CN109983015A - 6- pyrazoles-[1,2,4] triazol [4,3-a] pyridine -3- amide derivatives, preparation method and its application in medicine - Google Patents

Abstract

6- pyrazoles-[1 shown in logical formula (I), 2,4] triazol [4,3-a] pyridine -3- amide derivatives, preparation method and pharmaceutical composition containing the derivative and its as therapeutic agent, purposes especially as TGF-β receptor kinase inhibitor and preparation treatment, prevention or reduce the tumour mediated by TGF-β overexpression drug in purposes.

Description

6- pyrazoles-[1,2,4] triazol [4,3-a] pyridine -3- amide derivatives, preparation method and its application in medicine Technical field

The invention belongs to field of medicaments, it is related to a kind of new 6- pyrazoles-[1,2,4] triazol [4,3-a] pyridine -3- amide derivatives, preparation method and pharmaceutical composition containing the derivative and its as therapeutic agent especially as the purposes of the inhibitor of TGF-β receptor kinase and preparation treatment, prevention or reduce the tumour mediated by TGF-β overexpression drug in purposes.

Background technique

Transforming growth factor TGF-β (Transforming Growth Factor β) is a member in dimeric polypeptide growth factor superfamily, it includes such as activin, inhibin, Bone Morphogenetic Protein (Bone morphogenetic proteins, BMPs), Growth and Differentiation Factors (Growth differentiation factors,) and Miao's Le Shi pipe mortifier (M ü llerian-inhibiting substance, MIS) GDFs.

TGF-β has 3 three kinds of TGF-β 1, TGF-β 2 and TGF-β hypotypes, they are participated in, and cell Proliferation and differentiation, wound healing, extracellular matrix generates and immunosuppressive regulation.Referring to such as Massague, J.Ann.Rev, Cell.Biol.6:594-641 (1990)；Roberts,A.B.Peptide Growth Factor and Their receptors,95:419-472Berlin:Springer-Verlag(1990)；Roberts, A.B. and Sporn M.B.Growth Factor 8:1-9 (1993)；And Alexandrow, M.G., Moses, H.L.Cancer Res.55:1452-1457 (1995).Three kinds of hypotypes of TGF-β are present in most of cell together with its receptor.Every kind of TGF-β hypotype is synthesized as precursor protein, which is cracked into C- end regions (latency-associatedpeptide latency associated peptide, LAP) and N- end part in the cell, referred to as mature or active TGF-β.Before secreting in cell, LAP is generally connect with mature T GF- β in non-covalent mode.LAP-TGF- β complex cannot be in conjunction with TGF-β receptor and without biological activity.It includes that (and active) is released from complex for example, with thrombospondin-1 or fibrinolytic enzyme interacting that TGF-β, which usually passes through number of mechanisms,.TGF-β 1 passes through two kinds high conservative single membrane span serine/threonine kinase transduction signal, i.e. I type (ALK5) and II type TGF-β receptor.Once ligand induced oligomerization, the serine/threonine residue in the area II receptor Hyperphosphorylationof ALK5GS, the binding site by creating Smad albumen cause the activation of ALK5.The ALK5 of the activation then Smad2 and Smad3 albumen at the SSXS- motif of the end phosphorylation C-, dissociates from receptor, and generate Heterogeneous Composite body (heteromeric complex) with Smad4 so as to cause them.The transposition of Smad complex is assembled, the transcription of final activating cell epimatrix component and matrix-degrading proteases enzyme inhibitor in core with specific DNA-binding co-factor and auxiliary adjusting control agent.

The reason of hyperactive of TGF-β signal path is many human diseases (excess deposition of such as extracellular matrix, the unusual high levels of inflammatory response, fibrosis conditions and progressive carcinoma).Advanced stage various tumours, stroma cell generally over-expresses TGF-β in tumour cell and tumour.This causes the stimulation of angiogenesis and cell movement, the inhibition of immune system and the interaction of tumour cell and extracellular matrix to increase (such as Hojo, M. et al., Nature 397:530-534 (1999)).So tumour cell becomes more have invasion, distant organs are transferred to for example, Maehara, Y. et al., J.Clin.Oncol.17:607-614 (1999)；Picon, A. et al., Cancer Epidemiol.Biomarkers Prev.7:497-504 (1998)).

The glomerulus that many experiments zooscopy demonstrates TGF-β is expressed and being associated between fibrosis, 5/6 rat remnant model of Thy-1 rat model, rabbit-anti-GBM glomerulonephritis and focal segmental glomerulosclerosis including proliferative glomerulonephritis, have recently commentary (such as, Bitzer, M. etc., Kidney Blood Press.Res.21:1-12 (1998)).E.g., the neutralizing antibody of TGF-β improves the glomerulus histology (Border, W.A. et al., Nature 346:371-374 (1990)) in Thy-1 Nephritis Model.

TGF-β 1 and its receptor are overexpressed in damaged blood vessels and fibroproliferative vascular damage, cause the excessive generation of extracellular matrix (for example, Saltis, J. et al., Clin.Exp.Pharmacol.Physiol.23:193-200 (1996)；McCaffrey, T.A. et al., J.Clin.Invest.96:2667-2675 (1995)).

TGF-β 2 is horizontal have in most of aqueous humor tumour eyes with juvenile glaucoma and in almost half eye with primary open angle glaucoma (POAG) increase (such as, Picht, G. et al., Graefes Arch.Clin.Exp.Ophthalmol.239:199-207 (2001)).

Therefore, it is desirable to develop the inhibitor to TGF-β family member to prevent and/or treat the disease including this signal path.Regulator (such as antagonist) patent application of disclosed TGF-β family member receptor includes WO2004111046, WO2012000595, WO2012002680, WO2013009140, WO2016106266.

In order to achieve the purpose that better therapeutic effect, better meet the market demand, the present invention will provide a kind of TGF-β receptor kinase inhibitor of new structural high-efficiency low-toxicity, and find to may make the compound of this class formation to have the characteristics that medicine for well-behaved by introducing amide groups.

Summary of the invention

The purpose of the present invention is to provide a kind of logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt,

Or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt,

Wherein:

Ring A is aryl or heteroaryl；

R ¹Selected from hydrogen atom, alkyl, halogenated alkyl, hydroxyl, hydroxyalkyl, amino, naphthenic base, heterocycle, aryl, heteroaryl ,-C (O) OR ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸, wherein alkyl, naphthenic base, heterocycle, aryl and the heteroaryl are optionally selected from halogen, alkyl, alkoxy, halogenated alkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, naphthenic base, heterocycle, aryl, heteroaryl ,-C (O) OR each independently ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸In one or more substituent groups replaced；

R ²It is identical or different, and it is each independently selected from hydrogen atom, halogen, alkyl, alkoxy, halogenated alkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, naphthenic base, heterocycle, aryl, heteroaryl ,-C (O) OR ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸；

R ³It is identical or different, and it is each independently selected from hydrogen atom, halogen, alkyl, alkoxy, halogenated alkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, naphthenic base, heterocycle, aryl and heteroaryl；

R ⁴And R ⁵It is each independently selected from hydrogen atom, alkyl, halogenated alkyl, naphthenic base, heterocycle, aryl and heteroaryl, wherein alkyl, naphthenic base, heterocycle, aryl and the heteroaryl are each independently optionally replaced one or more substituent groups in alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, naphthenic base, heterocycle, aryl and heteroaryl；

Alternatively, the R ⁴And R ⁵Heterocycle is formed together with the nitrogen-atoms being connected, wherein in the heterocycle containing 1~2 identical or different hetero atom selected from N, O and S, and the heterocycle is optionally replaced one or more substituent groups in alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, naphthenic base, heterocycle, aryl and heteroaryl；

R ⁶Selected from hydrogen atom, alkyl, amino, halogenated alkyl, naphthenic base, heterocycle, aryl and heteroaryl；

R ⁷And R ⁸It is each independently selected from hydrogen atom, alkyl, halogenated alkyl, naphthenic base, heterocycle, aryl and heteroaryl, wherein alkyl, naphthenic base, heterocycle, aryl and the heteroaryl are each independently optionally replaced one or more substituent groups in alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, naphthenic base, heterocycle, aryl and heteroaryl；

Alternatively, the R ⁷And R ⁸Heterocycle is formed together with the nitrogen-atoms being connected, wherein in the heterocycle containing 1~2 identical or different hetero atom selected from N, O and S, and the heterocycle is optionally replaced one or more substituent groups in alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, naphthenic base, heterocycle, aryl and heteroaryl；

N is 0,1,2,3 or 4；

S is 0,1,2 or 3；And

M is 0,1 or 2.

In a preferred embodiment in accordance with this invention, the logical formula (I) compound represented, middle ring A are heteroaryl, preferably 5- or 6-membered heteroaryl, more preferable pyridyl group.

In a preferred embodiment in accordance with this invention, the logical formula (I) compound represented, to lead to formula (II) compound represented:

Or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt,

Wherein: R ¹~R ⁵, n and s be as defined in logical formula (I).

In a preferred embodiment in accordance with this invention, the logical formula (I) compound represented, wherein R ⁴And R ⁵It is hydrogen atom.

In a preferred embodiment in accordance with this invention, the logical formula (I) compound represented, to lead to formula (III) compound represented:

Or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt,

Wherein: R ¹And R ²As defined in logical formula (I).

In a preferred embodiment in accordance with this invention, the logical formula (I) compound represented, wherein R ¹Selected from hydrogen atom, alkyl, naphthenic base and heterocycle, preferably hydrogen atom, C _1-6Alkyl, 3 to 6 yuan of naphthenic base, 3 to 6 circle heterocyclic ring bases, more preferable hydrogen atom, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, tetrahydrofuran base or THP trtrahydropyranyl.

In a preferred embodiment in accordance with this invention, the logical formula (I) compound represented, wherein R ²For hydrogen atom or alkyl, preferably hydrogen atom, methyl, ethyl, propyl, isopropyl or butyl.

In a preferred embodiment in accordance with this invention, the logical formula (I) compound represented, wherein R ³For hydrogen atom.

The compound of the present invention includes its all conformer, such as cis and trans isomer；And its all optical isomer and stereoisomer and its mixture.The compound of the present invention has asymmetric center, therefore there are different enantiomters and diastereoisomers.The present invention relates to the purposes of the compounds of this invention, and can use and contain their all pharmaceutical compositions and treatment method.The present invention relates to the purposes of all such isomers and its mixture.

Typical compound of the invention includes but is not limited to:

Or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt.

Another aspect of the present invention relates to a kind of methods for preparing logical formula (I) compound represented, this method comprises:

The compound reaction of the compound and general formula (I-B) of general formula (I-A), obtains the compound of logical formula (I),

Wherein:

W is boronate or 4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base；

X is halogen, preferably bromine；

Ring A, R ¹~R ⁵, n and s be as defined in logical formula (I).

Another aspect of the present invention relates to a kind of methods for preparing logical formula (I) compound represented, this method comprises:

The compound reaction of the compound and general formula (I-Bb) of general formula (I-Aa), obtains the compound of logical formula (I),

Wherein:

W is boronate or 4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base；

X is halogen, preferably bromine；

Ring A, R ¹~R ⁵, n and s be as defined in logical formula (I).

Another aspect of the present invention relates to a kind of pharmaceutical compositions, described pharmaceutical composition contain treatment effective dose logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or pharmaceutical salt and one or more pharmaceutically acceptable carriers, diluent or excipient.The invention further relates to a kind of methods for preparing above-mentioned composition comprising by logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt mixed with one or more pharmaceutically acceptable carriers, diluent or excipient.

The invention further relates to logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or comprising its pharmaceutical composition, preparing the purposes in the drug for inhibiting TGF-β (especially mankind's TGF-β) signal transduction path.

The invention further relates to logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or comprising its pharmaceutical composition, in preparation for treating, preventing or reducing the purposes in the drug that tumour cell (especially human tumor cell) shifts.

The invention further relates to logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or purposes comprising its pharmaceutical composition in drug of the preparation for treating, preventing or reducing the tumour mediated by TGF-β overexpression, the purposes in the drug of the tumour for over-expressing mediation by TGF-β is treated, prevents or reduced especially by mankind's TGF-β signal transduction path is inhibited.

In text of the invention, the treatment, prevention or the disease for mitigating (the especially mankind) are selected from: cardiovascular disease, all kinds of inflammation, tumour, the fibrosis of the various causes of disease, injury of blood vessel, nephrosis, hepatosis, tuberculosis, adult respiratory distress syndrome (ARDS), intimal thickening, eye disease, arteriopathy or hypertrophica corium scar or keloid during the wound healing caused by wound or wound occurs to be formed, peritonaeum and sub-dermal adhesion, chorionitis, fibrosclerosis, progressive systemic sclerosis, osteoporosis, ulcer, nervous function lowers, male erectile dysfunction, Peyronie's disease, dupuytren's contracture, Alzheimer's disease and Raynaud's syndrome.

The invention further relates to logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or comprising its pharmaceutical composition, purposes in the drug of above-mentioned disease of the preparation for treating, preventing or mitigating (the especially mankind).

The invention further relates to it is a kind for the treatment of, prevention or reduce human tumor cell transfer method comprising give required bacterium logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or the pharmaceutical composition comprising it.

The invention further relates to a kind for the treatment of, prevention or the methods for reducing the tumour mediated by TGF-β overexpression, especially by inhibit TGF-β signal transduction path come treat, prevent or reduces by TGF-β overexpression mediation tumour method comprising give required bacterium logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or the pharmaceutical composition comprising it.

The invention further relates to a kind for the treatment of, prevention or mitigate the method that (the especially mankind) be selected from above-mentioned disease comprising give required bacterium logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or the pharmaceutical composition comprising it.

The invention further relates to it is a kind of inhibit TGF-β (especially mankind's TGF-β) signal transduction path method comprising give required bacterium logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or the pharmaceutical composition comprising it.

The invention further relates to a kind of logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or include its pharmaceutical composition, drugs with function.

The invention further relates to a kind of logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or include its pharmaceutical composition, it acts on TGF-β receptor kinase inhibitor, especially TGF-β receptor I (TGF-β RI) kinase inhibitor.

The invention further relates to a kind of logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or comprising its pharmaceutical composition, be used to treat, prevent or reduce tumour cell (especially human tumor cell) transfer.

The invention further relates to a kind of logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or include its pharmaceutical composition, it is used to treat, prevents or reduce the tumour mediated by TGF-β overexpression, and the tumour mediated by TGF-β overexpression is treated, prevents or reduced especially by TGF-β signal transduction path is inhibited.

The invention further relates to a kind of logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its officinal salt or comprising its pharmaceutical composition, be used to treat, prevent or mitigate the above-mentioned disease of (the especially mankind).

Pharmaceutical composition containing active constituent, which can be, is suitable for oral form, such as tablet, dragee, pastille, water or oil suspension, dispersible powder or particle, lotion, hard or soft capsule or syrup or elixir.Orally administered composition can be prepared according to any known method for preparing Pharmaceutical composition in this field, such composition can contain one or more ingredients selected from the following: sweetener, corrigent, colorant and preservative, to provide pleasing and palatable pharmaceutical formulation.Tablet contains active constituent and the suitable nontoxic pharmaceutical excipient for preparing tablet for mixing.These excipient can be inert excipient, granulating agent, disintegrating agent, adhesive and lubricant,.These tablets can not be coated or can be by the taste for covering drug or delay disintegration and absorption in the gastrointestinal tract, thus the known technology for providing slow releasing function in a long time is coated.

The Perle offer oral preparation that wherein active constituent is mixed with inert solid diluent or in which active constituent with water-solubility carrier or oily solvent is also provided.

Water slurry contains active material and the suitable excipient for preparing water slurry for mixing.Such excipient is suspending agent, dispersing agent or wetting agent.Aqueous suspension can also contain one or more preservatives, one or more colorants, one or more corrigents and one or more sweeteners.

Oil suspension can active constituent be suspended in vegetable oil or mineral oil is formulated by making.Oil suspension can contain thickener.Above-mentioned sweetener and corrigent can be added, to provide palatable preparation.These compositions can be saved by the way that antioxidant is added.

Pharmaceutical composition of the invention is also possible to the form of oil in water emulsion.Oil mutually can be or mixtures thereof vegetable oil or mineral oil.Suitable emulsifier can be naturally-produced phosphatide, and emulsion can also contain sweetener, corrigent, preservative and antioxidant.Such preparation can also contain moderator, preservative, colorant and antioxidant.

Pharmaceutical composition of the invention can be sterile injectable aqueous form.The acceptable solvent or solvent that can be used have water, ringer's solution and isotonic sodium chlorrde solution.Aseptic injection preparation can be wherein active constituent be dissolved in the aseptic injection oil-in-water microemulsion of oily phase can be by a large amount of injections in part, will be in the blood flow of injection or micro emulsion injection patient.Alternatively, preferably giving solution and micro emulsion in the way of it can keep the compounds of this invention constant circulating concentration.To keep this constant density, continuous intravenous delivery device can be used.The example of this device is Deltec CADD-PLUS.TM.5400 type Iv pump.

Pharmaceutical composition of the invention can be for intramuscular and the aseptic injection water of subcutaneous administration or the form of oil suspension.Can be by known technology, the dispersing agent or wetting agent and suspending agent for being suitable for those described above prepare the suspension.Aseptic injection preparation is also possible to the aseptic injectable solution or suspension prepared in the acceptable non-toxic diluent of parenteral or solvent.Furthermore, it is convenient to use sterile fixed oil as solvent or suspension media.For this purpose, any reconciliation fixing oil can be used.In addition, fatty acid can also prepare injection.

The compounds of this invention can be given by the suppository form for rectally.Can by by drug be at normal temperatures solid but be in the rectum liquid, thus can dissolve in the rectum and discharge the suitable nonirritant excipient mixing of drug to prepare these pharmaceutical compositions.

As known to those skilled in the art, the dosage of drug depend on many factors, including but be not limited to following factor: the activity of particular compound used, the age of patient, the weight of patient, the health status of patient, the behavior of patient, the diet of patient, administration time, administration mode, the rate of excretion, combination of drug etc.；In addition, optimal therapeutic modality can be verified such as mode, the consumption per day of general formula compound (I) or the type of pharmaceutical salt for the treatment of according to traditional therapeutic scheme.

Detailed description of the invention

Unless stated to the contrary, the term used in the specification and in the claims has following meanings.

Term " alkyl " refers to saturated aliphatic hydrocarbons group, is the linear chain or branched chain group comprising 1 to 20 carbon atom, preferably comprises the alkyl of 1 to 12 carbon atom, the alkyl of further preferably 1 to 6 carbon atom.Non-limiting example includes methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group, tert-butyl, sec-butyl, n-pentyl, 1, 1- dimethyl propyl, 1, 2- dimethyl propyl, 2, 2- dimethyl propyl, 1- ethyl propyl, 2- methyl butyl, 3- methyl butyl, n-hexyl, 1- Ethyl-2-Methyl propyl, 1, 1, 2- thmethylpropyl, 1, 1- dimethylbutyl, 1, 2- dimethylbutyl, 2, 2- dimethylbutyl, 1, 3- dimethylbutyl, 2- ethyl-butyl, 2- methyl amyl, 3- methyl amyl, 4- methyl amyl, 2, 3- dimethylbutyl, n-heptyl, 2- methylhexyl, 3- methylhexyl, 4- methylhexyl, 5- methylhexyl, 2, 3- dimethyl amyl group, 2, 4- dimethyl amyl group, 2, 2- bis- Methyl amyl, 3, 3- dimethyl amyl group, 2- ethylpentyl, 3- ethylpentyl, n-octyl, 2, 3- dimethylhexanyl, 2, 4- dimethylhexanyl, 2, 5- dimethylhexanyl, 2, 2- dimethylhexanyl, 3, 3- dimethylhexanyl, 4, 4- dimethylhexanyl, 2- ethylhexyl, 3- ethylhexyl, 4- ethylhexyl, 2- methyl -2- ethylpentyl, 2- methyl -3- ethylpentyl, n-nonyl, 2- methyl -2- ethylhexyl, 2- methyl -3- ethylhexyl, 2, 2- diethyl amyl group, positive decyl, 3, 3- diethylhexyl, 2, 2- diethylhexyl, and its various branched isomers etc..Low alkyl group more preferably containing 1 to 6 carbon atom, non-limiting embodiment includes methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group, tert-butyl, sec-butyl, n-pentyl, 1, 1- dimethyl propyl, 1, 2- dimethyl propyl, 2, 2- dimethyl propyl, 1- ethyl propyl, 2- methyl butyl, 3- methyl butyl, n-hexyl, 1- Ethyl-2-Methyl propyl, 1, 1, 2- thmethylpropyl, 1, 1- dimethylbutyl, 1, 2- dimethylbutyl, 2, 2- dimethylbutyl, 1, 3- dimethylbutyl, 2- ethyl-butyl, 2- methyl amyl, 3- methyl amyl, 4- methyl amyl, 2, 3- dimethylbutyl etc..Alkyl can be substituted or non-substituted, when substituted, substituent group can be substituted on any workable tie point, and the substituent group is preferably independently optionally chosen from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, nitro, cyano, naphthenic base, heterocycle, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkylthio group, C (O) OR ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸In one or more substituent groups replaced.

Term " naphthenic base " refers to that the unsaturated monocycle of saturation or part or polycyclic cyclic hydrocarbon substituent, cycloalkyl ring include 3 to 20 carbon atoms, preferably comprises 3 to 12 carbon atoms, includes more preferably 3 to 10 carbon atoms, most preferably comprises 3 to 6 carbon atoms.The non-limiting example of monocyclic cycloalkyl includes cyclopropyl, cyclobutyl, cyclopenta, cyclopentenyl, cyclohexyl, cyclohexenyl group, cyclohexadienyl, suberyl, cycloheptatriene base, cyclooctyl etc., preferably cyclopropyl, cyclobutyl, cyclopenta and cyclohexyl；Polycyclic naphthene base includes the naphthenic base of loop coil, condensed ring and bridged ring.

Term " spiro cycloalkyl group " refers to the polycyclic moiety that a carbon atom (claiming spiro-atom) is shared between 5 to 20 yuan of monocycle, can be containing one or more double bonds, but none ring has the pi-electron system of total conjugated.Preferably 6 to 14 yuan, more preferably 7 to 10 yuan.Spiro cycloalkyl group is divided into single spiro cycloalkyl group, double spiro cycloalkyl groups or more spiro cycloalkyl groups according to the number for sharing spiro-atom between ring and ring, preferably single spiro cycloalkyl group and double spiro cycloalkyl groups.More preferably 4 yuan/4 yuan, 4 yuan/5 yuan, 4 yuan/6 yuan, 5 yuan/5 yuan or 5 yuan/6 yuan single spiro cycloalkyl groups.The non-limiting example of spiro cycloalkyl group includes:

Term " cycloalkyl " refers to 5 to 20 yuan, the full carbon polycyclic moiety of shared a pair of of the carbon atom adjoined of other rings in each ring and system in system, wherein one or more rings can be containing one or more double bonds, but none ring has the pi-electron system of total conjugated.Preferably 6 to 14 yuan, more preferably 7 to 10 yuan.Bicyclic, tricyclic, Fourth Ring or polycyclic fused ring alkyl, preferably bicyclic or tricyclic, more preferably 5 yuan/5 yuan or 5 yuan/6 membered bicyclic alkyl can be divided into according to a group cyclic number.The non-limiting example of cycloalkyl includes:

With

Term " bridge ring alkyl " refers to 5 to 20 yuan, and any two ring shares the full carbon polycyclic moiety of two carbon atoms being not directly connected, can be containing one or more double bonds, but none ring has the pi-electron system of total conjugated.Preferably 6 to 14 yuan, more preferably 7 to 10 yuan.Bicyclic, tricyclic, Fourth Ring or polycyclic bridge ring alkyl, preferably bicyclic, tricyclic or Fourth Ring can be divided into according to a group cyclic number, be more selected as bicyclic or tricyclic.The non-limiting example of bridge ring alkyl includes:

With

The cycloalkyl ring can be condensed on aryl, heteroaryl or heterocycloalkyl ring, wherein the ring to link together with precursor structure is naphthenic base, non-limiting example includes indanyl, tetralyl, benzocyclohepta alkyl etc..Naphthenic base can be optionally replacing or non-substituted, when substituted, substituent group is preferably independently optionally chosen from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, nitro, cyano, naphthenic base, Heterocyclylalkyl, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkylthio group, C (O) OR ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸In one or more substituent groups replaced.

Term " heterocycle " refers to the full and/or unsaturated monocycle in part or polycyclic cyclic hydrocarbon substituent, and it includes 3 to 20 annular atoms, wherein one or more annular atoms are selected from nitrogen, oxygen or S (O) _mThe hetero atom of (wherein m is integer 0 to 2), but do not include the loop section of-O-O- ,-O-S- or-S-S-, remaining annular atom is carbon.3 to 12 annular atoms are preferably comprised, wherein 1~4 is hetero atom；More preferable heterocycle includes 3 to 10 annular atoms, and most preferably heterocycle includes 3 to 6 annular atoms.The non-limiting example of monocyclic heterocycles base include oxetanylmethoxy, azelidinyl, tetrahydrofuran base, THP trtrahydropyranyl, pyrrole radicals, piperidyl, piperazinyl, morpholinyl, thio-morpholinyl, high piperazine base and Deng preferably azelidinyl, oxetanylmethoxy, pyrrole radicals and piperidyl；Multiring heterocyclic includes the heterocycle of loop coil, condensed ring and bridged ring.

Term " spiro heterocyclic radical " refers to that the polycyclic heterocyclic group that an atom (claiming spiro-atom) is shared between 5 to 20 yuan of monocycle, wherein one or more annular atoms are selected from nitrogen, oxygen or S (O) _mThe hetero atom of (wherein m is integer 0 to 2), remaining annular atom are carbon.It can be containing one or more double bonds, but none ring has the pi-electron system of total conjugated.Preferably 6 to 14 yuan, more preferably 7 to 10 yuan.Spiro heterocyclic radical is divided into single spiro heterocyclic radical, double spiro heterocyclic radicals or more spiro heterocyclic radicals according to the number for sharing spiro-atom between ring and ring, preferably single spiro cycloalkyl group and double spiro cycloalkyl groups.More preferably 4 yuan/4 yuan, 4 yuan/5 yuan, 4 yuan/6 yuan, 5 yuan/5 yuan or 5 yuan/6 yuan single spiro heterocyclic radicals.The non-limiting example of spiro heterocyclic radical includes:

Term " condensed hetero ring base " refers to 5 to 20 yuan, the polycyclic heterocyclic group of shared a pair of of the atom adjoined of other rings in each ring and system in system, one or more rings can contain one or more double bonds, but none ring has the pi-electron system of total conjugated, wherein one or more annular atoms are selected from nitrogen, oxygen or S (O) _mThe hetero atom of (wherein m is integer 0 to 2), remaining annular atom are carbon.Preferably 6 to 14 yuan, more preferably 7 to 10 yuan.Bicyclic, tricyclic, Fourth Ring or polycyclic condensed hetero ring base, preferably bicyclic or tricyclic, more preferably 5 yuan/5 yuan or 5 yuan/6 membered bicyclic condensed hetero ring bases can be divided into according to a group cyclic number.The non-limiting example of condensed hetero ring base includes:

With

Term " bridge heterocycle " refers to 5 to 14 yuan, any two ring shares the polycyclic heterocyclic group of two atoms being not directly connected, it can contain one or more double bonds, but none ring has the pi-electron system of total conjugated, wherein one or more annular atoms are selected from nitrogen, oxygen or S (O) _mThe hetero atom of (wherein m is integer 0 to 2), remaining annular atom are carbon.Preferably 6 to 14 yuan, more preferably 7 to 10 yuan.7 to 10 yuan.Bicyclic, tricyclic, Fourth Ring or polycyclic bridge heterocycle, preferably bicyclic, tricyclic or Fourth Ring, more preferably bicyclic or tricyclic can be divided into according to a group cyclic number.The non-limiting example of bridge heterocycle includes:

With

The heterocyclic ring can be condensed on aryl, heteroaryl or cycloalkyl ring, wherein the ring to link together with precursor structure is heterocycle, non-limiting example includes:

With

Heterocycle can be optionally replacing or non-substituted, when substituted, substituent group is preferably one or more following groups, is independently optionally chosen from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, nitro, cyano, naphthenic base, Heterocyclylalkyl, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkylthio group, C (O) OR ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸In one or more substituent groups replaced.

Term " aryl " refers to 6 to the 14 yuan of full carbon monocycles or fused polycycle (rings of namely shared adjacent carbon atoms pair) group, preferably 6 to 10 yuan of the pi-electron system with conjugation, such as phenyl and naphthalene.The aryl rings can be condensed on heteroaryl, heterocycle or cycloalkyl ring, wherein the ring to link together with precursor structure is aryl rings, non-limiting example includes:

Aryl can be substituted or non-substituted, when substituted, substituent group is preferably one or more following groups, is independently optionally chosen from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, nitro, cyano, naphthenic base, Heterocyclylalkyl, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkylthio group, C (O) OR ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸In one or more substituent groups replaced.

Term " heteroaryl " refers to the heteroaromatic system comprising 1 to 4 hetero atom, 5 to 14 annular atoms, and wherein hetero atom is selected from oxygen, sulphur and nitrogen.Heteroaryl is preferably 5 to 10 yuan, more preferably 5- or 6-membered, such as furyl, thienyl, pyridyl group, pyrrole radicals, N- alkyl pyrrole radicals, pyrimidine radicals, pyrazinyl, imidazole radicals, pyrazolyl, tetrazole radical etc..The heteroaryl ring can be condensed on aryl, heterocycle or cycloalkyl ring, wherein the ring to link together with precursor structure is heteroaryl ring, non-limiting example includes:

With

Heteroaryl can be optionally replacing or non-substituted, when substituted, substituent group is preferably one or more following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, nitro, cyano, naphthenic base, Heterocyclylalkyl, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkylthio group, C (O) OR ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸In one or more substituent groups replaced.

Term " alkoxy " refers to-O- (alkyl) and-O- (non-substituted naphthenic base), and wherein alkyl is as defined above.The non-limiting example of alkoxy includes: methoxyl group, ethyoxyl, propoxyl group, butoxy, cyclopropyl oxygroup, cyclobutoxy group, cyclopentyloxy, cyclohexyloxy.Alkoxy can be optionally replacing or non-substituted, when substituted, substituent group is preferably one or more following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio group, alkyl amino, halogen, sulfydryl, hydroxyl, amino, nitro, cyano, naphthenic base, Heterocyclylalkyl, aryl, heteroaryl, cycloalkyloxy, heterocyclylalkoxy groups, cycloalkylthio, heterocycle alkylthio group, C (O) OR ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸In one or more substituent groups replaced.

Term " hydroxyalkyl " refers to the alkyl being optionally substituted by a hydroxyl group, and wherein alkyl is as defined above.

Term " halogenated alkyl " refers to that alkyl is replaced by one or more halogens, and wherein alkyl is as defined above.

Term " hydroxyl " refers to-OH group.

Term " sulfydryl " refers to-SH group.

Term " halogen " refers to fluorine, chlorine, bromine or iodine.

Term " amino " refers to-NH ₂。

Term " cyano " refers to-CN.

Term " nitro " refers to-NO ₂。

Term " oxo base " refers to=O.

" optional " or " optionally " mean event or environment described later can with but need not occur, which includes that the event or environment occur or not spot occasion.For example, mean " optionally by alkyl-substituted heterocyclic group " alkyl can with but necessarily exist, the explanation include heterocyclic group by alkyl-substituted situation and heterocyclic group not by alkyl-substituted situation.

" substituted " refers to that one or more hydrogen atoms in group, preferably at most 5, more preferably 1~3 hydrogen atom are replaced by the substituent group of respective number independently of one another.Self-evident, substituent group is only in their possible chemistry position, and those skilled in the art can determine in the case where not paying and excessively making great efforts and (pass through experiment or theory) possible or impossible substitution.It may be unstable when for example, amino or hydroxyl with free hydrogen are in conjunction with the carbon atom with unsaturated (such as olefinic) key.

" pharmaceutical composition " indicates mixture and other components such as physiology/pharmaceutical carrier and excipient containing one or more compounds described herein or its physiologically/pharmaceutical salt or pro-drug and other chemical constituents.The purpose of pharmaceutical composition is the administration promoted to organism, plays bioactivity in turn conducive to the absorption of active constituent.

" officinal salt " refers to the salt of the compounds of this invention, has safety and validity when this kind of salt is used in the mammalian body, and have due bioactivity.

M and R ⁶~R ⁸As defined in logical formula (I).

The synthetic method of the compounds of this invention

In order to complete the purpose of the present invention, the present invention adopts the following technical scheme:

Scheme one

The present invention lead to formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt preparation method, comprising the following steps:

The first step, compound and the halogenating agent reaction of general formula (I-1), obtains the compound of general formula (I-A)；

Second step, the compound reaction of the compound and general formula (I-3) of general formula (I-2), obtains the compound of general formula (I-4)；

Third step, the compound and NHR of general formula (I-4) ⁴R ⁵Compound reaction, obtain the compound of general formula (I-Bb)；

4th step, under alkaline condition, in the presence of a catalyst, reaction obtains the compound of general formula (I-B) to the compound and boranes compound of general formula (I-Bb)；

The compound of 5th step, the compound of general formula (I-A) and general formula (I-B) under alkaline condition, is reacted through Suzuki in the presence of a catalyst, obtains the compound of logical formula (I)；R ¹When for THP trtrahydropyranyl, it can slough in acid condition；

The reagent for providing alkaline condition includes organic base and inorganic base, the organic bases include but is not limited to triethylamine, N, N- diisopropylethylamine, n-BuLi, lithium diisopropylamine, lithium hexamethyldisilazide, potassium acetate, sodium tert-butoxide or potassium tert-butoxide, the inorganic base include but is not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide；

There is provided acid condition reagent include but is not limited to hydrogen chloride, the 1,4- dioxane solution of hydrogen chloride, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid, phosphoric acid, to benzene methanesulfonic acid, Me ₃SiCl and TMSOT _f；

Halogenating agent includes but is not limited to bromine, hydrogen bromide, N- bromo-succinimide (NBS), PBr ₃、POBr ₃, cross pyridinium bromide hydrobromide (PHP), tetrabromo cyclic ketones (TBCO), bromo diethyl malonate, tetrabutylammonium bromide, N-chlorosuccinimide, PCl ₃And POCl ₃；

Catalyst includes but is not limited to palladium/carbon, Raney's nickel, tetra-triphenylphosphine palladium, palladium chloride, palladium acetate, [1, bis- (diphenylphosphino) ferrocene of 1'-] palladium chloride, 1,1 '-bis- (dibenzyl phosphorus) dichloro diamyl iron palladiums, tris(dibenzylideneacetone) dipalladium or 2- dicyclohexyl phosphine -2', 6'- dimethoxy-biphenyl, preferably [1, bis- (diphenylphosphino) ferrocene of 1'-] palladium chloride or 2- dicyclohexyl phosphine -2', 6'- dimethoxy-biphenyl；

Boranes compound includes but is not limited to 4,4,5,5- tetramethyl -1,3,2- dioxaborolan, 4,4,4', 4', 5,5,5', 5'- prestox -2,2'- two (1,3,2- dioxaborolanes), connection boric acid neopentyl glycol ester, B (OB _u-n) ₃Or B (OP _r-i) ₃；

Above-mentioned reaction preferably carries out in a solvent, solvent for use includes but is not limited to: acetic acid, methanol, ethyl alcohol, toluene, tetrahydrofuran, methylene chloride, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1,4- dioxane, water, n,N-Dimethylformamide and its mixture；

Wherein:

W is Or

X is halogen, preferably bromine；

R ⁹For alkyl；Preferably C _1-6Alkyl；

Ring A, R ¹~R ⁵, n and s be as defined in logical formula (I).

Scheme two

The present invention lead to formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt preparation method, comprising the following steps:

The first step, compound and the halogenating agent reaction of general formula (I-1), obtains the compound of general formula (I-A)；

Second step, under alkaline condition, in the presence of a catalyst, reaction obtains the compound of general formula (I-Aa) to the compound and boranes compound of general formula (I-A)；

Third step, the compound reaction of the compound and general formula (I-3) of general formula (I-2), obtains the compound of general formula (I-4)；

4th step, the compound and NHR of general formula (I-4) ⁴R ⁵Compound reaction, obtain the compound of general formula (I-Bb)；

The compound of 5th step, the compound of general formula (I-Bb) and general formula (I-Aa) under alkaline condition, is reacted through Suzuki in the presence of a catalyst, obtains the compound of logical formula (I)；R ¹When for THP trtrahydropyranyl, it can slough in acid condition；

The reagent for providing alkaline condition includes organic base and inorganic base, the organic bases include but is not limited to triethylamine, N, N- diisopropylethylamine, n-BuLi, lithium diisopropylamine, lithium hexamethyldisilazide, potassium acetate, sodium tert-butoxide or potassium tert-butoxide, the inorganic base include but is not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide；

There is provided acid condition reagent include but is not limited to hydrogen chloride, the 1,4- dioxane solution of hydrogen chloride, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid, phosphoric acid, to benzene methanesulfonic acid, Me ₃SiCl and TMSOT _f；

Halogenating agent includes but is not limited to bromine, hydrogen bromide, N- bromo-succinimide (NBS), PBr ₃、POBr ₃, cross pyridinium bromide hydrobromide (PHP), tetrabromo cyclic ketones (TBCO), bromo diethyl malonate, tetrabutylammonium bromide, N-chlorosuccinimide, PCl ₃And POCl ₃；

Boranes compound includes but is not limited to 4,4,5,5- tetramethyl -1,3,2- dioxaborolan, 4,4,4', 4', 5,5,5', 5'- prestox -2,2'- two (1,3,2- dioxaborolanes), connection boric acid neopentyl glycol ester, B (OB _u-n) ₃Or B (OP _r-i) ₃；

Catalyst includes but is not limited to palladium/carbon, Raney's nickel, tetra-triphenylphosphine palladium, palladium chloride, palladium acetate, [1, bis- (diphenylphosphino) ferrocene of 1'-] palladium chloride, 1,1 '-bis- (dibenzyl phosphorus) dichloro diamyl iron palladiums or tris(dibenzylideneacetone) dipalladium, preferably [1,1'- bis- (diphenylphosphino) ferrocene] palladium chloride；

Above-mentioned reaction preferably carries out in a solvent, solvent for use includes but is not limited to: acetic acid, methanol, ethyl alcohol, toluene, tetrahydrofuran, methylene chloride, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1,4- dioxane, water, n,N-Dimethylformamide and its mixture；

Wherein:

W is Or

X is halogen, preferably bromine；

R ⁹For alkyl；Preferably C _1-6Alkyl；

Ring A, R ¹~R ⁵, n and s be as defined in logical formula (I).

Specific embodiment

Embodiment

The structure of compound is by nuclear magnetic resonance (NMR) or/and mass spectrum (MS) come what is determined.NMR is displaced (δ) with 10 ^-6(ppm) unit provides.The measurement of NMR is to use Bruker AVANCE-400 nuclear magnetic resonance spectrometer, and measurement solvent is deuterated dimethyl sulfoxide (DMSO-d ₆), deuterated chloroform (CDCl ₃), deuterated methanol (CD ₃OD), inside it is designated as tetramethylsilane (TMS).

The measurement of MS is with FINNIGAN LCQAd (ESI) mass spectrograph (manufacturer: Thermo, model: Finnigan LCQ advantage MAX).

The measurement of HPLC uses Agilent 1200DAD high pressure liquid chromatograph (Sunfire 150 × 4.6mm of C18 chromatographic column) and Waters 2695-2996 high pressure liquid chromatograph (Gimini 150 × 4.6mm of C18 chromatographic column).

Chiral HPLC measurement uses LC-10A vp (Shimadzu) or SFC-analytical (Berger Instruments Inc.)；

Tlc silica gel plate uses Yantai Huanghai Sea HSGF254 or Qingdao GF254 silica gel plate, and the specification that the silica gel plate that thin-layered chromatography (TLC) uses uses is 0.15mm~0.2mm, and the specification that thin-layer chromatography isolates and purifies product use is 0.4mm~0.5mm.

Column chromatography is generally carrier using 200~300 mesh silica gel of Yantai Huanghai Sea silica gel.

Chiral preparatory column chromatography uses Prep Star SD-1 (Varian Instruments Inc.) or SFC-multigram (Berger Instruments Inc.).

The quick preparing instrument of CombiFlash uses Combiflash Rf200 (TELEDYNE ISCO).

Kinases average inhibition and IC ₅₀The measurement of value is with NovoStar microplate reader (German BMG company).

Known starting material of the invention can be used or be synthesized according to methods known in the art, or it is commercially available from ABCR GmbH & Co.KG, Acros Organics, Aldrich Chemical Company, splendid remote chemistry scientific and technological (Accela ChemBio Inc) reach the companies such as auspicious chemicals.

Without specified otherwise in embodiment, reaction can be carried out under argon atmospher or nitrogen atmosphere.

Argon atmospher or nitrogen atmosphere refer to that reaction flask connects the argon gas or nitrogen balloon of an about 1L volume.

Nitrogen atmosphere refers to that reaction flask connects the hydrogen balloon of an about 1L volume.

Pressure hydration reaction hydrogenates instrument using Parr 3916EKX type hydrogenation instrument and clear indigo plant QL-500 type hydrogen generator or HC2-SS type.

Hydrogenation usually vacuumizes, and is filled with hydrogen, operates 3 times repeatedly.

Microwave reaction uses 908860 type microwave reactor of CEM Discover-S.

Without specified otherwise in embodiment, solution refers to aqueous solution.

Without specified otherwise in embodiment, it is 20 DEG C~30 DEG C that the temperature of reaction, which is room temperature,.

The monitoring of reaction process in embodiment uses thin-layered chromatography (TLC), solvent used in reacting, the system of eluant, eluent and the solvent system of thin-layered chromatography for the column chromatography that purifying compound uses include: A: methylene chloride/methanol system, B: n-hexane/ethyl acetate system, C: petrol ether/ethyl acetate system, D: petrol ether/ethyl acetate/methanol, the volume ratio of solvent is different according to the polarity of compound and is adjusted, and the alkalinity such as a small amount of triethylamine and acetic acid can also be added or acid reagent is adjusted.

Embodiment 1

6- (1- cyclopropyl -3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 1

The first step

2- (1- cyclopropyl -1H- pyrazole-3-yl) -6- picoline 1c

By 2- methyl -6- (1H- pyrazole-3-yl) pyridine 1a (700mg, 4.40mmol, using well known method " Bioorganic and Medicinal Chemistry; 2015; 23 (6); 1260-1275 " is prepared), cyclopropylboronic acid 1b (756mg, 8.80mmol), copper acetate dihydrate (1.76g, 8.80mmol), sodium carbonate (933mg, 8.80mmol) and 2,2- bipyridyl (1.37g, it 8.80mmol) is dissolved in 25mL 1, in 2- dichloroethanes, is warming up to 50 DEG C and is stirred to react 16 hours.After reaction, filter, filtrate decompression concentration, with silica gel column chromatography with eluant, eluent system B purify obtained by residue, obtain title compound 1c (440mg, yield: 50.2%).

MS m/z(ESI):200.2[M+1]

Second step

2- (the bromo- 1- cyclopropyl -1H- pyrazole-3-yl of 4-) -6- picoline 1d

Compound 1c (330mg, 1.66mmol) is dissolved in 8mL methylene chloride, is added N- bromo-succinimide (295mg, 1.66mmol), is stirred at room temperature 1 hour.After reaction, water is added in reaction solution, methylene chloride extracts (10mL × 3), merge organic phase, be concentrated under reduced pressure, with silica gel column chromatography with eluant, eluent system A purify obtained by residue, obtain title compound 1d (335mg, yield: 72.6%).

Third step

2- (1- cyclopropyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base) -1H- pyrazole-3-yl) -6- picoline 1f

By compound 1d (400mg, 1.44mmol), 4,4,5,5- tetramethyls -1,3,2- dioxaborolan 1e (1.84g, 14.4mmol), 2- dicyclohexyl phosphine -2', 6'- dimethoxy-biphenyl (89mg, 0.216mmol), triethylamine (364mg, 3.6mmol) and two (cyano benzene) palladium chloride (18.6mg, 0.072mmol) it is dissolved in toluene, under the conditions of 90 DEG C, microwave reaction 1 hour.Add water in reaction solution, be extracted with ethyl acetate (30mL × 2), merge organic phase, anhydrous sodium sulfate is dry, filtrate, filtrate decompression concentration are collected in filtering, residue silica gel column chromatography obtains title compound 1f (300mg, yield: 64.1%) with eluant, eluent system A purifying.

MS m/z(ESI):244.4[M-82+1]

4th step

Bromo- [1,2,4] triazol [4,3-a] the pyridine-3-carboxylic acid ethyl ester 1h of 6-

Under argon atmospher, by bromo- 2- hydrazino pyridine 1g (4g, the 21.27mmol of 5-, be prepared using method disclosed in patent application " US20140134133 "), 2- ethyl (2.17g, 21.27mmol) be dissolved in 60mL methanol, at 60 DEG C, react 1h, reacting liquid temperature is down to room temperature, is concentrated under reduced pressure, and 60mL 1 is added into concentrate, 4- dioxane, it is added iodobenzene diethylester (8.22g, 25.53mmol), reacts 18 hours.It is concentrated under reduced pressure, mixes sample and cross column, residue silica gel chromatography obtains title compound 1h (4.5g, yield: 70.49%) with eluant, eluent system D purifying.

MS m/z(ESI):270.3[M+1]

5th step

Bromo- [1,2,4] triazol [4,3-a] the pyridine-3-carboxamide 1i of 6-

Compound 1h (500mg, 1.85mmol) is dissolved in 7N methanolic ammonia solution (8.37mL, 58.58mmol), is stirred to react 1 hour.Reaction solution is concentrated under reduced pressure, and residue silica gel chromatography obtains title compound 1i (450mg, yield: 99%) with eluant, eluent system A purifying.

MS m/z(ESI):241.3[M+1]

6th step

6- (1- cyclopropyl -3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 1

Under argon atmospher, successively by compound 1f (75mg, 0.23mmol), compound 1i (55.59mg, 0.23mmol), double Diphenyl phosphino ferrocene (12.79mg, 0.02mmol), [1,1'- bis- (diphenylphosphino) ferrocene] palladium chloride (16.87mg, 0.02mmol) and potassium carbonate (63.75mg, 0.46mmol) it is dissolved in the mixed solution (V/V=5:1) of 12mL Isosorbide-5-Nitrae-dioxane and water.Under the conditions of 80 DEG C, it is stirred to react 18 hours.Reaction solution is cooled to room temperature, filtering, and filtrate decompression concentration, residue silica gel column chromatography obtains title compound 1 (35mg, yield: 40%) with eluant, eluent system I purifying.

MS m/z(ESI):360.4[M+1]

¹H NMR(400MHz,CD ₃OD)δ9.32(s,1H),8.13(s,1H),7.79-7.72(m,2H),7.61-7.56(m,2H),7.24-7.22(d,1H),3.83-3.79(m,1H),2.40(s,3H),1.28-1.23(m,2H),1.15-1.10(m,2H).

Embodiment 2

6- (1- cyclobutyl -3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 2

The first step

(3- carbamoyl-[1,2,4] triazol [4,3-a] pyridine -6- base) boric acid 2a

Compound 1i (4.2g is sequentially added under an argon, 17.42mmol), 4, 4, 4', 4', 5, 5, 5', 5'- prestox -2, 2'- bis- (1, 3, 2- dioxaborolanes) (6.64g, 26.14mmol, using well known method " Journal of the American Chemical Society, 2009, 131 (5), 1656-1657 " is prepared), [1, bis- (diphenylphosphino) ferrocene of 1'-] palladium chloride (1.29g, 1.74mmol) and potassium acetate (4.28g, 43.56mmol) it is dissolved in 80mL1, in 4- dioxane solution, it is heated to 80 DEG C, it stirs Mix reaction 3 hours.Stop reaction, reaction solution is concentrated under reduced pressure, and residue silica gel column chromatography obtains title compound 2a (2.6g, yield: 36.2%) with eluant, eluent system A purifying.

MS m/z(ESI):207.4[M+1]

Second step

2- (1- cyclobutyl -1H- pyrazole-3-yl) -6- picoline 2c

Successively by compound 1a (2.01g, 12.6mmol), 4- toluenesulfonic acid ring butyl ester 2b (4.29g, 18.9mmol, using well known method " Journal of the American Chemical Society; 1980; 102 (11), 3863-3870 " are prepared) and 100mL N is added in cesium carbonate (8.23g, 25.3mmol), in dinethylformamide, it finishes, under the conditions of 60 DEG C, is stirred to react 12 hours.Reaction solution is cooled to room temperature, water is added, is extracted with ethyl acetate (20mL × 3), merges organic phase, (10mL × 2) are washed with saturated sodium chloride solution, anhydrous sodium sulfate dries, filters, and collects filtrate, filtrate decompression concentration, residue silica gel column chromatography obtains title compound 2c (1.81g, yield: 67.3%) with eluant, eluent system B purifying.

MS m/z(ESI):214.4[M+1]

Third step

2- (the bromo- 1- cyclobutyl -1H- pyrazole-3-yl of 4-) -6- picoline 2d

Compound 2c (1.81g, 8.49mmol) is dissolved in 18mL methylene chloride, is added N- bromo-succinimide (1.51g, 8.49mmol), is stirred at room temperature 1 hour.After reaction, reaction solution be concentrated under reduced pressure, with silica gel column chromatography with eluant, eluent system B purify obtained by residue, obtain title compound 2d (2.13g, yield: 85.9%).

MS m/z(ESI):292.3[M+1]

4th step

6- (1- cyclobutyl -3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 2

Under argon atmospher, successively by compound 2a (78.25mg, 0.38mmol), compound 2d (74mg, 0.25mmol), potassium carbonate (70.01mg, 0.51mmol), double Diphenyl phosphino ferrocenes (14.04mg, 0.03mmol) and [1,1'- bis- (diphenylphosphino) ferrocene] palladium chloride (18.53mg, 0.03mmol) it is dissolved in the mixed solution (V/V=10:1) of 11mL Isosorbide-5-Nitrae-dioxane and water.Under the conditions of 80 DEG C, it is stirred to react 12 hours.Reaction solution is cooled to room temperature, and 20mL water is added in reaction solution, then (10mL × 3) are extracted with ethyl acetate, merge organic phase, (10mL × 2) are washed with saturated sodium chloride solution, anhydrous sodium sulfate dries, filters, collect filtrate, filtrate decompression concentration, residue silica gel column chromatography are purified with eluant, eluent system A purifying, gained crude product with high performance liquid chromatography, obtain title compound 2 (4mg, yield: 4.2%).

MS m/z(ESI):374.5[M+1]

¹H NMR(400MHz,CDCl ₃)δ9.34(s,1H),7.76(d,1H),7.72(s,1H),7.64-7.62(m,2H),7.54(d,1H),7.42(brs,1H),7.11(d,1H),5.76(brs,1H),4.93-4.84(m,1H),2.66-2.56(m,4H),2.46(s,3H),1.96-1.88(m,2H).

Embodiment 3

6- (1- methyl -3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 3

The first step

2- methyl -6- (1- methyl-1 H- pyrazole-3-yl) pyridine 3a

Compound 1a (1g, 6.28mmol), iodomethane (1.07g, 7.54mmol) and potassium carbonate (1.74g, 12.56mmol) are added in 10mL n,N-Dimethylformamide, reacted 12 hours.After reaction, reaction solution is concentrated under reduced pressure, and residue silica gel column chromatography obtains title compound 3a (320mg, yield: 29.4%) with eluant, eluent system A purifying.

MS m/z(ESI):174.4[M+1]

Second step

2- (the bromo- 1- methyl-1 H- pyrazole-3-yl of 4-) -6- picoline 3b

Compound 3a (240mg, 1.39mmol) and N- bromo-succinimide (294.29mg, 1.66mmol) are added in 10mL methylene chloride, are stirred to react 12 hours.After reaction, 10mL saturated sodium bicarbonate solution is added into reaction solution, then is extracted with ethyl acetate, organic phase is collected, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, obtains crude title compound 3b (349mg), and product is directly used in without further purification to react in next step.

MS m/z(ESI):253.1[M+1]

Third step

6- (1- methyl -3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 3 is under argon atmospher, successively by crude product 3b (300mg, 1.19mmol), compound 2a (294.11mg, 1.43mmol), potassium carbonate (328.92mg, 2.38mmol) and [1, bis- (diphenylphosphino) ferrocene of 1'-] palladium chloride (88.27mg, 0.119mmol) it is dissolved in the mixed solution (V/V=50:1) of 10.2mL Isosorbide-5-Nitrae-dioxane and water.Under the conditions of 90 DEG C, it is stirred to react 12 hours.Diatomite filtering, filtrate decompression concentration, residue are purified with high performance liquid chromatography, obtain title compound 3 (120mg, yield: 30.3%).

MS m/z(ESI):334.4[M+1]

¹H NMR(400MHz,CDCl ₃)δ9.39(s,1H),7.80(d,1H),7.67(s,1H),7.63(t,1H),7.58-7.56(dd,2H),7.40(s,1H),7.12(d,1H),5.71(s,1H),4.08(s,3H),2.47(s,3H).

Embodiment 4

6- (1- ethyl -3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 4

The first step

2- (1- ethyl -1H- pyrazole-3-yl) -6- picoline 4a

Compound 1a (1.2g, 7.54mmol), iodoethane (1.18g, 7.54mmol) and potassium carbonate (2.08g, 15.08mmol) are added in 5mL n,N-Dimethylformamide, reacted 12 hours.After reaction, reaction solution is concentrated under reduced pressure, and residue silica gel column chromatography obtains title compound 4a (520mg, yield: 36.8%) with eluant, eluent system A purifying.

MS m/z(ESI):188.4[M+1]

Second step

2- (the bromo- 1- ethyl -1H- pyrazole-3-yl of 4-) -6- picoline 4b

Compound 4a (460mg, 2.46mmol) and N- bromo-succinimide (521.80mg, 2.95mmol) are added in 10mL methylene chloride, are stirred to react 12 hours.After reaction, 10mL saturated sodium bicarbonate solution is added into reaction solution, then is extracted with ethyl acetate, organic phase is collected, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, obtains crude title compound 4b (650mg), and product is directly used in without further purification to react in next step.

MS m/z(ESI):267.2[M+1]

Third step

6- (1- ethyl -3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 4 is under argon atmospher, successively by crude product 4b (300mg, 1.13mmol), compound 2a (278.61mg, 1.35mmol), potassium carbonate (311.58mg, 2.25mmol) and [1, bis- (diphenylphosphino) ferrocene of 1'-] palladium chloride (83.62mg, 0.112mmol)) it is dissolved in the mixed solution of 10.2mL Isosorbide-5-Nitrae-dioxane and water (V/V=50:1).Under the conditions of 90 DEG C, it is stirred to react 12 hours.Diatomite filtering, filtrate decompression concentration, residue are purified with high performance liquid chromatography, obtain title compound 4 (125mg, yield: 31.9%).

MS m/z(ESI):348.4[M+1]

¹H NMR(400MHz,CDCl ₃)δ9.39(s,1H),7.79(d,1H),7.70(s,1H),7.63(t,1H),7.59-7.56(dd,2H),7.39(s,1H),7.12(d,1H),5.68(s,1H),4.34(q,2H),2.46(s,3H),1.65(t,3).

Embodiment 5

6- (1- isopropyl -3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 5

The first step

2- (1- isopropyl -1H- pyrazole-3-yl) -6- picoline 5a

Compound 1a (1.2g, 7.54mmol), 2- N-Propyl Bromide (927.14mg, 7.54mmol) and potassium carbonate (2.08g, 15.08mmol) are added in 5mL n,N-Dimethylformamide, reacted 12 hours.After reaction, reaction solution is concentrated under reduced pressure, and residue silica gel column chromatography obtains title compound 5a (520mg, yield: 36.8%) with eluant, eluent system A purifying.

MS m/z(ESI):202.4[M+1]

Second step

2- (the bromo- 1- isopropyl -1H- pyrazole-3-yl of 4-) -6- picoline 5b

Compound 5a (250mg, 1.24mmol) and N- bromo-succinimide (263.82mg, 1.49mmol) are added in 10mL methylene chloride, are stirred to react 12 hours.Reaction terminates, and reaction solution is concentrated under reduced pressure, and residue silica gel chromatography obtains title compound 5b (348mg, yield: 100%) with eluant, eluent system A purifying.

MS m/z(ESI):281.2[M+1]

Third step

6- (1- isopropyl -3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 5

Under argon atmospher, successively by crude product 5b (300mg, 1.07mmol), compound 2a (330.82mg, 1.61mmol), potassium carbonate (295.99mg, 2.14mmol) and [1, bis- (diphenylphosphino) ferrocene of 1'-] palladium chloride (79.43mg, 107.08 μm of ol) is dissolved in the mixed solution (V/V=50:1) of 10.2mL Isosorbide-5-Nitrae-dioxane and water.Under the conditions of 90 DEG C, it is stirred to react 12 hours.Diatomite filtering, filtrate decompression concentration, residue are purified with high performance liquid chromatography, obtain title compound 5 (132 mg, yield: 34.1%).

MS m/z(ESI):362.5[M+1]

¹H NMR(400MHz,CDCl ₃)δ9.39(t,1H),7.80-7.78(dd,1H),7.71(s,1H),7.64(s,1H),7.63(d,1H),7.59-7.56(d,1H),7.41(s,1H),7.12-7.10(m,1H),5.69(s,1H),4.71-4.64(m,1H),2.45(s,3H),1.64(d,6H).

Embodiment 6

6- (3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 6

The first step

2- (the bromo- 1H- pyrazole-3-yl of 4-) -6- picoline 6a

Compound 1a (1g, 6.28mmol) and N- bromo-succinimide (1.12g, 6.28mmol) are added in 30mL methylene chloride, are stirred to react 1.5 hours.Reaction terminates, 10mL unsaturated carbonate potassium solution is added into reaction solution, (10mL × 3) are extracted with dichloromethane again, merge organic phase, organic phase is dried, filtered with anhydrous sodium sulfate, filtrate decompression concentration, residue silica gel column chromatography obtains title compound 6a (1.45g, yield: 87.3%) with eluant, eluent system A purifying.

MS m/z(ESI):238.5[M+1]

Second step

2- (the bromo- 1- of 4- (tetrahydro -2H- pyrans -2- base) -1H- pyrazole-3-yl) -6- picoline 6b

Under argon atmospher, by compound 6a (1.4g, 5.88mmol) and 3,4- dihydro -2H- pyrans (1.12g, it 6.28mmol) is added in 20mL toluene, then above-mentioned reaction solution is added in trifluoroacetic acid (0.03g, 0.29mmol), under the conditions of 80 DEG C, it is stirred to react 48 hours.Reaction terminates, 10mL saturated sodium carbonate solution is added into reaction solution, (20mL × 3) are extracted with ethyl acetate again, merge organic phase, organic phase is dried, filtered with anhydrous sodium sulfate, filtrate decompression concentration, residue silica gel column chromatography obtains title compound 6b (1.45g, yield: 87.26%) with eluant, eluent system C purifying.

MS m/z(ESI):322.5[M+1]

Third step

6- (3- (6- picoline -2- base) -1- (tetrahydro -2H- pyrans -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 6c

Under argon atmospher, successively by compound 6b (300mg, 0.93mmol), compound 2a (9.59mg, 0.05mmol), potassium carbonate (386.06mg, 2.79mmol) and [1, bis- (diphenylphosphino) ferrocene of 1'-] palladium chloride (690.67mg, 0.93mmol) is dissolved in the mixed solution of 16.5mL Isosorbide-5-Nitrae-dioxane and water (V/V=10:1).Under the conditions of 80 DEG C, it is stirred to react 18 hours.Ethyl acetate is added, organic phase is collected in washing, and anhydrous sodium sulfate dries, filters, and filtrate decompression concentration, residue is purified with high performance liquid chromatography, obtains title compound 6c (150mg, yield: 35.94%).

MS m/z(ESI):404.5[M+1]

4th step

6- (3- (6- picoline -2- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 6

Under argon atmospher, compound 6c (20mg, 0.05mmol) is added in 2mL Isosorbide-5-Nitrae-dioxane, under the conditions of 0 DEG C, 4mL 4N hydrogen chloride Isosorbide-5-Nitrae-dioxane solution is added dropwise in Xiang Shangshu reaction solution, finishes, react 2 hours.Reaction solution is concentrated under reduced pressure, and residue is purified with high performance liquid chromatography, obtains title compound 6 (5mg, yield: 26.9%).

MS m/z(ESI):427.2[M+1]

¹H NMR(400MHz,CD ₃OD)δ9.38(s,1H),8.03-8.07(m,1H),7.83-7.85(m,1H),7.74-7.78(m,1H),7.64-7.67(m,1H),7.49-7.54(m,1H),7.27-7.29(m,1H),3.50(m,1H),3.15(m,1H),2.48(s,3H)

Embodiment 7

(R) -6- (3- (6- picoline -2- base) -1- (tetrahydrofuran -3- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 7

The first step

(R) -2- methyl -6- (1- (tetrahydrofuran -3- base) -1H- pyrazole-3-yl) pyridine 7b

By compound 1a (637mg, 4.0mmol) it is dissolved in 20mL N, in dinethylformamide, (S)-tetrahydrofuran -3- base 4- oluene sulfonic acides ester 7a (1.45g is added, 6.0mmol, it is prepared using method disclosed in patent application " WO2014049133 ") and cesium carbonate (2.61g, 8.0mmol), 60 DEG C are stirred to react 16 hours.Reaction solution be concentrated under reduced pressure, with the quick preparing instrument of CombiFlash with eluant, eluent system B purify obtained by residue, obtain title compound 7b (610mg, yield: 66.5%).

MS m/z(ESI):230.4[M+1]

Second step

(R) -2- (the bromo- 1- of 4- (tetrahydrofuran -3- base) -1H- pyrazole-3-yl) -6- picoline 7c

Compound 7b (600mg, 2.62mmol) is dissolved in 30mL methylene chloride, is added N- bromo-succinimide (466mg, 2.62mmol), is stirred at room temperature 16 hours.After reaction, be concentrated under reduced pressure, with the quick preparing instrument of CombiFlash with eluant, eluent system B purify obtained by residue, obtain title compound 7c (795mg, yield: 98.6%).

MS m/z(ESI):310.3[M+1]

Third step

(R) -6- (3- (6- picoline -2- base) -1- (tetrahydrofuran -3- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 7

Under argon atmospher, successively by compound 7c (115mg, 0.37mmol), compound 2a (307.45mg, 1.49mmol), double Diphenyl phosphino ferrocene (20.69mg, 0.04mmol), [1, bis- (diphenylphosphino) ferrocene of 1'-] palladium chloride (27.31mg, 0.040mmol) and potassium carbonate (103.15mg, 0.75mmol) it is dissolved in the mixed solution of 16.5mL Isosorbide-5-Nitrae-dioxane and water (V/V=10:1).Under the conditions of 80 DEG C, it is stirred to react 18 hours.Reaction solution is cooled to room temperature, and 30mL water is added in reaction solution, then (20mL × 3) are extracted with ethyl acetate, merge organic phase, (10mL × 2) are washed with saturated sodium chloride solution, anhydrous sodium sulfate dries, filters, collect filtrate, filtrate decompression concentration, residue silica gel chromatography are purified with eluant, eluent system A purifying, gained crude product with high performance liquid chromatography, obtain title compound 7 (8mg, yield: 5.5%).

MS m/z(ESI):390.4[M+1]

¹H NMR(400MHz,CDCl ₃)δ9.31(s,1H),7.82-7.77(m,3H),7.58(d,1H),7.51(d,1H),7.35(brs,1H),7.26(s,1H),5.64(brs,1H),5.17-5.12(m,1H),4.28-4.22(m,2H),4.14-4.10(m,1H),4.02-3.97(m,1H),2.63(s,3H),2.61-2.54(m,2H).

Embodiment 8

(S) -6- (3- (6- picoline -2- base) -1- (tetrahydrofuran -3- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 8

The first step

(S) -2- methyl -6- (1- (tetrahydrofuran -3- base) -1H- pyrazole-3-yl) pyridine 8b

By compound 1a (328mg, 2.06mmol) it is dissolved in 10mL N, in dinethylformamide, (R)-tetrahydrofuran -3- base 4- oluene sulfonic acides ester 8a (500mg is added, 2.06mmol, it is prepared using method disclosed in patent application " WO2016021192 ") and cesium carbonate (1.3g, 4.12mmol), 60 DEG C are stirred to react 2 hours.Reaction solution is concentrated under reduced pressure, and water is added in residue, and ethyl acetate extraction merges organic phase, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, residue silica gel column chromatography purifies gained residue with eluant, eluent system A, obtains title compound 8b (175mg, yield: 37.0%).

MS m/z(ESI):230.4[M+1]

Second step

(S) -2- (the bromo- 1- of 4- (tetrahydrofuran -3- base) -1H- pyrazole-3-yl) -6- picoline 8c

Compound 8b (370mg, 1.6mmol) is dissolved in 18mL methylene chloride, is added N- bromo-succinimide (341.76mg, 1.92mmol), is stirred at room temperature 12 hours.After reaction, water is added in reaction solution, methylene chloride extraction merges organic phase, anhydrous sodium sulfate dries, filters, filtrate decompression concentration, residue silica gel column chromatography purifies gained residue with eluant, eluent system A, obtains title compound 8c (325.5mg, yield: 65.1%).

Third step

(S) -6- (3- (6- picoline -2- base) -1- (tetrahydrofuran -3- base) -1H- pyrazoles -4- base)-[1,2,4] triazol [4,3-a] pyridine-3-carboxamide 8

Under argon atmospher, successively by compound 8c (100mg, 0.32mmol), compound 2a (300mg, 1.46mmol), double Diphenyl phosphino ferrocene ((17.99mg, 0.0300mmol), [1, bis- (diphenylphosphino) ferrocene of 1'-] palladium chloride (23.74mg, 0.03mmol) and potassium carbonate (89.7mg, 0.65mmol) it is dissolved in the mixed solution of 11mL Isosorbide-5-Nitrae-dioxane and water (V/V=10:1).Under the conditions of 85 DEG C, it is stirred to react 12 hours.Reaction solution is cooled to room temperature, 30mL water is added in reaction solution, then (10mL × 3) are extracted with ethyl acetate, merges organic phase, anhydrous sodium sulfate is dry, filtrate, filtrate decompression concentration are collected in filtering, residue silica gel chromatography is with eluant, eluent system A purifying, gained crude product is purified with high performance liquid chromatography, obtains title compound 8 (14mg, yield: 11.1%).

MS m/z(ESI):390.4[M+1]

¹H NMR(400MHz,CDCl ₃)δ9.39(s,1H),7.81(d,1H),7.78(s,1H),7.62(d,1H),7.57(d,1H),7.42(s,1H),7.14(d,1H),5.75-5.69(m,1H),5.19-5.13(m,1H),4.29-4.23(m,2H),4.18-4.12(m,1H),4.06-4.00(m,1H),2.66-2.55(m,1H),2.48(s,3H).

Test case:

Biological assessment

The measurement of test case 1, the compounds of this invention to the inhibiting effect of TGF β RI (ALK5) kinase activity

The inhibiting effect of external TGF β RI (ALK5) kinase activity is tested by the following method.

The compounds of this invention uses following determination of experimental method to the TGF β active inhibiting effect of RI kinases ALK5:

Enzyme assay uses TGF β RI kinase assay kit (V4093, Promega), and 2 μ l reaction buffer (40mM Tris pH7.5,20mM MgCl is sequentially added in 384 orifice plates (4514, Corning) ₂0.1mg/ml BSA) prepare enzyme solutions (the final concentration of 2ng/ μ L of enzyme in reaction system), 1 μ l is dissolved in the compound of 3 times of gradient dilutions of 5%DMSO, (final concentration of 50 μM of ATP of mixed solution of 2 μ l ATP and TGF β RI substrate polypeptides, Final substrate concentrations are 0.2 μ g/ μ L), after 27 DEG C are reacted 2.5 hours, the ADP-Glo solution in 5 μ l kits is added in every hole, 27 DEG C are placed 40 minutes, every hole adds 10 μ l kinase assay reagents, and 27 DEG C are placed 30 minutes.Chemiluminescence signal value is detected using (PerkinElmer) multi-function microplate reader of Victor 3.Compound is calculated to the IC of enzyme inhibition with corresponding signal value according to each concentration of compound with Graphpad prism software ₅₀Value.

The bioactivity of the compounds of this invention is measured by above test, the IC measured ₅₀Value see the table below 1.

IC of 1 the compounds of this invention of table to TGF β RI kinases ALK5 activity suppression ₅₀

Embodiment number	IC 50(nM)
1	80
3	426
4	426
5	181

6

27

Conclusion: compound of the embodiment of the present invention has significantly inhibiting effect to TGF β RI kinases ALK5 activity.

The measurement of test case 2, the compounds of this invention to the inhibiting effect of VEGFR2 kinase activity

The inhibiting effect of external VEGFR2 kinase activity is tested by the following method.

Experimental method as described below is used to measure the compounds of this invention to the inhibiting effect of VEGFR2 kinase activity:

Enzyme assay uses 1 Peptide (PV3190 of Kinase Assay Kit-Tyrosine, Invitrogen) kit, 5 μ l reaction buffer (50mM HEPES pH7.5,10mM MgCl is sequentially added in 384 orifice plates (4513, Corning) ₂1mM EGTA, 0.05% BRIJ-35) prepare recombinant human VEGF R2 enzyme (PV3660,) and VEGFR2 substrate polypeptide (the final concentration of 0.14ng/ μ L of enzyme in reaction system Invitrogen, Final substrate concentrations are 2 μM), 2.5 μ l are dissolved in the compound of 2 times of gradient dilutions of 5%DMSO, 2.5 μ L ATP solution (final concentration of 50 μM of ATP), after 25 DEG C are reacted 2 hours, 5 μ L detection reagents are added in every hole, after 25 DEG C are placed 1 hour, with the fluorescence signal value of NOVOstar (BMG) multi-function microplate reader detection launch wavelength 445nm and 520nm.Compound is calculated to the IC of enzyme inhibition with corresponding signal value according to each concentration of compound with Graphpad prism software ₅₀Value.

The bioactivity of the compounds of this invention is measured by above test, the IC measured ₅₀Value see the table below 2.

IC of 2 the compounds of this invention of table to the inhibiting effect of VEGFR2 kinase activity ₅₀

Embodiment number	IC 50(nM)
1	>10000
6	1001

Conclusion: compound of the embodiment of the present invention is weak to VEGFR2 kinase activity inhibiting effect, illustrates that compound of the embodiment of the present invention has selective inhibitory to TGF β RI kinases.

The measurement of test case 3, the compounds of this invention to the inhibiting effect of p 38 alpha kinase activity

The inhibition of external p 38 alpha kinase activity is tested by the following method.

Experimental method as described below is used to measure the compounds of this invention to the inhibiting effect of p 38 alpha kinase activity:

Enzyme assay uses p 38 alpha kinase assay kit (V9591, Promega), and 2 μ L reaction buffer (40mM Tris pH7.5,20mM MgCl is sequentially added in 384 orifice plates (4514, Corning) ₂0.1mg/mL BSA) prepare enzyme solutions (the final concentration of 0.5ng/ μ L of enzyme in reaction system), 1 μ L is dissolved in the compound of 3 times of gradient dilutions of 5%DMSO, the mixed solution (final concentration of 50 μM of ATP, Final substrate concentrations are 0.2 μ g/ μ L) of 2 μ L ATP and p38 substrate polypeptides, after 27 DEG C are reacted 2.5 hours, the ADP-Glo solution in 5 μ L kits is added in every hole, 27 DEG C are placed 40 minutes, and every hole adds 10 μ L kinase assay reagents, and 27 DEG C are placed 30 minutes.Chemiluminescence signal value is detected using (PerkinElmer) multi-function microplate reader of Victor 3.Compound is calculated to the IC of enzyme inhibition with corresponding signal value according to each concentration of compound with Graphpad prism software ₅₀Value.

The bioactivity of the compounds of this invention is measured by above test, the IC measured ₅₀Value see the table below 3.

IC of 3 the compounds of this invention of table to the inhibiting effect of p 38 alpha kinase activity ₅₀

Embodiment number	IC 50(nM)
1	1368
6	2613

Conclusion: compound of the embodiment of the present invention is weak to the inhibiting effect of p 38 alpha kinase activity, illustrates that compound of the embodiment of the present invention has selective inhibitory to TGF β RI kinases.

Test case 4, the compounds of this invention measure the inhibition of NIH3T3 cell Proliferation

Following in vitro test is for measuring the compounds of this invention to the inhibitory activity of NIH3T3 cell Proliferation.

Experimental method as described below is used to measure the compounds of this invention to the inhibiting effect of NIH3T3 cell Proliferation:

In 96 hole clear bottom blanks (3903, Corning with the DMEM culture medium (SH30243.01 containing 10%FBS in), GE) every hole is inoculated with 100 μ L NIH3T3 cell (GNM6, the American Type Culture Collection committee, Chinese Academy of Sciences cell bank), inoculum density is 2000 cells/wells, cell is at 37 DEG C, 5%CO ₂Under the conditions of overnight incubation.After being incubated overnight, every hole is changed to DMEM culture medium of the 90 μ L containing 0.5%FBS, and the 10 μ L compound of 3 times of gradient dilutions of DMEM culture medium containing 0.5%FBS is then added, and places 37 DEG C, 5%CO ₂It is cultivated 72 hours in cell incubator.50 μ L CellTiter-Glo (G7573, Promega) are added in last every hole, and incubation at room temperature reads chemiluminescence signal value using Victor3 microplate reader (PerkinElmer) after ten minutes.Calculate the IC of compound with corresponding signal value according to each concentration of compound with Graphpad Prism software ₅₀Value.

The compounds of this invention bioactivity from the above analysis gained, calculate resulting IC ₅₀Value such as the following table 4:

IC of 4 the compounds of this invention of table to the inhibition of NIH3T3 cell Proliferation ₅₀

Embodiment number	IC 50(nM)
1	246
2	357
5	203
6	40

Conclusion: the compounds of this invention has apparent inhibitory activity to NIH3T3 cell Proliferation.

The measurement of test case 5, the compounds of this invention to the inhibitory activity of the Smad signal path of TGF β RI

Following in vitro test is for measuring the compounds of this invention to the inhibitory activity of the Smad signal path of TGF β RI.

Experimental method as described below is used to measure the compounds of this invention to the inhibitory activity of the Smad signal path of TGF β RI:

It is inoculated with 100 μ L HepG2 (TCHu 72, the American Type Culture Collection committee, Chinese Academy of Sciences cell bank) cell with the every hole of EMEM culture solution (42360-099, Gibco) containing 10%FBS in 96 orifice plates, inoculum density is 2.5 × 10 ⁴Cells/well, cell is at 37 DEG C, 5%CO ₂Under the conditions of overnight incubation.The EMEM fresh medium containing 10%FBS is replaced, every hole transfects 0.1 μ g 3TP-lux plasmid (11767, general such as spit of fland biotechnology (Beijing) Co., Ltd), and cell continues at 37 DEG C, 5%CO ₂Under the conditions of cultivate 24 hours.EMEM culture solution of the 90 μ L containing 0.5%FBS is replaced in every hole, 6 hours hungry.Compound is configured to the storing liquid of 20mM, with 100%DMSO gradient dilution at 400 × concentration, then with the EMEM containing 0.5%FBS dilute 40 times.Tissue culture plate is taken out, every hole is separately added into compound or control (0.25%DMSO) after 10 μ L dilution, and gently oscillation mixes, and places 37 DEG C, 5%CO ₂It is cultivated 18 hours in incubator, 100 μ L detection reagent ONE-Glo are added in last every hole ^TMLuciferase Assay (E6110, Promega) room temperature avoid light place 10 minutes, reads chemiluminescence signal value using Victor3.0 (PerkinElmer).Calculate the IC of compound with corresponding signal value according to each concentration of compound with Graphpad Prism software ₅₀Value.

The compounds of this invention bioactivity from the above analysis gained, calculate resulting IC ₅₀Value such as the following table 5:

The IC that 5 the compounds of this invention of table inhibits the Smad signal path of TGF β RI ₅₀

Embodiment number	IC 50(nM)
1	167
2	40
6	34

Conclusion: the compounds of this invention has apparent inhibitory activity to the Smad signal path of TGF β RI.

Pharmacokinetic Evaluation

The pharmacokinetics test of test case 6, the compounds of this invention

1, it makes a summary

Using rat as animal subject, rat oral gavage is determined using LC/MS/MS method and gives 1 compound of embodiment, the drug concentration in rear different moments blood plasma.The compounds of this invention is studied in the intracorporal pharmacokinetics behavior of rat, evaluates its characteristics of pharmacokinetics.

2, testing program

2.1 test drug

1 compound of embodiment.

2.2 experimental animal

Healthy adult SD rat 4, half male and half female is purchased from Shanghai Jie Sijie experimental animal Co., Ltd, animal productiong licensing number: SCXK (Shanghai) 2013-0006.

2.3 drugs are prepared

A certain amount of drug is weighed, adds the normal saline of the DMSO of 5% volume, the Tween 80 of 5% volume and 90% volume at the achromaticity and clarification transparency liquid of 0.2mg/mL.

2.4 administration

Gastric infusion after SD Rat Fast is stayed overnight, dosage is 2.0mg/kg, and administered volume is 10.0mL/kg.

3. operation

1 compound of embodiment is administered in rat oral gavage, before administration and 0.5,1.0,2.0,4.0 after administration, it is taken a blood sample 0.2mL by eye socket within 6.0,8.0,11.0,24.0 hours, it is placed in heparinised tubes, 4 DEG C, 3500 revs/min of centrifugations, 10 minutes separated plasmas, is saved in -20 DEG C, fed within 2 hours after administration.

Untested compound content after the drug gastric infusion of measurement various concentration in rat plasma: the 25 μ L of rat plasma at each moment after medicine is drawn, 80 μ L (100ng/mL) of inner mark solution camptothecine is added, 200 μ L of acetonitrile, vortex mixed 5 minutes, 10 minutes (4000 revs/min) are centrifuged, plasma sample takes 1.0 μ L of supernatant to carry out LC/MS/MS analysis.

4, pharmacokinetic parameter result

The pharmacokinetic parameter of the compounds of this invention is as follows:

Conclusion: in the medicine generation of the compounds of this invention, absorbs preferably, has pharmacokinetic advantage.

Claims (16)

1. A kind of logical formula (I) compound represented:

   Or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt,

   Wherein:

   Ring A is aryl or heteroaryl；

   R ¹Selected from hydrogen atom, alkyl, halogenated alkyl, hydroxyl, hydroxyalkyl, amino, naphthenic base, heterocycle, aryl, heteroaryl ,-C (O) OR ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸, wherein alkyl, naphthenic base, heterocycle, aryl and the heteroaryl are optionally selected from halogen, alkyl, alkoxy, halogenated alkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, naphthenic base, heterocycle, aryl, heteroaryl ,-C (O) OR each independently ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸In one or more substituent groups replaced；

   R ²It is identical or different, and it is each independently selected from hydrogen atom, halogen, alkyl, alkoxy, halogenated alkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, naphthenic base, heterocycle, aryl, heteroaryl ,-C (O) OR ⁶、-C(O)R ⁶、-S(O) _mR ⁶、-NR ⁷R ⁸、-S(O) _mNR ⁷R ⁸With-C (O) NR ⁷R ⁸；

   R ³It is identical or different, and it is each independently selected from hydrogen atom, halogen, alkyl, alkoxy, halogenated alkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, naphthenic base, heterocycle, aryl and heteroaryl；

   R ⁴And R ⁵It is each independently selected from hydrogen atom, alkyl, halogenated alkyl, naphthenic base, heterocycle, aryl and heteroaryl, wherein alkyl, naphthenic base, heterocycle, aryl and the heteroaryl are each independently optionally replaced one or more substituent groups in alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, naphthenic base, heterocycle, aryl and heteroaryl；

   Alternatively, the R ⁴And R ⁵Heterocycle is formed together with the nitrogen-atoms being connected, wherein in the heterocycle containing 1~2 identical or different hetero atom selected from N, O and S, and the heterocycle is optionally replaced one or more substituent groups in alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, naphthenic base, heterocycle, aryl and heteroaryl；

   R ⁶Selected from hydrogen atom, alkyl, amino, halogenated alkyl, naphthenic base, heterocycle, aryl and heteroaryl；

   R ⁷And R ⁸It is each independently selected from hydrogen atom, alkyl, halogenated alkyl, naphthenic base, heterocycle, aryl and heteroaryl, wherein alkyl, naphthenic base, heterocycle, aryl and the heteroaryl are each independently optionally replaced one or more substituent groups in alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, naphthenic base, heterocycle, aryl and heteroaryl；

   Alternatively, the R ⁷And R ⁸Heterocycle is formed together with the nitrogen-atoms being connected, wherein in the heterocycle containing 1~2 identical or different hetero atom selected from N, O and S, and the heterocycle is optionally replaced one or more substituent groups in alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, naphthenic base, heterocycle, aryl and heteroaryl；

   N is 0,1,2,3 or 4；

   S is 0,1,2 or 3；And

   M is 0,1 or 2.
2. Logical formula (I) compound represented according to claim 1, middle ring A are heteroaryl, preferably 5- or 6-membered heteroaryl, more preferable pyridyl group.
3. Logical formula (I) compound represented according to any one of claim 1 or 2, to lead to formula (II) compound represented:

   Or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt,

   Wherein: R ¹~R ⁵, n and s it is as defined in claim 1.
4. Logical formula (I) compound represented described in any one of claim 1 to 3, wherein R ⁴And R ⁵It is hydrogen atom.
5. Logical formula (I) compound represented according to any one of claims 1 to 4, to lead to formula (III) compound represented:

   Or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt,

   Wherein: R ¹And R ²As defined in claim 1.
6. Logical formula (I) compound represented according to any one of claims 1 to 5, wherein R ¹Selected from hydrogen atom, alkyl, naphthenic base and heterocycle.
7. Logical formula (I) compound represented described according to claim 1~any one of 6, wherein R ²For hydrogen atom or alkyl.
8. Logical formula (I) compound represented according to any one of claims 1 to 4, wherein R ³For hydrogen atom.
9. According to claim 1, logical formula (I) compound represented described in~any one of 8, be selected from:
10. A method of logical formula (I) compound represented according to claim 1 is prepared, this method comprises:

    The compound reaction of the compound and general formula (I-B) of general formula (I-A), obtains the compound of logical formula (I),

    Wherein:

    W is boronate or 4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base；

    X is halogen, preferably bromine；

    Ring A, R ¹~R ⁵, n and s it is as defined in claim 1.
11. A method of logical formula (I) compound represented according to claim 1 is prepared, this method comprises:

    The compound reaction of the compound and general formula (I-Bb) of general formula (I-Aa), obtains the compound of logical formula (I),

    Wherein:

    W is boronate or 4,4,5,5- tetramethyl -1,3,2- dioxaborolan -2- base；

    X is halogen, preferably bromine；

    Ring A, R ¹~R ⁵, n and s it is as defined in claim 1.
12. A kind of pharmaceutical composition, described pharmaceutical composition contain therapeutically effective amount according to claim 1~any one of 9 described in logical formula (I) compound represented or its tautomer, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt and one or more pharmaceutically acceptable carriers, diluent or excipient.
13. Logical formula (I) compound represented or its tautomer described according to claim 1~any one of 9, mesomer, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt or pharmaceutical composition according to claim 12 preparing the purposes in the drug for treating, preventing or reduce Nasopharyngeal neoplasms.
14. Logical formula (I) compound represented or its tautomer described according to claim 1~any one of 9, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form or its pharmaceutical salt or pharmaceutical composition according to claim 12 in preparation for treating, preventing or reducing the purposes in the drug by the tumour of TGF-β overexpression mediation.
15. Logical formula (I) compound represented or its tautomer described according to claim 1~any one of 9, racemic modification, enantiomter, diastereoisomer, or mixtures thereof form, or its pharmaceutical salt or pharmaceutical composition according to claim 12 are being prepared for treating, purposes in the drug of prevention or mitigation disease, the disease is selected from: cardiovascular disease, all kinds of inflammation, tumour, the fibrosis of the various causes of disease, injury of blood vessel, nephrosis, hepatosis, tuberculosis, adult respiratory distress syndrome (ARDS), intimal thickening, eye disease, arteriopathy or hypertrophica corium scar or keloid during the wound healing caused by wound or wound occurs to be formed, peritonaeum and sub-dermal adhesion, chorionitis, fibrosclerosis, progressive systemic sclerosis, osteoporosis, ulcer, nervous function subtracts Low, male erectile dysfunction, Peyronie's disease, dupuytren's contracture, Alzheimer's disease and Raynaud's syndrome.
16. Logical formula (I) compound represented or its tautomer described according to claim 1~any one of 9, racemic modification, enantiomter, diastereoisomer, or mixtures thereof the purposes of form or its pharmaceutical salt or pharmaceutical composition according to claim 12 in the drug that preparation inhibits TGF-β signal transduction path.