WO2023016527A1 - Class of benzoxazine spiro compounds and preparation method therefor - Google Patents

Class of benzoxazine spiro compounds and preparation method therefor Download PDF

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Publication number
WO2023016527A1
WO2023016527A1 PCT/CN2022/111883 CN2022111883W WO2023016527A1 WO 2023016527 A1 WO2023016527 A1 WO 2023016527A1 CN 2022111883 W CN2022111883 W CN 2022111883W WO 2023016527 A1 WO2023016527 A1 WO 2023016527A1
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compound
reaction solution
alkyl
added
mmol
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PCT/CN2022/111883
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French (fr)
Chinese (zh)
Inventor
吴凌云
赵乐乐
孙建军
贾海飞
黎健
陈曙辉
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南京明德新药研发有限公司
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Publication of WO2023016527A1 publication Critical patent/WO2023016527A1/en

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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
    • A61K31/397—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having four-membered rings, e.g. azetidine
    • 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
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/538—1,4-Oxazines, e.g. morpholine ortho- or peri-condensed with carbocyclic ring systems
    • 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
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5386—1,4-Oxazines, e.g. morpholine spiro-condensed or forming part of bridged ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/10—Spiro-condensed systems

Definitions

  • the invention relates to a class of benzoxazine spiro compounds, a preparation method thereof, and the application of the compounds in the preparation of medicines for treating related diseases. It specifically relates to the compound represented by formula (I) and its pharmaceutically acceptable salt.
  • Tumor immunotherapy is a therapeutic area that has attracted much attention in recent years.
  • the main mechanism is to enhance the anti-tumor ability of the immune microenvironment by mobilizing the body's immune system.
  • monoclonal antibody drugs for tumor immunotherapy such as Keytruda and OPDIVO, have been used for the treatment of various cancers such as non-small cell lung cancer and melanoma.
  • RORs Retinoic acid-related orphan receptors
  • RORs Retinoic acid-related orphan receptors
  • RORs belong to the nuclear receptor superfamily and are a member of intracellular transcription factors, which can regulate a variety of physiological processes, including reproductive development, metabolism, and immune system regulation.
  • ROR has three family members: ROR- ⁇ , - ⁇ and - ⁇ , which are encoded by RORA, RORB and RORC genes, respectively.
  • ROR ⁇ includes two subtypes, ROR ⁇ 1 and ROR ⁇ t (ROR ⁇ 2).
  • ROR ⁇ 1 is expressed in various tissues and organs such as thymus, muscle, pancreas, prostate, and liver, while the short-chain subtype ROR ⁇ t of ROR ⁇ is mainly distributed in the thymus and promotes the differentiation of initial T cells into Th17 and Tc17 cells.
  • Th17 and Tc17 cells promote inflammatory and autoimmune responses by secreting IL-17, IL-22, GM-CSF and other cytokines and inflammatory factors.
  • IL-17 can promote the recruitment and infiltration of CTLs and NK cells in the tumor microenvironment, and enhance the anti-tumor effect of effector T cells.
  • ROR ⁇ agonists currently have no drugs approved for marketing, and Lycera Corp’s
  • ROR ⁇ agonist LYC-55716 monotherapy for the treatment of advanced solid tumors is in phase II clinical research, and it is combined with PD-1 monoclonal antibody pembrolizumab in the treatment of advanced non-small cell lung cancer The treatment is in a phase 1b clinical study.
  • this field still needs candidate compounds with better activity and better pharmacokinetic parameters to advance to clinical trials to meet the therapeutic needs.
  • the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,
  • R 11 , R 12 , R 13 , R 14 and R 15 are independently selected from H, F, Cl, Br, I, -OH, -NH 2 , -CN, C 1-3 alkyl, C 1-3 Alkoxy and C 1-3 alkylamino, wherein said C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino are independently optionally substituted by 1, 2 or 3 R a ;
  • R 2 is selected from C 1-6 alkyl and -C 1-3 alkyl-C 3-4 cycloalkyl, said C 1-6 alkyl and -C 1-3 alkyl-C 3-4 cycloalkane
  • the groups are independently optionally substituted by 1, 2 or 3 R b ;
  • L is selected from a single bond, -C 2-4 alkenyl- and -4-5 membered heterocycloalkyl-, and the -C 2-4 alkenyl- and -4-5 membered heterocycloalkyl- are independently optionally substituted by 1, 2 or 3 R c ;
  • R a is independently selected from F, Cl, Br, I, -OH, -NH 2 and C 1-3 alkyl, wherein the C 1-3 alkyl is optionally substituted by 1, 2 or 3 R;
  • R b are independently selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and C 1-3 alkyl;
  • R c are independently selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and C 1-3 alkyl;
  • R is independently selected from F, Cl, Br, I, -OH and -NH 2 ;
  • Hetero in the -4-5 membered heterocycloalkyl- means 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from -O-, -NH-, -S- and N, respectively.
  • R a is selected from F, Cl, Br, I, -OH, -NH 2 and -CH 3 , and other variables are as defined in the present invention.
  • R a is selected from F, and other variables are as defined in the present invention.
  • R b is selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and -CH 3 , and other variables are as defined in the present invention.
  • R b is selected from -COOH, and other variables are as defined in the present invention.
  • R b is selected from -CH 3 , and other variables are as defined in the present invention.
  • R c is selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and -CH 3 , and other variables are as defined in the present invention.
  • R c is selected from -CH 3 , and other variables are as defined in the present invention.
  • R 11 , R 12 , R 13 , R 14 and R 15 are independently selected from H, F, Cl, Br, I, -OH, -NH 2 , -CN, -CH 3 , -O-CH 3 and -NH-CH 3 , wherein said -CH 3 , -O-CH 3 and -NH-CH 3 are independently optionally substituted by 1, 2 or 3 R a , R a and Other variables are as defined herein.
  • R 11 , R 12 , R 13 , R 14 and R 15 are selected from H, F, Cl, Br, I, -OH, -NH 2 , -CN, -CH 3 , -CF 3 , -O-CH 3 , -O-CH 2 F, -O-CHF 2 , -O-CF 3 and Other variables are as defined herein.
  • R 11 is selected from H, Cl and -O-CH 3 , and other variables are as defined in the present invention.
  • R 12 is selected from H and F, and other variables are as defined in the present invention.
  • R 13 is selected from H, and other variables are as defined in the present invention.
  • R 14 is selected from H and -O-CH 3 , wherein the -O-CH 3 is optionally substituted by 1, 2 or 3 R a , and R a and other variables are as described in the present invention definition.
  • R 14 is selected from H and -O-CHF 2 , and other variables are as defined in the present invention.
  • the above-mentioned R 15 is selected from H, F and -CH 3 , wherein the -CH 3 is optionally substituted by 1, 2 or 3 R a , and R a and other variables are as defined in the present invention.
  • R 15 is selected from H, F and -CF 3 , and other variables are as defined in the present invention.
  • R 2 is selected from C 1-4 alkyl and -C 1-3 alkyl-cyclopropyl, wherein the C 1-4 alkyl and -C 1-3 alkyl-cyclopropyl
  • the propyl groups are each independently optionally substituted by 1, 2 or 3 R b , and R b and other variables are as defined in the present invention.
  • R 2 is selected from -CH 3 , -CH 2 -CH 3 , -CH 2 -CH 2 -CH 3 , Wherein -CH 3 , -CH 2 -CH 3 , -CH 2 -CH 2 -CH 3 , are independently and optionally substituted by 1, 2 or 3 R b , and R b and other variables are as defined in the present invention.
  • the above-mentioned L is selected from single bonds, -C 2-3 alkenyl- and -azetidinyl-, wherein the -C 2-3 alkenyl- and -azetidinyl-are independently optionally Substituted by 1, 2 or 3 Rc , Rc and other variables are as defined herein.
  • the present invention also provides the following compounds or pharmaceutically acceptable salts thereof,
  • the compound of the present invention As a class of benzoxazine spirocyclic compounds with ROR ⁇ agonistic activity, the compound of the present invention has significant in vitro activity and good pharmacokinetic properties, and can be combined with PD-1 alone in the MC38 mouse colon cancer xenograft model. Combination of antibiotics has excellent anti-tumor effect.
  • pharmaceutically acceptable refers to those compounds, materials, compositions and/or dosage forms, which are suitable for use in contact with human and animal tissues within the scope of sound medical judgment , without undue toxicity, irritation, allergic reaction or other problems or complications, commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable salt refers to a salt of a compound of the present invention, which is prepared from a compound having a specific substituent found in the present invention and a relatively non-toxic acid or base.
  • base addition salts can be obtained by contacting such compounds with a sufficient amount of base, either neat solution or in a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts.
  • acid addition salts can be obtained by contacting such compounds with a sufficient amount of the acid, either neat solution or in a suitable inert solvent.
  • Examples of pharmaceutically acceptable acid addition salts include salts of inorganic acids including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogenphosphate, dihydrogenphosphate, sulfuric acid, Hydrogen sulfate, hydriodic acid, phosphorous acid, etc.; and organic acid salts, 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 and methanesulfonic acid and similar acids; also salts of amino acids (such as arginine, etc.) , and salts of organic acids such as glucuronic acid.
  • Certain specific compounds of the present invention contain basic and acidic functional groups and can
  • the pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing acid groups or bases by conventional chemical methods.
  • such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.
  • the compounds of the invention may exist in particular geometric or stereoisomeric forms.
  • the present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, and their racemic and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which are subject to the present within the scope of the invention.
  • Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.
  • enantiomer or “optical isomer” refer to stereoisomers that are mirror images of each other.
  • cis-trans isomers or “geometric isomers” arise from the inability to rotate freely due to the double bond or the single bond of the carbon atoms forming the ring.
  • diastereoisomer refers to stereoisomers whose molecules have two or more chiral centers and which are not mirror images of the molecules.
  • keys with wedge-shaped solid lines and dotted wedge keys Indicates the absolute configuration of a stereocenter, with a straight solid-line bond and straight dashed keys Indicates the relative configuration of the stereocenter, with a wavy line Indicates wedge-shaped solid-line bond or dotted wedge key or with tilde Indicates a straight solid line key and straight dashed keys
  • tautomer or “tautomeric form” means that isomers with different functional groups are in dynamic equilibrium at room temperature and are rapidly interconvertible. If tautomerism is possible (eg, in solution), then chemical equilibrium of the tautomers can be achieved.
  • proton tautomers also called prototropic tautomers
  • prototropic tautomers include interconversions via migration of a proton, such as keto-enol isomerization and imine-ene Amine isomerization.
  • Valence isomers (valence tautomers) involve interconversions by recombination of some bonding electrons.
  • keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.
  • the terms “enriched in an isomer”, “enriched in an isomer”, “enriched in an enantiomer” or “enantiomerically enriched” refer to one of the isomers or enantiomers
  • the content of the enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or Greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.
  • the terms “isomer excess” or “enantiomeric excess” refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomeric excess (ee value) is 80% .
  • Optically active (R)- and (S)-isomers as well as D and L-isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the invention is desired, it can be prepared by asymmetric synthesis or derivatization with chiral auxiliary agents, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide pure desired enantiomer.
  • a diastereoisomeric salt is formed with an appropriate optically active acid or base, and then a diastereomeric salt is formed by a conventional method known in the art. Diastereomeric resolution is performed and the pure enantiomers are recovered. Furthermore, the separation of enantiomers and diastereomers is usually accomplished by the use of chromatography using chiral stationary phases, optionally in combination with chemical derivatization methods (e.g. from amines to amino groups formate).
  • the compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compounds.
  • compounds may be labeled with radioactive isotopes such as tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C).
  • radioactive isotopes such as tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C).
  • heavy hydrogen can be used to replace hydrogen to form deuterated drugs.
  • the bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon.
  • deuterated drugs can reduce toxic side effects and increase drug stability. , enhance the efficacy, prolong the biological half-life of drugs and other advantages. All changes in isotopic composition of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.
  • substituted means that any one or more hydrogen atoms on a specified atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence of the specified atom is normal and the substituted compound is stable.
  • any variable eg, R
  • its definition is independent at each occurrence.
  • said group may optionally be substituted with up to two R, with independent options for each occurrence of R.
  • substituents and/or variations thereof are permissible only if such combinations result in stable compounds.
  • linking group When the number of a linking group is 0, such as -(CRR) 0 -, it means that the linking group is a single bond.
  • substituent When a substituent is vacant, it means that the substituent does not exist. For example, when X in A-X is vacant, it means that the structure is actually A. When the enumerated substituent does not indicate which atom it is connected to the substituted group, this substituent can be bonded through any atom, for example, pyridyl as a substituent can be connected to any atom on the pyridine ring. The carbon atom is attached to the group being substituted.
  • linking group listed does not indicate its linking direction
  • its linking direction is arbitrary, for example,
  • the connecting group L in the middle is -MW-, at this time -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form It can also be formed by connecting loop A and loop B in the opposite direction to the reading order from left to right
  • any one or more sites of the group can be linked to other groups through chemical bonds.
  • connection method of the chemical bond is not positioned, and there is an H atom at the connectable site, when the chemical bond is connected, the number of H atoms at the site will decrease correspondingly with the number of chemical bonds connected to become the corresponding valence group.
  • the chemical bonds that the site connects with other groups can use straight solid line bonds Straight dotted key or tilde express.
  • the straight-shaped solid-line bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group;
  • the straight dotted line bond indicates that the two ends of the nitrogen atom in the group are connected to other groups;
  • the wavy lines in indicate that the 1 and 2 carbon atoms in the phenyl group are connected to other groups;
  • the number of atoms in a ring is generally defined as the number of ring members, eg, "5-7 membered ring” means a “ring” with 5-7 atoms arranged around it.
  • C 1-3 alkyl is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms.
  • the C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine) .
  • Examples of C 1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n - propyl and isopropyl), and the like.
  • C 1-4 alkyl is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 4 carbon atoms.
  • the C 1-4 alkyl group includes C 1-2 , C 1-3 and C 2-3 alkyl groups, etc.; it may be monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine).
  • Examples of C 1-4 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , s-butyl and t-butyl) and so on.
  • C 1-6 alkyl is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms.
  • the C 1-6 alkyl group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 and C 5 alkyl, etc.; it can be Is monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine).
  • C 1-6 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, etc.
  • C2-4alkenyl is used to denote a straight or branched chain hydrocarbon group consisting of 2 to 4 carbon atoms containing at least one carbon-carbon double bond, a carbon-carbon double bond can be located anywhere in the group.
  • the C 2-4 alkenyl includes C 2-3 , C 4 , C 3 and C 2 alkenyl, etc.; the C 2-4 alkenyl can be monovalent, divalent or multivalent. Examples of C alkenyl include, but are not limited to, ethenyl, propenyl, butenyl, butadienyl, and the like.
  • C2-3 alkenyl is used to denote a straight or branched chain hydrocarbon group consisting of 2 to 3 carbon atoms containing at least one carbon-carbon double bond, a carbon-carbon double bond It can be located at any position of the group, and the C 2-3 alkenyl includes C 3 and C 2 alkenyl; the C 2-3 alkenyl can be monovalent, divalent or multivalent. Examples of C 2-3 alkenyl include, but are not limited to, ethenyl, propenyl, and the like.
  • C 1-3 alkoxy denotes those alkyl groups containing 1 to 3 carbon atoms attached to the rest of the molecule through an oxygen atom.
  • the C 1-3 alkoxy group includes C 1-2 , C 2-3 , C 3 and C 2 alkoxy groups and the like.
  • Examples of C 1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.
  • C 1-3 alkylamino denotes those alkyl groups containing 1 to 3 carbon atoms attached to the rest of the molecule through an amino group.
  • the C 1-3 alkylamino group includes C 1-2 , C 3 and C 2 alkylamino groups and the like.
  • Examples of C 1-3 alkylamino include, but are not limited to, -NHCH 3 , -N(CH 3 ) 2 , -NHCH 2 CH 3 , -N(CH 3 )CH 2 CH 3 , -NHCH 2 CH 2 CH 3 , -NHCH 2 CH 2 CH 3 , -NHCH 2 (CH 3 ) 2 etc.
  • C 3-4 cycloalkyl means a saturated cyclic hydrocarbon group composed of 3 to 4 carbon atoms, which is a monocyclic ring system; it may be monovalent, divalent or multivalent.
  • Examples of C 3-4 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl and the like.
  • a heteroatom may occupy the attachment position of the heterocycloalkyl to the rest of the molecule.
  • the 4-5 membered heterocycloalkyl group includes 4-membered and 5-membered heterocycloalkyl groups.
  • Examples of 4-5 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.) or tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.) and the like.
  • C n-n+m or C n -C n+m includes any specific instance of n to n+m carbons, for example C 1-12 includes C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 , and C 12 , also including any range from n to n+m, for example, C 1-12 includes C 1- 3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12 etc.; similarly, n to n +m means that the number of atoms on the ring is n to n+m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membere
  • leaving group refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (eg, a nucleophilic substitution reaction).
  • a substitution reaction eg, a nucleophilic substitution reaction
  • representative leaving groups include triflate; chlorine, bromine, iodine; sulfonate groups such as mesylate, tosylate, brosylate, tosylate esters, etc.; acyloxy groups such as acetoxy, trifluoroacetoxy, and the like.
  • protecting group includes, but is not limited to, "amino protecting group", “hydroxyl protecting group” or “mercapto protecting group”.
  • amino protecting group refers to a protecting group suitable for preventing side reactions at the amino nitrogen position.
  • Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (such as acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, such as tert-butoxycarbonyl (Boc) ; arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethyloxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-di -(4'-methoxyphenyl)methyl; silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS) and the like.
  • acyl such as alkanoyl (such as acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, such as
  • hydroxyl protecting group refers to a protecting group suitable for preventing side reactions of the hydroxy group.
  • Representative hydroxy protecting groups include, but are not limited to: alkyl groups such as methyl, ethyl, and tert-butyl; acyl groups such as alkanoyl (such as acetyl); arylmethyl groups such as benzyl (Bn), p-formyl Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyl Dimethylsilyl (TBS) and the like.
  • alkyl groups such as methyl, ethyl, and tert-butyl
  • acyl groups such as alkanoyl (such as acetyl)
  • arylmethyl groups such as benzyl (Bn), p-formyl Oxybenzyl (P
  • the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and the methods well known to those skilled in the art Equivalent alternatives, preferred embodiments include but are not limited to the examples of the present invention.
  • the structure of the compounds of the present invention can be confirmed by conventional methods known to those skilled in the art. If the present invention involves the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, in single crystal X-ray diffraction (SXRD), the cultured single crystal is collected with a Bruker D8 venture diffractometer to collect diffraction intensity data, the light source is CuK ⁇ radiation, and the scanning method is: After scanning and collecting relevant data, the absolute configuration can be confirmed by further analyzing the crystal structure by direct method (Shelxs97).
  • SXRD single crystal X-ray diffraction
  • aq stands for water
  • HATU O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphonic acid Salt
  • EDC stands for N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride
  • m-CPBA stands for 3-chloroperoxybenzoic acid
  • eq stands for equivalent, equivalent
  • CDI DCM stands for dichloromethane
  • PE stands for petroleum ether
  • DIAD diisopropyl azodicarboxylate
  • DMF stands for N,N-dimethylformamide
  • DMSO stands for dimethylsulfoxide
  • EtOAc stands for acetic acid EtOH stands for ethanol
  • MeOH stands for methanol
  • CBz stands for benzyloxycarbonyl, an amine protecting group
  • compound 1-1 (2.20g, 10.6mmol) was dissolved in anhydrous methanol (20mL), potassium carbonate (2.92g, 21.1mmol) and 1-2 (2.43g, 12.7mmol) were added, and the reaction solution After stirring at 25°C for 12 hours, water (40 mL) was added to the reaction solution, and extracted with ethyl acetate (50 mL x 1). The organic phase was washed with saturated brine (50mL x 1), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (50:1 petroleum ether/ethyl acetate) to obtain compound 1-3 .
  • 1 H NMR 400 MHz, CDCl 3
  • ⁇ 7.56-7.51 (m, 2H), 7.33-7.29 (m, 1H), 3.64 (s, 1H).
  • compound 1-3 (1.38g, 6.75mmol) was dissolved in anhydrous methanol (15mL), then 1-4 (2.57g, 10.1mmol), copper powder (42.9mg, 0.675mmol) and potassium carbonate were added (1.86g, 13.5mmol), the reaction solution was stirred at 25°C for 12 hours, the reaction solution was spin-dried, added ethyl acetate (50mL), filtered, the filter cake was washed with ethyl acetate (10mL), and the filtrate was concentrated under reduced pressure , the residue was separated and purified by silica gel column chromatography (20:1 petroleum ether/ethyl acetate) to obtain compound 1-5.
  • diboronic acid pinacol ester (819mg, 3.23mmol) was added into anhydrous tetrahydrofuran (10mL), then cuprous chloride (29.0mg, 0.293mmol) and 4,5-bis(diphenylphosphine) were added -9,9-Dimethylxanthene (170mg, 0.293mmol), the reaction solution was stirred at 25°C for 5 minutes, then sodium tert-butoxide (310mg, 3.23mmol) was added, and the reaction solution was stirred at 25°C for 5 minutes , then added compound 1-3 (600mg, 2.93mmol) and iodomethane (1.67g, 11.7mmol), the reaction solution was stirred at 25°C for 710 minutes, water (20mL) was added to the reaction solution, and ethyl acetate (20mL x 1) extraction, the organic phase was washed with saturated brine (20mLx 1), dried over anhydrous sodium sulf
  • Zinc dust (1.81 g, 27.7 mmol) and 1,2-dibromoethane (307 mg, 1.63 mmol) were added to N,N-dimethylformamide (20 mL).
  • the reaction solution was stirred and reacted at 70° C. for 10 minutes.
  • the reaction solution was then cooled to 20°C.
  • Chlorotrimethylsilane (177mg, 1.63mmol) was added dropwise to the reaction solution, and then the reaction solution was stirred at 20°C for 50 minutes.
  • reaction solution was filtered through diatomaceous earth, the filter residue was washed with ethyl acetate (30mL), the filtrate was washed with saturated ammonium chloride aqueous solution (30mL ⁇ 1), and the organic phase was sequentially washed with water (30mL ⁇ 3) and saturated brine (30mL ⁇ 1). After washing, drying with anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (4/1 petroleum ether/ethyl acetate) to obtain compound 15-3.
  • ROR ⁇ ligand-binding domain LBD
  • TR-FRET time-resolved fluorescence energy transfer
  • the compound to be tested was diluted in DMSO and further diluted in assay buffer (50mM Tris pH 7.0, 50mM KCl, 1mM Na-EDTA, 0.1mM DTT, 0.01% BSA) (4-fold dilution, 10 concentrations, highest concentration 5000 nM), the final DMSO concentration was 1%.
  • the hROR ⁇ -LBD protein was diluted in assay buffer to give a final hROR ⁇ -LBD concentration of 15 nM in 384-well plates.
  • a stock solution of biotin-SRC1 polypeptide Biotin-SPSSHSSSLTERHKILHRLLQEGSP was prepared in assay buffer and added to each well (final concentration 200 nM). Solutions of SA-eu (1 nM final concentration) and SA-APC (50 nM final concentration) were also added to each well.
  • the final assay mixture was incubated overnight at 4°C, equilibrated at room temperature for 1 hour, and centrifuged at 1000 rpm for 1 minute. Fluorescence readings were detected on the Envision microplate detector, and the logarithmic curve of the ratio of the fluorescence signal at emission wavelength 665nM/615nM to the compound concentration was drawn by GraphPad Prism software, and the 50% effective concentration (EC 50 ) of the compound was calculated. The maximum response (E max ) was the upper peak peak of the signal determined by GraphPad Prism fit.
  • Mouse CD4+T cell isolation kit (Mouse CD4+T cell isolation kit) (Stemcell)
  • Non-essential amino acids (Gibco)
  • the CD3 antibody was diluted to 5 ⁇ g/mL in DPBS, added to a 96-well U-bottom plate, 50 ⁇ L of liquid per well, and coated overnight at 4°C.
  • C57BL/6 mouse spleen in culture medium RPMI 1640 + 10% fetal bovine serum + 1% penicillin + 1% non-essential amino acid + 0.05mM ⁇ -mercaptoethanol
  • RPMI 1640 + 10% fetal bovine serum + 1% penicillin + 1% non-essential amino acid + 0.05mM ⁇ -mercaptoethanol pass through a 70 ⁇ m filter to prepare a single cell suspension , centrifuge at 300g for 3min. Add erythrocyte lysate to lyse at room temperature for 3 min.
  • CD4 + cells were isolated using the mouse CD4 + T cell isolation kit.
  • CD4 + cells obtained above at a density of 5*10 5 /mL into the coated wells , 200 ⁇ L cell suspension per well; then add CD28 antibody (3 ⁇ g/mL), TGF ⁇ (3ng/mL), IL-6 (30ng/mL), IL-23 (10ng/mL), IL-1 ⁇ (10ng/mL ), IFN ⁇ antibody (10 ⁇ g/mL) and IL-4 antibody (10 ⁇ g/mL); then the compound of the present invention was added to the well, and cultured at 37° C. under 5% CO 2 for 3 days.
  • the U-bottom plate was centrifuged at 300 g for 3 min, the supernatant was discarded, and washed twice with staining buffer.
  • the IL-17A antibody was diluted 1:200 in permeabilization buffer, 50 ⁇ L of dye solution was added to each well, stained at room temperature for 30 min, and then washed twice with staining buffer. Finally, the cells were resuspended with 150 ⁇ L staining buffer, and the proportion of Th17 cells was detected by flow cytometry.
  • Compound test concentration 1 ⁇ M.
  • the compounds of the present invention can obviously promote the differentiation of CD4 + cells into Th17 cells.
  • the pharmacokinetic characteristics of the compounds were tested in rodents after intravenous injection and oral administration according to the standard protocol.
  • the mice were given a single intravenous injection (IV) and oral administration (PO).
  • the solvent for intravenous injection is a mixed solvent made up of 5% dimethyl sulfoxide, 30% PEG400, and 65% 10% hydroxypropyl ⁇ -cyclodextrin.
  • the oral vehicle is a mixed vehicle made of 0.5% hypromellose and 0.2% Tween.
  • the project used four female Balb/c mice, two mice were administered intravenously, the dose was 0.5mg/kg, and the collection 0h (before administration) and after administration were 0.0833, 0.25, 0.5, 1, Plasma samples at 2, 4, 8, and 24 hours were administered orally to the other two mice at a dose of 1 mg/kg, collected at 0 h (before administration) and at 0.25, 0.5, 1, 2, and 4 hours after administration , 8, 24h plasma samples, collect whole blood samples within 24 hours, centrifuge at 3000g for 15 minutes, separate supernatant to obtain plasma samples, add 4 times volume of acetonitrile solution containing internal standard to precipitate protein, centrifuge to take supernatant and add equal volume The water was then centrifuged to take the supernatant sample, and the blood drug concentration was quantitatively analyzed by LC-MS/MS analysis method, and the pharmacokinetic parameters were calculated, such as peak concentration (C max ), clearance rate (CL), half-life (T 1 / 2 ), tissue distribution (V
  • the compounds of this invention have good pharmacokinetic properties, including good oral bioavailability, oral exposure, half-life and clearance rate.
  • the purpose of this experiment is to study the evaluation of the compound of the present invention on the MC38 mouse colon cancer xenograft tumor model in vivo.
  • the culture medium is 1640 medium containing 10% fetal bovine serum, and the culture condition is 37°C, 5% carbon dioxide.
  • the subculture ratio was 1:2 ⁇ 1:3, and subcultured 2 ⁇ 3 times a week.
  • 0.1 mL (2 ⁇ 10 5 ) cells were inoculated subcutaneously on the right back of each mouse. On the same day, animals were randomized into groups based on body weight.
  • the experimental vehicle was 5% DMSO/95% (20% hydroxypropyl beta cyclodextrin).
  • the test substance was dissolved in a solvent, prepared into a uniform solution with a certain concentration, and stored at 4°C.
  • the experimental index is to investigate whether tumor growth is inhibited, delayed or cured.
  • Tumor diameters were measured twice a week with vernier calipers.
  • T/C Relative tumor proliferation rate
  • RTV relative tumor volume
  • PD-1 monoclonal antibody BioXcell. PD-1 monoclonal antibody was administered on the 7th day after grouping, and compound 1 was administered on the day of grouping.
  • the combination of the compound of the present invention and PD-1 monoclonal antibody has an excellent tumor-inhibiting effect on the transplanted tumor model of MC38 mouse colon cancer.

Abstract

Provided are a class of benzoxazine spiro compounds and a preparation method therefor, and the use of the class of compounds in the preparation of a drug for treating related diseases. Specifically, disclosed are a compound as represented by formula (I) and a pharmaceutically acceptable salt thereof.

Description

一类苯并噁嗪螺环类化合物及其制备方法A kind of benzoxazine spiro compound and preparation method thereof
本申请主张如下优先权：This application claims the following priority:
CN202110933009.8，2021年08月13日。CN202110933009.8, August 13, 2021.
技术领域technical field
本发明涉及一类苯并噁嗪螺环类化合物及其制备方法，以及该类化合物在制备治疗相关疾病药物中的应用。具体涉及式(Ⅰ)所示化合物及其药学上可接受的盐。The invention relates to a class of benzoxazine spiro compounds, a preparation method thereof, and the application of the compounds in the preparation of medicines for treating related diseases. It specifically relates to the compound represented by formula (I) and its pharmaceutically acceptable salt.
背景技术Background technique
肿瘤免疫疗法是近年来备受关注的治疗领域，主要机理是通过调动机体的免疫系统，增强免疫微环境的抗肿瘤能力。目前已有肿瘤免疫治疗的单抗药物如Keytruda和OPDIVO等用于非小细胞肺癌和黑色素瘤等多种癌症的治疗。Tumor immunotherapy is a therapeutic area that has attracted much attention in recent years. The main mechanism is to enhance the anti-tumor ability of the immune microenvironment by mobilizing the body's immune system. Currently, monoclonal antibody drugs for tumor immunotherapy, such as Keytruda and OPDIVO, have been used for the treatment of various cancers such as non-small cell lung cancer and melanoma.
维甲酸相关孤儿受体(RAR-related orphan receptor，RORs)属于核受体超家族，是细胞内转录因子的一员，能够调控多种生理过程，包括生殖发育、新陈代谢、免疫系统调节等。ROR有三个家族成员：ROR-α、-β和-γ，分别由RORA、RORB、RORC基因编码。RORγ又包括RORγ1和RORγt(RORγ2)两种亚型。RORγ1在胸腺、肌肉、胰腺、前列腺和肝脏等多种组织和器官中表达，而RORγ的短链亚型RORγt主要分布于胸腺，促进初始T细胞向Th17和Tc17细胞分化。Th17和Tc17细胞通过分泌IL-17，IL-22，GM-CSF等细胞因子和炎症因子，促进炎症反应和自身免疫应答。IL-17可以促进肿瘤微环境中CTLs和NK细胞的招募和浸润，提升效应T细胞的抗肿瘤作用。Retinoic acid-related orphan receptors (RAR-related orphan receptors, RORs) belong to the nuclear receptor superfamily and are a member of intracellular transcription factors, which can regulate a variety of physiological processes, including reproductive development, metabolism, and immune system regulation. ROR has three family members: ROR-α, -β and -γ, which are encoded by RORA, RORB and RORC genes, respectively. RORγ includes two subtypes, RORγ1 and RORγt (RORγ2). RORγ1 is expressed in various tissues and organs such as thymus, muscle, pancreas, prostate, and liver, while the short-chain subtype RORγt of RORγ is mainly distributed in the thymus and promotes the differentiation of initial T cells into Th17 and Tc17 cells. Th17 and Tc17 cells promote inflammatory and autoimmune responses by secreting IL-17, IL-22, GM-CSF and other cytokines and inflammatory factors. IL-17 can promote the recruitment and infiltration of CTLs and NK cells in the tumor microenvironment, and enhance the anti-tumor effect of effector T cells.
RORγ激动剂目前没有药物获批上市，Lycera Corp公司的RORγ激动剂LYC-55716单药用于晚期实体瘤的治疗处于临床二期研究，与PD-1单抗pembrolizumab联用治疗晚期非小细胞肺癌的治疗处于临床1b期研究。面对巨大的未满足市场，该领域仍然需要活性更好，药代动力学参数更优的候选化合物推进临床试验，以满足治疗需求。RORγ agonists currently have no drugs approved for marketing, and Lycera Corp’s RORγ agonist LYC-55716 monotherapy for the treatment of advanced solid tumors is in phase II clinical research, and it is combined with PD-1 monoclonal antibody pembrolizumab in the treatment of advanced non-small cell lung cancer The treatment is in a phase 1b clinical study. In the face of a huge unmet market, this field still needs candidate compounds with better activity and better pharmacokinetic parameters to advance to clinical trials to meet the therapeutic needs.
发明内容Contents of the invention
本发明提供了式(Ⅰ)化合物或其药学上可接受的盐，The present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,
Figure PCTCN2022111883-appb-000001
Figure PCTCN2022111883-appb-000001
其中，in,
R 11、R 12、R 13、R 14和R 15分别独立地选自H、F、Cl、Br、I、-OH、-NH 2、-CN、C 1-3烷基、C 1-3烷氧基和C 1-3烷氨基，其中所述C 1-3烷基、C 1-3烷氧基和C 1-3烷氨基分别独立地任选被1、2或3个R a取代； R 11 , R 12 , R 13 , R 14 and R 15 are independently selected from H, F, Cl, Br, I, -OH, -NH 2 , -CN, C 1-3 alkyl, C 1-3 Alkoxy and C 1-3 alkylamino, wherein said C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino are independently optionally substituted by 1, 2 or 3 R a ;
R 2选自C 1-6烷基和‐C 1-3烷基-C 3-4环烷基，所述C 1-6烷基和‐C 1-3烷基-C 3-4环烷基分别独立地任选被1、2或3个R b取代； R 2 is selected from C 1-6 alkyl and -C 1-3 alkyl-C 3-4 cycloalkyl, said C 1-6 alkyl and -C 1-3 alkyl-C 3-4 cycloalkane The groups are independently optionally substituted by 1, 2 or 3 R b ;
L 1选自单键、-C 2-4烯基-和-4-5元杂环烷基-，所述-C 2-4烯基-和-4-5元杂环烷基-分别独立地任选被1、2或3个R c取代； L is selected from a single bond, -C 2-4 alkenyl- and -4-5 membered heterocycloalkyl-, and the -C 2-4 alkenyl- and -4-5 membered heterocycloalkyl- are independently optionally substituted by 1, 2 or 3 R c ;
R a分别独立地选自F、Cl、Br、I、-OH、-NH 2和C 1-3烷基，其中所述C 1-3烷基任选被1、2或3个R取代； R a is independently selected from F, Cl, Br, I, -OH, -NH 2 and C 1-3 alkyl, wherein the C 1-3 alkyl is optionally substituted by 1, 2 or 3 R;
R b分别独立地选自F、Cl、Br、I、-OH、-NH 2、-CN、-COOH和C 1-3烷基； R b are independently selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and C 1-3 alkyl;
R c分别独立地选自F、Cl、Br、I、-OH、-NH 2、-CN、-COOH和C 1-3烷基； R c are independently selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and C 1-3 alkyl;
R分别独立地选自F、Cl、Br、I、-OH和-NH 2； R is independently selected from F, Cl, Br, I, -OH and -NH 2 ;
所述-4-5元杂环烷基-中的“杂”表示1、2、3或4个分别独立选自-O-、-NH-、-S-和N的杂原子或杂原子团。"Hetero" in the -4-5 membered heterocycloalkyl- means 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from -O-, -NH-, -S- and N, respectively.
本发明的一些方案中，上述R a选自F、Cl、Br、I、-OH、-NH 2和-CH 3，其他变量如本发明所定义。 In some solutions of the present invention, the above R a is selected from F, Cl, Br, I, -OH, -NH 2 and -CH 3 , and other variables are as defined in the present invention.
本发明的一些方案中，上述R a选自F，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R a is selected from F, and other variables are as defined in the present invention.
本发明的一些方案中，上述R b选自F、Cl、Br、I、-OH、-NH 2、-CN、-COOH和-CH 3，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R b is selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and -CH 3 , and other variables are as defined in the present invention.
本发明的一些方案中，上述R b选自-COOH，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R b is selected from -COOH, and other variables are as defined in the present invention.
本发明的一些方案中，上述R b选自-CH 3，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R b is selected from -CH 3 , and other variables are as defined in the present invention.
本发明的一些方案中，上述R c选自F、Cl、Br、I、-OH、-NH 2、-CN、-COOH和-CH 3，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R c is selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and -CH 3 , and other variables are as defined in the present invention.
本发明的一些方案中，上述R c选自-CH 3，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R c is selected from -CH 3 , and other variables are as defined in the present invention.
本发明的一些方案中，上述R 11、R 12、R 13、R 14和R 15分别独立地选自H、F、Cl、Br、I、-OH、-NH 2、-CN、-CH 3、-O-CH 3和-NH-CH 3，其中所述-CH 3、-O-CH 3和-NH-CH 3分别独立地任选被1、2或3个R a取代，R a及其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R 11 , R 12 , R 13 , R 14 and R 15 are independently selected from H, F, Cl, Br, I, -OH, -NH 2 , -CN, -CH 3 , -O-CH 3 and -NH-CH 3 , wherein said -CH 3 , -O-CH 3 and -NH-CH 3 are independently optionally substituted by 1, 2 or 3 R a , R a and Other variables are as defined herein.
本发明的一些方案中，上述R 11、R 12、R 13、R 14和R 15选自H、F、Cl、Br、I、-OH、-NH 2、-CN、-CH 3、-CF 3、-O-CH 3、-O-CH 2F、-O-CHF 2、-O-CF 3和
Figure PCTCN2022111883-appb-000002
其他变量如本发明所定义。 In some solutions of the present invention, the above R 11 , R 12 , R 13 , R 14 and R 15 are selected from H, F, Cl, Br, I, -OH, -NH 2 , -CN, -CH 3 , -CF 3 , -O-CH 3 , -O-CH 2 F, -O-CHF 2 , -O-CF 3 and
Figure PCTCN2022111883-appb-000002
Other variables are as defined herein.
本发明的一些方案中，上述R 11选自H、Cl和-O-CH 3，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R 11 is selected from H, Cl and -O-CH 3 , and other variables are as defined in the present invention.
本发明的一些方案中，上述R 12选自H和F，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R 12 is selected from H and F, and other variables are as defined in the present invention.
本发明的一些方案中，上述R 13选自H，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R 13 is selected from H, and other variables are as defined in the present invention.
本发明的一些方案中，上述R 14选自H和-O-CH 3，其中所述-O-CH 3任选被1、2或3个R a取代，R a及其他变量如本发明所定义。 In some schemes of the present invention, the above-mentioned R 14 is selected from H and -O-CH 3 , wherein the -O-CH 3 is optionally substituted by 1, 2 or 3 R a , and R a and other variables are as described in the present invention definition.
本发明的一些方案中，上述R 14选自H和-O-CHF 2，其他变量如本发明所定义。 In some solutions of the present invention, the above R 14 is selected from H and -O-CHF 2 , and other variables are as defined in the present invention.
本发明的一些方案中，上述R 15选自H、F和-CH 3，其中所述-CH 3任选被1、2或3个R a取代，R a及其他变量如本发明所定义。 In some embodiments of the present invention, the above-mentioned R 15 is selected from H, F and -CH 3 , wherein the -CH 3 is optionally substituted by 1, 2 or 3 R a , and R a and other variables are as defined in the present invention.
本发明的一些方案中，上述R 15选自H、F和-CF 3，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R 15 is selected from H, F and -CF 3 , and other variables are as defined in the present invention.
本发明的一些方案中，上述R 2选自C 1-4烷基和‐C 1-3烷基-环丙基，其中所述C 1-4烷基和‐C 1-3烷基-环丙基分别独立地任选被1、2或3个R b取代，R b及其他变量如本发明所定义。 In some schemes of the present invention, the above-mentioned R 2 is selected from C 1-4 alkyl and -C 1-3 alkyl-cyclopropyl, wherein the C 1-4 alkyl and -C 1-3 alkyl-cyclopropyl The propyl groups are each independently optionally substituted by 1, 2 or 3 R b , and R b and other variables are as defined in the present invention.
本发明的一些方案中，上述R 2选自-CH 3、-CH 2-CH 3、-CH 2-CH 2-CH 3、
Figure PCTCN2022111883-appb-000003
其中所述-CH 3、-CH 2-CH 3、-CH 2-CH 2-CH 3、
Figure PCTCN2022111883-appb-000004
分别独立地任选被1、2或3个R b取代，R b及其他变量如本发明所定义。 In some schemes of the present invention, the above-mentioned R 2 is selected from -CH 3 , -CH 2 -CH 3 , -CH 2 -CH 2 -CH 3 ,
Figure PCTCN2022111883-appb-000003
Wherein -CH 3 , -CH 2 -CH 3 , -CH 2 -CH 2 -CH 3 ,
Figure PCTCN2022111883-appb-000004
are independently and optionally substituted by 1, 2 or 3 R b , and R b and other variables are as defined in the present invention.
本发明的一些方案中，上述R 2选自
Figure PCTCN2022111883-appb-000005
Figure PCTCN2022111883-appb-000006
其他变量如本发明所定义。 In some schemes of the present invention, above-mentioned R 2 is selected from
Figure PCTCN2022111883-appb-000005
Figure PCTCN2022111883-appb-000006
Other variables are as defined herein.
本发明的一些方案中，上述L 1选自单键、-C 2-3烯基-和-吖丁啶基‐，其中所述-C 2-3烯基-和-吖丁啶基‐分别独立地任选被1、2或3个R c取代，R c及其他变量如本发明所定义。 In some schemes of the present invention, the above-mentioned L is selected from single bonds, -C 2-3 alkenyl- and -azetidinyl-, wherein the -C 2-3 alkenyl- and -azetidinyl-are independently optionally Substituted by 1, 2 or 3 Rc , Rc and other variables are as defined herein.
本发明的一些方案中，上述L 1选自单键、-CH＝CH-和
Figure PCTCN2022111883-appb-000007
其中所述-CH＝CH-和
Figure PCTCN2022111883-appb-000008
分别独立地任选被1、2或3个R c取代，R c及其他变量如本发明所定义。 In some schemes of the present invention, above-mentioned L 1 is selected from single bond, -CH=CH- and
Figure PCTCN2022111883-appb-000007
Wherein said -CH=CH- and
Figure PCTCN2022111883-appb-000008
are independently and optionally substituted by 1, 2 or 3 R c , R c and other variables are as defined in the present invention.
本发明的一些方案中，上述L 1选自单键、-CH＝CH-、-C(CH 3)＝CH-和
Figure PCTCN2022111883-appb-000009
其他变量如本发明所定义。 In some schemes of the present invention, the above-mentioned L 1 is selected from single bond, -CH=CH-, -C(CH 3 )=CH- and
Figure PCTCN2022111883-appb-000009
Other variables are as defined herein.
本发明的一些方案中，上述结构单元
Figure PCTCN2022111883-appb-000010
选自
Figure PCTCN2022111883-appb-000011
Figure PCTCN2022111883-appb-000012
其他变量如本发明所定义。 In some solutions of the present invention, the above structural units
Figure PCTCN2022111883-appb-000010
selected from
Figure PCTCN2022111883-appb-000011
Figure PCTCN2022111883-appb-000012
Other variables are as defined herein.
本发明还有一些方案是由上述各变量任意组合而来。Some schemes of the present invention are formed by any combination of the above-mentioned variables.
本发明还提供了下列所示化合物或其药学上可接受的盐，The present invention also provides the following compounds or pharmaceutically acceptable salts thereof,
Figure PCTCN2022111883-appb-000013
Figure PCTCN2022111883-appb-000013
技术效果technical effect
作为一类具有RORγ激动活性的苯并噁嗪螺环类化合物，本发明的化合物体外活性显著，具有良好的药代动力学性质，并且在MC38小鼠结肠癌移植瘤模型中与PD-1单抗联用具有优异的抑瘤效果。As a class of benzoxazine spirocyclic compounds with RORγ agonistic activity, the compound of the present invention has significant in vitro activity and good pharmacokinetic properties, and can be combined with PD-1 alone in the MC38 mouse colon cancer xenograft model. Combination of antibiotics has excellent anti-tumor effect.
定义和说明Definition and Description
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时， 意在指代其对应的商品或其活性成分。Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A specific term or phrase should not be considered indeterminate or unclear if it is not specifically defined, but should be understood according to its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding trade or its active ingredients.
这里所采用的术语“药学上可接受的”，是针对那些化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and/or dosage forms, which are suitable for use in contact with human and animal tissues within the scope of sound medical judgment , without undue toxicity, irritation, allergic reaction or other problems or complications, commensurate with a reasonable benefit/risk ratio.
术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物接触的方式获得碱加成盐。药学上可接受的碱加成盐包括钠、钾、钙、铵、有机胺或镁盐或类似的盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸与这类化合物接触的方式获得酸加成盐。药学上可接受的酸加成盐的实例包括无机酸盐，所述无机酸包括例如盐酸、氢溴酸、硝酸、碳酸，碳酸氢根，磷酸、磷酸一氢根、磷酸二氢根、硫酸、硫酸氢根、氢碘酸、亚磷酸等；以及有机酸盐，所述有机酸包括如乙酸、丙酸、异丁酸、马来酸、丙二酸、苯甲酸、琥珀酸、辛二酸、反丁烯二酸、乳酸、扁桃酸、邻苯二甲酸、苯磺酸、对甲苯磺酸、柠檬酸、酒石酸和甲磺酸等类似的酸；还包括氨基酸(如精氨酸等)的盐，以及如葡糖醛酸等有机酸的盐。本发明的某些特定的化合物含有碱性和酸性的官能团，从而可以被转换成任一碱或酸加成盐。The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention, which is prepared from a compound having a specific substituent found in the present invention and a relatively non-toxic acid or base. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base, either neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting such compounds with a sufficient amount of the acid, either neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts of inorganic acids including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogenphosphate, dihydrogenphosphate, sulfuric acid, Hydrogen sulfate, hydriodic acid, phosphorous acid, etc.; and organic acid salts, 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 and methanesulfonic acid and similar acids; also salts of amino acids (such as arginine, etc.) , and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups and can thus be converted into either base or acid addition salts.
本发明的药学上可接受的盐可由含有酸根或碱基的母体化合物通过常规化学方法合成。一般情况下，这样的盐的制备方法是：在水或有机溶剂或两者的混合物中，经由游离酸或碱形式的这些化合物与化学计量的适当的碱或酸反应来制备。The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing acid groups or bases by conventional chemical methods. In general, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.
本发明的化合物可以存在特定的几何或立体异构体形式。本发明设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些混合物都属于本发明的范围之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。The compounds of the invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, and their racemic and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which are subject to the present within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.
除非另有说明，术语“对映异构体”或者“旋光异构体”是指互为镜像关系的立体异构体。Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.
除非另有说明，术语“顺反异构体”或者“几何异构体”系由因双键或者成环碳原子单键不能自由旋转而引起。Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise from the inability to rotate freely due to the double bond or the single bond of the carbon atoms forming the ring.
除非另有说明，术语“非对映异构体”是指分子具有两个或多个手性中心，并且分子间为非镜像的关系的立体异构体。Unless otherwise indicated, the term "diastereoisomer" refers to stereoisomers whose molecules have two or more chiral centers and which are not mirror images of the molecules.
除非另有说明，“(+)”表示右旋，“(-)”表示左旋，“(±)”表示外消旋。Unless otherwise specified, "(+)" means dextrorotation, "(-)" means levorotation, and "(±)" means racemization.
除非另有说明，用楔形实线键
Figure PCTCN2022111883-appb-000014
和楔形虚线键
Figure PCTCN2022111883-appb-000015
表示一个立体中心的绝对构型，用直形实线键
Figure PCTCN2022111883-appb-000016
和直形虚线键
Figure PCTCN2022111883-appb-000017
表示立体中心的相对构型，用波浪线
Figure PCTCN2022111883-appb-000018
表示楔形实线键
Figure PCTCN2022111883-appb-000019
或 楔形虚线键
Figure PCTCN2022111883-appb-000020
或用波浪线
Figure PCTCN2022111883-appb-000021
表示直形实线键
Figure PCTCN2022111883-appb-000022
和直形虚线键
Figure PCTCN2022111883-appb-000023
Unless otherwise noted, keys with wedge-shaped solid lines
Figure PCTCN2022111883-appb-000014
and dotted wedge keys
Figure PCTCN2022111883-appb-000015
Indicates the absolute configuration of a stereocenter, with a straight solid-line bond
Figure PCTCN2022111883-appb-000016
and straight dashed keys
Figure PCTCN2022111883-appb-000017
Indicates the relative configuration of the stereocenter, with a wavy line
Figure PCTCN2022111883-appb-000018
Indicates wedge-shaped solid-line bond
Figure PCTCN2022111883-appb-000019
or dotted wedge key
Figure PCTCN2022111883-appb-000020
or with tilde
Figure PCTCN2022111883-appb-000021
Indicates a straight solid line key
Figure PCTCN2022111883-appb-000022
and straight dashed keys
Figure PCTCN2022111883-appb-000023
除非另有说明，术语“互变异构体”或“互变异构体形式”是指在室温下，不同官能团异构体处于动态平衡，并能很快的相互转化。若互变异构体是可能的(如在溶液中)，则可以达到互变异构体的化学平衡。例如，质子互变异构体(proton tautomer)(也称质子转移互变异构体(prototropic tautomer))包括通过质子迁移来进行的互相转化，如酮-烯醇异构化和亚胺-烯胺异构化。价键异构体(valence tautomer)包括一些成键电子的重组来进行的相互转化。其中酮-烯醇互变异构化的具体实例是戊烷-2,4-二酮与4-羟基戊-3-烯-2-酮两个互变异构体之间的互变。Unless otherwise stated, the term "tautomer" or "tautomeric form" means that isomers with different functional groups are in dynamic equilibrium at room temperature and are rapidly interconvertible. If tautomerism is possible (eg, in solution), then chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via migration of a proton, such as keto-enol isomerization and imine-ene Amine isomerization. Valence isomers (valence tautomers) involve interconversions by recombination of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.
除非另有说明，术语“富含一种异构体”、“异构体富集”、“富含一种对映体”或者“对映体富集”指其中一种异构体或对映体的含量小于100％，并且，该异构体或对映体的含量大于等于60％，或者大于等于70％，或者大于等于80％，或者大于等于90％，或者大于等于95％，或者大于等于96％，或者大于等于97％，或者大于等于98％，或者大于等于99％，或者大于等于99.5％，或者大于等于99.6％，或者大于等于99.7％，或者大于等于99.8％，或者大于等于99.9％。Unless otherwise stated, the terms "enriched in an isomer", "enriched in an isomer", "enriched in an enantiomer" or "enantiomerically enriched" refer to one of the isomers or enantiomers The content of the enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or Greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.
除非另有说明，术语“异构体过量”或“对映体过量”指两种异构体或两种对映体相对百分数之间的差值。例如，其中一种异构体或对映体的含量为90％，另一种异构体或对映体的含量为10％，则异构体或对映体过量(ee值)为80％。Unless otherwise stated, the terms "isomer excess" or "enantiomeric excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomeric excess (ee value) is 80% .
可以通过的手性合成或手性试剂或者其他常规技术制备光学活性的(R)-和(S)-异构体以及D和L异构体。如果想得到本发明某化合物的一种对映体，可以通过不对称合成或者具有手性助剂的衍生作用来制备，其中将所得非对映体混合物分离，并且辅助基团裂开以提供纯的所需对映异构体。或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的常规方法进行非对映异构体拆分，然后回收得到纯的对映体。此外，对映异构体和非对映异构体的分离通常是通过使用色谱法完成的，所述色谱法采用手性固定相，并任选地与化学衍生法相结合(例如由胺生成氨基甲酸盐)。Optically active (R)- and (S)-isomers as well as D and L-isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the invention is desired, it can be prepared by asymmetric synthesis or derivatization with chiral auxiliary agents, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereoisomeric salt is formed with an appropriate optically active acid or base, and then a diastereomeric salt is formed by a conventional method known in the art. Diastereomeric resolution is performed and the pure enantiomers are recovered. Furthermore, the separation of enantiomers and diastereomers is usually accomplished by the use of chromatography using chiral stationary phases, optionally in combination with chemical derivatization methods (e.g. from amines to amino groups formate).
本发明的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氚( 3H)，碘-125( 125I)或C-14( 14C)。又例如，可用重氢取代氢形成氘代药物，氘与碳构成的键比普通氢与碳构成的键更坚固，相比于未氘化药物，氘代药物有降低毒副作用、增加药物稳定性、增强疗效、延长药物生物半衰期等优势。本发明的化合物的所有同位素组成的变换，无论放射性与否，都包括在本发明的范围之内。 The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compounds. For example, compounds may be labeled with radioactive isotopes such as tritium ( 3 H), iodine-125 ( 125 I) or C-14 ( 14 C). For another example, heavy hydrogen can be used to replace hydrogen to form deuterated drugs. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs can reduce toxic side effects and increase drug stability. , enhance the efficacy, prolong the biological half-life of drugs and other advantages. All changes in isotopic composition of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.
术语“任选”或“任选地”指的是随后描述的事件或状况可能但不是必需出现的，并且该描述包括其中所述事件或状况发生的情况以及所述事件或状况不发生的情况。The term "optional" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where said event or circumstance occurs and instances where said event or circumstance does not occur .
术语“被取代的”是指特定原子上的任意一个或多个氢原子被取代基取代，取代基可以包括重氢和氢的 变体，只要特定原子的价态是正常的并且取代后的化合物是稳定的。当取代基为氧(即＝O)时，意味着两个氢原子被取代。氧取代不会发生在芳香基上。The term "substituted" means that any one or more hydrogen atoms on a specified atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence of the specified atom is normal and the substituted compound is stable. When a substituent is oxygen (ie =0), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups.
术语“任选被取代的”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的。The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the type and number of substituents may be arbitrary on a chemically realizable basis.
当任何变量(例如R)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。因此，例如，如果一个基团被0-2个R所取代，则所述基团可以任选地至多被两个R所取代，并且每种情况下的R都有独立的选项。此外，取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。When any variable (eg, R) occurs more than once in the composition or structure of a compound, its definition is independent at each occurrence. Thus, for example, if a group is substituted with 0-2 R, said group may optionally be substituted with up to two R, with independent options for each occurrence of R. Also, combinations of substituents and/or variations thereof are permissible only if such combinations result in stable compounds.
当一个连接基团的数量为0时，比如-(CRR) 0-，表示该连接基团为单键。 When the number of a linking group is 0, such as -(CRR) 0 -, it means that the linking group is a single bond.
当其中一个变量选自单键时，表示其连接的两个基团直接相连，比如A-L-Z中L代表单键时表示该结构实际上是A-Z。When one of the variables is selected from a single bond, it means that the two groups connected are directly connected. For example, when L in A-L-Z represents a single bond, it means that the structure is actually A-Z.
当一个取代基为空缺时，表示该取代基是不存在的，比如A-X中X为空缺时表示该结构实际上是A。当所列举的取代基中没有指明其通过哪一个原子连接到被取代的基团上时，这种取代基可以通过其任何原子相键合，例如，吡啶基作为取代基可以通过吡啶环上任意一个碳原子连接到被取代的基团上。When a substituent is vacant, it means that the substituent does not exist. For example, when X in A-X is vacant, it means that the structure is actually A. When the enumerated substituent does not indicate which atom it is connected to the substituted group, this substituent can be bonded through any atom, for example, pyridyl as a substituent can be connected to any atom on the pyridine ring. The carbon atom is attached to the group being substituted.
当所列举的连接基团没有指明其连接方向，其连接方向是任意的，例如，
Figure PCTCN2022111883-appb-000024
中连接基团L为-M-W-，此时-M-W-既可以按与从左往右的读取顺序相同的方向连接环A和环B构成
Figure PCTCN2022111883-appb-000025
也可以按照与从左往右的读取顺序相反的方向连接环A和环B构成
Figure PCTCN2022111883-appb-000026
所述连接基团、取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。 When the linking group listed does not indicate its linking direction, its linking direction is arbitrary, for example,
Figure PCTCN2022111883-appb-000024
The connecting group L in the middle is -MW-, at this time -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form
Figure PCTCN2022111883-appb-000025
It can also be formed by connecting loop A and loop B in the opposite direction to the reading order from left to right
Figure PCTCN2022111883-appb-000026
Combinations of the described linking groups, substituents and/or variations thereof are permissible only if such combinations result in stable compounds.
除非另有规定，当某一基团具有一个或多个可连接位点时，该基团的任意一个或多个位点可以通过化学键与其他基团相连。当该化学键的连接方式是不定位的，且可连接位点存在H原子时，则连接化学键时，该位点的H原子的个数会随所连接化学键的个数而对应减少变成相应价数的基团。所述位点与其他基团连接的化学键可以用直形实线键
Figure PCTCN2022111883-appb-000027
直形虚线键
Figure PCTCN2022111883-appb-000028
或波浪线
Figure PCTCN2022111883-appb-000029
表示。例如-OCH 3中的直形实线键表示通过该基团中的氧原子与其他基团相连；
Figure PCTCN2022111883-appb-000030
中的直形虚线键表示通过该基团中的氮原子的两端与其他基团相连；
Figure PCTCN2022111883-appb-000031
中的波浪线表示通过该苯基基团中的1和2位碳原子与其他基团相连；
Figure PCTCN2022111883-appb-000032
表示该哌啶基上的任意可连接位点可以通过1个化学键与其他基团相连，至少包括
Figure PCTCN2022111883-appb-000033
Figure PCTCN2022111883-appb-000034
这4种连接方式，即使-N-上画出了H原子，但是
Figure PCTCN2022111883-appb-000035
仍包括
Figure PCTCN2022111883-appb-000036
这种连接方式的基团，只是在连接1个化学键时，该位点的H会对应减少1个变成相应的一价哌啶基。 Unless otherwise specified, when a group has one or more linkable sites, any one or more sites of the group can be linked to other groups through chemical bonds. When the connection method of the chemical bond is not positioned, and there is an H atom at the connectable site, when the chemical bond is connected, the number of H atoms at the site will decrease correspondingly with the number of chemical bonds connected to become the corresponding valence group. The chemical bonds that the site connects with other groups can use straight solid line bonds
Figure PCTCN2022111883-appb-000027
Straight dotted key
Figure PCTCN2022111883-appb-000028
or tilde
Figure PCTCN2022111883-appb-000029
express. For example, the straight-shaped solid-line bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group;
Figure PCTCN2022111883-appb-000030
The straight dotted line bond in indicates that the two ends of the nitrogen atom in the group are connected to other groups;
Figure PCTCN2022111883-appb-000031
The wavy lines in indicate that the 1 and 2 carbon atoms in the phenyl group are connected to other groups;
Figure PCTCN2022111883-appb-000032
Indicates that any connectable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least
Figure PCTCN2022111883-appb-000033
Figure PCTCN2022111883-appb-000034
These 4 connection methods, even if the H atom is drawn on -N-, but
Figure PCTCN2022111883-appb-000035
still include
Figure PCTCN2022111883-appb-000036
For groups with this connection method, only when a chemical bond is connected, the H at this site will be reduced by one to become the corresponding monovalent piperidinyl group.
除非另有规定，环上原子的数目通常被定义为环的元数，例如，“5-7元环”是指环绕排列5-7个原子的“环”。Unless otherwise specified, the number of atoms in a ring is generally defined as the number of ring members, eg, "5-7 membered ring" means a "ring" with 5-7 atoms arranged around it.
除非另有规定，术语“C 1-3烷基”用于表示直链或支链的由1至3个碳原子组成的饱和碳氢基团。所述C 1-3烷基包括C 1-2和C 2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C 1- 3烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)等。 Unless otherwise specified, the term "C 1-3 alkyl" is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 alkyl group includes C 1-2 and C 2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine) . Examples of C 1-3 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n - propyl and isopropyl), and the like.
除非另有规定，术语“C 1-4烷基”用于表示直链或支链的由1至4个碳原子组成的饱和碳氢基团。所述C 1-4烷基包括C 1-2、C 1-3和C 2-3烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C 1-4烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)、丁基(包括n-丁基，异丁基，s-丁基和t-丁基)等。 Unless otherwise specified, the term "C 1-4 alkyl" is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 4 carbon atoms. The C 1-4 alkyl group includes C 1-2 , C 1-3 and C 2-3 alkyl groups, etc.; it may be monovalent (such as methyl), divalent (such as methylene) or multivalent ( such as methine). Examples of C 1-4 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , s-butyl and t-butyl) and so on.
除非另有规定，术语“C 1-6烷基”用于表示直链或支链的由1至6个碳原子组成的饱和碳氢基团。所述C 1-6烷基包括C 1-5、C 1-4、C 1-3、C 1-2、C 2-6、C 2-4、C 6和C 5烷基等；其可以是一价(如甲基)、二价(如亚甲基)或者多价(如次甲基)。C 1-6烷基的实例包括但不限于甲基(Me)、乙基(Et)、丙基(包括n-丙基和异丙基)、丁基(包括n-丁基，异丁基，s-丁基和t-丁基)、戊基(包括n-戊基，异戊基和新戊基)、己基等。 Unless otherwise specified, the term "C 1-6 alkyl" is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. The C 1-6 alkyl group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C 6 and C 5 alkyl, etc.; it can be Is monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine). Examples of C 1-6 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, etc.
除非另有规定，“C 2-4烯基”用于表示直链或支链的包含至少一个碳-碳双键的由2至4个碳原子组成的碳氢基团，碳-碳双键可以位于该基团的任何位置上。所述C 2-4烯基包括C 2-3、C 4、C 3和C 2烯基等；所述C 2-4烯基可以是一价、二价或者多价。C 2-4烯基的实例包括但不限于乙烯基、丙烯基、丁烯基、丁间二烯基等。 Unless otherwise specified, " C2-4alkenyl " is used to denote a straight or branched chain hydrocarbon group consisting of 2 to 4 carbon atoms containing at least one carbon-carbon double bond, a carbon-carbon double bond can be located anywhere in the group. The C 2-4 alkenyl includes C 2-3 , C 4 , C 3 and C 2 alkenyl, etc.; the C 2-4 alkenyl can be monovalent, divalent or multivalent. Examples of C alkenyl include, but are not limited to, ethenyl, propenyl, butenyl, butadienyl, and the like.
除非另有规定，“C 2-3烯基”用于表示直链或支链的包含至少一个碳-碳双键的由2至3个碳原子组成的碳氢基团，碳-碳双键可以位于该基团的任何位置上，所述C 2-3烯基包括C 3和C 2烯基；所述C 2-3烯基可以是一价、二价或者多价。C 2-3烯基的实例包括但不限于乙烯基、丙烯基等。 Unless otherwise specified, " C2-3 alkenyl" is used to denote a straight or branched chain hydrocarbon group consisting of 2 to 3 carbon atoms containing at least one carbon-carbon double bond, a carbon-carbon double bond It can be located at any position of the group, and the C 2-3 alkenyl includes C 3 and C 2 alkenyl; the C 2-3 alkenyl can be monovalent, divalent or multivalent. Examples of C 2-3 alkenyl include, but are not limited to, ethenyl, propenyl, and the like.
除非另有规定，术语“C 1-3烷氧基”表示通过一个氧原子连接到分子的其余部分的那些包含1至3个碳原子的烷基基团。所述C 1-3烷氧基包括C 1-2、C 2-3、C 3和C 2烷氧基等。C 1-3烷氧基的实例包括但不限于甲氧基、乙氧基、丙氧基(包括正丙氧基和异丙氧基)等。 Unless otherwise specified, the term "C 1-3 alkoxy" denotes those alkyl groups containing 1 to 3 carbon atoms attached to the rest of the molecule through an oxygen atom. The C 1-3 alkoxy group includes C 1-2 , C 2-3 , C 3 and C 2 alkoxy groups and the like. Examples of C 1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.
除非另有规定，术语“C 1-3烷氨基”表示通过氨基连接到分子的其余部分的那些包含1至3个碳原子的 烷基基团。所述C 1-3烷氨基包括C 1-2、C 3和C 2烷氨基等。C 1-3烷氨基的实例包括但不限于-NHCH 3、-N(CH 3) 2、-NHCH 2CH 3、-N(CH 3)CH 2CH 3、-NHCH 2CH 2CH 3、-NHCH 2(CH 3) 2等。 Unless otherwise specified, the term "C 1-3 alkylamino" denotes those alkyl groups containing 1 to 3 carbon atoms attached to the rest of the molecule through an amino group. The C 1-3 alkylamino group includes C 1-2 , C 3 and C 2 alkylamino groups and the like. Examples of C 1-3 alkylamino include, but are not limited to, -NHCH 3 , -N(CH 3 ) 2 , -NHCH 2 CH 3 , -N(CH 3 )CH 2 CH 3 , -NHCH 2 CH 2 CH 3 , -NHCH 2 CH 2 CH 3 , - NHCH 2 (CH 3 ) 2 etc.
除非另有规定，“C 3-4环烷基”表示由3至4个碳原子组成的饱和环状碳氢基团，其为单环体系；其可以是一价、二价或者多价。C 3-4环烷基的实例包括，但不限于，环丙基、环丁基等。 Unless otherwise specified, "C 3-4 cycloalkyl" means a saturated cyclic hydrocarbon group composed of 3 to 4 carbon atoms, which is a monocyclic ring system; it may be monovalent, divalent or multivalent. Examples of C 3-4 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl and the like.
除非另有规定，术语“4-5元杂环烷基”本身或者与其他术语联合分别表示由4至5个环原子组成的饱和单环基团，其1、2、3或4个环原子为独立选自O、S和N的杂原子，其余为碳原子，其中氮原子任选地被季铵化，碳、氮和硫杂原子可任选被氧化(即C(＝O)、NO和S(O) p，p是1或2)；其所述“4-5元杂环烷基”可以是一价、二价或者多价。此外，就该“4-5元杂环烷基”而言，杂原子可以占据杂环烷基与分子其余部分的连接位置。所述4-5元杂环烷基包括4元和5元杂环烷基。4-5元杂环烷基的实例包括但不限于氮杂环丁基、氧杂环丁基、硫杂环丁基、吡咯烷基、吡唑烷基、咪唑烷基、四氢噻吩基(包括四氢噻吩-2-基和四氢噻吩-3-基等)或四氢呋喃基(包括四氢呋喃-2-基等)等。 Unless otherwise specified, the term "4-5 membered heterocycloalkyl" by itself or in combination with other terms means a saturated monocyclic group consisting of 4 to 5 ring atoms, respectively, whose 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the carbon, nitrogen, and sulfur heteroatoms may be optionally oxidized (i.e., C(=O), NO and S(O) p , p is 1 or 2); the "4-5 membered heterocycloalkyl" can be monovalent, divalent or multivalent. In addition, with respect to the "4-5 membered heterocycloalkyl", a heteroatom may occupy the attachment position of the heterocycloalkyl to the rest of the molecule. The 4-5 membered heterocycloalkyl group includes 4-membered and 5-membered heterocycloalkyl groups. Examples of 4-5 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl ( including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.) or tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.) and the like.
除非另有规定，C n-n+m或C n-C n+m包括n至n+m个碳的任何一种具体情况，例如C 1-12包括C 1、C 2、C 3、C 4、C 5、C 6、C 7、C 8、C 9、C 10、C 11、和C 12，也包括n至n+m中的任何一个范围，例如C 1-12包括C 1- 3、C 1-6、C 1-9、C 3-6、C 3-9、C 3-12、C 6-9、C 6-12、和C 9-12等；同理，n元至n+m元表示环上原子数为n至n+m个，例如3-12元环包括3元环、4元环、5元环、6元环、7元环、8元环、9元环、10元环、11元环、和12元环，也包括n至n+m中的任何一个范围，例如3-12元环包括3-6元环、3-9元环、5-6元环、5-7元环、6-7元环、6-8元环、和6-10元环等。 Unless otherwise specified, C n-n+m or C n -C n+m includes any specific instance of n to n+m carbons, for example C 1-12 includes C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 , and C 12 , also including any range from n to n+m, for example, C 1-12 includes C 1- 3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12 etc.; similarly, n to n +m means that the number of atoms on the ring is n to n+m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, and a 9-membered ring , 10-membered rings, 11-membered rings, and 12-membered rings, also including any range from n to n+m, for example, 3-12-membered rings include 3-6-membered rings, 3-9-membered rings, 5-6-membered rings ring, 5-7-membered ring, 6-7-membered ring, 6-8-membered ring, and 6-10-membered ring, etc.
术语“离去基团”是指可以被另一种官能团或原子通过取代反应(例如亲核取代反应)所取代的官能团或原子。例如，代表性的离去基团包括三氟甲磺酸酯；氯、溴、碘；磺酸酯基，如甲磺酸酯、甲苯磺酸酯、对溴苯磺酸酯、对甲苯磺酸酯等；酰氧基，如乙酰氧基、三氟乙酰氧基等等。The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (eg, a nucleophilic substitution reaction). For example, representative leaving groups include triflate; chlorine, bromine, iodine; sulfonate groups such as mesylate, tosylate, brosylate, tosylate esters, etc.; acyloxy groups such as acetoxy, trifluoroacetoxy, and the like.
术语“保护基”包括但不限于“氨基保护基”、“羟基保护基”或“巯基保护基”。术语“氨基保护基”是指适合用于阻止氨基氮位上副反应的保护基团。代表性的氨基保护基包括但不限于：甲酰基；酰基，例如链烷酰基(如乙酰基、三氯乙酰基或三氟乙酰基)；烷氧基羰基，如叔丁氧基羰基(Boc)；芳基甲氧羰基，如苄氧羰基(Cbz)和9-芴甲氧羰基(Fmoc)；芳基甲基，如苄基(Bn)、三苯甲基(Tr)、1,1-二-(4'-甲氧基苯基)甲基；甲硅烷基，如三甲基甲硅烷基(TMS)和叔丁基二甲基甲硅烷基(TBS)等等。术语“羟基保护基”是指适合用于阻止羟基副反应的保护基。代表性羟基保护基包括但不限于：烷基，如甲基、乙基和叔丁基；酰基，例如链烷酰基(如乙酰基)；芳基甲基，如苄基(Bn)，对甲氧基苄基(PMB)、9-芴基甲基(Fm)和二苯基甲基(二苯甲基，DPM)；甲硅烷基，如三甲基甲硅烷基(TMS)和叔丁基二甲基甲硅烷基(TBS)等等。The term "protecting group" includes, but is not limited to, "amino protecting group", "hydroxyl protecting group" or "mercapto protecting group". The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (such as acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, such as tert-butoxycarbonyl (Boc) ; arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethyloxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-di -(4'-methoxyphenyl)methyl; silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS) and the like. The term "hydroxyl protecting group" refers to a protecting group suitable for preventing side reactions of the hydroxy group. Representative hydroxy protecting groups include, but are not limited to: alkyl groups such as methyl, ethyl, and tert-butyl; acyl groups such as alkanoyl (such as acetyl); arylmethyl groups such as benzyl (Bn), p-formyl Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyl Dimethylsilyl (TBS) and the like.
本发明的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选 的实施方式包括但不限于本发明的实施例。The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and the methods well known to those skilled in the art Equivalent alternatives, preferred embodiments include but are not limited to the examples of the present invention.
本发明的化合物可以通过本领域技术人员所熟知的常规方法来确认结构，如果本发明涉及化合物的绝对构型，则该绝对构型可以通过本领域常规技术手段予以确证。例如单晶X射线衍射法(SXRD)，把培养出的单晶用Bruker D8 venture衍射仪收集衍射强度数据，光源为CuKα辐射，扫描方式：
Figure PCTCN2022111883-appb-000037
扫描，收集相关数据后，进一步采用直接法(Shelxs97)解析晶体结构，便可以确证绝对构型。 The structure of the compounds of the present invention can be confirmed by conventional methods known to those skilled in the art. If the present invention involves the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, in single crystal X-ray diffraction (SXRD), the cultured single crystal is collected with a Bruker D8 venture diffractometer to collect diffraction intensity data, the light source is CuKα radiation, and the scanning method is:
Figure PCTCN2022111883-appb-000037
After scanning and collecting relevant data, the absolute configuration can be confirmed by further analyzing the crystal structure by direct method (Shelxs97).
本发明所使用的溶剂可经市售获得。本发明采用下述缩略词：aq代表水；HATU代表O-(7-氮杂苯并三唑-1-基)-N,N,N',N'-四甲基脲六氟膦酸盐；EDC代表N-(3-二甲基氨基丙基)-N'-乙基碳二亚胺盐酸盐；m-CPBA代表3-氯过氧苯甲酸；eq代表当量、等量；CDI代表羰基二咪唑；DCM代表二氯甲烷；PE代表石油醚；DIAD代表偶氮二羧酸二异丙酯；DMF代表N,N-二甲基甲酰胺；DMSO代表二甲亚砜；EtOAc代表乙酸乙酯；EtOH代表乙醇；MeOH代表甲醇；CBz代表苄氧羰基，是一种胺保护基团；BOC代表叔丁氧羰基是一种胺保护基团；HOAc代表乙酸；NaCNBH 3代表氰基硼氢化钠；r.t.代表室温；O/N代表过夜；THF代表四氢呋喃；Boc 2O代表二-叔丁基二碳酸酯；TFA代表三氟乙酸；DIPEA代表二异丙基乙基胺；SOCl 2代表氯化亚砜；CS 2代表二硫化碳；TsOH代表对甲苯磺酸；NFSI代表N-氟-N-(苯磺酰基)苯磺酰胺；NCS代表1-氯吡咯烷-2,5-二酮；n-Bu 4NF代表氟化四丁基铵；iPrOH代表2-丙醇；mp代表熔点；LDA代表二异丙基胺基锂。 The solvent used in the present invention is commercially available. The following abbreviations are used in the present invention: aq stands for water; HATU stands for O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphonic acid Salt; EDC stands for N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride; m-CPBA stands for 3-chloroperoxybenzoic acid; eq stands for equivalent, equivalent; CDI DCM stands for dichloromethane; PE stands for petroleum ether; DIAD stands for diisopropyl azodicarboxylate; DMF stands for N,N-dimethylformamide; DMSO stands for dimethylsulfoxide; EtOAc stands for acetic acid EtOH stands for ethanol; MeOH stands for methanol; CBz stands for benzyloxycarbonyl, an amine protecting group; BOC stands for tert-butoxycarbonyl, an amine protecting group; HOAc stands for acetic acid; NaCNBH3 stands for cyanoborohydrogenation Sodium; rt stands for room temperature; O/N stands for overnight; THF stands for tetrahydrofuran; Boc2O stands for di-t-butyldicarbonate; TFA stands for trifluoroacetic acid; DIPEA stands for diisopropylethylamine; SOCl2 stands for chloride sulfoxide; CS 2 represents carbon disulfide; TsOH represents p-toluenesulfonic acid; NFSI represents N-fluoro-N-(benzenesulfonyl)benzenesulfonamide; NCS represents 1-chloropyrrolidine-2,5-dione; n-Bu 4 NF stands for tetrabutylammonium fluoride; iPrOH stands for 2-propanol; mp stands for melting point; LDA stands for lithium diisopropylamide.
化合物依据本领域常规命名原则或者使用
Figure PCTCN2022111883-appb-000038
软件命名，市售化合物采用供应商目录名称。 Compounds are named according to the conventional naming principles in this field or using
Figure PCTCN2022111883-appb-000038
The software is named, and the commercially available compounds adopt the supplier catalog name.
具体实施方式Detailed ways
下面通过实施例对本发明进行详细描述，但并不意味着对本发明任何不利限制。本文已经详细地描述了本发明，其中也公开了其具体实施例方式，对本领域的技术人员而言，在不脱离本发明精神和范围的情况下针对本发明具体实施方式进行各种变化和改进将是显而易见的。The present invention will be described in detail through examples below, but it does not imply any unfavorable limitation to the present invention. The present invention has been described in detail herein, and its specific embodiments are also disclosed. For those skilled in the art, various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention. will be obvious.
实施例1Example 1
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000039
Figure PCTCN2022111883-appb-000039
第一步first step
氮气保护下,将化合物1-1(2.20g,10.6mmol)溶于无水甲醇(20mL)中,加入碳酸钾(2.92g,21.1mmol)和1-2(2.43g,12.7mmol),反应液在25℃下搅拌12小时,向反应液中加入水(40mL),用乙酸乙酯(50mL x 1)萃取。有机相用饱和食盐水洗涤(50mLx 1),无水硫酸钠干燥,过滤,减压浓缩,剩余物用硅胶柱色谱法分离纯化(50：1石油醚/乙酸乙酯),得到化合物1-3。 1H NMR(400MHz,CDCl 3)δ7.56-7.51(m,2H),7.33- 7.29(m,1H),3.64(s,1H)。 Under nitrogen protection, compound 1-1 (2.20g, 10.6mmol) was dissolved in anhydrous methanol (20mL), potassium carbonate (2.92g, 21.1mmol) and 1-2 (2.43g, 12.7mmol) were added, and the reaction solution After stirring at 25°C for 12 hours, water (40 mL) was added to the reaction solution, and extracted with ethyl acetate (50 mL x 1). The organic phase was washed with saturated brine (50mL x 1), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (50:1 petroleum ether/ethyl acetate) to obtain compound 1-3 . 1 H NMR (400 MHz, CDCl 3 ) δ 7.56-7.51 (m, 2H), 7.33-7.29 (m, 1H), 3.64 (s, 1H).
第二步second step
氮气保护下将化合物1-3(1.38g,6.75mmol)溶于无水甲醇(15mL)中,再加入1-4(2.57g,10.1mmol),铜粉(42.9mg,0.675mmol)和碳酸钾(1.86g,13.5mmol),反应液在25℃下搅拌12小时,将反应液旋干,加入乙酸乙酯(50mL),过滤,滤饼用乙酸乙酯(10mL)洗涤,将滤液减压浓缩,剩余物用硅胶柱色谱法分离纯化(20：1石油醚/乙酸乙酯)得到化合物1-5。 1H NMR(400MHz,CDCl 3)δ7.52-7.50(m,2H),7.36-7.31(m,1H),7.25-7.21(m,1H),5.87(d,J＝18.4Hz,1H),1.25(s,12H)。 Under nitrogen protection, compound 1-3 (1.38g, 6.75mmol) was dissolved in anhydrous methanol (15mL), then 1-4 (2.57g, 10.1mmol), copper powder (42.9mg, 0.675mmol) and potassium carbonate were added (1.86g, 13.5mmol), the reaction solution was stirred at 25°C for 12 hours, the reaction solution was spin-dried, added ethyl acetate (50mL), filtered, the filter cake was washed with ethyl acetate (10mL), and the filtrate was concentrated under reduced pressure , the residue was separated and purified by silica gel column chromatography (20:1 petroleum ether/ethyl acetate) to obtain compound 1-5. 1 H NMR (400MHz, CDCl 3 ) δ7.52-7.50(m, 2H), 7.36-7.31(m, 1H), 7.25-7.21(m, 1H), 5.87(d, J=18.4Hz, 1H), 1.25(s,12H).
第三步third step
将化合物1-6(20.0g,117mmol)溶于无水二氯甲烷(100mL)中,加入三甲基氰硅烷(34.8g,350mmol)和三乙胺(17.7g,175mmol)。反应液在20℃下搅拌反应12小时。向反应液加入水(100mL),用二氯甲烷(100mL x 1)萃取。合并有机相,用饱和食盐水洗涤(100mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(3:1石油醚/乙酸乙酯)得到化合物1-7。 1H NMR(400MHz,CDCl 3)δ5.37(s,1H),4.36(d,J＝6.8Hz,2H),4.07(d,J＝6.8Hz,2H),1.45(s,9H)。 Compound 1-6 (20.0 g, 117 mmol) was dissolved in anhydrous dichloromethane (100 mL), and trimethylsilyl cyanide (34.8 g, 350 mmol) and triethylamine (17.7 g, 175 mmol) were added. The reaction solution was stirred and reacted at 20° C. for 12 hours. Water (100 mL) was added to the reaction solution, and extracted with dichloromethane (100 mL x 1). The organic phases were combined, washed with saturated brine (100 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (3:1 petroleum ether/ethyl acetate) to obtain compound 1-7. 1 H NMR (400MHz, CDCl 3 ) δ 5.37 (s, 1H), 4.36 (d, J=6.8Hz, 2H), 4.07 (d, J=6.8Hz, 2H), 1.45 (s, 9H).
第四步the fourth step
将化合物1-7(10.0g,50.5mmol)溶于无水甲醇(100mL)中,加入氯化亚砜(18.0g,151mmol)。反应液在30℃下搅拌反应12小时。反应液减压浓缩。剩余物为化合物1-8。 1H NMR(400MHz,D 2O)δ4.48(d,J＝12.4Hz,2H),4.14(d,J＝12.4Hz,2H),3.84(s,3H)。 Compound 1-7 (10.0 g, 50.5 mmol) was dissolved in anhydrous methanol (100 mL), and thionyl chloride (18.0 g, 151 mmol) was added. The reaction solution was stirred and reacted at 30° C. for 12 hours. The reaction solution was concentrated under reduced pressure. The remainder was compound 1-8. 1 H NMR (400MHz, D 2 O) δ 4.48 (d, J = 12.4Hz, 2H), 4.14 (d, J = 12.4Hz, 2H), 3.84 (s, 3H).
第五步the fifth step
将化合物1-8(8.00g,47.7mmol)溶于甲醇(80mL)中,加入二叔丁基二碳酸酯(19.3g,191mmol)和三乙胺(11.5g,52.5mmol)。反应液在氮气保护下70℃搅拌反应12小时。向反应液加入水(100mL),用乙酸乙酯(100mL x 3)萃取。合并有机相,用饱和食盐水洗涤(50mL x 2),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(3:1石油醚/乙酸乙酯)得到化合物1-9。 1H NMR(400MHz,CDCl 3)δ4.26(d,J＝10.0Hz,2H),4.01(d,J＝10.0Hz,2H),3.90(s,3H),3.72(s,1H),1.45(s,9H)。 Compound 1-8 (8.00g, 47.7mmol) was dissolved in methanol (80mL), and di-tert-butyldicarbonate (19.3g, 191mmol) and triethylamine (11.5g, 52.5mmol) were added. The reaction solution was stirred and reacted at 70° C. for 12 hours under the protection of nitrogen. Water (100 mL) was added to the reaction solution, and extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (3:1 petroleum ether/ethyl acetate) to obtain compound 1-9. 1 H NMR (400MHz, CDCl 3 ) δ4.26(d, J=10.0Hz, 2H), 4.01(d, J=10.0Hz, 2H), 3.90(s, 3H), 3.72(s, 1H), 1.45 (s,9H).
第六步step six
将化合物1-9(2.20g,9.51mmol)溶于无水四氢呋喃(20mL)中,在0℃下分批加入钠氢(571mg,14.3mmol,60％纯度),在该温度下搅拌反应30分钟。然后向反应液中加入1-10(2.09g,9.51mmol),反应液在20℃下搅拌反应2小时。向反应液加入水(10mL),用乙酸乙酯(10mL x 3)萃取。合并有机相,用饱和食盐水洗涤(10mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(5:1石油醚/乙酸乙酯)得到化合物1-11。 1H NMR(400MHz,CDCl 3)δ8.04(s,1H),7.55(d,J＝8.8Hz,1H),6.30(d,J＝7.6Hz,1H),4.48(d,J＝9.6Hz,2H),4.22(d,J＝9.6Hz,2H),3.82(s,3H),1.45(s,9H)。MS-ESI计算值[M-Boc+H] +331和333,实测值331和333。 Compound 1-9 (2.20g, 9.51mmol) was dissolved in anhydrous tetrahydrofuran (20mL), sodium hydrogen (571mg, 14.3mmol, 60% purity) was added in portions at 0°C, and the reaction was stirred at this temperature for 30 minutes . Then 1-10 (2.09g, 9.51mmol) was added to the reaction solution, and the reaction solution was stirred and reacted at 20°C for 2 hours. Water (10 mL) was added to the reaction solution, and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (5:1 petroleum ether/ethyl acetate) to obtain compound 1-11. 1 H NMR (400MHz, CDCl 3 ) δ8.04(s, 1H), 7.55(d, J=8.8Hz, 1H), 6.30(d, J=7.6Hz, 1H), 4.48(d, J=9.6Hz , 2H), 4.22 (d, J=9.6Hz, 2H), 3.82 (s, 3H), 1.45 (s, 9H). MS-ESI calculated [M-Boc+H] + 331 and 333, found 331 and 333.
第七步step seven
将化合物1-11(1.00g,2.32mmol),铁粉(389mg,6.96mmol)和氯化铵(372mg,6.96mmol)加入到乙醇(10mL)和水(2mL)中。反应液在氮气保护下80℃搅拌反应12小时。反应液过滤,向滤液加入水(30mL),用乙酸乙酯(30mL x 2)萃取。合并有机相,用饱和食盐水洗涤(30mL x 2),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(2:1石油醚/乙酸乙酯)得到化合物1-12。 1H NMR(400MHz,CDCl 3)δ8.19(s,1H),7.15(d,J＝8.8Hz,1H),7.00-6.96(m,2H),4.46(d,J＝9.6Hz,2H),4.09(d,J＝9.6Hz,2H),1.48(s,9H)。MS-ESI计算值[M-Boc+H] +269和271,实测值269和271。 Compound 1-11 (1.00 g, 2.32 mmol), iron powder (389 mg, 6.96 mmol) and ammonium chloride (372 mg, 6.96 mmol) were added to ethanol (10 mL) and water (2 mL). The reaction solution was stirred and reacted at 80° C. for 12 hours under the protection of nitrogen. The reaction solution was filtered, water (30 mL) was added to the filtrate, and extracted with ethyl acetate (30 mL x 2). The organic phases were combined, washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (2:1 petroleum ether/ethyl acetate) to obtain compound 1-12. 1 H NMR (400MHz, CDCl 3 ) δ8.19(s, 1H), 7.15(d, J=8.8Hz, 1H), 7.00-6.96(m, 2H), 4.46(d, J=9.6Hz, 2H) , 4.09 (d, J=9.6Hz, 2H), 1.48 (s, 9H). MS-ESI calculated [M-Boc+H] + 269 and 271, found 269 and 271.
第八步eighth step
将化合物1-12(500mg,1.35mmol)溶于无水四氢呋喃(5mL)中,在0℃下滴加硼烷四氢呋喃(1M,5mL,4.06mmol),然后反应液在50℃下搅拌反应1小时。反应液冷却,20℃搅拌下向反应液滴加甲醇(20mL),反应液减压浓缩。剩余物用硅胶柱色谱法分离纯化(2:1石油醚/乙酸乙酯)得到化合物1-13。 1H NMR(400MHz,CDCl 3)δ6.78-6.69(m,3H),4.01(d,J＝9.6Hz,2H),3.90-3.87(m,3H),3.42(s,2H),1.46(s,9H)。MS-ESI计算值[M-56+H] +299和301,实测值299和301。 Compound 1-12 (500mg, 1.35mmol) was dissolved in anhydrous tetrahydrofuran (5mL), borane tetrahydrofuran (1M, 5mL, 4.06mmol) was added dropwise at 0°C, and the reaction solution was stirred at 50°C for 1 hour . The reaction solution was cooled, and methanol (20 mL) was added dropwise to the reaction solution with stirring at 20°C, and the reaction solution was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (2:1 petroleum ether/ethyl acetate) to obtain compound 1-13. 1 H NMR (400MHz, CDCl 3 ) δ6.78-6.69 (m, 3H), 4.01 (d, J=9.6Hz, 2H), 3.90-3.87 (m, 3H), 3.42 (s, 2H), 1.46 ( s, 9H). MS-ESI calculated [M-56+H] + 299 and 301, found 299 and 301.
第九步Ninth step
将化合物1-13(420mg,1.18mmol)溶于二氯甲烷(5mL)中,加入4-二甲基氨基吡啶(28.9mg,0.236mmol),N,N-二异丙基乙胺(306mg,2.36mmol)和1-14(434mg,1.77mmol)。反应液在氮气保护下40℃搅拌反应12小时。向反应液加入水(5mL),用二氯甲烷(5mL x 2)萃取。合并有机相,用饱和食盐水洗涤(10mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(2:1石油醚/乙酸乙酯)得到化合物1-15。 1H NMR(400MHz,CDCl 3)δ8.14(s,1H),8.07(d,J＝8.0Hz,1H),7.92(d,J＝8.0Hz,1H),7.74-7.65(m,2H),7.15-7.12(m,1H),6.82-6.80(m,1H),4.10-3.87(m,6H),1.46(s,9H)。MS-ESI计算值[M-56+H] +507和509,实测值507和509。 Compound 1-13 (420mg, 1.18mmol) was dissolved in dichloromethane (5mL), and 4-dimethylaminopyridine (28.9mg, 0.236mmol), N,N-diisopropylethylamine (306mg, 2.36mmol) and 1-14 (434mg, 1.77mmol). The reaction solution was stirred and reacted at 40° C. for 12 hours under the protection of nitrogen. Water (5 mL) was added to the reaction solution, and extracted with dichloromethane (5 mL x 2). The organic phases were combined, washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (2:1 petroleum ether/ethyl acetate) to obtain compound 1-15. 1 H NMR (400MHz, CDCl 3 ) δ8.14(s, 1H), 8.07(d, J=8.0Hz, 1H), 7.92(d, J=8.0Hz, 1H), 7.74-7.65(m, 2H) ,7.15-7.12(m,1H),6.82-6.80(m,1H),4.10-3.87(m,6H),1.46(s,9H). MS-ESI calculated [M-56+H] + 507 and 509, found 507 and 509.
第十步tenth step
将化合物1-15(200mg,0.355mmol)溶于甲醇(5mL)中,加入盐酸甲醇(4M,5mL,20.0mmol)。反应液在25℃搅拌反应1小时。向反应液加入饱和碳酸氢钠水溶液(10mL),用乙酸乙酯(10mL x 3)萃取。合并有机相,用饱和食盐水洗涤(10mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物为化合物1-16。MS-ESI计算值[M+H] +463和465,实测值463和465。 Compound 1-15 (200 mg, 0.355 mmol) was dissolved in methanol (5 mL), and methanol hydrochloride (4M, 5 mL, 20.0 mmol) was added. The reaction solution was stirred and reacted at 25° C. for 1 hour. Saturated aqueous sodium bicarbonate (10 mL) was added to the reaction solution, and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The remainder was compound 1-16. MS-ESI calculated [M+H] + 463 and 465, found 463 and 465.
第十一步Eleventh step
将化合物1-16(80.0mg,0.135mmol)溶于乙腈(1mL)中,加入碳酸钾(56.0mg,0.405mmol),碘化钾(67.2mg,0.405mmol)和1-17(20.6mg,0.135mmol)。反应液在90℃搅拌反应2小时。向反应液加入水(10mL),用二氯甲烷(10mL x 2)萃取。合并有机相,用饱和食盐水洗涤(10mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用制备TLC板分离纯化(1:1石油醚/乙酸乙酯)得到化合物1-18。MS-ESI计算值 [M+H] +535和537,实测值535和537。 Compound 1-16 (80.0mg, 0.135mmol) was dissolved in acetonitrile (1mL), potassium carbonate (56.0mg, 0.405mmol), potassium iodide (67.2mg, 0.405mmol) and 1-17 (20.6mg, 0.135mmol) were added . The reaction solution was stirred and reacted at 90° C. for 2 hours. Water (10 mL) was added to the reaction solution, and extracted with dichloromethane (10 mL x 2). The organic phases were combined, washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by preparative TLC plate (1:1 petroleum ether/ethyl acetate) to obtain compound 1-18. MS-ESI calculated [M+H] + 535 and 537, found 535 and 537.
第十二步Twelfth step
将化合物1-18(70.0mg,0.130mmol),1-5(51.8mg,0.156mmol),磷酸钾(82.7mg,0.390mmol)和1,1-双(二苯基膦)二茂铁氯化钯(9.51mg,0.0130mmol)加入到二氧六环(2mL)和水(0.4mL)中。反应液在氮气保护下90℃搅拌反应12小时。向反应液中加入水(10mL),用乙酸乙酯(20mL x 2)萃取。合并有机相,用饱和食盐水洗涤(20mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用制备TLC板分离纯化(1:1石油醚/乙酸乙酯)得到化合物1-19。 1H NMR(400MHz,CDCl 3)δ8.18(s,1H),8.13(d,J＝8.0Hz,1H),7.88(d,J＝8.0Hz,1H),7.72-7.68(m,2H),7.64-7.62(m,2H),7.34-7.30(m,1H),7.19-7.16(m,1H),6.94-6.90(m,2H),6.76-6.72(m,1H),4.13(s,2H),3.75(s,3H),3.69(d,J＝8.8Hz,2H),3.42(s,2H),3.33(d,J＝8.8Hz,2H)。MS-ESI计算值[M+H] +661,实测值661。 Compound 1-18 (70.0mg, 0.130mmol), 1-5 (51.8mg, 0.156mmol), potassium phosphate (82.7mg, 0.390mmol) and 1,1-bis(diphenylphosphino)ferrocene were chlorinated Palladium (9.51 mg, 0.0130 mmol) was added to dioxane (2 mL) and water (0.4 mL). The reaction solution was stirred and reacted at 90° C. for 12 hours under the protection of nitrogen. Water (10 mL) was added to the reaction solution, and extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by preparative TLC plate (1:1 petroleum ether/ethyl acetate) to obtain compound 1-19. 1 H NMR (400MHz, CDCl 3 ) δ8.18(s, 1H), 8.13(d, J=8.0Hz, 1H), 7.88(d, J=8.0Hz, 1H), 7.72-7.68(m, 2H) ,7.64-7.62(m,2H),7.34-7.30(m,1H),7.19-7.16(m,1H),6.94-6.90(m,2H),6.76-6.72(m,1H),4.13(s, 2H), 3.75(s, 3H), 3.69(d, J=8.8Hz, 2H), 3.42(s, 2H), 3.33(d, J=8.8Hz, 2H). MS-ESI calculated [M+H] + 661, found 661.
第十三步Step Thirteen
将化合物1-19(60.0mg,0.0908mmol)和一水合氢氧化锂(7.6mg,0.182mmol)加入到四氢呋喃(2mL)和水(0.5mL)中。反应液在氮气保护下20℃搅拌反应12小时。反应液减压浓缩。剩余物经高效液相色谱法(中性,碳酸氢铵体系)分离纯化得到化合物1。 1H NMR(400MHz,CD 3OD)δ8.28-8.24(m,2H),8.06(d,J＝8.0Hz,1H),7.90-7.86(m,1H),7.75-7.70(m,2H),7.62-7.61(m,1H),7.47-7.43(m,1H),7.28-7.26(m,1H),7.07-7.05(m,1H),6.93-6.89(m,1H),6.73-6.69(m,1H),4.36(d,J＝12.0Hz,2H),4.27(s,2H),4.13(d,J＝12.0Hz,2H),3.86(s,2H)。MS-ESI计算值[M+H] +647,实测值647。 Compound 1-19 (60.0 mg, 0.0908 mmol) and lithium hydroxide monohydrate (7.6 mg, 0.182 mmol) were added to tetrahydrofuran (2 mL) and water (0.5 mL). The reaction solution was stirred and reacted at 20° C. for 12 hours under the protection of nitrogen. The reaction solution was concentrated under reduced pressure. The residue was separated and purified by high performance liquid chromatography (neutral, ammonium bicarbonate system) to obtain compound 1. 1 H NMR (400MHz, CD 3 OD) δ8.28-8.24 (m, 2H), 8.06 (d, J = 8.0Hz, 1H), 7.90-7.86 (m, 1H), 7.75-7.70 (m, 2H) ,7.62-7.61(m,1H),7.47-7.43(m,1H),7.28-7.26(m,1H),7.07-7.05(m,1H),6.93-6.89(m,1H),6.73-6.69( m, 1H), 4.36(d, J=12.0Hz, 2H), 4.27(s, 2H), 4.13(d, J=12.0Hz, 2H), 3.86(s, 2H). MS-ESI calculated [M+H] + 647, found 647.
实施例2Example 2
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000040
Figure PCTCN2022111883-appb-000040
第一步first step
参照实施例1第十一步得到化合物2-2。MS-ESI计算值[M+H] +563和565,实测值563和565。 Referring to the eleventh step of Example 1, compound 2-2 was obtained. MS-ESI calculated [M+H] + 563 and 565, found 563 and 565.
第二步second step
参照实施例1第十二步得到化合物2-3。 1H NMR(400MHz,CDCl 3)δ8.17-8.12(m,2H),7.88(d,J＝8.0Hz,1H),7.72-7.62(m,4H),7.34-7.30(m,1H),7.18-7.15(m,1H),6.94-6.90(m,2H),6.75-6.71(m,1H),4.20-4.14(m,2H),4.08(s,2H),3.52(d,J＝8.8Hz,2H),3.15(d,J＝8.8Hz,2H),2.89-2.85(m,2H),2.42-2.38(m,2H),1.31-1.27(m,3H)。MS-ESI计算值[M+H] +689,实测值689。 Referring to the twelfth step of Example 1, compound 2-3 was obtained. 1 H NMR (400MHz, CDCl 3 ) δ8.17-8.12 (m, 2H), 7.88 (d, J=8.0Hz, 1H), 7.72-7.62 (m, 4H), 7.34-7.30 (m, 1H), 7.18-7.15(m,1H),6.94-6.90(m,2H),6.75-6.71(m,1H),4.20-4.14(m,2H),4.08(s,2H),3.52(d,J＝8.8 Hz, 2H), 3.15(d, J=8.8Hz, 2H), 2.89-2.85(m, 2H), 2.42-2.38(m, 2H), 1.31-1.27(m, 3H). MS-ESI calculated [M+H] + 689, found 689.
第三步third step
参照实施例1第十三步得到化合物2。 1H NMR(400MHz,CD 3OD)δ8.26-8.24(m,2H),8.05(d,J＝8.0Hz,1H),7.89-7.85(m,1H),7.76-7.70(m,2H),7.65-7.64(m,1H),7.47-7.43(m,1H),7.25-7.22(m,1H),7.02-7.00(m,1H),6.93-6.89(m,1H),6.73-6.69(m,1H),4.22(s,2H),3.95(d,J＝8.4Hz,2H),3.65(d,J＝8.4Hz,2H),3.17-3.13(m,2H),2.47-2.43(m,2H)。MS-ESI计算值[M+H] +661,实测值661。 Referring to the thirteenth step of Example 1, Compound 2 was obtained. 1 H NMR (400MHz, CD 3 OD) δ8.26-8.24 (m, 2H), 8.05 (d, J = 8.0Hz, 1H), 7.89-7.85 (m, 1H), 7.76-7.70 (m, 2H) ,7.65-7.64(m,1H),7.47-7.43(m,1H),7.25-7.22(m,1H),7.02-7.00(m,1H),6.93-6.89(m,1H),6.73-6.69( m,1H),4.22(s,2H),3.95(d,J=8.4Hz,2H),3.65(d,J=8.4Hz,2H),3.17-3.13(m,2H),2.47-2.43(m ,2H). MS-ESI calculated [M+H] + 661, found 661.
实施例3Example 3
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000041
Figure PCTCN2022111883-appb-000041
第一步first step
将化合物1-16(200mg,0.135mmol)溶于乙腈(3mL)中,加入碳酸钾(121mg,0.875mmol),碘化钾(145mg,0.875mmol)和3-1(52.8mg,0.292mmol)。反应液在90℃搅拌反应12小时。向反应液加入水(10mL),用二氯甲烷(10mL x 2)萃取。合并有机相,用饱和食盐水洗涤(10mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用制备TLC板分离纯化(1:1石油醚/乙酸乙酯)得到化合物3-2。MS-ESI计算值[M+H] +563和565,实测值563和565。 Compound 1-16 (200 mg, 0.135 mmol) was dissolved in acetonitrile (3 mL), potassium carbonate (121 mg, 0.875 mmol), potassium iodide (145 mg, 0.875 mmol) and 3-1 (52.8 mg, 0.292 mmol) were added. The reaction solution was stirred and reacted at 90° C. for 12 hours. Water (10 mL) was added to the reaction solution, and extracted with dichloromethane (10 mL x 2). The organic phases were combined, washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by preparative TLC plate (1:1 petroleum ether/ethyl acetate) to obtain compound 3-2. MS-ESI calculated [M+H] + 563 and 565, found 563 and 565.
第二步second step
将化合物3-2(90.0mg,0.127mmol),1-5(50.6mg,0.152mmol),磷酸钾(80.7mg,0.380mmol)和1,1-双(二苯基膦)二茂铁氯化钯(9.3mg,0.0127mmol)加入到二氧六环(2mL)和水(0.4mL)中。反应液在氮气保护下80℃搅拌反应12小时。反应液减压浓缩。剩余物经高效液相色谱法(中性,碳酸氢铵体系)分离纯化得到化合物3。 1H NMR(400MHz,CD 3OD)δ8.25-8.23(m,2H),8.02(d,J＝8.0Hz,1H),7.87-7.83(m,1H),7.74-7.69(m,2H),7.65-7.64(m,1H),7.46-7.42(m,1H),7.22-7.19(m,1H),6.97(d,J＝8.0Hz,1H),6.92-6.87(m,1H),6.72-6.68(m,1H),4.18(s,2H),3.75(d,J＝8.4Hz,2H),3.43(d,J＝8.4Hz,2H),3.83-3.80(m,2H),2.35-2.32(m,2H),1.76-1.69(m,2H)。MS-ESI计算值[M+H] +675,实测值675。 Compound 3-2 (90.0mg, 0.127mmol), 1-5 (50.6mg, 0.152mmol), potassium phosphate (80.7mg, 0.380mmol) and 1,1-bis(diphenylphosphino)ferrocene were chlorinated Palladium (9.3 mg, 0.0127 mmol) was added to dioxane (2 mL) and water (0.4 mL). The reaction solution was stirred and reacted at 80° C. for 12 hours under the protection of nitrogen. The reaction solution was concentrated under reduced pressure. The residue was separated and purified by high performance liquid chromatography (neutral, ammonium bicarbonate system) to obtain compound 3. 1 H NMR (400MHz, CD 3 OD) δ8.25-8.23 (m, 2H), 8.02 (d, J = 8.0Hz, 1H), 7.87-7.83 (m, 1H), 7.74-7.69 (m, 2H) ,7.65-7.64(m,1H),7.46-7.42(m,1H),7.22-7.19(m,1H),6.97(d,J=8.0Hz,1H),6.92-6.87(m,1H),6.72 -6.68(m,1H),4.18(s,2H),3.75(d,J=8.4Hz,2H),3.43(d,J=8.4Hz,2H),3.83-3.80(m,2H),2.35- 2.32(m,2H),1.76-1.69(m,2H). MS-ESI calculated [M+H] +675 , found 675.
实施例4Example 4
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000042
Figure PCTCN2022111883-appb-000042
第一步first step
将化合物4-1(10.0g,69.4mmol)溶于无水二氯甲烷(100mL)中,0℃下在氮气保护下向反应液中加入三乙胺(20.1g,0.208mol)和甲烷磺酰氯(15.9g,0.139mol)。反应液在25℃下搅拌2小时。向反应液中加入饱和碳酸氢钠(200mL),用乙酸乙酯萃取(100mL x 3),合并有机相,无水硫酸钠干燥,过滤,母液浓缩,得到化合物4-2。 1H NMR(400MHz,CD 3OD)δ4.27(m,2H),4.12-4.09(m,2H),3.02-3.01(m,3H),1.38-1.36(m,2H),1.21-1.18(m,3H),0.99-0.96(m,2H)。 Compound 4-1 (10.0g, 69.4mmol) was dissolved in anhydrous dichloromethane (100mL), and triethylamine (20.1g, 0.208mol) and methanesulfonyl chloride were added to the reaction solution at 0°C under nitrogen protection (15.9 g, 0.139 mol). The reaction solution was stirred at 25°C for 2 hours. Saturated sodium bicarbonate (200 mL) was added to the reaction liquid, extracted with ethyl acetate (100 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated to obtain compound 4-2. 1 H NMR (400MHz, CD 3 OD) δ4.27 (m, 2H), 4.12-4.09 (m, 2H), 3.02-3.01 (m, 3H), 1.38-1.36 (m, 2H), 1.21-1.18 ( m,3H), 0.99-0.96(m,2H).
第二步second step
参照实施例1第十一步得到化合物4-3。MS-ESI计算值[M+H] +589和591,实测值589和591。 Referring to the eleventh step of Example 1, compound 4-3 was obtained. MS-ESI calculated [M+H] + 589 and 591, found 589 and 591.
第三步third step
参照实施例1第十二步得到化合物4-4。 1H NMR(400MHz,CDCl 3)δ8.16-8.11(m,2H),7.85(d,J＝8.0Hz,1H),7.71-7.61(m,4H),7.34-7.30(m,1H),7.16-7.14(m,1H),6.90-6.88(m,2H),6.74-6.70(m,1H),4.16-4.09(m,4H),3.79(s,2H),3.54(d,J＝8.8Hz,2H),3.21(d,J＝8.8Hz,2H),3.09-3.05(m,2H),2.80(s,2H),1.21-1.19(m,3H)。MS-ESI计算值[M+H] +715,实测值715。 Referring to the twelfth step of Example 1, compound 4-4 was obtained. 1 H NMR (400MHz, CDCl 3 ) δ8.16-8.11 (m, 2H), 7.85 (d, J=8.0Hz, 1H), 7.71-7.61 (m, 4H), 7.34-7.30 (m, 1H), 7.16-7.14(m,1H),6.90-6.88(m,2H),6.74-6.70(m,1H),4.16-4.09(m,4H),3.79(s,2H),3.54(d,J＝8.8 Hz, 2H), 3.21 (d, J=8.8Hz, 2H), 3.09-3.05 (m, 2H), 2.80 (s, 2H), 1.21-1.19 (m, 3H). MS-ESI calculated [M+H] + 715, found 715.
第四步the fourth step
参照实施例1第十三步得到化合物4。 1H NMR(400MHz,CD 3OD)δ8.26-8.22(m,2H),8.02(d,J＝8.0Hz,1H),7.87-7.85(m,1H),7.74-7.68(m,2H),7.63-7.62(m,1H),7.44-7.42(m,1H),7.22-7.21(m,1H),7.02-7.00(m,1H),6.82-6.80(m,1H),6.72-6.67(m,1H),4.25(s,2H),4.19(d,J＝10.8Hz,2H),3.93(d,J＝8.8Hz,2H),3.20(s,2H),1.20-1.17(m,2H),0.82-0.79(m,2H)。MS-ESI计算值[M+H] +687,实测值687。 Referring to the thirteenth step of Example 1, compound 4 was obtained. 1 H NMR (400MHz, CD 3 OD) δ8.26-8.22 (m, 2H), 8.02 (d, J = 8.0Hz, 1H), 7.87-7.85 (m, 1H), 7.74-7.68 (m, 2H) ,7.63-7.62(m,1H),7.44-7.42(m,1H),7.22-7.21(m,1H),7.02-7.00(m,1H),6.82-6.80(m,1H),6.72-6.67( m,1H),4.25(s,2H),4.19(d,J=10.8Hz,2H),3.93(d,J=8.8Hz,2H),3.20(s,2H),1.20-1.17(m,2H ),0.82-0.79(m,2H). MS-ESI calculated [M+H] + 687, found 687.
实施例5Example 5
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000043
Figure PCTCN2022111883-appb-000043
第一步first step
将化合物5-1(1.00g,7.57mmol)溶于二氯乙烷(10mL)中,在0℃下加入戴斯-马丁过碘烷(4.81g,11.4mmol)。反应液在25℃搅拌反应12小时。向反应液加入饱和碳酸氢钠水溶液(50mL),用二氯甲烷(30mL x 1)萃取。合并有机相,用饱和食盐水洗涤(30mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(5:1石油醚/乙酸乙酯)得到化合物5-2。 1H NMR(400MHz,CDCl 3)δ9.65(s,1H), 3.73(s,3H),1.34(s,6H)。 Compound 5-1 (1.00 g, 7.57 mmol) was dissolved in dichloroethane (10 mL), and Dess-Martin periodinane (4.81 g, 11.4 mmol) was added at 0°C. The reaction solution was stirred and reacted at 25° C. for 12 hours. Saturated aqueous sodium bicarbonate (50 mL) was added to the reaction solution, and extracted with dichloromethane (30 mL x 1). The organic phases were combined, washed with saturated brine (30 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (5:1 petroleum ether/ethyl acetate) to obtain compound 5-2. 1 H NMR (400MHz, CDCl 3 ) δ9.65(s, 1H), 3.73(s, 3H), 1.34(s, 6H).
第二步second step
将化合物1-16(100mg,0.216mmol)溶于1,2-二氯乙烷(5mL)中,在25℃下加入四乙氧基钛(98.5mg,0.432mmol)和5-2(56.2mg,0.432mmol)。反应液在25℃搅拌反应0.5小时,然后加入三乙酰氧基硼氢化钠(137mg,0.648mmol),反应液在50℃搅拌反应2小时。向反应液加入水(20mL),用二氯甲烷(20mL x 2)萃取。合并有机相,用饱和食盐水洗涤(20mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用制备TLC板分离纯化(3:1石油醚/乙酸乙酯)得到化合物5-3。MS-ESI计算值[M+H] +577和579,实测值577和579。 Compound 1-16 (100 mg, 0.216 mmol) was dissolved in 1,2-dichloroethane (5 mL), and tetraethoxytitanium (98.5 mg, 0.432 mmol) and 5-2 (56.2 mg ,0.432mmol). The reaction solution was stirred and reacted at 25°C for 0.5 hours, then sodium triacetoxyborohydride (137mg, 0.648mmol) was added, and the reaction solution was stirred and reacted at 50°C for 2 hours. Water (20 mL) was added to the reaction solution, and extracted with dichloromethane (20 mL x 2). The organic phases were combined, washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by preparative TLC plate (3:1 petroleum ether/ethyl acetate) to obtain compound 5-3. MS-ESI calculated [M+H] + 577 and 579, found 577 and 579.
第三步third step
参照实施例1第十二步得到化合物5-4。MS-ESI计算值[M+H] +703,实测值703。 Referring to the twelfth step of Example 1, compound 5-4 was obtained. MS-ESI calculated value [M+H] + 703, found value 703.
第四步the fourth step
参照实施例1第十三步得到化合物5。 1H NMR(400MHz,CD 3OD)δ8.24-8.22(m,2H),8.01(d,J＝8.0Hz,1H),7.87-7.84(m,1H),7.74-7.67(m,3H),7.45-7.43(m,1H),7.20-7.19(m,1H),6.99-6.97(m,1H),6.90-6.87(m,1H),6.72-6.68(m,1H),4.20(s,2H),3.85(d,J＝9.6Hz,2H),3.56(d,J＝9.6Hz,2H),2.92(s,2H),1.17(s,6H)。MS-ESI计算值[M+H] +689,实测值689。 Compound 5 was obtained by referring to the thirteenth step of Example 1. 1 H NMR (400MHz, CD 3 OD) δ8.24-8.22 (m, 2H), 8.01 (d, J = 8.0Hz, 1H), 7.87-7.84 (m, 1H), 7.74-7.67 (m, 3H) ,7.45-7.43(m,1H),7.20-7.19(m,1H),6.99-6.97(m,1H),6.90-6.87(m,1H),6.72-6.68(m,1H),4.20(s, 2H), 3.85(d, J=9.6Hz, 2H), 3.56(d, J=9.6Hz, 2H), 2.92(s, 2H), 1.17(s, 6H). MS-ESI calculated [M+H] + 689, found 689.
实施例6Example 6
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000044
Figure PCTCN2022111883-appb-000044
第一步first step
氮气保护下将联硼酸频那醇酯(819mg,3.23mmol)加入无水四氢呋喃(10mL)中,再加入氯化亚铜(29.0mg,0.293mmol)和4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(170mg,0.293mmol),反应液在25℃下搅拌5分钟,再加入叔丁醇钠(310mg,3.23mmol),反应液在25℃下搅拌5分钟,再加入化合物1-3(600mg, 2.93mmol)和碘甲烷(1.67g,11.7mmol),反应液在25℃下搅拌710分钟,向反应液中加入水(20mL),用乙酸乙酯(20mL x 1)萃取,有机相用饱和食盐水洗涤(20mLx 1),无水硫酸钠干燥,过滤,减压浓缩,剩余物用硅胶柱色谱法分离纯化(10:1石油醚/乙酸乙酯)得到化合物6-1。 1H NMR(400MHz,CDCl 3)δ7.58-7.55(m,2H),7.32-7.27(m,1H),5.26(s,1H),2.28(s,3H),1.32(s,12H)。 Under nitrogen protection, diboronic acid pinacol ester (819mg, 3.23mmol) was added into anhydrous tetrahydrofuran (10mL), then cuprous chloride (29.0mg, 0.293mmol) and 4,5-bis(diphenylphosphine) were added -9,9-Dimethylxanthene (170mg, 0.293mmol), the reaction solution was stirred at 25°C for 5 minutes, then sodium tert-butoxide (310mg, 3.23mmol) was added, and the reaction solution was stirred at 25°C for 5 minutes , then added compound 1-3 (600mg, 2.93mmol) and iodomethane (1.67g, 11.7mmol), the reaction solution was stirred at 25°C for 710 minutes, water (20mL) was added to the reaction solution, and ethyl acetate (20mL x 1) extraction, the organic phase was washed with saturated brine (20mLx 1), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (10:1 petroleum ether/ethyl acetate) to obtain Compound 6-1. 1 H NMR (400 MHz, CDCl 3 ) δ 7.58-7.55 (m, 2H), 7.32-7.27 (m, 1H), 5.26 (s, 1H), 2.28 (s, 3H), 1.32 (s, 12H).
第二步second step
参照实施例1第十二步得到化合物6-2。MS-ESI计算值[M+H] +675,实测值675。 Referring to the twelfth step of Example 1, compound 6-2 was obtained. MS-ESI calculated [M+H] +675 , found 675.
第三步third step
参照实施例1第十三步得到化合物6。 1H NMR(400MHz,CD 3OD)δ8.26-8.21(m,2H),8.06(d,J＝7.6Hz,1H),7.91-7.87(m,1H),7.71(t,J＝8.2Hz,2H),7.50-7.46(m,1H),7.38(d,J＝1.6Hz,1H),7.08-7.02(m,2H),6.19(s,1H),4.39-4.26(m,4H),4.16-4.13(m,2H),3.86(s,2H),1.98(s,3H)。MS-ESI计算值[M+H] +661,实测值661。 Referring to the thirteenth step of Example 1, compound 6 was obtained. 1 H NMR (400MHz, CD 3 OD) δ8.26-8.21(m, 2H), 8.06(d, J=7.6Hz, 1H), 7.91-7.87(m, 1H), 7.71(t, J=8.2Hz ,2H),7.50-7.46(m,1H),7.38(d,J=1.6Hz,1H),7.08-7.02(m,2H),6.19(s,1H),4.39-4.26(m,4H), 4.16-4.13 (m, 2H), 3.86 (s, 2H), 1.98 (s, 3H). MS-ESI calculated [M+H] + 661, found 661.
实施例7Example 7
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000045
Figure PCTCN2022111883-appb-000045
第一步first step
参照实施例1第十二步得到化合物7-1。MS-ESI计算值[M+H] +717,实测值717。 Referring to the twelfth step of Example 1, compound 7-1 was obtained. MS-ESI calculated [M+H] + 717, found 717.
第二步second step
参照实施例1第十三步得到化合物7。 1H NMR(400MHz,CD 3OD)δ8.23-8.19(m,2H),8.03(d,J＝8.0Hz,1H),7.88-7.84(m,1H),7.73-7.69(m,2H),7.49-7.43(m,2H),6.98-6.97(m,2H),6.17(s,1H),4.24-4.22(m,2H),3.85(d,J＝9.6Hz,2H),3.55(d,J＝9.6Hz,2H),2.92(s,2H),1.98(s,3H),1.17(s,6H)。MS-ESI计算值[M+H] +703,实测值703。 Referring to the thirteenth step of Example 1, Compound 7 was obtained. 1 H NMR (400MHz, CD 3 OD) δ8.23-8.19 (m, 2H), 8.03 (d, J = 8.0Hz, 1H), 7.88-7.84 (m, 1H), 7.73-7.69 (m, 2H) ,7.49-7.43(m,2H),6.98-6.97(m,2H),6.17(s,1H),4.24-4.22(m,2H),3.85(d,J＝9.6Hz,2H),3.55(d , J=9.6Hz, 2H), 2.92(s, 2H), 1.98(s, 3H), 1.17(s, 6H). MS-ESI calculated value [M+H] + 703, found value 703.
实施例8Example 8
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000046
Figure PCTCN2022111883-appb-000046
第一步first step
将锌粉(1.81g,27.7mmol)和1,2-二溴乙烷(307mg,1.63mmol)加入到N,N-二甲基甲酰胺(20mL)中。反应液在70℃搅拌反应10分钟。然后反应液冷却到20℃。三甲基氯硅烷(177mg,1.63mmol)滴加到反应液中,然后反应液20℃搅拌反应50分钟。8-2(5.00g,16.3mmol),三(二亚苄基丙酮)二钯(299mg,0.326mmol),三(2-呋喃基)膦(151mg,0.653mmol)加入到反应液中。反应液在氮气保护下40℃搅拌反应1小时。8-1(6.93g,24.5mmol)溶于N,N-二甲基甲酰胺(40mL)加入到反应液中,反应液在氮气保护下60℃搅拌反应10小时。向反应液中加入水(100mL),用乙酸乙酯(100mL x 2)萃取。合并有机相,用饱和食盐水洗涤(100mL x 3),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(5:1石油醚/乙酸乙酯)得到化合物8-3。 1H NMR(400MHz,CDCl 3)δ7.62-7.58(m,2H),7.33-7.29(m,1H),4.60-4.57(m,2H),4.40-4.34(m,1H),4.24-4.20(m,2H),1.48(s,9H)。MS-ESI计算值[M-56+H] +280,实测值280。 Zinc dust (1.81 g, 27.7 mmol) and 1,2-dibromoethane (307 mg, 1.63 mmol) were added to N,N-dimethylformamide (20 mL). The reaction solution was stirred and reacted at 70° C. for 10 minutes. The reaction solution was then cooled to 20°C. Chlorotrimethylsilane (177mg, 1.63mmol) was added dropwise to the reaction solution, and then the reaction solution was stirred at 20°C for 50 minutes. 8-2 (5.00g, 16.3mmol), tris(dibenzylideneacetone)dipalladium (299mg, 0.326mmol), tris(2-furyl)phosphine (151mg, 0.653mmol) were added to the reaction solution. The reaction solution was stirred and reacted at 40° C. for 1 hour under the protection of nitrogen. 8-1 (6.93g, 24.5mmol) was dissolved in N,N-dimethylformamide (40mL) and added to the reaction solution, and the reaction solution was stirred and reacted at 60°C for 10 hours under the protection of nitrogen. Water (100 mL) was added to the reaction solution, and extracted with ethyl acetate (100 mL x 2). The organic phases were combined, washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (5:1 petroleum ether/ethyl acetate) to obtain compound 8-3. 1 H NMR (400MHz, CDCl 3 )δ7.62-7.58(m,2H),7.33-7.29(m,1H),4.60-4.57(m,2H),4.40-4.34(m,1H),4.24-4.20 (m,2H), 1.48(s,9H). MS-ESI calculated value [M-56+H] + 280, measured value 280.
第二步second step
将化合物8-3(1.10g,3.28mmol)溶于甲醇(10mL)中,加入盐酸甲醇(4M,10mL,40.0mmol)。反应液在20℃搅拌反应1小时。反应液减压浓缩。剩余物为化合物8-4的盐酸盐。MS-ESI计算值[M+H] +236,实测值236。 Compound 8-3 (1.10 g, 3.28 mmol) was dissolved in methanol (10 mL), and methanol hydrochloride (4M, 10 mL, 40.0 mmol) was added. The reaction solution was stirred and reacted at 20° C. for 1 hour. The reaction solution was concentrated under reduced pressure. The residue was the hydrochloride salt of compound 8-4. MS-ESI calculated [M+H] + 236, found 236.
第三步third step
将化合物1-18(250mg,0.467mmol),8-4的盐酸盐(140mg,0.514mmol),碳酸铯(456mg,1.40mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(27.0mg,0.0467mmol)和三(二亚苄基丙酮)二钯(21.4mg,0.0234mmol)加 入到二氧六环(5mL)中。反应液在氮气保护下90℃搅拌反应12小时。向反应液中加入水(10mL),用乙酸乙酯(10mL x 3)萃取。合并有机相,用饱和食盐水洗涤(10mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用制备TLC板分离纯化(1:1石油醚/乙酸乙酯)得到化合物8-5。MS-ESI计算值[M+H] +690,实测值690。 Compound 1-18 (250mg, 0.467mmol), 8-4 hydrochloride (140mg, 0.514mmol), cesium carbonate (456mg, 1.40mmol), 4,5-bisdiphenylphosphine-9,9-di Methylxanthene (27.0 mg, 0.0467 mmol) and tris(dibenzylideneacetone)dipalladium (21.4 mg, 0.0234 mmol) were added to dioxane (5 mL). The reaction solution was stirred and reacted at 90° C. for 12 hours under the protection of nitrogen. Water (10 mL) was added to the reaction solution, and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by preparative TLC plate (1:1 petroleum ether/ethyl acetate) to obtain compound 8-5. MS-ESI calculated value [M+H] + 690, found value 690.
第四步the fourth step
将化合物8-5(80.0mg,0.0905mmol)和一水合氢氧化锂(7.6mg,0.181mmol)加入到四氢呋喃(5mL)和水(1mL)中。反应液在氮气保护下20℃搅拌反应12小时。反应液减压浓缩。剩余物经高效液相色谱法(中性,碳酸氢铵体系)分离纯化得到化合物8。 1H NMR(400MHz,CD 3OD)δ8.22-8.19(m,2H),8.04-8.02(m,1H),7.85-7.83(m,1H),7.71-7.66(m,2H),7.45-7.43(m,1H),6.89(d,J＝8.8Hz,1H),6.53-6.52(m,1H),6.31-6.28(m,1H),4.60-4.58(m,1H),4.34-4.27(m,4H),4.22-4.19(m,4H),4.09-4.06(m,2H),3.85(s,2H)。MS-ESI计算值[M+H] +676,实测值676。 Compound 8-5 (80.0 mg, 0.0905 mmol) and lithium hydroxide monohydrate (7.6 mg, 0.181 mmol) were added to tetrahydrofuran (5 mL) and water (1 mL). The reaction solution was stirred and reacted at 20° C. for 12 hours under the protection of nitrogen. The reaction solution was concentrated under reduced pressure. The residue was separated and purified by high performance liquid chromatography (neutral, ammonium bicarbonate system) to obtain compound 8. 1 H NMR (400MHz, CD 3 OD) δ8.22-8.19(m,2H),8.04-8.02(m,1H),7.85-7.83(m,1H),7.71-7.66(m,2H),7.45- 7.43(m,1H),6.89(d,J＝8.8Hz,1H),6.53-6.52(m,1H),6.31-6.28(m,1H),4.60-4.58(m,1H),4.34-4.27( m, 4H), 4.22-4.19 (m, 4H), 4.09-4.06 (m, 2H), 3.85 (s, 2H). MS-ESI calculated [M+H] + 676, found 676.
实施例9Example 9
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000047
Figure PCTCN2022111883-appb-000047
第一步first step
参照实施例6第一步得到化合物9-2。 1H NMR(400MHz,CDCl 3)δ7.17-7.13(m,2H),7.00-6.95(m,1H),5.35(d,J＝0.8Hz,1H),2.28(d,J＝1.2Hz,3H),1.33(s,12H)。 Referring to the first step of Example 6, compound 9-2 was obtained. 1 H NMR (400MHz, CDCl 3 ) δ7.17-7.13(m, 2H), 7.00-6.95(m, 1H), 5.35(d, J=0.8Hz, 1H), 2.28(d, J=1.2Hz, 3H), 1.33(s, 12H).
第二步second step
参照实施例1第十二步得到化合物9-3。MS-ESI计算值[M+H] +625,实测值625。 Referring to the twelfth step of Example 1, compound 9-3 was obtained. MS-ESI calculated [M+H] + 625, found 625.
第三步third step
参照实施例1第十三步得到化合物9。 1H NMR(400MHz,CD 3OD)δ8.26-8.22(m,2H),8.06(d,J＝8.0Hz,1H), 7.91-7.87(m,1H),7.43-7.42(m,1H),7.31-7.28(m,2H),7.13-7.07(m,3H),6.29-6.28(m,1H),4.38-4.35(m,2H),4.29(s,2H),4.14-4.11(m,2H),3.86(s,2H),2.00(d,J＝1.2Hz,3H)。MS-ESI计算值[M+H] +611,实测值611。 Referring to the thirteenth step of Example 1, compound 9 was obtained. 1 H NMR (400MHz, CD 3 OD) δ8.26-8.22(m, 2H), 8.06(d, J=8.0Hz, 1H), 7.91-7.87(m, 1H), 7.43-7.42(m, 1H) ,7.31-7.28(m,2H),7.13-7.07(m,3H),6.29-6.28(m,1H),4.38-4.35(m,2H),4.29(s,2H),4.14-4.11(m, 2H), 3.86(s, 2H), 2.00(d, J=1.2Hz, 3H). MS-ESI calculated value [M+H] + 611, found value 611.
实施例10Example 10
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000048
Figure PCTCN2022111883-appb-000048
第一步first step
将化合物10-1(500mg,2.07mmol),双联频哪醇硼酸酯(580mg,2.28mmol),乙酸钾(611mg,6.22mmol)和1,1-双(二苯基膦)二茂铁氯化钯(152mg,0.207mmol)加入到二氧六环(5mL)中。反应液在氮气保护下80℃搅拌反应12小时。向反应液中加入水(10mL),用乙酸乙酯(10mL x 2)萃取。合并有机相,用饱和食盐水洗涤(10mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(10/1石油醚/乙酸乙酯)得到化合物10-2。MS-ESI计算值[M+H] +289,实测值289。 1H NMR:(400MHz,CDCl 3)δ7.38-7.30(m,2H),6.97-6.94(m,1H),6.73(s,0.25H),6.54(s,0.5H),6.36(s,0.25H),1.35(s,12H)。 Compound 10-1 (500mg, 2.07mmol), bis-pinacol borate (580mg, 2.28mmol), potassium acetate (611mg, 6.22mmol) and 1,1-bis(diphenylphosphino)ferrocene Palladium chloride (152 mg, 0.207 mmol) was added to dioxane (5 mL). The reaction solution was stirred and reacted at 80° C. for 12 hours under the protection of nitrogen. Water (10 mL) was added to the reaction solution, and extracted with ethyl acetate (10 mL x 2). The organic phases were combined, washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (10/1 petroleum ether/ethyl acetate) to obtain compound 10-2. MS-ESI calculated [M+H] + 289, found 289. 1 H NMR: (400MHz, CDCl 3 )δ7.38-7.30(m,2H),6.97-6.94(m,1H),6.73(s,0.25H),6.54(s,0.5H),6.36(s, 0.25H), 1.35(s, 12H).
第二步second step
将化合物1-18(200mg,0.374mmol),10-2(129mg,0.448mmol),磷酸钾(238mg,1.12mmol)和1,1-双(二苯基膦)二茂铁氯化钯(27.3mg,0.0374mmol)加入到1,4-二氧六环(5mL)和水(1mL)中。反应液在氮气保护下80℃搅拌反应12小时。向反应液中加入水(10mL),用乙酸乙酯(10mL x 2)萃取。合并有机相,用饱和食盐水洗涤(10mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用制备薄层层析法板分离纯化(1/1石油醚/乙酸乙酯,Rf＝0.3)得到化合物10-3。MS-ESI计算值[M+H] +617,实测值617。 Compound 1-18 (200mg, 0.374mmol), 10-2 (129mg, 0.448mmol), potassium phosphate (238mg, 1.12mmol) and 1,1-bis(diphenylphosphine)ferrocenepalladium chloride (27.3 mg, 0.0374 mmol) was added to 1,4-dioxane (5 mL) and water (1 mL). The reaction solution was stirred and reacted at 80° C. for 12 hours under the protection of nitrogen. Water (10 mL) was added to the reaction solution, and extracted with ethyl acetate (10 mL x 2). The organic phases were combined, washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by preparative thin layer chromatography (1/1 petroleum ether/ethyl acetate, Rf=0.3) to obtain compound 10-3. MS-ESI calculated [M+H] + 617, found 617.
第三步third step
将化合物10-3(150mg,0.163mmol)和氢氧化钠(13.0mg,0.326mmol)加入到四氢呋喃(5mL)和水(1mL)中。反应液在氮气保护下60℃搅拌反应1小时。反应液减压浓缩。剩余物经高效液相色谱法(中性, 碳酸氢铵体系)分离纯化得到化合物10。 1H NMR(400MHz,CD 3OD)δ8.27-8.22(m,2H),8.02(d,J＝7.6Hz,1H),7.90-7.82(m,1H),7.70(s,1H),7.33-7.30(m,1H),7.12(s,0.25H),7.09-7.03(m,3H),6.95-6.89(m,1.5H),6.75(s,0.25H),4.22(s,2H),3.76-3.74(m,2H),3.37-3.35(m,2H),3.31-3.29(m,2H)。MS-ESI计算值[M+H] +603,实测值603。 Compound 10-3 (150 mg, 0.163 mmol) and sodium hydroxide (13.0 mg, 0.326 mmol) were added to tetrahydrofuran (5 mL) and water (1 mL). The reaction solution was stirred and reacted at 60° C. for 1 hour under the protection of nitrogen. The reaction solution was concentrated under reduced pressure. The residue was separated and purified by high performance liquid chromatography (neutral, ammonium bicarbonate system) to obtain compound 10. 1 H NMR (400MHz, CD 3 OD) δ8.27-8.22 (m, 2H), 8.02 (d, J = 7.6Hz, 1H), 7.90-7.82 (m, 1H), 7.70 (s, 1H), 7.33 -7.30(m,1H),7.12(s,0.25H),7.09-7.03(m,3H),6.95-6.89(m,1.5H),6.75(s,0.25H),4.22(s,2H), 3.76-3.74 (m, 2H), 3.37-3.35 (m, 2H), 3.31-3.29 (m, 2H). MS-ESI calculated value [M+H] + 603, found value 603.
实施例11Example 11
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000049
Figure PCTCN2022111883-appb-000049
第一步first step
将化合物1-15(3.00g,5.32mmol),10-2(1.84g,6.39mmol),磷酸钾(2.26g,10.1mmol)和1,1-双(二苯基膦)二茂铁氯化钯(390mg,0.533mmol)加入到二氧六环(40mL)和水(4mL)中。反应液在氮气保护下80℃搅拌反应12小时。向反应液中加入水(100mL),用乙酸乙酯(100mL x 2)萃取。合并有机相,用饱和食盐水洗涤(100mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(1:1石油醚/乙酸乙酯)得到化合物11-1。 1H NMR:(400MHz,CDCl 3)δ8.18(s,1H),8.09(d,J＝8.0Hz,1H),7.91(d,J＝8.0Hz,1H),7.77-7.70(m,1H),7.66(d,J＝2.0Hz,1H),7.22(dd,J＝2.0,8.4Hz,1H),7.05-6.95(m,3H),6.84(dd,J＝2.0,8.4Hz,1H),6.74(s,0.25H),6.56(s,0.5H),6.37(s,0.25H),4.14-3.90(m,6H),1.48(s,9H)。MS-ESI计算值[M-Boc+H] +545,实测值545。 Compound 1-15 (3.00g, 5.32mmol), 10-2 (1.84g, 6.39mmol), potassium phosphate (2.26g, 10.1mmol) and 1,1-bis(diphenylphosphino)ferrocene were chlorinated Palladium (390 mg, 0.533 mmol) was added to dioxane (40 mL) and water (4 mL). The reaction solution was stirred and reacted at 80° C. for 12 hours under the protection of nitrogen. Water (100 mL) was added to the reaction solution, and extracted with ethyl acetate (100 mL x 2). The organic phases were combined, washed with saturated brine (100 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (1:1 petroleum ether/ethyl acetate) to obtain compound 11-1. 1 H NMR: (400MHz, CDCl 3 ) δ8.18(s, 1H), 8.09(d, J=8.0Hz, 1H), 7.91(d, J=8.0Hz, 1H), 7.77-7.70(m, 1H ),7.66(d,J=2.0Hz,1H),7.22(dd,J=2.0,8.4Hz,1H),7.05-6.95(m,3H),6.84(dd,J=2.0,8.4Hz,1H) ,6.74(s,0.25H),6.56(s,0.5H),6.37(s,0.25H),4.14-3.90(m,6H),1.48(s,9H). MS-ESI calculated value [M-Boc+H] + 545, found value 545.
第二步second step
将化合物11-1(3.40g,5.27mmol)溶于甲醇(50mL)中,加入氯化氢的甲醇溶液(4M,25mL,100mmol)。 反应液在25℃搅拌反应1小时。向反应液加入饱和碳酸氢钠水溶液(200mL),用乙酸乙酯(100mL x 3)萃取。合并有机相,用饱和食盐水洗涤(100mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩得到化合物11-2。 1H NMR:(400MHz,CDCl 3)δ8.19(s,1H),8.11(d,J＝8.0Hz,1H),7.89(d,J＝8.0Hz,1H),7.77-7.62(m,2H),7.19(dd,J＝2.0,8.4Hz,1H),7.02-6.92(m,3H),6.82(dd,J＝2.0,8.4Hz,1H),6.73(s,0.25H),6.55(s,0.5H),6.37(s,0.25H),4.14(s,2H),3.82(d,J＝9.6Hz,2H),3.64(d,J＝9.6Hz,2H)。MS-ESI计算值[M+H] +545,实测值545。 Compound 11-1 (3.40 g, 5.27 mmol) was dissolved in methanol (50 mL), and a methanolic hydrogen chloride solution (4M, 25 mL, 100 mmol) was added. The reaction solution was stirred and reacted at 25° C. for 1 hour. Saturated aqueous sodium bicarbonate (200 mL) was added to the reaction solution, and extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed with saturated brine (100 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 11-2. 1 H NMR: (400MHz, CDCl 3 ) δ8.19(s, 1H), 8.11(d, J=8.0Hz, 1H), 7.89(d, J=8.0Hz, 1H), 7.77-7.62(m, 2H ),7.19(dd,J=2.0,8.4Hz,1H),7.02-6.92(m,3H),6.82(dd,J=2.0,8.4Hz,1H),6.73(s,0.25H),6.55(s , 0.5H), 6.37(s, 0.25H), 4.14(s, 2H), 3.82(d, J=9.6Hz, 2H), 3.64(d, J=9.6Hz, 2H). MS-ESI calculated [M+H] + 545, found 545.
第三步third step
将化合物11-2(600mg,1.10mmol)溶于乙腈(5mL)中,加入碳酸钾(456mg,3.30mmol),碘化钾(548mg,3.30mmol)和2-1(199mg,1.10mmol)。反应液在90℃搅拌反应12小时。向反应液加入水(20mL),用二氯甲烷(20mL x 2)萃取。合并有机相,用饱和食盐水洗涤(20mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(1:1石油醚/乙酸乙酯)得到化合物11-3。MS-ESI计算值[M+H] +645,实测值645。 Compound 11-2 (600 mg, 1.10 mmol) was dissolved in acetonitrile (5 mL), potassium carbonate (456 mg, 3.30 mmol), potassium iodide (548 mg, 3.30 mmol) and 2-1 (199 mg, 1.10 mmol) were added. The reaction solution was stirred and reacted at 90° C. for 12 hours. Water (20 mL) was added to the reaction solution, and extracted with dichloromethane (20 mL x 2). The organic phases were combined, washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (1:1 petroleum ether/ethyl acetate) to obtain compound 11-3. MS-ESI calculated [M+H] + 645, found 645.
第四步the fourth step
将化合物11-3(360mg,0.554mmol)和氢氧化钠(44.3mg,1.11mmol)加入到四氢呋喃(5mL)和水(1mL)中。反应液在氮气保护下60℃搅拌反应12小时。反应液减压浓缩。剩余物经高效液相色谱法(中性,碳酸氢铵体系)分离纯化得到化合物11。 1H NMR:(400MHz,CD 3OD)δ8.28-8.20(m,2H),8.03(d,J＝8.0Hz,1H),7.90-7.82(m,1H),7.62(d,J＝2.0Hz,1H),7.32(dd,J＝2.0,8.4Hz,1H),7.10(s,0.25H),7.08-6.99(m,3H),6.96-6.84(m,1.5H),6.73(s,0.25H),4.23(s,2H),4.00(d,J＝10.4Hz,2H),3.70(d,J＝10.4Hz,2H),3.19(t,J＝6.8Hz,2H),2.47(t,J＝6.8Hz,2H)。MS-ESI计算值[M+H] +617,实测值617。 Compound 11-3 (360 mg, 0.554 mmol) and sodium hydroxide (44.3 mg, 1.11 mmol) were added to tetrahydrofuran (5 mL) and water (1 mL). The reaction solution was stirred and reacted at 60° C. for 12 hours under the protection of nitrogen. The reaction solution was concentrated under reduced pressure. The residue was separated and purified by high performance liquid chromatography (neutral, ammonium bicarbonate system) to obtain compound 11. 1 H NMR: (400MHz, CD 3 OD) δ8.28-8.20 (m, 2H), 8.03 (d, J = 8.0Hz, 1H), 7.90-7.82 (m, 1H), 7.62 (d, J = 2.0 Hz, 1H), 7.32(dd, J=2.0, 8.4Hz, 1H), 7.10(s, 0.25H), 7.08-6.99(m, 3H), 6.96-6.84(m, 1.5H), 6.73(s, 0.25H), 4.23(s, 2H), 4.00(d, J=10.4Hz, 2H), 3.70(d, J=10.4Hz, 2H), 3.19(t, J=6.8Hz, 2H), 2.47(t , J=6.8Hz, 2H). MS-ESI calculated [M+H] + 617, found 617.
实施例12Example 12
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000050
Figure PCTCN2022111883-appb-000050
第一步first step
将化合物11-2(600mg,1.10mmol)溶于乙腈(5mL)中,加入碳酸钾(456mg,3.30mmol),碘化钾(548 mg,3.30mmol)和3-1(215mg,1.10mmol)。反应液在90℃搅拌反应12小时。向反应液加入水(20mL),用二氯甲烷(20mL x 2)萃取。合并有机相,用饱和食盐水洗涤(20mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(1:1石油醚/乙酸乙酯)得到化合物12-1。MS-ESI计算值[M+H] +659,实测值659。 Compound 11-2 (600 mg, 1.10 mmol) was dissolved in acetonitrile (5 mL), and potassium carbonate (456 mg, 3.30 mmol), potassium iodide (548 mg, 3.30 mmol) and 3-1 (215 mg, 1.10 mmol) were added. The reaction solution was stirred and reacted at 90° C. for 12 hours. Water (20 mL) was added to the reaction solution, and extracted with dichloromethane (20 mL x 2). The organic phases were combined, washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (1:1 petroleum ether/ethyl acetate) to obtain compound 12-1. MS-ESI calculated [M+H] + 659, found 659.
第二步second step
将化合物12-1(400mg,0.602mmol)和氢氧化钠(48.2mg,1.20mmol)加入到四氢呋喃(5mL)和水(1mL)中。反应液在氮气保护下60℃搅拌反应12小时。反应液减压浓缩。剩余物经高效液相色谱法(中性,碳酸氢铵体系)分离纯化得到化合物12。 1H NMR:(400MHz,CD 3OD)δ8.27-8.21(m,2H),8.03(d,J＝8.0Hz,1H),7.89-7.83(m,1H),7.63(d,J＝2.0Hz,1H),7.31(dd,J＝2.0,8.4Hz,1H),7.10(s,0.25H),7.07-7.00(m,3H),6.94-6.88(m,1.5H),6.73(s,0.25H),4.20(s,2H),3.86(d,J＝10.4Hz,2H),3.55(d,J＝10.4Hz,2H),2.91(t,J＝7.2Hz,2H),2.38(t,J＝7.2Hz,2H),1.78-1.71(m,2H)。MS-ESI计算值[M+H] +631,实测值631。 Compound 12-1 (400 mg, 0.602 mmol) and sodium hydroxide (48.2 mg, 1.20 mmol) were added to tetrahydrofuran (5 mL) and water (1 mL). The reaction solution was stirred and reacted at 60° C. for 12 hours under the protection of nitrogen. The reaction solution was concentrated under reduced pressure. The residue was separated and purified by high performance liquid chromatography (neutral, ammonium bicarbonate system) to obtain compound 12. 1 H NMR: (400MHz, CD 3 OD) δ8.27-8.21 (m, 2H), 8.03 (d, J = 8.0Hz, 1H), 7.89-7.83 (m, 1H), 7.63 (d, J = 2.0 Hz, 1H), 7.31(dd, J=2.0, 8.4Hz, 1H), 7.10(s, 0.25H), 7.07-7.00(m, 3H), 6.94-6.88(m, 1.5H), 6.73(s, 0.25H), 4.20(s, 2H), 3.86(d, J=10.4Hz, 2H), 3.55(d, J=10.4Hz, 2H), 2.91(t, J=7.2Hz, 2H), 2.38(t , J=7.2Hz, 2H), 1.78-1.71 (m, 2H). MS-ESI calculated [M+H] + 631, found 631.
实施例13Example 13
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000051
Figure PCTCN2022111883-appb-000051
第一步first step
将化合物11-2(600mg,1.10mmol)溶于乙腈(5mL)中,加入碳酸钾(456mg,3.30mmol),碘化钾(548mg,3.30mmol)和4-2(244mg,1.10mmol)。反应液在90℃搅拌反应12小时。向反应液加入水(20mL),用二氯甲烷(20mL x 2)萃取。合并有机相,用饱和食盐水(20mL x 1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(3:1石油醚/乙酸乙酯)得到化合物13-1。MS-ESI计算值[M+H] +671,实测值671。 Compound 11-2 (600 mg, 1.10 mmol) was dissolved in acetonitrile (5 mL), potassium carbonate (456 mg, 3.30 mmol), potassium iodide (548 mg, 3.30 mmol) and 4-2 (244 mg, 1.10 mmol) were added. The reaction solution was stirred and reacted at 90° C. for 12 hours. Water (20 mL) was added to the reaction solution, and extracted with dichloromethane (20 mL x 2). The organic phases were combined, washed with saturated brine (20 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (3:1 petroleum ether/ethyl acetate) to obtain compound 13-1. MS-ESI calculated [M+H] +671 , found 671.
第二步second step
将化合物13-1(400mg,0.580mmol)和氢氧化钠(46.4mg,1.16mmol)加入到四氢呋喃(5mL)和水(1 mL)中。反应液在氮气保护下60℃搅拌反应72小时。反应液减压浓缩。剩余物经高效液相色谱法(中性,碳酸氢铵体系)分离纯化得到化合物13。 1H NMR:(400MHz,CD 3OD)δ8.28-8.22(m,2H),8.05(d,J＝8.0Hz,1H),7.91-7.84(m,1H),7.63(d,J＝2.0Hz,1H),7.35(dd,J＝2.0,8.4Hz,1H),7.11-7.01(m,3H),6.98-6.86(m,2H),6.74(s,1H),4.27(s,2H),4.20(d,J＝10.4Hz,2H),3.93(d,J＝10.4Hz,2H),3.21(s,2H),1.23-1.14(m,2H),0.85-0.77(m,2H)。MS-ESI计算值[M+H] +643,实测值643。 Compound 13-1 (400 mg, 0.580 mmol) and sodium hydroxide (46.4 mg, 1.16 mmol) were added to tetrahydrofuran (5 mL) and water (1 mL). The reaction solution was stirred and reacted at 60° C. for 72 hours under the protection of nitrogen. The reaction solution was concentrated under reduced pressure. The residue was separated and purified by high performance liquid chromatography (neutral, ammonium bicarbonate system) to obtain compound 13. 1 H NMR: (400MHz, CD 3 OD) δ8.28-8.22 (m, 2H), 8.05 (d, J = 8.0Hz, 1H), 7.91-7.84 (m, 1H), 7.63 (d, J = 2.0 Hz,1H),7.35(dd,J=2.0,8.4Hz,1H),7.11-7.01(m,3H),6.98-6.86(m,2H),6.74(s,1H),4.27(s,2H) , 4.20 (d, J = 10.4Hz, 2H), 3.93 (d, J = 10.4Hz, 2H), 3.21 (s, 2H), 1.23-1.14 (m, 2H), 0.85-0.77 (m, 2H). MS-ESI calculated [M+H] + 643, found 643.
实施例14Example 14
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000052
Figure PCTCN2022111883-appb-000052
第一步first step
将化合物11-2(1.00g,1.84mmol)溶于1,2-二氯乙烷(10mL)中,在25℃下加入四乙氧基钛(838mg,3.67mmol)和5-2(478mg,3.67mmol)。反应液在25℃搅拌反应0.5小时,然后加入三乙酰氧基硼氢化钠(1.17g,5.51mmol),反应液在50℃搅拌反应2小时。向反应液加入水(50mL),用二氯甲烷(50mL x 2)萃取。合并有机相,用饱和食盐水(50mL x 1)洗涤,无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用硅胶柱色谱法分离纯化(3:1石油醚/乙酸乙酯)得到化合物14-1。MS-ESI计算值[M+H] +659,实测值659。 Compound 11-2 (1.00g, 1.84mmol) was dissolved in 1,2-dichloroethane (10mL), and tetraethoxytitanium (838mg, 3.67mmol) and 5-2 (478mg, 3.67 mmol). The reaction solution was stirred and reacted at 25°C for 0.5 hours, then sodium triacetoxyborohydride (1.17g, 5.51mmol) was added, and the reaction solution was stirred and reacted at 50°C for 2 hours. Water (50 mL) was added to the reaction solution, and extracted with dichloromethane (50 mL x 2). The organic phases were combined, washed with saturated brine (50 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (3:1 petroleum ether/ethyl acetate) to obtain compound 14-1. MS-ESI calculated [M+H] + 659, found 659.
第二步second step
将化合物14-1(2.00g,3.04mmol)和氢氧化钠(243mg,6.07mmol)加入到四氢呋喃(20mL)和水(4mL)中。反应液在氮气保护下60℃搅拌反应48小时。反应液减压浓缩。剩余物经高效液相色谱法(中性,碳酸氢铵体系)分离纯化得到化合物14。 1H NMR:(400MHz,CD 3OD)δ8.26-8.18(m,2H),8.03(d,J＝8.0Hz,1H),7.89-7.80(m,1H),7.65(d,J＝2.0Hz,1H),7.31(dd,J＝2.0,8.4Hz,1H),7.10(s,0.25H),7.07-6.96(m,3H),6.94-6.88(m,1.5H),6.73(s,0.25H),4.21(s,2H),3.86(d,J＝10.4Hz,2H),3.56(d,J＝10.4Hz,2H),2.93(s,2H),1.17(s,6H)。MS-ESI计算值[M+H] +645,实测值645。 Compound 14-1 (2.00 g, 3.04 mmol) and sodium hydroxide (243 mg, 6.07 mmol) were added to tetrahydrofuran (20 mL) and water (4 mL). The reaction solution was stirred and reacted at 60° C. for 48 hours under the protection of nitrogen. The reaction solution was concentrated under reduced pressure. The residue was separated and purified by high performance liquid chromatography (neutral, ammonium bicarbonate system) to obtain compound 14. 1 H NMR: (400MHz, CD 3 OD) δ8.26-8.18 (m, 2H), 8.03 (d, J = 8.0Hz, 1H), 7.89-7.80 (m, 1H), 7.65 (d, J = 2.0 Hz, 1H), 7.31(dd, J=2.0, 8.4Hz, 1H), 7.10(s, 0.25H), 7.07-6.96(m, 3H), 6.94-6.88(m, 1.5H), 6.73(s, 0.25H), 4.21(s, 2H), 3.86(d, J=10.4Hz, 2H), 3.56(d, J=10.4Hz, 2H), 2.93(s, 2H), 1.17(s, 6H). MS-ESI calculated [M+H] + 645, found 645.
实施例15Example 15
合成路线：synthetic route:
Figure PCTCN2022111883-appb-000053
Figure PCTCN2022111883-appb-000053
第一步first step
在2℃下,将亚硝酸钠(397mg,5.75mmol)的水(2.5mL)溶液加入化合物15-1(1g,5.23mmol)的浓盐酸(4.5mL)和水(24.5mL)的悬浊液中,在2℃下搅拌30分钟后于0℃向其中加入碘化钾(8.68g,52.3mmol)的水(5mL)溶液,反应液在15℃搅拌4小时。将反应液倒入冰中,用乙酸乙酯(50mL×2)萃取,合并有机相后依次用饱和碳酸氢钠水溶液(100mL×1)和水(100mL×1)洗涤,再用无水硫酸钠干燥,过滤,滤液减压浓缩后剩余物经过硅胶柱层析法(6/1石油醚/乙酸乙酯)分离纯化得到化合物15-2。 1H NMR(400MHz,CDCl 3)δ7.44-7.40(m,1H),7.31-7.29(m,1H),7.02-7.00(m,1H),3.96(s,3H)。 At 2°C, a solution of sodium nitrite (397mg, 5.75mmol) in water (2.5mL) was added to a suspension of compound 15-1 (1g, 5.23mmol) in concentrated hydrochloric acid (4.5mL) and water (24.5mL) After stirring at 2°C for 30 minutes, a solution of potassium iodide (8.68 g, 52.3 mmol) in water (5 mL) was added thereto at 0°C, and the reaction solution was stirred at 15°C for 4 hours. The reaction solution was poured into ice, extracted with ethyl acetate (50mL×2), and the combined organic phases were washed successively with saturated aqueous sodium bicarbonate (100mL×1) and water (100mL×1), and then washed with anhydrous sodium sulfate After drying and filtering, the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (6/1 petroleum ether/ethyl acetate) to obtain compound 15-2. 1 H NMR (400 MHz, CDCl 3 ) δ 7.44-7.40 (m, 1H), 7.31-7.29 (m, 1H), 7.02-7.00 (m, 1H), 3.96 (s, 3H).
第二步second step
将锌粉(486mg,7.43mmol)和1,2-二溴乙烷(99.0mg,0.524mmol)溶于N,N-二甲基甲酰胺(6.6mL)中,升温至70℃搅拌10分钟,降温至20℃后向其中滴加三甲基氯硅烷(57mg,0.524mmol),继续在20℃下搅拌50分钟,然后向其中加入化合物8-1(1.86g,6.56mmol)的N,N-二甲基甲酰胺(4mL)溶液,升温至40℃搅拌1小时,最后向其中快速加入化合物15-2(1.32g,4.37mmol),三(二亚苄基丙酮)二钯(80mg,87.4umol)和三(2-呋喃基)膦(41mg,0.175mmol)的N,N-二甲基甲酰胺(16mL)溶液,反应液在氮气保护下升温至60℃搅拌10小时。反应液通过硅藻土过滤,滤渣用乙酸乙酯(30mL)洗涤,滤液用饱和氯化铵水溶液(30mL×1)洗涤,有机相依次用水(30mL×3)和饱和食盐水(30mL×1)洗涤,再用无水硫酸钠干燥,过滤,滤液减压浓缩后剩余物经过硅胶柱层析法(4/1石油醚/乙酸乙酯)分离纯化得到化合物15-3。 1H NMR(400MHz,CDCl 3)δ7.37-7.33(m,1H),7.27-7.25(m,1H),7.15-7.13(m,1H),4.38-4.35(m,2H),4.23-4.13(m,3H),3.95(s,3H),1.51(s,9H)。MS-ESI计算值[M-56+H] +276,实测值276。 Dissolve zinc powder (486mg, 7.43mmol) and 1,2-dibromoethane (99.0mg, 0.524mmol) in N,N-dimethylformamide (6.6mL), heat up to 70°C and stir for 10 minutes, After cooling down to 20°C, trimethylchlorosilane (57mg, 0.524mmol) was added dropwise thereto, and stirring was continued at 20°C for 50 minutes, then compound 8-1 (1.86g, 6.56mmol) was added in N,N- Dimethylformamide (4mL) solution was heated to 40°C and stirred for 1 hour, and finally compound 15-2 (1.32g, 4.37mmol) and tris(dibenzylideneacetone)dipalladium (80mg, 87.4umol) were added rapidly therein ) and tris(2-furyl)phosphine (41mg, 0.175mmol) in N,N-dimethylformamide (16mL) solution, the reaction solution was heated to 60°C under nitrogen protection and stirred for 10 hours. The reaction solution was filtered through diatomaceous earth, the filter residue was washed with ethyl acetate (30mL), the filtrate was washed with saturated ammonium chloride aqueous solution (30mL×1), and the organic phase was sequentially washed with water (30mL×3) and saturated brine (30mL×1). After washing, drying with anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (4/1 petroleum ether/ethyl acetate) to obtain compound 15-3. 1 H NMR (400MHz, CDCl 3 )δ7.37-7.33(m,1H),7.27-7.25(m,1H),7.15-7.13(m,1H),4.38-4.35(m,2H),4.23-4.13 (m,3H), 3.95(s,3H), 1.51(s,9H). MS-ESI calculated value [M-56+H] + 276, found value 276.
第三步third step
参照实施例11第二步得到化合物15-4。MS-ESI计算值[M+H] +232,实测值232。 Referring to the second step of Example 11, compound 15-4 was obtained. MS-ESI calculated [M+H] + 232, found 232.
第四步the fourth step
将化合物5-3(350mg,0.606mmol),15-4(151mg,0.606mmol),碳酸铯(592mg,1.82mmol),4,5-双二苯基膦-9,9-二甲基氧杂蒽(35.1mg,0.0601mmol)和三(二亚苄基丙酮)二钯(27.8mg,0.0303mmol)加入到二氧六 环(5mL)中。反应液在氮气保护下90℃搅拌反应12小时。向反应液中加入水(10mL),用乙酸乙酯(10mL x 3)萃取。合并有机相,用饱和食盐水洗涤(10mL x 1),无水硫酸钠干燥,过滤,滤液减压浓缩。剩余物用薄层层析法分离纯化(3:1石油醚/乙酸乙酯)得到化合物15-5。MS-ESI计算值[M+H] +728,实测值728。 Compound 5-3 (350mg, 0.606mmol), 15-4 (151mg, 0.606mmol), cesium carbonate (592mg, 1.82mmol), 4,5-bisdiphenylphosphine-9,9-dimethyloxa Anthracene (35.1 mg, 0.0601 mmol) and tris(dibenzylideneacetone)dipalladium (27.8 mg, 0.0303 mmol) were added to dioxane (5 mL). The reaction solution was stirred and reacted at 90° C. for 12 hours under the protection of nitrogen. Water (10 mL) was added to the reaction solution, and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by thin layer chromatography (3:1 petroleum ether/ethyl acetate) to obtain compound 15-5. MS-ESI calculated value [M+H] + 728, found value 728.
第五步the fifth step
将化合物15-5(200mg,0.252mmol)和氢氧化钠(20.2mg,0.504mmol)加入到四氢呋喃(5mL)和水(1mL)中。反应液在氮气保护下60℃搅拌反应72小时。反应液减压浓缩。剩余物经高效液相色谱法(中性,碳酸氢铵体系)分离纯化得到化合物15。 1H NMR:(400MHz,CD 3OD)δ8.29-8.15(m,2H),7.99(d,J＝8.0Hz,1H),7.88-7.74(m,1H),7.48-7.34(m,1H),7.24(dd,J＝7.6,18.8Hz,2H),6.82(d,J＝8.0Hz,1H),6.57(s,1H),6.26(d,J＝7.6Hz,1H),4.38-4.35(m,1H),4.18-4.02(m,6H),3.79-3.72(m,2H),3.50-3.39(m,5H),2.91-2.85(m,2H),1.14(s,6H)。MS-ESI计算值[M+H] +714,实测值714。 Compound 15-5 (200 mg, 0.252 mmol) and sodium hydroxide (20.2 mg, 0.504 mmol) were added to tetrahydrofuran (5 mL) and water (1 mL). The reaction solution was stirred and reacted at 60° C. for 72 hours under the protection of nitrogen. The reaction solution was concentrated under reduced pressure. The residue was separated and purified by high performance liquid chromatography (neutral, ammonium bicarbonate system) to obtain compound 15. 1 H NMR: (400MHz, CD 3 OD) δ8.29-8.15(m, 2H), 7.99(d, J=8.0Hz, 1H), 7.88-7.74(m, 1H), 7.48-7.34(m, 1H ),7.24(dd,J=7.6,18.8Hz,2H),6.82(d,J=8.0Hz,1H),6.57(s,1H),6.26(d,J=7.6Hz,1H),4.38-4.35 (m,1H), 4.18-4.02(m,6H), 3.79-3.72(m,2H), 3.50-3.39(m,5H), 2.91-2.85(m,2H), 1.14(s,6H). MS-ESI calculated [M+H] + 714, found 714.
生物学评价：Biological Evaluation:
实验例1：本发明化合物对RORγ体外活性的测定Experimental example 1: Determination of the in vitro activity of the compound of the present invention on RORγ
1.1实验材料及仪器如表1所示。1.1 The experimental materials and instruments are shown in Table 1.
表1 实验材料及仪器Table 1 Experimental materials and instruments
实验材料及仪器Experimental Materials and Instruments 供应商supplier
hRORg-LBDhRORg-LBD HDBHDB
biotin-SRC1多肽biotin-SRC1 polypeptide 杭州中肽Hangzhou Zhongpei
SA-euSA-eu Perkin ElmerPerkin Elmer
SA-APCSA-APC Perkin ElmerPerkin Elmer
TrisTris SigmaSigma
KClKCl SigmaSigma
Na-EDTANa-EDTA InvitrogenInvitrogen
DTTDTT SigmaSigma
BSA(血清蛋白)BSA (serum protein) RocheRoche
Envision微孔板检测仪Envision Microplate Reader Perkin ElmerPerkin Elmer
1.2实验步骤1.2 Experimental steps
采用RORγ配体结合结构域(LBD)时间分辨荧光能量共振转移(TR-FRET)筛选本发明化合物对RORγ活性的调节。RORγ ligand-binding domain (LBD) time-resolved fluorescence energy transfer (TR-FRET) was used to screen the modulation of RORγ activity by the compounds of the present invention.
将待测化合物稀释在DMSO中，并进一步稀释在分析缓冲液(50mM Tris pH 7.0,50mM KCl,1mM Na-EDTA,0.1mM DTT,0.01％BSA)中(4倍稀释，10个浓度，最高浓度5000nM)，最终DMSO浓度1％。将hRORγ-LBD蛋白稀释在分析缓冲液中，得到在384孔板中15nM的hRORγ-LBD最终浓度。在分析缓冲液中制备biotin-SRC1多肽(Biotin-SPSSHSSLTERHKILHRLLQEGSP)储液，并加入到各孔中(终浓度 200nM)。将SA-eu(终浓度1nM)和SA-APC(终浓度50nM)的溶液也加入各孔中。The compound to be tested was diluted in DMSO and further diluted in assay buffer (50mM Tris pH 7.0, 50mM KCl, 1mM Na-EDTA, 0.1mM DTT, 0.01% BSA) (4-fold dilution, 10 concentrations, highest concentration 5000 nM), the final DMSO concentration was 1%. The hRORγ-LBD protein was diluted in assay buffer to give a final hRORγ-LBD concentration of 15 nM in 384-well plates. A stock solution of biotin-SRC1 polypeptide (Biotin-SPSSHSSSLTERHKILHRLLQEGSP) was prepared in assay buffer and added to each well (final concentration 200 nM). Solutions of SA-eu (1 nM final concentration) and SA-APC (50 nM final concentration) were also added to each well.
将最终分析混合物在4℃下孵育过夜，室温平衡1小时，1000rpm离心1分钟。在Envision微孔板检测仪上检测荧光读数，通过GraphPad Prism软件绘制发射波长665nM/615nM的荧光信号的比值与化合物浓度的对数曲线，计算化合物的50％有效浓度(EC 50)。最大应答(E max)为通过GraphPad Prism拟合确定的信号的上限峰值。 The final assay mixture was incubated overnight at 4°C, equilibrated at room temperature for 1 hour, and centrifuged at 1000 rpm for 1 minute. Fluorescence readings were detected on the Envision microplate detector, and the logarithmic curve of the ratio of the fluorescence signal at emission wavelength 665nM/615nM to the compound concentration was drawn by GraphPad Prism software, and the 50% effective concentration (EC 50 ) of the compound was calculated. The maximum response (E max ) was the upper peak peak of the signal determined by GraphPad Prism fit.
1.3实验结果如表2所示。1.3 The experimental results are shown in Table 2.
表2 本发明化合物对RORγ体外活性的测定结果Table 2 The assay results of the compounds of the present invention to the in vitro activity of RORγ
化合物编号Compound number EC 50(nM) EC50 (nM) E max Emax
化合物1Compound 1 66.366.3 77.5％77.5%
化合物2Compound 2 65.765.7 82.0％82.0%
化合物8Compound 8 85.9785.97 102.27102.27
化合物9Compound 9 48.748.7 64.4％64.4%
化合物10Compound 10 25.3425.34 108.94％108.94%
化合物11Compound 11 7.0957.095 118.75％118.75%
化合物12Compound 12 3.5433.543 129.13％129.13%
化合物13Compound 13 12.5912.59 106.62％106.62%
化合物14Compound 14 34.8434.84 122.50％122.50%
化合物15Compound 15 69.1669.16 120.70％120.70%
结论：本发明化合物对RORγ体外活性具有明显的激动作用。Conclusion: the compound of the present invention has obvious agonistic effect on the in vitro activity of RORγ.
实验例2：本发明化合物促进CD4 +细胞分化为Th17细胞的能力测定 Experimental example 2: Determination of the ability of the compounds of the present invention to promote the differentiation of CD4 + cells into Th17 cells
2.1实验材料及仪器2.1 Experimental materials and instruments
1.C57BL/6小鼠脾脏1. C57BL/6 mouse spleen
2. 70μm滤网(BD)2. 70μm filter (BD)
3.红细胞裂解液(Absin)3. Red blood cell lysate (Absin)
4.Mouse CD4+T cell isolation kit(小鼠CD4+T细胞分选试剂盒)(Stemcell)4. Mouse CD4+T cell isolation kit (Mouse CD4+T cell isolation kit) (Stemcell)
5.CD3抗体(BD)5. CD3 antibody (BD)
6.CD28抗体(BD)6. CD28 antibody (BD)
7. 96孔U底板(Corning)7. 96-well U-base plate (Corning)
8.DPBS(Corning)8. DPBS (Corning)
9.TGFβ(R&D)9. TGFβ (R&D)
10.IL-6(R&D)10. IL-6 (R&D)
11.IL-1β(BioLegend)11. IL-1β (BioLegend)
12.IL-23(BioLegend)12. IL-23 (BioLegend)
13.RPMI 1640培养液(Gibco)13. RPMI 1640 culture medium (Gibco)
14.青/链霉素(HyClone)14. Penicillin/Streptomycin (HyClone)
15.胎牛血清(HyClone)15. Fetal bovine serum (HyClone)
16.非必需氨基酸(Gibco)16. Non-essential amino acids (Gibco)
17.β-巯基乙醇(Sigma)17. β-Mercaptoethanol (Sigma)
18.IFNγ抗体(BD)18. IFNγ antibody (BD)
19.IL-4抗体(BD)19. IL-4 antibody (BD)
20.PMA(sigma)20. PMA (sigma)
21.Ionomycin(离子霉素)(Invitrogen)21. Ionomycin (Ionomycin) (Invitrogen)
22.高尔基体抑制剂(BD)22. Golgi inhibitors (BD)
23.Staining buffer(染色缓冲液)(Biolegend)23. Staining buffer (Biolegend)
24.Fixation buffer(固定缓冲液)(Biolegend)24. Fixation buffer (Biolegend)
25.Permeabilization buffer(透化缓冲液)(Biolegend)25. Permeabilization buffer (Biolegend)
26.LIVE/DEAD stain kit(染色试剂盒)(Invitrogen)26. LIVE/DEAD stain kit (staining kit) (Invitrogen)
27.CD4抗体(Biolegend)27. CD4 antibody (Biolegend)
28.IL-17A抗体(Biolegend)28. IL-17A antibody (Biolegend)
29.细胞计数仪(Beckman)29. Cell counter (Beckman)
30.离心机(Eppendorf)30. Centrifuge (Eppendorf)
31.流式细胞仪(BD)31. Flow Cytometry (BD)
32.CO 2培养箱(Thermo) 32. CO2 incubator (Thermo)
2.2实验步骤2.2 Experimental steps
将CD3抗体于DPBS中稀释至5μg/mL，加入96孔U底板中，每孔50μL液体，于4℃包被过夜。The CD3 antibody was diluted to 5 μg/mL in DPBS, added to a 96-well U-bottom plate, 50 μL of liquid per well, and coated overnight at 4°C.
在培养液(RPMI 1640+10％胎牛血清+1％青链霉素+1％非必需氨基酸+0.05mMβ-巯基乙醇)中研磨C57BL/6小鼠脾脏，过70μm滤网制备单细胞悬液，300g离心3min。加入红细胞裂解液室温裂解3min。使用小鼠CD4 +T细胞分离试剂盒分离CD4 +细胞。将前一天包被的96孔U底板取出，吸去包被的液体，并用DPBS洗两次，再将上面得到的CD4 +细胞以5*10 5/mL的密度接种至包被过的孔中，每孔200μL细胞悬液；再加入CD28抗体(3μg/mL)、TGFβ(3ng/mL)、IL-6(30ng/mL)、IL-23(10ng/mL)、IL-1β(10ng/mL)、IFNγ抗体(10μg/mL)和IL-4抗体(10μg/mL)；再在孔中加入本发明化合物，于37℃， 5％CO 2下培养3天。 Grind C57BL/6 mouse spleen in culture medium (RPMI 1640 + 10% fetal bovine serum + 1% penicillin + 1% non-essential amino acid + 0.05mM β-mercaptoethanol), and pass through a 70μm filter to prepare a single cell suspension , centrifuge at 300g for 3min. Add erythrocyte lysate to lyse at room temperature for 3 min. CD4 + cells were isolated using the mouse CD4 + T cell isolation kit. Take out the 96-well U-bottom plate coated the day before, suck off the coated liquid, wash twice with DPBS, and inoculate the CD4 + cells obtained above at a density of 5*10 5 /mL into the coated wells , 200μL cell suspension per well; then add CD28 antibody (3μg/mL), TGFβ (3ng/mL), IL-6 (30ng/mL), IL-23 (10ng/mL), IL-1β (10ng/mL ), IFNγ antibody (10 μg/mL) and IL-4 antibody (10 μg/mL); then the compound of the present invention was added to the well, and cultured at 37° C. under 5% CO 2 for 3 days.
在每个孔中加入500ng/mL的PMA和离子霉素，以及高尔基体抑制剂(1:1000)，37℃，5％CO 2下刺激4小时。 Add 500ng/mL of PMA and ionomycin, and Golgi inhibitor (1:1000) to each well, and stimulate for 4 hours at 37°C and 5% CO 2 .
刺激结束后将U底板300g离心3min，倒掉上清，用染色缓冲液洗两次。将CD4抗体和LIVE/DEAD染液分别按照1:200和1:1000于染色缓冲液中稀释，每孔加入50μL染液，4℃染色30min，之后用染色缓冲液洗涤细胞两次。每孔加入100μL固定缓冲液，室温固定20min，用透化缓冲液洗两次。将IL-17A抗体按照1：200于透化缓冲液中稀释，每孔加入50μL染液，室温染色30min，之后用染色缓冲液洗两次。最后用150μL染色缓冲液重悬细胞，使用流式细胞仪检测Th17细胞比例。After the stimulation, the U-bottom plate was centrifuged at 300 g for 3 min, the supernatant was discarded, and washed twice with staining buffer. Dilute CD4 antibody and LIVE/DEAD staining solution in staining buffer at 1:200 and 1:1000, respectively, add 50 μL of staining solution to each well, stain at 4°C for 30 min, and then wash cells twice with staining buffer. Add 100 μL of fixation buffer to each well, fix at room temperature for 20 min, and wash twice with permeabilization buffer. The IL-17A antibody was diluted 1:200 in permeabilization buffer, 50 μL of dye solution was added to each well, stained at room temperature for 30 min, and then washed twice with staining buffer. Finally, the cells were resuspended with 150 μL staining buffer, and the proportion of Th17 cells was detected by flow cytometry.
2.3实验结果如表3所示。2.3 The experimental results are shown in Table 3.
表3 本发明化合物促进CD4 +细胞分化为Th17细胞的能力测定结果 Table 3 The results of the determination of the ability of the compounds of the present invention to promote the differentiation of CD4 + cells into Th17 cells
化合物编号Compound number Th17细胞比例Th17 cell ratio
DMSODMSO 26.5％26.5%
化合物1Compound 1 47.3％47.3%
化合物5Compound 5 52.9％52.9%
化合物8Compound 8 41.7％41.7%
化合物测试浓度：1μM。Compound test concentration: 1 μM.
结论：本发明化合物可以明显地促进CD4 +细胞分化为Th17细胞。 Conclusion: the compounds of the present invention can obviously promote the differentiation of CD4 + cells into Th17 cells.
实验例3：本发明化合物药代动力学评价Experimental example 3: Pharmacokinetic evaluation of the compound of the present invention
3.1实验目的：测试化合物在Balb/c小鼠体内的药代动力学3.1 Purpose of the experiment: to test the pharmacokinetics of compounds in Balb/c mice
3.2实验材料：3.2 Experimental materials:
Balb/c小鼠(雌性,7～9周龄,上海斯莱克)Balb/c mice (female, 7-9 weeks old, Shanghai Slack)
3.3实验操作：3.3 Experimental operation:
以标准方案测试化合物静脉注射及口服给药后的啮齿类动物药代特征,实验中给予小鼠单次静脉注射(IV)及口服给药(PO)。静注溶媒为5％二甲基亚砜、30％的PEG400、65％的10％羟丙基β环糊精配成的混合溶媒。口服溶媒为0.5％羟丙甲纤维素和0.2％吐温配成的混合溶媒。该项目使用四只雌性Balb/c小鼠,两只小鼠进行静脉注射给药,给药剂量为0.5mg/kg,收集0h(给药前)和给药后0.0833,0.25,0.5,1,2,4,8,24h的血浆样品,另外两只小鼠口服灌胃给药,给药剂量为1mg/kg,收集0h(给药前)和给药后0.25,0.5,1,2,4,8,24h的血浆样品,收集24小时内的全血样品,3000g离心15分钟,分离上清得血浆样品,加入4倍体积含内标的乙腈溶液沉淀蛋白,离心取上清液加入等倍体积的水再离心取上清进样,以LC-MS/MS分析方法定量分析血药浓度,并计算药代参数,如达峰浓度(C max),清除率(CL),半衰期(T 1/2),组织分布(Vdss),药时曲 线下面积(AUC 0-last),生物利用度(F)。 The pharmacokinetic characteristics of the compounds were tested in rodents after intravenous injection and oral administration according to the standard protocol. In the experiment, the mice were given a single intravenous injection (IV) and oral administration (PO). The solvent for intravenous injection is a mixed solvent made up of 5% dimethyl sulfoxide, 30% PEG400, and 65% 10% hydroxypropyl β-cyclodextrin. The oral vehicle is a mixed vehicle made of 0.5% hypromellose and 0.2% Tween. The project used four female Balb/c mice, two mice were administered intravenously, the dose was 0.5mg/kg, and the collection 0h (before administration) and after administration were 0.0833, 0.25, 0.5, 1, Plasma samples at 2, 4, 8, and 24 hours were administered orally to the other two mice at a dose of 1 mg/kg, collected at 0 h (before administration) and at 0.25, 0.5, 1, 2, and 4 hours after administration , 8, 24h plasma samples, collect whole blood samples within 24 hours, centrifuge at 3000g for 15 minutes, separate supernatant to obtain plasma samples, add 4 times volume of acetonitrile solution containing internal standard to precipitate protein, centrifuge to take supernatant and add equal volume The water was then centrifuged to take the supernatant sample, and the blood drug concentration was quantitatively analyzed by LC-MS/MS analysis method, and the pharmacokinetic parameters were calculated, such as peak concentration (C max ), clearance rate (CL), half-life (T 1 / 2 ), tissue distribution (Vdss), area under the drug-time curve (AUC 0-last ), bioavailability (F).
3.4实验结果如表4所示。3.4 The experimental results are shown in Table 4.
表4 本发明化合物的药代动力学测试结果The pharmacokinetic test result of table 4 compound of the present invention
Figure PCTCN2022111883-appb-000054
Figure PCTCN2022111883-appb-000054
结论：本发明化合物具有良好的药代动力学性质,包括良好的口服生物利用度,口服暴露量,半衰期和清除率等。Conclusion: The compounds of this invention have good pharmacokinetic properties, including good oral bioavailability, oral exposure, half-life and clearance rate.
实验例4：本发明化合物对MC38小鼠结肠癌移植瘤模型的体内药效学研究Experimental Example 4: In vivo pharmacodynamic study of the compound of the present invention on MC38 mouse colon cancer xenograft model
4.1实验目的：4.1 Purpose of the experiment:
本实验的目的是研究本发明化合物对MC38小鼠结肠癌移植瘤模型体内药效进行评估。The purpose of this experiment is to study the evaluation of the compound of the present invention on the MC38 mouse colon cancer xenograft tumor model in vivo.
4.2实验动物：4.2 Experimental animals:
种属：小鼠Species: mouse
品系：C57BL/6小鼠Strain: C57BL/6 mice
周龄及体重：7周龄,体重18-23克Week age and weight: 7 weeks old, weighing 18-23 grams
性别：雌性Gender: female
供应商：上海斯莱克实验动物有限公司Supplier: Shanghai Slack Experimental Animal Co., Ltd.
4.3实验方法与步骤4.3 Experimental methods and steps
4.3.1细胞培养4.3.1 Cell culture
名称：MC38(小鼠结肠癌细胞)Name: MC38 (Mouse Colon Cancer Cell)
来源：和元生物技术(上海)有限公司。由辉源生物科技(上海)有限公司保种维持传代。Source: Heyuan Biotechnology (Shanghai) Co., Ltd. The species was maintained and passed on by Huiyuan Biotechnology (Shanghai) Co., Ltd.
细胞培养：培养液为含有10％胎牛血清的1640培养基，培养条件为37℃,5％二氧化碳。传代比例为1:2～1:3,每周传代2～3次。Cell culture: the culture medium is 1640 medium containing 10% fetal bovine serum, and the culture condition is 37°C, 5% carbon dioxide. The subculture ratio was 1:2～1:3, and subcultured 2～3 times a week.
4.3.2肿瘤细胞接种4.3.2 Tumor cell inoculation
将0.1mL(2×10 5个)细胞皮下接种于每只小鼠的右后背。同日将动物根据体重随机分组。 0.1 mL (2×10 5 ) cells were inoculated subcutaneously on the right back of each mouse. On the same day, animals were randomized into groups based on body weight.
4.3.3受试物的配制4.3.3 Preparation of test substances
实验用溶媒为5％DMSO/95％(20％羟丙基β环糊精)。受试物用溶媒溶解,配制成一定浓度均一溶液,于4℃保存。The experimental vehicle was 5% DMSO/95% (20% hydroxypropyl beta cyclodextrin). The test substance was dissolved in a solvent, prepared into a uniform solution with a certain concentration, and stored at 4°C.
4.3.4肿瘤测量和实验指标4.3.4 Tumor measurements and experimental indicators
实验指标是考察肿瘤生长是否被抑制、延缓或治愈。每周两次用游标卡尺测量肿瘤直径。肿瘤体积的计算公式为：V＝0.5a×b 2,a和b分别表示肿瘤的长径和短径。 The experimental index is to investigate whether tumor growth is inhibited, delayed or cured. Tumor diameters were measured twice a week with vernier calipers. The calculation formula of tumor volume is: V=0.5a×b 2 , where a and b represent the long diameter and short diameter of the tumor respectively.
化合物的抑瘤疗效用相对肿瘤增殖率T/C(％)评价。相对肿瘤增殖率T/C(％)：计算公式如下：T/C％＝T RTV/C RTV×100％(T RTV：治疗组RTV；C RTV：阴性对照组RTV)。根据肿瘤测量的结果计算出相对肿瘤体积(relative tumor volume,RTV)，计算公式为RTV＝V t/V 0，其中V 0是分组给药时(即d 0)测量所得平均肿瘤体积，V t为某一次测量时的平均肿瘤体积，T RTV与C RTV取同一天数据。 The antitumor efficacy of compounds was evaluated by relative tumor proliferation rate T/C (%). Relative tumor proliferation rate T/C (%): the calculation formula is as follows: T/C%=T RTV /C RTV ×100% (T RTV : RTV of the treatment group; C RTV : RTV of the negative control group). Calculate the relative tumor volume (RTV) according to the results of tumor measurement, and the calculation formula is RTV=V t /V 0 , where V 0 is the average tumor volume measured during group administration (that is, d 0 ), V t is the average tumor volume at a certain measurement, and T RTV and C RTV take the data of the same day.
4.4实验结果如表5所示。4.4 The experimental results are shown in Table 5.
表5 本发明化合物对MC38小鼠结肠癌移植瘤模型的抑瘤药效评价(基于给药后第28天肿瘤体积计算得出)Table 5 Evaluation of the antitumor efficacy of the compounds of the present invention on the MC38 mouse colon cancer xenograft model (calculated based on the tumor volume on the 28th day after administration)
Figure PCTCN2022111883-appb-000055
Figure PCTCN2022111883-appb-000055
PD-1单抗来源：BioXcell。PD-1单抗于分组后第7天开始给药，化合物1于分组当天开始给药。Source of PD-1 monoclonal antibody: BioXcell. PD-1 monoclonal antibody was administered on the 7th day after grouping, and compound 1 was administered on the day of grouping.
结论：本发明化合物与PD-1单抗联用对MC38小鼠结肠癌移植瘤模型具有优异的抑瘤效果。Conclusion: The combination of the compound of the present invention and PD-1 monoclonal antibody has an excellent tumor-inhibiting effect on the transplanted tumor model of MC38 mouse colon cancer.

Claims (14)

  1. 式(Ⅰ)化合物或其药学上可接受的盐，A compound of formula (I) or a pharmaceutically acceptable salt thereof,
    Figure PCTCN2022111883-appb-100001
    Figure PCTCN2022111883-appb-100001
    其中，in,
    R 11、R 12、R 13、R 14和R 15分别独立地选自H、F、Cl、Br、I、-OH、-NH 2、-CN、C 1-3烷基、C 1-3烷氧基和C 1-3烷氨基，其中所述C 1-3烷基、C 1-3烷氧基和C 1-3烷氨基分别独立地任选被1、2或3个R a取代； R 11 , R 12 , R 13 , R 14 and R 15 are independently selected from H, F, Cl, Br, I, -OH, -NH 2 , -CN, C 1-3 alkyl, C 1-3 Alkoxy and C 1-3 alkylamino, wherein said C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino are independently optionally substituted by 1, 2 or 3 R a ;
    R 2选自C 1-6烷基和‐C 1-3烷基-C 3-4环烷基，所述C 1-6烷基和‐C 1-3烷基-C 3-4环烷基分别独立地任选被1、2或3个R b取代； R 2 is selected from C 1-6 alkyl and -C 1-3 alkyl-C 3-4 cycloalkyl, said C 1-6 alkyl and -C 1-3 alkyl-C 3-4 cycloalkane The groups are independently optionally substituted by 1, 2 or 3 R b ;
    L 1选自单键、-C 2-4烯基-和-4-5元杂环烷基-，所述-C 2-4烯基-和-4-5元杂环烷基-分别独立地任选被1、2或3个R c取代； L is selected from a single bond, -C 2-4 alkenyl- and -4-5 membered heterocycloalkyl-, and the -C 2-4 alkenyl- and -4-5 membered heterocycloalkyl- are independently optionally substituted by 1, 2 or 3 R c ;
    R a分别独立地选自F、Cl、Br、I、-OH、-NH 2和C 1-3烷基，其中所述C 1-3烷基任选被1、2或3个R取代； R a is independently selected from F, Cl, Br, I, -OH, -NH 2 and C 1-3 alkyl, wherein the C 1-3 alkyl is optionally substituted by 1, 2 or 3 R;
    R b分别独立地选自F、Cl、Br、I、-OH、-NH 2、-CN、-COOH和C 1-3烷基； R b are independently selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and C 1-3 alkyl;
    R c分别独立地选自F、Cl、Br、I、-OH、-NH 2、-CN、-COOH和C 1-3烷基； R c are independently selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and C 1-3 alkyl;
    R分别独立地选自F、Cl、Br、I、-OH和-NH 2； R is independently selected from F, Cl, Br, I, -OH and -NH 2 ;
    所述-4-5元杂环烷基-中的“杂”表示1、2、3或4个分别独立选自-O-、-NH-、-S-和N的杂原子或杂原子团。"Hetero" in the -4-5 membered heterocycloalkyl- means 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from -O-, -NH-, -S- and N, respectively.
  2. 根据权利要求1所述化合物或其药学上可接受的盐，其中，R a选自F、Cl、Br、I、-OH、-NH 2和-CH 3。 The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R a is selected from F, Cl, Br, I, -OH, -NH 2 and -CH 3 .
  3. 根据权利要求1所述化合物或其药学上可接受的盐，其中，R b选自F、Cl、Br、I、-OH、-NH 2、-CN、-COOH和-CH 3。 The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R b is selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and -CH 3 .
  4. 根据权利要求1所述化合物或其药学上可接受的盐，其中，R c选自F、Cl、Br、I、-OH、-NH 2、-CN、-COOH和-CH 3。 The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R c is selected from F, Cl, Br, I, -OH, -NH 2 , -CN, -COOH and -CH 3 .
  5. 根据权利要求1所述化合物或其药学上可接受的盐，其中，R 11、R 12、R 13、R 14和R 15分别独立地选自H、F、Cl、Br、I、-OH、-NH 2、-CN、-CH 3、-O-CH 3和-NH-CH 3，其中所述-CH 3、-O-CH 3和-NH-CH 3分别独立地任选被1、2或3个R a取代。 The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 11 , R 12 , R 13 , R 14 and R 15 are independently selected from H, F, Cl, Br, I, -OH, -NH 2 , -CN, -CH 3 , -O-CH 3 and -NH-CH 3 , wherein the -CH 3 , -O-CH 3 and -NH-CH 3 are independently optionally replaced by 1, 2 or 3 R a substitutions.
  6. 根据权利要求5所述化合物或其药学上可接受的盐，其中，R 11、R 12、R 13、R 14和R 15选自H、F、Cl、Br、I、-OH、-NH 2、-CN、-CH 3、-CF 3、-O-CH 3、-O-CH 2F、-O-CHF 2、-O-CF 3和
    Figure PCTCN2022111883-appb-100002
    The compound or a pharmaceutically acceptable salt thereof according to claim 5, wherein R 11 , R 12 , R 13 , R 14 and R 15 are selected from H, F, Cl, Br, I, -OH, -NH 2 , -CN, -CH 3 , -CF 3 , -O-CH 3 , -O-CH 2 F, -O-CHF 2 , -O-CF 3 and
    Figure PCTCN2022111883-appb-100002
  7. 根据权利要求1所述化合物或其药学上可接受的盐，其中，R 2选自C 1-4烷基和‐C 1-3烷基-环丙基，其中所述C 1-4烷基和‐C 1-3烷基-环丙基分别独立地任选被1、2或3个R b取代。 The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R is selected from C 1-4 alkyl and -C 1-3 alkyl-cyclopropyl, wherein the C 1-4 alkyl and -C 1-3 alkyl-cyclopropyl are each independently optionally substituted by 1, 2 or 3 R b .
  8. 根据权利要求7所述化合物或其药学上可接受的盐，其中，R 2选自-CH 3、-CH 2-CH 3、-CH 2-CH 2-CH 3、
    Figure PCTCN2022111883-appb-100003
    其中所述-CH 3、-CH 2-CH 3、-CH 2-CH 2-CH 3、
    Figure PCTCN2022111883-appb-100004
    分别独立地任选被1、2或3个R b取代。     The compound or a pharmaceutically acceptable salt thereof according to claim 7, wherein R 2 is selected from -CH 3 , -CH 2 -CH 3 , -CH 2 -CH 2 -CH 3 ,
    Figure PCTCN2022111883-appb-100003
    Wherein -CH 3 , -CH 2 -CH 3 , -CH 2 -CH 2 -CH 3 ,
    Figure PCTCN2022111883-appb-100004
    are each independently optionally substituted by 1, 2 or 3 R b .
  9. 根据权利要求8所述化合物或其药学上可接受的盐，其中，R 2选自
    Figure PCTCN2022111883-appb-100005
    Figure PCTCN2022111883-appb-100006
    The compound or a pharmaceutically acceptable salt thereof according to claim 8, wherein R is selected from
    Figure PCTCN2022111883-appb-100005
    Figure PCTCN2022111883-appb-100006
  10. 根据权利要求1所述化合物或其药学上可接受的盐，其中，L 1选自单键、-C 2-3烯基-和-吖丁啶基‐，其中所述-C 2-3烯基-和-吖丁啶基‐分别独立地任选被1、2或3个R c取代。 The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein L is selected from single bonds, -C 2-3 alkenyl- and -azetidinyl-, wherein the -C 2-3 alkenyl- and -azetidinyl- each independently optionally substituted by 1, 2 or 3 R c .
  11. 根据权利要求10所述化合物或其药学上可接受的盐，其中，L 1选自单键、-CH＝CH-和
    Figure PCTCN2022111883-appb-100007
    其中所述-CH＝CH-和
    Figure PCTCN2022111883-appb-100008
    分别独立地任选被1、2或3个R c取代。     The compound or a pharmaceutically acceptable salt thereof according to claim 10, wherein L is selected from a single bond, -CH=CH- and
    Figure PCTCN2022111883-appb-100007
    Wherein said -CH=CH- and
    Figure PCTCN2022111883-appb-100008
    are each independently optionally substituted by 1, 2 or 3 R c .
  12. 根据权利要求11所述化合物或其药学上可接受的盐，其中，L 1选自单键、-CH＝CH-、-C(CH 3)＝CH-和
    Figure PCTCN2022111883-appb-100009
    The compound according to claim 11 or a pharmaceutically acceptable salt thereof, wherein L 1 is selected from a single bond, -CH=CH-, -C(CH 3 )=CH- and
    Figure PCTCN2022111883-appb-100009
  13. 根据权利要求1所述化合物或其药学上可接受的盐，其中，结构单元
    Figure PCTCN2022111883-appb-100010
    选自
    Figure PCTCN2022111883-appb-100011
    Figure PCTCN2022111883-appb-100012
    The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit
    Figure PCTCN2022111883-appb-100010
    selected from
    Figure PCTCN2022111883-appb-100011
    Figure PCTCN2022111883-appb-100012
  14. 下列所示化合物或其药学上可接受的盐，Compounds shown below or pharmaceutically acceptable salts thereof,
    Figure PCTCN2022111883-appb-100013
    Figure PCTCN2022111883-appb-100013
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