CN109970740A

CN109970740A - 4- amidino-pyridine and azepine ring derivatives and its preparation method and application

Info

Publication number: CN109970740A
Application number: CN201711448020.5A
Authority: CN
Inventors: 陈俐娟; 龙超峰; 陈小新; 刘卓伟; 钱志勇; 杨金亮; 向明礼
Original assignee: Guangdong Zhongsheng Pharmaceutical Co Ltd
Current assignee: Guangdong Zhongsheng Pharmaceutical Co Ltd
Priority date: 2017-12-27
Filing date: 2017-12-27
Publication date: 2019-07-05

Abstract

The invention belongs to biochemical fields, and in particular to 4- amidino-pyridine and azepine ring derivatives and its preparation method and application.The present invention provides a kind of 4- amidino-pyridine and azepine ring derivatives, and structural formula is as shown in formula Ⅹ.In addition, the present invention also provides the preparation method of the analog derivative and purposes.4- amidino-pyridine provided by the invention and azepine ring derivatives have the function of inhibiting BTK.

Description

4- amidino-pyridine and azepine ring derivatives and its preparation method and application

Technical field

The invention belongs to chemicals technical fields, in particular to 4- amidino-pyridine and azepine ring derivatives and its preparation Method and purposes.

Background technique

Protein kinase is made of a series of relevant enzyme in structures, is mainly responsible for the control of intracellular signal transduction process. In general, phosphinylidyne group-transfer of the protein kinase by influence from ribonucleoside triphosphote to the protein receptor for participating in signal transduction path, and Mediate intracellular signal.These phosphorylation events play the role of modulation or adjust the molecular switch of target protein biological function.Very much Disease is all related with the abnormal cell reaction caused by above-mentioned protein kinase mediated event.

BTK (Bruton ' s tyrosine kinase) i.e. bruton's tyrosine protein kinase, is non-receptor tyrosine kinase The member of enzyme Tec family, is almost expressed by hematopoietic cell, is the crucial composition portion of B-cell receptor (BCR) signal path Point, while also mediation is played in proinflammatory signal communication.The BCR on B cell surface and its antigen binding, are lured by antigen BCR aggregation is led, activates the Src family molecule such as the Lyn and Fyn in downstream, the Src family phosphorylation of activation is in Tyr551 and Tyr223 The BTK in site.The BTK of activation can by further phosphorylation PLCg2 increase intracellular calcium concentration, MAPKs, NF-kB and The activation etc. of AKT signal path, may stimulate the phosphorylation in these sites in phosphatase, thus controlling gene and cell because The expression of son, influences survival, proliferation and the differentiation of B cell.BTK mutation can destroy BTK function, and patient's body is caused to lack antibody With mature periphery bone-marrow-derived lymphocyte.BTK activates Fc γ R signal by immunocomplex in Functions of Bone Marrow Cells.It is thin in macrophage In born of the same parents, BTK inhibit can block Fc γ RIII induce INF- α, IL-1 β and IL-6 product, by NF-kB activate block BCR according to Bad B cell proliferation, this provides strong evidence in the treatment of autoimmune disease for BTK, including rheumatoid arthritis and is System property ruthless sore of erythema etc..In addition, it is the target spot for treating lymthoma related disease that BTK has been confirmed in many ways.

Traditional most of tyrosine kinase inhibitor (TKI) is with reversible active force (hydrogen bond, Van der Waals force and hydrophobic work Firmly etc.) highly conserved ATP binding domain (occupying " switch " part) is incorporated into prevent or reduce the phosphorus of tyrosine kinase Acidification, this weaker and invertibity active force basis are that this kind of drugs bring 3 very important disadvantages: selectivity is not It is enough；Drug effect is not strong enough and lasting；Easily cause drug resistance.In recent years, covalent TKI causes the favor of scholar, in particular for Antitumor, antibacterial and antiviral drugs.The ratified till now through the breakthrough drug channel of FDA from the aspirin before 100 years Two new drugs replace Buddhist nun (Ibrutinib) according to Shandong, and report irreversible covalently small point of more antitumor resistance mechanism at present Sub- compound shows advantage of the covalent compound in treatment chronic disease.Wherein, irreversibility TKI is usually with reversible Property TKI skeleton structure be prototype, in place connect an electrophilic functional group, such as α, beta-unsaturated aldehyde/ketone structure etc., With the cysteine residues (the nucleophilic structure of electron rich) near ATP binding domain electrophilic reaction can occur for this electrophilic functional group Form covalent bond.Compared with invertibity TKI, irreversible TKI has unique advantage.Firstly, irreversible TKI is permanently to inactivate Mode plays a role, and it is strong and lasting to enable drug effect.Secondly, the combination of irreversible TKI and ATP, kinases and there is no competition Property, so that reducing the generation that drug resistance is alleviated or avoided a possibility that kinase mutant.In addition, the selectivity of irreversible TKI It is very high, because of the electrophilic functional group property of can choose that is divided into minor structure in conjunction with the sulfydryl on cysteine residues, especially It is that can relatively easily improve selectivity for the high kinase members of homology, and avoid bring side effect of missing the target.It can not Ignore, since the presence of electrophilic group can make drug molecule and albumen extended durations of action, if drug molecule selectivity is not Height, has also carried out covalent bond with non-target albumen, then can greatly increase the risk of not expected toxic reaction.Therefore, When setting covalent TKI for disease treatment, it is necessary to which the selectivity for improving targeted drug has balanced drug and protein amino acid residues Binding ability, avoid drug from missing the target brought adverse reaction.

BTK is expressed in hematopoietic cell, mediates BCR and Fr γ R signal access, and inhibitor can reduce by antigen, be immunized again Close inflammatory reaction caused by object and pathogen.Currently, animal case of the BTK inhibitor for treating rheumatoid arthritis is reported It is more, including ATP competitive inhibitor and covalency inhibitor.Wherein, the unique marketed drug of BTK replaces Buddhist nun (ibrutinib) according to Shandong, It is also at arthritic preclinical study at present, effect is more significant.Since arthritis is not a kind of disease of threat to life, Use safety exquisite when drug therapy；Meanwhile arthritic is difficult to eradicate, and long-term administration is needed to treat, therefore develop efficient , the newtype drug of long half time, the compliance of patient can be improved, improve the treatment and quality of life of patient.Buddhist nun is replaced according to Shandong, is made For covalency inhibitor, it can efficiently inhibit BTK, but its selectivity is very poor, there are good suppressions to the kinases of many cancer targets Production is used, such as EGFR family kinase and Src family kinase.Many is had also discovered during clinical application not for Buddhist nun according to Shandong Good reaction, such as Toxicity of Kidney, diarrhea, bleeding and bone marrow suppression；Simultaneously according to Shandong for Buddhist nun to the inhibition of hERG potassium-channel compared with (IC by force₅₀=0.97 μM), long-time service is possible to will appear cardiac risk.In addition, it is not high for the bioavilability of Buddhist nun according to Shandong, it gives Medicine is difficult, if undoubtedly also will increase the risk that various ill-effects occur by increasing dosage come treatment of arthritis.According to Shandong It is as follows for Buddhist nun's structural formula:

Currently, developing selective stronger, the higher covalent compound of activity to be effectively very urgent.

Summary of the invention

First technical problem to be solved by this invention is to provide 4- amidino-pyridine and azepine ring derivatives, structural formula As shown in Ⅹ:

Wherein, X is C or N；R₂ForSubstituted or unsubstituted nitrogenous 5 membered heterocycloalkyl orR₃ForOr substituted or unsubstituted nitrogenous 6 membered heterocycloalkyl；Nitrogenous 5 membered heterocycloalkyl, nitrogenous 6 circle heterocyclic ring alkane Substituent group on basic ring is independently-H, C1~C8 alkyl, halogen or-CN；R₄, substituent group on nitrogenous 5 membered heterocycloalkyl nitrogen, Substituent group on nitrogenous 6 membered heterocycloalkyl nitrogen is independentlyR₅、R₆Be independently C2~ C8 alkenyl, C2~C8 alkynyl,R₇、R₈It is independently-H or C1~C8 alkyl；M is 1~4； N is 0~4；Z, p is 0~4；

R₁ForThe cycloalkenyl of C3~C8,Or it is substituted or unsubstituted 5~10 unit's heteroaryls；The hetero atom of 5~10 unit's heteroaryl is that hetero atom is N, O, S, and hetero atom number is 1~3； The substituent group of 5~10 unit's heteroaryl is-H, halogen or C1~C8 alkyl；R₉~R₁₃It is independently-H, halogen, C1~C8 alkane Base, C1~C8 alkoxy ,-CF₃、-OCF₃、Phenyl, phenoxy group, benzyloxy, C1~C8 oxygen carbonyl,C is 0~4；Or R₁₀And R₁₁Combination forms 3~8 circle heterocyclic rings Alkyl or 3~8 membered unsaturated heterocycles；3~8 membered heterocycloalkyl, 3~8 membered unsaturated heterocycles hetero atom be N, O, S, it is miscellaneous Atom number is 1~3.

As a preferred solution of the present invention, in above-mentioned 4- amidino-pyridine and azepine ring derivatives, X is C or N；R₂ForSubstituted or unsubstituted nitrogenous 5 membered heterocycloalkyl orR₃ForOr replaces or do not take Nitrogenous 6 membered heterocycloalkyl in generation；Substituent group in nitrogenous 5 membered heterocycloalkyl, nitrogenous 6 membered heterocycloalkyl ring is independently- H, C1~C6 alkyl, halogen or-CN；R₄, substituent group on nitrogenous 5 membered heterocycloalkyl nitrogen, taking on nitrogenous 6 membered heterocycloalkyl nitrogen Dai Ji is independentlyR₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl,R₇、R₈It is independently-H or C1~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

R₁ForThe cycloalkenyl of C3~C6,Or it is substituted or unsubstituted 5~8 unit's heteroaryls；The hetero atom of 5~8 unit's heteroaryl is that hetero atom is N, O, S, and hetero atom number is 1~3；Institute The substituent group for stating 5~8 unit's heteroaryls is-H, halogen or C1~C6 alkyl；R₉~R₁₃Be independently-H, halogen, C1~C6 alkyl, C1~C6 alkoxy ,-CF₃、-OCF₃、Phenyl, phenoxy group, benzyloxy, C1~C6 oxygen carbonyl,C is 0~4；Or R₁₀And R₁₁Combination forms 3~6 circle heterocyclic rings Alkyl or 3~6 membered unsaturated heterocycles；3~6 membered heterocycloalkyl, 3~6 membered unsaturated heterocycles hetero atom be N, O, S, it is miscellaneous Atom number is 1~3.

Preferably, X is C or N；R₂ForR₃ForR₁₅~R₁₇、R₁₉~R₂₁、R₂₃~ R₂₆、R₂₈~R₃₁、R₃₃~R₃₆It is independently-H, C1~C6 alkyl, halogen or-CN；R₄、R₁₄、R₁₈、R₂₂、R₂₇、R₃₂It is independentlyR₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl, R₇、R₈It is independently-H or C1~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

It is further preferred that X is C or N；R₂For R₃ForR₁₅~R₁₇、R₁₉~R₂₁、R₂₃ ~R₂₆、R₂₈~R₃₁、R₃₃~R₃₆It is independently-H, C1~C4 alkyl, halogen or-CN；R₄、R₁₄、R₁₈、R₂₂、R₂₇、R₃₂It is independent ForR₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~3；N is 0~3；Z, p is 0~3；

R₁ForThe cycloalkenyl of C3~C6,Or it is substituted or unsubstituted 5~6 unit's heteroaryls；The hetero atom of 5~6 unit's heteroaryl is that hetero atom is N, O, S, and hetero atom number is 1~2；Institute The substituent group for stating 5~6 unit's heteroaryls is-H, halogen or C1~C4 alkyl；R₉~R₁₃Be independently-H, halogen, C1~C4 alkyl, C1~C4 alkoxy ,-CF₃、-OCF₃、Phenyl, phenoxy group, benzyloxy, C1~C4 oxygen carbonyl,C is 0~3；Or R₁₀And R₁₁Combination forms 3~6 circle heterocyclic rings Alkyl or 3~6 membered unsaturated heterocycles；3~6 membered heterocycloalkyl, 3~6 membered unsaturated heterocycles hetero atom be N, O, S, it is miscellaneous Atom number is 1~2.

Still further preferably, X is C or N；R₂For R₃ForR₁₅~R₁₇、R₁₉~R₂₁、R₂₃ ~R₂₆、R₂₈~R₃₁、R₃₃~R₃₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₄、R₁₄、R₁₈、R₂₂、R₂₇、R₃₂ It is independentlyR₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Still more preferably, X is C or N；R₂For R₃ForR₁₅~R₁₇、R₁₉~R₂₁、 R₂₃~R₂₆、R₂₈~R₃₁、R₃₃~R₃₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₄、R₁₄、R₁₈、R₂₂、R₂₇、 R₃₂It is independentlyR₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

R₁ForThe cycloalkenyl of C3~C6,Or it is substituted or unsubstituted 6 membered nitrogen-containing heteroaryl bases；The substituent group of the 6 membered nitrogen-containing heteroaryl base is-H, halogen or C1~C4 alkyl；R₉~R₁₃It is independent For-H ,-F ,-Cl ,-Br, C1~C4 alkyl, C1~C4 alkoxy ,-CF₃、-OCF₃、Phenyl, phenoxy group, Benzyloxy, C1~C4 oxygen carbonyl,C is 0~2；Or R₁₀With R₁₁Combination forms 3~6 membered heterocycloalkyls or 3~6 membered unsaturated heterocycles；The hetero atom of 3~6 membered heterocycloalkyl is O, miscellaneous original Sub- number is 1~2；The hetero atom of 3~6 membered unsaturated heterocycle is N, and hetero atom number is 1~2.

Most preferably, X is C or N；R₂ForR₃ForR₁₅~R₁₇、R₁₉~R₂₁、R₂₃~ R₂₆、R₂₈~R₃₁、R₃₃~R₃₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₄、R₁₄、R₁₈、R₂₂、R₂₇、R₃₂Solely It is vertical to beR₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1 or 2；N is 1 or 2；Z is 0；P is 1 Or 2；

R₁For R₉~R₁₃It is independently-H ,-F ,-Cl ,-Br, C1~C4 alkyl, C1~C4 alcoxyl Base ,-CF₃、-OCF₃、Phenyl, phenoxy group, benzyloxy, C1~C4 oxygen carbonyl, C is 0 or 1.

As a preferred solution of the present invention, above-mentioned 4- amidino-pyridine and azepine ring derivatives, work as R₂ForM is 1, R₃ForWhen X is C, structural formula is as shown in formula I:

Wherein, R₄ForR₅、R₆Be independently C2~C8 alkenyl, C2~C8 alkynyl,R₇、R₈It is independently-H or C1~C8 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

Preferably, R₄ForR₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl,R₇、R₈It is independently-H or C1~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

It is further preferred that R₄ForR₅、R₆Be independently C2~C4 alkenyl, C2~ C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~3；N is 0~3；z,p It is 0~3；

Still further preferably, R₄ForR₅、R₆It is independently C2~C4 alkenyl, C2 ~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2； Z, p is 0~2；

Still more preferably, R₄ForR₅、R₆It is independently C2~C4 alkenyl, C2 ~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2； Z, p is 0~2；

Most preferably, R₄ForR₅、R₆It is independently C2~C4 alkenyl, C2~C4 alkynes Base,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1 or 2；N is 1 or 2；Z is 0；p It is 1 or 2；

As a preferred solution of the present invention, above-mentioned 4- amidino-pyridine and azepine ring derivatives, work as R₂ForM is 1, R₃ForWhen X is C, structural formula is as shown in formula II:

Wherein, R₂₈~R₃₁It is independently-H, C1~C6 alkyl, halogen or-CN；R₂₇ForR₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl,R₇、R₈It is independently-H or C1~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

Preferably, R₂₈~R₃₁It is independently-H, C1~C4 alkyl, halogen or-CN；R₂₇For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independent For-H or C1~C4 alkyl；M is 1~3；N is 0~3；Z, p is 0~3；

Further preferably, R₂₈~R₃₁It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₇For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Still more preferably, R₂₈~R₃₁It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₇For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Most preferably, R₂₈~R₃₁It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₇For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1 or 2；N is 1 or 2；Z is 0；P is 1 Or 2；

As a preferred solution of the present invention, above-mentioned 4- amidino-pyridine and azepine ring derivatives, work as R₂ForM is 1, R₃ForWhen X is C, structural formula is as shown in formula III:

Wherein, R₂₃~R₂₆It is independently-H, C1~C6 alkyl, halogen or-CN；R₂₂ForR₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl,R₇、R₈It is independently-H or C1~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

Preferably, R₂₃~R₂₆It is independently-H, C1~C4 alkyl, halogen or-CN；R₂₂For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independent For-H or C1~C4 alkyl；M is 1~3；N is 0~3；Z, p is 0~3；

Further preferably, R₂₃~R₂₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₂For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Still more preferably, R₂₃~R₂₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₂For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Most preferably, R₂₃~R₂₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₂For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1 or 2；N is 1 or 2；Z is 0；P is 1 Or 2；

As a preferred solution of the present invention, above-mentioned 4- amidino-pyridine and azepine ring derivatives, work as R₂ForM is 2, R₃ForWhen X is C, structural formula is as shown in formula VII:

As a preferred solution of the present invention, above-mentioned 4- amidino-pyridine and azepine ring derivatives, work as R₂ForX When for C, structural formula is as shown in formula IV:

Wherein, R₁₅~R₁₇It is independently-H, C1~C6 alkyl, halogen or-CN；R₁₄ForR₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl,R₇、R₈It is independently-H or C1~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

Preferably, R₁₅~R₁₇It is independently-H, C1~C4 alkyl, halogen or-CN；R₁₄For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independent For-H or C1~C4 alkyl；M is 1~3；N is 0~3；Z, p is 0~3；

Further preferably, R₁₅~R₁₇It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₁₄For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Still more preferably, R₁₅~R₁₇It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₁₄For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Most preferably, R₁₅~R₁₇It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₁₄It is independently R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1 or 2；N is 1 or 2；Z is 0；P is 1 Or 2；

As a preferred solution of the present invention, above-mentioned 4- amidino-pyridine and azepine ring derivatives, work as R₂ForX When for N, structural formula is as shown in formula VIII:

As a preferred solution of the present invention, above-mentioned 4- amidino-pyridine and azepine ring derivatives, work as R₂ForM is 1, R₃ForWhen X is N, structural formula is as shown in formula VI:

Above-mentioned 4- amidino-pyridine and azepine ring derivatives, structural formula are as follows:

Second technical problem to be solved by this invention is to provide the system of above-mentioned 4- amidino-pyridine and azepine ring derivatives Preparation Method.Synthetic route is as follows:

Work as R₂ForM is 1, R₃ForX is C, generalformulaⅰcompound the preparation method is as follows:

Step a: being added raw material 1 (1.0eq) into reaction flask, and enough THF or DMF is added as solvent, stirs in room temperature, Raw material NIS (1.1eq) is added portionwise under nitrogen protection, reaction solution is become to clarify dark-brown by suspension shape after reacting 1h at room temperature Liquid, the reaction was continued about 5h or so are spin-dried for reaction solution, add water to be dispersed solid and dissolve water-solubility impurity, be collected by filtration Filter cake, massive laundering wash filter cake, and drying filter cake obtains intermediate 2；

Step b: intermediate 2 (1.0eq), nitrobenzyl bromine (1.0eq) and K are added into reaction flask₂CO₃(1.5eq), adds Enter suitable DMF as solvent, stirred in room temperature, solution gradually becomes suspension by clarification, and TLC monitoring reaction terminates, and adds water Quenching reaction filters and washs filter cake with massive laundering, collects filter cake and dries to obtain intermediate 3；

Step c: intermediate 3, appropriate Isosorbide-5-Nitrae-dioxane and 25% ammonium hydroxide being added into autoclave, overnight in 98 DEG C Reaction；Next day is water-cooled reaction kettle, deflates, and takes out reaction solution and by it and is spin-dried for, water-dispersible solid, is collected by filtration solid Body, is repeatedly washed with water filter cake, and drying filter cake obtains intermediate 4；

Step d: intermediate 4 (1.0eq), raw material are added into there-necked flask(1.1eq)、PdCl2(dppf) Isosorbide-5-Nitrae-dioxane/H2O (9/1, v/ is added after N2 more times are replaced the air of there-necked flask in (0.1eq) and K2CO3 (3.0eq) v)；It is heated to reflux stirring 4h, insoluble matter is filtered to remove with diatomite, collects filtrate, be spin-dried for filtrate, extracted with EtOAc and water Filtrate is taken, collects oil reservoir, then with saturated common salt water washing oil reservoir 3 times, the dry oil reservoir of anhydrous Na 2SO4 is spin-dried for oil reservoir and uses silica gel Column purification collects intermediate 5；

Step e: intermediate 5 (1.0eq), NH4COOH (3.0eq) are added into reaction flask, appropriate MeOH makees solvent, in N2 Protection is lower to be added 10%Pd/C (10% mass of raw material 5), is heated to reflux 2h, and TLC monitoring reaction terminates；Diatomite is filtered to remove Insoluble matter collects filtrate, is spin-dried for filtrate, extracts filtrate with EtOAc and water, collection oil reservoir, then with saturated common salt water washing Oil reservoir 3 times, the dry oil reservoir of anhydrous Na 2SO4 is spin-dried for oil reservoir up to intermediate 6；

Step f: intermediate 6 (1.0eq) being added into reaction flask and appropriate super dry DMF makees solvent, and DIEA is slowly added dropwise (5.0eq), solution becomes clarification by muddiness, then is slowly added dropwise(for acyl chlorides or sulfonic acid chloride) (1.5eq), reaction is overnight；Add water Quenching reaction liquid, DCM extraction, saturated salt solution 3 times washings, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain mesh Mark product I.

Work as R₂ForM is 1, R₃ForX be C when, Compounds of formula II the preparation method is as follows:

Step g: being added raw material 7 (1.0eq) into reaction flask, and enough DCM are added as solvent, stir in ice bath, Et is added₃It is slowly added dropwise after N (1.5eq) methylsufonyl chloride (1.2eq), solution is changed from clarification shape to suspension, continues to stir (sulphur is aobvious), fully reacting are reacted in TLC monitoring after mixing 3h；It is filtered to remove insoluble matter, collects filtrate, by extracting repeatedly with saturation food Salt water washing filtrate, the dry oil reservoir of anhydrous sodium sulfate, is spin-dried for oil reservoir and obtains oil product intermediate 8, without purifying；

Step h: being added intermediate 8 (1.0eq) into reaction flask, and enough ACE are added as solvent, stir in room temperature, It is slowly added to LiBr (3.0eq), is gradually heated to reflux state, overnight；Reaction solution is cooled to room temperature, adds water quenching reaction Liquid is extracted after being spin-dried for reaction solution with EtOAc and water, collects oil reservoir, then with saturated common salt water washing oil reservoir 3 times, anhydrous Na₂SO₄Dry oil reservoir is spin-dried for oil reservoir up to oil product intermediate 9, without purifying；

Step i: intermediate 2 (1.0eq), intermediate 9 (1.0eq) and K are added into reaction flask₂CO₃(1.5eq) is added suitable It measures DMF and is used as solvent, stir in 60 DEG C, TLC monitoring reaction terminates, and add water and EtOAc to carry out extraction reaction solution, collection oil reservoir, It uses saturated common salt water washing oil reservoir 3 times again, anhydrous Na₂SO₄Dry oil reservoir, is spin-dried for oil reservoir and crosses column purification, PE/EtOAc elutes white Color intermediate 10；

Step j: the Isosorbide-5-Nitrae dissolved with intermediate 10-dioxane solution is added in reaction under high pressure bottle, is added appropriate 25% Ammonia spirit, autoclave is risen to 98 DEG C, is stirred overnight, is cooled to room temperature, deflates pressure in kettle near 0, takes out reaction Simultaneously low pressure is spin-dried for liquid, and filter cake is collected by filtration in the solid for adding water dispersion to be spin-dried for, and filter cake is repeatedly washed with water, and drying filter cake obtains intermediate Body 11 is not required to purify；

Step k: intermediate 11 (1.0eq), boronic acid derivatives are added into there-necked flaskRaw material (1.1eq), PdCl₂ (dppf) (0.1eq) and K₂CO₃(3.0eq), through N₂Repeatedly after the air of displacement there-necked flask, Isosorbide-5-Nitrae-dioxane/H is added₂O(9/ 1, v/v), be heated to reflux stirring 4h, be filtered to remove insoluble matter with diatomite, collect filtrate, be spin-dried for filtrate, with EtOAc with Water extracts filtrate, collects oil reservoir, then with saturated common salt water washing oil reservoir 3 times, anhydrous Na₂SO₄Dry oil reservoir, is spin-dried for oil reservoir and is used in combination Silicagel column purified pool product Intermediate 12；

Step l: intermediate 12 is added into reaction flask, a small amount of Isosorbide-5-Nitrae-dioxane is added as solvent and enough chlorine Change hydrogen and be saturated Isosorbide-5-Nitrae-dioxane solution, be stirred overnight in 50 DEG C, gradually there is solid generation, TLC monitoring reaction terminates, will be anti- It answers liquid to be cooled to room temperature, filter flask is collected by filtration, and wash filter cake with EtOAc, drain filter cake and collect to obtain product Intermediate 13；

Step m: intermediate 13 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA is slowly added dropwise (5.0eq), solution becomes clarification by muddiness, then is slowly added dropwise(for acyl chlorides or sulfonic acid chloride) (1.5eq), reaction is overnight.Add water Quenching reaction liquid, DCM extraction, saturated salt solution 3 times washings, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain mesh Mark product II.

When raw material isWhen (for acyl chlorides or sulfonic acid chloride), X C, m are 1, using above-mentioned identical Method and steps, obtain general formula III compoundWhen raw material is(for acyl Chlorine or sulfonic acid chloride), X N, m be when being 1, using above-mentioned identical method and steps, obtain VI compound of general formulaWhen raw material isWhen (for acyl chlorides or sulfonic acid chloride), X C, m are 1, use Above-mentioned identical method and steps, obtains compoundWhen raw material is(for acyl chlorides Or sulfonic acid chloride), X C general formulae IV compound obtained using above-mentioned identical method and stepsWork as raw material For(for acyl chlorides or sulfonic acid chloride), X N obtain general formula VIII using above-mentioned identical method and stepsWhen raw material isWhen (for acyl chlorides or sulfonic acid chloride), X C, m are 2, adopt With above-mentioned identical method and steps, general formula VII is obtainedWhen raw material isUsing above-mentioned identical Method and steps obtains compoundThe compound of other similar structure also uses above-mentioned identical method and step Suddenly it is prepared, only changes corresponding raw material.

The present invention also provides above-mentioned 4- amidino-pyridine and azepine ring derivatives include its tautomer, alloisomerism The mixture of body and its all proportions further includes the compound 4- amidino-pyridine that its isotope replaces and azepine ring derivatives.

The present invention also provides above-mentioned 4- amidino-pyridine and azepine ring derivatives pharmaceutically acceptable salt.Wherein with Acid refers at salt, is obtained by the free alkali of parent compound with inorganic acid or reacting for organic acid.Inorganic acid includes hydrochloric acid, hydrogen Bromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid and perchloric acid etc..Organic acid include acetic acid, propionic acid, acrylic acid, oxalic acid, (D) or (L) malic acid, fumaric acid, maleic acid, hydroxybenzoic acid, gamma-hydroxybutyric acid, methoxy benzoic acid, phthalic acid, Methanesulfonic acid, ethanesulfonic acid, naphthalene -1- sulfonic acid, naphthalene-2-sulfonic acid, p-methyl benzenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, almond Acid, succinic acid or malonic acid etc..

The present invention also provides above-mentioned 4- amidino-pyridine and the pharmaceutically acceptable hydrates of azepine ring derivatives.Term " hydrate " indicates further through the compound of active force combination stoichiometry or non-stoichiometric water between Non-covalent molecular.

Pharmaceutical composition is with 4- amidino-pyridine shown in formula Ⅹ, I-VIII and azepine ring derivatives, its salt or its hydration Object is active constituent, addition pharmaceutically acceptable carrier composition.

The present invention also provides above-mentioned 4- amidino-pyridine and azepine ring derivatives and its salt or hydrate, pharmaceutical composition Preparing the purposes in BTK (Bruton ' s tyrosine kinase) inhibitor.

The present invention also provides above-mentioned 4- amidino-pyridine and azepine ring derivatives and its salt or hydrate, pharmaceutical composition Preparing the purposes in BTK (Bruton ' s tyrosine kinase) covalency inhibitor.

The present invention also provides above-mentioned 4- amidino-pyridine and azepine ring derivatives and its salt or hydrate, pharmaceutical composition Purposes in the preparation of antitumor drugs.Described tumour such as breast cancer, colon cancer, lung cancer, lymthoma etc..

The present invention also provides above-mentioned 4- amidino-pyridine and azepine ring derivatives and its salt or hydrate, pharmaceutical composition Preparing the purposes in drugs for rheumatoid arthritis.

The present invention also provides above-mentioned 4- amidino-pyridine and azepine ring derivatives and its salt or hydrate, pharmaceutical composition Purposes in preparation treatment asthmatic medicament.

The present invention also provides above-mentioned 4- amidino-pyridine and azepine ring derivatives and its salt or hydrate, pharmaceutical composition Purposes in preparation treatment Chronic Obstructive Pulmonary Disease drug.

Simultaneously azepine ring derivatives can be used as Bruton ' s tyrosine kinase to 4- amidino-pyridine provided by the invention (BTK) selective covalency inhibitor is applied to a variety of medical usages, such as rheumatoid arthritis, asthma, Chronic Obstructive Pulmonary Disease With the diseases such as tumour, there is good effect.

Detailed description of the invention

Half inhibiting rate IC of Fig. 1 compound III-1 for heart hERG potassium-channel₅₀Value

The clinical arthritis symptom of Fig. 2 III-1 improvement CIA mouse model: the clinical joint of mouse after (A) every group for the treatment of Inflammation marking；(B) after every group for the treatment of before and after mouse pawl photo.The scoring of model group gradually rises at any time it can be seen from Fig. 2A Height, and after drug is treated according to Shandong for Buddhist nun's group and III-1, pathological score gradually decreases.Fig. 2 B can be seen that and normal mouse It compares, model group mouse foot pawl is red and swollen, congested, and disappears for Buddhist nun's group with red and swollen after III-1 treatment by drug according to Shandong, mouse foot Pawl substantially returns to normal mouse state.

Specific embodiment

For convenience of the description to example synthetic route and method below, now by embodiment is raw materials used or the abbreviation system of reagent At the following table 1.

Table 1

(((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3-d] is phonetic by 3- by embodiment 1N- Pyridine -7- base) methyl) phenyl) and acrylamide preparation

Step 1

Raw material 1 (1.0eq) is added into reaction flask, enough THF are added as solvent, are stirred in room temperature, in nitrogen guarantor Raw material NIS (1.1eq) is added portionwise under shield, reaction solution is become to clarify dark brown liquid by suspension after reacting 1h at room temperature. The reaction was continued about 5h or so, is spin-dried for reaction solution, adds water to be dispersed solid and dissolve certain water-solubility impurities, filter is collected by filtration Cake, a large amount of water washing filter cake, drying filter cake obtain product 2.

Step 2

Raw material 2 (1.0eq) is added into reaction flask, nitrobenzyl bromine (1.0eq) and K₂CO₃(1.5eq) is added suitable DMF is stirred in room temperature as solvent, and solution gradually becomes suspension by clarification.TLC monitoring reaction terminates, and adds water quenching reaction, A large amount of water washing filter cake is filtered and used, filter cake is collected and dries to obtain product 3.

Step 3

Raw material 3, suitable Isosorbide-5-Nitrae-dioxane and 25% ammonium hydroxide are added into autoclave, it is anti-in 98 C overnights It answers.Next day and with the cooling reaction kettle of cold water, deflation, taking-up reaction solution and by it be spin-dried for, water-dispersible solid, is collected by filtration solid Body, is repeatedly washed with water filter cake, and drying filter cake obtains product 4.

Step 4

Intermediate 4 (1.0eq), 3,4- methylene-benzene boric acid (1.1eq), PdCl are added into there-necked flask₂(dppf) (0.1eq) and K₂CO₃(3.0eq), through N₂Repeatedly after the air of displacement there-necked flask, it is molten that reaction has been played into there-necked flask with injection Agent 1,4- dioxane/H₂O(9/1,v/v).It is heated to reflux stirring 4h, is filtered to remove insoluble matter with diatomite, collects filtrate, rotation Steaming is spin-dried for filtrate, extracts filtrate with EtOAc and water, collects oil reservoir, then with saturated common salt water washing oil reservoir 3 times, anhydrous Na₂SO₄It is dry Dry oil reservoir is spin-dried for oil reservoir and with silicagel column purified pool product 5.

Step 5

Raw material 5 (1.0eq) is added into reaction flask, NH₄COOH (3.0eq), appropriate MeOH make solvent, add under N2 protection Enter 10%Pd/C (10% mass of raw material 5), be heated to reflux 2h, TLC monitoring reaction terminates.Diatomite is filtered to remove insoluble matter, Filtrate is collected, filtrate is spin-dried for, filtrate is extracted with EtOAc and water, collects oil reservoir, then with saturated common salt water washing oil reservoir 3 times, Anhydrous Na₂SO₄Dry oil reservoir, is spin-dried for oil reservoir up to product 6.

Step 6

Raw material 6 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA (5.0eq) slowly is added dropwise, Solution becomes clarification by muddiness, then acryloyl chloride (1.5eq) slowly is added dropwise, and reaction is overnight.Water quenching is added to go out reaction solution, DCM extraction is satisfied It is washed with saline solution 3 times, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product I-1.¹H NMR (400MHz, DMSO) δ 10.13 (s, 1H), 8.19 (d, J=3.8Hz, 1H), 7.64 (d, J=9.1Hz, 1H), 7.51 (s, 1H), 7.37 (d, J=4.8Hz, 1H), 7.28 (t, J=7.9Hz, 1H), 7.04-6.99 (m, 3H), 6.90 (dd, J=7.9,1.7Hz, 1H), 6.40 (dd, J=17.0,10.1Hz, 3H), 6.23 (dd, J=17.0,2.1Hz, 1H), 6.06 (s, 2H), 5.73 (dd, J =10.1,2.1Hz, 1H), 5.36 (s, 2H) .MS (ESI), m/z:414.26 [M+H]⁺.

Embodiment 2 (S) -1-3- ((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3- D] pyrimidin-7-yl) methyl) piperidin-1-yl) -2- propylene -1- ketone preparation

Embodiment 3 (R) -1-3- ((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3- D] pyrimidin-7-yl) methyl) piperidin-1-yl) -2- propylene -1- ketone preparation

Step 1

Raw material 7 (1.0eq) is added into reaction flask, enough DCM are added as solvent, stirs, is being added in ice bath Et₃It is slowly added dropwise after N (1.5eq) methylsufonyl chloride (1.2eq), solution is changed from clarification shape to suspension, continues to stir 3h TLC monitoring reaction (sulphur is aobvious) afterwards, fully reacting.It is filtered to remove insoluble matter, collects filtrate, repeatedly uses saturated salt solution by extracting Filtrate is washed, the dry oil reservoir of anhydrous sodium sulfate is spin-dried for oil reservoir and obtains oil product 8, without purifying.

Step 2

Raw material 8 (1.0eq) is added into reaction flask, enough ACE are added as solvent, stirs, is slowly added in room temperature LiBr (3.0eq), is gradually heated to reflux state, overnight.Reaction solution is cooled to room temperature, water quenching is added to go out reaction solution, is spin-dried for anti- It is extracted after answering liquid with EtOAc and water, collects oil reservoir, then with saturated common salt water washing oil reservoir 3 times, anhydrous Na₂SO₄Dry oil Layer is spin-dried for oil reservoir up to oil product 9, without purifying.

Step 3

Raw material 2 (1.0eq) is added into reaction flask, 9 (1.0eq) and K₂CO₃Suitable DMF is added as molten in (1.5eq) Agent is stirred in 60 DEG C.TLC monitoring reaction terminates, and adds water and EtOAc to carry out extraction reaction solution, collects oil reservoir, then use saturated common salt Water washing oil reservoir 3 times, anhydrous Na₂SO₄Dry oil reservoir, is spin-dried for oil reservoir and crosses column purification, PE/EtOAc elutes to obtain white intermediate 10.

Step 4

Isosorbide-5-Nitrae dissolved with raw material 10-dioxane solution is added in reaction under high pressure bottle, the ammonium hydroxide of addition appropriate 25% is molten Autoclave is risen to 98 DEG C, is stirred overnight by liquid.It is cooled to room temperature, deflates pressure in kettle near 0, take out reaction solution and low pressure It is spin-dried for, filter cake is collected by filtration in the solid for adding water dispersion to be spin-dried for, and filter cake is repeatedly washed with water, and drying filter cake obtains product 11, is not required to pure Change.

Step 5

Intermediate 11 (1.0eq), 3,4- methylene-benzene boric acid (1.1eq), PdCl are added into there-necked flask₂(dppf) (0.1eq) and K₂CO₃(3.0eq), through N₂Repeatedly after the air of displacement there-necked flask, it is molten that reaction has been played into there-necked flask with injection Agent 1,4- dioxane/H₂O(9/1,v/v).It is heated to reflux stirring 4h, is filtered to remove insoluble matter with diatomite, collects filtrate, rotation Steaming is spin-dried for filtrate, extracts filtrate with EtOAc and water, collects oil reservoir, then with saturated common salt water washing oil reservoir 3 times, anhydrous Na₂SO₄It is dry Dry oil reservoir is spin-dried for oil reservoir and with silicagel column purified pool product 12.

Step 6

Raw material 12 is added into reaction flask, a small amount of Isosorbide-5-Nitrae-dioxane is added as solvent and enough hydrogen chloride saturations Isosorbide-5-Nitrae-dioxane solution, is stirred overnight in 50 DEG C, gradually there is solid generation.TLC monitoring reaction terminates, and reaction solution is cooling To room temperature, filter flask is collected by filtration, and washs filter cake with EtOAc, drains filter cake and collects to obtain product 13.

Step 7

Raw material R-13 or S-13 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA is slowly added dropwise (5.0eq), solution become clarification by muddiness, then acryloyl chloride (1.5eq) slowly is added dropwise, and reaction is overnight.Water quenching is added to go out reaction solution, DCM extraction, saturated salt solution 3 times washings, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product R-II- 1 and S-II-1.

R-II-1:¹H NMR (400MHz, DMSO) δ 8.12 (s, 1H), 7.29 (s, 1H), 7.00 (dd, J=8.4, 4.8Hz, 2H), 6.90 (dd, J=7.9,1.7Hz, 1H), 6.74 (dd, J=22.2,9.0Hz, 1H), 6.04 (d, J= 17.3Hz, 5H), 5.63 (d, J=11.8Hz, 1H), 4.11 (m, 1H), 4.07 (d, J=7.6Hz, 2H), 3.90-3.77 (m, 1H), 3.10 (t, J=11.2Hz, 1H), 2.67 (t, J=11.2Hz, 1H), 2.03 (d, J=11.8Hz, 1H), 1.68 (m, 2H), 1.26 (d, J=9.7Hz, 2H) .MS (ESI), m/z:406.22 [M+H]⁺.

S-II-1:¹H NMR (400MHz, DMSO) δ 8.12 (d, J=5.5Hz, 1H), 7.29 (s, 1H), 7.01 (dt, J= 4.7,3.1Hz, 2H), 6.91 (dd, J=7.9,1.7Hz, 1H), 6.75 (ddd, J=12.2,9.6,6.8Hz, 1H), 6.25- 5.91 (m, 5H), 5.62 (t, J=11.6Hz, 1H), 4.13 (dd, J=8.3,5.0Hz, 1H), 4.06 (d, J=7.6Hz, 2H), 3.84 (dd, J=25.1,12.9Hz, 1H), 3.08 (t, J=11.5Hz, 1H), 2.67 (t, J=11.5Hz, 1H), 2.13- 2.00(m,1H),1.68(s,2H),1.32–1.22(m,2H).MS(ESI),m/z:406.23[M+H]⁺

Embodiment 4 (S) -3- (3- ((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3- D] pyrimidin-7-yl) methyl) piperidin-1-yl) propionitrile preparation (S-II-2)

Embodiment 5 (R) -3- (3- ((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3- D] pyrimidin-7-yl) methyl) piperidin-1-yl) propionitrile preparation (R-II-2)

Raw material R-13 or S-13 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA is slowly added dropwise (5.0eq), solution become clarification by muddiness, then 3- bromopropionitrile (1.5eq) slowly is added dropwise, and reaction is overnight.Water quenching is added to go out reaction solution, DCM extraction, saturated salt solution 3 times washings, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product R-II- 2 and S-II-2.

R-II-2:¹H NMR (400MHz, DMSO) δ 8.12 (s, 1H), 7.26 (s, 1H), 7.01 (d, J=7.9Hz, 2H), 6.91 (dd, J=9.8,2.0Hz, 1H), 6.04 (d, J=17.5Hz, 4H), 4.11-4.01 (m, 2H), 2.67 (m, 1H), 2.60 (t, J=7.0Hz, 2H), 2.13 (m, 1H), 2.05 (t, J=9.6Hz, 1H), 1.94 (t, J=9.6Hz, 1H), 1.64 (s, 1H), 1.47 (s, 1H), 1.38 (d, J=12.6Hz, 1H), 1.24 (m, 2H), 1.01 (d, J=9.4Hz, 1H), 0.84 (d, J= 8.3Hz,1H).MS(ESI),m/z:405.22[M+H]⁺.

S-II-2:¹H NMR (400MHz, DMSO) δ 8.12 (d, J=5.6Hz, 1H), 7.26 (s, 1H), 7.03-6.98 (m, 2H), 6.91 (dd, J=8.1,1.6Hz, 1H), 6.80-6.69 (m, 1H), 6.18-6.00 (m, 4H), 4.08-4.04 (m, 2H), 2.66 (dd, J=10.3,3.9Hz, 1H), 2.61 (d, J=6.9Hz, 2H), 2.12 (d, J=3.2Hz, 1H), 2.09- 2.01 (m, 1H), 1.98-1.91 (m, 1H), 1.65 (dd, J=9.1,3.6Hz, 1H), 1.52-1.45 (m, 1H), 1.41-1.33 (m, 1H), 1.27-1.21 (m, 2H), 1.05-0.96 (m, 1H), 0.84 (d, J=7.7Hz, 1H) .MS (ESI), m/z:405.21 [M+H]⁺.

Embodiment 6 (S) -3- (3- ((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3- D] pyrimidin-7-yl) methyl) piperidin-1-yl) acetonitrile preparation (S-II-3)

Embodiment 7 (R) -3- (3- ((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3- D] pyrimidin-7-yl) methyl) piperidin-1-yl) acetonitrile preparation (R-II-3)

Raw material R-13 or S-13 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA is slowly added dropwise (5.0eq), solution become clarification by muddiness, then chloroacetonitrile (1.5eq) slowly is added dropwise, and reaction is overnight.Water quenching is added to go out reaction solution, DCM Extraction, saturated salt solution 3 times washings, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elute target product R-II-3 and S-II-3。

R-II-3:¹H NMR(400MHz,DMSO)δ8.13(s,1H),7.27(s,1H),7.04–6.99(m,2H),6.91 (dd, J=7.8,1.8Hz, 1H), 6.06 (s, 4H), 4.04 (t, J=6.1Hz, 2H), 3.67 (s, 2H), 2.76 (d, J= 11.1Hz, 2H), 2.08 (dd, J=11.3,9.6Hz, 2H), 1.86 (ddd, J=11.2,7.5,3.7Hz, 1H), 1.53 (d, J =11.2Hz, 2H), 1.32-1.20 (m, 2H) .MS (ESI), m/z:391.20 [M+H]⁺.

S-II-3:¹H NMR (400MHz, DMSO) δ 8.15 (s, 1H), 7.30 (s, 1H), 7.00 (dd, J=6.8, 4.8Hz, 2H), 6.92-6.89 (m, 1H), 6.23 (s, 2H), 6.06 (s, 2H), 4.08 (d, J=7.2Hz, 2H), 3.67 (s, 2H), 2.66 (dd, J=8.7,6.7Hz, 1H), 2.58-2.52 (m, 1H), 2.17 (t, J=9.6Hz, 2H), 2.07-1.99 (m, 1H), 1.73-1.66 (m, 1H), 1.56 (d, J=13.5Hz, 1H), 1.01 (dd, J=20.2,10.5Hz, 1H), 0.86-0.80 (m,1H).MS(ESI),m/z:391.23[M+H]⁺.

Embodiment 8 (S) -1-3- ((4- amino -5- (4- phenyl) -7H- pyrroles [2,3-d] pyrimidin-7-yl) methyl) Piperidin-1-yl) -2- propylene -1- ketone (B3-15) preparation (S-II-4)

S-II-4:1H NMR (400MHz, DMSO) δ 8.15 (s, 1H), 7.47 (d, J=8.5Hz, 2H), 7.41 (t, J= 7.8Hz, 2H), 7.34 (s, 1H), 7.16 (t, J=7.4Hz, 1H), 7.10 (t, J=7.5Hz, 4H), 6.81-6.57 (m, 1H), 6.14 (s, 2H), 6.04 (d, J=16.6Hz, 1H), 5.63 (d, J=10.7Hz, 1H), 4.19-4.05 (m, 3H), 3.92- 3.77 (m, 1H), 3.10 (s, 1H), 2.67 (s, 1H), 2.07 (d, J=35.0Hz, 1H), 1.69 (s, 2H), 1.28 (s, 2H)

Embodiment 9 (R) -1-3- ((4- amino -5- (4- phenyl) -7H- pyrroles [2,3-d] pyrimidin-7-yl) methyl) Piperidin-1-yl) -2- propylene -1- ketone preparation (R-II-4)

R-II-4:¹H NMR (400MHz, DMSO) δ 8.15 (s, 1H), 7.47 (d, J=8.5Hz, 2H), 7.41 (t, J= 7.9Hz, 2H), 7.34 (s, 1H), 7.16 (t, J=7.4Hz, 1H), 7.10 (t, J=7.5Hz, 4H), 6.70 (ddd, J= 60.4,16.3,10.6Hz, 1H), 6.14 (s, 2H), 6.04 (d, J=16.7Hz, 1H), 5.62 (t, J=11.1Hz, 1H), 4.21-4.05 (m, 3H), 3.92-3.77 (m, 1H), 3.15-2.85 (m, 1H), 2.67 (t, J=11.3Hz, 1H), 2.07 (d, J =34.2Hz, 1H), 1.69 (s, 2H), 1.26 (d, J=18.4Hz, 2H)

10 1- of embodiment (4- ((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3-d] Pyrimidin-7-yl) methyl) piperidin-1-yl) and -2- propylene -1- ketone preparation (III-1)

Step 1

Raw material 14 (1.0eq) is added into reaction flask, enough DCM are added as solvent, stirs, is being added in ice bath Et₃It is slowly added dropwise after N (1.5eq) methylsufonyl chloride (1.2eq), solution is changed from clarification shape to suspension, continues to stir 3h TLC monitoring reaction (sulphur is aobvious) afterwards, fully reacting.It is filtered to remove insoluble matter, collects filtrate, repeatedly uses saturated salt solution by extracting Filtrate is washed, the dry oil reservoir of anhydrous sodium sulfate is spin-dried for oil reservoir and obtains oil product 15, without purifying.

Step 2

Raw material 15 (1.0eq) is added into reaction flask, enough ACE are added as solvent, stirs, is slowly added in room temperature LiBr (3.0eq), is gradually heated to reflux state, overnight.Reaction solution is cooled to room temperature, water quenching is added to go out reaction solution, is spin-dried for anti- It is extracted after answering liquid with EtOAc and water, collects oil reservoir, then with saturated common salt water washing oil reservoir 3 times, anhydrous Na₂SO₄Dry oil Layer is spin-dried for oil reservoir up to oil product 16, without purifying.

Step 3

Raw material 2 (1.0eq) is added into reaction flask, 16 (1.0eq) and K₂CO₃Suitable DMF is added as molten in (1.5eq) Agent is stirred in 60 DEG C.TLC monitoring reaction terminates, and adds water and EtOAc to carry out extraction reaction solution, collects oil reservoir, then use saturated common salt Water washing oil reservoir 3 times, anhydrous Na₂SO₄Dry oil reservoir, is spin-dried for oil reservoir and crosses column purification, PE/EtOAc elutes to obtain white intermediate 17.

Step 4

Isosorbide-5-Nitrae dissolved with raw material 17-dioxane solution is added in reaction under high pressure bottle, the ammonium hydroxide of addition appropriate 25% is molten Autoclave is risen to 98 DEG C, is stirred overnight by liquid.It is cooled to room temperature, deflates pressure in kettle near 0, take out reaction solution and low pressure It is spin-dried for, filter cake is collected by filtration in the solid for adding water dispersion to be spin-dried for, and filter cake is repeatedly washed with water, and drying filter cake obtains product 18, is not required to pure Change.

Step 5

Intermediate 18 (1.0eq), 3,4- methylene-benzene boric acid (1.1eq), PdCl are added into there-necked flask₂(dppf) (0.1eq) and K₂CO₃(3.0eq), through N₂Repeatedly after the air of displacement there-necked flask, it is molten that reaction has been played into there-necked flask with injection Agent 1,4- dioxane/H₂O(9/1,v/v).It is heated to reflux stirring 4h, is filtered to remove insoluble matter with diatomite, collects filtrate, rotation Steaming is spin-dried for filtrate, extracts filtrate with EtOAc and water, collects oil reservoir, then with saturated common salt water washing oil reservoir 3 times, anhydrous Na₂SO₄It is dry Dry oil reservoir is spin-dried for oil reservoir and with silicagel column purified pool product 19.

Step 6

Raw material 19 is added into reaction flask, a small amount of Isosorbide-5-Nitrae-dioxane is added as solvent and enough hydrogen chloride saturations Isosorbide-5-Nitrae-dioxane solution, is stirred overnight in 50 DEG C, gradually there is solid generation.TLC monitoring reaction terminates, and reaction solution is cooling To room temperature, filter flask is collected by filtration, and washs filter cake with EtOAc, drains filter cake and collects to obtain product 20.

Step 7

Raw material 20 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA (5.0eq) slowly is added dropwise, Solution becomes clarification by muddiness, then acryloyl chloride (1.5eq) slowly is added dropwise, and reaction is overnight.Water quenching is added to go out reaction solution, DCM extraction is satisfied It is washed with saline solution 3 times, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product III-1.

¹H NMR (400MHz, DMSO) δ 8.12 (s, 1H), 7.26 (s, 1H), 7.00 (dd, J=6.9,4.8Hz, 2H), 6.90 (dd, J=7.9,1.7Hz, 1H), 6.77 (dd, J=16.7,10.5Hz, 1H), 6.25-5.95 (m, 5H), 5.65 (d, J =2.4Hz, 1H), 4.38 (d, J=12.7Hz, 1H), 4.05 (d, J=7.2Hz, 2H), 4.01 (s, 1H), 2.98 (s, 1H), 2.57 (d, J=12.9Hz, 1H), 2.16 (dd, J=7.4,3.5Hz, 1H), 1.54 (d, J=12.2Hz, 2H), 1.14 (d, J= 23.7Hz,2H).MS(ESI),m/z:406.22[M+H]⁺.

11 3- of embodiment (4- ((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3-d] Pyrimidin-7-yl) methyl) piperidin-1-yl) and propionitrile preparation (III-2)

Raw material 20 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA (5.0eq) slowly is added dropwise, Solution becomes clarification by muddiness, then 3- bromopropionitrile (1.5eq) slowly is added dropwise, and reaction is overnight.Water quenching is added to go out reaction solution, DCM extraction is satisfied It is washed with saline solution 3 times, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product III-2.¹H NMR (400MHz, DMSO) δ 8.12 (s, 1H), 7.25 (s, 1H), 7.00 (dd, J=6.5,4.8Hz, 2H), 6.90 (dd, J=7.9, 1.7Hz, 1H), 6.06 (s, 4H), 4.03 (d, J=7.2Hz, 2H), 2.83 (d, J=11.4Hz, 2H), 2.62 (t, J= 6.5Hz,2H),2.53(s,1H),1.94–1.78(m,3H),1.52–1.43(m,2H),1.30–1.18(m,2H),0.88– 0.79(m,1H).MS(ESI),m/z:406.24[M+H]⁺.

12 2- of embodiment (4- ((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3-d] Pyrimidin-7-yl) methyl) piperidin-1-yl) and acetonitrile preparation (III-3)

Raw material 20 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA (5.0eq) slowly is added dropwise, Solution becomes clarification by muddiness, then chloroacetonitrile (1.5eq) slowly is added dropwise, and reaction is overnight.Water quenching is added to go out reaction solution, DCM extraction, saturation Saline solution 3 times washings, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product III-3.¹H NMR (400MHz, DMSO) δ 8.13 (s, 1H), 7.27 (s, 1H), 7.04-6.99 (m, 2H), 6.91 (dd, J=7.8,1.8Hz, 1H), 6.06 (s, 4H), 4.04 (t, J=6.1Hz, 2H), 3.67 (s, 2H), 2.76 (d, J=11.1Hz, 2H), 2.08 (dd, J= 11.3,9.6Hz, 2H), 1.86 (ddd, J=11.2,7.5,3.7Hz, 1H), 1.53 (d, J=11.2Hz, 2H), 1.32-1.20 (m,2H).MS(ESI),m/z:391.20[M+H]⁺.

Embodiment 13-34

The synthetic operation of other derivatives is same as above, and only change participates in the boric acid raw material of Suzuki reaction, is as a result shown such as The following table 2:

Table 2

Hydrogen modification embodiment on piperidine ring N:

35 5- of embodiment (benzo [d] [1,3] dioxolanes -5- base) -7- ((1- ethylene sulfuryl) piperidin-4-yl) first Base) -7H- pyrroles [2,3-d] pyrimidine -4- amine preparation (III-26)

Raw material 20 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA (5.0eq) slowly is added dropwise, Solution becomes clarification by muddiness, then 3- chlorine sulfonic acid chloride (1.5eq) slowly is added dropwise, and reaction is overnight.Water quenching is added to go out reaction solution, DCM extraction, Saturated salt solution 3 times washings, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product III-26.¹H NMR (400MHz, DMSO) δ 8.12 (s, 1H), 7.26 (s, 1H), 7.01 (d, J=8.8Hz, 2H), 6.94-6.86 (m, 1H), 6.75 (dd, J=16.5,10.0Hz, 1H), 6.26-6.01 (m, 6H), 4.07 (d, J=7.2Hz, 2H), 3.51 (d, J=12.0Hz, 2H), 2.57 (t, J=11.3Hz, 2H), 1.99 (s, 1H), 1.60 (d, J=12.0Hz, 2H), 1.35-1.21 (m, 2H) .MS (ESI),m/z:442.23[M+H]⁺.

Embodiment 36 (E) -1- (4- ((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2, 3-d] pyrimidin-7-yl) methyl) piperidin-1-yl)-4- dimethylamino) and-2- butene-1 -one preparation (III-27)

(E) -4- (dimethylamino) -2- butenoate hydrochlorate (1.0eq) and appropriate DCM are added into reaction flask as molten Agent sequentially adds DIEA (1.0eq) in ice bath, PyBOP (1.5eq) and DIEA (1.5eq), stirs 5-10min, and it is molten that mixing is added dropwise Liquid 20 (1.0eq)+DIEA (4.0eq)+DCM (appropriate), reaction is stayed overnight at room temperature.DCM extraction, saturated salt solution 3 times washings, nothing Aqueous sodium persulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product III-27.¹H NMR(400MHz,DMSO)δ8.12 (s, 1H), 7.26 (s, 1H), 7.00 (dd, J=7.8,4.7Hz, 2H), 6.90 (dd, J=7.9,1.7Hz, 1H), 6.67 (d, J =15.2Hz, 1H), 6.61-6.51 (m, 1H), 6.04 (s, 4H), 4.38 (d, J=12.5Hz, 1H), 4.05 (dd, J=7.2, 2.8Hz, 2H), 4.01 (d, J=7.1Hz, 1H), 3.25 (d, J=5.9Hz, 2H), 2.99 (t, J=11.7Hz, 1H), 2.58 (t, J=11.7Hz, 1H), 2.31 (s, 6H), 2.16 (d, J=12.2Hz, 1H), 1.55 (d, J=11.4Hz, 2H), 1.12 (d, J=6.0Hz, 2H) .MS (ESI), m/z:463.14 [M+H]⁺.

37 5- of embodiment (4- phenyl) -7- ((1- ethylene sulfuryl) piperidin-4-yl) methyl) -7H- pyrroles [2,3-d] The preparation (III-28) of pyrimidine -4- amine

Raw material 21 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA (5.0eq) slowly is added dropwise, Solution becomes clarification by muddiness, then 3- chlorine sulfonic acid chloride (1.5eq) slowly is added dropwise, and reaction is overnight.Water quenching is added to go out reaction solution, DCM extraction, Saturated salt solution 3 times washings, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product III-28.¹H NMR (400MHz, DMSO) δ 8.15 (s, 1H), 7.51-7.38 (m, 4H), 7.32 (s, 1H), 7.16 (t, J=7.4Hz, 1H), 7.10 (dd, J=8.1,6.0Hz, 4H), 6.75 (dd, J=16.5,10.0Hz, 1H), 6.09 (dd, J=18.3,13.3Hz, 2H), 4.09 (d, J=7.1Hz, 2H), 3.52 (d, J=12.4Hz, 2H), 2.57 (t, J=11.3Hz, 2H), 2.00 (d, J= 9.7Hz, 1H), 1.60 (d, J=11.5Hz, 2H), 1.28 (dd, J=25.2,9.4Hz, 2H)

Embodiment 38 (E) -1- (4- ((4- amino -5- (4- oxygroup) phenyl) -7H- pyrroles [2,3-d] pyrimidin-7-yl) first Base) piperidin-1-yl) -4- dimethylamino) and -2- propylene -1- ketone preparation (III-29)

(E) -4- (dimethylamino) -2- butenoate hydrochlorate (1.0eq) and appropriate DCM are added into reaction flask as molten Agent sequentially adds DIEA (1.0eq) in ice bath, PyBOP (1.5eq) and DIEA (1.5eq), stirs 5-10min, and it is molten that mixing is added dropwise Liquid 21 (1.0eq)+DIEA (4.0eq)+DCM (appropriate), reaction is stayed overnight at room temperature.DCM extraction, saturated salt solution 3 times washings, nothing Aqueous sodium persulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product III-29.¹H NMR(400MHz,DMSO)δ8.14 (s, 1H), 7.50-7.39 (m, 5H), 7.31 (s, 1H), 7.16 (t, J=7.4Hz, 1H), 7.13-7.07 (m, 4H), 6.58 (d, J=6.3Hz, 1H), 6.12 (s, 2H), 4.38 (d, J=11.6Hz, 1H), 4.07 (d, J=7.2Hz, 2H), 4.01 (d, J= 12.7Hz, 1H), 3.09 (d, J=5.0Hz, 2H), 2.98 (t, J=12.4Hz, 1H), 2.57 (t, J=12.4Hz, 1H), 2.20 (s, 6H), 1.55 (d, J=12.0Hz, 2H), 1.18-1.05 (m, 2H) .MS (ESI), m/z:511.41 [M+H]⁺.

Embodiment 39-48

Specific synthetic method such as compound III series, compound IV structure and nuclear magnetic data table 3 presented below:

Table 3

((4- amino -5- (benzo [d] [1,3] dioxolanes -5- base) -7H- pyrroles [2,3-d] is phonetic by 2- by embodiment 49N- Pyridine -7- base) ethyl) acrylamide preparation

Step 1

Raw material 36 (1.0eq) is added into reaction flask and suitable super dry DMF makees solvent, DIEA (5.0eq) slowly is added dropwise, Solution becomes clarification by muddiness, then acryloyl chloride (1.5eq) slowly is added dropwise.TLC monitoring reaction terminates, and adds water quenching to go out reaction solution, DCM Extraction, saturated salt solution 3 times washings, anhydrous sodium sulfate is dry, is spin-dried for column, MeOH/DCM elutes to obtain target product V-1.¹H NMR (400MHz, DMSO) δ 8.26 (t, J=5.4Hz, 1H), 8.14 (s, 1H), 7.22 (s, 1H), 7.01 (d, J=7.9Hz, 1H), 6.96 (d, J=1.5Hz, 1H), 6.88 (dd, J=7.9,1.6Hz, 1H), 6.17 (dd, J=17.1,9.9Hz, 2H), 6.12-5.99 (m, 4H), 5.58 (dd, J=9.9,2.4Hz, 1H), 4.25 (t, J=6.1Hz, 2H), 3.55 (dd, J=12.0, 6.0Hz,2H).MS(ESI),m/z:352.13[M+H]⁺.

Embodiment 50-53

Specific synthetic method such as compound II series, compound VI structure and nuclear magnetic data table 4 presented below:

Table 4

Embodiment 54-57

Specific synthesis such as compound II series, compound VII structure and nuclear magnetic data table 5 presented below:

Table 5

Pharmacodynamics test part

The present invention also provides the experiment of the Pharmacological Activity Screening of above-mentioned part of compounds, i.e., external biochemistry level inhibits BTK to swash Enzymatic activity experiment.

Material: BTK kinases (Carna)；Polypeptide FAM-P22 (GL Biochem)；ATP, DMSO and EDTA (Sigma)；96 Orifice plate (Corning), positive control staurosporine (Sigma).

Method:

1, the alkali buffer and stop buffer of 1x kinases are equipped with

1) the 1x kinases alkali buffer of BTK includes 50Mm hydroxyethyl piperazine second thiosulfonic acid (HEPES), Ph 7.5； 0.0015%Brij-35；10Mm MgCl₂；2Mm DTT.

2) stop buffer includes 100 mMs of HEPES, PH 7.5；0.015%Brij-35,50Mm EDTA.

2, the preparation of compound

1) test-compound is configured to 50 times of highest test concentrations with 100% DMSO.It is dilute to shift 100 μ l compounds It releases in liquid to orifice plate.

2) in 100%DMSO to 2 holes for increasing by 100 μ l, and the plate is denoted as original plate.10 μ l are shifted from original plate Compound is to one piece of 96 new orifice plate and is denoted as intermediate plate, and the 1x kinases alkali buffering of 90 μ l is added into each hole of intermediate plate Liquid, and plank, which is placed on shaking table, is uniformly mixed compound solution with 1x kinases alkali buffer.5 μ l are respectively taken from intermediate plate again Multiple holes are formed on mixture to 96 orifice plates, and the plate is denoted as detection plate.

3, enzyme reaction

1) 2.5x enzyme solutions are equipped with, kinases is added in the kinases alkali buffer of 1x.

2) it is equipped with 2.5x polypeptide buffer, FAM- tag polypeptide and ATP are added in the kinases alkali buffer of 1x.

3) transfer 2.5x enzyme solutions are into detection plate.Compound of each hole of detection plate equipped with 5 μ l comprising 10%DMSO Solution adds the 2.5x enzyme solutions of 10 μ l, in 25 DEG C of incubation 10min.

4) 2.5x polypeptide solution is added into each hole of detection plate, the end of 25 μ l is added after being incubated for suitable duration in 28 DEG C Only buffer is to terminate enzyme reaction.

4, the initial data in every hole is read and recorded, and initial data is converted accordingly.

1) inhibiting rate=(maximum value-compound conversion value)/(maximum value-minimum value) * 100, wherein maximum value DMSO Control group data, minimum value are the blank value that enzyme is not added.

2) calculation of half inhibitory concentration IC₅₀Value, with log [administration concentration] for abscissa, inhibiting rate is ordinate, A dose-effect curve is fitted in Graphpad Prism 5, obtains drug concentration when its 50% inhibiting rate, i.e., thus IC of the compound on Kinase levels₅₀Value.

The following table 6 provides average IC of the compounds of this invention about BTK₅₀Range, wherein " A " indicates IC₅₀Value is less than 10nM, " B " indicate IC₅₀For value between 10nM and 100nM, " C " indicates IC₅₀Between 100nM and 1000nM, " D " is indicated value IC₅₀Value is greater than 1000nM.

Table 6

Embodiment	BTK IC₅₀/nM	Embodiment	BTK IC₅₀/nM
				Staurosporine	141	III-20	C
I-1	C	III-21	C
				R-II-1	C	III-22	C
R-II-2	D	III-23	C
				R-II-3	D	III-24	B
R-II-4	A	III-25	B
				S-II-1	C	III-26	A
S-II-2	D	III-27	D
				S-II-3	D	III-28	A
S-II-4	A	III-29	B
				III-1	B	IV-1	A
III-2	D	IV-2	A
				III-3	D	IV-3	B
III-4	A	VI-4	A
				III-5	B	IV-5	A
III-6	C	IV-6	A
				III-7	C	IV-7	D
III-8	D	IV-8	D
				III-9	B	IV-9	D
III-10	C	V-1	B
				III-11	D	VI-1	B
III-12	D	VI-2	A
				III-13	C	VI-3	A
III-14	D	VI-4	A
				III-15	C	VII-1	C
III-16	D	VII-2	C
				III-17	C	VII-3	C
III-18	B	VII-4	B
				III-19	C

Correlation data analysis, many compounds inhibit the bioactivity of BTK preferable, such as R-II-4, III-1, III-5, III-26, III-28, IV-1, IV-2, IV-4, IV-5, IV-6, VI-2, VI-3, VI-4 etc..Meanwhile we are to these compounds The measurement of water-soluble has been carried out at room temperature, when what is connected on skeleton pyrrole ring or pyrazole ring 3 is pepper cyclic group, The dissolubility of compound is substantially improved compared with Biphenyl Ether substituent, the former solubility is about 0.1mg/ml, and the latter is then less than 0.01mg/ml provides good basic condition for the oral administration of later period animal model.

We assess the activity of part of compounds in a variety of non-small cell lung cancer wild types and mutant cell system. Meanwhile it being accredited as effective BTK inhibitor and will also investigate their anti-proliferative capacities to B cell lymphoma cell line.

Lines: NSCLC wild-type cell system A549 and mutant cell system H1975, HCC827 is equal From biological therapy National Key Laboratory, Sichuan University cell bank.Cell culture is in containing 10% fetal calf serum, penicillin, chain In the RPMI1640 culture medium of mycin, in 5%CO₂Incubator in 37 DEG C of constant temperature incubations.Cell is collected after counting, is pressed 3000-4000/hole is plated in 96 orifice plates, for attached cell, after cell is adherent, by the way that various concentration is added The compound of (10000,2000,400,80,8,0.8nM) handles 72h, and control wells add isometric culture medium.Processing time point arrives Afterwards, MTT is added and is incubated for 1-3h, after the blue precipitate that observation MTT effect generates under the microscope is obvious, discard the culture containing MTT Base is added DMSO by the amount of every 150 μ l of hole and sufficiently dissolves precipitating, in OD value 570nm wavelength detecting light absorption value.By looking for formula: suppression Rate processed=1- control wells light absorption value/experimental port light absorption value, calculates each experimental group inhibiting rate.Compound concentration for the treatment of is converted into again Logarithmic form, corresponding inhibiting rate 6.0 software fitted dose of Graphpad prism-response curve, and it is fitted IC50 Value.

B cell lymphoma cell line: B cell lymphoma cell line Jeko-1, SUDHL-4, SUDHL-6, OCI-LY1, HBL-1, Daudi, raji, MV4-11 and Ramos are both from biological therapy National Key Laboratory, Sichuan University cell bank.Carefully After born of the same parents' culture is adherent, the compound that gradient concentration compound (50,10,2,0.4,0.08,0.016 μM) are added handles 72h, control Hole adds isometric culture medium, and post-processing is same as above.

MCF-7, HEPG2, A549, H1975, HCC827 derive from Sichuan University's biological therapy National Key Laboratory cell Library, cell culture is in the DMEM culture medium of penicillin and streptomysin containing 10% fetal calf serum and 100U/ml.

Selected compounds are wild to EGFR/the anti-proliferative capacity IC of the NSCLC cell line of mutant₅₀(μM) see the table below 7.

Table 7

Above the results showed that most compounds are active to EGFR wild type and drug resistant tumour cell.It is selected Anti-proliferative capacity IC of the compound to B cell lymphoma cell line₅₀(Μ m) see the table below 8.

Table 8

Anti-proliferative capacity IC of the selected compounds to B cell lymphoma cell line₅₀(μM) see the table below 9.

Table 9

Anti-proliferative capacity IC of the selected compounds to other tumor cell lines₅₀(μM) see the table below 10.

Table 10

From table 9 and table 10 it can also be seen that above-mentioned multiple compounds show there is work to B cell lymphoma and solid tumor Property.

Since RA is a kind of non-life-threatening disease, still cytotoxicity test has been carried out to part of compounds. HEK293, LO2 and THP-1 cell are provided by four big university student's object treatment National Key Laboratories, as a result see the table below 11.

Table 11

Compound

HEK293

LO2

THP-1

Compound

HEK293

LO2

THP-1

Buddhist nun is replaced according to Shandong

19.3±1.2

17.4±0.5

20.1±0.7

III-28

2.7±0.5

3.2±0.4

3.3±0.4

R-II-4

2.9±0.2

11.7±1.9

8.8±1.0

III-29

4.8±0.8

6.6±0.9

3.2±1.1

S-II-4

1.3±0.2

14.2±2.1

10.7±1.5

VI-2

2.8±0.3

10.1±2.1

14.4±0.9

III-1

34.2±1.9

31.3±2.1

38.9±1.5

VI-3

11.2±1.1

10.9±1.6

15.7±2.2

III-4

13.3±0.9

16.0±1.1

16.7±1.0

VI-4

1.6±0.2

19.1±3.5

18.3±2.7

III-26

2.5±0.4

3.2±0.4

3.7±0.7

VII-3

10.2±1.6

15.2±2.0

18.9±2.8

Therefrom, we pick compound III-1 and have carried out cardiac toxic test, in CHO cell line (the middle bright Kant of traditional Chinese medicines Company's cell seed bank) on measurement to the inhibiting effect of hERG potassium current.The cell that this experiment uses is stable transfection hERG The Chinese hamster ovary celI of cDNA, the cell express the channel P25 of hERG.Cell culture is in containing 10% fetal calf serum, 100 μ g/mL hygromycin In the Ham's F12 culture medium of B and 100 μ g/mL Geneticins, incubator environment is 37 DEG C and 5%CO₂.When cell is in logarithm Growth period and cell density are tested when reaching 80%-90%.Before the experiments, cell is gently blown and beaten at room temperature After 15-20 times, after being handled 3-5 minutes at 37 DEG C with Detachin (cell dissociation reagent, a kind of trypsase substituting agent), Cell is resuspended in the serum free medium containing HEPES (25mM) buffer and CHO-S-SFM II.Compound III-1 sun Property control Cisapride be dissolved in DMSO, the concentration for experiment is respectively III-1 (0.4,1.2,3.7,11.1,33.3 He 100mM), Cisapride (3,1,0.3,0.1,0.03 and 0.01mM), is added separately to the Chinese hamster ovary celI of stable transfection hERG cDNA In, after 37 DEG C are incubated for 72 hours, its fluorescent value is measured, the data of cell use automatic QPatch (Sophion at room temperature Biosciences, Denmark) it is collected.The data of acquisition use measurement software (the measurement software V5.0 provided by Sophion Version), Microsoft Excel and Graphpad Prism 5.0 is analyzed.The result shows that: IC of the III-1 to hERG₅₀For 11.10 μM, much higher than the IC for replacing Buddhist nun according to Shandong₅₀=0.97 μM, show that III-1 almost without toxic effect, will not cause heart Heart cross quiver, heart rate is too fast etc..

Meanwhile we utilize and determine compound on SD rat (6-8 weeks, 200g or so, purchased from magnificent Fukang animal company) The bioavilability of III-1.By the way that rat is divided into two groups, every group 5,3mg/kg III-1 and mouth are given by tail vein After clothes give 3mg/kgIII-1 compound, it is intubated by arteria carotis, after administration 5min, 15min, 30min, 1h, 2h, 4h, 6h, 8h, 10h, 12h, and take 400 microlitres of blood respectively for 24 hours, and blood sample takes upper serum, pass through by being centrifuged after adding heparin sodium LC-MS-MS detects drug in blood serum concentration, the curve after vein and oral administration are calculated by software software DAS 2.0 Lower peak area (AUC) obtains the oral administration biaavailability of III-1 compound, and the compound bioavailability is up to 49.15%, mouth Taking half-life period is 7.04h, is superior to marketed drug according to Shandong for Buddhist nun, can be administered orally, the results are shown in Table 12.

12 III-1 of table is in SD rat pharmacokinetic parameters

	Intravenous injection	It is oral
			It measures (mg/kg)	3	3
C_max(μg/L)	3132.28	257.82
			t_max(h)	0.11	0.96
t_1/2(h)	5.18	7.04
			AUC_0-t(μg/L*h)	2522.28	1239.63
V,V/F(L/kg)	1.21(V)	26.15(V/F)
			Cl,Cl/F(L/h/kg)	8.23V(Cl)	2.75(Cl/F)
F%		49.15

The properties such as enzymatic activity, bioavilability and solubility according to compound III-1, with reference to being replaced according to Shandong for report For Buddhist nun to the therapeutic scheme of arthritic mice, we establish the therapeutic effect that arthritis model investigates compound III-1.It takes 10 weeks Age male DBA/1J mouse, weight about 18-20g are purchased from Beijing HFK Bio-Technology Co., Ltd..First by two type glue of ox Former CII is dissolved in the acetic acid of 0.05M by 2mg/ml, 4 DEG C, is protected from light, and is stirred overnight sufficiently to dissolve, then by its with it is isometric Complete Freund's adjuvant (CFA) containing 1mg/ml BCG vaccine mixes in equal volume.With tee tube connect two 20ml BD syringes into Row emulsification, until emulsion droplet is dropped on clear water and do not scatter.Using 10% chloraldurate by DBA/1j mouse anesthesia, by 0.15ml/ It is intradermal that the ox Type Ⅱ collagen and CFA that have emulsified completely are only injected in mouse tail root, point 3 points of injections, the immune same day is denoted as the 1 day.The 21st day after initial immunity, by by the ox Type Ⅱ collagen and incomplete Freund's adjuvant IFA of the emulsification of identical method, press It is intradermal after second of booster immunization that the CII emulsified completely and IFA are injected in mouse tail root by 0.75ml/ mouse, The changes of weight of mouse is monitored, three-times-weekly；Meanwhile whether the joint by monitoring mouse is red and swollen or stiff, analyzes rheumatoid The development progress and severity of property arhritis conditions, and mouse joint is faced according to double-blind study according to red and swollen stiffness Bed scoring^[51].Each claw score is between 0-4, therefore every mouse top score is 16 points.Obvious appearance is selected by scoring The mouse of arthritic.Mouse is divided into four groups, every group of 8 mouse, including negative control group (unmodeled group), model group, Positive drug, for Buddhist nun's group (20mg/kg is administered orally daily) III-1 treatment group (20mg/kg is administered orally daily), is examined according to Shandong Examine the therapeutic effect of drug.By treatment in 30 days by a definite date, the arthritic mice foot swelling of Liang Ge treatment group was eased even It disappears, the therapeutic effect of compound III-1 is suitable with positive drug.

Claims

1.4- amidino-pyridine and azepine ring derivatives, structural formula is as shown in formula Ⅹ:

Wherein, X is C or N；

R₂ForSubstituted or unsubstituted nitrogenous 5 membered heterocycloalkyl orR₃ForOr replace Or unsubstituted nitrogenous 6 membered heterocycloalkyl；Substituent group in nitrogenous 5 membered heterocycloalkyl, nitrogenous 6 membered heterocycloalkyl ring is independent It is-H, C1~C8 alkyl, halogen or-CN；R₄, substituent group, nitrogenous 6 membered heterocycloalkyl nitrogen on nitrogenous 5 membered heterocycloalkyl nitrogen On substituent group be independentlyR₅、R₆Be independently C2~C8 alkenyl, C2~C8 alkynyl,R₇、R₈It is independently-H or C1~C8 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

R₁ForThe cycloalkenyl of C3~C8,Or substituted or unsubstituted 5~ 10 unit's heteroaryls；The hetero atom of 5~10 unit's heteroaryl is that hetero atom is N, O, S, and hetero atom number is 1~3；Described 5 The substituent group of~10 unit's heteroaryls is-H, halogen or C1~C8 alkyl；R₉~R₁₃It is independently-H, halogen, C1~C8 alkyl, C1 ~C8 alkoxy ,-CF₃、-OCF₃、Phenyl, phenoxy group, benzyloxy, C1~C8 oxygen carbonyl,C is 0~4；Or R₁₀And R₁₁Combination forms 3~8 circle heterocyclic ring alkane Base or 3~8 membered unsaturated heterocycles；3~8 membered heterocycloalkyl, 3~8 membered unsaturated heterocycles hetero atom be N, O, S, miscellaneous original Sub- number is 1~3.

2. 4- amidino-pyridine according to claim 1 and azepine ring derivatives, it is characterised in that: X is C or N；

R₂ForSubstituted or unsubstituted nitrogenous 5 membered heterocycloalkyl orR₃ForOr replace Or unsubstituted nitrogenous 6 membered heterocycloalkyl；Substituent group in nitrogenous 5 membered heterocycloalkyl, nitrogenous 6 membered heterocycloalkyl ring is independent It is-H, C1~C6 alkyl, halogen or-CN；R₄, substituent group, nitrogenous 6 membered heterocycloalkyl nitrogen on nitrogenous 5 membered heterocycloalkyl nitrogen On substituent group be independentlyR₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl,R₇、R₈It is independently-H or C1~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

R₁ForThe cycloalkenyl of C3~C6,Or substituted or unsubstituted 5~ 8 unit's heteroaryls；The hetero atom of 5~8 unit's heteroaryl is that hetero atom is N, O, S, and hetero atom number is 1~3；Described 5~8 The substituent group of unit's heteroaryl is-H, halogen or C1~C6 alkyl；R₉~R₁₃It is independently-H, halogen, C1~C6 alkyl, C1~C6 Alkoxy ,-CF₃、-OCF₃、Phenyl, phenoxy group, benzyloxy, C1~C6 oxygen carbonyl,C is 0~4；Or R₁₀And R₁₁Combination forms 3~6 circle heterocyclic rings Alkyl or 3~6 membered unsaturated heterocycles；3~6 membered heterocycloalkyl, 3~6 membered unsaturated heterocycles hetero atom be N, O, S, it is miscellaneous Atom number is 1~3；

Preferably, X is C or N；R₂ForOrR₃ForR₁₅~R₁₇、R₁₉~R₂₁、R₂₃~ R₂₆、R₂₈~R₃₁、R₃₃~R₃₆It is independently-H, C1~C6 alkyl, halogen or-CN；R₄、R₁₄、R₁₈、R₂₂、R₂₇、R₃₂It is independentlyR₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl, R₇、R₈It is independently-H or C1~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

It is further preferred that X is C or N；R₂ForR₃ForR₁₅~R₁₇、R₁₉~R₂₁、R₂₃~ R₂₆、R₂₈~R₃₁、R₃₃~R₃₆It is independently-H, C1~C4 alkyl, halogen or-CN；R₄、R₁₄、R₁₈、R₂₂、R₂₇、R₃₂It is independentlyR₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~3；N is 0~3；Z, p is 0~3；

R₁ForThe cycloalkenyl of C3~C6,Or substituted or unsubstituted 5~ 6 unit's heteroaryls；The hetero atom of 5~6 unit's heteroaryl is that hetero atom is N, O, S, and hetero atom number is 1~2；Described 5~6 The substituent group of unit's heteroaryl is-H, halogen or C1~C4 alkyl；R₉~R₁₃It is independently-H, halogen, C1~C4 alkyl, C1~C4 Alkoxy ,-CF₃、-OCF₃、Phenyl, phenoxy group, benzyloxy, C1~C4 oxygen carbonyl,C is 0~3；Or R₁₀And R₁₁Combination forms 3~6 circle heterocyclic rings Alkyl or 3~6 membered unsaturated heterocycles；3~6 membered heterocycloalkyl, 3~6 membered unsaturated heterocycles hetero atom be N, O, S, it is miscellaneous Atom number is 1~2；

Still further preferably, X is C or N；R₂ForR₃ ForR₁₅~R₁₇、R₁₉~R₂₁、R₂₃ ~R₂₆、R₂₈~R₃₁、R₃₃~R₃₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₄、R₁₄、R₁₈、R₂₂、R₂₇、R₃₂ It is independentlyR₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Still more preferably, X is C or N；R₂ForR₃ ForR₁₅~R₁₇、R₁₉~R₂₁、R₂₃~ R₂₆、R₂₈~R₃₁、R₃₃~R₃₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₄、R₁₄、R₁₈、R₂₂、R₂₇、R₃₂Solely It is vertical to beR₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

R₁ForThe cycloalkenyl of C3~C6,Or substituted or unsubstituted 6 Membered nitrogen-containing heteroaryl base；The substituent group of the 6 membered nitrogen-containing heteroaryl base is-H, halogen or C1~C4 alkyl；R₉~R₁₃Be independently- H ,-F ,-Cl ,-Br, C1~C4 alkyl, C1~C4 alkoxy ,-CF₃、-OCF₃、Phenyl, phenoxy group, benzyl Oxygroup, C1~C4 oxygen carbonyl,C is 0~2；Or R₁₀And R₁₁ Combination forms 3~6 membered heterocycloalkyls or 3~6 membered unsaturated heterocycles；The hetero atom of 3~6 membered heterocycloalkyl is O, hetero atom Number is 1~2；The hetero atom of 3~6 membered unsaturated heterocycle is N, and hetero atom number is 1~2；

Most preferably, R₂ForR₃For R₁₅~R₁₇、R₁₉~R₂₁、R₂₃~R₂₆、R₂₈~R₃₁、R₃₃~ R₃₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₄、R₁₄、R₁₈、R₂₂、R₂₇、R₃₂It is independentlyR₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl, R₇、R₈It is independently-H or C1~C4 alkyl；M is 1 or 2；N is 1 or 2；Z is 0；P is 1 or 2；

3. 4- amidino-pyridine according to claim 1 and azepine ring derivatives, it is characterised in that: work as R₂Form It is 1, R₃ForWhen X is C, structural formula is as shown in formula I:

R₄ForR₅、R₆Be independently C2~C8 alkenyl, C2~C8 alkynyl, R₇、R₈It is independently-H or C1~C8 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

4. 4- amidino-pyridine according to claim 3 and azepine ring derivatives, it is characterised in that: R₄For R₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl,R₇、R₈It is independently-H or C1~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

Preferably, R₄ForR₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~3；N is 0~3；Z, p is 0~3；

Still further preferably, R₄ForR₅、R₆It is independently C2~C4 alkenyl, C2~C4 Alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Still more preferably, R₄ForR₅、R₆It is independently C2~C4 alkenyl, C2~C4 Alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Most preferably, R₄ForR₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1 or 2；N is 1 or 2；Z is 0；P is 1 Or 2；

5. 4- amidino-pyridine according to claim 2 and azepine ring derivatives, it is characterised in that: work as R₂Form It is 1, R₃ForWhen X is C, structural formula is as shown in formula II:

Wherein, R₂₈~R₃₁It is independently-H, C1~C6 alkyl, halogen or-CN；R₂₇For R₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl,R₇、R₈It is independently-H or C1 ~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

R₁ForThe cycloalkenyl of C3~C6,Or substituted or unsubstituted 5~ 8 unit's heteroaryls；The hetero atom of 5~8 unit's heteroaryl is that hetero atom is N, O, S, and hetero atom number is 1~3；Described 5~8 The substituent group of unit's heteroaryl is-H, halogen or C1~C6 alkyl；R₉~R₁₃It is independently-H, halogen, C1~C6 alkyl, C1~C6 Alkoxy ,-CF₃、-OCF₃、Phenyl, phenoxy group, benzyloxy, C1~C6 oxygen carbonyl,C is 0~4；Or R₁₀And R₁₁Combination forms 3~6 circle heterocyclic rings Alkyl or 3~6 membered unsaturated heterocycles；3~6 membered heterocycloalkyl, 3~6 membered unsaturated heterocycles hetero atom be N, O, S, it is miscellaneous Atom number is 1~3.

6. 4- amidino-pyridine according to claim 5 and azepine ring derivatives, it is characterised in that: R₂₈~R₃₁Be independently- H, C1~C4 alkyl, halogen or-CN；R₂₇ForR₅、R₆It is independently C2~C4 alkenyl, C2 ~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~3；N is 0~3； Z, p is 0~3；

Preferably, R₂₈~R₃₁It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₇For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independent It is-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Still more preferably, R₂₈~R₃₁It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₇For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、 R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Most preferably, R₂₈~R₃₁It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₇For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independent It is-H or C1~C4 alkyl；M is 1 or 2；N is 1 or 2；Z is 0；P is 1 or 2；

7. 4- amidino-pyridine according to claim 2 and azepine ring derivatives, it is characterised in that: work as R₂Form It is 1, R₃ForWhen X is C, structural formula is as shown in formula III:

Wherein, R₂₃~R₂₆It is independently-H, C1~C6 alkyl, halogen or-CN；R₂₂For R₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl,R₇、R₈It is independently-H or C1 ~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

8. 4- amidino-pyridine according to claim 7 and azepine ring derivatives, it is characterised in that: R₂₃~R₂₆Be independently- H, C1~C4 alkyl, halogen or-CN；R₂₂ForR₅、R₆It is independently C2~C4 alkenyl, C2 ~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~3；N is 0~3； Z, p is 0~3；

Preferably, R₂₃~R₂₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₂For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independent It is-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

R₁ForThe cycloalkenyl of C3~C6,Or substituted or unsubstituted 5~6 Unit's heteroaryl；The hetero atom of 5~6 unit's heteroaryl is that hetero atom is N, O, S, and hetero atom number is 1~2；Described 5~6 The substituent group of unit's heteroaryl is-H, halogen or C1~C4 alkyl；R₉~R₁₃It is independently-H, halogen, C1~C4 alkyl, C1~C4 Alkoxy ,-CF₃、-OCF₃、Phenyl, phenoxy group, benzyloxy, C1~C4 oxygen carbonyl,C is 0~3；Or R₁₀And R₁₁Combination forms 3~6 circle heterocyclic rings Alkyl or 3~6 membered unsaturated heterocycles；3~6 membered heterocycloalkyl, 3~6 membered unsaturated heterocycles hetero atom be N, O, S, it is miscellaneous Atom number is 1~2；

Still more preferably, R₂₃~R₂₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₂For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、 R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Most preferably, R₂₃~R₂₆It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₂₂For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independent It is-H or C1~C4 alkyl；M is 1 or 2；N is 1 or 2；Z is 0；P is 1 or 2；

9. 4- amidino-pyridine according to claim 2 and azepine ring derivatives, it is characterised in that: work as R₂Form It is 2, R₃ForWhen X is C, structural formula is as shown in formula VII:

10. 4- amidino-pyridine according to claim 9 and azepine ring derivatives, it is characterised in that: R₂₃~R₂₆It is independent For-H, C1~C4 alkyl, halogen or-CN；R₂₂ForR₅、R₆It is independently C2~C4 alkene Base, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~3；N is 0 ~3；Z, p is 0~3；

11. 4- amidino-pyridine according to claim 2 and azepine ring derivatives, it is characterised in that: work as R₂ForWhen X is C, structural formula is as shown in formula IV:

Wherein, R₁₅~R₁₇It is independently-H, C1~C6 alkyl, halogen or-CN；R₁₄For R₅、R₆Be independently C2~C6 alkenyl, C2~C6 alkynyl,R₇、R₈It is independently-H or C1 ~C6 alkyl；M is 1~4；N is 0~4；Z, p is 0~4；

12. 4- amidino-pyridine according to claim 11 and azepine ring derivatives, it is characterised in that: R₁₅~R₁₇It is independent For-H, C1~C4 alkyl, halogen or-CN；R₁₄ForR₅、R₆It is independently C2~C4 alkene Base, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~3；N is 0 ~3；Z, p is 0~3；

Preferably, R₁₅~R₁₇It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₁₄For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、R₈It is independent It is-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Still more preferably, R₁₅~R₁₇It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₁₄For R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、 R₈It is independently-H or C1~C4 alkyl；M is 1~2；N is 0~2；Z, p is 0~2；

Most preferably, R₁₅~R₁₇It is independently-H, C1~C4 alkyl ,-F ,-Cl ,-Br or-CN；R₁₄It is independently R₅、R₆Be independently C2~C4 alkenyl, C2~C4 alkynyl,R₇、 R₈It is independently-H or C1~C4 alkyl；M is 1 or 2；N is 1 or 2；Z is 0；P is 1 or 2；

13. 4- amidino-pyridine according to claim 2 and azepine ring derivatives, it is characterised in that: work as R₂ForWhen X is N, structural formula is as shown in formula VIII:

14. 4- amidino-pyridine according to claim 13 and azepine ring derivatives, it is characterised in that: R₁₅~R₁₇It is independent For-H, C1~C4 alkyl, halogen or-CN；R₁₄ForR₅、R₆It is independently C2~C4 alkene Base, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~3；N is 0 ~3；Z, p is 0~3；

15. 4- amidino-pyridine according to claim 2 and azepine ring derivatives, it is characterised in that: work as R₂Form It is 1, R₃ForWhen X is N, structural formula is as shown in formula VI:

16. 4- amidino-pyridine according to claim 15 and azepine ring derivatives, it is characterised in that: R₂₃~R₂₆It is independent For-H, C1~C4 alkyl, halogen or-CN；R₂₂ForR₅、R₆It is independently C2~C4 alkene Base, C2~C4 alkynyl,R₇、R₈It is independently-H or C1~C4 alkyl；M is 1~3；N is 0 ~3；Z, p is 0~3；

17. described in any item 4- amidino-pyridines and azepine ring derivatives, structural formula are as follows according to claim 1~16:

18. the described in any item 4- amidino-pyridines of claim 1~17 and azepine ring derivatives pharmaceutically acceptable salt.

19. the described in any item 4- amidino-pyridines of claim 1~17 and the pharmaceutically acceptable hydration of azepine ring derivatives Object.

20. pharmaceutical composition is wanted by the described in any item 4- amidino-pyridines of claim 1~17 and azepine ring derivatives, right Hydrate described in salt described in asking 18 or claim 19 is active constituent, addition pharmaceutically acceptable carrier composition.

21. salt described in the described in any item 4- amidino-pyridines of claim 1~17 and azepine ring derivatives, claim 18, Pharmaceutical composition described in hydrate described in claim 19 or claim 20 is preparing the purposes in BTK inhibitor.

22. salt described in the described in any item 4- amidino-pyridines of claim 1~17 and azepine ring derivatives, claim 18, Pharmaceutical composition described in hydrate described in claim 19 or claim 20 is preparing the use in BTK covalency inhibitor On the way.

23. salt described in the described in any item 4- amidino-pyridines of claim 1~17 and azepine ring derivatives, claim 18, Pharmaceutical composition described in hydrate described in claim 19 or claim 20 treats tumour, rheumatoid arthrosis in preparation Purposes in the drug of the diseases such as inflammation, asthma, Chronic Obstructive Pulmonary Disease.