WO2020151566A1 - 抑制pge2/ep4信号传导的化合物、其制备方法及其在医药上的应用 - Google Patents

抑制pge2/ep4信号传导的化合物、其制备方法及其在医药上的应用 Download PDF

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WO2020151566A1
WO2020151566A1 PCT/CN2020/072487 CN2020072487W WO2020151566A1 WO 2020151566 A1 WO2020151566 A1 WO 2020151566A1 CN 2020072487 W CN2020072487 W CN 2020072487W WO 2020151566 A1 WO2020151566 A1 WO 2020151566A1
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alkyl
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compound
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alkoxy
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French (fr)
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邓永奇
孙健
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Camphor Pharmaceuticals Ltd
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Camphor Pharmaceuticals Ltd
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Priority to CN202080002090.1A priority Critical patent/CN111989311B/zh
Priority to CA3126484A priority patent/CA3126484A1/en
Priority to AU2020212111A priority patent/AU2020212111B2/en
Priority to JP2021541481A priority patent/JP7488269B2/ja
Priority to KR1020217024124A priority patent/KR102861190B1/ko
Priority to ES20744659T priority patent/ES2995259T3/es
Priority to EP20744659.2A priority patent/EP3889134B1/en
Priority to US17/423,248 priority patent/US12240812B2/en
Publication of WO2020151566A1 publication Critical patent/WO2020151566A1/zh
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Definitions

  • the present invention relates to the field of medicine, and relates to a compound represented by general formula (I), a preparation method thereof, a pharmaceutical composition containing the derivative, and its use as a therapeutic agent, especially as a PGE2/EP4 signaling inhibitor in treatment Use in cancer and chronic inflammatory diseases.
  • a compound represented by general formula (I) a preparation method thereof, a pharmaceutical composition containing the derivative, and its use as a therapeutic agent, especially as a PGE2/EP4 signaling inhibitor in treatment Use in cancer and chronic inflammatory diseases.
  • Prostaglandin E2 is one of the main products produced by the action of cyclooxygenase (COX) on arachidonic acid, and it is known to participate in many physiological and pathophysiological reactions.
  • PGE2-mediated intracellular signaling depends on its binding to one or more specific prostaglandin E receptors (EP1-4) on target cells, which are coupled to different G proteins.
  • EP4 is expressed in a variety of tissues and cells, including immune, bone and joint, cardiovascular, gastrointestinal and respiratory systems, and cancer cells.
  • EP4 coupled with G ⁇ can activate adenylate cyclase (AC) and catalyze the formation of the second messenger cAMP.
  • AC adenylate cyclase
  • cAMP protein kinase A
  • PKA protein kinase A
  • EP4 also stimulates the non-classical pathway phosphatidylinositol 3 kinase (PI3K)/protein kinase B (PKB, also known as Akt) to promote cell survival, and activates extracellular regulatory kinase (ERK) to promote migration and proliferation.
  • PI3K phosphatidylinositol 3 kinase
  • PBB protein kinase B
  • Akt extracellular regulatory kinase
  • EP4 antagonists More and more preclinical data support the potential therapeutic value of prostaglandin E receptor 4 (EP4) antagonists in several indications. Most scientific evidence indicates that selective EP4 antagonists may be an effective inflammatory pain relief drug, and its intestinal tolerance is better than NSAIDs and COX-2 inhibitors. This is the current standard drug. Importantly, EP4 antagonists can achieve safer cardiovascular safety because they do not directly interfere with the biosynthesis of prostaglandin E (PGE2) and other prostaglandins (such as prostacyclin and thromboxane). EP4 receptor antagonists may also have therapeutic applications in the treatment of migraine, because EP4 receptors are involved in PGE2-mediated cerebral vasodilation, which is an important factor in migraine.
  • PGE2 prostaglandin E
  • other prostaglandins such as prostacyclin and thromboxane
  • EP4 receptors play an important role in many neurodegenerative diseases, such as multiple sclerosis and Alzheimer's disease, in which PGE2 is involved.
  • the PGE2/EP4 signaling pathway is related to the occurrence of colorectal cancer, breast cancer, lung cancer, prostate cancer, ovarian cancer, bladder cancer, liver cancer, etc.
  • the activation of this cascade signal, elevated tumor PGE2 levels or EP4 overexpression can inactivate host anti-tumor immune cells, enhance cancer cell proliferation, migration and metastasis, and promote tumor-related angiogenesis, thereby promoting tumor progression.
  • EP4 knockout mice delayed tumorigenesis compared to wild-type mice in the context of APCmin mutation, indicating their tumor promoting activity.
  • Selective EP4 receptor antagonists can inhibit PGE2-induced cancer cell proliferation in vitro, and can slow down tumor progression and metastasis in various preclinical tumor models.
  • the selective EP4 receptor antagonist also blocks the induction of suppressor cells derived from bone marrow, restores the activity of natural killer cells, and enhances the production of pro-inflammatory cytokines (TNF- ⁇ ) and IL-12 by bone marrow cells and Th1 cells.
  • TNF- ⁇ pro-inflammatory cytokines
  • IL-12 pro-inflammatory cytokines
  • E7046 (ie (S)-4-(1-(3-(difluoromethyl)-1-methyl-5-(3-(trifluoromethyl)phenoxy)-1H-pyrazole-4- Carboxamido) ethyl) benzoic acid) is an E-type prostate receptor 4 antagonist developed by Eisai. It changes the tumor microenvironment by acting on the EP4 receptor to promote the immune function of the body. E7046 has demonstrated strong anti-cancer activity and immune benefits in preclinical in vitro and in vivo tests. Combined (such as radiotherapy, immune checkpoint inhibitors) medication can significantly inhibit the growth of a variety of tumors, combined with radiotherapy and chemotherapy. Clinical Ib In progress (WO2015179615A1 and Diana I. Albu et al., "Oncoimmunology", 2017, 6(8): e1338239).
  • MF-766 i.e. 4-(1-(1-(4-(trifluoromethyl)benzyl)-1H-indole-7-carboxamido)cyclopropyl)benzoic acid
  • Merck EP4 receptor antagonist for the treatment of acute and chronic pain, osteoarthritis, rheumatoid arthritis and cancer
  • EP4 receptor antagonists also include, for example, WO2017066633A1, WO2017041323A1, WO2018084230A1 and so on. Due to the huge market demand, it is still necessary to continue to develop highly active and selective EP4 receptor antagonists.
  • the purpose of the present invention is to provide a class of compounds that inhibit PGE2/EP4 signal transduction with high activity and high selectivity.
  • the inventors have repeatedly conducted serious studies and surprisingly found a novel compound containing The compound of formula (I), thus completing the present invention.
  • the present invention relates to compounds of the following general formula (I):
  • M 1 , M 2 , M 3 , M 5 , M 6 and M 7 are each independently a N atom or CR 4 ;
  • M 4 is selected from N atom and C atom
  • Ring A is selected from C 6-10 aryl, C 3-6 cycloalkyl, 5-10 membered heteroaryl and 3-6 membered heterocyclic group;
  • Y is selected from a bond, C 1-4 alkylene, -CR 5 R 6 -, -O-, -OC 1-4 alkylene-, -NR 9 C 1-4 alkylene- and -NR 9 -, the C 1-4 alkylene is optionally selected from H atom, D atom, halogen, hydroxyl, cyano, amino, nitro, C 1-4 alkyl, C 1-4 alkoxy, halogen Substituted C 1-4 alkyl, halogenated C 1-4 alkoxy, hydroxy C 1-4 alkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclic group, C 6-10 aryl and 5 One or more substituents in the -10 membered heteroaryl group;
  • R 1 and R 2 identical or different and are each independently selected from H atom, D atom, a cyano group, C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, 3-6 Membered heterocyclic group, -C(O)OR 4 , C(O)NR 7 R 8 , -COR 4 , -S(O) m R 4 , -NR 7 R 8 , C 6-10 aryl and 5-
  • a 10-membered heteroaryl group, the C 1-4 alkyl group and C 1-4 alkoxy group are each independently optionally selected from H atom, D atom, halogen, hydroxyl, cyano, amino, nitro, C 1 -4 alkyl, C 1-4 alkoxy, halogenated C 1-4 alkyl, halogenated C 1-4 alkoxy, hydroxy C 1-4 alkyl, C 3-6 cycloalkyl, 3- 6-membered heterocyclic group, -C(O)OR 4 ,
  • R 3 are the same or different, and are each independently selected from hydrogen H atom, D atom, halogen, hydroxyl, cyano, amino, nitro, C 1-4 alkyl, C 1-4 alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclyl, C 6-10 aryl, 5-10 membered heteroaryl, -C(O)R 4 , -C(O)OR 4 , -OC(O)NR 7 R 8 , -NR 7 C(O)OR 4 , -S(O) m R 4 , -NR 7 R 8 and -C(O)NR 7 R 8 , wherein the C 1-4 alkyl group, C 1 -4 alkoxy, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, C 6-10 aryl and 5-10 membered heteroaryl are each independently optionally selected from C 1-4 alkyl One of hydroxy C 1-4 alkyl, C 1-4 alkoxy, halogenated C
  • R 4 is selected from H atom, D atom, halogen, hydroxyl, amino, C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl, 3-6 membered heterocyclic group, C 6- 10 aryl and 5-10 membered heteroaryl groups, wherein the C 1-4 alkyl group, C 1-4 alkoxy group, C 3-6 cycloalkyl group, 3-6 membered heterocyclic group, C 6-10 Aryl and 5-10 membered heteroaryl are each independently optionally selected from C 1-4 alkyl, hydroxy C 1-4 alkyl, C 1-4 alkoxy, halogenated C 1-4 alkyl, Substituted by one or more substituents among halogenated C 1-4 alkoxy, halogen, hydroxyl, cyano, amino and nitro groups;
  • R 5 and R 6 are each independently selected from H atom, D atom, C 1-4 alkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclic group, C 6-10 aryl and 5-10 membered Heteroaryl, wherein the C 1-4 alkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclyl, C 6-10 aryl and 5-10 membered heteroaryl are each independently optionally Selected from C 1-4 alkyl, hydroxy C 1-4 alkyl, C 1-4 alkoxy, halogenated C 1-4 alkyl, halogenated C 1-4 alkoxy, halogen, hydroxyl, cyano , Amino and nitro substituted by one or more substituents; or R 5 and R 6 together with the atoms to which they are attached form a C 3-6 cycloalkyl or 3-6 membered heterocyclic group, wherein the C 3 -6 cycloalkyl or 3-6 membered heterocyclic group are
  • R 7 and R 8 are each independently selected from H atom, D atom, C 1-4 alkyl, halogenated C 1-4 alkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclic group, C 6 -10 aryl and 5-10 membered heteroaryl, wherein the C 1-4 alkyl, halogenated C 1-4 alkyl, C 3-6 cycloalkyl, 3-6 membered heterocyclic group, C 6 -10 aryl and 5-10 membered heteroaryl are each independently optionally selected from C 1-4 alkyl, hydroxy C 1-4 alkyl, C 1-4 alkoxy, halogenated C 1-4 alkane Substituted by one or more substituents in the group, halogenated C 1-4 alkoxy, halogen, hydroxy, cyano, amino and nitro;
  • R 9 is selected from H atom, D atom and C 1-4 alkyl group, wherein the C 1-4 alkyl group is optionally selected from hydroxy C 1-4 alkyl group, C 1-4 alkoxy group, halogenated C 1-4 alkyl, halogenated C 1-4 alkoxy, halogen, hydroxy, cyano, amino and nitro substituted by one or more substituents;
  • n 0, 1 or 2;
  • n 0, 1, 2, 3, or 4.
  • the compound represented by general formula (I) is a compound represented by general formula (II):
  • M 1 , M 3 , M 5 and M 6 are each independently CH or C-halogen
  • M 2 is CR 4 or N atom
  • M 7 is N atom
  • M 4 is C atom
  • ring A, R 1 , R 2 , R 3 , R 4 , X, Y and n are as defined in general formula (I).
  • Y is a C 1-4 alkylene group, preferably a methylene group.
  • R 1 and R 2 are the same or different, and are each independently selected from H atom, D atom and C 1-4 alkyl; or R 1 and R 2 together with the atoms to which they are attached form a C 3-6 cycloalkyl group, preferably a cyclopropyl group.
  • the compound represented by general formula (I) is a compound represented by general formula (III):
  • R 1 and R 2 are the same or different, and are each independently selected from H atom, D atom and C 1-4 alkyl; or R 1 and R 2 together with the atoms to which they are connected form a C 3-6 cycloalkyl, preferably Cyclopropyl
  • M 2 is N atom or CR 4 ;
  • R 4 is selected from H atom, D atom and halogen
  • X, ring A, R 3 and n are as defined in the general formula (I).
  • ring A is selected from phenyl, pyridyl, quinolinyl, benzofuranyl, morpholinyl, pyrazole, cyclopropyl , Isoxazole, benzoxazole and benzothiazole.
  • R 3 is the same or different, and each is independently selected from H atom, D atom, halogen, C 1-4 alkyl, fluoro C 1-4 alkyl, phenyl, hydroxy C 1-4 alkyl substituted phenyl, morpholinyl, pyridyl, pyrazolyl, C 1-4 alkyl substituted pyrazolyl, hydroxy C 1-4 Alkyl substituted pyrazolyl, cyclopropyl, isoxazolyl and piperidinyl.
  • Typical compounds of the present invention include but are not limited to:
  • the compounds of the present invention can be prepared by various methods known for preparing such compounds, for example, as shown in the following reaction scheme. Unless otherwise stated in the reaction scheme and subsequent discussion, M 1 to M 7 , ring A, X, Y, R 1 , R 2 , R 3 and n are as defined in the general formula (I).
  • Z Cl, Br or I
  • R is C 1-4 alkyl, preferably methyl.
  • the compound of general formula (I-B) can be obtained by hydrolyzing the compound of general formula (I-A) in solution under alkaline conditions.
  • the hydrolysis reaction can be carried out under conventional conditions: under a typical condition, the reaction is carried out under alkaline conditions, such as in the presence of sodium hydroxide, potassium hydroxide or lithium hydroxide.
  • the selected solvents include methanol, ethanol, propanol, butanol, 2-methoxyethane, 1,4-dioxane, ethylene glycol, tetrahydrofuran, and 1,2-dimethoxyethane.
  • the reaction temperature can be 0-100°C, usually 20°C to 60°C, and the reaction time ranges from 60 minutes to 10 hours.
  • the compound of general formula (IB) and the compound of general formula (IC) undergo a condensation reaction in an inert solvent in the presence of a condensing agent to obtain a compound of general formula (ID).
  • the condensing agent is selected from 2-(7- Benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Amine (EDCI), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-dicyclohexylcarbodiimide, N,N'-diisopropyl Carbodiimide, O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroborate, 1-hydroxybenzotriazole, 1-hydroxy-7-couple Azobenzotriazo
  • the inert solvent is selected from dimethylformamide, dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, tetrahydrofuran, dimethoxyethane or a mixture of these solvents.
  • the reaction temperature can be 0-50°C, and the reaction time can be 10 hours to 24 hours.
  • the compound of general formula (I) can be obtained by hydrolyzing the compound of general formula (I-D) in solution under alkaline conditions.
  • the reaction conditions are the same as in step 2.
  • Z Cl, Br or I
  • R is C 1-4 alkyl, preferably methyl.
  • the compound of general formula (I-1) reacts with the compound of general formula (I-2) in the presence of a base in an inert solvent to obtain the compound of general formula (I-A).
  • the base is selected from potassium tert-butoxide, sodium tert-butoxide, lithium tert-butoxide, sodium hydrogen, potassium hydrogen, sodium ethoxide, potassium ethoxide;
  • the inert solvent is selected from dimethylformamide, dimethyl Acetamide, N-methylpyrrolidone, dimethyl sulfoxide, tetrahydrofuran, dimethoxyethane or a mixture of these solvents.
  • the reaction temperature can be 0-100°C, usually 20°C to 60°C, and the reaction time ranges from 60 minutes to 10 hours.
  • Z Cl, Br or I
  • R is C 1-4 alkyl, preferably methyl.
  • the compound of general formula (I-1) reacts with the compound of general formula (I-3) in the presence of a base in an inert solvent to obtain the compound of general formula (I-4).
  • the reaction conditions are the same as in step 1A.
  • the compound of general formula (I-4) reacts with carbon oxide in methanol or ethanol in the presence of a palladium catalyst, ligand and organic base to obtain a compound of general formula (I-A).
  • the palladium catalyst is selected from the group consisting of palladium acetate, palladium(II) acetylacetonate, palladium(II) propionate, tetrakis(triphenylphosphine)palladium, [1,2-bis(dicyclohexylphosphino)ethane] chloride Palladium(II) and bis(dicyclohexylphosphino)palladium(0)
  • the ligand is selected from triphenylphosphine, 1,2-bis(diphenylphosphino)ethane, di-tert-butyl neopentylphosphine , Bis(2-diphenylphosphinoethyl)phenylphosphine, 1,
  • the present invention also relates to a pharmaceutical composition, which comprises 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.
  • a pharmaceutical composition which comprises 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 present invention also relates to the use of the compound represented by the general formula (I) or the pharmaceutical composition containing the same in the preparation of a medicine for inhibiting PGE2/EP4 signal transduction.
  • the present invention also relates to the use of the compound represented by the general formula (I) or the pharmaceutical composition containing the same in the preparation of a medicine for the treatment of cancer.
  • the cancer is selected from breast cancer, cervical cancer, colorectal cancer, and intrauterine cancer.
  • the present invention also relates to a compound represented by general formula (I) or a pharmaceutical composition containing it for preparing and treating acute or chronic pain, migraine, osteoarthritis, rheumatoid arthritis, gout, bursitis, ankylosing spine Inflammation, primary dysmenorrhea, cancer or arteriosclerosis drugs.
  • a compound represented by general formula (I) or a pharmaceutical composition containing it for preparing and treating acute or chronic pain, migraine, osteoarthritis, rheumatoid arthritis, gout, bursitis, ankylosing spine Inflammation, primary dysmenorrhea, cancer or arteriosclerosis drugs.
  • the present invention also relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, which is used as a medicine.
  • the present invention also relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, which is used to inhibit PGE2/EP4 signaling.
  • the present invention also relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or A pharmaceutically acceptable salt or a pharmaceutical composition containing the same, which is used for the treatment of cancer.
  • the cancer is selected from breast cancer, cervical cancer, colorectal cancer, endometrial cancer, glioblastoma, Head and neck cancer, kidney cancer, liver cancer, lung cancer, medulloblastoma, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer and urethral cancer.
  • the present invention also relates to the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing it, for the treatment of acute or chronic pain, migraine, osteoarthritis, rheumatoid arthritis, gout, bursitis, ankylosing spondylitis, and primary sexual dysmenorrhea, cancer or arteriosclerosis.
  • the compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, diastereomer, or mixture thereof or A pharmaceutically acceptable salt, or a pharmaceutical composition containing it, for the treatment of acute or chronic pain, migraine, osteoarthritis, rheumatoid arthritis, gout, bursitis, ankylosing spondylitis, and primary sexual dysmenorrhea, cancer or arteriosclerosis.
  • a method for inhibiting PGE2/EP4 signal transduction comprising administering a therapeutically effective amount of a compound represented by general formula (I) or its tautomers, mesosomes, racemates, enantiomers to a desired patient Conformers, diastereomers or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing them.
  • a compound represented by general formula (I) or its tautomers, mesosomes, racemates, enantiomers to a desired patient Conformers, diastereomers or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing them.
  • a method for the treatment of cancer comprising administering to a desired patient a therapeutically effective amount of a compound represented by the general formula (I) or its tautomer, meso, racemate, enantiomer, non- Enantiomers or mixtures thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions containing them.
  • the cancer is selected from breast cancer, cervical cancer, colorectal cancer, endometrial cancer, Glioblastoma, head and neck cancer, kidney cancer, liver cancer, lung cancer, medulloblastoma, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer, and urethral cancer.
  • a method for the treatment of acute or chronic pain, migraine, osteoarthritis, rheumatoid arthritis, gout, bursitis, ankylosing spondylitis, primary dysmenorrhea, cancer or arteriosclerosis including administration to patients in need
  • the compounds and antibodies according to the present invention can be administered orally, sublingually, intraperitoneally, 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, and nasal Intra or intraocular administration form and rectal administration form.
  • the main active ingredient is mixed with pharmaceutical carriers such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic and the like.
  • pharmaceutical carriers such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic and the like.
  • the 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.
  • Preparations in the form of syrups or tinctures may contain the active ingredient together with sweetening agents, preservatives, and flavoring agents and appropriate coloring agents.
  • the water-dispersible powders or granules may contain active ingredients mixed with dispersing agents, wetting agents or suspending agents and with flavoring or sweetening agents.
  • Suppositories are used for rectal administration and are prepared with adhesives that melt at rectal temperature, for example, cocoa butter or polyethylene glycol.
  • Aqueous suspension, isotonic saline solution or sterile and injectable solution which contains pharmacologically compatible dispersing agent and/or wetting agent for parenteral, intranasal or intraocular Apply.
  • the active ingredient (possibly with one or more additive carriers) can also be formulated as microcapsules.
  • the compound of the present invention can be administered at a dose 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 1000 mg, even more advantageously between 2.5 mg and 200 mg. It may be necessary to administer doses outside of these ranges, and those skilled in the art will be aware of this themselves.
  • the pharmaceutical composition may also be formulated for external administration. It can be introduced into the usual forms of the application type (ie, in particular lotions, foams, gels, dispersions, sprays), which have excipients, in particular It can penetrate the skin to improve the properties and accessibility of the active ingredients.
  • these compositions usually further comprise a physiologically acceptable medium, which usually comprises water or a solvent, for example, alcohol, ether or 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 filter.
  • stereoisomers refer to geometric isomers (or configuration isomers) or optical isomers.
  • “Geometric isomers” are caused by substituents at different positions on the double bond, which can then have the Z or E configuration, also called cis or trans.
  • Optical isomers are particularly caused by substituents at different steric positions on carbon atoms, said carbon atoms containing four different substituents. This carbon atom constitutes the chiral center or asymmetric center.
  • Optical isomers include diastereomers and enantiomers.
  • Optical isomers that are non-superimposable mirror images of each other are called “enantiomers”.
  • Optical isomers that are not superimposable mirror images of each other are called "diastereomers”.
  • racemic mixture A mixture containing equal amounts of two separate enantiomeric forms of opposite chirality is called a "racemic mixture”.
  • tautomer refers to a structural isomer of a compound obtained by proton transfer rearrangement (prototropie), that is, by the migration of hydrogen atoms and the change of the position of the double bond.
  • the different tautomers of the compound are usually mutually convertible and exist in the solution in a balanced manner in proportions, which may vary according to the solvent used, temperature or pH.
  • pharmaceutically acceptable is understood to mean that it is used to prepare a pharmaceutical composition, which is generally safe, non-toxic, biologically or otherwise satisfactory and the combination The drug can be accepted for veterinary and human drug use.
  • the "pharmaceutically acceptable salt” of the compound is understood to refer to the following salt, which is a pharmaceutically acceptable (as defined herein) salt and which possesses the expected pharmacological activity of the parent compound.
  • This salt includes:
  • Acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., or with organic acids such as acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, and ethanesulfonic acid , Fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, hydroxypamoic acid, 2-hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucon Acid, 2-naphthalenesulfonic acid, propionic acid, salicylic acid, succinic acid, dibenzoyl-L-tartaric acid, tartaric acid, p-toluenesulfonic acid, trimethylacetic acid, trifluoroacetic acid and other acid addition salts; with organic acids such as ace
  • alkali metal ions such as Na + , K + or Li +
  • alkaline earth metal ions such as Ca 2+ or Mg 2+
  • aluminum ions or the salt formed when coordinated with an organic base or an inorganic base.
  • 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 means fluorine, bromine, chlorine or iodine atom.
  • C 1-4 alkyl refers to a saturated straight 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, and 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 -and the like.
  • C 1-4 alkoxy refers to -O-(C 1-4 alkyl), wherein the definition of C 1-4 alkyl is as described 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 by one or more halogens, wherein the definitions of C 1-4 alkyl and halogen are as described above.
  • halogenated C 1-4 alkoxy means that C 1-4 alkoxy is substituted by one or more halogens, wherein C 1-4 alkoxy and halogen are as defined above.
  • hydroxy C 1-4 alkyl refers to hydroxy substituted C 1-4 alkyl, wherein alkyl is 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 a heterocyclic group 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)
  • Atomized saturated or partially unsaturated monocyclic hydrocarbon systems representative examples include, but are not limited to, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, dihydroimidazolyl, dihydroimidazolyl, Hydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, etc.
  • C 6-10 aryl refers to a 6 to 10 membered all-carbon monocyclic or fused polycyclic (that is, rings sharing adjacent pairs of carbon atoms) groups with a conjugated ⁇ -electron system, such as phenyl and naphthalene base. Phenyl is more preferred.
  • the aryl ring may be fused to a heteroaryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, non-limiting examples thereof include:
  • heteroaryl refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 10 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen, preferably containing 1 to 2 heteroatoms 5-membered or 6-membered heteroaryl groups, such as quinolinyl, imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrrolyl, tetrazolyl, pyridyl, Pyrimidine, thiadiazole, pyrazinyl, etc.
  • the heteroaryl ring may be fused on an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:
  • hydroxy 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 "-”.
  • heterocyclic group optionally substituted by an alkyl group means that an alkyl group may but does not have to be present.
  • the description includes the case where the heterocyclic group is substituted by an alkyl group and the case where the heterocyclic group is not substituted by 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 independently of each other substituted by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without too much effort. For example, an amino group or a hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (eg, olefinic) bond.
  • “Pharmaceutical composition” means a mixture containing one or more of the compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs and 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 organism, which is beneficial to the absorption of the active ingredient and thus the biological activity.
  • antibody includes all types of immunoglobulins, including IgG, IgM, IgA, IgD, and IgE, or fragments thereof, which can be applied to the medicine used herein.
  • the antibody may be a monoclonal antibody or a polyclonal antibody, and may be a species of any origin, including, for example, mouse, rat, rabbit, horse, or human.
  • Retaining antibody fragments found specifically for proteins or epitopes, such as CTLA4, PDL1 or PD1, which bind to the antibodies used in the present invention, are also included in the scope of the term "antibody”. These fragments can be produced by known techniques.
  • Antibodies can be chimeric or humanized, especially when they are used for therapeutic purposes.
  • CTLA4 antibody or “anti-CTLA4" refers to an antibody or antibody against cytotoxic T-lymphocyte antigen 4 (CTLA4).
  • exemplary antibodies include, but are not limited to, CTLA4 antagonist antibodies or CTLA4 antibodies, such as Ipilimumab (Ipilimumab, Bristol-Myers Squibb) and Tremelimumab (Pfizer).
  • PDL1 antibody or “anti-PDL1” refers to an antibody against programmed death ligand 1 (PDL1).
  • Example antibodies include but are not limited to Atezolizumab (Roche), Devalumab (Durvlumab, AstraZeneca), BMS-936559, CK-301 (Check Point Therapeutics), KN035 (Corning Jereh), BGB-A333 (BeiGene), CS1001 (Cornerstone), HLX20 (Fuhong Henlius), KL-A167 (Kelun Botai Company), F520 (New Times Company), GR1405 (Zhixiang Jintai Company), MSB2311 (Maebs Company).
  • PD1 antibody or “anti-PD1” refers to an antibody against programmed death protein 1 (PD1).
  • Example antibodies include, but are not limited to nivolumab (nivolumab, Bristol-Myers Squibb), labrolizumab (Merck), pembrolizumab (Merck & Co.), Avelumab (Merck/Pfizer), Pidilizumab (Medivation Company), AMP-224, AMP-514 (GSK), Spartalizumab (Novartis), Cemiplimab (trade name Libtayo, Sanofi/Regeneron), Treplimab (Junshi), Sintilizumab (Cinda), Carrelizumab (Hengrui Company), Tirelizumab (BeiGene), GLS-010 (Yuheng Pharmaceutical Company), GB226 (Jiahe Company) ), CS1003 (Cornerstone Company), BAT-1306 (Bai Aota
  • Figure 1 shows the effect of different doses of compound 6 and E7046 on tumor growth in a CT-26 colorectal cancer model.
  • Figure 2 shows the effect of different doses of Compound 6 and E7046 on the body weight of mice in a CT-26 colorectal cancer model.
  • Figure 3 shows the effect of different doses of compound 6 and anti-PD-1 antibody on tumor growth when administered separately or in combination in a CT-26 colorectal cancer model.
  • Figure 4 shows the effect of different doses of Compound 6 and anti-PD-1 antibody on the body weight of mice when administered alone and in combination in a CT-26 colorectal cancer model.
  • Figure 5 shows the effect of different doses of Compound 6 and anti-PD-1 antibody on tumor growth when administered alone and in combination in an EMT-6 breast cancer model.
  • Figure 6 shows the effect of different doses of Compound 6 and anti-PD-1 antibody on the body weight of mice when administered alone and in combination in a breast cancer tumor model.
  • the experimental methods without specific conditions are usually in accordance with conventional conditions or in accordance with the conditions recommended by the raw material or commodity manufacturers.
  • Reagents without specific sources are conventional reagents purchased on the market.
  • 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 determination solvent is deuterated chloroform (CDCl 3 ), deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated methanol (CD 3 OD) or deuterated acetonitrile (CD 3 CN).
  • the internal standard is tetramethylsilane (TMS).
  • s is a singlet
  • bs is a broad singlet
  • d is a doublet
  • t is a triplet
  • q is a quartet
  • m is a multiplet or a large number of peaks
  • dd is a double doublet
  • Liquid Mass Spectrometry SHIMADZU LCMS-2020, Column: 5 ⁇ m x 30x2.1mm S/N: H17-247175, column temperature: 50.0°C, mobile phase: A: water (0.0375% TFA), B: acetonitrile (0.01875% TFA), ionization mode: ESI, polarity: positive.
  • Nuclear magnetic resonance instrument Bruker ARX-500 high resolution mass spectrometer and Bruker ARX-400 high resolution mass spectrometer.
  • MTT detection instrument Thermo Scientific Multiskan GO full-wavelength microplate reader.
  • the thin layer chromatography silica gel plate uses Qingdao GF254 silica gel plate, the size of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm ⁇ 0.2mm, and the size of thin layer chromatography separation and purification products is 0.4mm ⁇ 0.5mm.
  • the reactions are all 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.
  • TLC thin layer chromatography
  • cAMP cyclic adenosine monophosphate
  • PI3K Also stimulates the non-classical pathway phosphatidylinositol 3 kinase
  • ERK Extracellular regulatory kinase
  • NSAIDs Non-steroidal anti-inflammatory drugs
  • TNF- ⁇ Tumor Necrosis Factor ⁇
  • IL-12 Interleukin 12
  • HATU 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine-3-oxide hexafluorophosphate
  • DIPEA Diisopropylethylamine
  • NBS N-bromosuccinimide
  • tBuBrettPhos 2-(di-tert-butylphosphino)-2',4',6'-triisopropyl-3,6-dimethoxy-1,1'-biphenyl
  • Step 2 Preparation of 1-(4-(trifluoromethyl)benzyl)-1H-indole-7-carboxylic acid 1-3
  • Step 3 3-(1-(1-(4-(Trifluoromethyl)benzyl)-1H-indole-7-carboxamido)cyclopropyl)bicyclo[1.1.1]pentan Preparation of methyl alkane-1-carboxylate 1-4
  • Step 4 3-(1-(1-(4-(Trifluoromethyl)benzyl)-1H-indole-7-carboxamido)cyclopropyl)bicyclo[1.1.1]pentan Preparation of alkane-1-carboxylic acid 1
  • Step 3 Preparation of 1-((5-phenylpyridin-2-yl)methyl)-1H-indole-7-carboxylic acid methyl ester 2-4
  • Step 4 Preparation of 1-((5-phenylpyridin-2-yl)methyl)-1H-indole-7-carboxylic acid 2-5
  • Step 5 3-(1-(1-((5-phenylpyridin-2-yl)methyl)-1H-indole-7-carboxamido)cyclopropyl)bicyclic [1.1.1] Preparation of methyl pentane-1-carboxylate 2-6
  • Step 6 3-(1-(1-((5-phenylpyridin-2-yl)methyl)-1H-indole-7-carboxamido)cyclopropyl)bicyclic [1.1.1] Preparation of pentane-1-carboxylic acid 2
  • Step 3-6 3-(1-(1-((6-phenylpyridin-3-yl)methyl)-1H-indole-7-carboxamido)cyclopropyl)bicyclic [1.1.1] Preparation of pentane-1-carboxylic acid 5
  • Step 2-5 3-(1-(5-chloro-1-((5-phenylpyridin-2-yl)methyl)-1H-indole-7-carboxamido)cyclopropyl) Preparation of bicyclo[1.1.1]pentane-1-carboxylic acid 7
  • Step 4 Preparation of 1-((5-morpholinopyridin-2-yl)methyl)-1H-indole-7-carboxylic acid 10-5
  • Step 5 3-(1-(1-((5-morpholinopyridin-2-yl)methyl)-1H-indole-7-carboxamido)cyclopropyl)bicyclic [1.1.1] Preparation of methyl pentane-1-carboxylate 10-6
  • Step 1 Preparation of 1-([2,3'-bipyridine]-6'-ylmethyl)-1H-indole-7-carboxylic acid 11-1
  • Step 2 3-(1-(1-([2,3'-Bipyridyl]-6'-ylmethyl)-1H-indole-7-carboxamido)cyclopropyl)bicyclic [1.1.1] Preparation of methyl pentane-1-carboxylate 11-2
  • Step 3 3-(1-(1-([2,3'-bipyridyl]-6'-ylmethyl)-1H-indole-7-carboxamido)cyclopropyl)bicyclic [1.1.1] Preparation of pentane-1-carboxylic acid 11
  • step 1 compound 11-0 was replaced by pyridin-3-ylboronic acid to obtain compound 12 as a pale yellow solid.
  • step 1 compound 11-0 was replaced by pyridin-4-ylboronic acid to obtain compound 13, as a light yellow solid.
  • step 1 (1-methyl-1H-pyrazol-4-yl)boronic acid was substituted for compound 11-0 to obtain compound 14 as a white solid.
  • step 1 (1H-pyrazol-4-yl)boronic acid was substituted for compound 11-0 to obtain compound 15, as a pale yellow solid.
  • step 1 3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 1H-pyrazol-1-yl)propan-1-ol was substituted for compound 11-0 to obtain compound 16, a white solid.
  • step 1 piperidin-3-ol was used to replace compound 11-0.
  • Cs 2 CO 3 (787mg, 2.42mmol)
  • 2-(dimethylamino) was added.
  • Acetic acid 25mg, 242umol
  • cuprous iodide 70.0mg, 367umol
  • step 1 cyclopropylboronic acid was substituted for compound 11-0 to obtain compound 18 as a white solid.
  • step 1 (2-(hydroxymethyl)phenyl)boronic acid was substituted for compound 11-0 to obtain compound 19 as a white solid.
  • Step 1 Preparation of 3-(chlorocarbonyl)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester 20-1
  • Step 5 3-(1-(1-(benzofuran-2-ylmethyl)-1H-indole-7-carboxamido)ethyl)bicyclo[1.1.1]pentan Preparation of alkane-1-carboxylic acid 20
  • Step 6 (R)-3-(1-(1-(benzofuran-2-ylmethyl)-1H-indole-7-carboxamido)ethyl)bicyclo[1.1.1] Pentane-1-carboxylic acid 20-5 and (S)-3-(1-(1-(benzofuran-2-ylmethyl)-1H-indole-7-carboxamido)ethyl)bis Preparation of cyclo[1.1.1]pentane-1-carboxylic acid 20-6
  • reaction solution was poured into water, extracted with ethyl acetate (200mL x 3), the organic phase was washed with water and saturated brine in turn, dried with anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain the crude product, which was passed through a silica gel column [ Eluent: petroleum ether-ethyl acetate (20:1-5:1)] to obtain compound 21-1 (2.1 g, yield: 51.8%) as a yellow solid.
  • Step 4 3-(1-(1-(1-(benzofuran-2-ylmethyl)-1H-pyrrolo[3,2-c]pyridine-7-carboxamido) ring Synthesis of propyl)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester 25-4
  • benzofuran-2-carboxylic acid methyl ester 30-0 (1.9 g, 10.8 mmol) and tetrahydrofuran (50 mL) were added, stirred under nitrogen protection, and cooled to 0° C. in an ice bath.
  • Lithium tetrahydroaluminum-d4 (2.2g, 54.10mmol) was slowly added to it in batches, keeping the temperature at 0°C. After the addition is complete, continue to stir at 0°C for 1 hour.
  • Step 2 Synthesis of 3-(1-aminocyclopropyl-2,2,3,3-d4)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester
  • the aqueous phase was basified to pH ⁇ 12 with 10M aqueous sodium hydroxide solution at 0°C and extracted with ethyl acetate (200 mL ⁇ 2).
  • compound 6-2 (251mg, 1.17mmol) and N,N-dimethylformamide (10mL) were added, cooled to 0°C in an ice bath with stirring, and then potassium tert-butoxide (171mg, 1.52mmol) and 7-bromo-5-fluoro-1H-indole 32-0 (295mg, 1.40mmol). After the addition is complete, stir at room temperature for 1 hour.
  • Test example 1 MCF7 cAMP measurement
  • Inhibition% 100-100 ⁇ (high control-analysis data)/(high control-low control)
  • Test example 2 Clearance test in liver cells
  • Hepatocyte type mouse
  • b Transfer a vial of cryopreserved liver cells from storage, ensuring that the vial is kept at a low temperature until the thawing process occurs. Thaw the cells by placing the vial in a 37°C water bath and gently shaking the vial for 2 minutes. After the thawing is complete, spray the vial with 70% ethanol, and transfer the vial to a biological safety cabinet.
  • the in vitro half-life (t1/2) is determined by the slope value k:
  • N number of hepatocytes per well (0.1 ⁇ 10 6 cells)
  • Test Example 3 Inhibitory effect of compound 6 on CT-26 colorectal tumors in mice
  • mice 6-week-old female BALB/c mice, each implanted subcutaneously with 1 ⁇ 10 6 CT26 cancer cells (National Experimental Cell Resource Sharing Service Platform, resource number 3111CCCC), when the tumor reaches approximately At 60-80mm 3 , mice bearing tumors were randomly assigned to the vehicle or treatment group.
  • Compound 6 was administered orally (po) as a suspension in 0.5% MC at an amount of 10, 30, or 150 mg/kg, once a day.
  • the mice in the vehicle group were given oral distilled water repeatedly every day, and the administration time was the same as that of compound 6.
  • the tumor volume and body weight were recorded 2-3 times a week. The results are shown in Figure 1 and Figure 2.
  • FIG. 1 shows that compound 6 and E7046 have no significant effect on the body weight of mice.
  • mice 6-week-old female BALB/c mice were implanted subcutaneously with 1 ⁇ 10 6 CT26 cancer cells. When the tumor reached about 60-80mm 3 , the mice bearing the tumor were randomly assigned To the vehicle or treatment group.
  • Compound 6 was administered orally (po) as a suspension in 0.5% MC at an amount of 30 or 150 mg/kg when administered alone or in combination, once a day.
  • anti-PD-1 antibody Intraperitoneally at a dose of 5 mg/kg, once every 4 days, for a total of 3 times (Q4D ⁇ 3).
  • the mice in the vehicle group and PD-1 administered alone were given oral distilled water repeatedly every day, and the administration time was the same as that of compound 6.
  • the tumor volume and body weight were recorded 2-3 times a week. The results are shown in Figure 3 and Figure 4.
  • Figure 4 shows that Compound 6 has no significant effect on the body weight of mice when administered alone and in combination with anti-PD1 antibody.
  • Test Example 5 Compound 6 and anti-PD-1 antibody alone and in combination can affect EM-6 mammary glands in mice Tumor suppression
  • mice 6-week-old female BALB/c mice were implanted subcutaneously with 5 ⁇ 10 5 EMT-6 cancer cells. When the tumor reached about 60-80 mm 3 , the mice bearing the tumor were Randomly assigned to the vehicle or treatment group.
  • Compound 6 was administered orally (po) in a suspension of 30 mg/kg in 0.5% MC when administered alone or in combination, once a day.
  • an anti-PD-1 antibody Invivomab anti-mouse PD-1 (CD279), Abcam
  • the mice in the vehicle group and PD-1 administered alone were given oral distilled water repeatedly every day, and the administration time was the same as that of compound 6.
  • the tumor volume and body weight were recorded 2-3 times a week. The results are shown in Figure 5 and Figure 6.
  • Figure 6 shows that Compound 6 has no significant effect on the body weight of mice when administered alone and in combination with anti-PD1 antibody.

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Abstract

一种通式(I)所示的化合物、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PGE2/EP4信号传导抑制剂的用途以及用于治疗癌症、急性或慢性疼痛、偏头痛、骨关节炎、类风湿性关节炎、痛风、粘液囊炎、强直性脊柱炎、原发性痛经、癌症或动脉硬化症的用途。

Description

抑制PGE2/EP4信号传导的化合物、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PGE2/EP4信号传导抑制剂在治疗癌症和慢性炎症性疾病中的用途。
背景技术
前列腺素E2(PGE2)是环加氧酶(COX)对花生四烯酸的作用产生的主要产物之一，并且众所周知其参与许多生理和病理生理反应。PGE2介导的细胞内信号传导取决于其与靶细胞上的一种或多种特异性前列腺素E受体(EP1-4)的结合，这些受体与不同的G蛋白偶联。EP4在多种组织和细胞中表达，包括免疫、骨关节、心血管、胃肠和呼吸系统以及癌细胞。EP4与Gα偶联可激活腺苷酸环化酶(AC)并催化第二信使cAMP的形成。cAMP的主要作用是结合并激活蛋白激酶A(PKA)，其反过来磷酸化细胞中的靶蛋白。此外，EP4还刺激非经典途径磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(PKB，也称为Akt)从而促进细胞存活，并激活细胞外调节激酶(ERK)从而促进迁移和增殖。
越来越多的临床前数据支持前列腺素E受体4(EP4)拮抗剂在几种适应症中的潜在治疗价值。大多数科学证据表明，选择性EP4拮抗剂可能是一种有效的炎症性疼痛缓解药物，其肠道耐受性优于NSAIDs和COX-2抑制剂，这是目前的标准药物。重要的是，EP4拮抗剂可以达成更安全的心血管安全性，因为它们不直接干扰前列腺素E(PGE2)和其他前列腺素(例如前列环素和血栓素)的生物合成。EP4受体拮抗剂也可以在治疗偏头痛中具有治疗应用，因为EP4受体参与PGE2介导的脑血管扩张是偏头痛的重要因素。
COX2在各种肿瘤类型中的过表达导致PGE2水平增加，表明在癌症治疗期间用选择性EP4拮抗剂阻断PGE2信号传导可能对肿瘤学环境中的患者有益。据报道，EP4受体在许多神经退行性疾病中发挥重要作用，例如多发性硬化和阿尔茨海默病，其中涉及PGE2。
研究表明，PGE2/EP4信号通路与结直肠癌、乳腺癌、肺癌、前列腺癌、卵巢癌、膀胱癌、肝癌等的发生有关。这种级联信号的激活，肿瘤PGE2水平升高或EP4过表达能够通过失活宿主抗肿瘤免疫细胞，增强癌细胞增殖、迁移和转移，促进肿瘤相关的血管生成等方式，从而促进肿瘤进展。相比之下，EP4敲除小鼠在APCmin突变的背景下与野生型小鼠相比延迟了肿瘤发生，表明其肿瘤促进活性。
选择性EP4受体拮抗剂在体外能够抑制PGE2诱导的癌细胞增殖，并且在各种临床前肿瘤模型中能够减缓肿瘤进展和转移。选择性EP4受体拮抗剂还阻断骨髓来源的抑制细胞的诱导，恢复自然杀伤细胞活性，并增强骨髓细胞和Th1细胞的促炎细胞因子(TNF-α)和IL-12产生。这些数据表明抑制PGE2/EP4信号传导可能在癌症和其他慢性炎症性疾病中具有治疗价值。因此，生成能够阻断PGE2/EP4信号传导途径的新化合物来填补未满足的医学需求是非常重要的。
E7046(即(S)-4-(1-(3-(二氟甲基)-1-甲基-5-(3-(三氟甲基)苯氧基)-1H-吡唑-4-甲酰胺基)乙基)苯甲酸)是由卫材公司开发的一种E型前列腺受体4拮抗剂，通过作用于EP4受体改变肿瘤微环境从而促进机体免疫作用。E7046在临床前体外和体内试验中已经验证了强的抗癌活性和免疫获益，联合(如放疗、免疫检查点抑制剂)用药能够显著抑制多种肿瘤的生长，与放化疗联合的临床Ⅰb正在进行中(WO2015179615A1和Diana I.Albu等人，《Oncoimmunology》，2017，6(8):e1338239)。
Figure PCTCN2020072487-appb-000001
MF-766(即4-(1-(1-(4-(三氟甲基)苄基)-1H-吲哚-7-甲酰胺基)环丙基)苯甲酸)是由默克公司开发的EP4受体拮抗剂，用于治疗急性和慢性疼痛、骨关节炎、类风湿性关节炎和癌症(WO2008104055A1)。
Figure PCTCN2020072487-appb-000002
目前已有的EP4受体拮抗剂专利申请还包括，如WO2017066633A1、WO2017041323A1、WO2018084230A1等等。由于巨大的市场需求，所以仍有必要继续研发高活性和高选择性的EP4受体拮抗剂。
发明内容
本发明的目的在于提供一类高活性和高选择性的抑制PGE2/EP4信号传导的化合物，发明人为了达到该目的反复进行了认真研究，令人惊奇地发现一种新颖的含有
Figure PCTCN2020072487-appb-000003
的通式(I)化合物，从而完成了本发明。
本发明涉及下面通式(I)的化合物：
Figure PCTCN2020072487-appb-000004
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：
M 1、M 2、M 3、M 5、M 6和M 7各自独立地为N原子或C-R 4；
M 4选自N原子和C原子；
环A选自C 6-10芳基、C 3-6环烷基、5-10元杂芳基和3-6元杂环基；
X为
Figure PCTCN2020072487-appb-000005
Y选自一个键、C 1-4亚烷基、-CR 5R 6-、-O-、-OC 1-4亚烷基-、-NR 9C 1-4亚烷基-和-NR 9-，所述C 1-4亚烷基任选被选自H原子、D原子、卤素、羟基、氰基、氨基、硝基、C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、羟基C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基中的一个或多个取代基所取代；
R 1和R 2相同或不同，且各自独立地选自H原子、D原子、氰基、C 1-4烷基、C 1-4烷氧基、C 3-6环烷基、3-6元杂环基、-C(O)OR 4、C(O)NR 7R 8、-COR 4、-S(O) mR 4、-NR 7R 8、C 6-10芳基和5-10元杂芳基，所述C 1-4烷基和C 1-4烷氧基各自独立地任选被选自H原子、D原子、卤素、羟基、氰基、氨基、硝基、C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、羟基C 1-4烷基、C 3-6环烷基、3-6元杂环基、-C(O)OR 4、C(O)NR 7R 8、-COR 4、-NR 4C(O)NR 7R 8、-OC(O)NR 7R 8、-NR 7C(O)OR 4、-S(O) mR 4、-NR 7R 8、C 6-10芳基和5-10元杂芳基中的一个或多个取代基所取代；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，其中所述C 3-6环烷基和3-6元杂环基各自独立地任选被选自D原子、素、羟基、氰基、氨基、硝基、-C(O)OR 4、C(O)NR 7R 8、-COR 4、-NR 4C(O)NR 7R 8、-OC(O)NR 7R 8、-NR 7C(O)OR 4、-S(O) mR 4、-NR 7R 8、C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、羟基C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基中的一个或多个取代基所取代；
R 3相同或不同，且各自独立地选自氢H原子、D原子、卤素、羟基、氰基、氨基、硝基、C 1-4烷基、C 1-4烷氧基、C 3-6环烷基、3-6元杂环基、C 6-10芳基、5-10元杂芳基、-C(O)R 4、-C(O)OR 4、-OC(O)NR 7R 8、-NR 7C(O)OR 4、-S(O) mR 4、-NR 7R 8和-C(O)NR 7R 8，其中所述C 1-4烷基、C 1-4烷氧基、C 3-6环烷基、3-6元杂环基、C 6-10 芳基和5-10元杂芳基各自独立地任选被选自C 1-4烷基、羟基C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、卤素、羟基、氰基、氨基和硝基中的一个或多个取代基所取代；
R 4选自H原子、D原子、卤素、羟基、氨基、C 1-4烷基、C 1-4烷氧基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基，其中所述C 1-4烷基、C 1-4烷氧基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基各自独立地任选被选自C 1-4烷基、羟基C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、卤素、羟基、氰基、氨基和硝基中的一个或多个取代基所取代；
R 5和R 6各自独立地选自H原子、D原子、C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基,其中所述C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基各自独立地任选被选自C 1-4烷基、羟基C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、卤素、羟基、氰基、氨基和硝基中的一个或多个取代基所取代；或者R 5和R 6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，其中所述C 3-6环烷基或3-6元杂环基各自独立地任选被选自卤素、羟基、氰基、氨基、硝基、C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、羟基C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基中的一个或多个取代基所取代；
R 7和R 8各自独立地选自H原子、D原子、C 1-4烷基、卤代C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基，其中所述C 1-4烷基、卤代C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基各自独立地任选被选自C 1-4烷基、羟基C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、卤素、羟基、氰基、氨基和硝基中的一个或多个取代基所取代；
R 9选自H原子、D原子和C 1-4烷基，其中所述C 1-4烷基任选被选自羟基C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、卤素、羟基、氰基、氨基和硝基中的一个或多个取代基所取代；
m为0、1或2；以及
n为0、1、2、3或4。
在一个优选实施例方案中，通式(I)所示的化合物为通式(II)所示的化合物：
Figure PCTCN2020072487-appb-000006
M 1、M 3、M 5和M 6各自独立地为CH或C-卤素，M 2为C-R 4或N原子，M 7为N原子，M 4为C原子，环A、R 1、R 2、R 3、R 4、X、Y和n如通式(I)所定义。
在另一个优选实施例方案中，在通式(I)所示的化合物中，Y为C 1-4亚烷基，优选亚甲基。
在另一个优选实施例方案中，在通式(I)所示的化合物中，R 1和R 2相同或不同，且各自独立地选自H原子、D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，优选环丙基。
在另一个优选实施例方案中，通式(I)所示的化合物为通式(III)所示的化合物：
Figure PCTCN2020072487-appb-000007
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可药用的盐，
其中：
R 1和R 2相同或不同，且各自独立地选自H原子、D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，优选环丙基；
M 2为N原子或C-R 4；
R 4选自H原子、D原子和卤素；
X、环A、R 3和n如通式(I)中所定义。
在另一个优选实施例方案中，在通式(I)所示的化合物中，环A选自苯基、吡啶基、喹啉基、苯并呋喃基、吗啉基、吡唑、环丙基、异噁唑、苯并噁唑和苯并噻唑。
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相同或不同，且各自独立地选自H原子、D原子、卤素、C 1-4烷基、氟代C 1-4烷基、苯基、羟基C 1-4烷基取代的苯基、吗啉基、吡啶基、吡唑基、C 1-4烷基取代的吡唑基、羟基C 1-4烷基取代的吡唑基、环丙基、异噁唑基和哌啶基。
本发明的典型化合物包括但不限于：
Figure PCTCN2020072487-appb-000008
Figure PCTCN2020072487-appb-000009
Figure PCTCN2020072487-appb-000010
Figure PCTCN2020072487-appb-000011
Figure PCTCN2020072487-appb-000012
Figure PCTCN2020072487-appb-000013
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。
本发明的化合物可以通过制备这类化合物已知的多种方法来制备，例如以下反应方案所示。除非在反应方案和随后的讨论中另有说明，否则M 1至M 7、环A、X、Y、R 1、R 2、R 3和n如通式(I)中所定义。
以下反应方案说明了通式(I)所示的化合物的制备方法。
反应方案1：通式(I)所示的化合物的制备方法
Figure PCTCN2020072487-appb-000014
其中：
Z＝Cl、Br或者I；
R为C 1-4烷基，优选甲基。
步骤1
在这步反应中，通式(I-B)的化合物可以通过通式(I-A)的化合物在碱性条件下溶液中水解获得。水解反应可以在常规条件下进行：在一个典型的条件下，反应在碱性条件，如在有氢氧化钠、氢氧化钾或者氢氧化锂的存在下进行。所选的溶剂包括甲醇、乙醇、丙醇、丁醇、2-甲氧基乙烷、1,4-二噁烷、乙二醇、四氢呋喃和1,2-二甲氧基乙烷。反应温度可以是在0-100℃,通常在20℃到60℃,反应时间从60分钟到10小时。
步骤2
通式(I-B)的化合物和通式(I-C)的化合物在缩合剂存在下在惰性溶剂中发生缩合反应得到通式(I-D)的化合物，优选地，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-乙基-3-(3-二甲基氨基丙基)碳二亚胺(EDCI)、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-羟基苯并三唑、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三唑-1-基-氧基三吡咯烷基磷，最优选2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU)和1-乙基-3-(3-二甲基氨基丙基)碳二亚胺(EDCI)。所述惰性溶剂选自二甲基甲酰胺、二甲基乙酰胺、N-甲基吡咯烷酮、二甲亚砜,四氢呋喃、二甲氧基乙烷或者这些溶剂的混合物。反应温度可以是在0-50℃,反应时间从10小时到24小时。
步骤3
在这步反应中，通式(I)的化合物可以通过通式(I-D)的化合物在碱性条件下溶液中水解获得。反应条件相同于步骤2。
反应方案2：中间体通式(I-A)的化合物的制备方法一
Figure PCTCN2020072487-appb-000015
其中：
Z＝Cl、Br或者I；
R为C 1-4烷基，优选甲基。
在这步反应中，通式(I-1)的化合物同通式(I-2)的化合物在碱的存在下在惰性溶剂中反应得到通式(I-A)的化合物。优选地，所述碱选自叔丁醇钾、叔丁醇钠、叔丁醇锂、钠氢、钾氢、乙醇钠、乙醇钾；所述惰性溶剂选自二甲基甲酰胺、二甲基乙酰胺、N-甲基吡咯烷酮、二甲亚砜、四氢呋喃、二甲氧基乙烷或者这些溶剂的混合物。反应温度可以是在0-100℃,通常在20℃到60℃,反应时间从60分钟到10小时。
反应方案3：中间体通式(I-A)的化合物的制备方法二
Figure PCTCN2020072487-appb-000016
其中：
Z＝Cl、Br或者I；
R为C 1-4烷基，优选甲基。
步骤1
在这步反应中，通式(I-1)的化合物同通式(I-3)的化合物在碱的存在下在惰性溶剂中反应得到通式(I-4)的化合物。反应条件同于步骤1A。
步骤2
在这步反应中，通式(I-4)的化合物在钯催化剂、配体和有机碱的存在下，在甲醇或乙醇中同一氧化碳反应得到通式(I-A)的化合物。优选地，钯催化剂选自醋酸钯、乙酰丙酮钯(II)、丙酸钯(II)、四(三苯基膦)钯、[1,2-双(二环己基膦基)乙烷]氯化钯(II)和双(二环己基膦基)钯(0)，配体选自三苯膦、1,2-双(二苯基膦基)乙烷、二叔丁基新戊基膦、双(2-二苯基膦乙基)苯基膦、1,2-双(二环己基膦基)乙烷、tBuBrettPhos和((2,4,6-三异丙基)苯基)二环己基膦。反应温度可以是60到120℃,反应时间从24小时到48小时。
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。
本发明还涉及通式(I)所示的化合物或包含其的药物组合物在制备抑制PGE2/EP4信号传导的药物中的用途。
本发明还涉及通式(I)所示的化合物或包含其的药物组合物在制备治疗癌症的药物中的用途，优选地，所述癌症选自由乳腺癌、宫颈癌、结肠直肠癌、子宫内膜癌、胶质母细胞瘤、头颈癌、肾癌、肝癌、肺癌、髓母细胞瘤、卵巢癌、胰腺癌、前列腺癌、皮肤癌和尿道癌。
本发明还涉及通式(I)所示的化合物或包含其的药物组合物在制备治疗急性或慢性疼痛、偏头痛、骨关节炎、类风湿性关节炎、痛风、粘液囊炎、强直性脊柱炎、原发性痛经、癌症或动脉硬化症的药物中的用途。
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，其用于抑制PGE2/EP4信号传导。
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，其用于治疗癌症，优选地，所述癌症选自由乳腺癌、宫颈癌、结肠直肠癌、子宫内膜癌、胶质母细胞瘤、头颈癌、肾癌、肝癌、肺癌、髓母细胞瘤、卵巢癌、胰腺癌、前列腺癌、皮肤癌和尿道癌。
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，其用于治疗急性或慢性疼痛、偏头痛、骨关节炎、类风湿性关节炎、痛风、粘液囊炎、强直性脊柱炎、原发性痛经、癌症或动脉硬化症。
一种抑制PGE2/EP4信号传导的方法，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。
一种治疗癌症的方法，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，优选地，所述癌症选自由乳腺癌、宫颈癌、结肠直肠癌、子宫内膜癌、胶质母细胞瘤、头颈癌、肾癌、肝癌、肺癌、髓母细胞瘤、卵巢癌、胰腺癌、前列腺癌、皮肤癌和尿道癌。
一种治疗急性或慢性疼痛、偏头痛、骨关节炎、类风湿性关节炎、痛风、粘液囊炎、强直性脊柱炎、原发性痛经、癌症或动脉硬化症的方法，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。
根据本发明的化合物和抗体可以被口服施用、舌下施用、腹膜内、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、阿拉伯胶等混合。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/天之间的剂量来施用，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1000mg之间，甚至更有利地介于2.5mg和200mg之间。施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 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“羟基C 1-4烷基”指被羟基取代的C 1-4烷基，其中烷基如上所定义。
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系统，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0、1或2)的杂原子的饱和或部分不饱和单环烃系统，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“C 6-10芳基”指具有共轭的π电子体系的6至10元全碳单环或稠合多环(也就是共享毗邻碳原子对的环)基团，例如苯基和萘基。更优选苯基。所述芳基环可以稠合于杂芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为芳基环，其非限制性实例包括：
Figure PCTCN2020072487-appb-000017
术语“5-10元杂芳基”指包含1至4个杂原子、5至10个环原子的杂芳族体系，其中杂原子选自氧、硫和氮，优选含有1至2个杂原子的5元或6元杂芳基，例如喹啉基、咪唑基、呋喃基、噻吩基、噻唑基、吡唑基、噁唑基、异噁唑基、吡咯基、四唑基、吡啶基、嘧啶基、噻二唑、吡嗪基等。所述杂芳基环可以稠合于芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为杂芳基环，其非限制性实例包括：
Figure PCTCN2020072487-appb-000018
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。
“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。
如本文所用的术语“抗体”包括所有类型的免疫球蛋白，包括IgG，IgM，IgA，IgD和IgE，或其片段，其可适用于本文所用的医学。抗体可以是单克隆抗体或多克隆抗体，并且可以是任何来源的物种，包括例如小鼠、大鼠、兔、马或人。保留对蛋白质或表位特异性发现的抗体片段，例如CTLA4、PDL1或PD1，与本发明中使用的抗体结合，也包括在术语“抗体”的范围内。这些片段可以通过已知的技术产生。抗体可以是嵌合的或人源化的，特别是当它们用于治疗目的时。
“CTLA4抗体”或“抗-CTLA4”是指针对细胞毒性T-淋巴细胞抗原4(CTLA4)的抗体或抗体。实示例抗体包括但不限于CTLA4拮抗剂的抗体或CTLA4抗体，如易普利姆玛单抗(Ipilimumab，百时美施贵宝公司)和Tremelimumab(辉瑞公司)。
“PDL1抗体”或“抗PDL1”是指针对程序性死亡配体1(PDL1)的抗体，实例抗体包括但不限于Atezolizumab(罗氏公司)、德瓦鲁单抗(Durvlumab，阿斯利康公司)、BMS-936559、CK-301(Check Point Therapeutics公司)、KN035(康宁杰瑞公司)、BGB-A333(百济神州公司)、CS1001(基石公司)、HLX20(复宏汉霖公司)、KL-A167(科伦博泰公司)、F520(新时代公司)、GR1405(智翔金泰公司)、MSB2311(迈博斯公司)。
“PD1抗体”或“抗-PD1”是指针对程序性死亡蛋白1(PD1)的抗体。实例抗体包括但不限于纳武单抗(nivolumab，百时美施贵宝公司)、labrolizumab(默克公司)、派姆单抗(pembrolizumab，默沙东公司)、Avelumab(默克/辉瑞公司)、Pidilizumab(Medivation公司)、AMP-224、AMP-514(葛兰素史克)、Spartalizumab(诺华公司)、Cemiplimab(商品名Libtayo，赛诺菲/再生元公司)、特瑞普利单 抗(君实公司)、信迪利单抗(信达)、卡瑞利株单抗(恒瑞公司)、替雷利株单抗(百济神州公司)，GLS-010(誉衡药业公司)、GB226(嘉和公司)、CS1003(基石公司)、BAT-1306(白奥泰公司)、HX008(翰中/翰思/康方公司)、AK105(康方公司)、LZM009(丽珠公司)、HLX10(复宏汉霖公司)。
附图说明
图1显示不同剂量的化合物6和E7046在CT-26结肠直肠癌模型中对肿瘤生长的影响。
图2显示不同剂量的化合物6和E7046在CT-26结肠直肠癌模型中对小鼠体重的影响。
图3显示不同剂量的化合物6和抗PD-1抗体在CT-26结肠直肠癌模型中分别单独、联合施用时对肿瘤生长的影响。
图4显示不同剂量的化合物6和抗PD-1抗体在CT-26结肠直肠癌模型中单独和联合施用时对小鼠体重的影响。
图5显示不同剂量的化合物6和抗PD-1抗体在EMT-6乳腺癌模型中单独和联合施用时对肿瘤生长的影响。
图6显示不同剂量的化合物6和抗PD-1抗体在乳腺癌肿瘤模型中单独和联合施用时对小鼠体重的影响。
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)，氘代二甲亚砜(DMSO-d 6)，氘代甲醇(CD 3OD)或者氘代乙腈(CD 3CN)。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，q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：SHIMADZU LCMS-2020，色谱柱：
Figure PCTCN2020072487-appb-000019
5μm x 30x2.1mm S/N：H17-247175，柱温：50.0℃，流动相：A：水(0.0375％TFA)，B：乙腈(0.01875％TFA)，电离模式：ESI，极性：正。
梯度洗脱
时间(分钟) B(％) 流速(mL/分钟)
0.0 5 1.5
0.80 95 1.5
1.20 95 1.5
1.21 5 1.5
1.55 5 1.5
核磁共振仪：Bruker ARX-500型高分辨质谱和Bruker ARX-400型高分辨质谱。
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)。
常用缩写
EP4:     前列腺素E2受体4
PGE2:    前列腺素E2
COX:     环加氧酶
cAMP:    环磷酸腺苷
PKA:     蛋白激酶A
PI3K:    还刺激非经典途径磷脂酰肌醇3激酶
ERK:     细胞外调节激酶
NSAIDs:  非甾体类抗炎药
TNF-α:  肿瘤坏死因子α
IL-12:   白细胞介素12
NMR:     核磁共振
MS:      质谱
DMSO:    二甲亚砜
DMF：    二甲基甲酰胺
TLC:     薄层色谱法
HATU:    1-[双(二甲基氨基)亚甲基]-1H-1,2,3-三唑并[4,5-b]吡啶-3-氧化六氟磷酸盐
DIPEA:   二异丙基乙胺
LCMS:    液相色谱-质谱
HPLC:    高效液相色谱
Rt:      保留时间
DME:     二甲氧基乙烷
NBS:     N-溴琥珀酰亚胺
AIBN:    偶氮二异丁腈
DCM:     二氯甲烷
Dppf:    二苯基膦铁氧体
TEA:     三乙胺
CDI:     羰基
TFAA:    三氟乙酸酐
BPO:     过氧化苯甲酰
THF:     四氢呋喃
Xphos:   2-二环己基膦基-2',4',6'-三异丙基
tBuBrettPhos：2-(二叔丁基膦基)-2',4',6'-三异丙基-3,6-二甲氧基-1,1'-联苯
FDD(First Dose Double)：第一个剂量加倍
实施例1
3-(1-(1-(4-(三氟甲基)苄基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸1
Figure PCTCN2020072487-appb-000020
步骤1：1-(4-(三氟甲基)苄基)-1H-吲哚-7-羧酸甲酯1-2的制备
将1H-吲哚-7-甲酸甲酯1-1(5.00g，28.5mmol)和1-(溴甲基)-4-(三氟甲基)苯(7.75g，32.4mmol，5mL)溶于DMF(130mL)中，在冰浴冷却下缓慢加入NaH(1.26g，31.3mmol，60％纯度)。将得到的混合物继续搅拌3小时，然后除去冰浴，将得到的混合物在20℃下搅拌6小时。TLC(石油醚/乙酸乙酯＝5/1)显示原料(Rf＝0.40)被消耗完并形成主要的新点(Rf＝0.5)后，将反应混合物用饱和NH 4Cl(100mL)水溶液淬灭，然后用乙酸乙酯(100mL)萃取。将有机层用盐水(200mL)洗涤，用无水硫酸钠干燥，过滤并浓缩，将得到的残余物通过快速硅胶色谱法纯化(
Figure PCTCN2020072487-appb-000021
20g
Figure PCTCN2020072487-appb-000022
硅胶柱，0～3％的乙酸乙酯/石油醚梯度洗 脱液),得到化合物1-2(6.50g，18.7mmol，65.5％收率)，为黄色固体。MS(ESI):334.1[M+1] +。
步骤2：1-(4-(三氟甲基)苄基)-1H-吲哚-7-羧酸1-3的制备
向化合物1-2(3.00g，9.00mmol)的甲醇(45mL)和四氢呋喃(45mL)溶液中加入KOH水溶液(2M，45mL)，并将所得混合物在25℃下搅拌12小时。LCMS显示起始材料消耗完并且检测到具有所需产品分子量的峰(Rt＝0.948min)后，将反应混合物在45℃下浓缩以除去大部分甲醇和四氢呋喃，用1N盐酸水溶液酸化至pH～6-7，然后用1N盐酸洗涤并用乙酸乙酯(100mL)萃取。将有机层用无水硫酸钠干燥，过滤并浓缩，得到化合物1-3(2.20g，6.89mmol，76.5％收率)，为黄色固体。MS(ESI):320.1[M+1] +。
制备3-(1-氨基环丙基)双环[1.1.1]戊烷-1-羧酸甲酯A-1的一般方法
Figure PCTCN2020072487-appb-000023
在氮气保护及-20℃的温度下，向3-氰基双环[1.1.1]戊烷-1-羧酸甲酯(22.5g，99.2mmol)的甲苯(240mL)溶液中加入四异丙氧基钛(29.8g，99.7mmol，31mL，95％纯度)。然后在氮气保护及-20℃的温度下，在30分钟内逐滴加入EtMgBr(3M，60mL)，保持温度在-20～-10℃之间。搅拌30分钟后，逐滴加入BF 3·Et 2O(27.6g，194mmol，24mL)并在-20℃下搅拌30分钟，然后在25℃下搅拌12小时。TLC(板1：石油醚/乙酸乙酯＝3/1)显示原料(Rf＝0.61)已经消耗完毕，TLC(板2：石油醚/乙酸乙酯＝1/1)显示有一个主要产品(Rf＝0.24)形成。在0℃下缓慢加入盐酸水溶液(1N,30mL)将反应混合物淬灭，然后弃去分离的有机层。在0℃下用10M氢氧化钠水溶液将水相碱化至pH～12并用乙酸乙酯(200mL×2)萃取。将合并的有机层浓缩，得到残余物，将其通过柱色谱(二氧化硅，石油醚/乙酸乙酯＝20/1至1/1)纯化，得到化合物A-1(3.9g，21.7mmol，21.9％收率)，为黄色固体。MS(ESI):182.3[M+1] +。
步骤3：3-(1-(1-(4-(三氟甲基)苄基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊 烷-1-羧酸甲酯1-4的制备
向化合物1-3(0.65g，2.04mmol)，化合物A-1(378mg，2.09mmol)和HATU(814mg，2.14mmol)的DMF(5mL)溶液中加入DIPEA(742mg，5.74mmol，1.0mL)。将所得混合物在氮气下在20℃下搅拌8小时。LCMS显示起始化合物1-3被消耗，并且检测到具有所需质量的一个主产品峰(Rt＝0.947min)。将反应混合物用乙酸乙酯(40mL)稀释，并用盐水(50mL×2)洗涤。将有机层用无水硫酸钠干燥，过滤并减压浓缩，得到残余物，将其通过柱色谱(二氧化硅，石油醚/乙 酸乙酯＝100/1到3/1，Rf＝0.40)纯化，获得化合物1-4(0.70g，粗品)，为白色固体。MS(ESI):483.1[M+1] +。
步骤4：3-(1-(1-(4-(三氟甲基)苄基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊 烷-1-羧酸1的制备
向化合物1-4(0.70g，1.50mmol)在乙醇(10mL)的溶液中加入LiOH·H 2O(4M，1mL)，将得到的混合物在65℃下搅拌24小时。LCMS显示原料消耗完并且检测到具有所需产品分子量的主峰。将反应混合物在50℃下浓缩以除去大部分乙醇。加入30mL水，用1N盐酸酸化至pH～5，用乙酸乙酯(3×50mL)萃取。合并有机层，用无水硫酸钠干燥，过滤并浓缩，得到残余物，将其通过HPLC制备(柱：Phenomenex Synergi C18 150×25×10μm；流动相：[水(0.225％甲酸)-乙腈]；B％：50％-77％，10分钟)得到化合物1(350mg，747μmol，52％收率)，白色固体。MS(ESI):469.2[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ12.21(br s,1H),8.55(s,1H),7.72(dd,J＝1.1,7.8Hz,1H),7.56(d,J＝8.2Hz,2H),7.47(d,J＝3.2Hz,1H),7.19(dd,J＝1.0,7.2Hz,1H),7.12-7.07(m,1H),6.96(d,J＝8.2Hz,2H),6.64(d,J＝3.2Hz,1H),5.70(s,2H),1.70(s,6H),0.57-0.51(m,2H),0.34-0.27(m,2H)。
实施例2
3-(1-(1-((5-苯基吡啶-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸2
Figure PCTCN2020072487-appb-000024
步骤1：2-甲基-5-苯基吡啶2-2的制备
在氮气下，向5-溴-2-甲基吡啶2-1(15.0g，87.2mmol)的DME(150mL)溶液中加入Pd(PPh 3) 4(5.04g，4.36mmol)，4,4,5,5-四甲基-2-苯基-1,3,2-二氧硼戊环(20.0g，98.0mmol)和K 2CO 3(2M，66.0mL)，将所得混合物在120℃下搅拌6小时。TLC(石油醚/乙酸乙酯＝5/1)表明原料(Rf＝0.6)被消耗并形成新的点(Rf＝0.37)。用乙酸乙酯(100mL)萃取反应混合物。将有机层用盐水(200mL)洗涤，经无水硫酸钠干燥，过滤并在减压下浓缩，得到残余物，将其通过快速硅胶色谱法纯化(
Figure PCTCN2020072487-appb-000025
10g
Figure PCTCN2020072487-appb-000026
二氧化硅柱，洗脱液为2～10％乙酸乙酯/石油醚梯度，Rf＝0.46)，得到化合物2-2(10.0g，59.0mmol，67.7％收率)，为黄色液体。MS(ESI):170[M+1] +。
1H NMR(400MHz,CDCl 3):δ8.75(d,J＝2.3Hz,1H),7.79(dd,J＝2.4,8.0Hz,1H),7.61-7.55(m,2H),7.51-7.44(m,2H),7.43-7.35(m,1H),7.23(d,J＝7.9Hz,1H),2.62(s,3H)。
步骤2：2-(溴甲基)-5-苯基吡啶2-3的制备
向化合物2-2(5.00g，29.5mmol)和NBS(5.26g，29.5mmol)的CCl 4(100mL)溶液中加入苯甲酰基苯甲酸过氧化物(720mg，2.97mmol)。将所得混合物在75℃下搅拌10小时。TLC(石油醚/乙酸乙酯＝5/1)表明大部分起始材料(Rf＝0.45)被消耗并形成主要的新点(Rf＝0.57)。将反应混合物在50℃下浓缩，得到残余物，将其通过快速硅胶色谱法(
Figure PCTCN2020072487-appb-000027
10g
Figure PCTCN2020072487-appb-000028
二氧化硅柱，0～3％乙酸乙酯/石油醚梯度)纯化，得到化合物2-3(1.50g，5.85mmol，19.8％收率)，为粉红色固体。MS(ESI):248[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ8.93-8.83(m,1H),8.11(dd,J＝2.4,8.1Hz,1H),7.78-7.72(m,2H),7.65(dd,J＝0.6,8.1Hz,1H),7.55-7.49(m,2H),7.48-7.42(m,1H),4.76(s,2H)。
步骤3：1-((5-苯基吡啶-2-基)甲基)-1H-吲哚-7-羧酸甲酯2-4的制备
按照与制备化合物1-2类似的方法，使1H-吲哚-7-羧酸甲酯1-1与化合物2-3反应。得到所需化合物2-4(1.50g，4.38mmol，72.4％收率)，为黄色固体。MS(ESI):343.2[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ8.76(d,J＝1.8Hz,1H),7.94(dd,J＝2.3,8.2Hz,1H),7.83(d,J＝7.8Hz,1H),7.69-7.61(m,3H),7.51-7.35(m,4H),7.10(t,J＝7.6Hz,1H),6.75-6.66(m,2H),5.74(s,2H),3.70(s,3H)。
步骤4：1-((5-苯基吡啶-2-基)甲基)-1H-吲哚-7-羧酸2-5的制备
按照制备化合物1-3相似的方法，水解化合物2-4，得到化合物2-5(0.80g，粗品)，为黄色固体，将其直接用于下一步骤。MS(ESI):329.2[M+1] +。
步骤5：3-(1-(1-((5-苯基吡啶-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环 [1.1.1]戊烷-1-羧酸甲酯2-6的制备
该反应按照与制备化合物1-4类似方法进行，得到化合物2-6(0.50g，粗品)，为白色固体。MS(ESI):429.3[M+1] +。
步骤6：3-(1-(1-((5-苯基吡啶-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环 [1.1.1]戊烷-1-羧酸2的制备
该反应按照与制备化合物1类似方法进行，得到化合物2，为白色固体。MS(ESI):478.1[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ8.84(br s,1H),8.52(s,1H),8.04-7.92(m,1H),7.73(d,J＝7.9Hz,1H),7.69-7.63(m,2H),7.55-7.47(m,3H),7.46-7.39(m,2H),7.22-7.16(m,1H),7.14-7.07(m,1H),6.67(d,J＝3.1Hz,1H),6.42(br d,J＝8.1Hz,1H),5.76(br s,2H),1.72(s,6H),0.659-0.56(m,2H),0.41-0.38(m,2H)。
实施例3
3-(1-(1-(喹啉-2-基甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸3
Figure PCTCN2020072487-appb-000029
按照与实施例1类似的制备方法，由1-(喹啉-2-基甲基)-1H-吲哚-7-羧酸甲酯替换化合物1-2，得到化合物3，为淡黄色固体。MS(ESI):452.1[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ8.49(s,1H),8.26(br,1H),8.04-7.90(m,2H),7.83-7.70(m,2H),7.66-7.53(m,2H),7.20-7.15(m,1H),7.13-7.06(m,1H),6.68(d,J＝3.2Hz,1H),6.52(d,J＝8.6Hz,1H),5.87(s,2H),4.2(br,1H),1.69(s,6H),0.47-0.44(m,2H),0.29-0.22(m,2H)。
实施例4
3-(1-(1-(4-氯苄基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸4
Figure PCTCN2020072487-appb-000030
按照实施例1中描述的方法，由1-(4-氯苄基)-1H-吲哚-7-羧酸甲酯替换化合物1-2，得到化合物4，为灰白色固体。MS(ESI):434.1[M+1] +。
1H NMR(400MHz,CDCl 3):δ7.77(dd,J＝0.9,7.9Hz,1H),7.23-7.14(m,3H),7.14-7.07(m,2H),6.72(d,J＝8.4Hz,2H),6.63(d,J＝3.2Hz,1H),6.06(s,1H),5.60(s,2H),1.94(s,6H),0.76-0.68(m,2H),0.52-0.46(m,2H)。
实施例5
3-(1-(1-((6-苯基吡啶-3-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸5
Figure PCTCN2020072487-appb-000031
步骤1：5-甲基-2-苯基吡啶5-2的制备
在氮气气氛下,向2-溴-5-甲基吡啶5-1(4.00g，23.2mmol)和4,4,5,5-四甲基-2-苯基-1,3,2-二氧杂硼杂环戊烷a(5.31g，26.0mmol)的DME(40.0mL)溶液中加入K 2CO 3水溶液(2.00M，17.5mL)和Pd(PPh 3) 4(1.34g，1.16mmol)。将所得混合物在110℃下搅拌15小时。TLC(石油醚/乙酸乙酯＝20/1)显示原料(Rf＝0.55)完全消耗并形成主要新点(Rf＝0.50)。将反应混合物冷却至室温并用乙酸乙酯(20.0mL×3)萃取。将合并的有机层用水(20.0mL×2)和盐水(20.0mL)洗涤，用无水硫酸钠干燥，过滤并在真空下浓缩，得到残余物，将其通过柱色谱纯化(二氧化硅，石油醚/乙酸乙酯＝1/0～30/1)得到化合物5-2(4.50g，19.8mmol，57.0％收率)为黄色油状物。MS(ESI):170.1[M+1] +。
步骤2：5-甲基-2-苯基吡啶5-3的制备
在氮气气氛下，向含有化合物5-2(3.00g，13.2mmol)和NBS(2.83g，15.9mmol)的CCl 4(30.0mL)溶液中加入AIBN(217mg，1.33mmol)。将所得混合物在氮气氛下在70℃下搅拌14小时。TLC(石油醚/乙酸乙酯＝20/1)显示大部分原料(Rf＝0.40)被消耗并且形成主要新点(Rf＝0.33)。向反应混合物中加入水(20.0mL)并用CH 2Cl 2(20mL×3)萃取。合并的有机层用盐水(20.0mL×2)洗涤，用无水硫酸钠干燥，过滤并真空浓缩，得到残余物，通过柱色谱(二氧化 硅，石油醚/乙酸乙酯＝1/0～20/1)得到化合物5-3(1.80g，4.81mmol，36.2％收率)，为浅黄色固体。MS(ESI):248.1[M+1] +。
步骤3-6：3-(1-(1-((6-苯基吡啶-3-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环 [1.1.1]戊烷-1-羧酸5的制备
从化合物5-3到最终产物的转化与实施例2中使用的方法相似，得到的化合物5为灰白色固体。MS(ESI):478.2[M+1] +。
1H NMR(400MHz,CDCl 3):δ12.24(s,1H),8.73(d,J＝1.6Hz,1H),8.15(d,J＝1.6Hz,1H),7.89(dd,J＝8.0Hz,2H),7.78(dd,J＝8.0Hz,1H),7.59(d,J＝8.0Hz,1H),7.44-7.39(m,3H),7.26-7.22(m,1H),7.17-7.16(d,J＝3.2Hz,1H),7.12-7.09(t,J＝7.6Hz,1H),6.66-6.65(m,1H),6.21(s,1H),5.69(s,2H),1.92(s,6H),0.72-0.70(m,2H),0.53-0.52(m,2H)。
实施例6
3-(1-(1-(苯并呋喃-2-基甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸6
Figure PCTCN2020072487-appb-000032
步骤1：苯并呋喃-2-基甲醇6-1的合成
在反应瓶中，依次加入苯并呋喃-2-甲醛(3g，20.5mmol)和无水甲醇(40ml)，冷却至0℃，将硼氢化钠(0.545g，14.4mmol)分批加入其中,保持温度低于25℃。加料完毕，在室温下搅拌1小时。反应完毕，减压蒸除溶剂，加入1N HCl水溶液(15ml)，室温搅拌5分钟，然后用饱和碳酸氢钠水溶液调pH至8-9，乙酸乙酯(10ml x 3)萃取，合并的有机相用饱和食盐水水洗，无水硫酸钠干燥，减压蒸除溶剂，得化合物6-1(3.0g，20.27mmol，收率：98.9％)为黄色油状物。MS(ESI):149.1[M+1] +。
1H NMR(400MHz,CDCl 3)δ:7.55(dd,J＝8.4,7.2Hz,1H),7.47(dd,J＝8.8,7.6Hz,1H),7.29-7.19(m,2H),6.66(s,1H),4.77(d,J＝4.8Hz,2H)。
步骤2：2-(溴甲基)苯并呋喃6-2的合成
在反应瓶中，依次加入化合物6-1(2.47g，16.7mmol)和干燥的二氯甲烷(32ml)，冷却至0℃。将三溴化磷(1.72mL，18.4mmol)缓慢滴加其中，加料完毕，升温至室温搅拌1小时。经TLC监测(石油醚：乙酸乙酯＝9：1)反应完毕，用饱和碳酸氢钠水溶液调pH至8-9，二氯甲烷(10ml x 3)萃取，合并的有机相用饱和食盐水水洗，无水硫酸钠干燥，减压蒸除溶剂，得化合物6-2(3.36g，收率：95.9％)为黄色油状物。
然后，按照实施例1中描述的方法，由化合物6-2替换1-(溴甲基)-4-(三氟甲基)苯，得到3-(1-(1-(苯并呋喃-2-基甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸6，为灰白色固体。MS(ESI):441.1[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ12.21(s,1H),8.73(s,1H),7.69(d,J＝7.6Hz,1H),7.51(d,J＝3.2Hz,1H),7.45(t,J＝7.8Hz,2H),7.24-7.06(m,4H),6.60(d,J＝3.2Hz,1H),6.18(s,1H),5.76(s,2H),1.76(s,6H),0.63(d,J＝7.2Hz,2H),0.50(t,J＝5.6Hz,2H)。
实施例7
3-(1-(5-氯-1-((5-苯基吡啶-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸7
Figure PCTCN2020072487-appb-000033
步骤1：5-氯-1H-吲哚-7-羧酸甲酯7-2的制备
向7-溴-5-氯-1H-吲哚7-1(1.40g，6.07mmol)的甲醇(50.0mL)溶液中加入Pd(dppf)Cl 2CH 2Cl 2(98.0mg，120μmol)，dppf(112mg，202μmol)和TEA(2.80 mL)。将所得混合物用一氧化碳吹扫并在CO气氛(4.0MPa)下在130℃下搅拌48小时。LC-MS显示原料完全消耗，检测到所需产平的分子质量(RT＝0.880)。减压浓缩混合物，得到残余物，通过柱色谱(二氧化硅，石油醚/乙酸乙酯＝1/0～20/1梯度洗脱，Rf＝0.55)纯化，得到化合物7-2(900mg，4.29mmol,70.7％收率)，为黄色固体。MS(ESI):210.0[M+1] +。
1H NMR(400MHz,CDCl 3):δ9.85(s,1H),7.87(d,1H,J＝2.0Hz),7.84(d,1H,J＝1.8Hz),7.4-7.4(m,1H),6.6-6.6(m,1H),4.02(s,3H)。
步骤2-5：3-(1-(5-氯-1-((5-苯基吡啶-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基) 双环[1.1.1]戊烷-1-羧酸7的制备
按照实施例1中描述的方法，将中间体7-2转化为灰白色固体的最终产物。MS(ESI):512.1[M+1] +。
1H NMR(400MHz,DMSO-d 6):δ12.24(s,1H),8.73(d,J＝1.6Hz,1H),8.63(s,1H),7.85(dd,J＝2.2,8.2Hz,1H),7.78(d,J＝2.1Hz,1H),7.66-7.57(m,3H),7.47(t,J＝7.5Hz,2H),7.43-7.35(m,1H),7.10(d,J＝1.8Hz,1H),6.65(d,J＝3.2Hz,1H),6.41(d,J＝8.2Hz,1H),5.69(s,2H),1.66(s,6H),0.55-0.48(m,2H),0.36-0.33(m,2H)。
实施例8
3-(1-(5-氯-1-(4-(三氟甲基)苄基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸8
Figure PCTCN2020072487-appb-000034
按照实施例1中描述的方法，由5-氯-1-(4-(三氟甲基)苄基)-1H-吲哚-7-羧酸甲酯替换化合物1-2，得到为白色固体的化合物8。MS(ESI):503.1[M+1] +。
1H NMR(400MHz,DMSO-d 6):δ8.45(s,1H),8.12(s,1H),7.55(d,1H,J＝2.1Hz),7.34(s,1H),7.31(s,1H),7.30(d,1H,J＝3.2Hz),6.89(d,1H,J＝2.1Hz),6.71(s,1H),6.69(s,1H),6.40(d,1H,J＝3.2Hz),5.43(s,2H),1.43(s,6H),0.3-0.3(m,2H),0.0-0.0(m,2H)。
实施例9
4-(1-(1-(4-(三氟甲基)苄基)-1H-吲哚-7-甲酰胺基)环丙基)双环[2.2.2]辛烷-1-羧酸9
Figure PCTCN2020072487-appb-000035
步骤1：4-氨基甲酰基双环[2.2.2]辛烷-1-羧酸甲酯9-2的制备
向4-(甲氧基羰基)双环[2.2.2]辛烷-1-羧酸9-1(5.00g，23.5mmol)的二氯甲烷(50mL)溶液中加入CDI(4.21g，25.9mmol)，所得混合物在25℃下搅拌1小时。然后加入DIEA(3.99g，30.8mmol)和NH 4Cl(1.51g，28.2mmol)。将所得混合物在25℃下搅拌2小时。TLC(石油醚/乙酸乙酯＝1/1，I 2)显示原料(Rf＝0.50)被消耗并形成一个主要的新点(Rf＝0.45)。反应混合物用1N盐酸水溶液将其酸化至pH～3。用二氯甲烷(30mL×3)萃取。将合并的有机相用盐水(50mL)洗涤，用无水硫酸钠干燥，过滤并在真空下浓缩，得到粗制的化合物9-2(4.00g，18.9mmol)，80.3％收率)，为白色固体。MS(ESI):211.1[M+1] +。不经纯化直接用于下一步。
步骤2：4-氰基双环[2.2.2]辛烷-1-羧酸甲酯9-3的制备
在0°下，向化合物9-2(4.00g，18.9mmol)的吡啶(49.0g，619mmol)溶液中加入TFAA(9.06g，43.1mmol)。将所得混合物在25℃下搅拌1小时。TLC(石油醚/乙酸乙酯＝3/1)显示原料(Rf＝0.25，板1)被消耗，并且检测到一个新的点(Rf＝0.56，板2)。用5.0N盐酸水溶液将其酸化至pH＝2～3。用二氯甲烷(50mL×3)萃取。将合并的有机相用盐水(80mL×3)洗涤，用无水硫酸钠干燥，过滤并在真空下浓缩得到残余物，将其通过柱色谱法纯化(石油醚/乙酸乙酯＝50/1～3/1)得到化合物9-3(1.10g，5.69mmol，30.0％收率)，为白色固体。MS(ESI):194.1[M+1] +。
1H NMR(400MHz,CDCl 3):δ3.66(s,3H),2.03-1.92(m,6H),1.89-1.77(m,6H)。
步骤3：4-(1-氨基环丙基)双环[2.2.2]辛烷-1-羧酸甲酯9-4的制备
在25℃下向化合物9-3(800mg，4.14mmol)的甲苯(15mL)溶液中滴加Ti(O-Et) 4(1.05g，4.58mmol)。将所得混合物在25℃下搅拌15分钟，然后在-20℃下滴加EtMgBr(3M，2.76mL)并将混合物在-20℃下搅拌15分钟。在-20℃下滴加BF 3·Et 2O(1.20g，8.43mmol)，在25℃下搅拌24小时.LC-MS显示原料消耗并且 检测到所需分子量(检测到RT＝0.838)。向混合物中加入10毫升1N盐酸水溶液和15mL水，用乙酸乙酯(15mL×3)洗涤。用10M NaOH水溶液将水溶液碱化至pH＝8～9。用乙酸乙酯(25mL×3)萃取。将合并的有机相用盐水(50mL)洗涤，用无水硫酸钠干燥，过滤并真空浓缩，得到残余物。将残余物直接用于下一步，无需纯化。得到化合物9-4(0.200g，粗品)，为黄色油状物。MS(ESI):224.1[M+1] +。
步骤4：4-(1-(1-(4-(三氟甲基)苄基)-1H-吲哚-7-甲酰胺基)环丙基)双环[2.2.2]辛 烷-1-羧酸甲酯9-5的制备
向化合物1-3(120mg，375μmol)的DMF(2.00mL)溶液中加入化合物9-4(92.3mg，413μmol)，HATU(160mg，420μmol)和DIPEA(185μL，1.06mmol)。将所得混合物在25℃下搅拌1小时。LC-MS显示原料化合物1-3被消耗并且检测到所需产品分子量(RT＝1.051)。加入10mL水并用乙酸乙酯(1 0mL×3)萃取。合并的有机相用盐水(20mL×2)洗涤，用无水硫酸钠干燥，过滤并真空浓缩得到残余物，用柱色谱法纯化(石油醚/乙酸乙酯＝50/1～3/1，板1：石油醚/乙酸乙酯＝3/1，Rf＝0.45)得到化合物9-5(140mg，236μmol，62.9％收率)为白色固体。MS(ESI):525.2[M+1] +。
步骤5：4-(1-(1-(4-(三氟甲基)苄基)-1H-吲哚-7-甲酰胺基)环丙基)双环[2.2.2]辛 烷-1-羧酸9的制备
向化合物9-5(140mg，236μmol)的甲醇(2.00mL)溶液中加入LiOH·H 2O水溶液(4M，1.86mL)并将混合物在60℃下搅拌1小时。LC-MS显示原料耗尽并且检测到所需产品分子量(RT＝1.022)。浓缩反应混合物以除去甲醇。加入9mL1N盐酸水溶液。用乙酸乙酯(15mL×3)萃取。将合并的有机相真空浓缩，得到残余物，将其通过制备HPLC(柱：Shim-pack C18 150×25×10μm；流动相：[水(0.225％甲酸)-乙晴]；B％：45％-75％，10min)纯化得到化合物9(75.6mg，146μmol，61.9％收率)，为浅绿色固体。MS(ESI):511.2[M+1] +。
1H NMR(400MHz,CDCl 3):δ7.78(dd,J＝1.2,7.9Hz,1H),7.48(d,J＝8.2Hz,2H),7.22(dd,J＝1.0,7.2Hz,1H),7.16-7.10(m,2H),6.86(d,J＝7.9Hz,2H),6.66(d,J＝3.2Hz,1H),5.95(s,1H),5.74(s,2H),1.77-1.72(m,6H),1.46-1.41(m,6H),0.78-0.72(m,2H),0.34-0.28(m,2H)。
实施例10
3-(1-(1-((5-吗啉代吡啶-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸10
Figure PCTCN2020072487-appb-000036
步骤1：5-溴-2-(溴甲基)吡啶10-2的制备
向5-溴-2-甲基吡啶10-1(20.0g，116mmol)的四氯甲烷(200mL)溶液中加入NBS(22.7g，127mmol)和BPO(1.30g，5.35mmol)。将所得混合物在76℃下搅拌12小时。TLC(石油醚/乙酸乙酯＝30/1)表明化合物10-1(Rf＝0.6)被消耗，形成两个新斑点(Rf＝0.65，Rf＝0.7)。将反应混合物用水(70.0ml×3)洗涤，用无水硫酸钠干燥，过滤并浓缩，得到残余物，通过柱色谱(二氧化硅，石油醚/乙酸乙酯＝1/0-5/1，Rf＝0.65)纯化。通过LCMS确认化合物10-2(9.00g，30.3％产率，98.3％纯度)，为浅黄色油状物。MS(ESI):249.1[M+1] +。
1H NMR(400MHz,DMSO-d 6):δ8.55(d,J＝2.32Hz,1H),7.74(dd,J＝8.31,2.45Hz,1H),7.28(d,J＝8.31Hz,1H),4.45(s,2H)。
步骤2：1-((5-溴吡啶-2-基)甲基)-1H-吲哚-7-羧酸甲酯10-3的制备
在0℃和氮气下，向甲基1H-吲哚-7-羧酸甲酯1-2A(7.54g，43.0mmol)的DMF(40.0mL)溶液中加入NaH(1.03g，43.0mmol)。将所得混合物在25℃下搅拌30分钟。然后在0℃和氮气下加入5-溴-2-(溴甲基)吡啶10-2(9.00g，35.8mmol)的DMF(40.0mL)溶液。将所得混合物在25℃下搅拌30分钟。LC-MS显示起始化合物1-2A完全消耗。将反应混合物用水(100mL)稀释，用乙酸乙酯(300mL×2)萃取。将合并的有机层用盐水(100mL×2)洗涤，用无水硫酸钠干燥，过滤并浓缩，得到残余物，将其通过柱色谱(二氧化硅，石油醚/乙酸乙酯＝1/0至2/1)纯化。得到化合物10-3(6.80g，54.4％收率，99.0％纯度)，为棕色固体。MS(ESI):345.1[M+1] +。
1H NMR(400MHz,CDCl3)：δ8.52(d,J＝2.1Hz,1H),7.74(dd,J＝1.2,7.9Hz, 1H),7.60-7.50(m,2H),7.13(d,J＝3.2Hz,1H),7.06(t,J＝7.6Hz,1H),6.60(d,J＝3.3Hz,1H),6.44(d,J＝8.3Hz,1H),5.61(s,2H),3.68(s,3H)。
步骤3：1-((5-溴吡啶-2-基)甲基)-1H-吲哚-7-羧酸10-4的制备
向化合物10-3(6.8g，19.7mmol)的甲醇(90.0mL)和THF(90.0mL)溶液中加入LiOH·H 2O(9.9g，176mmol)的水(90.0mL)溶液。将所得混合物在25℃下搅拌12小时。LC-MS显示化合物10-3完全消耗，检测到所需质量(RT＝0.861)。浓缩反应混合物以除去甲醇和THF。用盐酸(1M)水溶液将其酸化至pH＝5～6，大量固体沉淀。将其过滤，收集固体，得到化合物10-4(4.00g，61.3％产率)，为白色固体。MS(ESI):331.1[M+1] +。
1H NMR(400MHz,DMSO-d 6):δ8.57(s,1H),7.87-7.93(m,1H),7.78-7.83(m,1H),7.57(d,J＝3.30Hz,1H),7.47-7.53(m,1H),7.05-7.14(m,1H),6.65-6.70(m,1H),6.48-6.56(m,1H),5.75-5.84(m,2H)。
步骤4：1-((5-吗啉代吡啶-2-基)甲基)-1H-吲哚-7-羧酸10-5的制备
向化合物10-4(240mg，724μmol)的二氧六环(12.0mL)溶液中加入吗啉(126mg，1.45mmol，127μL)，XPhos(69.1mg，144μmol)和Cs 2CO 3(944mg，2.90mmol)。将所得混合物脱气并用氮气吹扫3次，加入Pd 2(dba) 3(66.3mg，72.4μmol)并将混合物在氮气下在120℃下搅拌2小时。LCMS显示化合物10-4完全消耗，检测到所需质量(RT＝0.233)。过滤混合物，浓缩滤液，得到残余物，将其通过制备HPLC(中性)纯化，得到化合物10-5(12 0mg，41.9％收率，85.4％纯度)，为浅黄色固体。MS(ESI):337.1[M+1] +。
步骤5：3-(1-(1-((5-吗啉代吡啶-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环 [1.1.1]戊烷-1-羧酸甲酯10-6的制备
将化合物10-5(50.0mg，148μmol)，3-(1-氨基环丙基)双环[1.1.1]戊烷-1-羧酸甲酯A1(27.0mg，149μmol)，HATU(57.0mg，149μmol)和DIEA(57.8mg，447μmol，78.0μL)在DMF(1.0mL)中的混合物在25℃下搅拌4小时。LCMS显示化合物10-5完全消耗，检测到所需质量(RT＝0.792)。TLC(乙酸乙酯)表明化合物10-5(Rf＝0.30)被完全消耗，形成两个新斑点(Rf＝0.41，Rf＝0.6)。将反应混合物用水(5mL)稀释，用乙酸乙酯(10ml×2)萃取。将合并的有机层用盐水(20ml×2)洗涤，用无水硫酸钠干燥，过滤并浓缩，得到残余物，将其通过制备TLC(二氧化硅，乙酸乙酯，Rf＝0.6)纯化，得到化合物10-6(60.0mg，80.8％产率)，为白色固体。MS(ESI):501.1[M+1] +。
步骤6：3-(1-(1-((5-吗啉代吡啶-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环 [1.1.1]戊烷-1-羧酸甲酯10的制备
向化合物10-6(60.0mg，119μmol)的乙醇(3.00mL)溶液中加入LiOH·H 2O(49.0mg，1.17mmol)的水(0.3mL)溶液。将所得混合物在65℃下搅拌12小时。LCMS显示化合物10-6完全消耗，检测到所需质量(RT＝0.777)。将反应混 合物酸化至pH＝6to 7。通过制备HPLC(柱：Phenomenex Synergi C18 150×25×10μm；流动相：[水(0.225％FA)-ACN]；B％：12％-42％，10分钟)直接纯化，得到化合物10(16.6mg，28.3％产率)纯度为99.4％，为浅黄色固体。MS(ESI):487.2[M+1] +。
1H NMR(400MHz,DMSO-d 6):δ12.24(s,1H),8.50(s,1H),8.17(d,J＝2.81Hz,1H),7.67(dd,J＝7.83,1.22Hz,1H),7.41(d,J＝3.18Hz,1H),7.10-7.15(m,2H),7.08-7.03(m,1H),6.59(d,J＝3.18Hz,1H),6.30(d,J＝8.68Hz,1H),5.52(s,2H),3.73-3.69(m,4H),3.08-3.05(m,4H),1.70(s,6H),0.62-0.58(m,2H),0.50-0.45(m,2H)。
实施例11
3-(1-(1-([2,3'-联吡啶]-6'-基甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸11
Figure PCTCN2020072487-appb-000037
步骤1：1-([2,3'-联吡啶]-6'-基甲基)-1H-吲哚-7-羧酸11-1的制备
向化合物10-4(200mg，603μmol)，吡啶-2-基硼酸(80.0mg，650μmol)和Na 2CO 3(212.0mg，2.00mmol)在水(1.5mL)和二氧六环(6mL)溶液中加入Pd(PPh 3) 4(44.0mg，38.0μmol)。将所得混合物在氮气氛下在100℃下搅拌8小时。LCMS显示化合物10-4被消耗并且检测到所需质量(RT＝0.794)。过滤反应混合物，减压浓缩滤液，得到残余物，通过反相HPLC(0.1％FA条件)纯化，得到化合物11-1(120mg，收率29.6％，纯度96.2％)为白色固体。MS(ESI):330.3[M+1] +。
步骤2：3-(1-(1-([2,3'-联吡啶]-6'-基甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环 [1.1.1]戊烷-1-羧酸甲酯11-2的制备
将化合物11-1(50.0mg，150μmol)，3-(1-氨基环丙基)双环[1.1.1]戊烷-1-羧酸甲酯A1(27.0mg，149μmol)，HATU(57.0mg，149μmol)和DIEA(57.8mg，447μmol，78.0μL)在DMF(1.0mL)中的混合物)在25℃下搅拌4小时。LCMS显示化合物11-1完全消耗，检测到所需质量(RT＝0.692)。将反应混合物用水(5 mL)稀释，用乙酸乙酯(10ml×2)萃取。将合并的有机层用盐水(20ml×2)洗涤，用无水硫酸钠干燥，过滤并浓缩，得到残余物，将其通过制备TLC(SiO2，乙酸乙酯，Rf＝0.5)纯化，得到化合物11-2(58.0mg，79％产率)，为白色固体。MS(ESI):493.1[M+1] +。
步骤3：3-(1-(1-([2,3'-联吡啶]-6'-基甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环 [1.1.1]戊烷-1-羧酸11的制备
向化合物11-2(58.0mg，117μmol)的乙醇(3.00mL)溶液中加入LiOH·H 2O(49.0mg，1.17mmol)的水(0.3mL)溶液。将所得混合物在65℃下搅拌12小时。LCMS显示化合物11-2完全消耗，检测到所需质量(RT＝0.777)。将反应混合物酸化至pH＝6to 7。通过制备HPLC(柱：Phenomenex Synergi C18 150×25×10μm；流动相：[水(0.225％FA)-ACN]；B％：12％-42％，10分钟)直接纯化，得到化合物11(24mg,50.2μmol，收率：42.9％)为浅黄色固体。MS(ESI):479.5[M+1] +。
1H NMR(400MHz,DMSO-d 6):δ12.34(s,1H),9.14(d,J＝2.0Hz,1H),8.66(d,J＝4.8Hz,1H),8.43(s,1H),8.18(dd,J＝2.2,8.2Hz,1H),7.98-7.93(m,1H),7.89(d,J＝1.6Hz,1H),7.70(d,J＝7.6Hz,1H),7.51(d,J＝3.2Hz,1H),7.37(dd,J＝4.9,7.1Hz,1H),7.13(s,1H),7.08(d,J＝7.6Hz,1H),6.64(d,J＝3.2Hz,1H),6.39(d,J＝8.2Hz,1H),5.69(s,2H),1.61(s,6H),0.49(s,2H),0.34-0.33(m,2H)。
实施例12
3-(1-(1-([3,3'-联吡啶]-6-基甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸12
Figure PCTCN2020072487-appb-000038
按照与实施例11类似的制备方法，在步骤1中由吡啶-3-基硼酸替换化合物11-0，得到化合物12，浅黄色固体。MS(ESI):479.5。
1H NMR(400MHz,DMSO-d 6):δ12.21(s,1H),8.85(dd,J＝11.25,1.83Hz,2H),8.60(dd,J＝4.71,1.53Hz,1H),8.46-8.48(m,1H),8.08(dt,J＝8.07,1.90Hz,1H),7.93(dd,J＝8.13,2.38Hz,1H),7.72(dd,J＝7.82,1.10Hz,1H),7.49-7.53(m,2H),7.14-7.18(m,1H),7.07-7.12(m,1H),6.66(d,J＝3.18Hz,1H),6.39(d,J＝8.19Hz,1H),5.73(s,2H),1.63(s,6H),0.51-0.56(m,2H),0.33-0.39(m,2H)。
实施例13
3-(1-(1-([3,4'-联吡啶]-6-基甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸13
Figure PCTCN2020072487-appb-000039
按照与实施例11类似的制备方法，在步骤1中由吡啶-4-基硼酸替换化合物11-0，得到化合物13，浅黄色固体。MS(ESI):479.5。
1H NMR(400MHz,DMSO-d 6):δ12.20(s,1H),8.90(d,J＝2.0Hz,1H),8.67-8.63(m,2H),8.47(s,1H),7.99(dd,J＝2.4,8.3Hz,1H),7.74-7.68(m,3H),7.52(d,J＝3.2Hz,1H),7.19-7.14(m,1H),7.12-7.07(m,1H),6.66(d,J＝3.2Hz,1H),6.40(d,J＝8.3Hz,1H),5.74(s,2H),1.65(s,6H),0.57-0.52(m,2H),0.39-0.33(m,2H)。
实施例14
3-(1-(1-((5-(1-甲基-1H-吡唑-4-基)吡啶-2-基)甲基)-1H-吲哚-7-甲酰氨基)环丙基)双环[1.1.1]戊烷-1-羧酸14
Figure PCTCN2020072487-appb-000040
按照与实施例11类似的制备方法，在步骤1中由(1-甲基-1H-吡唑-4-基)硼酸替换化合物11-0，得到化合物14，白色固体。MS(ESI):482.2。
1H NMR(400MHz,DMSO-d 6):δ12.20(s,1H),8.69(d,J＝1.71Hz,1H),8.50(s,1H),8.15(s,1H),7.85(d,J＝0.61Hz,1H),7.68-7.72(m,2H),7.48(d,J＝3.18Hz,1H),7.14-7.16(m,1H),7.06-7.10(m,1H),6.63(d,J＝3.18Hz,1H),6.28(d,J＝8.19Hz,1H),5.62(s,2H),3.86(s,3H),1.69(s,6H),0.54-0.57(m,2H),0.39-0.43(m,2H)。
实施例15
3-(1-(1-((5-(1H-吡唑-4-基)吡啶-2-基)甲基)-1H-吲哚-7-甲酰氨基)环丙基)双环[1.1.1]戊烷-1-羧酸15
Figure PCTCN2020072487-appb-000041
按照与实施例11类似的制备方法，在步骤1中由(1H-吡唑-4-基)硼酸替换化合物11-0，得到化合物15，浅黄色固体。MS(ESI):467.2。
1H NMR(400MHz,DMSO-d 6):δ12.30(s,1H),8.74(d,J＝1.59Hz,1H),8.51-8.47(m,1H),8.32-7.86(m,2H),7.72(dd,J＝17.03,8.04,1.71Hz,2H),7.48(d,J＝3.18Hz,1H),7.16-7.13(m,1H),7.10-7.05(m,1H),6.63(d,J＝3.18Hz,1H),6.29(d, J＝8.19Hz,1H),5.62(s,2H),1.68(s,6H),0.56-0.53(m,2H),0.43-0.40(m,2H)。
实施例16
3-(1-(1-((5-(1-(3-羟基丙基)-1H-吡唑-4-基)吡啶-2-基)甲基)-1H-吲哚-7-甲酰氨基)环丙基)双环[1.1.1]戊烷-1-羧酸16
Figure PCTCN2020072487-appb-000042
按照与实施例11类似的制备方法，在步骤1中由3-(4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-1H-吡唑-1-基)丙-1-醇替换化合物11-0，得到化合物16，白色固体。MS(ESI):526.2。
1H NMR(400MHz,DMSO-d 6):δ12.20(s,1H),8.70(d,J＝2.0Hz,1H),8.46(s,1H),8.20(s,1H),7.86(s,1H),7.72-7.67(m,2H),7.48(d,J＝3.1Hz,1H),7.13(s,1H),7.08(d,J＝7.6Hz,1H),6.62(d,J＝2.9Hz,1H),6.29(d,J＝8.1Hz,1H),5.62(s,2H),4.16(t,J＝7.0Hz,2H),3.39(s,2H),1.95-1.89(m,2H),1.64(s,6H),1.24(s,1H),0.57-0.51(m,2H),0.42(d,J＝1.2Hz,2H)。
实施例17
3-(1-(1-((5-(3-羟基哌啶-1-基)吡啶-2-基)甲基)-1H-吲哚-7-甲酰氨基)环丙基)双环[1.1.1]戊烷-1-羧酸17
Figure PCTCN2020072487-appb-000043
按照与实施例11类似的制备方法，在步骤1中由哌啶-3-醇替换化合物11-0，在DMSO中，加入Cs 2CO 3(787mg,2.42mmol),2-(二甲基氨基)乙酸(25mg,242umol)and碘化亚铜(70.0mg,367umol)，反应在氮气下127℃进行13小时完成。其它反应步骤相同，得到化合物17，浅黄色固体。MS(ESI):501.1。
1H NMR(400MHz,DMSO-d 6):δ12.20(s,1H),8.46(s,1H),8.13(d,J＝2.29Hz,1H),7.66(d,J＝7.32Hz,1H),7.40(d,J＝3.20Hz,1H),7.12(d,J＝7.32Hz,1H),7.04-7.09(m,2H),6.57(d,J＝3.02Hz,1H),6.29(d,J＝8.51Hz,1H),5.47(s,2H),5.32(t,J＝4.89Hz,1H),3.49-3.60(m,4H),3.17(d,J＝5.03Hz,1H),2.00(d,J＝7.32Hz,2H),1.76(s,1H),1.64(s,6H),1.46(d,J＝2.20Hz,1H),0.55(s,2H),0.41-0.46(m,2H)。
实施例18
3-(1-(1-((5-环丙基吡啶-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸18
Figure PCTCN2020072487-appb-000044
按照与实施例11类似的制备方法，在步骤1中由环丙基硼酸替换化合物11-0，得到化合物18，白色固体。MS(ESI):442.2。
1H NMR(400MHz,DMSO-d 6):δ12.20(s,1H),δ8.43(s,1H),8.28(d,J＝2.3Hz,1H),7.68(dd,J＝1.2,7.8Hz,1H),7.44(d,J＝3.2Hz,1H),7.20-7.11(m,2H),7.09-7.03(m,1H),6.60(d,J＝3.2Hz,1H),6.20(d,J＝7.7Hz,1H),5.58(s,2H),1.88(s,1H),1.64(s,6H),0.97-0.89(m,2H),0.66-0.61(m,2H),0.59-0.54(m,2H),0.40-0.31(m,2H)。
实施例19
3-(1-(1-((5-(2-(羟基甲基)苯基)吡啶-2-基)甲基)-1H-吲哚-7-甲酰氨基)环丙基)双环[1.1.1]戊烷-1-羧酸19
Figure PCTCN2020072487-appb-000045
按照与实施例11类似的制备方法，在步骤1中由(2-(羟甲基)苯基)硼酸替换化合物11-0，得到化合物19，白色固体。MS(ESI):508.2。
1H NMR(400MHz,DMSO-d 6):δ12.20(s,1H),8.53(s,1H),8.48(d,J＝1.75Hz,1H),7.73(dd,J＝7.88,1.00Hz,1H),7.61(dd,J＝8.07,2.19Hz,1H),7.53-7.58(m,2H),7.41(td,J＝7.47,1.19Hz,1H),7.34(td,J＝7.47,1.19Hz,1H),7.19(s,1H),7.17(s,1H),7.08-7.12(m,1H),6.66(d,J＝3.13Hz,1H),6.29(d,J＝8.00Hz,1H),5.74(s,2H),4.34(s,2H),1.72(s,6H),0.54-0.59(m,2H),0.32-0.37(m,2H)。
实施例20
3-(1-(1-(苯并呋喃-2-基甲基)-1H-吲哚-7-甲酰胺基)乙基)双环[1.1.1]戊烷-1-羧酸20、(S)-3-(1-(1-(苯并呋喃-2-基甲基)-1H-吲哚-7-甲酰胺基)乙基)双环[1.1.1]戊烷-1-羧酸20-5和(R)-3-(1-(1-(苯并呋喃-2-基甲基)-1H-吲哚-7-甲酰胺基)乙基)双环[1.1.1]戊烷-1-羧酸20-6
Figure PCTCN2020072487-appb-000046
步骤1：3-(氯羰基)双环[1.1.1]戊烷-1-羧酸甲酯20-1的制备
将3-(甲氧基羰基)双环[1.1.1]戊烷-1-羧酸(100mg，0.59mmol)的SOCl 2(2mL)溶液在80℃下搅拌1.5小时。浓缩反应混合物，得到化合物20-1(110mg，收率：100％)，为白色固体，将其不经纯化用于下一步骤。
步骤2：3-乙酰基双环[1.1.1]戊烷-1-羧酸甲酯20-2的制备
在0℃，氮气氛下，向CuI(134mg，0.71mmol)的无水THF(2mL)悬浮液中滴加MeLi(0.88mL，1.6M的THF溶液，1.4mmol)。然后将反应混合物冷却至-78℃并滴加化合物20-1(110mg，0.59mmol)的无水THF(2mL)溶液。将反应混合物在-78℃下搅拌2小时。然后加入甲醇(0.6mL)并将混合物升温至室温。加入饱和NH 4Cl溶液(10mL)，用EtOAc(20mL×3)萃取混合物。将合并的有机层用水和盐水洗涤，用无水Na 2SO 4干燥，过滤并浓缩。通过柱色谱(硅胶，用己烷/EtOAc从5：1至2：1(v/v)洗脱)纯化残余物，得到化合物20-2(75mg，产率：76％)，为浅黄色。固体。MS(ESI):169.1[M+1] +。
1H NMR(400MHz,CDCl3)δ:3.62(s,3H),2.24(s,6H),2.07(s,3H)。
步骤3：3-(1-氨基乙基)双环[1.1.1]戊烷-1-羧酸甲酯20-3的制备
将化合物20-2(293mg，1.74mmol)，NH 4OAc(805mg，10.46mmol)和NaBH 3CN(164mg，2.61mmol)在MeOH(6mL)中的混合物在室温下搅拌2小时。在通过LCMS指示反应完成后，浓缩反应混合物并通过柱色谱法(硅胶，用DCM/MeOH从30:1(v/v)至10：1(v/v)洗脱)纯化，得到化合物20-3(800mg，粗品，产率：100％)，为透明油状物。MS(ESI):170.1[M+1] +。
步骤4：3-(1-(1-(苯并呋喃-2-基甲基)-1H-吲哚-7-甲酰胺基)乙基)双环[1.1.1]戊 烷-1-羧酸甲酯20-4的制备
向化合物20-3(150mg，0.51mmol)，1-(苯并呋喃-2-基甲基)-1H-吲哚-7-羧酸(800mg，0.51mmol)和HATU(232mg，0.61mmol)的DMF(6mL)溶液中加入DIPEA(197mg，1.53mmol)。将所得混合物在室温下搅拌过夜。在通过LCMS 指示反应完成后，将反应混合物倒入水(5mL)中并用乙酸乙酯(15mL×3)萃取。将合并的有机层用盐水洗涤，用Na 2SO 4干燥，过滤并在真空下浓缩，得到残余物，将其通过柱色谱(硅胶，用PE/EtOAc从5：1至1：2洗脱(v/v)纯化。得到化合物20-4(117mg，收率：52％)，为浅黄色固体。MS(ESI):443.0[M+1] +。
步骤5：3-(1-(1-(苯并呋喃-2-基甲基)-1H-吲哚-7-甲酰胺基)乙基)双环[1.1.1]戊 烷-1-羧酸20的制备
向化合物20-4(106mg，0.24mmol)的MeOH(4mL)溶液中加入LiOH·H 2O(2M，0.15mL，0.30mmol)，并将所得混合物在50℃搅拌过夜。在通过LCMS指示反应完成后，用1M HCl将其酸化至pH～5并用乙酸乙酯(15mL×3)萃取。将合并的有机层用盐水洗涤，用Na 2SO 4干燥，过滤并在真空下浓缩，得到化合物20(80mg，收率：78％)，为灰白色固体。MS(ESI):429.0[M+1] +。
1H NMR(400MHz,DMSO-d 6)δ:12.29(brs,1H),8.32(d,J＝6.0Hz,1H),7.70(d,J＝7.2Hz,1H),7.57(d,J＝3.2Hz,1H),7.50(d,J＝8.0Hz,1H),7.44(d,J＝8.0Hz,1H),7.22-7.09(m,4H),6.61(d,J＝2.8Hz,1H),6.39(s,1H),5.91(d,J＝16.0Hz,1H),5.68(d,J＝16.0Hz,1H),4.18-4.11(m,1H),1.82(s,6H),0.93(d,J＝6.8Hz,3H)。
步骤6:(R)-3-(1-(1-(苯并呋喃-2-基甲基)-1H-吲哚-7-甲酰胺基)乙基)双环[1.1.1] 戊烷-1-羧酸20-5和(S)-3-(1-(1-(苯并呋喃-2-基甲基)-1H-吲哚-7-甲酰胺基)乙基)双 环[1.1.1]戊烷-1-羧酸20-6的制备
通过手性HPLC使用CHIRALPAK AD-3(4.6×100mm,3um分离外消旋体20(80mg，0.19mmol)，其中MeOH为流动相，柱温35℃,得化合物20-5(20mg，Rt＝1.68min,收率：50％)和20-6(20mg，Rt＝3.09min,收率：50％),均为白色固体。
化合物20-5,MS(ESI):429.0[M+1] +。
1H NMR(400MHz,DMSO-d 6)δ:12.29(brs,1H),8.34(d,J＝6.0Hz,1H),7.70(d,J＝7.2Hz,1H),7.57(d,J＝3.2Hz,1H),7.50(d,J＝8.0Hz,1H),7.44(d,J＝8.0Hz,1H),7.22-7.06(m,4H),6.61(d,J＝2.8Hz,1H),6.39(s,1H),5.92(d,J＝16.0Hz,1H),5.68(d,J＝16.0Hz,1H),4.18-4.11(m,1H),1.82(s,6H),0.93(d,J＝6.8Hz,3H)。
化合物20-6,MS(ESI):429.0[M+1] +。
1H NMR(400MHz,DMSO-d 6)δ:12.46(brs,1H),8.34(d,J＝8.0Hz,1H),7.69(d,J＝8.0Hz,1H),7.57(d,J＝3.2Hz,1H),7.50(d,J＝6.8Hz,1H),7.44(d,J＝8.0Hz,1H),7.22-7.05(m,4H),6.61(d,J＝3.2Hz,1H),6.39(s,1H),5.92(d,J＝16.4Hz,1H),5.68(d,J＝16.4Hz,1H),4.18-4.11(m,1H),1.83(s,6H),0.93(d,J＝6.8Hz,3H)。.
实施例21
3-(1-(1-((5-氟苯并呋喃-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸21
Figure PCTCN2020072487-appb-000047
步骤1：5-氟苯并呋喃-2-羧酸甲酯21-1的合成
在反应瓶中，依次加入5-氟-2-羟基苯甲醛21-0(3.0g，21.41mmol)，溴乙酸甲酯(4.91g，32.12mmol)，碳酸钾(5.91g，42.82mmol)和N,N-二甲基甲酰胺(120mL)，在氮气保护下，加热到80℃搅拌过夜。反应完毕后，将反应液倒入水中，乙酸乙酯(200mL x 3)萃取，有机相依次用水和饱和食盐水水洗，无水硫酸钠干燥，减压蒸除溶剂，得粗品，经硅胶柱[洗脱剂：石油醚-乙酸乙酯(20:1-5:1)]纯化，得化合物21-1(2.1g，收率：51.8％)为黄色固体。MS(ESI):195.2[M+1] +。
步骤2：(5-氟苯并呋喃-2-基)甲醇21-2的合成
在反应瓶中，加入化合物21-1(2.1g，10.82mmol)和四氢呋喃(50mL)，在氮气保护下搅拌,冰浴冷却至0℃。将四氢铝锂(2.05g，54.10mmol)分批慢慢加入其中,保持温度在0℃。加料完毕,继续在0℃下搅拌1小时。依次将水(2mL),15％氢氧化钠水溶液(2mL)和水(4mL)慢慢滴加到反应液中，保持温度在0℃,滴加完毕,升至室温，继续搅拌1小时。过滤，收集滤液，无水硫酸钠干燥，得化合物21-2(1.71g，收率：95.1％)为无色油状物。MS(ESI):149.2[M+1] +。
步骤3：2-(溴甲基)-5-氟苯并呋喃21-3的合成
在反应瓶中，加入化合物21-2(1.0g，6.02mmol)和干燥的二氯甲烷(20mL)，在搅拌下冰浴冷却至0℃。将三溴化磷(1.8g，6.62mmol)缓慢滴加其中，加料完毕后，室温搅拌1小时。经TLC监测(石油醚：乙酸乙酯＝5：1)反应完毕后，减压蒸除溶剂，然后用饱和碳酸氢钠水溶液调pH至8-9，二氯甲烷(20mL x 3)萃取，合并的有机相用饱和食盐水洗，无水硫酸钠干燥，减压蒸除溶剂，得化合物21-3(1.3g，收率：94.3％)为黄色固体。MS(ESI):228.0[M+1] +。
步骤4-7：
按照实施例1中描述的方法，由化合物21-3替换1-(溴甲基)-4-(三氟甲基)苯，得到化合物21为灰白色固体。MS(ESI):459.1[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ12.24(s,1H),8.72(s,1H),7.77(d,J＝8.0Hz,1H),7.46-7.52(m,2H),7.22-7.29(m,2H),7.02-7.09(m,2H),6.60(d,J＝3.2Hz,1H),6.21(s,1H),5.75(s,2H),1.69(s,6H),0.59-0.62(m,2H),0.45-0.48(m,2H)。
实施例22
3-(1-(1-((6-氟苯并呋喃-2-基)甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸22
Figure PCTCN2020072487-appb-000048
按照实施例21中描述的方法，由4-氟-2-羟基苯甲醛替换化合物21-0，获得化合物22，为灰白色固体。MS(ESI):459.1[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ12.24(s,1H),8.72(s,1H),7.69(d,J＝8.0Hz,1H),7.46-7.67(m,2H),7.39-7.42(d,J＝8.0,1H),7.22–7.24(d,J＝7.9,1H),7.02-7.09(m,2H),7.03(s,1H),6.64(d,J＝3.2Hz,1H),6.6(s,1H),5.75(s,2H),1.69(s,6H),0.64-0.62(m,2H),0.44-0.46(m,2H)。
实施例23
3-(1-(1-(苯并[d]噁唑-2-基甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸23
Figure PCTCN2020072487-appb-000049
按照实施例21中描述的方法，由苯并[d]噁唑-2-羧酸甲酯替换化合物21-0，得到化合物23，为白色固体。MS(ESI):442.2[M+1] +。
1H NMR(400MHz,DMSO-d6)δ:12.20(s,1H),8.57(s,1H),7.87(d,J＝7.6Hz,2H),7.69-7.6(m,2H),7.56-7.52(m,2H),7.34-7.18(m,2H),6.67(s,1H),6.23(s,2H),1.72(s,6H),0.56-0.47(m,4H)。
实施例24
3-(1-(1-(苯并[d]噻唑-2-基甲基)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸24
Figure PCTCN2020072487-appb-000050
按照实施例21中描述的方法，由苯并[d]噻唑-2-羧酸甲酯替换化合物21-0，得到化合物24，为白色固体。MS(ESI):458.2[M+1] +。
1H NMR(400MHz,DMSO-d6)δ:12.20(s,1H),8.57(s,1H),7.87(d,J＝7.6Hz,2H),7.69-7.6(m,2H),7.46-7.42(m,2H),7.34-7.18(m,2H),6.67(s,1H),6.03(s,2H),1.72(s,6H),0.56-0.47(m,4H)。
实施例25
3-(1-(1-(苯并呋喃-2-基甲基)-1H-吡咯并[3,2-c]吡啶-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸25
Figure PCTCN2020072487-appb-000051
步骤1：1-(苯并呋喃-2-基甲基)-7-溴-1H-吡咯并[3,2-c]吡啶25-1的合成
在0℃下，向7-溴-1H-吡咯并[3,2-c]吡啶25-0(1.79g，9.09mmol)在DMF(80mL)中的溶液中加入t-BuOK(1.53g，13.63mmol)，然后加入化合物6-2(2.5g，粗品，11.36mmol)。将所得混合物在室温搅拌过夜。然后将反应混合物倒入水(200ml)中，并用乙酸乙酯(100ml×2)萃取。合并的有机层用盐水洗涤，经Na 2SO 4干燥，过滤并在真空下浓缩，得到粗产物，将其通过柱色谱法纯化(石油醚/乙酸乙酯＝10/1-5/1)，得到化合物25-1(2.5g，产率：84％)为黄色油。MS(ESI):327.0[M+1] +。
步骤2：1-(苯并呋喃-2-基甲基)-1H-吡咯并[3,2-c]吡啶-7-羧酸甲酯25-2的合成
向化合物25-1(2.5克，7.64mmol)的MeOH(21ml)和DMF(14ml)溶液中添加Pd(OAc) 2(171mg，0.764mmol)，Ph 2P(CH 2) 3PPh 2(318毫克，0.764mmol)和TEA(2.31g，22.9mmol)。将得到的混合物在CO气氛下在80℃搅拌48小时。然后将反应混合物倒入水(100ml)中，并用乙酸乙酯(100ml×2)萃取。合并的有机层用盐水洗涤，经Na 2SO 4干燥，过滤并在真空下浓缩，得到粗产物，将其通 过柱色谱法纯化
Figure PCTCN2020072487-appb-000052
得到化合物25-2(1.3克，产率：55％)为黄色油。MS(ESI):307.2[M+1] +。
1H NMR(400MHz,DMSO-d 6)δ:9.02(s,1H),8.58(s,1H),7.95(s,1H),7.77(d,J＝3.6Hz,1H),7.56(dd,J＝7.2,0.8Hz,1H),7.44-7.42(m,1H),7.24-7.18(m,2H),6.85(d,J＝3.6Hz,1H),6.56(d,J＝0.8Hz,1H),5.89(s,2H),3.91(s,3H)。
步骤3：1-(苯并呋喃-2-基甲基)-1H-吡咯并[3,2-c]吡啶-7-羧酸25-3的合成
向化合物25-2(2g，1.96mol)的THF(10mL)和甲醇(10mL)溶液中加入NaOH(2M，4.9mL，9.8mmol)的水溶液。将所得混合物在室温搅拌过夜。然后将反应混合物浓缩以除去MeOH和THF，得到残余物，将其倒入水(30.0mL)中，并用1N盐酸水溶液酸化至
Figure PCTCN2020072487-appb-000053
过滤收集形成的沉淀，得到化合物25-3(350mg，产率：61％)，为白色固体。MS(ESI):293.0[M+1] +。
1H NMR(400MHz,DMSO-d 6)δ:13.60(brs,1H),9.02(s,1H),8.61(s,1H),7.77(d,J＝3.2Hz,1H),7.54(dd,J＝7.6,0.8Hz,1H),7.43(t,J＝7.6Hz,1H),7.26-7.16(m,2H),6.86(d,J＝3.2Hz,1H),6.50(s,1H),6.01(s,2H)。
步骤4：3-(1-(1-(1-(苯并呋喃-2-基甲基)-1H-吡咯并[3,2-c]吡啶-7-甲酰胺基)环 丙基)双环[1.1.1]戊烷-1-羧酸甲酯25-4的合成
向化合物25-3(60mg，0.2mmol)，化合物A-1(41mg，0.22mmol)和HATU(114mg，0.3mmol)的DMF(5mL)溶液中加入DIPEA(77mg，0.6)。所得混合物在室温下搅拌过夜。将反应混合物倒入水(10mL)中，并用乙酸乙酯(10mL×3)萃取。合并的有机层用盐水洗涤，经Na 2SO 4干燥，过滤并在真空下浓缩，得到残余物，将其通过柱色谱法纯化
Figure PCTCN2020072487-appb-000054
得到化合物25-4(60毫克，产率：64％)为黄色油状液体。MS(ESI):456.0[M+1] +。
步骤5：3-(1-(1-(苯并呋喃-2-基甲基)-1H-吡咯并[3,2-c]吡啶-7-甲酰胺基)环丙基) 双环[1.1.1]戊烷-1-羧酸25的合成
向化合物25-4(60mg，0.13mmol)的MeOH(3mL)溶液中加入LiOH·H 2O(2M，0.33mL，0.66mmol)的水溶液，所得混合物在50℃搅拌过夜。然后将反应混合物浓缩以除去MeOH，得到残余物，将其用1M盐酸水溶液酸化至
Figure PCTCN2020072487-appb-000055
过滤收集形成的沉淀，得到化合物25(30mg，产率：51％)，为白色固体。MS(ESI):441.1[M+1] +。MS(ESI):442.2[M+1] +。
1H NMR(400MHz,DMSO-d 6)δ:12.25(s,1H),8.99(s,1H),8.92(s,1H),8.30(s,1H),7.64(d,J＝3.6Hz,1H),7.49(d,J＝7.2Hz,1H),7.43(d,J＝8.0Hz,1H),7.25-7.15(m,2H),6.78(d,J＝3.2Hz,1H),6.31(s,1H),5.82(s,2H),1.76(s,6H),0.64(t,J＝7.6Hz,2H),0.53(t,J＝1.6Hz,2H)。
实施例26
3-(1-(1-(苯并呋喃-2-基甲基)-1H-吡咯并[3,2-c]吡啶-7-甲酰胺基)乙基)双环[1.1.1]戊 烷-1-羧酸26、(S)-3-(1-(1-(苯并呋喃-2-基甲基)-1H-吡咯并[3,2-c]吡啶-7-甲酰胺基)乙基)双环[1.1.1]戊烷-1-羧酸26-1、(R)-3-(1-(1-(苯并呋喃-2-基甲基)-1H-吡咯并[3,2-c]吡啶-7-甲酰胺基)乙基)双环[1.1.1]戊烷-1-羧酸26-2
Figure PCTCN2020072487-appb-000056
按照实施例23中描述的方法，由化合物21-3替换化合物A1，获得化合物26，为灰白色固体。MS(ESI):430.1[M+1] +。
1H NMR(400MHz,DMSO-d 6)δ:12.29(s,1H),8.92(s,1H),8.56(d,J＝8.4Hz,1H),8.27(s,1H),7.69(d,J＝3.2Hz,1H),7.51(d,J＝7.2Hz,1H),7.43(d,J＝8.4Hz,1H),7.24-7.16(m,2H),6.78(d,J＝3.2Hz,1H),6.47(s,1H),5.92(d,J＝16.4Hz,1H),5.74(d,J＝16.4Hz,1H),4.15(t,J＝7.2Hz,1H),1.83(s,6H),0.94(d,J＝6.8Hz,3H)。
然后，通过手性HPLC使用CHIRALPAK AD-3(4.6×100mm,3um)分离外消旋体26(80mg，0.19mmol)，其中MeOH为流动相，得化合物26-1(Rt＝1.68min,20mg，收率50％和26-2(Rt＝3.09min,20mg，收率：50％)，均为白色固体。
化合物26-1，为白色固体。MS(ESI):430.1[M+1] +。
1H NMR(400MHz,DMSO-d 6)δ:12.29(s,1H),8.92(s,1H),8.56(d,J＝8.4Hz,1H),8.27(s,1H),7.69(d,J＝3.2Hz,1H),7.51(d,J＝7.2Hz,1H),7.43(d,J＝8.4Hz,1H),7.24-7.16(m,2H),6.78(d,J＝3.2Hz,1H),6.47(s,1H),5.92(d,J＝16.4Hz,1H),5.74(d,J＝16.4Hz,1H),4.15(t,J＝7.2Hz,1H),1.83(s,6H),0.94(d,J＝6.8Hz,3H).
化合物26-2，为白色固体。MS(ESI):430.1[M+1] +。
1H NMR(400MHz,DMSO-d 6)δ:12.29(s,1H),8.92(s,1H),8.56(d,J＝8.0Hz,1H),8.27(s,1H),7.69(d,J＝3.2Hz,1H),7.51(d,J＝7.2Hz,1H),7.43(d,J＝8.4Hz,1H),7.24-7.16(m,2H),6.78(d,J＝3.2Hz,1H),6.47(s,1H),5.92(d,J＝16.4Hz,1H),5.74(d,J＝16.4Hz,1H),4.15(t,J＝7.2Hz,1H),1.83(s,6H),0.94(d,J＝6.8Hz,3H)。
按照实施例23中描述的方法，用适当的化合物作为起始原料，获得以下实施例化合物：
Figure PCTCN2020072487-appb-000057
Figure PCTCN2020072487-appb-000058
实施例29
3-(1-(1-(苯并呋喃-2-基甲基-d)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸29
Figure PCTCN2020072487-appb-000059
步骤1：苯并呋喃-2-基甲烷-d-醇29-1的合成
在反应瓶中，依次加入化合物6-0(3克,20.5mmol)和无水甲醇(40ml)，冷却至0℃，将硼氢化钠-d 4(0.545g,14.4mmol)分批加入其中,保持温度低于25℃。加料完毕,在室温下搅拌1小时。反应完毕，减压蒸除溶剂，加入1N HCl水溶液(15ml)，室温搅拌5分钟，然后用饱和碳酸氢钠水溶液调pH至8-9，乙酸乙酯(10ml x 3)萃取，合并的有机相用饱和食盐水水洗，无水硫酸钠干燥，减压蒸除溶剂，得黄色油状的化合物29-1(3.0g)。MS(ESI):150.1[M+1] +。
步骤2：2-(溴甲基-d)苯并呋喃29-2的合成
在反应瓶中，依次加入化合物29-1(2.47g，16.7mmol)和干燥的二氯甲烷(32ml)，冷却至0℃。将三溴化磷(1.72mL，18.4mmol)缓慢滴加其中，加料完毕，升温至室温搅拌1小时。经TLC监测(石油醚：乙酸乙酯＝9：1)反应完毕，用饱和碳酸氢钠水溶液调pH至8-9，二氯甲烷(10ml x 3)萃取，合并的有机相用饱和食盐水水洗，无水硫酸钠干燥，减压蒸除溶剂，得黄色油状的化合物29-2(3.31g，收率：95％)。直接用于下一步的反应。
然后，按照实施例6中描述的方法，由化合物29-2替换化合物6-2，得到化合物29，为白色固体。MS(ESI):442.1[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ12.24(s,1H),8.75(s,1H),7.77(d,J＝8.0Hz,1H),7.43-7.30(m,2H),7.28-7.21(m,2H),7.17(d,J＝6.8Hz,1H),7.14-7.09(m, 1H),6.64(d,J＝3.2Hz,1H),6.21(s,1H),6.06(s,1H),5.95(s,1H),1.76(s,6H),0.64-0.62(m,2H),0.52-0.49(m,2H)。
实施例30
3-(1-(1-(1-苯并呋喃-2-基甲基-d2)-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸30
Figure PCTCN2020072487-appb-000060
步骤1：苯并呋喃-2-基甲烷-d2-醇30-1的合成
在反应瓶中，加入苯并呋喃-2-羧酸甲酯30-0(1.9g，10.8mmol)和四氢呋喃(50mL)，在氮气保护下搅拌,冰浴冷却至0℃。将四氢铝锂-d4(2.2g，54.10mmol)分批慢慢加入其中,保持温度在0℃。加料完毕,继续在0℃下搅拌1小时。依次将水(2mL),15％氢氧化钠水溶液(2mL)和水(4mL)慢慢滴加到反应液中，保持温度在0℃,滴加完毕,升至室温，继续搅拌1小时。过滤，收集滤液，无水硫酸钠干燥，得化合物30-1(1.48g,收率：91％)，为无色油状物。MS(ESI):151.2[M+1] +。
然后，按照实施例6中描述的方法，由化合物30-1替换化合物6-1，得到化合物30，为白色固体。MS(ESI):443.1[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ12.24(s,1H),8.75(s,1H),7.77(d,J＝8.0Hz,1H),7.43-7.30(m,2H),7.28-7.21(m,2H),7.17(d,J＝6.8Hz,1H),7.14-7.09(m,1H),6.64(d,J＝3.2Hz,1H),6.21(s,1H),6.06(s,1H),1.76(s,6H),0.64-0.62(m,2H),0.52-0.49(m,2H)。
实施例31
3-(1-(1-(1-(苯并呋喃-2-基甲基)-1H-吲哚-7-甲酰胺基)环丙基-2,2,3,3-d4)双环[1.1.1]戊烷-1-羧酸31
Figure PCTCN2020072487-appb-000061
步骤1：氘代格氏试剂的合成
在装有回流管的干燥的圆底烧瓶中加入乙醚(50ml)、溴乙烷-d5(5.6g,50mmol)和镁条(1.32g,55mmol)。所得混合物在氮气下搅拌并在反应混合物中加入少量的碘加热至回流。当多数的的镁条都消耗完后，反应停止加热。所得氘代格氏试剂31-1用作下一步反应之用。
步骤2：3-(1-氨基环丙基-2,2,3,3-d4)双环[1.1.1]戊烷-1-羧酸甲酯的合成
在氮气保护及-20℃的温度下，向3-氰基双环[1.1.1]戊烷-1-羧酸甲酯(3.0g，20mmol)的甲苯(50mL)溶液中加入四异丙氧基钛(5.9g，21mmol，6mL，95％纯度)。然后在氮气保护及-20℃的温度下，在30分钟内逐滴加入化合物31-1(1M，42mL,42mmol)，保持温度在-20～-10℃之间。搅拌30分钟后，逐滴加入BF 3·Et 2O(6g，42mmol，5.2mL)并在-20℃下搅拌30分钟，然后在25℃下搅拌12小时。TLC(板1：石油醚/乙酸乙酯＝3/1)显示原料(Rf＝0.61)已经消耗完毕，TLC(板2：石油醚/乙酸乙酯＝1/1)显示有一个主要产品(Rf＝0.24)形成。在0℃下缓慢加入盐酸水溶液(1N,30mL)将反应混合物淬灭，然后弃去分离的有机层。在0℃下用10M氢氧化钠水溶液将水相碱化至pH～12并用乙酸乙酯(200mL×2)萃取。将合并的有机层浓缩，得到残余物，将其通过柱色谱(二氧化硅，石油醚/乙酸乙酯＝20/1至1/1)纯化，得到3-(1-氨基环丙基-2,2,3,3-d4)双环[1.1.1]戊烷-1-羧酸甲酯(1.2g，6.4mmol，32％收率)，为黄色固体。MS(ESI):186.1[M+1] +。
然后，按照实施例6中描述的方法，由3-(1-氨基环丙基-2,2,3,3-d4)双环[1.1.1]戊烷-1-羧酸甲酯A1-d4替换化合物A1，得到化合物31，为灰白色固体。MS(ESI):445.1[M+1] +。
1H NMR(400MHz,DMSO-D 6):δ12.24(s,1H),8.75(s,1H),7.77(d,J＝8.0Hz,1H),7.43-7.30(m,2H),7.28-7.21(m,2H),7.17(d,J＝6.8Hz,1H),7.14-7.09(m,1H),6.64(d,J＝3.2Hz,1H),6.21(s,1H),6.06(s,1H),5.75(s,2H),1.76(s,6H)。
实施例32
3-(1-(1-(1-(苯并呋喃-2-基甲基)-5-氟-1H-吲哚-7-甲酰胺基)环丙基)双环[1.1.1]戊烷-1-羧酸32
Figure PCTCN2020072487-appb-000062
步骤1：1-(苯并呋喃-2-基甲基)-7-溴-5-氟-1H-吲哚32-1的合成
在反应瓶中，加入化合物6-2(251mg，1.17mmol)和N,N-二甲基甲酰胺(10mL)，在搅拌下冰浴冷却至0℃，随后依次加入叔丁醇钾(171mg，1.52mmol)和7-溴-5-氟-1H-吲哚32-0(295mg，1.40mmol)，加料完毕后，室温搅拌1小时。反应完毕后，将反应液倒入水中，乙酸乙酯(100mL x 3)萃取，有机相依次用水和饱和食盐水水洗，无水硫酸钠干燥，减压蒸除溶剂，得粗品，经硅胶柱[洗脱剂：石油醚-乙酸乙酯(20:1-10:1)]纯化，得化合物32-1(390mg，收率：96.5％)为黄色油状物。MS(ESI):344.1[M+1] +。
步骤2：1-(苯并呋喃-2-基甲基)-5-氟-1H-吲哚-7-羧酸甲酯32-2的合成
在反应瓶中，依次加入化合物32-1(390mg，1.14mmol)，醋酸钯(26mg，0.12mmol),1,3-双(二苯基膦)丙烷(47mg，0.12mmol)，甲醇(5mL)，N,N-二甲基甲酰胺(10mL)和三乙胺(344mg，3.41mmol)，用一氧化碳气球置换三次，在一氧化碳环境下90℃搅拌48小时。反应完毕后，将反应液倒入水中，乙酸乙酯(50mL x 3)萃取，有机相依次用水和饱和食盐水洗，无水硫酸钠干燥，减压蒸除溶剂，得粗品，经硅胶柱[洗脱剂：石油醚-乙酸乙酯(30:1-1:1)]纯化，得化合物32-2(173mg，收率：47.1％)为黄色油状物。MS(ESI):324.1[M+1] +。
步骤3：1-(苯并呋喃-2-基甲基)-5-氟-1H-吲哚-7-羧酸32-3的合成
在反应瓶中，依次加入化合物32-2(173mg，0.54mmol)，四氢呋喃(5mL)，甲醇(5mL)和2M NaOH(1.34mL，2.68mmol)，室温搅拌过夜。反应完毕后，减压蒸除溶剂后，加入水(5mL)，然后用1M HCl水溶液调pH至1-2，过滤，滤饼用水洗，得化合物32-3(156mg，收率：94.5％)为黄色固体。MS(ESI):310.0[M+1] +。
步骤4：3-(1-(1-(1-(苯并呋喃-2-基甲基)-5-氟-1H-吲哚-7-甲酰胺基)环丙基)双环 [1.1.1]戊烷-1-羧酸甲酯32-4的合成
在反应瓶中，依次加入化合物32-3(90mg，0.29mmol)，化合物A-1(63mg，0.35mmol)，HATU(133mg，0.35mmol),N,N-二甲基甲酰胺(4mL)和N,N- 二异丙基乙胺(75mg，0.58mmol)，室温搅拌过夜。反应完毕后，将反应液倒入水中，乙酸乙酯(10mL x 3)萃取，有机相依次用水和饱和食盐水洗，无水硫酸钠干燥，减压蒸除溶剂，得粗品，经硅胶柱[洗脱剂：石油醚-乙酸乙酯(10:1-2:1)]纯化，得化合物32-4(112mg，收率：81.8％)为黄色固体。MS(ESI):473.1[M+1] +。
步骤5：3-(1-(1-(1-(苯并呋喃-2-基甲基)-5-氟-1H-吲哚-7-甲酰胺基)环丙基)双环 [1.1.1]戊烷-1-羧酸32的合成
在反应瓶中，依次加入化合物32-4(112mg，0.24mmol)，甲醇(5mL)和2M LiOH水溶液(0.18mL，0.36mmol)，加热到50℃搅拌过夜。反应完毕后，减压蒸除溶剂后，加入水(4mL)，然后用1M HCl水溶液调pH至5-6，过滤，滤饼依次用水洗，干燥，得化合物32(15mg，收率：13.9％)，为白色固体。MS(ESI):459.1[M+1] +。
1H NMR(400MHz,DMSO-d6)δ:12.24(s,1H),8.83(s,1H),7.58(d,J＝3.2Hz,1H),7.49-7.41(m,3H),7.23-7.12(m,2H),7.05(dd,J＝12.0,7.2Hz,1H),6.59(d,J＝3.2Hz,1H),6.18(s,1H),5.72(s,2H),1.73(s,6H),0.61(q,J＝4.8Hz,2H),0.49(q,J＝5.6Hz,2H)。
生物活性检测
测试例1：MCF7 cAMP测定
a)将MCF7细胞(ATCC，6000个细胞/孔/40μl)接种到具有培养基(含有10％FBS和1X PS的DMEM)的384孔细胞培养板中，在37℃和5％CO 2下孵育过夜。
b)移除培养基，加入40μl/孔无血清培养基，在37℃和5％CO 2下孵育5小时。
c)改变为18μL/孔HBSS缓冲液(Hepes 20mM，0.1％BSA，500μM IBMX)。
d)稀释DMSO中的化合物，1/5稀释，9+0剂量。
e)将2μL d)中的化合物加入到98μLHBSS缓冲液中。
f)然后将e)中1μL/孔稀释的化合物加入细胞中，最终起始浓度为1μM，在37℃下孵育1小时。
g)用1μL/孔PGE2(10mM DMSO储液)刺激细胞，终浓度为10nM，DMSO终浓度为0.2％，在37℃下孵育30分钟。
h)孵育后，加入10μL/孔cAMP-d2和10μL/孔抗cAMP，RT 1h。
i)在Envision上读取HTRF信号(665nm/615nm)。
数据分析：
高控制：1μM E7046
低控制：DMSO
背景：30μL/孔裂解缓冲液+10μL/孔抗cAMP
分析数据：原始日期-背景
抑制％：100-100×(高控制-分析数据)/(高控制-低控制)
Figure PCTCN2020072487-appb-000063
结果表明，本发明化合物具有优良的抑制PGE2/EP4信号传导的活性。
测试例2：在肝细胞中的清除率测试
肝细胞种类：小鼠
产品目录：M00505
种属：ICR/CD-1
性别：雄性
来源：BioreoclmationIVT
操作步骤：
1.工作溶液的准备
a.在适当的溶剂(DMSO)中制备10mM试验化合物和阳性对照的储备溶液。
b.在单独的锥形管中，将198μL 50％乙腈/50％水和2μL 10mM溶液混合，配制成100μM的测试化合物和阳性对照。
2.肝细胞的制备
a.将孵育培养基和肝细胞解冻培养基置于37℃水浴中，并在使用前加热至少15分钟
b.从储存中转移一小瓶冷冻保存的肝细胞，确保小瓶保持在低温，直到解冻过程发生。通过将小瓶置于37℃水浴中并轻轻摇动小瓶2分钟来解冻细胞。解冻完成后，用70％乙醇喷雾小瓶，将小瓶转移到生物安全柜中。
c.使用大口径移液器吸头将肝细胞转移到含有解冻培养基的50mL锥形管中。将50mL锥形管置于离心机中并以100g旋转10分钟。完成旋转后，吸出解冻培养基并在足够的培养基中重悬肝细胞，产生～1.5×10 6个细胞/mL。
3.稳定性测定步骤
a.将198μL肝细胞移液到96孔未包被的平板的每个孔中。将平板置于培养箱中以使肝细胞升温10分钟。
b.将2μL的100μM测试化合物或阳性对照移液到96孔未包被的板的各孔中以开始反应。将培养皿放回培养箱中设定的时间点。
c.在0、15、30、60、90和120分钟的时间点以25μL等分试样转移孔内容物。然后将等分试样与6体积(150μL)含有内标IS(100nM阿普唑仑，200nM咖啡因和100nM甲苯磺丁脲)的乙腈混合以终止反应。涡旋5分钟。将样品在3220g下离心45分钟。将100μL上清液的等分试样用100μL超纯水稀释，并将该混合物用于LC/MS/MS分析。所有孵育均一式两份进行。
d.数据分析
所有计算均使用Microsoft Excel进行。峰面积由提取的离子色谱图确定。通过化合物消失百分比对时间曲线的回归分析确定母体化合物的体外半衰期(t1/2)
体外半衰期(t1/2)由斜率值k确定：
t1/2＝0.693/k
使用以下等式(重复测定的平均值)将体外t1/2(以分钟计)转化为体外的内在清除率(CLint，以μL/min/0.5×10 6细胞)：
CLint＝kV/N
V＝孵育体积(0.2mL)
N＝每孔肝细胞数(0.1×10 6个细胞)
Figure PCTCN2020072487-appb-000064
Figure PCTCN2020072487-appb-000065
结果表明，本发明化合物在肝细胞中具有优良的稳定性，从而具有优良的代谢稳定性。
测试例3：化合物6在小鼠体内对CT-26结肠直肠肿瘤的抑制作用
对于同种异体移植肿瘤功效研究：6周龄雌性BALB/c小鼠,每只皮下植入1×10 6CT26癌细胞)(国家实验细胞资源共享服务平台，资源编号3111CCCC)，当肿瘤达到约60-80mm 3时，携带肿瘤的小鼠被随机分配到载体或治疗组。化合物6以10、30或150mg/kg的量在0.5％MC中悬浮液口服(p.o.)给药，每天施用一次。对载体组的小鼠每天重复给与口服蒸馏水，其给予时间与化合物6相同。每周记录2-3次肿瘤体积和体重。结果见图1和图2。
从图1可以看出，在单药情况下，不同剂量的化合物6和E7046对CT-26肿瘤生长有明显的抑制作用，在同等剂量下，化合物6明显优于E7046。
图2显示化合物6和E7046对小鼠的体重没有明显的影响。
测试例4：化合物6和抗PD-1抗体单独和联合施用在小鼠体内对CT-26结肠直肠 肿瘤的抑制作用
对于同种异体移植肿瘤功效研究：6周龄雌性BALB/c小鼠,每只皮下植入1×10 6CT26癌细胞，当肿瘤达到约60-80mm 3时，携带肿瘤的小鼠被随机分配到载体或治疗组。化合物6在单独施用或者联合施用时以30或150mg/kg的量在0.5％MC中悬浮液口服(p.o.)给药，每天施用一次。PD-1在单独施用或者联合施用时，腹膜内给予抗PD-1抗体(Invivomab anti-mouse PD-1(CD279)，Abcam公司)，以5mg/kg的量给药，每4天一次，共3次(Q4D×3)。对载体组和PD-1单独给药的小鼠，每天重复给与口服蒸馏水，其给予时间与化合物6相同。每周记录2-3次肿瘤体积和体重。结果见图3和图4。
从图3可以看出，化合物6在单独施用或者和抗PD-1抗体联合施用时，在小鼠体内对CT-26肿瘤生长有很好的抑制作用。
图4显示化合物6在单独施用和抗PD1抗体联用时对小鼠的体重没有明显的影响。
测试例5：化合物6和抗PD-1抗体单独和联合用药在小鼠体内对EM-6乳腺 肿瘤的抑制作用
对于同种异体移植肿瘤功效研究：6周龄雌性BALB/c小鼠,每只皮下植入5×10 5EMT-6癌细胞，当肿瘤达到约60-80mm 3时，携带肿瘤的小鼠被随机分配到载体或治疗组。化合物6在单独施用或者联合施用时以30mg/kg的量在0.5％MC中悬浮液口服(p.o.)给药，每天施用一次。PD-1在单独施用或者联合施用时，腹膜内给予抗PD-1抗体(Invivomab anti-mouse PD-1(CD279)，Abcam公司)，在第一次给药时，以20mg/kg的量给药，以后每5天一次，每次以10mg/kg的量给药。对载体组和PD-1单独给药的小鼠，每天重复给与口服蒸馏水，其给予时间与化合物6相同。每周记录2-3次肿瘤体积和体重。结果见图5和图6。
从图5可以看出，化合物6在单独施用或者和抗PD-1抗体联合用药时，在小鼠体内对EMT-6乳腺肿瘤有很好的抑制作用。
图6显示化合物6在单独施用和抗PD1抗体联用时对小鼠的体重没有明显的影响。

Claims (15)

  1. 一种通式(I)所示的化合物：
    Figure PCTCN2020072487-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：
    M 1、M 2、M 3、M 5、M 6和M 7各自独立地为N原子或C-R 4；
    M 4选自N原子和C原子；
    环A选自C 6-10芳基、C 3-6环烷基、5-10元杂芳基和3-6元杂环基；
    X为
    Figure PCTCN2020072487-appb-100002
    Y选自一个键、C 1-4亚烷基、-CR 5R 6-、-O-、-OC 1-4亚烷基-、-NR 9C 1-4亚烷基-和-NR 9-，所述C 1-4亚烷基任选被选自H原子、D原子、卤素、羟基、氰基、氨基、硝基、C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、羟基C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基中的一个或多个取代基所取代；
    R 1和R 2相同或不同，且各自独立地选自H原子、D原子、氰基、C 1-4烷基、C 1-4烷氧基、C 3-6环烷基、3-6元杂环基、-C(O)OR 4、C(O)NR 7R 8、-COR 4、-S(O) mR 4、-NR 7R 8、C 6-10芳基和5-10元杂芳基，所述C 1-4烷基和C 1-4烷氧基各自独立地任选被选自H原子、D原子、卤素、羟基、氰基、氨基、硝基、C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、羟基C 1-4烷基、C 3-6环烷基、3-6元杂环基、-C(O)OR 4、C(O)NR 7R 8、-COR 4、-NR 4C(O)NR 7R 8、-OC(O)NR 7R 8、-NR 7C(O)OR 4、-S(O) mR 4、-NR 7R 8、C 6-10芳基和5-10元杂芳基中的一个或多个取代基所取代；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，其中所述C 3-6环烷基和3-6元杂环基各自独立地任选被选自D原子、卤素、羟基、氰基、氨基、硝基、-C(O)OR 4、C(O)NR 7R 8、-COR 4、-NR 4C(O)NR 7R 8、-OC(O)NR 7R 8、-NR 7C(O)OR 4、-S(O) mR 4、-NR 7R 8、C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、羟基C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基中的一个或多个取代基所取代；
    R 3相同或不同，且各自独立地选自H原子、D原子、卤素、羟基、氰基、氨 基、硝基、C 1-4烷基、C 1-4烷氧基、C 3-6环烷基、3-6元杂环基、C 6-10芳基、5-10元杂芳基、-C(O)R 4、-C(O)OR 4、-OC(O)NR 7R 8、-NR 7C(O)OR 4、-S(O) mR 4、-NR 7R 8和-C(O)NR 7R 8，其中所述C 1-4烷基、C 1-4烷氧基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基各自独立地任选被选自C 1-4烷基、羟基C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、卤素、羟基、氰基、氨基和硝基中的一个或多个取代基所取代；
    R 4选自H原子、D原子、卤素、羟基、氨基、C 1-4烷基、C 1-4烷氧基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基，其中所述C 1-4烷基、C 1-4烷氧基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基各自独立地任选被选自C 1-4烷基、羟基C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、卤素、羟基、氰基、氨基和硝基中的一个或多个取代基所取代；
    R 5和R 6各自独立地选自H原子、D原子、C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基，其中所述C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基各自独立地任选被选自C 1-4烷基、羟基C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、卤素、羟基、氰基、氨基和硝基中的一个或多个取代基所取代；或者R 5和R 6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，其中所述C 3-6环烷基或3-6元杂环基各自独立地任选被选自卤素、羟基、氰基、氨基、硝基、C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、羟基C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基中的一个或多个取代基所取代；
    R 7和R 8各自独立地选自H原子、D原子、C 1-4烷基、卤代C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基，其中所述C 1-4烷基、卤代C 1-4烷基、C 3-6环烷基、3-6元杂环基、C 6-10芳基和5-10元杂芳基各自独立地任选被选自C 1-4烷基、羟基C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、卤素、羟基、氰基、氨基和硝基中的一个或多个取代基所取代；
    R 9选自H原子、D原子和C 1-4烷基，其中所述C 1-4烷基任选被选自羟基C 1-4烷基、C 1-4烷氧基、卤代C 1-4烷基、卤代C 1-4烷氧基、卤素、羟基、氰基、氨基和硝基中的一个或多个取代基所取代；
    m为0、1或2；以及
    n为0、1、2、3或4。
  2. 根据权利要求1所述的通式(I)所示的化合物，其通式为(II)所示的化合物：
    Figure PCTCN2020072487-appb-100003
    其中M 1、M 3、M 5和M 6各自独立地为CH或C-卤素，M 2为C-R 4或N原子，M 7为N原子，M 4为C原子，环A、R 1、R 2、R 3、R 4、X、Y和n如权利要求1中所定义。
  3. 根据权利要求1或2所述的通式(I)所示的化合物，其中Y为C 1-4亚烷基，优选亚甲基。
  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R 1和R 2相同或不同，且各自独立地选自H原子、D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，优选环丙基。
  5. 根据权利要求1-3中任一项所述的通式(I)所示的化合物，其为通式(III)所示的化合物：
    Figure PCTCN2020072487-appb-100004
    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可药用的盐，
    其中：
    R 1和R 2相同或不同，且各自独立地选自H原子、D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，优选环丙基；
    M 2为N原子或C-R 4；
    R 4选自H原子、D原子和卤素；
    X、环A、R 3和n如权利要求1中所定义。
  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，其中环A选自苯基、吡啶基、喹啉基、苯并呋喃基、吗啉基、吡唑、环丙基、异噁唑、苯并噁唑和苯并噻唑。
  7. 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 3相同或不同，且各自独立地选自H原子、D原子、卤素、C 1-4烷基、氟代C 1-4烷基、苯基、羟基C 1-4烷基取代的苯基、吗啉基、吡啶基、吡唑基、C 1-4烷基取代的吡唑基、羟基C 1-4烷基取代的吡唑基、环丙基、异噁唑基、哌啶基和羟基取代的哌啶基。
  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，其选自：
    Figure PCTCN2020072487-appb-100005
    Figure PCTCN2020072487-appb-100006
  9. 一种制备根据权利要求1-8中任一项所述的通式(I)所示的化合物的方法，其包括：
    Figure PCTCN2020072487-appb-100007
    通式(I-A)的化合物水解得到通式(I-B)的化合物，所述通式(I-B)的化合物和通式(I-C)的化合物发生缩合反应得到通式(I-D)的化合物，所述通式(I-D)的化合物水解脱去R之后得到通式(I)的化合物；
    其中：
    R为C 1-4烷基；
    M 1至M 7、环A、X、Y、R 1、R 2、R 3和n如权利要求1中所定义。
  10. 一种药物组合物，其包含治疗有效量的根据权利要求1-8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。
  11. 根据权利要求10所述的药物组合物，其进一步包含抗体，所述抗体选自抗CTLA4抗体、抗PDL1抗体和抗PD1抗体。
  12. 根据权利要求11所述的药物组合物，其中抗体选自易普利姆玛单抗、Tremelimumab、派姆单抗、纳武单抗、Atezolizumab、Avelumab、德瓦鲁单抗、Pidilizumab、AMP-224、AMP-514、PDR001、Cemiplimab、BMS-936559、CK-301、特瑞普利单抗、信迪利单抗、卡瑞利株单抗、替雷利株单抗、KN035、GLS-010、GB226、CS1001、CS1003、BAT-1306、HX008、AK105、LZM009、HLX10、HLX20、KL-A167、F520、GR1405和MSB2311。
  13. 根据权利要求1-8中任一项所述的通式(I)所示的化合物或根据权利要求10-12中任一项所述的药物组合物在制备抑制PGE2/EP4信号传导的药物中的用途。
  14. 根据权利要求1-8中任一项所述的通式(I)所示的化合物或根据权利要求10-12中任一项所述的药物组合物在制备治疗癌症的药物中的用途，优选地，所述癌症选自乳腺癌、宫颈癌、结肠直肠癌、子宫内膜癌、胶质母细胞瘤、头颈癌、肾癌、肝癌、肺癌、髓母细胞瘤、卵巢癌、胰腺癌、前列腺癌、皮肤癌和尿道癌。
  15. 根据权利要求1-8中任一项所述的通式(I)所示的化合物或根据权利要求10-12中任一项所述的药物组合物在制备治疗急性或慢性疼痛、偏头痛、骨关节炎、类风湿性关节炎、痛风、粘液囊炎、强直性脊柱炎、原发性痛经、癌症或动脉硬化症的药物中的用途。
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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022102731A1 (ja) 2020-11-13 2022-05-19 小野薬品工業株式会社 Ｅｐ４拮抗薬と免疫チェックポイント阻害物質との併用によるがん治療
WO2022111222A1 (zh) * 2020-11-26 2022-06-02 安润医药科技（苏州）有限公司 作为前列腺素ep4受体拮抗剂的酰胺衍生物及其用途
CN114591137A (zh) * 2022-05-10 2022-06-07 上海赛默罗生物科技有限公司 螺桨烷类衍生物的合成方法
WO2022247881A1 (zh) * 2021-05-28 2022-12-01 凯复(苏州)生物医药有限公司 治疗肿瘤的联合疗法
WO2022257961A1 (zh) * 2021-06-08 2022-12-15 武汉人福创新药物研发中心有限公司 用于治疗ep2、ep4受体介导的疾病的苯并杂环化合物
WO2023025270A1 (zh) * 2021-08-27 2023-03-02 凯复(苏州)生物医药有限公司 一种吲哚类化合物的固体形式及其制备方法和用途
WO2023160569A1 (en) * 2022-02-24 2023-08-31 Guangdong Newopp Biopharmaceuticals Co., Ltd. Carboxamide compounds as pge2 receptor antagonists

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020251957A1 (en) * 2019-06-11 2020-12-17 Teon Therapeutics, Inc. Prostaglandin e2 receptor 4 antagonists and uses thereof
CN119013017A (zh) * 2022-02-15 2024-11-22 治纳辅医药科技有限公司 包含抗癌剂和对前列腺素e2受体具有抑制活性的新化合物的用于治疗癌症的药物组合物
GB202211232D0 (en) 2022-08-02 2022-09-14 Heptares Therapeutics Ltd Prostaglandin EP4 receptor agonist compounds

Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008104055A1 (en) 2007-02-26 2008-09-04 Merck Frosst Canada Ltd. Indole and indoline cyclopropyl amide derivatives as ep4 receptor antagonists
WO2009048558A1 (en) * 2007-10-11 2009-04-16 Merck & Co., Inc. Maxi-k channel blockers and methods of use
EP2172447A1 (en) * 2007-07-03 2010-04-07 Astellas Pharma Inc. Amide compound
WO2013096496A2 (en) * 2011-12-21 2013-06-27 Allergan, Inc. Compounds acting at multiple prostaglandin receptors giving a general anti-inflammatory response
CN104411684A (zh) * 2012-06-29 2015-03-11 伊莱利利公司 苯氧基乙基哌啶化合物
WO2015179615A1 (en) 2014-05-23 2015-11-26 Eisai R&D Management Co., Ltd Combination therapies for the treatment of cancer
CN105793236A (zh) * 2013-12-17 2016-07-20 伊莱利利公司 二甲基苯甲酸化合物
WO2017041323A1 (zh) 2015-09-09 2017-03-16 深圳市华星光电技术有限公司 一种偏光片
WO2017066633A1 (en) 2015-10-16 2017-04-20 Eisai R&D Management Co., Ltd. Ep4 antagonists
WO2018084230A1 (en) 2016-11-04 2018-05-11 Askat Inc. Use of ep4 receptor antagonists for the treatment of nash-associated liver cancer
WO2018195123A1 (en) * 2017-04-18 2018-10-25 Tempest Therapeutics, Inc. Bicyclic compounds and their use in the treatment of cancer

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2565191B1 (en) * 2008-05-14 2014-10-08 Astellas Pharma Inc. 4-(Indol-7-ylcarbonylaminomethyl)cyclohexanecarboxylic acid derivatives as EP4 receptor antagonists useful for the treatment of chronic renal failure or diabetic nephropathy

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008104055A1 (en) 2007-02-26 2008-09-04 Merck Frosst Canada Ltd. Indole and indoline cyclopropyl amide derivatives as ep4 receptor antagonists
EP2172447A1 (en) * 2007-07-03 2010-04-07 Astellas Pharma Inc. Amide compound
WO2009048558A1 (en) * 2007-10-11 2009-04-16 Merck & Co., Inc. Maxi-k channel blockers and methods of use
WO2013096496A2 (en) * 2011-12-21 2013-06-27 Allergan, Inc. Compounds acting at multiple prostaglandin receptors giving a general anti-inflammatory response
CN104411684A (zh) * 2012-06-29 2015-03-11 伊莱利利公司 苯氧基乙基哌啶化合物
CN105793236A (zh) * 2013-12-17 2016-07-20 伊莱利利公司 二甲基苯甲酸化合物
WO2015179615A1 (en) 2014-05-23 2015-11-26 Eisai R&D Management Co., Ltd Combination therapies for the treatment of cancer
WO2017041323A1 (zh) 2015-09-09 2017-03-16 深圳市华星光电技术有限公司 一种偏光片
WO2017066633A1 (en) 2015-10-16 2017-04-20 Eisai R&D Management Co., Ltd. Ep4 antagonists
WO2018084230A1 (en) 2016-11-04 2018-05-11 Askat Inc. Use of ep4 receptor antagonists for the treatment of nash-associated liver cancer
WO2018195123A1 (en) * 2017-04-18 2018-10-25 Tempest Therapeutics, Inc. Bicyclic compounds and their use in the treatment of cancer

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
DIANA I. ALBU ET AL., ONCOIMMUNOLOGY, vol. 6, no. 8, 2017, pages e1338239
See also references of EP3889134A4

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022102731A1 (ja) 2020-11-13 2022-05-19 小野薬品工業株式会社 Ｅｐ４拮抗薬と免疫チェックポイント阻害物質との併用によるがん治療
WO2022111222A1 (zh) * 2020-11-26 2022-06-02 安润医药科技（苏州）有限公司 作为前列腺素ep4受体拮抗剂的酰胺衍生物及其用途
WO2022247881A1 (zh) * 2021-05-28 2022-12-01 凯复(苏州)生物医药有限公司 治疗肿瘤的联合疗法
WO2022257961A1 (zh) * 2021-06-08 2022-12-15 武汉人福创新药物研发中心有限公司 用于治疗ep2、ep4受体介导的疾病的苯并杂环化合物
WO2023025270A1 (zh) * 2021-08-27 2023-03-02 凯复(苏州)生物医药有限公司 一种吲哚类化合物的固体形式及其制备方法和用途
JP2024532601A (ja) * 2021-08-27 2024-09-05 キーセラ（スーチョウ）バイオ－ファーマスーティカルズ カンパニー，リミテッド インドール化合物の固体形態、その調製方法及びその使用
AU2022334637B2 (en) * 2021-08-27 2026-02-19 Keythera (Suzhou) Bio-Pharmaceuticals Co., Limited Solid form of indole compound, preparation method therefor and use thereof
WO2023160569A1 (en) * 2022-02-24 2023-08-31 Guangdong Newopp Biopharmaceuticals Co., Ltd. Carboxamide compounds as pge2 receptor antagonists
CN114591137A (zh) * 2022-05-10 2022-06-07 上海赛默罗生物科技有限公司 螺桨烷类衍生物的合成方法

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EP3889134C0 (en) 2024-09-25
CN111989311A (zh) 2020-11-24
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AU2020212111A1 (en) 2021-08-05
CA3126484A1 (en) 2020-07-30
TW202041498A (zh) 2020-11-16
KR102861190B1 (ko) 2025-09-17
KR20210117280A (ko) 2021-09-28
US12240812B2 (en) 2025-03-04
EP3889134A4 (en) 2022-02-16
JP7488269B2 (ja) 2024-05-21
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AU2020212111B2 (en) 2025-01-23
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