CN109776544A - Pyrazolo [3,4-d] pyrimidines and its preparation method and application - Google Patents

Abstract

In the inhibitor that compound of the present invention is a kind of bruton's tyrosine kinase.In addition, disclosure sets forth the purposes of the drug component comprising the compound and preparation and this kind of kinase inhibitor, can single drug or with other compound drug combinations, for treating by the symptom of kinase mediated or kinases dependence.

Description

Pyrazolo [3,4-d] pyrimidines and its preparation method and application

Technical field

The present invention relates to compound, the method for prepare compound, the pharmaceutical composition and medicament of compound, and by the chemical combination Object for treat, prevent, diagnose the relevant disease of bruton's tyrosine kinase (Bruton ' s tyrosine kinase, Btk), Imbalance or symptom.

Background technique

Protein tyrosine kinase adjusts growth, differentiation, apoptosis of cell etc. one by the signal transduction pathway of control cell Serial physiological and biochemical procedure.Abnormal kinase activity has been directed to many human diseases, including inflammation, autoimmune disease and The diseases such as cancer.(such as gastric cancer, lung cancer and lymthoma) has found the exception of protein tyrosine kinase in general human cancer Expression, protein tyrosine kinase have become one of the important target spot of anti-tumor drug research and development.

Btk is the member of nonreceptor tyrosine kinase Tec family, by PH structural domain, TH structural domain, SH3 structural domain, SH2 Structural domain and 5 part of catalyst structure domain composition.Btk participates in many A signal pathways, rises emphatically to the proliferation of cell, differentiation and apoptosis The regulating and controlling effect wanted, in the signal path of connection cell surface B-cell receptor (B-cell receptor, BCR), with BCR Activation, Btk rely on Syk, Lyn activation, the Btk after activation can cause include the downstream signals such as MAPK, NF κ B activation, it is different The B cell activation or the formation of pathogenic autoantibodies that the signal transduction that normal BCR is mediated can cause mistake to adjust, this will lead to Various autoimmune or diseases associated with inflammation.The sustained activation of Btk is an elder generation of chronic lymphocytic leukemia (CLL) development Certainly condition, unconventionality expression can also promote the survival of activating B cell hypotype in diffusivity large B cell lymphoid tumor (DLBCL).

Btk micromolecular inhibitor can inhibit the proliferation of B lymphoma cell by the activity of inhibition Btk, promote tumour thin The apoptosis of born of the same parents can also inhibit the generation of B cell autoantibody and cell factor, for treatment hematologic malignancies and itself exempt from Epidemic disease dysfunctional disease has good prospect.Have multiple compounds and enter clinical research, for treating B cell lymphoma and white blood Disease etc..The research and development of Pharmacyclics biopharmaceutical company replace Buddhist nun (Ibrutinib) can not as one kind of first listing according to Shandong Inverse Btk micromolecular inhibitor drug, currently used for treatment lymphoma mantle cell, chronic lymphocytic leukemia and macroglobulin Mass formed by blood stasis etc. has apparent curative effect.Other are in the small molecule Btk inhibitor of clinical development, and there are also the ONO- of ONO company 4059, the CC-292 etc. of the ACP-196 and Celgene company of Acerta company.

Currently, will be at according to the highly selective micromolecular inhibitor that Btk kinases is declared publicly for the outstanding clinical effectiveness of Buddhist nun in Shandong For the another hot spot in global new drug development field, also indicate that pyrazolo [3,4-d] miazines structural compounds are that a kind of take orally has The Btk micromolecular inhibitor drug of effect.Therefore, in view for the treatment of tumor disease there is an urgent need to the diversified pyrazoles of research structure And [3,4-d] pyrimidines are of great significance to the new more good Btk micromolecular inhibitor drug of effect is developed.

Summary of the invention

Compound of the present invention, pharmaceutical composition, medicament and method can be used for (a) diagnosis, prevent, treat and Btk Relevant disease, imbalance or symptom；(b) mitigate side effect relevant to Btk or symptom；(c) control relevant to Btk disease, Imbalance or symptom.On the one hand, method set forth herein, compound, pharmaceutical composition and medicament by inhibit Btk inhibitor structure At.

In the first aspect of the present invention, a kind of compound of formula I or its pharmaceutically acceptable salt are provided, solvate, Active metabolite, polymorph, ester, isomers or prodrug:

Wherein:

L₁It is the alkenyl of alkyl, unsubstituted or substituted C2-C6 selected from unsubstituted or substituted C1-C6, unsubstituted or take The alkynyl of the C2-C6 in generation；

L₂Selected from-(CH₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n- NH-, wherein n is 0,1,2,3 or 4；

Ar is selected from unsubstituted or substituted aryl or heteroaryl；

Y is selected from unsubstituted or substituted alkyl radical or 4 yuan, 5 yuan, 6 yuan of cycloalkyl rings；

R₁Selected from H or low alkyl group；

Alternatively, Y and N and R₁It is connected to form a quaternary, five yuan or hexa-member heterocycle；

G is selected from H,

Wherein, R₂、R₃And R₄Separately selected from H, halogen ,-COOH, unsubstituted or substituted low alkyl group, unsubstituted Or the Lower heteroalkyl replaced.

In another preferred example, in which: n is 1 or 2.

In another preferred example, in which: Ar is selected from unsubstituted or substituted aryl；More preferably, it is substituted by meta or para position Replace；More preferably, substituent group is alkoxy or halogen.

In another preferred example, in which: L₁The alkynyl of alkenyl or unsubstituted C2-C6 selected from unsubstituted C2-C6；More Goodly, in which: L₁Selected from vinyl or acetenyl.

In another preferred example, in which: Y and N and R1 are connected to form hexa-member heterocycle.

In another preferred example, in which: G is

In another preferred example, compound provided by the invention is selected from:

In another preferred example, compound provided by the invention is to bruton's tyrosine kinase (Bruton ' s tyrosine Kinase, Btk) there is inhibitory activity.

In the second aspect of the present invention, a kind of pharmaceutical composition is provided, which includes to mention present invention as described above The compound of confession and pharmaceutically acceptable excipient.

In another preferred example, the form of aforementioned pharmaceutical compositions is aqueous dispersion, liquid, Gel miles, syrup, west Agent, medicine slurry, suspension, aerosol, controlled release agent, quick-dissolving agent, effervescent agent, freeze-dried, tablet, powder, pill, sugar-coat be complete, capsule, Sustained release agent, extended release agent, pulse controlled release agent, multiparticulates agent release immediately agent.

In the third aspect of the present invention, a kind of preparation method of compound provided present invention as described above is provided, The method includes the steps: under nitrogen protection, by structure such as I-d compound represented of formula and DMF, palladium catalyst and organic base Mixing, and then reacted at 30-150 DEG C 3-15 hours with structure such as I-c compound represented of formula, obtained structure such as I-e of formula Compound represented is deprotected preparation structure intermediate as shown in I-f of formula in acid condition, then in alkali or condensing agent and In the presence of alkanes solvent, the amide condensed compound for reacting, being provided present invention as described above is carried out with G-Z；

Wherein: L₁、L₂、Ar、Y、R₁, G definition as shown in claim 1；Z is selected from halogen or hydroxyl.

In the fourth aspect of the present invention, provide a kind of compound provided present invention as described above in preparation prevention or Treating inflammation, autoimmune disease (such as rheumatoid arthritis) relevant to aberrant B cell proliferation and/or tumor disease Application in drug.

Accordingly, the present invention provides the more good Btk micromolecular inhibitor drugs of new effect.

Detailed description of the invention

Fig. 1 shows that homogeneous phase time discrimination fluorescence (HTRF) method establishes the kinase activity detection of Btk micromolecular inhibitor Principle.

Fig. 2 shows that homogeneous phase time discrimination fluorescence (HTRF) method establishes the kinase activity detection of Btk micromolecular inhibitor Operating process.

Specific embodiment

Term

If being used for the present patent application without other explanation, including the term in specification and claims, definition is such as Under.It has to be noticed that in the specification and the appended claims, if Wen Zhongwu is clearly dictated otherwise, singular " one It is a " it include plural references.If using mass spectrum, nuclear-magnetism, HPLC, protein chemistry, biochemistry, recombinant DNA skill without other explanation The conventional method of art and pharmacology.In this application, if referring to "and/or" using "or" or "and" without other explanation.

" formula (I) compound " refers to that structural formula is the compound of (I).

" alkyl " refers to aliphatic hydrocarbon hydrocarbon group.The alkyl that moieties can be saturation (refers to without containing any unsaturated single Member, such as carbon-to-carbon double bond or carbon-carbon triple bond) or moieties can be unsaturated alkyl and (refer at least unsaturated single containing one Member).Moieties can be branch or straight chain regardless of saturation or unsaturation.

" alkyl " segment (moiety) can have 1 to 8 carbon atoms (as long as occurring herein, digital scope such as " 1 Refer to 8 " and is providing each integer in range, such as " 1 to 8 carbon atoms ", which refers to, can contain 1 carbon atom, 2 carbon atoms, and 3 A carbon atom etc. until the alkyl comprising 8 carbon atoms, although it is current be defined on no designation number range in the case where, Contain the appearance of term " alkyl ").The alkyl of compound as described herein can be designated as " C₁-C₆Alkyl " is similar It is specified.Illustrate " C₁-C₆Alkyl " refers to one in alkyl chain, two, three, four, five or six carbon atoms.Allusion quotation The alkyl of type includes but is not limited to methyl, ethyl, propyl iso-propyl, butyl isobutyl group, tert-butyl, amyl, hexyl etc..Term " low alkyl group " is similarly used for the group with 1 to 4 carbon atoms.

" alkoxy " group refers to that " alkyl " O- group, alkyl are as defined herein.

" acylamino- " is the chemical segment that molecular formula is-C (=O) NHR or-NHC (=O) R, and R contains alkane selected from group Base, naphthenic base, aryl, heteroaryl (being connected by ring carbon) (are connected) with heterolipid cyclic hydrocarbon by ring carbon.Amide can be amino Acid or the peptide molecule for being connected to formula (I) compound, to form prodrug.Any amino or carbon side in compound as described herein Chain is as required by the amidation of selectivity.See Greene and Wuts, Protective Groups in Organic Synthesis, 3^rdEd., John Wiley&Sons, New York, NY, 1999.

If the term " aryl " used herein refers to aromatic rings, wherein each atom for forming ring is carbon atom.Aryl By five, six, seven, eight, nine or more form ring.Aryl is optionally substituted.On the one hand, aryl be phenyl or Naphthalene.According to structure, aryl can be single free radical or diradical (such as arlydene).On the one hand, aryl is C₆-C₁₀Aryl.

Term " naphthenic base " refers to monocycle or polycyclic aliphatic hydrocarbon, non-aromatic free radical, wherein each atom (such as skeleton of cyclization Atom) it is carbon atom.Naphthenic base can be saturation or part is unsaturated.Naphthenic base can be connect with aromatic ring, tie point On the carbon atom of non-aromatic ring carbon atom.Naphthenic base includes the ring member nitrogen atoms from 3 to 10.In certain specific aspects, naphthenic base choosing From in cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, suberyl and cyclooctyl.Naphthenic base can be substituted or not replace.

Term " ester " refers to the chemical segment with-COOR group, and wherein R contains alkyl, naphthenic base, virtue selected from group Base, heteroaryl (being connected by ring carbon) (are connected) with heterolipid cyclic hydrocarbon by ring carbon.If desired, appointing in compound as described herein What hydroxy or carboxy side chain is esterified.The example of the process and specific group that prepare this kind of ester can refer to Greene and Wuts, Protective Groups in Organic Synthesis, 3^rdEd., John Wiley&Sons, New York, NY, 1999。

Term " halogen " or " halide " refer to fluorine, chlorine, bromine or iodine.

Term " miscellaneous alkyl " refer in alkyl one or more skeletal atoms selected from the atom other than carbon atom, such as oxygen, Nitrogen, sulphur, phosphorus or they exist simultaneously.On the one hand, miscellaneous alkyl C₁-C₆Miscellaneous alkyl.

Term " heterocycle " or " heterocycle " refer to hetero-aromatic ring (also referred to as heteroaryl) and Heterocyclylalkyl (also referred to as heterolipid ring Group), one to four hetero atom is contained in ring, wherein each hetero atom is selected from O, S and N, each heterocycle in ring system Containing 4 to 10 atoms, but any ring cannot contain that there are two neighbouring O or S atom.Non-aromatic heterocyclic groups are (also referred to as miscellaneous Naphthenic base) it include only having the group of 3 atoms in ring, but aryl-heterocyclic base must have at least five atom in ring.It is miscellaneous Ring group includes benzo ring system.The example of ternary heterocycle is aziridinyl；The example of quaternary heterocycle is azelidinyl；Five yuan The example of heterocycle is thiazolyl；The example of six-membered heterocyclic group is pyridyl group；The example of ten circle heterocyclic ring bases is quinolyl.It is non-aromatic The example of heterocycle is pyrrolidinyl, tetrahydrofuran base, dihydrofuryl, tetrahydro-thienyl, oxazolidine ketone group, oxinane Base, dihydro pyranyl, tetrahydro thiapyran base, piperidyl, morpholinyl, thiomorpholine base, thiophene alkyl, piperazinyl, aziridinyl, azacyclo- Butyl, oxygen cyclobutyl, thietanyl, class piperidyl, oxepanyl, thiepanyl, oxazepinyl, diazepine base, Thiazepinyl, 1,2,3,6- tetrahydro pyridyl, pyrrolin -2- base, pyrrolin -3- base, indoline base, 2H- pyranose, 4H- Pyranose, dioxanes base, 1,3-dioxolane, pyrazolinyl, dithianyl, dithiolane base, dihydro pyranyl, dihydro-thiophene Base, dihydrofuryl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3- azabicyclic [3.1.0] hexyl, 3- azabicyclic [4.1.0] heptane base, 3H- indyl and quinazinyl.The example of aromatic heterocyclic be pyridyl group, imidazole radicals, pyrimidine radicals, pyrazolyl, Triazolyl, pyrazinyl, tetrazole radical, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrole radicals, quinolyl, Isoquinolyl, indyl, benzimidazolyl, benzofuranyl, cinnolines, indazolyl, indolizine base, phthalazinyl, pyridazinyl, three Piperazine base, isoindolyl, pteridyl, purine radicals, di azoly, thiadiazole base, furazanyl, benzofuraxan base, benzothienyl, benzene Benzothiazolyl, benzoxazolyl group, quinazolyl, quinoxalinyl, naphthyridines base, furandipine base (furopyridinyl).Aforementioned group It can be connection carbon or connection nitrogen.For example, can be pyrroles -1- base (connection N) or pyrroles -3- from the group of pyrrole derivatives Base (connection C).Further, imidazoles -1- base or imidazo-3-yl (being all connected with N) or imidazoles-be can be from the group of imidazole derivatives 2- base, imidazol-4 yl or imidazoles -5- base (being all connected with C).Heterocycle includes benzo fusion ring system.Nonaromatic heterocycles can be by one Replaced a or two oxygen (=O) segments, such as pyrroline-2-one.

Term " heteroaryl " or " aromatic heterocycle " refer to including one or more the change of the ring hetero atom of oxygen and sulphur selected from nitrogen Close object.More preferably, heteroaryl includes indoles, azaindole, pyrroles, pyrazoles, pyrimidine, pyrazine, pyridine, quinoline, thiophene and furans. On the one hand, a heteroaryl includes 0 to 3 nitrogen-atoms.On the other hand, heteroaryl includes 0 to 3 nitrogen-atoms, and 0 to 1 Oxygen atom and 0 to 1 sulphur atom.On the other hand, heteroaryl is monocycle or bicyclic heteroaryl.

" Heterocyclylalkyl ", " heteroalicyclyl " or " nonaromatic heterocycles " refer to that a naphthenic base is contained at least one selected from nitrogen, The hetero atom of oxygen and sulphur.Free radical can be merged with aryl or heteroaryl.In certain specific aspects, Heterocyclylalkyl is selected from oxazolidine Base, pyrrolidinyl, tetrahydrofuran base, tetrahydro-thienyl, THP trtrahydropyranyl, tetrahydro thiapyran base, piperidyl, morpholinyl, thiomorpholine Base, piperazinyl and indoline base.Term heteroalicyclyl also includes all carbohydrate rings, including but not limited to monosaccharide, disaccharides and low Glycan.On the one hand, Heterocyclylalkyl is C₂-C₁₀Heterocyclylalkyl.On the other hand, Heterocyclylalkyl is C₄-C₁₀Heterocyclylalkyl.On the one hand, Heterocyclylalkyl contains 0 to 2 nitrogen-atoms.On the other hand, Heterocyclylalkyl contains 0 to 2 nitrogen-atoms, 0 to 2 oxygen atoms or 0 to 1 A sulphur atom.

Term " key " refers between two atoms or between two segments (when the atom connected by key is considered as big structure When a part) chemical bond.On the one hand, when group as described herein is a key, lack with reference to group, allow remaining It determines and forms a key between group.

Term " member ring " includes any cyclic structure.Term " member " means the quantity for indicating to constitute the skeletal atom of ring.This Sample, e.g., cyclohexyl, pyridyl group, pyranose, thiapyran base is hexatomic ring, and cyclopenta, pyrrole radicals, furyl and thienyl are five yuan Ring.

Term " optionally replacing " or " substitution " refer to can be replaced one or more additional groups with reference to group, additional base Group individually and it is independent be selected from, alkyl, naphthenic base, aryl, heteroaryl, heterolipid cyclic hydrocarbon, hydroxyl, alkoxy, alkylthio group, virtue Sulfenyl, alkane sulfoxide group, fragrant sulfoxide group, alkane sulfuryl, fragrant sulfuryl, cyano, halogen, carbonyl, thiocarbonyl, nitro, alkylhalide group, fluothane Base and amino, including monosubstituted and disubstituted amino group and its protected derivative.It can be with for example, optionally replacing It is halide ,-CN ,-NO₂Or L_sR_s, wherein each L_sIt is independent to be selected from a key ,-O- ,-C (=O)-,-C (=O) O- ,- S- ,-S (=O)-,-S (=O)₂,-NH- ,-NHC (=O)-,-C (=O) NH-, S (=O)₂NH- ,-NHS (=O)₂,-OC (= O) NH- ,-NHC (=O) O-, or-(C₁-C₆Alkyl)；Each R_sSelected from hydrogen, alkyl, fluoroalkyl, miscellaneous alkyl, naphthenic base, virtue Base, heteroaryl or Heterocyclylalkyl.Can be formed the protection derivative of the above substituent group protecting group can with reference to Greene and Wuts.On the one hand, optional substituent group is selected from halogen, CF₃, OH, CN, NO₂, SO₃H, SO₂NH₂, SO₂Me, NH₂, COOH, CONH₂, Alkoxy ,-N (CH₃)₂And alkyl.

In certain specific embodiments, the compound has one or more stereocenters, and each center is with R or S Type is individually present.Compound includes all diastereomeric bodily forms as mentioned herein, and the mapping bodily form is poor appropriate with theirs to the structure bodily form Mixture.Stereoisomer can be obtained by method of such as chiral chromatographic column to Enantiomer separation.

Method described herein and molecular formula include using N- oxide (if appropriate), and crystal form is (also referred to as more Crystal form) or formula (I) structural compounds pharmaceutically acceptable salt, with the active metabolism with identical these active compounds Object.In some cases, compound may exist as tautomer.All tautomers include as mentioned herein Within the scope of compound.In some specific embodiment, the compound exists with solvation form, pharmaceutically acceptable Solvent such as water, ethyl alcohol etc..In other specific embodiments, the compound exists with nonsolvated forms.

Specific pharmacy and medical terminology

Term " acceptable " refers to that a prescription component or active constituent are good for general treatment target as used herein The inexcessive adverse effect of health.

Term " Btk dependence ", as used herein, referring to may not cause in the case where lacking Btk, or will not cause phase With the imbalance or symptom of degree.

Term " Btk mediation " refers to disease or symptom caused in the presence of Btk, when Btk is lacked as used herein There may be similar situations.

Term " cancer " refers to a kind of misgrowth of out of contior cell, and under certain conditions as used herein (propagation) can be shifted.Such cancer includes but is not limited to, solid tumor (such as bladder, intestines, brain, chest, uterus, heart, Kidney, lung, lymphoid tissue (lymthoma), ovary, pancreas or other endocrine organs (such as thyroid gland), prostate, skin (melanin Tumor) or blood tumor (such as aleukemic leukemia).

Term " administering drug combinations " or its similar terms refer to several selected therapeutic agents to a disease as used herein People's medication, with identical or different administration mode in identical or different time administration.

Term " enhancing " or " can enhance " refer to that expected result can be in potency or duration side as used herein There are increase or extension in face.Therefore, in terms of the therapeutic effect of enhancing drug, term " can enhance " refers to that drug mentions in systems High or extension potency or the ability of duration." synergism value " used herein, refers in ideal system, can be to greatest extent The ability of another therapeutic agent of the enhancing on ground.

Term " inflammatory disease ", which refers to, one or more following illness features.Such as pain, fever, rubescent, enlargement, temporary Or permanent afunction.Inflammation has many forms of expression, including but not limited to, acute, sticky, atrophic, Catarrhal, slow Property, hardening, diffusivity, diffusive, exudative, fibre-bearing, at fibroid, part, granuloma, Hypertrophic, hypertrophy, interstitial Property, metastatic, gangrenosum acne, occlusive, substance, plasticity, generate property, proliferative, it is pseudomembranosa, suppurative, cause hardenability, slurry Liquid fibrin, blood plasma, pure, specificity, it is subacute, urge purulence, be toxicity, traumatic, and/or exedens.More into One step, inflammatory disease further includes, but is not limited to following blood vessel presentation (panarteritis, temporal arteritis)；Joint presentation (joint It is scorching --- crystallinity, psoriasis, reactivity, rheumatic, relies characteristic at bone)；Gastrointestinal tract presentation (colitis)；Skin (skin It is scorching)；Or the presentation (systemic lupus erythematosus) of Various Tissues and organ.

Term " immunity disease " refers to bad or adverse reaction disease or disease to endogenous or exogenous antigen generation Shape.As a result it usually will cause the dysfunction of cell or therefore destroy and cause dysfunction or destroy that there may be immune The organ or tissue of symptom.

" metabolite " of compound as used herein refers to the derivative generated during compound metabolism.Term is " living Property metabolin " refers to that is generated during compound metabolism has the derivative of biological activity.Term " metabolism ", such as this paper institute It states, refers to the whole process (including but not limited to, hydrolysis and the reaction by enzymatic) that predetermined substance is converted by organism.Cause This, enzyme can specifically change the structure of compound.Such as, cytochrome P 540 is catalyzed a series of oxidations and reduction reaction, and urinates The glucuronic acid molecules of glycosides diphosphonic acid glucuronyl transferase catalytic activation be connected to aryl alcohol, aliphatic alcohol, carboxylic acid, amine and from By on sulfydryl.Metabolin used herein, which refers to, gives compound to a kind of host, then analyzes the tissue sample of host；Or Compound is incubated for altogether in vitro with liver cell, then the compound after analysis reaction.

Term " subject " or " patient " include mammal and nonmammalian.Mammal includes but is not limited to feed Newborn class: people, non-human primates such as orangutan, ape and monkey class；Agricultural animal such as ox, horse, goat, sheep, pig；Domestic animal such as rabbit, dog；It is real Testing animal includes rodent, such as rat, mouse and cavy.Non-mammalian animal includes but is not limited to bird, fish etc..It is excellent one It selects in example, selected mammal is people.

Term " treatment ", " therapeutic process " or " therapy " as used herein, including mitigation, inhibition or improve disease symptom Or situation；Inhibit the generation of complication；Improve or prevent potential metabolic syndrome；Inhibit the generation of disease or symptom, such as controls The development of disease or situation；Mitigate disease or symptom；Disease or symptom is set to decline；Mitigate concurrent as caused by disease or symptom Disease, or prevention or the treatment sign as caused by disease or symptom.

As used herein, a certain compound or pharmaceutical composition after administration, can be such that a certain disease, symptom or situation obtains To improvement, espespecially its severity is improved, delayed onset, slows down disease progression, or reduce the state of an illness duration.No matter fix Administration or interim administration are administered continuously or interrupted continuous administration, can be attributed to or the situation related with administration.

Compound

In one aspect of the invention, a kind of compound of formula I or its pharmaceutically acceptable salt, solvate, work are provided Property metabolin, polymorph, ester, isomers or prodrug:

Wherein, L₁It is the alkenyl of alkyl, unsubstituted or substituted C2-C6 selected from unsubstituted or substituted C1-C6, unsubstituted Or the alkynyl of the C2-C6 replaced；

L₂Selected from-(CH₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n- NH-, wherein n is 0,1,2,3 or 4；

Ar is selected from unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl；

The alkyl of the unsubstituted or substituted C1-C6 indicates the linear chain or branched chain saturated hydrocarbons for being unsubstituted or being substituted Base, the linear chain or branched chain group including 1 to 6 carbon atom, including but not limited to methyl, ethyl, n-propyl, isopropyl, cyclopropyl Base, normal-butyl, cyclobutyl, isobutyl group, tert-butyl, n-pentyl, isopentyl or cyclopenta；The unsubstituted or substituted C2-C6 Alkenyl indicate the linear chain or branched chain alkenyl that is unsubstituted or is substituted, the linear chain or branched chain group including 2 to 6 carbon atoms, The including but not limited to ethylene, propylene, 1- butylene of cis or trans, 2- butylene, 1,3-butadiene, 1- amylene, 2- amylene, 1,3- Pentadiene, 1,4- pentadiene, 1- alkene, 2- alkene, 3- alkene, 1,3- hexadiene, 1,4- hexadiene, 1,5- hexadiene, 2,4- Hexadiene, 1,3,5- hexatriene；The alkynyl of the unsubstituted or substituted C2-C6 indicates the straight chain with one or more three keys Or branch alkynyl, including but not limited to acetylene, propine, 1- butine, 2- butine, 1,3- diacetylene, 1- pentyne, valerylene, 1,3- Pentadiine, 1,4- pentadiine, 1- alkynes, 2- alkynes, 3- alkynes, 1,3- hexadiyne, 1,4- hexadiyne, 1,5- hexadiyne, 2,4- Hexadiyne, 1,3,5- three alkynes.

Wherein, alkyl described above, alkenyl, alkynyl, aryl, heteroaryl and-(CH₂)_nIt is optional one or more Replaced various substituent groups in following: hydroxyl, halogen ,-C_1-4Alkyl and-OC_1-4Alkyl；The halogen is selected from fluorine, chlorine, bromine Or iodine.

In certain specific embodiments, L₁C2-C6 alkenyl selected from unsubstituted or substituted cis or trans and unsubstituted Or the C2-C6 alkynyl replaced.

In certain specific embodiments, L₁C2 alkenyl selected from unsubstituted or substituted cis or trans and unsubstituted or take The C2 alkynyl in generation, L₂Selected from-(CH₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n- NH-, wherein n is 0,1,2,3 or 4； Further preferred n is 1 or 2.

In certain specific embodiments, L₁C2 alkenyl selected from unsubstituted or substituted cis or trans and unsubstituted or take The C2 alkynyl in generation, L₂Selected from-(CH₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n- NH-, wherein n is 1 or 2.Ar is selected from Unsubstituted or substituted aryl and unsubstituted or substituted heteroaryl.

In certain specific embodiments, L₁C2 alkenyl selected from unsubstituted or substituted cis or trans and unsubstituted or take The C2 alkynyl in generation, L₂Selected from-(CH₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n- NH-, wherein n is 1 or 2.Ar is selected from Unsubstituted or substituted aryl.

The pyrazolo as shown in formula I [3,4-d] pyrimidines, wherein Y is selected from unsubstituted or substituted alkyl radical, or Person is 4 yuan, 5 yuan, 6 yuan of cycloalkyl rings；And R₁Selected from H or low alkyl group；

Alternatively, Y and R₁One 4 yuan, 5 yuan or 6 yuan of heterocycle can be combined together to form；

G is selected from H,

Wherein, R₂、R₃And R₄Separately selected from H, halogen ,-COOH, unsubstituted or substituted low alkyl group, unsubstituted Or the Lower heteroalkyl replaced；

In certain specific embodiments,It is selected from

More in certain specific embodiments,It is selected from

Also more in certain specific embodiments,It is selected from

In certain specific embodiments, G is

In certain specific embodiments, G isWherein R₂Selected from H ,-COOH and optionally by following base The low alkyl group that group replaces: halogen ,-OH ,-O- low alkyl group, amino, alkyl monosubstituted amino, dialkyl amido, Heterocyclylalkyl ammonia Base, alkanoyl oxygroup, alkyl sulfonyl amino；

In certain specific embodiments, G isWherein R₂Selected from H and low alkyl group.

In other specific embodiments, G is selected from

Any combination of group described above for different variables all gives expection herein.

Formula (I) compound includes but is not limited to the explanation in table 1.

Table 1.

The synthesis of compound

The method combined in the synthetic technology or well known technology and text of the usable standard of formula (I) compound described above To synthesize.In addition, solvent, temperature and other reaction conditions can change as mentioned herein.

The starting material of synthesis for formula (I) compound can be obtained by synthesizing or from commercial source, e.g., but it is unlimited In Aldrich Chemical Co. (Milwaukee, Wis.) or Sigma Chemical Co. (St.Louis, Mo.).Herein The compound has related compounds that well known technology and raw material can be used to synthesize with other with different substituents, including It is found in March, ADVANCED ORGANIC CHEMISTRY 4^thEd., (Wiley 1992)；Carey and Sundberg, ADVANCED ORGANIC CHEMISTRY 4^thEd., Vols.A and B (Plenum 2000,2001), Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3^rdEd., the method in (Wiley 1999).Compound preparation Conventional method not isoplastic condition can be introduced by using reagent appropriate and provided herein in molecular formula to change.

Formula (I) compound as described herein, in some embodiments, can with the synthesis of synthetic route shown in following scheme Compound as described herein is prepared by following methods.Following methods and embodiment are in order to illustrate these methods.These processes Limitation of the present invention is not construed in any way as with embodiment.Standard well known by persons skilled in the art can also be used Synthetic technology synthesizes compound described herein, or means known in the art and methods described herein are applied in combination.

The synthetic method of compound I includes the following steps:

The synthesis of step A:I-c: in a solvent, under the action of inorganic base, compound I-a and I-b replace anti- It answers, obtains compound I-c；

Wherein, L in the structure of compound I-a₁It is alkyl selected from unsubstituted or substituted C1-C6, unsubstituted or substituted The alkynyl of the alkenyl of C2-C6 and unsubstituted or substituted C2-C6；-(CH₂)_nIn n be 0,1,2,3 or 4；X is selected from halogen；Change It closes Ar in the structure of object I-b and is selected from unsubstituted or substituted aryl and unsubstituted or substituted heteroaryl；R₂Selected from hydroxyl, amino Or sulfydryl；L in the structure of compound I-c₂Selected from-(CH₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n- NH-, wherein n It is 0,1,2,3 or 4；Preferably, L₁C2 alkenyl and unsubstituted or substituted C2 selected from unsubstituted or substituted cis or trans Alkynyl；-(CH₂)_nIn n be 0,1 or 2；X is selected from bromine；Ar is selected from unsubstituted or substituted aryl；R₂Selected from hydroxyl；L₂Selected from- (CH₂)_n- O-, wherein n is 0,1 or 2.

The method of prepare compound I-c can be the conventional method of such substitution reaction in this field, in the present invention especially It is preferred that following reaction method and condition: compound I-b being mixed with organic solvent and inorganic base, and then is occurred with compound I-a Compound I-c is made in substitution reaction；

In the method for prepare compound I-c, the preferred ether solvent of the solvent, alcohols solvent, nitrile solvents and third One of ketone, DMF, water are a variety of；Further preferred acetone and DMF.The preferred tetrahydrofuran of the ether solvent.Described The preferred methanol of alcohols solvent and/or ethyl alcohol.The preferred acetonitrile of the nitrile solvents.

In the method for prepare compound I-c, the volume mass of the solvent and the compound I-b are than preferred 5mL/g-50mL/g, further preferred 10mL/g-30mL/g.

In the method for prepare compound I-c, the preferred potassium hydroxide of the inorganic base, potassium carbonate, sodium hydroxide, carbonic acid One of sodium and cesium carbonate are a variety of, further preferred potassium carbonate.

In the method for prepare compound I-c, the preferred 1:1- of molar ratio of the inorganic base and the compound I-b 5:1, further preferred 1:1-2:1.

In the method for prepare compound I-c, the molar ratio of the compound I-a and the compound I-b are preferred 1:1-5:1, further preferred 1:1-2:1.

In the method for prepare compound I-c, preferably 0 DEG C -150 DEG C of the temperature of the substitution reaction, further preferably 30℃-70℃。

In the method for prepare compound I-c, the process of the substitution reaction can be surveyed by the routine in this field Method for testing (such as TLC or HPLC) is monitored, and as the terminal of reaction when generally being disappeared using compound I-b, the reaction time is preferred 0.5h-16h, further preferred 3h-10h.

Following any post-processing step is preferably included in the method for prepare compound I-c: excellent when solvent is selected from DMF Choosing uses method (1).When solvent is selected from acetone, it is preferred to use method (2).Method (1) includes the following steps: that reaction terminates Afterwards, organic solvent is added and water, organic phase is washed with water and saturated sodium-chloride, dry, concentration, column chromatography.Method (2) packet It includes following steps: after reaction, filtering, concentration, column chromatography.Wherein, the further preferred acetic acid of the organic solvent Ethyl ester.The method and condition of the column chromatography is selected according to the method and condition that the column of this field routine chromatographs.

The synthesis of step B:I-e: in the presence of transition metal catalysts and alkalis, make compound I-c and compound I-d in Coupling reaction is carried out in the solvent not having a negative impact under nitrogen protection and to reaction with prepare compound I-e.

Wherein, L in the structure of compound I-c, compound I-d and compound I-e₁Selected from unsubstituted or substituted C1-C6 Alkyl, the alkenyl of unsubstituted or substituted C2-C6 and the alkynyl of unsubstituted or substituted C2-C6；L₂Selected from-(CH₂)_n-、- (CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n- NH-, wherein n is 0,1,2,3 or 4；Ar be selected from unsubstituted or substituted aryl and Unsubstituted or substituted heteroaryl；Y is selected from unsubstituted or substituted alkyl radical or 4 yuan, 5 yuan, 6 yuan of cycloalkyl rings；And R₁Choosing From H or low alkyl group；Alternatively, Y and R₁One 4 yuan, 5 yuan or 6 yuan of heterocycle can be combined together to form；Preferably, L₁C2 alkenyl and unsubstituted or substituted C2 alkynyl selected from unsubstituted or substituted cis or trans；L₂Selected from- (CH₂)_n- O-, wherein n is 0,1 or 2；Ar is selected from unsubstituted or substituted aryl；It is selected from It is highly preferred thatIt is selected from

The method of prepare compound I-e can be the conventional method of such coupling reaction in this field, in the present invention especially It is preferred that following reaction method and condition: under nitrogen protection, compound I-d and organic solvent, transition-metal catalyst, alkali being mixed It closes, and then reacts 3-15 hours (preferably 10 hours) generation coupling reactions under 30-150 DEG C (preferably 80 DEG C) with compound I-c Compound I-e is made；

In the method for prepare compound I-e, the solvent can be used not to reaction have a negative impact it is any Solvent.It preferably includes hydrocarbon solvent (such as benzene, toluene and dimethylbenzene), nitrile solvents, ether solvent (such as dimethoxy second Alkane, tetrahydrofuran and 1,4- dioxane), alcohols solvent, aprotic polar solvent (such as DMF, DMSO and hexamethyl phosphinylidyne Amine) and one of water or a variety of；Further preferred DMF.The preferred 1,4- dioxane of the ether solvent.The alcohols The preferred methanol of solvent and/or ethyl alcohol.The preferred acetonitrile of the nitrile solvents.

In the method for prepare compound I-e, the volume mass of the solvent and the compound I-d are than preferred 5mL/g-50mL/g, further preferred 10mL/g-30mL/g.

In the method for prepare compound I-e, the example of the alkali includes: organic base, such as triethylamine, diisopropyl Ethamine, pyridine, lutidines, trimethylpyridine, 4-dimethylaminopyridine, tertiary potassium butyrate, tertiary sodium butyrate, sodium methoxide, second Sodium alkoxide, lithium hexamethyldisilazide, sodium hexamethyldisilazide, potassium hexamethyldisilazide and butyl lithium；And nothing Machine alkali, such as sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide and sodium hydride.Further preferred triethylamine and two Wopropyl ethyl amine.

In the method for prepare compound I-e, the preferred 1:1-5:1 of molar ratio of the alkali and the compound I-d, Further preferred 1:1-2:1.

In the method for prepare compound I-e, the molar ratio of the compound I-c and the compound I-d are preferred 1:1-5:1, further preferred 3:1-5:1.

In the method for prepare compound I-e, the example that can be used for the transition-metal catalyst of the step includes that palladium is urged Agent (such as acid chloride, three (dibenzalacetone) two caesiums and 1,1 '-bis- (diphenylphosphino) ferrocene-palladium chlorides (II)- Dichloromethane complex etc.).If desired, ligand (such as triphenylphosphine and tri-tert-butylphosphine) can be added, and copper can be tried Agent (such as cuprous iodide and copper acetate) is used as co-catalyst.Further preferred transition-metal catalyst is four (triphenylphosphines) Palladium, co-catalyst are cuprous iodide.

In the method for prepare compound I-e, the molar ratio of the compound I-d and the transition-metal catalyst It is preferred that 0.0001-1, further preferred 0.01-0.5.The compound I-d and the molar ratio of the ligand is preferred 0.0001-4, further preferred 0.01-0.2.The compound I-d and the molar ratio of the co-catalyst is preferred 0.0001-4, further preferred 0.01-0.2.

In the method for prepare compound I-e, the temperature of the temperature of the coupling reaction preferably 0 DEG C-solvent refluxing, into - 150 DEG C of the preferred room temperature of one step.

In the method for prepare compound I-e, the process of the coupling reaction can be surveyed by the routine in this field Method for testing (such as TLC or HPLC) is monitored, and as the terminal of reaction when generally being disappeared using compound I-d, the reaction time is preferred 0.5h-20h, further preferred 3h-15h.

It, can be after separating and purifying with means of purification by known sharing in the method for prepare compound I-e Or subsequent step is carried out without separating and purifying with means of purification by known separation, it is described known Separation and means of purification are for example concentrated, are concentrated in vacuo, crystallizing, solvent extraction, precipitating and column chromatograph.

The method of prepare compound I-e preferably includes following post-processing step: after reaction, water and organic molten is added Agent, organic phase are washed through water and saturated sodium-chloride, and organic phase is dry with anhydrous sodium sulfate, are filtered, concentration, through column chromatographic purifying, ?.Wherein, the further preferred ethyl acetate of the organic solvent or methylene chloride.The method and condition of the column chromatography It is selected according to the method and condition that the column of this field routine chromatographs.

The synthesis of step C:I-f: in a solvent, under the action of an acid, compound I-e is obtained into chemical combination through deprotection reaction Object I-f；

Wherein, L in the structure of compound I-e and compound I-f₁Alkyl selected from unsubstituted or substituted C1-C6 does not take The alkynyl of the alkenyl and unsubstituted or substituted C2-C6 of generation or the C2-C6 replaced；L₂Selected from-(CH₂)_n-、-(CH₂)_n-O-、- (CH₂)_n-S-、-(CH₂)_n- NH-, wherein n is 0,1,2,3 or 4；Ar be selected from unsubstituted or substituted aryl and it is unsubstituted or Substituted heteroaryl；Y is selected from unsubstituted or substituted alkyl radical or 4 yuan, 5 yuan, 6 yuan of cycloalkyl rings；And R₁Selected from H Or low alkyl group；Alternatively, Y and R₁One 4 yuan, 5 yuan or 6 yuan of heterocycle can be combined together to form；Preferably, L₁ C2 alkenyl and unsubstituted or substituted C2 alkynyl selected from unsubstituted or substituted cis or trans；L₂Selected from- (CH₂)_n- O-, wherein n is 0,1 or 2；Ar is selected from unsubstituted or substituted aryl；It is selected from It is highly preferred thatIt is selected from

The method of prepare compound I-f can be the conventional method of such deprotection in this field, especially excellent in the present invention It selects following reaction method and condition: compound I-e and organic solvent and acid-mixed is closed, be deprotected and compound I-f is made；

In the method for prepare compound I-f, the preferred ether solvent of the solvent, alcohols solvent, nitrile solvents and third One of ketone, DMF, ethyl acetate, methylene chloride, water are a variety of；Further preferred ethyl acetate and methylene chloride.Described The preferred tetrahydrofuran of ether solvent.The preferred methanol of the alcohols solvent and/or ethyl alcohol.The preferred acetonitrile of the nitrile solvents.

In the method for prepare compound I-f, the volume mass of the solvent and the compound I-e are than preferred 5mL/g-50mL/g, further preferred 10mL/g-30mL/g.

In the method for prepare compound I-f, the acid preferably organic acid or inorganic acid, the organic acid are selected from first Acid, acetic acid, trifluoroacetic acid, malic acid, maleic acid, fumaric acid, succinic acid, benzoic acid, methanesulfonic acid, benzene sulfonic acid, acetic acid, propionic acid, Butyric acid, octanoic acid, adipic acid, ethanedioic acid, malonic acid, succinic acid, tartaric acid, phenylacetic acid, phthalic acid, terephthalic acid (TPA), penta Acid, caproic acid, capric acid, stearic acid, palmitic acid, acrylic acid etc.；The inorganic acid be hydrochloric acid, hydrobromic acid, phosphoric acid, hydrophosphate, Sulfuric acid or disulfate etc.., further preferred hydrochloric acid or trifluoroacetic acid.

In the method for prepare compound I-f, the preferred 1:1-20 of molar ratio of the acid and the compound I-e: 1, further preferred 5:1-10:1.

It is preferably 0 DEG C -150 DEG C of the temperature of the deprotection reaction, further excellent in the method for prepare compound I-f Select room temperature.

In the method for prepare compound I-f, the process of the deprotection reaction can pass through the routine in this field Test method (such as TLC or HPLC) is monitored, and as the terminal of reaction when generally being disappeared using compound I-e, the reaction time is excellent Select 0.5h-20h, further preferred 3h-12h.

The method of prepare compound I-f preferably includes following post-processing step: after reaction, organic solvent is added and beats Slurry filters, is dry.Wherein, the preferred alcohols solvent of the organic solvent, ether solvent, hydrocarbon solvent etc..Further Preferably ethyl alcohol and petroleum ether.

The synthesis of step D:I: in a solvent, under the action of alkali and/or condensing agent, by compound I-f and G-Z carry out at Amide reaction, obtains compound I；

Wherein, L in the structure of compound I-f and compound I₁It is alkyl selected from unsubstituted or substituted C1-C6, unsubstituted Or replace C2-C6 alkenyl and unsubstituted or substituted C2-C6 alkynyl；L₂Selected from-(CH₂)_n-、-(CH₂)_n-O-、- (CH₂)_n-S-、-(CH₂)_n- NH-, wherein n is 0,1,2,3 or 4；Ar is selected from unsubstituted or substituted aryl and unsubstituted or take The heteroaryl in generation；Y is selected from unsubstituted or substituted alkyl radical or 4 yuan, 5 yuan, 6 yuan of cycloalkyl rings；And R₁Selected from H or rudimentary Alkyl；Alternatively, Y and R₁One 4 yuan, 5 yuan or 6 yuan of heterocycle can be combined together to form；G is selected from H, Wherein, R₂、R₃And R₄Separately select From H, halogen ,-COOH, unsubstituted or substituted low alkyl group, unsubstituted or substituted Lower heteroalkyl；Z is selected from halogen or hydroxyl Base；Preferably, L₁C2 alkenyl and unsubstituted or substituted C2 alkynyl selected from unsubstituted or substituted cis or trans； L₂Selected from-(CH₂)_n- O-, wherein n is 0,1 or 2；Ar is selected from unsubstituted or substituted aryl；It is selected from It is highly preferred thatIt is selected fromG is selected from Z Selected from chlorine.

The conventional method that the method for prepare compound I can react for such in this field at amide, in the present invention especially It is preferred that following reaction method and condition: compound I-f being mixed with organic solvent and alkali and/or condensing agent, and then and compound G-Z (unsubstituted or substituted lower paraffin hydrocarbon, carboxylic acid or carboxylic acid halides) is reacted at amide is made compound I；

In the method for prepare compound I, the preferred ether solvent of the solvent, alcohols solvent, nitrile solvents, alkanes are molten One of agent and aprotic polar solvent are a variety of；Further preferred alkanes solvent.The preferred tetrahydro furan of the ether solvent It mutters.The preferred methanol of the alcohols solvent and/or ethyl alcohol.The preferred acetonitrile of the nitrile solvents.The alkanes solvent is preferred Methylene chloride.The preferred N,N-dimethylformamide of the aprotic polar solvent.

In the method for prepare compound I, the volume mass of the solvent and the compound I-f are than preferred 5mL/ G-50mL/g, further preferred 10mL/g-30mL/g.

In the method for prepare compound I, the preferred organic base of the alkali, such as triethylamine, diisopropylethylamine, pyrrole Pyridine, lutidines, trimethylpyridine, 4-dimethylaminopyridine, tertiary potassium butyrate, tertiary sodium butyrate, sodium methoxide, sodium ethoxide, six Two silicon substrate lithium amide of methyl, sodium hexamethyldisilazide, potassium hexamethyldisilazide and butyl lithium；And inorganic base, example Such as sodium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, sodium hydroxide and sodium hydride.Further preferred triethylamine and diisopropyl second Amine.

In the method for prepare compound I, the preferred 1:1-5:1 of molar ratio of the alkali and the compound I-f, into The preferred 2:1-3:1 of one step.

In the method for prepare compound I, the condensing agent preferably has CDI, DCC, EDCI, DIC, HATU, HBTU, PyBOP etc., if desired, acylation catalyst, such as DMAP, HOBt etc. can be added.Further preferred EDCI and HATU.

In the method for prepare compound I, the preferred 1:1-5 of molar ratio of the condensing agent and the compound G-Z: 1, further preferred 1:1-3:1.

In the method for prepare compound I, the molar ratio of the acylation catalyst and the compound G-Z is preferred 0.001:1-5:1, further preferred 0.1:1-2:1.

In the method for prepare compound I, the molar ratio preferably 1 of the compound G-Z and the compound I-f: 1-5:1, further preferred 1:1-3:1.

In the method for prepare compound I, preferably -30 DEG C -150 DEG C of the temperature at amide reaction is further excellent - 10 DEG C -30 DEG C of choosing.

In the method for prepare compound I, the process at amide reaction can be surveyed by the routine in this field Method for testing (such as TLC or HPLC) is monitored, and as the terminal of reaction when generally being disappeared using compound I-f, the reaction time is preferred 0.1h-30h, further preferred 0.1h-16h.

Following any post-processing step is preferably included in the method for prepare compound I: when Z is selected from chlorine, it is preferred to use side Method (1).When Z is selected from hydroxyl, it is preferred to use method (2).Method (1) includes the following steps: after reaction, water to be added, has Machine mutually uses water, aqueous citric acid solution, saturated sodium bicarbonate and saturated sodium-chloride washing, dry, concentration, column chromatography.Method (2) include the following steps: after reaction, organic solvent to be added and water, organic phase are washed with water and saturated sodium-chloride, it is dry, Concentration, column chromatography.Wherein, the further preferred ethyl acetate of the organic solvent or methylene chloride.The column chromatography Method and condition according to this field routine column chromatograph method and condition selected.

Step E: the synthesis of compound 2: in a solvent, compound 1 and NIS is subjected to substitution reaction, obtain compound 2；

The method of prepare compound 2 can be the conventional method of such substitution reaction in this field, especially excellent in the present invention It selects following reaction method and condition: compound 1 is mixed with organic solvent, and then substitution reaction occurs with NIS, compound is made 2；

In the method for prepare compound 2, the preferred ether solvent of the solvent, alcohols solvent, nitrile solvents and acetone, One of DMF, water are a variety of；Further preferred DMF.The preferred tetrahydrofuran of the ether solvent.The alcohols solvent It is preferred that methanol and/or ethyl alcohol.The preferred acetonitrile of the nitrile solvents.

In the method for prepare compound 2, the volume mass of the solvent and the compound 1 is than preferred 5mL/g- 50mL/g, further preferred 10mL/g-30mL/g.

In the method for prepare compound 2, the preferred 1:1-1:5 of molar ratio of the compound 1 and the NIS, into The preferred 1:1-1:2 of one step.

In the method for prepare compound 2, preferably 0 DEG C -150 DEG C of the temperature of the substitution reaction, further preferred 50 ℃-100℃。

In the method for prepare compound 2, the process of the substitution reaction can pass through the routine test in this field Method (such as TLC or HPLC) is monitored, as the terminal of reaction, reaction time preferred 0.5h- when generally being disappeared using compound 1 16h, further preferred 5h-12h.

In the method for prepare compound 2 when solvent is selected from DMF preferably following post-processing step: be down to room temperature, be added Water, solid filtering, is successively washed with water and organic solvent, 50-90 DEG C of drying.Wherein, the organic solvent is further excellent Select ethyl alcohol.

The synthesis of step F:I-d: in a solvent, in the presence of three replace phosphine and disubstituted azodicarboxylate, chemical combination Object 2 occurs Mitsunobu with compound 3 and reacts.

Wherein, the Y in compound 3 and compound I-d is selected from unsubstituted or substituted alkyl radical, either 4 yuan, 5 yuan, 6 yuan of cycloalkyl rings；And R₁Selected from H or low alkyl group；Alternatively, Y and R₁It can combine It is formed together one 4 yuan, 5 yuan or 6 yuan of heterocycle；Preferably,It is selected from It is highly preferred thatIt is selected from

The method of prepare compound I-d can be the conventional method of such Mitsunobu reaction in this field, in the present invention Particularly preferably following reaction method and condition: in a solvent, in the presence of triphenylphosphine and diisopropyl azodiformate, chemical combination Object 2 occurs Mitsunobu with compound 3 and reacts.

The method of prepare compound I-d preferably includes following steps: at 0 DEG C -10 DEG C, by diisopropyl azodiformate It instills in the solution that compound 2, triphenylphosphine and compound 3 and solvent are formed, carries out Mitsunobu reaction.

In the method for prepare compound I-d, the preferred nitrile solvents of the solvent, ether solvent and halogenated hydrocarbon solvent One of or it is a variety of；Further preferred ether solvent.The preferred acetonitrile of the nitrile solvents；The ether solvent preferably four Hydrogen furans；The preferred chlorinated hydrocarbon solvent of the halogenated hydrocarbon solvent, further preferred methylene chloride.

In the method for prepare compound I-d, the volume mass of the solvent and the compound 2 is than preferred 1mL/ G-40mL/g, further preferred 5mL/g-20mL/g.

In the method for prepare compound I-d, mole of the diisopropyl azodiformate and the compound 2 Than preferred 1:1-3:1, further preferred 1:1-2:1.

In the method for prepare compound I-d, the preferred 1:1- of molar ratio of the triphenylphosphine and the compound 2 3:1, further preferred 1:1-2:1.

In the method three of prepare compound I-d, the molar ratio preferably 1 of the compound 3 and the compound 2: 1-3:1, further preferred 1:1-2:1.

In the method for prepare compound I-d, preferably -10 DEG C -35 DEG C of the temperature of the Mitsunobu reaction, into one Preferably 0 DEG C -20 DEG C of step.

In the method for prepare compound I-d, the process of the Mitsunobu reaction can be by normal in this field Rule test method (such as TLC or HPLC) are monitored, as the terminal of reaction, reaction time when generally being disappeared using compound I-d It is preferred that 0.5h-24h, further preferred 3h-12h.

The method of prepare compound I-d preferably includes following post-processing step: after reaction, evaporating solvent under reduced pressure, warp Column chromatographic purifying.The method and condition of the described column chromatography according to the column chromatography of this field routine method and condition into Row selection.

The synthesis of formula (I) compound is summarized in embodiment.

The further form of compound

In some specific embodiment, formula (I) compound according to pharmaceutically acceptable acid-addition salts, (pharmaceutically may be used by one kind The salt of receiving) it prepares, free alkali form and pharmaceutically acceptable inorganic or organic acid reaction by compound, including but It is not limited to inorganic acid, such as hydrochloric acid, hydrobromic acid, phosphoric acid, hydrophosphate, sulfuric acid, disulfate etc.；Organic acid formic acid, acetic acid, trifluoro Acetic acid, malic acid, maleic acid, fumaric acid, succinic acid, benzoic acid, methanesulfonic acid, benzene sulfonic acid, acetic acid, propionic acid, butyric acid, octanoic acid, oneself Diacid, ethanedioic acid, malonic acid, succinic acid, tartaric acid, phenylacetic acid, phthalic acid, terephthalic acid (TPA), valeric acid, caproic acid, capric acid, Stearic acid, palmitic acid, acrylic acid etc..

Inorganic acid preferably is selected from hydrochloric acid, hydrobromic acid or phosphoric acid.Organic acid preferably be selected from formic acid, acetic acid, trifluoroacetic acid, malic acid, Maleic acid, fumaric acid, succinic acid, benzoic acid, methanesulfonic acid or benzene sulfonic acid；More preferably from acetic acid or trifluoroacetic acid.

" pharmaceutically acceptable " herein refers to a kind of substance, such as carrier or dilution, will not make compound bioactivity or Property disappears, and relative nontoxic e.g. gives individual something, will not cause undesired biotic influence or in harmful manner It interacts with any component that it contains.

Term " pharmaceutically acceptable salt " refers to that a kind of existence form of compound, the form will not cause organic to being administered The important stimulation of body, and the bioactivity of compound and property will not be made to disappear.It is pharmaceutically acceptable in certain specific aspects Salt be that acquisition, such as hydrochloric acid are reacted with acid by formula (I) compound, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, P-methyl benzenesulfonic acid, salicylic acid etc..Pharmaceutically acceptable salt can also react forming salt, such as ammonium salt with alkali by formula (I) compound； Alkali metal salt, such as sodium or sylvite；Alkali salt, such as calcium or magnesium salts；Organic alkali salt, such as dicyclohexylamine, N- methyl D-glucose Amine, trihydroxymethylaminomethane；Amino-acid salt, such as arginine, lysine etc..

It should be understood that the reference of pharmaceutically acceptable salt includes solvent addition form or crystal form, especially solvate Or polymorphic.Solvate contains stoichiometry or non-stoichiometric solvent, and be with pharmaceutically acceptable solvent such as water, Ethyl alcohol etc., it is selectively formed in crystallisation procedure.Hydrate is formed when solvent is water, or alcohol is formed when solvent is ethyl alcohol Compound.The solvate of formula (I) compound is very easily made or is formed according to method described herein.For example, formula (I) hydrate of compound is recrystallized from water/organic solvent in the mixed solvent and is easily made, the organic solvent packet used It includes but is not limited to, dioxane, tetrahydrofuran, ethyl alcohol or methanol.In addition, compound can be with non-solvent as mentioned herein Change and solvation form exists.In short, solvation form is considered suitable for the purpose of for Compounds and methods for provided herein In nonsolvated forms.

In other specific embodiments, formula (I) compound is prepared to different forms, including but not limited to, amorphous, Crush shape and millimicro-granularity form.In addition, formula (I) compound includes crystal type, polymorphic can also be used as.Polymorphic includes The different crystalline lattice arrangement of the identical element composition of compound.Polymorphic usually has different x-ray diffraction patterns, and infrared spectroscopy melts Point, density, hardness, crystal form, the property of light and electricity, stability and dissolubility.Different factors such as recrystallization solvent, crystalline rate It is leading for may cause single crystal form with storage temperature.

In certain specific embodiments, formula (I) compound is prepared as prodrug." prodrug " refers to that a reagent turns in vivo Turn to prototype medicine.Prodrug is usually useful, because under certain conditions, they may administration easier than prototype medicine.They can With, for example, be administered orally and can be with biological utilisation, but prototype medicine is not all right.Prodrug can also improve original on pharmaceutical composition The dissolubility of type medicine.For example, there is no limit prodrug is formula (I) compound, is unfavorable for the case where passing through cell membrane in water solubility Under, prodrug makes to be easier by cell membrane as ester administration, is then hydrolyzed into carboxylic acid by metabolism, active entities once enter It is water-soluble just highly beneficial in cell.Further example, prodrug can be a small peptide (polyaminoacid) and be connected to one Acid groups, peptide show active fragment after being metabolized.

Prodrug is usually the precursor of medicine, and next administration and absorption are converted into active material, or pass through some processes Become the stronger type of activity, is such as converted by metabolic pathway.The chemical group that some prodrugs have make its activity it is lower and/or Compare the dissolubility or some other properties of prototype medicine.Once the chemical group of prodrug is removed and/or modifies it, lived Property medicine.Prodrug be usually it is useful, in some cases, their administrations easier than prototype medicine.In certain specific embodiment, this Prodrug compound described in text is administered orally and can be with biological utilisation, but prototype medicine is not all right.Moreover, in certain specific implementations In example, prodrug as described herein can also improve the dissolubility of prototype medicine on pharmaceutical composition.

In other specific embodiments, prodrug is designed as reversible medicaments derivative, as modifier come using with Tissue of the enhancing medicament transport to specific location.At specific aspect, it is that water is that the purpose of prodrug design, which is for target area, It is effectively water-soluble can to increase it for the therapeutic compounds of primary solvent.Fedorak et al., Am.J.Physiol., 269: G210-218(1995)；McLoed et al., Gastroenterol, 106:405-413 (1994)；Hochhaus et al., Biomed.Chrom., (1992) 6:283-286；J.Larsen and H.Bundgaard, Int.J.Pharmaceutics, 37,87 (1987)；J.Larsen et al., Int.J.Pharmaceutics, 47,103 (1988)；Sinkula et al., J.Pharm.Sci., (1975) 64:181-210；T.Higuchi and V.Stella, Prodrugs as Novel Delivery Systems, Vol.14 of the A.C.S.Symposium Series；With Edward B.Roche, Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987.

In another specific embodiment, compound as described herein has isotope labelling (e.g., radioisotope) Or by other methods, including but not limited to, using chromophore or fluorescence segment, feel cold signal or chemiluminescent labeling.

On the other hand, formula (I) compound has one or more Stereocenters, and each center is independently deposited with R or S configuration ?.Compound includes all non-corresponding isomers as mentioned herein, enantiomer, epimer and they properly mix object. In some specific aspect, formula (I) compound as their single stereoisomers, preparation can by make racemic mixture with Optical activity resolution reagent reacts to form a pair of of diastereoisomer, and separation diastereoisomer obtains optically pure enantiomer. In certain specific aspects, the separation of enantiomer is come using the covalent diastereoisomer derivative of compound as described herein It carries out.In other specific aspects, use separable compound (such as crystallization diastereomeric salt).Diastereoisomer has not Same physical property (e.g., fusing point, boiling point, dissolubility, reactivity etc.), and in specific aspect, diastereoisomer passes through benefit It is separated with these different characteristics.In these specific aspects, diastereoisomer is by chiral chromatogram or is based on dissolubility Different isolation technics separates.By any practicable method that not will lead to racemization, together with resolution reagent, Obtain optically pure enantiomer.Jean Jacques, Andre Collet, Samuel H.Wilen, " Enantiomers, Racemates and Resolutions ", John Wiley And Sons, Inc., 1981.

In addition, compound exists as geometric isomer provided herein in a certain specific embodiment.Provided herein Compounds and methods for includes all cis-, trans-, E formula, and Z formula isomers properly mixes object with they.In certain specific aspects, Compound as described herein exists as tautomer.All tautomers molecular structural formula range as described herein It is interior.The another aspect of Compounds and methods for provided herein, from single preparation step (in conjunction with or interconversion) obtained enantiomer And/or the mixture of diastereomer is expected.

Therapeutical uses

On the one hand, compound of formula I can inhibit Btk.On the other hand, compound of formula I shows antiproliferative activity and can be effective Treat proliferative diseases in ground.On the one hand, compound of formula I show prevention or treatment inflammation, it is relevant to aberrant B cell proliferation oneself The effect of body immunity disease and tumor disease.

In further aspect, it provided herein is a kind of containing therapeutically effective amount by giving curer in need At least one compound of formula I or composition, to inhibit the active method of the bruton's tyrosine kinase of the subject.? In some embodiments, subject's suffering from autoimmune disease in need, such as inflammatory bowel disease, arthritis, lupus, class Rheumatic arthritis, psoriasis arthropathica, osteoarthritis, Still disease, adolescent arthritis, rheumatoid arthritis are comprehensive It seeks peace oneself immunity hepatitis etc..

In further embodiment, subject in need suffers from cancer.In one embodiment, the cancer Disease is B cell proliferative disease, for example, diffusivity large B cell lymphoid tumor, follicular lymphoma, chronic lymphocytic leukemia, Chronic lymphocytic leukemia, B cell pre-lymphocytic leukemia, lymphoplasmacytic lymphoma/huge ball of Walden Si Telun Proteinemia, splenic marginal zone lymthoma, plasma cell myeloma, plasmacytoma, extranodal marginal zone B cell lymphoma, lymph node Marginal zone B-cell lymphoma, mantle cell lymphoma, vertical diaphragm (thymus gland) large B cell lymphoid tumor, intravascular large B cell lymphoma, Lymphoma primary effusion, Burkitt lymphoma/leukaemia or lymphomatoid granulomatosis etc..

The present invention also provides compound or pharmaceutically acceptable salt thereofs of the present invention to prepare the application in Btk inhibitor, special It is not the application in preparation treatment cell proliferative diseases.The cell proliferative diseases include cancer.In other words, the present invention is gone back It provides compound described in general formula I or its pharmaceutically acceptable salt or solvate individually or is used in combination with other drugs Treat the application in proliferative diseases (such as cancer).It can be used in combination with compound or pharmaceutically acceptable salt thereof provided by the present invention Antineoplastic include but and the non-limiting following type of at least one: mitotic inhibitor (such as vincaleukoblastinum, eldisine and Changchun Rui Bin)；Tubulin decomposing inhibitor (such as taxol)；Alkylating reagent (such as cis-platinum, carboplatin and cyclophosphamide)；Antimetabolite (such as 5 FU 5 fluorouracil, Tegafur, methotrexate (MTX), cytarabine and hydroxycarbamide)；Can be inserted into antibiotic, (such as A Leisu, mitogen is mould Element and bleomycin)；Enzyme (such as lucid asparagus adnosine deaminase)；Topoisomerase enzyme inhibitor (as relied on primary glycosides and camptothecine) and biological respinse Regulator (such as interferon).

The purposes of compound is to prepare preparation in the present invention, comprising: directly uses compound or obtains during the preparation process Any ingredient use.The present invention also provides a kind of pharmaceutical composition, which includes that a certain amount of energy inhibits The compound of formula I of Btk activity and pharmaceutically acceptable excipient.The pharmaceutical composition is with aqueous dispersion, liquid, Gel Mile, syrup, elixir, medicine slurry, suspending agent, inhalant, controlled release agent, quick-dissolving agent, effervescent agent, freeze-dried, tablet, pulvis, pill, Dragee, capsule, sustained release agent, extended release agent, pulse release agent, multiparticulates agent or the form of releasing agent at once.

Pharmaceutical composition set forth herein is by pharmaceutically acceptable diluent, excipient or bonding agent and compound of formula I Or its pharmaceutically acceptable salt, its pharmaceutically acceptable prodrug or its pharmaceutically acceptable solvate composition.

On the one hand, pharmaceutical composition set forth herein contains certain a effective amount of compound as described above and pharmaceutically Acceptable excipient.On the other hand, the pharmaceutical composition in addition to containing other than compound of formula I, include second pharmaceutically Active ingredient.

In a particular embodiment, a kind of pharmaceutical composition is provided, which includes: i) physiologically acceptable Carrier, diluent and/or excipient；And ii) a kind of compound set forth herein.

Any of the above-described aspect contains further embodiment, including gives certain a effective amount of compound of formula I merely.This A little preferences specifically include: i) disposably giving compound of formula I；Ii mammal compound of formula I) is repeatedly given in one day； Iii compound of formula I) is persistently given；Or iv) continuously give compound of formula I.

Any of the above-described aspect contains further embodiment, including repeatedly gives certain a effective amount of compound of formula I.This A little preferences specifically include: i) single dose compound of formula I；Ii) multiple dosing, time interval are six hours；Iii) every Give mammal compound of formula I within eight hours.In further or optional preference, this method has drug holiday, specifically Refer to compound of formula I gives temporary delay or the dosage of compound of formula I is by temporary reduction；Terminate in drug holiday When, restore the dosage of compound of formula I.The time of drug holiday year is differed from two days to one.

Medicine group can be made with pharmaceutically various typical additives (such as diluent and excipient) in the compound of the present invention Close object.According to therapeutic purposes, pharmaceutical composition can be made to various types of administration unit dosage forms, as tablet, pill, pulvis, Liquid, suspension, lotion, granule, capsule, suppository and injection (solution and suspension) etc..

In order to shape the pharmaceutical composition of tablet form, it can be used this field any of and widely used figuration Agent.For example, carrier, such as lactose, white sugar, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, avicel cellulose and silicon Acid etc.；Adhesive, such as water, ethyl alcohol, propyl alcohol, common syrup, glucose solution, starch solution, penetrating judgment solution, carboxymethyl cellulose Element, lac, methylcellulose and potassium phosphate, polyvinylpyrrolidone etc.；Disintegrating agent, such as dried starch, mosanom, agar powder and sea Band powder, sodium bicarbonate, calcium carbonate, the aliphatic ester of polyethylene sorbitan, lauryl sodium sulfate, stearic acid monoglycerides, Starch and lactose etc.；Disintegration inhibitor, such as white sugar, glycerol tristearate, coconut oil and hydrogenated oil and fat；Adsorption enhancer, such as season Amine base and lauryl sodium sulfate etc.；Wetting agent, such as glycerol, starch；Adsorbent, such as starch, lactose, kaolin, bentonite With colloid silicic acid etc.；And lubricant, such as pure talcum, stearate, boric acid powder and polyethylene glycol etc..If necessary Words can also use common coated material to make tablet as sugar coated tablet, painting gelatin film tablet, enteric coated tablets, film coated tablets, double Tunic tablet and multilayer tablet.

In order to shape the pharmaceutical composition of pill, it can be used this field any of and widely used inborn nature Agent, for example, carrier, such as lactose, starch, coconut oil, hardened vegetable oils, kaolin and talcum etc.；Adhesive, such as gum arabic Powder, Huang write rubber powder, gelatin and ethyl alcohol etc.；Disintegrating agent, such as agar and Kelp Powder.

In order to shape the pharmaceutical composition of suppository form, it can be used this field any known and widely used inborn nature Agent, for example, polyethylene glycol, coconut oil, higher alcohol, the ester of higher alcohol, gelatin and semi-synthetic glyceride etc..

, can be by solution and suspension liquid disinfectant in order to prepare the pharmaceutical composition of injection form, and it is preferably added suitable chlorine Change sodium, glucose or glycerol etc. are made and the isotonic injection of blood.When preparing injection, it is possible to use in the art any Common carrier.For example, water, ethyl alcohol, propylene glycol, the isooctadecanol of ethoxylation, the isooctadecanol and polyethylene of polyoxy The aliphatic ester etc. of anhydro sorbitol.In addition, common lytic agent, buffer and analgesic etc. can also be added.As needed, exist During treating schizophrenia, colorant, preservative, fragrance, flavoring agent, sweetening agent and other medicines etc. can also be added.

The content of compound and its pharmaceutically acceptable salt shown in formula I of the invention in pharmaceutical composition is without spy Different limitation, can be selected in a wide range, generally can be mass percent 1-90%, preferably mass percent 1- 30%.

In the present invention, the medication of the pharmaceutical composition is not particularly limited.Can according to patient age, gender and Other conditions and symptom select the preparation of various dosage forms to be administered.For example, tablet, pill, solution, suspension, lotion, granule It is oral administration with capsule；Injection can be administered alone, or (such as glucose solution and amino acid are molten with injection conveying liquid Liquid) it is mixed into row vein injection, muscle can be carried out with injection merely if necessary, inject in intradermal, subcutaneous or abdomen；Suppository is It is administered into rectum.

In the present invention, use can be properly selected according to method of administration, patient age, gender and other conditions and symptom Pharmaceutical quantities.

The positive effect of the present invention is that:

Pyrazolo [3,4-d] pyrimidines and its salt of the invention have good inhibitory effect, partization to Btk It closes object activity/water solubility and replaces Buddhist nun according to Shandong better than marketed drug, provide diversification in styles for the research and development of Btk micromolecular inhibitor Structure has the good prospect of marketing.

Without prejudice to the field on the basis of common sense, above-mentioned each optimum condition, can any combination to get the present invention it is each preferably Example.

The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to the reality It applies among a range.In the following examples, the experimental methods for specific conditions are not specified, according to conventional methods and conditions, or according to quotient It is selected with specification.Although without being described in further detail, it is believed that those skilled in the art can retouching based on this paper It states, the disclosure is fully utilized.

In the examples below, undefined abbreviation has its generally accepted meaning, unless stated otherwise.Table 1, table 2 It is respectively physicochemical data, the compound I-1-I-24 of compound I-1-I-24 with table 3¹H-NMR data and I-1-I-24 are external Enzyme level inhibitory activity data.

Embodiment 1

The preparation of key intermediate 4

Step A: the synthesis of compound 2

By compound 1 (15g, 111mmol), NIS (30g, 133.2mmol) and DMF (150ml) in 90 DEG C of stirring 4h, after It is down to room temperature, water 150ml is added, solid filtering successively uses water 100ml, ethyl alcohol 100ml to wash, and 70 DEG C of dry solid 23g are received Rate: 79.4%.

Step B: the synthesis of compound 4

By compound 2 (15g, 57.47mmol), (S) -1- tertbutyloxycarbonyl -3- hydroxy piperidine (23.2g, 114.9mmol), triphenylphosphine (30.1g, 114.9mmol) and 150ml THF are cooled to 0 DEG C in ice salt bath, and DIAD is added dropwise (23.2g, 114.9mmol), is warmed to room temperature and is stirred overnight, and column chromatographs to obtain solid 13.1g, yield: 51.3%.

Embodiment 2

The preparation of compound I-1

Step A: the synthesis of compound 5

Propargyl bromide (18.9g, 159mmol), phenol (10g, 106mmol), potassium carbonate (44g, 318mmol) is in 100mlDMF In be stirred overnight at room temperature, be added ethyl acetate, successively use saturated sodium bicarbonate, saturated sodium-chloride, water washing, after concentration it is transparent Oily liquids 11.9g, yield 84.3%.

Step B: the synthesis of compound 6

By compound 4 (4.45g, 10mmol), CuI (0.38g, 2mmol), Pd (PPh₃)₄(1.39g, 1.2mmol), Et₃N (2.03g, 20mmol), compound 5 (6.62g, 50mmol) and 50ml DMF are warming up to 80 DEG C under nitrogen and are stirred overnight, and are down to Room temperature, rear evaporating column chromatograph to obtain solid 3.31g, yield: 73.6%.

Step C: the synthesis of compound 7

Compound 6 (3.3g, 10.5mmol), 4M HCl-1,4-dioxane (20ml, 72mmol) and 20ml ethyl acetate, It is stirred overnight at room temperature, dries to obtain solid 1.9g, yield: 67.1% for 75 DEG C after filtering.

Step D: the synthesis of compound I-1

By compound 7 (600mg, 1.56mmol), DIPEA (403mg, 3.12mmol) and 30mlDCM are cooling in ice salt bath, The DCM solution 10ml of acryloyl chloride (183mg, 2.03mmol) is added dropwise, is warmed to room temperature, concentration rear pillar chromatographs to obtain solid 420mg, receives Rate: 67%.

Embodiment 3

The preparation of compound I-2

The synthesis of compound I-2 is completed by using the step of being similar to described in embodiment 2.

Embodiment 4

The preparation of compound I-3

The synthesis of compound I-3 is completed by using the step of being similar to described in embodiment 2.

Embodiment 5

The preparation of compound I-4

The synthesis of compound I-4 is completed by using the step of being similar to described in embodiment 2.

Embodiment 6

The preparation of compound I-5

The synthesis of compound I-5 is completed by using the step of being similar to described in embodiment 2.

Embodiment 7

The preparation of compound I-6

The synthesis of compound I-6 is completed by using the step of being similar to described in embodiment 2.

Embodiment 8

The preparation of compound I-7

The synthesis of compound I-7 is completed by using the step of being similar to described in embodiment 2.

Embodiment 9

The preparation of compound I-8

The synthesis of compound I-8 is completed by using the step of being similar to described in embodiment 2.

Embodiment 10

The preparation of compound I-9

The synthesis of compound I-9 is completed by using the step of being similar to described in embodiment 2.

Embodiment 11

The preparation of compound I-10

The synthesis of compound I-10 is completed by using the step of being similar to described in embodiment 2.

Embodiment 12

The preparation of compound I-11

The synthesis of compound I-11 is completed by using the step of being similar to described in embodiment 2.

Embodiment 13

The preparation of compound I-12

The synthesis of compound I-12 is completed by using the step of being similar to described in embodiment 2.

Embodiment 14

The preparation of compound I-13

The synthesis of compound I-13 is completed by using the step of being similar to described in embodiment 2.

Embodiment 15

The preparation of compound I-14

The synthesis of compound I-14 is completed by using the step of being similar to described in embodiment 2.

Embodiment 16

The preparation of compound I-15

The synthesis of compound I-15 is completed by using the step of being similar to described in embodiment 2.

Embodiment 17

The preparation of compound I-16

Step A: the synthesis of compound 8

By 3- butyne-1-ol (10g, 142.67mmol), phenol (16.11g, 171.2mmol), triphenylphosphine (44.9g, 171.2mmol) and 100mlTHF is cooling in ice salt bath, 0 DEG C of dropwise addition DIAD (34.62g, 171.2mmol), after be warmed to room temperature stirring Overnight, liquid 5.45g, yield: 26.1% are chromatographed to obtain through column.

Step B: the synthesis of compound 9,10 and I-16 are completed by using the step of being similar to described in embodiment 2.

Embodiment 18

The preparation of compound I-17

Step A: the synthesis of compound 11

By compound 7 (880mg, 2.3mmol), 10%Pd-BaSO4 (260mg, 0.23mmol) and 20ml methanol are in normal pressure Lower 45 DEG C of hydrogen are stirred overnight, and rear pillar chromatographs to obtain solid 470mg, yield: 60%.

Step B: the synthesis of compound I-17 is completed by using the step of being similar to described in embodiment 2.

Embodiment 19

The preparation of compound I-18

Step A: the synthesis of compound 12

LiAlH is added at 0 DEG C in compound 7 (1.57g, 4.08mmol) and 70ml THF₄(929mg, 24.47mmol), it is warmed to room temperature and is stirred overnight, ethyl acetate is added and sodium hydrate aqueous solution, rear pillar chromatograph to obtain solid 330mg, Yield: 23.2%.

Step B: the synthesis of compound I-18 is completed by using the step of being similar to described in embodiment 2.

Embodiment 20

The preparation of compound I-19

Step A: the synthesis of compound 13

By compound 7 (1g, 2.6mmol), 10%Pd/C (400mg) and 30ml methanol are stirred under atmospheric hydrogen in 55 DEG C Overnight, filtering rear pillar chromatographs to obtain grease 890mg, yield: 88.08%.

Step B: the synthesis of compound I-19 is completed by using the step of being similar to described in embodiment 2.

Embodiment 21

The preparation of compound I-20

Step A: the synthesis of compound 14

I-PrMgCl (2M in is added in trimethyl acetenyl silicon (23g, 233.88mmol) and 100ml THF at 0 DEG C THF 117ml, 233.88mmol) stirring 30min, be warmed to room temperature stirring 30min, be added CuBr (5.03g, 35.08mmol) and Benzyl bromine (10g, 58.47mmol) back flow reaction 5h is down to room temperature, and ethyl acetate, washing is added, and rear pillar chromatographs to obtain liquid 9.2g, Yield: 83.5%.

Step B: the synthesis of compound 15

Compound 14 (9.2g, 48.85mmol) and 100ml methanol is cooling in ice bath, addition potassium carbonate (33.76g, 2h 244.2mmol) is stirred, is stirred overnight at room temperature, ethyl acetate is added, washing obtains grease 5g, direct plunges into next after concentration Step.

Step C: the synthesis of compound 16,17 and I-20 are completed by using the step of being similar to described in embodiment 2.

Embodiment 22

The preparation of compound I-21

2- perfluoroalkyl acrylate (175mg, 1.95mmol) and 10ml DMF are added at 0 DEG C DIPEA (504mg, 3.9mmol), HATU (741mg, 1.95mmol) and compound 7 (500mg, 1.3mmol), are stirred overnight at room temperature, and acetic acid second is added Ester, washing, rear pillar chromatograph to obtain solid 330mg, yield: 60%.

Embodiment 23

The preparation of compound I-22

Step A: the synthesis of compound 18 is completed by using step B described in embodiment 1 is similar to.

Step B: the synthesis of compound 19,20 and I-22 are completed by using the step of being similar to described in embodiment 2.

Embodiment 24

The preparation of compound I-23

Step A: the synthesis of compound 21 is completed by using step B described in embodiment 1 is similar to.

Step B: the synthesis of compound 22,23 and I-23 are completed by using the step of being similar to described in embodiment 2.

Embodiment 25

The preparation of compound I-24

Step A: the synthesis of compound 24 is completed by using step B described in embodiment 1 is similar to.

Step B: the synthesis of compound 25,26 and I-24 are completed by using the step of being similar to described in embodiment 2.

Heretofore described room temperature refers to environment temperature, is 15 DEG C -30 DEG C.

The reagents and materials used in the present invention are commercially available.

2 compound I-1-I-24 physicochemical data of table

3 compound I-1-I-24's of table¹H-NMR data

Embodiment 26

The Btk inhibitory activity of Compound ira vitro biochemistry enzyme level is tested

Principle and method:

The kinase activity detection for establishing Btk micromolecular inhibitor using homogeneous phase time discrimination fluorescence (HTRF) method is flat Platform carries out the active measurement of compound enzyme level.Btk inhibitor activity determination method is can be with single phosphorus of TK class kinase reaction The other end of polyadenylation sites substrate connects biotin, at the end biotin, biotin energy and streptavidin-XL665 (acceptor) it is connected, completes label.And by after phosphorylation, which just can be with the antibody of anti-phosphorylation site ( Indicate Doner) it combines, to generate FRET signal, as shown in Figure 1.

Operating procedure:

1. untested compound is dissolved in 100% DMSO, initial dilution concentration is respectively 100 μM, then in 96 orifice plates Middle configured buffer gradient dilutes.

2. the compound that 4 μ l of transfer have diluted is into 384 orifice plates.

3. preparing the buffer of substrate and ATP.

4. in the buffer to 384 orifice plates for adding 4 μ l substrates and ATP using microwell plate sample injector.

5. in the buffer to 384 orifice plates for adding 2 μ l enzymes using microwell plate sample injector.

6. gently shaking 384 orifice plates, the 1000g room temperature short time is centrifuged, and is incubated at room temperature 60 minutes.

7. preparing terminate liquid, each 5 μ l of SA-XL665 and TK antibody is separately added into microwell plate sample injector, shakes 1 minute, so After be centrifuged, be incubated at room temperature 60 minutes.Referring to attached drawing 2.

The multi-functional plate reader reading of 8.PHERAStar.

5.0 analysis software data of 9.GraphPad Prism.

Column formula acquires IC₅₀:

Calculate IC₅₀Value:

With log [administration concentration] for abscissa, inhibiting rate is ordinate, fits one in GraphPad Prism 5.0 Dose-effect curve, obtains drug concentration when its 50% inhibiting rate, i.e., IC of the compound in enzyme level thus₅₀Value.

The compound Btk inhibitory activity data are as follows:

Compound number	Btk IC50(nM)	Number	Btk IC50(nM)
				I-1	2.95	I-14	209.2
I-2	18.1	I-15	45.35
				I-3	3.95	I-16	39.8
I-4	15.9	I-17	23.2
				I-5	>500	I-18	11.3
I-6	>500	I-19	22.18
				I-7	49.23	I-20	102.4
I-8	20.85	I-21	>500
				I-9	26.89	I-22	>1000
I-10	38.17	I-23	>1000
				I-11	3.84	I-24	>1000
I-12	89.62	Ibrutinib	5.49
				I-13	18.11

The result shows that synthesized compound has excellent Btk inhibitory activity.Such as I- of part of compounds disclosed in it 1, the activity of I-3 and I-11 is better than Ibrutinb, to the Btk micromolecular inhibitor and structure of modification of Future Development structure diversification Play directive function.

Embodiment 27

The Btk inhibitory activity comparison of external biochemistry enzyme level

The Compound A structure formula that Chinese patent application CN103958512A is recorded are as follows:

The compound be FGFR inhibitor, bibliography CN103958512A report method synthesized compound A according to, The method of embodiment 26 tests the external BTK enzyme level inhibitory activity of compound A, as a result as follows:

Compound number	Btk IC50(nM)
		A	>1000

The result shows that compound provided by the invention inhibits the activity of Btk to be far more than compound A.

Embodiment 28

The BTK inhibitory activity of Compound ira vitro cellular level is tested

Method: CCK-8 detection method

Operating procedure:

1. preparing the cell suspension (hole 2000-3000cells/) of 100ul in 96 well culture plates, culture plate is being cultivated Case (37 DEG C, 5%CO₂) preculture 24 hours.

2. the untested compound of gradient concentration is added to culture plate.

3. culture plate is incubated for 48 hours in incubator.

4. 10ul CCK-8 solution is added to every hole, culture plate is gently shaken after addition.

5. culture plate is incubated for 1-4 hours in incubator.

6. measuring the absorbance at 450nm with microplate reader.

Record result:

Inhibitory rate of cell growth %=(control group absorbance value-experimental group absorbance value)/control group absorbance value * 100%；

Calculate IC₅₀Value:

With log [administration concentration] for abscissa, inhibiting rate is ordinate, fits one in GraphPad Prism 5.0 Dose-effect curve, obtains drug concentration when its 50% inhibiting rate, i.e., IC of the compound in cellular level thus₅₀Value.

Test Activity Results such as following table is such as shown:

Remarks: Ramos and Raji is typical bone-marrow-derived lymphocyte leukaemia cell, the expression of BTK kinases height.

The result shows that majority of compounds provided by the invention has lymphocytic leukemia cell (Ramos and Raji) Stronger inhibiting effect, wherein compound I-1 is shown, for the comparable cellular level inhibitory activity of Buddhist nun, indicates such with according to Shandong Molecule has exploitation at the potentiality of new and effective BTK inhibitor, to treatment-related tumor disease especially diffusivity large B cell Lymthoma or chronic lymphocytic leukemia have biggish application value.

Embodiment 29

The cLog P value of part of compounds

CLog P value is calculated by ACD-Labs (V6.0).

Compound	cLog P
		I-1	1.84±1.49
I-3	2.41±1.49
		I-4	2.42±1.49
I-11	1.64±1.49
		I-18	1.53±1.14
Buddhist nun (Ibrutinib) is replaced according to Shandong	2.92±1.18

The result shows that the dissolubility of compound provided by the invention in water is better than replacing Buddhist nun according to Shandong, so as to reduce medication Amount mitigates side effect.

Embodiment described herein is served only for explanation (illustratively), and the various modifications or change that technical staff is done also are answered Including in the spirit and scope of patent application and within accessory claim scope.

Claims (15)

1. a kind of compound of formula I or its pharmaceutically acceptable salt, solvate, active metabolite, polymorph, ester, isomery Body or prodrug:

Wherein:

L₁It is the alkenyl of alkyl, unsubstituted or substituted C2-C6 selected from unsubstituted or substituted C1-C6, unsubstituted or substituted The alkynyl of C2-C6；

L₂Selected from-(CH₂)_n-、-(CH₂)_n-O-、-(CH₂)_n-S-、-(CH₂)_n- NH-, wherein n is 0,1,2,3 or 4；

Ar is selected from unsubstituted or substituted aryl or heteroaryl；

Y is selected from unsubstituted or substituted alkyl radical or 4 yuan, 5 yuan, 6 yuan of cycloalkyl rings；

R₁Selected from H or low alkyl group；

Alternatively, Y and N and R₁It is connected to form a quaternary, five yuan or hexa-member heterocycle；

G is selected from H,

Wherein, R₂、R₃And R₄Separately selected from H, halogen ,-COOH, unsubstituted or substituted low alkyl group, unsubstituted or take The Lower heteroalkyl in generation.

2. compound as described in claim 1, in which:

N is 1 or 2.

3. compound as described in claim 1, in which:

Ar is selected from unsubstituted or substituted aryl.

4. compound as claimed in claim 3, in which: be substituted by meta or para position substitution.

5. compound as claimed in claim 4, in which: substituent group is alkoxy or halogen.

6. compound as described in claim 1, in which:

L₁The alkynyl of alkenyl or unsubstituted C2-C6 selected from unsubstituted C2-C6.

7. compound as claimed in claim 6, in which:

L₁Selected from vinyl or acetenyl.

8. compound as described in claim 1, in which:

Y and N and R₁It is connected to form hexa-member heterocycle.

9. compound as described in claim 1, in which:

G is

10. a kind of compound, which is characterized in that be selected from:

11. such as the described in any item compounds of claim 1-10, which is characterized in that the compound swashs bruton's tyrosine Enzyme (Bruton ' s tyrosine kinase, Btk) has inhibitory activity.

12. a kind of pharmaceutical composition, which includes the described in any item compounds of claim 1-11 and pharmaceutically acceptable Excipient.

13. pharmaceutical composition as claimed in claim 12, wherein the form of pharmaceutical composition is aqueous dispersion, liquid, Gel Mile, syrup, elixir, medicine slurry, suspension, aerosol, controlled release agent, quick-dissolving agent, effervescent agent, freeze-dried, tablet, powder, pill, Sugar-coat is complete, capsule, sustained release agent, extended release agent, pulse controlled release agent, multiparticulates agent or releases immediately agent.

14. a kind of preparation method of such as described in any item compounds of claim 1-11, which is characterized in that the method includes Step: under nitrogen protection, structure such as I-d compound represented of formula is mixed with DMF, palladium catalyst and organic base, Jin Eryu Structure such as I-c compound represented of formula are reacted 3-15 hours at 30-150 DEG C, obtained structure chemical combination as shown in I-e of formula Object is deprotected preparation structure intermediate as shown in I-f of formula in acid condition, then deposits in alkali or condensing agent and alkanes solvent Under, with G-Z carry out it is amide condensed react, obtain such as the described in any item compounds of claim 1-11；

Wherein: L₁、L₂、Ar、Y、R₁, G definition as shown in claim 1；Z is selected from halogen or hydroxyl.

15. a kind of increase such as the described in any item compounds of claim 1-11 in preparation prevention or treatment inflammation, with aberrant B cell Grow the application in relevant autoimmune disease (such as rheumatoid arthritis) and/or the drug of tumor disease.