CN110294761A

CN110294761A - Substituted pyrazolo [1,5-a] pyrimidine compound as Trk kinase inhibitor

Info

Publication number: CN110294761A
Application number: CN201810586447.XA
Authority: CN
Inventors: 范晶晶; 唐春雷; 范为正
Original assignee: Nanjing Lei Zheng Medicine Technology Co Ltd
Current assignee: Nanjing Lei Zheng Medicine Technology Co Ltd
Priority date: 2018-06-08
Filing date: 2018-06-08
Publication date: 2019-10-01
Anticipated expiration: 2038-06-08
Also published as: CN110294761B

Abstract

The present invention relates to a kind of substituted pyrazolo [1,5-a] pyrimidine compounds or its pharmaceutically acceptable salt, pharmaceutical composition and purposes as Trk kinase inhibitor.Pyrazolo [1,5-a] pyrimidine derivatives with logical formula (I) structure have good Trk family protein tyrosine-kinase enzyme inhibition activity and metabolic stability.

Description

Substituted pyrazolo [1,5-a] pyrimidine compound as Trk kinase inhibitor

Technical field

The invention belongs to chemical medicine, the present invention relates to a series of substituted pyrazolo [1,5-a] pyrimidine compounds, The purposes of pharmaceutical composition comprising the compound and the compound.

Background technique

Tropomyosin associated kinase (tropomyosin-related kinase, Trk) is a kind of nerve growth factor Receptor, family have high homology tropomyosin associated kinase A (tropomyosin-related kinase A, TrkA), tropomyosin associated kinase B (TrkB), tropomyosin associated kinase C (TrkC) form, respectively by NTRK1, NTRK2 and NTRK3 coding.There is (i) that the nerve growth factor (NGF) of TrkA can be activated in neurotrophic factor, (ii) can be activated The brain-derived neurotrophic factor (BDNF) and NT-4/5 of TrkB, and (iii) can activate the NT3 of TrkC.Trk finds expression in mind extensively Through member tissue in and (Current Opinion in related with the maintenance of neuronal cell, signal transduction and survival Neurobiology,2001,11:272-280)。

It is related that Trk kinases is initially considered growth, differentiation, apoptosis to neuronal cell etc., Trk/ neurotrophic factor The inhibitor of approach is effective in zootype before the various clinical of pain.For example, the way Antagonism TrkA/NGF has been displayed Diameter antibody (for example, RN-624) is effective in inflammatory and neuropathic pain zootype and in human clinical trial (Neuroscience,1994,62:327-331；J.Pain,2004,5:157-163；Nat.Med.1995,1:774-780； Neuroreport,1997,8:807-810).In addition, recently document show, after inflammation, in dorsal root ganglion BDNF content and TrkB signal transduction will increase (Brain Research, 1997,749:358), and several researches show that make by BDNF/TrkB The antibody that the signal transduction of approach reduces can inhibit the effect of neuron allergy and related pain (Molecular Pain, 2008,4: 27).The performance of crossing that Trk also has been displayed in nearest document reaches, activates, expands and/or is mutated, these cancer packets related with many cancers It includes melanoma (Journal of Investigative Dermatology, 2008,128 (8): 2031-2040), it is non-small thin Born of the same parents' lung cancer (Molecular and Cellular Biochemistry, 2007,295 (1&2): 19-26), thyroid cancer (Neuroendocrinology Letters, 2007,28 (3): 221-229), acute myeloid leukemia (Cancer Lett, 2001,169:107-14), glioblastoma (Journal of Neurochemistry, 2007,103:259-275), star Cytoma and medulloblastoma (Brain Pathology, 2006,16:304-310), colon cancer (Science, 2003,300: 949), neuroblastoma (Nat.Rev.Cancer, 2003,3:203-216), oophoroma (Clin.Cancer Res.2003, 9:2248-2259), breast cancer (Brain Pathology, 2006,16:304-310), prostate cancer (Clin.Cancer Res.1998,4 (8): 1887-1898), cancer of pancreas (Journal ofGastroenterology and Hepatology, 2006,21 (5): 850-858), Huppert's disease (Cancer Genetics and Cytogenetics, 2007,178:1- And maxicell neuroendocrine tumors (Human Mutation, 2008,29 (5): 609-616) 10).In the preclinical mould of cancer In formula, the non-selective micromolecular inhibitor of Trk A, B and C and Trk/Fc chimera is inhibiting tumour growth and is preventing tumour Transfer aspect effectively (Cancer Letters, 2001,169:107-114；Cancer Letters,2006,232:90-98； Cancer Res,2008,68:(2)346-351).In cancer clinical trials, small molecule Trk kinase inhibitor Larotrectinib (LOXO-101) treats LMNA-NTRK1 Gene Fusion type soft tissue sarcoma transfer patient, four In week, lung's sarcoma volume obviously becomes smaller, and the transfer and diffusion of cancer cell slow down, while patient respiratory difficulty is delayed Solution, after four months, pulmonary masses disappear substantially, and obvious adverse reaction (Cancer is not found during clinical treatment Discov,2015,5:1049-1057).LOXO-101 is on carrying the positive adult/pediatric cancer patient of NTRK1/2/3 fusion With significant curative effect, such as soft tissue malignant tumour, thyroid cancer, salivary-gland carcinoma, gastrointestinal stromal tumor, children's fibrosarcoma, non- Small cell carcinoma, cholangiocarcinoma, kidney, salivary-gland carcinoma, appendix cancer, cancer of pancreas, Peripheral Nerve Sheath Tumors, spindle cell tumor etc., and the medicine Object Small side effects (New England Journal of Medicine, 2018,378 (8): 731-739.).Trk kinase inhibition The reality that agent Entrectinib (NMS-P626) resets 18 with NTRK1/2/3, ROS1 or ALK gene in clinical test Body tumor patient carries out clinical experimental study, the results showed that 72% patient has reaction (J Target to Entrectinib Ther Cancer,2015,12:34-37)。

In addition, inhibition neurotrophic factor/Trk approach has been displayed can effectively treat inflammatory disease in clinical premode Disease.For example, inhibit neurotrophic factor/Trk approach related with the clinical premode of following diseases: including asthma Inflammatory lung disease (Pharmacology&Therapeutics, 2008,117 (1): 52-76), interstitial cystitis (The Journal of Urology, 2005,173 (3): 1016-1021) including ulcerative colitis and Crohn disease (Crohn's Disease inflammatory bowel disease (Gut, 2000,46 (5): 670-678) and inflammatory dermatosis including), such as atopic dermatitis (Archives of Dermatological Research, 2006,298 (1): 31-37), eczema and chronic eczema (Journal of Investigative Dermatology,2004,122(3):812-819)。

Neurotrophic factor/Trk approach, specifically BDNF/TrkB approach also has with the etiology of neurodegenerative disease It closes, these diseases include multiple sclerosis, Parkinson's disease (Parkinson's disease) and A Zihaimoshi disease (Alzheimer's disease)(Frontiers in Neuroendocrinology,2006,27(4):404-414).It adjusts Section neurotrophic factor/Trk approach can be used for treating these diseases and related disease.

Research shows that TrkA receptor is for schizotrypanum cruzi (Typanosoma cruzi) (Cha Jiasishi disease (Chagasdisease)) most important (the Cell Host& of lysis in the infection of the parasitic infection in human host Microbe,2007,1(4):251-261).Therefore, TrkA is inhibited to can be used for treating Cha Jiasishi disease and relevant protozoan infection.

Meanwhile Trk inhibitor also can be used for treating related with bone remoulding Imbalance disease, such as osteoporosis, Rheumatoid arthritis and Bone tumour.Bone tumour is the frequent complication of cancer, in advanced breast cancer or patients with prostate cancer Up to 70% and lung cancer, colon cancer, gastric cancer, bladder cancer, uterine cancer, the carcinoma of the rectum, thyroid cancer or patients with renal cell carcinoma in about 15 to 30% can occur the complication.Ostelytic metastases can cause severe pain, pathologic fracture, the hypercalcinemia of threat to life, ridge Marrow stress disorders and other nerve compression syndromes wait group.Therefore, the drug that can induce proliferative TNF-a Induced Apoptosis in Osteoblasts will be extremely advantageous.? Performance (Bone, 2000,26 (6): 625- of TrkA receptor and TrkC receptor are observed in the skeletonization region of fracture mouse model 633).In addition, observing the distribution of NGF in almost all of osteoblast.Pan-Trk inhibitor can mankind hFOB at In osteocyte, the Tyrosine signal transduction that is activated after inhibiting by neurotrophic factor in conjunction with all 3 Trk receptors.These Data are supported to use the theory of Trk inhibitor for treating bone remoulding disease (such as Bone tumour in cancer patient).

Micromolecular inhibitor (the Expert of known several Trk kinases that can be used for cancer or pain Opin.Ther.Patents,2009,19(3):305-319)。

If international patent application WO2006115452 and WO2006087538 elaborate that Ganlei is stated to be inhibitor or Trk swashs The small molecule of enzyme can be used for treating pain or cancer.

International patent application WO2007025540 discloses certain imidazo [1,2-b] pyridazines being substituted, and has at 6 There is the piperazinyl of secondary amino group or BOC- protection.These compounds are the inhibitor for being disclosed as protein kinase C (PKC).

International patent application WO2008052734 discloses (R) -4- (6- (2- (3- fluorophenyl) Pyrrolizidine -1- base) imidazoles And [1,2-b] pyridazine -3- base) benzonitrile, that is, imidazo [1,2b] pyridazine compound, in 6 heterocycles substituted with aryl - Group and 3 have benzonitrile group.It is said that the compound is receptor-mediated by PI3K receptor, JAK-2 receptor and Trk suitable for treating Disease.

International patent application WO2007013673 discloses 1- phenyl -3- (6- (1- phenylethylcarbamate) imidazo [1,2- B] pyridazine -3- base) urea and N- (6- (4- hydroxy-cyclohexyl amino) imidazo [1,2-b] pyridazine -3- base) benzamide, that is, miaow Azoles simultaneously [1,2b] pyridazine compound has amide or urea part with amino and at 3 at 6.These compounds are stated to be Trk inhibitor.Novel and significantly more efficient treatment is continuously needed to mitigate pain, especially chronic ache.Due to TrkA and its Its Trk kinases can be used as the mediator of the biological respinse of NGF driving, therefore the inhibitor of TrkA and other Trk kinases can be chronic pain Pain state provides effectively treatment.

U.S. Patent Publication No. US20110195948 describes phonetic as the substituted pyrazolo [1,5-a] of Trk inhibitor Acridine compound.

In recent years, for curative effect more preferably, the Trk inhibitor that less side effects, metabolic stability is high, bioavilability is good deposits In urgent clinical demand.The present invention proposes a kind of novel substituted pyrazolecarboxylic as Trk kinase inhibitor simultaneously [1,5-a] pyrimidine Compound the deficiencies of to overcome the problems, such as stability of the existing technology, activity, meets growing clinical demand.

Summary of the invention

Problems to be solved by the invention

The present invention provides a kind of novel substituted pyrazolecarboxylic as Trk kinase inhibitor simultaneously [1,5-a] pyrimidine compound.It should Structural compounds pharmacodynamics performance is more preferable, metabolic stability is higher.

Further, the present invention provides a kind of compound with logical formula (I) or its pharmaceutically acceptable salt, medicine group Close object and purposes.

The solution to the problem

Present invention firstly provides a kind of compound with logical formula (I) or its pharmaceutically acceptable salt,

Wherein:

R¹It is (i) phenyl, is optionally independently selected from halogen, (1-4C) alkoxy, CF₃、CHF₂, one or more Substituent group replaces；Or (ii) 5 to 6 yuan of heteroaromatics, have selected from N and S heteroatom, wherein the heteroaromatic optionally by One or more halogen atoms replace；

R²Selected from NR^aR^b, (1-4C) alkyl, (1-4C) fluoroalkyl, (1-4C) hydroxy alkyl ,-(1-4C alkyl) miscellaneous Ar¹, it is miscellaneous Ar², miscellaneous Cyc¹, miscellaneous Cyc²Or it is optional by F, CF₃、OH、NH₂, OMe, OEt, NHMe replace (3-6C) naphthenic base；

R^aIt is H or (1-6C) alkyl；

R^bIt is H, (1-4C) alkyl, (1-4C) hydroxy alkyl, miscellaneous Ar³Or phenyl, wherein the phenyl is optionally independently selected From halogen, CF₃, CN one or more substituent groups replace；

Or NR^aR^b4 circle heterocyclic rings with theheterocyclic nitrogen atom are formed, wherein the heterocycle is optionally independently selected from below one Kind or multiple substituent groups replace: halogen, OH, (1-4C) alkyl, (1-4C) alkoxy；

Or NR^aR^b5 to 6 circle heterocyclic rings are formed, are formed with theheterocyclic nitrogen atom and optionally with selected from N, O and SO₂The second ring The heterocycle that hetero atom or group are formed replaces wherein the heterocycle is optionally independently selected from one or more substituent groups below: OH, halogen, CF₃, (1-4C) alkyl, NH₂,COOH；

Miscellaneous Ar¹It is 5 yuan of hetero-aromatic rings with 1 to 3 theheterocyclic nitrogen atom；

Miscellaneous Ar²It is 5 to 6 yuan of hetero-aromatic rings, is independently selected from N's and S wherein having at least one nitrogen ring atom and optionally having Second ring hetero atom, wherein the hetero-aromatic ring is optionally independently selected from one or more substituent groups of (1-4C) alkyl, halogen Replace；

Miscellaneous Cyc¹It is 4 to 6 yuan of azacyclo-s of carbon connection, is optionally independently selected from (1-4C) alkyl, CO₂(1-4C alkyl) One or more substituent groups replace；

Miscellaneous Cyc²It is the pyridone or pyridazine ring for being optionally selected from the substituent group of (1-4C) alkyl；

Miscellaneous Ar³It is 5 to 6 yuan of hetero-aromatic rings, is independently selected from the ring hetero atom of N and O with 1-2 and is optionally independently selected Replace from one or more substituent groups of (1-4C) alkyl；

X is selected from

R³Selected from OH, CN, (CH₂)_nOH、(CH₂)_nNH₂, wherein n is independent is expressed as 1,2 or 3；

Y is selected from O, NH or NR⁴；

R⁴Selected from (1-6C) alkyl.

Preferably, R²Selected from NR^aR^b, (1-4C) alkyl, (1-4C) fluoroalkyl, miscellaneous Ar², miscellaneous Cyc¹, miscellaneous Cyc².Further Preferably, R²Selected from NR^aR^b, miscellaneous Ar², wherein the NR^aR^bMiddle R^aSelected from H, R^bSelected from phenyl, phenyl is optionally by halogen, CF₃ One or more substituent groups replace or the NR^aR^b4 circle heterocyclic rings with theheterocyclic nitrogen atom are formed, wherein the heterocycle is optional Ground be independently selected from below a kind of or multiple substituent groups and replaced: halogen, OH, (1-4C) alkyl, (1-4C) alkoxy, or NR^aR^bFormed 5 to 6 circle heterocyclic rings, formed have theheterocyclic nitrogen atom heterocycle, the heterocycle be optionally independently selected from one below or Multiple substituent groups replace: OH, halogen, CF₃, (1-4C) alkyl, wherein the miscellaneous Ar²It is 6 yuan of hetero-aromatic rings, wherein there is at least one Theheterocyclic nitrogen atom and optionally has and be independently selected from the second ring hetero atom of N and S.

Preferably, R¹Selected from phenyl, it is optionally independently selected from halogen, CF₃、CHF₂One or more substituent groups take Generation.It is further preferred that R¹Selected from the phenyl replaced by one or two fluorine atom or chlorine atom or by a fluorine atom and CF₃Substituted phenyl.

Preferably, the compound is selected from:

Preferably, pharmaceutically acceptable salt of the present invention be inorganic salts or organic salt, inorganic salts include hydrochloride, Hydrobromate, hydriodate, sulfate, disulfate, nitrate, phosphate, acid phosphate；The organic salt is selected from acetic acid Salt, trifluoroacetate, propionate, acetonate, oxyacetate, oxalate, malonate, fumarate, maleate, cream Hydrochlorate, malate, citrate, tartrate, mesylate, with sulfonate, benzene sulfonate, salicylate.

The present invention also provides a kind of pharmaceutical composition, including the above-mentioned compound of the present invention or its is pharmaceutically acceptable Salt and pharmaceutically acceptable carrier, excipient or diluent.

In addition, the present invention provides compound of the present invention or its pharmaceutically acceptable salt in preparation for treating The cancer of mammal, pain, inflammation, neurodegenerative disease, infection by Trypanosoma cruzi or osteolytic disease drug in purposes. Preferably, wherein the cancer includes melanoma, non-small cell lung cancer, thyroid cancer, acute myeloid leukemia, pernicious glue Matter tumor, astrocytoma and medulloblastoma, colon cancer, neuroblastoma, oophoroma, breast cancer, prostate cancer, pancreas Cancer, Huppert's disease and maxicell neuroendocrine tumors；Wherein the pain be related to cancer, surgical operation, fracture, by Ostalgia caused by metastases, osteoarthritis, psoriatic arthritis, rheumatoid arthritis, interstitial cystitis, chronic pancreas Inflammation, splanchnodynia, inflammatory pain, migraine, chronic low back pain, painful bladder syndrome and neuropathic pain；The wherein inflammation Including asthma, interstitial cystitis, ulcerative colitis, Crohn disease, atopic dermatitis, eczema and chronic eczema.

The effect of invention

The substituted pyrazolecarboxylic with structure shown in logical formula (I) provided by the invention simultaneously [1,5-a] pyrimidine compound, has good Trk inhibitory activity and excellent pharmacodynamics performance, metabolic stability is higher and production cost is relatively low.

Specific embodiment

Technical solution of the present invention is described in detail below in conjunction with embodiment.

" (1-4C) alkyl " refers to the unit price of the linear chain or branched chain of the saturation of respectively 1 to 4 carbon atom in the present invention Alkyl.Example includes, but are not limited to methyl, ethyl, 1- propyl, 2- propyl, 1- butyl, 2- methyl-1-propyl, 2- butyl and 2- Methyl-2-propyl.

" (1-4C) alkoxy " refers to the list of the linear chain or branched chain of the saturation of respectively 1 to 4 carbon atom in the present invention Valence group, wherein the group is on oxygen atom.

" (1-4C) hydroxy alkyl " refers to the linear chain or branched chain of the saturation of respectively 1 to 4 carbon atom in the present invention Monovalent alkyl group, wherein 1 hydrogen atom is replaced by OH group.

The term " fluoroalkyl " being used in the present invention refers to the alkyl that one or more hydrogen atoms are replaced by fluorine atom.Fluorine The example of alkyl includes-CHF₂、–CH₂CF₃Or perfluoroalkyl such as-CF₃Or-CF₂CF₃。

It " is administered " in the present invention or " giving " individual compound is directed to individual in need for the treatment of and provides change of the invention Close object.

The present invention provides a kind of novel substituted pyrazolecarboxylic as Trk kinase inhibitor simultaneously [1,5-a] pyrimidine compounds Or its pharmaceutically acceptable salt, structural formula is as shown in logical formula (I):

Wherein:

R^aIt is H or (1-6C) alkyl, (1-6C) examples of alkyl includes methyl, ethyl, propyl, isopropyl, butyl, isobutyl Base；

R^bIt is H, (1-4C) alkyl, (1-4C) hydroxy alkyl, miscellaneous Ar³Or phenyl；Wherein (1-4C) examples of alkyl such as methyl； (1-4C) hydroxy alkyl example includes CH₂CH₂OH or CH₂CH₂CH₂OH；Miscellaneous Ar³It is 5 to 6 yuan of hetero-aromatic rings, it is independent with 1-2 Ring hetero atom selected from the N and O and one or more substituent groups for being optionally independently selected from (1-4C) alkyl replace, specific example Including isozole ring, dimethyl isoxazole base etc.；Wherein the phenyl is optionally independently selected from halogen, CF₃, one of CN or Multiple substituent groups replace；

Miscellaneous Ar²It is 5 to 6 yuan of hetero-aromatic rings, is independently selected from N's and S wherein having at least one theheterocyclic nitrogen atom and optionally having Second ring hetero atom, wherein the hetero-aromatic ring is optionally independently selected from one or more substituent groups of (1-4C) alkyl, halogen Replace；

X is selected from

Y is selected from O, NH or NR⁴；

R⁴Selected from (1-6C) alkyl.

In some embodiments of the present invention, R¹Selected from phenyl, it is optionally independently selected from halogen, CF₃、CHF₂'s One or more substituent groups replace.Further, R¹Selected from the phenyl replaced by one or two fluorine atom or by a fluorine original Son and CF₃Substituted phenyl.Specific example, R¹It is to difluorophenyl.

In some embodiments of the present invention, R²Selected from NR^aR^b, it is preferable that the NR^aR^bMiddle R^aSelected from H, R^bSelected from benzene Base, phenyl is optionally by halogen, CF₃One or more substituent groups replace or the NR^aR^bBeing formed has the 4 of theheterocyclic nitrogen atom Circle heterocyclic ring replaces: halogen, OH, (1- wherein the optional ground of the heterocycle is independently selected from a kind of or multiple substituent groups below 4C) alkyl, (1-4C) alkoxy or NR^aR^b5 to 6 circle heterocyclic rings are formed, the heterocycle with theheterocyclic nitrogen atom is formed, the heterocycle is appointed Choosing is independently selected from one or more substituent groups below and is replaced: OH, halogen, CF₃, (1-4C) alkyl.

In a particular embodiment, by NR^aR^bThe group of expression includes with flowering structure:

In view of further increasing inhibitory activity or druggability, NR^aR^bIt is preferably selected from

In some embodiments of the present invention, R²It is (1-4C) hydroxy alkyl.Specific example includes C (CH₃)₂OH and C (CH₃)₂CH₂OH。

In some embodiments of the present invention, R²It is-(1-4C alkyl) Ar¹.Wherein miscellaneous Ar¹It is that there is 1 to 3 ring nitrogen 5 yuan of hetero-aromatic rings of atom.Miscellaneous Ar¹Example be triazole ring, such as 1,2,4- triazole rings.The example of (1-4C) moieties includes Methylene, ethylidene, dimethylmethylene and similar group.

In some embodiments of the present invention, R²It is miscellaneous Ar².The miscellaneous Ar²It is 6 yuan of hetero-aromatic rings, wherein there is at least one Theheterocyclic nitrogen atom and optionally has and be independently selected from the second ring hetero atom of N and S.Miscellaneous Ar²Example include pyridyl ring, pyrimidine radicals Ring, pyrazine basic ring, pyrazoles basic ring, imidazoles basic ring and thiazole basic ring.By miscellaneous Ar²The R of expression²Specific structure include with flowering structure:

Preferably, by miscellaneous Ar²The R of expression²For

In some embodiments of the present invention, R²It is miscellaneous Cyc¹.Miscellaneous Cyc¹Example include carbon connection azetidine Base, pyrrolidinyl and piperidines basic ring are optionally independently selected from (1-4C alkyl), CO₂The one or more of (1-4C alkyl) Substituent group replaces.The example of substituent group includes methyl, ethyl, propyl, CO₂Me、CO₂Et and CO₂C(CH₃)₃.In a kind of embodiment party In formula, miscellaneous Cyc¹Optionally replaced by one or two of described substituent group.

In some embodiments of the present invention, R²It is miscellaneous Cyc².Miscellaneous Cyc²Example include optionally be selected from (1-4C) The pyridone ring or pyridazinone ring that the substituent group of alkyl (such as methyl or ethyl) replaces.

In some embodiments of the present invention, Y is selected from O, NH or NCH₃、NCH₂CH₃.It is steady in order to improve compound metabolism It is qualitative and active, it is preferable that Y NH.

In some embodiments of the present invention, X is selected from Preferably, for pharmacodynamics performance is further increased, X is selected fromPhase For, when X is selected fromWhen, the IC50 value of compound is bigger than normal.

In some embodiments of the present invention, compound has the following structure and spatial configuration:

In order to obtain higher pharmacodynamics performance and metabolic stability performance, In some embodiments of the present invention, preferably Ground, compound has the following structure and spatial configuration:

The compound of logical formula (I) includes its pharmaceutically acceptable salt.Pharmaceutically acceptable salt of the present invention is Inorganic salts or organic salt, inorganic salts include hydrochloride, hydrobromate, hydriodate, sulfate, disulfate, nitrate, phosphoric acid Salt, acid phosphate；The organic salt is selected from acetate, trifluoroacetate, propionate, acetonate, oxyacetate, ethanedioic acid Salt, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, mesylate, with sulphur Hydrochlorate, benzene sulfonate, salicylate.

Further, the compound of Formulas I or their salt can be separated in the form of solvate, and therefore any this molten Agent closes object and belongs to the scope of the present invention.

The compound of logical formula (I) further includes chemical combination only different in terms of the presence of one or more isotope enrichment atoms Object.For example, the compound of the present invention includes wherein one or more hydrogen atoms by deuterium or tritium substitution or one or more carbon atoms The compound replaced by the carbon that 13C- or 14C- is enriched with, within the scope of the invention.

The present invention also provides the preparation methods of compound described in logical formula (I) comprising:

It is for wherein XLogical formula (I) compound, make the respective compound of Formula II with the change of formula III Conjunction object reacts in the presence of coupling agent and/or alkali obtains the compound of formula IV a；

Wherein R⁵It is independently halogen, hydroxyl, methoxyl group, ethyoxyl, propoxyl group, OTs, OMs etc..

Formula IV a and formula HNR^aR^bCompound reaction preparation wherein X beLogical formula (I) compound.

If desired, any blocking group is added or removes, and if desired, forming salt or solvated compounds.

It is for wherein XLogical formula (I) compound, make Formula II respective compound generate isothiocyanates after With with formula HNR^aR^bCompound reaction, or after the compound of Formula II is reacted with thiophosgene again with HNR^aR^bCompound it is anti- It answers, or obtains X under the action of other condensing agents and beLogical formula (I) compound.

It is for wherein XFormulas I compound, make the respective compound and HNR of Formula II^aR^bCompound generate The R replaced again with amino after urea or thiocarbamide³Reaction obtains X and isLogical formula (I) compound.

It is for wherein XFormulas I compound, make Formula II respective compound and formula HNR^aR^bChange Object is closed under the action of sulphinyl chlorine or sulfonic acid chloride, obtaining X isLogical formula (I) compound.

It is for wherein XFormulas I compound, make the respective compound of Formula II with the chemical combination of Formula IV Under the action of coupling agent and/or alkali, obtain X is objectLogical formula (I) compound.

R⁶It is independently halogen, hydroxyl, OTs, OMs etc..

The coupling agent referred in the present invention includes 2- (7- aoxidizes benzotriazole)-N, N, N', N'- tetramethylurea hexafluoro Phosphate (HATU), benzotriazole-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester (HBTU), O- benzotriazole-N, N, N', N'- tetramethylurea tetrafluoro boric acid (TBTU), N, N- dicyclohexylcarbodiimide (DCC), 1- (3- dimethylaminopropyl)- 3- ethyl carbodiimide (DIEC) and any other amide coupling agents well-known to those skilled in the art.Suitable alkali includes uncle Amine base such as diisopropylethylamine (DIEA) and triethylamine, inorganic base etc..

The present invention also provides pharmaceutical compositions, lead to formula (I) compound comprising aforementioned present invention or its is pharmaceutically acceptable Salt and pharmaceutically acceptable carrier, excipient or diluent.

The compounds of this invention or its pharmaceutically acceptable salt may be formulated for the solid pharmaceutical preparation of oral administration, packet Include, but be not limited to capsule, tablet, pill, powder, granule etc..In these solid dosage forms, the present invention leads to formula (I) chemical combination Object is mixed as active constituent at least one conventional inert excipients (or carrier), for example, with sodium citrate or Dicalcium Phosphate. Or it is mixed with subordinate's ingredient: (1) filler or solubilizer, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid etc.； (2) adhesive, for example, hydroxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, Arabic gum etc.；(3) Moisturizer, for example, glycerol etc.；(4) disintegrating agent, such as agar, calcium carbonate, potato starch or tapioca, alginic acid, Mou Xiefu Close silicate and sodium carbonate etc.；(5) retarding solvent, such as paraffin etc.；(6) absorbsion accelerator, such as quaternary ammonium compound etc.；(7) moisten Humectant, such as cetanol and glycerin monostearate etc.；(8) adsorbent, for example, kaolin etc.；(9) lubricant, for example, sliding Or mixtures thereof stone, calcium stearate, solid polyethylene glycol, lauryl sodium sulfate etc.,.Capsule, tablet can also wrap in pill Containing buffer.

Coating and shell material such as enteric can be used in the solid dosage forms such as tablet, sugar-pill, capsule, pill and granule Clothing and other coatings of material crystal form or microencapsulation well known in the art.They may include opacifying agent, also, in this composition The release of active constituent can discharge in certain gastral a part in a delayed fashion.It is adoptable embedding component example be Polymeric material and wax material.When necessary, active constituent can also form microcapsules with one or more of above-mentioned excipient Form.

The compounds of this invention or its pharmaceutically acceptable salt may be formulated for the liquid dosage form of oral administration, packet Include, but be not limited to pharmaceutically acceptable lotion, solution, suspension, syrup, tincture etc..In addition to the general formula as active constituent (I) outside compound or its pharmaceutically acceptable salt, liquid dosage form may include the inert diluent routinely used in this field, example Such as water and other solvents, solubilizer and emulsifier, for example, ethyl alcohol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3- Butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn oil, olive oil, castor oil, sesame oil etc. Or mixture of these substances etc..Other than these inert diluents, fluid present invention dosage form may also comprise conven-tional adjuvants, such as moisten Humectant, emulsifier and suspending agent, sweetener, corrigent and fragrance etc..

The suspending agent includes, for example, ethoxylated octadecanol, polyoxyethylene sorbitol and anhydro sorbitol, crystallite The mixture of cellulose, agar etc. or these substances.

The compounds of this invention and its pharmaceutically acceptable salt are configurable to the dosage form for parental injection, including, But it is not limited to physiologically acceptable sterile, aqueous or anhydrous solution, dispersion liquid, suspension or lotion, and for re-dissolving At the aseptic powdery of sterile Injectable solution and dispersion liquid.Suitable carrier, diluent, solvent, excipient include water, second Alcohol, polyalcohol and its suitable mixture.

The compounds of this invention or its pharmaceutically acceptable salt are configurable to the dosage form for local administration, including such as soft Paste, powder, suppository, drops, stock solution and inhalant etc..The present invention as active constituent leads to formula (I) compound or its medicine On acceptable salt aseptically with physiologically acceptable carrier and optional preservative, buffer, and when necessary The propellant that may be needed is mixed together.

Pharmaceutical composition of the invention includes logical formula (I) compound or its pharmaceutically acceptable salt as active constituent, And pharmaceutically acceptable carrier, excipient, diluent.When preparing pharmaceutical composition, usually by the logical formula (I) of the present invention It closes object or its pharmaceutically acceptable salt is mixed with pharmaceutically acceptable carrier, excipient or diluent.Its formula of (I) chemical combination The content of object or its pharmaceutically acceptable salt can be 0.01-1000mg, such as 0.05-800mg, 0.1-500mg, 0.01- 300mg, 0.01-200mg, 0.05-150mg, 0.05-50mg etc..

The present invention also provides logical formula (I) compound or its pharmaceutically acceptable salt in preparation for treating mammal Cancer, pain, inflammation, neurodegenerative disease, infection by Trypanosoma cruzi or osteolytic disease drug in purposes.

Logical formula (I) compound or its pharmaceutically acceptable salt are used by inhibiting tropomyosin kinases (Trk) activity In cancer, pain, inflammation, neurodegenerative disease, infection by Trypanosoma cruzi or osteolytic disease that treatment mammal includes people Method, the method includes to the mammal include people apply therapeutically effective amount above-mentioned Formulas I compound or its pharmacy Upper acceptable salt.

" therapeutically effective amount " is effectively to generate biology or medicine response in individual (such as to reduce or inhibit enzyme activity albumen living Property, either improve symptom, mitigate the patient's condition, slow down or postpone progression of disease or prevention disease) the compound of the present invention amount.

Cancer mentioned by the present invention, including melanoma, non-small cell lung cancer, thyroid cancer, acute myeloid leukemia, Glioblastoma, astrocytoma and medulloblastoma, colon cancer, neuroblastoma, oophoroma, breast cancer, prostate cancer, Cancer of pancreas, Huppert's disease and maxicell neuroendocrine tumors.

Pain mentioned by the present invention is related to cancer, surgical operation, fracture, the ostalgia as caused by metastases, Bones and joints Inflammation, psoriatic arthritis, rheumatoid arthritis, interstitial cystitis, chronic pancreatitis, splanchnodynia, inflammatory pain, migraine, Chronic low back pain, painful bladder syndrome and neuropathic pain.

Inflammation mentioned by the present invention includes asthma, interstitial cystitis, ulcerative colitis, Crohn disease, atopy Dermatitis, eczema and chronic eczema.

It includes people that the compound of the present invention or its pharmaceutically acceptable salt, which can deliver medicine to mammal, can be taken orally, directly Administration in intestines, parenteral (intravenous, intramuscular or subcutaneous), local administration (pulvis, ointment, drops) or tumor.

Compound of the present invention or its pharmaceutically acceptable salt can be administered alone, or pharmaceutically may be used with other The therapeutic agent of receiving is administered, with other antitumor combinations.This combination therapy can by simultaneously, sequence or be used separately The each component for the treatment of is realized.The therapeutic agent includes but is not limited to: acting on the anti-tumor drug of DNA chemical structure, such as Cis-platinum influences the anti-tumor drug such as methotrexate (MTX), 5 FU 5 fluorouracil etc. of nucleotide synthesis, influences the antineoplastic of transcribed nucleic acid Object such as adriamycin, Epi-ADM, aclacinomycin etc. acts on the anti-tumor drug such as taxol, Changchun of tubulin synthesis Rui Bin etc., arimedex such as aminoglutethimide, Letrozole, auspicious Ningde etc., cell-signaling pathways inhibitor such as epidermal growth Factor receptor inhibitor Imatinib (Imatinib), Gefitinib (Gefitinib), Tarceva (Erlotinib) etc..It is anti- Tumour monoclonal antibody, immunosuppressor PD-1, PD-L1 etc., each ingredient to be combined can simultaneously or sequentially be given, with unitary system dosage form Formula is given in the form of different preparations.The combination not only includes one kind of the compounds of this invention or the group of other activating agents It closes, and the combination of other activating agents of two or more including the compounds of this invention.

The one or more other drugs acted on other same or different role mechanisms play are applied in combination.This, which combines, controls Treat can by simultaneously, sequence or be used separately each component for the treatment of and realize.The therapeutic agent includes but is not limited to: steroid Dexamethasone, cortisone, fluticasone；Antalgesic, aspirin, brufen, Indomethacin and opioids.

Embodiment

The following example illustrates rather than limit the synthetic method of logical formula (I) compound.Temperature is degree Celsius.If In addition do not illustrate, all evaporations carry out under reduced pressure.If not stated otherwise, otherwise reagent is from commercial suppliers It buys and is used without being further purified.The structure of final product, intermediate and raw material by standard method of analysis confirm, such as Elemental analysis, Spectral Characteristics Analysis, such as MS, NMR.The abbreviation used is this field Conventional abbreviations.Intermediate A

(R) -5- (2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-yl) amido) -4- first The preparation of oxygroup ring butyl- 3- alkene -1,2- diketone

Methanol (50mL) is added in the flask of 100mL, adds (R) -5- (2- (2,5- difluorophenyl) pyrrolidines -1- Base) pyrrolo- [1,5-a] pyrimidine -3- amine (6.4g, 20.5mmol), add squaric acid (2.32g, 16.3mmol), is heated to reflux 6-8h, gradually there is solid precipitation, and cooled and filtered obtains yellow solid 2.5g (yield 28.6%).Structural formula is as follows:

Intermediate B

(R) -5- (2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-yl) amido) -4- second The preparation of oxygroup ring butyl- 3- alkene -1,2- diketone

Ethyl alcohol (50mL) is added in the flask of 100mL, adds (R) -5- (2- (2,5- difluorophenyl) pyrrolidines -1- Base) pyrrolo- [1,5-a] pyrimidine -3- amine (6.4g, 20.5mmol), add squaric acid (2.32g, 13.6mmol), is heated to reflux 6-8h, gradually there is solid precipitation, and cooled and filtered obtains yellow solid 2.7g (yield 30.9%).Structural formula is as follows:

Intermediate C

(R) -1- ((5- (2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrrolo- [1,5-a] pyrimidin-3-yl) amido) ammonia Formoxyl) cyclopropyl-carboxylic acid preparation

1,1- cyclobutyl dioctyl phthalate (10.0g, 69.4mmol) is added in anhydrous THF (100mL).Under nitrogen protection by Be added dropwise to triethylamine (7.02g, 69.4mmol 1.0eq) and stir 30 minutes at 0 DEG C, be then added thionyl chloride (8.25g, 69.4mmol, 1.0eq) it is stirred 30 minutes at 0 DEG C again.(R) -5- (2- is added dropwise into reaction solution again under nitrogen protection (2,5- difluorophenyl) pyrrolidin-1-yl) pyrrolo- [1,5-a] pyrimidine -3- amine (21.7g, 76.3mmol) anhydrous THF (50mL) solution, stirs 10h at 10-15 DEG C.Reaction solution is diluted with ethyl acetate, and extracts (pH > 10) with the NaOH of 2N.Water The HCl that 2N is mutually added dropwise adjusts pH to 2-3, is then extracted with ethyl acetate.Organic phase is dry to hang and dry obtains 23.0g off-white color Solid (yield 71%).Structural formula is as follows:

Intermediate D

(R) system of -5- (2- (2,5- difluorophenyl) pyrrolidin-1-yl) -3- isothiocyanic acid base pyrazolo [1,5-a] pyrimidine It is standby

Thiocarburyl chloride (0.76g, 10mmol) is added in methylene chloride (8mL) solution in three-necked flask, is cooled down To 0 DEG C, by (R) -5- (2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidine -3- amine (2.83g, Methylene chloride (10mL) 9mmol) is added thereto.Sodium hydroxide (1.2g, 30mmol) is added in reaction solution.React liquid chamber Temperature stirring 3h, reaction solution washing, dry, evaporating solvent under reduced pressure obtains 2.1g product (yield 63%).Structural formula is as follows:

Embodiment 1

3- ((5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrrolo- [1,5-a] pyrimidin-3-yl) amido) - 4- ((S) -3- hydroxyl pyrrolidine -1- base) ring butyl- 3- alkene -1,2- diketone

It is (R) -5- (2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-yl by intermediate A) Amido) intermediate B of -4- methoxy basic ring butyl- 3- alkene -1,2- diketone (0.5g, 1.18mmol) or equivalent is dissolved in methanol In (10mL), it is added (S) -3- hydroxy-pyrrolidine hydrochloride (0.2g, 2.36mmol), under microwave condition in closed container, 1.5h is reacted at 80 DEG C, is cooled to room temperature, there is solid generation, is filtered, a small amount of methylene chloride is added after filtrate is outstanding dry, standing has solid Body is precipitated, and 0.12g product (yield 21%) is obtained by filtration.¹H-NMR (400MHz, CDCl₃) δ: 6.31 (s, 1H), 8.17 (d, 2H), 7.0 (m, 1H), 6.91 (m, 1H), 6.76 (m, 1H), 6.22 (s, 1H), 5.97 (s, 1H), 5.30 (s, 1H), 4.56 (d, 1H),3.91-3.53(m,5H),2.5(m,1H),2.13-2.08(m,6H),MS m/z 481.2(M+1)⁺.Structural formula is as follows:

Embodiment 1A

3- ((5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-yl) amido) - 4- ((S) -3- hydroxyl pyrrolidine -1- base) ring butyl- 3- alkene -1,2- diketone sulfate

At room temperature, to 3- ((5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidine - 3- yl) amido) -4- ((S) -3- hydroxyl pyrrolidine -1- base) ring butyl- 3- alkene -1,2- diketone (0.48g, 1.0mmol) methanol Sulfuric acid (5mL, 1mmol) in methyl alcohol is added in (20mL) solution.Acquired solution stirs 1 hour, is then concentrated to get yellow Solid 3- ((5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrrolo- [1,5-a] pyrimidin-3-yl) amido) -4- ((S) -3- hydroxyl pyrrolidine -1- base) ring butyl- 3- alkene -1,2- diketone sulfate 0.42g (yield 72%).Structural formula is as follows:

Embodiment 1B

3- ((5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrrolo- [1,5-a] pyrimidin-3-yl) amido) - 4- ((S) -3- hydroxyl pyrrolidine -1- base) ring butyl- 3- alkene -1,2- dione hydrochloride

At room temperature, to 3- ((5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidine - 3- yl) amido) -4- ((S) -3- hydroxyl pyrrolidine -1- base) ring butyl- 3- alkene -1,2- diketone (0.48g, 1.0mmol) methanol It is added in (20mL) solution at hydrogen chloride methanol solution (5mL, 1mmol).Acquired solution stirs 1 hour, is then concentrated to get Huang Color solid 3- ((5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrrolo- [1,5-a] pyrimidin-3-yl) amido) -4- ((S) -3- hydroxyl pyrrolidine -1- base) ring butyl- 3- alkene -1,2- dione hydrochloride 0.44g (yield 86%).

Embodiment 2

By (R) -5- (2- (the fluoro- 2- trifluorophenyl of 5-) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-yl) amido) - 4- ethoxy basic ring butyl- 3- alkene -1,2- diketone (commercially available, 0.36g, 1.0mmol) is dissolved in methanol (10mL), and (S) -3- hydroxyl is added Base pyrrolidine hydrochloride (0.17g, 2.0mmol) reacts 1.5h at 80 DEG C, is cooled under microwave condition in closed container Room temperature has solid generation, filtering, and a small amount of methylene chloride is added after filtrate is outstanding dry, and standing has solid precipitation, 0.10g is obtained by filtration Product (yield 22%).MS m/z 531.2(M+1)⁺.Structural formula is as follows:

By operation synthetic example 3-9 (referring to table 1) similar with 1 basic operation of embodiment, to obtain desired production Object.

The structure and mass spectrometric data of 1 embodiment 3-8 of table

Embodiment 9

N- (5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-yl) -1- ((S) - 3- hydroxyl pyrrolidine -1- carboxyl) cyclopropyl carboxamide

It is (R) -1- ((5- (2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrrolo- [1,5-a] pyrimidine-by intermediate C 3- yl) amido) carbamyl) cyclopropyl-carboxylic acid (21.4g, 50mmol) is dissolved in 150mL THF, instill 0.2mL DMF, ice (COCl) is instilled in water-bath₂(6.3g, 50mmol) is added dropwise process temperature and is no more than 15 DEG C, after dripping off, and 15 DEG C are stirred 4 hours, are turned It is stand-by to enter dropping funel.(S) -3- hydroxy-pyrrolidine hydrochloride (4.9g, 40mmol), potassium carbonate (27.6g, 200mmol) is successively It is added in the mixed solution of 150mL water and 300mL THF, instills the above acyl chlorides, this process temperature is no more than 25 DEG C, after dripping off It is stirred at room temperature 2h hours, is added water (500mL), is stirred at room temperature 8 hours, solid is precipitated, filter, obtain off-white powder, directly add Enter into the mixed solution of 500mL THF and 500mL water, flow back 1 hour, be cooled to room temperature, solid is precipitated, filters, dry White solid 14.3g (yield 58%).

¹H-NMR (400MHz, CDCl₃) δ: 6.33 (s, 1H), 8.17 (d, 2H), 7.16 (m, 1H), 6.92 (m, 1H), 6.70 (m, 1H), 6.23 (s, 1H), 5.97 (s, 1H), 5.28 (s, 1H), 4.57 (d, 1H), 3.91-3.53 (m, 5H), 2.5 (m, 1H), 2.12-2.08(m,6H),1.49-1.47(m,4H).MS m/z 497.2(M+1)⁺.Structural formula is as follows:

Embodiment 9A

N- (5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-yl) -1- ((S) - 3- hydroxyl pyrrolidine -1- carboxyl) cyclopropyl carboxamide sulfate

At room temperature, to 3- ((5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrrolo- [1,5-a] pyrimidine - 3- yl) amido) -4- ((S) -3- hydroxyl pyrrolidine -1- base) ring butyl- 3- alkene -1,2- diketone (0.50g, 1.0mmol) methanol Sulfuric acid (5mL, 1mmol) in methyl alcohol is added in (20mL) solution.Acquired solution stirs 1 hour, is then concentrated to get yellow Solid 3- ((5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrrolo- [1,5-a] pyrimidin-3-yl) amido) -4- ((S) -3- hydroxyl pyrrolidine -1- base) ring butyl- 3- alkene -1,2- diketone sulfate 0.42g (yield 72%).Structural formula is as follows:

Embodiment 9B

N- (5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-yl) -1- ((S) - 3- hydroxyl pyrrolidine -1- carboxyl) cyclopropyl carboxamide hydrochloride

At room temperature, to N- (5- ((R) -2- (2,5- difluorophenyl) pyrrolo- [1,5-a] pyrimidin-3-yl) -1- ((S) - 3- hydroxyl pyrrolidine -1- carboxyl) cyclopropyl carboxamide (0.50g, 1.0mmol) methanol (20mL) solution in be added in hydrogen chloride Methanol solution (5mL, 1mmol).Acquired solution stirs 1 hour, is then concentrated to get yellow solid 3- ((5- ((R) -2- (2,5- Difluorophenyl) pyrrolidin-1-yl) pyrrolo- [1,5-a] pyrimidin-3-yl) amido) -4- ((S) -3- hydroxyl pyrrolidine -1- base) ring Butyl- 3- alkene -1,2- dione hydrochloride 0.45g (yield 83%).Structural formula is as follows:

Embodiment 9C

N- (5- ((R) -2- (2,5- difluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-yl) -1- ((S) - 3- hydroxyl pyrrolidine -1- carboxyl) cyclopropyl carboxamide L MALIC ACID salt

N- (5- ((R) -2- (2,5- difluorophenyl) pyrrolo- [1,5-a] pyrimidin-3-yl) -1- ((S) -3- hydroxypyrrole Alkane -1- carboxyl) cyclopropyl carboxamide (0.50g, 1.0mmol) is dissolved in 10mL butanone；L MALIC ACID (0.16g, 1.2mmol) is dissolved in 2mL water, aqueous solution of malic acid are slowly added into butanone solution, are warming up to 50-55 DEG C of interior temperature and are stirred 2 hours, return stirring 4-5 Hour.It is cooled to room temperature, stirring for 24 hours, filters, and filter cake is washed with 2mL, and the washing of 2mL butanone is dried in vacuo to obtain white solid 0.5g (yield 80%).Structural formula is as follows:

Embodiment 10

(S)-N- (5- ((R) -2- (2,5- difluorophenyl) pyrroles -1- base) pyrazolo [1,5-a] pyrimidin-3-yl) -3- hydroxyl Base pyrroles's -1- thiocarbamide

At 0 DEG C, to the DMF for using the washed sodium hydride of n-hexane (60% oil dispersion, 240mg, 6.0mmol) in advance It is (R) -5- (2- (2,5- difluorophenyl) pyrrolidin-1-yl) -3- isothiocyanic acid base pyrazolo that intermediate D is added in (10mL) DMF (5mL) solution of [1,5-a] pyrimidine (3mmol).At 0 DEG C, after stirring 15min under argon gas, (S) -3- hydroxyl is added dropwise at 0 DEG C Simultaneously 1h is stirred at room temperature in the DMF solution (5mL, 3mmol) of base pyrrolidine hydrochloride.0.5M sodium dihydrogen phosphate is added to terminate instead It answers.Mixture is extracted with ethyl acetate, and is washed with water and dry with sodium sulphate, crude product obtains purpose compound with recrystallizing methanol 0.99g (yield 75%), MS m/z 445.2 (M+1)⁺.Structural formula is as follows:

By operation synthetic example 11-14 (referring to table 2) similar with 9 basic operation of embodiment, desired product is obtained.

The structure and mass spectrometric data of 2 embodiment 11-14 of table

Intermediate E

At 0 DEG C, argon gas, under stirring condition, to embodiment 13 (1.0g, 2.3mmol) and triethylamine (1mL, 6.9mmol) Methane sulfonyl chloride (4.6mmol) is added dropwise in dichloromethane solution.15min is stirred at 0 DEG C.Detection has been reacted.Mixture passes through Column chromatographic purifying (dichloromethane eluent) obtains target compound 0.7g (yield 73%), MS m/z 417.2 (M+1)⁺.Structure Formula is as follows:

Embodiment 15

Under stirring, in being added dropwise in the acetonitrile mixture of cyanamide (560mg, 13.33mmol) and Huinig alkali (2.6mL) Acetonitrile (30mL) solution of mesosome E (0.7g, 1.69mmol).Reaction 15min is stirred at room temperature, solvent is then removed under reduced pressure, Residue is hydrolyzed with 0.5M sodium dihydrogen phosphate.The acetic acid ethyl acetate extract of water solution mixture 0.5M sodium dihydrogen phosphate and salt water Washing filters after dry and distills to obtain crude product, and column chromatographs to obtain target compound 0.46g (yield 61%), 445.2 (M+ of MS m/z 1)+.Structural formula is as follows:

By operation synthetic example 16-18 (referring to table 3) similar with 15 basic operation of embodiment, desired production is obtained Object.

The structure and mass spectrometric data of 3 embodiment 16-18 of table

Embodiment 19

TrkA ELISA measurement

TrkA kinase activity is evaluated using enzyme linked immunosorbent assay (ELISA) (ELISA) in the presence of the inhibitors.It uses 0.025mg/mL poly- (Glu, Ala, Tyr；6:3:1；Sigma P3899) solution coating Immulon4HBX384- hole microtitration Plate (Thermo).At ambient temperature by the test compound of each concentration, 2.5nM TrkA in the plate of coating (Invitrogen, the recombined human Trk of histidine mark, cytoplasmic domains) and 500 μm of ol ATP are incubated for 25 minutes, are shaken simultaneously It swings.Measurement buffer is made of 15mM MOPS pH7.5,0.005% (v/v) Triton X-100 and 5mM MgCl2.Pass through use PBS washing containing 0.1% (v/v) tween (Tween) 20, removes reaction mixture from plate.Use coupling horseradish peroxidase 0.2 μ g/mL phosphotyrosine monoclonal antibody specific (clone PY20) combine TMB peroxidase material system (KPL) Detect the anti-product of phosphorylation.After adding 1M phosphoric acid, via the absorptance quantitative chromogenic material color intensity under 450nm.Use 4 Or 5- parameter logistic (logistic) curve matching calculates IC50 value.

In the measurement, chemical combination of the invention has the average IC lower than 1000nM₅₀.Certain compounds, which have, to be lower than The average IC of 100nM₅₀.Table 4 provides the specific IC that the compound of the present invention is tested in the measurement₅₀Value.

The specific IC of 4 the compound of the present invention of table₅₀Value

Embodiment number	TrkA ELISA enzyme IC₅₀(nM)
		1	3.7
2	6.5
		3	10.3
4	87.5
		5	3.7
6	2.0
		7	28.4
8	47.1
		9	97.8
10	117.6
		11	239.4
12	317.6
		13	129.5
14	137.2
		15	23.1
16	36.8
		17	181.5
18	233.1

Embodiment 20

The evaluation of compound stability is carried out using human body liver microsomes.

The hepatomicrosome enzyme stability of embodiment compound and Larotrectinib (LOXO-101) are compared.

Measurement system: the metabolic stability of the compounds of this invention utilizes the liver microsomes 1mM mixed by men and women NADPH is tested.Sample is analyzed using mass spectrograph.HRMS is used to determine that peak area response ratio (to correspond to test The peak area of compound or reference material is divided by target peak area in analyzing) without operation standard curve.It is all in order to detect Possible metabolin carries out HRMS scanning within the scope of m/z appropriate.

Determination condition: the measurement is carried out with primary be incubated for (N=1).Test compound is being contained into 0.5 milli at 37 DEG C It is incubated in the buffer of grams per milliliter liver microsomal protein.By the way that co-factor initiation reaction is added, and in 0,2,4,8,16,24, 36, it samples within 48 hours, be incubated for positive control (5 μM of cortisols) in parallel and sampled in 0,2,4,8,16,24,36,48 hour.

Quality measurement control: parallel to carry out control compound cortisol to confirm the enzymatic activity of (liver) microsome.Finally After time point, confirm that NADPH is added in reaction mixture using fluorimetry.The T1/2 of reference material meets acceptable Internal standard.

Analysis method:

Liquid-phase chromatographic column: Thermo BDS Hypersil C18 30X2.0mm, has guard column M.P., buffering by 3 μm Liquid: 25mM formic acid meets buffer, pH 3.5；

Water phase (A): 90% water, 10% buffer；

Organic phase (B): 90% acetonitrile, 10% buffer；

Flow velocity: 300 mul/min

Autosampler: 10 microlitres of volume injected

Gradient program is referring to table 5.

5 Gradient program of table

Time (minute)	%A	%B
			0.0	100	0
1.5	0	100
			2.0	0	100
2.1	100	0
			3.5	100	0

By using human liver microsome body, embodiment 1,2,4,5,6,7,8,16 shows to be greater than 48 as described in the present invention The metabolic half life of hour, embodiment 3,9,17,18 show the metabolic half life between 36-48 hours, noticeably greater than Larotrectinib (LOXO-101) 23 hours metabolic half life, the metabolic half life of embodiment 15 is between 24-36 hour Between, the metabolic half life of slightly above Larotrectinib (LOXO-101).The results show that relatively long metabolic half life makes Obtaining them has the potential for reducing treatment doses and expanding administration time interval.

By the specific IC of the compound of embodiment 1-18₅₀Value and metabolic half life data it is found that for lead to formula (I) class chemical combination For object, linking group and substituent group such as X group, R²The selection of group and R¹In substituent group and its position for change The pharmacodynamics performance and metabolic stability for closing object have important influence.

Although the specific embodiment before the present invention passes through illustrates, it should not be constructed as being so limited；But this hair It is bright to cover disclosed general aspect before.A variety of modifications can be carried out under without departing substantially from the spirit and scope of the present invention and are had a variety of Embodiment.

Claims

1. a kind of compound or its pharmaceutically acceptable salt with logical formula (I),

Wherein:

R¹It is (i) phenyl, is optionally independently selected from halogen, (1-4C) alkoxy, CF₃、CHF₂, one or more replace Base replaces；Or (ii) 5 to 6 yuan of heteroaromatics, there is the heteroatom selected from N and S, wherein the heteroaromatic is optionally by one Or multiple halogen atoms replace；

R²Selected from NR^aR^b, (1-4C) alkyl, (1-4C) fluoroalkyl, (1-4C) hydroxy alkyl ,-(1-4C alkyl) miscellaneous Ar¹, miscellaneous Ar²、 Miscellaneous Cyc¹, miscellaneous Cyc²Or it is optional by F, CF₃、OH、NH₂, OMe, OEt, NHMe replace (3-6C) naphthenic base；

R^aIt is H or (1-6C) alkyl；

R^bIt is H, (1-4C) alkyl, (1-4C) hydroxy alkyl, miscellaneous Ar³Or phenyl, wherein the phenyl is optionally independently selected from halogen Element, CF₃, CN one or more substituent groups replace；

Or NR^aR^bFormed have theheterocyclic nitrogen atom 4 circle heterocyclic rings, wherein the heterocycle be optionally independently selected from one below or Multiple substituent groups replace: halogen, OH, (1-4C) alkyl, (1-4C) alkoxy；

Or NR^aR^b5 to 6 circle heterocyclic rings are formed, the heterocycle is with theheterocyclic nitrogen atom and optionally with selected from N, O and SO₂Second The heterocycle that ring hetero atom or group are formed takes wherein the heterocycle is optionally independently selected from one or more substituent groups below Generation: OH, halogen, CF₃, (1-4C) alkyl, NH₂,COOH；

Miscellaneous Ar²It is 5 to 6 yuan of hetero-aromatic rings, wherein having at least one theheterocyclic nitrogen atom and optionally there is the second ring for being independently selected from N and S Hetero atom, wherein the hetero-aromatic ring is optionally independently selected from (1-4C) alkyl, one or more substituent groups of halogen replace；

Miscellaneous Cyc¹It is 4 to 6 yuan of azacyclo-s of carbon connection, is optionally independently selected from (1-4C) alkyl, CO₂The one of (1-4C alkyl) A or multiple substituent groups replace；

Miscellaneous Ar³It is 5 to 6 yuan of hetero-aromatic rings, is independently selected from the ring hetero atom of N and O with 1-2 and is optionally independently selected from (1- 4C) one or more substituent groups of alkyl replace；

X is selected from

Y is selected from O, NH or NR⁴；

R⁴Selected from (1-6C) alkyl.

2. compound as described in claim 1 or its pharmaceutically acceptable salt, which is characterized in that R²Selected from NR^aR^b、(1- 4C) alkyl, (1-4C) fluoroalkyl, miscellaneous Ar², miscellaneous Cyc¹, miscellaneous Cyc²。

3. compound as described in claim 1 or its pharmaceutically acceptable salt, which is characterized in that R²Selected from NR^aR^b, miscellaneous Ar², The wherein NR^aR^bMiddle R^aSelected from H, R^bSelected from phenyl, phenyl is optionally by halogen, CF₃One or more substituent groups replace, or NR described in person^aR^b4 circle heterocyclic rings with theheterocyclic nitrogen atom are formed, wherein the optional ground of the heterocycle is independently selected from one kind below Or multiple substituent groups replace: halogen, OH, (1-4C) alkyl, (1-4C) alkoxy or NR^aR^b5 to 6 circle heterocyclic rings are formed, are formed Heterocycle with theheterocyclic nitrogen atom, the heterocycle are optionally independently selected from one or more substituent group below and are replaced: OH, halogen, CF₃, (1-4C) alkyl, wherein the miscellaneous Ar²It is 6 yuan of hetero-aromatic rings, wherein having at least one theheterocyclic nitrogen atom and optionally having independent The second ring hetero atom selected from N and S.

4. compound as claimed in claim 1 or 2 or its pharmaceutically acceptable salt, which is characterized in that R¹Selected from phenyl, Optionally it is independently selected from halogen, CF₃、CHF₂One or more substituent groups replace.

5. compound as claimed in claim 4 or its pharmaceutically acceptable salt, which is characterized in that R¹Selected from by one or two Phenyl that a fluorine atom or chlorine atom replace or by a fluorine atom and CF₃Substituted phenyl.

6. compound as described in claim 1 or its pharmaceutically acceptable salt, which is characterized in that

The compound is selected from:

7. compound as claimed in claim 1 or 2 or its pharmaceutically acceptable salt, wherein described is pharmaceutically acceptable Salt be inorganic salts or organic salt, inorganic salts include hydrochloride, hydrobromate, hydriodate, sulfate, disulfate, nitrate, Phosphate, acid phosphate；The organic salt is selected from acetate, trifluoroacetate, propionate, acetonate, oxyacetate, second Diacid salt, malonate, fumarate, maleate, lactate, malate, citrate, tartrate, mesylate, With sulfonate, benzene sulfonate, salicylate.

8. a kind of pharmaceutical composition, including the described in any item compounds of claim 1-7 or its pharmaceutically acceptable salt with And pharmaceutically acceptable carrier, excipient or diluent.

9. the described in any item compounds of claim 1-7 or its pharmaceutically acceptable salt are in preparation for treating mammal Cancer, pain, inflammation, neurodegenerative disease, infection by Trypanosoma cruzi or osteolytic disease drug in purposes.

10. purposes as claimed in claim 9, wherein the cancer includes melanoma, non-small cell lung cancer, thyroid gland Cancer, acute myeloid leukemia, glioblastoma, astrocytoma and medulloblastoma, colon cancer, neuroblastoma, ovary Cancer, breast cancer, prostate cancer, cancer of pancreas, Huppert's disease and maxicell neuroendocrine tumors；Wherein the pain relates to And cancer, surgical operation, fracture, the ostalgia as caused by metastases, osteoarthritis, psoriatic arthritis, rheumatoid joint Inflammation, interstitial cystitis, chronic pancreatitis, splanchnodynia, inflammatory pain, migraine, chronic low back pain, painful bladder syndrome and mind Through property pain；Wherein the inflammation includes asthma, interstitial cystitis, ulcerative colitis, Crohn disease, atopy skin Scorching, eczema and chronic eczema.