WO2020087565A1 - 吲唑类激酶抑制剂及其用途 - Google Patents

吲唑类激酶抑制剂及其用途 Download PDF

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WO2020087565A1
WO2020087565A1 PCT/CN2018/115025 CN2018115025W WO2020087565A1 WO 2020087565 A1 WO2020087565 A1 WO 2020087565A1 CN 2018115025 W CN2018115025 W CN 2018115025W WO 2020087565 A1 WO2020087565 A1 WO 2020087565A1
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optionally substituted
ckit
kinase inhibitor
methyl
groups
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PCT/CN2018/115025
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English (en)
French (fr)
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刘静
刘青松
刘学松
王蓓蕾
陈程
齐紫平
王文超
王俊杰
蒋宗儒
汪文亮
亓爽
王黎
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安徽中科拓苒药物科学研究有限公司
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Priority to EP18938587.5A priority Critical patent/EP3878841B1/en
Priority to AU2018447240A priority patent/AU2018447240B2/en
Priority to US17/288,966 priority patent/US20220002287A1/en
Priority to JP2021523586A priority patent/JP7176798B2/ja
Publication of WO2020087565A1 publication Critical patent/WO2020087565A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond

Definitions

  • the present application relates to a kinase inhibitor, as well as methods and uses of such kinase inhibitors to inhibit kinase activity. More specifically, the present invention relates to inhibitors capable of inhibiting the activity of cKIT (particularly mutant cKIT / T670I), FLT3 (including mutant FLT3-ITD), PDGFR ⁇ , PDGFR ⁇ , and / or VEGFR2 kinase.
  • Gastrointestinal stromal tumors are the most common mesenchymal tumors of the gastrointestinal tract. The incidence of GIST is about 1 / 100,000 to 2 / 100,000, accounting for 1-3% of all gastrointestinal tumors. The disease is more common in middle-aged and elderly people. The median age of onset is 50 to 65 years old. It is rare before 40 years old, but it has also been reported in children. GIST is currently considered to be a tumor with potentially malignant behavior, and its biological behavior is difficult to predict.
  • GIST can occur in any part of the digestive tract, the most common is the stomach (60% to 70%), followed by the small intestine (20% to 30%), and the incidence of esophagus, colon, and rectum is less than 10%, and it can also occur in the net Membranes and mesentery.
  • cKIT mutant 80-85%
  • PDGFR ⁇ mutant 5-10%
  • cKIT wild-type GISTs 10 %
  • the pathogenesis of gastrointestinal stromal tumors is related to the activation of cKIT protein (CD117) signaling pathway.
  • the proto-oncogene cKIT is a homolog of the vKIT gene isolated from feline fibrosarcoma virus. It is located on human chromosome 4 (4q12-13) and is about 90 kb in length. It consists of 21 exons and 20 endogenous The sub-composition is highly conservative during evolution.
  • the cKIT protein is a receptor tyrosine kinase (RTK) located on the cell membrane. Its relative molecular mass is 145000, and it is named CD117 according to its cell surface epitope.
  • the cKIT protein belongs to the third type RTK family, consisting of 5 immunoglobulin-like domains (D1 ⁇ D5), 1 transmembrane domain, and 1 containing the proximal membrane domain (JMD) and tyrosine kinase (TK) The cytoplasmic region of the domain constitutes.
  • the TK domain is further divided into adenosine triphosphate (ATP) domain (TK1) and phosphotransferase (phosphotransferase) domain (TK2).
  • the ligand stem cell factor SCF (stem cell) combines with the extracellular domain to form a dimer, resulting in the autophosphorylation of tyrosine in the TK domain of the cytoplasmic region, which further causes the autophosphorylation of various downstream effects and completes various signals Of passing.
  • the main signaling pathways include PI3K signaling pathway, JAK-STAT signaling pathway, Ras-Erk signaling pathway, Src family kinase signaling pathway and PLC signaling pathway, etc., which ultimately promotes cell proliferation, division and tissue growth and survival.
  • the cKIT kinase inhibitors used in the clinical treatment of GIST so far mainly include Imatinib (cKIT / BCR-ABL / PDGFR) of Novartis and Sunitinib (cKIT / BCR-ABL / PDGFR) of Pfizer / VEGFR2 / FLT3).
  • Imatinib is the first type II kinase inhibitor for the treatment of GIST
  • Sunitinib is a type I cKIT kinase inhibitor approved by the FDA in 2006.
  • type II kinase inhibitors with cKIT activity include Regorafenib, Nilotinib, Mastinib, Sorafenib, etc. These small molecule inhibitors are all multi-target inhibitors, and Nilotinib and Mastinib cannot overcome the T670I mutation. Although two small molecule inhibitors Regorafenib and Sorafenib with cKIT activity developed by Bayer have certain activity on the cKIT-T670I mutation, both are multi-target inhibitors.
  • Axitinib is a multi-target small molecule inhibitor developed by Pfizer, and also has inhibitory activities of cKIT, BCR-ABL, VEGFR2 and other kinases.
  • Ponatinib also has a strong inhibitory activity against cKIT kinase, and it can overcome the resistance of cKIT-T670I mutation, but it is ABL / PDGFR / RET / CSF1R / FGFR / VEGFR / FGFR / RET And other multi-target inhibitors of kinases.
  • the present invention is mainly to find an inhibitor with a completely new structure and a strong inhibitory effect on the imatinib resistance mutation cKIT-T670I.
  • the present invention provides a selective kinase inhibitor, including a compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug thereof:
  • Y is selected from -NH- or-(CH 2 ) n- , where n is an integer of 0-3;
  • R 1 is selected is optionally substituted with 1-3 independent R 4 groups are phenyl, optionally substituted with 1-3 independent R 4 groups pyridyl, optionally substituted with 1-3 Pyrazolyl substituted with independent R 4 groups, and pyrimidinyl optionally substituted with 1-3 independent R 4 groups;
  • R 2 is selected from hydrogen and C 1-6 alkyl
  • R 3 is selected from C 1-6 alkyl, C 1-6 alkylamino, optionally substituted with 1-2 independent R 5 groups, and optionally 1-3 independent R 4 groups substituted phenyl, optionally substituted with 1-3 independent R 4 groups naphthyl, optionally substituted with 1-3 independent R 4 groups pyridyl, optionally substituted with 1-3 Piperazinyl substituted with an independent R 4 group, and piperidinyl optionally substituted with 1-3 independent R 4 groups;
  • R 4 is independently selected from halogen, amino, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 1 -6 alkylamino, C 2-6 alkylamido, (4-methylpiperazin-1-yl) methyl, morpholinomethyl, morpholinyl, 4-methylpiperazin-1-yl, 4 -Piperidinyl, and 4-tetrahydropyranyl;
  • R 5 is independently selected from amino, hydroxyl, and C 1-6 alkylthio.
  • Y is a direct bond or -CH 2- .
  • R 1 is selected from phenyl, pyridyl, pyrazolyl, and pyrimidinyl optionally substituted with 1-3 independent R 4 groups, wherein R 4 is independently selected from halogen , Amino, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, and (4-methylpiperazin-1-yl) methyl; R 1 is more preferably optionally Methyl, amino or halogen substituted phenyl, 2-pyridyl, 3-pyridyl, 4-pyrazolyl, and 5-pyrimidinyl; R 1 is particularly preferably 2-pyridyl.
  • R 2 is hydrogen or methyl
  • R 3 is selected from C 1-6 alkyl, C 1-6 alkylamino, optionally substituted by 1-2 independent R 5 groups, and optionally 1- 3 R 4 groups independently substituted with phenyl, naphthyl, pyridyl, piperazinyl, and piperidinyl, wherein R 4 is independently selected from halogen, amino, C 1-6 alkyl, C 1-6 Haloalkyl, C 1-6 alkoxy, and (4-methylpiperazin-1-yl) methyl, R 5 is independently selected from amino, hydroxyl, and methylthio; R 3 is more preferably optionally C 1-6 alkyl substituted by amino, hydroxy, or methylthio, dimethylamino, N-piperazinyl optionally substituted by methyl, optionally halogen, trifluoromethyl, or methoxy Group substituted phenyl, naphthyl, 4-pyridyl, 3-piperidinyl, and 4-piperidinyl optionally substituted
  • R 3 when Y is a direct bond, R 3 is selected from C 1-6 alkyl, and 4-pyridyl optionally substituted with amino, hydroxy, or methylthio; when Y is- When CH 2- , R 3 is selected from phenyl optionally substituted with methoxy, N-piperazinyl optionally substituted with methyl, and 4-piperidinyl optionally substituted with methyl.
  • the invention also relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the kinase inhibitor of the invention, and a pharmaceutically acceptable carrier or excipient, and optionally other therapeutic agents.
  • the invention also relates to the use of kinase inhibitors or pharmaceutical compositions including them to reduce or inhibit cKIT (especially mutant cKIT / T670I), FLT3 (including mutant FLT3-ITD), PDGFR ⁇ , PDGFR ⁇ , and / or VEGFR2 kinase activity methods and uses.
  • the present invention also relates to the prevention or treatment of cKIT (especially mutant cKIT / T670I), FLT3 (including mutant FLT3-ITD), PDGFR ⁇ in a subject using kinase inhibitors or pharmaceutical compositions including the same , PDGFR ⁇ , and / or VEGFR2 activity-related disorders methods and uses.
  • cKIT especially mutant cKIT / T670I
  • FLT3 including mutant FLT3-ITD
  • PDGFR ⁇ in a subject using kinase inhibitors or pharmaceutical compositions including the same , PDGFR ⁇ , and / or VEGFR2 activity-related disorders methods and uses.
  • Figure 1a shows the effect of compound 9 and sunitinib on the body weight of mice after administration in a tel-cKIT / T670I-BaF3 cell tumor transplantation mouse model
  • Figure 1b shows compound 9 and sunitinib in tel- Tumor inhibition effect in cKIT / T670I-BaF3 cell tumor transplantation mouse model.
  • Figure 2a shows the effect of compound 9 and sunitinib on the body weight of mice after administration in the GIST-T1-T670I cell tumor transplantation mouse model
  • Figure 2b shows compound 9 and sunitinib in GIST-T1- Tumor suppressive effect in T670I cell tumor transplantation mouse model.
  • the present invention uses conventional methods such as mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA technology, and pharmacology within the technical scope of the art.
  • mass spectrometry NMR, HPLC, protein chemistry, biochemistry, recombinant DNA technology, and pharmacology
  • nomenclature and laboratory operations and techniques that are chemically related to analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry described herein are known to those skilled in the art.
  • the aforementioned techniques and steps can be implemented by conventional methods well known in the art and described in various general and more specific documents, which are cited and discussed in this specification.
  • alkyl refers to an aliphatic hydrocarbon group, which may be a branched or linear alkyl group. Depending on the structure, the alkyl group may be a monovalent group or a divalent group (ie, an alkylene group). In the present invention, the alkyl group is preferably an alkyl group having 1 to 8 carbon atoms, more preferably a "lower alkyl group” having 1 to 6 carbon atoms, and even more preferably an alkyl group having 1 to 4 carbon atoms. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, and the like.
  • alkyl mentioned herein includes all possible configurations and conformations of the alkyl group, for example, the "propyl” mentioned herein includes n-propyl and isopropyl, and the "butyl” includes n-butyl Group, isobutyl and tert-butyl, "pentyl” includes n-pentyl, isopropyl, neopentyl, tert-pentyl, and pent-3-yl and so on.
  • alkoxy refers to -O-alkyl, where alkyl is as defined herein. Typical alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy and the like.
  • alkoxyalkyl means that the alkyl group defined herein is substituted with an alkoxy group defined herein.
  • cycloalkyl refers to a monocyclic or polycyclic group that contains only carbon and hydrogen. Cycloalkyl includes groups having 3-12 ring atoms. According to the structure, the cycloalkyl group may be a monovalent group or a divalent group (for example, cycloalkylene group). In the present invention, the cycloalkyl group is preferably a cycloalkyl group having 3 to 8 carbon atoms, and more preferably a "lower cycloalkyl group" having 3 to 6 carbon atoms.
  • cycloalkyl examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and adamantane base.
  • alkyl (cycloalkyl) or "cycloalkylalkyl” means that the alkyl group defined herein is substituted with a cycloalkyl group defined herein.
  • Non-limiting cycloalkylalkyl groups include cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, and the like.
  • aromatic group refers to a planar ring having a delocalized ⁇ electron system and contains 4n + 2 ⁇ electrons, where n is an integer.
  • the aromatic ring may be composed of five, six, seven, eight, nine, or more than nine atoms.
  • the aromatic group may be optionally substituted.
  • aryl includes carbocyclic aryl (eg phenyl) and heterocyclic aryl (or “heteroaryl” or “heteroaryl”) groups (eg pyridine).
  • the term includes monocyclic or fused-ring polycyclic (ie, rings that share adjacent pairs of carbon atoms) groups.
  • aryl means that each atom in the aryl ring that constitutes the ring is a carbon atom.
  • the aryl ring may be composed of five, six, seven, eight, nine, or more than nine atoms.
  • the aryl group may be optionally substituted. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, phenanthrenyl, anthracenyl, fluorenyl, and indenyl.
  • the aryl group may be a monovalent group or a divalent group (ie, an arylene group).
  • aryloxy refers to -O-aryl, where aryl is as defined herein.
  • heteroaryl means that the aryl group includes one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the N-containing “heteroaryl” moiety means that at least one skeleton atom in the ring of the aromatic group is a nitrogen atom.
  • the heteroaryl group may be a monovalent group or a divalent group (ie, a heteroarylene group).
  • heteroaryl groups include, but are not limited to pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazole Group, isothiazolyl group, pyrrolyl group, quinolinyl group, isoquinolinyl group, indolyl group, benzimidazolyl group, benzofuranyl group, indazolyl group, indazine group, phthalazinyl group, pyridazinyl group, isoindolin Indolyl, pteridyl, purinyl, oxadiazolyl, thiadiazolyl, furazyl, benzofurazyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl , Nap
  • alkyl (aryl) or “aralkyl” means that the alkyl group defined herein is substituted with an aryl group defined herein.
  • Non-limiting alkyl (aryl) groups include benzyl, phenethyl and the like.
  • alkyl (heteroaryl) or “heteroarylalkyl” means that the alkyl group defined herein is substituted with a heteroaryl group defined herein.
  • heteroalkyl means that one or more of the backbone chain atoms in the alkyl group defined herein are heteroatoms, such as oxygen, nitrogen, sulfur, silicon, phosphorus, or a combination thereof.
  • the heteroatom (s) can be located at any position inside the heteroalkyl group or at a position where the heteroalkyl group is connected to the rest of the molecule.
  • heterocycloalkyl refers to a non-aromatic ring in which one or more atoms constituting the ring are heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the heterocycloalkyl ring may be composed of three, four, five, six, seven, eight, nine, or more than nine atoms.
  • the heterocycloalkyl ring may be optionally substituted.
  • heterocycloalkyl groups include, but are not limited to, lactams, lactones, cyclic imines, cyclic thioimines, cyclic carbamates, tetrahydrothiopyran, 4H-pyran, tetrahydropyran, piperidine, 1,3-dioxin, 1,3-dioxane, 1,4-dioxane, 1,4-dioxane, piperazine, 1,3-oxathiolane, 1,4- Oxythiacyclohexadiene, 1,4-oxathiolane, tetrahydro-1,4-thiazine, 2H-1,2-oxazine, maleimide, succinimide, barium Bituric acid, thiobarbituric acid, dioxopiperazine, hydantoin, dihydrouracil, morpholine, trioxane, hexahydro-1,3,5-triazine, t
  • alkyl (heterocycloalkyl) or “heterocycloalkylalkyl” means that the alkyl group defined herein is substituted with a heterocycloalkyl group defined herein.
  • alkoxy (heterocycloalkyl) or “heterocycloalkylalkoxy” means that the alkoxy group defined herein is substituted with a heterocycloalkyl group defined herein.
  • halo or halogen refers to fluorine, chlorine, bromine and iodine.
  • haloalkyl examples include structures of alkyl, alkoxy or heteroalkyl groups in which at least one hydrogen is replaced by a halogen atom. In some embodiments, if two or more hydrogen atoms are replaced with halogen atoms, the halogen atoms are the same as or different from each other.
  • hydroxyl refers to the -OH group.
  • cyano refers to a -CN group.
  • ester group refers to a chemical moiety having the formula -COOR, where R is selected from alkyl, cycloalkyl, aryl, heteroaryl (attached through a ring carbon) and heterocyclic group (attached through a ring carbon).
  • amino refers to the -NH 2 group.
  • aminoacyl refers to a -CO-NH 2 group.
  • amido or “amido” refers to -NR-CO-R ', where R and R' are each independently hydrogen or alkyl.
  • alkylamino refers to an amino substituent that is further substituted with one or two alkyl groups, specifically refers to the group -NRR ', where R and R' are each independently selected from hydrogen or lower alkyl, provided that- NRR 'is not -NH 2 .
  • Alkyl amino includes groups wherein the nitrogen -NH 2 group is connected to at least one compound of an alkyl group. Examples of alkylamino groups include, but are not limited to, methylamino, ethylamino, and the like.
  • “Dialkylamino” includes groups in which the nitrogen of -NH 2 connects at least two other alkyl groups. Examples of dialkylamino groups include, but are not limited to, dimethylamino, diethylamino, and the like.
  • arylamino and diarylamino refer to amino substituents that are further substituted with one or two aryl groups, specifically referring to the group -NRR ', where R and R' are each independently selected from hydrogen, Lower alkyl, or aryl, where N connects at least one or two aryl groups, respectively.
  • cycloalkylamino refers to an amino substituent that is further substituted with one or two cycloalkyl groups as defined herein.
  • heteroalkylamino refers to an amino substituent that is further substituted with one or two heteroalkyl groups as defined herein.
  • aralkylamino herein refers to a group -NRR 'where R is lower aralkyl and R' is hydrogen, lower alkyl, aryl or lower aralkyl.
  • heteroarylamino refers to an amino substituent that is further substituted with one or two heteroaryl groups as defined herein.
  • heterocycloalkylamino means that the amino group defined herein is substituted with a heterocycloalkyl group defined herein.
  • alkylaminoalkyl means that the alkyl group defined herein is substituted with an alkylamino group defined herein.
  • aminoalkyl refers to an alkyl substituent that is further substituted with one or more amino groups.
  • aminoalkoxy refers to an alkoxy substituent that is further substituted with one or more amino groups.
  • hydroxyalkyl or "hydroxyalkyl” refers to an alkyl substituent that is further substituted with one or more hydroxyl groups.
  • cyanoalkyl refers to an alkyl substituent that is further substituted with one or more cyano groups.
  • acyl refers to a monovalent radical remaining after removing the hydroxyl group of an organic or inorganic oxyacid, the general formula is R-M (O)-, where M is usually C.
  • alkanoyl or “alkylcarbonyl” refers to a carbonyl group that is further substituted with an alkyl group.
  • Typical alkanoyl groups include, but are not limited to, acetyl, propionyl, butyryl, valeryl, hexanoyl, and the like.
  • arylcarbonyl means that the carbonyl group defined herein is substituted with an aryl group defined herein.
  • alkoxycarbonyl refers to a carbonyl group that is further substituted with an alkoxy group.
  • heterocycloalkylcarbonyl refers to a carbonyl group that is further substituted with a heterocycloalkyl group.
  • alkylaminocarbonyl cycloalkylaminocarbonyl
  • arylaminocarbonyl aralkylaminocarbonyl
  • heteroarylaminocarbonyl respectively mean that the carbonyl groups defined herein are respectively defined herein Alkylamino, cycloalkylamino, arylamino, aralkylamino, or heteroarylamino substitution.
  • alkylcarbonylalkyl or “alkanoylalkyl” refers to an alkyl group further substituted with an alkylcarbonyl group.
  • alkylcarbonylalkoxy or “alkanoylalkoxy” refers to an alkoxy group further substituted with an alkylcarbonyl group.
  • heterocycloalkylcarbonylalkyl refers to an alkyl group further substituted with a heterocycloalkylcarbonyl group.
  • mercapto refers to a -SH group.
  • alkylthio means that the mercapto group defined herein is substituted with an alkyl group defined herein.
  • alkylaminosulfone group means that the sulfone group defined herein is substituted with an alkylamino group defined herein.
  • alkylsulfonylamino or "cycloalkylsulfonylamino” means that the amino group defined herein is substituted with an alkylsulfonyl group or cycloalkylsulfone group as defined herein.
  • quaternary ammonium group refers to -N + RR'R ", wherein R, R 'and R" are each independently selected from alkyl groups having 1-8 carbon atoms.
  • optional means that one or more of the events described below may or may not occur, and includes both events that occur and events that do not occur.
  • optionally substituted or “substituted” means that the mentioned group may be substituted with one or more additional groups, each of which is independently and independently selected from alkyl, cycloalkyl , Aryl, heteroaryl, heterocyclic, hydroxy, alkoxy, cyano, halogen, amido, nitro, haloalkyl, amino, methanesulfonyl, alkylcarbonyl, alkoxycarbonyl, heteroaryl Alkyl, heterocycloalkylalkyl, aminoacyl, amino protecting group, etc.
  • the amino protecting group is preferably selected from pivaloyl, tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyl, p-methoxybenzyl, allyloxycarbonyl, trifluoroacetyl and the like.
  • tyrosine protein kinase is a type of kinase that catalyzes the transfer of ⁇ -phosphate from ATP to protein tyrosine residues and can catalyze a variety of substrate protein tyrosine residues Phosphorylation plays an important role in cell growth, proliferation and differentiation.
  • the term “inhibition”, “inhibited”, or “inhibitor” of a kinase refers to the inhibition of phosphotransferase activity.
  • the "metabolite” of a compound disclosed herein is a derivative of the compound that is formed when the compound is metabolized.
  • active metabolite refers to a biologically active derivative of a compound that is formed when the compound is metabolized.
  • the term “metabolized” refers to the sum of processes by which specific substances are changed by an organism (including but not limited to hydrolysis reactions and reactions catalyzed by enzymes, such as oxidation reactions). Therefore, enzymes can produce specific structural transformations into compounds.
  • cytochrome P450 catalyzes various oxidation and reduction reactions
  • glucosyl diphosphate transferase catalyzes the conversion of activated glucuronic acid molecules to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines, and free sulfhydryl groups.
  • Metabolites of the compounds disclosed herein can be identified by administering the compound to a host and analyzing tissue samples from the host, or by incubating the compound with hepatocytes in vitro and analyzing the resulting compound. Both methods are known in the art.
  • the metabolite of the compound is formed by an oxidation process and corresponds to the corresponding hydroxyl-containing compound.
  • the compound is metabolized to a pharmaceutically active metabolite.
  • the term "modulate" refers to directly or indirectly interacting with a target to change the activity of the target, including, by way of example only, enhancing the activity of the target, inhibiting the activity of the target, limiting the activity of the target, or extending the activity of the target.
  • target protein refers to a protein molecule or part of a protein that can be bound by a selective binding compound.
  • the target protein is tyrosine kinase KIT (wild type or various mutations or combinations thereof), ABL (wild type or various mutations or combinations thereof), EGFR (wild type or various mutations or combinations thereof) Combination), FLT3 (wild type or various mutations or combinations thereof), VEGFR2 (wild type or various mutations or combinations thereof), RET (wild type or various mutations or combinations thereof), PDGFR ⁇ (wild type or various mutations) (Or combinations thereof), PDGFR ⁇ (wild type or various mutations or combinations thereof), BCR / ABL (wild type or various mutations or combinations thereof), FGFR1 (wild type or various mutations or combinations thereof), FGFR2 (wild type Or various mutations or combinations thereof), FGFR3 (wild type or various mutations or combinations thereof), FGFR4 (wild type or various mutations or combinations thereof).
  • IC 50 as used herein refers to the amount, concentration, or dose of a particular test compound that achieves 50% inhibition of the maximum effect in an analysis measuring such effects.
  • EC 50 refers to the dose, concentration, or amount of an assay compound that causes a dose-dependent response of 50% of the maximum expression of a specific response induced, stimulated, or potentiated by a particular assay compound.
  • GI 50 refers to the concentration of the drug required to inhibit the growth of 50% of the cells, that is, the concentration of the drug when the drug inhibits or controls the growth of 50% of the cells (such as cancer cells).
  • novel kinase inhibitor of the present invention is derived from the novel kinase inhibitor of the present invention.
  • the present invention provides a novel kinase inhibitor, including a compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug thereof,
  • Y is selected from -NH- or-(CH 2 ) n- , where n is an integer of 0-3, and when n is 0, Y represents a direct bond;
  • R 1 is selected from aryl and heteroaryl optionally substituted with 1-3 independent R 4 groups;
  • R 2 is selected from hydrogen and C 1-6 alkyl
  • R 3 is selected from C 1-6 alkyl, C 1-6 alkylamino, optionally substituted with 1-2 independent R 5 groups, and optionally 1-3 independent R 4 groups Substituted aryl, heteroaryl and heterocyclic groups;
  • R 4 is independently selected from halogen, amino, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 1 -6 alkylamino, C 2-6 alkylamido, (4-methylpiperazin-1-yl) methyl, morpholinomethyl, morpholinyl, 4-methylpiperazin-1-yl, 4 -Piperidinyl, and 4-tetrahydropyranyl;
  • R 5 is independently selected from amino, hydroxyl, and C 1-6 alkylthio.
  • Y is a direct bond or -CH 2- .
  • R 1 is selected from phenyl, pyridyl, pyrazolyl, and pyrimidinyl optionally substituted with 1-3 independent R 4 groups, wherein R 4 is independently selected from halogen , Amino, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, and (4-methylpiperazin-1-yl) methyl; R 1 is more preferably optionally Methyl, amino or halogen substituted phenyl, 2-pyridyl, 3-pyridyl, 4-pyrazolyl, and 5-pyrimidinyl; R 1 is particularly preferably 2-pyridyl.
  • R 2 is hydrogen or methyl
  • R 3 is selected from C 1-6 alkyl, C 1-6 alkylamino, optionally substituted by 1-2 independent R 5 groups, and optionally 1- 3 R 4 groups independently substituted with phenyl, naphthyl, pyridyl, piperazinyl, and piperidinyl, wherein R 4 is independently selected from halogen, amino, C 1-6 alkyl, C 1-6 Haloalkyl, C 1-6 alkoxy, and (4-methylpiperazin-1-yl) methyl, R 5 is independently selected from amino, hydroxyl, and methylthio; R 3 is more preferably optionally C 1-6 alkyl substituted by amino, hydroxy, or methylthio, dimethylamino, N-piperazinyl optionally substituted by methyl, optionally halogen, trifluoromethyl, or methoxy Group substituted phenyl, naphthyl, 4-pyridyl, 3-piperidinyl, and 4-piperidinyl optionally substituted
  • R 3 when Y is a direct bond, R 3 is selected from C 1-6 alkyl, and 4-pyridyl optionally substituted with amino, hydroxy, or methylthio; when Y is- When CH 2- , R 3 is selected from phenyl optionally substituted with methoxy, N-piperazinyl optionally substituted with methyl, and 4-piperidinyl optionally substituted with methyl.
  • the inhibitors of the present invention include the compounds of Table 1 below or pharmaceutically acceptable salts, solvates, esters, acids, metabolites, or prodrugs thereof.
  • novel kinase inhibitors are also described herein. Also described herein are pharmaceutically acceptable salts, solvates, esters, acids, pharmaceutically active metabolites, and prodrugs of this compound.
  • the compound described herein is metabolized in the body of an organism in need thereof to produce a metabolite, and the generated metabolite is then used to produce a desired effect, including a desired therapeutic effect.
  • the compounds described herein can be made and / or used as pharmaceutically acceptable salts.
  • the types of pharmaceutically acceptable salts include, but are not limited to: (1) acid addition salts, formed by reacting the free base form of the compound with pharmaceutically acceptable inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, Nitric acid, phosphoric acid, metaphosphoric acid, etc .; or formed by reaction with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, lemon Acid, succinic acid, maleic acid, tartaric acid, fumaric acid, trifluoroacetic acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid Acid, 1,2-ethanedisulfonic acid, 2-
  • the corresponding counter ion of the pharmaceutically acceptable salt can be analyzed and identified using various methods including, but not limited to, ion exchange chromatography, ion chromatography, capillary electrophoresis, inductively coupled plasma, atomic absorption spectroscopy, mass spectrometry, or any of them combination.
  • the salt is recovered using at least one of the following techniques: filtration, precipitation with a non-solvent followed by filtration, solvent evaporation, or freeze-drying in the case of an aqueous solution.
  • Screening and characterization of pharmaceutically acceptable salts, polymorphs, and / or solvates can be accomplished using a variety of techniques, including but not limited to thermal analysis, X-ray diffraction, spectroscopy, microscopic methods, elemental analysis.
  • Various spectroscopic techniques used include, but are not limited to, Raman, FTIR, UVIS, and NMR (liquid and solid state).
  • Various microscopy techniques include but are not limited to IR microscopy and Raman microscopy.
  • the application also provides a pharmaceutical composition
  • a pharmaceutical composition comprising at least one compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, pharmaceutically active metabolite or prodrug of the compound, and a pharmaceutically acceptable Carriers or excipients, and optionally other therapeutic agents.
  • the drug containing the compound of the present invention can be administered to a patient by at least one of injection, oral, inhalation, rectal, and transdermal administration.
  • Other therapeutic agents can be selected from the following drugs: immunosuppressive agents (eg tacrolimus, cyclosporin, rapamycin, methotrexate, cyclophosphamide, azathioprine, mercaptopurine, mycophenolate or FTY720), glucocorticoid drugs (such as prednisone, cortisone acetate, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, hydroxyprednisolone, beclomethasone , Fluhydrocortisone acetate, deoxycorticosterone acetate, aldosterone), non-steroidal anti-inflammatory drugs (such as salicylate, arylalkanoic acid, 2-arylpropionic acid, N-arylanthranilic acid, Xicams, corxixes, or thioanilides), allergic vaccines, antihistamines, antileukotrien
  • the other therapeutic agents mentioned may also be rapamycin (Rapamycin), crizotinib (Crizotinib), tamoxifen, raloxifene, anastrozole, exemestane, letrozole , Herceptin TM (trastuzumab), Gleevec TM (imatinib), taxol TM (paclitaxel), cyclophosphamide, lovastatin, Miele tetracycline (Minosine), cytarabine, 5-fluorouracil (5-FU), methotrexate (MTX), taxotere TM (docetaxel), Zoladex TM (goserelin), vincristine, vinblastine, nocodazole oxazole, teniposide, etoposide, GEMZAR (TM) (gemcitabine), epothilone (epothilone), the promise of this CD, camptothecin, da
  • the other therapeutic agent may be a cytokine such as G-CSF (granulocyte colony stimulating factor).
  • other therapeutic agents may be, for example, but not limited to, CMF (cyclophosphamide, methotrexate, and 5-fluorouracil), CAF (cyclophosphamide, adriamycin, and 5-fluorouracil), AC (subcutaneous Dririamycin and cyclophosphamide), FEC (5-fluorouracil, epirubicin and cyclophosphamide), ACT or ATC (adriamycin, cyclophosphamide and paclitaxel) or CMFP (cyclophosphamide, (Aminopterin, 5-fluorouracil and prednisone).
  • CMF cyclophosphamide, methotrexate, and 5-fluorouracil
  • CAF cyclophosphamide, adriamycin, and 5-fluorouracil
  • the amount of a given drug depends on many factors, such as the specific dosing regimen, the type of disease or disorder and its severity, the subject in need of treatment Or the uniqueness of the host (e.g. body weight), however, depending on the specific surrounding conditions, including, for example, the specific drug that has been used, the route of administration, the condition being treated, and the subject or host being treated, the dosage administered may be known in the art
  • the method is routinely decided.
  • the dosage administered is typically in the range of 0.02-5000 mg / day, for example, about 1-1500 mg / day.
  • the desired dose may conveniently be presented as a single dose, or administered simultaneously (or within a short period of time) or in divided doses at appropriate intervals, such as two, three, four, or more divided doses per day.
  • dose range is given, the specific effective amount can be appropriately adjusted according to the condition of the patient and combined with the diagnosis of the physician.
  • the kinase inhibitors of the present invention include compounds of formula (I) or pharmaceutically acceptable salts, solvates, esters, acids, metabolites or prodrugs, or pharmaceutical compositions thereof for reducing or inhibiting the activity of cells or subjects cKIT (particularly mutant cKIT / T670I), FLT3 (including mutant FLT3-ITD), PDGFR ⁇ , PDGFR ⁇ , and / or VEGFR2 kinase activity, and / or prevent or treat cKIT (particularly mutant cKIT) in the subject / T670I), FLT3 (including mutant FLT3-ITD), PDGFR ⁇ , PDGFR ⁇ , and / or VEGFR2 activity related disorders.
  • cKIT particularly mutant cKIT / T670I
  • FLT3 including mutant FLT3-ITD
  • the compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug thereof, or a pharmaceutical composition thereof can be used to treat, prevent, or ameliorate one or more diseases selected from the group consisting of : Solid tumors (including benign or especially malignant types), especially sarcomas, gastrointestinal stromal tumors (Gastrointestinal Stromal Tumors, GIST), colorectal cancer (colon cancer), acute myeloid leukemia (Acute Myeloblastic Leukemia, AML), chronic Chronic Myelogenous Leukemia (CML), neoplasia, thyroid cancer, systemic mastocytosis, eosinophilia syndrome, fibrosis, lupus erythematosus, graft-versus-host disease, neurofibromatosis, pulmonary hypertension, Alzheimer's disease, seminoma, dysgerminoma, mast cell tumor, lung cancer, bronchial carcinoma, testicular intrae
  • gastrointestinal stromal tumors colorectal cancer, acute myeloid leukemia, chronic myeloid leukemia, thyroid cancer or similar diseases, or a combination thereof.
  • the inhibitor of the present invention or a pharmaceutical composition thereof can be used for the treatment or prevention of gastrointestinal stromal tumors, especially cKIT-T670I mutant gastrointestinal stromal tumors.
  • the compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug thereof, or a pharmaceutical composition thereof can be used to treat, prevent, or ameliorate an autoimmune disease selected from the group consisting of: Arthritis, rheumatoid arthritis, osteoarthritis, lupus, rheumatoid arthritis, inflammatory bowel disease, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes , Myasthenia gravis, Hashimoto's thyroiditis, Ord's hyroiditis, Graves' disease, rheumatoid arthritis syndrome ( syndrome), multiple sclerosis, infectious neuronitis (Guillain-Barrésyndrome), acute disseminated encephalomyelitis, Addison's disease, visual ocular twin-myoclonic syndrome, ankylosing Spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepati
  • the reactions can be used in order to provide the compounds described herein; or they can be used to synthesize fragments that are subsequently added by methods described herein and / or methods known in the art.
  • kinase inhibitor compounds described herein are provided herein and methods of use thereof.
  • the compounds described herein can be synthesized using the following synthetic schemes. Compounds can be synthesized by using appropriate alternative starting materials using methods similar to those described below.
  • the starting materials used to synthesize the compounds described herein can be synthesized or can be obtained from commercial sources.
  • the compounds described herein and other related compounds with different substituents can be synthesized using techniques and starting materials known to those skilled in the art.
  • the general method for preparing the compounds disclosed herein can be derived from reactions known in the art, and the reaction can be modified by reagents and conditions deemed appropriate by those skilled in the art to introduce various parts of the molecules provided herein.
  • reaction product can be isolated and purified using conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, and other methods. These products can be characterized using conventional methods, including physical constants and spectral data.
  • Non-limiting examples of synthetic schemes for preparing compounds of formula (I) are shown in the following synthetic routes.
  • N- (4-methylthiazol-2-yl) acetamide Add 4-methylthiazol-2-amine (2g) to a 100mL round bottom flask, add anhydrous dichloromethane (50mL), triethylamine ( 3.9mL), slowly add acetyl chloride (1.5mL) dropwise. The reaction system was reacted under argon protection at room temperature for 4 hours. After the reaction was completed, the system was evaporated to dryness under reduced pressure, and the resultant was neutralized with saturated sodium bicarbonate to pH> 10, and then extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate.
  • the reaction system was heated to 130 ° C for 14 hours under the protection of argon. After the reaction was completed, the system was evaporated to dryness under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. The organic phase was filtered and evaporated to dryness under reduced pressure to obtain a crude product. The crude product was purified by pressurized silica gel column chromatography to obtain pure product, MS (ESI) m / z (M + 1) +: 460.18.
  • the reaction system was stirred at room temperature for 14 hours under the protection of argon. After the reaction was completed, the system was evaporated to dryness under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. The organic phase was filtered and evaporated to dryness under reduced pressure to obtain a crude product.
  • the crude product was dissolved in anhydrous dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added.
  • the reaction system was stirred at room temperature for 14 hours under the protection of argon. After the reaction was completed, the system was evaporated to dryness under reduced pressure, and the resultant was diluted with water and neutralized with saturated sodium bicarbonate solution to pH> 10.
  • the aqueous phase was extracted with ethyl acetate, and the organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate.
  • the organic phase was filtered and evaporated to dryness under reduced pressure to obtain a crude product.
  • the crude product was purified by pressurized silica gel column chromatography to obtain compound 1, MS (ESI) m / z (M + 1) +: 550.24.
  • Example 2 The synthesis of Example 2 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 540.09.
  • Example 3 The synthesis of Example 3 was completed by using a procedure similar to that described in Example 1. MS (ESI) m / z (M + 1) +: 520.14.
  • Example 4 The synthesis of Example 4 was completed by using a procedure similar to that described in Example 1. MS (ESI) m / z (M + 1) +: 439.14.
  • Example 5 The synthesis of Example 5 was completed by using a procedure similar to that described in Example 1. MS (ESI) m / z (M + 1) +: 361.11.
  • Example 6 The synthesis of Example 6 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 506.13.
  • Example 7 The synthesis of Example 7 was completed using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 502.17.
  • Example 8 The synthesis of Example 8 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 512.18.
  • Example 9 The synthesis of Example 9 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 474.21.
  • Example 10 The synthesis of Example 10 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 486.12.
  • Example 11 The synthesis of Example 11 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 459.20.
  • Example 12 The synthesis of Example 12 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 419.17.
  • Example 13 The synthesis of Example 13 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 459.20.
  • Example 14 The synthesis of Example 14 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 445.18.
  • Example 15 The synthesis of Example 15 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 465.16.
  • Example 16 The synthesis of Example 16 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 458.20.
  • Example 17 The synthesis of Example 17 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 405.15.
  • Example 18 The synthesis of Example 18 was completed by using a procedure similar to that described in Example 1. MS (ESI) m / z (M + 1) +: 447.20.
  • Example 19 The synthesis of Example 19 was completed by using a procedure similar to that described in Example 1. MS (ESI) m / z (M + 1) +: 404.16.
  • Example 20 The synthesis of Example 20 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 481.18.
  • the reaction system was heated to 130 ° C for 14 hours under the protection of argon. After the reaction was completed, the system was evaporated to dryness under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. The organic phase was filtered and evaporated to dryness under reduced pressure to obtain a crude product. The crude product was purified by pressurized silica gel column chromatography to obtain pure product, MS (ESI) m / z (M + 1) +: 455.23.
  • N- (5- (3-iodo-1H-indazole-6-yl) -4-methylthiazol-2-yl) -2- (4-methylpiperazin-1-yl) acetamide (21d) Add N- (5- (1H-indazol-6-yl) -4-methylthiazol-2-yl) -2- (4-methylpiperazin-1-yl) ethane to a 50mL round bottom flask After amide (0.6g), N, N-dimethylformamide (10mL), iodine (0.8g) and potassium hydroxide (0.4g) were added. The reaction system was stirred at room temperature for 8 hours under the protection of argon.
  • the reaction system was heated to 80 ° C under the protection of argon for 14 hours. After the reaction was completed, the system was evaporated to dryness under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. The organic phase was filtered and evaporated to dryness under reduced pressure to obtain a crude product. The crude product was purified by pressurized silica gel column chromatography to obtain compound 21, MS (ESI) m / z (M + 1) +: 451.21.
  • Example 22 The synthesis of Example 22 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 448.18.
  • Example 23 The synthesis of Example 23 was completed by using a procedure similar to that described in Example 21. MS (ESI) m / z (M + 1) +: 464.20.
  • Example 24 The synthesis of Example 24 was completed by using a procedure similar to that described in Example 21. MS (ESI) m / z (M + 1) +: 448.19.
  • Example 25 The synthesis of Example 25 was completed by using a procedure similar to that described in Example 21. MS (ESI) m / z (M + 1) +: 466.18.
  • Example 26 The synthesis of Example 26 was completed by using a procedure similar to that described in Example 21. MS (ESI) m / z (M + 1) +: 465.19.
  • Example 27 The synthesis of Example 27 was completed by using a procedure similar to that described in Example 1. MS (ESI) m / z (M + 1) +: 473.21.
  • Example 28 The synthesis of Example 28 was completed by using a procedure similar to that described in Example 21. MS (ESI) m / z (M + 1) +: 494.13.
  • Example 29 The synthesis of Example 29 was completed using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 521.14.
  • Example 30 The synthesis of Example 30 was completed by using procedures similar to those described in Example 1. MS (ESI) m / z (M + 1) +: 376.13.
  • the human gastrointestinal stromal tumor cell line GIST-T1 (expressing wild-type C-KIT gene) (purchased from Cosmo Bio Co., Ltd. (Japan)
  • human gastrointestinal stromal tumor Cell line cells GIST-T1-T670I (expressing C-KIT-T670I mutant gene) (constructed by our laboratory using CRISPR technology)
  • mouse proto-B cell BaF3 (purchased from ATCC).
  • mouse Tel-cKit-BaF3 stable expression of C-KIT wild-type kinase
  • mouse Tel-cKit / T670I-BaF3 stable expression of cKIT T670I mutant kinase
  • mouse Tel-PDGFR ⁇ were also selected.
  • the construction method was: PCR was used to amplify the human C-KIT, C-KIT T670I, PDGFR ⁇ , PDGFR ⁇ , VEGFR2, FLT3 kinase region sequences, and inserted into the N-terminal TEL fragment and / Or NPM fragment and / or TPR fragment of MSCV-Puro vector (purchased from Clontech), stably transferred into mouse BaF3 cells by retrovirus method, and the IL-3 growth factor was removed, and finally C-KIT, C-dependent -KIT / T670I, PDGFR ⁇ , PDGFR ⁇ , VEGFR2, FLT3 transfected protein cell lines.
  • the GIST-T1-T670I (expressing the C-KIT-T670I mutant gene) cell line was constructed by our laboratory.
  • the construction method was to target the sgRNA near the T670 site of the KIT gene by the CRISPR design tool of the Zhang Feng laboratory of the Massachusetts Institute of Technology Designed (website: crispr.mit.edu), and cloned into pSpCas9 (BB) -2A-Puro vector (Addgene, USA); the obtained vector and a segment of single-stranded oligomer with T670I mutation near T670 site Cells were co-transfected with nucleotides, after antibiotic selection, diluted and single-cell cultured in 96-well plates; Sanger sequencing method was used to verify the T670 site of cells.
  • the compounds of the present invention and the control compound axiti of different concentrations (0.000508 ⁇ M, 0.00152 ⁇ M, 0.00457 ⁇ M, 0.0137 ⁇ M, 0.0411 ⁇ M, 0.123 ⁇ M, 0.370 ⁇ M, 1.11 ⁇ M, 3.33 ⁇ M, 10 ⁇ M in DMSO)
  • Axitinib purchased from China, MedChem Express
  • CCK-8 purchased from Bebo Biotech Co., Shanghai, China
  • CCK-8 cell viability detection kit
  • the dehydrogenase in is reduced to a yellow formazan product with high water solubility.
  • the amount of formazan produced is proportional to the number of live cells.
  • the experimental results shown in Table 2 show that the compounds of the present invention have a certain inhibitory effect on mutant cKIT-T670I, VEGFR2, PDGFR ⁇ , PDGFR ⁇ , FLT3, especially compared with cKIT wild type, it has more inhibitory effect on mutant cKIT-T670I Strong inhibitory effect.
  • the preferred compounds of the present invention have comparable or stronger inhibitory activity against mutant cKIT-T670I, but relatively weaker inhibitory activity against wild-type cKIT.
  • cKIT wild type plays a very important role in the early development of normal hematopoietic stem cells, so under unnecessary circumstances, inhibition of cKIT kinase will produce mechanistic toxicity, and there are reports in the literature that the simultaneous inhibition of FLT3 and cKIT will cause bone marrow suppression toxicity .
  • Axitinib has a certain inhibitory effect on maternal BaF3 cells, and there is no selectivity between wild-type cKIT and mutant cKIT-T670I; and the compounds of the present invention are between mutant cKIT-T670I and wild-type cKIT and maternal BaF3
  • the cells showed obvious selective inhibition, indicating that the preferred compounds of the present invention, while inhibiting mutant cKIT-T670I, will not cause the problem of myelosuppressive toxicity due to the inhibition of wild-type cKIT and FLT3.
  • the compounds 7 and 9 of the present invention and the control compound Imatinib (purchased from China, MedChem Express) were resistant to the gastrointestinal stromal tumor cell line GIST-T1 and the Imaitinib constructed by our laboratory Testing on the mutated GIST-T1-T670I cell line revealed that the compounds of the present invention not only have a strong inhibitory effect on gastrointestinal stromal tumors sensitive to Imatinib, but also have a strong inhibition on GIST-T1-T670I resistant to Imatinib effect. This indicates that the compounds of the present invention can be used to treat T670I mutant gastrointestinal stromal tumors.
  • mice were orally administered methylcellulose (HKI) vehicle (5 mice) daily; the dose was 10 mg / kg , 20 mg / kg, 40 mg / kg, 100 mg / kg compound 9 (5 mice each); dose of 40 mg / kg mouse weight Sunitinib (purchased from MedChemExpress, China) (5 mice) ).
  • HKI methylcellulose
  • mice For the mouse model of GIST-T1-T670I, starting from the 15th day, the corresponding mice were orally administered methylcellulose (HKI) vehicle (5 mice); the doses were 20 mg / kg, 30 mg / kg , 40 mg / kg of compound 9 (5 mice each); sunitinib (5 mice) at a dose of 40 mg / kg mouse weight.
  • HKI methylcellulose
  • Compound 9 has shown a certain effect of inhibiting tumors in mice in the TEL-cKIT / T670I-BaF3 and GIST-T1-T670I mouse tumor models at a dosage of 40 mg / kg, and with the increase in the number of days of medication, The inhibitory effect of compound 9 on mouse tumors is more significant, and the tumor inhibition rate is as high as 80% or more.
  • the tumor inhibition is 11 days after administration The rate reached 100%.
  • the tumor inhibition rate on the 28th day after administration in the GIST-T1-T670I mouse model was 84.3%.
  • Compound 9 not only effectively inhibited the growth of tumors in mice, but basically had no effect on the body weight of mice, indicating that compound 9 can be applied to animal administration. This also proves that the CKIT / T670I inhibitor compound of the present invention can be used to treat gastrointestinal stromal tumors with T670I mutation.
  • the present invention provides a novel kinase inhibitor compound that can be used to reduce or inhibit cKIT (especially mutant cKIT / T670I), FLT3 (including mutant FLT3-ITD), PDGFR ⁇ , PDGFR ⁇ , and / Or VEGFR2 kinase activity, and / or prevent or treat cKIT (especially mutant cKIT / T670I), FLT3 (including mutant FLT3-ITD), PDGFR ⁇ , PDGFR ⁇ , and / or VEGFR2 activity related disorders in the subject. Therefore, it can be made into a corresponding medicine, which is suitable for industrial applications.
  • the present invention provides a novel kinase inhibitor compound that can be used to reduce or inhibit cKIT (especially mutant cKIT / T670I), FLT3 (including mutant FLT3-ITD), PDGFR ⁇ , PDGFR ⁇ , and / Or VEGFR2 kinase activity, and / or prevent or treat cKIT (especially mutant cKIT / T670I), FLT3 (including mutant FLT3-ITD), PDGFR ⁇ , PDGFR ⁇ , and / or VEGFR2 activity related disorders in the subject. Therefore, it can be made into a corresponding medicine, which is suitable for industrial applications.

Abstract

本发明涉及一种激酶抑制剂，其包括式(I)的化合物或其药学可接受的盐、溶剂化物、酯、酸、代谢物或前药。本发明还涉及包括该激酶抑制剂的药物组合物，以及使用这些化合物和组合物来抑制细胞或受试者的cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2激酶活性，并且/或者在受试者中预防或治疗cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2活性相关病症的用途和方法。(I)

Description

吲唑类激酶抑制剂及其用途 技术领域
本申请涉及一种激酶抑制剂、以及使用这样的激酶抑制剂抑制激酶活性的方法和用途。更具体地，本发明涉及能够抑制cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2激酶活性的抑制剂。
背景技术
胃肠间质瘤(gastrointestinal stromal tumors，GIST)是胃肠道最常见的间叶源性肿瘤。GIST的发病率约为1/10万～2/10万，占全部消化道肿瘤的1-3％。该病多见于中老年人，发病的中位年龄为50～65岁，40岁以前少见，但在儿童中也有报道。目前认为GIST是一种具有潜在恶性行为的肿瘤，其生物学行为难以预测。GIST可发生于消化道任何部位，最为常见的是胃(60％～70％)，其次为小肠(20％～30％)，在食管、结肠、直肠发生率不足10％，也可发生于网膜和肠系膜。
根据临床研究，胃肠间质瘤的发病机制根据其基因分子分型可分为三类：cKIT突变型(80-85％)、PDGFRα突变型(5-10％)和cKIT野生型GISTs(10％)。胃肠间质瘤的发病机制与cKIT蛋白(CD117)信号通路的激活有关。原癌基因cKIT是从猫科纤维肉瘤病毒中分离出的vKIT基因的同源物，它位于人体4号染色体(4q12-13)上，长约90kb，由21个外显子、20个内显子组成，在进化过程中高度保守。cKIT蛋白是一种位于细胞膜上的受体酪氨酸激酶(Receptor Tyrosine Kinase，RTK)，相对分子质量为145000，按其细胞表面抗原决定簇被命名为CD117。cKIT蛋白属于第三型RTK家族，由含有5个免疫球蛋白样结构域(D1～D5)、1个跨膜结构域以及1个含有近膜结构域(JMD)和酪氨酸激酶(TK)结构域的胞质区构成。TK结构域又分为三磷酸腺苷(adenosine triphosphate，ATP)结构域(TK1)和磷酸转移酶(phosphotransferase)结构域(TK2)。配体干细胞因子SCF(stem cell factor)与胞外结构域结合形成二聚体，导致胞质区TK结构域内酪氨酸自身磷酸化，进一步引起下游各种效应的自磷酸化，完成各种信 号的传递。主要的信号通路有PI3K信号通路、JAK-STAT信号通路、Ras-Erk信号通路、Src家族激酶信号通路以及PLC信号通路等，最终促进细胞的增生、***以及组织的生长、存活。
目前，手术作为传统的外科治疗手段仍是治疗胃肠间质瘤最主要的方法，而近年来兴起的靶向药物则开启了治疗GIST的一个新的阶段。到目前为止临床上用于治疗GIST的cKIT激酶抑制剂主要有诺华公司的伊马替尼Imatinib(cKIT/BCR-ABL/PDGFR)和辉瑞公司的舒尼替尼Sunitinib(cKIT/BCR-ABL/PDGFR/VEGFR2/FLT3)。Imatinib是第一个用于治疗GIST的II型激酶抑制剂，Sunitinib是于2006年获得FDA批准的I型cKIT激酶抑制剂。此外具有cKIT活性的II型激酶抑制剂还有瑞格菲尼Regorafenib、尼罗替尼Nilotinib、马赛替尼Mastinib、索拉非尼Sorafenib等。这些小分子抑制剂均为多靶点抑制剂，其中Nilotinib和Mastinib不能克服T670I突变。拜耳公司研发的两个具有cKIT活性的小分子抑制剂Regorafenib和Sorafenib虽然对cKIT-T670I突变具有一定活性，但是二者均为多靶点抑制剂，除了对cKIT有作用，还对FLT3/PDGFR/FGFR/VEGFR2/RET/RAF等多种激酶有较强的作用。阿西替尼Axitinib是由辉瑞公司研发的多靶点小分子抑制剂，同时具有cKIT、BCR-ABL、VEGFR2等激酶的抑制活性。此外，普纳替尼Ponatinib也有对cKIT激酶也有很强的抑制活性，同时它还可以克服cKIT-T670I突变产生的耐药，但它是ABL/PDGFR/RET/CSF1R/FGFR/VEGFR/FGFR/RET等多种激酶的多靶点抑制剂。
本发明主要是发现一种结构全新的针对Imatinib耐药突变cKIT-T670I有很强抑制作用的抑制剂。
发明内容
本发明提供一种选择性的激酶抑制剂，包括式(I)的化合物或其药学可接受的盐、溶剂化物、酯、酸、代谢物或前药：
Figure PCTCN2018115025-appb-000001
其中，
X为-(CH＝CH) m-，其中m为0或1；
Y选自-NH-或-(CH 2) n-，其中n为0-3的整数；
R 1选自任选地被1-3个独立的R 4基团取代的苯基、任选地被1-3个独立的R 4基团取代的吡啶基、任选地被1-3个独立的R 4基团取代的吡唑基、和任选地被1-3个独立的R 4基团取代的嘧啶基；
R 2选自氢和C 1-6烷基；
R 3选自任选地被1-2个独立的R 5基团取代的C 1-6烷基、C 1-6烷基氨基、以及任选地被1-3个独立的R 4基团取代的苯基、任选地被1-3个独立的R 4基团取代的萘基、任选地被1-3个独立的R 4基团取代的吡啶基、任选地被1-3个独立的R 4基团取代的哌嗪基、和任选地被1-3个独立的R 4基团取代的哌啶基；
R 4独立地选自卤素、氨基、C 1-6烷基、C 3-6环烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6羟基烷基、C 1-6烷基氨基、C 2-6烷酰胺基、(4-甲基哌嗪-1-基)甲基、吗啉甲基、吗啉基、4-甲基哌嗪-1-基、4-哌啶基、和4-四氢吡喃基；
R 5独立地选自氨基、羟基、和C 1-6烷硫基。
在优选的实施方式中，X为-(CH＝CH)-。
在另一优选的实施方式中，Y为直接键或-CH 2-。
在另外优选的实施方式中，R 1选自任选地被1-3个独立的R 4基团取代的苯基、吡啶基、吡唑基、和嘧啶基，其中R 4独立地选自卤素、氨基、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基、和(4-甲基哌嗪-1-基)甲基；R 1更优选为任选地被甲基、氨基或卤素取代的苯基、2-吡啶基、3-吡啶基、4-吡唑基、和5-嘧啶基；R 1特别优选为2-吡啶基。
在另外优选的实施方式中，R 2为氢或甲基。
在另外优选的实施方式中，R 3选自任选地被1-2个独立的R 5基团 取代的C 1-6烷基、C 1-6烷基氨基、以及任选地被1-3个独立的R 4基团取代的苯基、萘基、吡啶基、哌嗪基、和哌啶基，其中R 4独立地选自卤素、氨基、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基、和(4-甲基哌嗪-1-基)甲基，R 5独立地选自氨基、羟基、和甲硫基；R 3更优选为任选地被氨基、羟基、或甲硫基取代的C 1-6烷基、二甲氨基、任选地被甲基取代的N-哌嗪基、任选地被卤素、三氟甲基、或甲氧基取代的苯基、萘基、4-吡啶基、3-哌啶基、和任选地被甲基取代的4-哌啶基；R 3特别优选为甲基、2-丙基、1-氨基-3-甲硫基-丙基、1-氨基-3-甲基-丁基、甲基取代的N-哌嗪基、甲氧基取代的苯基、4-吡啶基、或4-哌啶基。
在特别优选的实施方式中，当Y为直接键时，R 3选自任选地被氨基、羟基、或甲硫基取代的C 1-6烷基、和4-吡啶基；当Y为-CH 2-时，R 3选自任选地被甲氧基取代的苯基、任选地被甲基取代的N-哌嗪基、和任选地被甲基取代的4-哌啶基。
在另一方面，本发明还涉及一种药物组合物，其包括本发明的激酶抑制剂，以及药学上可接受的载体或赋形剂，以及任选的其它治疗剂。
在其他方面，本发明还涉及采用激酶抑制剂或包括其的药物组合物降低或抑制细胞或受试者的cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2激酶活性的方法和用途。
在又一方面，本发明还涉及采用激酶抑制剂或包括其的药物组合物在受试者中预防或治疗cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2活性相关病症的方法和用途。
附图说明
图1a示出化合物9和舒尼替尼在tel-cKIT/T670I-BaF3细胞肿瘤移植小鼠模型中给药后对小鼠体重的影响；图1b示出化合物9和舒尼替尼在tel-cKIT/T670I-BaF3细胞肿瘤移植小鼠模型中的肿瘤抑制效果。
图2a示出化合物9和舒尼替尼在GIST-T1-T670I细胞肿瘤移植小鼠模型中给药后对小鼠体重的影响；图2b示出化合物9和舒尼替尼在GIST-T1-T670I细胞肿瘤移植小鼠模型中的肿瘤抑制效果。
具体实施方式
术语
除非另外定义，所有本文使用的科技术语都具有与要求保护的主题所属领域的技术人员一般理解相同的含义。
除非另有说明，本发明采用本领域技术范围内的质谱、NMR、HPLC、蛋白质化学、生物化学、重组DNA技术和药理学等常规方法。除非提供具体的定义，否则与本文描述的分析化学、合成有机化学、以及医学和药物化学等化学上相关的命名和实验室操作和技术，是本领域技术人员已知的。一般而言，前述技术和步骤可以通过本领域众所周知的和在各种一般文献和更具体文献中描述的常规方法来实施，这些文献在本说明书中被引用和讨论。
术语“烷基”是指脂肪族烃基团，可以是支链或直链的烷基。根据结构，烷基可以是单价基团或双价基团(即亚烷基)。在本发明中，烷基优选是具有1-8个碳原子的烷基，更优选具有1-6个碳原子的“低级烷基”，甚至更优选具有1-4个碳原子的烷基。典型的烷基包括但不限于甲基、乙基、丙基、丁基、戊基、己基等。应理解，本文提到的“烷基”包括可能存在的所有构型和构象的该烷基，例如本文提到的“丙基”包括正丙基和异丙基，“丁基”包括正丁基、异丁基和叔丁基，“戊基”包括正戊基、异丙基、新戊基、叔戊基、和戊-3-基等。
术语“烷氧基”是指-O-烷基，其中烷基如本文中定义。典型的烷氧基包括但不限于甲氧基、乙氧基、丙氧基、丁氧基、戊氧基、己氧基等。
术语“烷氧基烷基”是指本文定义的烷基被本文定义的烷氧基取代。
术语“环烷基”是指单环或多环基，其仅含有碳和氢。环烷基包括具有3-12个环原子的基团。根据结构，环烷基可以是单价基团或双价基团(例如亚环烷基)。在本发明中，环烷基优选是具有3-8个碳原子 的环烷基，更优选具有3-6个碳原子的“低级环烷基”。环烷基的例子包括但不限于，环丙基、环丁基、环戊基、环己基、环庚基、环辛基、环戊烯基、环己烯基、环庚烯基和金刚烷基。
术语“烷基(环烷基)”或“环烷基烷基”是指本文定义的烷基被本文定义的环烷基取代。非限制性的环烷基烷基包括环丙基甲基、环丁基甲基、环戊基甲基、环己基甲基等。
术语“芳香基”是指平面环具有离域的π电子***并且含有4n+2个π电子，其中n是整数。芳香基环可以由五、六、七、八、九或多于九个原子构成。芳香基可以是任选取代的。术语“芳香基”包括碳环芳基(例如苯基)和杂环芳基(或“杂芳基”或“杂芳香基”)基团(例如吡啶)。该术语包括单环或稠环多环(即共用相邻的碳原子对的环)基团。
本文使用的术语“芳基”是指芳香基环中每一个构成环的原子都是碳原子。芳基环可以由五、六、七、八、九或多于九个原子构成。芳基可以是任选取代的。芳基的实例包括但不限于苯基、萘基、菲基、蒽基、芴基和茚基。根据结构，芳基可以是单价基团或双价基团(即亚芳基)。
术语“芳氧基”是指-O-芳基，其中芳基如本文中定义。
术语“杂芳基”是指芳基中包括一个或多个选自氮、氧和硫的环杂原子。含N“杂芳基”部分是指芳香基中环上至少有一个骨架原子是氮原子。根据结构，杂芳基可以是单价基团或双价基团(即亚杂芳基)。杂芳基的实例包括但不限于吡啶基、咪唑基、嘧啶基、吡唑基、***基、吡嗪基、四唑基、呋喃基、噻吩基、异噁唑基、噻唑基、噁唑基、异噻唑基、吡咯基、喹啉基、异喹啉基、吲哚基、苯并咪唑基、苯并呋喃基、吲唑基、吲嗪基、酞嗪基、哒嗪基、异吲哚基、蝶啶基、嘌呤基、噁二唑基、噻二唑基、呋咱基、苯并呋咱基、苯并噻吩基、苯并噻唑基、苯并噁唑基、喹唑啉基、萘啶基和呋喃并吡啶基等。
术语“烷基(芳基)”或“芳烷基”是指本文定义的烷基被本文定义的芳基取代。非限制性的烷基(芳基)包括苄基、苯乙基等。
术语“烷基(杂芳基)”或“杂芳基烷基”是指本文定义的烷基被本文定义的杂芳基取代。
本文使用的术语“杂烷基”是指本文定义的烷基中的一个或多个骨 架链原子是杂原子，例如氧、氮、硫、硅、磷或它们的组合。所述杂原子(一个或多个)可以位于杂烷基内部的任意位置或在杂烷基与分子的其余部分相连的位置。
本文使用的术语“杂环烷基”或“杂环基”是指非芳香基环中一个或多个构成环的原子是选自氮、氧和硫的杂原子。杂环烷基环可以由三、四、五、六、七、八、九或多于九个原子构成。杂环烷基环可以是任选取代的。杂环烷基的实例包括但不限于内酰胺、内酯、环亚胺、环硫代亚胺、环氨基甲酸酯、四氢噻喃、4H-吡喃、四氢吡喃、哌啶、1,3-二噁英、1,3-二噁烷、1,4-二噁英、1,4-二噁烷、哌嗪、1,3-氧硫杂环己烷、1,4-氧硫杂环己二烯、1,4-氧硫杂环己烷、四氢-1,4-噻嗪、2H-1,2-噁嗪、马来酰亚胺、琥珀酰亚胺、巴比妥酸、硫代巴比妥酸、二氧代哌嗪、乙内酰脲、二氢尿嘧啶、吗啉、三噁烷、六氢-1,3,5-三嗪、四氢噻吩、四氢呋喃、吡咯啉、吡咯烷、咪唑烷，吡咯烷酮、吡唑啉、吡唑烷、咪唑啉、咪唑烷、1,3-二氧杂环戊烯、1,3-二氧杂环戊烷、1,3-二硫杂环戊烯、1,3-二硫杂环戊烷、异噁唑啉、异噁唑烷、噁唑啉、噁唑烷、噁唑烷酮、噻唑啉、噻唑烷和1,3-氧硫杂环戊烷。根据结构，杂环烷基可以是单价基团或双价基团(即亚杂环烷基)。
术语“烷基(杂环烷基)”或“杂环烷基烷基”是指本文定义的烷基被本文定义的杂环烷基取代。
术语“烷氧基(杂环烷基)”或“杂环烷基烷氧基”是指本文定义的烷氧基被本文定义的杂环烷基取代。
术语“卤”或“卤素”是指氟、氯、溴和碘。
术语“卤代烷基”、“卤代烷氧基”和“卤代杂烷基”包括烷基、烷氧基或杂烷基的结构，其中至少一个氢被卤原子置换。在某些实施方式中，如果两个或更多氢原子被卤原子置换，所述卤原子彼此相同或不同。
术语“羟基”是指-OH基团。
术语“氰基”是指-CN基团。
术语“酯基”是指具有式-COOR的化学部分，其中R选自烷基、环烷基、芳基、杂芳基(通过环碳连接)和杂环基(通过环碳连接)。
术语“氨基”是指-NH 2基团。
术语“氨酰基”是指-CO-NH 2基团。
术语“酰胺基”或“酰氨基”是指-NR-CO-R’，其中R和R’各自独立地为氢或烷基。
术语“烷基氨基”是指进一步被一个或两个烷基取代的氨基取代基，具体是指基团-NRR’，其中R和R’各自独立地选自氢或低级烷基，条件是-NRR’不是-NH 2。“烷基氨基”包括其中-NH 2的氮连接至少一个烷基基团的化合物的基团。烷基氨基基团的例子包括但不限于，甲基氨基、乙基氨基等。“二烷基氨基”包括其中-NH 2的氮连接至少两个其它烷基基团的基团。二烷基氨基基团的例子包括但不限于，二甲基氨基、二乙基氨基等。
术语“芳基氨基”和“二芳基氨基”是指进一步被一个或两个芳基取代的氨基取代基，具体是指基团-NRR’，其中R和R’各自独立地选自氢、低级烷基、或芳基，其中N分别连接至少一个或两个芳基基团。
术语“环烷基氨基”是指进一步被一个或两个本文所定义的环烷基取代的氨基取代基。
术语“杂烷基氨基”是指进一步被一个或两个本文所定义的杂烷基取代的氨基取代基。
本文的术语“芳烷基氨基”是指其中R是低级芳烷基且R’是氢、低级烷基、芳基或低级芳烷基的基团-NRR’。
术语“杂芳基氨基”是指进一步被一个或两个本文所定义的杂芳基取代的氨基取代基。
术语“杂环烷基氨基”是指本文定义的氨基被本文定义的杂环烷基取代。
术语“烷基氨基烷基”是指本文定义的烷基被本文定义的烷基氨基取代。
术语“氨基烷基”是指进一步被一个或多个氨基取代的烷基取代基。
术语“氨基烷氧基”是指进一步被一个或多个氨基取代的烷氧基取代基。
术语“羟烷基”或“羟基烷基”是指进一步被一个或多个羟基取代的烷基取代基。
术语“氰基烷基”是指进一步被一个或多个氰基取代的烷基取代 基。
术语“酰基”是指有机或无机含氧酸去掉羟基后剩下的一价原子团，通式为R-M(O)-，其中M通常为C。
术语“羰基”是由碳和氧两种原子通过双键连接而成的有机官能团(C＝O)。
术语“烷酰基”或“烷基羰基”是指进一步被一个烷基取代的羰基。典型的烷酰基包括但不限于乙酰基、丙酰基、丁酰基、戊酰基、己酰基等。
术语“芳基羰基”是指本文定义的羰基被本文定义的芳基取代。
术语“烷氧基羰基”是指进一步被一个烷氧基取代的羰基。
术语“杂环烷基羰基”是指进一步被一个杂环烷基取代的羰基。
术语“烷基氨基羰基”、“环烷基氨基羰基”、“芳基氨基羰基”、“芳烷基氨基羰基”、“杂芳基氨基羰基”分别是指本文定义的羰基分别被本文定义的烷基氨基、环烷基氨基、芳基氨基、芳烷基氨基、或杂芳基氨基取代。
术语“烷基羰基烷基”或“烷酰基烷基”是指进一步被一个烷基羰基取代的烷基。
术语“烷基羰基烷氧基”或“烷酰基烷氧基”是指进一步被一个烷基羰基取代的烷氧基。
术语“杂环烷基羰基烷基”是指进一步被一个杂环烷基羰基取代的烷基。
术语“巯基”是指-SH基团。术语“烷硫基”是指本文所定义的巯基被本文所定义的烷基取代。
术语“砜基”或“磺酰基”是指磺酸失去羟基后的官能团，具体是指-S(＝O) 2-基团。
术语“亚砜基”或“亚磺酰基”是指-S(＝O)-。
术语“氨基砜基”或“氨基磺酰基”是指-S(＝O) 2-NH 2基团。
术语“烷基亚砜基”或“烷基亚磺酰基”是指-S(＝O)-R，其中R为烷基。
术语“烷基砜基”或“烷基磺酰基”是指-S(＝O) 2-R，其中R为烷基。
术语“烷基氨基砜基”是指本文定义的砜基被本文定义的烷基氨基 取代。
术语“烷基砜基氨基”或“环烷基砜基氨基”是指本文定义的氨基被本文定义的烷基砜基或环烷基砜基取代。
术语“环烷基砜基”和“环烷基磺酰基”是指-S(＝O) 2-R，其中R为环烷基。
术语“烷基磺酰胺基”和“环烷基磺酰胺基”是指-NH-S(＝O) 2-R，其中R分别为烷基和环烷基。
术语“季铵基”是指-N +RR’R”，其中R、R’和R”各自独立地选自具有1-8个碳原子的烷基。
术语“任选”指后面描述的一个或多个事件可以发生或可以不发生，并且包括发生的事件和不发生的事件两者。术语“任选取代的”或“取代的”是指所提及的基团可以被一个或多个额外的基团取代，所述额外的基团各自并且独立地选自烷基、环烷基、芳基、杂芳基、杂环基、羟基、烷氧基、氰基、卤素、酰胺基、硝基、卤代烷基、氨基、甲磺酰基、烷基羰基、烷氧基羰基、杂芳基烷基、杂环烷基烷基、氨酰基、氨基保护基等。其中，氨基保护基优选选自新戊酰基、叔丁氧羰基、苄氧羰基、9-芴甲氧羰基、苄基、对甲氧苄基、烯丙氧羰基、和三氟乙酰基等。
本文使用的术语“酪氨酸蛋白激酶(tyrosine protein kinase，TPK)”是一类催化ATP上γ-磷酸转移到蛋白酪氨酸残基上的激酶，能催化多种底物蛋白质酪氨酸残基磷酸化，在细胞生长、增殖、分化中具有重要作用。
本文使用的术语激酶的“抑制”、“抑制的”或“抑制剂”，是指磷酸转移酶活性被抑制。
本文公开的化合物的“代谢物”是当该化合物被代谢时形成的化合物的衍生物。术语“活性代谢物”是指当该化合物被代谢时形成的化合物的生物活性衍生物。本文使用的术语“被代谢”，是指特定物质被生物体改变的过程总和(包括但不限于水解反应和由酶催化的反应，例如氧化反应)。因此，酶可以产生特定的结构转变为化合物。例如，细胞色素P450催化各种氧化和还原反应，同时二磷酸葡萄糖甘酸基转移酶催化活化的葡萄糖醛酸分子至芳香醇、脂肪族醇、羧酸、胺和游离 的巯基的转化。新陈代谢的进一步的信息可以从《The Pharmacological Basis of Therapeutics》，第九版，McGraw-Hill(1996)获得。本文公开的化合物的代谢物可以通过将化合物给予宿主并分析来自该宿主的组织样品、或通过将化合物与肝细胞在体外孵育并且分析所得化合物来鉴别。这两种方法都是本领域已知的。在一些实施方式中，化合物的代谢物是通过氧化过程形成并与相应的含羟基化合物对应。在一些实施方式中，化合物被代谢为药物活性代谢物。本文使用的术语“调节”，是指直接或间接与靶标相互作用，以改变靶标的活性，仅仅举例来说，包括增强靶标的活性、抑制靶标的活性、限制靶标的活性或者延长靶标的活性。
本文使用的术语“靶蛋白”是指能被选择性结合化合物所结合的蛋白质分子或部分蛋白质。在某些实施方式中，靶蛋白是酪氨酸激酶KIT(野生型或各种突变或其组合)、ABL(野生型或各种突变或其组合)、EGFR(野生型或各种突变或其组合)、FLT3(野生型或各种突变或其组合)、VEGFR2(野生型或各种突变或其组合)、RET(野生型或各种突变或其组合)、PDGFRα(野生型或各种突变或其组合)、PDGFRβ(野生型或各种突变或其组合)、BCR/ABL(野生型或各种突变或其组合)、FGFR1(野生型或各种突变或其组合)、FGFR2(野生型或各种突变或其组合)、FGFR3(野生型或各种突变或其组合)、FGFR4(野生型或各种突变或其组合)。
本文使用的IC 50是指在测量这样的效应的分析中获得最大效应的50％抑制的特定测试化合物的量、浓度或剂量。
本文使用的EC 50是指测定化合物的剂量、浓度或量，其引起特定测定化合物诱导、刺激或加强的特定反应的50％的最大表达的剂量依赖反应。
本文使用的GI 50是指使50％细胞生长被抑制所需的药物浓度，即药物使50％细胞(如癌细胞)的生长得到抑制或控制时的药物浓度。
本发明新型的激酶抑制剂
本发明提供一种新型的激酶抑制剂，包括式(I)的化合物或其药学可接受的盐、溶剂化物、酯、酸、代谢物或前体药物，
Figure PCTCN2018115025-appb-000002
其中，
X为-(CH＝CH) m-，其中m为0或1，当m为0时X表示直接键；
Y选自-NH-或-(CH 2) n-，其中n为0-3的整数，当n为0时Y表示直接键；
R 1选自任选地被1-3个独立的R 4基团取代的芳基和杂芳基；
R 2选自氢和C 1-6烷基；
R 3选自任选地被1-2个独立的R 5基团取代的C 1-6烷基、C 1-6烷基氨基、以及任选地被1-3个独立的R 4基团取代的芳基、杂芳基和杂环基；
R 4独立地选自卤素、氨基、C 1-6烷基、C 3-6环烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6羟基烷基、C 1-6烷基氨基、C 2-6烷酰胺基、(4-甲基哌嗪-1-基)甲基、吗啉甲基、吗啉基、4-甲基哌嗪-1-基、4-哌啶基、和4-四氢吡喃基；
R 5独立地选自氨基、羟基、和C 1-6烷硫基。
在优选的实施方式中，X为-(CH＝CH)-。
在另一优选的实施方式中，Y为直接键或-CH 2-。
在另外优选的实施方式中，R 1选自任选地被1-3个独立的R 4基团取代的苯基、吡啶基、吡唑基、和嘧啶基，其中R 4独立地选自卤素、氨基、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基、和(4-甲基哌嗪-1-基)甲基；R 1更优选为任选地被甲基、氨基或卤素取代的苯基、2-吡啶基、3-吡啶基、4-吡唑基、和5-嘧啶基；R 1特别优选为2-吡啶基。
在另外优选的实施方式中，R 2为氢或甲基。
在另外优选的实施方式中，R 3选自任选地被1-2个独立的R 5基团取代的C 1-6烷基、C 1-6烷基氨基、以及任选地被1-3个独立的R 4基团取代的苯基、萘基、吡啶基、哌嗪基、和哌啶基，其中R 4独立地选自卤素、氨基、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基、和(4-甲基哌嗪-1-基) 甲基，R 5独立地选自氨基、羟基、和甲硫基；R 3更优选为任选地被氨基、羟基、或甲硫基取代的C 1-6烷基、二甲氨基、任选地被甲基取代的N-哌嗪基、任选地被卤素、三氟甲基、或甲氧基取代的苯基、萘基、4-吡啶基、3-哌啶基、和任选地被甲基取代的4-哌啶基；R 3特别优选为甲基、2-丙基、1-氨基-3-甲硫基-丙基、1-氨基-3-甲基-丁基、甲基取代的N-哌嗪基、甲氧基取代的苯基、4-吡啶基、或4-哌啶基。
在特别优选的实施方式中，当Y为直接键时，R 3选自任选地被氨基、羟基、或甲硫基取代的C 1-6烷基、和4-吡啶基；当Y为-CH 2-时，R 3选自任选地被甲氧基取代的苯基、任选地被甲基取代的N-哌嗪基、和任选地被甲基取代的4-哌啶基。
对于各个变量，上述基团的任意组合也在本文考虑之中。可以理解的是：本文所提供的化合物上的取代基和取代模式可以由本领域技术人员进行选择，以便提供化学上稳定的且可以使用本领域已知的技术以及本文阐述的技术合成的化合物。
在优选的实施方式中，本发明的抑制剂包括下表1的化合物或其药学可接受的盐、溶剂化物、酯、酸、代谢物或前药。
表1
Figure PCTCN2018115025-appb-000003
Figure PCTCN2018115025-appb-000004
Figure PCTCN2018115025-appb-000005
本文描述的是新型的激酶抑制剂。本文也描述了此化合物的药学可接受的盐、溶剂化物、酯、酸、药物活性代谢物和前药。
在另外的或进一步的实施方式中，将本文描述的化合物给予有需要的生物体后在其体内代谢产生代谢物，所产生的代谢物然后用于产生期望的效果，包括期望的治疗效果。
本文描述的化合物可以被制成和/或被用作药学可接受的盐。药学可接受的盐的类型包括但不限于：(1)酸加成盐、通过将化合物的游离碱形式与药学可接受的无机酸反应形成，所述无机酸如盐酸、氢溴酸、硫酸、硝酸、磷酸、偏磷酸等；或与有机酸反应形成，所述有机酸如乙酸、丙酸、己酸、环戊烷丙酸、羟基乙酸、丙酮酸、乳酸、丙二酸、苹果酸、柠檬酸、琥珀酸、马来酸、酒石酸、反丁烯二酸、三氟乙酸、苯甲酸、3-(4-羟基苯甲酰基)苯甲酸、肉桂酸、扁桃酸、甲烷磺酸、乙烷磺酸、1,2-乙二磺酸、2-羟基乙磺酸、苯磺酸、甲苯磺酸、4-甲基双环-[2.2.2]辛-2-烯-1-甲酸、2-萘磺酸、叔丁基乙酸、葡庚糖酸、4,4'-亚甲基双-(3-羟基-2-烯-1-甲酸)、3-苯基丙酸、三甲基乙酸、十二烷基硫酸、葡糖酸、谷氨酸、水杨酸、羟基萘酸、硬脂酸、粘康酸等；(2)碱加成盐，其在母体化合物中的酸性质子被金属离子置换时形成，例如碱金属离子(例如锂、钠、钾)、碱土金属离子(例如镁或钙)或铝离子；或与有机碱或无机碱配位，可接受的有机碱包括乙醇胺、二乙 醇胺、三乙醇胺、三甲胺、N-甲基葡萄糖胺等；可接受的无机碱包括氢氧化铝、氢氧化钙、氢氧化钾、碳酸钠、氢氧化钠等。
药学可接受的盐的相应的平衡离子可以使用各种方法分析和鉴定，所述方法包括但不限于离子交换色谱、离子色谱、毛细管电泳、电感耦合等离子体、原子吸收光谱、质谱或它们的任何组合。
使用以下技术的至少一种回收所述盐：过滤、用非溶剂沉淀接着过滤、溶剂蒸发，或水溶液的情况下使用冻干法。
筛选和表征药学可接受的盐、多晶型和/或溶剂化物可以使用多种技术完成，所述技术包括但不限于热分析、X射线衍射、光谱、显微镜方法、元素分析。使用的各种光谱技术包括但不限于Raman、FTIR、UVIS和NMR(液体和固体状态)。各种显微镜技术包括但不限于IR显微镜检术和拉曼(Raman)显微镜检术。
本发明的药物组合物
本申请还提供药物组合物，其包含至少一种式(I)的化合物或所述化合物的药学可接受的盐、溶剂化物、酯、酸、药物活性代谢物或前药、以及药学可接受的载体或赋形剂、以及任选的其它治疗剂。
在治疗过程中，可以根据情况单独或与一种或多种其它的治疗剂组合使用。可以通过注射、口服、吸入、直肠和经皮施用中的至少一种将包含本发明化合物的药物施用给患者。其它的治疗剂可以选自以下药物：免疫抑制剂(例如他克莫司、环抱菌素、雷帕霉素、甲氨蝶岭、环磷酰胺、硫唑嘌呤、巯嘌呤、麦考酚酯或FTY720)、糖皮质激素类药(例如***、醋酸可的松、***龙、甲泼尼龙、***、倍他米松、曲安西龙、氢羟强的松龙、倍氯米松、醋酸氟氢可的松、醋酸脱氧皮质酮、醛固酮)、非甾体抗炎药(例如水杨酸盐、芳基烷酸、2-芳基丙酸、N-芳基邻氨基苯甲酸、昔康类、考昔类或硫酰替苯胺)、***反应疫苗、抗组胺药、抗白三烯药、β-激动剂、茶碱、抗胆碱药或其它选择性激酶抑制剂(例如mTOR抑制剂、c-Met抑制剂)或her2抗体-药物。另外，所提及的其它治疗剂还可以是雷帕霉素(Rapamycin)、克唑替尼(Crizotinib)、他莫昔芬、雷洛昔芬、阿那曲唑、依西美坦、来曲唑、赫赛汀 TM(曲妥珠单抗)、格列卫 TM(伊马替 尼)、紫杉醇 TM(紫杉醇)、环磷酰胺、洛伐他汀、美诺四环素(Minosine)、阿糖胞苷、5-氟尿嘧啶(5-FU)、甲氨蝶呤(MTX)、紫杉特尔 TM(多西他赛)、诺雷德 TM(戈舍瑞林)、长春新碱、长春碱、诺考达唑、替尼泊苷、依托泊苷、健择 TM(吉西他滨)、埃博霉素(Epothilone)、诺唯本、喜树碱、柔红霉素(Daunonibicin)、更生霉素、米托蒽醌、安吖啶、多柔比星(亚德里亚霉素)、表柔比星或伊达比星。或者，其它治疗剂也可以是细胞因子例如G-CSF(粒细胞集落刺激因子)。或者，其它治疗剂也可以是，例如但不限于，CMF(环磷酰胺、甲氨蝶呤和5-氟尿嘧啶)、CAF(环磷酰胺、亚德里亚霉素和5-氟尿嘧啶)、AC(亚德里亚霉素和环磷酰胺)、FEC(5-氟尿嘧啶、表柔比星和环磷酰胺)、ACT或ATC(亚德里亚霉素、环磷酰胺和紫杉醇)或CMFP(环磷酰胺、甲氨蝶呤、5-氟尿嘧啶和***)。
在本发明的实施方式中，在根据本发明对患者进行治疗时，给定药物的量取决于诸多因素，如具体的给药方案、疾病或病症类型及其严重性、需要治疗的受治疗者或宿主的独特性(例如体重)，但是，根据特定的周围情况，包括例如已采用的具体药物、给药途径、治疗的病症、以及治疗的受治疗者或宿主，施用剂量可由本领域已知的方法常规决定。通常，就成人治疗使用的剂量而言，施用剂量典型地在0.02-5000mg/天，例如约1-1500mg/天的范围。该所需剂量可以方便地被表现为一剂、或同时给药的(或在短时间内)或在适当的间隔的分剂量，例如每天二、三、四剂或更多分剂。本领域技术人员可以理解的是，尽管给出了上述剂量范围，但具体的有效量可根据患者的情况并结合医师诊断而适当调节。
本发明的药物的用途
本发明的激酶抑制剂包括式(I)的化合物或其药学可接受的盐、溶剂化物、酯、酸、代谢物或前药、或药物组合物，用于降低或抑制细胞或受试者的cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2激酶活性，并且/或者在受试者中预防或治疗cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2活性 相关病症。
式(I)的化合物或其药学可接受的盐、溶剂化物、酯、酸、代谢物或前药，或其药物组合物可用于治疗、预防或改善一种或多种选自下组的疾病：实体瘤(包括良性或者尤其恶性类型)、尤其肉瘤、胃肠道间质肿瘤(Gastrointestinal Stromal Tumors，GIST)、结直肠癌(colon cancer)、急性粒细胞白血病(Acute Myeloblastic Leukemia，AML)、慢性髓性白血病(Chronic Myelogenous Leukemia，CML)、瘤形成、甲状腺癌、***性肥大细胞病、嗜酸性粒细胞增多综合征、纤维变性、红斑狼疮、移植物抗宿主病、神经纤维瘤、肺高压、阿尔茨海默病、***瘤、无性细胞瘤、肥大细胞肿瘤、肺癌、支气管癌、睾丸上皮内瘤形成、黑色素瘤、乳癌、神经母细胞瘤、***状/滤泡型甲状腺癌、恶性淋巴瘤、非霍奇金淋巴瘤、2型多发性内分泌瘤形成、嗜铬细胞瘤、甲状腺癌、甲状旁腺增生/腺瘤、结肠癌、结肠直肠腺瘤、卵巢癌、***癌、成胶质细胞瘤、脑肿瘤、恶性神经胶质瘤、胰腺癌、恶性胸膜间皮瘤、成血管细胞瘤、血管瘤、肾癌、肝癌、肾上腺癌、膀胱癌、胃癌、直肠癌、***癌、***、子宫内膜癌、多发性骨髓瘤、颈和头部肿瘤、以及其他增生性或增殖性疾病或类似疾病、或其组合。特别优选治疗胃肠道间质瘤、结直肠癌、急性粒细胞白血病、慢性髓性白血病、甲状腺癌或类似疾病、或其组合。最优选地，本发明的抑制剂或其药物组合物可用于治疗或预防胃肠道间质瘤，特别是cKIT-T670I突变型的胃肠道间质瘤。
式(I)所述化合物或其药学可接受的盐、溶剂化物、酯、酸、代谢物或前药，或其药物组合物可用于治疗、预防或改善选自下组的自身免疫性疾病：关节炎、风湿性关节炎、骨关节炎、狼疮、类风湿性关节炎、炎性肠病、银屑病性关节炎、骨关节炎、斯蒂尔病(Still's disease)、青少年关节炎、糖尿病、重症肌无力症、桥本甲状腺炎(Hashimoto's thyroiditis)、奥德甲状腺炎(Ord's hyroiditis)、格雷夫斯病(Graves'disease)、类风湿性关节炎综合征(
Figure PCTCN2018115025-appb-000006
syndrome)、多发性硬化症、传染性神经元炎(Guillain-Barrésyndrome)、急性播散性脑脊髓炎、阿狄森病(Addison's disease)、视性眼阵孪-肌阵孪综合征、强直性脊椎炎、抗磷脂抗体综合征、再生障碍性贫血、自身免疫 性肝炎、乳糜泻(coeliac disease)、古德帕斯彻综合征(Goodpasture's syndrome)、特发性血小板减少性紫癜、视神经炎、硬皮病、原发性胆汁性肝硬化、莱特尔综合征(Reiter's syndrome)、高安动脉炎(Takayasu's arteritis)、颞动脉炎、温型自身免疫性溶血性贫血、韦格纳肉芽肿病(Wegener's granulomatosis)、银屑病、全身脱毛、贝赫切特病(Behcet's disease)、慢性疲劳、家族性自主神经功能异常、子宫内膜异位、间质性膀胱炎、神经肌强直、硬皮病或外阴痛。
化合物的制备
使用本领域技术人员已知的标准合成技术或使用本领域已知的方法与本文描述的方法组合，可以合成式(I)的化合物。另外，本文给出的溶剂、温度和其它反应条件可以根据本领域技术而改变。作为进一步的指导，也可以利用以下的合成方法。
所述反应可以按顺序使用，以提供本文描述的化合物；或它们可以用于合成片段，所述片段通过本文描述的方法和/或本领域已知的方法随后加入。
在某些实施方式中，本文提供的是本文描述的激酶抑制剂化合物的制备方法及其使用方法。在某些实施方式中，本文描述的化合物可以使用以下合成的方案合成。可以使用与下述类似的方法，通过使用适当的可选择的起始原料，合成化合物。
用于合成本文描述的化合物的起始原料可以被合成或可以从商业来源获得。本文描述的化合物和其它相关具有不同取代基的化合物可以使用本领域技术人员已知的技术和原料合成。制备本文公开的化合物的一般方法可以来自本领域已知的反应，并且该反应可以通过由本领域技术人员所认为适当的试剂和条件修改，以引入本文提供的分子中的各种部分。
如果需要，反应产物可以使用常规技术分离和纯化，包括但不限于过滤、蒸馏、结晶、色谱等方法。这些产物可以使用常规方法表征，包括物理常数和图谱数据。
制备式(I)的化合物的合成方案的非限制性实施例参见以下合成路线。
实施例1：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻唑 -2-基)-4-((4-甲基哌嗪-1-基)甲基)苯甲酰胺
Figure PCTCN2018115025-appb-000007
N-(4-甲基噻唑-2-基)乙酰胺：在100mL圆底烧瓶中加入4-甲基噻唑-2-胺(2g)后加入无水二氯甲烷(50mL)、三乙胺(3.9mL)，慢慢滴加乙酰氯(1.5mL)。反应体系在室温下、氩气保护反应4小时。反应结束后，体系在减压下蒸干溶剂，所得物用饱和碳酸氢钠中和至pH>10后，用乙酸乙酯萃取。有机相分别用水、饱和食盐水洗涤后用无水硫酸钠干燥。有机相经过滤，减压蒸干后得粗品。粗品经加压硅胶柱层析提纯后得纯品，MS(ESI)m/z(M+1)+：157.05。
(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1-(四氢-2H-吡喃-2-基)-1H-吲唑-6-基)噻唑-2-基)乙酰胺：在圆底烧瓶中加入N-(4-甲基噻唑-2-基)乙酰胺(1.0g)后加入二甲亚砜(20mL)、(E)-6-碘-3-(2-(吡啶-2-基)乙烯基)-1-(四氢-2H-吡喃-2-基)-1H-吲唑(3.1g)、四三苯基膦钯(0.37g)和碳酸铯(6.2g)。反应体系在氩气保护下加热至130℃反应14小时。反应结束后，体系在减压下蒸干溶剂，所得物用水稀释后用乙酸乙酯萃取。有机相分别用水、饱和食盐水洗涤后用无水硫酸钠干燥。有机相经过滤，减压蒸干后得粗品。粗品经加压硅胶柱层析提纯后得纯品，MS(ESI)m/z(M+1)+：460.18。
(E)-4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻唑-2-胺：在100mL圆底烧瓶中加入(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1-(四氢-2H-吡喃-2-基)-1H-吲唑-6-基)噻唑-2-基)乙酰胺(2.0g)、乙醇(20mL)和6摩尔/升盐酸(15mL)。反应体系在氩气保护下加热至90℃反应14 小时。反应结束后，体系在减压下蒸干溶剂，所得物用饱和碳酸氢钠中和至pH>10后，有固体析出，过滤，得粗品。粗品经乙酸乙酯洗涤，得纯品，MS(ESI)m/z(M+1)+：334.11。
(E)-6-(2-氨基-4-甲基噻唑-5-基)-3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-1-甲酸叔丁酯：在50mL圆底烧瓶中加入(E)-4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻唑-2-胺(1g)后加入无水N,N-二甲基甲酰胺(10mL)、三乙胺(0.9mL)、二碳酸二叔丁酯(1.0g)。反应体系在室温下、氩气保护反应4小时。反应结束后，体系在减压下蒸干溶剂，所得物用饱和碳酸氢钠中和至pH>10后用乙酸乙酯萃取水相。有机相分别用水、饱和食盐水洗涤后用无水硫酸钠干燥。有机相经过滤，减压蒸干后得粗品。粗品经加压硅胶柱层析提纯后得纯品，MS(ESI)m/z(M+1)+：434.17。
(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻唑-2-基)-4-((4-甲基哌嗪-1-基)甲基)苯甲酰胺(1)：在圆底烧瓶中加入(E)-6-(2-氨基-4-甲基噻唑-5-基)-3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-1-甲酸叔丁酯(0.05g)后加入N,N-二甲基甲酰胺(5mL)、4-((4-甲基哌嗪-1-基)甲基)苯甲酸(0.03g)、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(0.07g)和三乙胺(0.03mL)。反应体系在氩气保护下室温搅拌14小时。反应结束后，体系在减压下蒸干溶剂，所得物用水稀释后用乙酸乙酯萃取。有机相分别用水、饱和食盐水洗涤后用无水硫酸钠干燥。有机相经过滤，减压蒸干后得粗品。
粗品溶于无水二氯甲烷(2毫升)，加入三氟乙酸(1毫升)。反应体系在氩气保护下室温搅拌14小时。反应结束后，体系在减压下蒸干溶剂，所得物用水稀释后用饱和碳酸氢钠溶液中和至pH>10。水相用乙酸乙酯萃取，有机相分别用水、饱和食盐水洗涤后用无水硫酸钠干燥。有机相经过滤，减压蒸干后得粗品。粗品经加压硅胶柱层析提纯后得化合物1，MS(ESI)m/z(M+1)+：550.24。
实施例2：(E)-4-氯-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基) 噻唑-2-基)-3-(三氟甲基)苯甲酰胺
Figure PCTCN2018115025-appb-000008
实施例2的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：540.09。
实施例3：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻唑 -2-基)-2-(3-(三氟甲基)苯基)乙酰胺
Figure PCTCN2018115025-appb-000009
实施例3的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：520.14。
实施例4：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻唑 -2-基)异烟酰胺
Figure PCTCN2018115025-appb-000010
实施例4的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：439.14。
实施例5：(E)-N-(5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻唑-2-基) 乙酰胺
Figure PCTCN2018115025-appb-000011
实施例5的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：361.11。
实施例6：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻唑 -2-基)-3-(三氟甲基)苯甲酰胺
Figure PCTCN2018115025-appb-000012
实施例6的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：506.13。
实施例7：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻唑 -2-基)-2-(萘-1-基)乙酰胺
Figure PCTCN2018115025-appb-000013
实施例7的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：502.17。
实施例8：(E)-2-(3,4-甲氧基苯基)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯 基)-1H-吲唑-6-基)噻唑-2-基)乙酰胺
Figure PCTCN2018115025-appb-000014
实施例8的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：512.18。
实施例9：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻唑 -2-基)-2-(4-甲基哌嗪-1-基)乙酰胺
Figure PCTCN2018115025-appb-000015
实施例9的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：474.21。
实施例10：(E)-2-(3-氯苯基)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H- 吲唑-6-基)噻唑-2-基)乙酰胺
Figure PCTCN2018115025-appb-000016
实施例10的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：486.12。
实施例11：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻 唑-2-基)-2-(哌啶-4-基)乙酰胺
Figure PCTCN2018115025-appb-000017
实施例11的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：459.20。
实施例12：(E)-2-(二甲氨基)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H- 吲唑-6-基)噻唑-2-基)乙酰胺
Figure PCTCN2018115025-appb-000018
实施例12的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：419.17。
实施例13：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻 唑-2-基)-2-(哌啶-3-基)乙酰胺
Figure PCTCN2018115025-appb-000019
实施例13的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：459.20。
实施例14:(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻 唑-2-基)哌啶-4-甲酰胺
Figure PCTCN2018115025-appb-000020
实施例14的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：445.18。
实施例15：(E)-2-氨基-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6- 基)噻唑-2-基)-4-(甲硫基)丁酰胺
Figure PCTCN2018115025-appb-000021
实施例15的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：465.16。
实施例16：(E)-1-甲基-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6- 基)噻唑-2-基)哌啶-4甲酰胺
Figure PCTCN2018115025-appb-000022
实施例16的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：458.20。
实施例17：(E)-2-氨基-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6- 基)噻唑-2-基)丙酰胺
Figure PCTCN2018115025-appb-000023
实施例17的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：405.15。
实施例18：(E)-2-氨基-4-甲基-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H- 吲唑-6-基)噻唑-2-基)戊酰胺
Figure PCTCN2018115025-appb-000024
实施例18的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：447.20。
实施例19：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻 唑-2-基)异丁酰胺
Figure PCTCN2018115025-appb-000025
实施例19的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：404.16。
实施例20：(E)-2-氨基-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6- 基)噻唑-2-基)-3-苯基丙酰胺
Figure PCTCN2018115025-appb-000026
实施例20的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：481.18。
实施例21：N-(4-甲基-5-(3-(1-甲基-1H-吡唑-4-基)-1H-吲唑-6-基)噻唑-2- 基)-2-(4-甲基哌嗪-1-基)乙酰胺
Figure PCTCN2018115025-appb-000027
2-(4-甲基哌嗪-1-基)-N-(4-甲基噻唑-2-基)乙酰胺(21a)：在圆底烧瓶中加入4-甲基噻唑-2-胺(1.0g)后加入N,N-二甲基甲酰胺(20mL)、2-(4-甲基哌嗪-1-基)乙酸(1.5g)、2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(5.0g)和三乙胺(2.5mL)。反应体系在氩气保护下室温搅拌14小时。反应结束后，体系在减压下蒸干溶剂，所得物用水稀释后用乙酸乙酯萃取。有机相分别用水、饱和食盐水洗涤后用无水硫酸钠干燥。有机相经过滤，减压蒸干后得粗品。粗品经加压硅胶柱层析提纯后得纯品，MS(ESI)m/z(M+1)+：255.13。
N-(4-甲基-5-(1-(四氢-2H-吡喃-2-基)-1H-吲唑-6-基)噻唑-2-基)-2-(4-甲基哌嗪-1-基)乙酰胺(21b)：在圆底烧瓶中加入2-(4-甲基哌嗪-1-基)-N-(4-甲基噻唑-2-基)乙酰胺(1.0g)后加入二甲亚砜(10mL)、6-碘-1-(四氢-2H-吡喃-2-基)-1H-吲唑(1.3g)、四三苯基膦钯(0.45g)和碳酸铯(3.8g)。反应体系在氩气保护下加热至130℃反应14小时。反应结束后，体系在减压下蒸干溶剂，所得物用水稀释后用乙酸乙酯萃取。有机相分别用水、饱和食盐水洗涤后用无水硫酸钠干燥。有机相经过滤，减压蒸干后得粗品。粗品经加压硅胶柱层析提纯后得纯品，MS(ESI)m/z(M+1)+：455.23。
N-(5-(1H-吲唑-6-基)-4-甲基噻唑-2-基)-2-(4-甲基哌嗪-1-基)乙酰胺(21c)：在50mL圆底烧瓶中加入N-(4-甲基-5-(1-(四氢-2H-吡喃-2-基)-1H-吲唑-6-基)噻唑-2-基)-2-(4-甲基哌嗪-1-基)乙酰胺(1.0g)、乙醇(10mL)和6摩尔/升盐酸(5mL)。反应体系在氩气保护下加热至90℃反应14小时。反应结束后，体系在减压下蒸干溶剂，所得物用饱和碳酸氢钠中和至pH>10后，有固体析出，过滤，得粗品。粗品经乙酸乙 酯洗涤，得纯品，MS(ESI)m/z(M+1)+：371.17。
N-(5-(3-碘-1H-吲唑-6-yl)-4-甲基噻唑-2-基)-2-(4-甲基哌嗪-1-基)乙酰胺(21d)：在50mL圆底烧瓶中加入N-(5-(1H-吲唑-6-基)-4-甲基噻唑-2-基)-2-(4-甲基哌嗪-1-基)乙酰胺(0.6g)后加入N,N-二甲基甲酰胺(10mL)、碘(0.8g)和氢氧化钾(0.4g)。反应体系在氩气保护下室温搅拌8小时。反应结束后，体系在减压下蒸干溶剂，所得物用水稀释后，水相用乙酸乙酯萃取，有机相分别用水、饱和食盐水洗涤后用无水硫酸钠干燥。有机相经过滤，减压蒸干后得粗品。粗品经加压硅胶柱层析提纯后得纯品，MS(ESI)m/z(M+1)+：497.06。
N-(5-(1-乙酰基-3-碘-1H-吲唑-6-基)-4-甲基噻唑-2-基)-2-(4-甲基哌嗪-1-基)乙酰胺(21e)：在50mL圆底烧瓶中加入N-(5-(3-碘-1H-吲唑-6-yl)-4-甲基噻唑-2-基)-2-(4-甲基哌嗪-1-基)乙酰胺(0.5g)后加入无水二氯甲烷(10mL)、三乙胺(0.3mL)、乙酰氯(0.1g)。反应体系在室温下、氩气保护反应4小时。反应结束后，体系在减压下蒸干溶剂，所得物用饱和碳酸氢钠中和至pH>10后用乙酸乙酯萃取。有机相分别用水、饱和食盐水洗涤后用无水硫酸钠干燥。有机相经过滤，减压蒸干后得粗品。粗品经加压硅胶柱层析提纯后得纯品，MS(ESI)m/z(M+1)+：538.07。
N-(4-甲基-5-(3-(1-甲基-1H-吡唑-4-基)-1H-吲唑-6-基)噻唑-2-基)-2-(4-甲基哌嗪-1-基)乙酰胺(21)：在圆底烧瓶中加入N-(5-(1-乙酰基-3-碘-1H-吲唑-6-基)-4-甲基噻唑-2-基)-2-(4-甲基哌嗪-1-基)乙酰胺(0.1g)后加入1,4-二氧六环(10mL)、水(2mL)、(1-甲基-1H-吡唑-4-基)硼酸(0.03g)、四三苯基膦钯(0.02g)和碳酸钾(0.07g)。反应体系在氩气保护下加热至80℃反应14小时。反应结束后，体系在减压下蒸干溶剂，所得物用水稀释后用乙酸乙酯萃取。有机相分别用水、饱和食盐水洗涤后用无水硫酸钠干燥。有机相经过滤，减压蒸干后得粗品。粗品经加压硅胶柱层析提纯后得化合物21，MS(ESI)m/z(M+1)+：451.21。
实施例22：(E)-2-羟基-4-甲基-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H- 吲唑-6-基)噻唑-2-基)戊酰胺
Figure PCTCN2018115025-appb-000028
实施例22的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：448.18。
实施例23：N-(5-(3-(2-氨基嘧啶-5-基)-1H-吲唑-6-基)-4-甲基噻唑-2- 基)-2-(4-甲基哌嗪-1-基)乙酰胺
Figure PCTCN2018115025-appb-000029
实施例23的合成通过使用类似于实施例21中所述的步骤完成。MS(ESI)m/z(M+1)+：464.20。
实施例24：N-(4-甲基-5-(3-(吡啶-3-基)-1H-吲唑-6-基)噻唑-2-基)-2-(4- 甲基哌嗪-1-基)乙酰胺
Figure PCTCN2018115025-appb-000030
实施例24的合成通过使用类似于实施例21中所述的步骤完成。MS(ESI)m/z(M+1)+：448.19。
实施例25：N-(5-(3-(5-氟吡啶-3-基)-1H-吲唑-6-基)-4-甲基噻唑 -2-yl)-2-(4-甲基哌嗪-1-基)乙酰胺
Figure PCTCN2018115025-appb-000031
实施例25的合成通过使用类似于实施例21中所述的步骤完成。MS(ESI)m/z(M+1)+：466.18。
实施例26：N-(5-(3-(3-氟苯基)-1H-吲唑-6-基)-4-甲基噻唑-2-基)-2-(4-甲 基哌嗪-1-基)乙酰胺
Figure PCTCN2018115025-appb-000032
实施例26的合成通过使用类似于实施例21中所述的步骤完成。MS(ESI)m/z(M+1)+：465.19。
实施例27：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻 唑-2-基)-2-(1-甲基哌啶-4-基)乙酰胺
Figure PCTCN2018115025-appb-000033
实施例27的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：473.21。
实施例28：N-(4-甲基-5-(3-(吡啶-3-基)-1H-吲唑-6-基)噻唑-2- 基)-2-(3-(三氟甲基)苯基)乙酰胺
Figure PCTCN2018115025-appb-000034
实施例28的合成通过使用类似于实施例21中所述的步骤完成。MS(ESI)m/z(M+1)+：494.13。
实施例29：(E)-1-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻 唑-2-基)-3-(3-(三氟甲基)苯基)脲
Figure PCTCN2018115025-appb-000035
实施例29的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：521.14。
实施例30：(E)-N-(4-甲基-5-(3-(2-(吡啶-2-基)乙烯基)-1H-吲唑-6-基)噻 唑-2-基)乙酰胺
Figure PCTCN2018115025-appb-000036
实施例30的合成通过使用类似于实施例1中所述的步骤完成。MS(ESI)m/z(M+1)+：376.13。
实施例31：对癌细胞增殖的影响
通过测试本发明的化合物对癌细胞生长的影响(表2)，进一步评估文中化合物对癌细胞增殖的抑制作用、及其对抑制癌细胞增殖的选择性。
本实施例中选用了人胃肠道间质瘤细胞系细胞GIST-T1(表达野生型C-KIT基因)(购自Cosmo Bio Co.,Ltd.(日本))、人胃肠道间质瘤细胞系细胞GIST-T1-T670I(表达C-KIT-T670I突变基因)(由本实验室用CRISPR技术构建)、小鼠原B细胞BaF3(购自ATCC)。此外，本实施例还选用了小鼠Tel-cKit-BaF3(稳定表达C-KIT野生型激酶)、小鼠Tel-cKit/T670I-BaF3(稳定表达cKIT T670I突变型激酶)、小鼠Tel-PDGFRα-BaF3(稳定表达PDGFRα激酶)、小鼠Tel-PDGFRβ-BaF3(稳定表达PDGFRβ激酶)、小鼠Tel-VEGFR2-BaF3(稳定表达VEGFR2激酶)、小鼠Tel-FLT3-BaF3(稳定表达FLT3激酶)。上述细胞系均由本实验室构建，构建方法为：经PCR分别扩增人类C-KIT、C-KIT T670I、PDGFRα、PDGFRβ、VEGFR2、FLT3激酶区序列，并分别***到带有N端TEL片段和/或NPM片段和/或TPR片段的MSCV-Puro载体(购自Clontech)，通过逆转录病毒方法，稳定转入小鼠BaF3细胞，并且撤除IL-3生长因子，最终得到依赖C-KIT、C-KIT/T670I、PDGFRα、PDGFRβ、VEGFR2、FLT3转入蛋白的细胞系。GIST-T1-T670I(表达C-KIT-T670I突变基因)细胞系由本实验室构建，构建方法为：靶向KIT基因T670位点附近的sgRNA由美国麻省理工学院张锋实验室的CRISPR设计工具设计(网址：crispr.mit.edu)，并克隆进pSpCas9(BB)-2A-Puro载体(Addgene，美国)；所获得的载体与T670位点附近带有T670I位点突变的一段单链寡聚核苷酸共同转染细胞，经过抗生素筛选后，稀释并在96孔板中进行单细胞培养；利用Sanger测序法对细胞T670位点进行测序检测验证。
在实施例中将不同浓度(0.000508μM、0.00152μM、0.00457μM、0.0137μM、0.0411μM、0.123μM、0.370μM、1.11μM、3.33μM、10μM于DMSO中)的本发明化合物及对照化合物阿西替尼Axitinib(购自中国，MedChem Express)分别加入到上述细胞中，并孵育72小时，通过CCK-8(购自贝博生物公司，中国上海)细胞活力检测试剂盒(CCK-8可被活细胞中的脱氢酶还原为具有高度水溶性的黄色甲瓒产物，生成的甲瓒物数量与活细胞的数量成正比)对孵育后的细胞进行检测，通过酶标仪对活细胞的数目进行定量，并计算各个化合物和对照化合物的GI 50(结果示于表2和表3中)。
表2所示的实验结果表明，本发明的化合物对突变型cKIT-T670I、VEGFR2、PDGFRα、PDGFRβ、FLT3都有一定的抑制作用，尤其是与cKIT野生型相比对突变型cKIT-T670I具有更强的抑制作用。与Axitinib相比，本发明的优选化合物针对突变型cKIT-T670I具有相当的或更强的抑制活性，但是对野生型cKIT的抑制活性相对较弱。cKIT野生型在正常的造血干细胞的发育早期起着非常重要的作用，因此在不必要的情况下，抑制cKIT激酶会产生机理性的毒性，并且有文献报道同时抑制FLT3和cKIT会引起骨髓抑制毒性。另外，Axitinib对母本BaF3细胞有一定抑制作用，并且在野生型cKIT和突变型cKIT-T670I之间也没有选择性；而本发明的化合物在突变型cKIT-T670I与野生型cKIT和母本BaF3细胞之间表现出明显的选择性抑制，这表明本发明的优选化合物在抑制突变型cKIT-T670I的同时不会由于抑制野生型cKIT和FLT3而产生骨髓抑制毒性的问题。
表2
Figure PCTCN2018115025-appb-000037
Figure PCTCN2018115025-appb-000038
如表3所示，以本发明的化合物7和9与对照化合物伊马替尼Imatinib(购自中国，MedChem Express)对胃肠间质瘤细胞系GIST-T1以及本实验室构建的Imaitinib耐药突变的GIST-T1-T670I细胞系上进行测试发现，本发明的化合物不仅对Imatinib敏感的胃肠间质瘤有很强的抑制作用，对Imatinib耐药的GIST-T1-T670I也有很强的抑制作用。这表明本发明的化合物能够用于治疗T670I突变的胃肠间质瘤。
表3
GI 50(μM) GIST-T1 GIST-T1-T670I
Imaitnib 0.010 >10
7 0.032 0.065
9 0.019 0.039
实施例32：动物实验
在本实施例中，分别测试化合物9在TEL-cKIT/T670I-BaF3及GIST-T1-T670I的小鼠模型中的实验结果。
实验步骤如下：
(1)从北京维通利华实验动物有限责任公司购买饲养4-6周龄的Bal b/c雌性小鼠，饲养于SPF级实验室中，饮水及垫料均经高压消毒无菌处理，有关小鼠的所有操作均在无菌条件下进行。
(2)第0天分别在所有小鼠左侧背部皮下分别注入约5×10 6个TEL-cKIT/T670I-BaF3或5×10 6个GIST-T1-T670I细胞。
(3)对于TEL-cKIT/T670I-BaF3的小鼠模型从第6天开始，每天使对应小鼠口服给药甲基纤维素(HKI)溶媒(5只小鼠)；剂量为10 mg/kg、20mg/kg、40mg/kg、100mg/kg鼠重的化合物9(各5只小鼠)；剂量为40mg/kg鼠重的舒尼替尼Sunitinib(购自MedChemExpress，中国)(5只小鼠)。对于GIST-T1-T670I的小鼠模型从第15天开始，每天使对应小鼠口服给药甲基纤维素(HKI)溶媒(5只小鼠)；剂量为剂量为20mg/kg、30mg/kg、40mg/kg的化合物9(各5只小鼠)；剂量为40mg/kg鼠重的舒尼替尼(5只小鼠)。
(4)分别从第6天(TEL-cKIT/T670I-BaF3的小鼠模型)和15天(GIST-T1-T670I的小鼠模型)开始，每天用游标卡尺测量皮下肿瘤的长/宽，并每天记录小鼠体重，分别确定化合物9对小鼠体重的影响。
(5)统计内皮下肿瘤生长趋势，肿瘤体积计算方法：长×宽×宽/2mm 3。
实验结果如图1a-b和2a-b所示。化合物9在TEL-cKIT/T670I-BaF3以及GIST-T1-T670I的小鼠肿瘤模型中，用药剂量为40mg/kg时已经表现出一定的抑制小鼠肿瘤的效果，且随着用药天数的增加，化合物9对小鼠肿瘤的抑制作用愈发显著，抑瘤率高达80％以上，当采用100mg/kg化合物9时在TEL-cKIT/T670I-BaF3小鼠模型中给药后第11天的抑瘤率达100％，当采用40mg/kg化合物9时在GIST-T1-T670I小鼠模型中给药后第28天的抑瘤率为84.3％。化合物9不仅有效地抑制小鼠肿瘤的生长，并且对小鼠的体重基本没有影响，表明化合物9可适用于动物给药。这也证明了本发明的CKIT/T670I抑制剂化合物能够用于治疗T670I突变的胃肠道间质瘤。
本发明提供一种新型激酶抑制剂化合物，其可以用于降低或抑制细胞或受试者的cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2激酶活性，并且/或者在受试者中预防或治疗cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2活性相关病症。因而，可将其制成相应的药物，适于工业应用。
工业应用性
本发明提供一种新型激酶抑制剂化合物，其可以用于降低或抑制细胞或受试者的cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变 型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2激酶活性，并且/或者在受试者中预防或治疗cKIT(特别是突变型cKIT/T670I)、FLT3(包括突变型FLT3-ITD)、PDGFRα、PDGFRβ、和/或VEGFR2活性相关病症。因而，可将其制成相应的药物，适于工业应用。
尽管本文对本发明作了详细说明，但本发明不限于此，本技术领域的技术人员可以根据本发明的原理进行修改，因此，凡按照本发明的原理进行的各种修改都应当理解为落入本发明的保护范围。

Claims (19)

  1. 一种激酶抑制剂，其包括式(I)的化合物或其药学可接受的盐、溶剂化物、酯、酸、代谢物或前体药物，
    Figure PCTCN2018115025-appb-100001
    其中，
    X为-(CH＝CH) m-，其中m为0或1；
    Y选自-NH-或-(CH 2) n-，其中n为0-3的整数；
    R 1选自任选地被1-3个独立的R 4基团取代的苯基、任选地被1-3个独立的R 4基团取代的吡啶基、任选地被1-3个独立的R 4基团取代的吡唑基、和任选地被1-3个独立的R 4基团取代的嘧啶基；
    R 2选自氢和C 1-6烷基；
    R 3选自任选地被1-2个独立的R 5基团取代的C 1-6烷基、C 1-6烷基氨基、以及任选地被1-3个独立的R 4基团取代的苯基、任选地被1-3个独立的R 4基团取代的萘基、任选地被1-3个独立的R 4基团取代的吡啶基、任选地被1-3个独立的R 4基团取代的哌嗪基、和任选地被1-3个独立的R 4基团取代的哌啶基；
    R 4独立地选自卤素、氨基、C 1-6烷基、C 3-6环烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6羟基烷基、C 1-6烷基氨基、C 2-6烷酰胺基、(4-甲基哌嗪-1-基)甲基、吗啉甲基、吗啉基、4-甲基哌嗪-1-基、4-哌啶基、和4-四氢吡喃基；
    R 5独立地选自氨基、羟基、和C 1-6烷硫基。
  2. 根据权利要求1所述的激酶抑制剂，其中X为-(CH＝CH)-。
  3. 根据权利要求1所述的激酶抑制剂，其中Y为直接键或-CH 2-。
  4. 根据权利要求1所述的激酶抑制剂，其中R 1选自任选地被1-3 个独立的R 4基团取代的苯基、吡啶基、吡唑基、和嘧啶基，其中R 4独立地选自卤素、氨基、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基、和(4-甲基哌嗪-1-基)甲基；
  5. 根据权利要求4所述的激酶抑制剂，其中R 1选自任选地被甲基、氨基或卤素取代的苯基、2-吡啶基、3-吡啶基、4-吡唑基、和5-嘧啶基。
  6. 根据权利要求1所述的激酶抑制剂，其中R 2为氢或甲基。
  7. 根据权利要求1所述的激酶抑制剂，其中R 3选自任选地被1-2个独立的R 5基团取代的C 1-6烷基、C 1-6烷基氨基、以及任选地被1-3个独立的R 4基团取代的苯基、萘基、吡啶基、哌嗪基、和哌啶基，其中R 4独立地选自卤素、氨基、C 1-6烷基、C 1-6卤代烷基、C 1-6烷氧基、和(4-甲基哌嗪-1-基)甲基，R 5独立地选自氨基、羟基、和甲硫基。
  8. 根据权利要求7所述的激酶抑制剂，其中R 3选自任选地被氨基、羟基、或甲硫基取代的C 1-6烷基、二甲氨基、任选地被甲基取代的N-哌嗪基、任选地被卤素、三氟甲基、或甲氧基取代的苯基、萘基、4-吡啶基、3-哌啶基、和任选地被甲基取代的4-哌啶基。
  9. 根据权利要求1所述的激酶抑制剂，其中当Y为直接键时，R 3选自任选地被氨基、羟基、或甲硫基取代的C 1-6烷基、和4-吡啶基；当Y为-CH 2-时，R 3选自任选地被甲氧基取代的苯基、任选地被甲基取代的N-哌嗪基、和任选地被甲基取代的4-哌啶基。
  10. 根据权利要求1所述的激酶抑制剂，其中所述化合物选自，
    Figure PCTCN2018115025-appb-100002
    Figure PCTCN2018115025-appb-100003
    Figure PCTCN2018115025-appb-100004
  11. 一种药物组合物，其包括如权利要求1-10中任一项所述的激酶抑制剂，以及药学上可接受的载体或赋形剂，以及任选的其它治疗剂。
  12. 如权利要求1-10中任一项所述的激酶抑制剂在制备用于抑制cKIT、FLT3、PDGFRα、PDGFRβ、和/或VEGFR2激酶活性的药物中的用途。
  13. 如权利要求1-10中任一项所述的激酶抑制剂在制备用于抑制突变型cKIT/T670I激酶活性的药物中的用途。
  14. 如权利要求1-10中任一项所述的激酶抑制剂在制备用于治疗或预防cKIT、FLT3、PDGFRα、PDGFRβ、和/或VEGFR2活性相关疾病、障碍或病症的药物中的用途。
  15. 如权利要求1-10中任一项所述的激酶抑制剂在制备用于治疗或预防突变型cKIT/T670I活性相关疾病、障碍或病症的药物中的用途。
  16. 如权利要求15所述的用途，其中所述疾病、障碍或病症为cKIT-T670I突变型的胃肠道间质瘤。
  17. 如权利要求1-10中任一项所述的激酶抑制剂，用于抑制cKIT、FLT3、PDGFRα、PDGFRβ、和/或VEGFR2激酶活性，特别是用于抑制突变型cKIT/T670I激酶活性。
  18. 如权利要求1-10中任一项所述的激酶抑制剂，用于治疗或预防cKIT、FLT3、PDGFRα、PDGFRβ、和/或VEGFR2活性相关疾病、障碍或病症，特别是用于治疗或预防突变型cKIT/T670I活性相关疾病、障碍或病症。
  19. 如权利要求18所述的PDGFR激酶抑制剂，其中所述疾病、障碍或病症为cKIT-T670I突变型的胃肠道间质瘤。
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