WO2022262699A1 - 取代的苯并咪唑类化合物及包含该化合物的组合物及其用途 - Google Patents

取代的苯并咪唑类化合物及包含该化合物的组合物及其用途 Download PDF

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WO2022262699A1
WO2022262699A1 PCT/CN2022/098557 CN2022098557W WO2022262699A1 WO 2022262699 A1 WO2022262699 A1 WO 2022262699A1 CN 2022098557 W CN2022098557 W CN 2022098557W WO 2022262699 A1 WO2022262699 A1 WO 2022262699A1
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compound
pharmaceutically acceptable
hydrate
prodrug
tautomer
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PCT/CN2022/098557
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English (en)
French (fr)
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王义汉
赵九洋
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深圳市塔吉瑞生物医药有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4184—1,3-Diazoles condensed with carbocyclic rings, e.g. benzimidazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D235/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings
    • C07D235/02—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings condensed with carbocyclic rings or ring systems
    • C07D235/04—Benzimidazoles; Hydrogenated benzimidazoles
    • C07D235/06—Benzimidazoles; Hydrogenated benzimidazoles with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 2
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D235/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings
    • C07D235/02—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings condensed with carbocyclic rings or ring systems
    • C07D235/04—Benzimidazoles; Hydrogenated benzimidazoles
    • C07D235/06—Benzimidazoles; Hydrogenated benzimidazoles with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 2
    • C07D235/10—Radicals substituted by halogen atoms or nitro radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/07—Optical isomers

Definitions

  • the invention belongs to the technical field of medicine, and in particular relates to a substituted benzimidazole compound, a composition containing the compound and uses thereof. More specifically, the present invention relates to certain deuterium-substituted 5-((4-bromo-2-fluorophenyl)amino)-4-fluoro-N-(2-hydroxyethoxy)-1-methyl- Compounds of 1H-benzimidazole-6-carboxamide and derivatives thereof and tautomers, stereoisomers, prodrugs, crystal forms, pharmaceutically acceptable salts, hydrates or solvates.
  • deuterium-substituted compounds and compositions thereof can be used as potent selective inhibitors of MEK1 and MEK2 proteins, and can be used for the treatment of diseases caused by MEK kinases, and these deuterium-substituted compounds have better ADME and pharmacokinetic properties. dynamic properties.
  • the RAS/RAF/MEK/ERK kinase pathway is activated in more than 30% of human cancers.
  • Activation of RAS GTPase (GTPase) proteins stimulates phosphorylation and activation of RAF kinases in response to growth factors, hormones, cytokines, etc. These kinases then phosphorylate and activate the intracellular protein kinases MEK1 and MEK2, which in turn phosphorylate and activate other protein kinases ERK1 and 2.
  • This signaling pathway also known as the mitogen-activated protein kinase (MAPK) pathway or the cytoplasmic cascade, mediates the cellular response to growth signals.
  • the underlying function of this pathway is to link receptor activity at the cell membrane to cytoplasmic or nuclear-targeted modifications that control cell proliferation, differentiation, and survival.
  • the RAF family consists of three related kinases (A-, B- and C-RAF) that act as downstream effectors of RAS.
  • RAS-mediated RAF activation also triggers the activation of MEK1 and MEK2, which subsequently phosphorylate ERK1 and ERK2 (extracellular signal-regulated kinases 1 and 2) at tyrosine-185 and threonine-183.
  • ERK1 and ERK2 extracellular signal-regulated kinases 1 and 2
  • Activated ERK1 and ERK2 change locations and accumulate in the nucleus, where they can phosphorylate various substrates, including transcription factors that control cell growth and survival.
  • the kinase components of signaling cascades are incorporated as potentially important targets for modulating disease progression in cancer and other proliferative diseases.
  • MEK1 and MKE2 are members of a larger family of threonine- and tyrosine-disabled dual-specificity kinases that phosphorylate various MAPKs.
  • MEK1 and MEK2 have unique genetic codes, but they share a high degree of homology (80%) within the C-terminal catalytic kinase domain and most of the N-terminal regulatory region.
  • Oncogenic forms of MEK1 and MEK2 have not been identified in human cancers, but constitutive activation of MEK has been shown to lead to cellular transformation.
  • MEK can also be activated by other oncogenes. So far, the only known substrates for MEK1 and MEK2 are ERK1 and ERK2. In addition to the unique ability to phosphorylate tyrosine and threonine residues, this exceptional substrate specificity places MEK1 and MEK2 at critical points in signaling cascades that would enable the integration of many extracellular signals into the MAPK pathway.
  • RAF may have a prominent role in the development of certain tumors, for example, the activating allele of BRAF has been found in ⁇ 70% of melanoma, 40% of papillary thyroid carcinoma, 30% of low-grade ovarian cancer and 10% of colorectal cancer Identified. Most BRAF mutations are found in the kinase domain, with single substitutions (V600E) accounting for at least 80%. Mutant BRAF proteins activate the RAS/RAF/MEK/ERK kinase pathway either through elevated kinase activity of MEK or through activation of C-RAF.
  • Binimetinib (chemical name is 5-((4-bromo-2-fluorophenyl)amino)-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H-benzimidazole-6- Formamide, which has the following structural formula) is a potent non-ATP competitive, highly selective MEK1/2 inhibitor developed by Array BioPharma, which can inhibit MEK, ERK phosphorylation, and BRAF or KRAS mutations at nanomolar concentrations The growth of cancer cells.
  • ADME absorption, distribution, metabolism and/or excretion
  • the present invention discloses a novel deuterium-substituted benzimidazole compound as an effective MEK1/2 inhibitor, which can inhibit ERK phosphorylation by inhibiting activated MEK, and exhibits a wide range of effects on various cancer models.
  • Anti-tumor activity including melanoma, acute myeloid leukemia, glioma, neurofibroma, non-small cell lung cancer, breast cancer, serous carcinoma, gastrointestinal stromal tumor, non-squamous lung cancer, colorectal cancer , biliary tract cancer, myeloma, etc.
  • the compound of the present invention is combined with other anti-tumor therapeutic agents to treat various cancers.
  • the compounds of the invention also exhibit good solubility and better metabolic stability and/or pharmacokinetic properties.
  • the first aspect of the present invention provides formula (I) compound:
  • Y 1 , Y 2 , Y 3 , Y 4 and Y 5 are each independently selected from hydrogen, deuterium or halogen;
  • R 1 , R 2 , R 3 and R 4 are each independently selected from hydrogen or deuterium;
  • Each X is independently selected from CH3 , CD3 , CHD2 or CH2D ;
  • the additional condition is that the above compounds contain at least one deuterium atom
  • the present invention provides compounds containing the present invention or tautomers, stereoisomers, prodrugs, crystal forms, pharmaceutically acceptable salts, hydrates or solvates and pharmaceutically acceptable Excipients for pharmaceutical compositions.
  • the compound of the invention is provided in said pharmaceutical composition in an effective amount.
  • the compounds of the invention are provided in a therapeutically effective amount.
  • the compounds of the invention are provided in a prophylactically effective amount.
  • the pharmaceutical composition further comprises an additional therapeutic agent.
  • the additional therapeutic agent is selected from one or more of BRAF inhibitors, EGFR inhibitors, EGFR antibodies, immune checkpoint inhibitors or CDK4/6 inhibitors.
  • the BRAF inhibitor is selected from vemurafenib, dabrafenib, encorafenib, (S)-methyl-(1-((4-(3 -(5-Chloro-2-fluoro-3-(methylsulfonylamino)phenyl)-1-(propan-2-yl-d 7 )-1H-pyrazol-4-yl)pyrimidin-2-yl )amino)propan-2-yl)carbamate, (S)-(methyl-d 3 )-(1-((4-(3-(5-chloro-2-fluoro-3-(methyl Sulfonylamino)phenyl)-1-isopropyl-1H-pyrazol-4-yl)pyrimidin-2-yl)amino)propan-2-yl)carbamate, (S)-(methyl- d 3 )-(1-((4-(3-(5-chloro-2-fluoro-3-(
  • the EGFR inhibitor is selected from the group consisting of gefitinib, erlotinib, afatinib, dacomitinib, lapatinib , Osimertinib, Almetinib, Fumetinib, CO-1686, WZ4002, PD153035, PF00299804.
  • the EGFR antibody is selected from cetuximab (cetuximab), panitumumab (panitumumab), necitumumab (Necitumumab).
  • the immune checkpoint inhibitor is selected from Since pembrolizumab, ipilimumab and nivolumab, atezolizumab, avelumab, durvalumab (durvalumab), pidilzumab.
  • the CDK4/6 inhibitor is selected from palbociclib, ribociclib, abemaciclib.
  • the present invention provides a method for preparing the above-mentioned pharmaceutical composition, comprising the following steps: mixing a pharmaceutically acceptable excipient with the compound of the present invention or its tautomer, stereoisomer Body, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate are mixed to form a pharmaceutical composition.
  • the present invention further provides a method of treating MEK kinase-mediated diseases, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention, or a tautomer, stereoisomer, or Construct, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition of the present invention.
  • the MEK kinase-mediated disease is melanoma, acute myeloid leukemia, glioma, neurofibroma, non-small cell lung cancer, breast cancer, serous carcinoma, gastrointestinal stromal tumor , non-squamous lung cancer, colorectal cancer, biliary tract cancer, myeloma.
  • the melanoma is selected from a BRAF V600 mutated melanoma.
  • the colorectal cancer is selected from BRAF V600 mutated colorectal cancer.
  • the BRAF V600 mutation is selected from a BRAF V600E mutation or a BRAF V600K mutation.
  • the neurofibroma is selected from neurofibromatosis type 1 (NF1) or plexiform neurofibroma;
  • deuterated means that one or more hydrogens in a compound or group are replaced by deuterium; deuterated can be monosubstituted, disubstituted, multisubstituted or fully substituted.
  • deuterated can be monosubstituted, disubstituted, multisubstituted or fully substituted.
  • deuterated can be monosubstituted, disubstituted, multisubstituted or fully substituted.
  • one or more deuterated and “one or more deuterated” are used interchangeably.
  • non-deuterated compound refers to a compound containing deuterium atoms in a proportion not higher than the natural deuterium isotope content (0.015%).
  • the term "subject” includes, but is not limited to: human (i.e., male or female of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., Young, middle-aged, or older adults)) and/or non-human animals, e.g., mammals, e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys), cows, pigs, horses , sheep, goats, rodents, cats and/or dogs.
  • the subject is a human.
  • the subject is a non-human animal.
  • treating includes an effect on a subject suffering from a particular disease, disorder or condition, which reduces the severity of the disease, disorder or condition, or delays or slows down the disease, disorder or the development of a condition ("therapeutic treatment"), and also includes effects that occur before a subject begins to suffer from a particular disease, disorder or disease (“prophylactic treatment").
  • an "effective amount" of a compound refers to an amount sufficient to elicit a desired biological response.
  • the effective amount of a compound of the present invention may vary depending on factors such as, for example, the biological target, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age of the subject. Health conditions and symptoms.
  • An effective amount includes therapeutically and prophylactically effective amounts.
  • a "therapeutically effective amount" of a compound is an amount sufficient to provide a therapeutic benefit in the treatment of a disease, disorder, or condition, or to induce one or more effects associated with the disease, disorder, or condition. Symptoms are delayed or minimized.
  • a therapeutically effective amount of a compound refers to that amount of the therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment of a disease, disorder or condition.
  • the term "therapeutically effective amount” can include an amount that improves overall therapy, reduces or avoids symptoms or causes of a disease or disorder, or enhances the therapeutic efficacy of other therapeutic agents.
  • a prophylactically effective amount of a compound is an amount sufficient to prevent a disease, disorder or condition, or an amount sufficient to prevent one or more symptoms associated with a disease, disorder or condition, or to prevent a disease , the number of recurrences of the disorder or condition.
  • a prophylactically effective amount of a compound refers to that amount of the therapeutic agent, alone or in combination with other agents, which provides a prophylactic benefit in the prevention of a disease, disorder or condition.
  • the term “prophylactically effective amount” can include amounts that improve overall prophylaxis, or that enhance the prophylactic efficacy of other prophylactic agents.
  • Combination and related terms refer to the simultaneous or sequential administration of the therapeutic agents of the invention.
  • a compound of the invention can be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms, or simultaneously with another therapeutic agent in a single unit dosage form.
  • the compound of the present invention refers to the following formula (I) and formula (II) compound or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent compounds.
  • the invention relates to compounds of formula (I):
  • Y 1 , Y 2 , Y 3 , Y 4 and Y 5 are each independently selected from hydrogen, deuterium or halogen;
  • R 1 , R 2 , R 3 and R 4 are each independently selected from hydrogen or deuterium;
  • X is selected from CH3 , CD3 , CHD2 or CH2D ;
  • the additional condition is that the above compounds contain at least one deuterium atom
  • the deuterium isotope content of deuterium at the deuterated position is at least 0.015% greater than the natural deuterium isotope content, preferably greater than 30%, more preferably greater than 50%, more preferably greater than 75%, more preferably more than 95%, more preferably more than 99%.
  • the deuterium isotope content of each deuterated position of Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , R 1 , R 2 , R 3 , R 4 and X is at least greater than that of the natural isotope Content 0.015%, more preferably greater than 1%, more preferably greater than 5%, more preferably greater than 10%, more preferably greater than 15%, more preferably greater than 20%, more preferably greater than 25%, more preferably greater than 30%, more preferably greater than 35%, more preferably greater than 40%, more preferably greater than 45%, more preferably greater than 50%, more preferably greater than 55%, more preferably greater than 60%, more preferably greater than 65 %, more preferably greater than 70%, more preferably greater than 75%, more preferably greater than 80%, more preferably greater than 85%, more preferably greater than 90%, more preferably greater than 95%, more preferably greater than 99% .
  • the compound of the present invention contains at least one deuterium atom, more preferably two deuterium atoms, more preferably three deuterium atoms, more preferably four deuterium atoms, more preferably five deuterium atoms deuterium atoms, more preferably six deuterium atoms, more preferably seven deuterium atoms, more preferably eight deuterium atoms, more preferably nine deuterium atoms, more preferably ten deuterium atoms, more preferably Preferably there are eleven deuterium atoms, more preferably twelve deuterium atoms.
  • Y 1 , Y 2 , Y 3 , Y 4 and Y 5 are each independently selected from hydrogen, deuterium or halogen
  • Y 1 is selected from hydrogen, deuterium or halogen
  • Y 2 is selected from hydrogen , deuterium or halogen
  • Y 3 is selected from hydrogen, deuterium or halogen
  • Y 5 is selected from the technical scheme of hydrogen, deuterium or halogen.
  • Y 1 is hydrogen, Y 1 is deuterium or Y 1 is halogen (F, Cl, Br or I)
  • Y 2 is hydrogen, Y 2 is deuterium or Y 2 is halogen (F, Cl, Br or 1 )
  • Y3 is hydrogen, Y3 is deuterium or Y3 is halogen (F, Cl , Br or I)
  • Y5 is hydrogen, Y5 is deuterium or Y5 is halogen (F, Cl , Br or I) the technical scheme.
  • R 1 , R 2 , R 3 and R 4 are each independently selected from hydrogen or deuterium” includes that R 1 is selected from hydrogen or deuterium, R 2 is selected from hydrogen or deuterium, R 3 is selected from Hydrogen or deuterium, and R 4 is selected from the technical scheme of hydrogen or deuterium. More specifically, including R 1 is hydrogen or R 1 is deuterium, R 2 is hydrogen or R 2 is deuterium, R 3 is hydrogen or R 3 is deuterium, and R 4 is hydrogen or R 4 is deuterium.
  • X is selected from CH 3 , CD 3 , CHD 2 or CH 2 D
  • X is CH 3 , CD 3 , CHD 2 or CH 2 D.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , R 1 , R 2 , R 3 and R 4 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein R 1 and R 2 are deuterium and Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , R 3 , R 4 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein R 3 and R 4 are deuterium, Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , R 1 , R 2 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein R 1 , R 2 , R 3 and R 4 are deuterium and Y 1 , Y 2 , Y 3 , Y 4 , Y 5 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein Y 1 is deuterium, Y 2 , Y 3 , Y 4 , Y 5 , R 1 , R 2 , R 3 , R 4 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , Y 1 is deuterium, and Y 2 , Y 3 , Y 4 , Y 5 , R 1 , R 2 , R 3 and R 4 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , R 1 and R 2 are deuterium, and Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , R 3 and R 4 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , R 3 and R 4 are deuterium, and Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , R 1 and R 2 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , R 1 , R 2 , R 3 and R 4 are deuterium, and Y 1 , Y 2 , Y 3 , Y 4 and Y 5 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein Y 1 , R 1 and R 2 are deuterium and Y 2 , Y 3 , Y 4 , Y 5 , R 3 , R 4 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein Y 1 , R 3 and R 4 are deuterium and Y 2 , Y 3 , Y 4 , Y 5 , R 1 , R 2 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein Y 1 , R 1 , R 2 , R 3 and R 4 are deuterium, and Y 2 , Y 3 , Y 4 , Y 5 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , Y 1 , R 1 and R 2 are deuterium, and Y 2 , Y 3 , Y 4 , Y 5 , R 3 and R 4 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , Y 1 , R 3 and R 4 are deuterium, and Y 2 , Y 3 , Y 4 , Y 5 , R 1 and R 2 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , Y 1 , R 1 , R 2 , R 3 and R 4 are deuterium, and Y 2 , Y 3 , Y 4 and Y 5 are as defined above.
  • the present invention relates to compounds of formula (II):
  • Y is selected from hydrogen, deuterium or halogen ;
  • R 1 , R 2 , R 3 and R 4 are each independently selected from hydrogen or deuterium;
  • X is selected from CH3 , CD3 , CHD2 or CH2D ;
  • the additional condition is that the above compounds contain at least one deuterium atom
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , and Y 1 , R 1 , R 2 , R 3 and R 4 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein R 1 and R 2 are deuterium, Y 1 , R 3 , R 4 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein R 3 and R 4 are deuterium, and Y 1 , R 1 , R 2 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein R 1 , R 2 , R 3 and R 4 are deuterium, and Y 1 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein Y 1 is deuterium and R 1 , R 2 , R 3 , R 4 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , Y 1 is deuterium, and R 1 , R 2 , R 3 and R 4 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , R 1 and R 2 are deuterium, and Y 1 , R 3 , R 4 and Y 5 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , R 3 and R 4 are deuterium, and Y 1 , R 1 and R 2 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , R 1 , R 2 , R 3 and R 4 are deuterium, and Y 1 is as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein Y 1 , R 1 and R 2 are deuterium and R 3 , R 4 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein Y 1 , R 3 and R 4 are deuterium and R 1 , R 2 and X are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein Y 1 , R 1 , R 2 , R 3 and R 4 are deuterium and X is as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , Y 1 , R 1 and R 2 are deuterium, and R 3 and R 4 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , Y 1 , R 3 and R 4 are deuterium, and R 1 and R 2 are as defined above.
  • the present invention relates to the above-mentioned compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvent A compound wherein X is CD 3 , Y 1 , R 1 , R 2 , R 3 and R 4 are deuterium.
  • the compound, or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate is selected from any of the following compounds :
  • the compounds of the present invention may include one or more asymmetric centers, and thus may exist in various stereoisomeric forms, eg, enantiomeric and/or diastereomeric forms.
  • the compounds of the invention may be individual enantiomers, diastereoisomers or geometric isomers (eg cis and trans isomers), or may be in the form of a mixture of stereoisomers, Racemic mixtures and mixtures enriched in one or more stereoisomers are included.
  • Isomers can be separated from mixtures by methods known to those skilled in the art, including: chiral high pressure liquid chromatography (HPLC) and formation and crystallization of chiral salts; or preferred isomers can be obtained by prepared by asymmetric synthesis.
  • HPLC high pressure liquid chromatography
  • organic compounds may form complexes with solvents in which they react or from which they are precipitated or crystallized. These complexes are known as "solvates”. When the solvent is water, the complex is called a "hydrate”. The invention covers all solvates of the compounds of the invention.
  • solvate refers to a form of a compound, or a salt thereof, which is associated with a solvent, usually formed by a solvolysis reaction. This physical association may include hydrogen bonding.
  • solvents include water, methanol, ethanol, acetic acid, DMSO, THF, diethyl ether, and the like.
  • Suitable solvates include pharmaceutically acceptable solvates and further include stoichiometric solvates and non-stoichiometric solvates. In some instances, the solvate will be capable of isolation, for example, when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid.
  • “Solvate” includes both solution state solvates and isolatable solvates. Representative solvates include hydrates, ethanolates and methanolates.
  • hydrate refers to a compound that combines with water. Generally, the ratio of the number of water molecules contained in a hydrate of a compound to the number of molecules of the compound in the hydrate is determined.
  • a hydrate of a compound can be represented, for example, by the general formula R.x H 2 O, where R is the compound, and x is a number greater than zero.
  • a given compound may form more than one hydrate type, including, for example, monohydrates (x is 1), lower hydrates (x is a number greater than 0 and less than 1, for example, hemihydrates (R 0.5H2 O)) and polyhydrates (x is a number greater than 1, eg, dihydrate (R ⁇ 2H 2 O) and hexahydrate (R ⁇ 6H 2 O)).
  • the compounds of the invention may be in amorphous or crystalline form (polymorphs). Furthermore, the compounds of the invention may exist in one or more crystalline forms. Accordingly, the present invention includes within its scope all amorphous or crystalline forms of the compounds of the invention.
  • polymorph refers to a crystalline form of a compound (or a salt, hydrate or solvate thereof) in a particular crystal packing arrangement. All polymorphs have the same elemental composition. Different crystalline forms generally have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal shapes, optoelectronic properties, stability and solubility. Recrystallization solvent, crystallization rate, storage temperature, and other factors can cause one crystalline form to predominate. Various polymorphs of a compound can be prepared by crystallization under different conditions.
  • the present invention also includes isotopically labeled compounds which are identical to those of the present invention but wherein one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number normally found in nature.
  • isotopes that may be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F, and 36 Cl.
  • the compounds of the present invention their prodrugs and pharmaceutically acceptable salts of the compounds or the prodrugs containing the above-mentioned isotopes and/or other isotopes of other atoms all belong to the scope of the present invention.
  • Certain isotopically-labeled compounds of the invention eg, those incorporating radioactive isotopes (eg, 3H and14C ), are useful in drug and/or substrate tissue distribution assays. Tritium, ie3H , and carbon- 14 , ie14C isotopes are particularly preferred because of their ease of preparation and detection.
  • isotope-labeled compound of formula (I) of the present invention and its prodrug can generally be prepared in this way.
  • prodrugs are also included within the context of the present invention.
  • the term "prodrug” as used herein refers to a compound that is converted in vivo to its active form having a medical effect, for example by hydrolysis in blood.
  • Pharmaceutically acceptable prodrugs are described in T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, Vol. 14 of A.C.S. Symposium Series, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, and D. Fleisher, S. Ramon, and H. Barbra "Improved oral drug delivery: solubility limitations overcome by the use of prodrugs", Advanced Drug Delivery Reviews (1996) 19(2) 115-130, per intro This article is for reference.
  • a prodrug is any covalently bonded compound of the invention which, when administered to a patient, releases the parent compound in vivo.
  • Prodrugs are generally prepared by modifying functional groups in such a way that the modification can be cleaved by routine manipulation or in vivo to yield the parent compound.
  • Prodrugs include, for example, compounds of the invention wherein a hydroxy, amino, or thiol group is bonded to any group that, when administered to a patient, cleaves to form the hydroxy, amino, or thiol group.
  • representative examples of prodrugs include, but are not limited to, acetate/amide, formate/amide and benzoate/amide derivatives of the hydroxy, sulfhydryl and amino functional groups of the compounds of formula (I).
  • esters such as methyl ester, ethyl ester and the like can be used.
  • the esters themselves may be reactive and/or hydrolyzable under human in vivo conditions.
  • Suitable pharmaceutically acceptable in vivo hydrolyzable ester groups include those which break down readily in the human body to release the parent acid or a salt thereof.
  • Compounds of the invention can be prepared using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes, such as those in the schemes below.
  • the reactions used to prepare the compounds of the present invention can be carried out in suitable solvents, which can be readily selected by those skilled in the art of organic synthesis. Suitable solvents may be substantially nonreactive with the starting materials (reactants), intermediates or products at the temperatures at which the reactions are carried out (eg, temperatures ranging from the solvent's freezing temperature to the solvent's boiling temperature).
  • a given reaction can be carried out in one solvent or a mixture of more than one solvent.
  • a skilled person can select a solvent for a specific reaction step according to the specific reaction step.
  • the preparation of the compounds of the present invention may involve the protection and deprotection of various chemical groups.
  • the need for protection and deprotection and selection of appropriate protecting groups can be readily determined by those skilled in the art.
  • the chemistry of protecting groups can be found in, eg, Wuts and Greene, Protective Groups in Organic Synthesis, 4th ed., John Wiley & Sons: New Jersey, (2006), which is hereby incorporated by reference in its entirety.
  • the compounds of the present invention can be prepared as their individual stereoisomers by reacting a racemic mixture of the compound with an optically active resolving agent to form a pair of diastereoisomeric compounds, separating the diastereomers and recovering the optically pure enantiomers. isomer.
  • Enantiomeric resolution may be performed using diastereomeric derivatives of the compounds of the invention, preferentially dissociable complexes (eg, crystalline diastereomeric salts).
  • Diastereomers have markedly different physical properties (eg, melting points, boiling points, solubilities, reactivities, etc.) and can be readily separated by taking advantage of these dissimilarities.
  • Diastereomers can be separated by chromatography, preferably by separation/resolution techniques based on differences in solubility. The optically pure enantiomer is then recovered, along with the resolving reagents, by any practical means that will not result in racemization.
  • a more detailed description of techniques suitable for the resolution of stereoisomers of compounds starting from racemic mixtures can be found in Jean Jacques, Andre Collet, Samuel H. Wilen, "Enantiomers, Racemates and Resolution” (“Enantiomers, Racemates and Resolutions”), John Wiley And Sons, Inc., 1981.
  • the reaction can be monitored according to any suitable method known in the art.
  • spectroscopic means such as nuclear magnetic resonance (NMR) spectroscopy (e.g. 1 H or 13 C), infrared (IR) spectroscopy, spectrophotometry (e.g. UV-visible), mass spectrometry (MS)) or by chromatography
  • NMR nuclear magnetic resonance
  • IR infrared
  • spectrophotometry e.g. UV-visible
  • MS mass spectrometry
  • Product formation is monitored by methods such as high performance liquid chromatography (HPLC) or thin layer chromatography (TLC).
  • HPLC high performance liquid chromatography
  • TLC thin layer chromatography
  • compositions, formulations and kits are provided.
  • the invention provides pharmaceutical compositions comprising a compound of the invention (also referred to as "active ingredient") and a pharmaceutically acceptable excipient.
  • the pharmaceutical composition comprises an effective amount of a compound of the invention.
  • the pharmaceutical composition comprises a therapeutically effective amount of a compound of the invention.
  • the pharmaceutical composition comprises a prophylactically effective amount of a compound of the invention.
  • a pharmaceutically acceptable excipient used in the present invention refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound formulated together.
  • Pharmaceutically acceptable carriers, adjuvants or vehicles that can be used in the compositions of the present invention include, but are not limited to, ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins (such as human serum albumin Protein), buffer substances (such as phosphate), glycine, sorbic acid, potassium sorbate, partial glyceride mixture of saturated vegetable fatty acids, water, salt or electrolyte (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate , sodium chloride, zinc salts, silica gel, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene- Block polymers, polyethylene glycol
  • kits eg, pharmaceutical packs.
  • kits can include a compound of the invention, another therapeutic agent, and first and second containers (e.g., vials, ampoules, bottles, syringes, and/or dispersible packs or other suitable container).
  • first and second containers e.g., vials, ampoules, bottles, syringes, and/or dispersible packs or other suitable container.
  • provided kits can also optionally include a third container containing a pharmaceutically acceptable excipient for diluting or suspending a compound of the invention and/or other therapeutic agent.
  • a compound of the invention and other therapeutic agent provided in a first container and a second container are combined to form a unit dosage form.
  • formulation examples illustrate representative pharmaceutical compositions that may be prepared in accordance with the present invention.
  • the present invention is not limited to the following pharmaceutical compositions.
  • Exemplary Formulation 1 - Tablet A compound of the invention in dry powder form can be mixed with a dry gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate is added as a lubricant. The mixture is formed in a tablet machine into 0.3-30 mg tablets (each tablet containing 0.1-10 mg of active compound).
  • Exemplary Formulation 2 - Tablet A compound of the invention in dry powder form can be mixed with a dry gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate is added as a lubricant. The mixture is formed into 30-90 mg tablets (each tablet containing 10-30 mg of active compound) in a tablet machine.
  • Exemplary Formulation 3 - Tablet A compound of the invention in dry powder form can be mixed with a dry gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate is added as a lubricant. The mixture is formed in a tablet machine into 90-150 mg tablets (each tablet containing 30-50 mg of active compound).
  • Exemplary Formulation 4 - Tablet A compound of the invention in dry powder form can be mixed with a dry gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate is added as a lubricant. The mixture is formed in a tablet machine into 150-240 mg tablets (each tablet containing 50-80 mg of active compound).
  • Exemplary Formulation 5 - Tablet A compound of the invention in dry powder form can be mixed with a dry gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate is added as a lubricant. The mixture is formed in a tablet machine into 240-270 mg tablets (each tablet containing 80-90 mg of active compound).
  • Exemplary Formulation 6 - Tablet A compound of the invention in dry powder form can be mixed with a dry gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate is added as a lubricant. The mixture is formed in a tablet machine into 270-450 mg tablets (each tablet containing 90-150 mg of active compound).
  • Exemplary Formulation 7 - Tablet A compound of the invention in dry powder form can be mixed with a dry gel binder in a weight ratio of about 1:2. A smaller amount of magnesium stearate is added as a lubricant. The mixture is formed into 450-900 mg tablets (each containing 150-300 mg of active compound) in a tablet machine.
  • Exemplary Formulation 8 - Capsule A compound of the invention in dry powder form can be mixed with a starch diluent in a weight ratio of about 1:1. The mixture is filled into 250 mg capsules (each capsule contains 125 mg of active compound).
  • Exemplary Formulation 9 - Liquid A compound of the invention (125 mg) can be mixed with sucrose (1.75 g) and xanthan gum (4 mg) and the resulting mixture can be blended, passed through a No. 10 mesh U.S. sieve, and then Mix with a previously prepared aqueous solution of microcrystalline cellulose and sodium carboxymethylcellulose (11:89, 50 mg). Sodium benzoate (10 mg), flavor and color were diluted with water and added with stirring. Sufficient water can then be added to give a total volume of 5 mL.
  • Exemplary Formulation 10 - Injection Compounds of the invention can be dissolved or suspended in a buffered sterile saline injectable aqueous medium to a concentration of about 5 mg/mL.
  • parenteral administration as used herein includes subcutaneous administration, intradermal administration, intravenous administration, intramuscular administration, intraarticular administration, intraarterial administration, intrasynovial administration, intrasternal administration , intracerebrospinal administration, intralesional administration, and intracranial injection or infusion techniques.
  • an effective amount of a compound provided herein is administered.
  • the amount of the compound actually administered can be determined by the physician according to the circumstances, including the condition being treated, the route of administration chosen, the compound actually administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, etc. .
  • the compounds provided herein are administered to a subject at risk of developing the condition, typically on the advice and supervision of a physician, at dosage levels as described above.
  • Subjects at risk of developing a particular condition generally include those with a family history of the condition, or those determined by genetic testing or screening to be particularly susceptible to developing the condition.
  • Chronic administration refers to administering a compound or a pharmaceutical composition thereof for a long period of time, for example, 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc., or may continue administration indefinitely, For example, the rest of the subject's life.
  • chronic administration is intended to provide a constant level of the compound in the blood over an extended period of time, eg, within the therapeutic window.
  • compositions may be administered as a bolus injection, eg, in order to increase the concentration of the compound in the blood to effective levels.
  • the bolus dose depends on the target systemic level of the active ingredient through the body, for example, an intramuscular or subcutaneous bolus dose provides slow release of the active ingredient, while a bolus delivered directly into a vein (e.g., by IV intravenous infusion) ) can be delivered more rapidly, so that the concentration of the active ingredient in the blood rises rapidly to effective levels.
  • the pharmaceutical compositions may be administered as a continuous infusion, eg, by IV infusion, to provide a steady state concentration of the active ingredient in the subject's body. Additionally, in other embodiments, a bolus dose of the pharmaceutical composition may be administered first, followed by a continuous infusion.
  • Oral compositions may take the form of bulk liquid solutions or suspensions or bulk powders. More usually, however, the compositions will be presented in unit dosage form for ease of precise dosing.
  • unit dosage form refers to physically discrete units suitable as unitary dosages for human patients and other mammals, each unit containing a predetermined quantity of active material suitable to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient.
  • Typical unit dosage forms include prefilled, premeasured ampoules or syringes for liquid compositions, or pills, tablets, capsules and the like in the case of solid compositions.
  • the compound will generally be a minor component (from about 0.1 to about 50% by weight, or preferably from about 1 to about 40% by weight), with the remainder being various components useful for forming the desired administration form. Carriers or excipients and processing aids.
  • a typical regimen is one to five oral dosages per day, especially two to four oral dosages, typically three oral dosages.
  • each dose provides from about 0.01 to about 20 mg/kg of the compound of the invention, with preferred doses each providing from about 0.1 to about 10 mg/kg, especially about 1 to about 5 mg/kg.
  • the transdermal dose is generally selected in an amount of about 0.01 to about 20% by weight, preferably about 0.1 to about 20% by weight, preferably about 0.1 to about 10% by weight, and more preferably from about 0.5 to about 15% by weight.
  • Injection dosage levels range from about 0.1 mg/kg/hour to at least 10 mg/kg/hour from about 1 to about 120 hours, especially 24 to 96 hours.
  • a preload bolus of about 0.1 mg/kg to about 10 mg/kg or more may also be given in order to achieve adequate steady state levels.
  • the maximum total dose should not exceed approximately 2 g/day.
  • Liquid forms suitable for oral administration may include suitable aqueous or non-aqueous carriers as well as buffering, suspending and dispersing agents, coloring agents, flavoring agents, and the like.
  • the solid form may comprise, for example, any of the following components, or compounds of similar nature: binders, such as microcrystalline cellulose, tragacanth, or gelatin; excipients, such as starch or lactose, disintegrants, For example, alginic acid, Primogel, or corn starch; lubricants, for example, magnesium stearate; glidants, for example, colloidal silicon dioxide; sweeteners, for example, sucrose or saccharin; or flavoring agents, for example, peppermint, water Methyl sylate or orange flavoring.
  • binders such as microcrystalline cellulose, tragacanth, or gelatin
  • excipients such as starch or lactose, disintegrants, For example, alginic acid, Primogel, or corn starch
  • Injectable compositions are typically based on injectable sterile saline or phosphate buffered saline, or other injectable excipients known in the art.
  • the active compound is typically a minor component, often from about 0.05 to 10% by weight, the remainder being injectable excipients and the like.
  • Transdermal compositions are typically formulated as topical ointments or creams containing the active ingredient.
  • the active ingredients When formulated in an ointment, the active ingredients are typically combined with a paraffinic or a water-miscible ointment base.
  • the active ingredients may be formulated in a cream, with, for example, an oil-in-water cream base.
  • Such transdermal formulations are well known in the art, and generally include other ingredients for enhancing the stable skin penetration of the active ingredient or formulation. All such known transdermal formulations and compositions are included within the scope of the present invention.
  • transdermal administration can be achieved using patches of the reservoir or porous membrane type, or various solid matrices.
  • compositions for oral administration, injection or topical administration are representative only. Other materials and processing techniques, etc. are described in Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pennsylvania, Section 8, which is incorporated herein by reference.
  • the compounds of the invention may also be administered in sustained release form, or from a sustained release delivery system.
  • sustained release materials can be found in Remington's Pharmaceutical Sciences.
  • the invention also relates to pharmaceutically acceptable formulations of the compounds of the invention.
  • the formulation comprises water.
  • the formulation comprises a cyclodextrin derivative.
  • the most common cyclodextrins are ⁇ -, ⁇ -, and ⁇ -cyclodextrins composed of 6, 7, and 8 ⁇ -1,4-linked glucose units, respectively, optionally including a or multiple substituents including, but not limited to, methylated, hydroxyalkylated, acylated, and sulfoalkyl ether substitutions.
  • the cyclodextrin is a sulfoalkyl ether ⁇ -cyclodextrin, eg, sulfobutyl ether ⁇ -cyclodextrin, also known as Captisol. See, eg, U.S. 5,376,645.
  • the formulation includes hexapropyl-beta-cyclodextrin (e.g., 10-50% in water).
  • the invention provides a method of treating a disease in a subject, such as a MEK kinase-mediated disease, comprising administering to the subject a compound of the invention or a tautomer, stereoisomer, prodrug thereof , crystal form, pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition of the present invention.
  • a disease in a subject such as a MEK kinase-mediated disease
  • the treatment method can also be combined with other therapies such as radiotherapy, chemotherapy.
  • MEK kinase-mediated diseases include inflammatory diseases, infections, autoimmune disorders, stroke, ischemia, cardiac disorders, neurological disorders, fibrotic disorders, proliferative disorders, hyperproliferative disorders, tumors, Leukemia, neoplasm, cancer, malignancy, metabolic disease and malignancy.
  • the present invention also provides a method for treating MEK kinase-mediated inflammatory diseases, comprising administering the compound of the present invention or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutical acceptable salt, hydrate or solvate, or the pharmaceutical composition of the present invention.
  • MEK kinase-mediated inflammatory diseases include rheumatoid arthritis or multiple sclerosis.
  • the present invention also provides a method for treating MEK kinase-mediated proliferative diseases, comprising administering the compound of the present invention or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutical acceptable salt, hydrate or solvate, or the pharmaceutical composition of the present invention.
  • the MEK kinase-mediated proliferative disease includes cancer, psoriasis, restenosis, autoimmune disease, or atherosclerosis.
  • the present invention also provides a method for treating MEK kinase-mediated cancer, comprising administering the compound of the present invention or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable Accepted salts, hydrates or solvates, or pharmaceutical compositions of the present invention.
  • MEK kinase-mediated cancers include melanoma (e.g., BRAF V600 mutated melanoma), acute myeloid leukemia, glial flow, neurofibromatosis (e.g., neurofibromatosis type 1 (NF1) or plexiform neurofibroma), non-small cell lung cancer, breast cancer, serous carcinoma, gastrointestinal stromal tumor, nonsquamous lung cancer, colorectal cancer (eg, BRAF V600-mutated colorectal cancer), biliary tract Cancer, myeloma.
  • melanoma e.g., BRAF V600 mutated melanoma
  • acute myeloid leukemia glial flow
  • neurofibromatosis e.g., neurofibromatosis type 1 ( NF1) or plexiform neurofibroma
  • non-small cell lung cancer breast cancer, serous carcinoma, gastrointestinal stromal tumor, nonsquamous lung cancer
  • the present invention describes inhibitors of MEK kinases for the treatment of diseases driven by hyperactivation, aberrant activation, constitutive activation, gain-of-function mutations of MEK kinases and/or substrate kinases including but not limited to ERK.
  • Such diseases encompass hyperproliferative disorders including but not limited to psoriasis, keloids, hyperplasia of the skin, benign prostatic hyperplasia (BPH), solid tumors such as respiratory tract (including but not limited to small cell and non-small cell lung cancer) , brain (including but not limited to glioma, neurofibroma, plexiform neurofibroma, medulloblastoma, ependymoma, neuroectodermal and pineal tumors), breast (including but not limited to invasive ductal carcinoma, invasive lobular carcinoma, ductal and lobular carcinoma in situ), reproductive organs (including but not limited to prostate, testicular, ovarian, endometrial, cervical, vaginal, vulvar, and uterine sarcomas) , digestive tract (including but not limited to cancers of the esophagus, colon, colorectum, stomach, gallbladder, pancreas, rectum, anus, small
  • Hyperproliferative disorders also include leukemias (including but not limited to acute lymphoblastic leukemia, acute spontaneous leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, and hairy cell leukemia), sarcomas (including but not limited to soft tissue sarcomas, bone and flesh lymphoma, lymphosarcoma, rhabdomyosarcoma) and lymphoma (including but not limited to non-Hodgkin's lymphoma, AIDS-related lymphoma, cutaneous T-cell lymphoma, Burkitt's lymphoma, Hodgkin's disease and central nervous system systemic lymphoma).
  • leukemias including but not limited to acute lymphoblastic leukemia, acute spontaneous leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, and hairy cell leukemia
  • sarcomas including but not limited to soft tissue sarcomas,
  • the present invention describes inhibitors of MEK kinases for use in certain diseases involving dysregulation of mitogen extracellular kinase activity, including but not limited to hepatomegaly, heart failure, cardiac hypertrophy, diabetes, stroke, Alzheimer's disease, cystic fibrosis, septic shock, or asthma.
  • the present invention describes inhibitors of MEK kinase for use in the treatment of diseases and disorders associated with aberrant, abnormal and/or excessive angiogenesis.
  • angiogenesis-related disorders include, but are not limited to, tumor growth and metastasis, ischemic retinal vein occlusion, diabetic retinopathy, macular degeneration, neovascular glaucoma, psoriasis, inflammation, rheumatoid arthritis, Vascular graft restenosis, restenosis and in-stent restenosis.
  • the present invention also provides a method for treating acute myeloid leukemia, comprising administering the compound of the present invention or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable Salt, hydrate or solvate, or the pharmaceutical composition of the present invention.
  • the acute myeloid leukemia is relapsed and/or refractory acute myeloid leukemia.
  • the present invention also provides a method for treating glioma, comprising administering the compound of the present invention or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable Salt, hydrate or solvate, or the pharmaceutical composition of the present invention.
  • the present invention also provides a method for treating neurofibroma, comprising administering the compound of the present invention or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt to the subject , hydrate or solvate, or the pharmaceutical composition of the present invention.
  • the neurofibroma is selected from neurofibromatosis type 1 (NF1) or plexiform neurofibroma.
  • the present invention also provides a method for treating serous carcinoma, comprising administering the compound of the present invention or its tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt to the subject , hydrate or solvate, or the pharmaceutical composition of the present invention.
  • the serous carcinoma is selected from recurrent or persistent low-grade ovarian, fallopian tube or primary peritoneal serous carcinoma.
  • Combination therapy therefore comprises the administration of at least one compound of the invention and the use of at least one other pharmaceutically active agent.
  • One or more compounds of the present invention and one or more other pharmaceutically active agents may be administered together or separately, and when administered separately, simultaneously or sequentially in any order.
  • the amounts and relative timing of administration of one or more compounds of the invention and one or more other pharmaceutically active agents will be selected to achieve the desired combined therapeutic effect. specifically:
  • the invention provides a method of treating BRAF kinase-mediated cancer comprising administering to said subject a compound of the invention in combination with a BRAF inhibitor (each optionally in a tautomer, stereoisomer, pro drug, crystal form, pharmaceutically acceptable salt, hydrate or solvate), and optionally a third therapeutic agent.
  • a BRAF inhibitor each optionally in a tautomer, stereoisomer, pro drug, crystal form, pharmaceutically acceptable salt, hydrate or solvate
  • optionally a third therapeutic agent optionally a third therapeutic agent.
  • the BRAF inhibitor is selected from the following compounds disclosed in vemurafenib, dabrafenib, encorafenib or WO 2020/011141 A1:
  • the BRAF inhibitor is selected from vemurafenib, dabrafenib, encorafenib, and the following compounds disclosed in WO 2020/011141 A1:
  • the BRAF inhibitor is selected from vemurafenib, dabrafenib, encorafenib, and the following compounds disclosed in WO 2020/011141 A1:
  • the method of treating BRAF kinase-mediated cancer does not comprise a third therapeutic agent.
  • the method of treating BRAF kinase-mediated cancer comprises a third therapeutic agent.
  • the third therapeutic agent is selected from immune checkpoint inhibitors, for example, pembrolizumab (pembrolizumab), ipilimumab (ipilimumab) and nivolumab (nivolumab), ater Pearl monoclonal antibody (atezolizumab), avelumab (avelumab), durvalumab (durvalumab), pidilzumab (pidilzumab), PDR-001 (BAP049-clone-E, disclosed in and used in WO 2017/ 019896); preferably, for example, pembrolizumab, ipilimumab and nivolumab.
  • the third therapeutic agent is selected from EGFR antibodies, for example, cetuximab (cetuximab), panitumumab (panitumumab), necitumumab (Necitumumab); preferably, For example, cetuximab.
  • the third therapeutic agent is a mitotic inhibitor, for example, a CDK4/6 inhibitor; preferably, for example, palbociclib, Rui ribociclib, abemaciclib; preferably, eg, palbociclib.
  • the BRAF kinase-mediated cancer is melanoma, brain tumors such as glioblastoma multiforme (GBM), acute myeloid leukemia (AML), lung cancer, papillary carcinoma of the thyroid, low-grade ovarian cancer, colorectal cancer, multiple myeloma, and nervous system cancer.
  • the BRAF kinase-mediated cancer is metastatic or unresectable melanoma, papillary thyroid cancer, low-grade ovarian cancer, and colorectal cancer.
  • the BRAF kinase is a BARF V600 mutant kinase.
  • the BRAF V600 mutation is BRAF V600E, BRAF V600D, BRAF V600R, BRAF V600G, and BRAF V600K. In specific embodiments, the BRAF V600 mutations are BRAF V600E and BRAF V600K. In specific embodiments, the BRAF kinase-mediated cancer is BRAF V600 mutated metastatic or unresectable melanoma. In specific embodiments, the BRAF kinase-mediated cancer is BRAF V600E or BRAF V600K mutated metastatic or unresectable melanoma. In specific embodiments, the BRAF kinase-mediated cancer is BRAF V600 mutated colorectal cancer. In specific embodiments, the BRAF kinase-mediated cancer is BRAF V600E or BRAF V600K mutated colorectal cancer.
  • the present invention also provides a method of treating NRAS or KRAS or EGFR mutated cancer comprising administering to said subject a compound of the present invention in combination with an EGFR inhibitor (each optionally in a tautomer, stereoisomer, body, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate).
  • an EGFR inhibitor each optionally in a tautomer, stereoisomer, body, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate.
  • the EGFR inhibitor is selected from the group consisting of gefitinib, erlotinib, afatinib, dacomitinib, lapatinib ( lapatinib), osimertinib, amitinib, vometinib, CO-1686, WZ4002, PD153035, PF00299804, cetuximab, panitumumab, necituzumab.
  • the NRAS-mutated cancer is NRAS-mutated non-small cell lung cancer.
  • the NRAS mutation is selected from E63K, G12V, G12R, G12A, G12D, G12S and G12C, or an increase in the copy number of the NRAS gene.
  • the present invention also provides a method of treating advanced KRAS-positive metastatic colorectal cancer, comprising administering to the subject a compound of the present invention in combination with mFOLFIRI (each optionally in a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate).
  • mFOLFIRI each optionally in a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate.
  • the present invention also provides a method of treating gastrointestinal stromal tumors, comprising administering to said subject a compound of the present invention in combination with pexidartinib (each optionally in a tautomer, stereoisomer, prodrug , crystal form, pharmaceutically acceptable salt, hydrate or solvate).
  • the present invention also provides a method of treating gastrointestinal stromal tumors, comprising administering to the subject a compound of the present invention in combination with imatinib (imatinib) (each optionally in a tautomer, stereo isomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate).
  • imatinib imatinib
  • imatinib each optionally in a tautomer, stereo isomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate.
  • the present invention also provides a method of treating non-squamous lung cancer, comprising administering to said subject a compound of the present invention in combination with carboplatin and pemetrexed (each optionally in a tautomer, stereoisomer, or body, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate).
  • the present invention also provides a method of treating biliary tract cancer, comprising administering to the subject a compound of the present invention in combination with capecitabine (each optionally in a tautomer, stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, hydrate or solvate).
  • 2,3,4-Trifluorobenzoic acid (20g, 113.6mmol) was dissolved in 60ml of concentrated sulfuric acid, the reaction solution was heated to 90°C, then concentrated sulfuric acid (12g, 122.4mmol) and concentrated nitric acid (12.8g, 132.1mmol) of the mixed solution, stirred and reacted for 5 hours, TLC monitored the completion of the reaction, cooled to room temperature, slowly added the reaction solution dropwise to ice water, extracted 3-4 times with ethyl acetate, combined the organic phases, washed with saturated brine for 2- 3 times, dried over anhydrous sodium sulfate, filtered and concentrated to obtain 24.0 g of white solid with a yield of 95.6%.
  • LC-MS (APCI): m/z 220.1 (M-1) - .
  • Step 4 Synthesis of compound 2,4-diamino-3-fluoro-5-nitrobenzoic acid methyl ester
  • Step 5 Synthesis of the compound 2,4,5-triamino-3-fluorobenzoic acid methyl ester
  • Step 6 Synthesis of compound 4-fluoro-5-amino-1H benzimidazole-6-carboxylic acid methyl ester
  • Step 7 Synthesis of compound 4-fluoro-5-amino-1-(methyl-d3)-1H-benzimidazole-6-carboxylic acid methyl ester
  • Step 8 Synthesis of the compound 5-((4-bromo-2-fluorophenyl)amino)-4-fluoro-1-(methyl-d3)-1H-benzimidazole-6-carboxylic acid methyl ester
  • Step 1 Synthesis of the compound 4-fluoro-5-amino-1-methyl-1H-benzimidazole-6-carboxylic acid methyl ester
  • Step 2 Synthesis of compound 5-((4-bromo-2-fluorophenyl)amino)-4-fluoro-1-methyl-1H-benzimidazole-6-carboxylic acid methyl ester
  • Step 2 Synthesis of the compound 2-bromo-2,2-dideuterioacetic acid benzyl ester
  • Step 3 Synthesis of compound 2-((1,3-dioxoisoindolin-2-yl)oxy)-2,2-dideuterioacetic acid benzyl ester
  • Step 1 Compound 5-((4-bromo-2-fluorophenyl)amino)-4-fluoro-1-(methyl-d 3 )-N-(2-(ethyleneoxy)ethoxy)- Synthesis of 1H-benzimidazole-6-carboxamide
  • Step 1 Compound 2-((5-((4-bromo-2-fluorophenyl)amino)-4-fluoro-1-methyl-1H-benzimidazole-6-formylamino)oxy)- Synthesis of 2,2-dideuteriobenzyl acetate
  • Cell line HT-29 (cell type: adherent; cell number/well: 3000; culture medium: RPMI-1640+10% FBS;) cultured at 37°C, 5% CO2, and 95% humidity.
  • Fetal bovine serum FBS GBICO, Cat#10099-141
  • Luminescent Cell Viability Assay Promega, Cat#G7572
  • 96-well transparent flat-bottom black wall plate Cat #3603
  • Instruments SpectraMax Multilabel Microplate Reader, MD, 2104-0010A; CO2 Incubator, Thermo Scientific, Model 3100 Series; Biological Safety Cabinet, Thermo Scientific, Model 1300 Series A2; Inverted Microscope, Olympus, CKX41SF; Refrigerator, SIEMENS , KK25E76TI.
  • Cell culture and inoculation i) Harvest the cells in the logarithmic growth phase and use a platelet counter for cell counting. Detect cell viability with trypan blue exclusion method to ensure that the cell viability is above 90%; ii) adjust the cell concentration; add 90 ⁇ L of cell suspension to the 96-well plate; iii) place the cells in the 96-well plate at 37°C, Cultivate overnight under 5% CO2, 95% humidity conditions.
  • Drug dilution and dosing i) Prepare 10-fold drug solution, the highest concentration is 100 ⁇ M, 9 concentrations, 3.16-fold dilution, add 10 ⁇ L drug solution to each well of a 96-well plate seeded with cells, and set the concentration of each drug Three duplicate wells; ii) Place the cells in the 96-well plate that have been dosed with the drug under conditions of 37° C., 5% CO 2 , and 95% humidity to continue culturing for 72 hours, and then conduct CTG analysis.
  • Cell survival rate (%) (Lum test drug-Lum culture solution control)/(Lum cell control-Lum culture solution control) ⁇ 100%.
  • the compound of the present invention was tested in the above cytotoxicity experiment, and the results showed that the compound of the present invention has stronger activity on HT-29 cells than the non-deuterated compound Binimetinib.
  • Metabolic stability is generally used to describe the speed and extent of a compound being metabolized, and is one of the main factors affecting pharmacokinetic properties. Many compounds are substrates of CYP450 enzymes and other drug-metabolizing enzymes, and liver microsomes are CYP450-rich systems. The purpose of this experiment is to combine the compounds of the present invention with human liver microsomes and/or mouse liver microsomes Incubate separately and use LC-MS/MS to detect the remaining proportion of the compound to study the in vitro stability of metabolism.
  • Phosphate buffer saline Mix 150mL of pre-prepared KH 2 PO 4 (0.5M) solution and 700mL of K 2 HPO 4 (0.5M) solution, then adjust the mixture with K 2 HPO 4 (0.5M) solution When the pH value reaches 7.4, it is used as 5-fold concentration PBS and stored at 4°C for later use. Before use, it was diluted 5 times with ultrapure water, and 3.3 mM magnesium chloride was added to obtain phosphate buffered saline PBS (100 mM).
  • NADPH regeneration system solution use 5mL of PBS to prepare NADPH solution containing 6.5mM NADP, 16.5mM G-6-P, 3U/mL G-6-PD.
  • Internal standard stop solution use acetonitrile to prepare 50ng/mL propranolol hydrochloride and 200ng/mL tolbutamide as internal standard working solution.
  • Human liver microsome solution Add 0.31mL human liver microsome (25mg/mL) into 0.961mL PBS (pH7.4) and mix well to obtain a dilution of human liver microsome with a protein concentration of 0.625mg/mL.
  • Mouse liver microsome solution take 0.31mL mouse liver microsome (25mg/mL) and add it into 0.961mL PBS (pH7.4) and mix well to obtain a dilution of mouse liver microsome with a protein concentration of 0.625mg/mL.
  • Sample working solution Prepare the compound of the present invention and non-deuterated compound powder, positive control dextromethorphan powder and omeprazole powder to 10 mM with DMSO as the sample stock solution. Then dilute with 70% acetonitrile-water to obtain 0.25mM sample working solution.
  • the termination plate was centrifuged at 5000 rpm at 4°C for 15 min. Take 200 ⁇ L of supernatant to a 96-well plate pre-added with 200 ⁇ L of ultrapure water, mix well, use LC-MS/MS for sample analysis, and inject 10 ⁇ L.
  • the LC-MS/MS system was used to detect the peak area of the test compound, dextromethorphan, omeprazole and internal standard, and the ratio of the peak area of the compound to the internal standard was calculated.
  • the peak area of the sample and the internal standard is obtained by the mass spectrometer and Analyst software, and the substrate elimination rate constant K can be obtained by plotting the remaining amount of the compound (R%) and time using the single exponential degradation model of the Graphpad prism7.0 software
  • Rats were fed with standard feed and given water. Fasting started 16 hours before the test. Drugs were dissolved with PEG400 and DMSO. Orbital blood was collected at 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours and 24 hours after administration.
  • Rats were briefly anesthetized after inhalation of ether, and 300 ⁇ L blood samples were collected from the orbits in test tubes. There is 30 ⁇ L of 1% heparin saline solution in the test tube. The test tubes were dried overnight at 60°C before use. After blood sampling at the last time point, the rats were anesthetized with ether and sacrificed.
  • the compound of the present invention has better pharmacokinetic properties in animals, and therefore has better pharmacodynamics and therapeutic effects.

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Abstract

提供了一种取代的苯并咪唑类化合物及包含该化合物的组合物及其用途，所述的取代的苯并咪唑类化合物如式(I)所示化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物。式(I)化合物可作为有效的MEK1/2抑制剂，其可通过抑制活化的MEK抑制ERK磷酸化，对多种癌症模型表现出广泛的抗肿瘤活性，同时，所述化合物联合其他抗肿瘤治疗剂可以更好地治疗多种癌症。除了抑制作用和效力外，所述化合物还显示出更好的代谢稳定性和/或药代动力学性能。

Description

取代的苯并咪唑类化合物及包含该化合物的组合物及其用途 技术领域
本发明属于医药技术领域，尤其涉及一种取代的苯并咪唑类化合物及包含该化合物的组合物及其用途。更具体而言，本发明涉及某些氘取代的5-((4-溴-2-氟苯基)氨基)-4-氟-N-(2-羟基乙氧基)-1-甲基-1H-苯并咪唑-6-甲酰胺及其衍生物的化合物及其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物。这些氘取代的化合物及其组合物可用作MEK1和MEK2蛋白的强效选择性抑制剂，且可用于治疗MEK激酶导致的疾病的用途，且这些氘取代的化合物具有更优良的ADME和药代动力学性质。
背景技术
RAS/RAF/MEK/ERK激酶途径在多于30％的人类癌症中被活化。在对生长因子、激素、细胞素等的响应中RAS GTPase(GTP酶)蛋白的活化作用刺激RAF激酶的磷酸化和活化作用。然后，这些激酶磷酸化并激活胞内蛋白激酶MEK1和MEK2，随后其磷酸化并激活其它蛋白激酶ERK1和2。这种信号途径(亦称促分裂原活化蛋白激酶(MAPK)途径或胞质级联)介导细胞对生长信号的反应。这种途径的根本功能是将细胞膜处的受体活性与控制细胞增殖、分化和存活的胞质或核靶向的修饰相联系。
这种途径的结构性活化足以诱导细胞转化。由于异常受体酪氨酸激酶活化、RAS突变或RAF突变造成的MAPK途径的失调性活化通常会在人癌症中发现，并且代表了确定异常生长控制的主要因素。在人恶性肿瘤中，RAS突变是常见的，已经在大约30％的癌症中得到了确定。GTP酶蛋白的RAS家族(使鸟苷三磷酸转变为鸟苷二磷酸的蛋白)使信号从激活的生长因子受体传递至下游胞内配对物。在由活性膜结合的RAS所补充的靶向之中，重要的靶向是丝氨酸/苏氨酸蛋白激酶的RAF家族。RAF家族由三种相关的激酶(A-、B-和C-RAF)组成，它们充当RAS的下游效应子。RAS介导的RAF活化也引发MEK1和MEK2的活化，随后将酪氨酸-185和苏氨酸-183上的ERK1和ERK2(胞外信号调节的激酶1和2)磷酸化。激活的ERK1和ERK2改变位置并在核中积聚，在核中，它们可以磷酸化各种基质，包括控制细胞生长和存活的转录因子。考虑到MAPK途 径在人癌症发展过程中的重要性，在癌症及其它增殖疾病中，将信号级联的激酶组成部分合并为调节疾病进展的潜在重要靶向。
MEK1和MKE2是磷酸化各种MAPK的苏氨酸和酪氨酸残疾的双特异性激酶的更大家族的成员。MEK1和MEK2有独特的基因编码，但它们在C端催化激酶域和大部分N端调节区域内享有高度同源性(80％)。在人癌症中还没有发现MEK1和MEK2的癌基因形式，但已经表明，MEK的结构性活化导致细胞转化。除了RAF之外，MEK也可以被其它癌基因激活。迄今为止，MEK1和MEK2的已知基质只有ERK1和ERK2。除了磷酸化酪氨酸和苏氨酸残基的独特能力之外，这种异常的基质专一性使MEK1和MEK2位于信号级联中的临界点，这会使其能将许多胞外信号整合到MAPK途径中。
RAF在许多信号级联途径中的基本作用和地位已从使用哺乳动物细胞中调控和显著抑制RAF突变的研究及对模式生物使用生化与遗传技术的研究得到证实。RAF在某些肿瘤的形成中可能具有突出作用，例如，BRAF的活化等位基因已在～70％黑素瘤、40％甲状腺乳头状癌、30％低度卵巢癌和10％结直肠癌中被识别。大多数BRAF突变见于激酶结构域，单取代(V600E)占至少80％。突变的BRAF蛋白或者通过MEK升高的激酶活性或通过激活C-RAF激活RAS/RAF/MEK/ERK激酶途径。
Binimetinib(化学名称为5-((4-溴-2-氟苯基)氨基)-4-氟-N-(2-羟基乙氧基)-1-甲基-1H-苯并咪唑-6-甲酰胺，其具有以下结构式)是Array BioPharma开发的一种强效的非ATP竞争性、高选择性的MEK1/2抑制剂，可以在纳摩尔浓度下抑制MEK、ERK磷酸化和BRAF或KRAS突变癌细胞的生长。美国食品药品监督管理局(FDA)于2013年11月授予Binimetinib孤儿药地位，并于2018年6月批准其联合BRAF抑制剂Encorafenib用于治疗具有BRAF V600E/K突变的转移性或不能切除的黑色素瘤患者。目前已证实BRAF抑制剂与MEK抑制剂联合使用可以提高疗效，并可能减少毒性作用。在美国国家综合癌症网络(National Comprehensive Cancer Network，NCCN)指南中，BRAF抑制剂/MEK抑制剂联合免疫疗法被推荐作为转移性或不可切除的黑色素瘤的一线疗法。
Figure PCTCN2022098557-appb-000001
已知较差的吸收、分布、代谢和/或排泄(ADME)性质是导致许多候选药物临床试验失败的主要原因。当前上市的许多药物也由于较差的ADME性质限制了它们的应用范围。药物的快速代谢会导致许多本来可以高效治疗疾病的药物由于过快的从体内代谢清除掉而难以成药。频繁或高剂量服药虽然有可能解决药物快速清除的问题，但该方法会带来诸如病人依从性差、高剂量服药引起的副作用及治疗成本上升等问题。另外，快速代谢的药物也可能会使患者暴露于不良的毒性或反应性代谢物中。
发现具有很好的口服生物利用度且有成药性的新型有效的高选择性MEK1/2抑制剂还是具有挑战性的工作。因此，本领域仍需开发对适用作MEK1/2抑制剂具有选择性抑制活性和/或更好地药效学/药代动力学的化合物，本发明提供了这样的化合物。
发明内容
针对以上技术问题，本发明公开了一种新型的氘取代的苯并咪唑类化合物作为有效的MEK1/2抑制剂，其可通过抑制活化的MEK抑制ERK磷酸化，对多种癌症模型表现出广泛的抗肿瘤活性，包括黑色素瘤、急性髓系白血病、神经胶质瘤、神经纤维瘤、非小细胞肺癌、乳腺癌、浆液性癌、胃肠道间质瘤、肺非鳞癌、结直肠癌、胆道癌、骨髓瘤等。同时，本发明化合物联合其他抗肿瘤治疗剂治疗多种癌症。除了抑制作用和效力外，本发明化合物还显示出良好的溶解性以及更好地代谢稳定性和/或药代动力学性能。
对此，本发明采用以下技术方案：
本发明的第一方面，提供了式(I)化合物：
Figure PCTCN2022098557-appb-000002
其中，
Y 1、Y 2、Y 3、Y 4和Y 5各自独立地选自氢、氘或卤素；
R 1、R 2、R 3和R 4各自独立地选自氢或氘；
X各自独立地选自CH 3、CD 3、CHD 2或CH 2D；
附加条件是，上述化合物至少含有一个氘原子；
或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物。
在另一方面，本发明提供了含有本发明化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物和药学上可接受的赋形剂的药物组合物。在具体实施方案中，本发明化合物以有效量提供在所述药物组合物中。在具体实施方案中，本发明化合物以治疗有效量提供。在具体实施方案中，本发明化合物以预防有效量提供。在具体实施方案中，药物组合物还包含另外的治疗剂。在具体实施方案中，另外的治疗剂选自BRAF抑制剂，EGFR抑制剂，EGFR抗体，免疫检查点抑制剂或CDK4/6抑制剂中的一种或几种。在具体实施方案中，BRAF抑制剂选自维莫非尼(vemurafenib)，达拉非尼(dabrafenib)，康奈非尼(encorafenib)，(S)-甲基-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，(S)-(甲基-d 3)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-异丙基-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，(S)-(甲基-d 3)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，(S)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d 5)氨基甲酸甲酯，(S)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7))-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d5)氨基甲酸甲酯。在具体实施方案中，EGFR抑制剂选自吉非替尼(gefitinib)，埃罗替尼(erlotinib)，阿法替尼(afatinib)，达克替尼(dacomitinib)，拉帕替尼(lapatinib)，奥希替尼(osimertinib)，阿美替尼，伏美替尼，CO-1686，WZ4002，PD153035，PF00299804。在具体实施方案中，EGFR抗体选自西妥昔单抗(cetuximab)，帕尼单抗(panitumumab)，耐昔妥珠单抗((Necitumumab)。在具体实施方案中，免疫检查点抑制剂选自帕博利珠单抗(pembrolizumab)，易普利姆妈(ipilimumab)和納武单抗(nivolumab)，阿特珠单抗(atezolizumab)，阿维鲁单抗(avelumab)，德瓦鲁单抗(durvalumab)，吡地利单抗(pidilzumab)。在具体实施方案中，CDK4/6抑制剂选自帕博西尼(palbociclib)，瑞博西尼(ribociclib)，阿贝西尼(abemaciclib)。
在另一方面，本发明提供了一种如上所述的药物组合物的制备方法，包括以下步骤：将药学上可接受的赋形剂与本发明化合物或其互变异构体、立体异构体、前药、晶型、 药学上可接受的盐、水合物或溶剂化合物进行混合，从而形成药物组合物。
在另一方面，本发明提进一步提供了治疗MEK激酶介导的疾病的方法，该方法包括向由此需要的受试者施用治疗有效量的本发明化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，或本发明药物组合物。
在具体实施方案中，所述MEK激酶介导的疾病为黑色素瘤、急性髓系白血病、神经胶质瘤、神经纤维瘤、非小细胞肺癌、乳腺癌、浆液性癌、胃肠道间质瘤、肺非鳞癌、结直肠癌、胆道癌、骨髓瘤。在具体实施方案中，所述黑色素瘤选自BRAF V600突变的黑素瘤。在具体实施方案中，所述结直肠癌选自BRAF V600突变的结直肠癌。在具体实施方案中，所述BRAF V600突变选自BRAF V600E突变或BRAF V600K突变。在具体实施方案中，所述神经纤维瘤选自神经纤维瘤病1型(NF1)或丛状神经纤维瘤；
由随后的具体实施方式、实施例和权利要求，本发明的其他目的和优点将对于本领域技术人员显而易见。
定义
本文中，如无特别说明，“氘代”指化合物或基团中的一个或多个氢被氘所取代；氘代可以是一取代、二取代、多取代或全取代。术语“一个或多个氘代的”与“一次或多次氘代”可互换使用。
本文中，如无特别说明，“非氘代的化合物”是指含氘原子比例不高于天然氘同位素含量(0.015％)的化合物。
如本文所用，术语“受试者”包括但不限于：人(即，任何年龄组的男性或女性，例如，儿科受试者(例如，婴儿、儿童、青少年)或成人受试者(例如，年轻的成人、中年的成人或年长的成人))和/或非人的动物，例如，哺乳动物，例如，灵长类(例如，食蟹猴、恒河猴)、牛、猪、马、绵羊、山羊、啮齿动物、猫和/或狗。在一些实施方案中，受试者是人。在另一些实施方案中，受试者是非人动物。
“疾病”、“障碍”和“病症”在本文中可以互换地使用。
除非另作说明，否则，本文使用的术语“治疗”包括受试者患有具体疾病、障碍或病症时所发生的作用，它降低疾病、障碍或病症的严重程度，或延迟或减缓疾病、障碍或病症的发展(“治疗性治疗”)，还包括受试者开始患有具体疾病、障碍或疾病之前发生的作用(“预防性治疗”)。
通常，化合物的“有效量”是指足以引起目标生物反应的数量。正如本领域普通技术人员所理解的那样，本发明化合物的有效量可以根据下列因素而改变：例如，生物学目标、化合物的药物动力学、所治疗的疾病、给药模式以及受试者的年龄健康情况和症状。有效量包括治疗和预防性治疗有效量。
除非另作说明，否则，本文使用的化合物的“治疗有效量”是在治疗疾病、障碍或病症的过程中足以提供治疗有益处的数量，或使与疾病、障碍或病症有关的一或多种症状延迟或最小化。化合物的治疗有效量是指单独使用或与其他疗法联用的治疗剂的数量，它在治疗疾病、障碍或病症的过程中提供治疗益处。术语“治疗有效量”可以包括改善总体治疗、降低或避免疾病或病症的症状或病因、或增强其他治疗剂的治疗效能的数量。
除非另作说明，否则，本文使用的化合物的“预防有效量”是足以预防疾病、障碍或病症的数量，或足以预防与疾病、障碍或病症有关的一或多种症状的数量，或防止疾病、障碍或病症复发的数量。化合物的预防有效量是指单独使用或与其它药剂联用的治疗剂的数量，它在预防疾病、障碍或病症的过程中提供预防益处。术语“预防有效量”可以包括改善总体预防的数量，或增强其它预防药剂的预防效能的数量。
“组合”以及相关术语是指同时或依次给药本发明的治疗剂。例如，本发明化合物可以与另一治疗剂以分开的单位剂型同时或依次给药，或与另一治疗剂一起呈单一单位剂型同时给药。
具体实施方式
化合物
本文中，“本发明化合物”指的是以下式(I)和式(II)化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物。
在一个实施方案中，本发明涉及式(I)化合物：
Figure PCTCN2022098557-appb-000003
其中，
Y 1、Y 2、Y 3、Y 4和Y 5各自独立地选自氢、氘或卤素；
R 1、R 2、R 3和R 4各自独立地选自氢或氘；
X选自CH 3、CD 3、CHD 2或CH 2D；
附加条件是，上述化合物至少含有一个氘原子；
或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物。
在一个具体的实施方案中，氘在氘代位置的氘同位素含量至少是大于天然氘同位素含量0.015％，较佳地大于30％，更佳地大于50％，更佳地大于75％，更佳地大于95％，更佳地大于99％。
具体地说，在本发明中Y 1、Y 2、Y 3、Y 4、Y 5、R 1、R 2、R 3、R 4和X的各氘代位置的氘同位素含量至少是大于天然同位素含量0.015％，更佳地大于1％，更佳地大于5％，更佳地大于10％，更佳地大于15％，更佳地大于20％，更佳地大于25％，更佳地大于30％，更佳地大于35％，更佳地大于40％，更佳地大于45％，更佳地大于50％，更佳地大于55％，更佳地大于60％，更佳地大于65％，更佳地大于70％，更佳地大于75％，更佳地大于80％，更佳地大于85％，更佳地大于90％，更佳地大于95％，更佳地大于99％。
在另一个具体的实施方案中，本发明化合物至少含有一个氘原子，更佳地含有二个氘原子，更佳地含有三个氘原子，更佳地含有四个氘原子，更佳地含有五个氘原子，更佳地含有六个氘原子，更佳地含有七个氘原子，更佳地含有八个氘原子，更佳地含有九个氘原子，更佳地含有十个氘原子，更佳地含有十一个氘原子，更佳地含有十二个氘原子。
在另一具体实施方案中，“Y 1、Y 2、Y 3、Y 4和Y 5各自独立地选自氢、氘或卤素”包括Y 1选自氢、氘或卤素，Y 2选自氢、氘或卤素，Y 3选自氢、氘或卤素，以此类推，直至Y 5选自氢、氘或卤素的技术方案。更具体地，包括Y 1是氢、Y 1是氘或Y 1是卤素(F、Cl、Br或I)，Y 2是氢、Y 2是氘或Y 2是卤素(F、Cl、Br或I)，Y 3是氢、Y 3是氘或Y 3是卤素(F、Cl、Br或I)，以此类推，直至Y 5是氢、Y 5是氘或Y 5是卤素(F、Cl、Br或I)的技术方案。
在另一具体实施方案中，“R 1、R 2、R 3和R 4各自独立地选自氢或氘”包括R 1选自氢或氘，R 2选自氢或氘，R 3选自氢或氘，和R 4选自氢或氘的技术方案。更具体地，包括 R 1是氢或R 1是氘，R 2是氢或R 2是氘，R 3是氢或R 3是氘，和R 4是氢或R 4是氘的技术方案。
在另一具体实施方案中，“X选自CH 3、CD 3、CHD 2或CH 2D”包括X是CH 3、X是CD 3、X是CHD 2或X是CH 2D的技术方案。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，Y 1、Y 2、Y 3、Y 4、Y 5、R 1、R 2、R 3和R 4如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，R 1和R 2是氘，Y 1、Y 2、Y 3、Y 4、Y 5、R 3、R 4和X如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，R 3和R 4是氘，Y 1、Y 2、Y 3、Y 4、Y 5、R 1、R 2和X如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，R 1、R 2、R 3和R 4是氘，Y 1、Y 2、Y 3、Y 4、Y 5和X如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，Y 1是氘，Y 2、Y 3、Y 4、Y 5、R 1、R 2、R 3、R 4和X如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，Y 1是氘，Y 2、Y 3、Y 4、Y 5、R 1、R 2、R 3和R 4如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，R 1和R 2是氘，Y 1、Y 2、Y 3、Y 4、Y 5、R 3和R 4如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，R 3和R 4是氘，Y 1、Y 2、Y 3、Y 4、Y 5、R 1和R 2如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，R 1、R 2、R 3和R 4是氘，Y 1、Y 2、Y 3、Y 4和Y 5如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，Y 1、R 1和R 2是氘，Y 2、Y 3、Y 4、Y 5、R 3、R 4和X如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，Y 1、R 3和R 4是氘，Y 2、Y 3、Y 4、Y 5、R 1、R 2和X如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，Y 1、R 1、R 2、R 3和R 4是氘，Y 2、Y 3、Y 4、Y 5和X如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，Y 1、R 1和R 2是氘，Y 2、Y 3、Y 4、Y 5、R 3和R 4如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，Y 1、R 3和R 4是氘，Y 2、Y 3、Y 4、Y 5、R 1和R 2如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，Y 1、R 1、R 2、R 3和R 4是氘，Y 2、Y 3、Y 4和Y 5如上文所定义。
在另一个实施方案中，本发明涉及式(II)化合物：
Figure PCTCN2022098557-appb-000004
其中，
Y 1选自氢、氘或卤素；
R 1、R 2、R 3和R 4各自独立地选自氢或氘；
X选自CH 3、CD 3、CHD 2或CH 2D；
附加条件是，上述化合物至少含有一个氘原子；
或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，Y 1、R 1、R 2、R 3和R 4如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，R 1和R 2是氘，Y 1、R 3、R 4和X如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，R 3和R 4是氘，Y 1、R 1、R 2和X如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，R 1、R 2、R 3和R 4是氘，Y 1和X如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，Y 1是氘，R 1、R 2、R 3、R 4和X如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，Y 1是氘，R 1、R 2、R 3和R 4如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，R 1和R 2是氘，Y 1、R 3、R 4和Y 5如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，R 3和R 4是氘，Y 1、R 1和R 2如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，R 1、R 2、R 3和R 4是氘，Y 1如上文所定义。
在通式(I)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，Y 1、R 1和R 2是氘，R 3、R 4和X如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，Y 1、R 3和R 4是氘，R 1、R 2和X如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，Y 1、R 1、R 2、R 3和R 4是氘，X如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，Y 1、R 1和R 2是氘，R 3和R 4如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，Y 1、R 3和R 4是氘，R 1和R 2如上文所定义。
在通式(II)一些实施方案中，优选地，本发明涉及上述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3，Y 1、R 1、R 2、R 3和R 4是氘。
作为本发明的优选实施方案，所述化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，选自如下任一化合物：
Figure PCTCN2022098557-appb-000005
Figure PCTCN2022098557-appb-000006
Figure PCTCN2022098557-appb-000007
Figure PCTCN2022098557-appb-000008
本发明化合物可包括一个或多个不对称中心，且因此可以存在多种立体异构体形式，例如，对映异构体和/或非对映异构体形式。例如，本发明化合物可为单独的对映异构体、 非对映异构体或几何异构体(例如顺式和反式异构体)，或者可为立体异构体的混合物的形式，包括外消旋体混合物和富含一种或多种立体异构体的混合物。异构体可通过本领域技术人员已知的方法从混合物中分离，所述方法包括：手性高压液相色谱法(HPLC)以及手性盐的形成和结晶；或者优选的异构体可通过不对称合成来制备。
本领域技术人员将理解，有机化合物可以与溶剂形成复合物，其在该溶剂中发生反应或从该溶剂中沉淀或结晶出来。这些复合物称为“溶剂合物”。当溶剂是水时，复合物称为“水合物”。本发明涵盖了本发明化合物的所有溶剂合物。
术语“溶剂合物”是指通常由溶剂分解反应形成的与溶剂相结合的化合物或其盐的形式。这个物理缔合可包括氢键键合。常规溶剂包括包括水、甲醇、乙醇、乙酸、DMSO、THF、乙醚等。本文所述的化合物可制备成，例如，结晶形式，且可被溶剂化。合适的溶剂合物包括药学上可接受的溶剂合物且进一步包括化学计量的溶剂合物和非化学计量的溶剂合物。在一些情况下，所述溶剂合物将能够分离，例如，当一或多个溶剂分子掺入结晶固体的晶格中时。“溶剂合物”包括溶液状态的溶剂合物和可分离的溶剂合物。代表性的溶剂合物包括水合物、乙醇合物和甲醇合物。
术语“水合物”是指与水相结合的化合物。通常，包含在化合物的水合物中的水分子数与该水合物中该化合物分子数的比率确定。因此，化合物的水合物可用例如通式R·x H 2O代表，其中R是该化合物，和x是大于0的数。给定化合物可形成超过一种水合物类型，包括，例如，单水合物(x为1)、低级水合物(x是大于0且小于1的数，例如，半水合物(R·0.5H 2O))和多水合物(x为大于1的数，例如，二水合物(R·2H 2O)和六水合物(R·6H 2O))。
本发明化合物可以是无定形或结晶形式(多晶型)。此外，本发明化合物可以以一种或多种结晶形式存在。因此，本发明在其范围内包括本发明化合物的所有无定形或结晶形式。术语“多晶型物”是指特定晶体堆积排列的化合物的结晶形式(或其盐、水合物或溶剂合物)。所有的多晶型物具有相同的元素组成。不同的结晶形式通常具有不同的X射线衍射图、红外光谱、熔点、密度、硬度、晶体形状、光电性质、稳定性和溶解度。重结晶溶剂、结晶速率、贮存温度和其他因素可导致一种结晶形式占优。化合物的各种多晶型物可在不同的条件下通过结晶制备。
本发明还包括同位素标记的化合物，它们等同于本发明化合物的那些，但一个或多个原子被原子质量或质量数不同于自然界常见的原子质量或质量数的原子所代替。可以 引入本发明化合物中的同位素的实例包括氢、碳、氮、氧、磷、硫、氟和氯的同位素，分别例如 2H、 3H、 13C、 11C、 14C、 15N、 18O、 17O、 31P、 32P、 35S、 18F和 36Cl。含有上述同位素和/或其它原子的其它同位素的本发明化合物、其前体药物和所述化合物或所述前体药物的药学上可接受的盐都属于本发明的范围。某些同位素标记的本发明化合物、例如引入放射性同位素(例如 3H和 14C)的那些可用于药物和/或底物组织分布测定。氚、即 3H和碳-14、即 14C同位素是特别优选的，因为它们容易制备和检测。进而，被更重的同位素取代，例如氘、即 2H，由于代谢稳定性更高可以提供治疗上的益处，例如延长体内半衰期或减少剂量需求，因而在有些情况下可能是优选的。同位素标记的本发明式(I)化合物及其前体药物一般可以这样制备，在进行下述流程和/或实施例与制备例所公开的工艺时，用容易得到的同位素标记的试剂代替非同位素标记的试剂。
此外，前药也包括在本发明的上下文内。本文所用的术语“前药”是指在体内通过例如在血液中水解转变成其具有医学效应的活性形式的化合物。药学上可接受的前药描述于T.Higuchi和V.Stella，Prodrugs as Novel Delivery Systems，A.C.S.Symposium Series的Vol.14，Edward B.Roche，ed.，Bioreversible Carriers in Drug Design，American Pharmaceutical Association and Pergamon Press，1987，以及D.Fleisher、S.Ramon和H.Barbra“Improved oral drug delivery：solubility limitations overcome by the use of prodrugs”，Advanced Drug Delivery Reviews(1996)19(2)115-130，每篇引入本文作为参考。
前药为任何共价键合的本发明化合物，当将这种前药给予患者时，其在体内释放母体化合物。通常通过修饰官能团来制备前药，修饰是以使得该修饰可以通过常规操作或在体内裂解产生母体化合物的方式进行的。前药包括，例如，其中羟基、氨基或巯基与任意基团键合的本发明化合物，当将其给予患者时，可以裂解形成羟基、氨基或巯基。因此，前药的代表性实例包括(但不限于)式(I)化合物的羟基、巯基和氨基官能团的乙酸酯/酰胺、甲酸酯/酰胺和苯甲酸酯/酰胺衍生物。另外，在羧酸(-COOH)的情况下，可以使用酯，例如甲酯、乙酯等。酯本身可以是有活性的和/或可以在人体体内条件下水解。合适的药学上可接受的体内可水解的酯基包括容易在人体中分解而释放母体酸或其盐的那些基团。
制备本发明化合物的方法
本发明化合物(包括其盐)可使用已知有机合成技术来制备，且可按照多种可能合 成途径中的任一种(诸如下文方案中的那些)来合成。用于制备本发明化合物的反应可在合适的溶剂中进行，有机合成领域的技术人员可容易地选择溶剂。合适的溶剂可在进行反应的温度(例如，在溶剂结冻温度至溶剂沸点温度范围内的温度)下与起始物质(反应物)、中间体或产物实质上不反应。既定反应可在一种溶剂或一种以上溶剂的混合物中进行。技术人员可依据具体反应步骤来选择用于具体反应步骤的溶剂。
本发明化合物的制备可涉及不同化学基团的保护和去除保护。本领域技术人员可容易地判定是否需要保护和去除保护以及适当保护基的选择。保护基的化学性质可参见例如Wuts和Greene，Protective Groups in Organic Synthesis，第4版，John Wiley&Sons：New Jersey，(2006)，其通过引用整体并入本文中。
本发明化合物可通过化合物的消旋混合物与光学活性的拆分剂反应形成一对非对映异构体化合物、分离非对映异构体并回收光学纯度的対映体，制备成其单个立体异构体。对映体拆分时可使用本发明化合物的非对映体衍生物进行，优先可解离的复合物(例如，结晶非对映体盐)。非对映体具有显著不同的物理性质(例如，熔点、沸点、溶解度、反应性等)，并可通过这些不相似性的优势容易地得到分离。非对映体可通过色谱，优选通过基于溶解度的差异的分离/拆分技术进行分离。然后通过不会消旋化的任何实际手段，回收光学纯对映体，连同拆分试剂。适用于从消旋混合物开始拆分得到化合物立体异构体的技术的更详细的描述可见于Jean Jacques，Andre Collet，Samue1H.Wilen，“对映体、消旋体和拆分”(“Enantiomers，Racemates and Resolutions”)，John Wiley And Sons，Inc.,1981。
可按照本领域已知任何合适的方法来监测反应。例如，可通过光谱手段(诸如核磁共振(NMR)光谱法(例如 1H或 13C)、红外(IR)光谱法、分光光度法(例如，UV-可见光)、质谱(MS))或通过色谱方法(诸如高效液相色谱法(HPLC)或薄层色谱法(TLC))来监测产物形成。
药物组合物、制剂和试剂盒
在另一方面，本发明提供了药物组合物，其包含本发明化合物(还称为“活性组分”)和药学上可接受的赋形剂。在一些实施方案中，所述药物组合物包含有效量的本发明化合物。在一些实施方案中，所述药物组合物包含治疗有效量的本发明化合物。在一些实施方案中，所述药物组合物包含预防有效量的本发明化合物。
用于本发明的药学上可接受的赋形剂是指不会破坏一起调配的化合物的药理学活性的无毒载剂、佐剂或媒剂。可以用于本发明组合物中的药学上可接受的载剂、佐剂或媒剂包括(但不限于)离子交换剂、氧化铝、硬脂酸铝、卵磷脂、血清蛋白(如人类血清白蛋白)、缓冲物质(如磷酸盐)、甘氨酸、山梨酸、山梨酸钾、饱和植物脂肪酸的偏甘油酯混合物、水、盐或电解质(如硫酸鱼精蛋白)、磷酸氢二钠、磷酸氢钾、氯化钠、锌盐、硅胶、三硅酸镁、聚乙烯吡咯烷酮、基于纤维素的物质、聚乙二醇、羧甲基纤维素钠、聚丙烯酸酯、蜡、聚乙烯-聚氧丙烯-嵌段聚合物、聚乙二醇以及羊毛脂。
本发明还包括试剂盒(例如，药物包装)。所提供的试剂盒可以包括本发明化合物、其它治疗剂，以及含有本发明化合物、其它治疗剂的第一和第二容器(例如，小瓶、安瓿瓶、瓶、注射器和/或可分散包装或其它合适的容器)。在一些实施方案中，提供的试剂盒还可以任选包括第三容器，其含有用于稀释或悬浮本发明化合物和/或其它治疗剂的药用赋形剂。在一些实施方案中，提供在第一容器和第二容器中的本发明化合物和其它治疗剂组合形成一个单位剂型。
下列制剂实施例说明可根据本发明制备的代表性的药物组合物。然而，本发明不限于下列药物组合物。
示例性的制剂1-片剂：可以将干粉形式的本发明化合物与干燥的凝胶粘合剂以约1:2的重量比混合。加入较少量的硬脂酸镁作为润滑剂。使该混合物在压片机中成型为0.3-30mg片剂(每个片剂含有0.1-10mg活性化合物)。
示例性的制剂2-片剂：可以将干粉形式的本发明化合物与干燥的凝胶粘合剂以约1:2的重量比混合。加入较少量的硬脂酸镁作为润滑剂。使该混合物在压片机中成型为30-90mg片剂(每个片剂含有10-30mg活性化合物)。
示例性的制剂3-片剂：可以将干粉形式的本发明化合物与干燥的凝胶粘合剂以约1:2的重量比混合。加入较少量的硬脂酸镁作为润滑剂。使该混合物在压片机中成型为90-150mg片剂(每个片剂含有30-50mg活性化合物)。
示例性的制剂4-片剂：可以将干粉形式的本发明化合物与干燥的凝胶粘合剂以约1:2的重量比混合。加入较少量的硬脂酸镁作为润滑剂。使该混合物在压片机中成型为150-240mg片剂(每个片剂含有50-80mg活性化合物)。
示例性的制剂5-片剂：可以将干粉形式的本发明化合物与干燥的凝胶粘合剂以约1:2的重量比混合。加入较少量的硬脂酸镁作为润滑剂。使该混合物在压片机中成型为 240-270mg片剂(每个片剂含有80-90mg活性化合物)。
示例性的制剂6-片剂：可以将干粉形式的本发明化合物与干燥的凝胶粘合剂以约1:2的重量比混合。加入较少量的硬脂酸镁作为润滑剂。使该混合物在压片机中成型为270-450mg片剂(每个片剂含有90-150mg活性化合物)。
示例性的制剂7-片剂：可以将干粉形式的本发明化合物与干燥的凝胶粘合剂以约1:2的重量比混合。加入较少量的硬脂酸镁作为润滑剂。使该混合物在压片机中成型为450-900mg片剂(每个片剂含有150-300mg活性化合物)。
示例性的制剂8-胶囊剂：可以将干粉形式的本发明化合物与淀粉稀释剂以约1:1的重量比混合。将该混合物填充到250mg胶囊中(每个胶囊含有125mg活性化合物)。
示例性的制剂9-液体：可以将本发明化合物(125mg)与蔗糖(1.75g)和黄原胶(4mg)混合，且可将得到的混合物共混，通过No.10筛目美国筛，然后与预先制备的微晶纤维素和羧甲基纤维素钠(11:89，50mg)的水溶液混合。将苯甲酸钠(10mg)、调味剂和着色剂用水稀释，并在搅拌下加入。然后，可以加入充足的水，得到5mL的总体积。
示例性的制剂10-注射剂：可以将本发明化合物溶解或悬浮在缓冲无菌盐水可注射的水性介质中，达到约5mg/mL的浓度。
给药
本发明提供的药物组合物可以通过许多途径给药，包括但不限于：口服给药、肠胃外给药、吸入给药、局部给药、直肠给药、鼻腔给药、口腔给药、阴道给药、通过植入剂给药或其它给药方式。例如，本文使用的肠胃外给药包括皮下给药、皮内给药、静脉内给药、肌肉内给药、关节内给药、动脉内给药、滑膜腔内给药、胸骨内给药、脑脊髓膜内给药、病灶内给药、和颅内的注射或输液技术。
通常，给予有效量的本文所提供的化合物。按照有关情况，包括所治疗的病症、选择的给药途径、实际给予的化合物、个体患者的年龄、体重和响应、患者症状的严重程度，等等，可以由医生确定实际上给予的化合物的量。
当用于预防本发明所述病症时，给予处于形成所述病症危险之中的受试者本文所提供的化合物，典型地基于医生的建议并在医生监督下给药，剂量水平如上所述。处于形成具体病症的危险之中的受试者，通常包括具有所述病症的家族史的受试者，或通过遗传试验或筛选确定尤其对形成所述病症敏感的那些受试者。
还可以长期给予本文所提供的药物组合物(“长期给药”)。长期给药是指在长时间内给予化合物或其药物组合物，例如，3个月、6个月、1年、2年、3年、5年等等，或者可无限期地持续给药，例如，受试者的余生。在一些实施方案中，长期给药意欲在长时间内在血液中提供所述化合物的恒定水平，例如，在治疗窗内。
可以使用各种给药方法，进一步递送本发明的药物组合物。例如，在一些实施方案中，可以推注给药药物组合物，例如，为了使化合物在血液中的浓度提高至有效水平。推注剂量取决于通过身体的活性组分的目标全身性水平，例如，肌内或皮下的推注剂量使活性组分缓慢释放，而直接递送至静脉的推注(例如，通过IV静脉滴注)能够更加快速地递送，使得活性组分在血液中的浓度快速升高至有效水平。在其它实施方案中，可以以持续输液形式给予药物组合物，例如，通过IV静脉滴注，从而在受试者身体中提供稳态浓度的活性组分。此外，在其它实施方案中，可以首先给予推注剂量的药物组合物，而后持续输液。
口服组合物可以采用散装液体溶液或混悬剂或散装粉剂形式。然而，更通常，为了便于精确地剂量给药，以单位剂量形式提供所述组合物。术语“单位剂型”是指适合作为人类患者及其它哺乳动物的单元剂量的物理离散单位，每个单位包含预定数量的、适于产生所需要的治疗效果的活性物质与合适药学赋形剂。典型的单位剂量形式包括液体组合物的预装填的、预先测量的安瓿或注射器，或者在固体组合物情况下的丸剂、片剂、胶囊剂等。在这种组合物中，所述化合物通常为较少的组分(约0.1至约50重量％，或优选约1至约40重量％)，剩余部分为对于形成所需给药形式有用的各种载体或赋形剂以及加工助剂。
对于口服剂量，代表性的方案是，每天一个至五个口服剂量，尤其是两个至四个口服剂量，典型地是三个口服剂量。使用这些剂量给药模式，每个剂量提供大约0.01至大约20mg/kg的本发明化合物，优选的剂量各自提供大约0.1至大约10mg/kg，尤其是大约1至大约5mg/kg。
为了提供与使用注射剂量类似的血液水平，或比使用注射剂量更低的血液水平，通常选择透皮剂量，数量为大约0.01至大约20％重量，优选大约0.1至大约20％重量，优选大约0.1至大约10％重量，且更优选大约0.5至大约15％重量。
从大约1至大约120小时，尤其是24至96小时，注射剂量水平在大约0.1mg/kg/小时至至少10mg/kg/小时的范围。为了获得足够的稳定状态水平，还可以给予大约0.1 mg/kg至大约10mg/kg或更多的预载推注。对于40至80kg的人类患者来说，最大总剂量不能超过大约2g/天。
适于口服给药的液体形式可包括合适的水性或非水载体以及缓冲剂、悬浮剂和分散剂、着色剂、调味剂，等等。固体形式可包括，例如，任何下列组份，或具有类似性质的化合物：粘合剂，例如，微晶纤维素、黄蓍胶或明胶；赋形剂，例如，淀粉或乳糖，崩解剂，例如，褐藻酸、Primogel或玉米淀粉；润滑剂，例如，硬脂酸镁；助流剂，例如，胶体二氧化硅；甜味剂，例如，蔗糖或糖精；或调味剂，例如，薄荷、水杨酸甲酯或橙味调味剂。
可注射的组合物典型地基于可注射用的无菌盐水或磷酸盐缓冲盐水，或本领域中已知的其它可注射的赋形剂。如前所述，在这种组合物中，活性化合物典型地为较少的组分，经常为约0.05至10％重量，剩余部分为可注射的赋形剂等。
典型地将透皮组合物配制为含有活性组分的局部软膏剂或乳膏剂。当配制为软膏剂时，活性组分典型地与石蜡或可与水混溶的软膏基质组合。或者，活性组分可与例如水包油型乳膏基质一起配制为乳膏剂。这种透皮制剂是本领域中公知的，且通常包括用于提升活性组分或制剂的稳定的皮肤渗透的其它组份。所有这种已知的透皮制剂和组份包括在本发明提供的范围内。
本发明化合物还可通过经皮装置给予。因此，经皮给药可使用贮存器(reservoir)或多孔膜类型、或者多种固体基质的贴剂实现。
用于口服给予、注射或局部给予的组合物的上述组份仅仅是代表性的。其它材料以及加工技术等阐述于Remington's Pharmaceutical Sciences,17th edition,1985,Mack Publishing Company,Easton,Pennsylvania的第8部分中，本文以引用的方式引入该文献。
本发明化合物还可以以持续释放形式给予，或从持续释放给药系统中给予。代表性的持续释放材料的描述可在Remington's Pharmaceutical Sciences中找到。
本发明还涉及本发明化合物的药学上可接受的制剂。在一个实施方案中，所述制剂包含水。在另一个实施方案中，所述制剂包含环糊精衍生物。最常见的环糊精为分别由6、7和8个α-1,4-连接的葡萄糖单元组成的α-、β-和γ-环糊精，其在连接的糖部分上任选包括一个或多个取代基，其包括但不限于：甲基化的、羟基烷基化的、酰化的和磺烷基醚取代。在一些实施方案中，所述环糊精为磺烷基醚β-环糊精，例如，磺丁基醚β-环糊精，也称作Captisol。参见，例如，U.S.5,376,645。在一些实施方案中，所述制剂包括 六丙基-β-环糊精(例如，在水中，10-50％)。
适应症
本发明提供一种治疗受试者中的疾病，如MEK激酶介导的疾病的方法，包括向所述受试者给药本发明化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，或本发明药物组合物。所述治疗方法还可以与其他疗法例如放疗、化疗相组合。
在具体实施方案中，MEK激酶介导的疾病包括炎性疾病、感染、自体免疫障碍、卒中、局部缺血、心脏障碍、神经障碍、纤维化障碍、增殖性障碍、过度增殖性障碍、肿瘤、白血病、赘生物、癌症、恶性肿瘤、代谢疾病和恶性疾病。
本发明还提供一种治疗MEK激酶介导的炎性疾病的方法，包括向所述受试者给药本发明化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，或本发明药物组合物。
在具体实施方案中，MEK激酶介导的炎性疾病包括类风湿性关节炎或多发性硬化症。
本发明还提供一种治疗MEK激酶介导的增殖性疾病的方法，包括向所述受试者给药本发明化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，或本发明药物组合物。
在具体实施方案中，MEK激酶介导的增殖性疾病包括癌症、银屑病、再狭窄、自体免疫疾病或动脉粥样硬化。
本发明还提供一种治疗MEK激酶介导的癌症的方法，包括向所述受试者给药本发明化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，或本发明药物组合物。
在具体实施方案中，MEK激酶介导的癌症包括黑色素瘤(例如，BRAF V600突变的黑素瘤)、急性髓系白血病、神经胶质流、神经纤维瘤(例如，神经纤维瘤病1型(NF1)或丛状神经纤维瘤)、非小细胞肺癌、乳腺癌、浆液性癌、胃肠道间质瘤、肺非鳞癌、结直肠癌(例如，BRAF V600突变的结直肠癌)、胆道癌、骨髓瘤。
本发明描述了MEK激酶的抑制剂，其用于由MEK激酶和/或底物激酶包括但不限于ERK的过度激活、异常激活、组成性激活、获得功能的突变所驱动的疾病的治疗。 这样的疾病涵盖过度增殖性疾病，其包括但不限于银屑病、瘢痕瘤、皮肤的增生、良性前列腺瘤增生(BPH)、实体肿瘤例如呼吸道(包括但不限于小细胞和非小细胞肺癌)、脑(包括但不限于神经胶质瘤、神经纤维瘤、丛状神经纤维瘤、成神经管细胞瘤、室管膜瘤、神经外胚层和松果体肿瘤)、乳腺(包括但不限于侵入性导管癌、侵入性小叶癌、原位导管和小叶癌)、生殖器官(包括但不限于前列腺癌、睾丸癌、卵巢癌、子宫内膜癌、宫颈癌、阴道癌、外阴癌和子宫肉瘤)、消化道(包括但不限于食管、结肠、结肠直肠、胃、胆囊、胰腺、直肠、肛门、小肠和唾液腺癌)、尿道(包括但不限于膀胱、输尿管、肾、肾脏、尿道和肾乳头状癌)、眼(包括但不限于眼内黑素瘤和成视网膜细胞瘤)、肝(包括但不限于肝细胞癌和胆管癌)、皮肤(包括但不限于黑素瘤、鳞状细胞癌、卡波斯肉瘤、Merkel细胞皮肤癌、非黑素瘤皮肤癌)、头颈部(包括但不限于喉、鼻咽、下咽、口咽癌、唇和口腔癌和鳞状细胞癌)、甲状腺、甲状旁腺的癌症和它们的转移肿瘤。过度增殖性疾病还包括白血病(包括但不限于急性成淋巴细胞性白血病、急性随性白血病、慢性髓性白血病、慢性淋巴细胞性白血病和毛细胞白血病)、肉瘤(包括但不限于软组织肉瘤、骨肉瘤、淋巴肉瘤、横纹肌肉瘤)和淋巴瘤(包括但不限于非霍奇金淋巴瘤、AIDS相关的淋巴瘤、皮肤T细胞淋巴瘤、伯基特氏淋巴瘤、霍奇金氏病和中枢神经系统的淋巴瘤)。
本发明描述了MEK激酶的抑制剂，其用于某些涉及有丝分裂原细胞外激酶活性的异常调控的疾病，包括但不限于肝肿大、心力衰竭、心脏肥大、糖尿病、卒中、阿兹海默氏病、囊性纤维化、脓毒性休克或哮喘。
本发明描述了MEK激酶的抑制剂，其用于治疗与畸变、异常和/或过度的血管发生相关的疾病和障碍。这样的与血管发生相关的障碍包括但不限于肿瘤生长和转移、局部缺血性视网膜静脉阻塞、糖尿病性视网膜病、黄斑变性、新生血管性青光眼、银屑病、炎症、类风湿性关节炎、血管移植物再狭窄、再狭窄和支架内再狭窄。
本发明还提供一种治疗急性髓系白血病的方法，包括向所述受试者给药本发明化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，或本发明药物组合物。在具体实施方案中，所述急性髓系白血病为复发性和/或难治性急性髓系白血病。
本发明还提供一种治疗神经胶质瘤的方法，包括向所述受试者给药本发明化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，或本发明药物组合物。
本发明还提供一种治疗神经纤维瘤的方法，包括向所述受试者给药本发明化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，或本发明药物组合物。在具体实施方案中，所述神经纤维瘤选自神经纤维瘤病1型(NF1)或丛状神经纤维瘤。
本发明还提供一种治疗浆液性癌的方法，包括向所述受试者给药本发明化合物或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，或本发明药物组合物。在具体实施方案中，所述浆液性癌选自复发或持续低度卵巢、输卵管或原发性腹膜浆液性癌。
组合疗法
本发明化合物可以单独使用或与其他治疗剂组合使用。根据本发明的组合治疗因此包括给予至少一种本发明化合物和使用至少一种其他的药学活性剂。一种或多种本发明化合物和一种或多种其他药学活性剂可以一起给药或分开给药，当分开给药时，可以同时进行或以任何顺序相继进行。将选择一种或多种本发明化合物和一种或多种其他药学活性剂的量和相对给药时机以实现期望的组合治疗效果。具体地：
本发明提供一种治疗BRAF激酶介导的癌症的方法，包括向所述受试者组合给药本发明化合物和BRAF抑制剂(各自任选地处于互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物)，和任选地第三治疗剂。
在具体实施方案中，BRAF抑制剂选自维莫非尼(vemurafenib)，达拉非尼(dabrafenib)，康奈非尼(encorafenib)或WO 2020/011141 A1公开的如下化合物：
Figure PCTCN2022098557-appb-000009
WO 2020/011141 A1中公开的化合物
优选地，BRAF抑制剂选自维莫非尼(vemurafenib)，达拉非尼(dabrafenib)，康奈非尼(encorafenib)，和WO 2020/011141 A1公开的如下化合物：
(S)-甲基-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，
(S)-(甲基-d 3)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-异丙基-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，
(S)-(甲基-d 3)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，
(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d 5)氨基甲酸甲酯，
(S)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d 5)氨基甲酸甲酯，
(R)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d 5)氨基甲酸甲酯，
(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7))-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d5)氨基甲酸甲酯，
(S)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7))-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d 5)氨基甲酸甲酯，
(R)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7))-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d5)氨基甲酸甲酯。
更优选地，BRAF抑制剂选自维莫非尼(vemurafenib)，达拉非尼(dabrafenib)，康奈非尼(encorafenib)，和WO 2020/011141 A1公开的如下化合物：
(S)-甲基-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，
(S)-(甲基-d 3)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-异丙基-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，
(S)-(甲基-d 3)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，
(S)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d 5)氨基甲酸甲酯，
(S)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7))-1H-吡唑-4-基)嘧啶- 2-基)氨基)丙-2-基-1,1,3,3,3-d 5)氨基甲酸甲酯。
在具体实施方案中，所述治疗BRAF激酶介导的癌症的方法不包含第三治疗剂。
在具体实施方案中，所述治疗BRAF激酶介导的癌症的方法包含第三治疗剂。在具体实施方案中，所述第三治疗剂选自免疫检查点抑制剂，例如，帕博利珠单抗(pembrolizumab)，易普利姆妈(ipilimumab)和納武单抗(nivolumab)，阿特珠单抗(atezolizumab)，阿维鲁单抗(avelumab)，德瓦鲁单抗(durvalumab)，吡地利单抗(pidilzumab)、PDR-001(BAP049-克隆-E，公开于并用于WO 2017/019896中)；优选地，例如，帕博利珠单抗(pembrolizumab)，易普利姆妈(ipilimumab)和納武单抗(nivolumab)。在具体实施方案中，所述第三治疗剂选自EGFR抗体，例如，西妥昔单抗(cetuximab)，帕尼单抗(panitumumab)，耐昔妥珠单抗((Necitumumab)；优选地，例如，西妥昔单抗(cetuximab)。在具体实施方案中，所述第三治疗剂是有丝分裂抑制剂，例如，CDK4/6抑制剂；优选的，例如，帕博西尼(palbociclib)，瑞博西尼(ribociclib)，阿贝西尼(abemaciclib)；优选地，例如，帕博西尼(palbociclib)。
在具体实施方案中，BRAF激酶介导的癌症是黑素瘤、脑瘤例如多形性成胶质细胞瘤(GBM)、急性髓性白血病(AML)、肺癌、甲状腺乳头状癌、低度卵巢癌、结直肠癌、多发性骨髓瘤和神经系统癌。优选地，BRAF激酶介导的癌症是转移性或不可切除的黑素瘤、甲状腺乳头状癌、低度卵巢癌和结直肠癌。在具体的实施方案中，BRAF激酶是BARF V600突变激酶。在具体实施方案中，BRAF V600突变是BRAF V600E、BRAF V600D、BRAF V600R、BRAF V600G和BRAF V600K。在具体实施方案中，BRAF V600突变是BRAF V600E和BRAF V600K。在具体的实施方案中，BRAF激酶介导的癌症是BRAF V600突变的转移性或不可切除的黑素瘤。在具体的实施方案中，BRAF激酶介导的癌症是BRAF V600E或BRAF V600K突变的转移性或不可切除的黑素瘤。在具体的实施方案中，BRAF激酶介导的癌症是BRAF V600突变的结直肠癌。在具体的实施方案中，BRAF激酶介导的癌症是BRAF V600E或BRAF V600K突变的结直肠癌。
本发明还提供一种治疗NRAS或KRAS或EGFR突变的癌症的方法，包括向所述受试者组合给药本发明化合物和EGFR抑制剂(各自任选地处于互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物)。在具体实施方案中，所述EGFR抑制剂选自吉非替尼(gefitinib)、埃罗替尼(erlotinib)、阿法替尼(afatinib)、达克替 尼(dacomitinib)、拉帕替尼(lapatinib)、奥希替尼(osimertinib)、阿美替尼、伏美替尼、CO-1686、WZ4002、PD153035、PF00299804、西妥昔单抗、帕尼单抗、耐昔妥珠单抗。在具体实施方案中，NRAS突变的癌症是NRAS突变的非小细胞肺癌。在具体的实施方案中，所述NRAS突变选自E63K、G12V、G12R、G12A、G12D、G12S和G12C，或NRAS基因拷贝数的增加。
本发明还提供一种治疗晚期KRAS阳性转移性结直肠癌的方法，包括向所述受试者组合给药本发明化合物和mFOLFIRI(各自任选地处于互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物)。
本发明还提供一种治疗胃肠道间质瘤的方法，包括向所述受试者组合给药本发明化合物和pexidartinib(各自任选地处于互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物)。
本发明还提供一种治疗胃肠道间质瘤的方法，包括向所述受试者组合给药本发明化合物和伊马替尼(imatinib)(各自任选地处于互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物)。
本发明还提供一种治疗肺非鳞癌的方法，包括向所述受试者组合给药本发明化合物联合卡铂和培美曲塞(各自任选地处于互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物)。
本发明还提供一种治疗胆道癌的方法，包括向所述受试者组合给药本发明化合物联合卡培他滨(capecitabine)(各自任选地处于互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物)。
实施例
下面结合具体实施例，作进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。除非另外说明，否则份数和百分比为重量份和重量百分比。
缩略语：
Pd 2(dba) 3：三(二亚苄基丙酮)二钯
Xant-phos:4,5-双二苯基膦-9,9-二甲基氧杂蒽
NBS：N-溴代琥珀酰亚胺
NIS：N-碘代琥珀酰亚胺
PTSA：对甲苯磺酸
EDCI：1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐
HOBT：1-羟基苯并三氮唑
DMAP：二甲氨基吡啶
TEA：三乙胺
DIEA：N,N-二异丙基乙胺
TFAA：三氟乙酸酐
Na 2CO 3：碳酸钠
K 2CO 3：碳酸钾
Cs 2CO 3：碳酸铯
EtOH：乙醇
BnOH：苄醇
DCM：二氯甲烷
THF：四氢呋喃
ACN：乙腈
DME：乙二醇二甲醚
DMF：N,N-二甲基甲酰胺
DMSO：二甲基亚砜
TMSCl：三甲基氯硅烷
中间体A-1 5-((4-溴-2-氟苯基)氨基)-4-氟-1-(甲基-d 3)-1H-苯并咪唑-6-甲酸制备
Figure PCTCN2022098557-appb-000010
采用以下合成路线
Figure PCTCN2022098557-appb-000011
步骤1：化合物2,3,4-三氟-5-硝基苯甲酸的合成
将2,3,4-三氟苯甲酸(20g，113.6mmol)溶于60ml浓硫酸中，将反应液加热至90℃，然后滴加浓硫酸(12g，122.4mmol)和浓硝酸(12.8g，132.1mmol)的混合溶液，搅拌反应5小时，TLC监测反应完毕，冷却至室温，将反应液缓慢滴加入冰水中，用乙酸乙酯萃取3-4次，合并有机相，饱和食盐水洗涤2-3次，无水硫酸钠干燥，过滤浓缩后得白色固体24.0g，收率95.6％。LC-MS(APCI):m/z＝220.1(M-1) -。
步骤2：化合物2,3-二氟-4-氨基-5-硝基苯甲酸的合成
将上步所得2,3,4-三氟-5-硝基苯甲酸(5.81g，26.3mmol)置于100ml烧瓶中，加入30ml去离子水，将混合物冰浴下降温至0℃，缓慢滴加浓氨水(18.4g，131.5mmol)，加毕，升至室温搅拌反应过夜，TLC监测反应完毕，冰浴下用1N稀盐酸调pH＜2，析出淡黄色固体，过滤后真空干燥得到4.99g产物，收率87.1％。LC-MS(APCI):m/z＝219.4(M+1) +。
步骤3：化合物2,3-二氟-4-氨基-5-硝基苯甲酸甲酯的合成
在反应瓶中加入2,3-二氟-4-氨基-5-硝基苯甲酸(4.99g，22.9mmol)和三甲基氯硅烷(4.97g，45.8mmol)，加入100ml甲醇溶解，氮气保护下将反应液加热至65℃搅拌过夜，TLC监测反应完毕，冷却至室温，浓缩后经硅胶柱分离得黄色固体4.51g，收率84.9％。LC-MS(APCI):m/z＝233.4(M+1) +。
步骤4：化合物2,4-二氨基-3-氟-5-硝基苯甲酸甲酯的合成
将上步所得2,3-二氟-4-氨基-5-硝基苯甲酸甲酯(1.0g，4.3mmol)溶于10ml二氧 六环中，加入氨水(1.6ml，21.4mmol)，氮气保护下升温至90℃反应2-4小时，TLC监测反应完毕。将反应液冷却至室温，加入30ml水稀释，用乙酸乙酯萃取3-4次，合并有机相，饱和食盐水洗涤，无水硫酸钠干燥，过滤，滤液浓缩后干燥得到1.03g黄色固体，直接投入到下一步反应。LC-MS(APCI):m/z＝230.8(M+1) +.
步骤5：化合物2,4,5-三氨基-3-氟基苯甲酸甲酯的合成
将上步所得2,4-二氨基-3-氟-5-硝基苯甲酸甲酯(1.03g，4.5mmol)、还原铁粉(2.5g，45mmol)和无水氯化铵(1.44g，26.9mmol)置于100ml烧瓶中，加入15ml乙醇和5ml水，加热至70℃搅拌反应1-2小时，TLC监测反应完毕。硅藻土助滤除去催化剂，滤液浓缩后得灰色固体860mg，无需纯化直接投入到下一步反应。LC-MS(APCI):m/z＝200.1(M+1) +。
步骤6：化合物4-氟-5-氨基-1H苯并咪唑-6-甲酸甲酯的合成
将上步所得2,4,5-三氨基-3-氟基苯甲酸甲酯(860mg，4.32mmol)和乙酸甲脒(542mg，5.21mmol)加入到10ml无水乙醇中，升温至80℃回流反应4小时，TLC监测反应完毕。将反应液冷却至室温，浓缩除去溶剂，加入20ml水稀释，用二氯甲烷萃取3-4次，合并有机相，浓缩后硅胶柱层析纯化得到486mg淡黄色固体，收率：53.8％。LC-MS(APCI):m/z＝210.5(M+1) +。
步骤7：化合物4-氟-5-氨基-1-(甲基-d3)-1H-苯并咪唑-6-甲酸甲酯的合成
将上步所得4-氟-5-氨基-1H苯并咪唑-6-甲酸甲酯(600mg，2.86mmol)、氘代碘甲烷(458mg，3.16mmol)和碳酸钾(794mg，5.76mmol)加入到50ml反应瓶中，加入10ml无水DMF，氮气保护下升温至70℃搅拌反应2小时，TLC监测反应完毕。降至室温后加入30ml水稀释，用乙酸乙酯萃取3-4次，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥，浓缩，经硅胶柱分离得棕色固体201mg，收率31.5％。LC-MS(APCI):m/z＝227.5(M+1) +。
步骤8：化合物5-((4-溴-2-氟苯基)氨基)-4-氟-1-(甲基-d3)-1H-苯并咪唑-6-甲酸甲酯的合成
将上步所得4-氟-5-氨基-1-(甲基-d3)-1H-苯并咪唑-6-甲酸甲酯(200mg，0.9mmol)、2-氟-4-溴-1-碘苯(300mg，0.99mmol)、Pd 2(dba) 3(16mg，0.017mmol)、Xantphos(26mg，0.044mmol)和碳酸铯(584mg，1.79mmol)加入到20ml微波管中，氮气保护下加入8ml乙二醇二甲醚，密封后微波加热至90℃反应1小时，TLC监测反应完毕，降至室温后浓缩除去溶剂，硅胶柱层析纯化得到229mg土黄色固体，收率：64.0％。LC-MS(APCI):m/z＝399.2(M+1) +。
步骤9：中间体A-1的合成
将5-((4-溴-2-氟苯基)氨基)-4-氟-1-(甲基-d3)-1H-苯并咪唑-6-甲酸甲酯(1.63g，4.09mmol)加入到100ml反应瓶中，加入30ml四氢呋喃和10ml水溶解，再加入氢氧化钠(0.68g，17.0mmol)，加热至45℃搅拌反应3-5小时，TLC监测反应完毕后，浓缩除去四氢呋喃，加入10ml水稀释，用1N稀盐酸调pH至酸性，析出类白色固体，过滤后真空干燥得到1.3g粗品，直接投入到下一步反应。LC-MS(APCI):m/z＝385.2(M+1) +。
中间体A-2 5-((4-溴-2-氟苯基)氨基)-4-氟-1-甲基-1H-苯并咪唑-6-甲酸甲酯制备
Figure PCTCN2022098557-appb-000012
采用以下合成路线
Figure PCTCN2022098557-appb-000013
步骤1：化合物4-氟-5-氨基-1-甲基-1H-苯并咪唑-6-甲酸甲酯的合成
将4-氟-5-氨基-1H-苯并咪唑-6-羧酸甲酯(600mg，2.86mmol)、碘甲烷(453mg，3.16mmol)和碳酸钾(794mg，5.76mmol)加入到50ml反应瓶中，加入10ml无水DMF，氮气保护下升温至70℃搅拌反应2小时，TLC监测反应完毕。降至室温后加入30ml水稀释，用乙酸乙酯萃取3-4次，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥，浓缩，经硅胶柱分离得棕色固体203mg，收率31.9％。LC-MS(APCI):m/z＝224.5(M+1) +。
步骤2：化合物5-((4-溴-2-氟苯基)氨基)-4-氟-1-甲基-1H-苯并咪唑-6-甲酸甲酯的合成
将上步所得4-氟-5-氨基-1-甲基-1H-苯并咪唑-6-甲酸甲酯(200mg，0.9mmol)、2-氟-4-溴-1-碘苯(300mg，0.99mmol)、Pd2(dba)3(16mg，0.017mmol)、Xantphos(26mg，0.044mmol)和碳酸铯(584mg，1.79mmol)加入到20ml微波管中，氮气保护下加入8ml乙二醇二甲醚，密封后微波加热至90℃反应1小时，TLC监测反应完毕，降至室温后浓缩除去溶剂，硅胶柱层析纯化得到208mg土黄色固体，收率：58.1％。LC-MS(APCI):m/z＝396.2(M+1) +。
步骤3：中间体A-2的合成
将5-((4-溴-2-氟苯基)氨基)-4-氟-1-甲基-1H-苯并咪唑-6-甲酸甲酯(1.63g，4.09mmol)加入到100ml反应瓶中，加入30ml四氢呋喃和10ml水溶解，再加入氢氧化钠(0.68g，17.0mmol)，加热至45℃搅拌反应3-5小时，TLC监测反应完毕后，浓缩除去四氢呋喃，加入10ml水稀释，用1N稀盐酸调PH至酸性，析出类白色固体，过滤后真空干燥得到1.3g粗品，直接投入到下一步反应。LC-MS(APCI):m/z＝382.1(M+1) +。
中间体B-1 2-(氨基氧基)-2,2-二氘代乙酸苄酯制备
Figure PCTCN2022098557-appb-000014
采用以下合成路线
Figure PCTCN2022098557-appb-000015
步骤1：化合物2-溴-2,2-二氘代乙酸的合成
将氘代乙酸(10g，156mmol)加入到80ml三氟乙酸酐中，室温下加入溴的二氧六环络合物(37g，149mmol)，氮气保护下搅拌反应过夜，GC监测反应完毕，减压浓缩后得到10.8g油状液体，不需纯化直接投入到下一步反应。
步骤2：化合物2-溴-2,2-二氘代乙酸苄酯的合成
将2-溴-2,2-二氘代乙酸(10.8g，78.55mmol)、苄醇(8.5g，78.6mmol)、EDCI(16.6 g，86.6mmol)和DMAP(1.06g，8.7mmol)溶于80ml无水DMF中，氮气保护下室温搅拌反应3-5小时，TLC监测反应完毕。加入200ml水稀释，用乙酸乙酯萃取3-4次，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥，浓缩，经硅胶柱分离得无水油状液体7.5g，两步收率21％。LC-MS(APCI):m/z＝280.3(M+1) +。
步骤3：化合物2-((1,3-二氧代异吲哚啉-2-基)氧基)-2,2-二氘代乙酸苄酯的合成
将上步所得2-溴-2,2-二氘代乙酸苄酯(3.94g，17.3mmol)，N-羟基邻苯二甲酰亚胺(2.51g，15.38mmol)和三乙胺(2.36g，23.3mmol)加入到40ml无水DMF中，氮气保护下室温搅拌过夜。加入150ml水稀释，用乙酸乙酯萃取3-4次，合并有机相，用饱和食盐水洗涤，无水硫酸钠干燥，浓缩，经硅胶柱分离得白色固体4.36g，收率81％。LC-MS(APCI):m/z＝314.4(M+1) +。
步骤4：中间体B-1的合成
将上步所得2-((1,3-二氧代异吲哚啉-2-基)氧基)-2,2-二氘代乙酸苄酯(4.36g，14.0mmol)和水合肼(1.0g，20.0mmol)溶于45ml二氯甲烷中，室温下搅拌反应过夜，TLC监测反应完毕。过滤除去不溶物，滤液浓缩后经硅胶柱分离得到黄色油状液体1.99g，收率78.3％。LC-MS(APCI):m/z＝184.7(M+1) +。
实施例1 5-((4-溴-2-氟苯基)氨基)-4-氟-N-(2-羟基乙氧基)-1-(甲基-d 3)-1H-苯并咪唑- 6-甲酰胺(化合物T-1)制备
Figure PCTCN2022098557-appb-000016
采用以下合成路线
Figure PCTCN2022098557-appb-000017
步骤1：化合物5-((4-溴-2-氟苯基)氨基)-4-氟-1-(甲基-d 3)-N-(2-(乙烯氧基)乙氧基)-1H-苯并咪唑-6-甲酰胺的合成
将中间体A-1(1.3g，3.4mmol)、O-(2-(乙烯氧基)乙基)羟胺(0.41g，4.02mmol)、EDCI(0.78g，4.06mmol)、HOBT(0.55g，4.08mmol)和三乙胺(0.41g，4.06mmol)加入到100ml反应瓶中，氮气保护下加入15ml无水DMF，室温下搅拌反应过夜，TLC监测反应完毕，加入50ml水稀释，用乙酸乙酯萃取3-4次，合并有机相，饱和食盐水洗涤，浓缩后硅胶柱层析纯化得到0.88g类白色固体，收率：55.3％。LC-MS(APCI):m/z＝470.5(M+1) +。 1H NMR(400MHz,DMSO-d 6):δ11.79(s,1H),8.39(s,1H),7.92(s,1H),7.71(d,J＝13.2Hz,1H),7.61(t,J＝9.4Hz,1H),7.23-7.27(m,1H),6.52-6.43(m,1H),6.35(dd,J＝8.7,4.1Hz,1H),4.23-4.10(m,1H),4.06-3.91(m,3H),3.83(s,2H).
步骤2：化合物T-1的合成
将上步所得5-((4-溴-2-氟苯基)氨基)-4-氟-1-(甲基-d 3)-N-(2-(乙烯氧基)乙氧基)-1H-苯并咪唑-6-甲酰胺(0.88g，1.88mmol)溶于15ml乙醇中，冰浴下降温至0℃，缓慢滴加2N稀盐酸(9.4ml，18.8mmol)，加毕，升至室温搅拌反应2-4小时，TLC监测反应完毕，用1N氢氧化钠水溶液调pH至弱碱性，浓缩除去溶剂，硅胶柱层析纯化得到0.42g白色固体，收率：50.5％。LC-MS(APCI):m/z＝444.7(M+1) +。 1H NMR(400MHz,CD 3OD):δ8.30(s,1H),8.09(s,1H),7.72(s,1H),7.50(d,J＝2.0Hz,1H),7.21-7.17(m,1H),6.40(dd,J＝8.8,4.8Hz,1H),3.97-3.89(m,2H),3.70-3.66(m,2H).
实施例2 5-((4-溴-2-氟苯基)氨基)-4-氟-N-(2-羟基乙氧基-1,1,2,2-d 4)-1-甲基-1H-苯并 咪唑-6-甲酰胺(化合物T-2)制备
Figure PCTCN2022098557-appb-000018
采用以下合成路线
Figure PCTCN2022098557-appb-000019
步骤1：化合物2-((5-((4-溴-2-氟苯基)氨基)-4-氟-1-甲基-1H-苯并咪唑-6-甲酰基氨基)氧基)-2,2-二氘代乙酸苄酯的合成
将中间体A-2(1.3g，3.4mmol)、中间体B-1(0.74g，4.02mmol)、EDCI(0.78g，4.06mmol)、HOBT(0.55g，4.08mmol)和三乙胺(0.41g，4.06mmol)加入到100ml反应瓶中，氮气保护下加入15ml无水DMF，室温下搅拌反应过夜，TLC监测反应完毕，加入50ml水稀释，用乙酸乙酯萃取3-4次，合并有机相，饱和食盐水洗涤，浓缩后硅胶柱层析纯化得到1.78g类白色固体，收率：95.7％。LC-MS(APCI):m/z＝547.3(M+1) +。 1H NMR(400MHz,DMSO-d 6):δ8.39(s,1H),7.84(s,1H),7.78-7.64(m,1H),7.58(d,J＝2.3Hz,1H),7.40-7.28(m,5H),7.23(dd,J＝8.8,2.2Hz,1H),6.32(dd,J＝8.7,3.6Hz,1H),5.13(s,2H),3.88(s,3H).
步骤2：化合物T-2的合成
将上步所得的2-((5-((4-溴-2-氟苯基)氨基)-4-氟-1-甲基-1H-苯并咪唑-6-甲酰基氨基)氧基)-2,2-二氘代乙酸苄酯(1.78g，3.25mmol)溶于40ml无水THF中，氮气保护下降温至-10℃，分批加入氘代氢化锂铝(273mg，6.5mmol)，低温下继续搅拌反应3-4小时，TLC监测反应完毕，低温下加入5ml水淬灭反应，再加入10ml 15％的氢氧化钠水溶液，硅藻土助滤，滤液浓缩后硅胶柱层析纯化得到类白色固体0.47g，收率：32.6％。LC-MS(APCI):m/z＝445.6(M+1) +。 1H NMR(400MHz,CD 3OD):δ11.73-11.64(m,1H),8.40(s,1H),7.93(brs,1H),7.74(s,1H),7.61(s,1H),7.26(dd,J＝8.8Hz,2.0Hz,1H),6.36(dd,J＝8.7Hz,4.0Hz,1H),4.67(brs,1H),3.91(s,3H).
生物活性测试。
(1)细胞毒性实验
检测实施例化合物对HT-29细胞活性的抑制效应。
细胞系：HT-29(细胞类型：贴壁；细胞数量/孔：3000；培养基：RPMI-1640+10％FBS；)置于37℃、5％CO2、95％湿度条件下培养。
耗材及试剂：胎牛血清FBS(GBICO,Cat#10099-141)、
Figure PCTCN2022098557-appb-000020
Luminescent  Cell Viability Assay(Promega,Cat#G7572)、96孔透明平底黑壁板(
Figure PCTCN2022098557-appb-000021
Cat#3603)。
仪器：SpectraMax多标记微孔板检测仪，MD，2104-0010A；CO2培养箱，Thermo Scientific，Model 3100 Series；生物安全柜，Thermo Scientific，Model 1300 Series A2；倒置显微镜，Olympus，CKX41SF；冰箱，SIEMENS，KK25E76TI。
实验步骤：
1)细胞培养和接种：i)收获处于对数生长期的细胞并采用血小板计数器进行细胞计数。用台盼蓝排斥法检测细胞活力，确保细胞活力在90％以上；ii)调整细胞浓度；分别添加90μL细胞悬液至96孔板中；iii)将96孔板中的细胞置于37℃、5％CO2、95％湿度条件下培养过夜。
2)药物稀释和加药：i)配制10倍药物溶液，最高浓度为100μM，9个浓度，3.16倍稀释，在接种有细胞的96孔板中每孔加入10μL药物溶液，每个药物浓度设置三个复孔；ii)将已加药的96孔板中的细胞置于37℃、5％CO2、95％湿度条件下继续培养72小时，之后进行CTG分析。
3)终点读板：i)融化CTG试剂并平衡细胞板至室温30分钟；ii)每孔加入等体积的CTG溶液；iii)在定轨摇床上振动5分钟使细胞裂解；iv)将细胞板放置于室温20分钟以稳定冷光信号；v)读取冷光值。
数据处理：使用GraphPad Prism 5.0软件分析数据，利用非线性S曲线回归来拟合数据得出剂量-效应曲线，并由此计算IC50值。细胞存活率(％)＝(Lum待测药-Lum培养液对照)/(Lum细胞对照-Lum培养液对照)×100％。
在上述细胞毒性实验中测试了本发明化合物，结果表明：与未经氘代化合物Binimetinib相比，本发明化合物对HT-29细胞具有更强效的活性。
(2)代谢稳定性评价
代谢稳定性一般用来描述化合物被代谢的速度和程度，是影响药代动力学性质的主要因素之一。很多化合物都是CYP450酶和其它的药物代谢酶的底物，而肝微粒体是富含CYP450的体系，本实验的目的是通过将本发明化合物与人肝微粒体和/或小鼠肝微粒体分别孵育并运用LC-MS/MS检测化合物的剩余比例来进行体代谢外稳定性的研究。
①溶液的配制
磷酸盐缓冲液(PBS)：取预先配好的KH 2PO 4(0.5M)溶液150mL和K 2HPO 4(0.5M) 溶液700mL混合，再用K 2HPO 4(0.5M)溶液调节混合液pH值至7.4，作为5倍浓度PBS，存于4℃备用。使用前用超纯水稀释5倍，加入3.3mM氯化镁，得到磷酸盐缓冲液PBS(100mM)。
NADPH再生系统溶液：用5mL的PBS配制含有6.5mM NADP，16.5mM G-6-P，3U/mL G-6-P D的NADPH溶液。
内标终止液：用乙腈配制50ng/mL盐酸普萘洛尔和200ng/mL甲苯磺丁脲作为内标工作液。
人肝微粒体溶液：取0.31mL人肝微粒体(25mg/mL)加入0.961mL PBS(pH7.4)中混匀，得到蛋白浓度为0.625mg/mL的人肝微粒体稀释液。
小鼠肝微粒体溶液：取0.31mL小鼠肝微粒体(25mg/mL)加入0.961mL PBS(pH7.4)中混匀，得到蛋白浓度为0.625mg/mL的小鼠肝微粒体稀释液。
样品工作液：用DMSO配制本发明化合物和非氘代化合物粉末、阳性对照右美沙芬粉末和奥美拉唑粉末至10mM，作为样品储备液。再用70％乙腈-水稀释得到0.25mM样品工作液。
②样品孵育
取398μL的人肝微粒体稀释液加入96孔孵育板中(N＝2)，分别加入2μL 0.25mM的待测化合物、右美沙芬，混匀。
取398μL的小鼠肝微粒体稀释液加入96孔孵育板中(N＝2)，分别加入2μL 0.25mM的待测化合物、右美沙芬，混匀。
每孔加入300μL预冷的终止液至96孔深孔板中，置于冰上，作为终止板。
将96孔孵育板和NADPH再生系统置于37℃水浴箱中，100转/分钟震荡，预孵5min。从孵育板每孔取出80μL孵育液加入终止板，混匀，补充20μL NADPH再生系统溶液，作为0min样品。再向孵育板每孔加入80μL的NADPH再生系统溶液，启动反应，开始计时。待测化合物反应浓度为1μM，蛋白浓度为0.5mg/mL。
分别于反应10、30、90min时，各取100μL反应液，加入终止板中，涡旋3min终止反应。
将终止板于5000rpm，4℃条件下离心15min。取200μL上清液至预先加入200μL超纯水的96孔板中，混匀，采用LC-MS/MS进行样品分析，进样10uL。
③样品分析方法
本实验采用LC-MS/MS系统检测待测化合物、右美沙芬、奥美拉唑及内标的峰面积，计算化合物与内标峰面积比值。
④数据处理
样品以及内标的峰面积由质谱仪和Analyst软件获得，利用Graphpad prism7.0软件单指数式降解模型对化合物剩余量(R％)与时间作图可得底物消除速率常数K
C t/C 0＝exp(-K*t)
并根据以下公式计算半衰期T 1/2和内在清除率CL int，其中V/M即等于1/C(蛋白)。
Figure PCTCN2022098557-appb-000022
T 1/2(min)；CL int(μL/min/mg)。
实验结果：对本发明化合物及其非氘代的化合物同时测验比较，评价其在人和/或小鼠肝微粒体的代谢稳定性。与非氘代的化合物Binimetinib相比，本发明化合物具有更长的半衰期T 1/2和更低的清除率CL int，可以明显改善代谢稳定性。表性性实施例化合物的结果归纳于表1和2中。
表1：
Figure PCTCN2022098557-appb-000023
表2：
Figure PCTCN2022098557-appb-000024
(3)大鼠药代动力学实验
6只雄性Sprague-Dawley大鼠，7-8周龄，体重约210g，分成2组，每组3只，经静脉或口服单个剂量的化合物(口服10mg/kg)，比较其药代动力学差异。
大鼠采用标准饲料饲养，给予水。试验前16小时开始禁食。药物用PEG400和二甲亚砜溶解。眼眶采血，采血的时间点为给药后0.083小时，0.25小时、0.5小时、1小时、2小时、4小时、6小时、8小时、12小时和24小时。
大鼠吸入乙醚后短暂麻醉，眼眶采集300μL血样于试管。试管内有30μL 1％肝素盐溶液。使用前，试管于60℃烘干过夜。在最后一个时间点血样采集完成之后，大鼠乙醚麻醉后处死。
血样采集后，立即温和地颠倒试管至少5次，保证混合充分后放置于冰上。血样在4℃5000rpm离心5分钟，将血浆与红细胞分离。用移液器吸出100μL血浆到干净的塑料离心管中，标明化合物的名称和时间点。血浆在进行分析前保存在-80℃。用LC-MS/MS测定血浆中本发明化合物的浓度。药代动力学参数基于每只动物在不同时间点的血药浓度进计算。
实验表明，与非氘代的化合物Binimetinib相比，本发明化合物在动物体内具有更好的药代动力学性质，因此具有更好的药效学和治疗效果。
以上内容是结合具体的优选实施方式对本发明所作的进一步详细说明，不能认定本发明的具体实施只局限于这些说明。对于本发明所属技术领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干简单推演或替换，都应当视为属于本发明的保护范围。

Claims (11)

  1. 式(I)化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物：
    Figure PCTCN2022098557-appb-100001
    其中，
    Y 1、Y 2、Y 3、Y 4和Y 5各自独立地选自氢、氘或卤素；
    R 1、R 2、R 3和R 4各自独立地选自氢或氘；
    X各自独立地选自CH 3、CD 3、CHD 2或CH 2D；
    附加条件是，上述化合物至少含有一个氘原子。
  2. 根据权利要求1所述的化合物，其为式(II)化合物：
    Figure PCTCN2022098557-appb-100002
    其中，Y 1、R 1、R 2、R 3、R 4和X如权利要求1所定义；
    或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物。
  3. 根据权利要求1或2所述的化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，Y 1是氘。
  4. 根据权利要求1-3中任一项所述的化合物，或其互变异构体、立体异构体、前药、 晶型、药学上可接受的盐、水合物或溶剂化合物，其中，R 1和R 2是氘。
  5. 根据权利要求1-4中任一项所述的化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，R 3和R 4是氘。
  6. 根据权利要求1-5中任一项所述的化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，其中，X是CD 3。
  7. 根据权利要求1所述的化合物，其选自下式化合物：
    Figure PCTCN2022098557-appb-100003
    Figure PCTCN2022098557-appb-100004
    Figure PCTCN2022098557-appb-100005
    Figure PCTCN2022098557-appb-100006
    Figure PCTCN2022098557-appb-100007
    或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物。
  8. 一种药物组合物，其含有药学上可接受的赋形剂和权利要求1-7中任一项所述的化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物。
  9. 根据权利要求8所述的药物组合物，其还含有另外的治疗剂。
  10. 根据权利要求9所述的药物组合物，其中，另外的治疗剂选自BRAF抑制剂，EGFR抑制剂，EGFR抗体，免疫检查点抑制剂或CDK4/6抑制剂中的一种或几种；
    优选地，BRAF抑制剂选自维莫非尼(vemurafenib)，达拉非尼(dabrafenib)，康奈非尼(encorafenib)，(S)-甲基-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，(S)-(甲基-d 3)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-异丙基-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，(S)-(甲基-d 3)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基)氨基甲酸酯，(S)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基)-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d 5)氨基甲酸甲酯，(S)-(1-((4-(3-(5-氯-2-氟-3-(甲基磺酰氨基)苯基)-1-(丙-2-基-d 7))-1H-吡唑-4-基)嘧啶-2-基)氨基)丙-2-基-1,1,3,3,3-d5)氨基甲酸甲酯；
    优选地，EGFR抑制剂选自吉非替尼(gefitinib)，埃罗替尼(erlotinib)，阿法替尼(afatinib)，达克替尼(dacomitinib)，拉帕替尼(lapatinib)，奥希替尼(osimertinib)，阿美替尼，伏美替尼，CO-1686，WZ4002，PD153035，PF00299804；
    优选地，EGFR抗体选自西妥昔单抗(cetuximab)，帕尼单抗(panitumumab)，耐昔妥珠单抗((Necitumumab)；
    优选地，免疫检查点抑制剂选自帕博利珠单抗(pembrolizumab)，易普利姆妈(ipilimumab)和納武单抗(nivolumab)，阿特珠单抗(atezolizumab)，阿维鲁单抗(avelumab)，德瓦鲁单抗(durvalumab)，吡地利单抗(pidilzumab)；
    优选地，CDK4/6抑制剂选自帕博西尼(palbociclib)，瑞博西尼(ribociclib)，阿贝西尼(abemaciclib)。
  11. 权利要求1-7中任一项所述的化合物，或其互变异构体、立体异构体、前药、晶型、药学上可接受的盐、水合物或溶剂化合物，或权利要求8-10中任一项所述的药物组合物在制备用于治疗MEK激酶介导的疾病的药物中的用途；
    优选地，MEK激酶介导的疾病选自黑色素瘤、急性髓系白血病、神经胶质瘤、神经纤维瘤、非小细胞肺癌、乳腺癌、浆液性癌、胃肠道间质瘤、肺非鳞癌、结直肠癌、胆道癌、骨髓瘤；
    优选地，黑色素瘤选自BRAF V600突变的黑素瘤；
    优选地，结直肠癌选自BRAF V600突变结直肠癌；
    优选地，神经纤维瘤选自神经纤维瘤病1型或丛状神经纤维瘤；
    优选地，BRAF V600突变选自BRAF V600E或BRAF V600K突变。
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1652776A (zh) * 2002-03-13 2005-08-10 阵列生物制药公司 作为mek抑制剂的n3烷基化苯并咪唑衍生物
CN1874768A (zh) * 2003-08-29 2006-12-06 阵列生物制药公司 作为mek抑制剂的n3烷基化苯并咪唑衍生物

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104447701B (zh) * 2013-09-17 2019-03-22 广东东阳光药业有限公司 吡唑类衍生物及其用途
BR112016011222A2 (pt) * 2013-12-12 2017-09-19 Novartis Ag Combinações de trametinib, panitumumab e dabrafenib para o tratamento de câncer
CN111620821B (zh) * 2019-02-27 2022-02-11 深圳市塔吉瑞生物医药有限公司 取代的吡唑类化合物及包含该化合物的组合物及其用途

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1652776A (zh) * 2002-03-13 2005-08-10 阵列生物制药公司 作为mek抑制剂的n3烷基化苯并咪唑衍生物
CN1874768A (zh) * 2003-08-29 2006-12-06 阵列生物制药公司 作为mek抑制剂的n3烷基化苯并咪唑衍生物

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
JIANG WEN-FENG , LI WEN-BAO: "Application of Deuteration in Drug Research", OILU PHARMACEUTICAL, vol. 29, no. 11, 1 January 2010 (2010-01-01), pages 682 - 684, XP008173943, ISSN: 1672-7738 *

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