mix Thing is dissolved in water, is extracted with ethyl acetate, organic faciess anhydrous sodium sulfate drying, and rotary evaporation removes solvent and obtains product (5.6g, product Rate 72%).
(10) 8- bromo- 2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] The preparation of indole -7- base triflate
By bromo- for 8- 7- hydroxyl -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] Yin Diindyl -2- formonitrile HCN (5.0g, 12.9mmol) is dissolved in pyridine (100mL), be slowly added under ice bath trifluoromethanesulfanhydride anhydride (4.0g, 14.2mmol), it is stirred at room temperature after adding 3 hours, add ethyl acetate extraction, and washed with water, saturated sodium-chloride water solution successively Wash, anhydrous sodium sulfate drying, rotary evaporation removes solvent and obtains product (5.2g, yield 78%).
(11) the bromo- 7- of 8- (4- hydroxy piperidine -1- base) -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] The preparation of thieno [3,2-b] indole -2- formonitrile HCN
By bromo- for 8- 2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] Yin Diindyl -7- base triflate (1.0g, 1.9mmol) and 4- hydroxy piperidine (1.92g, 19mmol) are added in NMP (20mL), 120 DEG C of oil baths 5 hours, add ethyl acetate (100mL), use water successively, and saturated sodium-chloride water solution washs, and anhydrous sodium sulfate is done Dry, residue is through silica gel column chromatography (PE:EA=1:1) obtain product (700mg, yield 77%).
(12) 8- bromo- 5,5- dimethyl -10- oxo -7- (4- oxo-piperidine -1- base) -5,10- dihydro -4H- benzo [f] The preparation of thieno [3,2-b] indole -2- formonitrile HCN
By bromo- for 8- 7 (4- hydroxy piperidine -1- base) -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thiophene And [3,2-b] indole -2- formonitrile HCN (640mg, 1.36mmol) is dissolved in DCM (8mL) and DMSO (4mL), add triethylamine (3mL), Lower addition sulfur trioxide pyridine (2.16g, 13.7mmol) is stirred at room temperature, adds after finishing, be stirred overnight at room temperature, TLC detection is anti- Completely, after rotary evaporation removes solvent, should add water and stir out precipitation, sucking filtration obtains filter cake, filter cake is through silica gel column chromatography (dichloromethane Alkane：Methanol=30:1) purification obtains product (500mg, yield 78%).
(13) 7- (4- (3- oxa- -8- azabicyclo [3.2.1] octane -8- base) piperidin-1-yl) -8- bromo- 5,5- diformazan The preparation of base -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By bromo- for 8- 5,5- dimethyl -10- oxo -7- (4- oxo-piperidine -1- base) -5,10- dihydro -4H- benzo [f] thiophene Fen simultaneously [3,2-b] indole -2- formonitrile HCN (500mg, 1.07mmol) and 3- oxa- -8- azabicyclo [3.2.1] octane hydrochloride (319mg, 213mmol) is dissolved in DMF (10mL), adds Indium-111 chloride (237mg, 1.07mmol), is stirred overnight under room temperature, plus Enter sodium cyanoborohydride (134mg, 2.13mmol), after being stirred at room temperature 5 hours, add water (50mL) to stir out precipitation, sucking filtration, filter Cake is through column chromatography (dichloromethane：Methanol=20:1) purification obtains product (510mg, yield 84%).
(14) 7- (4- (3- oxa- -8- azabicyclo [3.2.1] octane -8- base) piperidin-1-yl) -5,5- dimethyl -10- Oxo -8- ((triisopropylsilyl) acetenyl) -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN Preparation
By 7- (4- (3- oxa- -8- azabicyclo [3.2.1] octane -8- base) piperidin-1-yl) -8- bromo- 5,5- dimethyl - 10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (510mg, 0.9mmol), triisopropyl Silicon substrate acetylene (246mg, 1.35mmol), cesium carbonate (1.31g, 4.02mmol) and X-Phos (129mg, 0.27mmol) and double (acetonitrile) palladium chloride (23mg, 0.09mmol) is added in acetonitrile (20mL), and lower 80 DEG C of nitrogen protection is reacted 4 hours, and TLC examines Measured reaction completely, is down to room temperature, adds ethyl acetate (100mL) and saturated sodium-chloride water solution (50mL), is layered to obtain organic faciess, Aqueous phase is extracted with ethyl acetate (50mL), merges organic faciess, anhydrous sodium sulfate drying, rotary evaporation removes solvent afforded crude material (650mg) not purified it is directly used in next step.
(15) 7- (4- (3- oxa- -8- azabicyclo [3.2.1] octane -8- base) piperidin-1-yl) -8- acetenyl -5,5- The preparation of dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 7- (4- (3- oxa- -8- azabicyclo [3.2.1] octane -8- base) piperidin-1-yl) -5,5- dimethyl -10- oxygen Generation -8- ((triisopropylsilyl) acetenyl) -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN crude product (650mg) it is dissolved in THF (10mL), adds TBAF (380mg, 1.45mmol), after being stirred at room temperature 1 hour, add water and acetic acid second The each 50mL of ester, is layered to obtain organic faciess, organic phase washed with water six times, anhydrous sodium sulfate drying, rotary evaporation removes solvent, remaining Thing is through column chromatography (dichloromethane：Methanol=20:1) purification obtains product (350mg).
(16) 7- (4- (3- oxa- -8- azabicyclo [3.2.1] octane -8- base) piperidin-1-yl) -8- ethyl -5,5- two The preparation of methyl isophthalic acid 0- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 7- (4- (3- oxa- -8- azabicyclo [3.2.1] octane -8- base) piperidin-1-yl) -8- acetenyl -5,5- two Methyl isophthalic acid 0- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (100mg, 0.20mmol) is dissolved in In THF (6mL) and methanol (4mL), add Pd/C (60mg), be stirred at room temperature under hydrogen environment 2 hours, sucking filtration, filtrate rotation is steamed Send out and remove solvent through column chromatography (dichloromethane：Methanol=15:1) purification obtains end-product (70mg, yield 69%).Molecular formula： 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).
Embodiment 45,5- dimethyl -7- (8- morpholine -3- azabicyclo [3.2.1] octane -3- base) -10- oxo -5,10- The preparation of dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 4)
(1) preparation of tert-butyl group 8- morpholine -3- azabicyclo [3.2.1] octane -3- formic acid esters
By tert-butyl group 8- oxo -3- azabicyclo [3.2.1] octane -3- formic acid esters (600mg, 2.66mmol), it is dissolved in In THF (5mL) and methanol (5mL), then it is separately added into acetic acid (0.5mL) and morpholine (255mg, 2.93mmol), continue under room temperature Reaction 1 hour.Then add sodium cyanoborohydride (336mg, 5.3mmol) under ice bath in batches, be back to room temperature sustained response 4 Hour.Add water quenching to go out reaction, rotary evaporation removes organic solvent, add ethyl acetate and water to carry out extraction and divide a liquid, gained has Machine phase rotary evaporation removes solvent, and gained crude product is through column chromatography (PE:EA=10:1) purification obtains product (408mg, yield 52%)..
(2) preparation of 4- (3- azabicyclo [3.2.1] octane -8- base) morpholine
Tert-butyl group 8- morpholine -3- azabicyclo [3.2.1] octane -3- formic acid esters (400mg, 1.35mmol) is dissolved in In DCM (20mL), add TFA (10mL), sustained response 2 hours under room temperature.System rotary evaporation removing solvent is added Saturated sodium bicarbonate aqueous solution adjusts pH value to 9, and rotary evaporation removes solvent, and residue is through column chromatography (DCM:MeOH:Et3N= 40:1:0.1) obtain product (215mg, yield 81%).
(3) 5,5- dimethyl -7- (8- morpholine -3- azabicyclo [3.2.1] octane -3- base) -10- oxo -5,10- two The preparation of hydrogen -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
Will be according to the 2- cyano group -5,5- dimethyl -10- oxo -5,10- two of embodiment 2 (1st) one step preparation method preparation Hydrogen -4H- benzo [f] thieno [3,2-b] indole -7- base triflate (161mg, 0.37mmol), 4- (3- azabicyclo [3.2.1] octane -8- base) morpholine (215mg, 1.1mmol) is dissolved in N-Methyl pyrrolidone (10mL), adds N, N- bis- Wopropyl ethyl amine (472mg, 3.7mmol), the lower 120 DEG C of sustained responses of microwave 2 hours.Reaction finishes, and reactant liquor is poured into water (20mL), in, system separates out solid decompression sucking filtration, and gained solid is through column chromatography (DCM：MeOH=20:1) purification obtains end-product (35mg, yield 20%).
Molecular formula：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).
Embodiment 55,5- dimethyl -7- (3- morpholine -8- azabicyclo [3.2.1] octane -8- base) -10- oxo -5,10- The preparation of dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 5)
(1) 7- (3- hydroxyl -8- azabicyclo [3.2.1] octane -8- base) -5,5- dimethyl -10- oxo -5,10- two The preparation of hydrogen -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
Will be according to the 2- cyano group -5,5- dimethyl -10- oxo -5,10- two of embodiment 2 (1st) one step preparation method preparation Hydrogen -4H- benzo [f] thieno [3,2-b] indole -7- base triflate (400mg, 0.91mmol), 8- azabicyclo [3.2.1] octane -3- alcohol (693mg, 5.45mmol) is added in N-Methyl pyrrolidone (5mL), the lower 130 DEG C of reactions 2 of microwave Hour.Reaction finishes pours in 30mL water by this system, and system separates out solid and carries out the sucking filtration that reduces pressure, and gained crude product is through column chromatography (DCM:MeOH=40:1) purification obtains product (200mg, yield 53%).
(2) 5,5- dimethyl -10- oxo -7- (3- oxo -8- azabicyclo [3.2.1] octane -8- base) -5,10- two The preparation of hydrogen -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 7- (3- hydroxyl -8- azabicyclo [3.2.1] octane -8- base) -5,5- dimethyl -10- oxo -5,10- dihydro - 4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (200mg, 0.48mmol) is dissolved in 10mL DCM, adds DMSO (4mL) with triethylamine (4mL), then add reaction 12 under sulfur trioxide pyridine complex (458mg, 2.9mmol) room temperature in batches Hour.System adds DCM and water to carry out extraction point liquid, and gained organic faciess rotary evaporation removes solvent, through column chromatography (DCM: MeOH=50:1) purification obtains product (159mg, yield 80%).
(3) 5,5- dimethyl -7- (3- morpholine -8- azabicyclo [3.2.1] octane -8- base) -10- oxo -5,10- two The preparation of hydrogen -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 5,5- dimethyl -10- oxo -7- (3- oxo -8- azabicyclo [3.2.1] octane -8- base) -5,10- dihydro - 4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (159mg, 0.38mmol) is dissolved in THF (5mL) and methanol (5mL) In, then it is separately added into acetic acid (0.5mL) and morpholine (67mg, 0.77mmol), sustained response 1 hour under room temperature.Then under ice bath Add sodium cyanoborohydride (73mg, 1.16mmol) in batches, be back to room temperature sustained response 4 hours.Add water quenching to go out reaction, revolve Turn evaporating organic solvent, add EA and water to carry out extraction point liquid, gained organic faciess rotary evaporation removes solvent, inverted system Standby chromatogram purification obtains end-product (40mg, yield 22%).
Molecular formula：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).
Embodiment 65,5- dimethyl -7- (3- methyl -3,8- diazabicyclo [3.2.1] octane -8- base) -10- oxo - The preparation of 5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 6)
(1) tert-butyl group 8- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2- B] indole -7- base) -3,8- diazabicyclo [3.2.1] octane -3- formic acid esters preparation
Will be according to the 2- cyano group -5,5- dimethyl -10- oxo -5,10- two of embodiment 2 (1st) one step preparation method preparation Hydrogen -4H- benzo [f] thieno [3,2-b] indole -7- base triflate (250mg, 0.57mmol) and tert-butyl group 3,8- bis- Azabicyclo [3.2.1] octane -3- formic acid esters (483mg, 2.28mmol) is added in N-Methyl pyrrolidone (20mL), oil bath Lower 120 DEG C are reacted 24 hours.Reaction finishes pours this reactant liquor in water (30mL) into, separates out solid and carries out the sucking filtration that reduces pressure, institute Obtain crude product through column chromatography (DCM:MeOH=40:1) purification obtains product (80mg, yield 28%).
(2) 7- (3,8- diazabicyclo [3.2.1] octane -8- base) -5,5- dimethyl -10- oxo -5,10- dihydro - The preparation of 4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By tert-butyl group 8- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2- B] indole -7- base) -3,8- diazabicyclo [3.2.1] octane -3- formic acid esters (80mg, 0.16mmol) is dissolved in DCM (10mL) In, add TFA (5mL), sustained response 2 hours under room temperature.Then reactant liquor rotary evaporation is removed solvent, add saturation Sodium bicarbonate aqueous solution adjusts pH value to 9, adds DCM and water to carry out extraction point liquid, gained organic faciess rotary evaporation removes solvent Obtain product (60mg, yield 93%).
(3) 5,5- dimethyl -7- (3- methyl -3,8- diazabicyclo [3.2.1] octane -8- base) -10- oxo -5,10- The preparation of dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 7- (3,8- diazabicyclo [3.2.1] octane -8- base) -5,5- dimethyl -10- oxo -5,10- dihydro -4H- Benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (60mg, 0.15mmol) is dissolved in acetonitrile (10mL), adds formalin Solution (61mg, 0.75mmol), sustained response 1 hour under room temperature.Then add sodium cyanoborohydride under ice bath in batches (19mg, 0.3mmol), is back to room temperature sustained response 4 hours.Water quenching is added to go out reaction, rotary evaporation removes organic solvent, addition EA and water carry out extraction point liquid, and gained organic faciess rotary evaporation removes solvent, and residue inverted preparative hplc purification obtains whole product Thing (12mg, yield 19%).
Molecular formula：C24H24N4OS molecular weight：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).
Embodiment 75,5- dimethyl -7- (8- methyl -3,8- diazabicyclo [3.2.1] octane -3- base) -10- oxo - The preparation of 5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 7)
(1) preparation of 8- benzyl 3- tert-butyl group 3,8- diazabicyclo [3.2.1] octane -3,8- dicarboxylic acid esters
By tert-butyl group 3,8- diazabicyclo [3.2.1] octane -3- formic acid esters (820mg, 3.86mmol), N, N- diisopropyl Base ethamine (1.0g, 7.7mmol) is added in dichloromethane (50mL), Deca benzyl chloroformate under ice bath (990mg, 5.8mmol), completion of dropping in 20 minutes, is warmed to room temperature sustained response 4 hours.Reactant liquor adds water and is quenched, ethyl acetate extraction point Liquid, organic faciess rotary evaporation removes solvent and obtains product (1.09g, yield 81%).
(2) preparation of benzyl 3,8- diazabicyclo [3.2.1] octane -8- formic acid esters
By 8- benzyl 3- tert-butyl group 3,8- diazabicyclo [3.2.1] octane -3,8- dicarboxylic acid esters (1.09g, 3.15mmol) it is dissolved in dichloromethane (20mL), add TFA (10mL), sustained response 2 hours under room temperature.Then by body It is that rotary evaporation removes solvent, add saturated sodium bicarbonate aqueous solution and adjust pH value to 9, add dichloromethane to be extracted with water Take a point liquid, gained organic faciess rotary evaporation removes solvent, gained crude product reversed-phase preparative chromatography purification obtains product (715mg, yield 92%).
(3) benzyl 3- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] Indole -7- base) -3,8- diazabicyclo [3.2.1] octane -8- formic acid esters preparation
Will be according to the 2- cyano group -5,5- dimethyl -10- oxo -5,10- two of embodiment 9 (1st) one step preparation method preparation Hydrogen -4H- benzo [f] thieno [3,2-b] indole -7- base triflate (426mg, 0.97mmol), benzyl 3,8- phenodiazine Miscellaneous bicyclic [3.2.1] octane -8- formic acid esters (715mg, 2.9mmol) is dissolved in N-Methyl pyrrolidone (10mL), adds DIPEA (1.25g, 9.7mmol), the lower 120 DEG C of sustained responses of microwave 2 hours.Reaction finishes, and reactant liquor is fallen Enter in water (20mL), separate out solid decompression sucking filtration, gained solid is through column chromatography (DCM:MeOH=50:1) purification obtains product (429mg, yield 82%).
(4) 7- (3,8- diazabicyclo [3.2.1] octane -3- base) -5,5- dimethyl -10- oxo -5,10- dihydro - The preparation of 4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By benzyl 3- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] Indole -7- base) -3,8- diazabicyclo [3.2.1] octane -8- formic acid esters (429mg, 0.80mmol) is added to acetonitrile (10mL) In, slowly Deca Iodotrimethylsilane (0.8g, 4mmol) under room temperature, completion of dropping sustained response 2 hours.Reactant liquor always adds Water (1mL) is quenched reaction, and rotary evaporation removes organic solvent, and residue is through column chromatography (DCM:MeOH=40:1) purification is produced Thing (200mg, yield 62%).
(5) 5,5- dimethyl -7- (8- methyl -3,8- diazabicyclo [3.2.1] octane -3- base) -10- oxo -5,10- The preparation of dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 7- (3,8- diazabicyclo [3.2.1] octane -3- base) -5,5- dimethyl -10- oxo -5,10- dihydro -4H- Benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (200mg, 0.50mmol) is dissolved in acetonitrile (10mL), adds 37% first Aldehyde aqueous solution (202mg, 2.5mmol), sustained response 1 hour under room temperature.Then add sodium cyanoborohydride under ice bath in batches (63mg, 1mmol), is back to room temperature sustained response 4 hours.Water quenching is added to go out reaction, rotary evaporation removes organic solvent, addition second Acetoacetic ester and water carry out extraction point liquid, the rotated evaporation of solvent of organic faciess, and residue is through column chromatography (DCM：MeOH=20:1) Purification obtains end-product (80mg, yield 39%).
Molecular formula：C24H24N4OS molecular weight：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).
Embodiment 85,5- dimethyl -7- (7- methyl -2,7- diaza spiro [3.5] nonane -2- base) -10- oxo -5,10- The preparation of dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 8)
(1) tert-butyl group 2- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2- B] indole -7- base) -2,7- diaza spiro [3.5] nonane -7- formic acid esters preparation
Will be according to the 2- cyano group -5,5- dimethyl -10- oxo -5,10- two of embodiment 13 (1st) one step preparation method preparation Hydrogen -4H- benzo [f] thieno [3,2-b] indole -7- base triflate (100mg, 0.23mmol) and tert-butyl group 2,7- bis- Azaspiro [3.5] nonane -7- formic acid esters (520mg, 2.3mmol) is added in NMP (5mL), 120 DEG C of oil baths 3 hours, adds second Acetoacetic ester (50mL), is washed with water, saturated sodium-chloride water solution, anhydrous sodium sulfate drying, successively through silica gel column chromatography (PE:EA= 1:1) obtain product (100mg, yield 85.%).
(2) 5,5- dimethyl -10- oxo -7- (2,7- diaza spiro [3.5] nonane -2- base) -5,10- dihydro -4H- benzene And the preparation of [f] thieno [3,2-b] indole -2- formonitrile HCN
By tert-butyl group 2- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2- B] indole -7- base) -2,7- diaza spiro [3.5] nonane -7- formic acid esters (100mg, 0.19mmol) is dissolved in DCM (4mL), adds Trifluoroacetic acid (4mL), after being stirred at room temperature 0.5 hour, rotary evaporation removes solvent, adds ammonia and methanol, and rotary evaporation removes molten Agent, residue is through silica gel column chromatography (dichloromethane：Methanol=15:1) purification obtains product (70mg, yield 89%).
(3) 5,5- dimethyl -7- (7- methyl -2,7- diaza spiro [3.5] nonane -2- base) -10- oxo -5,10- two The preparation of hydrogen -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 5,5- dimethyl -10- oxo -7- (2,7- diaza spiro [3.5] nonane -2- base) -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (70mg, 0.17mmol) is dissolved in methanol (3mL), oxolane (3mL) and acetic acid (0.5mL), in mixed solvent, add formaldehyde (73mg), after stirring 1 hour under room temperature, addition sodium cyanoborohydride (53mg, 0.84mmol), it is stirred overnight at room temperature, adds water (50mL), stir out precipitation, filter, filter cake is washed with methanol and obtains end-product (40mg, yield 55%).
Molecular formula：C25H26N4OS molecular weight：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).
Embodiment 95,5- dimethyl -7- (2- methyl -2,7- diaza spiro [3.5] nonane -7- base) -10- oxo -5,10- The preparation of dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 9)
(1) 2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- The preparation of base triflate
By 7- hydroxyl -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- Formonitrile HCN (5.0g, 16.2mmol) is dissolved in pyridine (100mL), be slowly added under ice bath trifluoromethanesulfanhydride anhydride (5.02g, 17.9mmol), it is stirred at room temperature after adding 3 hours, add ethyl acetate extraction, and washed with water, saturated sodium-chloride water solution successively Wash, anhydrous sodium sulfate drying, be spin-dried for obtaining title compound (5.0g, yield 70%).
(2) tert-butyl group 7- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2- B] indole -7- base) -2,7- diaza spiro [3.5] nonane -2- formic acid esters preparation
2- cyano group -5,5- dimethyl -10- oxo-the 5,10- that will prepare by embodiment 12 (1st) one step preparation method Dihydro -4H- benzo [f] thieno [3,2-b] indole -7- base triflate (100mg, 0.23mmol) and the tert-butyl group 2, 7- diaza spiro [3.5] nonane -2- formic acid esters (520mg, 2.3mmol) is added in 5mL NMP, 120 DEG C of oil baths 5 hours, plus Enter 50mL ethyl acetate, use water successively, saturated sodium-chloride water solution washs, anhydrous sodium sulfate drying, silica gel column chromatography（PE：EA= 1:1）Obtain product (110mg, yield 93%).
(3) 5,5- dimethyl -10- oxo -7- (2,7- diaza spiro [3.5] nonane -7- base) -5,10- dihydro -4H- benzene And the preparation of [f] thieno [3,2-b] indole -2- formonitrile HCN
By tertiary fourth 7- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] Indole -7- base) -2,7- diaza spiro [3.5] nonane -2- formic acid esters (100mg, 0.19mmol) is dissolved in DCM (4mL), adds three Fluoroethanoic acid (4mL), after being stirred at room temperature 0.5 hour, rotary evaporation removes solvent, adds ammonia and methanol, and rotary evaporation removes molten Agent, residue silica gel column chromatography (dichloromethane：Methanol=15:1) purification obtains product (79mg, yield 98%).
(4) 5,5- dimethyl -7- (2- methyl -2,7- diaza spiro [3.5] nonane -7- base) -10- oxo -5,10- two The preparation of hydrogen -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 5,5- dimethyl -10- oxo -7- (2,7- diaza spiro [3.5] nonane -7- base) -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (70mg, 0.17mmol) is dissolved in methanol (3mL), oxolane (3mL) and acetic acid (0.5mL) in mixed solvent, add formaldehyde (37% aqueous solution) (73mg, 0.9mmol), stirring under room temperature added cyanogen after 1 hour Base sodium borohydride (53mg, 0.84mmol), is stirred overnight at room temperature, and adds water (50mL), stirs out precipitation, filters, filter cake first Alcohol washs to obtain end-product (53mg, yield 73%).
Molecular formula：C25H26N4OS molecular weight：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).
Embodiment 105,5- dimethyl -7- (6- methyl -2,6- diaza spiroheptane -2- base) -10- oxo -5, The preparation of 10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 10)
(1) tert-butyl group 6- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2- B] indole -7- base) -2,6- diaza spiroheptane -2- formic acid esters preparation
Will be according to the 2- cyano group -5,5- dimethyl -10- oxo -5,10- two of embodiment 9 (1st) one step preparation method preparation Hydrogen -4H- benzo [f] thieno [3,2-b] indole -7- base triflate (100mg, 0.23mmol) and tert-butyl group 2,6- bis- Azepine spiroheptane -2- formic acid esters (455mg, 2.3mmol) is added in NMP (5mL), 120 DEG C of oil baths 3 hours, adds second Acetoacetic ester (50mL), uses water successively, and saturated sodium-chloride water solution washs, anhydrous sodium sulfate drying, silica gel column chromatography (PE：EA=1: 1) obtain product (100mg, yield 90%).
(3) 5,5- dimethyl -10- oxo -7- (2,6- diaza spiroheptane -2- base) -5,10- dihydro -4H- benzene And the preparation of [f] thieno [3,2-b] indole -2- formonitrile HCN
By tert-butyl group 6- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2- B] indole -7- base) -2,6- diaza spiroheptane -2- formic acid esters (100mg, 0.20mmol) is dissolved in DCM (4mL), adds Trifluoroacetic acid (4mL), after being stirred at room temperature 0.5 hour, rotary evaporation removes solvent, adds ammonia and methanol, and rotary evaporation removes molten Agent, residue is through silica gel column chromatography (dichloromethane：Methanol=15:1) purification obtains product (70mg, yield 88%).
(4) 5,5- dimethyl -7- (6- methyl -2,6- diaza spiroheptane -2- base) -10- oxo -5,10- two The preparation of hydrogen -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 5,5- dimethyl -10- oxo -7- (2,6- diaza spiroheptane -2- base) -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (66mg, 0.17mmol) is dissolved in methanol (3mL), oxolane (3mL) and acetic acid (0.5mL) in mixed solvent, add formaldehyde (37% aqueous solution) (73mg, 0.9mmol), stirring under room temperature added cyanogen after 1 hour Base sodium borohydride (53mg, 0.84mmol), is stirred overnight at room temperature, and adds 50mL water, stirs out precipitation, filters, filter cake methanol Washing obtains end-product (30mg, yield 44%).
Molecular formula：C23H22N4OS molecular weight：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).
Embodiment 115,5- dimethyl -7- (9- methyl -3,9- diaza spiro [5.5] hendecane -3- base) -10- oxo -5, The preparation of 10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 11)
(1) tert-butyl group 9- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2- B] indole -7- base) -3,9- diaza spiro [5.5] hendecane -3- formic acid esters preparation
Will be according to the 2- cyano group -5,5- dimethyl -10- oxo -5,10- two of embodiment 9 (1st) one step preparation method preparation Hydrogen -4H- benzo [f] thieno [3,2-b] indole -7- base triflate (100mg, 0.23mmol) and tert-butyl group 3,9- bis- Azaspiro [5.5] hendecane -3- formic acid esters (585mg, 2.3mmol) is added in NMP (5mL), 120 DEG C of oil baths 3 hours, adds Ethyl acetate (50mL), is washed with water, saturated sodium-chloride water solution, anhydrous sodium sulfate drying, silica gel column chromatography (PE successively：EA= 1:1) obtain product (100mg, yield 78%).
(2) 5,5- dimethyl -10- oxo -7- (3,9- diaza spiro [5.5] hendecane -3- base) -5,10- dihydro -4H- The preparation of benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By tert-butyl group 9- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2- B] indole -7- base) -3,9- diaza spiro [5.5] hendecane -3- formic acid esters (100mg, 0.18mmol) is dissolved in DCM (4mL), plus Enter trifluoroacetic acid (4mL), after being stirred at room temperature 0.5 hour, rotary evaporation removes solvent, add ammonia and methanol, rotary evaporation removes Solvent, residue silica gel column chromatography (dichloromethane：Methanol=15:1) purification obtains product (80mg, yield 98%).
(3) 5,5- dimethyl -7- (9- methyl -3,9- diaza spiro [5.5] hendecane -3- base) -10- oxo -5,10- two The preparation of hydrogen -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 5,5- dimethyl -10- oxo -7- (3,9- diaza spiro [5.5] hendecane -3- base) -5,10- dihydro -4H- benzene And [f] thieno [3,2-b] indole -2- formonitrile HCN (75mg, 0.17mmol) is dissolved in methanol (3mL), oxolane (3mL) and acetic acid (0.5mL) in mixed solvent, add formaldehyde (73mg, 0.85mmol), after stirring 1 hour under room temperature, add cyano group hydroboration Sodium (53mg, 0.84mmol), is stirred overnight at room temperature, and adds water (50mL), stirs out precipitation, filters, and filter cake is washed with methanol End-product (50mg, yield 64%).
Molecular formula：C27H30N4OS molecular weight：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).
Embodiment 125,5- dimethyl -7- (cis -5- methyl hexahydropyrrolo simultaneously [3,4-c] pyrroles -2- (1H)-yl) -10- The preparation of oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 13)
(1) 2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- The preparation of base triflate
Will according to embodiment 1 (1st) step the-the (8) one step preparation method preparation 7- hydroxyl -5,5- dimethyl -10- oxo - 5,10- dihydros -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (5.0g, 16.2mmol) are dissolved in pyridine (100mL), It is slowly added to trifluoromethanesulfanhydride anhydride (5.02g, 17.8mmol) under ice bath, is stirred at room temperature after adding 3 hours, add ethyl acetate Extraction, and washed with water, saturated sodium-chloride water solution successively, anhydrous sodium sulfate drying, rotary evaporation removes solvent and obtains product (5.2g, yield 73%).
(2) cis -5- of the tert-butyl group (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- base) hexahydropyrrolo simultaneously [3,4-c] pyrroles -2 (1H)-formic acid esters preparation
By 2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- Base triflate (100mg, 0.23mmol) and the tert-butyl group cis-hexahydropyrrolo simultaneously [3,4-c] pyrroles -2 (1H)-formic acid esters (488mg, 2.3mmol) is added in NMP (5mL), 120 DEG C of oil baths 3 hours, adds ethyl acetate (50mL), uses water, full successively With sodium-chloride water solution washing, anhydrous sodium sulfate drying, silica gel column chromatography (PE:EA=1:1) obtain product (100mg, yield 87%).
(3) 7- (cis-hexahydropyrrolo simultaneously [3,4-c] pyrroles -2 (1H)-yl) -5,5- dimethyl -10- oxo -5,10- two The preparation of hydrogen -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By cis for the tert-butyl group -5- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- base) simultaneously [3,4-c] pyrroles -2 (1H)-formic acid esters (100mg, 0.20mmol) are dissolved in DCM to hexahydropyrrolo (4mL), add trifluoroacetic acid (4mL), after being stirred at room temperature 0.5 hour, rotary evaporation removes solvent, add ammonia and methanol, rotation Evaporation of solvent, residue silica gel column chromatography (dichloromethane：Methanol=15:1) purification obtains product (78mg, yield 97%).
(4) 5,5- dimethyl -7- (cis -5- methyl hexahydropyrrolo simultaneously [3,4-c] pyrroles -2 (1H)-yl) -10- oxo - The preparation of 5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By 7 (cis-hexahydropyrrolo simultaneously [3,4-c] pyrroles -2 (1H)-yl) -5,5- dimethyl -10- oxo -5,10- two Hydrogen -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (68mg, 0.17mmol) is dissolved in methanol (3mL), oxolane (3mL) and in the mixed solvent of acetic acid (0.5mL), add formaldehyde (73mg, 0.89mmol), stirring under room temperature added after 1 hour Sodium cyanoborohydride (53mg, 0.84mmol), is stirred overnight at room temperature, and adds water (50mL), stirs out precipitation, filters, and filter cake is used Methanol washs to obtain end-product (40mg, yield 57%).
Molecular formula：C24H24N4OS molecular weight：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).
Embodiment 137- (external form -6- amino-3-azabicyclo [3.1.0] hexane -3- base) -5,5- dimethyl -10- oxygen The preparation in generation -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN (compound 15)
(1) tert-butyl group external form -6- (((benzyloxy) carbonyl) amino) -3- azabicyclo [3.1.0] hexane -3- formic acid esters Preparation
By tert-butyl group external form -6- amino-3-azabicyclo [3.1.0] hexane -3- formic acid esters (4.15g, 21mmol), N, N- Diisopropylethylamine (5.4g, 42mmol) is added in dichloromethane (100mL), Deca benzyl chloroformate under ice bath (5.36g, 31mmol), completion of dropping in 20 minutes, is warmed to room temperature, sustained response 4 hours.Add water (100mL) to carry out extraction point liquid, have Machine phase rotary evaporation removes solvent and obtains product (5.0g, yield 72%).
(2) preparation of benzyl (external form -3- azabicyclo [3.1.0] hexane -6- base) carbamate
By tert-butyl group external form -6- (((benzyloxy) carbonyl) amino) -3- azabicyclo [3.1.0] hexane -3- formic acid esters (5.0g, 15.0mmol) is dissolved in dichloromethane (20mL), adds TFA (10mL), sustained response 2 hours under room temperature.So Rotary evaporation removes solvent afterwards, adds saturated sodium bicarbonate aqueous solution and adjusts pH value to 9, adds DCM and water to carry out extraction point Liquid, organic faciess rotary evaporation is removed solvent, and gained crude product reversed-phase preparative chromatography purification obtains product (3.0g, yield 86%).
(3) benzyl (external form -3- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- base) -3- azabicyclo [3.1.0] hexane -6- base) carbamate preparation
Will be according to the 2- cyano group -5,5- dimethyl -10- oxo -5,10- two of embodiment 2 (1st) one step preparation method preparation Hydrogen -4H- benzo [f] thieno [3,2-b] indole -7- base triflate (360mg, 0.82mmol), benzyl (external form -3- Azabicyclo [3.1.0] hexane -6- base) carbamate (569mg, 2.45mmol) is dissolved in N-Methyl pyrrolidone (10mL) In, add DIPEA (1.06g, 8.2mmol), the lower 120 DEG C of sustained responses of microwave 2 hours.Reaction finishes, will Reactant liquor is poured into water, and separates out solid decompression sucking filtration, gained solid is through column chromatography (DCM:MeOH=50:1) purification obtains product (134mg, yield 31%).
(4) 7- (external form -6- amino-3-azabicyclo [3.1.0] hexane -3- base) -5,5- dimethyl -10- oxo -5, The preparation of 10- dihydro -4H- benzo [f] thieno [3,2-b] indole -2- formonitrile HCN
By benzyl (external form -3- (2- cyano group -5,5- dimethyl -10- oxo -5,10- dihydro -4H- benzo [f] thieno [3,2-b] indole -7- base) -3- azabicyclo [3.1.0] hexane -6- base) carbamate (134mg, 0.26mmol) is added to In acetonitrile (10mL), slowly Deca Iodotrimethylsilane (260mg, 1.3mmol) under room temperature, completion of dropping sustained response 2 hours. Water quenching is added to go out reaction in reactant liquor, rotary evaporation removing solvent, residue is through column chromatography (DCM:MeOH=40:1) purification obtains end Product (40mg, 40%).
Molecular formula：C22H20N4OS molecular weight：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 (10)

1. lead to the compound shown in formula II or its stereoisomer or its pharmaceutically acceptable salt：
Wherein,
R1、R2And R3It is respectively selected from hydrogen or C1-4Alkyl；
Q be selected from 5～6 circle heterocycles bases, 8 yuan of bridge heterocyclic radicals, 7～11 yuan of spiro heterocyclic radicals or
R4Selected from following groups：
(1) hydrogen, amino or C1-4Alkyl,
(2) 8 yuan of bridge heterocyclic radicals or 5～6 circle heterocycles bases；
N is selected from 0 or 1, when n is 0, R4Do not exist,
And when Q is selected from 5～6 circle heterocycles base, R45～6 circle heterocycles bases can not be selected from；
R5And R6It is respectively selected from hydrogen or C1-4Alkyl；
R7Selected from hydrogen, cyano group, hydroxyl, amino, methyl, ethyl or chlorine atom；
M is selected from N-R8, R8Selected from hydrogen or C1-4Alkyl；
Y is selected from C-R9, R9Selected from hydrogen, methyl, ethyl or n-pro-pyl；
X is selected from S.
2. compound as claimed in claim 1 or its stereoisomer or its pharmaceutically acceptable salt：Wherein,
Q is selected from 5～6 circle heterocycles bases；
R4Selected from 8 yuan of bridge heterocyclic radicals；
N is selected from 1.
3. compound as claimed in claim 1 or its stereoisomer or its pharmaceutically acceptable salt：Wherein,
Q is selected from 8 yuan of bridge heterocyclic radicals；
R4Selected from hydrogen, amino, C1-4Alkyl or 5～6 circle heterocycles bases；
N is selected from 0 or 1, when n is 0, R4Do not exist.
4. compound as claimed in claim 1 or its stereoisomer or its pharmaceutically acceptable salt：
Wherein,
Q be selected from 7～11 yuan of spiro heterocyclic radicals or
R4Selected from hydrogen, amino or C1-4Alkyl；
N is selected from 0 or 1, when n is 0, R4Do not exist.
5. compound as claimed in claim 1, its pharmaceutically acceptable salt, its stereoisomer, described compound is selected from：
6. compound as described below, its pharmaceutically acceptable salt, its stereoisomer, described compound is selected from：
7. a kind of pharmaceutical composition, it include compound described in claim 1-6 any claim or its stereoisomer, Or its pharmaceutically acceptable salt and one or more pharmaceutical carrier and/or diluent.
8. pharmaceutical composition as claimed in claim 7 is it is characterised in that also can contain one or more antitumor agent and immunity Inhibitor, described antitumor agent and immunosuppressant are selected from methotrexate, Capecitabine, gemcitabine, Doxifluridine, training U.S. bent plug disodium, pazopanib, imatinib, erlotinib, Lapatinib, gefitinib, ZD6474, Herceptin, shellfish are cut down Monoclonal antibody, Rituximab, Herceptin, paclitaxel, vinorelbine, docetaxel, doxorubicin, hydroxy camptothecin, mitogen Mycin, epirubicin, Pirarubicin, bleomycin, letrozole, tamoxifen, fulvestrant, music score of Chinese operas Rayleigh, flutamide, bright third Rayleigh, Anastrozole, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, chlormethine, Ma Fa Orchid, Chlorambucil, carboplatin, cisplatin, oxaliplatin, network platinum, Topotecan, camptothecine, topotecan, everolimuses, Sirolimus, Special cancer fit, Ismipur, 6- thioguanine, azathioprine, rhzomorph D, daunorubicin, amycin, mitoxantrone, win honour for mould Element, plicamycin or aminoglutethimide.
9. the compound as described in claim 1-6 any claim or its stereoisomer or it is pharmaceutically acceptable Salt preparation for treat and/or prevent ALK mediation cancer-related diseases medicine in application, described cancer is related Disease be selected from cerebroma, lung cancer in non-cellule type, squamous cell cancer, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, cancer of pancreas, Breast carcinoma, head and neck cancer, cervical cancer, carcinoma of endometrium, rectal cancer, hepatocarcinoma, renal carcinoma, adenocarcinoma of esophagus, esophageal squamous cell carcinoma, god Through glioma, carcinoma of prostate, female reproductive tract cancer, lymphoma, neuroblastoma, neurofibromatosiss, osteocarcinoma, skin carcinoma, Colon cancer, carcinoma of testis, small cell lung cancer, gastrointestinal stromal tumor, mast cell tumor.
10. the compound as described in claim 1-6 any claim or its stereoisomer or it is pharmaceutically acceptable Salt preparation for treat and/or prevent ALK mediation cancer-related diseases medicine in application, described cancer is related Disease is selected from incidence squamous cytoma, thyroid carcinoma, hepatoblastoma, papillary renal cell tumor, nephroblastoma, prostate Tumor, multiple myeloma or melanoma.
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CN1195349A (en) * 1995-07-14 1998-10-07 艾科斯有限公司 Cyclic GMP-specific phosphodiesterase inhibitors
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
CN102177134A (en) * 2008-08-08 2011-09-07 麻州四期制药公司 C7-fluoro substituted tetracycline compounds
CN102459172A (en) * 2009-06-10 2012-05-16 中外制药株式会社 Tetracyclic compound

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Publication number Priority date Publication date Assignee Title
CN1195349A (en) * 1995-07-14 1998-10-07 艾科斯有限公司 Cyclic GMP-specific phosphodiesterase inhibitors
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
CN102177134A (en) * 2008-08-08 2011-09-07 麻州四期制药公司 C7-fluoro substituted tetracycline compounds
CN102459172A (en) * 2009-06-10 2012-05-16 中外制药株式会社 Tetracyclic compound

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