WO2020125513A1 - 作为cdk抑制剂的大环化合物、其制备方法及其在医药上的应用 - Google Patents

作为cdk抑制剂的大环化合物、其制备方法及其在医药上的应用 Download PDF

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WO2020125513A1
WO2020125513A1 PCT/CN2019/124575 CN2019124575W WO2020125513A1 WO 2020125513 A1 WO2020125513 A1 WO 2020125513A1 CN 2019124575 W CN2019124575 W CN 2019124575W WO 2020125513 A1 WO2020125513 A1 WO 2020125513A1
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cancer
group
alkyl
atom
amino
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PCT/CN2019/124575
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English (en)
French (fr)
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邓永奇
孙健
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凯复制药有限公司
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Priority to US17/413,262 priority Critical patent/US20220017520A1/en
Priority to CN201980022306.8A priority patent/CN111989332B/zh
Priority to KR1020217021032A priority patent/KR20210105375A/ko
Priority to JP2021533362A priority patent/JP2022517723A/ja
Priority to EP19897614.4A priority patent/EP3865488A4/en
Publication of WO2020125513A1 publication Critical patent/WO2020125513A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/68—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member
    • C07D211/72—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D211/74—Oxygen atoms
    • C07D211/76—Oxygen atoms attached in position 2 or 6
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed systems contains four or more hetero rings
    • 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
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • 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
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • 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
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D207/18—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member
    • C07D207/22—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D207/24—Oxygen or sulfur atoms
    • C07D207/26—2-Pyrrolidones
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/06—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/22—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains four or more hetero rings

Definitions

  • the present invention relates to the field of medicine, and relates to a new macrocyclic compound represented by general formula (I), a preparation method thereof, a pharmaceutical composition containing the derivative, and a therapeutic agent thereof, especially as a CDK inhibitor Use in hyperproliferative diseases.
  • a new macrocyclic compound represented by general formula (I) a preparation method thereof, a pharmaceutical composition containing the derivative, and a therapeutic agent thereof, especially as a CDK inhibitor Use in hyperproliferative diseases.
  • CDK Cyclin dependent kinase
  • CDK7, CDK8, CDK9, CDK10, CDK11 and so on Another major category of CDK is involved in transcriptional regulation, mainly including CDK7, CDK8, CDK9, CDK10, CDK11 and so on.
  • CDK7, CDK8, CDK9, CDK10, CDK11 and so on the overexpression or over-activation of cyclin, the inhibition of CDKI activity, and the continuous activation of upstream division signals all cause changes in CDK activity.
  • Dysregulation of CDK activity will directly or indirectly cause uncontrolled cell proliferation, genomic instability (increased DNA mutations, chromosomal deletion, etc.) and chromosomal instability (changes in the number of chromosomes), etc., and participate in the development of tumors.
  • CDK activity is necessary for cell division, and CDK activity is often enhanced in tumor cells, CDK has long been considered as a good target for the development of anti-tumor and other proliferative disorders drugs.
  • LY2835219 (also known as Bemaciclib or Abemaciclib) is a kind of breast cancer therapeutic drug developed by Eli Lilly company with the mechanism of inhibiting cell cycle dependent kinases 4 and 6 (CDK4/6). Its structural formula is The first-phase clinical data show that the drug has a good early effect in the treatment of patients with metastatic breast cancer. Especially for those breast cancer patients with hormone receptor positive, the clinical benefit rate can reach 61%, which means that the time of disease control for patients exceeds After 24 weeks, or the tumor size decreased by more than 30%.
  • PD0332991 (also called Ibrance or Palbociclib) is a CDK4/6 inhibitor developed by Pfizer. Its structural formula is On February 3, 2015, the FDA accelerated the approval of the marketing application for PD0332991, combined with letrozole for the treatment of metastatic breast in postmenopausal women positive for estrogen receptor (ER) and negative for human epidermal growth factor receptor 2 (HER2) cancer.
  • ER estrogen receptor
  • HER2 human epidermal growth factor receptor 2
  • CDK inhibitors can also be used to treat cardiovascular disorders such as restenosis and atherosclerosis and other vascular disorders caused by abnormal cell proliferation.
  • Overexpression of cyclin-dependent kinase inhibitor protein inhibits vascular smooth muscle proliferation and intimal hyperplasia after balloon angioplasty.
  • CDK inhibitors can be used to treat diseases caused by a variety of infectious substances, including fungi, protozoan parasites such as Plasmodium falciparum and DNA and RNA viruses.
  • CDK9 is related to the prevention of HIV replication, so the discovery of new CDK biology continues to emerge new therapeutic indications for CDK inhibitors.
  • CDK is very important in neutrophil-mediated inflammation, and CDK inhibitors promote the regression of inflammation in animal models. Therefore, CDK inhibitors including CDK9 inhibitors can be used as anti-inflammatory agents.
  • CDK inhibitors can be used to improve the effects of various autoimmune disorders.
  • Chronic inflammatory disease-rheumatoid arthritis is characterized by synovial tissue hyperplasia; inhibition of synovial tissue proliferation should minimize inflammation and prevent joint destruction.
  • joint swelling and pain were substantially suppressed by treatment with adenovirus expressing the CDK inhibitor protein p16.
  • CDK inhibitors are effective against other disorders of cell proliferation, including psoriasis (which is characterized by excessive proliferation of keratinocytes), glomerulonephritis, chronic inflammation, and lupus.
  • CDK inhibitors include, for example, WO2015101293A1, WO2016015605A1, WO2016194831A1, WO2008079933A2, and so on. Due to the huge market demand, it is still necessary to continue to develop low-toxic and highly effective CDK inhibitors.
  • Macrocycline has been recognized as an important structural category in drug discovery.
  • the macrocyclic compound itself has a smaller number of rotatable bonds than its non-cyclic analogues, which is a beneficial feature of oral bioavailability (Mallinson, J.; Collins, I. Macrocycles in new drug) discovery.Future Med.Chem.2012,4,1409-1438).
  • macrocyclic compounds are more conformationally restricted than their non-cyclic analogues, which may confer higher target binding and selectivity and improve oral bioavailability.
  • mTOR inhibitors temsirolimus
  • Tubulin stabilizer Ixabepilone
  • Hsp90 inhibitor 17-allylamino-geldanamycin Mcdonald E, Workman P, Jones K. Inhibitors of the HSP90molecular chaperone: attacking the master regulator in cancer.Curr.Top.Med .Chem. 6(11), 1091–1107 (2006)].
  • Pan-CDK inhibitor compound M is another example of a synthetic macrocyclic compound proposed as a development candidate [Hirai H, Takahashi-Suziki I, Shimomura T et al. Potent anti-tumor activity of a macrocycle-quinoxalinone class pan-Cdk inhibitor in vitro and in vivo. Invest. New Drugs 29(4), 534–543 (2011)].
  • the object of the present invention is to provide a compound with excellent CDK inhibitory activity.
  • the inventors have repeatedly conducted serious studies to achieve this goal, and found that a novel class of macrocyclic compounds with excellent CDK inhibitory activity are obtained through intramolecular ring formation reaction. Thus, the present invention has been completed.
  • the present invention more particularly relates to compounds of the following general formula (I):
  • a and B are independently selected from CH and N atoms
  • U, V, W, X and Y are each independently selected from CH and N atoms;
  • R 1 is selected from H atom, C 1-4 alkyl, cyano, C 2-4 alkenyl and C 2-4 alkynyl, wherein the C 1-4 alkyl, C 2-4 alkenyl and C 2 -4 alkynyl groups are each independently optionally substituted with one or more groups selected from halogen, amino, nitro, hydroxyl, and cyano;
  • R 2 is selected from an H atom and a C 1-4 alkyl group, wherein the C 1-4 alkyl group is optionally substituted with one or more groups selected from halogen, amino, nitro, hydroxyl, and cyano;
  • R 1 and R 2 together with the atoms to which they are attached form a C 3-6 cycloalkyl or 3-6 membered heterocyclyl
  • the C 3-6 cycloalkyl and 3-6 membered heterocyclyl are each independently Optionally selected from C 1-4 alkyl, halo C 1-4 alkyl, C 1-4 alkoxy, halo C 1-4 alkoxy, halogen, amino, nitro, hydroxyl and cyano Substituted by one or more groups in
  • R 3 and R 6 are each independently selected from H atom and C 1-4 alkyl group, wherein the alkyl group is optionally selected from C 1-4 alkoxy group, halogenated C 1-4 alkoxy group, halogen, Substituted by one or more of amino, nitro, hydroxyl and cyano
  • R 4 and R 5 are each independently selected from H atom, C 1-4 alkyl, halogen, hydroxy and C 1-4 alkoxy, wherein the C 1-4 alkyl and C 1-4 alkoxy are each Independently optionally substituted with one or more groups selected from halogen, amino, nitro, hydroxyl and cyano; and
  • R 7 is selected from H atom and C 1-4 alkyl group, wherein the alkyl group is optionally selected from C 1-4 alkoxy group, halogenated C 1-4 alkoxy group, halogen, amino, nitro, hydroxyl And one or more groups in the cyano group.
  • a and B are each independently an N atom.
  • Q is a bond or methylene group.
  • U is selected from CH and N atoms.
  • V, X, and Y are each independently CH.
  • W is an N atom.
  • R 1 and R 2 are each independently selected from a H atom and a C 1-4 alkyl group, or R 1 and R 2 and their phases The connected atoms together form C 3-6 cycloalkyl.
  • R 3 and R 6 are each independently C 1-4 alkyl, preferably R 3 is methyl, and R 6 is ethyl base.
  • R 4 and R 5 are each independently a halogen, preferably an F atom.
  • R 7 is a H atom.
  • Typical compounds of the present invention include, but are not limited to:
  • a tautomer a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof.
  • the invention also relates to a method for preparing the compound represented by the general formula (I), which comprises:
  • the compound of general formula (IA) undergoes an intramolecular ring-forming reaction in the presence of a condensing agent to form the compound of general formula (I),
  • LG 1 and LG 2 are independently leaving groups
  • A, B, U, V, W, G, J, L, R 1 to R 6 are as defined in the general formula (I).
  • the condensing agent is selected from 2-(7-benzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), 1 -(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide, N,N'-diisopropylcarbodiimide , O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-azobenzotriazole , O-benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate, 2-(7-azobenzotriazole)-N,N,N',N' -Tetramethylurea hexafluorophosphate, benzotriazol
  • LG 1 and LG 2 are each independently selected from H atom, OH, halogen, methanesulfonate, triflate and p-toluenesulfonate; preferably, LG 1 and LG 2 is independently selected from H atom, OH, Cl atom and Br atom; more preferably, LG 1 is H atom and LG 2 is OH atom.
  • G is -NH-
  • LG 1 is an H atom
  • -LG 2 is an OH atom
  • the present invention also relates to a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of a compound represented by general formula (I) or its tautomer, meso, racemate, enantiomer, non- Enantiomers, or mixtures thereof, or pharmaceutically acceptable salts thereof, and pharmaceutically acceptable carriers, diluents, or excipients.
  • the pharmaceutical composition further comprises one or more additional anti-tumor agents, anti-inflammatory agents, immunosuppressive agents and/or immune checkpoint inhibitors.
  • the pharmaceutical composition further comprises one or more of the following: PTK inhibitor, cyclosporin A, CTLA4-Ig, selected from anti-ICAM-3, anti-IL-2 Receptors, anti-CD45RB, anti-CD2, anti-CD3, anti-CD4, anti-CD80, anti-CD86 and monoclonal antibody OKT3 antibodies, CVT-313, active agents that block the interaction between CD40 and gp39 , Fusion protein constructed from CD40 and gp39, NF- ⁇ B function inhibitor, non-steroidal anti-inflammatory agent, steroids, gold compounds, FK506, mycophenolate mofetil, cytotoxic drugs, TNF- ⁇ inhibitors, Anti-TNF antibodies or soluble TNF receptors, TNF ⁇ , TRAIL, HDAC inhibitors, Gleevec and other inhibitors of signal transduction pathways involved in cell proliferation, inhibitors of cellular hypoxic response, rapamycin, leflunomide Special, cyclooxygenase-2 inhibitors, paclitaxel, c
  • the present invention also relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or Use of a pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, in the preparation of a medicament for treating CDK-related disorders.
  • the present invention also relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing it, is used as a medicine.
  • the present invention also relates to a compound represented by the general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, for the treatment of CDK-related disorders.
  • the present invention also relates to a method for treating CDK-related disorders, which includes administering to a desired patient a therapeutically effective amount of a compound represented by general formula (I) or a tautomer thereof, a racemate, a racemate, a Enantiomers, diastereomers or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing them.
  • a compound represented by general formula (I) or a tautomer thereof a racemate, a racemate, a Enantiomers, diastereomers or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing them.
  • the CDK-related disorder is selected from cancer, inflammation, viral infection, cardiac hypertrophy and HIV.
  • the cancer is selected from bladder cancer, head and neck cancer, breast cancer, stomach cancer, ovarian cancer, colon cancer, lung cancer, brain cancer, laryngeal cancer, lymphatic system cancer, hematopoietic system cancer, urogenital tract Cancer, gastrointestinal cancer, ovarian cancer, prostate cancer, gastric cancer, bone cancer, small cell lung cancer, glioma, colorectal cancer and pancreatic cancer;
  • the inflammation is associated with rheumatoid arthritis, lupus, type 1 diabetes, diabetic nephropathy, multiple sclerosis, glomerulonephritis, chronic inflammation, and organ transplant rejection;
  • the viral infection is associated with HIV virus, human papilloma virus, herpes virus, pox virus, Epstein-Barr virus, Sindse virus or adenovirus.
  • the compounds according to the invention can be administered orally, sublingually, parenterally, subcutaneously, intramuscularly, intravenously, transdermally, topically or rectally.
  • the active ingredient may be the same as conventional
  • the pharmaceutical carriers are mixed together and administered to animals or humans in the form of administration units.
  • Suitable administration unit forms include oral forms such as tablets, gel capsules, powders, granules and oral solutions or suspensions, sublingual or oral administration forms, parenteral, subcutaneous, intramuscular, intravenous, nasal Intra or intraocular administration forms and rectal administration forms.
  • the main active ingredients are mixed with pharmaceutical carriers such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic and the like. Tablets may be coated with sucrose or other suitable materials or processed in such a way that they have prolonged or delayed activity and continuously release a predetermined amount of active ingredient.
  • a gel capsule preparation is obtained by mixing the active ingredient with a diluent and by pouring the obtained mixture into soft or hard capsules.
  • Formulations in the form of syrups or tinctures may contain the active ingredients together with sweeteners, preservatives as well as flavoring agents and suitable coloring agents.
  • Powders or granules dispersible in water may contain the active ingredient, which is mixed with a dispersing agent, a wetting agent or a suspending agent, and a flavoring or sweetening agent.
  • Suppositories are used for rectal administration and are prepared with a binder that melts at rectal temperature, for example, cocoa butter or polyethylene glycol.
  • Aqueous suspensions, isotonic saline solutions or sterile and injectable solutions which contain pharmacologically compatible dispersants and/or wetting agents are used parenterally, intranasally or intraocularly Apply.
  • the active ingredient (possibly with one or more additive carriers) can also be formulated as a microcapsule.
  • the compounds of the present invention can be used in doses between 0.01 mg/day and 1000 mg/day, provided in a single dose/day or administered in several doses throughout the day, for example, the same dose twice a day .
  • the daily dose administered is advantageously between 0.1 mg and 100 mg, even more advantageously between 2.5 mg and 50 mg. It may be necessary to use dosages outside these ranges, as those skilled in the art will recognize this.
  • the pharmaceutical composition may also be formulated for external administration. It can be introduced into the common forms of this type of application (ie, in particular lotions, foams, gels, dispersants, sprays), said common forms having excipients, said excipients in particular It can penetrate the skin in order to improve the properties and accessibility of the active ingredients.
  • these compositions generally further comprise a physiologically acceptable medium, which usually contains water or a solvent, for example, alcohol, ether or ethylene glycol.
  • the composition may also contain surfactants, preservatives, stabilizers, emulsifiers, thickeners, other active ingredients that produce complementary effects or possible synergistic effects, trace elements, essential oils, fragrances, colorants, collagen, Chemical or mineral filters.
  • stereoisomer refers to geometric isomer (or configurational isomer) or optical isomer.
  • Geometric isomers are caused by substituents at different positions on the double bond, which can then have a Z or E configuration, also known as cis or trans.
  • optical isomers are caused in particular by substituents at different spatial positions on the carbon atom, which contains four different substituents. This carbon atom constitutes the chiral center or asymmetric center.
  • Optical isomers include diastereomers and enantiomers. Optical isomers that are non-overlapping mirror images of each other are called “enantiomers”. Optical isomers that are not mirror images that can be overlapped with each other are called “diastereomers”.
  • racemic mixture A mixture containing two separate enantiomeric forms of equal amounts and opposite chirality is called a "racemic mixture”.
  • tautomer refers to a structural isomer of a compound obtained by prototropie, that is, migration of a hydrogen atom and change in the position of a double bond.
  • the different tautomers of the compound are generally interconvertible and are present in a balanced solution in proportions, which may vary depending on the solvent used, temperature or pH.
  • pharmaceutically acceptable is understood to mean that it is used to prepare a pharmaceutical composition, the composition is generally safe, non-toxic, meets the needs in biology or other aspects and the combination The substance can be accepted for veterinary and human drug use.
  • a "pharmaceutically acceptable salt” of a compound is understood to refer to the following salts, which are pharmaceutically acceptable (as defined herein) salts and which possess the expected pharmacological activity of the parent compound.
  • This salt includes:
  • Acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., or organic acids such as acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid , Fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, hydroxynaphthoic acid, 2-hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucon Acid addition salts formed by acids, 2-naphthalenesulfonic acid, propionic acid, salicylic acid, succinic acid, dibenzoyl-L-tartaric acid, tartaric acid, p-toluenesulfonic acid, trimethylacetic acid, trifluoroacetic acid, etc.; with
  • alkali metal ions for example, Na + , K + or Li +
  • alkaline earth metal ions for example Ca 2+ or Mg 2+
  • aluminum ions or salts formed when coordinated with organic or inorganic bases.
  • Acceptable organic bases include diethanolamine, ethanolamine, N-methylglucamine, triethanolamine, tromethamine, and the like.
  • Acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, and sodium hydroxide.
  • halogen refers to a fluorine, bromine, chlorine or iodine atom.
  • C 1-4 alkyl refers to a saturated linear or branched hydrocarbon chain containing 1 to 4 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl groups.
  • C 1-4 alkylene refers to a divalent hydrocarbon chain containing 1 to 4 carbon atoms. Representative examples include, but are not limited to, CH 2 -, -CH 2 -CH 2 -, -CH 2 -CH 2 -CH 2 -, -CH 2 -CH 2 -CH 2 -CH 2 -, and the like.
  • C 2 -4 alkenyl refers to a straight or branched hydrocarbon chain having at least one double bond and having 2 to 4 carbon atoms. Representative examples include, but are not limited to, vinyl, propenyl, butenyl, and the like.
  • C 2 -4 alkynyl refers to a straight or branched hydrocarbon chain having at least one triple bond and having 2 to 4 carbon atoms. Representative examples include, but are not limited to, ethynyl, propynyl, butynyl, and the like.
  • C 1-4 alkoxy refers to -O-(C 1-4 alkyl), wherein C 1-4 alkyl is as defined above.
  • Non-limiting examples include methoxy, ethoxy, propoxy, butoxy, and the like.
  • halogenated C 1-4 alkyl means that C 1-4 alkyl is substituted with one or more halogens, wherein C 1-4 alkyl and halogen are as defined above.
  • halo C 1-4 alkoxy means that C 1-4 alkoxy is substituted with one or more halogen, wherein C 1-4 alkoxy and halogen are as defined above.
  • C 3-6 cycloalkyl refers to a saturated or partially unsaturated monocyclic hydrocarbon system containing 3 to 6 carbon atoms. Representative examples include, but are not limited to, cyclohexyl, cyclopentyl, cyclobutyl, Cyclopropyl, cyclohexenyl, etc.
  • 3-6 membered heteroalkyl refers to those containing 3 to 6 ring atoms, of which 1-3 ring atoms are selected from nitrogen, oxygen or S(O) m (where m is 0, 1 or 2)
  • Heteroatom saturated or partially unsaturated monocyclic hydrocarbon systems representative examples include, but are not limited to, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, dihydroimidazolyl, Dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, etc.
  • hydroxyl refers to the -OH group.
  • nitro refers to -NO 2.
  • amino refers to -NH 2.
  • cyano refers to -CN.
  • a bond refers to a covalent bond represented by "-”.
  • the term "leaving group” refers to a chemical group that can be easily replaced by a nucleophilic reagent during a nucleophilic substitution reaction, for example, an H atom, an OH, a halogen atom such as a chlorine atom or a bromine atom, or a sulfonate.
  • the sulfonate can be particularly mesylate (-OS(O 2 )-CH 3 ), trifluoromethanesulfonate (-OS(O) 2 -CF 3 ) or p-toluenesulfonate (-OS( O) 2 -(p-Me-C 6 H 4 )).
  • heterocyclic group optionally substituted with an alkyl group means that an alkyl group may but need not be present, and the description includes the case where the heterocyclic group is substituted with an alkyl group and the case where the heterocyclic group is not substituted with an alkyl group .
  • Substituted refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3 hydrogen atoms are independently substituted with a corresponding number of substituents. It goes without saying that the substituents are only at their possible chemical positions, and those skilled in the art can determine (through experiment or theory) possible or impossible substitutions without undue effort. For example, an amino group or hydroxyl group having free hydrogen may be unstable when combined with a carbon atom having an unsaturated (eg, olefinic) bond.
  • “Pharmaceutical composition” means a mixture containing one or more compounds described herein or a physiological/pharmaceutically acceptable salt or prodrug thereof with other chemical components, and other components such as physiological/pharmaceutically acceptable carriers And excipients.
  • the purpose of the pharmaceutical composition is to promote the administration to the living body, facilitate the absorption of the active ingredient and thereby exert the biological activity.
  • the structure of the compound is determined by nuclear magnetic resonance (NMR) or/and mass spectrometry (MS).
  • NMR chemical shift ( ⁇ ) is given in units of 10 -6 (ppm).
  • the measurement solvent was deuterated chloroform (CDCl 3 ), and the internal standard was tetramethylsilane (TMS).
  • TMS tetramethylsilane
  • s is a singlet
  • bs is a broad singlet
  • d is a doublet
  • t is a triplet
  • qdt is a quartet
  • m is a multiplet or a large number of peaks
  • dd is a double doublet, etc.
  • LC/MS Agilent LCMS1260/MSD6120, column: Agilent ZORBAX SB-C18, 2.1*50mm, 1.8 ⁇ m, mobile phase: A: H 2 O (0.1% FA), B: acetonitrile, gradient elution, 0.5 mL/min, 45.0°C, ionization mode: API-ES, polarity: positive.
  • NMR instrument Bruker ARX-500 and Bruker ARX-400.
  • MTT testing instrument Thermo Scientific Multiskan GO full wavelength microplate reader.
  • the thin-layer chromatography silica gel plate uses Qingdao GF254 silica gel plate, the thin-film chromatography (TLC) silica gel plate uses a specification of 0.15mm ⁇ 0.2mm, and the thin-layer chromatography separation purification product uses a specification of 0.4mm ⁇ 0.5mm.
  • the reactions are carried out under an argon atmosphere or a nitrogen atmosphere.
  • the solution in the reaction refers to an aqueous solution.
  • reaction temperature is room temperature.
  • the reaction raw material 3-aminobutyric acid 107 in the sixth step was replaced with 3-amino-3-methylbutyric acid to obtain 4 5 -((4-ethylpiperazine-1 -Yl)methyl)-1 4 ,2 5 -difluoro-1 2 ,8,8-trimethyl- 1 1 H-3,5-diaza-1(6,1)-benzo(d ]Imidazole-2(4,2)-pyrimidine-4(2,4)-pyridinecyclooctan-6-one 8, white solid.
  • the reaction raw material 3-aminobutyric acid 107 in the sixth step was replaced with 2-(1-aminocyclopropyl)acetic acid to obtain 5'-((4-ethylpiperazine- 1-yl)methyl)-4',5'-difluoro-2'-methylspiro[cyclopropane-1,8'-3,5-diaza-1(6,1)-benzo[ d]Imidazole-2(4,2)-pyrimidine-4(2,4)-pyridinecyclooctan]-6'-one 9, white solid.
  • the IC 50 measurement method for CDK4/Cyclin D1 is carried out as follows.
  • a 96-well filter plate (Millipore MADVN6550) was used. The total volume is 0.1ml, containing buffer A (20mM TRIS (tris[hydroxymethyl]aminomethane) (pH7.4), 50mM NaCl, 1mM dithiothreitol, 10mM MgCl2), 25 ⁇ M ATP (containing 0.25 ⁇ Ci[ 32P] ATP), 20 ng CDK4, 1 ⁇ g of retinoblastoma protein and test compound in the appropriate ratio of buffer A dilution. As a control with no inhibitory effect, buffer A alone with no test compound added was used.
  • Buffer A containing excess EDTA was used to determine the background 32P level in the absence of enzyme activity.
  • TCA trichloroacetic acid
  • the plate was kept at 4°C for at least 1 hour to allow the substrate to precipitate.
  • the wells were then washed 5 times with 0.2 ml of 10% TCA, and 32P binding was measured using a beta plate counter (wallac Inc., Gaithersburg, MD).
  • the IC 50 of the test compounds was determined by the median effect method (Chou, TC and Talalay P. Applications of the median effect principle for the assessment of low-dose risk of carcinogens and for the quantitation of synergism and antagonism of chemotherapeutic agents. In New Avenues in Developmental Cancer Chemotherapy (Eds. Harrap, KTand Connors, TA), pp. 37-64. Academic Press, New York, 1987).
  • HEPES HEPES with a concentration of 1000 mM and a pH of 7.5 and 50 ⁇ L of Brij-35 with a concentration of 30% were added to 949.95 mL of ultrapure water and mixed.
  • coating solution Reagent#3 (included in the 12-sipper chip used by Caliper instrument), 50 mL of HEPES with a concentration of 1000 mM and pH of 7.5, and 50 mL of the mother solution with a concentration of 4%.
  • 0.5M EDTA and 0.25mL Brij-35 with a mother liquor concentration of 30% were added to 374.75mL ultrapure water and mixed well.
  • CDK4/D3 enzyme solution Take 7 ⁇ L of CDK4/D3 enzyme solution and 9 ⁇ L of 1M DTT in 1M mother solution, add them to 1784 ⁇ L of 1x CDK4 kinase buffer, and mix well.
  • a 10 mM DMSO stock solution of the test article was taken and diluted with DMSO to prepare a solution with a concentration of 50 ⁇ M as the mother liquor.
  • the above mother liquor was diluted four-fold with DMSO to make solutions of 12.5 ⁇ M, 3.125 ⁇ M, 0.78 ⁇ M, 0.195 ⁇ M, 0.0488 ⁇ M, 12.2nM, 3nM, 0.76nM, 0.19nM, and then each concentration was buffered by 1x kinase
  • the solution was diluted 10-fold to make a 5-fold compound solution.
  • the Caliper instrument reads the data, calculates the inhibition rate by the following formula, and then uses GraphPad5.0 software for curve fitting to obtain the IC 50 value.
  • BrdU method (BrdU cell proliferation test kit, Cell Signaling Technology company) was used to detect cell proliferation
  • the 20-fold concentration of the mother liquor was diluted with ultrapure water to one-fold.
  • the BrdU detection antibody with a mother liquor concentration of 100-fold is diluted with a detection antibody diluent to a 1-fold detection antibody solution.
  • the anti-mouse IgG and HRP-labeled antibody with a mother liquor concentration of 100-fold were diluted with HRP-labeled antibody diluent to a 1-fold HRP-labeled secondary antibody solution.
  • the BrdU solution with a mother liquor concentration of 1,000 times is diluted with the medium corresponding to the cells to a 10-fold BrdU solution.
  • test compound mother liquor formulated with 100% DMSO into 10 mM mother liquor.
  • test compound gradient dilution solution 10 mM test compound mother solution was serially diluted 4 times with DMSO at concentrations of 2.5 mM, 625 ⁇ M, 156 ⁇ M, 39 ⁇ M, 9.8 ⁇ M, 2.5 ⁇ M. 2 ⁇ L of DMSO diluted compounds were added to 198 ⁇ L of culture medium containing 10% FBS to prepare a 10-fold test substance. The highest concentration of the test substance was 100 ⁇ M, and the DMSO concentration was 1%, for a total of 7 concentration gradients.
  • MCF-7 medium DMEM + 10% FBS + 0.01 mg/mL insulin
  • MDA-MB-435S and U87MG cells were added with FBS (9 ⁇ L) per well, and MCF-7 cells were added with 8 ⁇ L FBS per well to make the final concentration of FBS 10%;
  • test substance (10 ⁇ L) with different concentrations to each well, so that the final concentration of the test substance is 10 ⁇ M, 2.5 ⁇ M, 625 nM, 156 nM, 39 nM, 9.8 nM, 2.5 nM, 3 replicate wells/group, and culture at 37°C 72 hours;
  • Normal cell control normal cells without any treatment
  • OD blank control blank control value
  • OD normal cell control normal cell control value

Abstract

涉及作为CDK抑制剂的大环化合物、其制备方法及其在医药上的应用。具体而言，涉及一种通式(I)所示的新的大环化合物、其制备方法及含有该化合物的药物组合物以及其作为治疗剂，特别是作为CDK抑制剂的用途以及用于治疗癌症、炎症、病毒感染、心脏肥大或HIV的用途，其中通式（I）的各取代基与说明书中的定义相同。

Description

作为CDK抑制剂的大环化合物、其制备方法及其在医药上的应用 技术领域
本发明涉及属于医药领域，涉及一种通式(I)所示的新的大环化合物、其制备方法及含有该衍生物的药物组合物以及其作为治疗剂，特别是作为CDK抑制剂在治疗过度增殖性疾病中的用途。
背景技术
近年来，肿瘤已超越心血管疾病，成为全球第一大死亡疾病，抗肿瘤药物研究具有重要的学术和现实意义。过度活化、持续的细胞增殖是肿瘤的一个基本特征，因此诱导细胞周期阻滞可有效抑制肿瘤的生长。细胞周期蛋白依赖性激酶(cyclin dependent kinase，CDK)属于丝/苏氨酸蛋白激酶家族，是参与细胞周期调节的关键激酶。目前已报道有20个不同的CDK，根据CDK功能的不同，可以将其主要分为两大类。一类CDK参与细胞周期调控，主要包括CDK1、CDK2、CDK4、CDK6等。另一大类CDK参与转录调节，主要包括CDK7、CDK8、CDK9、CDK10、CDK11等。在肿瘤细胞中，细胞周期蛋白(cyclin)过表达或过度活化、CDKI活性被抑制、上游***信号持续激活等都会引起CDK的活性改变。CDK活性失调会直接或间接引起细胞增殖失控、基因组不稳定(DNA突变增加，染色体缺失等)和染色体不稳定(染色体数目变化)等，参与肿瘤的发生发展。由于CDK活性为细胞***所必需，而在肿瘤细胞中又常有CDK活性增强，因此长期以来，CDK一直被认为是抗肿瘤及其他增殖失调疾病药物研发的较好靶点。
LY2835219(也称作Bemaciclib或Abemaciclib)是由礼来(Eli Lilly)公司开发的一种以抑制细胞周期依赖性激酶4和6(CDK4/6)为机理的乳腺癌治疗药物。其结构式为
Figure PCTCN2019124575-appb-000001
一期临床数据表明该药单药治疗转移性乳腺癌患者早期疗效较好，尤其对于那些激素受体阳性的乳腺癌患者，临床获益率可达61％，这意味着患者疾病控制的时间超过了24周，或肿瘤大小下降30％以上。目前,美国FDA已经批准礼来(Eli Lilly)的Abemaciclib(VERZENIO)联合非甾体芳香酶抑制剂(non-steroidal aromatase iinhibitors,NSAI)来曲唑或阿那曲唑用于一线治疗绝经后激素受体(HR)阳性，人表皮生长因子受体2(HER2)阴性的晚期或转移性乳腺癌。非小细胞肺癌的临床研究也已开始。
PD0332991(也称作Ibrance或Palbociclib)是由辉瑞(Pfizer)公司开发的 CDK4/6抑制剂。其结构式为
Figure PCTCN2019124575-appb-000002
2015年2月3日，FDA加速批准了PD0332991的上市申请，与来曲唑联用治疗***受体(ER)阳性、人表皮生长因子受体2(HER2)阴性的绝经后妇女转移性乳腺癌。
CDK抑制剂也可用于治疗心血管病症如再狭窄和动脉粥样硬化和由异常细胞增殖所致的其他血管病症。细胞周期蛋白依赖性激酶抑制剂蛋白的过表达抑制气囊血管成形术之后的血管平滑肌增殖和内膜增生。而且，嘌呤CDK2抑制剂CVT-313(Ki＝95nM)使得大鼠新内膜生成的抑制大于80％。
CDK抑制剂可用于治疗由多种感染性物质引起的疾病，包括真菌、原生动物寄生虫如恶性疟原虫和DNA与RNA病毒。
最近，CDK9与HIV复制的预防有关，因此新CDK生物学的发现继续涌现出CDK抑制剂的新治疗适应症。
CDK在嗜中性粒细胞介导的炎症中非常重要，且CDK抑制剂促进动物模型中炎症的消退。因此，包括CDK9抑制剂的CDK抑制剂可用作抗炎剂。
选择性CDK抑制剂可用于改善各种自身免疫性病症的影响。。慢性炎性疾病-类风湿性关节炎的特征为滑液组织增生；滑液组织增殖的抑制应将炎症降到最小并预防关节破坏。在大鼠关节炎模型中，通过用表达CDK抑制剂蛋白p16的腺病毒治疗而基本上抑制了关节肿痛。CDK抑制剂对抗细胞增殖的其他病症是有效的，包括银屑病(其特征为角质化细胞过度增殖)、肾小球肾炎、慢性炎症和狼疮。
目前已有的CDK抑制剂专利申请包括，如WO2015101293A1、WO2016015605A1、WO2016194831A1、WO2008079933A2等等。由于巨大的市场需求，所以仍有必要继续研发低毒、高效的CDK抑制剂。
大环素已被公认为药物发现中的重要结构类别。对于相同数量的重原子，大环化合物本身具有比其非环状类似物更少数量的可旋转键，这是口服生物利用度的有益特征(Mallinson,J.；Collins,I.Macrocycles in new drug discovery.Future Med.Chem.2012,4,1409-1438)。结果，大环化合物比其非环状类似物更具构象限制，这可能赋予更高的靶标结合和选择性并改善口服生物利用度。
天然产物大环化合物的开发已经产生了几种肿瘤药物，这些药物被批准用于临床或已经达到后期临床开发，例如mTOR抑制剂
Figure PCTCN2019124575-appb-000003
(temsirolimus)[Kwitkowski VE,Prowell TM,Ibrahim A et al.FDA approval summary:temsirolimus as treatment for advancedrenal cell carcinoma.Oncologist 15(4),428–435(2010)]，微管蛋白稳定剂
Figure PCTCN2019124575-appb-000004
(伊沙匹隆)和Hsp90抑制剂17-烯丙基氨基-格尔德霉素[Mcdonald E,Workman P,Jones K.Inhibitors of the HSP90molecular chaperone: attacking the master regulator in cancer.Curr.Top.Med.Chem.6(11),1091–1107(2006)]。许多合成大环化合物已进入临床开发阶段，例如双重JAK2/FLT3抑制剂pacritinib，目前正在进行II期临床试验[Hart S,Goh KC,Novotny-Diermayr V et al.SB1518,a novel macrocyclic pyrimidinebased JAK2inhibitor for the treatment of myeloid and lymphoid malignancies.Leukemia 25(11),1751–1759(2011)]，II期临床试验中的CDK2/JAK2/FLT3抑制剂SB1317[William AD,Lee ACH,Goh KC et al.Discovery of kinase spectrum selective macrocycle(16E)-14-methyl-20oxa5,7,14,26-tetraazatetracyclo[19.3.1.1(2,6).1(8,12)]heptacosa1(25),2(26),3,5,8(27),9,11,16,21,23-decaene(SB1317/TG02),a potent inhibitor of cyclin dependent kinases(CDKs),janus kinase 2(JAK2),and Fms-like tyrosine kinase-3(FLT3)for the treatment of cancer.J.Med.Chem.55(1),169–196(2012)]和西仑吉肽(cilengitide)，一种用于治疗胶质母细胞瘤的III期临床试验中的合成环肽[Stupp R,Van Den Bent MJ,Erridge SC et al.Cilengitide in newly diagnosed glioblastoma with MGMT promoter methylation:Protocol of a multicenter,randomized,open-label,controlled phase III trial(CENTRIC).J.Clin.Oncol.28(15s),TPS152(2010)]。泛CDK抑制剂化合物M是作为开发候选物提出的合成大环化合物的另一个实例[Hirai H,Takahashi-Suziki I,Shimomura T et al.Potent anti-tumor activity of a macrocycle-quinoxalinone class pan-Cdk inhibitor in vitro and in vivo.Invest.New Drugs 29(4),534–543(2011)]。
发明内容
本发明的目的在于提供具有优异CDK抑制活性的化合物，发明人为了达到该目的反复进行了认真研究，结果发现经过分子内成环反应获得了一类新颖的具有优异CDK抑制活性的大环化合物，从而完成了本发明。
本发明更特别地涉及下面通式(I)的化合物：
Figure PCTCN2019124575-appb-000005
或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐，
其中：
A和B各自独立地选自CH和N原子；
U、V、W、X和Y各自独立地选自CH和N原子；
Q选自一个键、C 1-4亚烷基和-C(＝O)-，所述C 1-4亚烷基任选被选自C 1-4烷基、卤代C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
G选自一个键、-O-、-N(R 7)-、-C(＝O)-、C 1-4亚烷基、C 2-4烯基、-S-、-SO-、-SO 2-和3-6元杂环基，其中所述C 1-4亚烷基、C 2-4烯基和3-6元杂环基各自独立地任选被选自C 1-4烷基、卤代C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
J选自一个键、-O-、-N(R 7)-、-C(＝O)-、C 1-4亚烷基、C 2-4烯基、-S-、-SO-、-SO 2-和3-6元杂环基，其中所述C 1-4亚烷基、C 2-4烯基和3-6元杂环基各自独立地任选被选自C 1-4烷基、卤代C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
L选自一个键、-C(＝O)-、C 1-4亚烷基、C 2-4烯基、-S-、-SO-、-SO 2-和3-6元杂环基，其中所述C 1-4亚烷基、C 2-4烯基和3-6元杂环基各自独立地任选被选自C 1-4烷基、卤代C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
R 1选自H原子、C 1-4烷基、氰基、C 2-4烯基和C 2-4炔基，其中所述C 1-4烷基、C 2-4烯基和C 2-4炔基各自独立地任选被选自卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
R 2选自H原子和C 1-4烷基，其中所述C 1-4烷基任选被选自卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
或者，R 1和R 2与其相连接的原子一起形成C 3-6环烷基或3-6元杂环基，所述C 3-6环烷基和3-6元杂环基各自独立地任选被选自C 1-4烷基、卤代C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
R 3和R 6各自独立地选自H原子和C 1-4烷基，其中所述烷基任选被选自C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代
R 4和R 5各自独立地选自H原子、C 1-4烷基、卤素、羟基和C 1-4烷氧基，其中所述C 1-4烷基和C 1-4烷氧基各自独立地任选被选自卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；和
R 7选自H原子和C 1-4烷基，其中所述烷基任选被选自C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代。
在一个优选实施例方案中，在通式(I)所示的化合物中，-G-J-L-选自-NH-CH 2-CH 2-、-O-CH 2-CH 2-、-NH-C(＝O)-CH 2-和-C(＝O)-NH-CH 2-。
在另一个优选实施例方案中，在通式(I)所示的化合物中，A和B各自独立地为N原子。
在另一个优选实施例方案中，在通式(I)所示的化合物中，Q为一个键或亚甲基。
在另一个优选实施例方案中，在通式(I)所示的化合物中，U选自CH和N原子。
在另一个优选实施例方案中，在通式(I)所示的化合物中，V、X和Y各自独立地为CH。
在另一个优选实施例方案中，在通式(I)所示的化合物中，W为N原子。
在另一个优选实施例方案中，在通式(I)所示的化合物中，R 1和R 2各自独立地选自H原子和C 1-4烷基，或者，R 1和R 2与其相连接的原子一起形成C 3-6环烷基。
在另一个优选实施例方案中，在通式(I)所示的化合物中，R 3和R 6各自独立地为C 1-4烷基，优选地R 3为甲基，且R 6为乙基。
在另一个优选实施例方案中，在通式(I)所示的化合物中，R 4和R 5各自独立地为卤素，优选F原子。
在另一个优选实施例方案中，在通式(I)所示的化合物中，R 7为H原子。
本发明的典型化合物包括但不限于：
Figure PCTCN2019124575-appb-000006
Figure PCTCN2019124575-appb-000007
Figure PCTCN2019124575-appb-000008
或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐。
本发明还涉及一种制备通式(I)所示的化合物的方法，其包括：
Figure PCTCN2019124575-appb-000009
通式(IA)化合物在缩合剂存在下发生分子内成环反应形成通式(I)化合物，
其中：
LG 1和LG 2各自独立地为离去基；
A、B、U、V、W、G、J、L、R 1至R 6如通式(I)中所定义。
在一个优选实施例方案中，所述缩合剂选自2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU)、1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N,N'-二环己基碳化二亚胺、N,N'-二异丙基碳二酰亚胺、O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸酯、1-羟基苯并***、1-羟基-7-偶氮苯并三氮唑、O-苯并三氮唑-N,N,N',N'-四甲脲六氟磷酸酯、2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯、苯并三氮唑-1-基氧基三(二甲基氨基)磷鎓六氟磷酸盐和六氟磷酸苯并***-1-基-氧基三吡咯烷基磷，优选2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU)。
在另一个优选实施例方案中，LG 1和LG 2各自独立地选自H原子、OH、卤素、甲磺酸酯、三氟甲磺酸酯和对甲苯磺酸酯；优选地，LG 1和LG 2各自独立地选自H原子、OH、Cl原子和Br原子；更优选地，LG 1为H原子，LG 2为OH原子。
在另一个优选实施例方案中，G为-NH-，LG 1为H原子，J为-C(＝O)-，且-LG 2为OH原子。
本发明还涉及一种药物组合物，其包含治疗有效量的通式(I)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐，以及药学上可接受的载体、稀释剂或赋形剂。
在一个优选实施例方案中，所述药物组合物还包含一种或多种额外的抗肿瘤剂、抗炎剂、免疫抑制剂和/或免疫检测点抑制剂。
在另一个优选实施例方案中，所述药物组合物还包含以下一种或多种：PTK抑制剂、环孢菌素A、CTLA4-Ig、选自抗-ICAM-3、抗-IL-2受体、抗-CD45RB、抗-CD2、抗-CD3、抗-CD4、抗-CD80、抗-CD86和单克隆抗体OKT3的抗体、CVT-313、阻断CD40与gp39之间相互作用的活性剂、由CD40和gp39构建的融合蛋白、NF-κB功能抑制剂、非甾体抗炎剂、类固醇、金化合物、FK506、麦考酚酸吗啉乙酯、细胞毒性药、TNF-α抑制剂、抗-TNF抗体或可溶性TNF受体、TNFα、TRAIL、HDAC抑制剂、格列卫和其他涉及细胞增殖的信号转导途径的抑制剂、细胞低氧应答抑制剂、雷帕霉素、来氟米特、环氧合酶-2抑制剂、紫杉醇、顺铂、卡铂、阿霉素、去甲柔红霉素、柔红霉素、氨基蝶呤、氨甲蝶呤、甲基叶酸、丝裂霉素C、海鞘素743、泊非霉素、5-氟尿嘧啶、6-巯嘌呤、吉西他滨、阿糖胞苷、鬼臼毒素、依托泊苷、磷酸依托泊苷、替尼泊苷、美法仑、长春碱、长春新碱、异长春碱、埃坡霉素、长春地辛、环氧长春碱、PD-1、PDL-1或其衍生物。
本发明还涉及通式(I)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体或其混合物形式，或其药学上可接受的盐，或包含其的药物组合物在制备治疗CDK相关病症的药物中的用途。
本发明还涉及通式(I)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体或其混合物形式，或其药学上可接受的盐，或包含其的药物组合物，其用作药物。
本发明还涉及通式(I)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体或其混合物形式，或其药学上可接受的盐，或包含其的药物组合物，其用于治疗CDK相关病症。
本发明还涉及一种治疗CDK相关病症的方法，包括给予所需患者治疗有效量的通式(I)所示的化合物或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体或其混合物形式，或其药学上可接受的盐，或包含其的药物组合物。
在一个优选实施例方案中，所述CDK相关病症选自癌症、炎症、病毒感染、心脏肥大和HIV。
在一个更优选实施例方案中，所述癌症选自膀胱癌、头颈癌、乳腺癌、胃癌、卵巢癌、结肠癌、肺癌、脑癌、喉癌、淋巴***癌、造血***癌、泌尿生殖道癌、胃肠癌、卵巢癌、***癌、胃癌、骨癌、小细胞肺癌、神经胶质瘤、结肠直肠癌和胰腺癌；
所述炎症与类风湿性关节炎、狼疮、1型糖尿病、糖尿病性肾病变、多发性硬化、肾小球肾炎、慢性炎症和器官移植排斥有关；
所述病毒感染与HIV病毒、人***状瘤病毒、疱疹病毒、痘病毒、EB病毒、 新培斯病毒或腺病毒相关。
根据本发明的化合物可以被口服施用、舌下施用、肠胃外施用、皮下施用、肌内施用、静脉内施用、经皮施用、局部施用或直肠施用。
在本发明的药用化合物中，对于口服施用、舌下施用、肠胃外施用、皮下施用、肌内施用、静脉内施用、经皮施用、局部施用或直肠施用而言，活性成分可以与常规的药用载体混合在一起，以施用单位的形式施用于动物或人类。适合的施用单位形式包含口服形式如片剂、凝胶胶囊剂、粉剂、颗粒剂和口服的溶液剂或混悬剂，舌下或口腔施用形式，肠胃外、皮下、肌内、静脉内、鼻内或眼内施用形式和直肠施用形式。
当固体组合物被制备成片剂形式时，主要活性成分与药用载体如明胶、淀粉、乳糖、硬脂酸镁、滑石、***胶等混合。片剂可以采用蔗糖或其他适合的材料包衣或者以如此的方式处理以至于其具有延长的或延迟的活性并且连续释放预定量的活性成分。
通过将活性成分与稀释剂混合并通过将获得的混合物倾倒入软质或硬质胶囊中来获得凝胶胶囊制剂。
糖浆剂或酊剂形式的制剂可以包含活性成分连同甜味剂、防腐剂以及芳香剂和适当的着色剂。
可分散于水中的粉剂或颗粒剂可以包含活性成分，其与分散剂、润湿剂或悬浮剂以及与矫味剂或甜味剂混合在一起。
栓剂用于直肠施用，其采用在直肠温度下熔化的粘合剂，例如，可可脂或聚乙二醇来制备。
水性混悬剂、等渗的生理盐水溶液剂或无菌的且可注射的溶液剂(其包含药理学上可兼容的分散剂和/或润湿剂)用于肠胃外、鼻内或眼内施用。
活性成分(可能与一种或多种添加剂载体一起)也可以被配制成微囊剂。
本发明的化合物能够以介于0.01mg/天和1000mg/天之间的剂量来使用，以单一剂量/天的方式来提供或者以全天内若干剂量的方式来施用，例如，相同剂量每天两次。所施用的日剂量有利地介于0.1mg和100mg之间，甚至更有利地介于2.5mg和50mg之间。使用超出这些范围的剂量可能是需要的，本领域技术人员自身将会意识到这一点。
在本发明的一个特定实施方案中，药物组合物也可以被配制用于外部施用。它可以被引入到该施用类型的常用形式(即，特别是洗剂、泡沫剂、凝胶剂、分散剂、喷雾剂)中，所述常用形式具有赋形剂，所述赋形剂特别地能够穿透皮肤，以便于改善活性成分的性质和可接近性。除了根据本发明的组合物之外，这些组合物通常进一步包含生理上可接受的介质，所述介质通常包含水或溶剂，例如，醇、醚或乙二醇。所述组合物还可以包含表面活性剂、防腐剂、稳定剂、乳化剂、增稠剂、产生互补效果或可能的协同效果的其他活性成分、微量元素、精油、香 料、着色剂、胶原蛋白、化学或矿物过滤剂。
定义
除非有相反陈述，否则下列用在说明书和权利要求书中的术语具有下述含义。
在本发明的含义内，“立体异构体”是指几何异构体(或构型异构体)或旋光异构体。
几何异构体由双键上不同位置的取代基所导致，然后其可以具有Z或E构型，也被称作顺式或反式。
光学异构体特别地由碳原子上不同空间位置的取代基所导致，所述碳原子包含四个不同的取代基。这个碳原子则构成手性中心或不对称中心。光学异构体包括非对映异构体和对映异构体。彼此为不可重叠的镜像的光学异构体被称作“对映异构体”。彼此不为可重叠的镜像的光学异构体被称作“非对映异构体”。
含有等量的、相反手性的两种单独的对映异构体形式的混合物被称作“外消旋混合物”。
在本发明的含义内，“互变异构体”是指通过质子转移重排(prototropie)，即通过氢原子的迁移和双键的位置的改变而获得的化合物的结构异构体。化合物的不同互变异构体通常是可互相转化的，并且按比例平衡地存在于溶液中，所述比例可以根据所使用的溶剂、温度或pH而变化。
在本发明中，“药学上可接受的”被理解为是指其用于制备药物组合物，所述组合物一般是安全的，无毒的，在生物学或其他方面满足需要并且所述组合物可以被接受用于兽类和人类药物用途。
在本发明中，化合物的“药学上可接受的盐”被理解为指代下列盐，其是药学上可接受的(如本文所定义的)盐并且其具备预期的母体化合物的药理活性。这种盐包括：
(1)与无机酸如盐酸、氢溴酸、硫酸、硝酸、磷酸等形成的酸加成盐，或与有机酸如乙酸、苯磺酸、苯甲酸、樟脑磺酸、柠檬酸、乙磺酸、富马酸、葡庚糖酸、葡糖酸、谷氨酸、乙醇酸、羟萘酸、2-羟基乙磺酸、乳酸、马来酸、苹果酸、扁桃酸、甲磺酸、粘康酸、2-萘磺酸、丙酸、水杨酸、琥珀酸、二苯甲酰基-L-酒石酸、酒石酸、对甲苯磺酸、三甲基乙酸、三氟乙酸等形成的酸加成盐；和
(2)当母体化合物中存在的酸质子被金属离子，例如，碱金属离子(例如，Na +、K +或Li +)，碱土金属离子(如Ca 2+或Mg 2+)或铝离子代替；或者与有机碱或无机碱配位时形成的盐。可接受的有机碱包括二乙醇胺、乙醇胺、N-甲基葡糖胺、三乙醇胺、氨丁三醇等。可接受的无机碱包括氢氧化铝、氢氧化钙、氢氧化钾、碳酸钠和氢氧化钠。
在本发明中，术语“卤素”是指氟、溴、氯或碘原子。
术语“C 1-4烷基”是指包含1至4个碳原子的饱和的直链或支链的烃链。代表性的例子包括，但不限于，甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲 丁基、叔丁基基团。
术语“C 1-4亚烷基”指包含1至4个碳原子的二价烃链。代表性的例子包括，但不限于，CH 2-、-CH 2-CH 2-、-CH 2-CH 2-CH 2-、-CH 2-CH 2-CH 2-CH 2-等。
术语“C 2- 4烯基”是指具有至少一个双键并且具有2个至4个碳原子的直链或支链烃链。代表性的例子包括，但不限于，乙烯基、丙烯基、丁烯基等。
术语“C 2- 4炔基”是指具有至少一个三键并且具有2个至4个碳原子的直链或支链烃链。代表性的例子包括，但不限于，乙炔基、丙炔基、丁炔基等。
术语“C 1-4烷氧基”是指-O-(C 1-4烷基)，其中C 1-4烷基的定义如上所述。非限制性实施例包括甲氧基、乙氧基、丙氧基、丁氧基等。
术语“卤代C 1-4烷基”是指C 1-4烷基被一个或多个卤素取代，其中C 1-4烷基、卤素的定义如上所述。
术语“卤代C 1-4烷氧基”是指C 1-4烷氧基被一个或多个卤素取代，其中C 1-4烷氧基、卤素的定义如上所述。
术语“C 3-6环烷基”指包含3至6个碳原子的饱和或部分不饱和单环烃***，代表性的例子包括，但不限于，环己基、环戊基、环丁基、环丙基、环己烯基等。
术语“3-6元杂烷基”指包含3至6个环原子，其中1-3个环原子为选自氮、氧或S(O) m(其中m是是0、1或2)的杂原子的饱和或部分不饱和单环烃***，代表性的例子包括，但不限于，吡咯烷基、咪唑烷基、四氢呋喃基、四氢吡喃基、四氢噻吩基、二氢咪唑基、二氢呋喃基、二氢吡唑基、二氢吡咯基、哌啶基、哌嗪基、吗啉基等。
术语“羟基”是指-OH基团。
术语“硝基”指-NO 2。
术语“氨基”是指-NH 2。
术语“氰基”是指-CN。
术语“一个键”指用“—”表示的一个共价键。
术语“离去基”是指在亲核取代反应期间可以容易地被亲核试剂置换的化学基团，例如H原子、OH、卤素原子如氯原子或溴原子、或磺酸酯。磺酸酯可以特别地为甲磺酸酯(-OS(O 2)-CH 3)、三氟甲磺酸酯(-OS(O) 2-CF 3)或对甲苯磺酸酯(-OS(O) 2-(p-Me-C 6H 4))。
“任选”或“任选地”意味着随后所描述的事件或环境可以但不必发生，该说明包括该事件或环境发生或不发生的场合。例如，“任选被烷基取代的杂环基团”意味着烷基可以但不必须存在，该说明包括杂环基团被烷基取代的情形和杂环基团不被烷基取代的情形。
“取代的”指基团中的一个或多个氢原子，优选为最多5个，更优选为1～3个氢原子彼此独立地被相应数目的取代基取代。不言而喻，取代基仅处在它们的可能的化学位置，本领域技术人员能够在不付出过多努力的情况下确定(通过实验 或理论)可能或不可能的取代。例如，具有游离氢的氨基或羟基与具有不饱和(如烯属)键的碳原子结合时可能是不稳定的。
“药物组合物”表示含有一种或多种本文所述化合物或其生理学上/可药用的盐或前体药物与其他化学组分的混合物，以及其他组分例如生理学/可药用的载体和赋形剂。药物组合物的目的是促进对生物体的给药，利于活性成分的吸收进而发挥生物活性。
具体实施方式
通过阅读下列实施例，本领域技术人员将会更好地理解本发明。这些实施例仅用于解释本发明。
本发明实施例中未注明具体条件的实验方法，通常按照常规条件，或按照原料或商品制造厂商所建议的条件。未注明具体来源的试剂，为市场购买的常规试剂。
化合物的结构是通过核磁共振(NMR)或/和质谱(MS)来确定的。NMR化学位移(δ)以10 -6(ppm)的单位给出。测定溶剂为氘代氯仿(CDCl 3)，内标为四甲基硅烷(TMS)。使用下列缩写：s为单峰，bs为宽单峰，d为二重峰，t为三重峰，qdt为四重峰，m为多重峰或大量峰，dd为双二重峰等。
液质联用仪：Agilent LCMS1260/MSD6120，色谱柱：Agilent ZORBAX SB-C18，2.1*50mm，1.8μm，流动相：A：H 2O(0.1％FA)，B：乙腈，梯度洗脱，0.5mL/min，45.0℃，电离模式：API-ES，极性：正。
核磁共振仪：Bruker ARX-500型和Bruker ARX-400型。
MTT检测仪器：Thermo Scientific Multiskan GO全波长酶标仪。
薄层层析硅胶板使用青岛GF254硅胶板，薄层色谱法(TLC)使用的硅胶板采用的规格是0.15mm～0.2mm，薄层层析分离纯化产品采用的规格是0.4mm～0.5mm。
柱层析一般使用烟台黄海硅胶200～300目硅胶为载体。
实施例中如无特殊说明，反应均在氩气氛或氮气氛下进行。
实施例中如无特殊说明，反应中的溶液是指水溶液。
实施例中如无特殊说明，反应的温度为室温。
实施例中的反应进程的监测采用薄层色谱法(TLC)。
实施例1
4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8-二甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮1
Figure PCTCN2019124575-appb-000010
第一步
4-(苄胺基)-6-氯-烟酸甲酯102
将4,6-二氯烟酸甲酯101(6g，29.1mmol)，苄胺(3.4g，32mmol)和三乙胺(9.5g，87.3mmol)溶于30mL DMF中，室温搅拌过夜。TLC(石油醚：乙酸乙酯＝5:1) 检测，反应结束后，加入200mL水，搅拌30分钟，过滤，水洗，抽干。获得标题化合物，白色固体7.6g，收率94.1％。
MS(ESI):277.7[M+1] +。
第二步
(4-(苄氨基)-6-氯吡啶-3-基)甲醇103
将氢化铝锂(2.1g，54.9mmol)悬浮于70mL四氢呋喃中，冷却至-20℃下，加入4-(苄胺基)-6-氯-烟酸甲酯102(7.6g，27.4mmol)，在-20℃下反应2小时。TLC(石油醚：乙酸乙酯＝3:1)检测，反应结束后，依次加入2.1mL水、3.2mL 10％氢氧化钠溶液、10.2mL水，搅拌30分钟，干燥，过滤，旋干。获得标题化合物，白色固体7.5g，收率100％。
MS(ESI):249.7[M+1] +。
第三步
4-(苄胺基)-6-氯烟碱醛104
将(4-(苄氨基)-6-氯吡啶-3-基)甲醇103(7.5g，30.2mmol)溶于100mL二氯甲烷中，冷却至0℃下，分批加入DMP(戴斯-马丁氧化剂，15.3g，36.3mmol)，室温搅拌过夜。TLC(乙酸乙酯)检测，反应结束后，加入100mL 10％NaOH溶液搅拌30分钟，分液，二氯甲烷萃取，合并，干燥，旋干获得粗品。粗品经柱层析纯化(PE：EA＝3:1)，获得标题化合物，黄色油状物6.5g。收率87.1％。
MS(ESI):247.7[M+1] +。
第四步
N-苄基-2-氯-5-((4-乙基哌嗪-1-基)甲基)哌啶-4-胺105
将4-(苄胺基)-6-氯烟碱醛104(6.5g，26.3mmol)和N-乙基哌嗪溶于50mL二氯甲烷(3.6g，31.6mmol)中，冷却至0℃下，分批加入三乙酰氧基硼氢化钠(8.3g，39.45mmol)，室温搅拌过夜。TLC(乙酸乙酯)检测，反应结束后，加入100mL饱和碳酸氢钠溶液搅拌30分钟，分液，二氯甲烷萃取(100mL×3)，合并，干燥，旋干获得粗品。粗品经柱层析纯化(乙酸乙酯：甲醇＝10:1)，获得标题化合物，无色油状物7.8g。收率85.9％。
MS(ESI):348.5[M+1] +。
第五步
N 4-苄基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2,4-二胺106
在氮气下，将中间体N-苄基-2-氯-5-((4-乙基哌嗪-1-基)甲基)哌啶-4-胺105(6g，17.4mmol)、二苯甲酮亚胺(3.7g,20.8mmol)、叔丁醇钠(2.34g,24.36mmol)、Pd 2(dba) 3(1.6g,1.74mmol)和BINAP(3.24g,5.22mmol)溶于100mL甲苯中，在100℃下反应14小时。TLC(DCM：MeOH＝10:1)检测，反应结束后，加入100mL甲基叔丁基醚搅拌30分钟，过滤，洗涤，旋干获得粗品。将粗品溶于50mL THF和20mL甲醇中，加入10mL浓盐酸室温搅拌2小时。TLC(二氯甲烷：甲醇＝10:1) 检测，反应结束后，旋干反应液，加入饱和碳酸氢钠，乙酸乙酯萃取，保留水相，二氯甲烷萃取，合并有机相，干燥，旋干。获得标题化合物，白色固体3g，收率52.9％。
MS(ESI):326.2[M+1] +。
第六步
3-氨基丁酸甲酯108
在0℃下，将二氯亚砜(8.4mL,116.4mmol)滴入40mL甲醇中，搅拌1小时，3-氨基丁酸107(4g,38.8mmol)加入反应体系中，室温搅拌5小时。TLC(二氯甲烷：甲醇＝5:1)检测，反应结束后，旋干反应液，甲醇带干2次。获得标题化合物，无色油状物5g，收率100％。
第七步
3-((5-溴-3-氟-2-硝基苯基)氨基)丁酸甲酯109
将1,3-二氟-5溴-2-硝基苯(2g,8.4mmol)和3-氨基丁酸甲酯108(0.98g,8.4mmol)溶于20mL DMF中，加入DIEA(3,25g,25,2mmol)，室温搅拌过夜。TLC(石油醚：乙酸乙酯＝5:1)检测，反应结束后，加入水50mL水，乙酸乙酯萃取(3×25mL)，合并，水洗，干燥，旋干获得粗品。粗品经柱层析纯化(石油醚：乙酸乙酯＝50:1)，获得标题化合物，黄色固体2.4g，收率84.5％。
MS(ESI):336.1[M+1] +。
第八步
3-((2-氨基-5-溴-3氟苯基)氨基)丁酸甲酯110
将3-((5-溴-3-氟-2-硝基苯基)氨基)丁酸甲酯109(2g,5.9mmol)溶于15mL DMF中，加入氯化亚锡(6.7g,29.8mmol)，室温搅拌过夜。TLC(石油醚：乙酸乙酯＝5:1)检测，反应结束后，加入水50mL水，乙酸乙酯萃取(3×25mL)，合并，水洗，干燥，旋干。获得标题化合物，橙黄色油状物2.2g，收率100％。
MS(ESI):306.2[M+1] +。
第九步
3-(6-溴-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯111
将3-((2-氨基-5-溴-3氟苯基)氨基)丁酸甲酯110(2.2g,5.9mmol)溶于9mL原甲酸三甲酯和6mL醋酸中，在90℃下反应8小时。TLC(石油醚：乙酸乙酯＝1:1)检测，反应结束后，旋干反应液，加入饱和碳酸氢钠溶液，乙酸乙酯萃取(3×15mL)，合并，水洗，干燥，旋干获得粗品。粗品溶于10mL石油醚：乙酸乙酯＝1:10的溶液中，搅拌1小时，过滤，石油醚洗涤，抽干，获得标题化合物，白色固体1.2g，收率61.2％。
MS(ESI):330.1[M+1] +。
第十步
3-(6-(2-氯-5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯113
在氮气保护下，将3-(6-溴-4氟-2甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯111(2.5g,8mmol)、联硼酸频那醇酯(2.3g,9.1mmol)、醋酸钾(2.4g,24mmol)和Pd(dppf)Cl 2(326mg,0.4mmol)溶于70mL二氧六环中，在100℃下反应2小时。HPLC检测反应，反应结束后，将中间体2,4-二氯-5氟嘧啶(1.7g,10.4mmol)、碳酸铯(7.8g,24mmol)、Pd(dppf)Cl 2(326mg,0.4mmol)和7mL水加入反应液，在110℃下反应14小时。LC-MS检测反应，原料不再减少时，停止反应，旋干反应液，加入水，乙酸乙酯萃取(3×25mL)，合并，干燥，旋干获得粗品。粗品经柱层析纯化(石油醚：乙酸乙酯＝1:1)，获得标题化合物，黄色固体1g，收率32.5％。
MS(ESI):381.1[M+1] +。
第十一步
3-(6-(2-((4-(苄胺基)-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)-5-氟嘧啶-4-基)-4氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯114
在氮气保护下，将3-(6-(2-氯-5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯113(0.7g,1.8mmol)、N 4-苄基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2,4-二胺(0.72g,2.2mmol)、Pd 2(dba) 3(82mg,0.09mmol)、2-二环己基磷-2,4,6-三异丙基联苯(X-phos)(128mg，0.27mmol)和碳酸钾(0.745g，5.4mmol)悬浮于70mL二氧六环中，在100℃下反应12小时。TLC检测(二氯甲烷：甲醇＝5:1)，反应完全后，停止反应，旋干反应液，加入水，二氯甲烷萃取(3×25mL)，合并，干燥，旋干获得粗品。粗品经柱层析纯化(二氯甲烷：甲醇＝5:1)，获得标题化合物，棕黄色固体0.5g，收率38.9％。
MS(ESI):670.7[M+1] +。
第十二步
3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯115
在0℃下，将3-(6-(2-((4-(苄胺基)-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)-5-氟嘧啶-4-基)-4氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯114(0.5g，0.7mmol)溶于4mL浓硫酸中，搅拌30分钟。TLC检测(二氯甲烷：甲醇＝5:1)，反应完全后，停止反应，将反应液倒入碎冰中，用10％氢氧化钠调节pH＝8-9，乙酸乙酯萃取(3×20mL)，合并，干燥，旋干获得标题化合物0.4g，棕色固体，收率100％。
MS(ESI):580.6[M+1] +。
第十三步
3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸116
将3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯115(0.4g，6.9mmol)，溶于5mL甲醇和5mL10％氢氧化钠中，室温搅拌过夜。TLC检测(二氯甲烷：甲醇＝5:1)， 反应完全后，停止反应，用浓盐酸调节pH＝4-5，旋干，干燥，加入50mL乙醇搅拌5小时，过滤，洗涤，旋干，获得标题化合物，0.8g黄色固体，收率100％。
MS(ESI):566.6[M+1] +。
第十四步
4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8-二甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮1
将3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸116(0.8g，0.7mmol)、三乙胺(212mg，2.1mmol)溶于50mL二氯甲烷中，加入HATU(323mg，0.85mmol)，室温搅拌过夜。TLC检测(DCM：MeOH＝5:1)，反应完全后，停止反应，加入水，二氯甲烷萃取(3×20mL)，合并，干燥，旋干获得粗品。粗品经制备硅胶板(DCM：MeOH＝5:1)纯化，获得标题化合物，白色固体32mg。
MS(ESI):548.6[M+1] +。
1H NMR(400MHz,CDCl 3))δ10.41(s,1H),10.21(s,1H),9.73(s,1H),8.89–8.47(m,2H),8.12(s,1H),7.59(d,J＝11.8Hz,1H),5.23(s,1H),4.02(m,1H),3.65–3.35(m,2H),3.10–2.55(m,9H),2.47–2.08(m,5H),1.79(d,J＝6.4Hz,3H),1.03(t,J＝6.5Hz,3H)。
实施例2
(R)-4 5-((4-乙基哌嗪-1-基)甲基)-1 4,25-二氟-1 2,8-二甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮2
Figure PCTCN2019124575-appb-000011
Figure PCTCN2019124575-appb-000012
第一步
(R)-3-氨基丁酸甲酯202
在0℃下，二氯亚砜(5.1g,43mmol)滴入40mL甲醇中，搅拌1小时，(R)-3-氨基丁酸201(3g,29mmol)加入反应体系中，室温搅拌5小时。TLC(二氯甲烷：甲醇＝5:1)检测，反应结束后，旋干反应液，甲醇带干2次。获得标题化合物，无色油状物4.0g，收率100％。所得产品直接用于下一步反应。
第二步
(R)-3-((5-溴-3-氟-2-硝基苯基)氨基)丁酸甲酯203
将1,3-二氟-5溴-2-硝基苯(6.8g，29mmol)和(R)-3-氨基丁酸甲酯202(4.0g，29mmol)溶于50mL DMF中，加入DIEA(11.2g，87mmol)，室温搅拌过夜。TLC(石油醚：乙酸乙酯＝5:1)检测，反应结束后，加入水100mL水，乙酸乙酯萃取(3×25mL)，合并，水洗，干燥，旋干获得粗品。粗品经柱层析纯化(石油醚：乙酸乙酯＝50:1)，获得标题化合物，黄色固体9g，收率90.3％。
MS(ESI):335.2[M+1] +。
第三步
(R)-3-((2-氨基-5-溴-3氟苯基)氨基)丁酸甲酯204
将(S)-3-((5-溴-3-氟-2-硝基苯基)氨基)丁酸甲酯203(9g，26.8mmol)溶于50mL DMF中，加入氯化亚锡(30.3g，34mmol)，室温搅拌过夜。TLC(石油醚：乙酸乙酯＝5:1)检测，反应结束后，加入水100mL水，乙酸乙酯萃取(3×50mL)，合并，水洗，干燥，旋干。获得标题化合物，橙黄色油状物10g，收率100％。
MS(ESI):306.2[M+1] +。
第四步
(R)-3-(6-溴-4氟-2甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯205
将(S)-3-((2-氨基-5-溴-3氟苯基)氨基)丁酸甲酯204(10g，26.8mmol)溶于45mL原甲酸三甲酯和30mL醋酸中，在90℃下反应8小时。TLC(石油醚：乙酸乙酯＝1:1)检测，反应结束后，旋干反应液，加入饱和碳酸氢钠溶液，乙酸乙酯萃取(3×25mL)，合并，水洗，干燥，旋干获得粗品。粗品溶于20mL乙酸乙酯：石油醚＝1:10的溶液中，搅拌1小时，过滤，石油醚洗涤，抽干，获得标题化合物，白色固体6.2g，收率69.7％。
MS(ESI):330.1[M+1] +。
第五步
(R)-3-(6-(2-氯-5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯207
在氮气保护下，将(R)-3-(6-溴-4氟-2甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯205(5g，15.1mmol)、联硼酸频那醇酯(4.6g，18.2mmol)、醋酸钾(4.84g，45.3mmol)和Pd(dppf)Cl 2(0.64g，0.75mmol)溶于120mL二氧六环中，在100℃下反应2小时。HPLC检测反应，反应结束后，将中间体2,4-二氯-5氟嘧啶(3.2g，19.6mmol)、碳酸铯(14.7g，45.3mmol)、Pd(dppf)Cl 2(0.61g，0.7mmol)和12mL水加入反应液，在110℃下反应14小时。LC-MS检测反应，原料不再减少时，停止反应，旋干反应液，加入水，乙酸乙酯萃取(3×40mL)，合并，干燥，旋干获得粗品。粗品经柱层析纯化(石油醚：乙酸乙酯＝1:1)，获得标题化合物，黄色固体3.1g，收率53.6％。
MS(ESI):381.1[M+1] +。
第六步
(R)-3-(6-(2-((4-(苄胺基)-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)-5-氟嘧啶-4-基)-4氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯208
在氮气保护下，将(R)-3-(6-(2-氯-5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯207(1.1g，2.95mmol)、N 4-苄基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2,4-二胺(0.8g，2.46mmol)、Pd 2(dba) 3(112mg，0.12mmol)、X-phos(175mg，0.37mmol)和碳酸钾(1.02g，7.38mmol)悬浮于100mL二氧六环中，在100℃下反应12小时。TLC检测(二氯甲烷：甲醇＝5:1)，反应完全后，停止反应，旋干反应液，加入水，二氯甲烷萃取(3×30mL)，合并，干燥，旋干获得粗品。粗品经柱层析纯化(二氯甲烷：甲醇＝5:1)，获得标题化合物，棕黄色固体0.4g，收率20.3％。
MS(ESI):670.7[M+1] +。
第七步
(R)-3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯209
在0℃下，将(R)-3-(6-(2-((4-(苄胺基)-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)-5-氟嘧啶-4-基)-4氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯208(0.4g，0.6mmol)溶于4mL浓硫酸中，搅拌30分钟。TLC检测(二氯甲烷：甲醇＝5:1)，反应完全后，停止反应，将反应液倒入碎冰中，用10％氢氧化钠调节到pH＝8-9，乙酸乙酯萃取(3×30mL)，合并，干燥，旋干获得标题化合物0.4g，棕色固体，收率100％。
第八步
(R)-3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸210
将(R)-3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸甲酯209(0.4g，6.9mmol)，溶于5mL甲醇和5mL10％氢氧化钠中，室温搅拌过夜。TLC检测(二氯甲烷：甲醇＝5:1)，反应完全后，停止反应，用浓盐酸调节pH＝4-5，旋干，干燥，加入50mL乙醇搅拌5小时，过滤，洗涤，旋干，获得标题化合物，0.5g黄色固体，收率100％。
MS(ESI):566.6[M+1] +。
第九步
(R)-4 5-((4-乙基哌嗪-1-基)甲基)-1 4,25-二氟-1 2,8-二甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮2
将(R)-3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丁酸210(0.5g，0.69mmol)、三乙胺(212mg，2.1mmol)溶于50mL二氯甲烷中，加入HATU(323mg，0.85mmol)，室温搅拌过夜。TLC检测(二氯甲烷：甲醇＝5:1)，反应完全后，停止反应，加入水，二氯甲烷萃取(3×30mL)，合并，干燥，旋干获得粗品。粗品经制备硅胶板(二氯甲烷：甲醇＝5:1)纯化，获得标题化合物，白色固体24mg，收率：7.4％。
MS(ESI):548.6[M+1] +。
1H NMR(400MHz,DMSO)δ10.41(s,1H),10.21(s,1H),9.73(s,1H),8.89–8.47(m,2H),8.12(s,1H),7.59(d,J＝11.8Hz,1H),5.23(s,1H),3.85(d,J＝13.2Hz,1H),3.65–3.35(m,2H),3.10–2.55(m,9H),2.47–2.08(m,5H),1.79(d,J＝6.4Hz,3H),1.03(t,J＝6.5Hz,3H)。
实施例3
(S)-4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8-二甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮3
Figure PCTCN2019124575-appb-000013
依据实施例2中描述的方法，用(S)-3-氨基丁酸甲酯作为起始原料，获得(S)-4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8-二甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮3，白色固体。
MS(ESI):548.6[M+1] +。
1H NMR(400MHz,DMSO)δ10.41(s,1H),10.21(s,1H),9.73(s,1H),8.89–8.47(m,2H),8.12(s,1H),7.59(d,J＝11.8Hz,1H),5.23(s,1H),3.85(d,J＝13.2Hz,1H),3.65–3.35(m,2H),3.10–2.55(m,9H),2.47–2.08(m,5H),1.79(d,J＝6.4Hz,3H),1.03(t,J＝6.5Hz,3H)。
实施例4
4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2-甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮4
Figure PCTCN2019124575-appb-000014
Figure PCTCN2019124575-appb-000015
第一步
氨基丁酸甲酯402
在0℃下，将二氯亚砜(10g,84mmol)滴入40mL甲醇中，搅拌1小时，氨基丁酸(5g,56mmol)加入反应体系中，室温搅拌5小时。TLC(DCM：MeOH＝5:1)检测，反应结束后，旋干反应液，MeOH带干2次。获得标题化合物，无色油状物6g，收率100％。所得产品直接用于下一步反应。
第二步
3-((5-溴-3-氟-2-硝基苯基)氨基)丙酸甲酯403
将1,3-二氟-5溴-2-硝基苯402(13.3g(56mmol)和3-氨基丙酸甲酯(6g(56mmol)溶于100mL DMF中，加入DIEA(21.6g(168mmol)，室温搅拌过夜。TLC(PE：EA＝5:1)检测，反应结束后，加入水150mL水，等体积乙酸乙酯萃取2次，合并有机相，水洗，干燥，旋干获得粗品。粗品经柱层析纯化(PE：EA＝50:1)，获得标题化合物，橙黄色固体14g，收率77.8％。
MS(ESI):322.1[M+1] +。
第三步
3-((2-氨基-5-溴-3-氟苯基)氨基)丙酸甲酯404
将3-((5-溴-3-氟-2-硝基苯基)氨基)丙酸甲酯403(14g,43.6mmol)溶于150mL DMF中，加入氯化亚锡(49g,218mmol)，室温搅拌过夜。TLC(石油醚：乙酸乙酯＝5:1)检测，反应结束后，加入水500mL水，乙酸乙酯萃取(50ml×3)，合并有机相，水洗，干燥，旋干。获得标题化合物，白黄色油状物12g，收率100％。
MS(ESI):292.2[M+1] +。
第四步
3-(6-溴-4氟-2甲基-1H-苯并[d]咪唑-1-基)丙酸甲酯405
将3-((2-氨基-5-溴-3氟苯基)氨基)丙酸甲酯404(12g,41mmol)溶于90mL原乙酸三甲酯和54mL醋酸中，在90℃下反应8小时。TLC(石油醚：乙酸乙酯＝1:1)检测，反应结束后，旋干反应液，加入约100ml饱和碳酸氢钠溶液，乙酸乙酯萃取(50ml×3)，合并有机相，水洗，干燥，旋干获得粗品。粗品溶于10mL乙酸乙酯：石油醚＝1:10的溶液中，搅拌1小时，过滤，石油醚洗涤，抽干，获得标题化合物，白色固体6.8g，收率52.3％。
MS(ESI):316.1[M+1] +。
第五步
3-(6-(2-氯-5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丙酸甲酯407
在氮气保护下，将3-(6-溴-4氟-2甲基-1H-苯并[d]咪唑-1-基)丙酸甲酯405(5g,15.9mmol)、联硼酸频那醇酯(4.8g,19.1mmol)、醋酸钾(4.7g,47.7mmol)和Pd(dppf)Cl 2(650mg,0.8mmol)溶于100mL二氧六环中，在100℃下反应2小时。HPLC检测反应，反应结束后，将中间体2,4-二氯-5氟嘧啶(3.4g,20.4mmol)、碳酸铯(15.5g,47.7mmol)、Pd(dppf)Cl 2(650mg,0.8mmol)和15mL水加入反应液，在110℃下反应14小时。LC-MS检测反应，原料不再减少时，停止反应，旋干反应液，加入约100ml水，乙酸乙酯萃取(50ml×3)，合并有机相，干燥，旋干获得粗品。粗品经硅胶柱层析纯化(石油醚：乙酸乙酯＝1:1)，获得标题化合物，棕色固体2.8g，收率48.2％。
MS(ESI):367.1[M+1] +。
第六步
3-(6-(2-((4-(苄胺基)-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)-5-氟嘧啶-4-基)-4氟-2-甲基-1H-苯并[d]咪唑-1-基)丙酸甲酯408
在氮气保护下，将3-(6-(2-氯-5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丙酸甲酯407(2.8g，7.6mmol)、N 4-苄基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2,4-二胺(2g，6.3mmol)、Pd 2(dba) 3(580mg，0.63mmol)、X-phos(900mg，1.89mmol)和碳酸钾(2.6g，18.9mmol)悬浮于70mL二氧六环中，在100℃下反应12小时。TLC检测(DCM：MeOH＝5:1)，反应完全后，停止反应，旋干反应液，加入约70ml水，乙酸乙酯萃取(50ml×3)，合并有机相，干燥，旋干获得粗品。粗品经 硅胶柱层析纯化(DCM：MeOH＝5:1)，获得标题化合物，棕色油状物1.5g，收率30.9％。
MS(ESI):656.7[M+1] +。
第七步
3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丙酸甲酯409
在0℃下，将3-(6-(2-((4-(苄胺基)-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)-5-氟嘧啶-4-基)-4氟-2-甲基-1H-苯并[d]咪唑-1-基)丙酸甲酯408(1.5g，2.1mmol)溶于4mL浓硫酸中，搅拌30分钟。TLC检测(DCM：MeOH＝5:1)，反应完全后，停止反应，将反应液倒入约100g碎冰中，用10％氢氧化钠条件pH＝8-9，乙酸乙酯萃取(50ml×3)，合并有机相，干燥，旋干获得标题化合物0.9g，棕色固体，收率63.6％。
MS(ESI):566.6[M+1] +。
第八步
3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丙酸410
将3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丙酸甲酯409(0.9g，1.6mmol)，溶于5mL甲醇和5mL 10％氢氧化钠中，室温搅拌过夜。TLC检测(DCM：MeOH＝5:1)，反应完全后，停止反应，用浓盐酸调节pH＝4-5，旋干，干燥，加入50mL乙醇搅拌5小时，过滤，洗涤，滤液旋干，获得标题化合物，1.6g灰白色固体，收率100％。
MS(ESI):552.6[M+1] +。
第九步
4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2-甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮4
将3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)哌啶-2-基)氨基)5-氟嘧啶-4-基)-4-氟-2-甲基-1H-苯并[d]咪唑-1-基)丙酸410(1.6g，0.9mmol)、三乙胺溶(270mg，2.7mmol)于50mL二氯甲烷中，加入HATU(510mg，1.3mmol)，室温搅拌过夜。TLC检测(DCM：MeOH＝5:1)，反应完全后，停止反应，加入约50ml水，分液，乙酸乙酯萃取(50ml×3)，干燥，旋干获得粗品。粗品经制备硅胶板(DCM：MeOH＝5:1)纯化，获得标题化合物，白色固体35mg。
MS(ESI):534.6[M+1]+。
1H NMR(400MHz,DMSO-d 6)δ10.68(s,1H),10.11(s,1H),9.52(s,1H),8.67(d,J＝4.1Hz,1H),8.39(s,1H),8.08(s,1H),7.58(d,J＝11.8Hz,1H),4.71(s,2H),3.97–3.41(m,2H),3.03–2.60(m,9H),2.48–1.93(m,6H),1.02(s,3H).
实施例5
4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8-二甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃5
Figure PCTCN2019124575-appb-000016
在4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8-二甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮1(100mg,0.182mmol)的THF(5mL)溶液中，在0℃下加入LiAlH4(10mg,0.26mmol)。室温搅拌过夜。TLC检测(DCM：MeOH＝5:1)，反应完全后，停止反应，在0℃下加入约10ml饱和氯化铵水溶液，分液，乙酸乙酯萃取(10ml×3)，干燥，旋干获得粗品。粗品经制备硅胶板(DCM：MeOH＝5:1)纯化，获得标题化合物，白色固体60mg。
MS(ESI):534.2[M+1]+。
1H NMR(400MHz,CDCl 3))δ10.41(s,1H),10.21(s,1H),9.73(s,1H),8.89–8.47(m,2H),8.12(s,1H),7.59(d,J＝11.8Hz,1H),5.23(s,1H),3.85(d,J＝13.2Hz,1H),3.32–3.20(m,2H),3.10–2.55(m,9H),2.47–2.08(m,5H),2.00–1.84(m,2H),1.79(d,J＝6.4Hz,3H),1.03(t,J＝6.5Hz,3H)。
实施例7
4 5-((4-乙基哌嗪-1-基)甲基)-2 5-氟-8-甲基-1 1H-3,5-二氮杂-1(6,1)-吲哚-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮7
Figure PCTCN2019124575-appb-000017
Figure PCTCN2019124575-appb-000018
第一步
3-(6-溴-1H-吲哚-1-基)丁酸甲酯7-1
6-溴吲哚(1.0g,5.1mmol)、3-溴丁酸甲酯(1.38g,7.65mmol)、氢氧化钠(0.43g,7.65mmol)中加入N,N-二甲基甲酰胺(DMF,10mL),在氮气保护下70℃反应10小时。反应液中加入水(50mL)/乙酸乙酯(50mL)搅拌萃取，有机相干燥、过滤、浓缩后过硅胶柱(洗脱剂：乙酸乙酯/石油醚：1/30-1/10)得白色固体7-1(1.2g,收率80％。MS(ESI):296.1[M+1] +。
第二步
3-(6-(2-氯-5-氟嘧啶-4-基)-1H-吲哚-1-基)丁酸甲酯7-3
在氮气保护下，将3-(6-溴-1H-吲哚-1-基)丁酸甲酯7-1(2.4g,8mmol)、联硼酸频那醇酯(2.3g,9.1mmol)、醋酸钾(2.4g,24mmol)和Pd(dppf)Cl 2(326mg,0.4mmol)溶于70mL二氧六环中，在100℃下反应2小时。HPLC检测反应，反应结束后，将中间体2,4-二氯-5氟嘧啶(1.7g,10.4mmol)、碳酸铯(7.8g,24mmol)、Pd(dppf)Cl 2(326mg,0.4mmol)和7mL水加入反应液，在110℃下反应14小时。LC-MS检测反应，原料不再减少时，停止反应，旋干反应液，加入水，乙酸乙酯萃取(3×25mL)，合并，干燥，旋干获得粗品。粗品经柱层析纯化(石油醚：乙酸乙酯＝1:1)，获得标题化合物，黄色固体1g，收率32.5％。
MS(ESI):348.1[M+1] +。
第三步
3-(6-(2-((4-(苄氨基)-5-((4-乙基哌嗪-1-基)甲基)吡啶-2-基)氨基)-5-氟嘧啶-4-基)-1H-吲哚-1-基)丁酸甲酯7-4
在氮气保护下，将3-(6-(2-氯-5-氟嘧啶-4-基)-1H-吲哚-1-基)丁酸甲酯7-3(0.63g,1.8mmol)、N 4-苄基-5-((4-乙基哌嗪-1-基)甲基)吡啶-2,4-二胺(0.72g,2.2mmol)、Pd 2(dba) 3(82mg,0.09mmol)、2-二环己基磷-2,4,6-三异丙基联苯(X-phos)(128mg，0.27mmol)和碳酸钾(0.745g，5.4mmol)悬浮于70mL二氧六环中，在100℃下反应12小时。TLC检测(二氯甲烷：甲醇＝5:1)，反应完全后，停止反应，旋干反应液，加入水，二氯甲烷萃取(3×25mL)，合并，干燥，旋干获得粗品。粗品经柱层析纯化(二氯甲烷：甲醇＝5:1)，获得标题化合物，棕黄色固体0.45g，收率40％。
MS(ESI):637.1[M+1] +。
第四步
3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)吡啶-2-基)氨基)-5-氟嘧啶-4-基)-1H-吲哚-1-基)丁酸甲酯7-5
在0℃下，将3-(6-(2-((4-(苄氨基)-5-((4-乙基哌嗪-1-基)甲基)吡啶-2-基)氨基)-5-氟嘧啶-4-基)-1H-吲哚-1-基)丁酸甲酯7-4(0.45g，0.7mmol)溶于4mL浓硫酸中，搅拌30分钟。TLC检测(二氯甲烷：甲醇＝5:1)，反应完全后，停止反应，将反应液倒入碎冰中，用10％氢氧化钠调节pH＝8-9，乙酸乙酯萃取(3×20mL)，合并，干燥，旋干获得标题化合物0.4g，棕色固体，收率100％。
MS(ESI):547.1[M+1] +。
第五步
3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)吡啶-2-基)氨基)-5-氟嘧啶-4-基)-1H-吲哚-1-基-)丁酸7-6
将3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)吡啶-2-基)氨基)-5-氟嘧啶-4-基)-1H-吲哚-1-基)丁酸甲酯7-5(0.38g，6.9mmol)，溶于5mL甲醇和5mL10％氢氧化钠中，室温搅拌过夜。TLC检测(二氯甲烷：甲醇＝5:1)，反应完全后，停止反应，用浓盐酸调节pH＝4-5，旋干，干燥，加入50mL乙醇搅拌5小时，过滤，洗涤，旋干，获得标题化合物，0.38g黄色固体，收率100％。
MS(ESI):551.1[M+1] +。
第六步
4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8-二甲基-1 1H-3,5-二氮杂-1(6,1)-吲哚-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮7
将3-(6-(2-((4-氨基-5-((4-乙基哌嗪-1-基)甲基)吡啶-2-基)氨基)-5-氟嘧啶-4-基)-1H-吲哚-1-基)丁酸7-6(0.38g，0.7mmol)、三乙胺(212mg，2.1mmol)溶于50mL二氯甲烷中，加入HATU(323mg，0.85mmol)，室温搅拌过夜。TLC检测(DCM：MeOH＝5:1)，反应完全后，停止反应，加入水，二氯甲烷萃取(3×20mL)，合并，干燥，旋干获得粗品。粗品经制备硅胶板(DCM：MeOH＝5:1)纯化，获得标题化合物7，白色固体120mg。
MS(ESI):515.2[M+1] +。
1H NMR(400MHz,CDCl 3))δ10.23(s,1H),10.01(s,1H),9.21(s,1H),8.45–8.20(m,2H),8.12(d,J＝11.8Hz,1H),7.82(s,1H),7.59(d,J＝11.8Hz,1H),7.33(d,J＝12.1Hz,1H),6.41(d,J＝12.1Hz,1H),5.23(s,1H),4.5(br,1H),4.06(m,1H),3.65(s,2H),2.84–2.62(m,2H),2.47–2.08(m,8H),1.68(d,J＝6.2Hz,3H),1.01(t,J＝6.5Hz,3H)。
实施例8
4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8,8-三甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮8
Figure PCTCN2019124575-appb-000019
依据实施例1中描述的方法，将第六步中的反应原料3-氨基丁酸107换为3-氨基-3-甲基丁酸，获得4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8,8-三甲基-1 1H-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-6-酮8，白色固体。
MS(ESI):562.2[M+1] +。
1H NMR(400MHz,CDCl 3))δ10.31(s,1H),10.01(s,1H),9.73(s,1H),8.89–8.47(m,2H),8.12(s,1H),7.62(d,J＝11.8Hz,1H),3.75(s,2H),3.35(q,J＝6.5Hz,2H),2.64(s,2H),2.51(s,3H),2.47–2.35(m,8H),1.43(s,6H),1.03(t,J＝6.5Hz,3H)。
实施例9
5'-((4-乙基哌嗪-1-基)甲基)-4',5'-二氟-2'-甲基螺[环丙烷-1,8'-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃]-6'-酮9
Figure PCTCN2019124575-appb-000020
依据实施例1中描述的方法，将第六步中的反应原料3-氨基丁酸107换为2-(1-氨基环丙基)乙酸，获得5'-((4-乙基哌嗪-1-基)甲基)-4',5'-二氟-2'-甲基螺[环丙烷-1,8'-3,5-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃]-6'-酮9，白色固体。
MS(ESI):560.1[M+1] +。
1H NMR(400MHz,CDCl 3))δ10.34(s,1H),10.24(s,1H),9.71(s,1H),8.89–8.47(m,2H),8.12(s,1H),7.62(d,J＝11.8Hz,1H),3.75(s,2H),3.35(q,J＝6.5Hz,2H),2.84(s,2H),2.62(s,3H),2.47–2.35(m,8H),1.03(t,J＝6.5Hz,3H),0.76-0.74(m,2H),0.52-0.48(m,2H)
实施例6、10和11
依据实施例1-3中描述的方法，用适当的化合物作为起始原料，获得如下化合物：
4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8-二甲基-1 1H-5-氧杂-3-氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃6；
4 5-((4-乙基哌嗪-1-基)甲基)-1 4,2 5-二氟-1 2,8-二甲基-1 1H-3,6-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-5-酮10；
4 5-(4-乙基哌嗪-1-基)-1 4,2 5-二氟-1 2,8-二甲基-1 1H-3,6-二氮杂-1(6,1)-苯并[d]咪唑-2(4,2)-嘧啶-4(2,4)-吡啶环辛蕃-5-酮11。
测试例1
CDK4/Cyclin D1抑制作用的测定
关于CDK4/Cyclin D1的IC 50测定法是按如下步骤进行的。使用96孔过滤平板(Millipore MADVN6550)。总体积为0.1ml，含有缓冲液A(20mM TRIS(三[羟甲基]氨基甲烷)(pH7.4)，50mM NaCl，1mM二硫苏糖醇，10mM MgCl2)、25μM ATP(含有0.25μCi[32P]ATP)、20ng CDK4、1μg视网膜母细胞瘤蛋白和供试化合物的适当比例缓冲液A稀释液。采用没有加入供试化合物的单独缓冲液A作为没有抑制作用的对照。使用含有过量EDTA的缓冲液A测定在没有酶活性存在下的背景32P水平。向小孔加入除ATP以外的所有组分，将平板置于平板混合机上达2分钟。加入[32P]ATP引发反应，将平板在25℃下孵育15分钟。加入0.1ml 20％三氯乙酸(TCA)终止反应。使平板在4℃下保持至少1小时，以便底物沉淀出来。然后将小孔用0.2ml 10％TCA洗涤5次，利用β平板计数器(wallac Inc.，Gaithersburg，MD)测定32P的结合。利用中位效应法测定供试化合物的IC 50(Chou,T-C and Talalay P.Applications of the median effect principle for the assessmentof low-dose risk of carcinogens and for the quantitation ofsynergism and antagonism of chemotherapeutic agents.In New Avenues in Developmental Cancer Chemotherapy(Eds.Harrap,K.T.and Connors,T.A.),pp.37-64.Academic Press,New York,1987)。
实验方法：采用Caliper迁移率(Mobility Shift)方法进行CDK4/6激酶的抑制活性测定
1、1倍激酶缓冲液的配制：
1)、1倍CDK4激酶缓冲液的配制
分别取800μL母液浓度为1000mM的pH 7.5的HEPES、40μL母液浓度为 10％的Triton X-100，加入到39160μL的超纯水中，混匀。
2)、1倍CDK6激酶缓冲液的配制
分别取50mL母液浓度为1000mM的pH 7.5的HEPES、50μL母液浓度为30％的Brij-35，加入到949.95mL的超纯水中，混匀。
2、终止液的配制
分别取25mL母液浓度为4％的包被液Coating Reagent#3(Caliper仪器所使用的12-吸头芯片(sipper chip)中自带)、50mL母液浓度为1000mM pH 7.5的HEPES、50mL母液浓度为0.5M的EDTA、0.25mL母液浓度为30％的Brij-35，加入到374.75mL的超纯水中，混匀。
3、2.5倍激酶溶液的配制
1)、2.5倍CDK4/D3激酶溶液的配制
分别取7μL CDK4/D3酶溶液、9μL母液浓度为1M的DTT，加入到1784μL 1倍CDK4激酶缓冲液中，混匀。
2)、2.5倍CDK6/D3激酶溶液的配制
分别取18μL CDK6/D3酶溶液、14μL母液浓度为1M的DTT，加入到2768μL 1倍CDK6激酶缓冲液中，混匀。
4、2.5倍多肽溶液的配制
1)、2.5倍CDK4/D3多肽溶液的配制
分别取10μL母液浓度为100mM的ATP溶液、45μL母液浓度为1M的MgCl 2，45μL FAM标记的多肽8，加入到1700μL 1倍CDK4激酶缓冲液，混匀。
2)、2.5倍CDK6/D3多肽溶液的配制
分别取23μL母液浓度为100mM的ATP溶液、75μL母液浓度为1M的MgCl 2，75μLFAM标记的多肽8，加入到2827μL 1倍CDK6激酶缓冲液，混匀。
5、5倍测试物溶液配制：
取10mM测试物的DMSO储备液，用DMSO稀释制成浓度为50μM的溶液，作为母液。用DMSO将上述母液四倍逐级稀释制成12.5μM，3.125μM，0.78μM，0.195μM，0.0488μM，12.2nM，3nM，0.76nM，0.19nM的溶液，然后每个浓度分别用1倍激酶缓冲液稀释10倍，制成5倍化合物溶液。
6、CDK4/6酶学反应：
1)、384孔板中相对应的孔中分别加入5μL配制好的5倍测试物溶液、10μL配制好的2.5倍激酶溶液，室温孵育10分钟。
2)、相对应的孔中再分别加入10μL配制好的2.5倍多肽溶液，启动酶反应，28℃孵育5小时。
7、酶学检测：
每个相对应的孔中分别加入25μL终止液，终止反应。
8、Caliper仪器读取数据，并通过下式计算抑制率，然后采用GraphPad5.0软 件进行曲线拟合，得出IC 50值。
Figure PCTCN2019124575-appb-000021
测试例2
本发明化合物的体外细胞学抑制活性
实验方法：采用BrdU方法(BrdU细胞增殖试验试剂盒，Cell Signaling Technology公司)进行细胞增殖检测
1、试剂和化合物配制
1倍洗液配制：
将母液浓度为20倍的洗液用超纯水稀释成1倍洗液。
1倍检测抗体溶液配制：
将母液浓度为100倍的BrdU检测抗体用检测抗体稀释液稀释成1倍检测抗体溶液。
1倍HRP标记的二抗溶液配制：
将母液浓度为100倍的抗小鼠IgG，HRP标记抗体用HRP标记抗体稀释液稀释成1倍HRP标记的二抗溶液。
10倍BrdU溶液：
将母液浓度为1000倍的BrdU溶液用细胞对应的培养基稀释成10倍BrdU溶液。
配制测试化合物：
配制测试化合物母液：用100％DMSO配制成10mM的母液。
配制测试化合物梯度稀释溶液：取10mM的测试化合物母液用DMSO 4倍连续梯度稀释，浓度分别为2.5mM，625μM，156μM，39μM，9.8μM，2.5μM。分别取2μL的DMSO稀释的化合物加到198μL含10％FBS的培养液中配制成10倍测试物，测试物最高浓度为100μM，DMSO浓度为1％，共7个浓度梯度。
培养基配制：
MCF-7培养基：DMEM+10％FBS+0.01mg/mL胰岛素
2、试验步骤
(1)胰酶消化生长至80％的细胞(对数生长期)，离心收集细胞。用不含FBS的培养基重悬MDA-MB-435S和U87MG细胞，计数并调整接种96孔板，MDA-MB-435S细胞接种3000个/孔/81μL，U87MG细胞接种4000个/孔/81μL；用含1％FBS的培养基重悬MCF-7细胞，计数并调整接种96孔板，接种4000个/孔/82μL，置于37℃细胞培养箱中培养；
(2)培养24小时后MDA-MB-435S和U87MG细胞每孔添加FBS(9μL)，MCF-7细胞每孔添加8μL FBS，使FBS终浓度为10％；
(3)每孔加入不同浓度的10倍测试物(10μL)，使测试物终浓度分别为10μM，2.5μM，625nM，156nM，39nM，9.8nM，2.5nM，3复孔/组，37℃培养72小时；
溶剂对照：0.1％DMSO
空白对照：只加培养基，没有细胞
正常细胞对照：没有任何处理的正常细胞
(4)每孔加入10倍BrdU溶液(10μL)，细胞培养箱中孵育4小时后弃去培养基；
(5)每孔加入固定/变性液(10μL)，室温孵育30分钟，弃去溶液；
(6)每孔加入1倍检测抗体溶液(100μL)，室温孵育1小时，弃去溶液，用1倍洗液200μL/孔洗3次；
(7)每孔加入1倍HRP标记的二抗溶液(100μL)，室温孵育30分钟，弃去溶液，用1倍洗液200μL/孔洗3次；
(8)每孔加入TMB底物溶液(100μL)，室温孵育30分钟；
(9)每孔加入终止液(100μL)，酶标仪450nm检测OD值。
3、数据处理
1)细胞存活率(％)＝(OD 测试物-OD 空白对照)/(OD 正常细胞对照-OD 空白对照)×100％，
OD 空白对照：空白对照值，OD 正常细胞对照：正常细胞对照值；
2)数据使用GraphPad Prism 5软件处理，得到曲线及IC 50值。
Figure PCTCN2019124575-appb-000022

Claims (15)

  1. 一种通式(I)所示的化合物：
    Figure PCTCN2019124575-appb-100001
    或其互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体、或其混合物形式，或其药学上可接受的盐，
    其中：
    A和B各自独立地选自CH和N原子；
    U、V、W、X和Y各自独立地选自CH和N原子；
    Q选自一个键、C 1-4亚烷基和-C(＝O)-，所述C 1-4亚烷基任选被选自C 1-4烷基、卤代C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
    G选自一个键、-O-、-N(R 7)-、-C(＝O)-、C 1-4亚烷基、C 2-4烯基、-S-、-SO-、-SO 2-和3-6元杂环基，其中所述C 1-4亚烷基、C 2-4烯基和3-6元杂环基各自独立地任选被选自C 1-4烷基、卤代C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
    J选自一个键、-O-、-N(R 7)-、-C(＝O)-、C 1-4亚烷基、C 2-4烯基、-S-、-SO-、-SO 2-和3-6元杂环基，其中所述C 1-4亚烷基、C 2-4烯基和3-6元杂环基各自独立地任选被选自C 1-4烷基、卤代C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
    L选自一个键、-C(＝O)-、C 1-4亚烷基、C 2-4烯基、-S-、-SO-、-SO 2-和3-6元杂环基，其中所述C 1-4亚烷基、C 2-4烯基和3-6元杂环基各自独立地任选被选自C 1-4烷基、卤代C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
    R 1选自H原子、C 1-4烷基、氰基、C 2-4烯基和C 2-4炔基，其中所述C 1-4烷基、C 2-4烯基和C 2-4炔基各自独立地任选被选自卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
    R 2选自H原子和C 1-4烷基，其中所述C 1-4烷基任选被选自卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
    或者，R 1和R 2与其相连接的原子一起形成C 3-6环烷基或3-6元杂环基，所述 C 3-6环烷基和3-6元杂环基各自独立地任选被选自C 1-4烷基、卤代C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；
    R 3和R 6各自独立地选自H原子和C 1-4烷基，其中所述烷基任选被选自C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代
    R 4和R 5各自独立地选自H原子、C 1-4烷基、卤素、羟基和C 1-4烷氧基，其中所述C 1-4烷基和C 1-4烷氧基各自独立地任选被选自卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代；和
    R 7选自H原子和C 1-4烷基，其中所述烷基任选被选自C 1-4烷氧基、卤代C 1-4烷氧基、卤素、氨基、硝基、羟基和氰基中的一个或多个基团所取代。
  2. 根据权利要求1所述的通式(I)所示的化合物，其中-G-J-L-选自-NH-CH 2-CH 2-、-O-CH 2-CH 2-、-NH-C(＝O)-CH 2-和-C(＝O)-NH-CH 2-。
  3. 根据权利要求1或2所述的通式(I)所示的化合物，其中Q为一个键或亚甲基。
  4. 根据权利要求1-3中任一项所述的通式(I)所示的化合物，其中V、X和Y各自独立地为CH。
  5. 根据权利要求1-4中任一项所述的通式(I)所示的化合物，其中W为N原子。
  6. 根据权利要求1-5中任一项所述的通式(I)所示的化合物，其中R 1和R 2各自独立地选自H原子和C 1-4烷基，或者，R 1和R 2与其相连接的原子一起形成C 3-6环烷基。
  7. 根据权利要求1-6中任一项所述的通式(I)所示的化合物，其中R 3和R 6各自独立地为C 1-4烷基。
  8. 根据权利要求1-7中任一项所述的通式(I)所示的化合物，其中R 4和R 5各自独立地为卤素，优选F原子。
  9. 根据权利要求1-8中任一项所述的通式(I)所示的化合物，其选自：
    Figure PCTCN2019124575-appb-100002
  10. 一种制备根据权利要求1-9中任一项所述的通式(I)所示的化合物的方法，其包括：
    Figure PCTCN2019124575-appb-100003
    通式(IA)化合物在缩合剂存在下发生分子内成环反应形成通式(I)化合物，
    其中：
    LG 1和LG 2各自独立地为离去基；
    A、B、U、V、W、X、Y、G、J、L、R 1至R 6如权利要求1中所定义。
  11. 根据权利要求10所述的方法，其中所述缩合剂选自2-(7-氧化苯并三氮 唑)-N,N,N',N'-四甲基脲六氟磷酸酯、1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐、N,N'-二环己基碳化二亚胺、N,N'-二异丙基碳二酰亚胺、O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸酯、1-羟基苯并***、1-羟基-7-偶氮苯并三氮唑、O-苯并三氮唑-N,N,N',N'-四甲脲六氟磷酸酯、2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯、苯并三氮唑-1-基氧基三(二甲基氨基)磷鎓六氟磷酸盐和六氟磷酸苯并***-1-基-氧基三吡咯烷基磷，优选2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯；
    LG 1和LG 2各自独立地选自H原子、OH、卤素、甲磺酸酯、三氟甲磺酸酯和对甲苯磺酸酯；优选地，LG 1和LG 2各自独立地选自H原子、OH、Cl原子和Br原子；更优选地，LG 1为H原子，LG 2为OH原子。
  12. 根据权利要求10或11所述的方法，其中G为-NH-，LG 1为H原子，J为-C(＝O)-，且-LG 2为OH原子。
  13. 一种药物组合物，其包含治疗有效量的根据权利要求1-9中任一项所述的通式(I)所示的化合物以及药学上可接受的载体、稀释剂或赋形剂，优选地，所述药物组合物还包含一种或多种额外的抗肿瘤剂、抗炎剂、免疫抑制剂和/或免疫检测点抑制剂。
  14. 根据权利要求1-9中任一项所述的通式(I)所示的化合物或根据权利要求13所述的药物组合物在制备治疗CDK相关病症的药物中的用途，优选地，所述CDK相关病症选自癌症、炎症、病毒感染、心脏肥大和HIV。
  15. 根据权利要求14所述的用途，其中所述癌症选自膀胱癌、头颈癌、乳腺癌、胃癌、卵巢癌、结肠癌、肺癌、脑癌、喉癌、淋巴***癌、造血***癌、泌尿生殖道癌、胃肠癌、卵巢癌、***癌、胃癌、骨癌、小细胞肺癌、神经胶质瘤、结肠直肠癌和胰腺癌；
    所述炎症与类风湿性关节炎、狼疮、I型糖尿病、糖尿病性肾病变、多发性硬化、肾小球肾炎、慢性炎症和器官移植排斥有关；
    所述病毒感染与HIV病毒、人***状瘤病毒、疱疹病毒、痘病毒、EB病毒、新培斯病毒或腺病毒相关。
PCT/CN2019/124575 2018-12-19 2019-12-11 作为cdk抑制剂的大环化合物、其制备方法及其在医药上的应用 WO2020125513A1 (zh)

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JP2021533362A JP2022517723A (ja) 2018-12-19 2019-12-11 Ｃｄｋ阻害剤としての大環状化合物、その製造方法及びその医薬品における応用
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