CN105732638B

CN105732638B - A kind of bruton's tyrosine kinase inhibitor with loop coil or caged scaffold and preparation method thereof

Info

Publication number: CN105732638B
Application number: CN201610046120.4A
Authority: CN
Inventors: 李英富; 黄浩喜; 刘冠峰; 陈垌辉; 杜振军
Original assignee: CHENGDU BRILLIANT PHARMACEUTICAL Co Ltd
Current assignee: Chengdu Beite Pharmaceutical Co., Ltd
Priority date: 2016-01-22
Filing date: 2016-01-22
Publication date: 2018-01-30
Anticipated expiration: 2036-01-22
Also published as: CN105732638A; WO2017124657A1

Abstract

The invention discloses a kind of bruton's tyrosine kinase inhibitor with loop coil or caged scaffold and preparation method thereof, the inhibitor includes formula (I), the inhibitor has obvious inhibitory action to BTK activity, on a cellular level, nearly 8 times are improved to the inhibitory activity of DOHH2 cells, loop coil or endocyclic compound provided by the invention can be used as BTK inhibitor, have the application prospect of wide anti-malignant tumor.

Description

A kind of bruton's tyrosine kinase inhibitor and its system with loop coil or caged scaffold Preparation Method

Technical field

The invention belongs to medicinal chemistry art, and in particular to a kind of bruton's tyrosine with loop coil or caged scaffold swashs Enzyme inhibitor, and the preparation method of such compound and the method and purposes that suppress BTK activity using these new compounds.

Background technology

In recent years, the treatment method of Malignancy is just progressively from the chemoimmunotherapy of routine to targeted therapy Transformation, different from classic chemotherapy medicine, targeted therapy can be accurately positioned the cancer cell of fast-growth, while and can avoids aligning The injury of normal cell, reduces the toxic side effects such as alopecia, gastrointestinal reaction and the bone marrow suppression of classic chemotherapy medicine；On the other hand with Afatinib, Cl 1033, HKI-272 etc., covalently anti-tumor drugs targeting particularly largely replaces Buddhist nun's class medicine for kinases The generation of thing, people are made to increasingly focus on small molecule covalency inhibitor medicine again.Small molecule covalency inhibitor (covalent Inhibitors irreversible inhibitor (irreversible inhibitours)) is also referred to as, is residual with target protein by covalent bond Irreversible binding occurs for base, so as to play a kind of inhibitor of its biological function.

B cell malignant tumour includes chronic lymphocytic leukemia (CLL), ALL (ALL), small leaching Bar cell lymphoma (SLL), lymphoma mantle cell (MCL), follicular lymphoma (FL), Diffuse Large B-Cell Lymphoma (DLBCL), Huppert's disease (MM) andMacroglobulinemia (WM).For above-mentioned illness conventional medicament Although the survival of patients time can not can effectively be extended with relief of symptoms.

Bruton's tyrosine kinase (BTK) is a kind of member of non-receptor protein tyrosine kinase family, is except T lymphs The key signal enzyme expressed in all hematopoetic cell types outside cell and NK.On B cell signal transduction way Play the part of vital role in footpath, it is closely related with bone-marrow-derived lymphocyte development, differentiation, signal transmission and survival.BTK is in B cell The important function of acceptor (BCR) signal path, become the popular target spot of B cell treating malignant tumor.

The current first oral BTK inhibitor for obtaining FDA approval listings replaces Buddhist nun according to Shandong, granted four kinds of indications, Its inhibitor is selectively strong, and toxic side effect is low, is referred to as " breakthrough " new drug, its main mechanism be with the Guang of BTK enzymes half by The sulfydryl of sour (Cys481) residue is had an effect, and is formed covalent bond, BTK enzymes is lost activity.But in administration process, still deposit Easily it is being metabolized, bioavilability is low (only 7~23%), the problems such as clinical administration amount big (560mg/ days).

The content of the invention

A kind of bruton's tyrosine with loop coil or caged scaffold that the present invention provides for above-mentioned weak point swashs Enzyme inhibitor, particularly formula (I), (II) compound and its optical isomer or its salt pharmaceutically received, hydrate or molten Agent compound individually or with other drugs is combining the purposes as BTK inhibitor in medicine, is specifically preventing or is controlling The purposes in cell proliferative diseases such as cancer is treated, it is especially thin in prevention or treatment lymphoma mantle cell (MCL) and chronic lymphatic Purposes in the B cell malignant tumor medicines such as born of the same parents' leukaemia (CLL).

A kind of bruton's tyrosine kinase inhibitor of loop coil or caged scaffold, including formula (I)：

Wherein Y is selected from following groups：

Wherein, R1, R2 can be independently selected from hydrogen atom, fluorine atom, bromine atoms, iodine atom, amino, cyano group, sulfydryl, C₁ ~C₅Straight or branched alkyl；One or more hydrogen atoms on the alkyl can be by one or more halogen, amino, cyanogen Base, hydroxyl, sulfenyl substitution；

m₁、m₂It is C₁~C₅Alkyl, one or more carbon atom in the alkyl can by one or more O, S ,-C=O ,-C=S ,-S=O ,-S (=O)₂Substitution；

n₁、n₂It is C₀~C₅Alkyl, one or more carbon atom in the alkyl can by one or more O, S ,-C=O ,-C=S ,-S=O ,-S (=O)₂Substitution；

G is C, O, S ,-S=O ,-S (=O)₂Or NR⁴；The R⁴Can be hydrogen, C₁~C₆Straight or branched alkyl, C₁~C₆ Straight or branched miscellaneous alkyl；Wherein C₁~C₆Straight or branched alkyl, C₁~C₆One or more hydrogen in straight or branched miscellaneous alkyl Atom further can be substituted by one or more amino, cyano group, hydroxyl, sulfenyl, halogen.

More preferably contain the compound of formula (II)：

Wherein, m₁、m₂It is C₁~C₂Alkyl, one or more carbon atom in the alkyl can be by one or more Individual O, S ,-C=O ,-C=S, S=O, S (=O)₂Substitution；

n₁、n₂It is C₁~C₃Alkyl, one or more carbon atom in the alkyl can by one or more O, S ,-C=O ,-C=S, S=O, S (=O)₂Substitution；

G is C, O, S, S=O, S (=O)₂Or NR⁴；The R⁴Can be hydrogen, C₁~C₄Straight or branched alkyl, C₁~C₄Directly Chain or branched heteroalkyl groups.

Further, the bruton's tyrosine kinase inhibitor of loop coil or caged scaffold, selected from following structure：

Further, a kind of bruton's tyrosine kinase inhibitor with loop coil or caged scaffold, selected from following structure：

Provided by the invention a kind of have loop coil or the bruton's tyrosine kinase inhibitor of caged scaffold and its preparation side Method, there are following several beneficial effects：

(1) bruton's tyrosine kinase inhibitor provided by the invention is lived to BTK and cell propagation with stronger suppression Property, it is higher nearly 8 times for Buddhist nun's drug effect than according to Shandong, therefore use can reach prevention and treatment effect compared with small pit goaf, utilization rate is high.

(2) loop coil designed by the present invention or endocyclic compound can be used as BTK inhibitor and have wide anti-malignant tumor Application prospect, can such as treat chronic lymphocytic leukemia, chronic lymphocytic leukemia (CLL), B cell young lamphocyte Leukaemia, lymph chylema cell lymphoma, splenic Marginal Zone Lymphoma, ALL (ALL), small lymphocyte Property lymthoma (SLL) plasma cell myeloma, plasmacytoma, knot outward flange band B cell lymphoma, Huppert's disease (MM), It is the chronic lymphocytic leukemia of chromosome deficiency, knot inward flange band B cell lymphoma, lymphoma mantle cell (MCL), intravascular Large B cell lymphoid tumor, lymphoma primary effusion, diffusivity B cell lymphoma (DLBCL), follicular lymphoma, WaldenstromShi macroglobulinemias etc.；It can be additionally used in systemic loupus erythematosus, rheumatoid arthritis, systemic vascular Inflammation, chorionitis, pemphigus, dermatomyositis, MCTD, autoimmune hemolytic anemia, thyroid autoimmune It is disease, ulcerative colitis, vaginitis, allergic rhinitis, bronchitis, mastitis, fibrositis, cervicitis, oaritis, preceding Row adenositis, rhinitis, nasosinusitis, tonsillitis, pericarditis, pneumonia, ephritis, enteritis, appendicitis, cholecystitis, encephalitis, conjunctivitis, The treatment of endometritis, meningitis, pharyngitis etc..

Embodiment

Following abbreviation has meaning as follows：DMSO represents dimethyl sulfoxide；DMF represents N, N- dimethyl formyls Amine；LDA represents lithium diisopropylamine；LHDMS represents two (trimethyl silicon substrate) lithium amides；LAH represents lithium aluminium hydride reduction；DCM tables Show dichloromethane；TEA represents triethylamine；TFA represents trifluoroacetic acid；THF represents tetrahydrofuran；PPh₃Represent triphenylphosphine；EA Represent ethyl acetate；MeOH represents methanol；DIEA represents N, N diisopropylethylamine, (BOC)₂O represents di-tert-butyl dicarbonate； OTs is to represent p-methyl benzenesulfonic acid ester；OMs is to represent methanesulfonate ester；9-BBN represents bicyclic [3.3.1] nonane of 9- boron；Rt tables Show room temperature；DIAD represents diisopropyl azodiformate.

Derivative shown in the formula (I) of the present invention can be prepared by formulas below：

Initiation material passes through for commercialization according to Shandong for Buddhist nun's intermediate A and the segments such as the loop coil with protection group or bridged ring Mitsunobu reacts or directly substitution reaction obtains intermediate B, and intermediate B generates intermediate C by deprotection reaction, raw Into C further act on obtaining target compound with acryloyl chloride.The structure of compound is determined by HNMR and LC-MS.

HNMR measure is to use Bruker 400 (400MHz) spectrometer,¹H chemical shifts are provided with ppm units, are surveyed It is deuterochloroform or deuterated DMSO to determine solvent, is inside designated as tetramethylsilane, chemical shift provides using ppm as unit.

LC-MS measure be with the series of Agilent 1200,6110 series, 6120 series, using electrospray ionization pattern, Condition is：Waters X Bridge C18 posts (50mm x 4.6mm x 3.5um), flow velocity：2.0mL/min, column temperature：40℃.

Thin layer silica gel uses GF254 silica gel plates.

Column chromatography is carrier using Yantai Huanghai Sea silica gel 200-300 mesh silica gel.

Embodiment 1：1- (3- (amino -3- (4- Phenoxyphenyls) -3a, 7a-1H- pyrazoles [3,4-d] pyrimidine -1- bases) -2, 3- dihydros spiral shell [indenes-Isosorbide-5-Nitrae '-piperidines] -1'- bases) propyl- 2- alkene -1- ketone preparation,

Synthesis step is as follows：

Step 1:The preparation of double (2- chloroethyls) t-butyl carbamates (1b) of N, N-

Under condition of ice bath, to equipped with two (2- chloroethyls) amine hydrochlorates (19.99g, 0.112mmol) and (Boc)₂O TEA (34.00g, 0.336mmol), question response mixture are slowly added into the 300mL DCM solution of (26.84g, 0.123mmol) Naturally it is increased to room temperature and continues to stir 5h, after TLC shows that raw material reaction is completed, reaction solution uses water and saturated aqueous common salt respectively Washing, organic phase anhydrous Na₂SO₄Fully dry, pass through flash chromatography (PE after vacuum evaporation：EA=100：1) purifying obtains 24.4g target compounds, it is colorless oil；

Step 2:The preparation of spiral shell [indenes -1,4- piperidines] -1- carboxylic acid tert-butyl esters (1d)

Under nitrogen protection, slowly dripped at -10 DEG C into the 50mL THF solutions equipped with indenes (2.50g, 21.52mmol) Add LiTMDS (43.04mL, 43.04mmol, 1mol/L THF), after the reactant mixture stirs 30min at -10 DEG C, - N is added dropwise at 10 DEG C into above-mentioned reactant mixture, double (2- chloroethyls) t-butyl carbamates (5.21g, 21.52mmol) of N- 5mL THF solutions, the reactant mixture stir 2h at -10 DEG C, and 30min is then stirred at room temperature, and TLC display reactions are completed Afterwards, water adds in reactant mixture and then extracts (50mL X 3), organic phase anhydrous Na with EA₂SO₄Fully dry, vacuum is steamed Pass through flash chromatography (PE after hair：EA=50：1~30：1) purifying obtains 3.6g target compounds, is yellow solid；

Step 3:The preparation of 3- hydroxyl -2,3- dihydros spiral shell [indenes -1,4'- piperidines] -1'- carboxylic acid tert-butyl esters (1e)

Under nitrogen protection, to the 30mL equipped with spiral shell [indenes-Isosorbide-5-Nitrae-piperidines] -1- carboxylic acid tert-butyl esters (2.85g, 0.01mmol) 9-BBN (40.00mL, 0.02mol, 0.5mol/L THF) is slowly added dropwise in (0 DEG C) in THF solution；The reactant mixture is at 70 DEG C Under be stirred overnight, state 1mol/L NaOH (20.00mL) and H are slowly added dropwise in reactant mixture then up₂O₂(4.00mL) quenches Go out reaction, be subsequently cooled to extract (50mL X 3), organic phase anhydrous Na with EA after room temperature₂SO₄Fully dry, be evaporated in vacuo Pass through chromatography (PE afterwards：EA=5：1) purifying obtains 2.85g target compounds, is yellow oil；

1H NMR(400MHz,CDCl₃) δ 1.39 (1H, dd, J=13.2Hz, 2.0Hz), 1.49 (9H, s), 1.63 (1H, Dd, J=13.2Hz, 2.4Hz), 1.71-1.78 (1H, m), 1.90-1.98 (2H, m), 2.52 (1H, dd, J=13.6Hz, 7.2Hz),2.88-2.96(2H,m),5.26-5.29(1H,m),7.18-7.20(1H,m),7.28-7.35(2H,m),7.41- 7.43(1H,m).

Step 4:3- (4- amino -3- (Phenoxyphenyl) -1H pyrazoles [3,4-d] pyrimidine -1- bases) -2,3- dihydros spiral shell [indenes - 1,4'- piperidines] -1'- bases) propyl- 2- alkene -1- t-butyl formates (1f) preparation

Under nitrogen protection, to equipped with 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (B-H) (304mg, 1.00mmol), 3- hydroxyl -2,3- dihydros spiral shell [indenes-Isosorbide-5-Nitrae '-piperidines] -1'- carboxylic acid tert-butyl esters (456mg, 1.50mmol) and PPh₃In the 20mL THF solutions of (515mg, 1.50mmol) (0 DEG C) slowly be added dropwise DIAD (303mg, 1.50mmol), the reactant mixture, which is stirred at room temperature after 3h, is slowly added dropwise water quenching and goes out reaction, EA extractions (50mL x 3), has Machine mutually uses anhydrous Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=60：1~10:1) purifying obtains 280mg target compounds, are white solid, yield 47.6%.

Step 5:1- (2,3- dihydros spiral shell [indenes -1,4'- piperidines] -3- bases) -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4- D] pyrimidine -4- amine (1g) preparation；

At room temperature, to equipped with 3- (4- amino -3- (Phenoxyphenyl) -1H pyrazoles [3,4-d] pyrimidine -1- bases) -2,3- Dihydro spiral shell [indenes-Isosorbide-5-Nitrae '-piperidines] -1'- bases) propyl- 2- alkene -1- t-butyl formates (500mg, 0.85mmol) 20mL DCM solution In TFA (2mL) is slowly added dropwise, the reactant mixture is stirred at room temperature 2h, NaHCO is then slowly added dropwise₃(1mol/L) is to pH More than 7, organic phase is separated, aqueous phase extracts (20mL x 3) with DCM, merges above-mentioned organic phase anhydrous Na₂SO₄Fully dry, very Pass through chromatography (DCM after sky evaporation：MeOH=10：1~1:1) purifying obtains 320mg target compounds, is white solid, production Rate is 77.1%；

Step 6:1- (3- (amino -3- (4- Phenoxyphenyls) -3a, 7a-1H- pyrazoles [3,4-d] pyrimidine -1- bases) -2,3- Dihydro spiral shell [indenes -1,4'- piperidines] -1'- bases) propyl- 2- alkene -1- ketone (1) preparation

Under nitrogen protection, to equipped with 1- (2,3- dihydro spiral shells [indenes-Isosorbide-5-Nitrae '-piperidines] -3- bases) -3- (4- phenoxy group benzene Base) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (320mg, 0.65mmol), TEA (132mg, 1.31mmol) 20mL DCM solution In (0 DEG C) acryloyl chloride (56mg, 0.62mmol) is slowly added dropwise, the reactant mixture is stirred at room temperature 3h, then slowly drips Add saturation NH₄Reaction is quenched in Cl, is washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄Fully dry, lead to after vacuum evaporation Cross chromatography (DCM：MeOH=20：1) purifying obtains 220mg target compounds, is white solid, yield 62.3%；

1H NMR(400MHz,d₆-DMSO)δ1.64-1.70(2H,m),1.89-2.02(2H,m),2.55-2.58(2H, m),2.88-2.93(2H,m),3.20-3.26(0.5H,m),3.45-3.51(0.5H,m),4.10-4.15(1H,m),4.50- 4.54 (1H, m), 5.69 (1H, dd, J=10.4Hz, 2.4Hz), 6.14 (1H, dd, J=16.4Hz, 2.4Hz), 6.55 (1H, t, ), J=7.6Hz 6.85-6.92 (2H, m), 7.08-7.19 (6H, m), 7.29 (1H, t, J=7.6Hz), 7.36-7.43 (3H, M), 7.59 (2H, d, J=8.8Hz), 8.32 (1H, s)

EM (calculated value)：542.2；MS(ESI)m/e(M+1H)⁺：543.3

Embodiment 2

Inner mold -1- ((1R, 3s, 5S) -3- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- Base) -8- azepines bridge [3.2.1] octane -8- bases) propyl- 2- alkene -1- ketone and external form -1- ((1R, 3s, 5S) -3- (4- amino -3- (4- Phenoxyphenyl) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -8- azepines bridge [3.2.1] octane -8- bases) propyl- 2- alkene -1- ketone Prepare

Step 1:The preparation of N- tertbutyloxycarbonyls-notropinon

Under nitrogen protection, to equipped with notropinon hydrochloride (4.8g, 29.70mmol) and TEA (4.50g, It is slowly added into (Boc) in DCM (150mL) solution 44.55mmol)₂O (56mg, 44.55mmol), the reactant mixture is in room It is stirred overnight under temperature, adds water (50mL) after the completion of reaction again and reaction is quenched, organic phase is separated, aqueous phase extracts (30mL with DCM X 3), merge above-mentioned organic phase anhydrous Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=5：1~1:1) Purifying obtains 5.0g target compounds, is white solid, yield 74.6%；

Step 2:3- external forms-hydroxyl -8- azepines bridge [3.2.1] octane -8- t-butyl formates, 3- inner mold-hydroxyl -8- azepines It is prepared by bridge [3.2.1] octane -8- t-butyl formates

At room temperature, to the 25mL THF equipped with N- tertbutyloxycarbonyls-notropinon (2.25g, 10.00mmol) and NaBH is added portionwise in 10mL MeOH mixed solution₄(756mg, 20.00mmol), the reactant mixture are stirred at room temperature 1h, saturation NH is slowly added dropwise after the completion of reaction again₄Reaction is quenched in Cl, and (40mL x 3), anhydrous Na are extracted with EA₂SO₄It is fully dry It is dry, pass through chromatography (PE after vacuum evaporation：EA=4：1~3:1) purifying obtains 722mg target compounds 3- external forms-hydroxyl -8- Azepine bridge [3.2.1] octane -8- t-butyl formates, yield 31.8%；It is pungent to obtain 3- inner mold-hydroxyl -8- azepines bridge [3.2.1] Alkane -8- t-butyl formate 1.48g, yield 65.2%；

3- external forms-hydroxyl -8- azepines bridge [3.2.1] octane -8- t-butyl formates：1H NMR(400MHz,CDCl₃)δ 1.47(9H,s),1.60-1.62(4H,m),1.94-1.99(4H,m),4.06-4.14(1H,m),4.19-4.28(2H,m).

3- inner mold-hydroxyl -8- azepines bridge [3.2.1] octane -8- t-butyl formates：1H NMR(400MHz,CDCl₃)δ 1.46(9H,s),1.69-1.73(2H,m),1.92-1.95(2H,m),2.02-2.17(4H,m),4.14-4.21(3H,m).

Step 3:External form -3- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -8- azepines It is prepared by bridge [3.2.1] octane -8 t-butyl formate

Under nitrogen protection, to equipped with 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (B-H) (500mg, 1.65mmol), 3- inner mold-hydroxyl -8- azepines bridge [3.2.1] octane -8- t-butyl formates (568mg, 2.50mmol) And PPh₃DIAD (667mg, 3.30mmol), the reaction is slowly added dropwise in (0 DEG C) in the 20mLTHF solution of (865mg, 3.30mmol) 3h is stirred at room temperature in mixture, and water quenching is then slowly added dropwise again and goes out reaction, and EA extracts (50mL x 3), and organic phase is with anhydrous Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=60：1~10:1) purifying obtains 600mg target chemical combination Thing, is white solid, yield 70.9%；

Inner mold -3- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -8- azepine bridges The preparation of [3.2.1] t-butyl formate of octane -8.

Method is same as above, and is white solid, yield 62.7%；

Step 4:External form -8- azepines bridge [3.2.1] octane -3- bases) -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] It is prepared by pyrimidine -4- amine

At room temperature, to equipped with external form -3- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- Base) -8- azepines bridge [3.2.1] octane -8 t-butyl formate (512mg, 1.00mmol) 20mL DCM solution in be slowly added dropwise TFA (2mL), the reactant mixture are stirred at room temperature reaction 2h, NaHCO are then slowly added dropwise again₃(1mol/L) is more than to pH 7, organic phase is separated, aqueous phase extracts (20mL x 3) with DCM, merges above-mentioned organic phase anhydrous Na₂SO₄Fully dry, vacuum is steamed Pass through chromatography (DCM after hair：MeOH=10：1~1:1) purifying obtains 250mg target compounds, is white solid, yield is 60.7%；

Inner mold -8- azepines bridge [3.2.1] octane -3- bases) -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- It is prepared by amine

Method is same as above, and is white solid, yield 30.6%；

Step 5:External form -3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1-yl) -8- azepines Bridge [3.2.1] octane -8- bases) propane -2- alkene -1- ketone preparation

Under nitrogen protection, to equipped with external form -8- azepines bridge [3.2.1] octane -3- bases) -3- (4- Phenoxyphenyls) - 1H- pyrazoles [3,4-d] pyrimidine -4- amine (103mg, 0.25mmol), TEA (51mg, 0.50mmol) 20mL DCM solution in (0 DEG C) acryloyl chloride (22mg, 0.24mmol) is slowly added dropwise, the reactant mixture is stirred at room temperature 3h, is then slowly added dropwise again Saturation NH₄Reaction is quenched in Cl, is washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄Fully dry, pass through after vacuum evaporation Chromatography (DCM：MeOH=50:1~20：1) purifying obtains 60mg target compounds, is white solid, yield 51.3%；

1H NMR(400MHz,d₆-DMSO)δ1.91-2.02(5H,m),2.07-2.27(3H,m),4.68-4.69(2H, M), 4.84-4.87 (1H, m), 5.26-5.35 (1H, m), 5.73 (1H, dd, J=10.4Hz, 2.4Hz), 6.23 (1H, dd, J= 16.4Hz, 2.4Hz), 6.79 (1H, dd, J=16.4Hz, 10.4Hz), 7.11-7.20 (6H, m), 7.41-7.45 (2H, m), 7.62 (2H, d, J=8.4Hz), 8.26 (1H, s)

EM (calculated value)：466.2；MS(ESI)m/e(M+1H)⁺：467.2

Inner mold -3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1-yl) -8- azepine bridges [3.2.1] octane -8- bases) propane -2- alkene -1- ketone (2) preparation

Method is same as above, and is white solid, yield 48.5%；

1H NMR(400MHz,d₆-DMSO)δ1.81-1.87(1H,m),1.92-2.09(3H,m),2.33-2.46(4H, M), 4.60-4.63 (2H, m), 4.84-4.87 (1H, m), 5.73 (1H, dd, J=10.4Hz, 2.4Hz), 6.23 (1H, dd, J= 16.4Hz, 2.4Hz), 6.78 (1H, dd, J=16.4Hz, 10.4Hz), 6.85-6.92 (2H, m), 7.12-7.21 (6H, m), 7.42-7.46 (2H, m), 7.68 (2H, d, J=8.4Hz), 8.23 (1H, s)

EM (calculated value)：466.2；MS(ESI)m/e(M+1H)⁺：467.2

Embodiment 3

1- ((1R, 5S) -3- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -9- azepines Bridge [3.3.1] nonane -9- bases) propyl- 2- alkene -1- ketone preparation

Synthesis step is as follows：

Step 1：The preparation of 9- benzyl -9- azabicyclos [3.3.1] nonyl- 3- ketone

Under ice bath, to equipped with benzylamine hydrochloride (3.8g, 26.46mmol), glutaraldehyde (25% glutaraldehyde water solution, 8.8mL, 21.90mmol) 20mL water in add 1,3- acetone dicarboxylic acids (3.2g, 21.90mmol) and 10% sodium acetate (7.5mL), removes ice bath afterwards, and 2h is stirred at room temperature in the reactant mixture, 12h is then stirred at 50 DEG C, then slowly Reinforcing body NaHCO₃It is more than 7 to pH, separates organic phase, aqueous phase extracts (20mL x 3) with DCM, merges above-mentioned organic phase with anhydrous Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=10：1) purifying obtains 2.3g target compounds, for white Solid, yield 71.9%；

Step 2：The preparation of 3- carbonyl -9- azabicyclos [3.3.1] nonane -9- t-butyl formates

At room temperature, to the 80mL equipped with 9- benzyl -9- azabicyclos [3.3.1] nonyl- 3- ketone (2.3g, 10.03mmol) (Boc) is added in EA solution₂O (2.3g, 10.53mmol), 10% palladium carbon (250mg), the reactant mixture is under an atmosphere of hydrogen (2atm) is stirred overnight at room temperature, is filtered after the completion of reaction and is removed palladium carbon, passes through chromatography (PE after vacuum evaporation：EA=10：1) Purifying obtains 1.7g target compounds, is white solid, yield 70.8%；

Step 3：The preparation of 3- hydroxyl -9- azabicyclos [3.3.1] nonane -9- t-butyl formates

At room temperature, to equipped with 3- carbonyl -9- azabicyclos [3.3.1] nonane -9- t-butyl formates (1.7g, NaBH is added portionwise in 25mL THF solutions 7.10mmol)₄(537mg, 14.20mmol), the reactant mixture is at room temperature 1h is stirred, saturation NH is then slowly added dropwise₄Reaction is quenched in Cl, and (40mL x 3), anhydrous Na are extracted with EA₂SO₄Fully dry, very Pass through chromatography (PE after sky evaporation：EA=10：1~3:1) purifying obtains 1.45g target compounds, is colorless oil, yield 84.8%；

1H NMR(400MHz,CDCl₃)δ1.29-1.38(2H,m),1.45(9H,s),1.50-1.68(5H,m),2.03- 2.14(1H,m),2.27-2.38(2H,m),3.66-3.70(1H,m),4.35-4.38(1H,m),4.48-4.50(1H,m).

Step 4：3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -9- azabicyclos The preparation of [3.3.1] nonane -9- t-butyl formates

Under nitrogen protection, to equipped with 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (B-H) (1.2g, 3.96mmol), 3- hydroxyl -9- azabicyclos [3.3.1] nonane -9- t-butyl formates (1.5g, 6.22mmol) and DIAD (1.6g, 7.92mmol) is slowly added dropwise in (0 DEG C) in PPh3 (2.1g, 7.92mmol) 20mL THF solutions, and the reaction mixes 3h is stirred at room temperature in compound, and water quenching is then slowly added dropwise and goes out reaction, EA extracts (50mL x 3), organic phase anhydrous Na₂SO₄ Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=60：1~10:1) purifying obtains 800mg target compounds, is White solid, yield 38.1%；

Step 5：1- (9- azabicyclos [3.3.1] nonane -3- bases) -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] The preparation of pyrimidine -4- amine

At room temperature, to equipped with 3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -9- TFA is slowly added dropwise in the 20mL DCM solution of azabicyclo [3.3.1] nonane -9- t-butyl formates (700mg, 1.33mmol) (2mL), the reactant mixture are stirred at room temperature 2h, NaHCO are then slowly added dropwise₃(1mol/L) is more than 7 to pH, separates organic Phase, aqueous phase extract (20mL x 3) with DCM, merge above-mentioned organic phase anhydrous Na₂SO₄Fully dry, pass through color after vacuum evaporation Spectrometry (DCM：MeOH=10：1~1:1) purifying obtains 450mg target compounds, is white solid, yield 79.4%；

Step 6：1- (3- (4- amine -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -9- azabicyclos [3.3.1] nonane -9- bases) propane -2- alkene -1- ketone preparation

Under nitrogen protection, to equipped with 1- (9- azabicyclos [3.3.1] nonane -3- bases) -3- (4- Phenoxyphenyls) - 1H- pyrazoles [3,4-d] pyrimidine -4- amine (400mg, 0.94mmol), TEA (190mg, 1.88mmol) 20mL DCM solution in (0 DEG C) acryloyl chloride (81mg, 0.89mmol) is slowly added dropwise, the reactant mixture is stirred at room temperature 3h, states reaction then up Saturation NH is slowly added dropwise in mixture₄Reaction is quenched in Cl, is washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄It is fully dry It is dry, pass through chromatography (DCM after vacuum evaporation：MeOH=50:1~20：1) purifying obtains 90mg target compounds, solid for white Body, yield 19.9%.

1H NMR(400MHz,d₆-DMSO)δ1.77-1.92(5H,m),2.03-2.13(3H,m),2.30-2.38(2H, M), 4.51 (1H, s), 4.86 (1H, s), 5.70 (1H, dd, J=10.4Hz, 2.4Hz), 5.77-5.85 (1H, m), 6.17 (1H, Dd, J=16.4Hz, 2.4Hz), 6.87 (1H, dd, J=16.4Hz, 10.4Hz), 7.10-7.21 (6H, m), 7.41-7.45 (2H, m), 7.64 (2H, d, J=8.8Hz), 8.27 (1H, s)

EM (calculated value)：480.2；MS(ESI)m/e(M+1H)⁺：481.3

Embodiment 4

1- (3- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxa- -8- azepines Spiral shell [4.5] certain herbaceous plants with big flowers alkane -8- bases) propyl- 2- alkene -1- ketone preparation

Synthesis step is as follows：

Step 1:The preparation of the hydroxy piperidine -1- t-butyl formates of 4- pi-allyls -4

At room temperature in nitrogen atmosphere, slowly dripped into the 20mL dry diethyl ether solution equipped with magnesium powder (3.2g, 13.33mmol) Add a small amount of allyl bromide, bromoallylene, the reactant mixture heating stirring, wait reaction after triggering to continue that allyl bromide, bromoallylene is slowly added dropwise, rate of addition Control is cooled to -15 DEG C in maintenance reaction reflux state, altogether addition allyl bromide, bromoallylene (13.3g, 0.11mol), reaction after completing, The THF solution (100mL) of N- tertbutyloxycarbonyl -4- piperidones (10.0g, 50.19mmol) is slowly added into, reaction system is in room temperature Lower stirring 5h, reaction are cooled to 0 DEG C after completing, are then slowly added into saturation NH₄Cl extracts reaction of going out, and separates organic phase, aqueous phase (30mL x 3) is extracted with EA, merges above-mentioned organic phase anhydrous Na₂SO₄Fully dry, 12g targeteds are obtained after vacuum evaporation Compound, it is pale yellow oil, yield 98.9%, gained target compound is directly used in react in next step, without further Purifying；

Step 2:The preparation of 3- hydroxyls -1- oxa- -8- azaspiros [4.5] decane -8- t-butyl formates

At room temperature, to equipped with the hydroxy piperidine -1- t-butyl formates (11.3g, 46.82mmol) of 4- pi-allyls -4 NaIO is slowly added into the mixed solution of the 100mL tert-butyl alcohols and 40mL water₄(11.1g, 51.51mmol), the reactant mixture exist 0.5h is stirred at 50 DEG C, Na is then slowly added dropwise₂S₂O₅The aqueous solution 30mL of (9.8g, 51.55mmol), reactant mixture is 50 7h is reacted at DEG C again, 48h is stirred at room temperature, water is then slowly added into again and extracts reaction of going out, EA extractions (50mL x 3), merging is above-mentioned to be had Machine mutually uses anhydrous Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=10：1~1:1) purifying obtains 2.5g mesh Compound is marked, is yellow oil, yield 20.8%；

1H NMR(400MHz,CDCl₃)δ1.46(9H,s),1.48-1.53(2H,m),1.4-1.56(2H,m),1.64- 1.71 (2H, m), 1.96 (1H, dd, J=13.6Hz, 6.4Hz), 3.3-3.38 (2H, m), 3.59-3.60 (1H, m), 3.80- 3.83 (1H, m), 3.92 (1H, dd, J=10.0Hz, 4.0Hz), 4.50-4.52 (1H, m)

Step 3:3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxa- -8- nitrogen The preparation of miscellaneous loop coil [4.5] decane -8- t-butyl formates

Under nitrogen protection, to equipped with 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (B-H) (1.2g, 3.96mmol), 3- hydroxyls -1- oxa- -8- azaspiros [4.5] decane -8- t-butyl formates (1.6g, 6.22mmol) And PPh₃DIAD (1.6g, 7.92mmol), the reaction is slowly added dropwise in (0 DEG C) in the 20mL THF solutions of (2.1g, 7.92mmol) 3h is stirred at room temperature in mixture, and water quenching is then slowly added dropwise again and goes out reaction, and EA extracts (50mL x 3), and organic phase is with anhydrous Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=60：1~10:1) purifying obtains 1.3g target chemical combination Thing, is white solid, yield 60.5%；

1H NMR(400MHz,d₆-DMSO)δ1.47(9H,s),1.58-1.83(4H,m),2.36-2.38(2H,m), 3.28-3.29(2H,m),3.42-3.51(2H,m),4.04-4.08(1H,m),4.20-4.24(1H,m),5.53-5.57(1H, M), 7.12-7.21 (6H, m), 7.42-7.46 (3H, m), 7.66 (3H, d, J=8.4Hz), 8.25 (1H, s)

Step 4:3- (4- Phenoxyphenyls) -1- (1- oxa- -8- azaspiros [4.5] decane -3- bases) -1H- pyrazoles [3,4- D] pyrimidine -4- amine preparation

At room temperature, to equipped with 3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- Slowly it is added dropwise in the 40mL DCM solution of oxa- -8- azaspiros [4.5] decane -8- t-butyl formates (1.5g, 2.76mmol) TFA (5mL), the reactant mixture are stirred at room temperature 2h, NaHCO are then slowly added dropwise₃(1mol/L) is more than 7 to pH, separates Organic phase, aqueous phase extract (20mL x 3) with DCM, merge above-mentioned organic phase anhydrous Na₂SO₄Fully dry, lead to after vacuum evaporation Cross chromatography (DCM：MeOH=10：1~1:1) purifying obtains 1.0g target compounds, is white solid, yield 82.0%； EM (calculated value)：442.2；MS(ESI)m/e(M+1H)⁺：443.2

Step 5:1- (3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1-yl) -1- oxa-s - 8- azaspiros [4.5] decane -8-yl) propane -2- alkene -1- ketone preparation

Under nitrogen protection, to equipped with 3- (4- Phenoxyphenyls) -1- (1- oxa- -8- azaspiros [4.5] decane -3- Base) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (751mg, 1.70mmol), TEA (344mg, 3.40mmol) 20mL DCM solution In (0 DEG C) acryloyl chloride (139mg, 1.53mmol) is slowly added dropwise, 3h is stirred at room temperature in the reactant mixture, then again slowly Saturation NH is added dropwise₄Reaction is quenched in Cl, is washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄Fully dry, after vacuum evaporation Pass through chromatography (DCM：MeOH=30：1) purifying obtains 350mg target compounds, is white solid, yield 41.5%；

1H NMR(400MHz,d₆- DMSO) δ 1.66-1.94 (4H, m), 2.44 (2H, d, J=7.2Hz), 3.29-3.26 (1H, m), 3.57-3.59 (1H, m), 3.73-3.77 (1H, m), 3.89-3.91 (1H, m), 4.14 (1H, dd, J=8.8Hz, 6.0Hz), 4.28-4.32 (1H, m), 5.60-5.64 (1H, m), 5.71 (1H, d, J=10.0Hz), 6.14 (1H, dd, J= 16.8Hz, 2.0Hz), 6.85-6.91 (1H, m), 7.18-7.27 (5H, m), 7.49 (2H, t, J=8.0Hz), 7.72 (2H, d, J =8.8Hz), 8.31 (1H, s)

EM (calculated value)：496.2；MS(ESI)m/e(M+1H)⁺：497.3

Embodiment 5

1- (3- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1-yl) -4- hydroxyls -3,4- two Hydrogen quinoline -1 (2H)-yl) propyl- 2- alkene -1- ketone preparation

Synthesis step is as follows：

Step 1：The preparation of quinoline -1 (2H)-carboxylic acid tert-butyl ester

Under nitrogen protection, slowly it is added dropwise at 0 DEG C in equipped with quinoline (9.0g, 69.68mmol) 70mL THF solutions DIBAl (76.36mL, 76.6mmol, 1mol/L THF solution), reaction temperature is kept to be less than 45 DEG C during dropwise addition, room temperature is anti-afterwards Answer 1h, TLC detections to add TEA (12mL, 86.50mmol) into above-mentioned reactant mixture after completing, then be added dropwise to (Boc)₂O The toluene solution 40mL of (19.0g, 87.06mmol), also keep during addition reaction temperature to be less than 45 DEG C, continue afterwards to reactant DMAP (8.5g, 69.58mmol) is added in system and stirs 2h at room temperature, TCL detection reactions are completed to adjust pH value with dilute HCl later (50mL x 3) is extracted to 3~5, EA, passes through chromatography (PE after vacuum evaporation：EA=200：1) purifying obtains 2.0g targeteds Compound, is pale yellow oil, yield 12.4%；

1H NMR(400MHz,CDCl₃)δ1.52(9H,s),4.35-4.37(2H,m),5.97-6.01(1H,m),6.47 (1H, d, J=8.4Hz), 7.03-7.05 (2H, m), 7.15-7.19 (1H, m), 7.55 (1H, d, J=8.0Hz)

Step 2：The preparation of bromo- 3- hydroxyls -3,4- EEDQs -1 (the 2H)-carboxylic acid tert-butyl esters of 4-

At room temperature, to the 53mL DMSO equipped with quinoline -1 (2H)-carboxylic acid tert-butyl ester (3.3g, 14.27mmol) and NBS (3.8g, 21.35mmol) is added portionwise in the mixed solution of 0.53mL water, 0.5h is stirred at room temperature in the reactant mixture, Then NaHCO is slowly added dropwise₃(1mol/L) is more than 7, EA extractions (40mL x 3) to pH, merges above-mentioned organic phase with anhydrous Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=30：1~10:1) purifying obtains 1.0g target compounds, For yellow oil, yield 46.6%；

1H NMR(400MHz,CDCl₃) δ 1.40 (9H, s), 2.95 (1H, d, J=14.4Hz), 3.78-3.83 (2H, m), 4.75 (1H, d, J=14.4Hz), 7.02-7.06 (2H, m), 7.19-7.24 (1H, m), 7.29-7.31 (2H, m)

Step 3：3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -4- hydroxyls -3,4- The preparation of EEDQ -1 (2H)-carboxylic acid tert-butyl ester

Reactant mixture 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (B-H) (1.2g, 3.96mmol), bromo- 3- hydroxyls -3, the 4- EEDQs -1 (2H) of 4--carboxylic acid tert-butyl ester (1.3g, 3.96mmol), K₂CO₃(1.6g, 11.88mmol) it is stirred overnight in 15mL DMF in 90 DEG C, reaction is cooled to room temperature after completing, to above-mentioned reactant mixture Middle addition water (50mL), EA extractions (40mL x 3), merge above-mentioned organic phase and use with water and saturated common salt washing, organic phase nothing Water Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=70：1~60:1) purifying obtains 504mg targeteds Compound, is white solid, yield 23.1%；

1H NMR(400MHz,d₆- DMSO) δ 1.50 (9H, s), 3.64 (1H, dd, J=13.2Hz, 9.2Hz), 4.25 (1H, dd, J=12.8Hz, 4.0Hz), 4.41-4.45 (1H, m), 5.69 (1H, d, J=5.2Hz), 5.87 (1H, d, J= 7.2Hz), 6.64 (1H, d, J=7.6Hz), 6.94 (1H, t, J=7.6Hz), 7.09-7.22 (6H, m), 7.40-7.44 (2H, m),7.60-7.67(3H,m),8.31(1H,s).

Step 4：3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1,2,3,4- four The preparation of hydrogen quinoline -4- alcohol

At room temperature, to equipped with 3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -4- 5mL is added in the 100mL round-bottomed flasks of hydroxyl -3,4- EEDQ -1 (2H)-carboxylic acid tert-butyl ester (445mg, 0.81mmol) DCM solution is completely dissolved it, then adds 60mL saturations HCl EA solution, and the reactant mixture was stirred at room temperature At night, it is evaporated in vacuo after the completion of reaction to dry, then is dissolved with 100mL DCM, this DCM organic phase uses saturation NaHCO respectively₃And saturation Brine It, organic phase anhydrous Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=70：1~65: 1) purifying obtains 216mg target compounds, is faint yellow solid, yield 59.2%；

1H NMR(400MHz,d₆- DMSO) δ 3.22 (1H, t, J=10.8Hz), 3.42-3.47 (1H, m), 4.46-4.51 (1H, m), 5.31 (1H, d, J=5.2Hz), 5.84 (1H, d, J=8.4Hz), 5.97 (1H, d, J=2.4Hz), 6.23 (1H, d, ), J=7.6Hz 6.35 (1H, t, J=7.6Hz), 6.55 (1H, J=7.6Hz), 6.90 (1H, t, J=7.6Hz), 7.09-7.19 (5H,m),7.40-7.44(2H,m),7.61-7.63(2H,m),8.28(1H,s).

Step 5：1- (3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -4- hydroxyls - 3,4- EEDQs -1 (2H)-yl) propane -2- alkene -1- ketone preparation

Under nitrogen protection, to equipped with 3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- Base) -1,2,3,4- tetrahydroquinoline -4- alcohol (300mg, 0.67mmol), Cs₂CO₃The 20mL DCM solution of (437mg, 1.34mmol) In (0 DEG C) acryloyl chloride (61mg, 0.67mmol) is slowly added dropwise, the reactant mixture is stirred at room temperature 2h, then slowly drips Add saturation NH₄Reaction is quenched in Cl, is washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄Fully dry, lead to after vacuum evaporation Cross chromatography (DCM：MeOH=25：1) purifying obtains 30mg target compounds, is faint yellow solid, yield 8.9%.

1H NMR(400MHz,d₆- DMSO) δ 3.84 (1H, dd, J=12.8Hz, 7.6Hz), 4.29-4.33 (1H, m), 4.50-4.52 (1H, m), 5.77-5.82 (2H, m), 5.88 (1H, d, J=6.8Hz), 6.29 (1H, dd, J=16.8Hz, 2.0Hz), 6.67 (1H, dd, J=16.0Hz, 10.0Hz), 6.76 (1H, d, J=7.6Hz), 7.06-7.19 (7H, m), 7.26- 7.29 (2H, m), 7.40-7.44 (2H, m), 7.62 (2H, d, J=8.4Hz), 8.29 (1H, s)

EM (calculated value)：504.2；MS(ESI)m/e(M+1H)⁺：505.2

Embodiment 6

1- (4- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) spiral shell [chroman -2,4'- piperazines Pyridine] -1'- bases) propyl- 2- alkene -1- ketone synthesis step it is as follows：

Step 1：The preparation of 4- carbonyls spiral shell [chroman -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters

At room temperature, N- is sequentially added into the 75mL methanol solutions equipped with o-hydroxyacetophenone (6.8g, 49.94mmol) Tertbutyloxycarbonyl -4- piperidones (10.0g, 49.94mmol), nafoxidine (3.6g, 49.94mmol), afterwards this reaction mixing Thing is stirred at room temperature overnight, and reaction is gone out after completing with HCl (1mol/L) extractions, and extracting (100mL x3) merging organic phase with EA uses Anhydrous Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=20：1~10:1) purifying obtains 11.1g targeteds Compound, is colorless oil, yield 70.0%；

Step 2：The preparation of 4- hydroxyls spiral shell [chroman -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters

At room temperature, to equipped with 4- carbonyls spiral shell [chroman -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters (4.8g, 15.13mmol) 37.5mL THF and 15mL MeOH mixed solution in NaBH is added portionwise₄(1.1g, 30.26mmol), the reactant mixture 1h is stirred at room temperature, saturation NH is then slowly added dropwise₄Reaction is quenched in Cl, and (40mL x 3), anhydrous Na are extracted with EA₂SO₄Fully Dry, pass through chromatography (PE after vacuum evaporation：EA=20：1~3:1) purifying obtains 3.8g target compounds, is yellow oily Thing, yield 79.2%；

1H NMR(400MHz,CDCl₃)δ1.46(9H,s),1.49-1.70(3H,m),1.78-1.97(4H,m),2.14 (1H, d, J=13.6Hz), 3.17-3.26 (2H, m), 3.83-3.86 (2H, m), 4.88 (1H, q, J=6.8Hz), 6.85 (1H, D, J=8.0Hz), 6.96 (1H, t, J=7.2Hz), 7.20 (1H, t, J=7.2Hz), 7.43 (1H, d, J=7.6Hz)

EM (calculated value)：319.2；MS(ESI)m/e(M+1H)⁺：320.3

Step 3：4- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) spiral shell [chroman -2, 4'- piperidines] -1'- carboxylic acid tert-butyl esters preparation

Under nitrogen protection, to equipped with 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (B-H) (1.5g, 4.95mmol), 4- hydroxyls spiral shell [chroman -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters (3.2g, 9.90mmol) and PPh₃ DIAD (1.5g, 7.43mmol), the reactant mixture is slowly added dropwise in (0 DEG C) in the 90mL THF solutions of (2.3g, 7.43mmol) 3h is stirred at room temperature, water quenching is then slowly added dropwise again and goes out reaction, DCM extracts (50mL x 3), organic phase anhydrous Na₂SO₄Fill Divide drying, pass through chromatography (DCM after vacuum evaporation：MeOH=100:1) purifying obtains 880mg target compounds, solid for white Body, yield 29.1%；EM (calculated value)：604.3；MS(ESI)m/e(M+1H)⁺：605.3；

Step 4：3- (4- Phenoxyphenyls) -1- (spiral shell [chroman -2,4'- piperidines] -4- bases) -1H- pyrazoles [3,4-d] is phonetic The preparation of pyridine -4- amine

At room temperature, to equipped with 4- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) spiral shell It is molten that 10mL DCM are added in the 150mL round-bottomed flasks of [chroman -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters (850mg, 1.41mmol) Liquid is completely dissolved it, then adds 100mL saturations HCl EA solution, and the reactant mixture is stirred at room temperature overnight, instead Should after the completion of be evaporated in vacuo to dry, then dissolved with 100mL DCM, this DCM organic phase uses saturation NaHCO respectively₃And saturated common salt Water washing, organic phase anhydrous Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=40：1~5:1) it is pure Change obtains 570mg target compounds, is faint yellow solid, yield 80.2%；

Step 5：1- (4- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) spiral shell [chroman - 2,4'- piperidines] -1'- or so) propyl- 2- alkene -1- ketone preparation

Under nitrogen protection, to equipped with 3- (4- Phenoxyphenyls) -1- (spiral shell [chroman -2,4'- piperidines] -4- bases) -1H- pyrroles Azoles [3,4-d] pyrimidine -4- amine (564mg, 1.12mmol), TEA (226mg, 2.24mmol) 20mL DCM solution in (0 DEG C) it is slow Slow that acryloyl chloride (101mg, 1.12mmol) is added dropwise, the reactant mixture is stirred at room temperature 3h, saturation is then slowly added dropwise again NH₄Reaction is quenched in Cl, is washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄Fully dry, pass through chromatogram after vacuum evaporation Method (DCM：MeOH=60：1) purifying obtains 350mg target compounds, is white solid, yield 55.9%.

1H NMR(400MHz,d₆- DMSO) δ 1.72-1.83 (2H, m), 1.89-1.99 (2H, m), 2.38 (1H, q, J= 6.4Hz),2.68-2.74(1.5H,m),3.00-3.07(0.5H,m),3.58-3.64(1H,m),3.88-3.98(1H,m), 4.18-4.27 (1H, m), 5.69 (1H, dd, J=10.4Hz, 2.4Hz), 6.12 (1H, dd, J=16.4Hz, 2.0Hz), 6.17- 6.20 (1H, m), 6.47 (1H, d, J=7.6Hz), 6.74-6.78 (1H, m), 6.79-6.88 (1H, m), 6.92 (1H, d, J= 8.0Hz),7.09-7.19(7H,m),7.39-7.44(2H,m),7.61-7.63(2H,m),8.31(1H,s).

EM (calculated value)：558.2；MS(ESI)m/e(M+1H)⁺：559.3

Embodiment 7

1- (4- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -7- fluorine spiral shell [chroman -2, 4'- piperidines] -1'- bases) propyl- 2- alkene -1- ketone synthesis step it is as follows：

Step 1:The preparation of the fluoro- 4- carbonyls spiral shells of 7- [chroman -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters

At room temperature, into the 75mL methanol solutions equipped with 4- fluorin-2-hydroxyacetophenones (3.4g, 20.76mmol) successively N- tertbutyloxycarbonyl -4- piperidones (4.4g, 20.76mmol), nafoxidine (1.6g, 20.76mmol) are added, this reacts afterwards Mixture is stirred at room temperature overnight, and reaction is gone out after completing with HCl (1M) extractions, and extracting (100mL x3) merging organic phase with EA uses Anhydrous Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=20：1~10:1) purifying obtains 6.5g target chemical combination Thing, is colorless oil, yield 88.2%；

Step 2:The preparation of the fluoro- 4- hydroxyls spiral shells of 7- [chroman -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters

At room temperature, to equipped with the fluoro- 4- carbonyls spiral shells of 7- [chroman -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters (2.8g, NaBH is added portionwise in the mixed solution of 25mL THF and 10mL MeOH 8.35mmol)₄(629mg, 16.64mmol), this is anti- Answer mixture that 1h is stirred at room temperature, saturation NH is then slowly added dropwise₄Reaction is quenched in Cl, is extracted (30mL x 3) with EA, anhydrous Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=20：1~3:1) purifying obtains 2.8g target compounds, is Rice-pudding color grease, yield 97.2%；

Step 3:4- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -7- fluorine spiral shell [colors Full -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters preparation

Under nitrogen protection, to equipped with 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (B-H) The fluoro- 4- carbonyls spiral shell of (2.0g, 6.59mmol), 7- [chroman -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters (5.6g, 16.48mmol) and PPh₃DIAD (2.0g, 10.43mmol), the reaction is slowly added dropwise in (0 DEG C) in the 90mL THF solutions of (2.7g, 10.43mmol) 3h is stirred at room temperature in mixture, and water quenching is then slowly added dropwise again and goes out reaction, and DCM extracts (50mL x 3), and organic phase is with anhydrous Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=100:1~50:1) purifying obtains 1.8g target chemical combination Thing, is faint yellow solid, yield 43.9%；EM (calculated value)：622.3；MS(ESI)m/e(M+1H)⁺：623.3；

Step 4:1- (7- fluorine spiral shell [chroman -2,4'- piperidines] -4- bases) -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] The preparation of pyrimidine -4- amine

At room temperature, to equipped with 4- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -7- 10mL is added in the 150mL round-bottomed flasks of fluorine spiral shell [chroman -2,4'- piperidines] -1'- carboxylic acid tert-butyl esters (1.5g, 2.41mmol) DCM solution is completely dissolved it, then adds 100mL saturations HCl EA solution, and the reactant mixture was stirred at room temperature At night, it is evaporated in vacuo after the completion of reaction to dry, then is dissolved with 100mL DCM, this DCM organic phase uses saturation NaHCO respectively₃And saturation Brine It, organic phase anhydrous Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=10：1~5: 1) purifying obtains 824mg target compounds, is faint yellow solid, yield 65.4%；

Step 5:1- (4- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1-yl) -7- fluorine spiral shells [chroman -2,4'- piperidines] -1'- bases) propane -2- alkene -1- ketone preparation

Under nitrogen protection, to equipped with 1- (7- fluorine spiral shell [chroman -2,4'- piperidines] -4- bases) -3- (4- Phenoxyphenyls) - 1H- pyrazoles [3,4-d] pyrimidine -4- amine (597mg, 1.14mmol), TEA (231mg, 2.28mmol) 20mL DCM solution in (0 DEG C) acryloyl chloride (103mg, 1.14mmol) is slowly added dropwise, the reactant mixture is stirred at room temperature 3h, states reaction then up Saturation NH is slowly added dropwise in mixture₄Reaction is quenched in Cl, is washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄It is fully dry It is dry, pass through chromatography (DCM after vacuum evaporation：MeOH=60：1~40：1) purifying obtains 380mg target compounds, solid for white Body, yield 57.8%.

1H NMR(400MHz,d₆- DMSO) δ 1.71-1.83 (2H, m), 1.90-1.99 (2H, m), 2.40 (1H, q, J= 6.4Hz),2.67-2.72(1H,m),3.01-3.07(0.5H,m),3.24-3.27(0.5H,m),3.57-3.62(1H,m), 3.88-3.97 (1H, m), 4.18-4.26 (1H, m), 5.69 (1H, dd, J=10.4Hz, 2.4Hz), 6.10-6.18 (2H, m), 6.49-6.52 (1H, m), 6.62 (1H, ddd, J=11.2Hz, 8.8Hz, 2.4Hz), 6.79-6.91 (2H, m), 7.09-7.20 (6H,m),7.40-7.44(2H,m),7.61-7.63(2H,m),8.31(1H,s).

EM (calculated value)：622.3；MS(ESI)m/e(M+1H)⁺：623.3.

Embodiment 8

1- (3- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxa- -7- azepines Spiral shell [4.4] nonyl- 7- yls) propyl- 2- alkene -1- ketone synthesis step it is as follows：

Step 1：The preparation of 3- pi-allyl -3- hydroxypyrrole -1- carboxylic acid tert-butyl esters

Under nitrogen protection, to equipped with 1- t-butoxycarbonyl -3- pyrrolidones (5.0g, 26.99mmol), allyl bromide, bromoallylene (0 DEG C) adds saturation NH in the 20mL THF solutions of (8.2g, 67.48mmol)₄Cl (28mL), then add in batches at this temperature Enter zinc powder (3.8g, 67.48mmol), keep reaction temperature to be no more than 40 DEG C in adition process, the reactant mixture is in room temperature Lower stirring 12h, TLC detection reaction are completed, and 2mol/L H are then slowly added dropwise again₂SO₄Reaction is quenched, EA extractions (40mLx3), closes And washed after organic phase with saturation NaCl, anhydrous Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=1：1) it is pure Change obtains 3.8g target compounds, is white solid, yield 62.0%；

1H NMR(400MHz,CDCl₃)δ1.46(9H,s),1.84-1.87(2H,m),1.92-1.95(1H,m),2.40 (2H, d, J=7.6Hz), 3.24-3.39 (2H, m), 3.46-3.53 (2H, m), 5.18-5.22 (2H, m), 5.84-5.90 (1H, m).

Step 2：The preparation of 3- (2,3- dibromopropyls) -3- hydroxypyrrole -1- carboxylic acid tert-butyl esters

Under nitrogen protection, to equipped with 3- pi-allyl -3- hydroxypyrrole -1- carboxylic acid tert-butyl esters (1.0g, 4.40mmol) Br is slowly added dropwise at -15 DEG C in 10mL DCM solution₂(703mg, 4.40mmol), 0.5h, TLC detections is stirred at room temperature again afterwards After reaction is completed, saturation Na is slowly added into₂S₂O₃Extract reaction of going out, DCM extractions (30mL x3), saturation is used after merging organic phase NaHCO₃Washing, anhydrous Na₂SO₄Fully dry, obtain 1.0g target compound crude products to dry after vacuum evaporation, be yellow oily Thing, being directly used in reaction in next step need not be further purified；

Step 3：The preparation of bromo- 1- carbonyls -7- azaspiros [4.4] nonane -7- carboxylic acid tert-butyl esters of 3-

Under nitrogen protection, to the 10mL equipped with 3- (2,3- dibromopropyl) -3- hydroxypyrrole -1- carboxylic acid tert-butyl esters Solid K is added in MeOH solution at room temperature₂CO₃(857mg, 6.21mmol), is stirred overnight at room temperature again afterwards, TLC detection reactions After completion, water is added into above-mentioned reactant mixture and extracts reaction of going out, DCM extractions (20mL x 3), nothing is used after merging organic phase Water Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=10：1) purifying obtains 340mg target compounds, is light Yellow solid, yield 25.2%；

Step 4：3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxo -7- nitrogen The preparation of miscellaneous spiral shell [4.4] nonane -7- carboxylic acid tert-butyl esters

Reactant mixture 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (B-H) (600mg, 1.98mmol), bromo- 1- carbonyls -7- azaspiros [4.4] nonane -7- carboxylic acid tert-butyl esters (600mg, 1.98mmol) of 3-, Cs₂CO₃ (2.0g, 5.94mmol) is stirred overnight in 20mL DMF in 85 DEG C, and reaction is cooled to room temperature after completing, and is mixed to above-mentioned reaction Water (40mL) is added in compound, EA extractions (30mL x 3), merges above-mentioned organic phase and uses with water and saturated common salt washing, organic phase Use anhydrous Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=40：1~15:1) purifying obtains 304mg mesh Compound is marked, is white solid, yield 29.3%；

Step 5：3- (4- Phenoxyphenyls) -1- (1- oxo -7- azaspiros [4.4] nonane -3- bases) -1H- pyrazoles [3,4- D] pyrimidine -4- amine preparation

At room temperature, to equipped with 3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- 60mL is added in the 500mL round-bottomed flasks of oxo -7- azaspiros [4.4] nonane -7- carboxylic acid tert-butyl esters (1.0g, 1.89mmol) DCM solution is completely dissolved it, then adds 200mL saturations HCl EA solution, and the reactant mixture was stirred at room temperature At night, it is evaporated in vacuo after the completion of reaction to dry, then is dissolved with 300mL DCM, this DCM organic phase uses saturation NaHCO respectively₃And saturation Brine It, organic phase anhydrous Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=10：1~5: 1) purifying obtains 650mg target compounds, is faint yellow solid, yield 80.1%；

Step 6：1- (3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxos - 7- azaspiros [4.4] nonane -7- bases) propane -2- alkene -1- ketone preparation

Under nitrogen protection, to equipped with 3- (4- Phenoxyphenyls) -1- (1- oxo -7- azaspiros [4.4] nonane -3- Base) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (300mg, 0.70mmol), TEA (141mg, 1.40mmol) 20mL DCM solution In (0 DEG C) acryloyl chloride (64mg, 0.70mmol) is slowly added dropwise, the reactant mixture is stirred at room temperature 3h, states then up Saturation NH4Cl is slowly added dropwise in reactant mixture reaction is quenched, washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄Fill Divide drying, pass through chromatography (DCM after vacuum evaporation：MeOH=60：1~40：1) purifying obtains 102mg target compounds, is white Color solid, yield 30.2%.

1H NMR(400MHz,d₆-DMSO)δ1.95-2.19(2H,m),2.53-2.67(2H,m),3.38-3.41(1H, m),3.44-3.51(1H,m),3.73-3.78(1H,m),3.80-3.83(0.5H,m),3.93-3.95(0.5H,m),4.03- 4.09 (1H, m), 4.22-4.28 (1H, m), 5.54-5.59 (1H, m), 5.64 (1H, ddd, J=12.4Hz, 10.0Hz, 2.0Hz), 6.12 (1H, ddd, J=11.6Hz, 9.2Hz, 2.0Hz), 6.49 (0.5H, dd, J=16.8Hz, 10.0Hz), 6.59 (0.5H, dd, J=16.8Hz, 10.4Hz), 7.12-7.21 (6H, m), 7.44 (2H, t, J=8.0Hz), 7.69 (2H, d, J= 8.4Hz),8.26(1H,s).

EM (calculated value)：682.2；MS(ESI)m/e(M+1H)⁺：483.3

Embodiment 9

1- (4- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1-yl) -2- azepine bridges [4.1.0] heptane -2- bases) propyl- 2- alkene -1- ketone synthesis step it is as follows：

Step 1：The preparation of (the 2H)-benzyl carboxylate of 3- hydroxyl -3,4- dihydropyridines -1

At -78 DEG C, to equipped with 3- pyridones (12.0g, 0.12mol), NaHCO₃(7.8g, 92.85mmol's) NaBH is slowly added portionwise in the anhydrous MeOH solution of 20mL₄(12.6g, 0.33mol), controlling reaction temperature are less than -60 DEG C, afterwards CbzCl (36.4mL, 1.9mol) is added dropwise to the reactant mixture at -78 DEG C, and stirs 1.5h at this temperature, TLC inspections Survey after reaction completion, 1mol/L NaOH are slowly added dropwise into above-mentioned reactant mixture reaction is quenched, EA extractions (50mLx4) are closed And washed after organic phase with saturation NaCl, anhydrous Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=5：1) it is pure Change obtains 11.9g target compounds, is white solid, yield 41.3%；

1H NMR(400MHz,CDCl₃)δ1.99-2.12(2H,m),2.35-2.39(1H,m),3.60-3.71(2H,m), 4.13-4.16 (1H, m), 4.78-4.88 (1H, m), 5.19 (2H, d, J=3.6Hz), 6.82-6.95 (1H, m), 7.32-7.38 (5H,m).

Step 2：The preparation of 4- hydroxyl -2- azabicyclos [4.1.0] heptane -2- carboxylic acid tert-butyl esters

Under nitrogen protection, to equipped with (the 2H)-benzyl carboxylate of 3- hydroxyls -3,4- dihydropyridine -1 (11.9g, Et is slowly added dropwise in (0 DEG C) in 1.2- dichloroethane solutions 51.01mmol)₂(75mL, 75.00mmol, 1mol/L ring are by Zn Alkane), CH₂I₂(6mL, 75.00mmol), this reactant mixture are warmed to room temperature and stir 6h naturally, after TLC detection reactions are completed, Saturation NaHCO is slowly added dropwise again₃Reaction is quenched in solution, and DCM extractions (50mL x4) are washed after merging organic phase with saturation NaCl, Anhydrous Na₂SO₄Fully dry, pass through chromatography (PE after vacuum evaporation：EA=3：1~1：1) purifying obtains 11.0g target chemical combination Thing, is white solid, yield 94.4%；

1H NMR(400MHz,CDCl₃)δ0.64-0.77(1H,m),0.84-0.97(1H,m),1.04-1.14(1H,m), 1.90-2.01 (3H, m), 2.86-2.93 (2H, m), 3.87 (1H, dd, J=13.2Hz, 4.8Hz), 3.97-4.05 (1H, m), 5.20(2H,s),7.30-7.40(5H,m).

Step 3：4- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -2- azabicyclos The preparation of [4.1.0] heptane -2- benzyl carboxylates

Under nitrogen protection, to equipped with 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine (B-H) (2.0g, 6.59mmol), 4- hydroxyl -2- azabicyclos [4.1.0] heptane -2- carboxylic acid tert-butyl esters (2.1g, 8.57mmol) and PPh₃DIAD (2.0g, 9.89mmol) is slowly added dropwise in (0 DEG C) in the 90mL THF solutions of (2.6g, 9.89mmol), and the reaction mixes 3h is stirred at room temperature in compound, states then up and water quenching is slowly added dropwise in reactant mixture goes out reaction, DCM extractions (50mL x 3), organic phase anhydrous Na₂SO₄Fully dry, pass through chromatography (DCM after vacuum evaporation：MeOH=50:1~20:1) purify It is white solid to 1.1g target compounds, yield 31.4%；EM (calculated value)：532.2；MS(ESI)m/e(M+1H)⁺： 533.3；

Step 4：1- (2- azabicyclos [4.1.0] heptane -4- bases) -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] The preparation of pyrimidine -4- amine

Under 2atm Hydrogen Vapor Pressures, (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] is phonetic by reaction system 4- Pyridine -1- bases) -2- azabicyclos [4.1.0] heptane -2- benzyl carboxylates (610mg, 1.15mmol), Pd-C (150mg, 10%W/W) It is stirred at room temperature in 20mL MeOH overnight, after reaction is completed, filters and remove Pd-C, pass through chromatography after vacuum evaporation (DCM：MeOH=40:1~10:1) purifying obtains 110mg target compounds, is white solid, yield 24.0%；EM (is calculated Value)：398.2；MS(ESI)m/e(M+1H)⁺：399.2；

Step 5：(4- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -2- azepines are double by 1- Ring [4.1.0] nonane -2- bases) propane -2- alkene -1- ketone preparation

Under nitrogen protection, to equipped with 1- (2- azabicyclos [4.1.0] heptane -4- bases) -3- (4- Phenoxyphenyls) - 1H- pyrazoles [3,4-d] pyrimidine -4- amine (310mg, 0.80mmol), TEA (162mg, 1.61mmol) 20mL DCM solution in (0 DEG C) acryloyl chloride (65mg, 0.72mmol) is slowly added dropwise, the reactant mixture is stirred at room temperature 3h, is then slowly added dropwise again Saturation NH₄Reaction is quenched in Cl, is washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄Fully dry, pass through after vacuum evaporation Chromatography (DCM：MeOH=20：1) purifying obtains 35mg target compounds, is white solid, yield 9.7%.

1H NMR(400MHz,d₆-DMSO)δ0.63-0.67(1H,m),1.05-1.11(1H,m),1.54-1.62(1H, M), 2.29-2.33 (1H, m), 2.61-2.71 (1H, m), 3.02 (1H, t, J=11.6Hz), 3.09-3.14 (1H, m), 4.33 (1H, dd, J=12.0Hz, 3.2Hz), 4.57-4.65 (1H, m), 5.80 (1H, dd, J=10.4Hz, 2.4Hz), 6.23 (1H, Dd, J=16.4Hz, 2.4Hz), 7.02 (1H, dd, J=16.4Hz, 10.4Hz), 7.12-7.21 (6H, m), 7.42-7.46 (2H, m), 7.67 (2H, d, J=8.8Hz), 8.24 (1H, s)

EM (calculated value)：452.2；MS(ESI)m/e(M+1H)⁺：453.2

Test example 1：External BTK suppresses Kinase activity assays

The external inhibitory activity experiments (test example 1 and test example 2) of BTK are surveyed by Shanghai Ruizhi Chemical Study Co., Ltd. It is fixed.

1：Test material：

BTK (Carna products, article No. 08-180, lot number 14CBS-0619D)；

Substrate polypeptide FAM-P2 (GL Biochem products, article No. 112394, lot number P100804-XZ112394)；Adenosine three Phosphoric acid (Sigma products, lot number A7699-1G, lot number 987-65-5)；

DMSO (Sigma products, article No. D2650, lot number 474382)；

Tetraacethyl diaminoethanes (Sigma products, article No. E5134, lot number 60-00-4)；

96- orifice plates (Corning products, article No. 3365, lot number 22008026)；

384- orifice plates (Corning products, article No. 3573, lot number 12608008)；

Positive control：Staurosporine (Sigma products, article No. S4400-1MG, lot number 046K4080)；

Detecting instrument：Caliper EZ Reader

2：Test principle：

The mobility detection technique (Mobility-Shift Assay) of microfluidic chip technology, the technology are electric by capillary The basic concept of swimming is applied in microfluidic environment, and the substrate for experiment is the polypeptide with fluorescence labeling, in reaction system In the presence of middle enzyme, substrate is changed into product, and also there occurs corresponding change, Mobility-Shift for the electric charge of its band Assay is exactly using the electrically charged difference of substrate and product, and the two is separated, and is detected respectively, testing result Expressed by conversion ratio.

3：Test method：

(1) testing sample is configured：50 times i.e. 25umol/L of reaction final concentration is diluted to 100%DMSO；

(2) dilute：25umol/L is initial concentration, then with 4 times of concentration dilutions, dilutes 10 concentration gradients；

(3) it is separately added into 100%DMSO in positive control and negative control hole；

(4) compound of prepare 10 concentration is diluted 10 times with 1 times of kinase buffer liquid respectively；Wherein kinase buffer liquid In comprising concentration be 50mmol/L, hydroxyethyl piperazine second thiosulfonic acid that pH is 7.5,0.01% Brij-35, 10mmol/L magnesium chloride, 2mmol/L dithiothreitol (DTT)；

(5) 2.5 times of enzyme solutions are prepared：Kinases is added into 1 times of kinase buffer liquid, forms 2.5 times of enzyme solutions；

(6) 2.5 times of substrate solution is prepared：The polypeptide of FAM marks and ATP are added into 1 times of kinase buffer liquid, form 2.5 Times substrate solution；

(7) enzyme solutions are added into 384 orifice plates：5 times of chemical combination of existing 5 μ l 10%DMSO dissolvings in 384 hole reaction plates Thing, 10 μ l 2.5 times of enzyme solutions are then added, be incubated 10 minutes at room temperature；

(8) substrate solution is added into 384 orifice plates：10 μ l 2.5 times of substrate solutions are added in 384 hole reaction plates；

(9) kinase reaction and termination：1h is incubated at 28 DEG C, then adds 25 μ l terminate liquid terminating reactions；Wherein in terminate liquid Be 100mmol/L comprising concentration, hydroxyethyl piperazine second thiosulfonic acid that pH is 7.5,0.015% Brij-35, 0.2% No. 3 surface reagents, 20mmol/L ethylenediamine tetra-acetic acid；

(10) Caliper reads reading and converting rate data on data Caliper；

(11) inhibiting rate calculates and conversion data is replicated from Caliper.

Conversion into inhibiting rate data, wherein max refers to the conversion ratio of DMSO controls, and min is no enzyme activity control Conversion ratio.

Percent inhibition=(max-conversion)/(max-min) * 100.

Test example 2：The measure of cell in vitro proliferation inhibition activity is carried out to compound from different cell lines

1：Test material：

RPMI1640 culture mediums (INVITROGEN products, article No. 61870-127)；

Hyclone (GIBCO products, article No. 10099-141)；

Trypsase (GIBCO products, article No. 25200-072)；

Luminescence method cell viability detection kit (Promega products, article No. G7572)；

96 porocyte culture plates, black (Costar products, article No. 3904)；

Bottom white pad pasting (PE products, article No. 6005199)；

DMSO (Sigma products, article No. D2650)；

Paclitaxel (domestic vendors)

Cell line：DOHH2、WSU-DLCL2.

2：Method for preparation of drug：

Medicine to be measured is configured to 10mmol/L solution using DMSO, standby after concussion dissolving.

3：Cell injuring model method：

Two plants of cells DOHH2, the WSU-DLCL2 chosen use the RPMI1640 nutrient solutions containing 10% hyclone to be trained Support, at 37 DEG C, 5% CO₂Wet condition under be incubated.

4：Experimental method：

First day：Plating cells

(1) under cell is digested from cell culture using pancreatin, (10% hyclone is contained using complete medium RPMI1640 nutrient solutions) be resuspended after determine cell density；

(2) cell is diluted to the cell solution of standard density；

(3) by the cell solution after adjustment density, (point of addition is 2 using in every 90 microlitres of 96 hole cell experiment plates of addition in hole ~12 row, the cell free broth of the 1st row supplement respective volume)；

(4) Tissue Culture Plate completed is put into incubator, at 37 DEG C, 5% CO₂24h is incubated under wet condition.

Second day：Add compound

(1) according to the reference compound of experiment pattern 200 times of concentration of preparation and compound solution to be tested, (compound is tested Initial concentration is 50umol/L, and gradient dilution, extension rate are carried out using DMSO using 200 times of concentration 10mmol/L as initial concentration For 3 times, 9 concentration points；Reference compound test initial concentration is 1umol/L, dense to originate using 200 times of concentration 0.2mmol/L Degree carries out gradient dilution using DMSO, and extension rate is 3 times, 9 concentration points)；

(2) take 7 microlitres of compound solutions to be added in 133 microlitres of complete mediums to be diluted, shake up 10 minutes (DMSO Final concentration 5%)；

(3) taking the compound solution after 10 microlitres of dilutions to add in the previous day ready Tissue Culture Plate, (DMSO is dense eventually Degree is 0.5%)；

(4) Tissue Culture Plate for adding compound places back in incubator, at 37 DEG C, 5% CO₂Wet condition under be incubated 72h。

5th day：As a result detect

(1) detection reagent is positioned over room temperature in 30 minutes before experiment and is balanced；

(2) rocker 10 minutes under the conditions of the every hole addition of Tissue Culture Plate 30 microlitres of detection reagents, lucifuges, inducing cell splits Solution；

Tissue Culture Plate is incubated 2 minutes with stabilized illumination signal at room temperature after (3) 10 minutes；

(4) brassboard lower, planar sticks white pad pasting；

(5) use Envision read plates, during read plate the time be set as 0.5 second per hole.

The calculation formula of cell inhibitory rate：

% inhibiting rates=(maximum signal level-compound signal value)/(maximum signal level-minimum signal value) × 100.

The signal value of maximum is obtained from dmso treatment 72h cell；From single culture medium (cell number zero) Obtain minimum signal value.

Calculated using XLfit softwares

Part of compounds is to BTK kinase inhibiting activities and cell in vitro proliferation inhibition activity result：

As can be seen from the table, embodiment 2,3,4,5,8,9 all has obvious inhibitory action to BTK activity, with Positive control is roughly the same for Buddhist nun's effect according to Shandong, illustrates the suppression that piperidine ring is replaced with to the compounds such as some loop coils or bridged ring Effect does not have much affect substantially, but still can show potent BTK kinase inhibiting activities.But on a cellular level, Two COMPOUNDS EXAMPLEs 4 and embodiment 8 tested show the cell inhibitory effect more more potent than positive control Activity, especially inhibitory activity of the embodiment 4 on DOHH2 cells improve similar 8 times.Therefore, designed by the present invention Loop coil or endocyclic compound can be used as BTK inhibitor, have the application prospect of wide anti-malignant tumor.

Claims

1. a kind of bruton's tyrosine kinase inhibitor with loop coil or caged scaffold, it is characterised in that including formula (I)：

Wherein Y is selected from following groups：

Wherein, the N-terminal of Y group is connected with carbonyl, and C-terminal is connected with the N in pyrazolopyrimidine ring, and R1, R2 can be independently selected from Hydrogen atom, fluorine atom, bromine atoms, iodine atom, amino, cyano group, sulfydryl, C₁~C₅Straight or branched alkyl；One on the alkyl Individual or multiple hydrogen atoms can be substituted by one or more halogen, amino, cyano group, hydroxyl, sulfenyl；

Wherein, m₁、m₂It is C₁~C₂Alkyl；

n₁、n₂It is C₁~C₃Alkyl；

G is O.

2. a kind of bruton's tyrosine kinase inhibitor with loop coil or caged scaffold according to claim 1, it is special Sign is, selected from following structure：

3. a kind of bruton's tyrosine kinase inhibitor with loop coil or caged scaffold according to claim 2, it is special Sign is, selected from following structure：

。

4. 1- (3- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- as claimed in claim 3 Base) -1- oxa- -8- azaspiros [4.5] decane -8- bases) propyl- 2- alkene -1- ketone preparation method, it is characterised in that including following Step：

(1) hydroxy piperidine -1- t-butyl formates of 4- pi-allyls -4 are prepared

Under nitrogen protection, allyl bromide, bromoallylene is added dropwise into the dry diethyl ether solution 20mL containing 3.2g magnesium powders, reaction continues to drip after triggering Adding allyl bromide, bromoallylene, rate of addition is controlled in maintenance reaction reflux state, adds allyl bromide, bromoallylene 13.3g altogether twice, be subsequently cooled to- 15 DEG C, the THF solution 100mL of the tertbutyloxycarbonyl -4- piperidones of N- containing 10.0g is added, stirring reaction 5h, is subsequently cooled to 0 DEG C, add saturation NH₄Cl extracts reaction of going out, and separates organic phase, extracts, and dries, and the hydroxyl piperazine of 4- pi-allyls -4 is obtained after vacuum evaporation Pyridine -1- t-butyl formates；

(2) 3- hydroxyls -1- oxa- -8- azaspiros [4.5] decane -8- t-butyl formates are prepared

Mixing to the 100mL tert-butyl alcohols and 40mL water containing the hydroxy piperidine -1- t-butyl formates of 11.3g 4- pi-allyls -4 is molten 11.1g NaIO are added in liquid₄, 50 DEG C of stirring reaction 0.5h, then add Na containing 9.8g₂S₂O₅Aqueous solution 30mL, 50 DEG C are anti- 7h is answered, 48h is stirred at room temperature, finally plus water extracts reaction of going out, and extracts, dries, 3- hydroxyl -1- oxa- -8- azepines are obtained after vacuum evaporation Loop coil [4.5] decane -8- t-butyl formates；

(3) 3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxa- -8- azepines are prepared Loop coil [4.5] decane -8- t-butyl formates

Nitrogen protection under, to 3- containing 1.2g (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine, 1.6g 3- hydroxyls - 1- oxa- -8- azaspiros [4.5] decane -8- t-butyl formates and 2.1g PPh₃THF solution 20mL in, be added dropwise at 0 DEG C 1.6g DIAD, stirring reaction 3h, adds water and reaction is quenched at room temperature, extraction, fully dry, be evaporated in vacuo 3- (4- amidos- 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxa- -8- azaspiros [4.5] decane -8- formic acid uncles Butyl ester；

(4) 3- (4- Phenoxyphenyls) -1- (1- oxa- -8- azaspiros [4.5] decane -3- bases) -1H- pyrazoles [3,4-d] is prepared Pyrimidine -4- amine

At room temperature, to containing 1.5g 3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- 5mL TFA are added dropwise in the DCM solution 40mL of oxa- -8- azaspiros [4.5] decane -8- t-butyl formates, stirring is anti-at room temperature 2h is answered, NaHCO is then added dropwise₃To pH be more than 7, separate organic phase, extract, dry, be evaporated in vacuo 3- (4- Phenoxyphenyls)- 1- (1- oxa- -8- azaspiros [4.5] decane -3- bases) -1H- pyrazoles [3,4-d] pyrimidine -4- amine；

(5) 1- (3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1-yl) -1- oxa- -8- nitrogen is prepared Miscellaneous spiral shell [4.5] decane -8-yl) propane -2- alkene -1- ketone

Under nitrogen protection, to containing 751mg 3- (4- Phenoxyphenyls) -1- (1- oxa- -8- azaspiros [4.5] decane -3- Base) -1H- pyrazoles [3,4-d] pyrimidine -4- amine, 344mg TEA DCM solution 20mL in, at 0 DEG C be added dropwise 139mg acryloyl chlorides, Stirring reaction 3h at room temperature, saturation NH is then added dropwise again₄Reaction is quenched in Cl, is washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄Fully dry, 1- (3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- are obtained after vacuum evaporation Yl) -1- oxa-s -8- azaspiros [4.5] decane -8-yl) propane -2- alkene -1- ketone.

5. 1- (3- (4- amino -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- as claimed in claim 3 Base) -1- oxa- -7- azaspiros [4.4] nonyl- 7- yls) propyl- 2- alkene -1- ketone preparation method, it is characterised in that including following step Suddenly：

(1) 3- pi-allyl -3- hydroxypyrrole -1- carboxylic acid tert-butyl esters are prepared

Under nitrogen protection, to the THF solution 20mL containing 5.0g 1- t-butoxycarbonyl -3- pyrrolidones, 8.2g allyl bromide, bromoallylenes In, 28mL saturations NH is added at 0 DEG C₄Cl, 3.8g zinc powders are then added, keep reaction temperature to be no more than 40 in adition process DEG C, reaction 12h is then stirred at room temperature, 2mol/L H are finally added dropwise₂SO₄Reaction is quenched, EA extractions, is used after merging organic phase Saturation NaCl is washed, anhydrous Na₂SO₄Dry, 3- pi-allyl -3- hydroxypyrrole -1- carboxylic acid tert-butyl esters are obtained after vacuum evaporation；

(2) 3- (2,3- dibromopropyls) -3- hydroxypyrrole -1- carboxylic acid tert-butyl esters are prepared

Nitrogen protection under, into the DCM solution 10mL containing 1.0g 3- pi-allyl -3- hydroxypyrrole -1- carboxylic acid tert-butyl esters in - 703mg Br are added dropwise at 15 DEG C₂, 0.5h is stirred at room temperature again afterwards, is eventually adding saturation Na₂S₂O₃Extract reaction of going out, DCM extractions, close And saturation NaHCO is used after organic phase₃Washing, anhydrous Na₂SO₄Dry, 3- (2,3- dibromopropyl) -3- hydroxyls are obtained after vacuum evaporation Pyrroles's -1- carboxylic acid tert-butyl esters；

(3) bromo- 1- carbonyls -7- azaspiros [4.4] nonane -7- carboxylic acid tert-butyl esters of 3- are prepared

Under nitrogen protection, to the 10mL MeOH solution containing 3- (2,3- dibromopropyl) -3- hydroxypyrrole -1- carboxylic acid tert-butyl esters In add 857mg K at room temperature₂CO₃, it is stirred overnight at room temperature again afterwards, then adds water and extract reaction of going out, DCM extractions, be associated with Anhydrous Na is used after machine phase₂SO₄Dry, bromo- 1- carbonyls -7- azaspiros [4.4] nonane -7- carboxylic acid tert-butyl esters of 3- are obtained after vacuum evaporation；

(4) 3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxo -7- azepines are prepared Spiral shell [4.4] nonane -7- carboxylic acid tert-butyl esters

By 600mg 3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -4- amine, the bromo- 1- carbonyls -7- azepines of 600mg 3- Spiral shell [4.4] nonane -7- carboxylic acid tert-butyl esters, 2.0g Cs₂CO₃It is stirred overnight in 85 DEG C in 20mL DMF, is cooled down after the completion of reaction To room temperature, then add water and extract reaction of going out, EA extractions, anhydrous Na₂SO₄Dry, 3- (4- amidos -3- (4- benzene is obtained after vacuum evaporation Phenyl) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxo -7- azaspiros [4.4] nonane -7- carboxylic acid tert-butyl esters；

(5) 3- (4- Phenoxyphenyls) -1- (1- oxo -7- azaspiros [4.4] nonane -3- bases) -1H- pyrazoles [3,4-d] is prepared Pyrimidine -4- amine

At room temperature, to 1.0g 3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxos - 60mL DCM solution is added in 7- azaspiros [4.4] nonane -7- carboxylic acid tert-butyl esters is completely dissolved it, then adds 200mL Saturation HCl EA solution, the reactant mixture are stirred at room temperature overnight, and are evaporated in vacuo after the completion of reaction to dry, then use 300mL DCM dissolves, and this DCM organic phase uses saturation NaHCO respectively₃With saturated common salt water washing, organic phase anhydrous Na₂SO₄It is fully dry It is dry, after vacuum evaporation 3- (4- Phenoxyphenyls) -1- (1- oxo -7- azaspiros [4.4] nonane -3- bases) -1H- pyrazoles [3, 4-d] pyrimidine -4- amine；

(6) 1- (3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) -1- oxo -7- nitrogen is prepared Miscellaneous spiral shell [4.4] nonane -7- bases) propane -2- alkene -1- ketone

Under nitrogen protection, to containing 300mg 3- (4- Phenoxyphenyls) -1- (1- oxo -7- azaspiros [4.4] nonane -3- Base) -1H- pyrazoles [3,4-d] pyrimidine -4- amine, 141mg TEA DCM solution 20mL in, at 0 DEG C be added dropwise 64mg acryloyl chlorides, room The lower stirring reaction 3h of temperature, is then added dropwise saturation NH4Cl and reaction is quenched, washed after separating organic phase with saturation NaCl, anhydrous Na₂SO₄ Fully dry, after vacuum evaporation 1- (3- (4- amidos -3- (4- Phenoxyphenyls) -1H- pyrazoles [3,4-d] pyrimidine -1- bases) - 1- oxo -7- azaspiros [4.4] nonane -7- bases) propane -2- alkene -1- ketone.

6. the bruton's tyrosine kinase inhibitor with loop coil or caged scaffold described in claim any one of 1-3 is being made Purposes on standby inflammatory conditions and/or malignant tumor medicine.