CN109422696A - The inhibitor of bruton's tyrosine kinase - Google Patents
The inhibitor of bruton's tyrosine kinase Download PDFInfo
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- CN109422696A CN109422696A CN201710787857.6A CN201710787857A CN109422696A CN 109422696 A CN109422696 A CN 109422696A CN 201710787857 A CN201710787857 A CN 201710787857A CN 109422696 A CN109422696 A CN 109422696A
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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
- C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
- C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
- C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
- C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
- C07D239/48—Two nitrogen atoms
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
Abstract
The invention discloses a kind of logical formula (I) compounds that can inhibit Btk, 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.Present invention offer can inhibit the active compound of bruton's tyrosine kinase in a manner of covalent bond, for example in a manner of covalent reversible.
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 inhibit the active compound of bruton's tyrosine kinase in a manner of covalent reversible.The compound of the present invention can be used for
Treat 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.
In addition, tyrosine kinase inhibitor can be divided into according to the difference of inhibitor and the combination of tyrosine kinase
Reversible and irreversible.Current small molecule tyrosine kinase inhibitors belong to reversible inhibitor more, and usually these are reversible
The combination of the binding domain of inhibitor and tyrosine kinase be by weaker, reversible active force, such as hydrogen bond, Van der Waals force and dredge
Water effect etc. is realized.This weaker and reversible active force there are poor selectivity, drug effect it is not strong enough and persistently and
The disadvantages of easily causing drug resistance.On the other hand, irreversibility tyrosine kinase inhibitor is with ATP on covalent bond and tyrosine kinase
Binding domain is combined, so that target spot be made permanently to inactivate.Due to its unique mechanism of action, the suppression of irreversibility tyrosine kinase
Preparation can efficiently solve the disadvantages mentioned above of invertibity tyrosine kinase inhibitor.But it is at the same time, this kind of based on irreversible
The inhibitor of Covalent bonding together also bring it is more due to " effect of missing the target " caused by toxic side effect worry (referring to document
3)。
Some in present inventor report a kind of (aminophenyl for Btk in patent application before this
Amino) pyrimidine radicals benzamides inhibitor (referring to document 4), belong to irreversible covalency inhibitor.
For Btk reversible covalent inhibit drug so far there is no.If the reversible covalent suppression for Btk can be developed
Preparation, then the advantages of may taking into account above two inhibitor, have high-drug-effect, are not easy drug resistance and the spy of hypotoxicity simultaneously
Point.
Existing technical literature
Document 1:CN101610676A；
Document 2:Zhengying Pan etc., ChemMedChem 2007,2,58-61；
Document 3: Guo Jianjun etc., Chinese Pharmacological Bulletin, 2015,31 (6), 749-754；
Document 4:WO2013060098A1
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 chemical combination for being directed to bruton's tyrosine kinase (BTK)
Object.Further, the present invention also provides in a manner of covalent bond, especially covalent reversible mode inhibits kinases (especially cloth Shandong
Tyrosine kinase) active compound.
Means for solving the problems
In one aspect of the invention, provide a kind of formula (I) compound or its pharmaceutically can or its pharmaceutically may be used
The salt of receiving:
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 heteroaryl is such as
The C3-8Naphthenic base, aryl or heteroaryl are optionally replaced by following substituent groups: halogen, amino, C1-6Alkyl,
C1-6Alkoxy, halogenated C1-6Alkyl, C1-6Alkyl-heterocyclyl groups-C1-3Alkyl,
N is 0 or 1,
X is selected from H, halogen such as F and Cl or C1-6Alkyl such as methyl,
R is amino, the C optionally replaced by 1,2 or 3 substituent group selected from the following1-6Alkyl, C3-8Naphthenic base, aryl
Or heteroaryl: halogen, hydroxyl, amino, C1-6Alkyl, C1-6Alkyl amino, C1-6Alkoxy, C2-6Alkynyl-C1-3Alkyl-(CO-
NH)-。
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, methoxyl group, ethyoxyl, propoxyl group, CF by 1,2 or 33OrSubstituent group replace cyclopropyl, phenyl, naphthalene,
N is 1.
In another preferred embodiment, X is selected from H, F, Cl or methyl.
In another preferred embodiment, R is optionally to be selected from F, Cl, Br, hydroxyl, amino, first by 1,2 or 3
Amino, the ring of the substituent group substitution of base, ethyl, methylamino, ethylamino, methoxyl group, ethyoxyl, acetenyl-propyl-(CO-NH)-
Propyl, isopropyl, tert-butyl, phenyl, furyl, imidazole radicals, thienyl, pyrrole radicals, pyridyl group.
In another preferred embodiment, R is optionally to be selected from F, hydroxyl, amino, methyl, first ammonia by 1,2 or 3
Base, the amino of the substituent group substitution of acetenyl-propyl-(CO-NH)-, cyclopropyl, isopropyl, tert-butyl, phenyl, furyl, miaow
Oxazolyl, thienyl.In another aspect of the present invention, a kind of compound is provided, is selected from:
In another aspect of the present invention, the compounds of this invention is for inhibiting bruton's tyrosine kinase active.
In another aspect of the present invention, the compounds of this invention can inhibit bruton's tyrosine in a manner of covalent bond
Kinase activity.
In another aspect of the present invention, the compounds of this invention can be residual with the amino acid on bruton's tyrosine kinase
Base forms reversible covalent bonds.
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 compound of the present invention can inhibit bruton's tyrosine kinase active in a manner of covalent bond.Further, originally
Invention is additionally provided inhibits the active compound of bruton's tyrosine kinase in a manner of covalent reversible.
Detailed description of the invention
[Fig. 1] indicates bruton's tyrosine kinase maximum inhibition.
[Fig. 2] indicates that DMSO, compound R PG41 and Yi Lu detect figure for the Western blot of Buddhist nun.Wherein, " 5min ",
" 10min " indicate fluorescence probe with dilute after the incubation of kinases compound corresponding time when the case where；Band corresponding to " BTK "
The BTK content in each loading hole when carrying out Western Blot detection with BTK after SDS-PAGE is shown；1X, 3X, 5X and 10X are indicated
Diluted extension rate is carried out after by kinases and compound incubation 15min.
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.
The example of " heteroaryl " includes:
Deng.
“C3-8Naphthenic base " refer to nonaromatic, only containing carbon and hydrogen and annular atom number be 3-8 carbon atom
One or more cyclic groups, and it can be saturation, part not
It is saturation or complete unsaturated.C3-8The example of naphthenic base includes following part:
Deng.
" 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.
C1-6Alkyl amino refers to through C1-6Substituted amino, substituent group can be 1 or 2, such as CH3- NH-, CH3-
CH2- NH-, (CH3)2N-, wherein being connected in the main structure of compound by amino.
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.The heterocycle refer to containing at least one be selected from containing nitrogen-atoms, oxygen atom and/or
Heteroatomic monocycle, the ring system of two rings or loop coil of sulphur atom.Heterocycle can be 3,4,5,6,7 or 8 member rings.The heterocycle of monocycle
The example of base includes but is not limited to azetidinyl, nitrogen heterocyclic heptyl, aziridine base, Diazesuberane, 1,3-
Diaza-cyclohexane, 1,4- diaza-cyclohexane etc..
C2-6Alkynyl-C1-3Alkyl-(CO-NH)-refer to by C2-6Alkynyl-C1-3Alkyl-substituted (CO-NH)-, wherein passing through
(CO-NH)-be connected in the main structure of compound.
Term " alkynyl " refers to the linear chain or branched chain hydrocarbyl group containing 2-10 carbon atom He at least one tri- key of C-C.Alkynyl
Representative example include but is not limited to acetenyl, 1- propinyl, 2-propynyl, 1,1- dimethyl Propargyl etc..
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, mention and " covalent reversible " of kinases or kinase activity being inhibited or when similar statement, indicate compound with
Amino acid residue on kinases with can inverse form form covalent bond, to inhibit kinase activity.
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 are for establishing controlling for the covalent reversible Btk inhibitor compound for treating any aforementioned diseases
It is useful for treating the range of effective dose.
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, by the alleviation of disease symptoms or pathology with can by giving the covalent of given dose
The inhibition for the internal Btk activity that inverse Btk inhibitor obtains is associated.Cell analysis known in the art is determined for can
The activity in vivo of Btk when inverse Btk inhibitor existence or non-existence.For example, since the Btk of activation is at tyrosine 223 (Y223)
It is phosphorylated with tyrosinase 15 51 (Y551), therefore the phospho-specif iotac immunocytochemistry of P-Y223 or P-Y551- positive cell
Dyeing can be used for detect or quantitative determine cell mass in Bkt activation situation (such as by facs analysis dye relative to
Be unstained cell).See, for example, Nisitani etc. (1999), Proc.Natl.Acad.Sci.USA 96:2221-2226.Cause
This, the amount for giving the Btk inhibitor compound of subject, which can according to need, to be increased or decreased to maintain optimal Btk to inhibit
Level, 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 incorporated by 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 in vivo
It is converted into the substance of active medicine.Pro-drug be often it is useful because in some cases, they can be than parent medicine
Object is easier to give.They can for example be administered orally and be bioavailable, and parent drug then cannot.The precursor
Drug also can have the solubility improved than parent drug in Pharmaceutical composition.The example of pro-drug is that (being not limited to) will
Compound described herein gives (" pro-drug ") as ester to promote to be conveyed through cell membrane, wherein water-soluble
Solution property is unfavorable for this transfer, but then it is metabolized and is hydrolyzed to carboxylic acid, active entities, once enter intracellular, water dissolution
Property is beneficial.The further example of pro-drug can be the small peptide (polyaminoacid) for being connected to acidic group, and wherein peptide is by generation
It thanks to show active part.In some embodiments, when being administered in vivo, life that pro-drug is chemically converted as compound
Object, drug or therapeutic activity form.In some embodiments, pro-drug is by one or more steps or method by enzyme generation
Thank the biology for compound, drug or therapeutic activity form.To produce pro-drug, pharmaceutical active compounds are modified, so that
The regeneration when reactive compound is administered in vivo.The pro-drug can be designed as changing the metabolic stability of drug or turn
Characteristic is transported, to cover side effect or toxicity, so as to improve the effect of drug or the other characteristics or property of change drug.Based on pair
The knowledge of drug effect method and drug metabolism in vivo, once pharmaceutical active compounds be it is known, those skilled in the art can set
The pro-drug of compound is counted out (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
It is incorporated by 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 the atomic substitutions with different atomic weight or mass number.It can be with
The example for being introduced into the isotope of these compounds includes, hydrogen, carbon, nitrogen, oxygen, fluorine and chlorine isotope, such as be respectively2H、3H、13C、14C、15N、18O、17O、35S、18F、36Cl.The compound described herein of certain isotope labellings, such as those are therein
Radioactive isotope, such as3H and14C is introduced into, and can be used for measuring drug and/or substrate tissue distribution.Furthermore such as with isotope
Deuterium, i.e.,2The substitution of H can obtain certain treatment advantages, such as increased Half-life in vivo due to the stability of bigger metabolism
Or it reduces and needs 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 and obtains ceiling effect in the analysis that measurement for example inhibits effect as Btk
Amount, concentration or the dosage of fc-specific test FC compound when 50% inhibition.
GI used herein50Refer to and " reaches concentration (the when the 50% of cell proliferation maximum suppression
Concentration for 50%of maximal inhibition of cell proliferation) ".Entirely saying
Among bright book, 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；
Et3N/TEA: triethylamine；
Dioxane:1,4- dioxane；
RT: room temperature；
CH3CN: acetonitrile；
K2CO3: potassium carbonate；
SOCl2: thionyl chloride；
DCM: methylene chloride；
DIEA:N, N- diisopropylethylamine；
HATU:2- (7- aoxidizes benzotriazole)-N, N, N', N'- tetramethylurea hexafluorophosphoric acid ester；
DMF: dimethylformamide；
TFA: trifluoroacetic acid
Step 1:
By m-phenylene diamine (MPD) 1 (0.500g, 4.62mmol) and (Boc)2O (0.92mL, 4.0mmol) is placed in Isosorbide-5-Nitrae-dioxane
In the mixed solution (30mL, volume ratio 2:1) of water, reaction solution is then cooled to 0 degree, be added triethylamine (1.4mL,
10mmol).After end reaction liquid stirs 1 hour under 0 degree, spontaneous recovery to room temperature continues stirring 10 hours at room temperature.It
Afterwards, reaction solution, which is depressurized, is concentrated and is extracted with ethyl acetate, dry with anhydrous magnesium sulfate after organic phase saturated common salt water washing,
Final organic phase is concentrated and uses column chromatography (n-hexane: ethyl acetate=4:1) and obtains compound 2 (0.48g, yield: 58%)
For white solid.
Step 2:
Pyrimidine compound 3 (0.352g, 1.69mmol) and compound 2 (0.270g, 1.69mmol) are mixed in 12mL second
In nitrile, potassium carbonate (0.702g, 5.08mmol) then is added.After reaction solution is stirred at room temperature 3 hours, it is concentrated under reduced pressure, and
Liquid separation in water and ethyl acetate.It is dry with anhydrous magnesium sulfate again after organic phase saturated common salt water washing, it is concentrated under reduced pressure slightly to produce
Product are yellow solid.The crude product and 5%Pd/C are placed in 10mL methanol, after system is emptied air, under an atmosphere of hydrogen
It is stirred at room temperature 5 hours, filters, column chromatography for separation (n-hexane: ethyl acetate=1:1) obtains compound 5 after filtrate concentration
(0.400g, two step yields: 78%), being yellow solid.
Step 3:
3- (trifluoromethyl) benzoic acid 6 (0.500g, 2.63mmol) is placed in 5mL SOCl2In, reaction solution is under stiring
It is heated to reflux to be kept for 1 hour, is then naturally cooling to room temperature, after the dry dilution with toluene of 15mL is added, reaction solution vacuum is dense
Contracting obtains oily residue.After the residue is dissolved in 20mL methylene chloride, 7 (0.478g, 3.16mmol) are added under stiring,
0.2mL triethylamine is then added.After end reaction liquid is stirred at room temperature overnight, be concentrated under reduced pressure, with saturated ammonium chloride solution and
Ethyl acetate liquid separation, it is dry with anhydrous magnesium sulfate again after organic phase saturated common salt water washing.After final organic phase is concentrated under reduced pressure
Column chromatography for separation (n-hexane: ethyl acetate=1:1) obtains compound 8, and (0.70g, yield: 82%) being white solid.
Step 4:
Carboxylic acid 8 (0.263g, 0.813mmol) is placed in 3mL SOCl2In, reaction solution is warming up to reflux under stiring and protects
After holding 1 hour, oily residue is concentrated under reduced pressure to obtain after 10mL dry toluene is added in cooled to room temperature.The grease is molten
5 (0.270g, 0.894mmol) and DIEA (0.10mL, 0.61mmol) are then added in 5mL methylene chloride in solution.Reaction solution is in room
It temperature lower stirring 5 hours, is concentrated under reduced pressure, with saturated sodium bicarbonate solution and ethyl acetate liquid separation, organic phase is washed with saturated common salt
It is dry with anhydrous magnesium sulfate again after washing, it is concentrated under reduced pressure to give crude product, is white solid.The crude product is dissolved in 2mL trifluoro second
In the mixed solution of acid and 2mL methylene chloride, it is concentrated under reduced pressure after being stirred at room temperature 3 hours, with saturated sodium bicarbonate solution and acetic acid
Ethyl ester liquid separation, dry with anhydrous magnesium sulfate again after organic phase saturated common salt water washing, column chromatography for separation (n-hexane: acetic acid second
Ester=1:1) obtaining compound 10, (0.30g, yield: 72%), being yellow solid.
Step 5:
By compound 10 (0.607g, 1.20mmol), 2- cyanoacetic acid (0.204g, 2.40mmol) and HATU (0.912g,
It 3.60mmol) is dissolved in 4mL DMF, TEA (0.50mL, 3.6mmol) then is added.The reaction system was stirred at room temperature
Night.Then, reaction is quenched with saturated sodium bicarbonate aqueous solution, is extracted with ethyl acetate, and is concentrated after being dried with anhydrous magnesium sulfate,
Through the isolated compound 11 of column chromatography (n-hexane: ethyl acetate=1:2), (0.48g, yield: 70%), being white solid.
Step 6:
By compound 11 (0.150g, 0.262mmol), aldehyde R-CHO (0.785mmol) is scattered in 3mL methanol, is then added
Enter piperidines (0.048mL, 0.52mmol), which is placed in tube sealing and is stirred at room temperature overnight.After reaction solution is concentrated under reduced pressure
With ethyl acetate and saturated sodium bicarbonate aqueous solution liquid separation, organic phase saturated common salt water washing, after anhydrous magnesium sulfate drying
It is concentrated under reduced pressure, obtains product RPG-36~RPG-39 through column chromatography for separation (n-hexane: ethyl acetate=2:1), RPG-42~
RPG-47 (yield: 51%~80%).
Synthetic schemes II:
Step 1:
By compound 11 (0.100g, 0.175mmol) and n,N-Dimethylformamide dimethylacetal 12 (0.046mL,
It 0.35mmol) is dissolved in 5mL toluene, after reaction is heated to 90 DEG C of stirrings 3 hours, natural cooling.After solvent under reduced pressure is spin-dried for, second is used
Acetoacetic ester and saturated sodium bicarbonate aqueous solution are extracted, dry with anhydrous magnesium sulfate again after organic phase saturated common salt water washing
It is dry.After final organic phase is concentrated under reduced pressure, product RPG-35 is obtained through column chromatography for separation (n-hexane: ethyl acetate=1:2)
(0.089g, yield: 81%), being white solid.
Synthetic schemes III:
Step 1:
By compound 11 (0.100g, 0.175mmol), aldehyde 13 (0.349mmol) and piperidines (0.032mL, 0.35mmol)
It is scattered in 3mL ethyl alcohol, which is heated to 110 DEG C in tube sealing, reaction solution cooled to room temperature after 2 hours, decompression
With ethyl acetate and saturated sodium bicarbonate aqueous solution liquid separation after concentration, organic phase saturated common salt water washing uses anhydrous magnesium sulfate
After drying, through the isolated 14 (yield: 61%, 70%), solid for white of product of column chromatography (methylene chloride: methanol=20:1)
Body.
Step 2:
Compound 14 (0.150mmol) is placed in the mixed solution of TFA and methylene chloride, at room temperature stirring 3 hours, instead
Liquid reduced pressure is answered to be spin-dried for, with ethyl acetate and 2N NaOH solution liquid separation.After organic phase saturated common salt water washing, then use nothing
Water magnesium sulfate is dry, obtains product RPG-40, RPG-41 through column chromatography for separation (methylene chloride: methanol=10:1) after reduced pressure
(yield: being white solid 83%, 90%).
Synthetic schemes IV:
Step 1:
By compound R PG-49 (0.080g, 0.132mmol), 5- acetylenic acid (0.017ml, 0.16mmol) and HATU
(0.075g, 0.20mmol) is placed in 2mL DMF, and TEA (0.055mL, 0.40mmol) then is added.The reaction is stirred at room temperature
It mixes overnight, after being quenched with saturated sodium bicarbonate aqueous solution, ethyl acetate is added to extract, after organic phase is with saturated common salt water washing, then
It is dry with anhydrous magnesium sulfate, column chromatography for separation (methylene chloride: methanol=20:1) product RPG-50 (0.056g, yield:
It 60%), is light yellow solid.
Test example 1: the external inhibitory activity of Btk and OCI-LY7 cell strain is analyzed
In cell-free kinase assays, the compounds of this invention can be measured with method as described below or the like to Btk
503nhibiting concentration (IC50) and to the 503nhibiting concentration (GI of OCI-LY7 cell strain (its be B cell lymphoma cell strain)50)。
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 R PG-35~RPG-54 is synthesized.
Embodiment compound is characterized as below shown in table 1.In the external inhibitory activity analysis to Btk, implementation of the invention is determined
The IC of example compound R PG-35~RPG-5450Value determines implementation of the invention in the analysis for OCI-LY7 cell strain
The GI of example compound R PG-35~RPG-5450Value.And in table 1 below, give specific IC50And GI50Value.
The characterization and Btk IC of 1 embodiment compound of table50Value and OCI-LY7GI50Value
Test example 2: the covalent reversible 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 0.5% dimethyl sulfoxide (as control) mixing of concentration shown in 1.0ngBtk kinases and following table, room
After temperature is incubated for 30 minutes, a part of mixture is taken out, after 1 × reaction buffer 2 × dilution, 1 μM of kit is added and is provided
Substrate and 20 μM of atriphos add after reaction 1 hour according to the step of record in HTRF tyrosine kinase kit specification
Enter detection reagent termination, microplate reader (Perkin Elmer Envision) reads data.The examination of 0.5% dimethyl sulfoxide is added
Sample is as control, with its kinase activity under same incubation time for 100%, to through each compound treated kinase activity
It is normalized, the kinase inhibition rate of each compound is calculated, as shown in table 1 and Fig. 1.
1 bruton's tyrosine kinase maximum inhibition percentage % of table
Experiments have shown that compound R PG41 to the active inhibiting rate of bruton's tyrosine kinase with the dilution of reaction system and
It reduces, this is because dilution makes it dissociate to reduce the inhibiting effect to kinases with kinases, embodies covalent reversible combination
Activity；And control compounds are substantially unchanged for Buddhist nun's inhibiting rate according to Shandong.
Test example 3: the covalent reversible inhibitory activity of compound (fluorescence probe combines test)
Reaction buffer is 1 × PBS (Phosphate Buffer Saline, GIBCO, article No. C10010500CP), instead
Answering volume is 20 μ L.Mixing respectively according to Shandong for Buddhist nun or RPG41 for 1480ng Btk kinases and 1% dimethyl sulfoxide or 1 μM of concentration is taken,
After being incubated for 15 minutes, take out appropriate volume, dilute 1 respectively with 1 × PBS ×, 3 ×, 5 ×, 10 ×, volume is 9uL after dilution, is added
Enter reported covalent fluorescence probe [Scientific Reports, 2015,5,16136] 1 μ L, makes final concentration of 0.5 μ of probe
M, reaction volume are 10 μ L, and the 2X loading buffer that equivalent responses volume is added after reaction 5 minutes, 15 minutes is terminated instead
It answers.20 μ L of end reaction volume.Then with 1X loading buffer by 1X, 3X, 5X, 10X sample dilutes 10X respectively,
3.33X, 2X, 1X, making the enzyme concentration of each processing is uniformly 3.33ng/ μ L, respectively takes 10 μ L loadings, carries out after SDS-PAGE electrophoresis
Fluorescent scanning carries out Western blot detection with Btk monoclonal antibody (CST, article No. 8547), verifies the sample of electrophoresis loading
Whether middle enzyme concentration is uniform.As a result as shown in Fig. 2, the Western blot band of BTK is uniform, illustrate that the Btk kinases of loading is dense
Degree is consistent；The Btk band of 1%DMSO processing shows strong fluorescence, illustrates fluorescence probe pass flag Btk；And compound R PG41
Swimming lane in, with the increase of extension rate, the fluorescence intensity of band is gradually increased, and illustrates compound R PG41 with the dilute of mixed liquor
It releases and is dissociated from kinases, extension rate is bigger, and RPG41 is dissociated more, and causing can be with the Btk kinases in conjunction with fluorescence probe
It is more, therefore corresponding band fluorescence is stronger, the covalent reversible for embodying compound combines activity；And control compounds are according to Shandong
For this unstressed configuration of Thessaloniki, illustrate not dissociate after replacing Buddhist nun and kinases covalent bond according to Shandong.
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
It is incorporated by herein with for all purposes.
Claims (11)
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 or heteroaryl: halogen, ammonia
Base, C1-6Alkyl, C1-6Alkoxy, halogenated C1-6Alkyl, C1-6Alkyl-heterocyclyl groups-C1-3Alkyl,
N is 0 or 1,
X is selected from H, halogen or C1-6Alkyl,
R is amino, the C optionally replaced by 1,2 or 3 substituent group selected from the following1-6Alkyl, C3-8Naphthenic base, aryl or miscellaneous
Aryl: halogen, hydroxyl, amino, C1-6Alkyl, C1-6Alkyl amino, C1-6Alkoxy, C2-6Alkynyl-C1-3Alkyl-(CO-NH)-.
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, methoxyl group, ethyoxyl, propoxyl group, CF by 1,2 or 33OrSubstituent group replace cyclopropyl, phenyl, naphthalene,
N is 1.
3. compound as claimed in claim 1 or 2 or its pharmaceutically acceptable salt, wherein X is selected from H, F, Cl or methyl.
4. compound according to any one of claims 1 to 3 or its pharmaceutically acceptable salt, wherein R is optionally
F, Cl, Br, hydroxyl, amino, methyl, ethyl, methylamino, ethylamino, methoxyl group, ethyoxyl, acetenyl-are selected from by 1,2 or 3
The amino of the substituent group substitution of propyl-(CO-NH)-, cyclopropyl, isopropyl, tert-butyl, phenyl, furyl, imidazole radicals, thiophene
Base, pyrrole radicals, pyridyl group.
5. compound as claimed in claim 4 or its pharmaceutically acceptable salt, wherein R is optionally to be selected by 1,2 or 3
Amino, cyclopropyl, isopropyl from the substituent group substitution of F, hydroxyl, amino, methyl, methylamino, acetenyl-propyl-(CO-NH)-
Base, tert-butyl, phenyl, furyl, imidazole radicals, thienyl.
6. a kind of compound, is selected from:
7. such as compound according to any one of claims 1 to 6, for inhibiting bruton's tyrosine kinase active.
8. compound as claimed in claim 7, the compound can inhibit bruton's tyrosine to swash in a manner of covalent bond
Enzymatic activity.
9. such as compound according to any one of claims 1 to 8, the compound can on bruton's tyrosine kinase
Amino acid residue forms reversible covalent bonds.
10. a kind of Pharmaceutical composition, it includes the compounds and medicine according to any one of claims 1 to 9 of therapeutically effective amount
Acceptable carrier or excipient on.
11. compound according to any one of claims 1 to 9 or its pharmaceutically acceptable salt are following for treating in preparation
Purposes in the drug of disease or situation: autoimmune disease, heteroimmunity disease, inflammation disease, cancer, thromboembolism
Disease.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108191871A (en) * | 2018-01-02 | 2018-06-22 | 成都倍特药业有限公司 | Novel cloth Shandong tyrosine kinase inhibitor and its preparation method and application |
| WO2020239103A1 (en) * | 2019-05-31 | 2020-12-03 | 四川海思科制药有限公司 | Btk inhibitor ring derivative, preparation method therefor and pharmaceutical application thereof |
| CN113234026A (en) * | 2021-03-28 | 2021-08-10 | 北京大学深圳研究生院 | Compound with B lymphocyte tyrosine kinase inhibitory activity and application thereof |
| CN114085207A (en) * | 2020-10-16 | 2022-02-25 | 广州百霆医药科技有限公司 | Bruton tyrosine protein kinase inhibitor and application thereof |
| CN113234026B (en) * | 2021-03-28 | 2024-04-30 | 北京大学深圳研究生院 | Compound with B lymphocyte tyrosine kinase inhibitory activity and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103073508A (en) * | 2011-10-25 | 2013-05-01 | 北京大学深圳研究生院 | Kinase inhibitor and method for treating related diseases |
| CN103694268A (en) * | 2013-12-16 | 2014-04-02 | 李锡涛 | Kinase activity probe and method for treating related diseases |
| CN104109127A (en) * | 2013-04-19 | 2014-10-22 | 北京大学深圳研究生院 | Kinases inhibitor and method for treating correlated diseases |
-
2017
- 2017-09-04 CN CN201710787857.6A patent/CN109422696B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103073508A (en) * | 2011-10-25 | 2013-05-01 | 北京大学深圳研究生院 | Kinase inhibitor and method for treating related diseases |
| CN104109127A (en) * | 2013-04-19 | 2014-10-22 | 北京大学深圳研究生院 | Kinases inhibitor and method for treating correlated diseases |
| CN103694268A (en) * | 2013-12-16 | 2014-04-02 | 李锡涛 | Kinase activity probe and method for treating related diseases |
Non-Patent Citations (1)
| Title |
|---|
| XITAO LI等: "Discovery of a Series of 2,5-Diaminopyrimidine Covalent Irreversible Inhibitors of Bruton’s Tyrosine Kinase with in Vivo Antitumor Activity", 《J. MED. CHEM.》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108191871A (en) * | 2018-01-02 | 2018-06-22 | 成都倍特药业有限公司 | Novel cloth Shandong tyrosine kinase inhibitor and its preparation method and application |
| WO2020239103A1 (en) * | 2019-05-31 | 2020-12-03 | 四川海思科制药有限公司 | Btk inhibitor ring derivative, preparation method therefor and pharmaceutical application thereof |
| TWI762939B (en) * | 2019-05-31 | 2022-05-01 | 大陸商海思科醫藥集團股份有限公司 | BTK inhibitor cyclic derivatives, preparation method and pharmaceutical application thereof |
| US11542266B1 (en) | 2019-05-31 | 2023-01-03 | Haisco Pharmaceuticals Pte. Ltd. | Substituted piperidines as BTK inhibitors |
| CN114085207A (en) * | 2020-10-16 | 2022-02-25 | 广州百霆医药科技有限公司 | Bruton tyrosine protein kinase inhibitor and application thereof |
| CN113234026A (en) * | 2021-03-28 | 2021-08-10 | 北京大学深圳研究生院 | Compound with B lymphocyte tyrosine kinase inhibitory activity and application thereof |
| CN113234026B (en) * | 2021-03-28 | 2024-04-30 | 北京大学深圳研究生院 | Compound with B lymphocyte tyrosine kinase inhibitory activity and application thereof |
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| CN109422696B (en) | 2020-10-30 |
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