CN109305944A - The inhibitor of bruton's tyrosine kinase - Google Patents
The inhibitor of bruton's tyrosine kinase Download PDFInfo
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- CN109305944A CN109305944A CN201710630341.0A CN201710630341A CN109305944A CN 109305944 A CN109305944 A CN 109305944A CN 201710630341 A CN201710630341 A CN 201710630341A CN 109305944 A CN109305944 A CN 109305944A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
- C07D239/70—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings condensed with carbocyclic rings or ring systems
- C07D239/72—Quinazolines; Hydrogenated quinazolines
- C07D239/78—Quinazolines; Hydrogenated quinazolines with hetero atoms directly attached in position 2
- C07D239/84—Nitrogen atoms
Abstract
The invention discloses a kind of formula (I) compound with Btk inhibitory activity, wherein all variables are as defined herein.The compound can be used for treating autoimmune disease, heteroimmunity disease, cancer or thromboembolic disorders.Also disclose the Pharmaceutical composition comprising formula (I) compound.The present invention provides can be in a manner of covalent bond, for example covalently irreversibly to inhibit the active compound of bruton's tyrosine kinase.
Description
Technical field
The present invention relates to inhibit Bu Ludun (Bruton ' s) tyrosine kinase (Btk) active compound, particularly, this hair
It is bright to further relate to covalently irreversibly to inhibit the active compound of bruton's tyrosine kinase.The compound of the present invention can be used
In the compound for treating disease related with B cell.
Background technique
" molecular targeted therapy " is using some key enzymes for participating in tumor cell differentiation breeding as drug screening target
The method that point carrys out treating cancer, has become the key areas of current cancer drug development.In recent years, protein tyrosine kinase
The research of (protein tyrosine kinase, PTK) signal path has become hot spot, and people are to PTKs and tumour cell
Proliferation, differentiation, migration and apoptosis relationship had a more understanding, interference or block the strategy of tyrosine kinase access
For the treatment of tumour, therefore the PTKs inhibitor for researching and developing high inhibitory activity has become a weight for developing new anti-tumor drug
Want method.
In PTK, bruton's tyrosine kinase (Bruton ' s tyrosine kinase, Btk) belongs to non-receptor type junket
A member of histidine kinase Tec family.Btk is in connection cell surface B-cell receptor (B-cell receptor, BCR) stimulation under
It swims in the B cell signal transduction path of intracellular response and plays the part of vital role, be B cell development, activation, signal transduction
With the key regulators of survival.Therefore and the relevant many human diseases of B cell are all related with the overexpression of Btk.Research card
It is bright by blocking the running of tyrosine kinase tumoricidal can transmit, to achieve the purpose that inhibit tumour, thus Btk
Inhibitor is the treatment method for effectively B cell being prevented to mediate.Clinically it is mainly used to treat the chain gamma globulin of X-
Mass formed by blood stasis (XLA), chronic lymphocytic leukemia (CLL), the huge B cell lymphoma of diffusivity etc., in addition have research confirm, Btk or from
The therapy target of body immunity disease, heteroimmunity disease, inflammatory disease and thromboembolic disorders.
Wherein, most successful Btk inhibitor is according to Shandong by one of present inventor exploitation for Buddhist nun (ibrutinib)
(referring to document 1 and 2).The drug is set to " breakthrough " new drug by FDA, and research and development have a extensive future.However it is more to still need to exploitation
Btk inhibitor.
Some in present inventor report a kind of (aminophenyiamino) pyrimidine in patent application before this
Yl-benzamide class inhibitor (referring to document 3), is proven to have Btk inhibitory activity.
In addition, thering is Gefitinib, Lip river in distress to replace at present as the protein tyrosine kinase inhibitor with quinazoline structure
The drugs such as Buddhist nun, Lapatinib, but their mainly EGF-R ELISA (EGFR) tyrosine kinase for being directed to, and its bone
Frame structure is 4- anilinoquinazoline substantially.In addition, being disclosed in document 4 a kind of using amido quinazoline as the chemical combination of skeleton
Object, but its Lck kinases for being directed to Src family.
In the above context, it is still necessary to which exploitation is more directed to the high-drug-effect inhibitor of Btk, furthermore, it is desirable to covalent bond
Mode inhibits kinases (especially bruton's tyrosine kinase) active compound.
Existing technical literature
Document 1:CN101610676A
Document 2:Zhengying Pan etc., Chem Med Chem 2007,2,58-61
Document 3:WO2013060098A1
Document 4:Erin F.Dimauro etc., J.Med.Chem.2006,49,5671-5686
Summary of the invention
Subject to be solved by the invention
The issue of the present invention is to provide a kind of efficient kinase inhibitor compounds for Btk.Further, of the invention
It also provides in a manner of covalent bond, especially covalently irreversibly inhibit kinases (especially bruton's tyrosine kinase) activity
Compound.
Means for solving the problems
In one aspect of the invention, it provides
A kind of compound of formula (I) or its pharmaceutically can or its pharmaceutically acceptable salt:
Wherein,
W is selected from H, C1-6Alkyl ,-(NH-CO)n-L-L3,-(CO-NH)n-L-L3,
Wherein,
L is key, C1-3Alkylidene or C2-3Alkenylene,
L3For C3-8Naphthenic base is such as
Aryl such as phenyl, naphthalene, phenanthryl, anthryl, fluorenyl and indenyl or bicyclic heteroaryl such as furyl, thienyl, pyrroles
Base, imidazole radicals, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazole radical, thiadiazolyl group, pyridine
Base, pyrimidine radicals, pyridazinyl, pyrazinyl etc.,
Condensed hetero ring base is such as
Oxo condensed hetero ring base is such as
The C3-8Naphthenic base, aryl, bicyclic heteroaryl, condensed hetero ring base, oxo condensed hetero ring base is optionally by following substituent groups
Replace: halogen, amino, C1-6Alkyl, C1-6Alkoxy, halogenated C1-6Alkyl, heterocycle, C1-6Alkyl-heterocyclyl groups, C1-6Alkyl-is miscellaneous
Ring group-C1-3Alkyl, C1-6Alkyl-S (O)2, (C1-6Alkyl)2P (O or S)-,
N is 0 or 1,
X is selected from H, halogen and C1-6Alkyl,
R1And R2Independently of one another, in the same manner or differently, H, C (O) and S (O) are selected from2；
L1And L2Independently of one another, in the same manner or differently, selected from for optionally by C1-3Alkyl-substituted C2-3Alkenyl；
On condition that working as R1When for H, L1It is not present, and works as R2When for H, L2It is not present.
In a preferred embodiment, W is selected from-(NH-CO)n-L-L3,-(CO-NH)n-L-L3,
Wherein,
L is key,
L3To be optionally selected from F, Cl, Br, amino, methyl, methoxyl group, ethyoxyl, propoxyl group, CF by 1,2 or 33,CH3-S(O)2, (CH3)2The cyclopropyl that the substituent group of P (O or S)-replaces,
Phenyl, naphthalene, furyl, thienyl, pyrrole radicals, thiazolyl, imidazole radicals, pyrazolyl, pyridyl group, pyrimidine radicals, pyrazinyl, pyridazine
Base, benzheterocycle base, pyrazolo heterocycle, oxo benzheterocycle base,
N is 1.
In another preferred embodiment, above-mentioned benzheterocycle base is nitrogen-containing benzoheterocycle base, and above-mentioned pyrazolo is miscellaneous
Ring group is pyrazolo nitrogen heterocycle, and above-mentioned oxo benzheterocycle base is oxo nitrogen-containing benzoheterocycle base.
In another preferred embodiment, wherein the benzheterocycle base are as follows:The pyrazolo heterocycle isThe oxygen
It is for benzheterocycle base
In another preferred embodiment, wherein X is selected from H, F, Cl and methyl, the-N (R1-L1)(R2-L2)
For-NH2Or-NH-C (O)-CH=CH2。
In another preferred embodiment, L3To be optionally selected from F, Cl, Br, methyl, CF by 1,2 or 33,CH3-S(O)2, (CH3)2The phenyl that the substituent group of P (O)-replaces, 3- pyridine
Base, 4- pyridyl group,
In another aspect of the present invention, a kind of compound is provided, is selected from:
In another aspect of the present invention, the compound is further selected from:
The present invention provides the inhibitor of bruton's tyrosine kinase, and further, the present invention provides with covalent bond
Mode inhibits the active inhibitor compound of bruton's tyrosine kinase.Particularly, the present invention provides can be with Bu Ludun junket
Cysteine residues on histidine kinase form the compound of covalent bond.
In one aspect, the present invention provides covalently irreversibly to inhibit the active chemical combination of bruton's tyrosine kinase
Object.
In a preferred aspect, in the present invention, covalently irreversibly to inhibit bruton's tyrosine kinase active
Compound is selected from:
In another aspect of the present invention, a kind of Pharmaceutical composition is provided, it includes a effective amount of aforementioned present invention
Compound and pharmaceutically acceptable carrier or excipient.
In another aspect of the present invention, it provides the compound of the present invention or is prepared by its pharmaceutically acceptable salt
The purposes in drug for treating following disease or situation: autoimmune disease, heteroimmunity disease, inflammation disease,
Cancer, thromboembolic disorders.
The effect of invention
The present invention provides one kind using (aminophenyiamino) quinazoline as skeleton, the suppression of high-drug-effect kinases for Btk
Inhibitor compound, particularly, the compound of the present invention can inhibit bruton's tyrosine kinase active in a manner of covalent bond.Into
One step, the present invention also provides covalently irreversibly to inhibit the active compound of bruton's tyrosine kinase.
Specific embodiment
Unless otherwise defined, all scientific and technical terminologies used herein all have the skill with claimed theme fields
Art personnel are commonly understood by identical meaning.The definition of standard chemistry terms can be found in reference works, including Carey and
" ADVANCED ORGANIC CHEMISTRY " A volumes of the fourth edition (2000) of Sundberg and B volumes (2001), Plenum
Press, New York.
“C1-6Alkyl " refers to that carbon atom is the alkyl of 1-6, including methyl, ethyl, propyl, butyl, amyl and hexyl, packet
Include all possible isomeric form, such as n-propyl and isopropyl, normal-butyl, isobutyl group, sec-butyl and tert-butyl, etc..
“C1-6Alkyl " includes whole subranges contained therein, such as C1-2Alkyl, C1-3Alkyl, C1-4Alkyl, C1-5Alkyl, C2-5Alkane
Base, C3-5Alkyl, C4-5Alkyl, C3-4Alkyl, C3-5Alkyl and C4-5Alkyl."C1-3Alkylidene " includes methylene, ethylidene, Asia third
Base and isopropylidene."C2-3Alkenyl " includes vinyl (- CH=CH2), acrylic (- CH=CHCH3) and isopropenyl (- C
(CH3)=CH2).Aromatic group refers to that planar rings have the pi-electron system of delocalization and contain 4n+2 pi-electron, and wherein n is whole
Number.Aromatic group can be made of five, six, seven, eight, nine or more than nine annular atoms.Aromatic group, which can be, optionally to be replaced.
Aromatic group includes that " aryl " (annular atom is only made of carbon atom) and " heteroaryl " (annular atom is by carbon atom and selected from for example
The hetero atom of oxygen, sulphur and nitrogen is constituted)." aryl " and " heteroaryl " includes monocycle or condensed ring is polycyclic (shares adjacent annular atom
Pair ring) group.The example of " aryl " includes but is not limited to phenyl, naphthalene, phenanthryl, anthryl, fluorenyl and indenyl."C3-8Cycloalkanes
Base " refer to nonaromatic, only containing carbon and hydrogen and annular atom number be 3-8 carbon atom one or more cyclic groups,
And its can be saturation, part it is unsaturated or completely it is unsaturated.C3-8The example of naphthenic base includes following part:
Deng.
" bicyclic heteroaryl " refers to containing 1-4 selected from N, the heteroatomic 5-6 unit monocycle aromatic group of O, S, example packet
It includes: furyl, thienyl, pyrrole radicals, imidazole radicals, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl,
Tetrazole radical, thiadiazolyl group, pyridyl group, pyrimidine radicals, pyridazinyl, pyrazinyl etc..
Condensed hetero ring base refers to phenyl ring and heterocyclic fused or heterocycle and heterocyclic fused condensed group together, and wherein benzo is miscellaneous
Ring group refers to phenyl ring and heterocyclic fused obtained group, and pyrazolo heterocycle is pyrazoles and other heterocyclic fused obtained groups,
Nitrogen-containing benzoheterocycle base refers to that in benzheterocycle base, the heterocycle condensed with phenyl ring contains nitrogen-atoms.The example includes:
Oxo condensed hetero ring refer to have on the carbon atom of a certain heterocycle in above-mentioned condensed hetero ring one or more=O replace and
Obtained condensed hetero ring, when condensed hetero ring belongs to phenyl ring with heterocyclic fused situation, substituent group=O is located at heterocycle rather than on phenyl ring.
The example includes:
" halogen " refers to fluorine, chlorine, bromine and iodine."C1-6Alkoxy " refers to (C1-6Alkyl) O- group, wherein C1-6Alkyl is as originally
It is defined in text." halogenated C1-6Alkyl " refers to halogen-(C1-6Alkyl)-group, wherein C1-6Alkyl is as defined herein.Halogenated C1-6
Alkyl includes perhalogeno C1-6Alkyl, wherein C1-6Whole hydrogen atoms in alkyl are replaced by halogen, such as-CF3、-CH2CF3、-
CF2CF3、-CH2CH2CF3Deng.
Heterocycle, which refers to, is selected from nitrogen-atoms containing at least one, the heteroatomic monocycle of oxygen atom and/or sulphur atom, two rings or
Spirocyclic ring system group, each ring include 3,4,5,6,7 or 8 atoms, can be saturation or unsaturation, preferably be saturated.In fact
Example includes morpholinyl, piperazinyl, piperidyl, oxetanyl, nitrogen heterocyclic heptyl, aziridine base, diaza cycloheptyl
Alkane, 1,3- diaza-cyclohexane, Isosorbide-5-Nitrae-diaza-cyclohexane, pyrrolidinyl, hexahydro azepineBase, oxetanyl, tetrahydro
Furyl, tetrahydro-thienyl, oxazolidinyl, thiazolidinyl, isoxazolidinyl, THP trtrahydropyranyl, thio-morpholinyl, quininuclidinyl
And imidazolinyl.Heterocycle is connected to by any substitutive carbon atom contained by the group or any substitutive nitrogen-atoms
In the main structure of compound, it is connected in main structure preferably by nitrogen-atoms.
C1-6Alkyl-heterocyclyl groups refer to by C1-6Alkyl-substituted heterocycle, wherein being connected to compound by heterocycle
In main structure.The heterocycle meaning is for example above-mentioned.
C1-6Alkyl-heterocyclyl groups-C1-3Alkyl refers to by C1-6The C that alkyl-heterocyclyl groups replace1-3Alkyl, wherein passing through C1-3Alkane
Base is connected in the main structure of compound.
C1-6Alkyl-S (O)2Refer to by C1-6Alkyl-substituted S (O)2Group, wherein being connected to compound by S atom
In main structure.
(C1-6Alkyl)2P (O or S)-refer to by 2 C1-6Alkyl-substituted P (O or S) group, wherein being connected by P atom
Onto the main structure of compound.
Term " key " refers to when the atom being connected by key is considered the component part in bigger minor structure, between two
Chemical bond between a atom or two parts.
Terms used herein " pharmaceutically acceptable " refer to when being related to preparation, composition or ingredient to treated
Subject the not lasting adverse effect of general health or do not lose the bioactivity or property of compound, and
Relative non-toxicity.
Terms used herein " bruton's tyrosine kinase " refer to the Bu Ludun junket from homo sapiens (Homo sapiens)
Histidine kinase is disclosed in such as U.S. Patent No. 6326469 (GenBank accession number NP 000052).
Terms used herein " effective quantity " or " therapeutically effective amount " refer to the sufficient amount of the drug or compound given,
Its one or more symptom that will mitigate treated disease or illness to a certain extent.As a result it can be diminution and/or subtract
The change of light sign, symptom or disease reason or any other desired biosystem.For example, for the " effective of therapeutical uses
Amount " be it is required provide so that the clinical symptoms of disease significantly mitigate, without the amount of composition of the excessive toxic side effect of generation, it is described
Composition includes compound disclosed herein." effective quantity " that any individual situation is suitble to can be used and for example incrementally increase agent
The technologies such as quantity research determine.Term " therapeutically effective amount " includes such as prevention effective dose.Compound disclosed herein " effectively
Amount " is effectively to reach desired pharmacological effect or treatment to improve without the amount of excessive toxic side effect.It is understood that
" effective quantity " or " therapeutically effective amount " can be different between subject, and reason is the metabolic, treated of compound
The judgement at person's age, weight, general status, treated disease, the severity of disease being treated and prescribing physician
It is different.Only for example, therapeutically effective amount can be through routine experiment method and determine, include but is not limited to incrementally increase agent
Measure clinical test.
" inhibition ", " inhibition " or " inhibitor " of terms used herein kinases, refers to that phosphate transferase activity is pressed down
System.
Herein, when mentioning " covalently irreversible " inhibition or the similar statement to kinases or kinase activity, compound is indicated
Covalent bond is formed with the amino acid residue on kinases, kinase activity is constantly inhibited.
Autoimmune disease described herein include but is not limited to rheumatoid arthritis, psoriasis arthropathica,
Osteoarthritis, Still disease, adolescent arthritis, lupus, diabetes, myasthenia gravis, Hashimoto thyroiditis, Order first shape
Adenositis, Graves disease, rheumatoid arthritis syndrome, multiple sclerosis, Guillain-Barre syndrome, acute disseminated brain
Myelitis, Addision's disease, opsoclonus-myoclonic syndrome, ankylosing spondylitis, antiphospholipid antibody syndrome, regeneration barrier
Impenetrability anaemia, oneself immunity hepatitis, chylous diarrhea, goodpasture's syndrome, Idiopathic Thrombocytopenic Purpura, optic nerve
Inflammation, chorionitis, primary biliary cirrhosis, Reiter syndrome, takayasu's arteritis, temporal arteritis, warm type autoimmune
Hemolytic anemia, Wegner's granulomatosis, psoriasis, alopecia universalis, behcet disease, confirmed fatigue, familial autonomic nerve
Dysfunction, mullerianosis, interstitial cystitis, neuromyotonia, chorionitis and Vulvodynia.
Heteroimmunity disease described herein includes but is not limited to that graft versus host disease(GVH disease), transplanting, blood transfusion, allergy are anti-
It answers, allergy is (such as to plant pollen, latex, drug, food, insect poison, animal hair, animal scurf, dust mite or cockroach
The allergy of calyx), the hypersensitivity of I type, allergic conjunctivitis, allergic rhinitis and atopic dermatitis.
Inflammatory disease described herein include but is not limited to asthma, inflammatory bowel disease, ecphyaditis, blepharitis, capillary bronchitis,
Bronchitis, bursal synovitis, cervicitis, cholangitis, cholecystitis, colitis, conjunctivitis, cystitis, dacryoadenitis, dermatitis, musculus cutaneus
Inflammation, encephalitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, fascitis, fibrositis, stomach
Inflammation, gastroenteritis, hepatitis, suppurative hidradenitis, laryngitis, mazoitis, meningitis, myelitis myocarditis, myositis, ephritis, oaritis,
Orchitis, osteitis, otitis, pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis, pleurisy, phlebitis, pneumonitis
(pneumonitis), pneumonia (pneumonia), rectitis, prostatitis, pyelonephritis, rhinitis, salpingitis, nasosinusitis,
Stomatitis, synovitis, tendonitis, tonsillitis, uveitis, vaginitis, vasculitis and vulvitis.
Cancer described herein such as B cell proliferative disease includes but is not limited to diffusivity large B cell lymphoid tumor, filter
Bubble property lymthoma, chronic lymphocytic leukemia, chronic lymphocytic leukemia, B cell pre-lymphocytic leukemia, lymph
Plasmacytic lymphoma/macroglobulinemia Waldenstron, splenic marginal zone lymthoma, plasma cell myeloma, plasmacytoma,
Extranodal marginal zone B cell lymphoma, lymphoma nodal marginal zone B cell, lymphoma mantle cell, mediastinum (thymus gland) large B cell
Lymthoma, intravascular large B cell lymphoma, lymphoma primary effusion, Burkitt lymphoma/leukaemia and lymthoma sample meat
The swollen disease of bud.
Thromboembolic disorders described herein include but is not limited to myocardial infarction, (including unstable centering twists angina pectoris
Bitterly), blocking after angioplasty or aortocoronary bypass again or restenosis, apoplexy, transient ischaemic, week
Enclose arterial occlusive disease, pulmonary embolism and Deep vain thrombosis.
It is known in the art to the symptom of each above-mentioned disease, diagnostic assay and prognosis measurement.Referring to example
Such as " Harrison ' s Principles of Internal " the 16th edition, 2004, The McGraw-Hill Companies,
Inc.Dey etc. (2006), Cytojournal 3 (24) and " Revised European American Lymphoma " (REAL)
Categorizing system (see, for example, the operation website of National Cancer Institute (National Cancer Institute)).
Many animal models for establish for treat any aforementioned diseases Btk inhibitor compound (such as covalently not
Reversible Btk inhibitor compound) the range for the treatment of effective dose be useful.
The treatment effect of compound for any aforementioned diseases can be optimised during therapeutic process.For example, by controlling
The subject for the treatment of is amenable to diagnostic assessment, and the alleviation of disease symptoms or pathology and the Btk by giving given dose are pressed down
The inhibition for the internal Btk activity that preparation (such as covalently irreversible Btk inhibitor) obtains is associated.Cell known in the art point
Analysis is determined for the internal work of the Btk in Btk inhibitor (such as covalently irreversible Btk inhibitor) existence or non-existence
Property.For example, the Btk due to activation is phosphorylated in tyrosine 223 (Y223) and tyrosinase 15 51 (Y551), P-Y223 or
The phospho-specif iotac immunocytochemical stain of P-Y551- positive cell can be used for detecting or quantitative determining Bkt in cell mass
Activation situation (such as by facs analysis dye relative to the cell that is unstained).See, for example, Nisitani etc. (1999),
Proc.Natl.Acad.Sci.USA 96:2221-2226.Therefore, the amount for giving the Btk inhibitor compound of subject can
Optimal Btk suppression level is maintained to increase or decrease as needed, for treating the morbid state of subject.
Starting material for synthesizing compound described herein can be synthesized or can obtain from commercial source, such as
But it is not limited to Aldrich Chemical Co. (Milwaukee, Wisconsin), Bachem (Torrance, California)
Or Sigma Chemical Co. (St.Louis, Mo.).Compound described herein to it is other it is related have different substituents
Compound technology well known by persons skilled in the art and Material synthesis can be used, example is as mentioned for example March's
" ADVANCED ORGANIC CHEMISTRY " fourth edition, (Wiley 1992)；" the ADVANCED of Carey and Sundberg
ORGANIC CHEMISTRY " fourth edition, A volumes and B volumes (Plenum 2000,2001)；" the PROTECTIVE of Green and Wuts
GROUPS IN ORGANIC SYNTHESIS " third edition, (Wiley 1999)；" the Reagents for Organic of Fieser
Synthesis " the 1-17 volumes (John Wiley and Sons, 1991)；"Rodd′s Chemistry of Carbon
Compounds " the 1-5 volumes and supplement this (Elsevier Science Publishers, 1989)；"Organic
Reactions " the 1-40 volumes (John Wiley and Sons, 1991)；And " the Comprehensive of Larock
Organic Transformations " (VCH Publishers Inc., 1989) (be fully incorporated herein by reference
In).Other methods for synthesizing compound described herein may refer to international application published WO 01/
No. 01982901；10 (2000) 2167-2170 of Arnold etc., Bioorganic&Medicinal Chemistry Letters；
12 (2002) 1687-1690 of Burchat etc., Bioorganic&Medicinal Chemistry Letters.Preparation is public herein
The conventional method for the compound opened can come from reaction known in the art, and the reaction can be by by those skilled in the art
Reagent and the condition modification that member is deemed appropriate, to be incorporated herein the various parts in the molecule of offer.Synthetic method below
It can be used as guide to utilize.
If desired, routine techniques separation and purifying can be used in reaction product, including but not limited to filters, distills, knot
The methods of brilliant, chromatography.Conventional method characterization, including physical constant and spectrum data can be used in these products.
Synthetic method described herein can be used in compound described herein, and to be prepared as individual isomer or isomers mixed
Close object.
Compound described herein can have one or more stereocenters, and each center may exist R or S
Configuration.Compound provided herein includes the form of all diastereoisomers, enantiomter and epimer, and its is closed
Suitable mixture.If desired, stereoisomer can be obtained by methods known in the art, such as pass through chiral chromatographic column
Separate stereoisomer.
Benefit by known method, such as by chromatography and/or fractional crystallization, can be based on physical chemical differences,
The mixture of diastereoisomer is separated into their individual diastereoisomer.In one embodiment, mapping is different
Structure body can be separated by chiral column chromatography.In other embodiments, can by with optically active compound (example appropriate
Such as alcohol) reaction, the mixture of diastereoisomer is converted by the mixture of enantiomter and separates enantiomter, is separated
It diastereoisomer and converts (such as hydrolysis) out individually diastereoisomer is corresponding pure enantiomter.It is all this
A little isomers are considered as the composition of compositions described herein including diastereoisomer, enantiomter and its mixture
Part.
Method described herein and preparation include using N- oxide, crystal form (it is also assumed that being polymorphic) or
The medicinal acceptable salt and these active metabolisms with the active compound of same type of compound described herein
Object.In some cases, compound can be used as tautomer presence.All tautomers are included in provided herein
In the range of compound.In addition, compound described herein can be present in the form of non-solvent compound and solvate it is medicinal
In acceptable solvent such as water, ethyl alcohol.The solvation form of compound proposed in this paper, which is recognized as, to be disclosed.
It can be by being such as, but not limited to sulphur, sulfur dioxide, triphenylphosphine, lithium borohydride, sodium borohydride, three with reducing agent
Phosphorus chloride, tribromide etc., at 0 DEG C to 80 DEG C and suitable inert organic solvents in, such as, but not limited to acetonitrile, ethyl alcohol,
It is handled in dioxane aqueous solution etc., prepares non-oxidised form from N- oxide.
In some embodiments, pro-drug is made in compound described herein." pro-drug " refers to will be in body
Inside it is converted into the substance of active medicine.Pro-drug be often it is useful because in some cases, they can be than mother
Body drug is easier to give.They can for example be administered orally and be bioavailable, and parent drug then cannot.It is described
Pro-drug also can have the solubility improved than parent drug in Pharmaceutical composition.The example of pro-drug is (unlimited
In) by compound described herein, (" pro-drug ") is given as ester to promote to be conveyed through cell membrane, wherein
Water-soluble is unfavorable for this transfer, but then it is metabolized and is hydrolyzed to carboxylic acid, active entities, once enter intracellular, water
Dissolubility is beneficial.The further example of pro-drug can be the small peptide (polyaminoacid) for being connected to acidic group, wherein peptide
It is metabolized to show active part.In some embodiments, when being administered in vivo, pro-drug is chemically converted as compound
Biology, drug or therapeutic activity form.In some embodiments, pro-drug passes through one or more steps or method quilt
Enzyme is metabolized as the biology, drug or therapeutic activity form of compound.To produce pro-drug, pharmaceutical active compounds are modified,
So that regeneration when the reactive compound is administered in vivo.The pro-drug can be designed as changing the metabolic stability of drug
Or transport features, to cover side effect or toxicity, so as to improve the effect of drug or the other characteristics or property of change drug.Base
In the knowledge to drug effect method and drug metabolism in vivo, once pharmaceutical active compounds be it is known, those skilled in the art are just
The pro-drug of compound can be designed (see, for example, " Nogrady (1985) Medicinal Chemistry A
Biochemical Approach " Oxford University Press, New York, the 388-392 pages；Silverman
(1992) " The Organic Chemistry of Drug Design and Drug Action " Academic Press,
Inc., San Diego, the 352-401 pages, Saulnier etc. (1994), " Bioorganic and Medicinal
Chemistry Letters " volume 4, page 1985).
The prodrug form of compound described herein generates wherein the pro-drug is metabolized in vivo such as preceding institute
Derivative stating, being included within the scope of the claims.In some cases, some compounds as described herein can be it
The pro-drug of its derivative or active compound.
Pro-drug be often it is useful because in some cases, they can be easier compared with parent drug
Ground administration.They can for example be administered orally and be bioavailable, and its parent drug then cannot.In Pharmaceutical composition
In, it also can have improved solubility relative to the parent drug pro-drug.Pro-drug can be designed as reversible
Medicaments derivative is used as modifying agent to enhance drug transport to specific site tissue.In some embodiments, precursor medicine
The design of object increases effective water-soluble.See, for example, Fedorak etc., Am.J.Physiol, 269:G210-218
(1995)；McLoed etc., Gastroenterol, 106:405-413 (1994)；Hochhaus etc., Biomed.Chrom., 6:
283-286(1992)；J.Larsen and H.Bundgaard, Int.J.Pharmaceutics, 37,87 (1987)；
J.Larsen etc., Int.J.Pharmaceutics, 47,103 (1988)；Sinkula etc., J.Pharm.Sd., 64:181-210
(1975)；" Pro-drugs as Novel Delivery Systems " the of T.Higuchi and V.Stella
Volume 14 of A.C.S.Symposium Series；With " the Carriers in Drug Design " of Edward B.Roche
American Pharmaceutical Association and Pergamon Press, 1987, herein by reference by its
In being fully incorporated herein.
Compound described herein includes the compound of isotope labelling, various points with those offers as described herein
Minor and structure are equal, but in fact one or more atoms, by in atom difference atomic weight or mass number and nature
The atomic substitutions being generally found.The example that the isotope of these compounds can be introduced into includes, hydrogen, carbon, nitrogen, oxygen, fluorine and
The isotope of chlorine, such as be respectively2H、3H、13C、14C、15N、18O、17O、35S、18F、36Cl.Certain isotope labellings are retouched herein
The compound stated, such as those radioactive isotopes therein, such as3H and14C is introduced into, and can be used for measuring drug and/or bottom
Object Tissue distribution.Furthermore with isotope such as deuterium, i.e.,2The substitution of H can obtain certain treatments due to the stability of bigger metabolism
Advantage, such as increased Half-life in vivo or reduction need dosage.
In other or further embodiment, by compound described herein give after organism in need
Its metabolism in vivo generates metabolin, and generated metabolin is subsequently used for generating desired effect, including desired therapeutic effect.
Compound described herein can be made into and/or be used as medicinal acceptable salt.Medicinal acceptable salt
Type includes but is not limited to: (1) acid-addition salts, by by the free alkali form of compound and medicinal acceptable inorganic acid reaction
It is formed, described inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid etc.；Or formed with organic acid reaction, it is described
Organic acid for example acetic acid, propionic acid, caproic acid, pentamethylene propionic acid, hydroxyacetic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid,
Maleic acid, fumaric acid, trifluoroacetic acid, tartaric acid, citric acid, benzoic acid, 3- (4- hydroxy benzoyl) benzoic acid, cortex cinnamomi
Acid, mandelic acid, Loprazolam, ethane sulfonic acid, 1,2- ethionic acid, 2- ethylenehydrinsulfonic acid, benzene sulfonic acid, toluenesulfonic acid, 2- naphthalene sulphur
Acid, 4- methyl bicycle-[2.2.2] oct-2-ene -1- formic acid, glucoheptonic acid, 4,4 '-di-2-ethylhexylphosphine oxides-(3- hydroxyl -2- alkene -1- first
Acid), 3- phenylpropionic acid, trimethylace tonitric, butylacetic acid, dodecyl sulphate, gluconic acid, glutamic acid, hydroxynaphthoic acid, bigcatkin willow
Acid, stearic acid, muconic acid etc.；(2) salt, the acid proton in parent compound are formed when being replaced by metal ion, such as
Alkali metal ion (such as lithium, sodium, potassium), alkaline-earth metal ions (such as magnesium or calcium) or aluminium ion；Or it is coordinated with organic base.It can connect
The organic base received includes ethanol amine, diethanol amine, triethanolamine, trimethylamine, N- methyl glucose osamine, etc..Acceptable nothing
Machine alkali includes aluminium hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide etc..
Various method analyses and identification can be used in the corresponding ion balance of medicinal acceptable salt, the method includes
But be not limited to ion-exchange chromatography, ion chromatography, Capillary Electrophoresis, inductively coupled plasma body, atomic absorption spectrum, mass spectrum or
Any combination of them.
Recycle the salt using at least one of following technology: filtering is then filtered, solvent evaporation with non-solvent precipitation,
Or desivac is used in the case where aqueous solution.
It should be understood that the medicinal acceptable salt referred to includes that form or its crystal form, especially solvent is added in solvent
Compound or polymorphic.Solvate includes stoichiometry or non-stoichiometric quantity of solvent, and can be connect with medicinal
It is formed during the solvent the received such as process of water, ethyl alcohol crystallization.Hydrate, or the shape when solvent is alcohol are formed when solvent is water
At alcoholates.In method described herein, the solvate of compound described herein easily can be prepared or be formed.Separately
Outside, compound provided herein can exist with non-solvent compound and solvate forms.In general, solvate forms think with it is non-
Solvate forms are of equal value, the purpose for Compounds and methods for provided herein.
Compound described herein can be various forms, including but not limited to amorphous, spherical and form of nanoparticles.
In addition, compound described herein includes crystal form, also referred to as polymorphic.Polymorphic includes the identical element composition of compound
Different crystal stacked arrangement.Polymorphic usually has different X-ray diffractograms, infrared spectroscopy, fusing point, density, hardness, crystalline substance
Shape, electrical and optical properties, stability and dissolubility.Various factors such as recrystallizes solvent, crystalline rate and storage temperature
It can cause based on single crystal form.
Screening and characterize medicinal acceptable salt, polymorphic and/or solvate can be used multiple technologies completion, described
Technology includes but is not limited to heat analysis, X-ray diffraction, spectrum, steam sorption and microscopy.Heat analysis method focuses on heat
Chemical degradation or thermal physical process comprising but it is not limited to polymorphic transformation, and these methods are for analyzing between polymorphic
Relationship, measurement is weightless, to find glass transition temperature, or is used for excipient Study on Compatibility.These methods include but
It is not limited to differential scanning calorimetry (DSC), modulation differential scanning calorimetry (MDCS), thermogravimetry (TGA) and thermogravimetric
Amount and infrared analysis (TG/IR).Method of X-ray diffraction includes but is not limited to monocrystal and Powder Diffractometer and synchrotron
Source.The various spectral techniques used include but is not limited to Raman, FTIR, UVIS and NMR (liquid and solid state).It is various aobvious
Micromirror technologies include but is not limited to polarization microscope technology, Scanning electron microscopy (SEM) and energy dispersive X-ray point
Analyse (EDX), environmental scanning electron microscope technology and EDX (under gas or water vapour atmosphere), IR microscopy and Raman
(Raman) microscopy.
Carrier herein includes conventional thinner, excipient, filler, adhesive, the wetting agent, disintegration of pharmaceutical field
Agent, sorbefacient, surfactant, absorption carrier, lubricant, it may be necessary to flavouring agent, sweetener etc. be added.The present invention
The diversified forms such as tablet, pulvis, granula, capsule, oral solution and injecting drug use can be made in drug, and the drug of above-mentioned each dosage form is equal
It can be prepared according to the conventional method of pharmaceutical field.
IC used herein50Refer to, amount, concentration or the dosage of fc-specific test FC compound, in the inhibition of measurement such as Btk
In the test of such reaction, reach ceiling effect 50% inhibits.
Among this entire specification, group and its substituent group can be selected by those skilled in the art, to provide
Stable compound.
Embodiment
Non-limiting embodiment in detail below is to be interpreted as being merely illustrative, not limit in any way originally
Invention.Although without being described in further detail, it is believed that those skilled in the art can be based on description herein, completely benefit
With the present invention.
The synthesis of compound
In synthetic schemes below, following abbreviation is used:
Boc: tert-butoxycarbonyl；
DPB: connection boric acid pinacol ester；
Et: ethyl；
Rt/RT: room temperature；
Me: methyl；
DIEA:N, N- diisopropylethylamine；
DCM: methylene chloride；
NMP:N- methyl pyrrolidone；
TEA: triethylamine；
TFA: trifluoroacetic acid；
THF: tetrahydrofuran；
HATU:2- (7- aoxidizes benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester；
DMF: dimethylformamide.
Synthetic schemes I
Step 1:
By guanidine carbonate (281g, 1.56mol), the mixed solution of DIEA (540mL, 3.12mol) and NMP (1L) are in stirring shape
It is heated to 150-160 DEG C under state, the nmp solution (100ml) of the bromo- 2- fluorobenzaldehyde (250g, 1.20mol) of 5- is slowly added dropwise.Instead
After answering 3 hours, 100 DEG C are cooled to, ice (2kg) and water (4L) is added, there is Tan solid precipitation, continues stirring 30 minutes, subtracts
Press filtration is washed with water (1L) and ethyl alcohol (1L), and gained filter cake is transferred in 5L flask, ethyl alcohol (2L) stirring 2 hours is added, so
After filter, successively with ethyl alcohol (0.5L), toluene/ethanol (1:1,0.5L) and toluene (0.5L) washing, it is dry after compound 3,
For light yellow solid (168g, 48%).
Step 2:
By compound 3 (5.0g, 22.3mmol), cuprous iodide (4.30g, 22.3mmol) and diiodomethane (9.0mL,
It 114mmol) is dissolved in THF (100mL), is added isoamyl nitrite (9.0mL, 68mmol), is heated to reflux 2.5 under nitrogen atmosphere
Hour.Reaction solution is cooled to room temperature, and ethyl acetate (500mL) and 1N HCl (500mL) is added, water phase is after liquid separation with ethyl acetate
It is extracted twice, merges organic phase, washed with saturated aqueous ammonium chloride, anhydrous magnesium sulfate dries, filters concentration, gained grease
Compound 4 (2.60g, 35%) is obtained through column chromatography for separation (methylene chloride 100%), is near-white powder solid.
Step 3:
Compound 4 (4.0g, 12.0mmol) and N-Boc- m-phenylene diamine (MPD) (3.7g, 17.9mmol) are dissolved in Isosorbide-5-Nitrae-dioxy six
In ring (200mL), be added bis- (diphenylphosphine phenyl ether) palladium chlorides (II) (820mg, 1.1mmol) and cesium carbonate (19.5g,
60.0mmol), which is replaced three times with argon gas, is heated to 100 DEG C under an argon and is reacted 20 hours, and TLC detection is former
Material is exhausted.Reaction solution is filtered under diminished pressure after being cooled to room temperature through diatomite, and gained filtrate is diluted with ethyl acetate and water, liquid separation
Water phase is extracted twice afterwards with ethyl acetate, merges organic phase, and with the washing of saturated common salt aqueous solution, anhydrous magnesium sulfate is dried, filtered
Concentration, (n-hexane: ethyl acetate=2:1 to 1:1) obtains compound 5 (3.4g, 68%) to column chromatography for separation, is buff powder
Solid.
Step 4:
Compound 5 (3.37g, 8.1mmol) and connection boric acid pinacol ester (12.17g, 3.1mmol) are dissolved in dimethyl methyl
In amide (200mL), be added bis- (diphenylphosphine phenyl ether) palladium chlorides (II) (594mg, 0.8mmol) and potassium acetate (4.0g,
40.5mmol), which is replaced three times with argon gas, is heated to 80 DEG C under an argon and is reacted 20 hours, TLC detects raw material
It is exhausted.Reaction solution is cooled to room temperature, and is filtered through diatomite, and solvent is removed in the rotation of gained filtrate decompression, dilute with ethyl acetate and water
It releases, water phase is extracted twice after liquid separation with ethyl acetate, merges organic phase, and with the washing of saturated common salt aqueous solution, anhydrous magnesium sulfate is dry
Dry, filtering and concentrating, (n-hexane: ethyl acetate=10:1 to 5:1) obtains compound 6 (2.63g, 70%) to column chromatography for separation, is
Buff powder solid.
Step 5:
The iodo- 4- methyl benzoic acid (3.2g, 12.2mmol) of 3- is scattered in 50mL thionyl chloride, which is warming up to
82 DEG C are kept stirring reflux 1 hour, are cooled to room temperature.Reaction solution is after keeping being slowly stirred lower addition 50mL toluene, decompression rotation
Inspissation is reduced to light yellow oil.After the concentrate is dissolved with 100mL methylene chloride, 3- trifluoromethylbenzene is added in this solution
Amine (2.3mL, 18.3mmol) and diisopropylethylamine (9mL), the reaction system are precipitated a large amount of white after being stirred at room temperature overnight
Color solid.Reaction solution is scattered in ethyl acetate after being concentrated under reduced pressure, and successively uses saturated ammonium chloride solution and saturated common salt water washing,
Final organic phase is dry with anhydrous magnesium sulfate, is concentrated under reduced pressure, purify to obtain compound 8 (3.5g, 71%) with silica gel column chromatography be
White solid.
Step 6:
Compound 6 (690mg, 1.49mmol) and compound 8 (910mg, 2.25mmol) are dissolved in the mixing of acetonitrile and water
In solution in (90mL/30mL), bis- (diphenylphosphine phenyl ether) palladium chlorides (II) (150mg, 0.15mmol) and carbonic acid is added
Potassium (1.25g, 9.00mmol), the reaction system are replaced three times with argon gas, are heated to 82 DEG C under an argon and are reacted 24 hours,
TLC detection consumption of raw materials finishes.Reaction solution is cooled to room temperature, and is filtered through diatomite, and solvent is removed in the rotation of gained filtrate decompression, with acetic acid
Ethyl ester and water dilution, water phase is extracted twice after liquid separation with ethyl acetate, merges organic phase, with the washing of saturated common salt aqueous solution, nothing
Water magnesium sulfate dries, filters concentration, column chromatography for separation (n-hexane: ethyl acetate=5:1 to 2:1) obtain compound 9 (650mg,
It 71%), is buff powder solid.
Step 7:
It disperses compound 9 (650mg, 1.06mmol) in 100mL methylene chloride, is being kept stirring lower 15mL trifluoro second
In the instillation reaction system of acid slowly.After final reaction system is kept stirring 1 hour at room temperature, it is concentrated under reduced pressure to give solid fraction
Object.Saturated sodium bicarbonate solution and saturated common salt water washing are successively used after residue with Ethyl acetate dissolution.Final organic phase
With anhydrous magnesium sulfate it is dry after be concentrated under reduced pressure, concentrate silica gel column chromatography purify (n-hexane: ethyl acetate=1: 1~1: 2~
1: 4) obtaining compound 10 (501mg, 92%), be near-white solid.
Synthetic schemes II
Step 1:
Disperse compound 10 (501mg, 0.98mmol) in the in the mixed solvent (50mL, 4: 1V/V) of THF and water, then
It is added diisopropylethylamine (0.23mL, 1.17mmol).In the case where keeping being slowly stirred, acryloyl chloride (96 μ L, 1.17mmol) is slow
In slow instillation reaction system.It after 2 hours are stirred at room temperature in reaction solution, is concentrated under reduced pressure, residue with Ethyl acetate dissolution
The successively citric acid solution and saturated common salt water washing with 10% afterwards.Final organic phase is dense to depressurize after anhydrous magnesium sulfate drying
Contracting, concentrate silica gel column chromatography purify (methylene chloride: methanol=20: 1) obtain compound 11 (400mg, yield: 72%) being close
White powdery solids.
Synthetic schemes III
Step 1:
The chloro- 4- of 3- (4- piperidines -1- methyl) benzoate hydrochlorate (1.7g, 5.6mmol) is scattered in 100mL thionyl chloride
In, which is warming up to 82 DEG C and is kept stirring reflux 24 hours, is cooled to room temperature.Reaction solution is keeping being slowly stirred lower add
After entering 50mL toluene, vacuum rotary steam is concentrated to give near-white solid.The solid with 100mL isopropanol dissolve after, in this solution plus
Enter the iodo- 4- methylaniline (1.8g, 7.7mmol) of 3- and potassium carbonate (3.8g, 27.5mmol), which is warming up to 82 DEG C of guarantors
It holds and is stirred at reflux 24 hours, TLC detection consumption of raw materials finishes.Solvent is removed in decompression rotation, molten with ethyl acetate and saturated sodium bicarbonate
Liquid dilution, water phase merges organic phase with ethyl acetate extraction four times after liquid separation, with the washing of saturated common salt aqueous solution, anhydrous slufuric acid
Magnesium dries, filters concentration, column chromatography for separation (methylene chloride: methanol=10:1 to 5:1) obtains compound 13 (700mg, 26%),
For greenish yellow solid.
Step 2:
Compound 6 (1.1mg, 2.38mmol) and compound 13 (700mg, 1.45mmol) are dissolved in the mixing of acetonitrile and water
In solution in (90mL/30mL), bis- (diphenylphosphine phenyl ether) palladium chlorides (II) (150mg, 0.15mmol) and carbonic acid is added
Potassium (1.3g, 9.00mmol), the reaction system are replaced three times with argon gas, are heated to 82 DEG C under an argon and are reacted 24 hours, TLC
Detection consumption of raw materials finishes.Reaction solution is cooled to room temperature, and is filtered through diatomite, and solvent is removed in the rotation of gained filtrate decompression, with acetic acid second
Ester and water dilution, water phase merges organic phase with ethyl acetate extraction four times after liquid separation, anhydrous with the washing of saturated common salt aqueous solution
Magnesium sulfate dries, filters concentration, column chromatography for separation (methylene chloride: methanol=20:1 to 10:1) obtain compound 14 (551mg,
It 55%), is dark gray powder solid.
Step 3:
It disperses compound 14 (551mg, 0.80mmol) in 240mL methylene chloride, is being kept stirring lower 60mL trifluoro
In the instillation reaction system of acetic acid slowly.After final reaction system is kept stirring 3 hours at room temperature, it is concentrated under reduced pressure to give solid
Shape object.Saturated sodium bicarbonate solution and saturated common salt water washing are successively used after residue with Ethyl acetate dissolution.It is final organic
Mutually with anhydrous magnesium sulfate it is dry after be concentrated under reduced pressure, concentrate silica gel column chromatography purify (methylene chloride: methanol=20:1 to 10:
1) compound 15 (451mg, 84%) is obtained, is yellow greenish powder solid.
Step 4:
By compound 15 (451mg, 0.76mmol) be scattered in THF and methylene chloride in the mixed solvent (300mL, 1: 1V/
V), triethylamine (0.3mL, 1.53mmol) then is added.In the case where keeping being slowly stirred, acrylic anhydride (46 μ L, 0.40mmol) is slow
In slow instillation reaction system.It after 12 hours are stirred at room temperature in reaction solution, is concentrated under reduced pressure, residue with Ethyl acetate dissolution
Saturated sodium carbonate solution and saturated common salt water washing are successively used afterwards.Final organic phase is concentrated under reduced pressure after being dried with anhydrous magnesium sulfate,
Concentrate silica gel column chromatography purify (methylene chloride: methanol=20:1) compound 16 (411mg, yield: 84%), to be pale yellow
Color pulverulent solids.
Synthetic schemes IV
Step 1:
The bromo- niacin of 5- (3.2g, 2.5mmol) is scattered in 20mL thionyl chloride, which is warming up to 82 DEG C of holdings
It is stirred at reflux 2 hours, is cooled to room temperature.After keeping being slowly stirred lower addition 20mL toluene, vacuum rotary steam is concentrated into reaction solution
Light yellow oil.After the concentrate is dissolved with 100mL methylene chloride, the iodo- 4- methylaniline of 3- is added in this solution
(874mg, 3.8mmol) and triethylamine (0.5mL), which, which is stirred at room temperature overnight to raw material to detect, disappears.Reaction
Liquid is scattered in ethyl acetate after being concentrated under reduced pressure, and successively uses 2N HCl solution and saturated common salt water washing, final organic phase nothing
Water magnesium sulfate is dry, is concentrated under reduced pressure, obtains compound 18 (1.0g, 96%), is light yellow solid.
Step 2:
Compound 6 (345mg, 0.75mmol) and compound 18 (470mg, 1.13mmol) are dissolved in the mixing of acetonitrile and water
In solution in (90mL/30mL), bis- (diphenylphosphine phenyl ether) palladium chlorides (II) (55mg, 0.08mmol) and potassium carbonate is added
(621mg, 4.50mmol), the reaction system are replaced three times with argon gas, are heated to 82 DEG C under an argon and are reacted 24 hours, TLC
Detection consumption of raw materials finishes.Reaction solution is cooled to room temperature, and is filtered through diatomite, and solvent is removed in the rotation of gained filtrate decompression, with acetic acid second
Ester and water dilution, water phase is extracted twice after liquid separation with ethyl acetate, merges organic phase, anhydrous with the washing of saturated common salt aqueous solution
Magnesium sulfate dries, filters concentration, column chromatography for separation (n-hexane: ethyl acetate=5:1 to 3:2) obtain compound 19 (391mg,
It 63%), is buff powder solid.
Step 3:
Compound 19 (125mg, 0.20mmol) and dimethyl phosphine (24mg, 0.30mmol) are dissolved in dimethyl formyl
In amine (15mL), be added bis- (diphenylphosphine phenyl ether) palladium chlorides (II) (15mg, 0.02mmol) and potassium acetate (98mg,
1.0mmol), which is replaced three times with argon gas, is heated to 100 DEG C under an argon and is reacted 24 hours, TLC detects raw material
It is exhausted.Reaction solution is cooled to room temperature, and is filtered through diatomite, and solvent is removed in the rotation of gained filtrate decompression, with ethyl acetate and saturation
Sodium bicarbonate solution dilution, water phase is extracted twice after liquid separation with ethyl acetate, is merged organic phase, is washed with saturated common salt aqueous solution
Wash, anhydrous magnesium sulfate dries, filters concentration, column chromatography for separation (methylene chloride: methanol=20: 1) obtain compound 20 (80mg,
It 64%), is dark brown powder solid.
Step 4:
It disperses compound 20 (80mg, 0.13mmol) in 15mL methylene chloride, is being kept stirring lower 4mL trifluoroacetic acid
In instillation reaction system slowly.After final reaction system is kept stirring 2 hours at room temperature, it is concentrated under reduced pressure to give solid fraction
Object.Saturated sodium bicarbonate solution and saturated common salt water washing are successively used after residue with Ethyl acetate dissolution.Final organic phase
With anhydrous magnesium sulfate it is dry after be concentrated under reduced pressure, concentrate, which is purified with silica gel column chromatography, (methylene chloride: methanol=20: 1) to be changed
It closes object 21 (46mg, 69%), is yellow solid.
Step 5:
Disperse compound 21 (30mg, 0.06mmol) in the in the mixed solvent (15mL, 4: 1V/V) of THF and water, then
It is added triethylamine (12 μ L, 0.09mmol).In the case where keeping being slowly stirred, acryloyl chloride (7 μ L, 0.09mmol) slowly instills anti-
It answers in system.It after 12 hours are stirred at room temperature in reaction solution, is concentrated under reduced pressure, is successively used after residue with Ethyl acetate dissolution
10% citric acid solution and saturated common salt water washing.Final organic phase after anhydrous magnesium sulfate drying to be concentrated under reduced pressure, concentrate
Purify that (methylene chloride: methanol=15: 1) obtaining compound 22, (20mg, yield: 60%), being chartreuse with silica gel column chromatography
Pulverulent solids.
The external inhibitory activity of test example 1:Btk is analyzed
In cell-free kinase assays, using method as described below or the like measurement the compounds of this invention to Btk
503nhibiting concentration (IC50)。
Use time-resolved fluorescence Resonance energy transfer (time-resolved fluorescence resonance
Energy transfer) (TR-FRET) method measurement Btk kinase activity.Using 96 hole assay plates, in 50 μ L reaction volumes
It is measured.By kinases, inhibitor, ATP (in the K of kinasesm) and 1 μM of peptide substrates (biotin-AVLESEEELYSSARQ-NH2)
It is incubated for 1 hour in reaction buffer (pH 7.4), the reaction buffer is by 20mM Tris, 50mM NaCl, MgCl2(5-
25mM, depend on kinases), MnCl2(0-10mM), 1mM DTT, 0.1mM EDTA, 0.01% bovine serum albumin(BSA), 0.005%
Tween-20 and 10%DMSO composition.Pass through 1.2 equivalents in the 1x Lance buffer (Perkin-Elmer) of 25 μ L of addition
EDTA (relative to bivalent cation) quenching reaction object.Streptavidin-APC (Perkin-Elmer) is added in 25 μ L volumes
With Eu label p-Tyr100 antibody (Perkin-Elmer) 1x Lance buffer, respectively obtain final concentration of 100nM and
2.5nM incubates the mixture 1 hour.Using multi-mode plate reader (multimode plate reader), 330nm's
Excitation wavelength (λEx) and 615nm and 665nm Detection wavelength (λEm) under measure TR-FRET signal.By under 665nm and 615nm
Fluorescence than determine activity.To each compound, the enzymatic activity under various concentration compound is determined.Negative control reaction exists
Lack and carry out in the case of inhibitor (do six duplicate), and determines baseline fluorescence level without enzyme control with two.Make
With program Batch Ki(Kuzmic etc. (2000), Anal.Biochem.286:45-50) fitting obtains IC50。
According to above-mentioned synthetic schemes I, II, III and IV, the embodiment of the present invention compound 1~41 is synthesized.Embodiment chemical combination
Object is characterized as below shown in table 1.In the external inhibitory activity analysis of Btk, the embodiment of the present invention compound 1~41 is determined
IC50Value.According to IC50Section locating for value provides IC50Value, wherein " +++ " represents IC50<100nM；" ++ " represents 100nM < IC50
<1000nM；"+" represents 1000nM < IC50<10000nM。
The synthesis of 1 embodiment compound of table and Btk IC50Value
For the efficacy data that last is arranged by above-mentioned table 1 as it can be seen that being directed to Btk, the compound of the application can obtain IC50It is less than
The compound inhibitory effect of the excellent effect of 100nM, especially embodiment 2,12,14,40,41 is especially excellent.
Test example 2: the covalent irreversible inhibitory activity (inhibiting rate test) of compound
It uses homogeneous phase time discrimination fluorescence technology HTRF (Homogeneous Time-Resolved Fluorescence)
Method detects kinase activity, 20 μ L of end reaction volume.Reaction buffer is HTRF tyrosine-kinase enzyme reagent kit (Cisbio, goods
Number: 1 × kinase buffer++1mM DTT+5mM MgCl 62TK0PEC)2+50nM Supplement Enzymatic
Buffer takes each compound or 1% dimethyl sulfoxide (as control) mixing of concentration shown in 0.5ngBtk kinases and following table, presses
Shown in table, for each sample respectively be incubated for 0 minute, 5 minutes, 10 minutes, 15 minutes, 30 minutes after, 1 μM of kit institute is added
Substrate and 20 μM of atriphos are provided, according to the step recorded in HTRF tyrosine kinase kit specification after reaction 1 hour
The rapid detection reagent that is added terminates, and microplate reader (Perkin Elmer Envision) reads data.1% dimethyl sulfoxide is added
Sample is as control, with its kinase activity under same incubation time for 100%, to living through each compound treated kinases
Property is normalized, and calculates the kinase inhibition rate of each compound, as shown in table 2 below.
2 bruton's tyrosine kinase maximum inhibition of table
Experiments have shown that in-N (R in the application formula (I)1-L1)(R2-L2) compound in part comprising alkenyl is (for example, table
The compound of 1 embodiment 2 and 12) to the active inhibiting rate of bruton's tyrosine kinase as compound and bruton's tyrosine swash
The cultivation time of enzyme extends and dramatically increases, and embodies covalent Irreversible binding activity；And control compounds (1 embodiment 11 of table
Compound) inhibiting rate amplitude of variation it is significantly smaller.
Test example 3: the covalent irreversible inhibitory activity of compound (fluorescence probe combines test)
Reaction buffer is the 1 × kinase of HTRF tyrosine-kinase enzyme reagent kit (Cisbio, article No.: 62TK0PEC)
buffer+1mM DTT+5mM MgCl2+ 50nM Supplement Enzymatic buffer, takes 40ng Btk kinases and 1%
The compound of 1 embodiment 2 of dimethyl sulfoxide or the table of 1 μM of concentration, the compound of 1 embodiment 12 of table mix respectively, are incubated for 10 minutes
Afterwards, 0.5 μM of reported covalent fluorescence probe [Scientific Reports, 2015,5,16136] is added, reacts 30 minutes,
2 × loading buffer that equivalent responses volume is added terminates reaction.20 μ L of end reaction volume.10 μ L loadings are respectively taken,
Fluorescent scanning is carried out after SDS-PAGE electrophoresis.As the result is shown: the Btk band of 1%DMSO processing shows strong fluorescence, and table 1 is implemented
Basic unstressed configuration band occurs in the swimming lane of the compound of example 2 and 12.Experiments have shown that: in-N (R in the application formula (I)1-L1)
(R2-L2) compound (for example, compound of 1 embodiment 2,12 of table) in part comprising alkenyl can effectively prevent covalently can not
Inverse combination of the fluorescence probe for bruton's tyrosine kinase.
It is understood that embodiment described herein the purposes for being only used for illustrating with embodiment, and accordingly
Made various modifications or change can make enlightenment to those skilled in the art, and the spirit including applying herein and
In the range of range and appended claims.Herein cited all publications, patents and patent applications all pass through reference
All it is incorporated by herein with for all purposes.
Claims (14)
1. a kind of compound or its pharmaceutically acceptable salt of formula (I):
Wherein,
W is selected from H, C1-6Alkyl ,-(NH-CO)n-L-L3,-(CO-NH)n-L-L3,
Wherein,
L is key, C1-3Alkylidene or C2-3Alkenylene,
L3For the C optionally replaced by 1,2 or 3 substituent group selected from the following3-8Naphthenic base, aryl, bicyclic heteroaryl are thick miscellaneous
Ring, oxo condensed hetero ring: halogen, amino, C1-6Alkyl, C1-6Alkoxy, halogenated C1-6Alkyl, heterocycle, C1-6Alkyl-heterocyclyl groups,
C1-6Alkyl-heterocyclyl groups-C1-3Alkyl, C1-6Alkyl-S (O)2, (C1-6Alkyl)2P (O or S)-,
N is 0 or 1,
X is selected from H, halogen and C1-6Alkyl,
R1And R2Independently of one another, in the same manner or differently, H, C (O) and S (O) are selected from2；
L1And L2Independently of one another, in the same manner or differently, selected from for optionally by C1-3Alkyl-substituted C2-3Alkenyl；
On condition that working as R1When for H, L1It is not present, and works as R2When for H, L2It is not present.
2. compound as described in claim 1 or its pharmaceutically acceptable salt, wherein
W is selected from-(NH-CO)n-L-L3,-(CO-NH)n-L-L3,
Wherein,
L is key,
L3To be optionally selected from F, Cl, Br, amino, methyl, methoxyl group, ethyoxyl, propoxyl group, CF by 1,2 or 33,CH3-S(O)2, (CH3)2The cyclopropyl that the substituent group of P (O or S)-replaces, phenyl, naphthalene, furans
Base, thienyl, pyrrole radicals, thiazolyl, imidazole radicals, pyrazolyl, pyridyl group, pyrimidine radicals, pyrazinyl, pyridazinyl, benzheterocycle base,
Pyrazolo heterocycle, oxo benzheterocycle base,
N is 1.
3. compound as claimed in claim 2 or its pharmaceutically acceptable salt, wherein the benzheterocycle base is benzo
Nitrogen heterocycle, the pyrazolo heterocycle are pyrazolo nitrogen heterocycle, and the oxo benzheterocycle base is that oxo benzo contains
Azacyclyl.
4. compound as claimed in claim 3 or its pharmaceutically acceptable salt, wherein the benzheterocycle base are as follows:The pyrazolo heterocycle isThe oxo
Benzheterocycle base is
5. compound as described in any one of claims 1 to 4 or its pharmaceutically acceptable salt, wherein R1And R2In
One is H, and another one is C (O).
6. such as compound according to any one of claims 1 to 5 or its pharmaceutically acceptable salt, wherein X is selected from H, F,
Cl and methyl, the-N (R1-L1)(R2-L2) it is-NH2Or-NH-C (O)-CH=CH2。
7. such as compound according to any one of claims 1 to 6 or its pharmaceutically acceptable salt, wherein L3For optionally
F, Cl, Br, methyl, CF are selected from by 1,2 or 33,CH3-S(O)2, (CH3)2The substituent group of P (O)-
Substituted phenyl, 3- pyridyl group, 4- pyridyl group,
8. a kind of compound, is selected from:
9. compound as described in claim 8, is further selected from:
10. such as compound according to any one of claims 1 to 9, for inhibiting bruton's tyrosine kinase active.
11. compound as claimed in claim 10, the compound can inhibit bruton's tyrosine in a manner of covalent bond
Kinase activity.
12. the compound as described in any one of claim 1~11, the compound can on bruton's tyrosine kinase
Cysteine residues formed covalent bond.
13. a kind of Pharmaceutical composition, it includes the compounds and medicine described in any one of claim 1~12 of therapeutically effective amount
Acceptable carrier or excipient on.
14. compound described in any one of claim 1~12 or its pharmaceutically acceptable salt preparation for treat with
Purposes in the drug of lower disease or situation: autoimmune disease, heteroimmunity disease, inflammation disease, cancer, thrombus bolt
Fill in disease.
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