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WO2025098425A1 - 趋化因子受体调节剂制备方法和用途 - Google Patents

趋化因子受体调节剂制备方法和用途 Download PDF

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Publication number
WO2025098425A1
WO2025098425A1 PCT/CN2024/130449 CN2024130449W WO2025098425A1 WO 2025098425 A1 WO2025098425 A1 WO 2025098425A1 CN 2024130449 W CN2024130449 W CN 2024130449W WO 2025098425 A1 WO2025098425 A1 WO 2025098425A1
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Prior art keywords
compound
pharmaceutically acceptable
tautomer
stereoisomer
acceptable salt
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English (en)
French (fr)
Inventor
刘凤涛
李强
周晓波
张小娟
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Zhejiang Huahai Pharmaceutical Co Ltd
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Zhejiang Huahai Pharmaceutical Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • the invention belongs to the field of medicine and relates to a compound for regulating the activity of a chemokine receptor, a preparation method and use thereof.
  • Chemokines are a family of cytokines that guide leukocytes to move as chemical attractants. They are secreted by a variety of cells and can be functionally divided into two categories: hemostatic chemokines and inflammatory chemokines. Hemostatic chemokines are produced in certain tissues and control cells of the immune system during the immune surveillance process, such as directing lymphocytes to lymph nodes so that they can screen for pathogen invasion.
  • Inflammatory chemokines are released by cells in response to pathological events (e.g., proinflammatory stimulants, such as IL-1 or viruses). It mainly acts as a chemical attractant as a part of the inflammatory response, and is used to guide cells of the innate immune system and the adaptive immune system to the site of inflammation.
  • CC chemokine receptor type 4 (CCR4) plays a role in the progression of many inflammation-related and other diseases. Identifying compounds that regulate CCR4 function is a continuing challenge.
  • Cancer is a class of diseases in which cells exhibit dysregulated replication and growth. Recent cancer models implicate the immune system in the development and progression of cancer. While much progress has been made in understanding the biological basis of cancer, cancer remains a leading cause of death.
  • CCR4 was first identified by Power et al. (J. Biol. Chem. 270: 19495-19500) and is a G protein-coupled receptor that can bind to chemokines.
  • CC chemokines are chemotactic agents for the Th2 subset of peripheral blood T cells, dendritic cells, and natural killer cells.
  • Chemokines include CCL22 (also known as macrophage-derived chemokine, MDC).
  • CCL17 also known as thymus and activation-regulated chemokine, TARC, which is also produced by monocytes and dendritic cells.
  • CCR4 is involved in immune regulatory processes such as homing of cells to specific tissues, including homing of T lymphocytes to the skin and lungs (see, e.g., Campbell et al., 1999, Nature, 400:776-780; Gonzalo et al., 1999, J. Immunol, 163:403-411; Lloyd et al., 2000, J. Exp. Med, 191:265-273; Kawasaki et al., 2000, J. Immunol. 166:2055-2062). Regulators of CCR4 activity have been described, e.g., in WO2013/082490.
  • CTLA-4 Cytotoxic T lymphocyte antigen-4
  • CTLA-4 is considered a key regulator of adaptive immune responses, particularly in maintaining burst T cell responses and components used in burst T cell responses. Therefore, CTLA-4, as an immune checkpoint inhibitor, is considered a possible therapeutic target for the treatment of cancer and inflammation.
  • CTLA-4 modulators have been described, for example, in WO2018/035710.
  • PD-1 Programmed death receptor-1
  • CTLA4 PD-L1 and PD-L2. Similar to CTLA4, PD-1 is also a major regulator of the immune system and is also considered an immune checkpoint inhibitor.
  • PD-1/PD-L1 modulators have been described, for example, in WO2018/005374.
  • the first aspect of the present invention relates to a compound represented by formula (I), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof,
  • Ring A is a 4-8 membered aromatic heterocycle
  • X1 or X2 are each independently N or CH;
  • X3 is N or CR a ;
  • Y is -O- or -CH 2 -;
  • Ra is -H, -C1-3 alkyl, -halogen or -CN, wherein the -C1-3 alkyl is optionally substituted with one or more halogens within the range allowed by the valence;
  • L 1 is a bond, -C 1-3 alkylene-, -C 2-4 alkenylene- or -C 2-4 alkynylene-;
  • R1 is -C1-3alkylene -OH
  • R2 is -H, -CN, -C1-5 alkyl, 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl, the -C1-5 alkyl is optionally substituted with one or more halogens within the range allowed by the valence, the 3-8 membered cycloalkyl or 3-8 membered heterocycloalkyl is optionally substituted with one or more -C1-5 alkyl within the range allowed by the valence or -halogen;
  • R 4 are each independently halogen
  • n 0, 1, 2 or 3.
  • ring A is a 5-6 membered aromatic heterocycle; preferably, the heteroatom of the 5-6 membered aromatic heterocycle is N, O or S.
  • Ring A is wherein E 1 and E 2 are each independently CH, N, O or S, E 3 is a bond, CH or N, and E 1 , E 2 and E 3 are not CH at the same time.
  • Ring A is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • E1 is S, N, O
  • E2 is CH, N, O or S
  • E3 is a bond.
  • E1 is O, E2 is N, and E3 is a bond; or E1 is N, E2 is O, and E3 is a bond; or E1 is S, E2 is N, and E3 is a bond; or E1 is N, E2 is S, and E3 is a bond; or E1 is S, E2 is CH, and E3 is a bond.
  • E 1 , E 2 , and E 3 are each independently CH or N, and E 1 , E 2 , and E 3 are not CH at the same time.
  • E1 is CH, E2 is N, and E3 is CH; or E1 is CH, E2 is CH, and E3 is N; or E1 is N, E2 is N, and E3 is CH; E1 is CH, E2 is N, and E3 is N; or E1 is N, E2 is N, and E3 is CH; E1 is N, E2 is N, and E3 is CH; E1 is N, E2 is N, and E3 is N.
  • Ring A is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • Ring A is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • n 0, 1 or 2, preferably 2.
  • X1 or X2 are not N at the same time; preferably, X1 is CH, X2 is N, and X3 is N; or X1 is CH, X2 is N, and X3 is CR a ; or X1 is N, X2 is CH, and X3 is N; or X1 is CH, X2 is CH, and X3 is N; or X1 is CH, X2 is CH, and X3 is N; or X1 is CH, X2 is CH, and X3 is N; or X1 is CH, X2 is CH, and X3 is CR a .
  • Ra is -H or -CN.
  • R1 is -CH2CH2OH .
  • R2 is -H, -CN, -C1-5 alkyl, 3-5 membered cycloalkyl or 3-5 membered heterocycloalkyl
  • the -C1-5 alkyl is optionally substituted with one or more halogens within the range allowed by the valence
  • the 3-5 membered cycloalkyl or 3-5 membered heterocycloalkyl is optionally substituted with one or more -C1-3 alkyl or -halogens within the range allowed by the valence
  • the -C1-5 alkyl is methyl or tert-butyl
  • the 3-5 membered cycloalkyl is cyclopropyl
  • the 3-5 membered heterocycloalkyl is oxetanyl or azetidinyl.
  • R2 is -H, -CN, methyl, tert-butyl, cyclopropyl, oxetanyl or azetidinyl
  • the methyl or tert-butyl group is optionally substituted with one or more -F groups within the range allowed by the valence
  • the cyclopropyl, oxetanyl or azetidinyl group is optionally substituted with one or more methyl or -F groups within the range allowed by the valence.
  • R 2 is -H, -CN, -CH 3 , -CHF 2 , -CF 3 , -C(CH 3 ) 3 , cyclopropyl,
  • R 3 is -C 1-3 alkyl, preferably methyl.
  • R 4 is each independently -F, -Cl or -Br, preferably -Cl.
  • the compound represented by formula (I), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof has the following structure:
  • Y is -CH2-
  • X1 is CH
  • X2 is N
  • X3 is N.
  • the compound represented by formula (I), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof has the following structure:
  • R 3 is -H or -C 1-3 alkyl
  • R 5 is a 5-6 membered heteroaryl group optionally substituted by one or more substituents; the substituents are each independently selected from hydrogen, halogen, -CN, hydroxyl, -C 1-3 alkyl or -C 1-3 haloalkyl, and the 5-6 membered heteroaryl group contains 1, 2 or 3 heteroatoms.
  • the compound represented by formula (I), formula (II) or formula (III), its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, the compound represented by formula (I), formula (II) or formula (III) is selected from the compounds represented by the following structural formulas:
  • the second aspect of the present invention provides a method for preparing the compound described in the first aspect of the present invention, its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, comprising the steps shown in any one of the following schemes 1 to 3:
  • each group is as described above.
  • the third aspect of the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising the compound described in the first aspect of the present invention, its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, and optionally one or more pharmaceutically acceptable carriers and/or excipients.
  • the fourth aspect of the present invention provides use of the compound described in the first aspect of the present invention, its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, or the pharmaceutical composition described in the third aspect of the present invention in the preparation of a medicament for preventing and/or treating CCR4-mediated diseases.
  • the CCR4-mediated disease comprises an inflammatory disease, an autoimmune disease or cancer.
  • the inflammatory disease and autoimmune disease are dermatitis, allergic asthma, inflammatory bowel disease or lupus erythematosus
  • the cancer is liver cancer, pancreatic cancer, colon cancer, lung cancer, brain cancer or gastric cancer.
  • the fifth aspect of the present invention provides a method for treating CCR4-mediated diseases, comprising administering to a patient or individual in need of such treatment a therapeutically effective amount of any of the compounds described in the first aspect of the present invention, its stereoisomers, tautomers or pharmaceutically acceptable salts thereof, or the pharmaceutical composition described in the third aspect of the present invention.
  • optionally substituted methyl refers to "unsubstituted methyl” or “substituted methyl”.
  • -C 1-n alkyl refers to a non-cyclic, saturated, branched or straight-chain aliphatic hydrocarbon group having 1 to n carbon atoms, preferably an alkyl group having 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms (i.e., -C 1-12 alkyl), more preferably an alkyl group having 1 to 6 carbon atoms (i.e., -C 1-6 alkyl), and further preferably an alkyl group having 1 to 3 carbon atoms.
  • -C 1-3 alkyl refers to methyl, ethyl, n-propyl and isopropyl.
  • -C1 -n alkylene- refers to a residue resulting from the removal of a hydrogen atom from a -C1 -n alkyl group, which is a straight or branched chain group containing 1 to n carbon atoms, preferably containing 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms (i.e., -C1-12 alkylene-), and more preferably an alkylene group containing 1 to 6 carbon atoms ( i.e., -C1-6 alkylene-).
  • -C1-3 alkylene- includes -CH2- , -CH2 - CH2- , -CH( CH3 )-, -CH2 -CH2 - CH2- , -C( CH3 ) 2- , -CH( CH2CH3 )-, -CH( CH3 ) -CH2- , -CH2 -CH( CH3 )-, and the like.
  • An alkylene group may be substituted or unsubstituted, and when substituted, it may be substituted at any available point of attachment.
  • -C 2-n alkenyl refers to a group containing at least one carbon-carbon double bond in a -C 2-n alkyl group.
  • the alkenyl group contains 2 to 12 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12) carbon atoms (i.e., C 2-12 alkenyl), and more preferably contains 2 to 6 carbon atoms (i.e., C 2-6 alkenyl).
  • the alkenyl group may be substituted or unsubstituted, and when substituted, the substituent may optionally exist in cis or trans or E or Z form, for example:
  • -C2 -n alkenylene- refers to a residue derived from a -C2 -n alkenyl group by removing a hydrogen atom.
  • -C 2-n alkynylene- refers to a residue derived from a -C 2-n alkynyl group by removing a hydrogen atom.
  • -C 2-3 alkynylene- includes -C ⁇ C-, -C ⁇ C-CH 2 -, and -CH 2 -C ⁇ C-.
  • aromatic heterocycle refers to heterocyclic compounds with aromatic characteristics, including single heterocyclic aromatic groups and fused heterocyclic aromatic groups.
  • Typical 5-6 membered heteroaryl groups include, but are not limited to, 2- or 3-thienyl; 2- or 3-furyl; 2- or 3-pyrrolyl; 2-, 4- or 5-imidazolyl; 3-, 4- or 5-pyrazolyl; 2-, 4- or 5-thiazolyl; 3-, 4- or 5-isothiazolyl; 2-, 4- or 5-oxazolyl; 3-, 4- or 5-isoxazolyl; 3- or 5-1,2,4-triazolyl; 4- or 5-1,2,3-triazolyl; tetrazolyl; 2-, 3- or 4-pyridyl; 3- or 4-pyridazinyl; 3-, 4- or 5-pyrazinyl; 2-pyrazinyl; 2-, 4- or 5-pyrimidinyl.
  • cycloalkane refers to a saturated monocyclic or polycyclic ring
  • cycloalkyl refers to a saturated monocyclic or polycyclic cyclic alkyl substituent, the cycloalkyl ring containing 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 8 (e.g., 3, 4, 5, 6, 7 and 8) carbon atoms, more preferably 3 to 6 carbon atoms.
  • monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, and cyclopentyl.
  • heterocycloalkane refers to a cycloalkane comprising at least one ring heteroatom (e.g., a nitrogen atom, an oxygen atom, or a sulfur atom).
  • heterocycloalkyl refers to a cycloalkyl comprising at least one ring heteroatom (e.g., a nitrogen atom, an oxygen atom, or a sulfur atom).
  • the heterocyclyl is a 4-12 membered heterocyclyl.
  • the heterocyclyl is a 4-6 membered heterocyclyl, such as tetrahydrofuranyl, oxetane, or azetidine.
  • halo or halogen, by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.
  • isotopic derivative refers to a derivative in which one or more atoms in a compound are replaced by an isotope thereof (an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature).
  • isotopes include, but are not limited to, isotopes of hydrogen (e.g., 2 H, 3 H), isotopes of carbon (e.g., 11 C, 13 C, and 14 C), isotopes of fluorine (e.g., 18 F), isotopes of nitrogen (e.g., 13 N and 15 N), and isotopes of oxygen (e.g., 15 O, 17 O, and 18 O).
  • salts and “pharmaceutically acceptable salts” have the same meaning and refer to salts of the active compounds that are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein.
  • base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent.
  • pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts or similar salts.
  • acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent.
  • Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, and salts derived from relatively nontoxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, oxalic acid, methanesulfonic acid, etc.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid
  • salts of amino acids such as arginine salts, and salts of organic acids such as glucuronic acid or galacturonic acid, etc.
  • salts of amino acids such as arginine salts
  • organic acids such as glucuronic acid or galacturonic acid, etc.
  • Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.
  • “Pharmaceutically acceptable excipients” and “pharmaceutically acceptable carriers” refer to substances that facilitate administration of a compound to an individual and absorption by the individual and that can be included in the composition of the present invention without causing significant adverse toxic effects to the patient.
  • Non-limiting examples of pharmaceutically acceptable excipients include: Examples include water, NaCl, physiological saline solution, lactated Ringer's, ordinary sucrose, ordinary glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavoring agents, salt solutions (such as Ringer's), alcohols, oils, gelatin, carbohydrates (such as lactose, amylose or starch), fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone and pigments, etc.
  • Such preparations can be sterilized and, if necessary, can be mixed with auxiliary agents that do not adversely react with the compounds of the invention, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for affecting osmotic pressure, buffers, coloring and/or aromatic substances, etc.
  • auxiliary agents that do not adversely react with the compounds of the invention, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for affecting osmotic pressure, buffers, coloring and/or aromatic substances, etc.
  • treatment refers to any marker of successful treatment or improvement of an injury, disease, pathology, or condition, including any objective or subjective parameter, such as alleviation of symptoms; relief; reduction or making an injury, pathology, or condition more tolerable to the patient; slowing the rate of degeneration or decline; making the degenerative endpoint less debilitating; improving the patient's physical or mental health.
  • Treatment or improvement of symptoms can be based on objective or subjective parameters, including the results of a physical examination, neuropsychiatric examination, and/or psychiatric evaluation.
  • treatment and variations thereof may include prevention of an injury, pathology, condition, or disease. In embodiments, treatment is prevention. In embodiments, treatment does not include prevention.
  • Treatment includes preventive treatment.
  • the treatment method includes administering to an individual a therapeutically effective amount of a compound described herein.
  • the administration step may include a single administration or may include a series of administrations.
  • the length of the treatment period depends on a variety of factors, such as the severity of the condition, the age of the patient, the concentration of the compound, the activity of the composition used for treatment, and combinations thereof. It should also be understood that the effective amount of the agent used for treatment or prevention may increase or decrease over the course of a particular treatment or prevention regimen. Changes in dosage can be obtained and become apparent through standard diagnostic analyses known in the art. In some instances, long-term administration may be required.
  • the composition is administered to an individual in an amount and duration sufficient to treat the patient.
  • prevention refers to reducing the occurrence of disease symptoms in patients. As mentioned above, prevention can be complete (no detectable symptoms) or partial prevention, so that the observed symptoms are less than the symptoms that may occur when not treated. In embodiments, prevention refers to slowing down the progress of a disease, illness or condition, or inhibiting its progression to a harmful or other undesirable state.
  • "Patient” or “individual in need” refers to a living organism suffering from or susceptible to a disease or condition that can be treated by administering a pharmaceutical composition provided herein. Non-limiting examples include people, other mammals, cattle, rats, mice, dogs, monkeys, goats, sheep, cows, deer and other non-mammals. In some embodiments, the patient is a person.
  • an “effective amount” is an amount sufficient to achieve the specified purpose of the compound (e.g., to achieve the effect of its administration, treat a disease, reduce enzyme activity, increase enzyme activity, reduce signaling pathways, or alleviate one or more symptoms of a disease or condition) relative to the absence of the compound.
  • An “effective amount” example is an amount sufficient to promote the treatment, prevention, or alleviation of one or more symptoms of a disease, which may also be referred to as a "therapeutically effective amount.”
  • "Relief" of one or more symptoms means reducing the severity or frequency of the symptoms, or eliminating the symptoms.
  • a “prophylactic effective amount" of a drug is an amount that has an expected preventive effect when the drug is administered to an individual, such as preventing or delaying the onset (or recurrence) of damage, disease, pathology, or condition, or reducing the likelihood of onset (or recurrence) of damage, disease, pathology, or condition or its symptoms.
  • a complete preventive effect does not necessarily occur by administering one dose, and may only occur after a series of doses are administered. Therefore, a prophylactic effective amount may be administered during one or more administrations.
  • an “activity reduction amount” refers to the amount of antagonist required to reduce enzyme activity relative to the absence of an antagonist.
  • a “function-disrupting amount” refers to the amount of antagonist required to disrupt the function of an enzyme or protein relative to the absence of the antagonist. The exact amount will depend on the purpose of the treatment and will be determined by those skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (Volumes 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins). The therapeutically effective amount can be determined by measuring relevant physiological effects and can be adjusted in conjunction with individual dosing regimens and diagnostic analyses of the condition.
  • CCR4 inhibitor is a compound (eg, a compound described herein) that decreases CCR4 activity compared to a control (eg, a compound lacking the compound or known to be inactive).
  • administering means oral administration, suppository administration, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intracranial, intranasal or subcutaneous administration, or implanting a sustained release device such as a micro-osmotic pump into a subject.
  • Administration by any route, including parenteral and intramuscular administration, is contemplated.
  • Membranes e.g., oral, sublingual, palatal, gingival, nasal, vaginal, rectal or transdermal.
  • Parenteral administration includes, but is not limited to, intravenous, intramuscular, intraarteriolar, intradermal, subcutaneous, intraperitoneal, intraventricular and intracranial.
  • Combined administration refers to the administration of the compositions described herein at the same time, just before or just after the administration of one or more additional therapies (e.g., anticancer agents, chemotherapy or treatment of neurodegenerative diseases).
  • additional therapies e.g., anticancer agents, chemotherapy or treatment of neurodegenerative diseases.
  • the compounds of the present invention can be administered alone or can be administered to the patient in combination.
  • the preparation can also be combined with other active substances (e.g., to reduce metabolic degradation).
  • compositions of the present invention can be delivered transdermally by a local route and can be formulated into smear sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, coatings, powders and aerosols.
  • Oral formulations include tablets, pills, powders, dragees, capsules, liquids, lozenges, cachets, gels, syrups, slurries, suspensions, and the like suitable for ingestion by a patient.
  • Solid form formulations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules.
  • Liquid form formulations include solutions, suspensions, and emulsions, such as aqueous solutions or water/propylene glycol solutions.
  • the compositions of the present invention may additionally contain components that provide sustained release and/or comfort.
  • Such components include high molecular weight anionic mucomimetic polymers, gelling polysaccharides, and finely divided drug carrier matrices, as discussed in more detail in U.S. Patent Nos. 4,911,920, 5,403,841, 5,212,162, and 4,861,760. The entire contents of these patents are incorporated herein by reference in their entirety for all purposes.
  • the compositions of the present invention may also be delivered as microspheres for slow release in vivo.
  • the microspheres can be administered by intradermal injection of drug-containing microspheres that are slowly released subcutaneously (see Rao, J. Biomater Sci. Polym. 7th edition, 623-645, 1995; can be administered as a biodegradable and injectable gel formulation (see, e.g., Gao Pharm. Res. 12: 857-863, 1995); or as microspheres for oral administration (see, e.g., Eyles, J. Pharm. Pharmacol. 49: 669-674, 1997).
  • the formulation of the composition of the present invention can be delivered by using liposomes that fuse with cell membranes or are internalized by cells, that is, by using receptor ligands linked to liposomes that bind to surface membrane protein receptors of cells, thereby causing endocytosis.
  • liposomes especially carrying In the case of receptor ligands specifically targeting target cells or preferentially involving specific organs
  • the composition of the present invention can be delivered to the target cells in vivo.
  • the composition of the present invention can also be delivered as nanoparticles.
  • the first step is the synthesis of tert-butyl (R)-3-(1-(2-hydroxyethyl)piperidin-3-yl)azetidine-1-carboxylate 1c
  • Step 8 Synthesis of 2-((R)-3-(1-(1-(R)-1-(4,6-dichloropyridin-3-yl)ethyl)-3-((trimethylsilyl)ethynyl)-1H-pyrazolo[3,4-b]pyrazin-6-yl)azetidin-3-yl)piperidin-1-yl)ethane-1-ol 11
  • Example 8 2-((3R)-3-(1-(1-(2-(2,5-dichloro-1 ⁇ 3 ,3 ⁇ 2 -oxazol-4-yl)ethyl)-3-ethynyl-1H-pyrazolo[3,4-b]pyrazin-6-yl)azetidin-3-yl)piperidin-1-yl)ethan-1-ol (8)
  • Example 18 2-((3R)-3-(1-(1-((R)-1-(4,6-dichloropyridin-3-yl)ethyl)-3-((2,2-difluorocyclopropyl)ethynyl)-1H-pyrazolo[3,4-b]pyrazin-6-yl)azetidin-3-yl)piperidin-1-yl)ethan-1-ol (18)
  • Example 21 2-[(3R)-3-(1- ⁇ 1-[(1R)-1-(4,6-dichloropyridin-3-yl)ethyl]-3-(3,3,3-trifluoroprop-1-ynyl)pyrazolo[4,3-b]pyrazin-6-yl ⁇ azetidin-3-yl)hexahydropyridin-1-yl]ethan-1-ol (21)
  • Example 22 2-[(3R)-3- ⁇ 1-[3-(cyclopropylethynyl)-1-[(1R)-1-(4,6-dichloropyridin-3-yl)ethyl]pyrazolo[4,3-b]pyrazin-6-yl]azetidin-3-yl ⁇ hexahydropyridin-1-yl]ethan-1-ol (22)
  • Step 4 2-((R)-3-(1-(1-(R)-1-(3,5-dichloropyridin-2-yl)ethyl)-3-vinyl-1H-pyrazolo[4,3-b]pyridin-6-yl)azetidin-3-yl)piperidin-1-yl)ethane-1-ol (23)
  • Step 2 1-(((R)-1-(3,5-dichloropyridin-2-yl)ethyl)-6-(3-(((R)-1-(2-hydroxyethyl)piperidin-3-yl)azetidin-1-yl)-1H-pyrazolo[4,3-b]pyridine-3-carbonitrile (24)
  • Step 6 Synthesis of 26 g of 3-chloro-5-[(1R)-1-(4,6-dichloropyridin-3-yl)ethyl]-7-(trifluoromethyl)pyrrolo[3,2-b]pyrazine-6-carbonitrile
  • Example 27 2-[(3R)-3- ⁇ 1-[3-(cyclopropylethynyl)-1-[1-(2,4-dichloropyrimidin-5-yl)ethyl]pyrazolo[4,3-b]pyrazin-6-yl]azetidin-3-yl ⁇ hexahydropyridin-1-yl]ethan-1-ol (27)
  • Example 28 2-[(3R)-3-(1- ⁇ 3-[(1E)-2-cyclopropylvinyl]-1-[(1R)-1-(4,6-dichloropyridin-3-yl)ethyl]pyrazolo[4,3-b]pyrazin-6-yl ⁇ azetidin-3-yl)piperidin-1-yl]ethan-1-ol (28)
  • Example 32 2-[2-(1- ⁇ 1-[1-(4,6-dichloropyridin-3-yl)ethyl]-3-(trifluoromethyl)pyrazolo[4,3-b]pyrazin-6-yl ⁇ azetidin-3-yl)-1,4-oxazin-4-yl]ethan-1-ol (32)
  • Example 35 2-[(3R)-3- ⁇ 1-[3-(cyclopropylethynyl)-1-[1-(4,6-dichloropyridin-3-yl)ethyl]pyrazolo[4,3-c][1,2]diazin-6-yl]azetidin-3-yl ⁇ hexahydropyridin-1-yl]ethan-1-ol (35)
  • the first step is the synthesis of methyl 6-chloro-4-diazine-1,2-diazine-3-carboxylate 35b
  • Step 6 Synthesis of 35 g of 2-[(3R)-3- ⁇ 1-[3-(cyclopropylethynyl)-1-(5,5-dimethyl-2-oxo-5-silanyl-1-yl)pyrazolo[4,3-][1,2]diazine-6-yl]azetidin-3-yl ⁇ hexahydropyridin-1-yl]ethane-1-ol
  • Step 8 Synthesis of 2-[(3R)-3- ⁇ 1-[3-(cyclopropylethynyl)-1-[1-(4,6-dichloropyridin-3-yl)ethyl]pyrazolo[4,3-c][1,2]diazin-6-yl]azetidin-3-yl ⁇ hexahydropyridin-1-yl]ethane-1-ol 35
  • the compounds of the present invention have significant inhibitory activity on CCR4 receptor.
  • mice Female immunodeficient mice aged 8 weeks, with an average weight of 20 g.
  • the experiment began after the mice adapted for 3 days.
  • the experiment was divided into four groups, model group, FLX475 group and Example 1 compound group, with 8 mice in each group.
  • Humanized hepatoma cell Hepg2 was cultured before the experiment. After culturing to a certain number, 1*10 7 cells were injected subcutaneously to each mouse. After the tumor grew to a certain size, the model group, FLX475 group and Example compound 35 group were gavaged and injected at 10 mL/kg once a day; the model group was gavaged/injected with an equal amount of solvent for 28 consecutive days.
  • Table 3 The specific design is shown in Table 3.
  • mice Female immunodeficient mice, 8 weeks old. All mice were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd., license number SCXK (Shanghai) 2017-0011, animal certificate number: 20170011003865.
  • mice The tumor size and body weight of mice were observed daily, and the tumor size was measured three times a week.
  • mice in the model control group continued to increase with the number of days, while the tumor volume of Example Compound 35 began to decrease significantly after the 14th day.
  • HEK-293 cells stably expressing hERG potassium channel were cultured in DMEM medium containing 10% fetal bovine serum and 0.8 mg/mL G418 at 37°C and 5% carbon dioxide concentration.
  • Cell passaging Remove the old culture medium and wash once with PBS, then add 1 mL of TrypLE TM Express solution and incubate at 37°C for about 0.5 min. When the cells detach from the bottom of the dish, add about 5 mL of complete culture medium preheated at 37°C. Gently blow the cell suspension with a pipette to separate the aggregated cells. Transfer the cell suspension to a sterile centrifuge tube and collect the cells by centrifugation at 1000 rpm for 5 min. For expansion or maintenance culture, inoculate the cells in a 6 cm cell culture dish, with 2.5 ⁇ 105 cells inoculated in each cell culture dish (final volume: 5 mL).
  • the cell density must not exceed 80%.
  • the drug administration begins. After each drug concentration acts for 5 minutes (or the current stabilizes), the next concentration is detected. One or more concentrations are detected for each test compound.
  • the coverslip with cells is placed in the recording bath in an inverted microscope.
  • the test compound working solution and the external solution without the compound are administered by gravity perfusion from low concentration to high concentration through the recording bath to act on the cells.
  • a peristaltic pump is used for liquid replacement during recording.
  • the current detected by each cell in the external solution without the compound serves as its own control group. At least two cells are used for each concentration and the test is repeated twice independently. All electrophysiological experiments are performed at room temperature.
  • the tail current (Peak tail currentcompound) after each drug concentration and the tail current (Peak tail currentControl) of the blank solvent treatment group were normalized (Peak tail currentcompound/Peak tail currentControl), and then the inhibition rate corresponding to each drug concentration was calculated (1-(Peak tail currentcompound/Peak tail currentControl).
  • the mean (Mean), standard deviation (SD) and standard error (SE) of the inhibition rate were calculated, and the data were expressed as Mean ⁇ SE and saved in Excel.
  • Y 1/(1+10 ⁇ ((LogIC50-X)*HillSlope)
  • the IC50 value of each compound was calculated using the above equation, and the concentration effect curve was nonlinearly fitted, where IC50 was the half-inhibitory concentration.
  • the calculation of IC50 and curve fitting were completed using GraphPad Prism software.
  • the cardiac safety indicator hERG toxicity of the compound of the present invention is improved.

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Abstract

本发明提供了式(Ⅰ)所示的化合物、其立体异构体、互变异构体或其可药用盐，其制备方法、药物组合物及在制备用于预防和/或治疗CCR4介导的疾病的药物中的用途，属于医药领域。

Description

趋化因子受体调节剂制备方法和用途 技术领域
本发明属于医药领域，涉及用于调节趋化因子受体活性的化合物、其制备方法及用途。
背景技术
宿主防御系统的成功运作是若干过程共同进行消除外来病原体的结果。需要协调的固有免疫应答和获得性免疫应答，并且许多分泌因子和细胞相关因子已经被确认为协调和调节这两个宿主防御武器的重要介质。趋化因子是作为化学引诱物引导白细胞移动的细胞因子家族。它们是由多种细胞分泌，并且在功能上可以划分为两类：止血性趋化因子和炎性趋化因子。止血性趋化因子在某些组织中产生，并在免疫监视过程期间控制免疫系统的细胞，如将淋巴细胞导向至淋巴结使其能筛查病原体的入侵。炎性趋化因子是由细胞响应病理事件(例如，促炎性刺激物，如IL-1或病毒)而释放。其主要作为炎性反应的一部分的化学引诱物起作用，并用于引导固有免疫系统和适应性免疫系统的细胞至炎症部位。CC趋化因子受体4型(CCR4)在许多炎症相关和其它病症的进展中起作用。鉴定调节CCR4功能的化合物是一项持续的挑战。
癌症是细胞表现出复制和生长失调的一类疾病。最近的癌症模型显示免疫系统与癌症的发展和进展相关。虽然在了解癌症的生物学基础方面，研究已经取得了很大的进展，但癌症仍然是死亡的主要原因。
CCR4最早由Power等人鉴定(J.Biol.Chem.270:19495-19500)，是一种G蛋白偶联受体，可与趋化因子结合。CC趋化因子是外周血T细胞、树突状细胞和自然杀伤细胞的Th2子集的趋化剂。趋化因子包括CCL22(也称为巨噬细胞源趋化因子，MDC)。和CCL17(也称为胸腺和活化调节趋化因子，TARC)，其也由单核细胞和树突状细胞产生。
CCR4参与诸如细胞向特定组织的归巢，包括T淋巴细胞向皮肤和肺部的归巢等免疫调节过程(参见，例如：Campbell等人，1999，Nature，400:776-780；Gonzalo等人，1999，J.Immunol,163:403-411；Lloyd等人，2000,J.Exp.Med,191:265-273；Kawasaki等人，2000，J.Immunol.166:2055-2062)。CCR4活性的调节剂已在例如WO2013/082490中描述。
细胞毒性T淋巴细胞抗原-4(CTLA-4)被认为是适应性免疫反应的关键调节因子，特别是在维持突发性T细胞反应和突发性T细胞反应中所用组成成分中起着核心作用。因此，CTLA-4作为一种免疫检查点抑制剂，被认为是治疗癌症和炎症的可能治疗靶点。CLTA-4调节剂已在例如WO2018/035710中描述。
程序性死亡受体-1(PD-1)是一种跨膜受体蛋白，其通过与其两个天然配体PD-L1和PD-L2的相互作用，对T细胞的功能进行负调节。与CTLA4类似，PD-1也是免疫系统的主要调节因子，也被认为是免疫检查点抑制剂。PD-1/PD-L1调节剂已在例如WO2018/005374中描述。
鉴于细胞归巢和免疫检查点途径在癌症发展和进展中的作用，需要开发能够改善癌症治疗的联合疗法。
发明内容
本发明第一方面涉及式(Ⅰ)所示的化合物、其立体异构体、互变异构体或其可药用盐，
其中，环A为4-8元芳杂环；
X1或X2各自独立地为N或CH；
X3为N或CRa；
Y为-O-或-CH2-；
Ra为-H、-C1-3烷基、-卤素或-CN，所述-C1-3烷基任选地被一个或化合价所允许范围内的多个卤素所取代；
L1为键、-C1-3亚烷基-、-C2-4亚烯基-或-C2-4亚炔基-；
R1为-C1-3亚烷基-OH；
R2为-H、-CN、-C1-5烷基、3-8元环烷基或3-8元杂环烷基，所述-C1-5烷基任选地被一个或化合价所允许范围内的多个卤素取代，所述3-8元环烷基或3-8元杂环烷基任选地被一个或者化合价所允许范围内的多个-C1-5烷基或-卤素所取代；
R3为-C1-5烷基；
R4各自独立地为卤素；
n为0、1、2或3。
在一些实施方式中，环A为5-6元芳杂环；优选地，所述5-6元芳杂环的杂原子为N、O或S。
在一些实施方式中，环A为其中E1、E2各自独立地为CH、N、O或S，E3为键、CH或N，且E1、E2、E3不同时为CH。
在一些实施方式中，环A为
在一些实施方式中，E1为S、N、O，E2为CH、N、O或S，E3为键。
在一些实施方式中，E1为O，E2为N，E3为键；或E1为N，E2为O，E3为键；或E1为S，E2为N，E3为键；或E1为N，E2为S，E3为键；或E1为S，E2为CH，E3为键。
在一些实施方式中，E1、E2、E3各自独立地为CH或N，且E1、E2、E3不同时为CH。
在一些实施方式中，E1为CH，E2为N，E3为CH；或E1为CH，E2为CH，E3为N；或E1为N，E2为N，E3为CH；E1为CH，E2为N，E3为N；或E1为N，E2为N，E3为CH；E1为N，E2为N，E3为N。
在一些实施方式中，环A为
在一些实施方式中，环A为
在一些实施方式中，n为0、1或2，优选为2。
在一些实施方式中，X1或X2不同时为N；优选地，X1为CH，X2为N，X3为N；或X1为CH，X2为N，X3为CRa；或X1为N，X2为CH，X3为N；或X1为CH，X2为CH，X3为N；或X1为CH，X2为CH，X3为CRa。
在一些实施方式中，Ra为-H或-CN。
在一些实施方式中，L1为键、-C2-4亚烯基-或-C2-4亚炔基-，优选为键、-C≡C-或-CH＝CH-。
在一些实施方式中，R1为-CH2CH2OH。
在一些实施方式中，R2为-H、-CN、-C1-5烷基、3-5元环烷基或3-5元杂环烷基，所述-C1-5烷基任选地被一个或化合价所允许范围内的多个卤素取代，所述3-5元环烷基或3-5元杂环烷基任选地被一个或者化合价所允许范围内的多个-C1-3烷基或-卤素所取代；优选地，所述-C1-5烷基为甲基或叔丁基，所述3-5元环烷基为环丙基，所述3-5元杂环烷基为氧杂环丁基或氮杂环丁基。
在一些实施方式中，R2为-H、-CN、甲基、叔丁基、环丙基、氧杂环丁基或氮杂环丁基，所述甲基或叔丁基任选地被一个或化合价所允许范围内的多个-F取代，所述环丙基、氧杂环丁基或氮杂环丁基任选地被一个或者化合价所允许范围内的多个甲基或-F所取代。
在一些实施方式中，R2为-H、-CN、-CH3、-CHF2、-CF3、-C(CH3)3、环丙基、
在一些实施方式中，R3为-C1-3烷基，优选为甲基。
在一些实施方式中，R4各自独立地为-F、-Cl或-Br，优选为-Cl。
在一些实施方式中，式(Ⅰ)所示的化合物、其立体异构体、互变异构体或其可药用盐，具有以下结构：
其中，Y、L1、R2、E1～E3、X1～X3如式(Ⅰ)化合物中所述；
优选地，Y为-CH2-，X1为CH，X2为N，X3为N。
在一些实施方式中，式(Ⅰ)所示的化合物、其立体异构体、互变异构体或其可药用盐，具有以下结构：
其中，R3为-H或-C1-3烷基；
R5为任选被一个或多个取代基取代的5-6元杂芳基；所述取代基各自独立地选自氢、卤素、-CN、羟基、-C1-3烷基或-C1-3卤代烷基，所述5-6元杂芳基含1、2或3个杂原子。
在一些实施方式中，式(Ⅰ)、式(II)或式(Ⅲ)所示的化合物、其立体异构体、互变异构体或其可药用盐，所述式(Ⅰ)、式(II)或式(Ⅲ)所示的化合物选自以下结构式所示的化合物：

在一些实施方式中，式(Ⅰ)、式(II)或式(Ⅲ)所示的化合物、其立体异构体、互变异构体或其可药 用盐，所述式(Ⅰ)、式(II)或式(Ⅲ)所示的化合物选自以下结构式所示的化合物：

本发明第二方面提供本发明第一方面所述的化合物、其立体异构体、互变异构体或其可药用盐的制备方法，包含以下方案一～方案三中任一方案所示步骤：
方案一：
方案二：当X3为CRa时
方案三：当X3为N时
其中，各基团如前所述。
本发明第三方面提供一种药物组合物，包括本发明第一方面所述的化合物、其立体异构体、互变异构体或其可药用盐，以及任选一种或多种药学上可接受的载体和/或赋形剂。
本发明第四方面提供本发明第一方面所述的化合物、其立体异构体、互变异构体或其可药用盐，或者本发明第三方面所述的药物组合物在制备用于预防和/或治疗CCR4介导的疾病的药物中的用途。
在本发明的一些实施方案中，所述CCR4介导的疾病包括炎性疾病、自身免疫性疾病或癌症。
在本发明的一些实施方案中，所述炎性疾病和自身免疫疾病为皮炎、过敏性哮喘、炎症性肠病或红斑狼疮，所述癌症为肝癌、胰腺癌、结肠癌、肺癌、脑癌或胃癌。
本发明第五方面提供治疗CCR4介导的疾病的方法，包括给需要这种治疗的患者或有需要的个体施用治疗有效量的任一本发明第一方面所述的化合物、其立体异构体、互变异构体或其可药用盐，或者本发明第三方面所述的药物组合物。
具体实施方式
除非另有说明，否则采用本领域技术范围内的常规方法，如质谱、NMR、和药理学方法。除非提出具体定义，否则本文在分析化学、有机合成化学以及药物和药物化学的有关描述中采用的术语是本领域已知的。可在化学合成、化学分析、药物制备、制剂和递送，以及对患者的治疗中使用标准技术。通常可根据本说明书中引用和讨论的多个概要性和较具体的文献中的描述，按照本领域熟知的常规方法实施上述技术和方法。
除非有相反陈述，在说明书和权利要求书中使用的术语具有下述含义。
术语“任选地”包括“可选”和“不选”两种情况。例如，“任选取代的甲基”是指“未取代的甲基”或者“被取代的甲基”。术语“取代”是指所指定原子或基团上的任一或多个氢被指定基团的选择替代，条件为不超过所指定原子的正常价态。当取代基是氧代或酮基(即＝O)时，则所述原子上的2个氢被替代。
术语“-C1-n烷基”，(其中n为2至n的整数)表示具有1至n个碳原子的非环状、饱和、支链或直链脂肪族烃基团，优选含有1至12个(例如1、2、3、4、5、6、7、8、9、10、11和12个)碳原子的烷基(即-C1-12烷基)，更优选为含有1至6个碳原子的烷基(即-C1-6烷基)，进一步优选为含有1至3个碳原子的烷基。例如-C1-3烷基指甲基、乙基、正丙基和异丙基。
术语“-C1-n亚烷基-”是指从-C1-n烷基上除去一个氢原子产生的残基，其为包含1至n个碳原子的直链或支链基团，优选含有1至12个(例如1、2、3、4、5、6、7、8、9、10、11和12个)碳原子(即-C1-12亚烷基-)，更优选含有1至6个碳原子的亚烷基(即-C1-6亚烷基-)。例如，-C 1-3亚烷基-包括-CH2-、-CH2-CH2-、-CH(CH3)-、-CH2-CH2-CH2-、-C(CH3)2-、-CH(CH2CH3)-、-CH(CH3)-CH2-、-CH2-CH(CH3)-等。亚烷基可以是取代的或非取代的，当被取代时，其可以在任何可使用的连接点上被取代。
术语“-C2-n烯基”指-C2-n烷基中含有至少一个碳碳双键的基团。优选含有2至12个(例如2、3、4、5、6、7、8、9、10、11和12个)碳原子的烯基(即C2-12烯基)，更优选含有2至6个碳原子的烯基(即C2- 6烯基)。例如，术语-C2-3烯基包括-CH＝CH2、-CH＝CH-CH3、-CH2-CH＝CH2。烯基可以是取代的或非取代的，当被取代时，取代基可任选地以顺式或反式或者E或Z形式存在，例如：
术语“-C2-n亚烯基-”指从-C2-n烯基上除去一个氢原子产生的残基，例如，术语-C2-3亚烯基-包括-CH＝CH-、-CH＝CH-CH2-、-CH2-CH＝CH-。
术语“-C2-n-炔基”指-C2-n烷基中含有至少一个碳碳叁键的基团。互相结合。例如，术语-C2-3炔基包括-C≡CH、-C≡C-CH3、-CH2-C≡CH。
术语“-C2-n亚炔基-”指从-C2-n炔基上除去一个氢原子产生的残基，例如，-C2-3亚炔基-包括-C≡C-、-C≡C-CH2-、-CH2-C≡C-。
指存在键或不存在键，例如指单键或双键
术语“芳杂环”、“芳杂环基”、“杂芳环”或“杂芳环基”具有相同的含义，是指有芳香特征的杂环化合物，包括单杂环芳基和稠杂环芳基。典型的5-6元杂芳基包括但不限于，2-或3-噻吩基；2-或3-呋喃基；2-或3-吡咯基；2-、4-或5-咪唑基；3-、4-或5-吡唑基；2-、4-或5-噻唑基；3-、4-或5-异噻唑基；2-、4-或5-唑基；3-、4-或5-异唑基；3-或5-1，2，4-三唑基；4-或5-1，2，3-三唑基；四唑基；2-、3-或4-吡啶基；3-或4-哒嗪基；3-、4-或5-吡嗪基；2-吡嗪基；2-、4-或5-嘧啶基。
术语“环烷”指饱和的单环或多环，术语“环烷基”指饱和的单环或多环环状烷取代基，环烷基环包含3至20个碳原子，优选包含3至12个碳原子，更优选包含3至8个(例如3、4、5、6、7和8个)碳原子，更优选包含3至6个碳原子。单环环烷基的非限制性实例包括但不限于环丙基、环丁基、环戊基。
术语“杂环烷”是指包含至少一个环杂原子(例如，氮原子、氧原子或硫原子)的环烷。术语“杂环烷基”是指包含至少一个环杂原子(例如，氮原子、氧原子或硫原子)的环烷基。在本发明的一些实施例中，杂环基为4-12元杂环基。在本发明的一些实施例中，杂环基为4-6元杂环基，例如四氢呋喃基、氧杂环丁烷或氮杂环丁烷。
除另有规定，否则术语“卤代”或“卤素”，本身或作为另一取代基的一部分，意指氟、氯、溴或碘原子。
术语“同位素衍生物”指化合物中一个或多个原子被其同位素(具有相同原子序数但原子质量或质量数不同于在自然界中占优势的原子质量或质量数的原子)替代的衍生物。同位素的实例包括但不限于，氢的同位素(例如2H、3H)、碳的同位素(例如11C、13C及14C)、氟的同位素(例如18F)、氮的同位素(例如13N及15N)、氧的同位素(例如15O、17O及18O)。
术语“可药用盐”、“药学上可接受的盐”具有相同的含义，指包括根据本文描述的化合物上发现的特定取代基而定，用相对无毒的酸或碱制备的活性化合物的盐。当本发明化合物含有相对酸性的官能团时，可以通过使中性形式的此类化合物与足量的所需碱(纯的或在合适的惰性溶剂中)接触而获得碱加成盐。药学上可接受的碱加成盐的实例包括钠、钾、钙、铵、有机氨基或镁盐或类似的盐。当本发明化合物含有相对碱性的官能团时，可以通过使中性形式的此类化合物与足量的所需酸(纯的或在合适的惰性溶剂中)接触而获得酸加成盐。药学上可接受的酸加成盐的实例包括源自无机酸的那些盐，所述无机酸如盐酸、氢溴酸、硝酸、碳酸、一氢碳酸、磷酸、一氢磷酸、二氢磷酸、硫酸、一氢硫酸、氢碘酸或亚磷酸等，以及源自相对无毒的有机酸的盐，所述有机酸如乙酸、丙酸、异丁酸、马来酸、丙二酸、苯甲酸、琥珀酸、辛二酸、富马酸、乳酸、扁桃酸、邻苯二甲酸、苯磺酸、对甲苯磺酸、枸橼酸、酒石酸、草酸、甲磺酸等。还包括氨基酸的盐如精氨酸盐等，以及有机酸如葡糖醛酸或半乳糖醛酸等的盐(参见，例如Berge等人，“Pharmaceutical Salts”,Journal of Pharmaceutical Science,1977,66,1-19)。本发明的某些特定化合物含有碱性及酸性官能团两者，这允许化合物转变为碱或酸加成盐。
“药学上可接受的赋形剂”和“药学上可接受的载体”指助于向个体给药化合物并被个体吸收，并且可以包括在本发明的组合物中，不会对患者引起显著不良毒性作用的物质。药学上可接受的赋形剂的非限制性 实例包括水、NaCl、生理盐水溶液、乳酸林格氏液(lactated Ringer’s)、普通蔗糖、普通葡萄糖、粘合剂、填充剂、崩解剂、润滑剂、包衣、甜味剂、调味剂、盐溶液(如林格氏液)、醇、油、明胶、碳水化合物(如乳糖、直链淀粉或淀粉)、脂肪酸酯、羟甲基纤维素、聚乙烯吡咯烷酮和色素等。此类制剂可灭菌，并且若需要，可与不会与本发明化合物发生有害反应的助剂如润滑剂、防腐剂、稳定剂、润湿剂、乳化剂、用于影响渗透压的盐、缓冲剂、着色和/或芳香物质等混合。本领域技术人员会认识到药物赋形剂在本发明有用。
术语“治疗”是指成功治疗或改善损伤、疾病、病理或病状的任何标记，包括任何客观或主观参数，如症状的减轻；缓解；减少或者使得损伤、病理或病状更为患者所耐受；退化或衰退速率减缓；使得退化终点衰弱程度较小；改善患者的身体或精神健康。症状的治疗或改善可以基于客观或主观参数，包括身体检查、神经精神检查和/或精神病学评价的结果。术语“治疗”及其变形可包括预防损伤、病理、病状或疾病。在实施方案中，治疗是预防。在实施方案中，治疗不包括预防。
本文所用的“治疗”(且如本领域公知的)还广泛地包括任何可在个体的病状上获得有利或期望结果(包括临床结果)的方法。有利或期望临床结果包括但不限于减轻或改善一或多种症状或病状、减少疾病程度、稳定疾病状态(即不恶化)、预防疾病传播或扩散、延迟或减缓疾病进展、改善或缓和疾病状态、减少疾病复发及缓解，不论部分或完全及不论可检测或不可检测。换言之，本文所用“治疗”包括疾病的任何治愈、改善或预防。治疗可预防该疾病发生；抑制该疾病扩散；减轻该疾病的症状、完全或部分去除该疾病的根本原因、缩短疾病持续时间或这些事件的组合。
“治疗”包括预防性治疗。治疗方法包括向个体给药治疗有效量的本文所述的化合物。给药步骤可以包括单次给药或可以包括一系列给药。治疗期长度取决于多种因素，如病状严重性、患者年龄、化合物浓度、用于治疗的组合物的活性及其组合。还应理解，用于治疗或预防的剂的有效量可随特定治疗或预防方案的过程而增加或减少。通过本领域已知的标准诊断分析可以得到剂量的改变并且变得显而易见。在一些实例中，可能需要长期给药。例如，将组合物以足以治疗患者的量及持续时间向个体给药。
术语“预防”是指减少患者的疾病症状发生。如上所述，预防可以为完全(没有可检测的症状)或部分预防，使得观察到的症状比不治疗时可能发生的症状更少。在实施方案中，预防是指减缓疾病、病症或病状的进展，或者抑制其进展到有害或其它不期望的状态。“患者”或“有需要的个体”是指患有或易患可通过给药本文所提供的药物组合物来治疗的疾病或病状的活生物体。非限制性实例包括人、其它哺乳动物、牛、大鼠、小鼠、狗、猴子、山羊、绵羊、奶牛、鹿和其它非哺乳类动物。在一些实施方案中，患者是人。
“有效量”是相对于不存在化合物足以使化合物实现规定目的(例如达到其给药的效果，治疗疾病，降低酶活性，增加酶活性，减少信号通路或者减轻疾病或病状的一种或多种症状)的量。“有效量”实例是足以促进疾病的一种或多种症状的治疗、预防或减轻的量，也可称为“治疗有效量”。一种或多种症状的“减轻”(以及该短语的语法同等词)意指降低症状的严重程度或频率，或消除症状。药物的“预防有效量”是药物向个体给药时会具有预期的预防效果的量，所述预防效果例如预防或延迟损伤、疾病、病理或病状的发作(或复发)，或降低损伤、疾病、病理或病状或其症状发作(或复发)的可能性。完全的预防效果不一定通过给药一个剂量而出现，并且可能只有在给药一系列剂量之后出现。因此，预防有效量可以是在一次或多次给药的过程中给药的。如本文中所用，“活性减少量”指相对于不存在拮抗剂降低酶活性需要的拮抗剂的量。如本文中所用，“功能破坏量”指相对于不存在拮抗剂破坏酶或蛋白质的功能需要的拮抗剂的量。确切的量会取决于治疗目的，并且会由本领域技术人员使用已知技术确定(参见，例如，Lieberman,Pharmaceutical Dosage Forms(第1-3卷，1992)；Lloyd,The Art,Science and Technology of Pharmaceutical Compounding(1999)；Pickar,Dosage Calculations(1999)；及Remington:The Scienceand Practice of Pharmacy，第20版，2003,Gennaro,Ed.,Lippincott,Williams&Wilkins)。治疗有效量可以通过测量相关生理效应来确定，并且可以结合个体的病状的给药方案和诊断分析等进行调整。例如在给药后的特定时间测量CCR4抑制剂(或者，例如，其代谢物)的血清水平可指示是否已给药治疗有效量。“CCR4抑制剂”是指与对照(如不含该化合物或已知无活性的化合物)相比降低CCR4活性的化合物(例如本文所述的化合物)。
术语“给药”意指口服给药、栓剂给药、局部接触、静脉内、肠胃外、腹膜内、肌内、病灶内、鞘内、颅内、鼻内或皮下给药，或向个体植入缓释装置，如微量渗透泵。通过任何途径给药，包括肠胃外和经粘 膜(例如，口腔、舌下、腭、齿龈、鼻部、阴道、直肠或透皮)。肠胃外给药包括但不限于静脉内、肌内、小动脉内、皮内、皮下、腹膜内、心室内和颅内。其它递送模式包括但不限于使用脂质体制剂、静脉输注、透皮贴剂等。“联合给药”指在给药一种或多种附加疗法(例如，抗癌剂、化疗或治疗神经退行性疾病)的同时、就在之前或就在之后给药本文描述的组合物。本发明的化合物可以单独给药或可以向患者联合给药。需要时，也可将制剂与其它活性物质组合(例如减少代谢降解)。本发明的组合物可以通过局部途径透皮递送，可配制成涂抹棒、溶液剂、混悬剂、乳剂、凝胶剂、霜剂、软膏剂、糊剂、胶冻剂、涂布剂、散剂和气雾剂。口服制剂包括适合患者摄取的片剂、丸剂、散剂、糖衣丸、胶囊剂、液体、锭剂、扁囊剂、凝胶剂、糖浆剂、浆液、混悬剂等。固体形式制剂包括散剂、片剂、丸剂、胶囊剂、扁囊剂、栓剂和分散性颗粒剂。液体形式制剂包括溶液剂、混悬剂和乳剂，例如水溶液剂或水/丙二醇溶液剂。本发明的组合物可以另外包含提供持续释放和/或舒适性的组分。此类组分包括高分子量的阴离子类粘液状(mucomimetic)聚合物、胶凝多糖和细碎的药物载体基质，在美国专利No.4,911,920、5,403,841、5,212,162和4,861,760中有更详细的讨论。这些专利的全部内容整体援引加入本文，用于所有目的。本发明的组合物也可以作为微球体递送，以便在体内缓慢释放。例如，微球体可以通过皮内注射在皮下缓慢释放的含药微球体而给药(参见Rao,J.Biomater Sci.Polym.第7版，623-645,1995；可以作为可生物降解和可注射的凝胶制剂而给药(参见例如Gao Pharm.Res.12:857-863,1995)；或者作为用于口服给药的微球体而给药(参见例如Eyles,J.Pharm.Pharmacol.49:669-674,1997)。在另一个实施方案中，本发明组合物的制剂可以通过使用与细胞膜融合或被细胞内吞的脂质体来递送，即，通过采用与脂质体连接的受体配体来递送，这些受体配体结合到细胞的表面膜蛋白受体，从而引起细胞内吞作用。通过使用脂质体，特别是在脂质体表面携带了特别针对靶细胞或者说是优先涉及特定器官的受体配体的情况下，可以集中地将本发明的组合物递送到体内的靶细胞中。(参见例如，Al-Muhammed,J.Microencapsul.13:293-306,1996；Chonn,Curr.Opin.Biotechnol.6:698-708,1995；Ostro,Am.J.Hosp.Pharm.46:1576-1587,1989)。本发明的组合物也可以作为纳米粒递送。
通过以下实施例进一步举例描述本发明，这些方案是说明性的，并不以任何方式限制本发明。对本发明所作的本领域普通技术人员容易实现的任何改动或改变都将落入本发明的范围内。
本发明实施例中所用的原料或试剂除特别说明之外，均市售可得。
本发明中使用的缩写具有本领域常规含义，例如以下缩写的含义如下：

化合物的制备方法
实施例1：2-((R)-3-(1-(1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(1)
第一步(R)-3-(1-(2-羟乙基)哌啶-3-基)氮杂环丁烷-1-羧酸叔丁酯1c的合成
将化合物1a(4.0g，17mmol)溶于乙腈(40mL)中，加化合物1b(4.3g，25mmol)，碳酸铯(8.1g，25mmol)室温搅拌过夜，反应完毕，旋干，加入乙酸乙酯，水洗，水相乙酸乙酯萃取两次，合并有机相，水洗，饱和食盐水洗，无水硫酸钠干燥，旋干得到化合物1c(4.7g)。LCMS(ESI-MS)m/z:285.1[M+H]+.
第二步(R)-2-(3-(氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇三氟乙酸盐1d的合成
将化合物1c(4.7g，17mmol)溶于DCM(20mL)，加TFA(10mL)，室温下搅拌30分钟，TLC检测，碘缸显色，反应完毕，旋干，甲苯带干，得到化合物1d(8.2g)。LCMS(ESI-MS)m/z:185.1[M+H]+.
第三步4,6-二氯-N-甲氧基-N-甲基烟酰胺1f的合成
将化合物1e(10g，52mmol)溶于DMF(40mL)中，加甲氧基甲胺(4.8g，79mmoL)，DIPEA(14g，100mol)，分批加入HATU(30g,79mmol)室温搅拌1小时，LCMS检测反应完毕，乙酸乙酯萃取，有机相过Flash柱(石油醚/乙酸乙酯＝0-20％)得化合物1f(10g)。LCMS(ESI-MS)m/z:235.0[M+H]+.
第四步1-(4,6-二氯吡啶-3-基)乙烷-1-酮1g的合成
将化合物1f(10g,43mmol)溶于THF(5.0mL)，冰浴加入甲基溴化镁(29mL,86mmol)，搅拌反应1小时，基本反应完毕，饱和氯化铵的水溶液淬灭，乙酸乙酯萃取，有机相水洗，饱和食盐水洗过Flash柱(石油醚/乙酸乙酯＝0-10％)，得化合物1g(7.0g)。LCMS(ESI-MS)m/z:190.0[M+H]+.
第五步(S)-1-(4,6-二氯吡啶-3-基)乙烷-1-醇1h的合成
化合物1g(3.5g，19mmol)溶于THF(20mL)中-78℃下，半小时内缓慢滴加入(-)-IPC2Bci(13mL,22mmol)，缓慢升到室温搅拌3小时，LCMS检测，反应完毕，水淬灭，乙酸乙酯萃取，有机相水洗，饱 和食盐水洗，无水硫酸钠干燥过Flash柱得化合物1h(3.3g)。LCMS(ESI-MS)m/z:192.0[M+H]+.
第六步(R)-6-氯-1-(1-(4,6-二氯吡啶-3-基)乙基)-3-碘-1H-吡唑并[3,4-b]吡嗪1j的合成
化合物1i(4.0g，14mmol)溶于THF(20mL)中，冰浴下加入化合物1h(3.3g,17mmol)，PPh3(4.3g,21mmol)，氮气保护下加入DIAD(5.6g,21mmol)，室温搅拌过夜，LCMS检测，反应完毕，用水淬灭，乙酸乙酯萃取，有机相水洗，饱和食盐水洗，无水硫酸钠干燥，过Flash柱得化合物1j(3.0g)。LCMS(ESI-MS)m/z:454.0[M+H]+.
第七步2-((R)-3-(1-(1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-3-碘-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇1k的合成
化合物1j(2.5g，5.5mmol)溶于乙腈(30mL)中，冰浴下加入化合物1d(59％，3.9g，8.3mmol)，碳酸铯(9.0g,28mmol)，室温搅拌2小时，LCMS检测，反应完毕，水淬灭，乙酸乙酯萃取，有机相水洗，饱和食盐水洗，无水硫酸钠干燥过Flash柱得化合物1k(2.2g)LCMS(ESI-MS)m/z:602.2[M+H]+.
第八步2-((R)-3-(1-(1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-3-((三甲基甲硅烷基)乙炔基)-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇1l的合成
化合物1k(2.2g，3.7mmol)溶于THF(25mL)中，加入Pd(PPh3)Cl2(260mg,0.37mmol)，CuI(140mg,0.73mmol)，三乙胺(740mg,7.3mmol)，氮气保护下加入TMS炔烃(1.4g,15mmol)的THF(5.0mL)溶液，40℃加热反应15分钟，点板反应完毕，过滤旋干过Flash柱得化合物1l(1.6g)。LCMS(ESI-MS)m/z:572.3[M+H]+.
第九步2-((R)-3-(1-(1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇1的合成
化合物1l(1.1g，1.8mmol)溶于甲醇(10mL)中，加入K2CO3(770mg,5.6mmol)，室温搅拌十分钟，点板反应完毕，过滤，旋干，过Flash柱得到目标化600mg送SFC拆分，最终得到化合物1(430mg)。LCMS(ESI-MS)m/z:500.2[M+H+).
1H NMR(400MHz,DMSO-d6)δ8.57(s,1H),7.89(s,1H),7.81(s,1H),6.20(m,1H),4.53(s,1H),4.33(m,1H),4.16(m,2H),3.87(m,2H),3.48(m,2H),2.75(m,2H),2.56(m,2H),2.36(m,2H),1.89(m,1H),1.85(d,J＝8.0Hz,3H),1.64(m,4H),1.43(m,1H).
实施例2：2-((3R)-3-(1-(1-(2-(2,6-二氯吡啶-3-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(2)
化合物2的制备参照实施例1。LCMS(ESI-MS)m/z:500.30[M+H]+.
1H NMR(400MHz,DMSO-d6)δ7.98(dd,J＝8.2,3.1Hz,1H),7.88(s,1H),7.59(d,J＝8.1Hz,1H),6.17(q,J＝7.0Hz,1H),4.53(s,1H),4.16(d,J＝9.3Hz,2H),3.87(s,3H),3.47(t,J＝6.3Hz,2H),2.80–2.68(m,2H),2.54(s,1H),2.35(t,J＝6.3Hz,2H),1.92(t,J＝11.1Hz,1H),1.80(d,J＝7.0Hz,3H),1.66(td,J＝30.0,27.3,13.5Hz,4H),1.43(t,J＝11.8Hz,1H),0.82(d,J＝12.0Hz,1H).
实施例3：2-((3R)-3-(1-(1-(2-(2,4-二氯嘧啶-5-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(3)
化合物3的制备参照实施例1。LCMS(ESI-MS)m/z:501.40[M+H]+.
1H NMR(400MHz,DMSO-d6)δ9.03(d,J＝3.0Hz,1H),7.90(s,1H),6.12(q,J＝7.0Hz,1H),4.57(s,1H),4.17(s,2H),3.87(s,2H),3.54(t,J＝6.0Hz,2H),2.93(d,J＝10.8Hz,2H),2.61–2.51(m,3H),2.15(d,J＝11.8Hz,1H),1.85(d,J＝7.1Hz,5H),1.65(d,J＝15.1Hz,3H),1.50(d,J＝12.7Hz,1H),0.94–0.82(m,1H).
实施例4：2-((3R)-3-(1-(1-(4-(4,6-二氯哒嗪-3-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(4)
化合物4的制备参照实施例1。LCMS(ESI-MS)m/z:501[M+H]+.
实施例5：2-((3R)-3-(1-(1-(3-(3,5-二氯吡嗪-2-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(5)
化合物5的制备参照实施例1。LCMS(ESI-MS)m/z:501.40[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.87(s,1H),7.88(s,1H),6.25(q,J＝6.8Hz,1H),4.50(s,1H),4.35(s,1H),4.12(s,2H),3.82(s,2H),3.48(t,J＝6.3Hz,2H),2.75(d,J＝10.5Hz,2H),2.55(d,J＝7.0Hz,1H),2.36(t,J＝6.3Hz,2H),1.94(t,J＝11.2Hz,1H),1.81(d,J＝6.8Hz,3H),1.75–1.55(m,4H),1.42(d,J＝12.1Hz,1H),0.83(d,J＝11.3Hz,1H).
实施例6：2-((3R)-3-(1-(1-(3-(3,5-二氯-1,2,4-三嗪-6-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基]氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(6)
化合物6的制备参照实施例1。LCMS(ESI-MS)m/z:502[M+H]+.
实施例7：2-((3R)-3-(1-(1-(3-(3,5-二氯吡啶-2-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(7)
化合物7的制备参照实施例1。LCMS(ESI-MS)m/z:500.4[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.61(d,J＝2.2Hz,1H),8.23(d,J＝2.2Hz,1H),7.88(s,1H),6.27(q,J＝6.8Hz,1H),4.49(s,1H),4.15(s,2H),3.85(s,2H),3.65(s,1H),3.50(t,J＝6.2Hz,2H),2.79(d,J＝10.1Hz,2H),2.55(d,J＝7.7Hz,1H),2.41(t,J＝6.3Hz,2H),1.98(t,J＝10.9Hz,1H),1.81(d,J＝6.9Hz,3H),1.77–1.52(m,4H),1.46(t,J＝12.2Hz,1H),0.84(d,J＝11.8Hz,1H).
实施例8：2-((3R)-3-(1-(1-(2-(2,5-二氯-1λ3,3λ2-恶唑-4-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(8)
化合物8的制备参照实施例1。LCMS(ESI-MS)m/z:490[M+H]+.
实施例9：2-((3R)-3-(1-(1-(2-(2,4-二氯恶唑-5-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(9)
化合物9的制备参照实施例1。LCMS(ESI-MS)m/z:490[M+H]+.
实施例10：2-((3R)-3-(1-(1-(2-(2,4-二氯噻唑-5-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(10)
化合物10的制备参照实施例1。LCMS(ESI-MS)m/z:506[M+H]+.
实施例11：2-((3R)-3-(1-(1-(2-(2,5-二氯噻唑-4-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(11)
化合物11的制备参照实施例1。LCMS(ESI-MS)m/z:506[M+H]+.
实施例12：2-((3R)-3-(1-(1-(2-(2,5-二氯噻吩-3-基)乙基)-3-乙炔基-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(12)
化合物12的制备参照实施例1。LCMS(ESI-MS)m/z:505.3[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.01(s,1H),7.23(s,1H),6.04-6.01(m,1H),5.32(s,1H),4.72(s,1H),4.19(s2H),3.90-3.88(m,2H),3.57-3.55(m,2H),3.01(s,2H),2.65(s,2H),2.58-2.54(m,2H),1.89(s,1H),1.80-1.79(d,3H),1.74-1.71(m,2H),1.54-1.52(m,2H),0.94-0.91(m,1H).
实施例13：2-((R)-3-(1-(1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-3-(丙-1-炔-1-基)-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(13)
化合物13的制备参照实施例1。LCMS(ESI-MS)m/z:514[M+H]+.
实施例14：2-((R)-3-(1-(1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-3-(3,3-二甲基丁-1-炔-1-基)-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(14)
化合物13的制备参照实施例1。LCMS(ESI-MS)m/z:556.23[M+H]+.
实施例15：2-[(3R)-3-{1-[3-(环丙基乙炔基)-1-[(1R)-1-(4,6-二氯吡啶-3-基)乙基]吡唑并[4,3-b]吡嗪-6-基]氮杂环丁烷-3-基}六氢吡啶-1-基]乙烷-1-醇(15)
化合物15的制备参照实施例1。LCMS(ESI)m/z:540.40[M+H]+。
实施例16：2-((R)-3-(1-(1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-3-(氧杂环丁烷-3-基乙炔基)-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁-3-基)哌啶-1-基)乙烷-1-醇(16)
化合物16的制备参照实施例1。LCMS(ESI-MS)m/z:556.23[M+H]+.
实施例17：2-((R)-3-(1-(1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-3-((1-甲基氮杂环丁烷-3-基)乙炔基)-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷3-基)哌啶-1-基)乙烷-1-醇(17)
化合物17的制备参照实施例1。LCMS(ESI-MS)m/z:569.23[M+H]+.
实施例18：2-((3R)-3-(1-(1-((R)-1-(4,6-二氯吡啶-3-基)乙基)-3-((2,2-二氟环丙基)乙炔基)-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(18)
化合物18的制备参照实施例1。LCMS(ESI-MS)m/z:576.23[M+H]+.
实施例19：2-((R)-3-(1-(1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-3-(三氟甲基)-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(19)
第一步(R)-6-氯-1-(1-(4,6-二氯吡啶-3-基)乙基)-3-(三氟甲基)-1H-吡唑并[3,4-b]吡嗪19a的合成
将化合物1j(5.0g，11mmol)，氟磺酰基二氟乙酸甲酯(4.2g,22mmol)，六甲基磷酰三胺(7.9g,44mmol)，CuI(2.1g，11mmol)加入单口瓶，加入DMF(50mL)，置换氮气，100℃加热反应4h。反应液降至室温，倒入水中，析出固体，抽滤，固体二氯甲烷洗三次，合并有机相，浓缩过flash柱子(流动相PE:EA＝5:1)得到化合物19a(3.5g)。LCMS(ESI-MS)m/z:396.1[M+H]+.
第二步2-((R)-3-(1-(1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-3-(三氟甲基)-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇19的合成
将化合物19a(1.0g,2.5mmol)和化合物1d(0.46g,2.5mmol)溶于乙腈(40mL)中，加入碳酸铯(4.1g,13mmol)，室温搅拌24小时。加水淬灭，乙酸乙酯萃取，有机相水洗，浓缩过flash柱子(DCM：MeOH＝10:1)得到化合物19(660mg)。LCMS(ESI-MS)m/z:544.28[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.58(s,1H),8.01(s,1H),7.84(s,1H),6.26(q,J＝7.0Hz,1H),4.19(s,2H),3.90(s,2H),3.53(s,2H),2.98–2.74(m,2H),2.56(d,J＝8.1Hz,2H),1.89(d,J＝7.1Hz,4H),1.80(s,1H),1.69(d,J＝13.4Hz,3H),1.53(s,2H),0.88(s,2H).
实施例20：((R)-3-(1-(1-(R)-1-(3,5-二氯吡啶-2-基)乙基)-3-(三氟甲基)-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(20)
化合物20的制备参照实施例19。LCMS(ESI)m/z:544.40[M+H]+。
1H NMR(400MHz,DMSO-d6)δ8.61(s,1H),8.27(s,1H),7.99(s,1H),6.33(q,J＝6.8Hz,1H),4.38(t,J＝5.6Hz,1H),4.16(s,2H),3.86(s,2H),3.49(t,J＝5.8Hz,2H),2.75(d,J＝10.6Hz,2H),2.55(dt,J＝9.0,4.8Hz,1H),2.35(t,J＝6.4Hz,2H),1.95–1.88(m,1H),1.85(d,J＝6.8Hz,3H),1.65(tt,J＝17.8,9.2Hz,4H),1.42(dd,J＝25.0,13.4Hz,1H),0.81(dd,J＝23.2,10.8Hz,1H).
实施例21：2-[(3R)-3-(1-{1-[(1R)-1-(4,6-二氯吡啶-3-基)乙基]-3-(3,3,3-三氟丙-1-炔基)吡唑并[4,3-b]吡嗪-6-基}氮杂环丁烷-3-基)六氢吡啶-1-基]乙-1-醇(21)
第一步2-[(3R)-3-(1-{1-[(1R)-1-(4,6-二氯吡啶-3-基)乙基]-3-(3,3,3-三氟丙-1-炔基)吡唑并[4,3-b]吡嗪-6-基}氮杂环丁烷-3-基)六氢吡啶-1-基]乙-1-醇(21)的合成
取一个单口瓶，加入TMEDA(350mg,3.0mmol)溶于DMF(2.0mL)，常温下依次加入碘化亚铜(570mg,3.0mmol)和碳酸钾(830mg,6.0mmol)。常温空气气氛下，剧烈搅拌20min。加入TMSCF3(570mg,4.0mmol)，常温空气气氛中，接着搅拌15min。反应变成深绿色。冷却到0℃，加入化合物1(1000mg,2.0mmol)和TMSCF3(570mg,4.0mmol)的DMF(2.0mL)混合溶液。0℃反应30min。升到室温反应6h。饱和氯化铵(10mL)淬灭，加入乙酸乙酯(50ml)，有机相水洗，无水硫酸钠干燥。减压浓缩。用制备HPLC纯化(碳酸氢铵体系制备)，得到化合物21(62mg)。LCMS(ESI)m/z:568.40[M+H]+。
1H NMR(400MHz,DMSO-d6)δ8.60(s,1H),7.98(s,1H),7.83(s,1H),6.26(q,J＝7.0Hz,1H),4.34(t,J＝5.4Hz,1H),4.18(s,2H),3.89(s,2H),3.46(q,J＝6.0Hz,2H),2.74-2.72(m,2H),2.56-2.54(m,1H),2.34(t,J＝6.4Hz,2H),1.93-1.87(m,4H),1.73-1.56(m,4H),1.42-1.39(m,1H),0.82-0.80(m,1H).
实施例22：2-[(3R)-3-{1-[3-(环丙基乙炔基)-1-[(1R)-1-(4,6-二氯吡啶-3-基)乙基]吡唑并[4,3-b]吡嗪-6-基]氮杂环丁烷-3-基}六氢吡啶-1-基]乙烷-1-醇(22)
化合物22的制备参照实施例1。LCMS(ESI)m/z:540.40[M+H]+。
1H NMR(400MHz,DMSO-d6)δ8.57(s,1H),7.85(d,J＝0.8Hz,1H),7.83(d,J＝0.8Hz,1H),6.18(q,J＝7.0Hz,1H),4.36(t,J＝5.4Hz,1H),4.21–4.12(m,2H),3.87(s,2H),3.48(q,J＝6.0Hz,2H),2.78–2.72(m,2H),2.59–2.53(m,1H),2.35(t,J＝6.4Hz,2H),1.91(t,J＝10.8Hz,1H),1.84(d,J＝7.2Hz,3H),1.78–1.57(m,6H),1.47–1.38(m,1H),0.95–0.90(m,2H),0.79(dd,J＝5.4,2.4Hz,2H).
实施例23：2-((R)-3-(1-(1-(R)-1-(3,5-二氯吡啶-2-基)乙基)-3-乙烯基-1H-吡唑并[4,3-b]吡啶-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(23)
第一步(R)-6-溴-1-(1-(3,5-二氯吡啶-2-基)乙基)-3-乙烯基-1H-吡唑并[4,3-b]吡啶(23c)
将化合物(R)-6-溴-1-(1-(3,5-二氯吡啶-2-基)乙基)-3-碘-1H-吡唑并[4,3-b]吡啶23a(500mg，1.0mmol)溶于1,4-二氧六环/水(5.0mL/1.0mL)中，室温下加入碳酸铯(650mg，2.0mmol)与甲磺酸[(4-(N,N-二甲氨基)苯基]二叔丁基膦(2-氨基-1,1-联苯-2-基)钯(250mg，0.35mmol)，氮气置换。冰浴加入乙烯 基硼酸频哪醇酯23b(170mg，1.1mmol)，室温搅拌过夜。用水淬灭，乙酸乙酯萃取，有机相干燥，过滤，滤液浓缩，硅胶柱纯化(石油醚：乙酸乙酯＝25:1)得到化合物23c(200mg)。LCMS(ESI-MS)m/z:397.14[M+H]+.
第四步2-((R)-3-(1-(1-(R)-1-(3,5-二氯吡啶-2-基)乙基)-3-乙烯基-1H-吡唑并[4,3-b]吡啶-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(23)
将化合物23c(150mg，0.38mmol)，化合物1d(320mg，1.74mmol)溶于N,N-二甲基甲酰胺(3.0mL)中，室温下加入碳酸铯(250mg，0.76mmol)与甲磺酸(4,5-双二苯基膦-9,9-二甲基氧杂蒽)(2-氨基-1,1'-联苯-2-基)钯(36mg，0.038mmol)，氮气置换。混合物100℃加热反应3小时。用水淬灭，乙酸乙酯萃取，有机相干燥，过滤，滤液浓缩，粗品用制备液相纯化(碳酸氢铵)得到化合物23(13mg)。LCMS(ESI-MS)m/z:501.40[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.68(d,J＝2.2Hz,1H),8.23(d,J＝2.2Hz,1H),7.92(d,J＝2.4Hz,1H),6.83–6.76(m,2H),6.58(dd,J＝17.8,2.6Hz,1H),6.25(d,J＝6.8Hz,1H),5.49(dd,J＝11.2,2.6Hz,1H),4.05–3.98(m,2H),3.67(td,J＝8.8,8.4,4.0Hz,2H),3.57(s,2H),2.93(s,2H),2.62–2.53(m,2H),1.91–1.83(m,1H),1.80(d,J＝6.8Hz,3H),1.78–1.61(m,4H),1.60–1.44(m,2H),1.02–0.81(m,2H).
实施例24：1-(((R)-1-(3,5-二氯吡啶-2-基)乙基)-6-(3-(((R)-1-(2-羟乙基)哌啶-3-基)氮杂环丁烷-1-基)-1H-吡唑并[4,3-b]吡啶-3-腈(24)
第一步(R)-6-溴-1-(1-(3,5-二氯吡啶-2-基)乙基)-1H-吡唑并[4,3-b]吡啶-3-腈(24a)
将化合物23a(500mg，1.0mmol)溶于N,N-二甲基乙酰胺(6.0mL)，室温下加氰化锌(61mg，0.52mmol)，三(二亚苄基丙酮)二钯(81mg，0.10mmol)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(58mg，0.10mmol)与四甲基乙二胺(35mg，0.30mmol)，氮气置换。90℃加热反应1小时。加水淬灭，乙酸乙酯萃取，有机相干燥过滤，滤液浓缩，硅胶柱纯化(石油醚：乙酸乙酯＝25:1)得化合物24a(220mg)。LCMS(ESI-MS)m/z:396.05[M+H]+.
第二步1-(((R)-1-(3,5-二氯吡啶-2-基)乙基)-6-(3-(((R)-1-(2-羟乙基)哌啶-3-基)氮杂环丁烷-1-基)-1H-吡唑并[4,3-b]吡啶-3-腈(24)
将化合物24a(150mg，0.38mmol)，1d(320mg，1.7mmol)溶于甲苯(1.2mL)中，室温下加入碳酸铯(190mg，0.57mmol)，(2-二环己膦-2',6'-二异丙氧基-1,1'-联苯)[2-(2-氨基乙基苯基)]氯化钯甲基叔丁基醚络合物(31mg，0.038mmol)与2-二环己基磷-2',6'-二异丙氧基-1,1'-联苯(18mg，0.038mmol)，氮气置换。混合物100℃加热反应3小时。用水淬灭，乙酸乙酯萃取，有机相干燥，过滤，滤液浓缩，粗品用制备液相纯化(氨水)得到化合物24(4.2mg)。LCMS(ESI-MS)m/z:500.40[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.68(d,J＝2.2Hz,1H),8.30(d,J＝2.2Hz,1H),8.09(d,J＝2.2Hz,1H),6.94(t,J＝2.0Hz,1H),6.45(d,J＝6.8Hz,1H),4.37(s,1H),4.08–4.03(m,2H),3.73(d,J＝8.4Hz,2H),3.49 (t,J＝6.0Hz,2H),2.80–2.74(m,2H),2.57(d,J＝7.8Hz,1H),2.39–2.34(m,2H),1.85(d,J＝6.8Hz,3H),1.72–1.62(m,4H),1.48–1.40(m,2H),0.86(d,J＝5.4Hz,1H).
实施例25：1-[(1R)-1-(3,5-二氯吡啶-2-基)乙基]-6-{3-[(3R)-1-[(2-羟乙基)六氢吡啶-3-基]氮杂环丁烷-1-基}吡咯并[3,2-b]吡啶-3-腈(25)
第一步6-溴-3-碘-1H-吡咯并[3,2-b]吡啶25b的合成
取50mL烧瓶，加入6-溴-1H-吡咯并[3,2-b]吡啶25a(5.0g,25mmol)，DMF(25mL)，冰浴搅拌10min，加入氢氧化钾(1.1g,20mmol)，碘(2.6g,10mmol)，升至室温反应5小时，冰浴下加入饱和亚硫酸氢钠水溶液淬灭反应，乙酸乙酯萃取(400mL*3)，饱和氯化钠水溶液(200mL)洗涤，无水硫酸钠干燥，旋干溶剂，得化合物25b(6.2g)。LCMS(ESI-MS)m/z:323.0[M+H]+.
第二步6-溴-1-[(1R)-1-(3,5-二氯吡啶-2-基)乙基]-3-碘代吡咯并[3,2-b]吡啶25d的合成
取25mL三口烧瓶，加入化合物25b(1.1g,3.3mmol)，二氯甲烷(6.0mL)，(1R)-1-(3,5-二氯吡啶-2-基)乙烷-1-醇25c(0.63g,3.3mmol)，三苯基磷(1.4g,5.3mmol)，氮气保护，冰浴搅拌反应10min，加入DIAD(1.1g,5.6mmol)，继续搅拌反应2小时后倒入水中，DCM萃取三次(20mL*3)，饱和氯化钠水溶液洗涤(30mL)，无水硫酸钠干燥，Flash柱纯化，得化合物25d(1.6g)。LCMS(ESI-MS)m/z:496.04[M+H]+.
第三步6-溴-1-[(1R)-1-(3,5-二氯吡啶-2-基)乙基]吡咯并[3,2-b]吡啶-3-腈25e的合成
取100mL烧瓶，加入25d(1200mg,2.4mmol)，DMA(10mL)，Zn(CN)2(140mg,1.2mmol)，Pd2(dba)3(220mg,0.24mmol)，XANTPHOS(140mg,0.24mmol)，2,5-二甲基-2,5-二氮杂己烷(83mg,0.71mmol)，氮气保护，70℃搅拌反应2h，LCMS监测，原料反应完全，将反应液倒入水(60mL)中，加入饱和NaHCO3水溶液(20mL)，加入EDTA(0.50g)，EA萃取三次(30mL*3)，饱和氯化钠水溶液洗涤(60mL)，无水硫酸钠干燥，Flash柱纯化，得化合物25e(390mg)。LCMS(ESI-MS)m/z:395.13[M+H]+.
第四步1-[(1R)-1-(3,5-二氯吡啶-2-基)乙基]-6-{3-[(3R)-1-[(2-羟乙基)六氢吡啶-3-基]氮杂环丁烷-1-基}吡咯并[3,2-b]吡啶-3-腈25的合成
取75mL封管，加入化合物25e(220mg,1.2mmol)，叔丁醇(15mL)，EPhos Pd G4(110mg,0.12mmol)，EPhos(65mg,0.12mmol)，碳酸铯(390mg,1.2mmol)，氮气保护，85℃加热反应6小时，LCMS监测，原料反应完全，硅藻土过滤，旋干溶剂，HPLC制备，得化合物25(16mg)。LCMS(ESI-MS)m/z:250.20[M/2+H]+.
1H NMR(400MHz,DMSO-d6)δ8.63(d,J＝2.1Hz,1H),8.37(s,1H),8.30(d,J＝2.2Hz,1H),7.82(d,J＝2.3Hz,1H),6.80(d,J＝2.4Hz,1H),6.21(q,J＝6.8Hz,1H),4.36(s,1H),3.93(dt,J＝13.3,7.6Hz,2H),3.56(q,J＝7.1Hz,2H),3.48(t,J＝6.4Hz,2H),2.75(t,J＝6.9Hz,2H),2.36(t,J＝6.3Hz,2H),1.91(dt,J＝11.8,6.5Hz,1H),1.83(d,J＝6.8Hz,3H),1.76–1.54(m,5H),1.42(d,J＝12.3Hz,1H),0.84(dd,J＝17.2,7.6Hz,1H).
实施例26:5-[(1R)-1-(4,6-二氯吡啶-3-基)乙基]-3-{3-[(3R)-1-(2-羟乙基)六氢吡啶-3-基]氮杂环丁烷-1-基}-7-(三氟甲基)吡咯并[3,2-b]吡嗪-6-甲腈(26)
第一步3-氯-5-[(4-甲基苯基)二氧代-λ6-硫基]吡咯并[3,2-b]吡嗪26b的合成
取一个单口瓶，将化合物26a(5.0g,33mmol)溶于DCM(50mL)，常温加入三乙胺(23mL，160mmol)，对甲苯磺酰氯(10g,39mmol)，DMAP(0.80g,6.5mmol)。常温反应过夜。加水，DCM萃取，无水硫酸钠干燥，过滤，减压浓缩。硅胶柱(乙酸乙酯：石油醚＝10-30％)纯化，得化合物26b(9.0g)。LCMS(ESI-MS)m/z＝308.14[M+H]+
第二步3-氯-5-[(4-甲基苯基)二氧代-λ6-硫基]吡咯并[3,2-b]吡嗪-6-甲腈24c的合成
取一个三口瓶，将化合物26b(1.5g,4.9mmol)溶于四氢呋喃(5.0mL)，常温氮气置换三次，干冰乙酸乙酯浴下，滴加正丁基锂(2.9mL,7.3mmol)，反应1h，滴加4-甲苯磺酰氰(1.8g,9.7mmol)的四氢呋喃(5.0mL)溶液。相同温度反应30min，常温反应过夜。加饱和氯化铵水溶液，乙酸乙酯萃取三次，无水硫酸钠干燥，过滤，减压浓缩。硅胶柱(乙酸乙酯：石油醚＝10-30％纯化)，得化合物26c(410mg)。LCMS(ESI-MS)m/z＝333.14[M+H]+
第三步3-氯-5H-吡咯并[3,2-b]吡嗪-6-甲腈26d的合成
取一个单口瓶，依次加入化合物26c(2.7g,8.2mmol)，悬浮在甲醇(30mL)，常温加入碳酸钾(2.3g,16mmol)，50℃下反应1h。变透明澄清减压蒸去溶剂。加水，用乙酸乙酯萃取，无水硫酸钠干燥，过滤，减压浓缩。得到化合物26d(1.7g,9.6mmol)，直接用作下一步。LCMS(ESI-MS)m/z＝178.99[M+H]+
第四步3-氯-7-碘-5H-吡咯并[3,2-b]吡嗪-6-甲腈26e的合成
取一个单口瓶，依次加入化合物26d(1.7g,9.6mmol)，溶解在DMF(30mL)，分批加入氢氧化钾(2.0g,35mmol)，常温下反应30min。冰水浴条件下加入碘(5.0g，19mmol)的DMF(1.0mL)。常温反应20h。加入饱和亚硫酸钠水溶液，淬灭过量的碘，用乙酸乙酯萃取三次，合并有机相，无水硫酸钠干燥，减压蒸去溶剂。得到化合物26e(810mg)，直接用作下一步。LCMS(ESI-MS)m/z＝304.98[M+H]+
第五步3-氯-5-[(1R)-1-(4,6-二氯吡啶-3-基)乙基]-7-碘吡咯并[3,2-b]吡嗪-6-甲腈26f的合成
取一个三口瓶，加入化合物1h(190mg,1.0mmol)，化合物26e(300mg,1.0mmol)，常温下氮气置换，冰水浴下加入三苯基磷(520mg,2.0mmol)的DCM(1.0mL)溶液，同时加入DIAD(500mg,2.5mmol)的DCM(1.0mL)溶液，相同温度反应2h。加入甲醇，减压蒸去溶剂。硅胶柱(乙酸乙酯：石油醚＝10％)纯化，得26f(380mg)。LCMS(ESI-MS)m/z＝478.07[M+H]+
第六步3-氯-5-[(1R)-1-(4,6-二氯吡啶-3-基)乙基]-7-(三氟甲基)吡咯并[3,2-b]吡嗪-6-甲腈26g的合成
取一个单口瓶，依次加入化合物26f(160mg,0.33mmol)，溶于DMF(5.0mL)，加入碘化亚铜(130mg,0.40mmol)，六甲基磷酰三胺(0.070mL,0.40mmol)，氟磺酰基二氟乙酸甲酯(260mg,1.3mmol)，常温下氮气置换，100℃反应2h。冷却到常温，加饱和氯化钠水溶液，乙酸乙酯萃取三次，合并有机相，无水硫酸钠干燥，减压蒸去溶剂。硅胶柱(乙酸乙酯：石油醚＝10％)纯化，得化合物26g(95mg)。LCMS(ESI-MS)m/z＝420.01[M+H]+
第七步5-[(1R)-1-(4,6-二氯吡啶-3-基)乙基]-3-{3-[(3R)-1-(2-羟乙基)六氢吡啶-3-基]氮杂环丁烷-1-基}-7-(三氟甲基)吡咯并[3,2-b]吡嗪-6-甲腈26的合成
取一个单口瓶，依次加入化合物26g(90mg,0.21mmol)，化合物1d(590mg，0.6mmol)，溶解在乙腈(10mL)，加入碳酸铯(700mg,2.1mmol)，常温下反应2h。加入水溶液，二氯甲烷萃取三次，合并有机相，无水硫酸钠干燥，减压蒸去溶剂。用制备HPLC纯化，得到化合物26(48mg)。LCMS(ESI-MS)m/z＝568.40[M+H]+
1H NMR(400MHz,DMSO-d6)δ8.94(s,1H),7.98(d,J＝1.5Hz,1H),7.80(d,J＝2.2Hz,1H),6.06-6.01(m,1H),4.59-4.33(m,1H),4.20-3.95(m,2H),3.87(t,J＝7.4Hz,1H),3.69(s,1H),3.49-3.44(m,2H),2.80-2.70(m,2H),2.60-2.50(m,1H),2.40-2.30(m,2H),2.02(d,J＝7.1Hz,3H),1.90(t,J＝11.1Hz,1H),1.75–1.54(m,4H),1.50-1.35(m,1H),0.90-0.75(m,1H).
实施例27：2-[(3R)-3-{1-[3-(环丙基乙炔基)-1-[1-(2,4-二氯嘧啶-5-基)乙基]吡唑并[4,3-b]吡嗪-6-基]氮杂环丁烷-3-基}六氢吡啶-1-基]乙烷-1-醇(27)
化合物27的制备参照实施例1。LCMS(ESI-MS)m/z:541.40[M+H]+.
1H NMR(400MHz,DMSO-d6)δ9.02(d,J＝2.3Hz,1H),7.86(s,1H),6.11(q,J＝7.0Hz,1H),4.41(s,1H),4.17(s,2H),3.88(s,2H),3.51(s,2H),2.80(s,2H),2.52(s,2H),2.41(s,1H),1.99(s,1H),1.85(d,J＝7.0Hz,3H),1.78–1.56(m,4H),1.46(s,1H),0.93(dq,J＝6.5,3.9,3.5Hz,2H),0.86(s,1H),0.78(dq,J＝7.0,4.0Hz,2H).
实施例28：2-[(3R)-3-(1-{3-[(1E)-2-环丙基乙烯基]-1-[(1R)-1-(4,6-二氯吡啶-3-基)乙基]吡唑并[4,3-b]吡嗪-6-基}氮杂环丁烷-3-基)六氢吡啶-1-基]乙烷-1-醇(28)
化合物28的制备参照实施例23。LCMS(ESI-MS)m/z:542.4[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.56(d,J＝2.4Hz,1H),7.81(d,J＝5.3Hz,2H),6.59(d,J＝4.4Hz,2H),6.17(q,J＝7.1Hz,1H),4.40(s,1H),4.14(t,J＝8.4Hz,2H),3.84(q,J＝6.6Hz,2H),3.49(t,J＝6.4Hz,2H),2.76(d,J＝10.6Hz,2H),2.36(t,J＝6.2Hz,2H),1.96–1.82(m,4H),1.82–1.51(m,6H),1.44(t,J＝12.0Hz,1H),0.89–0.76(m,3H),0.55(d,J＝4.8Hz,2H).
实施例29：5-[(1R)-1-(4,6-二氯吡啶-3-基)乙基]-3-{3-[(3R)-1-(2-羟乙基)六氢吡啶-3-基]氮杂环丁烷-1-基}吡咯并[3,2-b]吡嗪-6.7-二甲腈(29)
化合物29的制备参照实施例26.LCMS(ESI-MS)m/z＝525.38[M+H]+
1H NMR(400MHz,DMSO-d6)δ8.92(s,1H),8.01(s,1H),7.82(s,1H),6.08-6.02(m,1H),4.34(t,J＝5.4Hz,1H),4.20-4.00(m,2H),3.92-3.84(m,1H),3.80-3.60(m,1H),3.47(q,J＝6.1Hz,2H),2.78–2.69(m,2H),2.52-2.60(m,1H),2.34(dd,J＝7.2,5.6Hz,2H),2.01(d,J＝7.0Hz,3H),1.96-1.86(m,1H),1.75-1.55(m,4H),1.47-1.37(m,1H),0.90-0.75(m,1H).
实施例30：1-(1-(4,6-二氯吡啶-3-基)乙基)-6-(3-((R)-1-(2-羟乙基)哌啶-3-基)氮杂环丁烷-1-基)-1H-吡唑并[4,3-b]吡啶-3-甲腈(30)
化合物30的制备参照实施例24。LCMS(ESI-MS)m/z:500.40[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.59(d,J＝1.7Hz,1H),8.09(d,J＝2.2Hz,1H),7.85(s,1H),7.04(d,J＝2.3Hz,1H),6.36(q,J＝6.8,6.3Hz,1H),4.35(t,J＝4.8Hz,1H),4.06(p,J＝7.4Hz,2H),3.72(s,2H),3.47(q,J＝5.9Hz,2H),2.75(d,J＝10.5Hz,2H),2.56(s,1H),2.34(t,J＝6.3Hz,2H),1.97(dd,J＝16.6,10.0Hz,1H),1.88(d,J＝6.8Hz,3H),1.64(dt,J＝25.6,12.8Hz,4H),1.42(d,J＝9.4Hz,1H),0.87–0.80(m,1H).
实施例31：2-((R)-2-(1-(3-(环丙基乙炔基)-1-(R)-1-(4,6-二氯吡啶-3-基)乙基)-1H-吡唑并[3,4-b]吡嗪-6-基)氮杂环丁烷-3-基)吗啉基)乙烷-1-醇(31)
化合物31的制备参照实施例1。LCMS(ESI-MS)m/z:542.40[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.54(d,J＝1.3Hz,1H),7.83(d,J＝12.5Hz,2H),6.17(q,J＝7.0Hz,1H),4.12(s,2H),3.97(d,J＝27.7Hz,2H),3.78(d,J＝11.6Hz,1H),3.59(td,J＝7.4,7.0,3.7Hz,1H),3.55–3.45(m,3H),2.76(d,J＝11.8Hz,2H),2.69(d,J＝11.3Hz,1H),2.37(t,J＝6.2Hz,3H),2.03(dd,J＝11.6,8.4Hz,1H),1.83(d,J＝7.1Hz,3H),1.72(td,J＝10.7,3.1Hz,1H),1.64–1.57(m,1H),0.90(dt,J＝8.1,3.2Hz,2H),0.81–0.72(m,2H).
实施例32：2-[2-(1-{1-[1-(4,6-二氯吡啶-3-基)乙基]-3-(三氟甲基)吡唑并[4,3-b]吡嗪-6-基}氮杂环丁烷-3-基)-1,4-恶嗪-4-基]乙烷-1-醇(32)
化合物32的制备参照实施例19。LCMS(ESI-MS)m/z:546.35[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.56(d,J＝1.8Hz,1H),8.01(s,1H),7.83(s,1H),6.26(q,J＝7.0Hz,1H),4.27–3.91(m,4H),3.78(d,J＝11.0Hz,1H),3.63–3.47(m,5H),2.89–2.63(m,3H),2.37(t,J＝6.2Hz,2H),2.14–1.61(m,5H).
实施例33：1-[1-(4,6-二氯吡啶-3-基)乙基]-6-{3-[(3R)-1-(2-羟乙基)六氢吡啶-3-基]氮杂环丁烷-1-基}吡咯并[3,2-b]吡啶-3-腈(33)
化合物33的制备参照实施例25。LCMS(ESI-MS)m/z:250.20[1/2M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.53(d,J＝2.8Hz,1H),8.36(d,J＝2.8Hz,1H),7.95–7.79(m,2H),6.90(s,1H),6.09(d,J＝6.9Hz,1H),4.38(s,1H),3.95(dq,J＝8.0,4.0Hz,2H),3.59(q,J＝7.2Hz,2H),3.49(t,J＝6.3Hz,2H),2.76(d,J＝10.1Hz,2H),2.36(t,J＝6.4Hz,2H),1.90(d,J＝7.2Hz,4H),1.65(dq,J＝23.5,12.2,11.4Hz,5H),1.44(t,J＝12.3Hz,1H),0.83(d,J＝11.9Hz,1H).
实施例34：2-((3R)-3-(1-(1-(4-(4,6-二氯吡啶-3-基)乙基)-3-(三氟甲基)-1H-吡唑并[4,3-b]吡啶-6-基)氮杂环丁烷-3-基)哌啶-1-基)乙烷-1-醇(34)
化合物34的制备参照实施例19。LCMS(ESI-MS)m/z:543.39[M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.55(d,J＝1.8Hz,1H),8.06(d,J＝2.2Hz,1H),7.85(d,J＝1.6Hz,1H),6.97(d,J＝2.4Hz,1H),6.32(q,J＝6.8Hz,1H),4.37(s,1H),4.06(h,J＝4.6,3.8Hz,2H),3.73(d,J＝8.6Hz,2H),3.49(t,J＝8.2Hz,2H),2.81–2.74(m,2H),2.61–2.55(m,1H),2.38(t,J＝6.2Hz,2H),1.97(d,J＝10.4Hz,1H),1.90(d,J＝6.8Hz,3H),1.75–1.60(m,4H),1.44(d,J＝12.4Hz,1H),0.86(dd,J＝22.4,10.6Hz,1H).
实施例35：2-[(3R)-3-{1-[3-(环丙基乙炔基)-1-[1-(4,6-二氯吡啶-3-基)乙基]吡唑并[4,3-c][1,2]二嗪-6-基]氮杂环丁烷-3-基}六氢吡啶-1-基]乙烷-1-醇(35)
第一步6-氯-4-二嗪基-1,2-二嗪-3-羧酸甲酯35b的合成
取100mL烧瓶，加入4,6-二氯-1,2-二氮-3-羧酸甲酯35a(10g,48mmol)，乙醇(100mL)，氮气保护，冰浴下缓慢加入水合肼(7.3mL,120mmol)，5min加完，保持冰浴反应，5min后有大量固体析出，保持温度反应1小时，LCMS监测，原料基本反应完全，过滤，水洗滤饼，旋干滤饼，加入DCM/EA(1:3，300mL)溶解滤饼，滤饼溶解不完全，有大量悬浮物，加入硅胶拌样，Flash柱纯化得化合物35b(4.9g)。LCMS(ESI-MS)m/z:203.06[M+H]+.
第二步6-氯-1H-吡唑并[4,3-c][1,2]二嗪35c的合成
取250mL三口烧瓶，加入化合物35b(3.5g,17mmol)，DCM(30mL)，插入低温温度计，氮气保护，干冰-乙醇浴降温至-70℃以下，缓慢加入DIBAL-H(17mL,26mmol)，保持温度在-70℃以下，滴毕，缓慢升至室温反应4小时，LCMS监测，有少量原料没有反应完全，加入H2O(2.0mL)，乙醇(50mL)，直接旋干溶剂，加入硅胶拌样，Flash柱纯化得化合物35c(1.4g)。LCMS(ESI-MS)m/z:155.04[M+H]+.
第三步6-氯-3-碘-1H-吡唑并[4,3-c][1,2]二嗪35d的合成
取100mL烧瓶，加入化合物35c(1.4g,8.7mmol)，DMF(15mL)，NIS(3.9g,17mmol)，氮气保护，70℃加热反应1小时，LCMS监测，反应完全，将反应液直接冻干，向残余物中加入乙酸乙酯(50mL)，石油醚(30mL)，搅拌打浆1小时，过滤，将不溶物旋干，直接用于下步反应。LCMS(ESI-MS)m/z:281.01[M+H]+.
第四步6-氯-1-(5,5-二甲基-2-氧-5-硅己基-1-基)-3-碘代吡唑并[4,3-][1,2]二嗪35e的合成
取100mL烧瓶，加入化合物35d(2.2g,7.7mmol)，THF(40mL)，碳酸铯(5.0g,15mmol)，SEMCl(1.7g,10mmol)，氮气保护，室温搅拌过夜，LCMS监测，原料反应完全，将反应液倒入水中(100mL)，乙酸乙酯萃取三次(30mL*3)，饱和氯化钠水溶液洗涤(100mL)，无水硫酸钠干燥，过滤，旋干溶剂，加入硅胶拌样，Flash柱纯化得化合物35e(1.3g)。LCMS(ESI-MS)m/z:411.13[M+H]+.
第五步2-[(3R)-3-{1-[1-(5,5-二甲基-2-氧-5-硅己基-1-基)-3-碘代吡唑并[4,3-][1,2]二嗪-6-基]氮杂环丁烷-3-基}六氢吡啶-1-基]乙烷-1-醇35f的合成
取15mL封管，加入化合物35e(410mg,1.0mmol)，DMSO(5.0mL)，化合物1d(280mg,1.5mmol)，DIEA(390mg,3.0mmol)，氮气保护，120℃加热反应2小时，LCMS监测，原料基本反应完全，将反应液倒入水(50mL)中，乙酸乙酯萃取三次(10mL*3)，饱和氯化钠水溶液洗涤(20mL)，无水硫酸钠干 燥，过滤，加入硅胶拌样，Flash柱纯化得化合物35f(140mg)。LCMS(ESI-MS)m/z:280.18[1/2M+H]+.
第六步2-[(3R)-3-{1-[3-(环丙基乙炔基)-1-(5,5-二甲基-2-氧-5-硅己基-1-基)吡唑并[4,3-][1,2]二嗪-6-基]氮杂环丁烷-3-基}六氢吡啶-1-基]乙烷-1-醇35g的合成
取50mL烧瓶，加入化合物35f(200mg,0.36mmol)，THF(5.0mL)，双三苯基磷二氯化钯(50mg,0.072mmol)，CuI(20mg,0.10mmol)，TEA(0.25mL,1.8mmol)，乙炔基环丙烷(71mg,1.1mmol)，氮气下45℃加热反应1h，LCMS监测，基本反应完全，倒入水(20mL)中，乙酸乙酯萃取三次(10mL*3)，饱和氯化钠溶液洗涤(20mL)，无水硫酸钠干燥，过滤，Flash柱纯化得得化合物35g(140mg)。LCMS(ESI-MS)m/z:249.29[1/2M+H+].
第七步2-[(3R)-3-{1-[3-(环丙基乙炔基)-1H-吡唑并[4,3-][1,2]二嗪-6-基]氮杂环丁烷-3-基}六氢吡啶-1-基]乙烷-1-醇35h的合成
取15mL封管，加化合物35g(110mg,0.22mmol)，TBAF的THF溶液(5.0mL,5.0mmol)，2,5-二甲基-2,5-二氮杂己烷(130mg,1.1mmol)，氮气下80℃反应5h，LCMS监测，基本反应完全，倒入饱和氯化钠溶液洗涤四次(15mL)，无水硫酸钠干燥，旋干溶剂，得化合物35h粗品，直接用于下步反应。
LCMS(ESI-MS)m/z:184.17[1/2M+H]+.
第八步2-[(3R)-3-{1-[3-(环丙基乙炔基)-1-[1-(4,6-二氯吡啶-3-基)乙基]吡唑并[4,3-c][1,2]二嗪-6-基]氮杂环丁烷-3-基}六氢吡啶-1-基]乙烷-1-醇35的合成
取15mL封管，加入化合物35h(60mg,0.16mmol)，DMF(2.0mL)，5-(1-溴乙基)-2,4-二氯吡啶35i(130mg,0.49mmol)，碳酸钾(91mg,0.66mmol)，氮气保护，室温搅拌反应过夜，LCMS监测，原料基本反应完全，将反应液倒入水(30mL)中，EA萃取三次(15mL*3)，饱和氯化钠水溶液洗涤(20mL)，无水硫酸钠干燥，过滤，旋干溶剂，HPLC纯化，冻干得化合物35(20mg)。LCMS(ESI-MS)m/z:270.76[1/2M+H]+.
1H NMR(400MHz,DMSO-d6)δ8.60(s,1H),7.83(d,J＝2.0Hz,1H),6.80(d,J＝1.8Hz,1H),6.16(q,J＝6.8Hz,1H),4.37(t,J＝5.4Hz,1H),4.14(q,J＝7.4Hz,2H),3.82(t,J＝7.2Hz,2H),3.49(q,J＝5.9Hz,2H),2.84–2.71(m,2H),2.59(s,1H),2.37(t,J＝6.4Hz,2H),1.93(t,J＝11.1Hz,1H),1.83(d,J＝6.9Hz,3H),1.79–1.55(m,5H),1.44(d,J＝12.2Hz,1H),1.00–0.92(m,2H),0.91–0.78(m,3H).
生物活性试验
实验一、本发明CCR4抑制剂小分子化合物的体外活性测定
1.实验目的
本实验通过高通量实时荧光分析检测(FLIPR)方法测试本发明化合物对CCR4受体的抑制作用，根据半抑制浓度(IC50)评价化合物的体外活性。
2.实验方法
2.1实验材料
表1实验材料
2.2实验流程
(1)使用Echo将测试化合物在100％DMSO中连续稀释3倍至10份。然后将900nL化合物转移到目标板上。向化合物板中加入30μL测定缓冲液。
(2)从培养箱中取出细胞板，丢弃培养基，加入20μL分析缓冲液。使用移液管将20μL 2X Fluo-4 DirectTM无冲洗加载缓冲液轻轻地分配到384孔细胞培养板中。
(3)然后将10μL参考物和化合物从化合物板转移到细胞板。细胞板中的最终体积为50μL。
(4)启动FLIPRTETRA
(5)将10μL 6×EC80浓度的激动剂参考化合物转移至细胞板。
(6)读取荧光信号
(7)对于拮抗剂试验，1相加计算从读数1到允许的最大值的“最大值-最小值”，1相加从读数1开始计算“最大值”到允许的130。
(8)分析数据
2.3实验结果
表1本发明化合物体外活性实验结果
结论：本发明化合物对CCR4受体具有明显的抑制活性。
实验二、本发明化合物的小鼠体内药效实验
1.造模及给药
8周龄雌性免疫缺陷小鼠，平均体重20g。小鼠适应3天后开始实验，实验分为四组，模型组、FLX475组和实施例1化合物组，每组8只。实验前培养人源化肝癌细胞Hepg2，培养至一定数量后，每只皮下注射1*107个细胞，待肿瘤长至一定大小后，模型组、FLX475组和实施例化合物35组按10mL/kg灌胃及注射给药，每日一次；模型组灌胃/注射等量溶媒，连续28d。具体设计见表3。
表3化合物体内药效实验方案
2.实验动物
雌性免疫缺陷小鼠，8周龄。小鼠均购自北京维通利华实验动物技术有限公司，许可证号SCXK(沪)2017-0011，动物合格证编号：20170011003865。
3.实验药物
实施例化合物35。
4.药物的配置
称取适量化合物，溶于30％聚乙二醇(PEG400)涡旋超声，加入70％生理盐水3mg/mL的澄清透明溶液。
5.给药
具体给药按照给药方案进行。
6.检测指标和方法
每天观察小鼠肿瘤大小及体重，一周三次测量肿瘤大小，终点解剖，取肿瘤，拍照记录终点肿瘤大小。
7.实验结果及结论
经过人源化Hepg2肿瘤细胞移植后，模型对照组小鼠肿瘤大小随天数持续增高，实施例化合物35第14天后肿瘤体积开始明显减小。
实验三、本发明化合物对hERG通道电流的影响
1.实验目的
通过手动膜片钳技术检测化合物对hERG通道电流的影响，评估本发明化合物的心脏安全性。
2.实验方法
2.1实验材料
2.2实验流程
1)hERG钾通道稳定表达的HEK-293细胞系在含有10％胎牛血清及0.8mg/mL G418的DMEM培养基中培养，培养温度为37℃，二氧化碳浓度为5％。
2)细胞传代：除去旧培养基并用PBS洗一次，然后加入1mL TrypLETMExpress溶液，37℃孵育0.5min左右。当细胞从皿底脱离，加入约5mL 37℃预热的完全培养基。将细胞悬液用吸管轻轻吹打使聚集的细胞分离。将细胞悬液转移至无菌的离心管中，1000rpm离心5min收集细胞。扩增或维持培养，将细胞接种于6cm细胞培养皿，每个细胞培养皿接种细胞量为2.5×105细胞(最终体积：5mL)。
3)为维持细胞的电生理活性，细胞密度必须不能超过80％。
4)膜片钳检测，试验之前细胞用TrypLETMExpress分离，将4×103细胞铺到盖玻片上，在24孔板中培养(最终体积：500μL)，18个小时后，进行试验检测。全细胞膜片钳记录hERG钾电流的电压刺激方案如下：当形成全细胞封接后细胞膜电压钳制于-80mV。钳制电压由-80mV除极至-50mV维持0.5s(作为漏电流检测)，然后阶跃至30mV维持2.5s，再迅速恢复至-50mV维持4s可以激发出hERG通道的尾电流。每隔10s重复采集数据，观察药物对hERG尾电流的作用。以0.5s的-50mV刺激作为漏电流检测。试验数据由IPA放大器(Sutter Instrument)进行采集并储存于Sutter Patch软件中。
当全细胞记录的hERG电流稳定后开始给药，每个药物浓度作用至5min(或者电流至稳定)后检测下一个浓度，每一个测试化合物检测一个或多个浓度。将铺有细胞的盖玻片置于倒置显微镜中的记录浴槽中，测试化合物工作液以及不含化合物的外液利用重力灌流给药的方式从低浓度到高浓度依次流经记录浴槽从而作用于细胞，同时在记录中利用蠕动泵进行液体置换。每一个细胞在不含化合物的外液中检测到的电流作为自己的对照组。每个浓度至少使用两个细胞独立重复检测两次。所有电生理试验在室温下进行。
2.3数据分析
首先将每一个药物浓度作用后的尾电流(Peak tail currentcompound)和空白溶媒处理组对照尾电流(Peak tail currentControl)归一化(Normalized)(Peak tail currentcompound/Peak tail currentControl)，然后计算每一个药物浓度对应的抑制率(1-(Peak tail currentcompound/Peak tail currentControl)。并对每一个浓 度抑制率计算平均数(Mean)，标准差(SD)和标准误(SE)，数据以Mean±SE表示并保存于Excel。Y＝1/(1+10^((LogIC50-X)*HillSlope)，用以上方程计算每种化合物的IC50值，并对浓度效应曲线进行非线性拟合，其中IC50为半抑制浓度。IC50的计算以及曲线拟合利用GraphPad Prism软件完成。
2.4实验结果
其中，D<0.1，0.1≤C＜1.0，1.0≤B＜1.5，1.5≤A。
3.实验结论
与阳性参照物FLX-475相比，本发明化合物心脏安全性指标hERG毒性有所改善。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本请所附权利要求书。

Claims (19)

  1. 式(Ⅰ)所示的化合物、其立体异构体、互变异构体或其可药用盐，
    其特征在于，环A为4-8元芳杂环；
    X1或X2各自独立地为N或CH；
    X3为N或CRa；
    Y为-O-或-CH2-；
    Ra为-H、-C1-3烷基、-卤素或-CN，所述-C1-3烷基任选地被一个或化合价所允许范围内的多个卤素所取代；
    L1为键、-C1-3亚烷基-、-C2-4亚烯基-或-C2-4亚炔基-；
    R1为-C1-3亚烷基-OH；
    R2为-H、-CN、-C1-5烷基、3-8元环烷基或3-8元杂环烷基，所述-C1-5烷基任选地被一个或化合价所允许范围内的多个卤素取代，所述3-8元环烷基或3-8元杂环烷基任选地被一个或者化合价所允许范围内的多个-C1-5烷基或-卤素所取代；
    R3为-C1-5烷基；
    R4各自独立地为卤素；
    n为0、1、2或3。
  2. 根据权利要求1所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，环A为5-6元芳杂环；优选地，所述5-6元芳杂环的杂原子为N、O或S。
  3. 根据权利要求1或2所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，环A为其中E1、E2各自独立地为CH、N、O或S，E3为键、CH或N，且E1、E2、E3不同时为CH；
    优选地，环A为
  4. 根据权利要求3所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，E1为S、N、O，E2为CH、N、O或S，E3为键；
    优选地，E1为O，E2为N，E3为键；或E1为N，E2为O，E3为键；或E1为S，E2为N，E3为键；或E1为N，E2为S，E3为键；或E1为S，E2为CH，E3为键。
  5. 根据权利要求3所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，E1、E2、E3各自独立地为CH或N，且E1、E2、E3不同时为CH；
    优选地，E1为CH，E2为N，E3为CH；或E1为CH，E2为CH，E3为N；或E1为N，E2为N，E3为CH；E1为CH，E2为N，E3为N；或E1为N，E2为N，E3为CH；E1为N，E2为N，E3为N。
  6. 根据权利要求1-3任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，环A为
    优选地，环A为
  7. 根据权利要求1-6任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，n为0、1或2，优选为2。
  8. 根据权利要求1-7任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，X1或X2不同时为N；
    优选地，X1为CH，X2为N，X3为N；或X1为CH，X2为N，X3为CRa；或X1为N，X2为CH，X3为N；或X1为CH，X2为CH，X3为N；或X1为CH，X2为CH，X3为CRa；
    更优选地，Ra为-H或-CN。
  9. 根据权利要求1-8任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，L1为键、-C2-4亚烯基-或-C2-4亚炔基-，优选为键、-C≡C-或-CH＝CH-；和/或
    R1为-CH2CH2OH。
  10. 根据权利要求1-9任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，R2为-H、-CN、-C1-5烷基、3-5元环烷基或3-5元杂环烷基，所述-C1-5烷基任选地被一个或化合价所允许范围内的多个卤素取代，所述3-5元环烷基或3-5元杂环烷基任选地被一个或者化合价所允许范围内的多个-C1-3烷基或-卤素所取代；
    优选地，所述-C1-5烷基为甲基或叔丁基，所述3-5元环烷基为环丙基，所述3-5元杂环烷基为氧杂环丁基或氮杂环丁基；
    更优选地，R2为-H、-CN、甲基、叔丁基、环丙基、氧杂环丁基或氮杂环丁基，所述甲基或叔丁基任选地被一个或化合价所允许范围内的多个-F取代，所述环丙基、氧杂环丁基或氮杂环丁基任选地被一个或者化合价所允许范围内的多个甲基或-F所取代；
    更进一步优选地，R2为-H、-CN、-CH3、-CHF2、-CF3、-CH(CH3)3、环丙基、
  11. 根据权利要求1-10任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，R3为-C1-3烷基，优选为甲基；和/或
    R4各自独立地为-F、-Cl或-Br，优选为-Cl。
  12. 根据权利要求1-11任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，所述的化合物具有以下结构
    其中，Y、L1、R2、E1～E3、X1～X3如权利要求1所述；
    优选地，Y为-CH2-，X1为CH，X2为N，X3为N。
  13. 根据权利要求1所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，所述的化合物具有以下结构
    其中，R3为-C1-3烷基；
    R5为任选被一个或多个取代基取代的5-6元杂芳基；所述取代基各自独立地选自氢、卤素、-CN、羟基、-C1-3烷基或-C1-3卤代烷基，所述5-6元杂芳基含1、2或3个杂原子。
  14. 根据权利要求1-13任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，所述的化合物选自以下结构式所示的化合物：



  15. 根据权利要求1-13任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，其特征在于，所述的化合物选自以下结构式所示的化合物：


  16. 根据权利要求1-15任一项所述的化合物、其立体异构体、互变异构体或其可药用盐的制备方法，其特征在于，包含以下方案一～方案三中任一方案所示步骤：
    方案一：
    方案二：当X3为CRa时
    方案三：当X3为N时
    其中，各基团如权利要求1所述。
  17. 一种药物组合物，其特征在于，包括根据权利要求1-15任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，以及任选一种或多种药学上可接受的载体和/或赋形剂。
  18. 根据权利要求1-15任一项所述的化合物、其立体异构体、互变异构体或其可药用盐或根据权利要求17所述的药物组合物在制备用于预防和/或治疗CCR4介导的疾病的药物中的用途；优选地，所述CCR4介导的疾病包括炎性疾病、自身免疫性疾病或癌症；更优选地，所述炎性疾病和自身免疫疾病为皮炎、过敏性哮喘、炎症性肠病或红斑狼疮，所述癌症为肝癌、胰腺癌、结肠癌、肺癌、脑癌或胃癌。
  19. 一种治疗CCR4介导的疾病的方法，其特征在于，包括给需要这种治疗的患者或有需要的个体施用治疗有效量的任一据权利要求1-15任一项所述的化合物、其立体异构体、互变异构体或其可药用盐，或者根据权利要求17所述的药物组合物。
PCT/CN2024/130449 2023-11-09 2024-11-07 趋化因子受体调节剂制备方法和用途 Pending WO2025098425A1 (zh)

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