WO2023078411A1 - Azaspiro compound - Google Patents

Azaspiro compound Download PDF

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WO2023078411A1
WO2023078411A1 PCT/CN2022/129958 CN2022129958W WO2023078411A1 WO 2023078411 A1 WO2023078411 A1 WO 2023078411A1 CN 2022129958 W CN2022129958 W CN 2022129958W WO 2023078411 A1 WO2023078411 A1 WO 2023078411A1
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compound
mmol
reaction
mobile phase
dissolved
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PCT/CN2022/129958
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French (fr)
Chinese (zh)
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吴凌云
王才林
徐雄彬
赵乐乐
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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    • 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
    • A61K31/66—Phosphorus compounds
    • A61K31/675—Phosphorus compounds having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6558—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system

Definitions

  • the present invention relates to a class of azaspiro compounds, in particular to the application of compounds represented by formula (II-1) or pharmaceutically acceptable salts thereof in the preparation of drugs for treating related diseases.
  • LRRK2 kinase are increasingly proven to be the fundamental factors in inducing neurodegenerative diseases, characterized by selective degeneration and cell death of dopaminergic neurons in the substantia nigra. Affects 1% of the population over the age of 65, of which genetic patients account for 5-10% of the affected population. Early in the disease, the most obvious symptoms are shaking, slowness of movement and difficulty walking. Cognitive and behavioral problems also develop later, and dementia usually occurs later.
  • LRRK2 leucine-rich repeat kinase 2
  • NM_198578.2 The NCBI participating sequence for human LRRK2 mRNA.
  • Evidence shows that LRRK2 phosphorylates ⁇ -synuclein at serine-129 and that this phosphorylated form constitutes an important part of Lewy bodies.
  • SNPs in the functional domains of LRRK2 have been shown to cause common and sporadic neurodegenerative diseases.
  • LRRK2 mutations have identified more than 20 LRRK2 mutations in families with late-onset neurodegenerative disease.
  • the G2019S mutation co-segregates with autosomal dominant and it is responsible for approximately 6% of familial cases and 3% of sporadic cases in Europe.
  • the G2019S mutation occurs in a highly conserved kinase domain, so the G2019S mutation may have an effect on kinase activity.
  • an amino acid substitution at another residue, R1441 has also been associated with neurodegenerative diseases and was shown to increase LRRK2 kinase activity.
  • the present invention aims to invent a compound that can highly inhibit LRRK2 kinase, so as to further invent a drug that can well treat neurodegenerative diseases.
  • J.Med.Chem.2012,55,9416-9433 discloses compound GNE-7915, which belongs to LRRK2 kinase inhibitor, and its structural formula is as follows:
  • the present invention provides compounds represented by formulas (III-1), (II-2), (II-3), (II-4) and (III-2) or pharmaceutically acceptable salts thereof:
  • X 1 is CH, X 2 is CA; or, X 1 is CA, X 2 is CH;
  • Y is -C(R 9 R 10 )- or -O-;
  • Z 1 is CH, Z 2 is N; or, Z 1 is N, Z 2 is CH;
  • R 1 and R 2 are independently selected from H, F, Cl, Br, I, CH 3 and CF 3 ;
  • R 3 and R 4 are independently selected from H and C 1-3 alkyl, and the C 1-3 alkyl is optionally substituted by 1, 2 or 3 halogens;
  • R 5 and R 6 are independently selected from H, F, CH 3 and CF 3 ;
  • R 7 is selected from CH 3 and CHF 2 ;
  • R 9 and R 10 are independently selected from H, F, Cl, Br, I, C 1-3 alkyl and C 1-3 alkoxy, the C 1-3 alkyl and C 1-3 alkoxy Each group is independently optionally substituted by 1, 2 or 3 halogens;
  • n, q, t are independently selected from 0, 1 and 2, respectively.
  • the above-mentioned compounds are selected from compounds shown in formulas (II-1), (II-2), (II-3) and (II-4):
  • X 1 is CH, X 2 is CA; or, X 1 is CA, X 2 is CH;
  • Y is -C(R 9 R 10 )- or -O-;
  • Z 1 is CH, Z 2 is N; or, Z 1 is N, Z 2 is CH;
  • R 1 and R 2 are independently selected from H, F, Cl, Br, I, CH 3 and CF 3 ;
  • R 3 and R 4 are independently selected from H and C 1-3 alkyl, and the C 1-3 alkyl is optionally substituted by 1, 2 or 3 halogens;
  • R 5 and R 6 are independently selected from H and F;
  • R 7 is selected from CH 3 and CHF 2 ;
  • R 9 and R 10 are independently selected from H, F, Cl, Br, I, C 1-3 alkyl and C 1-3 alkoxy, the C 1-3 alkyl and C 1-3 alkoxy Each group is independently optionally substituted by 1, 2 or 3 halogens;
  • n, q, t are independently selected from 0, 1 and 2, respectively.
  • the above compounds are selected from:
  • R 1 , R 2 , R 3 , R 6 and R 7 are as defined in the present invention.
  • Y, R 1 , R 2 , R 3 , R 4 , m, n, q and t are as defined in the present invention.
  • Y is -C(R 9 R 10 )- or -O-;
  • R 1 and R 2 are independently selected from H, F, Cl, Br, I, CH 3 and CF 3 ;
  • R 3 and R 4 are independently selected from H and C 1-3 alkyl, and the C 1-3 alkyl is optionally substituted by 1, 2 or 3 halogens;
  • R 9 and R 10 are independently selected from H, F, Cl, Br, I, C 1-3 alkyl and C 1-3 alkoxy, the C 1-3 alkyl and C 1-3 alkoxy Each group is independently optionally substituted by 1, 2 or 3 halogens;
  • n, q, t are independently selected from 0, 1 and 2, respectively.
  • R 1 is selected from H and F, and other variables are as defined in the present invention.
  • R 2 is selected from Cl and F, and other variables are as defined in the present invention.
  • R 3 and R 4 are independently selected from CH 3 , and other variables are as defined in the present invention.
  • R 9 and R 10 are independently selected from H, F, CH 3 , -OCHF 2 and -OCH 3 , and other variables are as defined in the present invention.
  • R 9 and R 10 are independently selected from H, F and -OCH 3 , and other variables are as defined in the present invention.
  • the above-mentioned Y is -CHF-, -CF 2 -, -CH 2 -, -CH(OCH 3 )-, -CH(OCHF 2 )- and -O-, other variables are as in the present invention defined.
  • the present invention also provides a compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from
  • the present invention also provides the application of the above compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating Parkinson's disease.
  • the compound of the present invention has remarkable kinase inhibition, cell activity, membrane permeability and solubility for LRRK2, and has excellent pharmacokinetic and pharmacodynamic properties, such as better brain entry properties.
  • 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 and the like , 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
  • 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 terms "effective amount” or “therapeutically effective amount” refer to a non-toxic amount that achieves the desired effect. The determination of the effective amount varies from person to person, depending on the age and general condition of the recipient, and also depends on the specific active substance. The appropriate effective amount in each case can be determined by those skilled in the art according to routine experiments.
  • 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 refers to the replacement of any one or more hydrogen atoms on a specified atom with 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 of.
  • 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, and -C 0alkyl -A means that the structure is actually -A.
  • 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.
  • a substituent can be bonded to any atom on a ring when the bond of a substituent can cross-link two or more atoms on the ring, e.g., structural unit It means that the substituent R can be substituted at any position on cyclohexyl or cyclohexadiene.
  • 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 1-3 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.
  • halogen or halogen by itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.
  • 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, 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
  • the cultured single crystal is collected with a Bruker D8 venture diffractometer to collect diffraction intensity data
  • the light source is CuK ⁇ radiation
  • 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).
  • the solvent used in the present invention is commercially available.
  • CDCl 3 stands for deuterated chloroform
  • CD 3 OD stands for deuterated methanol
  • DMSO-d 6 stands for deuterated dimethyl sulfoxide.
  • Fig. 1 is the result of immunoblotting experiment on brain tissue of the compound of the present invention.
  • reaction liquid was filtered, and the filtrate was extracted with ethyl acetate (20 ml ⁇ 3), the organic phase was washed with saturated brine (50 ml ⁇ 3), dried over anhydrous sodium sulfate, filtered, and the concentrated crude product was passed through a silica gel column layer.
  • reaction solution was concentrated under reduced pressure, then, water (50 ml) was added, and the pH of the solution was adjusted to 8 with saturated sodium bicarbonate solution, then dichloromethane (50 ml) was added to dilute the reaction solution, and dichloromethane was added to Extracted with methane (50 mL ⁇ 3), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • reaction solution was concentrated under reduced pressure, then, water (50 ml) was added, and the pH of the solution was adjusted to 8 with saturated sodium bicarbonate solution, then dichloromethane (30 ml) was added to dilute the reaction solution, and dichloromethane was added to Extracted with methane (30 mL ⁇ 3), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • reaction solution was filtered, and water (50 milliliters) was added to the filtrate for dilution, and extracted with ethyl acetate (50 milliliters) mL ⁇ 2), the organic phase was washed with saturated brine (50 mL), and dried over anhydrous sodium sulfate.
  • reaction solution was filtered with diatomaceous earth, and the crude product concentrated under reduced pressure of the filtrate was subjected to high performance liquid chromatography (chromatographic column: Waters Xbridge C18 15 ⁇ 50 millimeters ⁇ 10 microns; mobile phase: mobile phase A: ammonium bicarbonate aqueous solution; Mobile phase B: acetonitrile; B%: 40%-70%, 10 minutes) isolated to obtain compound 36.
  • chromatographic column Waters Xbridge C18 15 ⁇ 50 millimeters ⁇ 10 microns
  • mobile phase mobile phase A: ammonium bicarbonate aqueous solution
  • Mobile phase B acetonitrile
  • B% 40%-70%, 10 minutes
  • reaction solution was quenched with saturated ammonium chloride solution (50 mL) at 25°C, diluted with ethyl acetate (50 mL), and extracted with ethyl acetate (40 mL ⁇ 3).
  • the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product.
  • the crude product was separated by high-performance liquid chromatography (chromatographic column: Phenomenex Luna C18 150 ⁇ 40 mm ⁇ 15 microns; mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 20%-50%, 10 minutes) separation Compound 49-5 was obtained.
  • the reaction solution was filtered and concentrated to obtain a crude product.
  • reaction solution was quenched by adding water (20 ml), diluted with ethyl acetate (50 ml), extracted with ethyl acetate (50 ml ⁇ 3), and the combined organic phases were dried over anhydrous sodium sulfate and filtered. , concentrated to obtain the crude product.
  • the reaction solution was filtered and concentrated to obtain a crude product.
  • reaction solution was filtered and concentrated to obtain a crude product.
  • the crude product was subjected to high performance liquid chromatography (chromatographic column: Unisil 3-100C18Ultra 150 ⁇ 50 mm ⁇ 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 44%-74%, 10 minutes ) to isolate compound 51.
  • high-performance liquid chromatography chromatographic column: Phenomenex C18 75 ⁇ 30 mm ⁇ 3 Micron; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 38%-68%, 7 minutes
  • reaction solution was filtered, diluted with water (20 ml), extracted with ethyl acetate (20 ml ⁇ 2), the combined organic phase was washed with saturated saline solution (20 ml), and the organic phase was dried over anhydrous sodium sulfate. , filtered, and the concentrated residue was subjected to high performance liquid chromatography (chromatographic column: YMC Triart C18 150 mm ⁇ 25 mm ⁇ 5 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 59%-89 %, 8.5 min) to isolate compound 56.
  • chromatographic column YMC Triart C18 150 mm ⁇ 25 mm ⁇ 5 microns
  • mobile phase mobile phase A: formic acid aqueous solution
  • mobile phase B acetonitrile
  • B% 59%-89 %, 8.5 min
  • the crude product was separated by high performance liquid chromatography (column: Phenomenex luna C18 150 mm ⁇ 25 mm ⁇ 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 10%-40%) to obtain the compound 60.
  • the purpose of the experiment to detect the phosphorylated Fluorescein-ERM (LRRKtide) peptide phosphate group and Energy signal transfer (520nM/485nM fluorescence signal ratio) generated after Tb-pERM (pLRRKtide) Antibody binding. Calculate the LRRK2 kinase inhibitory IC 50 value of the test compound.
  • Reaction solution 10mM hydroxyethylpiperazineethanesulfonic acid (PH7.5); 2mM magnesium chloride; 0.5mM ethylene glycol diethyl ether diamine tetraacetic acid; 0.002% polyoxyethylene fatty alcohol ether; 1mM dithiothreitol and 1%DMSO;
  • LRRK2 human recombinant protein using GST tag to express recombinant full-length human LRRK2 protein in insect Sf9 cells with baculovirus;
  • Substrate 0.4 ⁇ M Fluorescein-ERM (LRRKtide) peptide; 57 ⁇ M ATP.
  • HTRF Homogeneous Time-Resolved Fluorescence
  • the relative enzyme activity inhibition relative to DMSO blank was calculated by the signal ratio, and the IC 50 value was calculated by using the software XLfit5 to fit the curve.
  • Compound 3 6 Compound 23 6 Compound 4 7 Compound 24 5 Compound 5 5 Compound 25 5 Compound 6 5 Compound 26 5 Compound 7 3 Compound 27 4 Compound 8 4 Compound 28 15 Compound 9 4 Compound 29 14 Compound 10 3 Compound 30 5 Compound 11 14 Compound 31 10 Compound 12 12 Compound 32 7 Compound 15 16 Compound 33 13 Compound 17 5 Compound 36 10 Compound 18 4 Compound 37 7 Compound 19 6 Compound 39 16 Compound 20 4 / /

Abstract

Provided is an azaspiro compound, and specifically an application of a compound represented by formula (II-1), etc. and a pharmaceutically acceptable salt thereof in the preparation for a drug for treating related diseases.

Description

氮杂螺环化合物Azaspiro compounds
本申请主张如下优先权：This application claims the following priority:
CN202111308839.8，申请日2021年11月05日；CN202111308839.8, the application date is November 05, 2021;
CN202210837792.2，申请日2022年07月15日；CN202210837792.2, the application date is July 15, 2022;
CN202211351816.X，申请日2022年10月31日。CN202211351816.X, the application date is October 31, 2022.
技术领域technical field
本发明涉及一类氮杂螺环化合物，具体涉及式(II-1)等所示化合物或其药学上可接受的盐在制备治疗相关疾病药物中的应用。The present invention relates to a class of azaspiro compounds, in particular to the application of compounds represented by formula (II-1) or pharmaceutically acceptable salts thereof in the preparation of drugs for treating related diseases.
背景技术Background technique
LRRK2激酶的突变及过表达越来越多地被证明是诱发神经退行性疾病的根本因素，以黑质区中多巴胺能神经元的选择性变性和细胞死亡为主要特征。影响着1％的65岁以上的人群，其中遗传性患者占发病人群的5-10％。该疾病的早期，最明显的症状表现为摇动,活动缓慢和行走困难。后期还会出现认知和行为问题，晚期通常会出现痴呆。Mutation and overexpression of LRRK2 kinase are increasingly proven to be the fundamental factors in inducing neurodegenerative diseases, characterized by selective degeneration and cell death of dopaminergic neurons in the substantia nigra. Affects 1% of the population over the age of 65, of which genetic patients account for 5-10% of the affected population. Early in the disease, the most obvious symptoms are shaking, slowness of movement and difficulty walking. Cognitive and behavioral problems also develop later, and dementia usually occurs later.
越来越多的证据显示富含亮氨酸重复序列的激酶2(LRRK2)突变与神经退行性疾病有不可分割的联系，LRRK2是一种在催化磷酸化和GTP-GDP水解中涉及的2527氨基酸蛋白。人类LRRK2mRNA的NCBI参与序列是NM_198578.2。证据显示，LRRK2在丝氨酸-129处磷酸化a-突触核蛋白，并且这一磷酸化形式构成路易体的重要部分。另外，已经显示LRRK2的功能结构域中的单核苷酸多肽性引起常见性且散发性神经退行性疾病。目前为止，研究人员已经在患有迟发型神经退行性疾病的家族中识别了超过20个LRRK2突变。例如G2019S突变与常染色体显性共分离，并且其在欧洲导致约6％的家庭性病例和3％散发性病例。G2019S突变发生在高度保守的激酶结构域，因此G2019S突变可能对激酶活性有影响。此外，在另一残基R1441上的氨基酸取代也与神经退行性疾病有关，并且显示提高了LRRK2激酶的活性。在转基因小鼠模型中的突变体LRRK2蛋白R1441G的过度表达与多巴胺释放减少有关，显示LRRK2抑制剂也能积极地调节多巴胺的释放并且在治疗以及多巴胺释放减少有关，显示LRRK2抑制剂也能积极地调节多巴胺的释放并且在治疗以降低的多巴胺水平为特征的疾病中具有潜在效用。相关的数据进一步显示了LRRK2激酶活性抑制剂也可用于治疗相关的神经变性疾病。Accumulating evidence shows that mutations in leucine-rich repeat kinase 2 (LRRK2), a 2527-amino acid protein involved in catalyzing phosphorylation and hydrolysis of GTP-GDP, are inextricably linked to neurodegenerative diseases protein. The NCBI participating sequence for human LRRK2 mRNA is NM_198578.2. Evidence shows that LRRK2 phosphorylates α-synuclein at serine-129 and that this phosphorylated form constitutes an important part of Lewy bodies. In addition, SNPs in the functional domains of LRRK2 have been shown to cause common and sporadic neurodegenerative diseases. So far, researchers have identified more than 20 LRRK2 mutations in families with late-onset neurodegenerative disease. For example the G2019S mutation co-segregates with autosomal dominant and it is responsible for approximately 6% of familial cases and 3% of sporadic cases in Europe. The G2019S mutation occurs in a highly conserved kinase domain, so the G2019S mutation may have an effect on kinase activity. Furthermore, an amino acid substitution at another residue, R1441, has also been associated with neurodegenerative diseases and was shown to increase LRRK2 kinase activity. Overexpression of the mutant LRRK2 protein R1441G in a transgenic mouse model was associated with reduced dopamine release, showing that LRRK2 inhibitors also positively modulate dopamine release and are associated with treatment as well as reduced dopamine release, showing that LRRK2 inhibitors can also positively regulate dopamine release Modulates the release of dopamine and has potential utility in the treatment of diseases characterized by decreased dopamine levels. Related data further demonstrate that inhibitors of LRRK2 kinase activity may also be useful in the treatment of related neurodegenerative diseases.
因此，开发有效的LRRK2激酶以及突变的LRRK2激酶的抑制剂成为目前治疗神经退行性疾病的一条重要的途径。本发明旨在发明一种可以高度对LRRK2激酶抑制的化合物，从而进一步发明可以很好的治疗神经退行性疾病的药物。Therefore, developing effective LRRK2 kinase inhibitors and mutant LRRK2 kinase inhibitors has become an important way to treat neurodegenerative diseases. The present invention aims to invent a compound that can highly inhibit LRRK2 kinase, so as to further invent a drug that can well treat neurodegenerative diseases.
J.Med.Chem.2012,55,9416-9433公开了化合物GNE-7915，属于LRRK2激酶抑制剂，其结构式如下所示：J.Med.Chem.2012,55,9416-9433 discloses compound GNE-7915, which belongs to LRRK2 kinase inhibitor, and its structural formula is as follows:
Figure PCTCN2022129958-appb-000001
Figure PCTCN2022129958-appb-000001
发明内容Contents of the invention
本发明提供式(III-1)、(II-2)、(II-3)、(II-4)和(III-2)所示化合物或其药学上可接受的盐：The present invention provides compounds represented by formulas (III-1), (II-2), (II-3), (II-4) and (III-2) or pharmaceutically acceptable salts thereof:
Figure PCTCN2022129958-appb-000002
Figure PCTCN2022129958-appb-000002
其中，in,
A选自
Figure PCTCN2022129958-appb-000003
Figure PCTCN2022129958-appb-000004
A selected from
Figure PCTCN2022129958-appb-000003
Figure PCTCN2022129958-appb-000004
结构单元
Figure PCTCN2022129958-appb-000005
选自
Figure PCTCN2022129958-appb-000006
Structural units
Figure PCTCN2022129958-appb-000005
selected from
Figure PCTCN2022129958-appb-000006
X 1为CH，X 2为C-A；或者，X 1为C-A，X 2为CH； X 1 is CH, X 2 is CA; or, X 1 is CA, X 2 is CH;
Y为-C(R 9R 10)-或-O-； Y is -C(R 9 R 10 )- or -O-;
Z 1为CH，Z 2为N；或者，Z 1为N，Z 2为CH； Z 1 is CH, Z 2 is N; or, Z 1 is N, Z 2 is CH;
R 1和R 2分别独立地选自H、F、Cl、Br、I、CH 3和CF 3； R 1 and R 2 are independently selected from H, F, Cl, Br, I, CH 3 and CF 3 ;
R 3和R 4分别独立地选自H和C 1-3烷基，所述C 1-3烷基任选被1、2或3个卤素取代； R 3 and R 4 are independently selected from H and C 1-3 alkyl, and the C 1-3 alkyl is optionally substituted by 1, 2 or 3 halogens;
R 5和R 6分别独立地选自H、F、CH 3和CF 3； R 5 and R 6 are independently selected from H, F, CH 3 and CF 3 ;
R 7选自CH 3和CHF 2； R 7 is selected from CH 3 and CHF 2 ;
R 9和R 10分别独立地选自H、F、Cl、Br、I、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基分别独立地任选被1、2或3个卤素取代； R 9 and R 10 are independently selected from H, F, Cl, Br, I, C 1-3 alkyl and C 1-3 alkoxy, the C 1-3 alkyl and C 1-3 alkoxy Each group is independently optionally substituted by 1, 2 or 3 halogens;
m、n、q、t分别独立地选自0、1和2。m, n, q, t are independently selected from 0, 1 and 2, respectively.
在本发明的一些方案中，上述化合物选自式(II-1)、(II-2)、(II-3)和(II-4)所示化合物：In some schemes of the present invention, the above-mentioned compounds are selected from compounds shown in formulas (II-1), (II-2), (II-3) and (II-4):
Figure PCTCN2022129958-appb-000007
Figure PCTCN2022129958-appb-000007
其中，in,
A选自
Figure PCTCN2022129958-appb-000008
A selected from
Figure PCTCN2022129958-appb-000008
X 1为CH，X 2为C-A；或者，X 1为C-A，X 2为CH； X 1 is CH, X 2 is CA; or, X 1 is CA, X 2 is CH;
Y为-C(R 9R 10)-或-O-； Y is -C(R 9 R 10 )- or -O-;
Z 1为CH，Z 2为N；或者，Z 1为N，Z 2为CH； Z 1 is CH, Z 2 is N; or, Z 1 is N, Z 2 is CH;
R 1和R 2分别独立地选自H、F、Cl、Br、I、CH 3和CF 3； R 1 and R 2 are independently selected from H, F, Cl, Br, I, CH 3 and CF 3 ;
R 3和R 4分别独立地选自H和C 1-3烷基，所述C 1-3烷基任选被1、2或3个卤素取代； R 3 and R 4 are independently selected from H and C 1-3 alkyl, and the C 1-3 alkyl is optionally substituted by 1, 2 or 3 halogens;
R 5和R 6分别独立地选自H和F； R 5 and R 6 are independently selected from H and F;
R 7选自CH 3和CHF 2； R 7 is selected from CH 3 and CHF 2 ;
R 9和R 10分别独立地选自H、F、Cl、Br、I、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基分别独立地任选被1、2或3个卤素取代； R 9 and R 10 are independently selected from H, F, Cl, Br, I, C 1-3 alkyl and C 1-3 alkoxy, the C 1-3 alkyl and C 1-3 alkoxy Each group is independently optionally substituted by 1, 2 or 3 halogens;
m、n、q、t分别独立地选自0、1和2。m, n, q, t are independently selected from 0, 1 and 2, respectively.
在本发明的一些方案中，上述化合物选自In some aspects of the present invention, the above compounds are selected from
Figure PCTCN2022129958-appb-000009
Figure PCTCN2022129958-appb-000009
其中，A、R 1、R 2、R 3、R 6和R 7如本发明所定义。 Wherein, A, R 1 , R 2 , R 3 , R 6 and R 7 are as defined in the present invention.
在本发明的一些方案中，上述化合物如式(I-1)、(I-2)、(I-3)、(I-4)、(I-5)和(I-6)所示：In some schemes of the present invention, the above-mentioned compounds are shown in formulas (I-1), (I-2), (I-3), (I-4), (I-5) and (I-6):
Figure PCTCN2022129958-appb-000010
Figure PCTCN2022129958-appb-000010
其中，Y、R 1、R 2、R 3、R 4、m、n、q和t如本发明所定义。 Wherein, Y, R 1 , R 2 , R 3 , R 4 , m, n, q and t are as defined in the present invention.
在本发明的一些方案中，上述式(I-1)、(I-2)、(I-3)、(I-4)、(I-5)和(I-6)所示化合物或其药学上可接受的盐：In some schemes of the present invention, the compounds shown in the above formulas (I-1), (I-2), (I-3), (I-4), (I-5) and (I-6) or Pharmaceutically acceptable salts:
Figure PCTCN2022129958-appb-000011
Figure PCTCN2022129958-appb-000011
其中，in,
Y为-C(R 9R 10)-或-O-； Y is -C(R 9 R 10 )- or -O-;
R 1和R 2分别独立地选自H、F、Cl、Br、I、CH 3和CF 3； R 1 and R 2 are independently selected from H, F, Cl, Br, I, CH 3 and CF 3 ;
R 3和R 4分别独立地选自H和C 1-3烷基，所述C 1-3烷基任选被1、2或3个卤素取代； R 3 and R 4 are independently selected from H and C 1-3 alkyl, and the C 1-3 alkyl is optionally substituted by 1, 2 or 3 halogens;
R 9和R 10分别独立地选自H、F、Cl、Br、I、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基分别独立地任选被1、2或3个卤素取代； R 9 and R 10 are independently selected from H, F, Cl, Br, I, C 1-3 alkyl and C 1-3 alkoxy, the C 1-3 alkyl and C 1-3 alkoxy Each group is independently optionally substituted by 1, 2 or 3 halogens;
m、n、q、t分别独立地选自0、1和2。m, n, q, t are independently selected from 0, 1 and 2, respectively.
在本发明的一些方案中，上述R 1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 1 is selected from H and F, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 2选自Cl和F，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 Cl and F, and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 3和R 4分别独立地选自CH 3，其他变量如本发明所定义。 In some solutions of the present invention, the above R 3 and R 4 are independently selected from CH 3 , and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 9和R 10分别独立地选自H、F、CH 3、-OCHF 2和-OCH 3，其他变量如本发明所定义。 In some solutions of the present invention, the above R 9 and R 10 are independently selected from H, F, CH 3 , -OCHF 2 and -OCH 3 , and other variables are as defined in the present invention.
在本发明的一些方案中，上述R 9和R 10分别独立地选自H、F和-OCH 3，其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned R 9 and R 10 are independently selected from H, F and -OCH 3 , and other variables are as defined in the present invention.
在本发明的一些方案中，上述Y为-CHF-、-CF 2-、-CH 2-、-CH(OCH 3)-、-CH(OCHF 2)-和-O-，其他 变量如本发明所定义。 In some solutions of the present invention, the above-mentioned Y is -CHF-, -CF 2 -, -CH 2 -, -CH(OCH 3 )-, -CH(OCHF 2 )- and -O-, other variables are as in the present invention defined.
在本发明的一些方案中，上述结构单元
Figure PCTCN2022129958-appb-000012
选自
Figure PCTCN2022129958-appb-000013
Figure PCTCN2022129958-appb-000014
Figure PCTCN2022129958-appb-000015
其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned structural units
Figure PCTCN2022129958-appb-000012
selected from
Figure PCTCN2022129958-appb-000013
Figure PCTCN2022129958-appb-000014
Figure PCTCN2022129958-appb-000015
Other variables are as defined herein.
在本发明的一些方案中，上述结构单元
Figure PCTCN2022129958-appb-000016
选自
Figure PCTCN2022129958-appb-000017
其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned structural units
Figure PCTCN2022129958-appb-000016
selected from
Figure PCTCN2022129958-appb-000017
Other variables are as defined herein.
在本发明的一些方案中，上述结构单元
Figure PCTCN2022129958-appb-000018
选自
Figure PCTCN2022129958-appb-000019
其他变量如本发明所定义。 In some solutions of the present invention, the above-mentioned structural units
Figure PCTCN2022129958-appb-000018
selected from
Figure PCTCN2022129958-appb-000019
Other variables are as defined herein.
在本发明的一些方案中，上述式(II-2)中的结构单元
Figure PCTCN2022129958-appb-000020
选自
Figure PCTCN2022129958-appb-000021
Figure PCTCN2022129958-appb-000022
其他变量如本发明所定义。 In some schemes of the present invention, the structural unit in the above formula (II-2)
Figure PCTCN2022129958-appb-000020
selected from
Figure PCTCN2022129958-appb-000021
Figure PCTCN2022129958-appb-000022
Other variables are as defined herein.
在本发明的一些方案中，上述式(III-2)中的结构单元
Figure PCTCN2022129958-appb-000023
选自
Figure PCTCN2022129958-appb-000024
其他变量如本发明所定义。 In some schemes of the present invention, the structural unit in the above formula (III-2)
Figure PCTCN2022129958-appb-000023
selected from
Figure PCTCN2022129958-appb-000024
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.
本发明还提供化合物或其药学上可接受的盐，其中，化合物选自The present invention also provides a compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from
Figure PCTCN2022129958-appb-000025
Figure PCTCN2022129958-appb-000025
Figure PCTCN2022129958-appb-000026
Figure PCTCN2022129958-appb-000026
Figure PCTCN2022129958-appb-000027
Figure PCTCN2022129958-appb-000027
Figure PCTCN2022129958-appb-000028
Figure PCTCN2022129958-appb-000028
Figure PCTCN2022129958-appb-000029
Figure PCTCN2022129958-appb-000029
本发明还提供上述化合物或其药学上可接受的盐在制备治疗帕金森病药物中的应用。The present invention also provides the application of the above compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating Parkinson's disease.
技术效果technical effect
本发明化合物对LRRK2具有显著的激酶抑制和细胞活性以及透膜性和溶解性，同时具有优良的药代动力学和药效学性质，如具有较好的入脑性质。The compound of the present invention has remarkable kinase inhibition, cell activity, membrane permeability and solubility for LRRK2, and has excellent pharmacokinetic and pharmacodynamic properties, such as better brain entry properties.
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 name or its active ingredient.
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 and the like , 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.
除非另有规定，术语“有效量”或“治疗有效量”是指无毒的但能达到预期效果的用量。有效量的确定因人而异，取决于受体的年龄和一般情况，也取决于具体的活性物质，个案中合适的有效量可以由本领域技术人员根据常规试验确定。Unless otherwise specified, the terms "effective amount" or "therapeutically effective amount" refer to a non-toxic amount that achieves the desired effect. The determination of the effective amount varies from person to person, depending on the age and general condition of the recipient, and also depends on the specific active substance. The appropriate effective amount in each case can be determined by those skilled in the art according to routine experiments.
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，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。The term "substituted" refers to the replacement of any one or more hydrogen atoms on a specified atom with 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 of. When a substituent is oxygen (ie =0), it means that two hydrogen atoms are replaced.
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，-C 0烷基-A表示该结构实际上是-A。 When the number of a linking group is 0, such as -(CRR) 0 -, it means that the linking group is a single bond, and -C 0alkyl -A means that the structure is actually -A.
当一个取代基数量为0时，表示该取代基是不存在的，比如-A-(R) 0表示该结构实际上是-A。 When the number of a substituent is 0, it means that the substituent does not exist, such as -A-(R) 0 means that the structure is actually -A.
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。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.
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。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.
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 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.
当一个取代基的键可以交叉连接到一个环上的两一个以上原子时，这种取代基可以与这个环上的任意原子相键合，例如，结构单元
Figure PCTCN2022129958-appb-000030
表示其取代基R可在环己基或者环己二烯上的任意一个位置发生取代。 A substituent can be bonded to any atom on a ring when the bond of a substituent can cross-link two or more atoms on the ring, e.g., structural unit
Figure PCTCN2022129958-appb-000030
It means that the substituent R can be substituted at any position on cyclohexyl or cyclohexadiene.
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 PCTCN2022129958-appb-000031
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 PCTCN2022129958-appb-000032
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 PCTCN2022129958-appb-000033
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 PCTCN2022129958-appb-000031
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 PCTCN2022129958-appb-000032
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 PCTCN2022129958-appb-000033
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 PCTCN2022129958-appb-000034
直形虚线键
Figure PCTCN2022129958-appb-000035
或波浪线
Figure PCTCN2022129958-appb-000036
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 PCTCN2022129958-appb-000037
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 PCTCN2022129958-appb-000038
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 PCTCN2022129958-appb-000039
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 PCTCN2022129958-appb-000040
这4种连接方式，即使-N-上画出了H原子，但是
Figure PCTCN2022129958-appb-000041
仍包括
Figure PCTCN2022129958-appb-000042
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 PCTCN2022129958-appb-000034
Straight dotted key
Figure PCTCN2022129958-appb-000035
or tilde
Figure PCTCN2022129958-appb-000036
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 PCTCN2022129958-appb-000037
The straight dotted line bond in indicates that the two ends of the nitrogen atom in the group are connected to other groups;
Figure PCTCN2022129958-appb-000038
The wavy lines in indicate that the 1 and 2 carbon atoms in the phenyl group are connected to other groups;
Figure PCTCN2022129958-appb-000039
Indicates that any connectable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least
Figure PCTCN2022129958-appb-000040
These 4 connection methods, even if the H atom is drawn on -N-, but
Figure PCTCN2022129958-appb-000041
still include
Figure PCTCN2022129958-appb-000042
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 1-3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“卤代素”或“卤素”本身或作为另一取代基的一部分表示氟、氯、溴或碘原子。Unless otherwise specified, the term "halogen" or "halogen" by itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.
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 PCTCN2022129958-appb-000043
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, 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 PCTCN2022129958-appb-000043
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。The solvent used in the present invention is commercially available.
本发明采用下述缩略词：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：CDCl 3代表氘代氯仿，CD 3OD代表氘代甲醇，DMSO-d 6代表氘代二甲亚砜。 The following abbreviations are used in the present invention: The solvents used in the present invention are commercially available. The following abbreviations are used in the present invention: CDCl 3 stands for deuterated chloroform, CD 3 OD stands for deuterated methanol, and DMSO-d 6 stands for deuterated dimethyl sulfoxide.
化合物依据本领域常规命名原则或者使用
Figure PCTCN2022129958-appb-000044
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 PCTCN2022129958-appb-000044
The software is named, and the commercially available compounds adopt the supplier catalog name.
附图说明Description of drawings
图1为本发明化合物脑组织免疫印迹实验结果。Fig. 1 is the result of immunoblotting experiment on brain tissue of the compound of the present invention.
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。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 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. Various changes and modifications to the specific embodiments of the invention will be apparent to those skilled in the art without departing from the spirit and scope of the invention.
中间体A制备Preparation of Intermediate A
Figure PCTCN2022129958-appb-000045
Figure PCTCN2022129958-appb-000045
第一步first step
将化合物A-1(400毫克，1.76毫摩尔,1当量)溶于无水甲醇(10毫升)，接着向反应液中加入氯化氢的甲醇溶液(2毫升，4摩尔/升，3当量)，在25摄氏度氮气保护下反应2小时。反应完成后，减压浓缩纯化得到中间体A。Compound A-1 (400 mg, 1.76 mmol, 1 equivalent) was dissolved in anhydrous methanol (10 milliliters), and then a methanolic solution of hydrogen chloride (2 milliliters, 4 mol/liter, 3 equivalents) was added to the reaction solution. React for 2 hours under nitrogen protection at 25°C. After the reaction was completed, intermediate A was obtained by concentration and purification under reduced pressure.
MS-ESI计算值[M+H] +128,实测值128。 MS-ESI calculated value [M+H] + 128, found value 128.
中间体B制备Preparation of Intermediate B
Figure PCTCN2022129958-appb-000046
Figure PCTCN2022129958-appb-000046
第一步first step
将化合物B-1(20克，93.78毫摩尔，1当量)溶于二氯甲烷(200毫升)，接着在0摄氏度下向反应液中加入三氟化二乙氨基硫(18.14克，112.53毫摩尔，14.87毫升，1.2当量)，然后在40摄氏度下反应12小时。反应完成后，加入200毫升水淬灭反应，然后用二氯甲烷(100毫升×3)萃取，再用饱和食盐水(100毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤后，减压浓缩得到粗品经硅胶柱层析(石油醚：乙酸乙酯＝25：1-10：1)纯化得到粗品化合物B-2。Compound B-1 (20 g, 93.78 mmol, 1 equivalent) was dissolved in dichloromethane (200 ml), and then diethylaminosulfur trifluoride (18.14 g, 112.53 mmol) was added to the reaction solution at 0° C. , 14.87 ml, 1.2 equivalents), then reacted at 40 degrees Celsius for 12 hours. After the reaction was complete, 200 ml of water was added to quench the reaction, then extracted with dichloromethane (100 ml×3), washed with saturated brine (100 ml×2), the organic phase was dried over anhydrous sodium sulfate, and filtered, The crude product was concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 25:1-10:1) to obtain the crude compound B-2.
第二步second step
将化合物B-2(300毫克，1.39毫摩尔，1当量)溶于乙酸乙酯(3毫升)，接着在室温下向反应液中加入氯化氢的乙酸乙酯(4摩尔/升，3.31毫升，8.61当量)，在25摄氏度下反应2小时。反应完成后，加入碳酸氢钠饱和水溶液(10毫升)，二氯甲烷萃取(10毫升×3)，有机相经饱和食盐水(10毫升×3)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩。粗品未经纯化得到中间体B。Compound B-2 (300 mg, 1.39 mmol, 1 eq) was dissolved in ethyl acetate (3 ml), then ethyl acetate in hydrogen chloride (4 mol/L, 3.31 ml, 8.61 Equivalent), reacted at 25 degrees Celsius for 2 hours. After the reaction was complete, add saturated aqueous sodium bicarbonate (10 ml), extract with dichloromethane (10 ml × 3), wash the organic phase with saturated brine (10 ml × 3), dry the organic phase with anhydrous sodium sulfate, and filter ,concentrate. Intermediate B was obtained from the crude product without purification.
MS-ESI计算值[M+H] +116,实测值116。 MS-ESI calculated value [M+H] + 116, found value 116.
中间体C制备Preparation of Intermediate C
Figure PCTCN2022129958-appb-000047
Figure PCTCN2022129958-appb-000047
第一步first step
将化合物C-1(350毫克，1.50毫摩尔，1当量)溶于无水甲醇(10毫升)，接着向反应液中加入氯化氢的甲醇溶液(2毫升，4摩尔/升，3当量)，在25摄氏度氮气保护下反应2小时。反应完成后，减压浓缩 纯化得到中间体C。Compound C-1 (350 mg, 1.50 mmol, 1 equivalent) was dissolved in anhydrous methanol (10 milliliters), and then a methanolic solution of hydrogen chloride (2 milliliters, 4 mol/liter, 3 equivalents) was added to the reaction solution. React for 2 hours under nitrogen protection at 25°C. After completion of the reaction, intermediate C was obtained by concentration and purification under reduced pressure.
MS-ESI计算值[M+H] +134,实测值134。 MS-ESI calculated value [M+H] + 134, found value 134.
实施例1Example 1
Figure PCTCN2022129958-appb-000048
Figure PCTCN2022129958-appb-000048
第一步first step
将化合物1-1(5.00克，24.75毫摩尔，1当量)溶于四氢呋喃(50毫升)，接着在25摄氏度下向反应液中加入二碳酸二叔丁酯(8.10克，37.12毫摩尔，8.53毫升，1.5当量)，在70摄氏度下反应10小时。反应完成后，过滤，减压浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1-20：1)纯化得到粗品化合物1-2。Compound 1-1 (5.00 g, 24.75 mmol, 1 equivalent) was dissolved in tetrahydrofuran (50 ml), and then di-tert-butyl dicarbonate (8.10 g, 37.12 mmol, 8.53 ml) was added to the reaction solution at 25 degrees Celsius. , 1.5 equivalents), reacted at 70 degrees Celsius for 10 hours. After the reaction was completed, it was filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1-20:1) to obtain crude compound 1-2.
MS-ESI计算值[M+H] +302，实测值302。 MS-ESI calculated [M+H] + 302, found 302.
第二步second step
将化合物1-2(800毫克，2.65毫摩尔，1当量)溶于二氧六环(40毫升)，接着向反应液中加入碳酸铯(2.59毫克，7.94毫摩尔，3当量)和中间体D(474.94毫克，3.97毫摩尔，1.5当量)，4,5-双二苯基膦-99-二甲基氧杂氧杂蒽杂蒽(306.39毫克，529.51微摩尔，0.2当量)和三(二亚苄基丙酮)二钯(484.89毫克，529.51毫摩尔，0.2当量)，然后置换氮气三次。并在氮气氛围下在100摄氏度下反应12小时。反应完成后，过滤，减压浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1-25：1)纯化得到化合物1-3。Compound 1-2 (800 mg, 2.65 mmol, 1 eq) was dissolved in dioxane (40 mL), then cesium carbonate (2.59 mg, 7.94 mmol, 3 eq) and intermediate D were added to the reaction solution (474.94 mg, 3.97 mmol, 1.5 equiv), 4,5-bisdiphenylphosphine-99-dimethyloxaxanthene (306.39 mg, 529.51 micromol, 0.2 equiv) and tris(diphenyl benzylacetone) dipalladium (484.89 mg, 529.51 mmol, 0.2 equiv), and then replace nitrogen three times. And react at 100 degrees Celsius for 12 hours under nitrogen atmosphere. After the reaction was completed, it was filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1-25:1) to obtain compound 1-3.
MS-ESI计算值[M+H] +305，实测值305。 MS-ESI calculated [M+H] +305 , found 305.
第三步third step
将化合物1-3(456毫克，1.50毫摩尔，1当量)溶于二氯甲烷(5毫升)，接着向反应液中加入三氟乙酸(7.70克，67.53毫摩尔，5毫升，45.08当量)，在25摄氏度下反应0.5小时。反应完成后，减压浓缩，将反应液加入饱和的碳酸氢钠溶液(20毫升)，用二氯甲烷萃取(20毫升×2)，分液，有机相用无水硫酸钠干燥，过滤，减压浓缩。粗品经硅胶薄层色谱板纯化(石油醚：乙酸乙酯＝5:2)得到化合物1-4。Compound 1-3 (456 mg, 1.50 mmol, 1 eq) was dissolved in dichloromethane (5 mL), then trifluoroacetic acid (7.70 g, 67.53 mmol, 5 mL, 45.08 eq) was added to the reaction solution, React at 25 degrees Celsius for 0.5 hours. After completion of the reaction, concentrate under reduced pressure, add the reaction solution to saturated sodium bicarbonate solution (20 ml), extract with dichloromethane (20 ml × 2), separate the layers, dry the organic phase with anhydrous sodium sulfate, filter, and reduce Concentrate under pressure. The crude product was purified by silica gel thin-layer chromatography (petroleum ether: ethyl acetate = 5:2) to obtain compound 1-4.
MS-ESI计算值[M+H] +205，实测值205。 MS-ESI calculated [M+H] +205 , found 205.
第四步the fourth step
将化合物1-4(110毫克，538.51毫摩尔，1当量)溶于四氢呋喃(15毫升)，接着向反应液中加入叔丁醇钠(103.51毫克，1.08毫摩尔，2当量)，中间体E(170.23毫克，538.51微摩尔，1当量)，2-二环己基磷-2,4,6-三异丙基联苯(51.34毫克，107.70微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(49.31毫克， 53.85微摩尔，0.1当量)，置换三次氮气，并在氮气氛围下在70摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝50/1～二氯甲烷：甲醇＝20：1)纯化得到的粗品经高效液相色谱(色谱柱：Waters Xbridge 150*25*10微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％:36％-66％，10分钟)分离得到化合物1。Compound 1-4 (110 mg, 538.51 mmol, 1 eq) was dissolved in tetrahydrofuran (15 ml), then sodium tert-butoxide (103.51 mg, 1.08 mmol, 2 eq) was added to the reaction solution, Intermediate E ( 170.23 mg, 538.51 micromoles, 1 equiv), 2-dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (51.34 mg, 107.70 micromoles, 0.2 equiv), tris(dibenzylideneacetone) Dipalladium (49.31 mg, 53.85 micromole, 0.1 eq.) was replaced with nitrogen three times, and reacted at 70° C. for 12 hours under nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 50/1 ~ dichloromethane: methanol = 20: 1) and the crude product was purified by high performance liquid chromatography (column: Waters Xbridge 150*25*10 microns; Mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 36%-66%, 10 minutes) to separate and obtain compound 1.
MS-ESI计算值[M+H] +484,实测值484. MS-ESI calculated value [M+H] + 484, measured value 484.
1H NMR(400MHz,CD 3OD)δ＝0.64-0.69(m,4H),1.80-1.90(m,6H),3.64-3.71(m,3H),3.95-4.01(m,4H),6.24-6.32(m,1H),6.79-6.87(m,1H),7.27-7.33(m,1H),7.38-7.43(m,1H),7.53-7.59(m,1H),7.60-7.68(m,1H),8.07-8.13(m,1H),8.30-8.38(m,1H). 1 H NMR (400MHz, CD 3 OD) δ=0.64-0.69(m,4H),1.80-1.90(m,6H),3.64-3.71(m,3H),3.95-4.01(m,4H),6.24- 6.32(m,1H),6.79-6.87(m,1H),7.27-7.33(m,1H),7.38-7.43(m,1H),7.53-7.59(m,1H),7.60-7.68(m,1H ),8.07-8.13(m,1H),8.30-8.38(m,1H).
实施例2Example 2
Figure PCTCN2022129958-appb-000049
Figure PCTCN2022129958-appb-000049
第一步first step
将化合物1-2(1克，3.31毫摩尔，1当量)溶于二氧六环(30毫升)，接着向反应液中加入碳酸铯(3.23克，9.93毫摩尔，3当量)和中间体B(752.62毫克，4.96毫摩尔，1.5当量)，45-双二苯基膦-99-二甲基氧杂氧杂蒽杂蒽(382.98毫克，661.89微摩尔，0.2当量)和三(二亚苄基丙酮)二钯(303.05毫克，330.95毫摩尔，0.1当量)，然后置换氮气三次。并在氮气氛围下在100摄氏度下反应12小时。反应完成后，过滤，减压浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1-25：1)纯化得到粗品化合物2-1。Compound 1-2 (1 g, 3.31 mmol, 1 eq) was dissolved in dioxane (30 ml), then cesium carbonate (3.23 g, 9.93 mmol, 3 eq) and intermediate B were added to the reaction solution (752.62mg, 4.96mmol, 1.5eq), 45-bisdiphenylphosphine-99-dimethyloxaxanthene (382.98mg, 661.89mmol, 0.2eq) and tris(dibenzylidene acetone) dipalladium (303.05 mg, 330.95 mmol, 0.1 equiv), and then replace nitrogen three times. And react at 100 degrees Celsius for 12 hours under nitrogen atmosphere. After the reaction was completed, it was filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1-25:1) to obtain crude compound 2-1.
第二步second step
将化合物2-1(669毫克，1.99毫摩尔，1当量)溶于二氯甲烷(6毫升)，接着向反应液中加入三氟乙酸(9.24克，81.04毫摩尔，6毫升，40.75当量)，在25摄氏度下反应0.5小时。反应完成后，过滤，减压浓缩。粗品经硅胶薄层色谱板纯化(石油醚：乙酸乙酯＝50：1-5：1)得到化合物2-2。Compound 2-1 (669 mg, 1.99 mmol, 1 eq) was dissolved in dichloromethane (6 mL), then trifluoroacetic acid (9.24 g, 81.04 mmol, 6 mL, 40.75 eq) was added to the reaction solution, React at 25 degrees Celsius for 0.5 hours. After the reaction was completed, it was filtered and concentrated under reduced pressure. The crude product was purified by silica gel thin-layer chromatography (petroleum ether: ethyl acetate = 50:1-5:1) to obtain compound 2-2.
第三步third step
将化合物2-2(150毫克，634.83毫摩尔，1当量)溶于四氢呋喃(30毫升)，接着向反应液中加入叔丁醇钠(122.02毫克，1.27毫摩尔，2当量)，中间体E(220.75毫克，698.31微摩尔，1.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(60.53毫克，126.97微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(58.13毫克，63.48微摩尔，0.1当量)，置换三次氮气，并在氮气氛围下在70摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝50/1～二氯甲烷：甲醇＝20：1)纯化得到的粗品经高效液相色谱(色谱柱：Waters Xbridge 150*50毫米*3微米；流动相：流动相A：甲酸溶液；流动相 B：乙腈；B％:35％-65％，10分钟)分离得到化合物2。Compound 2-2 (150 mg, 634.83 mmol, 1 eq) was dissolved in tetrahydrofuran (30 ml), then sodium tert-butoxide (122.02 mg, 1.27 mmol, 2 eq) was added to the reaction solution, Intermediate E ( 220.75 mg, 698.31 micromoles, 1.1 equiv), 2-dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (60.53 mg, 126.97 micromoles, 0.2 equiv), tris(dibenzylideneacetone) Dipalladium (58.13 mg, 63.48 micromole, 0.1 eq.) was replaced with nitrogen three times, and reacted at 70° C. for 12 hours under nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate=50/1～dichloromethane:methanol=20:1) and the crude product obtained by high performance liquid chromatography (chromatographic column: Waters Xbridge 150*50 mm*3 microns ; Mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 35%-65%, 10 minutes) separation to obtain compound 2.
MS-ESI计算值[M+H] +516,实测值516. MS-ESI calculated value [M+H] + 516, measured value 516.
1H NMR(400MHz,CD 3OD)δ＝1.82-1.87(m,6H),2.25-2.48(m,2H),2.51-2.71(m,2H),3.63-3.67(m,3H),3.84-3.88(m,4H),6.18-6.27(m,1H),6.77-6.86(m,1H),7.25-7.34(m,1H),7.38-7.43(m,1H),7.49-7.58(m,1H),7.59-7.68(m,1H),8.07-8.12(m,1H),8.29-8.36(m,1H). 1 H NMR (400MHz, CD 3 OD) δ=1.82-1.87(m,6H),2.25-2.48(m,2H),2.51-2.71(m,2H),3.63-3.67(m,3H),3.84- 3.88(m,4H),6.18-6.27(m,1H),6.77-6.86(m,1H),7.25-7.34(m,1H),7.38-7.43(m,1H),7.49-7.58(m,1H ),7.59-7.68(m,1H),8.07-8.12(m,1H),8.29-8.36(m,1H).
实施例3Example 3
Figure PCTCN2022129958-appb-000050
Figure PCTCN2022129958-appb-000050
第一步first step
将化合物1-2(1.00克，3.31毫摩尔，1当量)和中间体F(591.30毫克，3.80毫摩尔，1.15当量)溶于二氧六环(20毫升)，接着向反应液中加入碳酸铯(3.23克，9.93毫摩尔，3当量)、三(二亚苄基丙酮)二钯(303.05毫克，330.95微摩尔，0.1当量)、4,5-双(二苯基膦)-9，9-二甲基氧杂蒽(191.49毫克，330.95微摩尔，0.1当量)在100摄氏度氮气保护下反应12小时。反应完成后，加水(100毫升)，乙酸乙酯萃取(60毫升×3)，有机相用饱和食盐水(60毫升×3)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1到20：1)纯化得到化合物3-1。Compound 1-2 (1.00 g, 3.31 mmol, 1 eq) and Intermediate F (591.30 mg, 3.80 mmol, 1.15 eq) were dissolved in dioxane (20 mL), then cesium carbonate was added to the reaction solution (3.23 g, 9.93 mmol, 3 equiv), tris(dibenzylideneacetone) dipalladium (303.05 mg, 330.95 micromol, 0.1 equiv), 4,5-bis(diphenylphosphine)-9,9- Dimethylxanthene (191.49 mg, 330.95 micromole, 0.1 equiv) was reacted at 100°C under nitrogen protection for 12 hours. After the reaction was complete, add water (100 ml), extract with ethyl acetate (60 ml × 3), wash the organic phase with saturated brine (60 ml × 3), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1 to 20:1) to obtain compound 3-1.
MS-ESI计算值[M+H] +341,实测值341。 MS-ESI calculated [M+H] + 341, found 341.
1H NMR(400MHz,CDCl 3)δ＝8.12-8.03(m,1H),7.65-7.59(m,2H),7.46-7.40(m,1H),7.01–6.95(m,2H),6.26-6.18(m,2H),5.40-5.25(m,3H),4.19-4.09(m,5H),4.02-3.93(m,5H),3.90-3.83(m,1H),3.71-3.63(m,5H),1.50-1.40(m,9H),0.97-0.84(m,1H)。 1 H NMR (400MHz, CDCl 3 )δ=8.12-8.03(m,1H),7.65-7.59(m,2H),7.46-7.40(m,1H),7.01–6.95(m,2H),6.26-6.18 (m,2H),5.40-5.25(m,3H),4.19-4.09(m,5H),4.02-3.93(m,5H),3.90-3.83(m,1H),3.71-3.63(m,5H) ,1.50-1.40(m,9H),0.97-0.84(m,1H).
第二步second step
将化合物3-1(400.00毫克，1.18毫摩尔，1当量)溶于二氯甲烷(4毫升)，向反应液中加入三氟乙酸(369.07毫克，870.15微摩尔，2.5当量)，在25摄氏度下反应0.5小时。反应完成后，加入水(10毫升)，乙酸乙酯萃取(10毫升×3)，有机相经饱和食盐水(10毫升×3)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩。减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝80：1到10：1)纯化得到化合物3-2。Compound 3-1 (400.00 mg, 1.18 mmol, 1 equivalent) was dissolved in dichloromethane (4 ml), and trifluoroacetic acid (369.07 mg, 870.15 micromol, 2.5 equivalents) was added to the reaction solution, and at 25 degrees Celsius React for 0.5 hours. After the reaction was complete, water (10 mL) was added, extracted with ethyl acetate (10 mL×3), the organic phase was washed with saturated brine (10 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue concentrated under reduced pressure was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 80:1 to 10:1) to obtain compound 3-2.
MS-ESI计算值[M+H] +241,实测值241。 MS-ESI calculated value [M+H] + 241, found value 241.
第三步third step
将化合物3-2(160毫克，665.98微摩尔，1.0当量)，中间体E(231.58毫克，732.57微摩尔，1.1 当量)，叔丁醇钠(192.01毫克，2.00毫摩尔，3.0当量)，2-二环己基磷-2,4,6-三异丙基联苯(63.50毫克，133.20微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(60.98毫克，66.60微摩尔，0.1当量)加入四氢呋喃(20毫升)中，氮气置换保护，反应液加热到70摄氏下反应12小时。反应完成后，过滤，滤液减压浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝15:1)和高效液相色谱(色谱柱：Phenomenex luna C18 150*25毫米*10微米；流动相：流动相A：体积分数0.225％甲酸水溶液；流动相B：乙腈；B％:41％-71％,10分钟)纯化得到化合物3。Compound 3-2 (160 mg, 665.98 micromoles, 1.0 equivalents), Intermediate E (231.58 mgs, 732.57 micromoles, 1.1 equivalents), sodium tert-butoxide (192.01 mg, 2.00 mmoles, 3.0 equivalents), 2- Dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (63.50 mg, 133.20 micromoles, 0.2 equiv), tris(dibenzylideneacetone)dipalladium (60.98 mg, 66.60 micromoles, 0.1 equiv) Tetrahydrofuran (20 ml) was added, nitrogen replacement protection was carried out, and the reaction solution was heated to 70° C. for 12 hours. After the reaction was completed, filter, and the residue of the filtrate concentrated under reduced pressure was subjected to silica gel thin-layer chromatography (dichloromethane: methanol=15:1) and high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150*25 millimeters*10 microns; mobile Phase: mobile phase A: 0.225% formic acid aqueous solution by volume fraction; mobile phase B: acetonitrile; B%: 41%-71%, 10 minutes) purification to obtain compound 3.
MS-ESI计算值[M+H] +520,实测值520。 MS-ESI calculated value [M+H] + 520, found value 520.
1HNMR(400MHz,CD 3OD)δ＝8.38-8.30(m,1H),8.12(s,1H),7.70-7.53(m,2H),7.49-7.42(m,1H),7.35-7.26(m,1H),6.91-6.83(m,1H),6.36-6.29(m,1H),4.14-4.08(m,2H),4.01-3.94(m,2H),3.69(s,3H),1.88(s,3H),1.85(s,3H),1.59-1.51(m,2H)。 1 HNMR (400MHz, CD 3 OD)δ=8.38-8.30(m,1H),8.12(s,1H),7.70-7.53(m,2H),7.49-7.42(m,1H),7.35-7.26(m ,1H),6.91-6.83(m,1H),6.36-6.29(m,1H),4.14-4.08(m,2H),4.01-3.94(m,2H),3.69(s,3H),1.88(s ,3H), 1.85(s,3H), 1.59-1.51(m,2H).
实施例4Example 4
Figure PCTCN2022129958-appb-000051
Figure PCTCN2022129958-appb-000051
第一步first step
将化合物1-2(0.9克，2.98毫摩尔)溶于二氧六环(10毫升)，加入中间体A(894.03毫克，4.47毫摩尔)，碳酸铯(2.91克，8.94毫摩尔)，4,5-双二苯基膦-9,9-二甲基氧杂氧杂蒽杂蒽(344.68毫克，595.70微摩尔)和三(二亚苄基丙酮)二钯(545.50毫克，595.70微摩尔)，用氮气置换三次，反应液在氮气保护下升温到100摄氏度，反应12小时。反应完毕后，反应液过滤，浓缩。粗品经柱层析纯化(石油醚：乙酸乙酯＝100:1～10:1)得到化合物4-1。Compound 1-2 (0.9 g, 2.98 mmol) was dissolved in dioxane (10 mL), Intermediate A (894.03 mg, 4.47 mmol), cesium carbonate (2.91 g, 8.94 mmol), 4, 5-bisdiphenylphosphine-9,9-dimethyloxaxanthene (344.68 mg, 595.70 micromoles) and tris(dibenzylideneacetone)dipalladium (545.50 mg, 595.70 micromoles), Replaced with nitrogen three times, the temperature of the reaction solution was raised to 100 degrees Celsius under the protection of nitrogen, and reacted for 12 hours. After the reaction was completed, the reaction solution was filtered and concentrated. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 100:1-10:1) to obtain compound 4-1.
MS-ESI计算值[M+H] +349，实测值349。 MS-ESI calculated [M+H] + 349, found 349.
第二步second step
将化合物4-1(320毫克，918.39微摩尔)溶于二氯甲烷(5毫升)，加入三氟乙酸(7.7克，67.53毫摩尔，5毫升)，反应液在20摄氏度下反应0.5小时。反应完毕后，反应液浓缩，用饱和碳酸氢钠溶液调节pH到8，二氯甲烷(10毫升×2)萃取，有机相用硫酸钠干燥，过滤，浓缩，得到化合物4-2。Compound 4-1 (320 mg, 918.39 micromole) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (7.7 g, 67.53 mmol, 5 mL) was added, and the reaction solution was reacted at 20 degrees Celsius for 0.5 hours. After the reaction was completed, the reaction solution was concentrated, adjusted to pH 8 with saturated sodium bicarbonate solution, extracted with dichloromethane (10 ml×2), and the organic phase was dried over sodium sulfate, filtered, and concentrated to obtain compound 4-2.
MS-ESI计算值[M+H] +249，实测值249。 MS-ESI calculated [M+H] + 249, found 249.
第三步third step
将化合物4-2(110毫克，442.98微摩尔)溶于四氢呋喃(20毫升)，加入化合物中间体E(140.03毫克，442.98微摩尔)，叔丁醇钠(85.14毫克，885.95毫摩尔)，三(二亚苄基丙酮)二钯(40.56毫克， 44.30微摩尔)和2-二环己基磷-2,4,6-三异丙基联苯(42.23毫克，88.60微摩尔)和，用氮气置换三次，反应液在氮气保护下升温到70摄氏度，反应12小时。反应完毕后，反应液过滤，浓缩。粗品经硅胶薄层色谱板纯化(二氯甲烷：甲醇＝10:1)后，经高效液相色谱(色谱柱：Waters Xbridge 150*25毫米*微米；流动相：流动相A：氨水溶液；流动相B：乙腈；B％:65％-65％，10分钟)分离得到化合物4。Compound 4-2 (110 mg, 442.98 micromol) was dissolved in tetrahydrofuran (20 ml), compound intermediate E (140.03 mg, 442.98 micromol), sodium tert-butoxide (85.14 mg, 885.95 mmol), tri( Dibenzylideneacetone) dipalladium (40.56 mg, 44.30 micromoles) and 2-dicyclohexylphospho-2,4,6-triisopropylbiphenyl (42.23 mg, 88.60 micromoles) and, three times replaced with nitrogen , the temperature of the reaction solution was raised to 70° C. under the protection of nitrogen, and the reaction was carried out for 12 hours. After the reaction was completed, the reaction solution was filtered and concentrated. After the crude product is purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1), it is subjected to high performance liquid chromatography (column: Waters Xbridge 150*25 mm*micron; mobile phase: mobile phase A: ammonia solution; mobile phase Phase B: acetonitrile; B%: 65%-65%, 10 minutes) Compound 4 was isolated.
MS-ESI计算值[M+H] +528，实测值528。 MS-ESI calculated [M+H] + 528, found 528.
1H NMR(400MHz,CD 3OD)δ＝1.78-1.84(m,3H),1.84-1.87(m,3H),2.04-2.12(m,2H),2.45-2.55(m,2H),3.20-3.25(m,3H),3.58-3.67(m,3H),3.77-3.81(m,2H),3.81-3.89(m,3H),6.18-6.25(m,1H),6.76-6.83(m,1H),7.24-7.31(m,1H),7.37-7.42(m,1H),7.50-7.57(m,1H),7.58-7.67(m,1H),8.07-8.11(m,1H),8.29-8.35(m,1H). 1 H NMR (400MHz, CD 3 OD) δ=1.78-1.84(m,3H),1.84-1.87(m,3H),2.04-2.12(m,2H),2.45-2.55(m,2H),3.20- 3.25(m,3H),3.58-3.67(m,3H),3.77-3.81(m,2H),3.81-3.89(m,3H),6.18-6.25(m,1H),6.76-6.83(m,1H ),7.24-7.31(m,1H),7.37-7.42(m,1H),7.50-7.57(m,1H),7.58-7.67(m,1H),8.07-8.11(m,1H),8.29-8.35 (m,1H).
实施例5Example 5
Figure PCTCN2022129958-appb-000052
Figure PCTCN2022129958-appb-000052
第一步first step
将化合物1-2(700.00毫克，3.23毫摩尔，1当量)和化合物G(441.96毫克，2.70毫摩尔,1.17当量)溶于二氧六环(5毫升)，接着向反应液中加入碳酸铯(1.51克，4.63毫摩尔，2当量)，4,5-双(二苯基磷)-9，9-二甲基氧杂蒽(134.04毫克，231.66微摩尔，0.1当量)，三(二亚苄基丙酮)二钯(106.07毫克115.83微摩尔，0.05当量)在100摄氏度氮气保护下反应12小时。反应完成后，加水(10毫升)，乙酸乙酯萃取(10毫升×3)，饱和食盐水(20毫升×3)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1到4：1)纯化得到化合物5-1。Compound 1-2 (700.00 mg, 3.23 mmol, 1 eq) and compound G (441.96 mg, 2.70 mmol, 1.17 eq) were dissolved in dioxane (5 ml), then cesium carbonate ( 1.51 g, 4.63 mmol, 2 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (134.04 mg, 231.66 micromol, 0.1 equiv), tris(diphenylene Dipalladium (106.07 mg, 115.83 micromoles, 0.05 equiv.) was reacted at 100 degrees Celsius under nitrogen protection for 12 hours. After the reaction was complete, add water (10 ml), extract with ethyl acetate (10 ml × 3), wash with saturated brine (20 ml × 3), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate the residue under reduced pressure on silica gel Purified by column chromatography (petroleum ether: ethyl acetate = 100:1 to 4:1) to obtain compound 5-1.
MS-ESI计算值[M+H] +349,实测值349。 MS-ESI calculated [M+H] + 349, found 349.
第二步second step
将化合物5-1(600毫克，1.72毫摩尔，1当量)溶于二氯甲烷(5毫升)，接着向反应液中加入三氟乙酸(196.75毫克，1.72毫摩尔，1当量)，在氮气保护下25摄氏度下反应0.5小时。反应完成后，加入氨水(30毫升)，二氯甲烷萃取(20毫升×3)，有机相经饱和食盐水(30毫升×3)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩得到化合物5-2。Compound 5-1 (600 mg, 1.72 mmol, 1 eq) was dissolved in dichloromethane (5 ml), then trifluoroacetic acid (196.75 mg, 1.72 mmol, 1 eq) was added to the reaction solution, and React at 25°C for 0.5 hours. After the reaction was complete, ammonia water (30 ml) was added, extracted with dichloromethane (20 ml × 3), the organic phase was washed with saturated brine (30 ml × 3), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the compound 5-2.
MS-ESI计算值[M+H] +249,实测值249。 MS-ESI calculated [M+H] + 249, found 249.
第三步third step
将化合物5-2(50.00毫克，201.35微摩尔，1当量)溶于四氢呋喃(2毫升)，接着向反应液中加入中 间体E(95.48毫克，302.03微摩尔，1.5当量)，三(二亚苄基丙酮)二钯(18.44毫克，20.14微摩尔，0.1当量)，2-二叔丁基膦-2′，4′，6′-三异丙基联苯(19.20毫克，40.27微摩尔，0.2当量)，叔丁醇钠(38.70毫克，402.71微摩尔，2当量)，在70摄氏度氮气保护下反应12小时。反应完成后，加水(10毫升)，乙酸乙酯萃取(10毫升×3)，有机相用饱和食盐水(20毫升×3)洗涤，无水硫酸钠干燥，过滤，浓缩，粗品经硅胶薄层色谱板(石油醚：乙酸乙酯＝1：1)纯化，再经高效液相色谱(色谱柱：Waters Xbridge 150*25毫米*5微米；流动相A：氨水，流动相B：乙腈；B％:34％-64％，9分钟)，分离得到化合物5。Compound 5-2 (50.00 mg, 201.35 micromoles, 1 equivalent) was dissolved in tetrahydrofuran (2 ml), and intermediate E (95.48 mg, 302.03 micromoles, 1.5 equivalents), tris(dibenzylidene Acetone) dipalladium (18.44 mg, 20.14 micromoles, 0.1 equivalents), 2-di-tert-butylphosphine-2', 4', 6'-triisopropylbiphenyl (19.20 mg, 40.27 micromoles, 0.2 equivalents ), sodium tert-butoxide (38.70 mg, 402.71 micromoles, 2 equivalents), reacted for 12 hours at 70 degrees Celsius under nitrogen protection. After the reaction was complete, add water (10 ml), extract with ethyl acetate (10 ml × 3), wash the organic phase with saturated brine (20 ml × 3), dry over anhydrous sodium sulfate, filter, concentrate, and the crude product was passed through a thin layer of silica gel Chromatographic plate (petroleum ether: ethyl acetate = 1:1) purification, and then through high performance liquid chromatography (chromatographic column: Waters Xbridge 150*25 millimeters*5 micron; Mobile phase A: ammonia water, mobile phase B: acetonitrile; B% :34%-64%, 9 minutes), isolated compound 5.
MS-ESI计算值[M+H] +528,实测值528。 MS-ESI calculated value [M+H] + 528, found value 528.
1HNMR(400MHz,CD 3OD)δ＝8.39-8.31(m,1H),8.15-8.09(m,1H),7.69-7.60(m,1H),7.60-7.54(m,1H),7.60-7.53(m,1H),7.44-7.438(m,1H),7.35-7.27(m,1H),6.87-6.79(m,1H),6.30-6.24(m,1H),3.72-3.66(m,11H),1.89-1.82(m,10H)。 1 HNMR (400MHz, CD 3 OD) δ=8.39-8.31(m,1H),8.15-8.09(m,1H),7.69-7.60(m,1H),7.60-7.54(m,1H),7.60-7.53 (m,1H),7.44-7.438(m,1H),7.35-7.27(m,1H),6.87-6.79(m,1H),6.30-6.24(m,1H),3.72-3.66(m,11H) ,1.89-1.82(m,10H).
实施例6Example 6
Figure PCTCN2022129958-appb-000053
Figure PCTCN2022129958-appb-000053
第一步first step
将化合物1-2(800.00毫克，2.65毫摩尔，1当量)和化合物H(393.68毫克，3.97毫摩尔，1.5当量)溶于二氧六环(10毫升)，接着向反应液中加入碳酸铯(1.73克，5.30毫摩尔，2当量)，醋酸钯(59.44毫克，264.76微摩尔，0.1当量)，三叔丁基膦四氟硼酸盐(76.81毫克，264.76微摩尔，0.1当量)，在100摄氏度氮气保护下反应12小时。反应完成后，反应液过滤，滤液减压浓缩的残渣经硅胶柱色谱(石油醚：乙酸乙酯＝100：1-10:1)纯化得化合物6-1。Compound 1-2 (800.00 mg, 2.65 mmol, 1 eq) and compound H (393.68 mg, 3.97 mmol, 1.5 eq) were dissolved in dioxane (10 mL), then cesium carbonate ( 1.73 g, 5.30 mmol, 2 equiv), palladium acetate (59.44 mg, 264.76 micromol, 0.1 equiv), tri-tert-butylphosphine tetrafluoroborate (76.81 mg, 264.76 micromol, 0.1 equiv), at 100°C The reaction was carried out for 12 hours under the protection of nitrogen. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-10:1) to obtain compound 6-1.
MS-ESI计算值[M+H] +321,实测值321。 MS-ESI calculated [M+H] + 321, found 321.
第二步second step
将化合物6-1(350毫克，1.09毫摩尔，1当量)溶于二氯甲烷(3毫升)，接着在室温下向反应液中加入三氟乙酸(3毫升)，在25摄氏度下反应0.5小时。反应完成后，加入碳酸氢钠饱和水溶液(30毫升)，二氯甲烷萃取(10毫升×3)，有机相经饱和食盐水(40毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩。粗品未经纯化得到化合物6-2。Compound 6-1 (350 mg, 1.09 mmol, 1 equivalent) was dissolved in dichloromethane (3 ml), then trifluoroacetic acid (3 ml) was added to the reaction solution at room temperature, and reacted at 25 degrees Celsius for 0.5 hours . After the reaction was complete, add saturated aqueous sodium bicarbonate (30 ml), extract with dichloromethane (10 ml × 3), wash the organic phase with saturated brine (40 ml × 2), dry the organic phase with anhydrous sodium sulfate, and filter ,concentrate. The crude product was not purified to obtain compound 6-2.
MS-ESI计算值[M+H] +221,实测值221。 MS-ESI calculated value [M+H] + 221, found value 221.
第三步third step
将化合物6-2(70.00毫克，317.80微摩尔，1当量)溶于二氧六环(3毫升)，接着向反应液中加入中间体E(105.49毫克，333.69微摩尔，1.05当量)，叔丁醇钠(61.08毫克，635.59微摩尔，2当量)，2-二环己基磷-2,4,6-三异丙基联苯(30.30毫克，63.56微摩尔，0.2当量),三(二亚苄基丙酮)二钯(29.10毫克，31.78微摩尔，0.1当量)，在100摄氏度下氮气保护下反应1小时。反应完成后，反应液用硅藻土过滤，滤液减压浓缩的粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)纯化的残渣经高效液相色谱(色谱柱：Phenomenex C18 75*30毫米*3微米；流动相：流动相A：0.5％甲酸水溶液；流动相B：乙腈；B％:25％-55％，7分钟)分离得到化合物6。Compound 6-2 (70.00 mg, 317.80 micromoles, 1 equiv) was dissolved in dioxane (3 ml), then Intermediate E (105.49 mg, 333.69 micromoles, 1.05 equiv), tert-butyl Sodium alkoxide (61.08 mg, 635.59 micromoles, 2 equivalents), 2-dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (30.30 mg, 63.56 micromoles, 0.2 equivalents), tris(dibenzyl Dipalladium (29.10 mg, 31.78 micromole, 0.1 eq) was reacted at 100°C under nitrogen protection for 1 hour. After the reaction was completed, the reaction solution was filtered with diatomaceous earth, and the crude product concentrated under reduced pressure of the filtrate was purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1), and the residue was purified by high performance liquid chromatography (chromatographic column: Phenomenex C18 75 *30 mm*3 micron; mobile phase: mobile phase A: 0.5% aqueous formic acid; mobile phase B: acetonitrile; B%: 25%-55%, 7 minutes) to obtain compound 6.
MS-ESI计算值[M+H] +500,实测值500。 MS-ESI calculated value [M+H] + 500, measured value 500.
1HNMR(400MHz,CD 3OD)δ＝8.39-8.30(m,1H),8.15-8.09(m,1H),7.69-7.61(m,1H),7.60-7.50(m,1H),7.46-7.37(m,1H),7.35-7.25(m,1H),6.86-6.78(m,1H),6.33-6.21(m,1H),4.85(s,4H),4.09-4.02(m,4H),3.72-3.64(m,3H),1.91-1.87(m,3H),1.84(s,3H)。 1 HNMR (400MHz, CD 3 OD) δ=8.39-8.30(m,1H),8.15-8.09(m,1H),7.69-7.61(m,1H),7.60-7.50(m,1H),7.46-7.37 (m,1H),7.35-7.25(m,1H),6.86-6.78(m,1H),6.33-6.21(m,1H),4.85(s,4H),4.09-4.02(m,4H),3.72 -3.64(m,3H),1.91-1.87(m,3H),1.84(s,3H).
实施例7Example 7
Figure PCTCN2022129958-appb-000054
Figure PCTCN2022129958-appb-000054
第一步first step
将化合物7-1(5.00克，31.33毫摩尔，1当量)溶于二氯甲烷(100毫升)，接着向反应液中加入4-二甲氨基吡啶(382.80毫克，3.13毫摩尔，0.1当量)和二碳酸二叔丁酯(17.10克，78.33毫摩尔，18.00毫升，2.5当量)，在25摄氏度下反应12小时。反应完成后，加水(50毫升)稀释，二氯甲烷萃取(20毫升×3)，合并的有机相用饱和食盐水溶液(20毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-5：1)纯化得到化合物7-2。Compound 7-1 (5.00 g, 31.33 mmol, 1 eq) was dissolved in dichloromethane (100 ml), followed by adding 4-dimethylaminopyridine (382.80 mg, 3.13 mmol, 0.1 eq) and Di-tert-butyl dicarbonate (17.10 g, 78.33 mmol, 18.00 ml, 2.5 equivalents) was reacted at 25°C for 12 hours. After completion of the reaction, dilute with water (50 ml), extract with dichloromethane (20 ml × 3), wash the combined organic phase with saturated saline solution (20 ml × 2), dry the organic phase with anhydrous sodium sulfate, filter, and reduce The residue concentrated under reduced pressure was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-5:1) to obtain compound 7-2.
MS-ESI计算值[M+H] +360,实测值360。 MS-ESI calculated value [M+H] + 360, found value 360.
第二步second step
将化合物7-2(500.00毫克，1.39毫摩尔，1当量)溶于乙腈(20毫升)，接着向反应液中加入碳酸钾(576.17毫克，4.17毫摩尔，3当量)和中间体D(249.29毫克，2.08毫摩尔，1.5当量)，在60摄氏度下反应16小时。反应完成后，加水(60毫升)稀释，乙酸乙酯萃取(20毫升×3)，有机相用无水硫酸钠干燥，过滤，减压浓缩。粗品经高效液相色谱(色谱柱：Waters Xbridge 150*25毫米*5微米；流动相：流动相A：氨水溶液；流动相B：乙腈；B％:48％-78％，9分钟)分离得到化合物7-3。Compound 7-2 (500.00 mg, 1.39 mmol, 1 equivalent) was dissolved in acetonitrile (20 ml), then potassium carbonate (576.17 mg, 4.17 mmol, 3 equivalents) and Intermediate D (249.29 mg , 2.08 mmoles, 1.5 equivalents), reacted at 60 degrees Celsius for 16 hours. After the reaction was completed, it was diluted with water (60 mL), extracted with ethyl acetate (20 mL×3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by high performance liquid chromatography (column: Waters Xbridge 150*25 mm*5 microns; mobile phase: mobile phase A: ammonia solution; mobile phase B: acetonitrile; B%: 48%-78%, 9 minutes) to obtain Compound 7-3.
MS-ESI计算值[M+H] +407,实测值407。 MS-ESI calculated [M+H] + 407, found 407.
第三步third step
将化合物7-3(400毫克，984.07微摩尔，1当量)溶于二氯甲烷(5毫升)，接着向反应液中加入三氟乙酸(7.70克，67.53毫摩尔，5毫升，68.62当量)，在25摄氏度下反应0.5小时。反应完成后，加二氯甲烷(20毫升)稀释，将反应液滴入饱和的碳酸氢钠溶液(20毫升)，用二氯甲烷萃取(20毫升×2)，分液，有机相用无水硫酸钠干燥，过滤，减压浓缩，得到化合物7-4。Compound 7-3 (400 mg, 984.07 micromole, 1 eq) was dissolved in dichloromethane (5 mL), then trifluoroacetic acid (7.70 g, 67.53 mmol, 5 mL, 68.62 eq) was added to the reaction solution, React at 25 degrees Celsius for 0.5 hours. After the reaction was complete, add dichloromethane (20 ml) to dilute, add the reaction solution dropwise to saturated sodium bicarbonate solution (20 ml), extract with dichloromethane (20 ml×2), separate the layers, and wash the organic phase with anhydrous Dry over sodium sulfate, filter, and concentrate under reduced pressure to obtain compound 7-4.
MS-ESI计算值[M+H] +207,实测值207。 MS-ESI calculated [M+H] + 207, found 207.
第五步the fifth step
将化合物7-4(100毫克，484.86微摩尔，1当量)溶于四氢呋喃(10毫升)，接着向反应液中加入叔丁醇钠(93.19毫克，969.73微摩尔，2当量)，中间体E(153.28毫克，484.86微摩尔，1当量)，2-二环己基磷-2,4,6-三异丙基联苯(46.23毫克，96.97微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(88.80毫克，96.97微摩尔，0.2当量)，置换三次氮气，并在氮气氛围下在70摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)纯化得到的粗品经高效液相色谱(色谱柱：Waters Xbridge 150*50毫米*3微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％:12％-42％，10分钟)分离得到化合物7。Compound 7-4 (100 mg, 484.86 micromoles, 1 equivalent) was dissolved in tetrahydrofuran (10 ml), then sodium tert-butoxide (93.19 mg, 969.73 micromoles, 2 equivalents) was added to the reaction solution, intermediate E ( 153.28 mg, 484.86 micromoles, 1 equiv), 2-dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (46.23 mg, 96.97 micromoles, 0.2 equiv), tris(dibenzylideneacetone) Dipalladium (88.80 mg, 96.97 micromole, 0.2 equiv.), nitrogen was replaced three times, and reacted at 70° C. for 12 hours under nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1) was purified by high performance liquid chromatography (column: Waters Xbridge 150*50 millimeters*3 microns; mobile phase: mobile phase A: formic acid solution ; Mobile phase B: acetonitrile; B%: 12%-42%, 10 minutes) isolated to obtain compound 7.
MS-ESI计算值[M+H] +486,实测值486。 MS-ESI calculated [M+H] + 486, found 486.
1H NMR(400MHz,CD 3OD)δ＝0.72-0.75(m,4H),1.82-1.84(m,3H),1.85-1.87(m,3H),3.89-3.92(m,3H),4.18-4.20(m,4H),7.16-7.24(m,1H),7.39-7.48(m,1H),7.51-7.58(m,1H),7.97-8.04(m,1H),8.10-8.17(m,1H),8.18-8.23(m,1H),8.28-8.41(m,1H). 1 H NMR (400MHz, CD 3 OD) δ = 0.72-0.75 (m, 4H), 1.82-1.84 (m, 3H), 1.85-1.87 (m, 3H), 3.89-3.92 (m, 3H), 4.18- 4.20(m,4H),7.16-7.24(m,1H),7.39-7.48(m,1H),7.51-7.58(m,1H),7.97-8.04(m,1H),8.10-8.17(m,1H ),8.18-8.23(m,1H),8.28-8.41(m,1H).
实施例8Example 8
Figure PCTCN2022129958-appb-000055
Figure PCTCN2022129958-appb-000055
第一步first step
将化合物7-2(800.00毫克，2.22毫摩尔，1当量)和中间体F(518.86毫克，3.34毫摩尔，1.5当量)溶于乙腈(25毫升)，接着向反应液中加入碳酸钾(921.87毫克，6.67毫摩尔，3当量)，在60摄氏度保护下反应16小时。反应完成后，反应液加水(80毫升)，乙酸乙酯萃取(40毫升×2)，有机相用 无水硫酸钠干燥，过滤。滤液浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝50：1-3：1)纯化得化合物8-1。Compound 7-2 (800.00 mg, 2.22 mmol, 1 eq) and intermediate F (518.86 mg, 3.34 mmol, 1.5 eq) were dissolved in acetonitrile (25 ml), then potassium carbonate (921.87 mg , 6.67 mmoles, 3 equivalents), and reacted for 16 hours under protection at 60 degrees Celsius. After the reaction was complete, the reaction solution was added with water (80 ml), extracted with ethyl acetate (40 ml x 2), and the organic phase was dried over anhydrous sodium sulfate and filtered. The residue from the concentrated filtrate was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1-3:1) to obtain compound 8-1.
MS-ESI计算值[M+H]+443，实测值343，287。MS-ESI calculated value [M+H]+443, found value 343,287.
1HNMR(400MHz,CD 3OD)δ＝7.89(s,1H),4.35-4.13(m,4H),4.06-3.91(m,3H),1.55-1.31(m,18H),0.93-0.89(m,2H)。 1 HNMR (400MHz, CD 3 OD) δ = 7.89(s, 1H), 4.35-4.13(m, 4H), 4.06-3.91(m, 3H), 1.55-1.31(m, 18H), 0.93-0.89(m ,2H).
第二步second step
将化合物8-1(900.00毫克，2.03毫摩尔，1当量)溶于二氯甲烷(8毫升)，接着在室温下向反应液中加入三氟乙酸(12.32克，108.05毫摩尔，8毫升，53.12当量)，在25摄氏度下反应2小时。反应完成后，加入饱和碳酸氢钠水溶液(150毫升)，二氯甲烷萃取(10毫升×3)，有机相经饱和食盐水(30毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩得到化合物8-2。Compound 8-1 (900.00 mg, 2.03 mmol, 1 equivalent) was dissolved in dichloromethane (8 ml), and then trifluoroacetic acid (12.32 g, 108.05 mmol, 8 ml, 53.12 Equivalent), reacted at 25 degrees Celsius for 2 hours. After the reaction was complete, add saturated aqueous sodium bicarbonate (150 ml), extract with dichloromethane (10 ml × 3), wash the organic phase with saturated brine (30 ml × 2), dry the organic phase with anhydrous sodium sulfate, and filter , concentrated to obtain compound 8-2.
MS-ESI计算值[M+H] +243，实测值243。 MS-ESI calculated [M+H] +243 , found 243.
1HNMR(400MHz,CD 3OD)δ＝7.72-7.53(m,1H),4.20-4.13(m,2H),4.06(s,2H),4.01(br s,3H),1.59-1.53(m,2H)。 1 HNMR (400MHz, CD 3 OD) δ=7.72-7.53(m,1H),4.20-4.13(m,2H),4.06(s,2H),4.01(br s,3H),1.59-1.53(m, 2H).
第三步third step
将化合物8-2(150.00毫克，619.26微摩尔，1当量)，中间体E(195.76毫克，619.26微摩尔，1当量)溶于溶于四氢呋喃(10毫升)，接着向反应液中加入叔丁醇钠(119.02毫克，1.24毫摩尔，2当量)，三(二亚苄基丙酮)二钯(56.71毫克，61.93微摩尔，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(59.04毫克，123.85微摩尔，0.2当量)，在微波反应器中90摄氏度下反应1小时。反应完成后，反应液用硅藻土过滤，滤液减压浓缩。粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)纯化的残渣经高效液相色谱(色谱柱：Waters Xbridge 150*25毫米*5微米；流动相：流动相A：氨水：水(V:V)＝10:1的水溶液；流动相B：乙腈；B％:28％-58％，9分钟)分离得到化合物8。Compound 8-2 (150.00 mg, 619.26 micromoles, 1 equivalent), Intermediate E (195.76 mg, 619.26 micromoles, 1 equivalent) were dissolved in tetrahydrofuran (10 ml), and then tert-butanol was added to the reaction solution Sodium (119.02 mg, 1.24 mmol, 2 equiv), tris(dibenzylideneacetone)dipalladium (56.71 mg, 61.93 micromol, 0.1 equiv), 2-dicyclohexylphosphine-2,4,6-triiso Propylbiphenyl (59.04 mg, 123.85 micromole, 0.2 eq) was reacted in a microwave reactor at 90°C for 1 hour. After the reaction was completed, the reaction solution was filtered with diatomaceous earth, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1), and the residue was purified by high-performance liquid chromatography (column: Waters Xbridge 150*25 mm*5 microns; mobile phase: mobile phase A: ammonia water: water (V:V)=10:1 aqueous solution; mobile phase B: acetonitrile; B%: 28%-58%, 9 minutes) isolated to obtain compound 8.
MS-ESI计算值[M+H] +522,实测值522。 MS-ESI calculated value [M+H] + 522, found value 522.
1HNMR(400MHz,CD 3OD)δ＝8.37-8.29(m,1H),8.26-8.22(m,1H),8.05-8.01(m,1H),7.61-7.54(m,1H),7.49-7.44(m,1H),7.27-7.19(m,1H),4.34-4.28(m,2H),4.25-4.19(m,2H),3.95(s,3H),1.90-1.83(m,6H),1.70-1.63(m,2H)。 1 HNMR (400MHz, CD 3 OD) δ=8.37-8.29(m,1H),8.26-8.22(m,1H),8.05-8.01(m,1H),7.61-7.54(m,1H),7.49-7.44 (m,1H),7.27-7.19(m,1H),4.34-4.28(m,2H),4.25-4.19(m,2H),3.95(s,3H),1.90-1.83(m,6H),1.70 -1.63(m,2H).
实施例9Example 9
Figure PCTCN2022129958-appb-000056
Figure PCTCN2022129958-appb-000056
第一步first step
将化合物7-2(600.00毫克，1.67毫摩尔，1当量)和化合物I(243.03毫克，2.50毫摩尔，1.5当量)溶于乙腈(8毫升)，接着向反应液中加入N，N-二异丙基乙胺(646.57毫克，5.00毫摩尔，871.37微升，3当量)，在60摄氏度保护下反应12小时。反应完成后，反应液加水(40毫升)，乙酸乙酯萃取(20毫升×2)，有机相用无水硫酸钠干燥，过滤。滤液浓缩的残渣经高效液相色谱(色谱柱：Waters Xbridge 150*25毫米*5微米；流动相：流动相A：氨水：水(V:V)＝10:1的水溶液；流动相B：乙腈；B％:55％-85％，9分钟)分离纯化得化合物9-1。Compound 7-2 (600.00 mg, 1.67 mmol, 1 eq) and compound I (243.03 mg, 2.50 mmol, 1.5 eq) were dissolved in acetonitrile (8 ml), then N, N-diiso Propylethylamine (646.57 mg, 5.00 mmol, 871.37 microliters, 3 equivalents) was reacted at 60°C for 12 hours. After the reaction was complete, the reaction solution was added with water (40 ml), extracted with ethyl acetate (20 ml×2), and the organic phase was dried over anhydrous sodium sulfate and filtered. The concentrated residue of the filtrate is subjected to high performance liquid chromatography (chromatographic column: Waters Xbridge 150*25 mm*5 microns; mobile phase: mobile phase A: ammoniacal liquor: water (V:V)=10:1 aqueous solution; mobile phase B: acetonitrile ; B%: 55%-85%, 9 minutes) isolated and purified to obtain compound 9-1.
MS-ESI计算值[M+H] +421，实测值421。 MS-ESI calculated [M+H] + 421, found 421.
1HNMR(400MHz,CDCl 3)δ＝7.81(s,1H),4.09(s,4H),3.90(s,3H),2.22(t,J＝7.58Hz,4H),1.88(t,J＝7.52Hz,2H),1.42(s,18H)。 1 HNMR (400MHz, CDCl 3 )δ=7.81(s,1H),4.09(s,4H),3.90(s,3H),2.22(t,J=7.58Hz,4H),1.88(t,J=7.52 Hz,2H), 1.42(s,18H).
第二步second step
将化合物9-1(360.00毫克，856.12微摩尔，1当量)溶于二氯甲烷(4毫升)，接着在室温下向反应液中加入三氟乙酸(9.24克，81.04毫摩尔，6毫升，94.66当量)，在25摄氏度下反应1小时。反应完成后，加入饱和碳酸氢钠水溶液(15毫升)，二氯甲烷萃取(10毫升×2)，有机相经饱和食盐水(30毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩得到化合物9-2。Compound 9-1 (360.00 mg, 856.12 micromole, 1 equivalent) was dissolved in dichloromethane (4 ml), and then trifluoroacetic acid (9.24 g, 81.04 mmol, 6 ml, 94.66 Equivalent), reacted at 25 degrees Celsius for 1 hour. After the reaction was complete, add saturated aqueous sodium bicarbonate (15 ml), extract with dichloromethane (10 ml × 2), wash the organic phase with saturated brine (30 ml × 2), dry the organic phase with anhydrous sodium sulfate, and filter , concentrated to obtain compound 9-2.
MS-ESI计算值[M+H] +221，实测值221。 MS-ESI calculated [M+H] +221 , found 221.
1HNMR(400MHz,CDCl 3)δ＝7.66(s,1H),7.27(s,1H),4.00(s,5H),3.96(s,3H),2.18(d,J＝7.70Hz,3H),2.20-2.17(m,1H),1.88-1.82(m,2H)。 1 HNMR (400MHz, CDCl 3 )δ=7.66(s,1H),7.27(s,1H),4.00(s,5H),3.96(s,3H),2.18(d,J=7.70Hz,3H), 2.20-2.17(m,1H),1.88-1.82(m,2H).
第三步third step
将化合物9-2(120.00毫克，544.79微摩尔，1当量)，中间体E(172.22毫克，544.79微摩尔，1当量)溶于溶于四氢呋喃(10毫升)，接着向反应液中加入叔丁醇钠(104.71毫克，1.09毫摩尔，2当量)，三(二亚苄基丙酮)二钯(49.89毫克，54.48微摩尔，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(51.94毫克，108.96微摩尔，0.2当量)，在氮气保护性70摄氏度下反应12小时。反应完成后，反应液用硅藻土过滤，滤液减压浓缩。粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)纯化的残渣经高效液相色谱(色谱柱：Waters Xbridge 150*25毫米*5微米；流动相：流动相A：氨水：水(V:V)＝10:1的水溶液；流动相B：乙腈；B％:32％-62％，9分钟)分离得到化合物9。Compound 9-2 (120.00 mg, 544.79 micromoles, 1 equivalent), Intermediate E (172.22 mg, 544.79 micromoles, 1 equivalent) were dissolved in tetrahydrofuran (10 ml), and then tert-butanol was added to the reaction solution Sodium (104.71 mg, 1.09 mmol, 2 equiv), tris(dibenzylideneacetone)dipalladium (49.89 mg, 54.48 micromol, 0.1 equiv), 2-dicyclohexylphosphine-2,4,6-triiso Propylbiphenyl (51.94 mg, 108.96 micromoles, 0.2 equivalents) was reacted under nitrogen protection at 70°C for 12 hours. After the reaction was completed, the reaction solution was filtered with diatomaceous earth, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1), and the residue was purified by high-performance liquid chromatography (column: Waters Xbridge 150*25 mm*5 microns; mobile phase: mobile phase A: ammonia water: water (V:V)=10:1 aqueous solution; mobile phase B: acetonitrile; B%: 32%-62%, 9 minutes) isolated to obtain compound 9.
MS-ESI计算值[M+H] +522,实测值522。 MS-ESI calculated value [M+H] + 522, found value 522.
1HNMR(400MHz,CD 3OD)δ＝8.39-8.24(m,1H),8.16-8.08(m,1H),8.04-7.98(m,1H),7.62-7.52(m,1H),7.48-7.38(m,1H),7.24-7.14(m,1H),4.14-4.06(m,4H),3.96-3.91(m,3H),2.33-2.25(m,4H),1.98-1.91(m,2H),1.86(d,J＝13.46Hz,6H)。 1 HNMR (400MHz, CD 3 OD) δ=8.39-8.24(m,1H),8.16-8.08(m,1H),8.04-7.98(m,1H),7.62-7.52(m,1H),7.48-7.38 (m,1H),7.24-7.14(m,1H),4.14-4.06(m,4H),3.96-3.91(m,3H),2.33-2.25(m,4H),1.98-1.91(m,2H) , 1.86 (d, J=13.46Hz, 6H).
实施例10Example 10
Figure PCTCN2022129958-appb-000057
Figure PCTCN2022129958-appb-000057
第一步first step
将化合物7-2(1克，2.78毫摩尔，1.0当量)溶于乙腈(30毫升)，加入碳酸钾(1.15克，8.34毫摩尔，3当量)和化合物H(413.27毫克，4.17毫摩尔，1.5当量)，反应液在60摄氏度下反应16小时。反应完毕后，加入80毫升水淬灭反应，再用乙酸乙酯(40毫升×2)萃取。有机相经无水硫酸钠干燥，过滤，浓缩。粗品经用硅胶柱层析纯化(石油醚：乙酸乙酯＝5:1)得到化合物10-1。Compound 7-2 (1 g, 2.78 mmol, 1.0 eq) was dissolved in acetonitrile (30 ml), potassium carbonate (1.15 g, 8.34 mmol, 3 eq) and compound H (413.27 mg, 4.17 mmol, 1.5 equivalent), the reaction solution was reacted at 60 degrees Celsius for 16 hours. After the reaction was completed, 80 mL of water was added to quench the reaction, and then extracted with ethyl acetate (40 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 10-1.
MS-ESI计算值[M+H] +423,实测值423。 MS-ESI calculated [M+H] + 423, found 423.
第二步second step
将化合物10-1(910毫克，2.15毫摩尔，1.0当量)溶于二氯甲烷(4毫升)，加入三氟乙酸(9.24克，81.04毫摩尔，37.62当量)，反应液室温反应1小时。反应完毕后，反应液浓缩后加入二氯甲烷(50毫升)稀释，饱和食盐水(50毫升)洗涤。有机相经无水硫酸钠干燥，过滤，浓缩。粗品经用硅胶薄层色谱板纯化(二氯甲烷：甲醇＝8:1)得到化合物10-2。Compound 10-1 (910 mg, 2.15 mmol, 1.0 eq) was dissolved in dichloromethane (4 ml), trifluoroacetic acid (9.24 g, 81.04 mmol, 37.62 eq) was added, and the reaction solution was reacted at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated, diluted with dichloromethane (50 mL), and washed with saturated brine (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel thin-layer chromatography (dichloromethane:methanol=8:1) to obtain compound 10-2.
MS-ESI计算值[M+H] +223,实测值223。 MS-ESI calculated value [M+H] + 223, found value 223.
第三步third step
将化合物10-2(185毫克，832.42微摩尔，1.0当量)，中间体E(263.15毫克，832.42微摩尔，1.0当量)，叔丁醇钠(160.00毫克，1.66毫摩尔，2.0当量)，2-二环己基磷-2,4,6-三异丙基联苯(79.37毫克，166.48微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(76.23毫克，83.24微摩尔，0.1当量)加入四氢呋喃(4毫升)中，氮气置换保护，反应液加热到70摄氏下反应12小时。反应完成后，过滤，滤液减压浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝10:1)和高效液相色谱(色谱柱：Unisil 3-100C18Ultra 150*50毫米*3微米；流动相：流动相A：体积分数0.225％甲酸水溶液；流动相B：乙腈；B％:10％-40％,10分钟)纯化得到化合物10。Compound 10-2 (185 mg, 832.42 micromoles, 1.0 equivalents), Intermediate E (263.15 mg, 832.42 micromoles, 1.0 equivalents), sodium tert-butoxide (160.00 mg, 1.66 mmoles, 2.0 equivalents), 2- Dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (79.37 mg, 166.48 micromoles, 0.2 equiv), tris(dibenzylideneacetone)dipalladium (76.23 mg, 83.24 micromoles, 0.1 equiv) Tetrahydrofuran (4 ml) was added, and nitrogen replacement protection was carried out, and the reaction solution was heated to 70° C. for 12 hours. After the reaction was completed, filter, and the residue of the filtrate concentrated under reduced pressure was subjected to silica gel thin layer chromatography (dichloromethane: methanol=10:1) and high performance liquid chromatography (column: Unisil 3-100C18Ultra 150*50 millimeters*3 microns; Mobile phase: mobile phase A: 0.225% formic acid aqueous solution by volume fraction; mobile phase B: acetonitrile; B%: 10%-40%, 10 minutes) purification to obtain compound 10.
MS-ESI计算值[M+H] +502,实测值502。 MS-ESI calculated value [M+H] + 502, found value 502.
1H NMR(400MHz,CD 3OD)δ＝8.41-8.31(m,1H),8.19(s,1H),8.02(s,1H),7.62-7.52(m,1H),7.49-7.39(m,1H),7.25-7.17(m,1H),4.89(m,4H),4.30(s,4H),3.94(s,3H),1.88(s,3H),1.85(s,3H)。 1 H NMR (400MHz, CD 3 OD) δ=8.41-8.31(m,1H),8.19(s,1H),8.02(s,1H),7.62-7.52(m,1H),7.49-7.39(m, 1H), 7.25-7.17(m,1H), 4.89(m,4H), 4.30(s,4H), 3.94(s,3H), 1.88(s,3H), 1.85(s,3H).
实施例11Example 11
Figure PCTCN2022129958-appb-000058
Figure PCTCN2022129958-appb-000058
第一步first step
将化合物11-1(5.00克，24.51毫摩尔，1当量)溶于二氯甲烷(50毫升)，接着向反应液中加入4-二甲氨基吡啶(299.4毫克，2.45毫摩尔，0.1当量)和二碳酸二叔丁酯(13.37克，61.27毫摩尔，2.5当量)，在25摄氏度下反应1小时。反应完成后，加水(20毫升)稀释，二氯甲烷萃取(10毫升×3)，合并的有机相用饱和食盐水溶液(20毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-20：1)纯化得到化合物11-2。Compound 11-1 (5.00 g, 24.51 mmol, 1 eq) was dissolved in dichloromethane (50 ml), followed by adding 4-dimethylaminopyridine (299.4 mg, 2.45 mmol, 0.1 eq) and Di-tert-butyl dicarbonate (13.37 g, 61.27 mmol, 2.5 equivalents) was reacted at 25°C for 1 hour. After completion of the reaction, dilute with water (20 ml), extract with dichloromethane (10 ml × 3), wash the combined organic phase with saturated saline solution (20 ml × 2), dry the organic phase with anhydrous sodium sulfate, filter, and reduce The residue concentrated under reduced pressure was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-20:1) to obtain compound 11-2.
MS-ESI计算值[M+H] +404,实测值404。 MS-ESI calculated value [M+H] + 404, found value 404.
第二步second step
将化合物11-2(2.00克，4.95毫摩尔，1当量)和中间体F(923.61毫克，5.94毫摩尔，1.2当量)溶于二氧六环(20毫升)，接着向反应液中加入碳酸铯(4.84毫克，14.84毫摩尔，3当量)，醋酸钯(222.14毫克，989.47微摩尔，0.2当量)，(±)-2,2-双(二苯膦基)-1,1-联萘(616.12毫克，989.47微摩尔，0.2当量)，在100摄氏度氮气保护下反应12小时。反应完成后，反应液过滤，滤液浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝50：1-10：1)纯化得化合物11-3。Compound 11-2 (2.00 g, 4.95 mmol, 1 eq) and intermediate F (923.61 mg, 5.94 mmol, 1.2 eq) were dissolved in dioxane (20 ml), then cesium carbonate was added to the reaction solution (4.84 mg, 14.84 mmol, 3 equivalents), palladium acetate (222.14 mg, 989.47 micromoles, 0.2 equivalents), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (616.12 mg, 989.47 micromoles, 0.2 equivalents), reacted for 12 hours at 100 degrees Celsius under nitrogen protection. After the reaction was completed, the reaction solution was filtered, and the concentrated residue of the filtrate was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1-10:1) to obtain compound 11-3.
MS-ESI计算值[M+H] +443，287，实测值443，287。 MS-ESI calculated [M+H] + 443,287, found 443,287.
第三步third step
将化合物11-3(1.00克，2.26毫摩尔，1当量)溶于二氯甲烷(12毫升)，接着在室温下向反应液中加入三氟乙酸(6.16克，54.02毫摩尔，4毫升，23.90当量)，在25摄氏度下反应0.5小时。反应完成后，加入饱和碳酸氢钠水溶液(30毫升)，二氯甲烷萃取(10毫升×3)，有机相经饱和食盐水(40毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩。粗品经高效液相色谱(色谱柱:Waters Xbridge 150*50毫米*10微米；流动相：流动相A：碳酸氢铵水溶液；流动相B：乙腈；B％:24％-54％,10分钟)纯化得到化合物11-4。Compound 11-3 (1.00 g, 2.26 mmol, 1 eq) was dissolved in dichloromethane (12 ml), then trifluoroacetic acid (6.16 g, 54.02 mmol, 4 ml, 23.90 Equivalent), reacted at 25 degrees Celsius for 0.5 hours. After the reaction was complete, add saturated aqueous sodium bicarbonate (30 ml), extract with dichloromethane (10 ml × 3), wash the organic phase with saturated brine (40 ml × 2), dry the organic phase with anhydrous sodium sulfate, and filter ,concentrate. The crude product was subjected to high performance liquid chromatography (column: Waters Xbridge 150*50 mm*10 microns; mobile phase: mobile phase A: aqueous ammonium bicarbonate; mobile phase B: acetonitrile; B%: 24%-54%, 10 minutes) Purification afforded compound 11-4.
MS-ESI计算值[M+H] +243,实测值243。 MS-ESI calculated [M+H] + 243, found 243.
第四步the fourth step
将化合物11-4(138.42毫克，571.47微摩尔，1当量)溶于二氧六环(10毫升)，接着向反应液中加入中间体E(216.78毫克，685.76微摩尔，1.2当量)，碳酸铯(558.58毫克，1.71毫摩尔，3当量)， (±)-2,2-双(二苯膦基)-1,1-联萘(71.17毫克，114.29微摩尔，0.2当量),醋酸钯(25.66毫克，114.29微摩尔，0.2当量)，在100摄氏度下氮气保护下反应12小时。反应完成后，反应液用硅藻土过滤，并用乙酸乙酯洗涤滤饼，滤液用无水硫酸钠干燥，过滤，减压浓缩。粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)纯化的残渣经高效液相色谱(色谱柱：Waters Xbridge 150*25毫米*5微米；流动相：流动相A：氨水：水(V:V)＝10:1的水溶液；流动相B：乙腈；B％:30％-60％，9分钟)分离得到化合物11。Compound 11-4 (138.42 mg, 571.47 micromoles, 1 equiv) was dissolved in dioxane (10 ml), then Intermediate E (216.78 mg, 685.76 micromoles, 1.2 equiv), cesium carbonate (558.58 mg, 1.71 mmol, 3 equivalents), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (71.17 mg, 114.29 micromoles, 0.2 equivalents), palladium acetate (25.66 mg, 114.29 micromoles, 0.2 equivalents), reacted under nitrogen protection at 100 degrees Celsius for 12 hours. After the reaction was completed, the reaction solution was filtered with diatomaceous earth, and the filter cake was washed with ethyl acetate. The filtrate was dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1), and the residue was purified by high-performance liquid chromatography (column: Waters Xbridge 150*25 mm*5 microns; mobile phase: mobile phase A: ammonia water: water (V:V)=10:1 aqueous solution; mobile phase B: acetonitrile; B%: 30%-60%, 9 minutes) isolated to obtain compound 11.
MS-ESI计算值[M+H] +521.9,实测值521.9。 MS-ESI calculated [M+H] + 521.9, found 521.9.
1HNMR(400MHz,CD 3OD)δ＝8.49-8.37(m,1H),8.09-8.02(m,1H),7.62-7.51(m,1H),7.48-7.39(m,1H),7.30-7.24(m,1H),7.22-7.14(m,1H),4.34-4.27(m,2H),4.24-4.19(m,2H),3.94-3.90(m,3H),1.90-1.87(m,3H),1.86-1.83(m,3H),1.72-1.64(m,2H)。 1 HNMR (400MHz, CD 3 OD) δ=8.49-8.37(m,1H),8.09-8.02(m,1H),7.62-7.51(m,1H),7.48-7.39(m,1H),7.30-7.24 (m,1H),7.22-7.14(m,1H),4.34-4.27(m,2H),4.24-4.19(m,2H),3.94-3.90(m,3H),1.90-1.87(m,3H) ,1.86-1.83(m,3H),1.72-1.64(m,2H).
实施例12Example 12
Figure PCTCN2022129958-appb-000059
Figure PCTCN2022129958-appb-000059
第一步first step
将化合物11-2(1.00，2.47毫摩尔，1当量)溶于二氧六环(20毫升)中，接着在向反应液中加入中间体D(411.28毫克，3.44毫摩尔，1.39当量)，1,1’-联萘-2，2’-双二苯膦(308.06毫克，494.74微摩尔,0.2当量)，碳酸铯(1.61克，4.95毫摩尔，2当量)，醋酸钯(111.07毫克，494.74微摩尔0.2当量)在100摄氏度氮气保护下反应12小时。反应完成后，加入水(60毫升)，乙酸乙酯萃取(60毫升×3)，有机相经饱和食盐水(60毫升×3)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩。粗品经用硅胶薄层色谱板纯化(二氯甲烷：甲醇＝10：1)得到化合物12-1。Compound 11-2 (1.00, 2.47 mmol, 1 eq) was dissolved in dioxane (20 ml), then Intermediate D (411.28 mg, 3.44 mmol, 1.39 eq) was added to the reaction solution, 1 , 1'-binaphthyl-2,2'-bisdiphenylphosphine (308.06 mg, 494.74 μmol, 0.2 eq), cesium carbonate (1.61 g, 4.95 mmol, 2 eq), palladium acetate (111.07 mg, 494.74 μm Mole 0.2 equivalent) was reacted for 12 hours at 100 degrees Celsius under the protection of nitrogen. After the reaction was complete, water (60 mL) was added, extracted with ethyl acetate (60 mL×3), the organic phase was washed with saturated brine (60 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1) to obtain compound 12-1.
MS-ESI计算值[M-156+H] +251,实测值251。 MS-ESI calculated value [M-156+H] + 251, measured value 251.
第二步second step
将化合物12-1(250.00毫克，615.04微摩尔，1当量)溶于二氯甲烷(2毫升)，接着向反应液中加入三氟乙酸(350.64毫克,227.69微升，3.08毫摩尔，5当量)，在25摄氏度下反应0.5小时。反应完成后，加水(100毫升)，乙酸乙酯萃取(60毫升×3)，有机相用饱和食盐水(60毫升×3)洗涤，无水硫酸钠干燥，过滤，浓缩得到化合物12-2。Compound 12-1 (250.00 mg, 615.04 μmol, 1 eq) was dissolved in dichloromethane (2 ml), and then trifluoroacetic acid (350.64 mg, 227.69 μl, 3.08 mmol, 5 eq) was added to the reaction solution , reacted at 25 degrees Celsius for 0.5 hours. After the reaction was complete, water (100 mL) was added, extracted with ethyl acetate (60 mL×3), the organic phase was washed with saturated brine (60 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 12-2.
MS-ESI计算值[M+H] +207,实测值207。 MS-ESI calculated [M+H] + 207, found 207.
1H NMR(400MHz,CD 3OD)δ＝6.80-6.71(m,1H),6.81-6.69(m,1H),4.96-4.76(m,39H),3.98- 3.93(m,3H)0.66(s,2H)。 1 H NMR (400MHz, CD 3 OD) δ=6.80-6.71(m,1H),6.81-6.69(m,1H),4.96-4.76(m,39H),3.98-3.93(m,3H)0.66(s ,2H).
第三步third step
将化合物12-2(80毫克，387.89微摩尔，1当量)溶于四氢呋喃(5毫升)，接着向反应液中加入中间体E(122.62毫克，387.89微摩尔，1当量)、2-二叔丁基膦-2′,4′,6′-三异丙基联苯(36.98毫克,77.58微摩尔,0.2当量)，三(二亚苄基丙酮)二钯(71.04毫克，77.58微摩尔，0.2当量)、叔丁醇钠(74.56毫克，775.78微摩尔，2当量)在70摄氏度氮气保护下反应12小时。反应完成后，加水(100毫升)，乙酸乙酯萃取(60毫升×3)，有机相用饱和食盐水(60毫升×3)洗涤，无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝20：1到4：1)纯化得到粗品，粗品经高效液相色谱(色谱柱：Unisil 3-100C18Ultra 150*50毫米*3微米；流动相A：甲酸水溶液，流动相B：乙腈；B％:20％-50％，10分钟)分离得到化合物12。Compound 12-2 (80 mg, 387.89 micromoles, 1 equivalent) was dissolved in tetrahydrofuran (5 milliliters), then Intermediate E (122.62 mg, 387.89 micromoles, 1 equivalent), 2-di-tert-butyl Phosphine-2′,4′,6′-triisopropylbiphenyl (36.98 mg, 77.58 micromole, 0.2 equiv), tris(dibenzylideneacetone) dipalladium (71.04 mg, 77.58 micromole, 0.2 equiv ), sodium tert-butoxide (74.56 mg, 775.78 micromoles, 2 equivalents) were reacted for 12 hours at 70 degrees Celsius under nitrogen protection. After the reaction was complete, add water (100 ml), extract with ethyl acetate (60 ml × 3), wash the organic phase with saturated brine (60 ml × 3), dry over anhydrous sodium sulfate, filter, and concentrate the residue under reduced pressure on silica gel The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 20:1 to 4:1), and the crude product was subjected to high performance liquid chromatography (column: Unisil 3-100C18Ultra 150*50mm*3 microns; mobile phase A: formic acid Aqueous solution, mobile phase B: acetonitrile; B%: 20%-50%, 10 minutes) isolated to obtain compound 12.
MS-ESI计算值[M+H] +486,实测值486。 MS-ESI calculated [M+H] + 486, found 486.
1H NMR(400MHz,CD 3OD)δ＝8.34-8.40(m,1H),8.03-7.99(m,1H),7.56-7.48(m,1H),7.42-7.36(m,1H),7.20-7.14(m,2H),4.19-4.16(m,4H),3.90-3.86(m,3H),1.87-1.82(m,6H),0.77-0.74(m,4H)。 1 H NMR (400MHz, CD 3 OD) δ=8.34-8.40(m,1H),8.03-7.99(m,1H),7.56-7.48(m,1H),7.42-7.36(m,1H),7.20- 7.14 (m, 2H), 4.19-4.16 (m, 4H), 3.90-3.86 (m, 3H), 1.87-1.82 (m, 6H), 0.77-0.74 (m, 4H).
实施例13Example 13
Figure PCTCN2022129958-appb-000060
Figure PCTCN2022129958-appb-000060
第一步first step
将化合物11-2(1克，2.47毫摩尔,1当量)溶于二氧六环(20毫升)，接着向反应液中加入化合物I(396.64毫克，2.97毫摩尔，1.2当量)，碳酸铯(2.42克，7.42毫摩尔，3当量)，[1,1-双(二苯基膦)二茂铁]二氯化钯二氯甲烷(202.01毫克，247.37毫摩尔，0.1当量)在100摄氏度氮气保护下反应2小时。反应完成后，用硅藻土过滤，滤液倒入少量冰水用二氯甲烷(30毫升×3)萃取，饱和食盐水洗涤2次，最后用无水硫酸钠干燥，过滤，滤液减压浓缩，经层析柱(石油醚/乙酸乙酯＝3/1)纯化得到化合物13-1。Compound 11-2 (1 g, 2.47 mmol, 1 eq) was dissolved in dioxane (20 ml), then compound I (396.64 mg, 2.97 mmol, 1.2 eq), cesium carbonate ( 2.42 g, 7.42 mmol, 3 eq), [1,1-bis(diphenylphosphine) ferrocene] dichloropalladium dichloromethane (202.01 mg, 247.37 mmol, 0.1 eq) at 100°C under nitrogen protection The reaction was carried out for 2 hours. After the reaction was completed, filter with diatomaceous earth, pour the filtrate into a small amount of ice water and extract with dichloromethane (30 ml × 3), wash with saturated brine twice, and finally dry with anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure. Compound 13-1 was obtained by column chromatography (petroleum ether/ethyl acetate=3/1).
MS-ESI计算值[M-156+H] +265,实测值265。 MS-ESI calculated value [M-156+H] + 265, found value 265.
第二步second step
将化合物13-1(360毫克，856.12微摩尔，1当量)溶于二氯甲烷(8毫升)，接着向反应液中加入三氟乙酸(8毫升)，在25摄氏度下反应0.5小时。反应完成后，倒入少量冰水中，用碳酸钠调节pH＝8，用二氯甲烷(30毫升×2)萃取，有机相经饱和食盐水(20毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤， 浓缩。粗品经硅胶薄层色谱板(二氯甲烷/甲醇＝10/1)纯化得到化合物13-2。Compound 13-1 (360 mg, 856.12 micromole, 1 eq) was dissolved in dichloromethane (8 ml), then trifluoroacetic acid (8 ml) was added to the reaction solution, and reacted at 25°C for 0.5 hour. After the reaction is complete, pour into a small amount of ice water, adjust the pH=8 with sodium carbonate, extract with dichloromethane (30 ml × 2), wash the organic phase with saturated brine (20 ml × 2), and wash the organic phase with anhydrous sulfuric acid Dry over sodium, filter and concentrate. The crude product was purified by silica gel thin-layer chromatography (dichloromethane/methanol=10/1) to obtain compound 13-2.
MS-ESI计算值[M+H] +221,实测值221。 MS-ESI calculated value [M+H] + 221, found value 221.
第三步third step
将化合物13-2(70毫克，317.79微摩尔，1当量)，中间体E(110.51毫克，349.57微摩尔，1.1当量)溶于无水四氢呋喃(10毫升)，接着向反应液中加入叔丁醇钠(91.62毫克，953.37微摩尔，3当量)，2-二-叔丁基膦-2’,4’,6’-三异丙基联苯(30.30毫克，63.56微摩尔，0.2当量)，醋酸钯(43.65毫克，47.67微摩尔，0.15当量)，在氮气保护下70摄氏度下反应8小时。反应完成后，反应液倒入少量冰水用乙酸乙酯(30毫升×3)萃取，饱和食盐水洗涤2次，最后用无水硫酸钠干燥，过滤，滤液减压浓缩浓缩，经高效液相色谱(色谱柱Phenomenex Synergi C18 150*25毫米*10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：26％-56％，10分钟)分离纯化得到化合物13。Compound 13-2 (70 mg, 317.79 micromoles, 1 equiv), Intermediate E (110.51 mg, 349.57 micromoles, 1.1 equiv) were dissolved in anhydrous tetrahydrofuran (10 ml), and then tert-butanol was added to the reaction solution Sodium (91.62 mg, 953.37 micromoles, 3 equiv), 2-di-tert-butylphosphine-2',4',6'-triisopropylbiphenyl (30.30 mg, 63.56 micromoles, 0.2 equiv), acetic acid Palladium (43.65 mg, 47.67 micromole, 0.15 eq.) was reacted at 70° C. for 8 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into a small amount of ice water and extracted with ethyl acetate (30 ml × 3), washed twice with saturated brine, and finally dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and subjected to high-efficiency liquid phase Chromatography (column Phenomenex Synergi C18 150*25 mm*10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 26%-56%, 10 minutes) separation and purification to obtain compound 13.
MS-ESI计算值[M+H] +500,实测值500。 MS-ESI calculated value [M+H] + 500, measured value 500.
1H NMR(400MHz,CD 3OD)δppm 1.75-1.87(m,6H)1.89-1.97(m,2H)2.22-2.33(m,4H)3.81-3.94(m,3H)3.99-4.09(m,4H)7.05-7.20(m,2H)7.31-7.42(m,1H)7.44-7.59(m,1H)7.91-8.07(m,1H)8.30-8.43(m,1H). 1 H NMR (400MHz, CD 3 OD) δppm 1.75-1.87(m,6H)1.89-1.97(m,2H)2.22-2.33(m,4H)3.81-3.94(m,3H)3.99-4.09(m,4H )7.05-7.20(m,2H)7.31-7.42(m,1H)7.44-7.59(m,1H)7.91-8.07(m,1H)8.30-8.43(m,1H).
实施例14Example 14
Figure PCTCN2022129958-appb-000061
Figure PCTCN2022129958-appb-000061
第一步first step
将化合物11-2(3克，7.42毫摩尔，1当量)溶于二氧六环(50毫升)，接着向反应液中加入碳酸铯(7.25毫克，22.26毫摩尔，3当量)和化合物H(1.10克，11.13毫摩尔，1.5当量)，三叔丁基膦四氟硼酸盐(430.61毫克，1.48毫摩尔，0.2当量)和醋酸钯(333.22毫克，1.48毫摩尔，0.2当量)。置换三次氮气，并在氮气氛围下在100摄氏度下反应12小时。反应完成后，过滤，减压浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1-1：1)纯化得到化合物14-1。Compound 11-2 (3 g, 7.42 mmol, 1 eq) was dissolved in dioxane (50 ml), then cesium carbonate (7.25 mg, 22.26 mmol, 3 eq) and compound H ( 1.10 g, 11.13 mmol, 1.5 equiv), tri-tert-butylphosphine tetrafluoroborate (430.61 mg, 1.48 mmol, 0.2 equiv) and palladium acetate (333.22 mg, 1.48 mmol, 0.2 equiv). Nitrogen was replaced three times, and reacted at 100°C for 12 hours under a nitrogen atmosphere. After the reaction was completed, it was filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1-1:1) to obtain compound 14-1.
第二步second step
将化合物14-1(1克，2.37毫摩尔，1当量)溶于二氯甲烷(10毫升)，接着向反应液中加入三氟乙酸(15.40克，135.06毫摩尔，10毫升，57.06当量)，在25摄氏度下反应0.5小时。反应完成后，加二氯甲烷(20毫升)稀释，将反应液滴入饱和的碳酸氢钠溶液(20毫升)，用二氯甲烷萃取(20毫升×2)，分液，有 机相用无水硫酸钠干燥，过滤，减压浓缩，得到粗品化合物14-2。Compound 14-1 (1 g, 2.37 mmol, 1 eq) was dissolved in dichloromethane (10 mL), then trifluoroacetic acid (15.40 g, 135.06 mmol, 10 mL, 57.06 eq) was added to the reaction solution, React at 25 degrees Celsius for 0.5 hours. After the reaction was complete, add dichloromethane (20 ml) to dilute, add the reaction solution dropwise to saturated sodium bicarbonate solution (20 ml), extract with dichloromethane (20 ml×2), separate the layers, and wash the organic phase with anhydrous Dry over sodium sulfate, filter, and concentrate under reduced pressure to obtain crude compound 14-2.
第三步third step
将化合物14-2(220毫克，1.65毫摩尔，1当量)溶于四氢呋喃(20毫升)，接着向反应液中加入叔丁醇钠(93.19毫克，969.73微摩尔，2当量)，中间体E(153.28毫克，484.86微摩尔，1当量)，2-二环己基磷-2,4,6-三异丙基联苯(78.65毫克，164.98微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(151.08毫克，164.98微摩尔，0.2当量)，置换三次氮气，并在氮气氛围下在70摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶层析柱(石油醚：乙酸乙酯＝50/1～二氯甲烷：甲醇＝3：1)纯化得到的粗品经高效液相色谱(色谱柱：Waters Xbridge 150*25毫米*10微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％:12％-42％，10分钟)分离得到化合物14。Compound 14-2 (220 mg, 1.65 mmol, 1 eq) was dissolved in tetrahydrofuran (20 ml), then sodium tert-butoxide (93.19 mg, 969.73 micromol, 2 eq) was added to the reaction solution, Intermediate E ( 153.28 mg, 484.86 micromoles, 1 equiv), 2-dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (78.65 mg, 164.98 micromoles, 0.2 equiv), tris(dibenzylideneacetone) Dipalladium (151.08 mg, 164.98 micromole, 0.2 eq.) was replaced with nitrogen three times, and reacted at 70° C. for 12 hours under nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by silica gel chromatography column (petroleum ether: ethyl acetate=50/1～dichloromethane:methanol=3:1), and the crude product obtained by high performance liquid chromatography (chromatographic column: Waters Xbridge 150*25 mm*10 microns ; Mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 12%-42%, 10 minutes) isolated to obtain compound 14.
MS-ESI计算值[M+H] +502,实测值502. MS-ESI calculated value [M+H] + 502, measured value 502.
1H NMR(400MHz,CD 3OD)δ＝1.84(d,J＝13.38Hz,6H),3.86-3.91(m,3H),4.24-4.27(m,4H),4.88(s,4H)7.13-7.21(m,2H),7.36-7.43(m,1H),7.47-7.56(m,1H),7.98-8.05(m,1H),8.37-8.44(m,1H). 1 H NMR (400MHz, CD 3 OD) δ = 1.84 (d, J = 13.38Hz, 6H), 3.86-3.91 (m, 3H), 4.24-4.27 (m, 4H), 4.88 (s, 4H) 7.13- 7.21(m,2H),7.36-7.43(m,1H),7.47-7.56(m,1H),7.98-8.05(m,1H),8.37-8.44(m,1H).
实施例15Example 15
Figure PCTCN2022129958-appb-000062
Figure PCTCN2022129958-appb-000062
第一步first step
将化合物15-1(13.5克，64.77毫摩尔，1当量)，溶于甲醇(200毫升)，接着在0摄氏度下向反应液中逐步添加甲醇钠(14克，259.09毫摩尔，4当量)，在25摄氏度下搅拌12小时。反应完毕后减压浓缩，残液加水(100毫升)稀释，乙酸乙酯萃取(100毫升×3)，有机相用饱和食盐水(100毫升×2)洗涤，无水硫酸钠干燥，过滤，减压浓缩后得到粗产品直接用于下步反应，化合物15-2。Compound 15-1 (13.5 g, 64.77 mmol, 1 eq) was dissolved in methanol (200 ml), then sodium methoxide (14 g, 259.09 mmol, 4 eq) was gradually added to the reaction solution at 0°C, Stir for 12 hours at 25°C. After the reaction was completed, it was concentrated under reduced pressure, the residue was diluted with water (100 ml), extracted with ethyl acetate (100 ml × 3), the organic phase was washed with saturated brine (100 ml × 2), dried over anhydrous sodium sulfate, filtered, and reduced After concentrated under pressure, the crude product was directly used in the next reaction, compound 15-2.
MS-ESI计算值[M+H] +207,实测值207. MS-ESI calculated value [M+H] + 207, measured value 207.
第二步second step
将化合物15-2(5克，31.33毫摩尔，1当量)，15-3(7.15克，62.67毫摩尔，2当量)溶于甲苯(150毫升)，接着向反应液中添加对甲苯磺酸(539.57毫克，3.13毫摩尔，0.1当量)，在氮气保护下130摄氏度下搅拌12小时。反应完毕后，过滤滤液在减压下浓缩浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100:1～2:1)分离纯化得到化合物15-4。Compound 15-2 (5 g, 31.33 mmol, 1 eq), 15-3 (7.15 g, 62.67 mmol, 2 eq) were dissolved in toluene (150 ml), and p-toluenesulfonic acid ( 539.57 mg, 3.13 mmol, 0.1 eq), stirred at 130°C for 12 hours under nitrogen protection. After the reaction was completed, the filtered filtrate was concentrated under reduced pressure, and the concentrated residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-2:1) to obtain compound 15-4.
MS-ESI计算值[M+H]+238，实测值238。MS-ESI calculated value [M+H]+238, found value 238.
1H NMR(400MHz,CDCl 3)δ＝7.12(s,1H)6.00–5.92(m,2H)3.95(s,3H)2.04-2.02(m,6H)。 1 H NMR (400 MHz, CDCl 3 ) δ = 7.12 (s, 1H) 6.00 - 5.92 (m, 2H) 3.95 (s, 3H) 2.04 - 2.02 (m, 6H).
第三步third step
将化合物15-4(1克，4.21毫摩尔，1当量)，D(699.51毫克，5.85毫摩尔，1.39当量)，N,N-二异丙基乙胺(2.18克，16.83毫摩尔，4当量)溶于乙二醇(15毫升)，接着在140摄氏度微波下反应1.5小时。反应完毕后，向反应液中加入水(100毫升)乙酸乙酯萃取(100毫升×2)，有机相用饱和的食盐水(200毫升)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100:1-2:1)分离纯化得到化合物15-5。Compound 15-4 (1 g, 4.21 mmol, 1 eq), D (699.51 mg, 5.85 mmol, 1.39 eq), N,N-diisopropylethylamine (2.18 g, 16.83 mmol, 4 eq ) was dissolved in ethylene glycol (15 ml), followed by reaction under microwave at 140° C. for 1.5 hours. After the reaction was completed, water (100 ml) was added to the reaction solution to extract with ethyl acetate (100 ml×2), the organic phase was washed with saturated brine (200 ml), dried over anhydrous sodium sulfate, filtered, and the concentrated residue was washed with Compound 15-5 was obtained by separation and purification by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-2:1).
MS-ESI计算值[M+H] +285，实测值285。 MS-ESI calculated [M+H] +285 , found 285.
1H NMR(400MHz,CDCl 3)δ＝5.98-5.94(m,1H)5.93-5.88(m,2H)4.29(s,4H)3.87-3.81(m,3H)2.02(s,6H)0.80-0.73(m,4H)。 1 H NMR (400MHz, CDCl 3 )δ=5.98-5.94(m,1H)5.93-5.88(m,2H)4.29(s,4H)3.87-3.81(m,3H)2.02(s,6H)0.80-0.73 (m,4H).
第四步the fourth step
将化合物15-5(442毫克，1.55毫摩尔，1当量)，盐酸羟胺(1.08克，15.54毫摩尔，10当量)，氢氧化钾(174.43毫克，3.11毫摩尔，2当量)溶于乙醇(8毫升)/水(4毫升)中，接着在120摄氏度微波下反应1.5小时反应完毕后，向反应液中加入水(50毫升)，乙酸乙酯萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝10:1)分离纯化得到化合物15-6。Compound 15-5 (442 mg, 1.55 mmol, 1 equivalent), hydroxylamine hydrochloride (1.08 grams, 15.54 mmol, 10 equivalents), potassium hydroxide (174.43 mg, 3.11 mmol, 2 equivalents) were dissolved in ethanol (8 ml)/water (4 ml), followed by reaction at 120°C for 1.5 hours under microwave. Washed with brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the concentrated residue was separated and purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1) to obtain compound 15-6.
MS-ESI计算值[M+H] +207，实测值207。 MS-ESI calculated [M+H] +207 , found 207.
第五步the fifth step
将化合物15-6(30毫克，135.28微摩尔，1当量)，中间体E(42.76毫克，135.28微摩尔，1当量)，叔丁醇钠(39毫克，405.83微摩尔，3当量)，2-二环己基磷-2,4,6-三异丙基联苯(12.9毫克，27.06微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(12.39毫克，13.53微摩尔，0.1当量)溶于二氧六环(2毫升)中，接着在120摄氏度微波下反应1.5小时，反应完毕后，向反应液中加入水(5毫升)乙酸乙酯萃取(5毫升×2)，有机相用饱和的食盐水(10毫升)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝10:1)和高效液相色谱法(色谱柱：Phenomenex Synergi C18 150*25毫米*10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：7％-37％，10分钟)分离纯化得到化合物15。Compound 15-6 (30 mg, 135.28 micromoles, 1 equivalent), Intermediate E (42.76 mg, 135.28 micromoles, 1 equivalent), sodium tert-butoxide (39 mg, 405.83 micromoles, 3 equivalents), 2- Dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (12.9 mg, 27.06 micromoles, 0.2 equiv), tris(dibenzylideneacetone)dipalladium (12.39 mg, 13.53 micromoles, 0.1 equiv) Dissolve in dioxane (2 milliliters), then react at 120 degrees centigrade under microwave for 1.5 hours, after the reaction is completed, add water (5 milliliters) to the reaction solution and extract with ethyl acetate (5 milliliters × 2), the organic phase is used Washed with saturated brine (10 ml), dried over anhydrous sodium sulfate, filtered, and the concentrated residue was subjected to silica gel thin-layer chromatography (dichloromethane:methanol=10:1) and high performance liquid chromatography (chromatographic column: Phenomenex Synergi C18 150*25 mm*10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 7%-37%, 10 minutes) separation and purification to obtain compound 15.
MS-ESI计算值[M+H] +486，实测值486。 MS-ESI calculated [M+H] + 486, found 486.
1H NMR(400MHz,CD 3OD)δ＝8.36(dd,J＝8.44,4.52Hz,1H)8.09-8.01(m,1H)7.53(ddd,J＝14.31,7.70,1.22Hz,1H)7.48-7.40(m,1H)7.2.-7.15(m,1H)6.42-6.33(m,1H)4.24(s,4H)3.93-3.80(m,3H)1.90-1.78(m,6H)0.83-0.73(m,4H). 1 H NMR (400MHz, CD 3 OD) δ = 8.36 (dd, J = 8.44, 4.52Hz, 1H) 8.09-8.01 (m, 1H) 7.53 (ddd, J = 14.31, 7.70, 1.22Hz, 1H) 7.48- 7.40(m,1H)7.2.-7.15(m,1H)6.42-6.33(m,1H)4.24(s,4H)3.93-3.80(m,3H)1.90-1.78(m,6H)0.83-0.73(m ,4H).
实施例16Example 16
Figure PCTCN2022129958-appb-000063
Figure PCTCN2022129958-appb-000063
第一步first step
将化合物15-4(400毫克，1.68毫摩尔，1当量)，F(400.91毫克2.58毫摩尔，1.53当量)，N,N-二异丙基乙胺(869.99毫克，6.73毫摩尔，4当量)溶于乙二醇(10毫升)，接着在140摄氏度微波下反应1.5小时。反应完毕后，向反应液中加入水(50毫升)乙酸乙酯萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经硅胶薄层色谱板(石油醚：乙酸乙酯＝1:1)分离纯化得到化合物16-1。Compound 15-4 (400 mg, 1.68 mmol, 1 eq), F (400.91 mg, 2.58 mmol, 1.53 eq), N,N-diisopropylethylamine (869.99 mg, 6.73 mmol, 4 eq) Dissolved in ethylene glycol (10 mL), followed by reaction under microwave at 140°C for 1.5 hours. After the reaction was completed, water (50 ml) was added to the reaction solution to extract with ethyl acetate (50 ml×2), the organic phase was washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the concentrated residue was washed with Compound 16-1 was obtained by separation and purification on a silica gel thin-layer chromatography plate (petroleum ether: ethyl acetate = 1:1).
MS-ESI计算值[M+H] +321，实测值321。 MS-ESI calculated [M+H] +321 , found 321.
1H NMR(400MHz,CDCl 3)δ＝6.02(s,1H)5.94-5.84(m,2H)4.48-4.40(m,2H)4.32-4.25(m,2H)3.86-3.81(m,3H)2.05–1.92(m,6H)1.64-1.58(m,2H). 1 H NMR (400MHz, CDCl 3 )δ=6.02(s,1H)5.94-5.84(m,2H)4.48-4.40(m,2H)4.32-4.25(m,2H)3.86-3.81(m,3H)2.05 –1.92(m,6H)1.64-1.58(m,2H).
第二步second step
将化合物16-1(237毫克，739.85微摩尔，1当量)，盐酸羟胺(514.13毫克，7.40毫摩尔，10当量)，氢氧化钾(93.03毫克，1.48毫摩尔，2当量)溶于乙醇(7毫升)水(3.5毫升)中，接着在120摄氏度微波下反应1.5小时反应完毕后，向反应液中加入水(20毫升)二氯甲烷萃取(20毫升×3)，有机相用饱和的食盐水(20毫升×2)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝10:1)分离纯化得到化合物16-2。Compound 16-1 (237 mg, 739.85 micromole, 1 eq), hydroxylamine hydrochloride (514.13 mg, 7.40 mmol, 10 eq), potassium hydroxide (93.03 mg, 1.48 mmol, 2 eq) were dissolved in ethanol (7 milliliters) in water (3.5 milliliters), then reacted at 120 degrees centigrade for 1.5 hours under microwaves. (20 ml×2), dried over anhydrous sodium sulfate, filtered, and the concentrated residue was separated and purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1) to obtain compound 16-2.
MS-ESI计算值[M+H] +243，实测值243。 MS-ESI calculated [M+H] +243 , found 243.
1H NMR(400MHz,CD 3OD)δ＝6.29-6.27(m,1H)4.21-4.14(m,2H)4.12-4.06(m,2H)3.97-3.90(m,3H)1.63-1.54(m,2H)。 1 H NMR (400MHz, CD 3 OD) δ=6.29-6.27(m,1H)4.21-4.14(m,2H)4.12-4.06(m,2H)3.97-3.90(m,3H)1.63-1.54(m, 2H).
第三步third step
将化合物16-2(70毫克，268.76微摩尔，1当量)，中间体E(127.44克，403.14微摩尔，1.5当量)，叔丁醇钠(77.48毫克，806.28微摩尔，3当量)，2-二环己基磷-2,4,6-三异丙基联苯(25.62毫克，53.75微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(24.61毫克，26.88微摩尔，0.1当量)溶于二氧六环(7毫升)中，接着在氮气保护下110摄氏度搅拌12小时，反应完毕后，向反应液中加入水(25毫升)乙酸乙酯萃取(25毫升×2)，有机相用饱和的食盐水(25毫升)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝10:1)和高效液相色谱法(色谱柱：Phenomenex Synergi  C18 150*25毫米*10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：8％-38％，10分钟)分离纯化得到化合物16。Compound 16-2 (70 mg, 268.76 micromoles, 1 equivalent), Intermediate E (127.44 grams, 403.14 micromoles, 1.5 equivalents), sodium tert-butoxide (77.48 mg, 806.28 micromoles, 3 equivalents), 2- Dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (25.62 mg, 53.75 micromoles, 0.2 equiv), tris(dibenzylideneacetone)dipalladium (24.61 mg, 26.88 micromoles, 0.1 equiv) Dissolve in dioxane (7 milliliters), then stir at 110 degrees centigrade under nitrogen protection for 12 hours, after the reaction is complete, add water (25 milliliters) to the reaction solution and extract with ethyl acetate (25 milliliters × 2), the organic phase Washed with saturated brine (25 ml), dried over anhydrous sodium sulfate, filtered, and the concentrated residue was subjected to silica gel thin-layer chromatography (dichloromethane:methanol=10:1) and high performance liquid chromatography (chromatographic column: Phenomenex Synergi C18 150*25 mm*10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 8%-38%, 10 minutes) separation and purification to obtain compound 16.
MS-ESI计算值[M+H] +522，实测值522。 MS-ESI calculated [M+H] + 522, found 522.
1H NMR(400MHz,CD 3OD)δ＝8.47-8.39(m,1H)8.20-8.11(m,1H)7.62-7.48(m,2H)7.28-7.20(m,1H)6.62-6.55(m,1H)4.42-4.35(m,2H)4.32-4.25(m,2H)4.00–3.94(m,3H)1.88-1.86(m,3H)1.85-1.83(m,3H)1.73-1.67(m,2H)。 1 H NMR (400MHz, CD 3 OD) δ=8.47-8.39(m,1H)8.20-8.11(m,1H)7.62-7.48(m,2H)7.28-7.20(m,1H)6.62-6.55(m, 1H)4.42-4.35(m,2H)4.32-4.25(m,2H)4.00–3.94(m,3H)1.88-1.86(m,3H)1.85-1.83(m,3H)1.73-1.67(m,2H) .
实施例17Example 17
Figure PCTCN2022129958-appb-000064
Figure PCTCN2022129958-appb-000064
第一步first step
将化合物17-1(2.00克，11.82毫摩尔，1当量)溶于1,2-二氯乙烷(10毫升)，接着向反应液中加入过氧化苯甲酰(286.40毫克，1.18毫摩尔，0.1当量)，N-溴代丁二酰亚胺(4.63克，26.01毫摩尔，2.2当量)，在90摄氏度下反应2小时。反应完成后，加饱和硫代硫酸钠水溶液(50毫升)，乙酸乙酯萃取(20毫升×3)，合并的有机相用饱和食盐水溶液(45毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝1：0)纯化得到化合物17-2。Compound 17-1 (2.00 g, 11.82 mmol, 1 eq) was dissolved in 1,2-dichloroethane (10 ml), and then benzoyl peroxide (286.40 mg, 1.18 mmol, 0.1 equivalent), N-bromosuccinimide (4.63 grams, 26.01 mmol, 2.2 equivalents), reacted at 90 degrees Celsius for 2 hours. After the reaction was complete, add saturated aqueous sodium thiosulfate solution (50 ml), extract with ethyl acetate (20 ml × 3), wash the combined organic phase with saturated saline solution (45 ml × 2), and wash the organic phase with anhydrous sodium sulfate After drying, filtering, and concentrating under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:0) to obtain compound 17-2.
1HNMR(400MHz,CDCl 3)δ＝7.97-7.90(m,1H)7.16(t,J＝8.68Hz,1H)4.80(s,2H)4.62(d,J＝1.83Hz,2H). 1 HNMR (400MHz, CDCl 3 )δ=7.97-7.90(m,1H)7.16(t,J=8.68Hz,1H)4.80(s,2H)4.62(d,J=1.83Hz,2H).
第二步second step
将化合物17-2(5.75克，17.59毫摩尔，1当量)溶于甲苯(60毫升)，接着向反应液中加入三氧化二铝(72.09克，707.00毫摩尔，40.2当量)，水(633.67毫克，35.17毫摩尔，633.67微升，2当量)，在120摄氏度下反应2小时。反应完成后，反应液过滤，滤液浓缩，加水(200毫升)稀释，乙酸乙酯萃取(100毫升×3)，有机相用无水硫酸钠干燥，过滤，浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝1:0-200:1)纯化得到化合物17-3。Compound 17-2 (5.75 g, 17.59 mmol, 1 eq) was dissolved in toluene (60 ml), then aluminum oxide (72.09 g, 707.00 mmol, 40.2 eq), water (633.67 mg , 35.17 mmol, 633.67 microliters, 2 equivalents), reacted at 120 degrees Celsius for 2 hours. After the reaction was completed, the reaction liquid was filtered, the filtrate was concentrated, diluted with water (200 ml), extracted with ethyl acetate (100 ml x 3), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:0-200:1) to obtain compound 17-3.
1HNMR(400MHz,CDCl 3)δ＝8.20(dd,J＝8.93,4.16Hz,1H),7.19-7.10(m,1H),5.55(t,J＝1.90Hz,2H),5.25(s,2H). 1 HNMR (400MHz, CDCl 3 ) δ＝8.20(dd, J＝8.93, 4.16Hz, 1H), 7.19-7.10(m, 1H), 5.55(t, J＝1.90Hz, 2H), 5.25(s, 2H ).
第三步third step
将化合物17-3(300.00毫克，1.64毫摩尔，1当量)溶于N,N-二甲基甲酰胺(5毫升)，接着向反应液 中加入碳酸钾(452.80毫克，3.28毫摩尔，2当量)，I(238.73毫克，2.46毫摩尔，1.5当量)，在80摄氏度下反应2小时。反应完成后，加水(30毫升)稀释，乙酸乙酯萃取(15毫升×3)，有机相用饱和食盐水(25毫升×3)洗涤，无水硫酸钠干燥，过滤，浓缩得到化合物17-4。Compound 17-3 (300.00 mg, 1.64 mmol, 1 eq) was dissolved in N,N-dimethylformamide (5 ml), and potassium carbonate (452.80 mg, 3.28 mmol, 2 eq. ), I (238.73 mg, 2.46 mmol, 1.5 equivalents), reacted at 80 degrees Celsius for 2 hours. After the reaction was completed, it was diluted with water (30 ml), extracted with ethyl acetate (15 ml × 3), the organic phase was washed with saturated brine (25 ml × 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 17-4 .
MS-ESI计算值[M+H] +261,实测值261。 MS-ESI calculated value [M+H] + 261, found value 261.
第四步the fourth step
将化合物17-4(420.00毫克，1.48毫摩尔，1当量)溶于甲醇(10毫升)，接着向反应液中加入钯碳(100.00毫克，纯度：10％)，在25摄氏度氢气氛围(15PSI)下反应2小时。反应完成后，垫硅藻土过滤，滤液浓缩得到化合物17-5。Compound 17-4 (420.00 mg, 1.48 mmol, 1 eq) was dissolved in methanol (10 ml), then palladium carbon (100.00 mg, purity: 10%) was added to the reaction solution, and the mixture was dissolved in a hydrogen atmosphere (15PSI) at 25 degrees Celsius. The reaction was carried out for 2 hours. After the reaction was completed, it was filtered with celite, and the filtrate was concentrated to obtain compound 17-5.
MS-ESI计算值[M+H] +230,实测值230。 MS-ESI calculated value [M+H] + 230, found value 230.
第五步the fifth step
将化合物17-5(200.35毫克，869.92微摩尔，1.1当量)，中间体E(250.00毫克，790.83微摩尔，1当量)溶于四氢呋喃(6毫升)，接着向反应液中加入叔丁醇钠(152.00毫克，1.58毫摩尔，2当量)，三(二亚苄基丙酮)二钯(72.42毫克，79.08微摩尔，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(75.40毫克，158.17微摩尔，0.2当量)，在70摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶薄层色谱板(石油醚：乙酸乙酯：乙醇＝4：3：1)纯化得到的粗品经高效液相色谱(色谱柱：Waters Xbridge C18 150*50毫米*10微米；流动相：流动相A：碳酸氢铵水溶液；流动相B：乙腈；B％:37％-67％，11分钟)分离得到化合物17。Compound 17-5 (200.35 mg, 869.92 micromoles, 1.1 equivalents), Intermediate E (250.00 mg, 790.83 micromoles, 1 equivalents) were dissolved in tetrahydrofuran (6 ml), and sodium tert-butoxide ( 152.00 mg, 1.58 mmol, 2 equiv), tris(dibenzylideneacetone) dipalladium (72.42 mg, 79.08 micromol, 0.1 equiv), 2-dicyclohexylphosphonium-2,4,6-triisopropyl Biphenyl (75.40 mg, 158.17 micromole, 0.2 eq.) was reacted at 70°C for 12 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by silica gel thin-layer chromatography (petroleum ether: ethyl acetate: ethanol = 4:3:1) and the crude product was purified by high performance liquid chromatography (column: Waters Xbridge C18 150*50 mm*10 microns; mobile phase: Mobile phase A: ammonium bicarbonate aqueous solution; mobile phase B: acetonitrile; B%: 37%-67%, 11 minutes) to obtain compound 17.
MS-ESI计算值[M+H] +510,实测值510。 MS-ESI calculated value [M+H] + 510, found value 510.
1HNMR(400MHz,CD 3OD)δ＝8.40-8.29(m,1H),7.99(s,1H),7.59-7.51(m,1H),7.34(br t,J＝7.83Hz,1H),7.23-7.16(m,1H),7.13(d,J＝8.31Hz,1H),6.40(d,J＝8.31Hz,1H),5.06(s,2H),4.62(s,1H),3.85(s,4H),2.26(t,J＝7.64Hz,4H),1.99-1.90(m,2H),1.87(s,3H),1.84(s,3H). 1 HNMR (400MHz, CD 3 OD) δ = 8.40-8.29 (m, 1H), 7.99 (s, 1H), 7.59-7.51 (m, 1H), 7.34 (br t, J = 7.83Hz, 1H), 7.23 -7.16(m,1H),7.13(d,J=8.31Hz,1H),6.40(d,J=8.31Hz,1H),5.06(s,2H),4.62(s,1H),3.85(s, 4H), 2.26(t, J=7.64Hz, 4H), 1.99-1.90(m, 2H), 1.87(s, 3H), 1.84(s, 3H).
实施例18Example 18
Figure PCTCN2022129958-appb-000065
Figure PCTCN2022129958-appb-000065
第一步first step
将化合物17-3(300毫克，1.64毫摩尔，1当量)溶于乙腈(10毫升)，接着向反应液中加入中间体A (402.11毫克，2.46毫摩尔，1.5当量)，碳酸钾(679.20毫克,4.91毫摩尔,3eq)，在80摄氏度下反应2小时。反应完成后，加入乙酸乙酯(30毫升×3)有机相经饱和食盐水(20毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩得到化合物18-1。Compound 17-3 (300 mg, 1.64 mmol, 1 eq) was dissolved in acetonitrile (10 ml), then Intermediate A (402.11 mg, 2.46 mmol, 1.5 eq), potassium carbonate (679.20 mg , 4.91 mmol, 3eq), reacted at 80 degrees Celsius for 2 hours. After the reaction was complete, ethyl acetate (30 mL×3) was added to the organic phase, washed with saturated brine (20 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 18-1.
MS-ESI计算值[M+H] +291,实测值291。 MS-ESI calculated value [M+H] + 291, found value 291.
第三步third step
将化合物18-1(400毫克，1.38毫摩尔，1当量)溶于甲醇(10毫升)，接着向反应液中加入钯碳(50毫克，纯度：10％)，在25摄氏度氢气氛围(15PSI)下反应1小时。反应完成后，垫硅藻土过滤，滤液浓缩得到化合物18-2。Compound 18-1 (400 mg, 1.38 mmol, 1 eq) was dissolved in methanol (10 ml), then palladium on carbon (50 mg, purity: 10%) was added to the reaction solution, and the compound was dissolved in a hydrogen atmosphere (15PSI) at 25 degrees Celsius. The reaction was carried out for 1 hour. After the reaction was completed, it was filtered with celite, and the filtrate was concentrated to obtain compound 18-2.
MS-ESI计算值[M+H] +261,实测值261。 MS-ESI calculated value [M+H] + 261, found value 261.
第四步the fourth step
将化合物18-2(300毫克，1.15毫摩尔，1.1当量)溶于四氢呋喃(10毫升)，接着加入中间体E(331.18毫克，1.05毫摩尔，1当量)并在氮气保护下向反应液中加入三(二亚苄基丙酮)二钯(95.93毫克，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(99.88毫克,0.2当量)，叔丁醇钠(201.35毫克，2当量)，在70摄氏度下反应12小时。反应完成后，乙酸乙酯萃取(30毫升×3)，有机相用饱和食盐水(20毫升×3)洗涤，无水硫酸钠干燥，过滤，浓缩。粗品经高效液相色谱(色谱柱：Waters Xbridge C18 150*50毫米*10微米；流动相A：碳酸氢铵水溶液，流动相B：乙腈；B％:28％-58％，11分钟)分离得到化合物18。Compound 18-2 (300 mg, 1.15 mmol, 1.1 eq) was dissolved in tetrahydrofuran (10 ml), then intermediate E (331.18 mg, 1.05 mmol, 1 eq) was added and added to the reaction solution under nitrogen protection Tris(dibenzylideneacetone)dipalladium (95.93 mg, 0.1 equivalent), 2-dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (99.88 mg, 0.2 equivalent), sodium tert-butoxide ( 201.35 mg, 2 equivalents), reacted at 70 degrees Celsius for 12 hours. After the reaction was complete, it was extracted with ethyl acetate (30 mL×3), and the organic phase was washed with saturated brine (20 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated by high performance liquid chromatography (column: Waters Xbridge C18 150*50 mm*10 microns; mobile phase A: ammonium bicarbonate aqueous solution, mobile phase B: acetonitrile; B%: 28%-58%, 11 minutes) to obtain Compound 18.
MS-ESI计算值[M+H] +540,实测值540。 MS-ESI calculated value [M+H] + 540, found value 540.
1H-NMR(400MHz,CD 3OD)δ＝8.33(br s,1H),7.99(s,1H),7.55(ddd,J＝1.4,7.7,14.3Hz,1H),7.34(br t,J＝7.8Hz,1H),7.20(t,J＝7.2Hz,1H),7.14(d,J＝8.4Hz,1H),6.39(d,J＝8.4Hz,1H),5.08-4.93(m,3H),4.61(s,3H),4.68-4.52(m,1H),3.93-3.81(m,5H),3.31-3.24(m,4H),2.61-2.53(m,2H),2.19-2.11(m,2H),1.86(d,J＝13.4Hz,3H),1.88(br d,J＝13.5Hz,3H). 1 H-NMR (400MHz, CD 3 OD) δ = 8.33 (br s, 1H), 7.99 (s, 1H), 7.55 (ddd, J = 1.4, 7.7, 14.3Hz, 1H), 7.34 (br t, J =7.8Hz, 1H), 7.20(t, J=7.2Hz, 1H), 7.14(d, J=8.4Hz, 1H), 6.39(d, J=8.4Hz, 1H), 5.08-4.93(m, 3H ),4.61(s,3H),4.68-4.52(m,1H),3.93-3.81(m,5H),3.31-3.24(m,4H),2.61-2.53(m,2H),2.19-2.11(m ,2H),1.86(d,J=13.4Hz,3H),1.88(br d,J=13.5Hz,3H).
实施例19Example 19
Figure PCTCN2022129958-appb-000066
Figure PCTCN2022129958-appb-000066
第一步first step
将化合物17-3(300毫克，1.64毫摩尔，1当量)和中间体D(391.81毫克，3.28毫摩尔，2当量)溶于乙腈(10毫升)，接着向反应液中加入碳酸钾(679.2毫克，4.9毫摩尔，3当量)，在80摄氏度氮气保护下反应2小时。反应完成后，乙酸乙酯萃取(10毫升×3)，饱和食盐水(5毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩得到化合物19-1。Compound 17-3 (300 mg, 1.64 mmol, 1 eq) and intermediate D (391.81 mg, 3.28 mmol, 2 eq) were dissolved in acetonitrile (10 ml), then potassium carbonate (679.2 mg , 4.9 mmol, 3 equivalents), reacted for 2 hours at 80 degrees Celsius under nitrogen protection. After the reaction was completed, the mixture was extracted with ethyl acetate (10 mL×3), washed with saturated brine (5 mL×2), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 19-1.
MS-ESI计算值[M+H] +247,实测值247。 MS-ESI calculated [M+H] + 247, found 247.
第二步second step
将化合物19-1(400.00毫克，1.62毫摩尔，1当量)溶于甲醇(5毫升)，接着向反应液中加入钯碳(50.00毫克，纯度：10％)，在25摄氏度氢气氛围(15PSI)下反应2小时。反应完成后，垫硅藻土过滤，滤液浓缩得到化合物19-2。Compound 19-1 (400.00 mg, 1.62 mmol, 1 equivalent) was dissolved in methanol (5 ml), then palladium on carbon (50.00 mg, purity: 10%) was added to the reaction solution, and the compound was dissolved in a hydrogen atmosphere (15PSI) at 25 degrees Celsius. The reaction was carried out for 2 hours. After the reaction was completed, it was filtered with celite, and the filtrate was concentrated to obtain compound 19-2.
MS-ESI计算值[M+H] +217,实测值217。 MS-ESI calculated [M+H] + 217, found 217.
第三步third step
将化合物19-2(200毫克，924.7微摩尔，1.1当量)溶于四氢呋喃(10毫升)，接着向反应液中加入中间体E(265.75毫克，840.67微摩尔，1当量)，叔丁醇钠(161.58毫克，1.68毫摩尔，2当量)，在20摄氏度氮气保护下加入三(二亚苄基丙酮)二钯(76.98毫克，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(80.15毫克，0.2当量)，然后加热至70摄氏度反应12小时。反应完成后，过滤，加入水(10毫升)，乙酸乙酯(30毫升×2)，有机相经饱和食盐水(10毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩。粗品经高效液相色谱(色谱柱：Waters Xbridge 150*25毫米*5微米；流动相A:氨水，流动相B：乙腈；B％：30％-60％，9分钟)分离得到化合物19。Compound 19-2 (200 mg, 924.7 micromoles, 1.1 eq) was dissolved in tetrahydrofuran (10 ml), then intermediate E (265.75 mg, 840.67 micromoles, 1 eq), sodium tert-butoxide ( 161.58 mg, 1.68 mmol, 2 equivalents), added tris(dibenzylideneacetone) dipalladium (76.98 mg, 0.1 equivalents), 2-dicyclohexylphosphine-2,4,6-tris Isopropyl biphenyl (80.15 mg, 0.2 equivalents), then heated to 70 degrees Celsius for 12 hours. After completion of the reaction, filter, add water (10 ml), ethyl acetate (30 ml × 2), the organic phase was washed with saturated brine (10 ml × 2), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated by high performance liquid chromatography (column: Waters Xbridge 150*25 mm*5 microns; mobile phase A: ammonia water, mobile phase B: acetonitrile; B%: 30%-60%, 9 minutes) to obtain compound 19.
MS-ESI计算值[M+H] +496,实测值496。 MS-ESI calculated [M+H] + 496, found 496.
1H NMR(400MHz,CDCl 3)δ＝10.97(s,1H),8.56(dd,J＝4.5,8.5Hz,1H),8.05(s,1H),7.40-7.34(m,1H),7.33-7.29(m,1H),7.27-7.22(m,1H),7.12-7.04(m,1H),6.44(s,1H),6.36(d,J＝8.4Hz,1H),5.14(s,2H),5.04(s,2H),4.02(s,4H),1.86(s,3H),1.83(s,3H),0.71(s,4H). 1 H NMR (400MHz, CDCl 3 )δ=10.97(s,1H),8.56(dd,J=4.5,8.5Hz,1H),8.05(s,1H),7.40-7.34(m,1H),7.33- 7.29(m,1H),7.27-7.22(m,1H),7.12-7.04(m,1H),6.44(s,1H),6.36(d,J＝8.4Hz,1H),5.14(s,2H) ,5.04(s,2H),4.02(s,4H),1.86(s,3H),1.83(s,3H),0.71(s,4H).
实施例20Example 20
Figure PCTCN2022129958-appb-000067
Figure PCTCN2022129958-appb-000067
第一步first step
将化合物17-3(350.00毫克，1.91毫摩尔，1当量)溶于N,N-二甲基甲酰胺(5毫升)，接着向反应液中加入碳酸钾(528.26毫克，3.82毫摩尔，2当量)，G(291.68毫克，2.29毫摩尔，1.2当量)，在80摄氏度下反应2小时。反应完成后，加水(30毫升)稀释，乙酸乙酯萃取(15毫升×3)，有机相用饱和食盐水(25毫升×3)洗涤，无水硫酸钠干燥，过滤，浓缩得到化合物20-1。Compound 17-3 (350.00 mg, 1.91 mmol, 1 eq) was dissolved in N,N-dimethylformamide (5 ml), and then potassium carbonate (528.26 mg, 3.82 mmol, 2 eq ), G (291.68 mg, 2.29 mmol, 1.2 equivalents), reacted at 80 degrees Celsius for 2 hours. After the reaction was completed, it was diluted with water (30 ml), extracted with ethyl acetate (15 ml × 3), the organic phase was washed with saturated brine (25 ml × 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 20-1 .
MS-ESI计算值[M+H] +291,实测值291。 MS-ESI calculated value [M+H] + 291, found value 291.
第二步second step
将化合物20-1(500.00毫克，1.72毫摩尔，1当量)溶于甲醇(10毫升)，接着向反应液中加入钯碳(50.00毫克，纯度：10％)，在25摄氏度氢气氛围(15PSI)下反应2小时。反应完成后，垫硅藻土过滤，滤液浓缩得到化合物20-2。Compound 20-1 (500.00 mg, 1.72 mmol, 1 eq) was dissolved in methanol (10 mL), then palladium carbon (50.00 mg, purity: 10%) was added to the reaction solution, and the mixture was dissolved in a hydrogen atmosphere (15PSI) at 25 degrees Celsius. The reaction was carried out for 2 hours. After the reaction was completed, it was filtered with celite, and the filtrate was concentrated to obtain compound 20-2.
MS-ESI计算值[M+H] +261,实测值261。 MS-ESI calculated value [M+H] + 261, found value 261.
第三步third step
将化合物20-2(398.58毫克，1.53毫摩尔，1.1当量)，中间体E(440.00毫克，1.39毫摩尔，1当量)溶于四氢呋喃(10毫升)，接着向反应液中加入叔丁醇钠(267.52毫克，2.78毫摩尔，2当量)，三(二亚苄基丙酮)二钯(127.46毫克，139.19微摩尔，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(132.71毫克，278.37微摩尔，0.2当量)，在70摄氏度下反应12小时。反应完成后，反应液过滤，滤液用乙酸乙酯萃取(20毫升×3)，有机相用饱和食盐水(50毫升×3)洗涤，无水硫酸钠干燥，过滤，浓缩的粗品经硅胶柱层析(石油醚：乙酸乙酯：乙醇＝40：3：1-4：3：1)纯化得到的粗品经高效液相色谱(色谱柱：Waters Xbridge C18 150*50毫米*10微米；流动相：流动相A：碳酸氢铵水溶液；流动相B：乙腈；B％:26％-56％，11分钟)分离得到化合物20。Compound 20-2 (398.58 mg, 1.53 mmol, 1.1 eq), Intermediate E (440.00 mg, 1.39 mmol, 1 eq) were dissolved in tetrahydrofuran (10 ml), and sodium tert-butoxide ( 267.52 mg, 2.78 mmol, 2 equiv), tris(dibenzylideneacetone) dipalladium (127.46 mg, 139.19 micromol, 0.1 equiv), 2-dicyclohexylphosphine-2,4,6-triisopropyl Biphenyl (132.71 mg, 278.37 micromole, 0.2 eq.) was reacted at 70°C for 12 hours. After the reaction was completed, the reaction liquid was filtered, and the filtrate was extracted with ethyl acetate (20 ml × 3), the organic phase was washed with saturated brine (50 ml × 3), dried over anhydrous sodium sulfate, filtered, and the concentrated crude product was passed through a silica gel column layer. Analysis (petroleum ether: ethyl acetate: ethanol = 40: 3: 1-4: 3: 1) the crude product purified by high performance liquid chromatography (column: Waters Xbridge C18 150 * 50 mm * 10 microns; mobile phase: Mobile phase A: ammonium bicarbonate aqueous solution; mobile phase B: acetonitrile; B%: 26%-56%, 11 minutes) to obtain compound 20.
MS-ESI计算值[M+H] +540,实测值540。 MS-ESI calculated value [M+H] + 540, found value 540.
1H-NMR(400MHz,CD 3OD)δ＝8.38-8.26(m,1H),8.02-7.97(m,1H),7.60-7.49(m,1H),7.40-7.31(m,1H),7.23-7.16(m,1H),7.16-7.10(m,1H),6.45-6.38(m,1H),5.12-5.07(m,2H),4.91(br s,2H),4.59-4.58(m,1H),3.72(s,4H),3.71-3.68(m,2H),2.08-2.04(m,2H),1.89-1.86(m,6H),1.85-1.84(m,2H). 1 H-NMR (400MHz, CD 3 OD) δ=8.38-8.26(m,1H),8.02-7.97(m,1H),7.60-7.49(m,1H),7.40-7.31(m,1H),7.23 -7.16(m,1H),7.16-7.10(m,1H),6.45-6.38(m,1H),5.12-5.07(m,2H),4.91(br s,2H),4.59-4.58(m,1H ),3.72(s,4H),3.71-3.68(m,2H),2.08-2.04(m,2H),1.89-1.86(m,6H),1.85-1.84(m,2H).
实施例21Example 21
Figure PCTCN2022129958-appb-000068
Figure PCTCN2022129958-appb-000068
第一步first step
将化合物17-3(362.38毫克，1.98毫摩尔，1当量)溶于N,N-二甲基甲酰胺(12毫升)，接着向反应液中加入碳酸钾(820.46毫克，5.94毫摩尔，3当量)，B(300.00毫克，1.98毫摩尔，1当量)，在80摄氏度下反应2小时。反应完成后，加水(20毫升)稀释，过滤，滤液浓缩得到化合物21-1。Compound 17-3 (362.38 mg, 1.98 mmol, 1 eq.) was dissolved in N,N-dimethylformamide (12 ml), and then potassium carbonate (820.46 mg, 5.94 mmol, 3 eq. ), B (300.00 mg, 1.98 mmol, 1 equivalent), reacted at 80 degrees Celsius for 2 hours. After the reaction was completed, it was diluted with water (20 mL), filtered, and the filtrate was concentrated to obtain compound 21-1.
MS-ESI计算值[M+H] +279,实测值279。 MS-ESI calculated [M+H] + 279, found 279.
第二步second step
将化合物21-1(300.00毫克，1.08毫摩尔，1当量)溶于甲醇(10毫升)，接着向反应液中加入钯碳(50.00毫克，纯度：10％)，在25摄氏度氢气氛围(50PSI)下反应2小时。反应完成后，垫硅藻土过滤，滤液浓缩得到化合物21-2。Compound 21-1 (300.00 mg, 1.08 mmol, 1 eq) was dissolved in methanol (10 ml), then palladium carbon (50.00 mg, purity: 10%) was added to the reaction solution, and the The reaction was carried out for 2 hours. After the reaction was completed, it was filtered with celite, and the filtrate was concentrated to obtain compound 21-2.
MS-ESI计算值[M+H] +249,实测值249。 MS-ESI calculated [M+H] + 249, found 249.
第三步third step
将化合物21-2(150.00毫克，604.12微摩尔，1当量)，中间体E(190.98毫克，604.12微摩尔，1当量)溶于四氢呋喃(10毫升)，接着向反应液中加入叔丁醇钠(116.11毫克，1.21毫摩尔，2当量)，三(二亚苄基丙酮)二钯(55.32毫克，60.41微摩尔，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(57.60毫克，120.82微摩尔，0.2当量)，在70摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶薄层色谱板(石油醚：乙酸乙酯＝0：1)纯化得到的粗品进一步经高效液相色谱(色谱柱：Waters Xbridge 150*25毫米*5微米；流动相：流动相A：碳酸氢铵水溶液；流动相B：乙腈；B％:38％-68％，10分钟)分离得到化合物21。Compound 21-2 (150.00 mg, 604.12 micromoles, 1 equivalent), Intermediate E (190.98 mg, 604.12 micromoles, 1 equivalent) were dissolved in tetrahydrofuran (10 ml), and then sodium tert-butoxide was added to the reaction solution ( 116.11 mg, 1.21 mmol, 2 equiv), tris(dibenzylideneacetone)dipalladium (55.32 mg, 60.41 micromol, 0.1 equiv), 2-dicyclohexylphosphine-2,4,6-triisopropyl Biphenyl (57.60 mg, 120.82 micromole, 0.2 equiv), reacted at 70 degrees Celsius for 12 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by silica gel thin-layer chromatography plate (petroleum ether: ethyl acetate=0:1), and the crude product was further subjected to high performance liquid chromatography (column: Waters Xbridge 150*25 mm*5 microns; mobile phase: mobile phase A: Aqueous solution of ammonium bicarbonate; mobile phase B: acetonitrile; B%: 38%-68%, 10 minutes) isolated to obtain compound 21.
MS-ESI计算值[M+H] +528,实测值528。 MS-ESI calculated value [M+H] + 528, found value 528.
1HNMR(400MHz,CDCl 3)δ＝11.13-11.09(m,1H),11.06-10.95(m,1H),8.59-8.48(m,1H),8.09-7.97(m,1H),7.38-7.30(m,2H),7.26-7.20(m,1H),7.14-6.99(m,1H),6.54-6.38(m,1H),6.33-6.27(m,1H),5.11-4.95(m,5H),3.93-3.90(m,2H),3.86(s,2H),2.72-2.61(m,2H),2.53-2.39(m,2H),1.85(s,3H),1.82(s,3H). 1 HNMR (400MHz, CDCl 3 ) δ=11.13-11.09 (m, 1H), 11.06-10.95 (m, 1H), 8.59-8.48 (m, 1H), 8.09-7.97 (m, 1H), 7.38-7.30 ( m,2H),7.26-7.20(m,1H),7.14-6.99(m,1H),6.54-6.38(m,1H),6.33-6.27(m,1H),5.11-4.95(m,5H), 3.93-3.90(m,2H),3.86(s,2H),2.72-2.61(m,2H),2.53-2.39(m,2H),1.85(s,3H),1.82(s,3H).
实施例22Example 22
Figure PCTCN2022129958-appb-000069
Figure PCTCN2022129958-appb-000069
第一步first step
将中间体C(220.00毫克，1.30毫摩尔，1当量)溶于N,N-二甲基甲酰胺(1毫升)，接着向反应液中加入碳酸钾(537.85毫克，3.89毫摩尔，3当量)，17-3(237.56毫克，1.30毫摩尔，1当量)，在80摄氏度下反应2小时。反应完成后，加水(20毫升)稀释，过滤，滤饼干燥得到化合物22-1。Intermediate C (220.00 mg, 1.30 mmol, 1 eq) was dissolved in N,N-dimethylformamide (1 mL), then potassium carbonate (537.85 mg, 3.89 mmol, 3 eq) was added to the reaction , 17-3 (237.56 mg, 1.30 mmol, 1 equiv), reacted at 80°C for 2 hours. After the reaction was completed, it was diluted with water (20 ml), filtered, and the filter cake was dried to obtain compound 22-1.
MS-ESI计算值[M+H] +297,实测值297。 MS-ESI calculated [M+H] +297 , found 297.
第二步second step
将化合物22-1(300.00毫克，1.01毫摩尔，1当量)溶于甲醇(10毫升)，接着向反应液中加入钯碳(50.00毫克，纯度：10％)，在25摄氏度氢气氛围(15PSI)下反应2小时。反应完成后，垫硅藻土过滤，滤液浓缩得到化合物22-2。Compound 22-1 (300.00 mg, 1.01 mmol, 1 eq) was dissolved in methanol (10 ml), then palladium on carbon (50.00 mg, purity: 10%) was added to the reaction solution, and the compound was dissolved in a hydrogen atmosphere (15PSI) at 25 degrees Celsius. The reaction was carried out for 2 hours. After the reaction was completed, it was filtered with celite, and the filtrate was concentrated to obtain compound 22-2.
MS-ESI计算值[M+H] +267,实测值267。 MS-ESI calculated [M+H] +267 , found 267.
第三步third step
将化合物22-2(207.22毫克，778.18微摩尔，1.2当量)，中间体E(205.00毫克，648.48微摩尔，1当量)溶于四氢呋喃(10毫升)，接着向反应液中加入叔丁醇钠(124.64毫克，1.30毫摩尔，2当量)，三(二亚苄基丙酮)二钯(59.38毫克，64.85微摩尔，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(61.83毫克，129.70微摩尔，0.2当量)，在70摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)纯化得到的粗品进一步经高效液相色谱(色谱柱：Waters Xbridge C18 150*50毫米*10微米；流动相：流动相A：碳酸氢铵水溶液；流动相B：乙腈；B％:33％-63％，11分钟)分离得到化合物22。Compound 22-2 (207.22 mg, 778.18 micromoles, 1.2 equivalents), Intermediate E (205.00 mg, 648.48 micromoles, 1 equivalents) were dissolved in tetrahydrofuran (10 ml), and sodium tert-butoxide ( 124.64 mg, 1.30 mmol, 2 equiv), tris(dibenzylideneacetone) dipalladium (59.38 mg, 64.85 micromol, 0.1 equiv), 2-dicyclohexylphosphine-2,4,6-triisopropyl Biphenyl (61.83 mg, 129.70 micromole, 0.2 eq.) was reacted at 70°C for 12 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1) was further subjected to high performance liquid chromatography (column: Waters Xbridge C18 150*50 mm*10 microns; mobile phase: mobile phase A: Aqueous solution of ammonium bicarbonate; mobile phase B: acetonitrile; B%: 33%-63%, 11 minutes) isolated to obtain compound 22.
MS-ESI计算值[M+H] +546,实测值546。 MS-ESI calculated [M+H] + 546, found 546.
1HNMR(400MHz,CD 3OD)δ＝8.41–8.22(m,1H),8.02–7.88(m,1H),7.60-7.48(m,1H),7.38-7.29(m,1H),7.22-7.06(m,2H),6.45-6.36(m,1H),5.03(s,2H),4.60(br s,2H),3.94(s,4H),2.82(t,J＝12.17Hz,4H),1.85(s,3H),1.81(s,3H). 1 HNMR (400MHz, CD 3 OD) δ=8.41–8.22(m,1H),8.02–7.88(m,1H),7.60-7.48(m,1H),7.38-7.29(m,1H),7.22-7.06 (m,2H),6.45-6.36(m,1H),5.03(s,2H),4.60(br s,2H),3.94(s,4H),2.82(t,J＝12.17Hz,4H),1.85 (s,3H),1.81(s,3H).
实施例23Example 23
Figure PCTCN2022129958-appb-000070
Figure PCTCN2022129958-appb-000070
第一步first step
将化合物17-3(300毫克，1.64毫摩尔，1当量)和化合物H(324.78毫克，3.28毫摩尔，2当量)溶于乙腈(10毫升)，接着向反应液中加入碳酸钾(679.2毫克，4.9毫摩尔，3当量)，在80摄氏度氮气保护下反应2小时。反应完成后，乙酸乙酯萃取(30毫升×3)，饱和食盐水(20毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩得到化合物23-1。Compound 17-3 (300 mg, 1.64 mmol, 1 eq) and compound H (324.78 mg, 3.28 mmol, 2 eq) were dissolved in acetonitrile (10 ml), and potassium carbonate (679.2 mg, 4.9 mmol, 3 equivalents), and reacted for 2 hours at 80 degrees Celsius under nitrogen protection. After the reaction was complete, the mixture was extracted with ethyl acetate (30 mL×3), washed with saturated brine (20 mL×2), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 23-1.
MS-ESI计算值[M+H] +263实测值263。 MS-ESI calculated [M+H] + 263 found 263.
第二步second step
将化合物23-1(320.00毫克，1.22毫摩尔，1当量)溶于甲醇(10毫升)，接着向反应液中加入钯碳 (50.00毫克，纯度：10％)，在25摄氏度氢气氛围(15PSI)下反应2小时。反应完成后，垫硅藻土过滤，滤液浓缩得到化合物23-2。Compound 23-1 (320.00 mg, 1.22 mmol, 1 eq) was dissolved in methanol (10 ml), then palladium on carbon (50.00 mg, purity: 10%) was added to the reaction solution, and the compound was dissolved in a hydrogen atmosphere (15PSI) at 25 degrees Celsius. The reaction was carried out for 2 hours. After the reaction was completed, it was filtered with celite, and the filtrate was concentrated to obtain compound 23-2.
MS-ESI计算值[M+H] +233,实测值233。 MS-ESI calculated [M+H] + 233, found 233.
第三步third step
将化合物23-2(300毫克，1.29毫摩尔，1.1当量)溶于四氢呋喃(10毫升)，接着向反应液中加入中间体E(370.17毫克，1.17毫摩尔，1当量)，叔丁醇钠(225.67毫克，1.68毫摩尔，2当量)，在20摄氏度氮气保护下加入三(二亚苄基丙酮)二钯(107.52毫克，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(111.75毫克，0.2当量),然后加热70摄氏度反应12小时。反应完成后过滤后得滤液，加入水(10毫升)，乙酸乙酯(30毫升×2)，有机相经饱和食盐水(10毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩。粗品经高效液相色谱(色谱柱：Waters Xbridge C18 150*50毫米*10微米；流动相A：碳酸氢铵水溶液，流动相B：乙腈；B％:20％-50％,11分钟)分离得到化合物23。Compound 23-2 (300 mg, 1.29 mmol, 1.1 eq) was dissolved in tetrahydrofuran (10 ml), then Intermediate E (370.17 mg, 1.17 mmol, 1 eq), sodium tert-butoxide ( 225.67 mg, 1.68 mmol, 2 equivalents), added tris(dibenzylideneacetone) dipalladium (107.52 mg, 0.1 equivalents), 2-dicyclohexylphosphine-2,4,6-tri Isopropyl biphenyl (111.75 mg, 0.2 equivalents), then heated at 70 degrees Celsius for 12 hours. After the reaction was completed, the filtrate was obtained after filtration, water (10 ml), ethyl acetate (30 ml × 2) were added, the organic phase was washed with saturated brine (10 ml × 2), the organic phase was dried with anhydrous sodium sulfate, filtered, concentrate. The crude product was separated by high performance liquid chromatography (column: Waters Xbridge C18 150*50 mm*10 microns; mobile phase A: aqueous ammonium bicarbonate solution, mobile phase B: acetonitrile; B%: 20%-50%, 11 minutes) to obtain Compound 23.
MS-ESI计算值[M+H] +512,实测值512。 MS-ESI calculated value [M+H] + 512, found value 512.
1H NMR(400MHz,CDCl 3)δ＝10.99(s,1H),8.55(dd,J＝4.3,8.5Hz,1H),8.05(s,1H),7.40-7.31(m,2H),7.28-7.19(m,1H),7.16-7.04(m,1H),6.44(s,1H),6.34(d,J＝8.3Hz,1H),5.12(s,2H),5.04(s,2H),4.86(s,4H),4.05(s,4H),1.85(d,J＝13.2Hz,6H). 1 H NMR (400MHz, CDCl 3 )δ=10.99(s,1H),8.55(dd,J=4.3,8.5Hz,1H),8.05(s,1H),7.40-7.31(m,2H),7.28- 7.19(m,1H),7.16-7.04(m,1H),6.44(s,1H),6.34(d,J＝8.3Hz,1H),5.12(s,2H),5.04(s,2H),4.86 (s,4H),4.05(s,4H),1.85(d,J=13.2Hz,6H).
实施例24Example 24
Figure PCTCN2022129958-appb-000071
Figure PCTCN2022129958-appb-000071
第一步first step
将化合物17-3(250.00毫克，1.37毫摩尔，1当量)溶于N,N-二甲基甲酰胺(5毫升)，接着向反应液中加入碳酸钾(566.01毫克，4.10毫摩尔，3当量)，F(212.37毫克，1.37毫摩尔，1当量)，在80摄氏度下反应2小时。反应完成后，加水(30毫升)稀释，过滤，滤饼干燥得到化合物24-1。Compound 17-3 (250.00 mg, 1.37 mmol, 1 eq.) was dissolved in N,N-dimethylformamide (5 ml), and then potassium carbonate (566.01 mg, 4.10 mmol, 3 eq. ), F (212.37 mg, 1.37 mmol, 1 equivalent), reacted at 80 degrees Celsius for 2 hours. After the reaction was completed, it was diluted with water (30 mL), filtered, and the filter cake was dried to obtain compound 24-1.
MS-ESI计算值[M+H] +283,实测值283。 MS-ESI calculated [M+H] + 283, found 283.
第二步second step
将化合物24-1(350.00毫克，1.24毫摩尔，1当量)溶于甲醇(10毫升)，接着向反应液中加入钯碳(50.00毫克，纯度：10％)，在25摄氏度氢气氛围(50PSI)下反应2小时。反应完成后，垫硅藻土过滤，滤液浓缩得到化合物24-2。Compound 24-1 (350.00 mg, 1.24 mmol, 1 eq) was dissolved in methanol (10 mL), then palladium carbon (50.00 mg, purity: 10%) was added to the reaction solution, and the compound was dissolved in a hydrogen atmosphere (50 PSI) at 25 degrees Celsius. The reaction was carried out for 2 hours. After the reaction was completed, it was filtered with celite, and the filtrate was concentrated to obtain compound 24-2.
MS-ESI计算值[M+H] +253,实测值253。 MS-ESI calculated [M+H] + 253, found 253.
第三步third step
将化合物24-2(248.97毫克，986.96微摩尔，1.2当量)，中间体E(260.00毫克，822.47微摩尔，1当量)溶于四氢呋喃(10毫升)，接着向反应液中加入叔丁醇钠(158.08毫克，1.64毫摩尔，2当量)，三(二亚苄基丙酮)二钯(75.31毫克，82.25微摩尔，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(78.42毫克，164.49微摩尔，0.2当量)，在70摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)纯化得到的粗品进一步经高效液相色谱(色谱柱：Waters Xbridge C18 150*50毫米*10微米；流动相A：碳酸氢铵水溶液；流动相B：乙腈；B％:33％-6％，11分钟)分离得到化合物24。Compound 24-2 (248.97 mg, 986.96 micromoles, 1.2 equivalents), Intermediate E (260.00 mg, 822.47 micromoles, 1 equivalents) were dissolved in tetrahydrofuran (10 ml), and sodium tert-butoxide ( 158.08 mg, 1.64 mmol, 2 equiv), tris(dibenzylideneacetone)dipalladium (75.31 mg, 82.25 micromol, 0.1 equiv), 2-dicyclohexylphosphine-2,4,6-triisopropyl Biphenyl (78.42 mg, 164.49 micromole, 0.2 eq.) was reacted at 70°C for 12 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1) is further purified by high-performance liquid chromatography (column: Waters Xbridge C18 150*50 millimeters*10 microns; mobile phase A: ammonium bicarbonate Aqueous solution; mobile phase B: acetonitrile; B%: 33%-6%, 11 minutes) isolated to obtain compound 24.
MS-ESI计算值[M+H] +532,实测值532。 MS-ESI calculated [M+H] + 532, found 532.
1HNMR(400MHz,CD 3OD)δ＝8.29-8.39(m,1H)8.00(s,1H)7.50-7.62(m,1H)7.36(br t,J＝7.76Hz,1H)7.13-7.26(m,2H)6.43-6.49(m,1H)5.04-5.12(m,2H)4.92-4.93(m,2H)4.08-4.12(m,2H)3.98-4.05(m,2H)1.88(s,3H)1.84(s,3H)1.57-1.65(m,2H). 1 HNMR (400MHz, CD 3 OD) δ = 8.29-8.39 (m, 1H) 8.00 (s, 1H) 7.50-7.62 (m, 1H) 7.36 (br t, J = 7.76Hz, 1H) 7.13-7.26 (m ,2H)6.43-6.49(m,1H)5.04-5.12(m,2H)4.92-4.93(m,2H)4.08-4.12(m,2H)3.98-4.05(m,2H)1.88(s,3H)1.84 (s,3H)1.57-1.65(m,2H).
实施例25Example 25
Figure PCTCN2022129958-appb-000072
Figure PCTCN2022129958-appb-000072
第一步first step
将化合物17-3(300毫克，1.64毫摩尔，1当量)和化合物J(536.14毫克，3.28毫摩尔，2当量)溶于乙腈(10毫升)，接着向反应液中加入碳酸钾(679.2毫克，4.9毫摩尔，3当量)，在80摄氏度氮气保护下反应2小时。反应完成后，乙酸乙酯萃取(30毫升×3)，饱和食盐水(20毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩得到化合物25-1。Compound 17-3 (300 mg, 1.64 mmol, 1 eq) and compound J (536.14 mg, 3.28 mmol, 2 eq) were dissolved in acetonitrile (10 ml), and potassium carbonate (679.2 mg, 4.9 mmol, 3 equivalents), and reacted for 2 hours at 80 degrees Celsius under nitrogen protection. After the reaction was complete, the mixture was extracted with ethyl acetate (30 mL×3), washed with saturated brine (20 mL×2), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 25-1.
MS-ESI计算值[M+H] +291,实测值291。 MS-ESI calculated value [M+H] + 291, found value 291.
第二步second step
将化合物25-1(390.00毫克，1.34毫摩尔，1当量)溶于甲醇(10毫升)，接着向反应液中加入钯碳(100.00毫克，纯度：10％)，在25摄氏度氢气氛围(15PSI)下反应2小时。反应完成后，垫硅藻土过滤，滤液浓缩得到化合物25-2。Compound 25-1 (390.00 mg, 1.34 mmol, 1 eq) was dissolved in methanol (10 ml), then palladium on carbon (100.00 mg, purity: 10%) was added to the reaction solution, and the The reaction was carried out for 2 hours. After the reaction was completed, it was filtered with celite, and the filtrate was concentrated to obtain compound 25-2.
MS-ESI计算值[M+H] +261,实测值261。 MS-ESI calculated value [M+H] + 261, found value 261.
第三步third step
将化合物25-2(300毫克，1.15毫摩尔，1.1当量)溶于四氢呋喃(10毫升)，接着向反应液中加入中间体E(331.18毫克，1.05毫摩尔，1当量)，叔丁醇钠(201.36毫克，2.10毫摩尔，2当量)，在20摄氏度氮气保护下加入三(二亚苄基丙酮)二钯(95.93毫克，0.1当量)，2-二环己基磷-2,4,6-三异丙基联苯(99.88毫克，0.2当量)，然后加热70摄氏度反应12小时。反应完成后过滤后得滤液，加入水(10毫升)，乙酸乙酯(30毫升×2)，有机相经饱和食盐水(10毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩。粗品经高效液相色谱(色谱柱：Waters Xbridge C18 150*50毫米*10微米；流动相A：碳酸氢铵水溶液，流动相B：乙腈；B％：27％-57％,11分钟)分离得到化合物25。Compound 25-2 (300 mg, 1.15 mmol, 1.1 eq) was dissolved in tetrahydrofuran (10 ml), then Intermediate E (331.18 mg, 1.05 mmol, 1 eq), sodium tert-butoxide ( 201.36 mg, 2.10 mmol, 2 equivalents), added tris(dibenzylideneacetone) dipalladium (95.93 mg, 0.1 equivalents), 2-dicyclohexylphosphine-2,4,6-tris Isopropyl biphenyl (99.88 mg, 0.2 equivalents) was then heated at 70°C for 12 hours. After the reaction was completed, the filtrate was obtained after filtration, water (10 ml), ethyl acetate (30 ml × 2) were added, the organic phase was washed with saturated brine (10 ml × 2), the organic phase was dried with anhydrous sodium sulfate, filtered, concentrate. The crude product was separated by high performance liquid chromatography (column: Waters Xbridge C18 150*50 mm*10 microns; mobile phase A: aqueous ammonium bicarbonate solution, mobile phase B: acetonitrile; B%: 27%-57%, 11 minutes) to obtain Compound 25.
MS-ESI计算值[M+H] +540,实测值540。 MS-ESI calculated value [M+H] + 540, found value 540.
1H NMR(400MHz,CD 3OD)δ＝8.35(br s,1H),7.99(s,1H),7.54(ddd,J＝1.5,7.7,14.4Hz,1H),7.30(br t,J＝6.8Hz,1H),7.18(br t,J＝7.5Hz,1H),7.11(d,J＝8.5Hz,1H),6.56(d,J＝8.6Hz,1H),5.26(s,2H),4.98-4.92(m,2H),3.96(t,J＝7.1Hz,2H),3.79-3.68(m,2H),3.51-3.42(m,2H),3.39-3.35(m,1H),2.11-1.98(m,4H),1.88(s,3H),1.84(s,3H). 1 H NMR (400MHz, CD 3 OD) δ=8.35(br s,1H),7.99(s,1H),7.54(ddd,J=1.5,7.7,14.4Hz,1H),7.30(br t,J= 6.8Hz, 1H), 7.18(br t, J=7.5Hz, 1H), 7.11(d, J=8.5Hz, 1H), 6.56(d, J=8.6Hz, 1H), 5.26(s, 2H), 4.98-4.92(m,2H),3.96(t,J＝7.1Hz,2H),3.79-3.68(m,2H),3.51-3.42(m,2H),3.39-3.35(m,1H),2.11- 1.98(m,4H),1.88(s,3H),1.84(s,3H).
实施例26Example 26
Figure PCTCN2022129958-appb-000073
Figure PCTCN2022129958-appb-000073
第一步first step
将化合物E(5克，15.82毫摩尔，1当量)溶于异丙醇(50毫升)，然后向反应液中加入化合物1-1(4.15克，20.56毫摩尔，1.3当量)和甲烷磺酸(1.52克，15.82毫摩尔，1当量)，在90摄氏度下反应12小时。反应完成后，将反应液减压浓缩，随后，加水(50毫升)，并用饱和的碳酸氢钠溶液将溶液的pH调至8，然后加入二氯甲烷(50毫升)稀释反应液，用二氯甲烷(50毫升×3)萃取，有机相经无水硫酸钠干燥，过滤，减压浓缩。粗品经高效液相色谱(色谱柱：Phenomenex luna C18 250×70毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:35％-60％，21分钟)分离得到化合物26-1。Compound E (5 g, 15.82 mmol, 1 eq) was dissolved in isopropanol (50 ml), then compound 1-1 (4.15 g, 20.56 mmol, 1.3 eq) and methanesulfonic acid ( 1.52 g, 15.82 mmol, 1 equivalent), reacted at 90 degrees Celsius for 12 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure, then, water (50 ml) was added, and the pH of the solution was adjusted to 8 with saturated sodium bicarbonate solution, then dichloromethane (50 ml) was added to dilute the reaction solution, and dichloromethane was added to Extracted with methane (50 mL×3), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by high performance liquid chromatography (chromatographic column: Phenomenex luna C18 250 × 70 mm × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 35%-60%, 21 minutes) separation Compound 26-1 was obtained.
MS-ESI计算值[M+H] +483，实测值483。 MS-ESI calculated [M+H] + 483, found 483.
第二步second step
将化合物26-1(200毫克，415.19微摩尔，1当量)溶于二氧六环(20毫升)，接着向反应液中加入化 合物K(115.27毫克，622.78微摩尔，1.5当量)，叔丁醇钠(119.70毫克，1.25毫摩尔，3当量)，4,5-双(二苯基磷)-9,9-二甲基氧杂蒽(48.05毫克，83.04微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(38.02毫克，41.52微摩尔，0.1当量)，置换三次氮气，并在氮气氛围下在100摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)纯化得到的粗品经高效液相色谱(色谱柱：Phenomenex Luna C18 150×25毫米×10微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％:4％-34％，10分钟)分离得到化合物26。Compound 26-1 (200 mg, 415.19 micromoles, 1 equivalent) was dissolved in dioxane (20 milliliters), then compound K (115.27 mg, 622.78 micromoles, 1.5 equivalents), tert-butyl alcohol were added to the reaction solution Sodium (119.70 mg, 1.25 mmol, 3 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (48.05 mg, 83.04 micromol, 0.2 equiv), tris(diphenyl Benzylideneacetone) dipalladium (38.02 mg, 41.52 micromole, 0.1 equiv), nitrogen was replaced three times, and reacted at 100° C. for 12 hours under nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1) was purified by high performance liquid chromatography (column: Phenomenex Luna C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 4%-34%, 10 minutes) isolated to obtain compound 26.
MS-ESI计算值[M+H] +513,实测值513. MS-ESI calculated value [M+H] + 513, measured value 513.
1H NMR(400MHz,CD 3OD)δ＝8.39-8.29(m,1H),8.16-8.08(m,1H),7.72-7.61(m,1H),7.61-7.51(m,1H),7.49-7.41(m,1H),7.36-7.27(m,1H),6.91-6.80(m,1H),6.31-6.21(m,1H),4.08-3.96(m,4H),3.91-3.82(m,4H),3.70-3.63(m,3H),2.68-2.57(m,3H),1.90-1.81(m,6H)。 1 H NMR (400MHz, CD 3 OD) δ=8.39-8.29(m,1H),8.16-8.08(m,1H),7.72-7.61(m,1H),7.61-7.51(m,1H),7.49- 7.41(m,1H),7.36-7.27(m,1H),6.91-6.80(m,1H),6.31-6.21(m,1H),4.08-3.96(m,4H),3.91-3.82(m,4H ), 3.70-3.63 (m, 3H), 2.68-2.57 (m, 3H), 1.90-1.81 (m, 6H).
实施例27Example 27
Figure PCTCN2022129958-appb-000074
Figure PCTCN2022129958-appb-000074
第一步first step
将化合物7-2(200毫克，555.986微摩尔，1当量)溶于二氧六环(3毫升)，接着向反应液中加入化合物K(154.33毫克，833.79微摩尔，1.5当量)，碳酸铯(724.44毫克，2.22毫摩尔，4当量)，四氟硼酸三叔丁基膦(32.25毫克，111.17微摩尔，0.2当量)和醋酸钯(12.48毫克，55.59微摩尔，0.1当量)，置换三次氮气，并在氮气氛围下在100摄氏度下反应3小时。反应完成后，将反应液过滤，减压浓缩。粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝5：1)纯化得到化合物27-1。Compound 7-2 (200 mg, 555.986 micromoles, 1 equivalent) was dissolved in dioxane (3 ml), and then compound K (154.33 mg, 833.79 micromoles, 1.5 equivalents), cesium carbonate ( 724.44 mg, 2.22 mmol, 4 equivalents), tri-tert-butylphosphine tetrafluoroborate (32.25 mg, 111.17 micromoles, 0.2 equivalents) and palladium acetate (12.48 mg, 55.59 micromoles, 0.1 equivalents), three nitrogen displacements, and The reaction was carried out at 100° C. for 3 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered and concentrated under reduced pressure. The crude product was purified by silica gel thin-layer chromatography (dichloromethane:methanol=5:1) to obtain compound 27-1.
MS-ESI计算值[M+H] +436，实测值436。 MS-ESI calculated [M+H] + 436, found 436.
第二步second step
将化合物27-1溶于二氯甲烷(1毫升)和三氟乙酸(1毫升)的混合溶液中，在室温下搅拌0.5小时。反应完成后，将反应液减压浓缩，得到化合物27-2(三氟乙酸盐)Compound 27-1 was dissolved in a mixed solution of dichloromethane (1 mL) and trifluoroacetic acid (1 mL), and stirred at room temperature for 0.5 hr. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain compound 27-2 (trifluoroacetate)
MS-ESI计算值[M+H] +236，实测值236。 MS-ESI calculated [M+H] +236 , found 236.
第三步third step
将化合物27-2(100毫克，144.64微摩尔，三氟乙酸盐，1当量)溶于四氢呋喃(10毫升)，接着向反应液中加入化合物E(59.44毫克，188.03微摩尔，1.3当量)，叔丁醇钠(69.50毫克，723.19微摩尔，5当 量)，2-二环己基磷-2,4,6-三异丙基联苯(13.79毫克，28.93微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(26.49毫克，28.93微摩尔，0.2当量)，置换三次氮气，并在氮气氛围下在70摄氏度下反应2小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)纯化得到的粗品经高效液相色谱(色谱柱：Phenomenex Luna C18 150×25毫米×10微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％:1％-30％，10分钟)分离得到化合物27。Compound 27-2 (100 mg, 144.64 micromoles, trifluoroacetate salt, 1 equivalent) was dissolved in tetrahydrofuran (10 ml), and then compound E (59.44 mg, 188.03 micromoles, 1.3 equivalents) was added to the reaction solution, Sodium tert-butoxide (69.50 mg, 723.19 micromoles, 5 equivalents), 2-dicyclohexylphosphonium-2,4,6-triisopropylbiphenyl (13.79 mg, 28.93 micromoles, 0.2 equivalents), tri(di (benzylideneacetone) dipalladium (26.49 mg, 28.93 micromole, 0.2 eq), nitrogen was replaced three times, and reacted at 70° C. for 2 hours under nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1) was purified by high performance liquid chromatography (column: Phenomenex Luna C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 1%-30%, 10 minutes) isolated to obtain compound 27.
MS-ESI计算值[M+H] +515,实测值515. MS-ESI calculated value [M+H] + 515, measured value 515.
1H NMR(400MHz,CD 3OD)δ＝8.39-8.30(m,1H),8.19(s,1H),8.02(s,1H),7.57(dd,J＝7.8,13.7Hz,1H),7.48-7.42(m,1H),7.24-7.18(m,1H),4.24(s,4H),3.94(s,3H),3.72(s,4H),2.51(s,3H),1.88(s,3H),1.85(s,3H)。 1 H NMR (400MHz, CD 3 OD) δ = 8.39-8.30 (m, 1H), 8.19 (s, 1H), 8.02 (s, 1H), 7.57 (dd, J = 7.8, 13.7Hz, 1H), 7.48 -7.42(m,1H),7.24-7.18(m,1H),4.24(s,4H),3.94(s,3H),3.72(s,4H),2.51(s,3H),1.88(s,3H ), 1.85(s,3H).
实施例28Example 28
Figure PCTCN2022129958-appb-000075
Figure PCTCN2022129958-appb-000075
第一步first step
将化合物28-1(1克，4.00毫摩尔，1当量)溶于乙醇(15毫升)和水(8毫升)中，接着向反应液中加入氯化铵(1.71克，32.00毫摩尔，8当量)和铁粉(1.34克，24.00毫摩尔，6当量)，在70摄氏度下反应2小时。反应完成后，将反应液减压浓缩，然后加入二氯甲烷(50毫升)稀释反应液，用二氯甲烷(50毫升×3)萃取，有机相经无水硫酸钠干燥，过滤，减压浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝20：1到8：1)分离得到化合物28-2。Compound 28-1 (1 g, 4.00 mmol, 1 eq) was dissolved in ethanol (15 ml) and water (8 ml), then ammonium chloride (1.71 g, 32.00 mmol, 8 eq) was added to the reaction solution ) and iron powder (1.34 g, 24.00 mmol, 6 equivalents), reacted at 70 degrees Celsius for 2 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure, then dichloromethane (50 ml) was added to dilute the reaction solution, extracted with dichloromethane (50 ml×3), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure . The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate=20:1 to 8:1) to obtain compound 28-2.
MS-ESI计算值[M+H] +220，实测值220。 MS-ESI calculated [M+H] +220 , found 220.
第二步second step
将化合物28-2(400毫克，1.82毫摩尔，1当量)溶于异丙醇(10毫升)，然后向反应液中加入化合物E(689.60毫克，2.18毫摩尔，1.2当量)和甲基磺酸(174.71毫克，1.82毫摩尔，129.41微升，1当量)，在90摄氏度下反应12小时。反应完成后，将反应液减压浓缩，随后，加水(50毫升)，并用饱和的碳酸氢钠溶液将溶液的pH调至8，然后加入二氯甲烷(30毫升)稀释反应液，用二氯甲烷(30毫升×3)萃取，有机相经无水硫酸钠干燥，过滤，减压浓缩。粗品经高效液相色谱(色谱柱：Phenomenex luna C8 250×50毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：40％-70％，20分钟)分离得到化合物28-3。Compound 28-2 (400 mg, 1.82 mmol, 1 eq) was dissolved in isopropanol (10 ml), then compound E (689.60 mg, 2.18 mmol, 1.2 eq) and methanesulfonic acid were added to the reaction solution (174.71 mg, 1.82 mmol, 129.41 microliters, 1 equiv), reacted at 90 degrees Celsius for 12 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure, then, water (50 ml) was added, and the pH of the solution was adjusted to 8 with saturated sodium bicarbonate solution, then dichloromethane (30 ml) was added to dilute the reaction solution, and dichloromethane was added to Extracted with methane (30 mL×3), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by high performance liquid chromatography (chromatographic column: Phenomenex luna C8 250 × 50 mm × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 40%-70%, 20 minutes) separation Compound 28-3 was obtained.
MS-ESI计算值[M+H] +501，实测值501。 MS-ESI calculated [M+H] + 501, found 501.
第三步third step
将化合物28-3(200毫克，400.24微摩尔，1当量)溶于二氧六环(12毫升)，接着向反应液中加入化合物F(124.53毫克，800.48微摩尔，2当量)，叔丁醇钠(153.86毫克，1.60毫摩尔，4当量)，4,5-双(二苯基磷)-9,9-二甲基氧杂蒽(46.32毫克，80.05微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(73.30毫克，80.05微摩尔，0.2当量)，置换三次氮气，并在氮气氛围下在110摄氏度下反应5小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)纯化得到的粗品经高效液相色谱(色谱柱：Phenomenex Luna C18 150×25毫米×10微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％：51％-81％，10分钟)分离得到化合物28。Compound 28-3 (200 mg, 400.24 micromoles, 1 eq) was dissolved in dioxane (12 ml), then compound F (124.53 mg, 800.48 micromoles, 2 eqs), tert-butyl alcohol were added to the reaction solution Sodium (153.86 mg, 1.60 mmol, 4 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (46.32 mg, 80.05 micromol, 0.2 equiv), tris(diphenyl Benzylideneacetone) dipalladium (73.30 mg, 80.05 micromole, 0.2 eq), nitrogen was replaced three times, and reacted at 110°C for 5 hours under nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product purified by silica gel thin-layer chromatography (dichloromethane:methanol=10:1) was purified by high performance liquid chromatography (column: Phenomenex Luna C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 51%-81%, 10 minutes) isolated to obtain compound 28.
MS-ESI计算值[M+H] +538,实测值538. MS-ESI calculated value [M+H] + 538, measured value 538.
1H NMR(400MHz,CD 3OD)δ＝8.29(dd,J＝4.5,8.3Hz,1H),8.17(br s,1H),7.74-7.59(m,2H),7.41(dd,J＝2.9,11.2Hz,1H),7.38-7.30(m,1H),6.03(dd,J＝3.1,10.1Hz,1H),4.13(d,J＝7.8Hz,2H),4.00(br d,J＝7.9Hz,2H),3.68(s,3H),1.87(d,J＝13.6Hz,6H),1.57(t,J＝8.6Hz,2H)。 1 H NMR (400MHz, CD 3 OD) δ = 8.29 (dd, J = 4.5, 8.3 Hz, 1H), 8.17 (br s, 1H), 7.74-7.59 (m, 2H), 7.41 (dd, J = 2.9 ,11.2Hz,1H),7.38-7.30(m,1H),6.03(dd,J=3.1,10.1Hz,1H),4.13(d,J=7.8Hz,2H),4.00(br d,J=7.9 Hz, 2H), 3.68 (s, 3H), 1.87 (d, J=13.6Hz, 6H), 1.57 (t, J=8.6Hz, 2H).
实施例29Example 29
Figure PCTCN2022129958-appb-000076
Figure PCTCN2022129958-appb-000076
第一步first step
将化合物29-1(4.5克，19.07毫摩尔1当量)，碘甲烷(6.77克，47.67毫摩尔，2.5当量)，碳酸铯(15.53克，47.67毫摩尔，2.5当量)溶于丙酮(100毫升)中，接着在70摄氏度氮气保护下反应3小时。反应完毕后，向反应液中加入水(100毫升)稀释，用乙酸乙酯萃取(100毫升×2)，有机相用饱和的食盐水(200毫升)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100:1-80:1)分离纯化得到化合物29-2。Compound 29-1 (4.5 g, 19.07 mmol 1 eq), iodomethane (6.77 g, 47.67 mmol, 2.5 eq), cesium carbonate (15.53 g, 47.67 mmol, 2.5 eq) were dissolved in acetone (100 ml) , followed by reaction at 70°C for 3 hours under nitrogen protection. After the reaction was completed, water (100 ml) was added to the reaction solution for dilution, extracted with ethyl acetate (100 ml × 2), the organic phase was washed with saturated brine (200 ml), dried over anhydrous sodium sulfate, filtered, and concentrated The residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-80:1) to obtain compound 29-2.
第二步second step
将化合物29-2(3.85克，15.4毫摩尔，1当量)溶于乙醇(40毫升)和水(20毫升)中，加入铁粉(5.16克，92.39毫摩尔，6当量)，氯化铵(6.59克，123.19毫摩尔，8当量)，在70摄氏下反应2小时。反应完毕后，向反应液中加入水(100毫升)稀释，用乙酸乙酯萃取(100毫升×2)，有机相用饱和的食盐水(200毫升)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100:1-10:1)分离纯化得到化合物29-3。Compound 29-2 (3.85 g, 15.4 mmol, 1 eq) was dissolved in ethanol (40 ml) and water (20 ml), iron powder (5.16 g, 92.39 mmol, 6 eq), ammonium chloride ( 6.59 g, 123.19 mmol, 8 equivalents), reacted at 70 degrees Celsius for 2 hours. After the reaction was completed, water (100 ml) was added to the reaction solution for dilution, extracted with ethyl acetate (100 ml × 2), the organic phase was washed with saturated brine (200 ml), dried over anhydrous sodium sulfate, filtered, and concentrated The residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-10:1) to obtain compound 29-3.
第三步third step
将化合物29-3(2.9克，13.18毫摩尔，1当量)溶于异丙醇(30毫升)中，再加入化合物E(5克，15.82毫摩尔，1.2当量)，甲烷磺酸(1.27克，13.18毫摩尔，1当量)，接着在90摄氏度下反应12小时，反应完毕后，将反应混合物过滤浓缩。过滤，浓缩的残渣经制备高效液相色谱法(色谱柱:Phenomenex luna C18 250×70毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；45％-75％，20分钟)分离得到化合物29-4。Compound 29-3 (2.9 g, 13.18 mmol, 1 eq) was dissolved in isopropanol (30 ml), and compound E (5 g, 15.82 mmol, 1.2 eq), methanesulfonic acid (1.27 g, 13.18 mmoles, 1 equivalent), then reacted at 90 degrees Celsius for 12 hours, after the reaction was completed, the reaction mixture was concentrated by filtration. Filtration, the concentrated residue was subjected to preparative high performance liquid chromatography (chromatographic column: Phenomenex luna C18 250 × 70 mm × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; 45%-75%, 20 minutes) to isolate compound 29-4.
MS-ESI计算值[M+H] +501，实测值501。 MS-ESI calculated [M+H] + 501, found 501.
1HNMR(400MHz,CDCl 3)δ＝10.97(s,1H),8.52(dd,J＝4.3,8.4Hz,1H),8.26(dd,J＝5.8,9.2Hz,1H),8.12(s,1H),7.50(t,J＝7.9Hz,1H),7.37-7.27(m,2H),7.15(dt,J＝1.5,7.4Hz,1H),6.87(dd,J＝8.1,9.1Hz,1H),3.90(s,3H),1.84(d,J＝13.1Hz,6H)。 1 HNMR (400MHz, CDCl 3 )δ=10.97(s,1H),8.52(dd,J=4.3,8.4Hz,1H),8.26(dd,J=5.8,9.2Hz,1H),8.12(s,1H ),7.50(t,J=7.9Hz,1H),7.37-7.27(m,2H),7.15(dt,J=1.5,7.4Hz,1H),6.87(dd,J=8.1,9.1Hz,1H) , 3.90 (s, 3H), 1.84 (d, J=13.1Hz, 6H).
第四步the fourth step
将化合物29-4(250毫克，500.3微摩尔，1当量)，化合物F(116.75毫克，750.45毫摩尔，1.5当量)，叔丁醇钠(144.24毫克，1.5毫摩尔，3当量)，三(二亚苄基丙酮)二钯(45.81毫克，50.03微摩尔，0.1当量)，4,5-双(二苯基磷)-9,9-二甲基氧杂蒽(57.9毫克，100.06微摩尔，0.2当量)，溶于二氧六环(10毫升)中接着在115摄氏度氮气保护下反应12小时，反应完毕后，向反应液中加入水(50毫升)稀释，用乙酸乙酯萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥。过滤，浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)和制备高效液相色谱法(色谱柱:Phenomenex Synergi C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；44％-74％，10分钟)分离得到化合物29。Compound 29-4 (250 mg, 500.3 micromoles, 1 equivalent), compound F (116.75 mg, 750.45 mmoles, 1.5 equivalents), sodium tert-butoxide (144.24 mg, 1.5 mmoles, 3 equivalents), three (di Benzylideneacetone) dipalladium (45.81 mg, 50.03 micromoles, 0.1 eq), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (57.9 mg, 100.06 micromoles, 0.2 equivalent), be dissolved in dioxane (10 milliliters) and then react at 115 degrees centigrade under nitrogen protection for 12 hours. After the completion of the reaction, add water (50 milliliters) to the reaction solution for dilution, and extract with ethyl acetate (50 milliliters× 2), the organic phase was washed with saturated brine (50 ml), and dried over anhydrous sodium sulfate. Filtration, the concentrated residue was subjected to silica gel thin-layer chromatography (dichloromethane:methanol=10:1) and preparative high-performance liquid chromatography (chromatographic column: Phenomenex Synergi C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A : aqueous formic acid; mobile phase B: acetonitrile; 44%-74%, 10 minutes) isolated to obtain compound 29.
MS-ESI计算值[M+H] +538，实测值538。 MS-ESI calculated [M+H] + 538, found 538.
1H NMR(400MHz,CD 3OD)δ＝8.30(dd,J＝4.4,8.2Hz,1H),8.07(s,1H),7.90-7.72(m,1H),7.67-7.57(m,1H),7.56-7.48(m,1H),7.33-7.25(m,1H),7.25-7.19(m,1H),6.96(br d,J＝4.0Hz,1H),6.67(dd,J＝9.2,12.6Hz,1H),6.14(d,J＝7.7Hz,1H),4.58(s,1H),4.41-4.10(m,4H),3.63(s,3H),1.84(d,J＝13.4Hz,6H),1.64-1.57(m,1H),1.51(t,J＝8.6Hz,2H),1.40-1.26(m,1H)。 1 H NMR (400MHz, CD 3 OD) δ=8.30(dd,J=4.4,8.2Hz,1H),8.07(s,1H),7.90-7.72(m,1H),7.67-7.57(m,1H) ,7.56-7.48(m,1H),7.33-7.25(m,1H),7.25-7.19(m,1H),6.96(br d,J=4.0Hz,1H),6.67(dd,J=9.2,12.6 Hz, 1H), 6.14(d, J=7.7Hz, 1H), 4.58(s, 1H), 4.41-4.10(m, 4H), 3.63(s, 3H), 1.84(d, J=13.4Hz, 6H ), 1.64-1.57 (m, 1H), 1.51 (t, J=8.6Hz, 2H), 1.40-1.26 (m, 1H).
实施例30Example 30
Figure PCTCN2022129958-appb-000077
Figure PCTCN2022129958-appb-000077
第一步first step
将化合物28-3(180毫克，360.22微摩尔，1当量)，化合物A(88.42毫克，540.32微摩尔，1.5当量)，叔丁醇钠(103.85，1.08毫摩尔，3当量)，三(二亚苄基丙酮)二钯(32.99毫克，36.02微摩尔，0.1当量)，4,5-双(二苯基磷)-9,9-二甲基氧杂蒽(41.69毫克，72.04微摩尔，0.2当量)溶于二氧六环(6毫升)中，然后在100摄氏度氮气保护下反应5小时。反应完毕后，向反应液中加入水(50毫升)，用乙酸乙酯(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经制备高效液相色谱法(色谱柱:Phenomenex luna C18 150×25毫米×10微米；流动相：流动相A：甲酸 水溶液；流动相B：乙腈；50％-80％，10分钟)分离得到化合物30。Compound 28-3 (180 mg, 360.22 micromoles, 1 equivalent), compound A (88.42 mg, 540.32 micromoles, 1.5 equivalents), sodium tert-butoxide (103.85, 1.08 mmoles, 3 equivalents), three (di Benzylacetone) dipalladium (32.99 mg, 36.02 micromoles, 0.1 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (41.69 mg, 72.04 micromoles, 0.2 equiv ) was dissolved in dioxane (6 milliliters), and then reacted for 5 hours at 100 degrees centigrade under nitrogen protection. After the reaction was completed, water (50 ml) was added to the reaction solution, ethyl acetate (50 ml × 2), the organic phase was washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the concentrated residue was The compound was separated by preparative high-performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 25 millimeters × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; 50%-80%, 10 minutes) 30.
MS-ESI计算值[M+H] +546，实测值546。 MS-ESI calculated [M+H] + 546, found 546.
1HNMR(400MHz,CD 3OD)δ＝8.27(dd,J＝4.3,8.0Hz,1H),8.13(s,1H),7.60(s,2H),7.31(br d,J＝2.8Hz,2H),5.99-5.78(m,1H),3.88-3.78(m,5H),3.62(s,3H),3.23(s,3H),2.57-2.47(m,2H),2.13-2.05(m,2H),1.84(d,J＝13.6Hz,6H)。 1 HNMR (400MHz, CD 3 OD) δ = 8.27 (dd, J = 4.3, 8.0Hz, 1H), 8.13 (s, 1H), 7.60 (s, 2H), 7.31 (br d, J = 2.8Hz, 2H ),5.99-5.78(m,1H),3.88-3.78(m,5H),3.62(s,3H),3.23(s,3H),2.57-2.47(m,2H),2.13-2.05(m,2H ), 1.84 (d, J=13.6Hz, 6H).
实施例31Example 31
Figure PCTCN2022129958-appb-000078
Figure PCTCN2022129958-appb-000078
第一步first step
将化合物28-3(400毫克，800.48微摩尔，1当量)溶于二氧六环(30毫升)，接着向反应液中加入化合物D(143.60毫克，1.20毫微摩尔，1.5当量)，叔丁醇钠(307.70毫克，3.20毫摩尔，4当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(92.63毫克，160.10微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(73.30毫克，80.05微摩尔，0.1当量)，置换三次氮气，并在氮气氛围下在100摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝50：1到二氯甲烷：甲醇＝20：1)纯化得到的粗品经高效液相色谱(色谱柱：Phenomenex Luna C18 150×25毫米×10微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％:54％-84％，10分钟)分离得到化合物31。Compound 28-3 (400 mg, 800.48 micromoles, 1 equiv) was dissolved in dioxane (30 milliliters), then compound D (143.60 mg, 1.20 nanomoles, 1.5 equiv), tert-butyl Sodium alkoxide (307.70 mg, 3.20 mmol, 4 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (92.63 mg, 160.10 micromol, 0.2 equiv), tris( Dibenzylideneacetone) dipalladium (73.30 mg, 80.05 micromole, 0.1 equiv), nitrogen was replaced three times, and reacted at 100° C. for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate=50:1 to dichloromethane:methanol=20:1) and the crude product obtained was purified by high performance liquid chromatography (chromatographic column: Phenomenex Luna C18 150×25 mm×10 Micron; mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 54%-84%, 10 minutes) isolated to obtain compound 31.
MS-ESI计算值[M+H] +502,实测值502. MS-ESI calculated value [M+H] + 502, measured value 502.
1HNMR(400MHz,CD 3OD)δ＝8.28(dd,J＝4.4,8.1Hz,1H),8.15(s,1H),7.70-7.58(m,2H),7.36-7.29(m,2H),5.94(dd,J＝2.9,10.5Hz,1H),3.99(s,4H),3.64(s,3H),1.85(d,J＝13.5Hz,6H),0.67(s,4H)。 1 HNMR (400MHz, CD 3 OD) δ = 8.28 (dd, J = 4.4, 8.1Hz, 1H), 8.15 (s, 1H), 7.70-7.58 (m, 2H), 7.36-7.29 (m, 2H), 5.94 (dd, J = 2.9, 10.5Hz, 1H), 3.99 (s, 4H), 3.64 (s, 3H), 1.85 (d, J = 13.5Hz, 6H), 0.67 (s, 4H).
实施例32Example 32
Figure PCTCN2022129958-appb-000079
Figure PCTCN2022129958-appb-000079
第一步first step
将化合物29-4(190毫克，442.98微摩尔，1当量)溶于二氧六环(15毫升)，加入化合物A(93.33毫克，570.34微摩尔，1.5当量)，叔丁醇钠(109.62毫克，1.14毫摩尔，3当量)，三(二亚苄基丙酮)二钯(34.82毫克，38.02微摩尔，0.1当量)和4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(44.00毫克，76.05微摩尔，0.2当量)。用氮气置换三次，反应液在氮气保护下升温到110摄氏度，反应16小时。反应完毕后，反应液过滤，浓缩。粗品经高效液相色谱(色谱柱：Waters Xbridge 150×25毫米×5微米；流动相：流动相A：碳酸氢铵水溶液；流动相B：乙腈；B％:45％-75％，10分钟)分离得到化合物32。Compound 29-4 (190 mg, 442.98 micromoles, 1 equivalent) was dissolved in dioxane (15 ml), and compound A (93.33 mg, 570.34 micromoles, 1.5 equivalents), sodium tert-butoxide (109.62 mg, 1.14 mmol, 3 equiv), tris(dibenzylideneacetone)dipalladium (34.82 mg, 38.02 micromol, 0.1 equiv) and 4,5-bis(diphenylphosphine)-9,9-dimethyloxy Xanthene (44.00 mg, 76.05 micromol, 0.2 equiv). Replaced with nitrogen three times, the temperature of the reaction solution was raised to 110 degrees Celsius under the protection of nitrogen, and reacted for 16 hours. After the reaction was completed, the reaction solution was filtered and concentrated. The crude product was subjected to high-performance liquid chromatography (column: Waters Xbridge 150 × 25 mm × 5 microns; mobile phase: mobile phase A: aqueous ammonium bicarbonate solution; mobile phase B: acetonitrile; B%: 45%-75%, 10 minutes) Compound 32 was isolated.
MS-ESI计算值[M+H] +546，实测值546。 MS-ESI calculated [M+H] + 546, found 546.
1H NMR(400MHz,CD 3OD)δ＝8.35-8.24(m,1H),8.07(s,1H),7.61(dd,J＝8.0,14.0Hz,1H),7.56-7.46(m,1H),7.30-7.23(m,1H),7.18(dd,J＝5.3,9.1Hz,1H),6.62(dd,J＝9.1,12.6Hz,1H),4.10-4.06(m,2H),4.04-4.00(m,2H),3.86-3.79(m,1H),3.60(s,3H),3.22(s,3H),2.54-2.45(m,2H),2.10-2.00(m,2H),1.86(s,3H),1.82(s,3H)。 1 H NMR (400MHz, CD 3 OD) δ = 8.35-8.24 (m, 1H), 8.07 (s, 1H), 7.61 (dd, J = 8.0, 14.0Hz, 1H), 7.56-7.46 (m, 1H) ,7.30-7.23(m,1H),7.18(dd,J=5.3,9.1Hz,1H),6.62(dd,J=9.1,12.6Hz,1H),4.10-4.06(m,2H),4.04-4.00 (m,2H),3.86-3.79(m,1H),3.60(s,3H),3.22(s,3H),2.54-2.45(m,2H),2.10-2.00(m,2H),1.86(s ,3H), 1.82(s,3H).
实施例33Example 33
Figure PCTCN2022129958-appb-000080
Figure PCTCN2022129958-appb-000080
第一步first step
将化合物29-4(300毫克，600.36微摩尔，1当量)溶于二氧六环(20毫升)，加入化合物D(107.70毫克，900.54微摩尔，1.5当量)，碳酸铯(586.83毫克，1.8毫摩尔，3当量)，三(二亚苄基丙酮)二钯(54.98毫克，60.04微摩尔，0.1当量)和4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(69.48毫克，120.07微摩尔，0.2当量)。用氮气置换三次，反应液在氮气保护下升温到115摄氏度，反应16小时。反应完毕后，反应液过滤，浓缩。粗品经高效液相色谱(色谱柱：Waters Xbridge 150×25毫米×5微米；流动相：流动相A：碳酸氢铵水溶液；流动相B：乙腈；B％:51％-81％，8分钟)分离得到化合物33。Compound 29-4 (300 mg, 600.36 micromoles, 1 equivalent) was dissolved in dioxane (20 ml), compound D (107.70 mg, 900.54 micromoles, 1.5 equivalents), cesium carbonate (586.83 mg, 1.8 mg mol, 3 equiv), tris(dibenzylideneacetone)dipalladium (54.98 mg, 60.04 micromol, 0.1 equiv), and 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (69.48 mg, 120.07 micromol, 0.2 equiv). Replaced with nitrogen three times, the temperature of the reaction solution was raised to 115 degrees Celsius under the protection of nitrogen, and reacted for 16 hours. After the reaction was completed, the reaction solution was filtered and concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Waters Xbridge 150 × 25 mm × 5 microns; mobile phase: mobile phase A: aqueous ammonium bicarbonate solution; mobile phase B: acetonitrile; B%: 51%-81%, 8 minutes) Compound 33 was isolated.
MS-ESI计算值[M+H] +502，实测值502。 MS-ESI calculated [M+H] + 502, found 502.
1H NMR(400MHz,CD 3OD)δ＝8.38-8.24(m,1H),8.07(s,1H),7.65-7.57(m,1H),7.57-7.50(m,1H),7.31-7.25(m,1H),7.22-7.14(m,1H),6.68-6.58(m,1H),4.24-4.14(m,4H),3.61(s,3H),1.86(s,3H),1.82(s,3H),0.63(s,4H)。 1 H NMR (400MHz, CD 3 OD) δ = 8.38-8.24 (m, 1H), 8.07 (s, 1H), 7.65-7.57 (m, 1H), 7.57-7.50 (m, 1H), 7.31-7.25 ( m,1H),7.22-7.14(m,1H),6.68-6.58(m,1H),4.24-4.14(m,4H),3.61(s,3H),1.86(s,3H),1.82(s, 3H), 0.63(s, 4H).
实施例34Example 34
Figure PCTCN2022129958-appb-000081
Figure PCTCN2022129958-appb-000081
第一步first step
将化合物29-1(300毫克，1.27毫摩尔，1当量)，氢氧化钾(855.87毫克，15.25毫摩尔，12当量)，(溴二氟甲基)三甲基硅烷(774.55毫克，3.81毫摩尔，3当量)溶于乙腈(4毫升)和水(1毫升)中，接着在25摄氏度氮气保护下反应1小时。反应完毕后，反应液过滤，滤液中加入水(50毫升)稀 释，用乙酸乙酯萃取(50毫升×2)，有机相用饱和的食盐水(100毫升)洗涤，无水硫酸钠干燥，过滤，滤液浓缩得到粗品化合物34-1。Compound 29-1 (300 mg, 1.27 mmol, 1 equivalent), potassium hydroxide (855.87 mg, 15.25 mmol, 12 equivalents), (bromodifluoromethyl) trimethylsilane (774.55 mg, 3.81 mmol , 3 equivalents) were dissolved in acetonitrile (4 mL) and water (1 mL), followed by reaction at 25 °C for 1 hour under nitrogen protection. After the reaction was completed, the reaction solution was filtered, the filtrate was diluted with water (50 ml), extracted with ethyl acetate (50 ml × 2), the organic phase was washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, and filtered , the filtrate was concentrated to obtain crude compound 34-1.
第二步second step
将化合物34-1(520毫克，1.82毫摩尔，1当量)溶于乙醇(6毫升)和水(3毫升)中，然后加入铁粉(609.27毫克，10.91毫摩尔，6当量)，氯化铵(778.03毫克，14.55毫摩尔，8当量)，在70摄氏下反应2小时。反应完毕后，反应液过滤，滤液中加入水(50毫升)稀释，用二氯甲烷萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-10；1)分离纯化得到化合物34-2。Compound 34-1 (520 mg, 1.82 mmol, 1 eq) was dissolved in ethanol (6 mL) and water (3 mL), then iron powder (609.27 mg, 10.91 mmol, 6 eq), ammonium chloride were added (778.03 mg, 14.55 mmol, 8 equivalents), reacted at 70°C for 2 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was diluted with water (50 ml), extracted with dichloromethane (50 ml × 2), the organic phase was washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, and filtered , the concentrated residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100: 1-10; 1) to obtain compound 34-2.
第三步third step
将化合物34-2(370毫克，1.45毫摩尔，1当量)溶于异丙醇(6毫升)中，再加入化合物E(456.86毫克，1.45毫摩尔，1当量)，甲烷磺酸(138.89毫克，1.45毫摩尔，1当量)，接着在90摄氏度下反应12小时，反应完毕后，将反应混合物过滤，滤液浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-0：1)分离纯化得到化合物34-3。Compound 34-2 (370 mg, 1.45 mmol, 1 eq) was dissolved in isopropanol (6 ml), and compound E (456.86 mg, 1.45 mmol, 1 eq), methanesulfonic acid (138.89 mg, 1.45 mmoles, 1 equivalent), then reacted at 90 degrees Celsius for 12 hours, after the reaction was completed, the reaction mixture was filtered, and the residue concentrated from the filtrate was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=100:1-0:1 ) separation and purification to obtain compound 34-3.
MS-ESI计算值[M+H] +536.9，实测值536.9。 MS-ESI calculated [M+H] + 536.9, found 536.9.
1HNMR(400MHz,CDCl 3)δ＝12.36(br s,1H),10.97(br s,1H),8.03(br dd,J＝4.2,7.9Hz,1H),7.82(s,1H),7.43(br dd,J＝5.5,8.8Hz,1H),7.33-7.25(m,3H),7.13(t,J＝8.1Hz,1H),6.82-6.33(m,1H),1.89-1.81(m,6H). 1 HNMR (400MHz, CDCl 3 ) δ=12.36(br s,1H),10.97(br s,1H),8.03(br dd,J=4.2,7.9Hz,1H),7.82(s,1H),7.43( br dd,J=5.5,8.8Hz,1H),7.33-7.25(m,3H),7.13(t,J=8.1Hz,1H),6.82-6.33(m,1H),1.89-1.81(m,6H ).
第四步the fourth step
将化合物34-3(250毫克，466.70微摩尔，1当量)，化合物F(108.91毫克，700.04毫摩尔，1.5当量)，叔丁醇钠(134.55毫克，1.40毫摩尔，3当量)，三(二亚苄基丙酮)二钯(42.74毫克，46.67微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(54.01毫克，93.34微摩尔，0.2当量)，溶于二氧六环(10毫升)中接着在115摄氏度氮气保护下反应12小时，反应完毕后，反应液过滤，滤液中加入水(50毫升)稀释，用乙酸乙酯萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥。过滤，浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)和制备高效液相色谱法(色谱柱:Phenomenex Synergi C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；35％-65％，10分钟)分离得到化合物34。Compound 34-3 (250 mg, 466.70 micromole, 1 equivalent), compound F (108.91 mg, 700.04 mmol, 1.5 equivalent), sodium tert-butoxide (134.55 mg, 1.40 mmol, 3 equivalents), three (di Benzylideneacetone) dipalladium (42.74 mg, 46.67 micromoles, 0.1 eq), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (54.01 mg, 93.34 micromoles, 0.2 equivalent), be dissolved in dioxane (10 milliliters) and then reacted under nitrogen protection at 115 degrees Celsius for 12 hours. After the reaction was completed, the reaction solution was filtered, and water (50 milliliters) was added to the filtrate for dilution, and extracted with ethyl acetate (50 milliliters) mL×2), the organic phase was washed with saturated brine (50 mL), and dried over anhydrous sodium sulfate. Filtration, the concentrated residue was subjected to silica gel thin-layer chromatography (dichloromethane:methanol=10:1) and preparative high-performance liquid chromatography (chromatographic column: Phenomenex Synergi C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A : aqueous formic acid; mobile phase B: acetonitrile; 35%-65%, 10 minutes) isolated to obtain compound 34.
MS-ESI计算值[M+H] +574，实测值574。 MS-ESI calculated [M+H] + 574, found 574.
1HNMR(400MHz,CD 3OD)δ＝8.25(dd,J＝4.5,8.4Hz,1H),8.13-8.03(m,1H),7.59(ddd,J＝1.2,7.7,14.1Hz,1H),7.45(t,J＝7.8Hz,1H),7.25(dt,J＝1.3,7.5Hz,1H),7.06(dd,J＝5.2,9.0Hz,1H),6.90(dd,J＝8.9,12.6Hz,1H),6.73-6.24(m,1H),4.47-3.90(m,4H),1.84(d,J＝13.4Hz,6H),1.51(t,J＝8.7Hz,2H). 1 HNMR (400MHz, CD 3 OD) δ = 8.25 (dd, J = 4.5, 8.4Hz, 1H), 8.13-8.03 (m, 1H), 7.59 (ddd, J = 1.2, 7.7, 14.1Hz, 1H), 7.45(t, J=7.8Hz, 1H), 7.25(dt, J=1.3, 7.5Hz, 1H), 7.06(dd, J=5.2, 9.0Hz, 1H), 6.90(dd, J=8.9, 12.6Hz ,1H),6.73-6.24(m,1H),4.47-3.90(m,4H),1.84(d,J=13.4Hz,6H),1.51(t,J=8.7Hz,2H).
实施例35Example 35
Figure PCTCN2022129958-appb-000082
Figure PCTCN2022129958-appb-000082
第一步first step
将化合物28-2(10克，45.45毫摩尔，1当量)溶于四氢呋喃(10毫升)，接着向反应液中加入二-叔-丁基二碳酸盐(19.84克，90.89毫摩尔，20.88毫升，2当量)。反应液在70摄氏度下反应12小时。反应结束后，将反应液减压浓缩，粗品经硅胶柱层析(石油醚：乙酸乙酯＝200：1-80：1)分离纯化，得到化合物35-1。Compound 28-2 (10 g, 45.45 mmol, 1 equivalent) was dissolved in tetrahydrofuran (10 ml), and then di-tert-butyl dicarbonate (19.84 g, 90.89 mmol, 20.88 ml) was added to the reaction solution , 2 equivalents). The reaction solution was reacted at 70° C. for 12 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and the crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 200:1-80:1) to obtain compound 35-1.
第二步second step
将化合物35-1(500毫克，1.56毫摩尔，1当量)，化合物B的盐酸盐(248.61毫克，1.64毫摩尔，1.05当量)，叔丁醇钠(600.34毫克，6.25毫摩尔，4当量)，三(二亚苄基丙酮)二钯(143.01毫克，156.17微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(180.73毫克，312.35微摩尔，0.2当量)溶于二氧六环(20毫升)中接着在100摄氏度氮气保护下反应12小时，反应完毕后，向反应液中加入水(50毫升)稀释，用乙酸乙酯萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥。过滤，浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-20：1)分离纯化得到化合物35-2。Compound 35-1 (500 mg, 1.56 mmol, 1 eq), compound B hydrochloride (248.61 mg, 1.64 mmol, 1.05 eq), sodium tert-butoxide (600.34 mg, 6.25 mmol, 4 eq) , tris(dibenzylideneacetone)dipalladium (143.01 mg, 156.17 micromole, 0.1 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (180.73 mg, 312.35 Micromoles, 0.2 equivalents) were dissolved in dioxane (20 milliliters) and then reacted under nitrogen protection at 100 degrees Celsius for 12 hours. After the reaction was completed, water (50 milliliters) was added to the reaction solution for dilution, and extracted with ethyl acetate ( 50 ml×2), the organic phase was washed with saturated brine (50 ml), and dried over anhydrous sodium sulfate. After filtration, the concentrated residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-20:1) to obtain compound 35-2.
MS-ESI计算值[M+H] +255，实测值255。 MS-ESI calculated [M+H] +255 , found 255.
第三步third step
将化合物35-2(104毫克，409.01微摩尔，1当量)，化合物E(155.16毫克，490.81微摩尔，1.2当量)，叔丁醇钠(117.92毫克，1.23毫摩尔，3当量)，三(二亚苄基丙酮)二钯(37.45毫克，40.90微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(47.33毫克，81.80微摩尔，0.2当量)溶于二氧六环(5毫升)中接着在100摄氏度氮气保护下反应12小时，反应完毕后，反应液过滤，浓缩的残渣经制备高效液相色谱法(色谱柱:Phenomenex Synergi C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；55％-75％，10分钟)分离得到化合物35。Compound 35-2 (104 mg, 409.01 micromoles, 1 equivalent), compound E (155.16 mg, 490.81 micromoles, 1.2 equivalents), sodium tert-butoxide (117.92 mg, 1.23 mmoles, 3 equivalents), three (di Benzylideneacetone) dipalladium (37.45 mg, 40.90 micromoles, 0.1 eq), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (47.33 mg, 81.80 micromoles, 0.2 equivalent) was dissolved in dioxane (5 milliliters) and then reacted at 100 degrees centigrade under nitrogen protection for 12 hours. After the reaction was completed, the reaction solution was filtered, and the concentrated residue was subjected to preparative high performance liquid chromatography (chromatographic column: Phenomenex Synergi C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; 55%-75%, 10 minutes) isolated to obtain compound 35.
MS-ESI计算值[M+H] +534，实测值534。 MS-ESI calculated [M+H] + 534, found 534.
1HNMR(400MHz,CD 3OD)δ＝8.29-8.23(m,1H),8.14(s,1H),7.68-7.56(m,2H),7.35-7.28(m,2H),5.93-5.87(m,1H),5.03(t,J＝6.6Hz,1H),3.87(d,J＝4.5Hz,4H),3.63(s,3H),2.66-2.58(m,2H),2.43-2.31(m,2H),1.86(s,3H),1.83(s,3H)。 1 HNMR (400MHz, CD 3 OD) δ=8.29-8.23(m,1H),8.14(s,1H),7.68-7.56(m,2H),7.35-7.28(m,2H),5.93-5.87(m ,1H),5.03(t,J=6.6Hz,1H),3.87(d,J=4.5Hz,4H),3.63(s,3H),2.66-2.58(m,2H),2.43-2.31(m, 2H), 1.86(s,3H), 1.83(s,3H).
实施例36Example 36
Figure PCTCN2022129958-appb-000083
Figure PCTCN2022129958-appb-000083
第一步first step
将化合物29-4(250毫克，500.30微摩尔，1当量)溶于二氧六环(8毫升)，接着向反应液中加入化合物B的盐酸盐(151.70毫克，1.00毫摩尔，2当量)，碳酸铯(652.03毫克，2.00毫摩尔，4当量)，4，5-双(二苯基膦)-9，9-二甲基氧杂蒽(57.90毫克，100.06微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(45.81毫克，50.03微摩尔，0.1当量)，在110摄氏度下氮气保护下反应16小时。反应完成后，反应液用硅藻土过滤，滤液减压浓缩的粗品经高效液相色谱(色谱柱：Waters Xbridge C18 15×50毫米×10微米；流动相：流动相A：碳酸氢铵水溶液；流动相B：乙腈；B％:40％-70％，10分钟)分离得到化合物36。Compound 29-4 (250 mg, 500.30 μmol, 1 eq) was dissolved in dioxane (8 ml), and then the hydrochloride of compound B (151.70 mg, 1.00 mmol, 2 eq) was added to the reaction solution , cesium carbonate (652.03 mg, 2.00 mmol, 4 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (57.90 mg, 100.06 micromol, 0.2 equiv), tris (Dibenzylideneacetone)dipalladium (45.81 mg, 50.03 micromole, 0.1 eq.) was reacted at 110°C under nitrogen protection for 16 hours. After the reaction was completed, the reaction solution was filtered with diatomaceous earth, and the crude product concentrated under reduced pressure of the filtrate was subjected to high performance liquid chromatography (chromatographic column: Waters Xbridge C18 15 × 50 millimeters × 10 microns; mobile phase: mobile phase A: ammonium bicarbonate aqueous solution; Mobile phase B: acetonitrile; B%: 40%-70%, 10 minutes) isolated to obtain compound 36.
MS-ESI计算值[M+H] +534,实测值534。 MS-ESI calculated [M+H] + 534, found 534.
1HNMR(400MHz,CD 3OD)δ＝8.36-8.28(m,1H),8.09(s,1H),7.67-7.60(m,1H),7.54(br t,J＝7.4Hz,1H),7.32-7.26(m,1H),7.24-7.13(m,1H),6.65(br dd,J＝9.1,12.4Hz,1H),5.08-5.00(m,1H),4.14-4.04(m,4H),3.62(s,3H),2.69-2.58(m,2H),2.42-2.28(m,2H),1.88(s,3H),1.84(s,3H)。 1 HNMR (400MHz, CD 3 OD) δ = 8.36-8.28 (m, 1H), 8.09 (s, 1H), 7.67-7.60 (m, 1H), 7.54 (br t, J = 7.4Hz, 1H), 7.32 -7.26(m,1H),7.24-7.13(m,1H),6.65(brdd,J＝9.1,12.4Hz,1H),5.08-5.00(m,1H),4.14-4.04(m,4H), 3.62 (s, 3H), 2.69-2.58 (m, 2H), 2.42-2.28 (m, 2H), 1.88 (s, 3H), 1.84 (s, 3H).
实施例37Example 37
Figure PCTCN2022129958-appb-000084
Figure PCTCN2022129958-appb-000084
第一步first step
将化合物34-3(250毫克，466.70微摩尔，1.0当量)，化合物A(114.56毫克，700.04微摩尔，1.5当量)，碳酸铯(456.17毫克，1.4毫摩尔，3.0当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(54.01毫克，93.34微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(85.47毫克，93.34微摩尔，0.2当量)加入二氧六环(20毫升)中，氮气置换保护，反应液加热到100摄氏下反应12小时。反应完成后，过滤，滤液经乙酸乙酯(60毫升)稀释，饱和食盐水(60毫升)洗涤。有机相经无水硫酸钠干燥，过滤，滤液经减压浓缩得到的残渣经高效液相色谱(色谱柱：Phenomenex luna C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:44％-74％，10分钟)纯化得到化合物37。Compound 34-3 (250 mg, 466.70 micromol, 1.0 equiv), compound A (114.56 mg, 700.04 micromol, 1.5 equiv), cesium carbonate (456.17 mg, 1.4 mmol, 3.0 equiv), 4,5-bis (Diphenylphosphine)-9,9-dimethylxanthene (54.01 mg, 93.34 micromol, 0.2 equiv), tris(dibenzylideneacetone)dipalladium (85.47 mg, 93.34 micromol, 0.2 equiv) Dioxane (20 ml) was added, and nitrogen replacement protection was carried out, and the reaction solution was heated to 100° C. for 12 hours. After the reaction was completed, it was filtered, and the filtrate was diluted with ethyl acetate (60 mL) and washed with saturated brine (60 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the residue obtained by concentrating the filtrate under reduced pressure was subjected to high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase Phase B: acetonitrile; B%: 44%-74%, 10 minutes) purification afforded compound 37.
MS-ESI计算值[M+H] +582,实测值582。 MS-ESI calculated [M+H] + 582, found 582.
1H NMR(400MHz,CD 3OD)δ＝8.36-8.25(m,1H),8.12-7.99(m,1H),7.65-7.55(m,1H),7.50-7.42(m,1H),7.30-7.20(m,1H),7.11-7.02(m,1H),6.86(m,1H),6.70-6.24(m,1H),4.18-4.10(m,2H),4.06(m,2H),3.87-3.77(m,1H),3.23(s,3H),2.58-2.40(m,2H),2.10-2.01(m,2H),1.88(s,3H),1.84(s,3H). 1 H NMR (400MHz, CD 3 OD) δ=8.36-8.25(m,1H),8.12-7.99(m,1H),7.65-7.55(m,1H),7.50-7.42(m,1H),7.30- 7.20(m,1H),7.11-7.02(m,1H),6.86(m,1H),6.70-6.24(m,1H),4.18-4.10(m,2H),4.06(m,2H),3.87- 3.77(m,1H),3.23(s,3H),2.58-2.40(m,2H),2.10-2.01(m,2H),1.88(s,3H),1.84(s,3H).
实施例38Example 38
Figure PCTCN2022129958-appb-000085
Figure PCTCN2022129958-appb-000085
第一步first step
将化合物34-3(250毫克，466.7微摩尔，1当量)溶于二氧六环(12毫升)中，加入化合物D(83.72毫克，700.04微摩尔，1.5当量)，碳酸铯(684.28毫克，2.1毫摩尔，4.5当量)，三(二亚苄基丙酮)二钯(85.47毫克，93.34微摩尔，0.2当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(54.01毫克，93.34微摩尔，0.2当量)，溶于二氧六环(10毫升)中接着在100摄氏度氮气保护下反应12小时，反应完毕后，向反应液中加入水(50毫升)稀释，用乙酸乙酯萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥。过滤，浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)和制备高效液相色谱法(色谱柱:Phenomenex luna C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；48％-78％，10分钟)分离得到化合物38。Compound 34-3 (250 mg, 466.7 micromoles, 1 equivalent) was dissolved in dioxane (12 ml), compound D (83.72 mg, 700.04 micromoles, 1.5 equivalents), cesium carbonate (684.28 mg, 2.1 mmoles, 4.5 equiv), tris(dibenzylideneacetone)dipalladium (85.47 mg, 93.34 micromoles, 0.2 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethyloxa Anthracene (54.01 milligrams, 93.34 micromoles, 0.2 equivalents), was dissolved in dioxane (10 milliliters) and then reacted at 100 degrees Celsius under nitrogen protection for 12 hours. After the reaction was completed, water (50 milliliters) was added to the reaction solution to dilute , extracted with ethyl acetate (50 mL×2), the organic phase was washed with saturated brine (50 mL), and dried over anhydrous sodium sulfate. Filtration, the concentrated residue was subjected to silica gel thin-layer chromatography (dichloromethane:methanol=10:1) and preparative high-performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A : aqueous formic acid; mobile phase B: acetonitrile; 48%-78%, 10 minutes) isolated to obtain compound 38.
MS-ESI计算值[M+H] +538，实测值538。 MS-ESI calculated [M+H] + 538, found 538.
1HNMR(400MHz,CD 3OD)δ＝8.29(dd,J＝4.4,8.3Hz,1H),8.05(s,1H),7.63-7.44(m,2H),7.25(br d,J＝1.2Hz,1H),7.03(dd,J＝5.1,8.9Hz,1H),6.86(dd,J＝9.0,12.7Hz,1H),6.73-6.23(m,1H),4.20(d,J＝2.8Hz,4H),1.84(d,J＝13.4Hz,6H),0.62(s,4H)。 1 HNMR (400MHz, CD 3 OD) δ = 8.29 (dd, J = 4.4, 8.3Hz, 1H), 8.05 (s, 1H), 7.63-7.44 (m, 2H), 7.25 (br d, J = 1.2Hz ,1H),7.03(dd,J=5.1,8.9Hz,1H),6.86(dd,J=9.0,12.7Hz,1H),6.73-6.23(m,1H),4.20(d,J=2.8Hz, 4H), 1.84 (d, J=13.4Hz, 6H), 0.62 (s, 4H).
实施例39Example 39
Figure PCTCN2022129958-appb-000086
Figure PCTCN2022129958-appb-000086
第一步first step
将化合物34-3(300毫克，560.03微摩尔，1.0当量)溶于二氧六环(10毫升)，氮气置换保护，加入化合物B(127.36毫克，840.05微摩尔，1.5当量)，碳酸铯(821.11毫克，2.52毫摩尔，4.5当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(48.61毫克，84.01微摩尔，0.15当量)，三(二亚苄基丙酮)二钯(76.92毫克，84.01微摩尔，0.15当量)，氮气置换保护，反应液加热到115摄氏下反应12小时。反应完成后，过滤，滤液经减压浓缩得到的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝15:1)和高效液相色谱(色谱柱：Phenomenex luna C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:39％-69％，10分钟)纯化得到化合物39。Compound 34-3 (300 mg, 560.03 micromoles, 1.0 equivalents) was dissolved in dioxane (10 ml), nitrogen displacement protection was added, compound B (127.36 mg, 840.05 micromoles, 1.5 equivalents), cesium carbonate (821.11 mg, 2.52 mmol, 4.5 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (48.61 mg, 84.01 micromol, 0.15 equiv), tris(dibenzylidene Acetone) dipalladium (76.92 mg, 84.01 micromoles, 0.15 equivalents), nitrogen replacement protection, the reaction solution was heated to 115 degrees Celsius for 12 hours. After the reaction was completed, filter, and the residue obtained by concentrating the filtrate under reduced pressure was subjected to silica gel thin-layer chromatography (dichloromethane: methanol=15:1) and high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 25 millimeters × 10 microns ; Mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 39%-69%, 10 minutes) purification to obtain compound 39.
MS-ESI计算值[M+H] +570,实测值570。 MS-ESI calculated [M+H] + 570, found 570.
1H NMR(400MHz,CD 3OD)δ＝8.34-8.25(m,1H),8.11-8.03(m,1H),7.66-7.56(m,1H),7.50-7.42(m,1H),7.29-7.22(m,1H),7.10-7.03(m,1H),6.91-6.82(m,1H),6.73-6.20(m,1H),5.00(m,1H),4.12(m,4H),2.66-2.54(m,2H),2.40-2.27(m,2H),1.88(s,3H),1.84(s,3H)。 1 H NMR (400MHz, CD 3 OD) δ=8.34-8.25(m,1H),8.11-8.03(m,1H),7.66-7.56(m,1H),7.50-7.42(m,1H),7.29- 7.22(m,1H),7.10-7.03(m,1H),6.91-6.82(m,1H),6.73-6.20(m,1H),5.00(m,1H),4.12(m,4H),2.66- 2.54(m,2H),2.40-2.27(m,2H),1.88(s,3H),1.84(s,3H).
实施例40Example 40
Figure PCTCN2022129958-appb-000087
Figure PCTCN2022129958-appb-000087
第一步first step
将化合物40-1(8克，47.29毫摩尔，1当量)溶于N，N-二甲基甲酰胺(60毫升)中，接着加入碳酸钾(19.61克，141.88毫摩尔，3当量)，在25摄氏度下搅拌0.5小时，然后加入化合物40-2(11.84克，70.94毫摩尔，1.5当量)，在60摄氏度下反应6小时，反应完毕后，向反应液中加入水(100毫升)稀释，用乙酸乙酯萃取(100毫升×2)，有机相用饱和的食盐水(100毫升)洗涤，无水硫酸钠干燥。过滤，浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-0：1)分离纯化得到化合物40-3。Compound 40-1 (8 g, 47.29 mmol, 1 eq) was dissolved in N, N-dimethylformamide (60 ml), followed by adding potassium carbonate (19.61 g, 141.88 mmol, 3 eq), in Stir at 25 degrees Celsius for 0.5 hour, then add compound 40-2 (11.84 g, 70.94 mmol, 1.5 equivalents), react at 60 degrees Celsius for 6 hours, after the reaction is complete, add water (100 ml) to the reaction solution for dilution, and use Extracted with ethyl acetate (100 mL×2), the organic phase was washed with saturated brine (100 mL), and dried over anhydrous sodium sulfate. After filtration, the concentrated residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-0:1) to obtain compound 40-3.
MS-ESI计算值[M+H] +300，实测值300。 MS-ESI calculated value [M+H] + 300, found value 300.
1HNMR(400MHz,CDCl 3)δ＝11.85(br s,1H),8.79(dd,J＝4.4,8.4Hz,1H),8.07(d,J＝2.4Hz,1H),7.59(t,J＝7.9Hz,1H),7.27-7.23(m,1H),7.20-7.12(m,1H),1.85(d,J＝13.2Hz,6H)。 1 HNMR (400MHz, CDCl 3 )δ=11.85(br s,1H),8.79(dd,J=4.4,8.4Hz,1H),8.07(d,J=2.4Hz,1H),7.59(t,J= 7.9Hz, 1H), 7.27-7.23(m, 1H), 7.20-7.12(m, 1H), 1.85(d, J=13.2Hz, 6H).
第二步second step
将化合物40-3(666毫克，2.22毫摩尔，1当量)，1-1(583.76毫克，2.89毫摩尔，1.3当量)，甲烷磺酸(213.59毫克，2.22毫摩尔，1当量)，溶于异丙醇(100毫升)中接着在90摄氏度下反应12小时，反应完毕后，将反应液冷却到室温，过滤析出的固体，得到化合物40-4。Compound 40-3 (666 mg, 2.22 mmol, 1 eq), 1-1 (583.76 mg, 2.89 mmol, 1.3 eq), methanesulfonic acid (213.59 mg, 2.22 mmol, 1 eq), were dissolved in iso Propanol (100 ml) was then reacted at 90° C. for 12 hours. After the reaction was completed, the reaction solution was cooled to room temperature, and the precipitated solid was filtered to obtain compound 40-4.
MS-ESI计算值[M +H] +467，实测值467。 MS-ESI calculated [M + H] +467 , found 467.
第三步third step
将化合物40-4(300毫克，644.81微摩尔，1当量)，化合物F(150.47毫克，967.21微摩尔，1.5当量)，碳酸铯(630.27毫克，1.93毫摩尔，3当量)，三(二亚苄基丙酮)二钯(59.05毫克，64.48微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(74.62毫克，128.96微摩尔，0.2当量)，溶于二氧六环(12毫升)中接着在115摄氏度氮气保护下反应12小时，反应完毕后，向反应液中加入水(50毫升)稀释，用乙酸乙酯萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥。过滤，浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝10：1)和高效液相色谱法(色谱柱:Waters Xbridge150×25毫米×5微米；流动相：流动相A：碳酸氢氨水溶液；流动相B：乙腈；45％-65％，9分钟)分离得到化合物40。Compound 40-4 (300 mg, 644.81 micromoles, 1 equivalent), compound F (150.47 mg, 967.21 micromoles, 1.5 equivalents), cesium carbonate (630.27 mg, 1.93 mmoles, 3 equivalents), tris(dibenzylidene acetonide) dipalladium (59.05 mg, 64.48 micromoles, 0.1 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (74.62 mg, 128.96 micromoles, 0.2 equiv) , dissolved in dioxane (12 milliliters) and then reacted at 115 degrees Celsius under nitrogen protection for 12 hours. After the reaction was completed, water (50 milliliters) was added to the reaction solution for dilution, and extracted with ethyl acetate (50 milliliters × 2) , the organic phase was washed with saturated brine (50 ml), and dried over anhydrous sodium sulfate. Filtration, concentrated residue was subjected to silica gel thin-layer chromatography plate (dichloromethane:methanol=10:1) and high performance liquid chromatography (chromatographic column: Waters Xbridge150 * 25 millimeters * 5 microns; Mobile phase: mobile phase A: bicarbonate Ammonia solution; mobile phase B: acetonitrile; 45%-65%, 9 minutes) isolated compound 40.
MS-ESI计算值[M+H] +504，实测值504。 MS-ESI calculated [M+H] + 504, found 504.
1HNMR(400MHz,CD 3OD)δ＝8.52(dd,J＝4.5,8.2Hz,1H),8.01(d,J＝3.5Hz,1H),7.64-7.47(m,3H),7.29-7.21(m,1H),6.91(t,J＝8.1Hz,1H),6.32(dd,J＝1.3,8.1Hz,1H),4.11(d,J＝7.9Hz,2H),3.97 (br d,J＝7.9Hz,2H),3.69(s,3H),1.86(d,J＝13.6Hz,6H),1.54(t,J＝8.6Hz,2H)。 1 HNMR (400MHz, CD 3 OD) δ = 8.52 (dd, J = 4.5, 8.2Hz, 1H), 8.01 (d, J = 3.5Hz, 1H), 7.64-7.47 (m, 3H), 7.29-7.21 ( m,1H),6.91(t,J=8.1Hz,1H),6.32(dd,J=1.3,8.1Hz,1H),4.11(d,J=7.9Hz,2H),3.97 (br d,J= 7.9Hz, 2H), 3.69(s, 3H), 1.86(d, J=13.6Hz, 6H), 1.54(t, J=8.6Hz, 2H).
实施例41Example 41
Figure PCTCN2022129958-appb-000088
Figure PCTCN2022129958-appb-000088
第一步first step
将化合物28-3(88.75毫克，177.61微摩尔，1当量)，化合物L(70毫克，230.89微摩尔，1.3当量)，碳酸铯(173.61毫克，532.83微摩尔，3当量)，醋酸钯(3.99毫克，17.76微摩尔，0.1当量)，正丁基-二(1-金刚烷基)膦(12.74毫克，35.52微摩尔，0.2当量)，溶于甲苯(10毫升)和水(1毫升)中接着在120摄氏度氮气保护下反应10小时，反应完毕后，反应液过滤，浓缩的残渣经高效液相色谱法(色谱柱:Phenomenex luna C18 150×40毫米×15微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；60％-90％，10分钟)分离得到化合物41-1。Compound 28-3 (88.75 mg, 177.61 micromoles, 1 equivalent), compound L (70 mg, 230.89 micromoles, 1.3 equivalents), cesium carbonate (173.61 mg, 532.83 micromoles, 3 equivalents), palladium acetate (3.99 mg , 17.76 micromoles, 0.1 equivalents), n-butyl-bis(1-adamantyl)phosphine (12.74 mg, 35.52 micromoles, 0.2 equivalents), dissolved in toluene (10 milliliters) and water (1 milliliters) followed by 120 degrees centigrade reacted under nitrogen protection for 10 hours, after the reaction was completed, the reaction solution was filtered, and the concentrated residue was subjected to high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 40 mm × 15 microns; mobile phase: mobile phase A: formic acid aqueous solution ; mobile phase B: acetonitrile; 60%-90%, 10 minutes) isolated to obtain compound 41-1.
MS-ESI计算值[M+H] +616，实测值616。 MS-ESI calculated [M+H] + 616, found 616.
第二步second step
将化合物41-1(60毫克，95.25微摩尔，1当量)溶于乙酸乙酯(2毫升)，接着向反应液中加入浓硫酸(45.71毫克，476.25微摩尔，25.39微升，5当量)，在20摄氏度下反应0.5小时。反应完成后，浓缩得到粗品化合物41-2(硫酸盐)。Compound 41-1 (60 mg, 95.25 micromoles, 1 equivalent) was dissolved in ethyl acetate (2 ml), then concentrated sulfuric acid (45.71 mg, 476.25 micromoles, 25.39 microliters, 5 equivalents) was added to the reaction solution, React at 20°C for 0.5 hours. After the reaction was completed, it was concentrated to obtain crude compound 41-2 (sulfate).
MS-ESI计算值[M+H] +516,实测值516。 MS-ESI calculated [M+H] + 516, found 516.
第三步third step
将化合物41-2(60毫克，97.72微摩尔，1当量，硫酸盐)溶于甲醇(2毫升)，接着向反应液中加入甲醛(14.67毫克，488.58微摩尔，13.46微升，5当量)，氰基硼氢化钠(12.28毫克，195.43微摩尔，2当量)和N，N-二异丙基乙胺(37.89毫克，293.15微摩尔，51.06微升，3当量)，在20摄氏度下反应1小时。反应完成后，反应液过滤浓缩的残渣经高效液相色谱(色谱柱：Phenomenex luna C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：18％-38％，10分钟)分离得到41(甲酸盐)。Compound 41-2 (60 mg, 97.72 micromoles, 1 equiv, sulfate) was dissolved in methanol (2 ml), then formaldehyde (14.67 mg, 488.58 micromoles, 13.46 microliters, 5 equivs) was added to the reaction solution, Sodium cyanoborohydride (12.28 mg, 195.43 micromol, 2 equiv) and N,N-diisopropylethylamine (37.89 mg, 293.15 micromol, 51.06 microliter, 3 equiv) were reacted at 20 °C for 1 hour . After the reaction was completed, the concentrated residue of the reaction solution was filtered through high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 25 millimeters × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 18 %-38%, 10 min) to isolate 41 (formate salt).
MS-ESI计算值[M+H] +530,实测值530。 MS-ESI calculated value [M+H] + 530, found value 530.
1HNMR(400MHz,CD 3OD)δ＝8.51(s,1H),8.27-8.22(m,1H),8.17(s,1H),7.82-7.75(m,1H),7.72-7.58(m,2H),7.38-7.30(m,1H),6.69-6.61(m,1H),4.30(br s,2H),4.06(s,2H),3.70(s,4H),2.88(s,3H),2.71(br t,J＝10.4Hz,2H),2.45-2.33(m,2H),1.85(d,J＝13.5Hz,6H)。 1 HNMR (400MHz, CD 3 OD) δ=8.51(s,1H),8.27-8.22(m,1H),8.17(s,1H),7.82-7.75(m,1H),7.72-7.58(m,2H ),7.38-7.30(m,1H),6.69-6.61(m,1H),4.30(br s,2H),4.06(s,2H),3.70(s,4H),2.88(s,3H),2.71 (br t, J=10.4Hz, 2H), 2.45-2.33(m, 2H), 1.85(d, J=13.5Hz, 6H).
实施例42Example 42
Figure PCTCN2022129958-appb-000089
Figure PCTCN2022129958-appb-000089
第一步first step
将化合物29-3(500毫克，2.27毫摩尔1当量)，化合物F(530.27毫克，3.41毫摩尔，1.5当量)，叔丁醇钠(873.51毫克，9.09毫摩尔，3当量)，三(二亚苄基丙酮)二钯(208.08毫克，227.23微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(262.96毫克，454.47微摩尔，0.2当量)溶于二氧六环(15毫升)中接着在100摄氏度氮气保护下反应12小时，反应完毕后，向反应液中加入水(50毫升)稀释，用乙酸乙酯萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥。过滤，浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-0：1)分离纯化得到化合物42-1。Compound 29-3 (500 mg, 2.27 mmol 1 equivalent), compound F (530.27 mg, 3.41 mmol, 1.5 equivalents), sodium tert-butoxide (873.51 mg, 9.09 mmol, 3 equivalents), three (di Benzylacetone) dipalladium (208.08 mg, 227.23 micromoles, 0.1 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (262.96 mg, 454.47 micromoles, 0.2 equiv ) was dissolved in dioxane (15 milliliters) and then reacted at 100 degrees Celsius under nitrogen protection for 12 hours. After the reaction was completed, water (50 milliliters) was added to the reaction solution for dilution, and extracted with ethyl acetate (50 milliliters × 2) , the organic phase was washed with saturated brine (50 ml), and dried over anhydrous sodium sulfate. After filtration, the concentrated residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-0:1) to obtain compound 42-1.
MS-ESI计算值[M+H] +259，实测值259。 MS-ESI calculated [M+H] +259 , found 259.
1HNMR(400MHz,CDCl 3)δ＝6.60(dd,J＝8.7,12.5Hz,1H),6.17-6.05(m,1H),4.36(dd,J＝2.4,8.4Hz,2H),4.25-4.18(m,2H),3.65(s,3H),1.43(t,J＝8.4Hz,2H)。 1 HNMR (400MHz, CDCl 3 )δ=6.60(dd, J=8.7,12.5Hz,1H),6.17-6.05(m,1H),4.36(dd,J=2.4,8.4Hz,2H),4.25-4.18 (m, 2H), 3.65 (s, 3H), 1.43 (t, J=8.4Hz, 2H).
第二步second step
将化合物42-1(250毫克，968.1微摩尔，1当量)，化合物40-3(377.14毫克，1.26毫摩尔，1.3当量)，碳酸铯(946.27毫克，2.9毫摩尔，3当量)，三(二亚苄基丙酮)二钯(88.65毫克，96.81微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(112.03毫克，193.62微摩尔，0.2当量)溶于二氧六环(12毫升)中接着在110摄氏度氮气保护下反应12小时，反应完毕后，向反应液中加入水(50毫升)稀释，用二氯甲烷萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥。过滤，浓缩的残渣经制备高效液相色谱法(色谱柱:Phenomenex luna C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；38％-68％，10分钟)分离得到化合物42。Compound 42-1 (250 mg, 968.1 micromole, 1 equivalent), compound 40-3 (377.14 mg, 1.26 mmol, 1.3 equivalent), cesium carbonate (946.27 mg, 2.9 mmol, 3 equivalents), three (two Benzylideneacetone) dipalladium (88.65 mg, 96.81 micromoles, 0.1 eq), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (112.03 mg, 193.62 micromoles, 0.2 equivalent) was dissolved in dioxane (12 milliliters) and then reacted at 110 degrees Celsius under nitrogen protection for 12 hours. After the reaction was completed, water (50 milliliters) was added to the reaction solution for dilution, and extracted with dichloromethane (50 milliliters × 2 ), the organic phase was washed with saturated brine (50 ml), and dried over anhydrous sodium sulfate. Filtration, the concentrated residue was subjected to preparative high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 25 millimeters × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; 38%-68%, 10 minutes) to isolate compound 42.
MS-ESI计算值[M+H] +522，实测值522。 MS-ESI calculated [M+H] + 522, found 522.
1HNMR(400MHz,CD 3OD)δ＝8.49(dd,J＝4.5,8.4Hz,1H),7.98(d,J＝3.2Hz,1H),7.70-7.48(m,2H),7.37-7.18(m,2H),6.72(dd,J＝9.1,12.5Hz,1H),4.29(dd,J＝2.0,8.3Hz,2H),4.24-4.17(m,2H),3.64(s,3H),1.86(d,J＝13.6Hz,6H),1.51(t,J＝8.6Hz,2H)。 1 HNMR (400MHz, CD 3 OD) δ = 8.49 (dd, J = 4.5, 8.4Hz, 1H), 7.98 (d, J = 3.2Hz, 1H), 7.70-7.48 (m, 2H), 7.37-7.18 ( m,2H),6.72(dd,J=9.1,12.5Hz,1H),4.29(dd,J=2.0,8.3Hz,2H),4.24-4.17(m,2H),3.64(s,3H),1.86 (d, J=13.6Hz, 6H), 1.51 (t, J=8.6Hz, 2H).
实施例43Example 43
Figure PCTCN2022129958-appb-000090
Figure PCTCN2022129958-appb-000090
第一步first step
将化合物35-1(550毫克，1.72毫摩尔，1当量)溶于二氧六环(20毫升)，接着向反应液中加入化合物F(280.63毫克，1.80毫摩尔，1.05当量)，碳酸铯(2.24克，6.87毫摩尔，4当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(198.80毫克，343.58微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(157.31毫克，171.79微摩尔，0.1当量)，置换三次氮气，并在氮气氛围下在100摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1到25：1)纯化得到化合物43-1。Compound 35-1 (550 mg, 1.72 mmol, 1 eq) was dissolved in dioxane (20 ml), then compound F (280.63 mg, 1.80 mmol, 1.05 eq), cesium carbonate ( 2.24 g, 6.87 mmol, 4 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (198.80 mg, 343.58 micromol, 0.2 equiv), tris(diphenylene Acetone) dipalladium (157.31 mg, 171.79 micromole, 0.1 equiv), three nitrogen replacements, and reacted at 100 degrees Celsius under nitrogen atmosphere for 12 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1 to 25:1) to obtain compound 43-1.
MS-ESI计算值[M+H] +359,实测值359。 MS-ESI calculated [M+H] + 359, found 359.
第二步second step
将化合物43-1(360毫克，1.00毫摩尔，1当量)溶于乙酸乙酯(4毫升)，接着向反应液中加入浓硫酸(502.70毫克，5.02毫摩尔，273.21微升，纯度：98％，5当量)，在25摄氏度下反应0.5小时。反应完成后，用碳酸氢钠的饱和溶液将反应液的pH调至8，滤液浓缩，得到粗品化合物43-2。Compound 43-1 (360 mg, 1.00 mmol, 1 eq) was dissolved in ethyl acetate (4 ml), then concentrated sulfuric acid (502.70 mg, 5.02 mmol, 273.21 μl, purity: 98% , 5 equivalents), reacted at 25 degrees Celsius for 0.5 hours. After the reaction was completed, the pH of the reaction solution was adjusted to 8 with a saturated solution of sodium bicarbonate, and the filtrate was concentrated to obtain the crude compound 43-2.
MS-ESI计算值[M+H] +259,实测值259。 MS-ESI calculated [M+H] +259 , found 259.
第三步third step
将化合物43-2(80毫克，309.79微摩尔，1当量)溶于二氧六环(5毫升)，然后向反应液中加入化合物40-3(111.40毫克，371.75毫微摩尔，1.2当量)，叔丁醇钠(89.32毫克，929.37微摩尔，3当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(35.85毫克，91.96微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(28.37毫克，30.98微摩尔，0.1当量)，置换三次氮气，并在氮气氛围下在115摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝50：1到1：2，然后二氯甲烷：甲醇＝100：1到50：1)纯化得到的粗品经高效液相色谱(色谱柱：Phenomenex Luna C18 150×40毫米×15微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％：47％-77％，10分钟)分离得到化合物43。Compound 43-2 (80 mg, 309.79 micromoles, 1 equiv) was dissolved in dioxane (5 ml), and compound 40-3 (111.40 mg, 371.75 nanomoles, 1.2 equivs) was added to the reaction solution, Sodium tert-butoxide (89.32 mg, 929.37 micromoles, 3 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (35.85 mg, 91.96 micromoles, 0.2 equiv), Tris(dibenzylideneacetone)dipalladium (28.37 mg, 30.98 micromole, 0.1 equiv), nitrogen was replaced three times, and reacted at 115° C. for 12 hours under nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=50:1 to 1:2, then dichloromethane:methanol=100:1 to 50:1) and the crude product obtained was purified by high performance liquid chromatography (column: Phenomenex Luna C18 150 × 40 mm × 15 microns; mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 47%-77%, 10 minutes) to obtain compound 43.
MS-ESI计算值[M+H] +522,实测值522。 MS-ESI calculated value [M+H] + 522, found value 522.
1H NMR(400MHz,CD 3OD)δ＝8.48(dd,J＝4.3,8.2Hz,1H),8.08(d,J＝3.5Hz,1H),7.72-7.56(m,2H),7.47(dd,J＝2.9,11.3Hz,1H),7.31(dt,J＝1.2,7.5Hz,1H),6.04(dd,J＝2.9,10.3Hz,1H),4.14(d,J＝7.9Hz,2H),4.01(br d,J＝8.1Hz,2H),3.68(s,3H),1.89(d,J＝13.5Hz,6H),1.57(t,J＝8.6Hz,2H)。 1 H NMR (400MHz, CD 3 OD) δ = 8.48 (dd, J = 4.3, 8.2Hz, 1H), 8.08 (d, J = 3.5Hz, 1H), 7.72-7.56 (m, 2H), 7.47 (dd ,J=2.9,11.3Hz,1H),7.31(dt,J=1.2,7.5Hz,1H),6.04(dd,J=2.9,10.3Hz,1H),4.14(d,J=7.9Hz,2H) , 4.01 (br d, J = 8.1Hz, 2H), 3.68 (s, 3H), 1.89 (d, J = 13.5Hz, 6H), 1.57 (t, J = 8.6Hz, 2H).
实施例44Example 44
Figure PCTCN2022129958-appb-000091
Figure PCTCN2022129958-appb-000091
第一步first step
将化合物35-1(500毫克，1.56毫摩尔，1当量)溶于二氧六环(15毫升)，接着向反应液中加入化合物D(196.11毫克，1.64毫摩尔，1.5当量)，碳酸铯(2.04克，6.25毫摩尔，4当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(180.73毫克，312.35微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(143.01毫克，156.17微摩尔，0.1当量)，置换三次氮气，并在氮气氛围下在100摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1到25：1)纯化得到化合物44-1。Compound 35-1 (500 mg, 1.56 mmol, 1 eq) was dissolved in dioxane (15 ml), then compound D (196.11 mg, 1.64 mmol, 1.5 eq), cesium carbonate ( 2.04 g, 6.25 mmol, 4 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (180.73 mg, 312.35 micromol, 0.2 equiv), tris(diphenylene Acetone) dipalladium (143.01 mg, 156.17 micromole, 0.1 eq.), three nitrogen replacements, and reacted at 100 degrees Celsius for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1 to 25:1) to obtain compound 44-1.
MS-ESI计算值[M+H] +323,实测值323。 MS-ESI calculated [M+H] + 323, found 323.
第二步second step
将化合物44-1(360毫克，1.12毫摩尔，1当量)溶于乙酸乙酯(7毫升)，接着向反应液中加入浓硫酸(558.81毫克，5.58毫摩尔，303.70微升，纯度：98％，5当量)，在25摄氏度下反应0.5小时。反应完成后，用碳酸氢钠的饱和溶液将反应液的pH调至8，滤液浓缩，得到粗品化合物44-2(240毫克，黄色油，收率：96.70％)。Compound 44-1 (360 mg, 1.12 mmol, 1 eq) was dissolved in ethyl acetate (7 ml), then concentrated sulfuric acid (558.81 mg, 5.58 mmol, 303.70 μl, purity: 98% , 5 equivalents), reacted at 25 degrees Celsius for 0.5 hours. After the reaction was completed, the pH of the reaction solution was adjusted to 8 with a saturated solution of sodium bicarbonate, and the filtrate was concentrated to obtain crude compound 44-2 (240 mg, yellow oil, yield: 96.70%).
MS-ESI计算值[M+H] +223,实测值223。 MS-ESI calculated value [M+H] + 223, found value 223.
第三步third step
将化合物44-2(240毫克，1.08微摩尔，1当量)溶于二氧六环(15毫升)，接着向反应液中加入化合物40-3(388.31毫克，1.30毫微摩尔，1.2当量)，碳酸铯(1.06毫克，3.24毫摩尔，3当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(124.96毫克，215.97微摩尔，0.2当量)，三(二亚苄基丙酮)二钯(98.88毫克，107.98微摩尔，0.1当量)，置换三次氮气，并在氮气氛围下在115摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝50：1到1：2，然后二氯甲烷：甲醇＝100：1到50：1)纯化得到的粗品经高效液相色谱(色谱柱：Phenomenex Luna C18 150×25毫米×10微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％:51％-81％，10分钟)分离得到化合物44。Compound 44-2 (240 mg, 1.08 micromol, 1 equiv) was dissolved in dioxane (15 ml), and then compound 40-3 (388.31 mg, 1.30 nanomol, 1.2 equiv) was added to the reaction solution, Cesium carbonate (1.06 mg, 3.24 mmol, 3 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (124.96 mg, 215.97 micromol, 0.2 equiv), tris( Dibenzylideneacetone) dipalladium (98.88 mg, 107.98 micromole, 0.1 equiv), nitrogen was replaced three times, and reacted at 115 degrees Celsius under nitrogen atmosphere for 12 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=50:1 to 1:2, then dichloromethane:methanol=100:1 to 50:1) and the crude product obtained was purified by high performance liquid chromatography (column: Phenomenex Luna C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 51%-81%, 10 minutes) to isolate compound 44.
MS-ESI计算值[M+H] +486,实测值486. MS-ESI calculated value [M+H] + 486, measured value 486.
1H NMR(400MHz,CD 3OD)δ＝8.50(dd,J＝4.3,8.1Hz,1H),8.07(d,J＝3.3Hz,1H),7.71-7.58(m,2H),7.43(dd,J＝2.9,11.3Hz,1H),7.34-7.24(m,1H),5.96(dd,J＝3.0,10.5Hz,1H),4.02(s,4H),3.67(s,3H),1.89(d,J＝13.5Hz,6H),0.69(s,4H)。 1 H NMR (400MHz, CD 3 OD) δ=8.50(dd, J=4.3, 8.1Hz, 1H), 8.07(d, J=3.3Hz, 1H), 7.71-7.58(m, 2H), 7.43(dd ,J=2.9,11.3Hz,1H),7.34-7.24(m,1H),5.96(dd,J=3.0,10.5Hz,1H),4.02(s,4H),3.67(s,3H),1.89( d, J=13.5Hz, 6H), 0.69(s, 4H).
实施例45Example 45
Figure PCTCN2022129958-appb-000092
Figure PCTCN2022129958-appb-000092
第一步first step
将化合物29-3(1.8克，8.18毫摩尔，1当量)溶于四氢呋喃(25毫升)中，加入二-叔-丁基二碳酸酐(2.68克，12.27毫摩尔，1.5当量)，接着在70摄氏度氮气保护下反应12小时。反应完毕后，将反应液减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-10：1)分离纯化得到化合物45-1。Compound 29-3 (1.8 g, 8.18 mmol, 1 eq) was dissolved in tetrahydrofuran (25 ml), and di-tert-butyl dicarbonic anhydride (2.68 g, 12.27 mmol, 1.5 eq) was added, followed by 70 The reaction was carried out for 12 hours at 10°C under the protection of nitrogen. After the reaction was completed, the residue obtained by concentrating the reaction solution under reduced pressure was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-10:1) to obtain compound 45-1.
MS-ESI计算值[M+H] +367，实测值367。 MS-ESI calculated [M+H] +367 , found 367.
1HNMR(400MHz,CDCl 3)δ＝8.11-7.96(m,1H),6.91(t,J＝8.6Hz,2H),3.89(s,3H),1.54(s,10H)。 1 H NMR (400 MHz, CDCl 3 ) δ=8.11-7.96 (m, 1H), 6.91 (t, J=8.6 Hz, 2H), 3.89 (s, 3H), 1.54 (s, 10H).
第二步second step
将化合物45-1(1.5克，4.69毫摩尔，1当量)，化合物A(805.05毫克，4.92毫摩尔，1.05当量)，碳酸铯(6.11克，18.74毫摩尔，4当量)，三(二亚苄基丙酮)二钯(429.04毫克，468.52微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(542.19毫克，937.05微摩尔，0.2当量)溶于二氧六环(20毫升)中接着在100摄氏度氮气保护下反应12小时，反应完毕后，向反应液中加入水(50毫升)稀释，用乙酸乙酯萃取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥。过滤，浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-20：1)分离纯化得到化合物45-2。Compound 45-1 (1.5 g, 4.69 mmol, 1 eq), compound A (805.05 mg, 4.92 mmol, 1.05 eq), cesium carbonate (6.11 g, 18.74 mmol, 4 eq), tris(dibenzylidene Dipalladium (429.04 mg, 468.52 micromoles, 0.1 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (542.19 mg, 937.05 micromoles, 0.2 equiv) Dissolve in dioxane (20 milliliters) and then react at 100 degrees Celsius under nitrogen protection for 12 hours. After the reaction is completed, add water (50 milliliters) to the reaction solution for dilution, and extract with ethyl acetate (50 milliliters × 2), The organic phase was washed with saturated brine (50 mL), and dried over anhydrous sodium sulfate. After filtration, the concentrated residue was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-20:1) to obtain compound 45-2.
MS-ESI计算值[M+H] +367，实测值367。 MS-ESI calculated [M+H] +367 , found 367.
第三步third step
将化合物45-2(900毫克，2.46毫摩尔，1当量)溶于乙酸乙酯(10毫升)中，再加入硫酸(1.28克，654.61毫摩尔，5当量)，接着在25摄氏度下反应0.5小时，反应完毕后，向反应液中加入饱和的碳酸氢钠溶液(50毫升)，用乙酸乙酯萃取(50毫升×3)，有机相用饱和的食盐水(40毫升×2)洗涤，无水硫酸钠干燥。过滤，浓缩得到化合物45-3。Compound 45-2 (900 mg, 2.46 mmol, 1 equivalent) was dissolved in ethyl acetate (10 ml), and sulfuric acid (1.28 g, 654.61 mmol, 5 equivalents) was added, followed by reaction at 25 degrees Celsius for 0.5 hours , after the reaction was completed, a saturated sodium bicarbonate solution (50 ml) was added to the reaction solution, extracted with ethyl acetate (50 ml × 3), the organic phase was washed with saturated brine (40 ml × 2), anhydrous Na2SO4 dried. Filtration and concentration afforded compound 45-3.
MS-ESI计算值[M+H] +267，实测值267。 MS-ESI calculated [M+H] +267 , found 267.
第四步the fourth step
将化合物45-3(100毫克，375.50微摩尔，1当量)，化合物40-3(112.53毫克，375.50毫摩尔，1当量)，叔丁醇钠(108.26毫克，1.13毫摩尔，3当量)，三(二亚苄基丙酮)二钯(34.39毫克，37.55微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(43.45毫克，75.10微摩尔，0.2当量)溶于二氧六环(6毫升)中接着在110摄氏度氮气保护下反应12小时，反应完毕后，向反应液中加入水(50毫升)稀释，用二氯甲烷取(50毫升×2)，有机相用饱和的食盐水(50毫升)洗涤，无水硫酸钠干燥。 过滤，浓缩的残渣经制备高效液相色谱法(色谱柱:Phenomenex luna C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；38％-68％，10分钟)分离得到化合物45。Compound 45-3 (100 mg, 375.50 micromol, 1 equivalent), compound 40-3 (112.53 mg, 375.50 mmol, 1 equivalent), sodium tert-butoxide (108.26 mg, 1.13 mmol, 3 equivalents), three (dibenzylideneacetone)dipalladium (34.39 mg, 37.55 micromoles, 0.1 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (43.45 mg, 75.10 micromoles , 0.2 equivalents) were dissolved in dioxane (6 milliliters) and then reacted at 110 degrees centigrade under nitrogen protection for 12 hours. After the reaction was completed, water (50 milliliters) was added to the reaction solution for dilution, and dichloromethane was used to extract (50 milliliters) ×2), the organic phase was washed with saturated brine (50 ml), and dried over anhydrous sodium sulfate. Filtration, the concentrated residue was subjected to preparative high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 25 millimeters × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; 38%-68%, 10 minutes) to isolate compound 45.
MS-ESI计算值[M+H] +530，实测值530。 MS-ESI calculated [M+H] + 530, found 530.
1HNMR(400MHz,CD 3OD)δ＝8.48(dd,J＝4.3,8.1Hz,1H),8.06(d,J＝3.5Hz,1H),7.76-7.54(m,2H),7.42(dd,J＝3.0,11.3Hz,1H),7.34-7.25(m,1H),5.91(dd,J＝3.0,10.5Hz,1H),3.91-3.81(m,5H),3.64(s,3H),3.25(s,3H),2.64-2.44(m,2H),2.16-2.03(m,2H),1.88(d,J＝13.6Hz,6H)。 1 HNMR (400MHz, CD 3 OD) δ = 8.48 (dd, J = 4.3, 8.1Hz, 1H), 8.06 (d, J = 3.5Hz, 1H), 7.76-7.54 (m, 2H), 7.42 (dd, J=3.0,11.3Hz,1H),7.34-7.25(m,1H),5.91(dd,J=3.0,10.5Hz,1H),3.91-3.81(m,5H),3.64(s,3H),3.25 (s, 3H), 2.64-2.44 (m, 2H), 2.16-2.03 (m, 2H), 1.88 (d, J=13.6Hz, 6H).
实施例46Example 46
Figure PCTCN2022129958-appb-000093
Figure PCTCN2022129958-appb-000093
第一步first step
将化合物35-2(75毫克，294.96微摩尔，1当量)，化合物40-3(106.07毫克，353.95微摩尔，1.2当量)，叔丁醇钠(85.04毫克，884.87微摩尔，3当量)，三(二亚苄基丙酮)二钯(27.01毫克，29.50微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(34.13毫克，58.99微摩尔，0.2当量)，溶于二氧六环(5毫升)中接着在100摄氏度氮气保护下反应12小时，反应完毕后，用乙酸乙酯萃取(5毫升×2)，有机相用饱和的食盐水(5毫升×2)洗涤，无水硫酸钠干燥。过滤，浓缩的残渣经制备高效液相色谱法(色谱柱:Phenomenex Synergi C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液-流动相B：乙腈；51％-71％，10分钟)分离得到化合物46。Compound 35-2 (75 mg, 294.96 micromoles, 1 equivalent), compound 40-3 (106.07 mg, 353.95 micromoles, 1.2 equivalents), sodium tert-butoxide (85.04 mg, 884.87 micromoles, 3 equivalents), three (dibenzylideneacetone)dipalladium (27.01mg, 29.50μmol, 0.1eq), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (34.13mg, 58.99μmol , 0.2 equivalents), dissolved in dioxane (5 milliliters) and then reacted at 100 degrees Celsius under nitrogen protection for 12 hours. After the reaction was completed, extracted with ethyl acetate (5 milliliters × 2), and the organic phase was saturated with brine (5 mL×2) and dried over anhydrous sodium sulfate. Filtration, the concentrated residue was subjected to preparative high-performance liquid chromatography (chromatographic column: Phenomenex Synergi C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution-mobile phase B: acetonitrile; 51%-71%, 10 minutes) to isolate compound 46.
MS-ESI计算值[M+H] +518，实测值518。 MS-ESI calculated [M+H] + 518, found 518.
1HNMR(400MHz,CD 3OD)δ＝8.50-8.41(m,1H),8.05(d,J＝3.5Hz,1H),7.67-7.55(m,2H),7.45-7.37(m,1H),7.33-7.22(m,1H),5.94-5.84(m,1H),5.08-4.89(m,1H),3.88(d,J＝4.5Hz,4H),3.63(s,3H),2.67-2.56(m,2H),2.46-2.32(m,2H),1.86(d,J＝13.4Hz,6H)。 1 HNMR (400MHz, CD 3 OD) δ = 8.50-8.41 (m, 1H), 8.05 (d, J = 3.5Hz, 1H), 7.67-7.55 (m, 2H), 7.45-7.37 (m, 1H), 7.33-7.22(m,1H),5.94-5.84(m,1H),5.08-4.89(m,1H),3.88(d,J=4.5Hz,4H),3.63(s,3H),2.67-2.56( m, 2H), 2.46-2.32 (m, 2H), 1.86 (d, J=13.4Hz, 6H).
实施例47Example 47
Figure PCTCN2022129958-appb-000094
Figure PCTCN2022129958-appb-000094
第一步first step
将化合物28-2(6克，27.3毫摩尔，1当量)溶于四氢呋喃(80毫升)，接着向反应液中加入三乙胺(9.66克，95.4毫摩尔，3.5当量)，三氟乙酸酐(17.2克，81.8毫摩尔，3当量)，在0摄氏度下反应1小时。反应完成后，加碳酸氢钠饱和水溶液(250毫升)，水(20毫升)，乙酸乙酯萃取(100毫升×3)，合并的有机相用饱和食盐水溶液(100毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝200：1-50：1)纯化得到化合物47-1。Compound 28-2 (6 g, 27.3 mmol, 1 eq) was dissolved in tetrahydrofuran (80 ml), then triethylamine (9.66 g, 95.4 mmol, 3.5 eq), trifluoroacetic anhydride ( 17.2 g, 81.8 mmol, 3 equivalents), reacted at 0 degrees Celsius for 1 hour. After completion of the reaction, add saturated aqueous sodium bicarbonate (250 ml), water (20 ml), and extract with ethyl acetate (100 ml × 3), and the combined organic phases are washed with saturated aqueous salt solution (100 ml × 2), and the organic phase After drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 200:1-50:1) to obtain compound 47-1.
1HNMR(400MHz,CDCl 3)δ＝8.54(br s,1H),8.12(dd,J＝2.9,9.5Hz,1H),7.15(dd,J＝2.9,7.6Hz,1H),3.92(s,3H)。 1 HNMR (400MHz, CDCl 3 )δ=8.54(br s,1H),8.12(dd,J=2.9,9.5Hz,1H),7.15(dd,J=2.9,7.6Hz,1H),3.92(s, 3H).
第二步second step
将化合物47-1(8.1克，25.6毫摩尔，1当量)溶于四氢呋喃(80毫升)，接着在-78摄氏度下向反应液中加入正丁基锂(2.5摩尔每升，25.6毫升，2.5当量)，在-78摄氏度下反应1小时。然后接着在-78摄氏度下向反应液中加入化合物M(5.41克，25.6毫摩尔，1当量)，在-78摄氏度下反应1小时。反应完成后，加饱和氯化铵水溶液(200毫升)稀释，乙酸乙酯萃取(60毫升×3)，合并的有机相用饱和食盐水溶液(100毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1-4：1)纯化的残渣经高效液相色谱(色谱柱：Welch Ultimate XB-CN 250×70×10微米；流动相：流动相A：正己烷；流动相B：乙醇；B％:1％-45％，15分钟)分离得到化合物47-2。Compound 47-1 (8.1 g, 25.6 mmol, 1 eq) was dissolved in tetrahydrofuran (80 ml), and then n-butyl lithium (2.5 moles per liter, 25.6 ml, 2.5 eq) was added to the reaction solution at -78°C ), react at -78°C for 1 hour. Then compound M (5.41 g, 25.6 mmol, 1 equivalent) was added to the reaction solution at -78°C, and reacted at -78°C for 1 hour. After completion of the reaction, dilute with saturated aqueous ammonium chloride (200 ml), extract with ethyl acetate (60 ml × 3), wash the combined organic phase with saturated saline solution (100 ml × 2), and wash the organic phase with anhydrous sodium sulfate Dry, filter and concentrate. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-4:1), and the residue was purified by high performance liquid chromatography (column: Welch Ultimate XB-CN 250×70×10 microns; mobile phase: mobile phase Phase A: n-hexane; mobile phase B: ethanol; B%: 1%-45%, 15 minutes) isolated to obtain compound 47-2.
MS-ESI计算值[M+H] +449,实测值349，393。 MS-ESI calculated value [M+H] + 449, found value 349,393.
1HNMR(400MHz,CD 3OD)δ＝7.46(dd,J＝3.1,9.1Hz,1H),7.00(dd,J＝3.1,9.2Hz,1H),4.21-4.05(m,3H),3.81(s,4H),2.84-2.74(m,2H),2.68-2.53(m,2H),1.45(s,9H)。 1 HNMR (400MHz, CD 3 OD) δ = 7.46 (dd, J = 3.1, 9.1 Hz, 1H), 7.00 (dd, J = 3.1, 9.2 Hz, 1H), 4.21-4.05 (m, 3H), 3.81 ( s,4H), 2.84-2.74(m,2H), 2.68-2.53(m,2H), 1.45(s,9H).
第三步third step
将化合物47-2(390毫克，870微摩尔，1当量)溶于二氯甲烷(4毫升)，接着向反应液中加入三氟乙酸(616毫克，5.40毫摩尔，6.21当量)，在20摄氏度下反应2小时。反应完成后，浓缩得到化合物47-3(三 氟乙酸盐)。Compound 47-2 (390 mg, 870 micromole, 1 eq) was dissolved in dichloromethane (4 ml), then trifluoroacetic acid (616 mg, 5.40 mmol, 6.21 eq) was added to the reaction solution, and the The reaction was carried out for 2 hours. After the reaction was completed, it was concentrated to obtain compound 47-3 (trifluoroacetate salt).
MS-ESI计算值[M+H] +349,实测值349。 MS-ESI calculated [M+H] + 349, found 349.
第四步the fourth step
将化合物47-3(300毫克，649微摩尔，1当量，三氟乙酸盐)溶于甲醇(3毫升)，接着向反应液中加入甲醛水溶液(263毫克，3.24毫摩尔，纯度：37％，5当量)，氰基硼氢化钠(102毫克，1.62毫摩尔，2.5当量)，在20摄氏度下反应1小时。反应完成后，浓缩的残渣经高效液相色谱(色谱柱：Phenomenex Synergi C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：0％-30％，10分钟)分离得到化合物47-4。Compound 47-3 (300 mg, 649 micromole, 1 equiv, trifluoroacetate salt) was dissolved in methanol (3 mL), and aqueous formaldehyde (263 mg, 3.24 mmol, purity: 37% , 5 equivalents), sodium cyanoborohydride (102 mg, 1.62 mmol, 2.5 equivalents), react at 20 degrees Celsius for 1 hour. After the reaction was completed, the concentrated residue was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Synergi C18 150 × 25 millimeters × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 0%-30 %, 10 minutes) to isolate compound 47-4.
MS-ESI计算值[M+H] +363,实测值363。 MS-ESI calculated [M+H] + 363, found 363.
1HNMR(400MHz,CD 3OD)δ＝6.97-6.68(m,1H),6.49-6.16(m,1H),3.78(s,3H),3.54(br d,J＝5.1Hz,2H),3.26(d,J＝13.4Hz,2H),2.73(dd,J＝2.6,13.4Hz,2H),2.55-2.48(m,2H),2.36(d,J＝2.8Hz,3H)。 1 HNMR (400MHz, CD 3 OD) δ = 6.97-6.68 (m, 1H), 6.49-6.16 (m, 1H), 3.78 (s, 3H), 3.54 (br d, J = 5.1Hz, 2H), 3.26 (d, J=13.4Hz, 2H), 2.73 (dd, J=2.6, 13.4Hz, 2H), 2.55-2.48 (m, 2H), 2.36 (d, J=2.8Hz, 3H).
第五步the fifth step
将化合物47-4(0.2克，552微摩尔，1当量)溶于甲醇(2毫升)，水(2毫升)，接着向反应液中加入碳酸钾(381毫克，2.76毫摩尔，5当量)，在60摄氏度下反应2小时。反应完成后，反应液过滤，滤液浓缩的残渣经高效液相色谱(色谱柱：Phenomenex luna C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：1％-20％，10分钟)分离得到化合物47-5。Compound 47-4 (0.2 g, 552 micromole, 1 eq) was dissolved in methanol (2 ml), water (2 ml), then potassium carbonate (381 mg, 2.76 mmol, 5 eq) was added to the reaction solution, React at 60°C for 2 hours. After the reaction was completed, the reaction solution was filtered, and the concentrated residue of the filtrate was subjected to high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 25 millimeters × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B% : 1%-20%, 10 min) to isolate compound 47-5.
MS-ESI计算值[M+H] +267,实测值267。 MS-ESI calculated [M+H] +267 , found 267.
1HNMR(400MHz,CD 3OD)δ＝6.44(dd,J＝3.1,10.0Hz,1H),6.23(dd,J＝3.1,9.4Hz,1H),4.33(br s,2H),4.05(br s,2H),3.77(s,3H),2.89(s,3H),2.82(br d,J＝13.2Hz,2H),2.67-2.56(m,2H)。 1 HNMR (400MHz, CD 3 OD) δ = 6.44 (dd, J = 3.1, 10.0Hz, 1H), 6.23 (dd, J = 3.1, 9.4Hz, 1H), 4.33 (br s, 2H), 4.05 (br s, 2H), 3.77 (s, 3H), 2.89 (s, 3H), 2.82 (br d, J=13.2Hz, 2H), 2.67-2.56 (m, 2H).
第六步step six
将化合物47-5(66毫克，248微摩尔，1当量)，化合物E(94毫克，297微摩尔，1.2当量)溶于二氧六环(3毫升)，接着向反应液中加入碳酸铯(323毫克，991微摩尔，4当量)，三(二亚苄基丙酮)二钯(22.7毫克，24.8微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(14.3毫克，24.8微摩尔，0.1当量)，在110摄氏度氮气保护下反应12小时。反应完成后，反应液过滤，滤液浓缩的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝8：1)纯化得到化合物47-6。Compound 47-5 (66 mg, 248 μmol, 1 eq), Compound E (94 mg, 297 μmol, 1.2 eq) were dissolved in dioxane (3 ml), and cesium carbonate ( 323 mg, 991 micromoles, 4 equiv), tris(dibenzylideneacetone)dipalladium (22.7 mg, 24.8 micromoles, 0.1 equiv), 4,5-bis(diphenylphosphine)-9,9-bis Methylxanthene (14.3 mg, 24.8 micromole, 0.1 eq.) was reacted at 110°C under nitrogen protection for 12 hours. After the reaction was completed, the reaction solution was filtered, and the residue from the concentrated filtrate was purified by silica gel thin-layer chromatography (dichloromethane:methanol=8:1) to obtain compound 47-6.
MS-ESI计算值[M+H] +546,实测值546。 MS-ESI calculated [M+H] + 546, found 546.
第七步step seven
将化合物47-6(20毫克，36.6微摩尔，1当量)溶于二氯甲烷(1毫升)，接着在0摄氏度下向反应液中加入二乙氨基三氟化硫(7.09毫克，44.0微摩尔，1.2当量)，在20摄氏度下反应2小时。反应完成后，加入饱和碳酸氢钠水溶液(2毫升)淬灭，反应液过滤，滤液浓缩的粗品经高效液相色谱(色谱柱：Phenomenex luna C18 150×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：15％-45％，10分钟)分离得到化合物47。Compound 47-6 (20 mg, 36.6 micromole, 1 eq) was dissolved in dichloromethane (1 mL), and diethylaminosulfur trifluoride (7.09 mg, 44.0 micromole , 1.2 equivalents), reacted at 20 degrees Celsius for 2 hours. After the reaction was completed, add saturated aqueous sodium bicarbonate solution (2 milliliters) to quench, the reaction solution was filtered, and the crude product concentrated by the filtrate was subjected to high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 25 millimeters × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 15%-45%, 10 minutes) isolated to obtain compound 47.
MS-ESI计算值[M+H] +548,实测值548。 MS-ESI calculated value [M+H] + 548, found value 548.
1HNMR(400MHz,CD 3OD)δ＝8.55(s,1H),8.24(dd,J＝4.3,8.3Hz,1H),8.20-8.16(m,1H),7.99-7.83(m,1H),7.75-7.54(m,2H),7.36-7.23(m,1H),6.76-6.50(m,1H),4.67-4.55(m,1H),4.21-4.03(m, 2H),3.83-3.73(m,4H),3.24-2.99(m,2H),2.98-2.88(m,2H),2.87-2.65(m,3H),1.86(s,3H),1.83(s,3H)。 1 HNMR (400MHz, CD 3 OD) δ＝8.55(s, 1H), 8.24(dd, J＝4.3, 8.3Hz, 1H), 8.20-8.16(m, 1H), 7.99-7.83(m, 1H), 7.75-7.54(m,2H),7.36-7.23(m,1H),6.76-6.50(m,1H),4.67-4.55(m,1H),4.21-4.03(m,2H),3.83-3.73(m ,4H), 3.24-2.99(m,2H), 2.98-2.88(m,2H), 2.87-2.65(m,3H), 1.86(s,3H), 1.83(s,3H).
实施例48Example 48
Figure PCTCN2022129958-appb-000095
Figure PCTCN2022129958-appb-000095
第一步first step
将化合物26-1(250毫克，518.98微摩尔，1.0当量)溶于甲苯(8毫升)和水(1毫升)，氮气置换保护，加入化合物L(236.01毫克，778.48微摩尔，1.5当量)，碳酸铯(507.29毫克，1.56毫摩尔，3当量)，正丁基-二(1-金刚烷基)膦(37.22毫克，103.80微摩尔，0.2当量)，醋酸钯(23.30毫克，103.80微摩尔，0.2当量)，氮气置换保护，反应液加热到120摄氏下反应16小时。反应完成后，过滤，滤液用乙酸乙酯(100mL)稀释，饱和食盐水(100mL)洗涤。有机相用无水硫酸钠干燥，过滤，滤液经减压浓缩得到的残渣经硅胶薄层色谱板(二氯甲烷：甲醇＝10:1)和高效液相色谱(色谱柱：Phenomenex luna C18 150×40毫米×15微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:56％-86％,10分钟)纯化得到化合物48-1。Compound 26-1 (250 mg, 518.98 micromoles, 1.0 equivalents) was dissolved in toluene (8 ml) and water (1 ml), nitrogen displacement protection was added, compound L (236.01 mg, 778.48 micromoles, 1.5 equivalents), carbonic acid Cesium (507.29 mg, 1.56 mmol, 3 equiv), n-butyl-bis(1-adamantyl)phosphine (37.22 mg, 103.80 micromol, 0.2 equiv), palladium acetate (23.30 mg, 103.80 micromol, 0.2 equiv ), nitrogen replacement protection, and the reaction solution was heated to 120 degrees Celsius for 16 hours. After the reaction was completed, it was filtered, and the filtrate was diluted with ethyl acetate (100 mL), and washed with saturated brine (100 mL). The organic phase was dried with anhydrous sodium sulfate, filtered, and the residue obtained by concentrating the filtrate under reduced pressure was subjected to silica gel thin-layer chromatography (dichloromethane:methanol=10:1) and high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150× 40 mm × 15 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 56%-86%, 10 minutes) purified to obtain compound 48-1.
MS-ESI计算值[M+H] +598,实测值598。 MS-ESI calculated value [M+H] + 598, found value 598.
第二步second step
将化合物48-1(89毫克，148.81微摩尔，1.0当量)溶于乙酸乙酯(2毫升)，加入硫酸(74.47毫克，744.06微摩尔，5.0当量)，反应液在20摄氏下反应0.5小时。反应完成后，反应液经减压浓缩得到粗品化合物48-2(硫酸盐)。Compound 48-1 (89 mg, 148.81 micromol, 1.0 equiv) was dissolved in ethyl acetate (2 ml), sulfuric acid (74.47 mg, 744.06 micromol, 5.0 equiv) was added, and the reaction solution was reacted at 20°C for 0.5 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain crude compound 48-2 (sulfate).
MS-ESI计算值[M+H] +498,实测值498。 MS-ESI calculated [M+H] + 498, found 498.
第三步third step
将化合物48-2(88毫克，硫酸盐，147.64微摩尔，1.0当量)溶于甲醇(3毫升)，加入N，N-二异丙基乙胺(57.25毫克，442.93微摩尔，3.0当量)，甲醛(37％，67.17毫克，738.21微摩尔，5当量)，氰基硼氢化钠(27.83毫克，442.93微摩尔，3.0当量)，反应液在20摄氏下反应1小时。反应完成后，反应液经减压浓缩得到的残渣经高效液相色谱(色谱柱：Unisil 3-100 C18 Ultra 150×50毫米×3微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:7％-37％，7分钟)纯化得到化合物48(甲酸盐)。Compound 48-2 (88 mg, sulfate salt, 147.64 μmol, 1.0 eq) was dissolved in methanol (3 mL), N,N-diisopropylethylamine (57.25 mg, 442.93 μmol, 3.0 eq) was added, Formaldehyde (37%, 67.17 mg, 738.21 micromoles, 5 equivalents), sodium cyanoborohydride (27.83 mg, 442.93 micromoles, 3.0 equivalents), and the reaction solution was reacted at 20 degrees Celsius for 1 hour. After the reaction was completed, the residue obtained by concentrating the reaction solution under reduced pressure was subjected to high performance liquid chromatography (chromatographic column: Unisil 3-100 C18 Ultra 150 × 50 mm × 3 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: Acetonitrile; B%: 7%-37%, 7 minutes) was purified to obtain compound 48 (formate salt).
MS-ESI计算值[M+H] +512,实测值512。 MS-ESI calculated value [M+H] + 512, found value 512.
1H NMR(400MHz,CD 3OD)δ＝8.35-8.25(m,1H),8.16-8.06(m,1H),7.85-7.77(m,1H),7.69-7.59(m,1H),7.53(m,1H),7.34-7.23(m,1H),7.01-6.92(m,2H),4.33(s,2H),4.09(s,2H),3.78-3.67(m, 4H),2.90(s,3H),2.80-2.65(m,2H),2.50-2.33(m,2H),1.87(s,3H),1.84(s,3H)。 1 H NMR (400MHz, CD 3 OD) δ=8.35-8.25 (m, 1H), 8.16-8.06 (m, 1H), 7.85-7.77 (m, 1H), 7.69-7.59 (m, 1H), 7.53 ( m,1H),7.34-7.23(m,1H),7.01-6.92(m,2H),4.33(s,2H),4.09(s,2H),3.78-3.67(m, 4H),2.90(s, 3H), 2.80-2.65(m,2H), 2.50-2.33(m,2H), 1.87(s,3H), 1.84(s,3H).
实施例49Example 49
Figure PCTCN2022129958-appb-000096
Figure PCTCN2022129958-appb-000096
第一步first step
将化合物49-1(2克，8.44毫摩尔，1当量)，化合物49-2(790.34毫克，10.13毫摩尔，1.2当量)，磷酸钾(2.69克，12.66毫摩尔，1.5当量)，4，5-双二苯基膦-9，9-二甲基氧杂氧杂蒽杂蒽(976.52毫克，1.69毫摩尔，0.2当量)和醋酸钯(189.45毫克，843.84微摩尔，0.1当量)溶于N，N-二甲基甲酰胺(20毫升)，置换三次氮气，并在氮气氛围下在100摄氏度下反应12小时。反应完成后，反应液经过滤，浓缩得到粗品。粗品经硅胶柱层析(石油醚：乙酸乙酯＝30：1-1：4)分离纯化得到化合物49-3。Compound 49-1 (2 grams, 8.44 mmol, 1 equivalent), compound 49-2 (790.34 mg, 10.13 mmol, 1.2 equivalents), potassium phosphate (2.69 grams, 12.66 mmol, 1.5 equivalents), 4,5 - Bisdiphenylphosphine-9,9-dimethyloxaxanthene (976.52 mg, 1.69 mmol, 0.2 equiv) and palladium acetate (189.45 mg, 843.84 micromol, 0.1 equiv) in N, N-dimethylformamide (20 ml) was replaced with nitrogen three times, and reacted at 100° C. for 12 hours under nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered and concentrated to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=30:1-1:4) to obtain compound 49-3.
MS-ESI计算值[M+H] +188,实测值188. MS-ESI calculated value [M+H] + 188, measured value 188.
1H NMR(400MHz,CD 3OD)δ＝7.25-7.02(m,2H),6.70(ddt,J＝3.6,4.5,7.9Hz,1H),1.83(d,J＝13.5Hz,6H)。 1 H NMR (400 MHz, CD 3 OD) δ=7.25-7.02 (m, 2H), 6.70 (ddt, J=3.6, 4.5, 7.9 Hz, 1H), 1.83 (d, J=13.5 Hz, 6H).
第二步second step
将化合物49-3(350毫克，1.87毫摩尔，1当量)溶于四氢呋喃(5毫升)，置换三次氮气，在0摄氏度下加入钠氢(112.2毫克，2.81毫摩尔，60％纯度，1.5当量)，搅拌半小时后，在0摄氏度下加入化合物49-4(514.54毫克，2.81毫摩尔，1.5当量)，并在氮气氛围下在0摄氏度下反应1小时。反应完成后，在25摄氏度下，反应液用饱和氯化铵溶液(50毫升)淬灭反应，加入乙酸乙酯(50毫升)稀释，再用乙酸乙酯(40毫升×3)萃取。有机相经无水硫酸钠干燥后，过滤浓缩得到粗品。粗品经高效液相色谱(色谱柱：Phenomenex Luna C18 150×40毫米×15微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％：20％-50％，10分钟)分离得到化合物49-5。Compound 49-3 (350 mg, 1.87 mmol, 1 eq) was dissolved in tetrahydrofuran (5 ml), nitrogen was replaced three times, and sodium hydrogen (112.2 mg, 2.81 mmol, 60% purity, 1.5 eq) was added at 0°C , after stirring for half an hour, compound 49-4 (514.54 mg, 2.81 mmol, 1.5 equiv) was added at 0°C, and reacted at 0°C for 1 hour under nitrogen atmosphere. After the reaction was completed, the reaction solution was quenched with saturated ammonium chloride solution (50 mL) at 25°C, diluted with ethyl acetate (50 mL), and extracted with ethyl acetate (40 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was separated by high-performance liquid chromatography (chromatographic column: Phenomenex Luna C18 150 × 40 mm × 15 microns; mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 20%-50%, 10 minutes) separation Compound 49-5 was obtained.
第三步third step
将化合物49-5(100毫克，299.3微摩尔，1当量)，化合物4-2(89.19毫克，359.16微摩尔，1.2当量)，碳酸铯(292.55毫克，897.9微摩尔，3当量)，4，5-双二苯基膦-9，9-二甲基氧杂氧杂蒽杂蒽(34.64毫克，59.86微摩尔，0.2当量)和三(二亚苄基丙酮)二钯(27.41毫克，29.93微摩尔，0.1当量)溶于二氧六环(5毫升)，置换三次氮气，并在氮气氛围下在105摄氏度下反应12小时。反应完成后，反应液经过滤浓缩得到粗品。粗品经硅胶薄层色谱板(乙酸乙酯：甲醇＝20：1)纯化。再经高效液相色谱(色谱柱：Phenomenex Luna C18 150×25毫米×10微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％：30％-60％，10分钟)分离得到化合物49。Compound 49-5 (100 mg, 299.3 micromoles, 1 equivalent), compound 4-2 (89.19 mg, 359.16 micromoles, 1.2 equivalents), cesium carbonate (292.55 mg, 897.9 micromoles, 3 equivalents), 4,5 - bis-diphenylphosphine-9,9-dimethyloxaxanthene (34.64 mg, 59.86 micromoles, 0.2 equivalents) and tris(dibenzylideneacetone) dipalladium (27.41 mg, 29.93 micromoles , 0.1 eq) was dissolved in dioxane (5 ml), nitrogen was replaced three times, and reacted at 105° C. for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel thin-layer chromatography (ethyl acetate:methanol=20:1). Then through high performance liquid chromatography (chromatographic column: Phenomenex Luna C18 150 * 25 millimeters * 10 microns; Mobile phase: mobile phase A: formic acid solution; Mobile phase B: acetonitrile; B%: 30%-60%, 10 minutes) separation Compound 49 was obtained.
MS-ESI计算值[M+H] +546,实测值546. MS-ESI calculated value [M+H] + 546, measured value 546.
1H NMR(400MHz,CD 3OD)δ＝8.13(s,1H),7.79(dd,J＝7.2,12.0Hz,1H),7.66(ddt,J＝2.9,4.9,7.9Hz,1H),7.59-7.51(m,1H),7.01(dd,J＝1.1,8.3Hz,1H),6.51(t,J＝8.2Hz,1H),6.09(dd,J＝1.3,8.1Hz,1H),3.90-3.83(m,1H),3.82(s,2H),3.77(s,2H),3.62(s,3H),3.24(s,3H),2.57-2.45(m,2H),2.15-2.03(m,2H),1.75(br d,J＝13.7Hz,6H)。 1 H NMR (400MHz, CD 3 OD) δ=8.13(s,1H),7.79(dd,J=7.2,12.0Hz,1H),7.66(ddt,J=2.9,4.9,7.9Hz,1H),7.59 -7.51(m,1H),7.01(dd,J=1.1,8.3Hz,1H),6.51(t,J=8.2Hz,1H),6.09(dd,J=1.3,8.1Hz,1H),3.90- 3.83(m,1H),3.82(s,2H),3.77(s,2H),3.62(s,3H),3.24(s,3H),2.57-2.45(m,2H),2.15-2.03(m, 2H), 1.75 (br d, J=13.7Hz, 6H).
实施例50Example 50
Figure PCTCN2022129958-appb-000097
Figure PCTCN2022129958-appb-000097
第一步first step
将化合物50-1(3克，12.66毫摩尔，1当量)，化合物49-2(1.19克，15.19毫摩尔，1.2当量)，磷酸钾(4.03克，12.66毫摩尔，1.5当量)，4，5-双二苯基膦-9，9-二甲基氧杂氧杂蒽杂蒽(1.46克，2.53毫摩尔，0.2当量)和醋酸钯(284.17毫克，1.27毫摩尔，0.1当量)溶于N,N-二甲基甲酰胺(50毫升)，置换三次氮气，并在氮气氛围下在100摄氏度下反应12小时。反应完成后，反应液加入水(20毫升)淬灭反应，用乙酸乙酯(50毫升)稀释，再用乙酸乙酯(50毫升×3)萃取，合并有机相用无水硫酸钠干燥，过滤，浓缩得到粗品。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1-1：4)分离纯化得到化合物50-2。Compound 50-1 (3 grams, 12.66 mmol, 1 equivalent), compound 49-2 (1.19 grams, 15.19 mmol, 1.2 equivalents), potassium phosphate (4.03 grams, 12.66 mmol, 1.5 equivalents), 4,5 -Bisdiphenylphosphine-9,9-dimethyloxaxanthene (1.46 g, 2.53 mmol, 0.2 eq) and palladium acetate (284.17 mg, 1.27 mmol, 0.1 eq) were dissolved in N, N-dimethylformamide (50 ml) was replaced with nitrogen three times, and reacted at 100° C. for 12 hours under nitrogen atmosphere. After the reaction was complete, the reaction solution was quenched by adding water (20 ml), diluted with ethyl acetate (50 ml), extracted with ethyl acetate (50 ml×3), and the combined organic phases were dried over anhydrous sodium sulfate and filtered. , concentrated to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1-1:4) to obtain compound 50-2.
MS-ESI计算值[M+H] +188,实测值188。 MS-ESI calculated value [M+H] + 188, found value 188.
第二步second step
将化合物50-2(300毫克，1.6毫摩尔，1当量)溶于乙腈(10毫升)，在25摄氏度下加入碳酸钾(664.63毫克，4.81毫摩尔，3当量)，搅拌半小时后，在25摄氏度下加入化合物49-4(441.04毫克，2.4毫摩尔，1.5当量)，并在80摄氏度下反应12小时。反应完成后，在25摄氏度下，反应液加入水(50毫升)淬灭反应，加入乙酸乙酯(50毫升)稀释，再用乙酸乙酯(100毫升×2)萃取，再用饱和食盐水(50毫升×2)洗涤。有机相经无水硫酸钠干燥后，过滤浓缩得到粗品。粗品经硅胶柱层析(石油醚：乙酸乙酯＝50：1-1：2)分离纯化得到化合物50-3。Compound 50-2 (300 mg, 1.6 mmol, 1 eq) was dissolved in acetonitrile (10 ml), and potassium carbonate (664.63 mg, 4.81 mmol, 3 eq) was added at 25 degrees Celsius, and after stirring for half an hour, at 25 Compound 49-4 (441.04 mg, 2.4 mmol, 1.5 equiv) was added at 100C and reacted at 800C for 12 hours. After the reaction was completed, at 25 degrees Celsius, the reaction solution was quenched by adding water (50 ml), diluted with ethyl acetate (50 ml), extracted with ethyl acetate (100 ml × 2), and then washed with saturated brine ( 50 ml × 2) wash. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 50:1-1:2) to obtain compound 50-3.
MS-ESI计算值[M+H] +334,实测值334。 MS-ESI calculated [M+H] + 334, found 334.
第三步third step
将化合物50-3(375毫克，1.12毫摩尔，1当量)，化合物4-2(306.58毫克，1.23毫摩尔，1.1当量)，碳酸铯(1.1克，3.37毫摩尔，3当量)，4，5-双二苯基膦-9，9-二甲基氧杂氧杂蒽杂蒽(129.89毫克，224.47微摩尔，0.2当量)和三(二亚苄基丙酮)二钯(102.78毫克，112.24微摩尔，0.1当量)溶于二氧六环(5毫升)，置换三次氮气，并在氮气氛围下在105摄氏度下反应12小时。反应完成后，反应液经过滤浓缩得到 粗品。粗品经硅胶柱层析(石油醚：乙酸乙酯＝20：1-0：1)纯化，再经高效液相色谱(色谱柱：Phenomenex Luna C18 150×25毫米×10微米；流动相：流动相A：甲酸溶液；流动相B：乙腈；B％：49％-79％，10分钟)分离得到化合物50。Compound 50-3 (375 mg, 1.12 mmol, 1 equivalent), compound 4-2 (306.58 mg, 1.23 mmol, 1.1 equivalent), cesium carbonate (1.1 g, 3.37 mmol, 3 equivalents), 4,5 - Bis-diphenylphosphine-9,9-dimethyloxaxanthene (129.89 mg, 224.47 micromoles, 0.2 equivalents) and tris(dibenzylideneacetone) dipalladium (102.78 mg, 112.24 micromoles , 0.1 eq) was dissolved in dioxane (5 ml), nitrogen was replaced three times, and reacted at 105° C. for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate=20:1-0:1), and then by high performance liquid chromatography (column: Phenomenex Luna C18 150 × 25 mm × 10 microns; mobile phase: mobile phase A: formic acid solution; mobile phase B: acetonitrile; B%: 49%-79%, 10 minutes) isolated to obtain compound 50.
MS-ESI计算值[M+H] +546,实测值546. MS-ESI calculated value [M+H] + 546, measured value 546.
1H NMR(400MHz,CD 3OD)δ＝8.53(dd,J＝3.4,8.6Hz,1H),8.11(s,1H),7.53-7.44(m,1H),7.41(dd,J＝1.4,8.1Hz,1H),6.99-6.85(m,2H),6.30(dd,J＝1.4,8.1Hz,1H),3.92-3.78(m,4H),3.67(s,3H),3.25(s,3H),2.67-2.46(m,2H),2.18-2.06(m,2H),1.95(dd,J＝2.4,13.9Hz,6H)。 1 H NMR (400MHz, CD 3 OD) δ = 8.53 (dd, J = 3.4, 8.6 Hz, 1H), 8.11 (s, 1H), 7.53-7.44 (m, 1H), 7.41 (dd, J = 1.4, 8.1Hz, 1H), 6.99-6.85(m, 2H), 6.30(dd, J=1.4, 8.1Hz, 1H), 3.92-3.78(m, 4H), 3.67(s, 3H), 3.25(s, 3H ), 2.67-2.46 (m, 2H), 2.18-2.06 (m, 2H), 1.95 (dd, J=2.4, 13.9Hz, 6H).
实施例51Example 51
Figure PCTCN2022129958-appb-000098
Figure PCTCN2022129958-appb-000098
第一步first step
将化合物51-1(2.2克，9.28毫摩尔，1当量)，化合物49-2(869.37毫克，11.14毫摩尔，1.2当量)，磷酸钾(2.96克，13.92毫摩尔，1.5当量)，4，5-双二苯基膦-9，9-二甲基氧杂氧杂蒽杂蒽(1.07克，1.86毫摩尔，0.2当量)和醋酸钯(208.39毫克，928.22微摩尔，0.1当量)溶于N，N-二甲基甲酰胺(22毫升)，置换三次氮气，并在氮气氛围下在100摄氏度下反应12小时。反应完成后，反应液经过滤，加入水(100毫升)稀释，再用乙酸乙酯(100毫升×2)萃取，饱和食盐水(100毫升)洗涤。有机相经无水硫酸钠干燥后，过滤浓缩得到粗品浓缩得到粗品。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1-0：1)分离纯化得到化合物51-2。Compound 51-1 (2.2 grams, 9.28 mmol, 1 equivalent), compound 49-2 (869.37 mg, 11.14 mmol, 1.2 equivalents), potassium phosphate (2.96 grams, 13.92 mmol, 1.5 equivalents), 4,5 - Bisdiphenylphosphine-9,9-dimethyloxaxanthene (1.07 g, 1.86 mmol, 0.2 eq) and palladium acetate (208.39 mg, 928.22 micromol, 0.1 eq) in N, N-dimethylformamide (22 ml) was replaced with nitrogen three times, and reacted at 100° C. for 12 hours under nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, diluted with water (100 mL), extracted with ethyl acetate (100 mL×2), and washed with saturated brine (100 mL). After the organic phase was dried over anhydrous sodium sulfate, the crude product was obtained by filtration and concentrated. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1-0:1) to obtain compound 51-2.
MS-ESI计算值[M+H] +188,实测值188。 MS-ESI calculated value [M+H] + 188, found value 188.
1H NMR(400MHz,CD 3OD)δ＝7.44-7.11(m,1H),6.80-6.08(m,2H),2.29-1.54(m,6H)。 1 H NMR (400 MHz, CD 3 OD) δ = 7.44-7.11 (m, 1H), 6.80-6.08 (m, 2H), 2.29-1.54 (m, 6H).
第二步second step
将化合物51-2(200毫克，1.07毫摩尔，1当量)溶于四氢呋喃(5毫升)，在0摄氏度下和氮气保护下加入钠氢(64.11毫克，1.60毫摩尔，60％纯度，1.5当量)，搅拌半小时后，在0摄氏度下加入化合物49-4(294.02毫克，1.60毫摩尔，1.5当量)，并在氮气氛围下在0摄氏度下反应1小时。反应完成后，在25摄氏度下，反应液用饱和氯化铵溶液(10毫升)淬灭反应，再用乙酸乙酯(10毫升×2)萃取，饱和食盐水(10毫升)洗涤。有机相经无水硫酸钠干燥后，过滤浓缩得到粗品。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1-20：1)分离纯化得到化合物51-3。Compound 51-2 (200 mg, 1.07 mmol, 1 eq) was dissolved in tetrahydrofuran (5 ml), and sodium hydrogen (64.11 mg, 1.60 mmol, 60% purity, 1.5 eq) was added at 0°C under nitrogen protection , after stirring for half an hour, compound 49-4 (294.02 mg, 1.60 mmol, 1.5 equiv) was added at 0°C, and reacted at 0°C for 1 hour under nitrogen atmosphere. After the reaction was completed, the reaction solution was quenched with saturated ammonium chloride solution (10 mL) at 25°C, extracted with ethyl acetate (10 mL×2), and washed with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=100:1-20:1) to obtain compound 51-3.
第三步third step
将化合物51-3(200毫克，598.60微摩尔，1当量)，化合物4-2(178.37毫克，718.32微摩尔，1.2当量)， 碳酸铯(585.11毫克，1.80毫摩尔，3当量)，4，5-双二苯基膦-9，9-二甲基氧杂氧杂蒽杂蒽(69.27毫克，119.72微摩尔，0.2当量)和三(二亚苄基丙酮)二钯(54.81毫克，59.86微摩尔，0.1当量)溶于二氧六环(5毫升)，置换三次氮气，并在氮气氛围下在105摄氏度下反应10小时。反应完成后，反应液经过滤浓缩得到粗品。粗品经经高效液相色谱(色谱柱：Unisil 3-100C18Ultra 150×50毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:44％-74％，10分钟)分离得到化合物51。Compound 51-3 (200 mg, 598.60 micromoles, 1 equivalent), compound 4-2 (178.37 mg, 718.32 micromoles, 1.2 equivalents), cesium carbonate (585.11 mg, 1.80 mmoles, 3 equivalents), 4,5 - Bis-diphenylphosphine-9,9-dimethyloxaxanthene (69.27 mg, 119.72 micromoles, 0.2 equivalents) and tris(dibenzylideneacetone) dipalladium (54.81 mg, 59.86 micromoles , 0.1 eq) was dissolved in dioxane (5 ml), nitrogen was replaced three times, and reacted at 105° C. for 10 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered and concentrated to obtain a crude product. The crude product was subjected to high performance liquid chromatography (chromatographic column: Unisil 3-100C18Ultra 150 × 50 mm × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 44%-74%, 10 minutes ) to isolate compound 51.
MS-ESI计算值[M+H] +546，实测值546。 MS-ESI calculated [M+H] + 546, found 546.
1HNMR(400MHz,CD 3OD)δ＝8.43-8.35(m,1H),8.12(s,1H),7.65-7.55(m,1H),7.39-7.32(m,1H),6.98-6.86(m,2H),6.31-6.18(m,1H),3.91-3.78(m,4H),3.66(s,3H),3.23(s,3H),2.57-2.46(m,2H),2.13-2.02(m,2H),1.88(s,3H),1.84(s,3H)。 1 HNMR (400MHz, CD 3 OD) δ = 8.43-8.35(m, 1H), 8.12(s, 1H), 7.65-7.55(m, 1H), 7.39-7.32(m, 1H), 6.98-6.86(m ,2H),6.31-6.18(m,1H),3.91-3.78(m,4H),3.66(s,3H),3.23(s,3H),2.57-2.46(m,2H),2.13-2.02(m ,2H), 1.88(s,3H), 1.84(s,3H).
实施例52Example 52
Figure PCTCN2022129958-appb-000099
Figure PCTCN2022129958-appb-000099
第一步first step
将化合物52-1(1克，4.22毫摩尔，1当量)溶于N，N二甲基甲酰胺(15毫升)，接着向反应液中加入化合物49-2(395毫克，5.06毫摩尔，1.2当量)，磷酸钾(1.34克，6.33毫摩尔，1.5当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(488毫克，844微摩尔，0.2当量)，醋酸钯(94.72毫克，422微摩尔，0.1当量)，在120摄氏度氮气保护下反应12小时。反应完成后，反应液过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-0：1)纯化得到化合物52-2。Compound 52-1 (1 g, 4.22 mmol, 1 eq) was dissolved in N,N dimethylformamide (15 ml), then compound 49-2 (395 mg, 5.06 mmol, 1.2 equiv), potassium phosphate (1.34 g, 6.33 mmol, 1.5 equiv), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (488 mg, 844 micromol, 0.2 equiv) , palladium acetate (94.72 mg, 422 micromole, 0.1 eq), reacted at 120 degrees centigrade under nitrogen protection for 12 hours. After the reaction was completed, the reaction solution was filtered, and the residue concentrated under reduced pressure was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-0:1) to obtain compound 52-2.
MS-ESI计算值[M+H] +188,实测值188。 MS-ESI calculated value [M+H] + 188, found value 188.
1HNMR(400MHz,CD 3OD)δ＝7.08-6.99(m,2H),6.79-6.71(m,1H),1.83-1.80(m,3H),1.75(br s,3H)。 1 H NMR (400 MHz, CD 3 OD) δ = 7.08-6.99 (m, 2H), 6.79-6.71 (m, 1H), 1.83-1.80 (m, 3H), 1.75 (br s, 3H).
第二步second step
将化合物52-2(330毫克，1.76毫摩尔，1当量)溶于N，N二甲基甲酰胺(5毫升)，接着加入碳酸钾(731毫克，5.29毫摩尔，3当量)，在25摄氏度下反应0.5小时。然后接着加入化合物49-4(388毫克，2.12毫摩尔，1.2当量)，在60摄氏度下反应12小时。反应完成后，加水(60毫升)稀释，乙酸乙酯萃取(30毫升×3)，合并的有机相用饱和食盐水溶液(40毫升×3)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩。粗品经硅胶薄层色谱板(石油醚：乙酸乙酯＝0：1)纯化得到化合物52-3。Compound 52-2 (330 mg, 1.76 mmol, 1 eq) was dissolved in N,N dimethylformamide (5 ml), followed by addition of potassium carbonate (731 mg, 5.29 mmol, 3 eq), at 25° C. The reaction was carried out for 0.5 hours. Then compound 49-4 (388mg, 2.12mmol, 1.2eq) was added and reacted at 60°C for 12 hours. After the reaction was complete, dilute with water (60 ml), extract with ethyl acetate (30 ml × 3), wash the combined organic phase with saturated saline solution (40 ml × 3), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate . The crude product was purified by silica gel thin-layer chromatography (petroleum ether: ethyl acetate = 0:1) to obtain compound 52-3.
MS-ESI计算值[M+H] +334,实测值334。 MS-ESI calculated [M+H] + 334, found 334.
第三步third step
将化合物52-3(150毫克，449微摩尔，1当量)，化合物4-2(134毫克，539微摩尔，1.2当量)溶于二氧六环(4毫升)，接着向反应液中加入碳酸铯(439毫克，1.35毫摩尔，3当量)，三(二亚苄基丙酮)二钯(41毫克，45微摩尔，0.1当量)，4,5-双(二苯基膦)-9,9-二甲基氧杂蒽(52毫克，89.8微摩尔，0.2当量)，在氮气保护下105摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩的粗品经硅胶薄层色谱板(乙酸乙酯：甲醇＝10：1)纯化后的残渣经高效液相色谱(色谱柱：Phenomenex C18 75×30毫米×3微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:38％-68％，7分钟)分离得到化合物52。Compound 52-3 (150 mg, 449 micromoles, 1 equivalent), compound 4-2 (134 mg, 539 micromoles, 1.2 equivalents) were dissolved in dioxane (4 ml), and then carbonic acid was added to the reaction solution. Cesium (439 mg, 1.35 mmol, 3 equiv), Tris(dibenzylideneacetone)dipalladium (41 mg, 45 micromol, 0.1 equiv), 4,5-bis(diphenylphosphine)-9,9 -Dimethylxanthene (52 mg, 89.8 micromole, 0.2 eq), reacted at 105° C. for 12 hours under nitrogen protection. After the reaction was completed, the reaction solution was filtered, and the concentrated crude product of the filtrate was purified by silica gel thin-layer chromatography (ethyl acetate: methanol=10: 1), and the residue after purification by high-performance liquid chromatography (chromatographic column: Phenomenex C18 75 × 30 mm × 3 Micron; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 38%-68%, 7 minutes) isolated to obtain compound 52.
MS-ESI计算值[M+H] +546,实测值546。 MS-ESI calculated [M+H] + 546, found 546.
1HNMR(400MHz,CD 3OD)δ＝8.29-8.18(m,1H),8.11(s,1H),7.53-7.42(m,1H),7.35-7.26(m,2H),6.86-6.76(m,1H),6.31-6.20(m,1H),4.66-4.53(m,1H),3.89-3.86(m,2H),3.83-3.80(m,2H),3.67-3.62(m,3H),3.27-3.22(m,3H),2.58-2.47(m,2H),2.11(ddd,J＝2.9,6.9,9.8Hz,2H),1.87(s,3H),1.84(s,3H)。 1 HNMR (400MHz, CD 3 OD) δ=8.29-8.18(m,1H),8.11(s,1H),7.53-7.42(m,1H),7.35-7.26(m,2H),6.86-6.76(m ,1H),6.31-6.20(m,1H),4.66-4.53(m,1H),3.89-3.86(m,2H),3.83-3.80(m,2H),3.67-3.62(m,3H),3.27 -3.22 (m, 3H), 2.58-2.47 (m, 2H), 2.11 (ddd, J=2.9, 6.9, 9.8Hz, 2H), 1.87 (s, 3H), 1.84 (s, 3H).
实施例53Example 53
Figure PCTCN2022129958-appb-000100
Figure PCTCN2022129958-appb-000100
第一步first step
将化合物B-1(1克，4.69毫摩尔，1当量)，碘化亚酮(178.6毫克，937.77微摩尔，0.2当量)溶于乙腈(12毫升)中，接着在50摄氏度加入溶于乙腈(6毫升)的2-(氟代磺酰基)-二氟乙酸(1.25克，7.03毫摩尔，728.22微升，1.5当量)，在50摄氏度下反应1小时。反应完成后，将反应液过滤，滤液减压浓缩，浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-10：1)纯化得到得到化合物53-1。Compound B-1 (1 gram, 4.69 mmol, 1 eq), ketone iodide (178.6 mg, 937.77 micromol, 0.2 eq) were dissolved in acetonitrile (12 ml), and then added in acetonitrile ( 6 ml) of 2-(fluorosulfonyl)-difluoroacetic acid (1.25 g, 7.03 mmol, 728.22 μl, 1.5 equiv), reacted at 50°C for 1 hour. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated under reduced pressure, and the concentrated residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-10:1) to obtain compound 53-1.
1H NMR(400MHz,CD 3OD)δ＝6.63-6.00(m,1H),4.57-4.44(m,1H),4.09-3.69(m,4H),2.73-2.42(m,2H),2.31(ddd,J＝3.0,7.2,10.1Hz,2H),1.45-1.27(m,9H). 1 H NMR (400MHz, CD 3 OD) δ = 6.63-6.00 (m, 1H), 4.57-4.44 (m, 1H), 4.09-3.69 (m, 4H), 2.73-2.42 (m, 2H), 2.31 ( ddd,J＝3.0,7.2,10.1Hz,2H),1.45-1.27(m,9H).
第二步second step
将化合物53-1(220毫克，835.61微摩尔，1当量)溶于二氯甲烷(3毫升)和三氟乙酸(1毫升)，在25摄氏度下反应1小时。反应完成后，将反应液过滤，滤液减压浓缩得到化合物53-2(三氟乙酸盐)。Compound 53-1 (220 mg, 835.61 μmol, 1 equiv) was dissolved in dichloromethane (3 mL) and trifluoroacetic acid (1 mL), and reacted at 25° C. for 1 hour. After the reaction was completed, the reaction liquid was filtered, and the filtrate was concentrated under reduced pressure to obtain compound 53-2 (trifluoroacetic acid salt).
第三步third step
将化合物53-2(105.02毫克，78.88微摩尔，1当量，三氟乙酸盐)，化合物26-1(182.51毫克，378.88微摩尔，1当量)溶于二氧六环(6毫升)，接着向反应液中加入碳酸铯(432.06毫克，1.33毫摩尔，3.5当量)，(±)-2，2-双(二苯膦基)-1，1-联萘(47.18毫克，75.78微摩尔，0.2当量)，醋酸钯(8.51毫克， 37.89微摩尔，0.1当量)，氮气置换三次，在110摄氏度下反应12小时。反应完成后，将反应液过滤，滤液用水(20毫升)稀释，用乙酸乙酯(20毫升×2)萃取，有机相用饱和的食盐水(20毫升)洗涤，无水硫酸钠干燥，过滤，浓缩的残渣经制备高效液相色谱法(色谱柱：Phenomenex C18 75毫米×30毫米×3微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：42％-72％，7分钟)分离得到化合物53。Compound 53-2 (105.02 mg, 78.88 micromol, 1 equiv, trifluoroacetate), compound 26-1 (182.51 mg, 378.88 micromol, 1 equiv) were dissolved in dioxane (6 ml), followed by Cesium carbonate (432.06 mg, 1.33 mmol, 3.5 equivalents), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthalene (47.18 mg, 75.78 micromol, 0.2 equivalent), palladium acetate (8.51 mg, 37.89 micromole, 0.1 equivalent), replaced by nitrogen three times, and reacted at 110 degrees Celsius for 12 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was diluted with water (20 ml), extracted with ethyl acetate (20 ml×2), the organic phase was washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and filtered. The concentrated residue was subjected to preparative high performance liquid chromatography (chromatographic column: Phenomenex C18 75 mm × 30 mm × 3 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 42%-72%, 7 min) to isolate compound 53.
MS-ESI计算值[M+H] +564，实测值564。 MS-ESI calculated [M+H] + 564, found 564.
1H NMR(400MHz,CD 3OD)δ＝8.34(dd,J＝4.5,8.1Hz,1H),8.11(s,1H),7.70-7.51(m,2H),7.45-7.27(m,2H),6.82(t,J＝8.1Hz,1H),6.55-6.09(m,2H),4.59(br d,J＝7.1Hz,1H),3.87(d,J＝19.0Hz,4H),3.67(s,3H),2.67-2.54(m,2H),2.43-2.25(m,2H),1.86(d,J＝13.6Hz,6H). 1 H NMR (400MHz, CD 3 OD) δ=8.34(dd,J=4.5,8.1Hz,1H),8.11(s,1H),7.70-7.51(m,2H),7.45-7.27(m,2H) ,6.82(t,J=8.1Hz,1H),6.55-6.09(m,2H),4.59(br d,J=7.1Hz,1H),3.87(d,J=19.0Hz,4H),3.67(s ,3H),2.67-2.54(m,2H),2.43-2.25(m,2H),1.86(d,J=13.6Hz,6H).
实施例54Example 54
Figure PCTCN2022129958-appb-000101
Figure PCTCN2022129958-appb-000101
第一步first step
将化合物54-1(1克，6.28毫摩尔，1当量)溶于N，N-二甲基甲酰胺(10毫升)，接着在0摄氏度下向反应液中加入N-溴代丁二酰亚胺(1.12克，6.28毫摩尔，1当量)，在25摄氏度下反应1小时。反应完成后，加水(200毫升)稀释，用饱和的碳酸氢钠水溶液调节pH到8，乙酸乙酯萃取(200毫升×2)，合并的有机相用饱和食盐水溶液(200毫升)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-10：1)纯化得到得到化合物54-2。Compound 54-1 (1 g, 6.28 mmol, 1 eq) was dissolved in N, N-dimethylformamide (10 ml), then N-bromosuccinyl was added to the reaction solution at 0°C Amine (1.12 g, 6.28 mmol, 1 equiv), reacted at 25 °C for 1 hour. After completion of the reaction, dilute with water (200 ml), adjust the pH to 8 with saturated aqueous sodium bicarbonate, extract with ethyl acetate (200 ml × 2), and wash the combined organic phase with saturated aqueous salt solution (200 ml). After drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-10:1) to obtain compound 54-2.
MS-ESI计算值[M+H] +238，实测值238。 MS-ESI calculated [M+H] +238 , found 238.
1H NMR(400MHz,CD 3OD)δ＝7.98(s,1H),6.43(dd,J＝2.1,10.6Hz,1H),3.81(d,J＝0.7Hz,3H). 1 H NMR (400MHz, CD 3 OD) δ=7.98(s, 1H), 6.43(dd, J=2.1, 10.6Hz, 1H), 3.81(d, J=0.7Hz, 3H).
第二步second step
将化合物53-2(2克，8.4毫摩尔，1当量)，化合物E(2.66克，8.4毫摩尔，1当量)溶于二氧六环(20毫升)，接着向反应液中加入甲烷磺酸(807.55毫克，8.4毫摩尔，598.18微升，1当量)，氮气置换三次，在100摄氏度下反应12小时。反应完成后，加水(20毫升)稀释，然后用4摩尔每升的氢氧化钠水溶液调节pH至8，乙酸乙酯萃取(20毫升×2)，合并的有机相用饱和食盐水溶液(20毫升)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩得到的残渣用乙酸乙酯(20毫升)在25摄氏度下打浆30分钟，得到化合物54-3。Compound 53-2 (2 g, 8.4 mmol, 1 eq), compound E (2.66 g, 8.4 mmol, 1 eq) were dissolved in dioxane (20 ml), then methanesulfonic acid was added to the reaction solution (807.55 mg, 8.4 mmol, 598.18 microliters, 1 equiv), replaced with nitrogen three times, and reacted at 100 degrees Celsius for 12 hours. After completion of the reaction, add water (20 ml) to dilute, then adjust the pH to 8 with 4 moles of aqueous sodium hydroxide solution per liter, extract with ethyl acetate (20 ml × 2), and combine the organic phases with saturated saline solution (20 ml) After washing, the organic phase was dried over anhydrous sodium sulfate, filtered, and the concentrated residue was slurried with ethyl acetate (20 mL) at 25°C for 30 minutes to obtain compound 54-3.
MS-ESI计算值[M+H] +519，实测值519。 MS-ESI calculated [M+H] + 519, found 519.
第三步third step
将化合物54-3(500毫克，965.83微摩尔，1当量)，化合物A(173.86毫克，1.06毫摩尔，1.1当量，盐酸盐)溶于二氧六环(10毫升)，接着向反应液中加入叔丁醇钠(324.86毫克，3.38毫摩尔，3.5当量)，(±)-2，2-双(二苯膦基)-1，1-联萘(120.28毫克，193.17微摩尔，0.2当量)，醋酸钯(21.68毫克，96.58微摩尔，0.1当量)，氮气置换三次，在110摄氏度下反应12小时。反应完成后加水(20毫升)稀释，乙酸乙酯萃取(20毫升×2)，合并的有机相用饱和食盐水溶液(20毫升)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩的残渣经高效液相色谱(色谱柱：YMC Triart C18 150毫米×25毫米×5微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：56％-86％，10分钟)分离得到化合物54。Compound 54-3 (500 mg, 965.83 micromole, 1 eq), compound A (173.86 mg, 1.06 mmol, 1.1 eq, hydrochloride) were dissolved in dioxane (10 ml), and then added to the reaction solution Add sodium tert-butoxide (324.86 mg, 3.38 mmol, 3.5 equiv), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (120.28 mg, 193.17 micromol, 0.2 equiv) , palladium acetate (21.68 mg, 96.58 micromole, 0.1 eq), nitrogen replacement three times, reacted at 110 degrees Celsius for 12 hours. After the reaction was completed, it was diluted with water (20 ml), extracted with ethyl acetate (20 ml × 2), the combined organic phase was washed with saturated saline solution (20 ml), the organic phase was dried over anhydrous sodium sulfate, filtered, and the concentrated residue was washed with High performance liquid chromatography (chromatographic column: YMC Triart C18 150 millimeters * 25 millimeters * 5 microns; Mobile phase: mobile phase A: formic acid aqueous solution; Mobile phase B: acetonitrile; B%: 56%-86%, 10 minutes) separates and obtains Compound 54.
MS-ESI计算值[M+H] +564，实测值564。 MS-ESI calculated [M+H] + 564, found 564.
1H NMR(400MHz,CD 3OD)δ＝8.25(dd,J＝4.5,8.1Hz,1H),8.09(s,1H),7.68-7.45(m,3H),7.35-7.24(m,1H),4.07(t,J＝2.3Hz,2H),4.02(t,J＝2.3Hz,2H),3.88-3.77(m,4H),3.23(s,3H),2.52(br d,J＝3.1Hz,2H),2.14-2.02(m,2H),1.84(d,J＝13.6Hz,6H). 1 H NMR (400MHz, CD 3 OD) δ=8.25(dd,J=4.5,8.1Hz,1H),8.09(s,1H),7.68-7.45(m,3H),7.35-7.24(m,1H) ,4.07(t,J=2.3Hz,2H),4.02(t,J=2.3Hz,2H),3.88-3.77(m,4H),3.23(s,3H),2.52(br d,J=3.1Hz ,2H),2.14-2.02(m,2H),1.84(d,J＝13.6Hz,6H).
实施例55Example 55
Figure PCTCN2022129958-appb-000102
Figure PCTCN2022129958-appb-000102
第一步first step
将化合物55-1(5克，35.43毫摩尔，1当量)溶于N，N-二甲基甲酰胺(100毫升)，接着在0摄氏度下向反应液中加入N-溴代丁二酰亚胺(6.31克，35.43毫摩尔，1当量)，在25摄氏度下反应1小时。反应完成后，加水(200毫升)稀释，用饱和的碳酸氢钠水溶液调pH到8，乙酸乙酯萃取(200毫升×2)，合并的有机相用饱和食盐水溶液(200毫升)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-10：1)纯化得到化合物55-2。Compound 55-1 (5 g, 35.43 mmol, 1 eq) was dissolved in N, N-dimethylformamide (100 ml), and then N-bromosuccinyl was added to the reaction solution at 0°C Amine (6.31 g, 35.43 mmol, 1 equiv), reacted at 25 °C for 1 hour. After completion of the reaction, dilute with water (200 ml), adjust the pH to 8 with saturated aqueous sodium bicarbonate, extract with ethyl acetate (200 ml × 2), and wash the combined organic phase with saturated aqueous salt solution (200 ml). After drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-10:1) to obtain compound 55-2.
1H NMR(400MHz,CD 3OD)δ＝6.97(dd,J＝7.2,8.7Hz,1H),6.48(dd,J＝1.8,8.7Hz,1H),3.85(d,J＝1.0Hz,3H). 1 H NMR (400MHz, CD 3 OD) δ = 6.97 (dd, J = 7.2, 8.7Hz, 1H), 6.48 (dd, J = 1.8, 8.7Hz, 1H), 3.85 (d, J = 1.0Hz, 3H ).
第二步second step
将化合物55-2(4克，18.18毫摩尔，1当量)，化合物E(5.75克，18.18毫摩尔，1当量)溶于二氧六环(40毫升)，接着向反应液中加入甲烷磺酸(1.75克，18.18毫摩尔，1.29毫升，1当量)，氮气置换三次，在100摄氏度下反应12小时。反应完成后，加水(20毫升)稀释，然后用4摩尔每升的氢氧化钠水溶液调节pH值至8，乙酸乙酯萃取(20毫升×2)，合并的有机相用饱和食盐水溶液(20毫升)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩得到的残渣用乙酸乙酯(40毫升)在25摄氏度下打浆30分钟，得到化合物55-3。Compound 55-2 (4 g, 18.18 mmol, 1 eq), compound E (5.75 g, 18.18 mmol, 1 eq) were dissolved in dioxane (40 ml), then methanesulfonic acid was added to the reaction solution (1.75 g, 18.18 mmol, 1.29 ml, 1 equiv), replaced with nitrogen three times, and reacted at 100 degrees Celsius for 12 hours. After the completion of the reaction, add water (20 ml) to dilute, then adjust the pH value to 8 with 4 moles of aqueous sodium hydroxide solution per liter, extract with ethyl acetate (20 ml × 2), and combine the organic phase with saturated saline solution (20 ml ), the organic phase was dried with anhydrous sodium sulfate, filtered, and the residue obtained by concentration was slurried with ethyl acetate (40 ml) at 25 degrees Celsius for 30 minutes to obtain compound 55-3.
MS-ESI计算值[M+H] +501，实测值501。 MS-ESI calculated [M+H] + 501, found 501.
1H NMR(400MHz,CD 3OD)δ＝8.20(dd,J＝4.3,8.3Hz,1H),8.11(s,1H),7.79(dd,J＝1.8,9.1Hz,1H),7.70-7.51(m,2H),7.37-7.25(m,1H),7.04(dd,J＝7.3,9.0Hz,1H),3.93(d,J＝1.6Hz,3H),1.83(d,J＝13.4Hz,6H). 1 H NMR (400MHz, CD 3 OD) δ = 8.20 (dd, J = 4.3, 8.3Hz, 1H), 8.11 (s, 1H), 7.79 (dd, J = 1.8, 9.1Hz, 1H), 7.70-7.51 (m,2H),7.37-7.25(m,1H),7.04(dd,J=7.3,9.0Hz,1H),3.93(d,J=1.6Hz,3H),1.83(d,J=13.4Hz, 6H).
第三步third step
将化合物55-3(300毫克，600.36微摩尔，1当量)，化合物A(108.07毫克，660.4微摩尔，1.1当量，盐酸盐)溶于二氧六环(5毫升)，接着向反应液中加入叔丁醇钠(201.93毫克，2.1毫摩尔，3.5当量)，(±)-2，2-双(二苯膦基)-1，1-联萘(74.77毫克，120.07微摩尔，0.2当量)，醋酸钯(13.48毫克，60.04微摩尔，0.1当量)，氮气置换三次，在氮气保护和110摄氏度下反应12小时。反应完成后，加水(20毫升)，乙酸乙酯萃取(20毫升×2)，合并的有机相用饱和食盐水溶液(20毫升)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩的残渣经高效液相色谱(色谱柱：Phenomenex Luna C18 150毫米×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：38％-68％，9分钟)分离得到化合物55。Compound 55-3 (300 mg, 600.36 micromoles, 1 equivalent), compound A (108.07 mg, 660.4 micromoles, 1.1 equivalents, hydrochloride) were dissolved in dioxane (5 ml), and then added to the reaction solution Add sodium tert-butoxide (201.93 mg, 2.1 mmol, 3.5 equiv), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (74.77 mg, 120.07 micromol, 0.2 equiv) , palladium acetate (13.48 mg, 60.04 micromole, 0.1 eq), replaced by nitrogen three times, and reacted for 12 hours under nitrogen protection at 110 degrees Celsius. After the reaction was complete, add water (20 ml), extract with ethyl acetate (20 ml × 2), wash the combined organic phase with saturated brine solution (20 ml), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate the residue through High performance liquid chromatography (chromatographic column: Phenomenex Luna C18 150 millimeters * 25 millimeters * 10 microns; Mobile phase: mobile phase A: formic acid aqueous solution; Mobile phase B: acetonitrile; B%: 38%-68%, 9 minutes) separates and obtains Compound 55.
MS-ESI计算值[M+H] +546，实测值546。 MS-ESI calculated [M+H] + 546, found 546.
1H NMR(400MHz,CD 3OD)δ＝8.33(dd,J＝4.2,8.3Hz,1H),8.06(br s,1H),7.61(ddd,J＝1.3,7.8,14.1Hz,1H),7.50(br t,J＝7.9Hz,1H),7.37(dd,J＝1.5,8.9Hz,1H),7.26(dt,J＝1.3,7.5Hz,1H),6.19(t,J＝9.1Hz,1H),3.97-3.81(m,8H),3.26(s,3H),2.64-2.49(m,2H),2.21-2.07(m,2H),1.85(d,J＝13.4Hz,6H). 1 H NMR (400MHz, CD 3 OD) δ = 8.33 (dd, J = 4.2, 8.3Hz, 1H), 8.06 (br s, 1H), 7.61 (ddd, J = 1.3, 7.8, 14.1Hz, 1H), 7.50(br t, J=7.9Hz, 1H), 7.37(dd, J=1.5, 8.9Hz, 1H), 7.26(dt, J=1.3, 7.5Hz, 1H), 6.19(t, J=9.1Hz, 1H),3.97-3.81(m,8H),3.26(s,3H),2.64-2.49(m,2H),2.21-2.07(m,2H),1.85(d,J＝13.4Hz,6H).
实施例56Example 56
Figure PCTCN2022129958-appb-000103
Figure PCTCN2022129958-appb-000103
第一步first step
将化合物28-2(658.58克，2.99毫摩尔，1当量)，化合物51-3(658.58毫克，2.99毫摩尔，1当量)溶于二氧六环(10毫升)，接着向反应液中加入甲烷磺酸(287.66毫克，2.99毫摩尔，213.08微升，1当量)，氮气置换三次，在100摄氏度下反应12小时。反应完成后，加水(20毫升)，然后用4摩尔每升的氢氧化钠水溶液调节pH值至8，乙酸乙酯萃取(20毫升×2)，合并的有机相用饱和食盐水溶液(20毫升)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩得到的残渣用乙酸乙酯(8毫升)在25摄氏度下打浆30分钟，得到化合物56-1。Compound 28-2 (658.58 g, 2.99 mmol, 1 eq), compound 51-3 (658.58 mg, 2.99 mmol, 1 eq) were dissolved in dioxane (10 ml), then methane was added to the reaction solution Sulfonic acid (287.66 mg, 2.99 mmol, 213.08 microliters, 1 eq.) was replaced with nitrogen three times, and reacted at 100 degrees Celsius for 12 hours. After the reaction was complete, add water (20 ml), then adjust the pH value to 8 with 4 moles of aqueous sodium hydroxide solution per liter, extract with ethyl acetate (20 ml × 2), and combine the organic phases with saturated saline solution (20 ml) After washing, the organic phase was dried over anhydrous sodium sulfate, filtered, and the concentrated residue was slurried with ethyl acetate (8 mL) at 25°C for 30 minutes to obtain compound 56-1.
MS-ESI计算值[M+H] +519，实测值519。 MS-ESI calculated [M+H] + 519, found 519.
第二步second step
将化合物56-1(500毫克，965.83微摩尔，1当量)，化合物A(173.86毫克，1.06毫摩尔，1.1当量，盐酸盐)溶于二氧六环(10毫升)，接着向反应液中加入叔丁醇钠(324.87毫克，3.38毫摩尔，3.5当量)，(±)-2，2-双(二苯膦基)-1，1-联萘(120.28毫克，193.17微摩尔，0.2当量)，醋酸钯(21.68毫克，96.58微摩尔，0.1当量)，氮气置换三次，在110摄氏度下反应12小时。反应完成后，将反应液过滤，再加水(20毫升)稀释，乙酸乙酯萃取(20毫升×2)，合并的有机相用饱和食盐水溶液(20毫升)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩的残渣经高效液相色谱(色谱柱：YMC Triart C18 150毫米×25毫米×5微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：59％-89％，8.5分钟)分离得到化合物56。Compound 56-1 (500 mg, 965.83 μmol, 1 eq), compound A (173.86 mg, 1.06 mmol, 1.1 eq, hydrochloride) were dissolved in dioxane (10 ml), and then added to the reaction solution Add sodium tert-butoxide (324.87 mg, 3.38 mmol, 3.5 equiv), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (120.28 mg, 193.17 micromol, 0.2 equiv) , palladium acetate (21.68 mg, 96.58 micromole, 0.1 eq), nitrogen replacement three times, reacted at 110 degrees Celsius for 12 hours. After the reaction was completed, the reaction solution was filtered, diluted with water (20 ml), extracted with ethyl acetate (20 ml × 2), the combined organic phase was washed with saturated saline solution (20 ml), and the organic phase was dried over anhydrous sodium sulfate. , filtered, and the concentrated residue was subjected to high performance liquid chromatography (chromatographic column: YMC Triart C18 150 mm × 25 mm × 5 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 59%-89 %, 8.5 min) to isolate compound 56.
MS-ESI计算值[M+H] +564，实测值564。 MS-ESI calculated [M+H] + 564, found 564.
1H NMR(400MHz,CD 3OD)δ＝8.28(td,J＝3.0,11.9Hz,1H),8.16(s,1H),7.74-7.49(m,1H),7.25(dd,J＝3.0,10.8Hz,1H),7.09-6.82(m,1H),5.93(dd,J＝2.9,10.4Hz,1H),3.95-3.72(m,5H),3.63(s,3H),3.23(s,3H),2.57-2.45(m,2H),2.18-2.01(m,2H),1.86(d,J＝13.6Hz,6H). 1 H NMR (400MHz, CD 3 OD) δ = 8.28 (td, J = 3.0, 11.9 Hz, 1H), 8.16 (s, 1H), 7.74-7.49 (m, 1H), 7.25 (dd, J = 3.0, 10.8Hz, 1H), 7.09-6.82(m, 1H), 5.93(dd, J=2.9, 10.4Hz, 1H), 3.95-3.72(m, 5H), 3.63(s, 3H), 3.23(s, 3H ),2.57-2.45(m,2H),2.18-2.01(m,2H),1.86(d,J＝13.6Hz,6H).
实施例57Example 57
Figure PCTCN2022129958-appb-000104
Figure PCTCN2022129958-appb-000104
第一步first step
将化合物57-1(0.9克，6.03毫摩尔，1当量)溶于二氯甲烷(3毫升)，接着在-10摄氏度下分批次向反应液中加入N-溴代丁二酰亚胺(966.31毫克，5.43毫摩尔，0.9当量)，在0摄氏度下反应1小时。反应完成后，在0摄氏度下加水(10毫升)稀释，二氯甲烷萃取(10毫升×3)，合并的有机相用饱和食盐水溶液(20毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝1：0-100：1)纯化得到得到化合物57-2。Compound 57-1 (0.9 g, 6.03 mmol, 1 equiv) was dissolved in dichloromethane (3 ml), and N-bromosuccinimide ( 966.31 mg, 5.43 mmol, 0.9 eq), reacted at 0°C for 1 hour. After the reaction was completed, dilute with water (10 ml) at 0°C, extract with dichloromethane (10 ml × 3), wash the combined organic phase with saturated saline solution (20 ml × 2), and dry the organic phase with anhydrous sodium sulfate , filtered, and the residue concentrated under reduced pressure was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:0-100:1) to obtain compound 57-2.
MS-ESI计算值[M+H] +230，实测值230。 MS-ESI calculated [M+H] + 230, found 230.
1HNMR(400MHz,DMSO-d 6)δ＝6.82(d,J＝8.4Hz,1H),6.46-6.39(m,1H),4.76(br s,2H),4.21-4.01(m,2H),2.07-1.74(m,2H). 1 HNMR (400MHz, DMSO-d 6 )δ=6.82(d,J=8.4Hz,1H),6.46-6.39(m,1H),4.76(br s,2H),4.21-4.01(m,2H), 2.07-1.74(m,2H).
第二步second step
将化合物57-2(0.68克，2.98毫摩尔，1当量)，化合物E(848.22毫克，2.68毫摩尔，0.9当量)溶于二氧六环(10毫升)，接着向反应液中加入甲烷磺酸(286.54毫克，2.98毫摩尔，1当量)，在100摄氏度下反应12小时。反应完成后，加水(30毫升)，乙酸乙酯(30毫升)稀释，然后用2摩尔每升的氢氧化钠 水溶液调节pH值至7-8，乙酸乙酯萃取(30毫升×3)，合并的有机相用饱和食盐水溶液(50毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩得到化合物57-3。Compound 57-2 (0.68 g, 2.98 mmol, 1 eq), compound E (848.22 mg, 2.68 mmol, 0.9 eq) were dissolved in dioxane (10 ml), then methanesulfonic acid was added to the reaction solution (286.54 mg, 2.98 mmol, 1 equiv), reacted at 100 degrees Celsius for 12 hours. After completion of the reaction, add water (30 ml), dilute with ethyl acetate (30 ml), then adjust the pH value to 7-8 with 2 moles of aqueous sodium hydroxide solution per liter, extract with ethyl acetate (30 ml×3), and combine The organic phase was washed with saturated saline solution (50 ml×2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 57-3.
MS-ESI计算值[M+H] +509，实测值509。 MS-ESI calculated [M+H] +509 , found 509.
第三步third step
将化合物57-3(0.8克，1.58毫摩尔，1当量)，化合物A(283.62毫克，1.73毫摩尔，1.1当量)溶于二氧六环(12毫升)，接着向反应液中加入叔丁醇钠(454.24毫克，4.73毫摩尔，3当量)，(±)-2，2-双(二苯膦基)-1，1-联萘(196.21毫克，315.12微摩尔，0.2当量)，醋酸钯(35.37毫克，157.56微摩尔，0.1当量)，置换三次氮气，然后在110摄氏度和氮气保护下反应16小时。反应完成后，反应液浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝1：1-0：1)纯化的粗品经高效液相色谱(色谱柱：Phenomenex luna C18 150毫米×40毫米×15微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：20％-50％，15分钟)分离得到化合物57。Compound 57-3 (0.8 g, 1.58 mmol, 1 eq), compound A (283.62 mg, 1.73 mmol, 1.1 eq) were dissolved in dioxane (12 ml), then tert-butanol was added to the reaction solution Sodium (454.24 mg, 4.73 mmol, 3 equivalents), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (196.21 mg, 315.12 micromoles, 0.2 equivalents), palladium acetate ( 35.37 mg, 157.56 micromole, 0.1 equivalent), nitrogen replacement three times, and then reacted at 110 degrees Celsius under nitrogen protection for 16 hours. After the reaction was completed, the concentrated residue of the reaction solution was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:1-0:1) and the crude product was purified by high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 mm × 40 mm × 15 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 20%-50%, 15 minutes) isolated and obtained compound 57.
MS-ESI计算值[M+H] +554，实测值554。 MS-ESI calculated [M+H] +554 , found 554.
1HNMR(400MHz,DMSO-d 6)δ＝11.16(s,1H),8.50(br d,J＝3.4Hz,1H),8.06(s,1H),7.90(s,1H),7.61-7.49(m,1H),7.31(br t,J＝9.0Hz,2H),7.12(br t,J＝7.3Hz,1H),5.99(d,J＝8.6Hz,1H),4.11-4.06(m,2H),3.83-3.68(m,5H),3.17(s,2H),3.13(s,3H),2.47-2.42(m,2H),2.09-1.99(m,2H),1.87-1.80(m,2H),1.77(d,J＝13.6Hz,6H). 1 H NMR (400MHz, DMSO-d 6 ) δ=11.16(s,1H), 8.50(br d,J=3.4Hz,1H), 8.06(s,1H),7.90(s,1H),7.61-7.49( m,1H),7.31(br t,J=9.0Hz,2H),7.12(br t,J=7.3Hz,1H),5.99(d,J=8.6Hz,1H),4.11-4.06(m,2H ),3.83-3.68(m,5H),3.17(s,2H),3.13(s,3H),2.47-2.42(m,2H),2.09-1.99(m,2H),1.87-1.80(m,2H ), 1.77(d,J=13.6Hz,6H).
实施例58Example 58
Figure PCTCN2022129958-appb-000105
Figure PCTCN2022129958-appb-000105
第一步first step
将化合物26-1(2.00克，4.15毫摩尔，1当量)，化合物58-1(2.00克，6.23毫摩尔，1.5当量)，氯(2-二环己基膦基-2，4，6-三异丙基-1，1-联苯基)[2-(2-氨基-1，1-联苯)]钯(II)(326.67毫克，415.19微摩尔，0.1当量)，2-二环己基磷-2，4，6-三异丙基联苯(395.85毫克，830.38微摩尔，0.2当量)溶于二氧六环(20毫升)，置换三次氮气，并在氮气氛围和90摄氏度下反应12小时。反应完成后，向反应液中加入饱和氟化钾水溶液(20毫升)，乙酸乙酯(20毫升×2)萃取，饱和食盐水(20毫升×2)洗涤。有机相经无水硫酸钠干燥后，过滤浓缩得到粗品。粗品经硅胶柱层析(乙酸乙酯：甲醇＝100：1-10：1)分离纯化得到化合物58-2。Compound 26-1 (2.00 g, 4.15 mmol, 1 eq), compound 58-1 (2.00 g, 6.23 mmol, 1.5 eq), chloro(2-dicyclohexylphosphino-2,4,6-tri Isopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II) (326.67 mg, 415.19 micromoles, 0.1 equiv), 2-dicyclohexylphosphine -2,4,6-Triisopropylbiphenyl (395.85 mg, 830.38 micromole, 0.2 eq) was dissolved in dioxane (20 ml), nitrogen was replaced three times, and reacted under nitrogen atmosphere at 90 degrees Celsius for 12 hours . After the reaction was completed, saturated potassium fluoride aqueous solution (20 mL) was added to the reaction liquid, extracted with ethyl acetate (20 mL×2), and washed with saturated brine (20 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (ethyl acetate:methanol=100:1-10:1) to obtain compound 58-2.
MS-ESI计算值[M+H] +433,实测值433。 MS-ESI calculated [M+H] + 433, found 433.
第二步second step
将化合物58-2(500毫克，1.16毫摩尔，1当量)溶于二氯甲烷(10毫升)，然后加入戴斯-马丁过碘烷(734.93毫克，1.73毫摩尔，1.5当量)，并在25摄氏度下反应0.5小时。反应完成后，向反应液中加入饱和碳酸氢钠水溶液(20毫升)和饱和亚硫酸钠水溶液(20毫升)，再用二氯甲烷(20毫升×2)萃取，饱和食盐水(20毫升×2)洗涤。有机相经无水硫酸钠干燥后，过滤浓缩得到化合物58-3。Compound 58-2 (500 mg, 1.16 mmol, 1 eq) was dissolved in dichloromethane (10 ml), then Dess-Martin periodinane (734.93 mg, 1.73 mmol, 1.5 eq) was added, and the The reaction was carried out at 0.5°C for 0.5 hours. After the reaction was completed, saturated aqueous sodium bicarbonate (20 ml) and saturated aqueous sodium sulfite (20 ml) were added to the reaction solution, extracted with dichloromethane (20 ml × 2), washed with saturated brine (20 ml × 2) . The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 58-3.
第三步third step
将化合物58-3(200毫克，464.23微摩尔，1当量)，化合物A(113.95毫克，696.34微摩尔，1.5当量)溶于二氯甲烷(10毫升)，然后加入N，N-二异丙基乙胺(60.00毫克，464.23微摩尔，80.86微升，1当量)和三乙酰氧基硼氢化钠(196.78毫克，928.45微摩尔，2当量)，并在25摄氏度下反应0.5小时。反应完成后，反应液经过滤浓缩得到粗品。粗品经高效液相色谱(色谱柱：Phenomenex luna C18 150毫米×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:12％-32％，9分钟)分离得到化合物58(甲酸盐)。Compound 58-3 (200 mg, 464.23 micromoles, 1 equivalent), compound A (113.95 mg, 696.34 micromoles, 1.5 equivalents) were dissolved in dichloromethane (10 ml), and N,N-diisopropyl Ethylamine (60.00 mg, 464.23 micromoles, 80.86 microliters, 1 equivalent) and sodium triacetoxyborohydride (196.78 mg, 928.45 micromoles, 2 equivalents), and reacted at 25 degrees Celsius for 0.5 hours. After the reaction was completed, the reaction solution was filtered and concentrated to obtain a crude product. The crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 mm × 25 mm × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 12%-32%, 9 minutes) Compound 58 (formate salt) was isolated.
MS-ESI计算值[M+H] +542，实测值542。 MS-ESI calculated [M+H] + 542, found 542.
1H NMR(400MHz,CD 3OD)δ＝8.47(s,1H),8.33-8.26(m,1H),8.14(s,1H),8.04-7.94(m,1H),7.69-7.58(m,1H),7.54(t,J＝7.9Hz,1H),7.34-7.24(m,1H),7.09(d,J＝5.3Hz,2H),4.31(s,2H),4.05(br d,J＝17.1Hz,4H),3.83-3.75(m,4H),3.20(s,3H),2.60-2.50(m,2H),2.19-2.07(m,2H),1.86(s,3H),1.83(s,3H)。 1 H NMR (400MHz, CD 3 OD) δ=8.47(s,1H),8.33-8.26(m,1H),8.14(s,1H),8.04-7.94(m,1H),7.69-7.58(m, 1H), 7.54(t, J=7.9Hz, 1H), 7.34-7.24(m, 1H), 7.09(d, J=5.3Hz, 2H), 4.31(s, 2H), 4.05(br d, J= 17.1Hz, 4H), 3.83-3.75(m, 4H), 3.20(s, 3H), 2.60-2.50(m, 2H), 2.19-2.07(m, 2H), 1.86(s, 3H), 1.83(s ,3H).
实施例59Example 59
Figure PCTCN2022129958-appb-000106
Figure PCTCN2022129958-appb-000106
第一步first step
将化合物59-1(2克，8.62毫摩尔，1当量)，碘甲烷(1.59克，11.21毫摩尔，697.58微升，1.3当量)和碳酸钾(2.38克，17.24毫摩尔，2当量)溶于N，N-二甲基甲酰胺(20毫升)，置换三次氮气，在60摄氏度下反应12小时。反应完成后，在25摄氏度下，向反应液中加入水(100毫升)淬灭反应，然后用乙酸乙酯(100毫升×2)萃取，合并有机相经饱和食盐水(50毫升×2)洗涤，有机相经无水硫酸钠干燥，过滤，减压浓缩，得到粗品化合物59-2。Compound 59-1 (2 g, 8.62 mmol, 1 eq), iodomethane (1.59 g, 11.21 mmol, 697.58 μl, 1.3 eq) and potassium carbonate (2.38 g, 17.24 mmol, 2 eq) were dissolved in N, N-dimethylformamide (20 ml) was replaced with nitrogen three times, and reacted at 60° C. for 12 hours. After the reaction was completed, at 25 degrees Celsius, water (100 ml) was added to the reaction solution to quench the reaction, then extracted with ethyl acetate (100 ml × 2), and the combined organic phases were washed with saturated brine (50 ml × 2) , the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude compound 59-2.
1H NMR(400MHz,CD 3OD)δ＝7.75(dd,J＝0.6,2.0Hz,1H),7.69-7.57(m,1H),3.96(s,3H),2.39(s,3H)。 1 H NMR (400 MHz, CD 3 OD) δ=7.75 (dd, J=0.6, 2.0 Hz, 1H), 7.69-7.57 (m, 1H), 3.96 (s, 3H), 2.39 (s, 3H).
第二步second step
将化合物59-2(2克，8.13毫摩尔，1当量)溶于乙醇(12毫升)和水(6毫升)中，接着向反应 液中加入铁粉(2.72克，48.77毫摩尔，6当量)和氯化铵(3.48克，65.03毫摩尔，8当量)，置换三次氮气，在70摄氏度下反应12小时。反应完成后，将反应液过滤，减压浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1到5：1)分离得到化合物59-3。Compound 59-2 (2 g, 8.13 mmol, 1 eq) was dissolved in ethanol (12 ml) and water (6 ml), then iron powder (2.72 g, 48.77 mmol, 6 eq) was added to the reaction solution and ammonium chloride (3.48 g, 65.03 mmol, 8 equivalents), nitrogen was replaced three times, and reacted at 70 degrees Celsius for 12 hours. After the reaction was completed, the reaction solution was filtered and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate=100:1 to 5:1) to obtain compound 59-3.
MS-ESI计算值[M+H] +216，实测值216。 MS-ESI calculated [M+H] + 216, found 216.
1H NMR(400MHz,CD 3OD)δ＝6.75-6.58(m,1H),6.52-6.36(m,1H),3.73(s,3H),3.47-2.75(m,2H),2.12(s,3H)。 1 H NMR (400MHz, CD 3 OD) δ=6.75-6.58(m,1H),6.52-6.36(m,1H),3.73(s,3H),3.47-2.75(m,2H),2.12(s, 3H).
第三步third step
将化合物59-3(1克，4.63毫摩尔，1当量)，化合物E(1.76克，5.55毫摩尔，1.2当量)和甲基磺酸(444.78毫克，4.63毫摩尔，329.47微升，1当量)溶于二氧六环(10毫升)，置换三次氮气，在100摄氏度下反应12小时。反应完成后，在25摄氏度下，向反应液中加水(10毫升)和乙酸乙酯(10毫升)，并用氢氧化钠水溶液(4摩尔每升)调节pH至8，然后用乙酸乙酯(10毫升×2)萃取，合并有机相经饱和食盐水(10毫升×2)洗涤，有机相经无水硫酸钠干燥，过滤，减压浓缩。粗品经乙酸乙酯(10毫升)打浆，得到化合物59-4。Compound 59-3 (1 g, 4.63 mmol, 1 eq), compound E (1.76 g, 5.55 mmol, 1.2 eq) and methanesulfonic acid (444.78 mg, 4.63 mmol, 329.47 μl, 1 eq) Dissolve in dioxane (10 ml), replace nitrogen three times, and react at 100 degrees Celsius for 12 hours. After the reaction was completed, at 25 degrees Celsius, water (10 milliliters) and ethyl acetate (10 milliliters) were added to the reaction solution, and the pH was adjusted to 8 with aqueous sodium hydroxide solution (4 moles per liter), and then ethyl acetate (10 mL×2), the combined organic phases were washed with saturated brine (10 mL×2), the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was slurried with ethyl acetate (10 mL) to give compound 59-4.
MS-ESI计算值[M+H] +497，实测值497。 1H NMR(400MHz,CD 3OD)δ＝8.27(dd,J＝4.4,8.1Hz,1H),8.15(s,1H),7.80(d,J＝1.4Hz,1H),7.73-7.53(m,2H),7.40-7.26(m,1H),7.04(d,J＝1.3Hz,1H),3.81(s,3H),2.15(s,3H),1.87(d,J＝13.5Hz,6H)。 MS-ESI calculated [M+H] + 497, found 497. 1 H NMR (400MHz, CD 3 OD) δ=8.27(dd, J=4.4, 8.1Hz, 1H), 8.15(s, 1H), 7.80(d, J=1.4Hz, 1H), 7.73-7.53(m ,2H),7.40-7.26(m,1H),7.04(d,J=1.3Hz,1H),3.81(s,3H),2.15(s,3H),1.87(d,J=13.5Hz,6H) .
第四步the fourth step
将化合物59-4(2克，4.03毫摩尔，1当量)，化合物A(726.23毫克，4.44毫摩尔，1.1当量，盐酸盐)，叔丁醇钠(1.16克，12.10毫摩尔，3当量)，(±)-2，2-双(二苯膦基)-1，1-联萘(502.42毫克，806.88微摩尔，0.2当量)，醋酸钯(90.58毫克，403.44微摩尔，0.1当量)溶于二氧六环(20毫升)，置换三次氮气，并在氮气氛围下在110摄氏度下反应12小时。反应完成后，反应液过滤，滤液浓缩。粗品经硅胶柱层析(石油醚：乙酸乙酯＝100：1到乙酸乙酯：甲醇＝50：1)，纯化得到的粗品经高效液相色谱(色谱柱：Phenomenex Luna C18 150毫米×40毫米×15微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：37％-67％，15分钟)分离得到化合物59。Compound 59-4 (2 g, 4.03 mmol, 1 eq), compound A (726.23 mg, 4.44 mmol, 1.1 eq, hydrochloride), sodium tert-butoxide (1.16 g, 12.10 mmol, 3 eq) , (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (502.42 mg, 806.88 micromole, 0.2 eq), palladium acetate (90.58 mg, 403.44 micromole, 0.1 eq) was dissolved in Dioxane (20 ml) was replaced with nitrogen three times, and reacted at 110° C. for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate=100:1 to ethyl acetate:methanol=50:1), and the purified crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Luna C18 150 mm×40 mm × 15 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 37%-67%, 15 minutes) isolated and obtained compound 59.
MS-ESI计算值[M+H] +542,实测值542. MS-ESI calculated value [M+H] + 542, measured value 542.
1H NMR(400MHz,CD 3OD)δ＝8.31(dd,J＝4.5,8.0Hz,1H),8.09(s,1H),7.64(ddd,J＝1.3,7.7,13.9Hz,1H),7.54(t,J＝7.9Hz,1H),7.35-7.19(m,2H),6.01(d,J＝1.5Hz,1H),3.88-3.76(m,5H),3.59-3.59(m,1H),3.61(s,2H),3.23(s,3H),2.50(ddd,J＝2.8,6.8,9.8Hz,2H),2.13-2.04(m,5H),1.84(d,J＝13.4Hz,6H)。 1 H NMR (400MHz, CD 3 OD) δ = 8.31 (dd, J = 4.5, 8.0Hz, 1H), 8.09 (s, 1H), 7.64 (ddd, J = 1.3, 7.7, 13.9Hz, 1H), 7.54 (t,J=7.9Hz,1H),7.35-7.19(m,2H),6.01(d,J=1.5Hz,1H),3.88-3.76(m,5H),3.59-3.59(m,1H), 3.61(s,2H),3.23(s,3H),2.50(ddd,J=2.8,6.8,9.8Hz,2H),2.13-2.04(m,5H),1.84(d,J=13.4Hz,6H) .
实施例60Example 60
Figure PCTCN2022129958-appb-000107
Figure PCTCN2022129958-appb-000107
第一步first step
将化合物28-3(500毫克，1.00毫摩尔，1当量)，化合物58-1(481.92毫克，1.50毫摩尔，1.5当量)，氯(2-二环己基膦基-2，4，6-三异丙基-1，1-联苯基)[2-(2-氨基-1，1-联苯)]钯(II)(157.45毫克，200.12微摩尔，0.2当量)溶于二氧六环(10毫升)，置换三次氮气，并在氮气氛围下在80摄氏度下反应12小时。反应完成后，向反应液中加入饱和氟化钾水溶液(20毫升)，乙酸乙酯(20毫升×2)萃取，饱和食盐水(20毫升×2)洗涤。有机相经无水硫酸钠干燥后，过滤浓缩得到粗品。粗品经硅胶柱层析(乙酸乙酯：甲醇＝100：1-10：1)分离纯化得到化合物60-1。Compound 28-3 (500 mg, 1.00 mmol, 1 equivalent), compound 58-1 (481.92 mg, 1.50 mmol, 1.5 equivalents), chloro(2-dicyclohexylphosphino-2,4,6-tri Isopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II) (157.45 mg, 200.12 micromole, 0.2 equiv) was dissolved in dioxane ( 10 ml), nitrogen was replaced three times, and reacted at 80 degrees Celsius for 12 hours under a nitrogen atmosphere. After the reaction was completed, saturated potassium fluoride aqueous solution (20 mL) was added to the reaction liquid, extracted with ethyl acetate (20 mL×2), and washed with saturated brine (20 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was separated and purified by silica gel column chromatography (ethyl acetate:methanol=100:1-10:1) to obtain compound 60-1.
MS-ESI计算值[M+H] +451,实测值451。 MS-ESI calculated [M+H] + 451, found 451.
第二步second step
将化合物60-1(150毫克，332.72微摩尔，1当量)溶于二氯甲烷(5毫升)，然后加入戴斯-马丁过碘烷(211.68毫克，499.08毫摩尔，1.5当量)，并在25摄氏度下反应0.5小时。反应完成后，向反应液中加入饱和碳酸氢钠水溶液(20毫升)和饱和亚硫酸钠水溶液(20毫升)，再用二氯甲烷(20毫升×2)萃取，饱和食盐水(20毫升×2)洗涤。有机相经无水硫酸钠干燥后，过滤浓缩得到化合物60-2。Compound 60-1 (150 mg, 332.72 micromole, 1 eq) was dissolved in dichloromethane (5 ml), then Dess-Martin periodinane (211.68 mg, 499.08 mmol, 1.5 eq) was added, and the The reaction was carried out at 0.5°C for 0.5 hours. After the reaction was completed, saturated aqueous sodium bicarbonate (20 ml) and saturated aqueous sodium sulfite (20 ml) were added to the reaction solution, extracted with dichloromethane (20 ml × 2), washed with saturated brine (20 ml × 2) . The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 60-2.
第三步third step
将化合物60-2(100毫克，222.81微摩尔，1当量)，化合物A(54.69毫克，334.21微摩尔，1.5当量)溶于二氯甲烷(5毫升)，然后加入N，N-二异丙基乙胺(28.80毫克，222.81微摩尔，38.81微升，1当量)和三乙酰氧基硼氢化钠(94.44毫克，445.62微摩尔，2当量)，并在25摄氏度下反应0.5小时。反应完成后，反应液经过滤浓缩得到粗品。粗品经高效液相色谱(色谱柱：Phenomenex luna C18 150毫米×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:10％-40％)分离得到化合物60。Compound 60-2 (100 mg, 222.81 micromoles, 1 equivalent), compound A (54.69 mg, 334.21 micromoles, 1.5 equivalents) were dissolved in dichloromethane (5 ml), and N, N-diisopropyl Ethylamine (28.80 mg, 222.81 micromoles, 38.81 microliters, 1 equiv) and sodium triacetoxyborohydride (94.44 mg, 445.62 micromoles, 2 equivs), and reacted at 25 degrees Celsius for 0.5 hours. After the reaction was completed, the reaction solution was filtered and concentrated to obtain a crude product. The crude product was separated by high performance liquid chromatography (column: Phenomenex luna C18 150 mm × 25 mm × 10 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 10%-40%) to obtain the compound 60.
MS-ESI计算值[M+H] +560，实测值560。 MS-ESI calculated [M+H] + 560, found 560.
1H NMR(400MHz,CD 3OD)δ＝8.47(s,1H),8.33-8.26(m,1H),8.14(s,1H),8.04-7.94(m,1H),7.69-7.58(m,1H),7.54(t,J＝7.9Hz,1H),7.34-7.24(m,1H),7.09(d,J＝5.3Hz,2H),4.31(s,2H),4.05(br d,J＝17.1Hz,4H),3.83-3.75(m,4H),3.20(s,3H),2.60-2.50(m,2H),2.19-2.07(m,2H),1.86(s,3H),1.83(s,3H)。 1 H NMR (400MHz, CD 3 OD) δ=8.47(s,1H),8.33-8.26(m,1H),8.14(s,1H),8.04-7.94(m,1H),7.69-7.58(m, 1H), 7.54(t, J=7.9Hz, 1H), 7.34-7.24(m, 1H), 7.09(d, J=5.3Hz, 2H), 4.31(s, 2H), 4.05(br d, J= 17.1Hz, 4H), 3.83-3.75(m, 4H), 3.20(s, 3H), 2.60-2.50(m, 2H), 2.19-2.07(m, 2H), 1.86(s, 3H), 1.83(s ,3H).
实施例61Example 61
Figure PCTCN2022129958-appb-000108
Figure PCTCN2022129958-appb-000108
第一步first step
将化合物61-1(5克，33.08毫摩尔，1当量)溶于乙腈(60毫升)，接着在0摄氏度下向反应液中加入N-溴代丁二酰亚胺(6.2克，34.7毫摩尔，1.05当量)，在30摄氏度下反应1.5小时。反应完成后，加水(100毫升)稀释，乙酸乙酯萃取(50毫升×3)，合并的有机相用饱和氯化钠水溶液(80毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-20：1)纯化的粗品经高效液相色谱(色谱柱：Phenomenex luna C18 150毫米×40毫米×15微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:20％-50％，15分钟)分离得到得到化合物61-2。Compound 61-1 (5 g, 33.08 mmol, 1 equivalent) was dissolved in acetonitrile (60 ml), and then N-bromosuccinimide (6.2 g, 34.7 mmol) was added to the reaction solution at 0° C. , 1.05 equivalents), reacted at 30 degrees Celsius for 1.5 hours. After completion of the reaction, dilute with water (100 ml), extract with ethyl acetate (50 ml × 3), wash the combined organic phase with saturated aqueous sodium chloride solution (80 ml × 2), dry the organic phase with anhydrous sodium sulfate, filter , the residue concentrated under reduced pressure is purified by silica gel column chromatography (petroleum ether: ethyl acetate=100:1-20:1) and the crude product is purified by high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 mm × 40 mm × 15 microns ; Mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 20%-50%, 15 minutes) separated to obtain compound 61-2.
MS-ESI计算值[M+H] +230，实测值230。 MS-ESI calculated [M+H] + 230, found 230.
1HNMR(400MHz,DMSO-d 6)δ＝6.77(d,J＝8.6Hz,1H),6.20(d,J＝8.7Hz,1H),4.89(s,2H),4.35-4.17(m,4H). 1 HNMR (400MHz, DMSO-d 6 )δ=6.77(d, J=8.6Hz, 1H), 6.20(d, J=8.7Hz, 1H), 4.89(s, 2H), 4.35-4.17(m, 4H ).
第二步second step
将化合物61-2(1.28克，5.56毫摩尔，1当量)，化合物E(1.58克，5.01毫摩尔，0.9当量)溶于二氧六环(20毫升)，接着向反应液中加入甲烷磺酸(534.71毫克，5.56毫摩尔，1当量)，在100摄氏度下反应12小时。反应完成后，加水(30毫升)，乙酸乙酯(30毫升)稀释，然后用2摩尔每升的氢氧化钠水溶液调节pH值至7-8，乙酸乙酯萃取(30毫升×3)，合并的有机相用饱和氯化钠水溶液(50毫升×2)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩得到化合物61-3。Compound 61-2 (1.28 g, 5.56 mmol, 1 eq), compound E (1.58 g, 5.01 mmol, 0.9 eq) were dissolved in dioxane (20 ml), then methanesulfonic acid was added to the reaction solution (534.71 mg, 5.56 mmol, 1 equivalent), reacted at 100 degrees Celsius for 12 hours. After completion of the reaction, add water (30 ml), dilute with ethyl acetate (30 ml), then adjust the pH value to 7-8 with 2 moles of aqueous sodium hydroxide solution per liter, extract with ethyl acetate (30 ml×3), and combine The organic phase was washed with saturated aqueous sodium chloride (50 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 61-3.
MS-ESI计算值[M+H] +511，实测值511。 MS-ESI calculated [M+H] + 511, found 511.
第三步third step
将化合物61-3(1克，1.96毫摩尔，1当量)，化合物A(353.15毫克，2.16毫摩尔，1.1当量)溶于二氧六环(20毫升)，接着向反应液中加入叔丁醇钠(565.63毫克，5.89毫摩尔，3当量)，(±)-2，2-双(二苯膦基)-1，1-联萘(244.32毫克，392.37微摩尔，0.2当量)，醋酸钯(44.05毫克，196.19微摩尔，0.1当量)，在110摄氏度下反应16小时。反应完成后，反应液浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝1：1-0：1)纯化的粗品经高效液相色谱(色谱柱：Phenomenex luna C18 150毫米×40毫米×15微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％:18％-48％，15分钟)分离得到化合物61(甲酸盐)。Compound 61-3 (1 g, 1.96 mmol, 1 eq), compound A (353.15 mg, 2.16 mmol, 1.1 eq) were dissolved in dioxane (20 ml), and then tert-butanol was added to the reaction solution Sodium (565.63 mg, 5.89 mmol, 3 equivalents), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (244.32 mg, 392.37 micromoles, 0.2 equivalents), palladium acetate ( 44.05 mg, 196.19 micromole, 0.1 equivalent), reacted at 110 degrees Celsius for 16 hours. After the reaction was completed, the concentrated residue of the reaction solution was purified by silica gel column chromatography (petroleum ether: ethyl acetate=1:1-0:1) and the crude product was purified by high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 mm × 40 mm × 15 microns; mobile phase: mobile phase A: formic acid aqueous solution; mobile phase B: acetonitrile; B%: 18%-48%, 15 minutes) isolated to obtain compound 61 (formate).
MS-ESI计算值[M+H] +556，实测值556。 MS-ESI calculated [M+H] + 556, found 556.
1HNMR(400MHz,DMSO-d 6)δ＝11.19(s,1H),8.57-8.42(m,1H),8.26-7.96(m,3H),7.61-7.45(m, 1H),7.29(br t,J＝7.4Hz,1H),7.09(t,J＝7.3Hz,1H),6.89(d,J＝8.6Hz,1H),5.92(d,J＝8.6Hz,1H),4.14(q,J＝4.8Hz,4H),3.83-3.74(m,5H),3.11(s,3H),2.43(dt,J＝3.4,6.5Hz,2H),2.05-1.94(m,2H),1.76(d,J＝13.6Hz,6H). 1 HNMR (400MHz, DMSO-d 6 )δ=11.19(s,1H),8.57-8.42(m,1H),8.26-7.96(m,3H),7.61-7.45(m,1H),7.29(br t ,J=7.4Hz,1H),7.09(t,J=7.3Hz,1H),6.89(d,J=8.6Hz,1H),5.92(d,J=8.6Hz,1H),4.14(q,J =4.8Hz, 4H), 3.83-3.74(m, 5H), 3.11(s, 3H), 2.43(dt, J=3.4, 6.5Hz, 2H), 2.05-1.94(m, 2H), 1.76(d, J=13.6Hz,6H).
实施例62Example 62
Figure PCTCN2022129958-appb-000109
Figure PCTCN2022129958-appb-000109
第一步first step
将化合物62-1(1克，7.4毫摩尔，1当量)溶于N，N-二甲基甲酰胺(10毫升)，接着在0摄氏度下向反应液中加入N-溴代丁二酰亚胺(1.32克，7.4毫摩尔，1当量)，在25摄氏度下反应1小时。反应完成后，加水(200毫升)稀释，用饱和的碳酸氢钠水溶液调pH到8，乙酸乙酯萃取(200毫升×2)，合并的有机相用饱和食盐水溶液(200毫升)洗涤，有机相用无水硫酸钠干燥，过滤，减压浓缩的残渣经硅胶柱层析(石油醚：乙酸乙酯＝100：1-20：1)纯化得到得到化合物62-2。Compound 62-1 (1 g, 7.4 mmol, 1 eq) was dissolved in N, N-dimethylformamide (10 ml), then N-bromosuccinyl was added to the reaction solution at 0°C Amine (1.32 g, 7.4 mmol, 1 equiv), reacted at 25 °C for 1 hour. After completion of the reaction, dilute with water (200 ml), adjust the pH to 8 with saturated aqueous sodium bicarbonate, extract with ethyl acetate (200 ml × 2), and wash the combined organic phase with saturated aqueous salt solution (200 ml). After drying with anhydrous sodium sulfate, filtering, and concentrating under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 100:1-20:1) to obtain compound 62-2.
MS-ESI计算值[M+H] +216，实测值216。 MS-ESI calculated [M+H] + 216, found 216.
1H NMR(400MHz,CDCl 3)δ＝6.81(d,J＝8.3Hz,1H),6.45(d,J＝8.3Hz,1H),4.62(t,J＝8.7Hz,2H),3.22(t,J＝8.7Hz,2H)。 1 H NMR (400MHz, CDCl 3 ) δ=6.81(d, J=8.3Hz, 1H), 6.45(d, J=8.3Hz, 1H), 4.62(t, J=8.7Hz, 2H), 3.22(t , J=8.7Hz, 2H).
第二步second step
将化合物62-2(1.33克，6.21毫摩尔，1当量)，化合物E(1.96克，6.21毫摩尔，1当量)溶于异丙醇(13毫升)，接着向反应液中加入甲烷磺酸(597.13毫克，6.21毫摩尔，442.32微升，1当量)，氮气置换三次，在90摄氏度下反应12小时。反应完成后，加水(20毫升)，然后用4摩尔/升的氢氧化钠水溶液调节pH至8，乙酸乙酯萃取(20毫升×2)，合并的有机相用饱和食盐水溶液(20毫升)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩得到的残渣用乙酸乙酯(40毫升)在25摄氏度下打浆30分钟，得到化合物62-3。Compound 62-2 (1.33 g, 6.21 mmol, 1 eq), compound E (1.96 g, 6.21 mmol, 1 eq) were dissolved in isopropanol (13 ml), then methanesulfonic acid ( 597.13 mg, 6.21 mmol, 442.32 microliters, 1 equivalent), nitrogen replacement three times, and reacted at 90 degrees Celsius for 12 hours. After the reaction was completed, add water (20 ml), then adjust the pH to 8 with 4 mol/L aqueous sodium hydroxide solution, extract with ethyl acetate (20 ml × 2), and wash the combined organic phase with a saturated saline solution (20 ml) , the organic phase was dried over anhydrous sodium sulfate, filtered, and the residue obtained by concentration was slurried with ethyl acetate (40 mL) at 25° C. for 30 minutes to obtain compound 62-3.
MS-ESI计算值[M+H] +495，实测值495。 MS-ESI calculated [M+H] + 495, found 495.
1H NMR(400MHz,CDCl 3)δ＝8.47(dd,J＝4.4,8.4Hz,1H),8.03(s,1H),7.86(d,J＝8.8Hz,1H),7.50-7.38(m,1H),7.25-7.17(m,1H),7.11-7.00(m,1H),6.95-6.80(m,2H),4.58(t,J＝8.8Hz,2H),3.19(t,J＝8.8Hz,2H),1.76(d,J＝13.2Hz,6H)。 1 H NMR (400MHz, CDCl 3 )δ=8.47(dd, J=4.4,8.4Hz,1H),8.03(s,1H),7.86(d,J=8.8Hz,1H),7.50-7.38(m, 1H),7.25-7.17(m,1H),7.11-7.00(m,1H),6.95-6.80(m,2H),4.58(t,J＝8.8Hz,2H),3.19(t,J＝8.8Hz , 2H), 1.76 (d, J=13.2Hz, 6H).
第三步third step
将化合物62-3(500毫克，1.01毫摩尔，1当量)，化合物A(182.3毫克，1.11毫摩尔，1.1当量，一个盐酸盐)溶于二氧六环(12毫升)，接着向反应液中加入叔丁醇钠(340.64毫克，3.54毫摩尔，3.5当量)，(±)-2，2-双(二苯膦基)-1，1-联萘(126.12毫克，202.54微摩尔，0.2当量)，醋酸钯(22.74毫克，101.27 微摩尔，0.1当量)，在110摄氏度下反应12小时。反应完成后，加水(20毫升)，乙酸乙酯萃取(20毫升×2)，合并的有机相用饱和食盐水溶液(20毫升)洗涤，有机相用无水硫酸钠干燥，过滤，浓缩的残渣经高效液相色谱(色谱柱：Phenomenex Luna C18 150毫米×25毫米×10微米；流动相：流动相A：甲酸水溶液；流动相B：乙腈；B％：23％-53％，10分钟)分离得到化合物62。Compound 62-3 (500 mg, 1.01 mmol, 1 eq), compound A (182.3 mg, 1.11 mmol, 1.1 eq, a hydrochloride salt) were dissolved in dioxane (12 ml), and then added to the reaction solution Add sodium tert-butoxide (340.64 mg, 3.54 mmol, 3.5 eq), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (126.12 mg, 202.54 micromol, 0.2 eq ), palladium acetate (22.74 mg, 101.27 micromole, 0.1 eq), reacted at 110 degrees Celsius for 12 hours. After the reaction was complete, add water (20 ml), extract with ethyl acetate (20 ml × 2), wash the combined organic phase with saturated brine solution (20 ml), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate the residue through High performance liquid chromatography (chromatographic column: Phenomenex Luna C18 150 millimeters * 25 millimeters * 10 microns; Mobile phase: mobile phase A: formic acid aqueous solution; Mobile phase B: acetonitrile; B%: 23%-53%, 10 minutes) separates and obtains Compound 62.
MS-ESI计算值[M+H] +540，实测值540。 MS-ESI calculated [M+H] + 540, found 540.
1H NMR(400MHz,DMSO-d 6)δ＝11.19(s,1H),8.53(br s,1H),8.33(s,1H),8.02(s,1H),7.65-7.44(m,1H),7.27(br d,J＝7.6Hz,1H),7.13-6.84(m,2H),5.86(d,J＝8.6Hz,1H),4.42(t,J＝8.7Hz,2H),3.97-3.72(m,5H),3.13(s,5H),2.48-2.43(m,2H),2.10-1.96(m,2H),1.76(d,J＝13.6Hz,6H)。 1 H NMR (400MHz, DMSO-d 6 )δ=11.19(s,1H),8.53(br s,1H),8.33(s,1H),8.02(s,1H),7.65-7.44(m,1H) ,7.27(br d,J=7.6Hz,1H),7.13-6.84(m,2H),5.86(d,J=8.6Hz,1H),4.42(t,J=8.7Hz,2H),3.97-3.72 (m, 5H), 3.13 (s, 5H), 2.48-2.43 (m, 2H), 2.10-1.96 (m, 2H), 1.76 (d, J=13.6Hz, 6H).
实验例1：体外评价LRRK2激酶抑制活性Experimental example 1: In vitro evaluation of LRRK2 kinase inhibitory activity
实验目的：通过均相时间分辨荧光检测磷酸化Fluorescein-ERM(LRRKtide)peptide的磷酸基团与
Figure PCTCN2022129958-appb-000110
Tb-pERM(pLRRKtide)Antibody抗体结合后产生的能量信号转移(520nM/485nM荧光信号比值)。计算待测化合物的LRRK2激酶抑制IC 50值。 The purpose of the experiment: to detect the phosphorylated Fluorescein-ERM (LRRKtide) peptide phosphate group and
Figure PCTCN2022129958-appb-000110
Energy signal transfer (520nM/485nM fluorescence signal ratio) generated after Tb-pERM (pLRRKtide) Antibody binding. Calculate the LRRK2 kinase inhibitory IC 50 value of the test compound.
实验材料:Experimental Materials:
1.反应溶液：10mM羟乙基哌嗪乙磺酸(PH7.5)；2mM氯化镁；0.5mM乙二醇二***二胺四乙酸；0.002％聚氧乙烯脂肪醇醚；1mM二硫苏糖醇和1％DMSO；1. Reaction solution: 10mM hydroxyethylpiperazineethanesulfonic acid (PH7.5); 2mM magnesium chloride; 0.5mM ethylene glycol diethyl ether diamine tetraacetic acid; 0.002% polyoxyethylene fatty alcohol ether; 1mM dithiothreitol and 1%DMSO;
2.检测溶液：TR-FRET Dilution Buffer；2. Detection solution: TR-FRET Dilution Buffer;
3.LRRK2人源重组蛋白：使用GST标签用杆状病毒在昆虫Sf9细胞中表达重组全长人LRRK2蛋白；3. LRRK2 human recombinant protein: using GST tag to express recombinant full-length human LRRK2 protein in insect Sf9 cells with baculovirus;
4.底物：0.4μM Fluorescein-ERM(LRRKtide)peptide；57μM ATP。4. Substrate: 0.4 μM Fluorescein-ERM (LRRKtide) peptide; 57 μM ATP.
检测方法:Detection method:
均相时间分辨荧光技术(HTRF)；Homogeneous Time-Resolved Fluorescence (HTRF);
Fluorescein-ERM(LRRKtide)peptide多肽与
Figure PCTCN2022129958-appb-000111
Tb-pERM(pLRRKtide)Antibody抗体在485nM和520nM之间的能量共振转移。 Fluorescein-ERM (LRRKtide) peptide peptide and
Figure PCTCN2022129958-appb-000111
Energy resonance transfer of Tb-pERM(pLRRKtide) Antibody between 485nM and 520nM.
实验操作:Experimental operation:
1.通过Echo550非接触式纳升级声波移液***加入待测化合物的DMSO溶液；1. Add the DMSO solution of the compound to be tested through the Echo550 non-contact nano-liter acoustic pipetting system;
2.用新鲜制备的反应溶液配置酶和多肽混合溶液，加入到反应孔穴中，室温下预温育20分钟；2. Prepare the enzyme and peptide mixed solution with the freshly prepared reaction solution, add it to the reaction well, and pre-incubate at room temperature for 20 minutes;
3.加入57μM ATP引发反应，室温反应90分钟；3. Add 57μM ATP to initiate the reaction, and react at room temperature for 90 minutes;
4.加入检测***(Fluorescein-ERM(LRRKtide)peptide多肽，
Figure PCTCN2022129958-appb-000112
Tb-pERM(pLRRKtide)Antibody抗体及10mM乙二胺四乙酸)，室温反应60分钟，用Em/Ex 520/485检测荧光信号； 4. Add the detection system (Fluorescein-ERM (LRRKtide) peptide polypeptide,
Figure PCTCN2022129958-appb-000112
Tb-pERM (pLRRKtide) Antibody antibody and 10mM ethylenediaminetetraacetic acid), reacted at room temperature for 60 minutes, and detected fluorescence signal with Em/Ex 520/485;
5.通过信号比值计算相对DMSO空白的相对酶活性抑制，利用软件XLfit5拟合曲线计算IC 50值。 5. The relative enzyme activity inhibition relative to DMSO blank was calculated by the signal ratio, and the IC 50 value was calculated by using the software XLfit5 to fit the curve.
实验结果：Experimental results:
表1 LRRK2激酶抑制活性测试结果Table 1 LRRK2 kinase inhibitory activity test results
供试品testing sample LRRK2激酶抑制活性(nM)LRRK2 kinase inhibitory activity (nM) 供试品testing sample LRRK2激酶抑制活性(nM)LRRK2 kinase inhibitory activity (nM)
化合物1Compound 1 66 化合物21Compound 21 66
化合物2Compound 2 33 化合物22Compound 22 55
化合物3Compound 3 66 化合物23Compound 23 66
化合物4Compound 4 77 化合物24Compound 24 55
化合物5Compound 5 55 化合物25Compound 25 55
化合物6Compound 6 55 化合物26Compound 26 55
化合物7 Compound 7 33 化合物27Compound 27 44
化合物8Compound 8 44 化合物28Compound 28 1515
化合物9Compound 9 44 化合物29Compound 29 1414
化合物10Compound 10 33 化合物30Compound 30 55
化合物11Compound 11 1414 化合物31Compound 31 1010
化合物12Compound 12 1212 化合物32Compound 32 77
化合物15Compound 15 1616 化合物33Compound 33 1313
化合物17Compound 17 55 化合物36Compound 36 1010
化合物18Compound 18 44 化合物37Compound 37 77
化合物19Compound 19 66 化合物39Compound 39 1616
化合物20Compound 20 44 // //
实验例2：体外评价LRRK2细胞(pSer935)抑制活性Experimental Example 2: In vitro evaluation of LRRK2 cell (pSer935) inhibitory activity
细胞准备：Cell preparation:
1.细胞解冻1. Cell Thawing
将293T细胞从液氮中取出，放入37摄氏度的水中。待冰完全融化后，将细胞转移至5毫升温暖培养液内，离心，弃去上清液，并把悬浮的细胞作新的细胞在培养基中培养。Take the 293T cells out of the liquid nitrogen and place them in water at 37°C. After the ice has completely melted, transfer the cells to 5 ml of warm culture medium, centrifuge, discard the supernatant, and culture the suspended cells as new cells in the culture medium.
2.细胞的培养和传代2. Cell culture and passage
将293T细胞在培养基里培养两至三天Culture 293T cells in medium for two to three days
3.细胞冷冻3. Cell Freezing
将培养好的细胞株放入新鲜的培养液中并稀释浓度至1*10^7，然后与等量的培养液混合。等分成每份1mL，置于零下80度一天，转移至液氮中保存。Put the cultured cell lines into fresh culture medium and dilute the concentration to 1*10^7, then mix with the same amount of culture medium. Divide into 1mL portions, store at minus 80 degrees for one day, and transfer to liquid nitrogen for storage.
实验步骤:Experimental steps:
1.(第一天)293T细胞播种1. (Day 1) Seeding of 293T cells
种植1.4×10^6/293T细胞于一块培养板上，两天的培养后，细胞数量可以成长为5×10^6，所以种子N+1块板子足够N个96孔板的实验。Plant 1.4×10^6/293T cells on a culture plate, and after two days of culture, the number of cells can grow to 5×10^6, so seeding N+1 plates is enough for N 96-well plate experiments.
2.(第二天)转染293T细胞2. (Second day) Transfection of 293T cells
一、添加5微升0.5微克/微升pcmv-flag-lrrk2到145微升DMEM培养液中，用吸管调匀；1. Add 5 microliters of 0.5 micrograms/microliter pcmv-flag-lrrk2 to 145 microliters of DMEM culture medium, and mix thoroughly with a pipette;
二、添加15微升转染试剂，吸管调匀；2. Add 15 microliters of transfection reagent, pipette and mix thoroughly;
三、室温平衡10分钟；3. Equilibrate at room temperature for 10 minutes;
四、添加0.5毫升预热的细胞培养基，调匀；4. Add 0.5 ml of preheated cell culture medium and mix thoroughly;
五、滴加650微升混合物到6孔板中，搅拌彻底；5. Add 650 microliters of the mixture dropwise to the 6-well plate and stir thoroughly;
六、培养板在37摄氏度含5％二氧化碳的加湿孵化器中培养20–24小时。6. Incubate the plate in a humidified incubator containing 5% carbon dioxide at 37°C for 20–24 hours.
3.(第三天)293T细胞种植到96孔板上3. (Day 3) 293T cells were planted on a 96-well plate
4.(第4天)抑制剂处理4. (Day 4) Inhibitor Treatment
一、将化合物用离心机处理；1. Process the compound with a centrifuge;
二、添加55微升细胞培养基到抑制剂板中。把盘子放在37摄氏度预热；2. Add 55 microliters of cell culture medium to the inhibitor plate. Preheat the plate at 37°C;
三、转移50微升含抑制剂的细胞培养至细胞培养板；3. Transfer 50 microliters of inhibitor-containing cell culture to the cell culture plate;
四、培养板在37摄氏度含5％二氧化碳的加湿孵化器中培养20–24小时；4. Cultivate the culture plate in a humidified incubator containing 5% carbon dioxide at 37 degrees Celsius for 20-24 hours;
五、用滴管抽取300微升含有抑制剂的培养液，取其中200微升，向其中加入100微升的分解剂，密封板子后于4摄氏度摇30分钟；5. Use a dropper to draw 300 microliters of the culture solution containing the inhibitor, take 200 microliters of it, add 100 microliters of the decomposition agent to it, seal the plate and shake it at 4 degrees Celsius for 30 minutes;
六、板子于零下20摄氏度保存直至使用。6. Store the board at minus 20 degrees Celsius until use.
5.(第5天)MSD程序5. (Day 5) MSD program
一、添加2微克/25微升/标记抗体到MSD板中，孵育2小时，(50微升3.9微克/微升Flag抗体+2.5毫升胎牛血清/每盘)。离心10秒(1000rpm)；1. Add 2 μg/25 μl/labeled antibody to the MSD plate and incubate for 2 hours (50 μl 3.9 μg/μl Flag antibody + 2.5 ml fetal bovine serum/per plate). Centrifuge for 10 seconds (1000rpm);
二、丢弃标志抗体，用300微升/洗涤缓冲液低速多点洗涤两次；2. Discard the marker antibody and wash twice with 300 μl/wash buffer at low speed;
三、添加50微升/块缓冲液、孵化2小；3. Add 50 μl/block buffer and incubate for 2 hours;
四、丢弃缓冲液，以300微升/洗涤缓冲液手动洗2次；4. Discard the buffer and wash manually twice with 300 μl/wash buffer;
五、转移12.5微升裂解缓冲液和细胞裂解液12.5微升至MSD板，室温孵育1小时；5. Transfer 12.5 microliters of lysis buffer and 12.5 microliters of cell lysate to the MSD plate, and incubate at room temperature for 1 hour;
六、丢弃裂解液，用300微升/洗涤缓冲液多点、低速度洗3次；6. Discard the lysate and wash 3 times with 300 microliters/wash buffer at low speed;
七、稀释ps935(1:200)抗，加入25微升/抗体，室温培育1小时；7. Dilute the ps935 (1:200) antibody, add 25 microliters/antibody, and incubate at room temperature for 1 hour;
八、弃第一抗体，用300微升/洗涤缓冲液多点、低速度洗3次；8. Discard the primary antibody and wash 3 times with 300 microliters/wash buffer at low speed;
九、稀释山羊抗兔抗体1:500，添加25微升/抗体至孵育板，室温培养1小时；9. Dilute goat anti-rabbit antibody 1:500, add 25 microliters/antibody to the incubation plate, and incubate at room temperature for 1 hour;
十、弃第二抗体，用300微升/洗涤缓冲液多点、低速度洗3次。最后一次的洗涤缓冲液到MSD阅读器；10. Discard the secondary antibody and wash 3 times with 300 microliters/wash buffer at low speed. The last wash buffer to the MSD reader;
十一、采集两次数据；11. Collect data twice;
十二、放弃最后一次的清洗液，添加150微升/2倍缓冲液至待读取孔板；12. Discard the last washing solution and add 150 microliters/2 times buffer solution to the well plate to be read;
十三、孵化后约3分钟，于15分钟读取数据。13. About 3 minutes after incubation, read the data at 15 minutes.
实验结果：Experimental results:
表2 pSer935细胞抑制活性测试结果Table 2 pSer935 cell inhibitory activity test results
Figure PCTCN2022129958-appb-000113
Figure PCTCN2022129958-appb-000113
Figure PCTCN2022129958-appb-000114
Figure PCTCN2022129958-appb-000114
结论：本发明化合物具有显著的LRRK2酶和细胞(pSer935)抑制活性。Conclusion: The compounds of the present invention have significant LRRK2 enzyme and cell (pSer935) inhibitory activity.
实验例3：化合物药代动力学评价Experimental Example 3: Pharmacokinetic Evaluation of Compounds
实验目的：研究化合物在C57BL/6小鼠体内药代动力学及脑组织和血浆药物浓度比Experimental purpose: To study the pharmacokinetics of the compound in C57BL/6 mice and the ratio of drug concentration in brain tissue to plasma
实验材料：C57BL/6小鼠(雄性，8周龄，体重25g-30g)Experimental material: C57BL/6 mice (male, 8 weeks old, body weight 25g-30g)
实验操作：以标准方案测试化合物静脉注射(IV)及口服(PO)给药后的啮齿类动物药代特征，实验中候选化合物配成1mg/mL澄清溶液，给予小鼠单次静脉注射及口服给药。静注及口服溶媒均为5％DMSO/5％15-羟基硬脂酸聚乙二醇酯(Solutol)/90％水溶液。首先使用4只雄性C57BL/6小鼠，两只小鼠进行静脉注射给药，给药剂量为1mg/kg，收集给药后0.0833，0.25，0.5，1，2，4，6，8，24h的血浆样品，另外两只小鼠口服灌胃给药，给药剂量为10mg/kg，收集给药后0.25，0.5，1，2，4，6，8，24h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，加入含内标的乙腈溶液沉淀蛋白，充分混匀离心取上清液进样，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)等。另外，再使用6只小鼠口服灌胃给药，给药剂量为5mg/kg或者10mg/kg，在给药后0.5，2和8小时，将收集全脑。将组织样品用15mM胎牛血清[胎牛血清(pH＝7.4)缓冲液：甲醇(体积比，2：1)]匀浆，均质比为1：5(w：v)，并将匀浆液分成2个等分样品，一个用于分析，另一个用于备份。另外，收集给药后0.25，0.5，1，2，4，8h的血浆，血浆样品在收集半小时内，通过在约4℃，3000g，15分钟离心处理分离上清得血浆样品。将血浆样品储存在聚丙烯管中，在干冰上快速冷冻并保持在-80℃直至LC/MS/MS分析。加入含内标的乙腈溶液沉淀蛋白，充分混匀离心取上清液进样，以LC-MS/MS分析方法定量分析血药浓度，并计算在0.5小时脑组织和血浆药物浓度比例(B/P)。 Experimental operation: The pharmacokinetic characteristics of rodents after intravenous (IV) and oral (PO) administration of the compound were tested according to the standard protocol. In the experiment, the candidate compound was formulated into a 1 mg/mL clear solution, and given to mice for a single intravenous injection and oral administration medication. Both intravenous and oral vehicles are 5% DMSO/5% polyethylene glycol 15-hydroxystearate (Solutol)/90% aqueous solution. Firstly, four male C57BL/6 mice were used, and two mice were administered intravenously at a dose of 1 mg/kg, and collected at 0.0833, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration The plasma samples of the other two mice were administered orally by gavage at a dose of 10 mg/kg, and plasma samples were collected at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration. Collect whole blood samples within 24 hours, centrifuge at 3000g for 15 minutes, separate supernatant to obtain plasma samples, add acetonitrile solution containing internal standard to precipitate protein, mix well and centrifuge to take supernatant for injection, and quantify by LC-MS/MS analysis method Analyze blood drug concentration and calculate pharmacokinetic parameters, such as peak concentration (C max ), clearance (CL), half-life (T 1/2 ), tissue distribution (Vdss), area under the drug-time curve (AUC 0- last ), bioavailability (F), etc. In addition, 6 more mice were used for oral gavage administration at a dosage of 5 mg/kg or 10 mg/kg, and the whole brains were collected at 0.5, 2 and 8 hours after administration. The tissue sample was homogenized with 15mM fetal bovine serum [fetal bovine serum (pH=7.4) buffer: methanol (volume ratio, 2:1)], the homogenization ratio was 1:5 (w:v), and the homogenate Divide into 2 aliquots, one for analysis and the other for backup. In addition, the plasma samples were collected at 0.25, 0.5, 1, 2, 4, and 8 hours after administration. Within half an hour of collecting the plasma samples, the supernatant was separated by centrifugation at about 4° C., 3000 g, for 15 minutes to obtain plasma samples. Plasma samples were stored in polypropylene tubes, snap frozen on dry ice and kept at -80°C until LC/MS/MS analysis. Add the acetonitrile solution containing the internal standard to precipitate the protein, mix well and centrifuge to take the supernatant for sample injection, quantitatively analyze the blood drug concentration by LC-MS/MS analysis method, and calculate the concentration ratio of the brain tissue and plasma drug in 0.5 hours (B/P ).
本发明实施例在小鼠体内的药代动力学相关参数如下表3所示。The relevant parameters of the pharmacokinetics of the embodiments of the present invention in mice are shown in Table 3 below.
表3体内药代动力学测试结果Table 3 in vivo pharmacokinetic test results
Figure PCTCN2022129958-appb-000115
Figure PCTCN2022129958-appb-000115
Figure PCTCN2022129958-appb-000116
Figure PCTCN2022129958-appb-000116
注：“(B/P)@0.5h”指代是单点(0.5小时)的脑血比。Note: "(B/P)@0.5h" refers to the brain blood ratio of a single point (0.5 hour).
结论：本发明化合物具有良好的体内药代动力学性质，包括良好的脑组织和血浆药物浓度比例(B/P)等。Conclusion: the compound of the present invention has good in vivo pharmacokinetic properties, including good brain tissue and plasma drug concentration ratio (B/P).
实验例4：体内药效实验Experimental Example 4: In Vivo Drug Efficacy Experiment
实验目的：测试化合物对小鼠脑组织LRRK2磷酸化抑制效果。Experimental purpose: To test the inhibitory effect of compounds on LRRK2 phosphorylation in mouse brain tissue.
实验材料：Experimental Materials:
动物：C57BL/6J小鼠(雄性，6～7周龄，体重20g-22g)。Animals: C57BL/6J mice (male, 6-7 weeks old, body weight 20g-22g).
实验过程：experiment procedure:
1.实验分组：所有小鼠在实验前一天称取体重，按照体重随机分为5组，每组4只，体重过大和过小的小鼠剔除，保证每组的平均体重一致。实验分组如下表4所示，共5组，即空白组，阳性对照药物(组-2～3,n＝4,MLi-2和DNL151),受试药物实施例(组3～5,n＝4)。1. Experimental grouping: All mice were weighed the day before the experiment, and were randomly divided into 5 groups according to their body weight, with 4 mice in each group. The overweight and underweight mice were eliminated to ensure that the average weight of each group was the same. The experimental groupings are shown in Table 4 below, with 5 groups in total, i.e. blank group, positive control drug (group-2～3, n=4, MLi-2 and DNL151), test drug embodiment (group 3-5, n= 4).
表4实验分组和剂量设计Table 4 Experimental grouping and dose design
Figure PCTCN2022129958-appb-000117
Figure PCTCN2022129958-appb-000117
溶媒：5％DMSO/聚乙二醇十二羟基硬脂酸酯(
Figure PCTCN2022129958-appb-000118
HS 15)/90％超纯水 Vehicle: 5% DMSO/polyethylene glycol lauryl hydroxystearate (
Figure PCTCN2022129958-appb-000118
HS 15)/90% ultrapure water
2.实验操作：2. Experimental operation:
2.1给药和组织收集2.1 Administration and tissue collection
各组动物按照表4中设计按时间顺序依次口服给予溶媒或待测化合物，给药1小时后处死，冰上取大脑皮层，置于1.5mLEP管中，加入400μLPIPA蛋白裂解液，同时加入4μLPMSF、4μL磷酸酶抑制剂A和4μL磷酸酶抑制剂B，用剪刀搅碎，超声破碎，冰上静置15min，4℃离心15min，取上清，冰上放置。取2μL上清，BCA蛋白浓度测定。根据上清体积加入相应比例的5×蛋白上样缓冲液；蛋白变性，干浴锅100℃，10min。低速离心，-80℃保存。The animals in each group were orally administered with the vehicle or the compound to be tested in chronological order according to the design in Table 4, and were executed 1 hour after the administration. 4 μL of phosphatase inhibitor A and 4 μL of phosphatase inhibitor B were crushed with scissors, ultrasonically broken, left on ice for 15 min, centrifuged at 4°C for 15 min, supernatant was taken, and placed on ice. Take 2 μL of the supernatant, and measure the BCA protein concentration. According to the supernatant volume, add the corresponding proportion of 5× protein loading buffer; protein denaturation, dry bath 100°C, 10min. Centrifuge at low speed and store at -80°C.
2.2药效检测2.2 Drug efficacy test
(1)配置新鲜电泳缓冲液，上样量50μg，跑胶，使用预染的Marker，电压60-100V，时间1.5h左右(根据目的蛋白分子量大小决定)。(1) Prepare fresh electrophoresis buffer, load 50 μg of sample, run gel, use pre-stained marker, voltage 60-100V, time about 1.5h (determined according to the molecular weight of the target protein).
(2)配转移缓冲液(使用10倍的转移缓冲液稀释，加甲醇15-20％,甲醇浓度与目的蛋白大小有关)。(2) Prepare transfer buffer (dilute with 10 times transfer buffer, add 15-20% methanol, the concentration of methanol is related to the size of the target protein).
(3)PVDF膜用100％甲醇处理30s左右(可延长)，变为灰色，之后用转移缓冲液浸泡，备用；同时需浸泡的还有滤纸和垫片(之后保持膜湿润)。(3) PVDF membrane is treated with 100% methanol for about 30s (can be extended), and turns gray, and then soaked in transfer buffer for later use; at the same time, filter paper and gaskets need to be soaked (keep the membrane moist afterwards).
(4)叠放顺序：黑色板最下面，垫片，两层滤纸，胶(反向放)，PVDF膜(与胶接触面左下角做好标记)，两层滤纸，垫片，白色板最上面，扣板(每层用玻璃棒赶气泡)。(4) Stacking sequence: the bottom of the black board, gasket, two layers of filter paper, glue (reversely placed), PVDF membrane (mark the lower left corner of the contact surface with the glue), two layers of filter paper, gasket, and white board at the end On the top, gusset (every layer uses a glass rod to catch air bubbles).
(5)准备冰，装好转膜装置，电泳100V，时间1.5-2h(注意冰量要足够，可以冰里加点水方便传热，转膜会释放大量热量)。(5) Prepare ice, install the membrane transfer device, run electrophoresis at 100V for 1.5-2 hours (note that the amount of ice should be sufficient, you can add some water to the ice to facilitate heat transfer, and membrane transfer will release a lot of heat).
(6)用1×TBST配制5％脱脂牛奶(1×TBST＝1×TBS加0.1％Tween)。(6) Prepare 5% skimmed milk with 1×TBST (1×TBST=1×TBS plus 0.1% Tween).
(7)转膜完毕，若Marker转上，则说明转膜成功。(7) After the film transfer is completed, if the Marker is turned on, it means that the film transfer is successful.
(8)用5％脱脂牛奶，室温封闭2h。(8) Block with 5% skimmed milk for 2 hours at room temperature.
(9)用一抗孵育，4℃过夜。(9) Incubate with primary antibody overnight at 4°C.
(10)用TBST洗膜10min×4次。(10) Wash the membrane with TBST for 10 min×4 times.
(11)用二抗孵育1h(用荧光二抗则需避光)。(11) Incubate with secondary antibody for 1 h (when fluorescent secondary antibody is used, it needs to be protected from light).
(12)用1×TBST洗膜20min×3次。(12) Wash the membrane with 1×TBST for 20 min×3 times.
(13)显色，ECL化学发光。(13) Color development, ECL chemiluminescence.
3.实验结果3. Experimental results
3.1脑组织免疫印迹实验(Western Blot)结果见图1。3.1 The results of Western Blot of brain tissue are shown in Figure 1.
结论：本发明化合物对在小鼠脑组织中LRRK2磷酸化水平有明显地抑制作用。Conclusion: the compound of the present invention has obvious inhibitory effect on the phosphorylation level of LRRK2 in mouse brain tissue.

Claims (15)

  1. 式(III-1)、(II-2)、(II-3)、(II-4)和(III-2)所示化合物或其药学上可接受的盐：Compounds represented by formulas (III-1), (II-2), (II-3), (II-4) and (III-2) or pharmaceutically acceptable salts thereof:
    Figure PCTCN2022129958-appb-100001
    Figure PCTCN2022129958-appb-100001
    其中，in,
    A选自
    Figure PCTCN2022129958-appb-100002
    Figure PCTCN2022129958-appb-100003
    A selected from
    Figure PCTCN2022129958-appb-100002
    Figure PCTCN2022129958-appb-100003
    结构单元
    Figure PCTCN2022129958-appb-100004
    选自
    Figure PCTCN2022129958-appb-100005
    Structural units
    Figure PCTCN2022129958-appb-100004
    selected from
    Figure PCTCN2022129958-appb-100005
    X 1为CH，X 2为C-A；或者，X 1为C-A，X 2为CH； X 1 is CH, X 2 is CA; or, X 1 is CA, X 2 is CH;
    Y为-C(R 9R 10)-或-O-； Y is -C(R 9 R 10 )- or -O-;
    Z 1为CH，Z 2为N；或者，Z 1为N，Z 2为CH； Z 1 is CH, Z 2 is N; or, Z 1 is N, Z 2 is CH;
    R 1和R 2分别独立地选自H、F、Cl、Br、I、CH 3和CF 3； R 1 and R 2 are independently selected from H, F, Cl, Br, I, CH 3 and CF 3 ;
    R 3和R 4分别独立地选自H和C 1-3烷基，所述C 1-3烷基任选被1、2或3个卤素取代； R 3 and R 4 are independently selected from H and C 1-3 alkyl, and the C 1-3 alkyl is optionally substituted by 1, 2 or 3 halogens;
    R 5和R 6分别独立地选自H、F、CH 3和CF 3； R 5 and R 6 are independently selected from H, F, CH 3 and CF 3 ;
    R 7选自CH 3和CHF 2； R 7 is selected from CH 3 and CHF 2 ;
    R 9和R 10分别独立地选自H、F、Cl、Br、I、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基分别独立地任选被1、2或3个卤素取代； R 9 and R 10 are independently selected from H, F, Cl, Br, I, C 1-3 alkyl and C 1-3 alkoxy, the C 1-3 alkyl and C 1-3 alkoxy Each group is independently optionally substituted by 1, 2 or 3 halogens;
    m、n、q、t分别独立地选自0、1和2。m, n, q, t are independently selected from 0, 1 and 2, respectively.
  2. 根据权利要求1所述的化合物、其立体异构体或其药学上可接受的盐，其选自式(II-1)、(II-2)、(II-3)和(II-4)所示化合物：The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, selected from the group consisting of formulas (II-1), (II-2), (II-3) and (II-4) Compound shown:
    Figure PCTCN2022129958-appb-100006
    Figure PCTCN2022129958-appb-100006
    其中，in,
    A选自
    Figure PCTCN2022129958-appb-100007
    A selected from
    Figure PCTCN2022129958-appb-100007
    X 1为CH，X 2为C-A；或者，X 1为C-A，X 2为CH； X 1 is CH, X 2 is CA; or, X 1 is CA, X 2 is CH;
    Y为-C(R 9R 10)-或-O-； Y is -C(R 9 R 10 )- or -O-;
    Z 1为CH，Z 2为N；或者，Z 1为N，Z 2为CH； Z 1 is CH, Z 2 is N; or, Z 1 is N, Z 2 is CH;
    R 1和R 2分别独立地选自H、F、Cl、Br、I、CH 3和CF 3； R 1 and R 2 are independently selected from H, F, Cl, Br, I, CH 3 and CF 3 ;
    R 3和R 4分别独立地选自H和C 1-3烷基，所述C 1-3烷基任选被1、2或3个卤素取代； R 3 and R 4 are independently selected from H and C 1-3 alkyl, and the C 1-3 alkyl is optionally substituted by 1, 2 or 3 halogens;
    R 5和R 6分别独立地选自H和F； R 5 and R 6 are independently selected from H and F;
    R 7选自CH 3和CHF 2； R 7 is selected from CH 3 and CHF 2 ;
    R 9和R 10分别独立地选自H、F、Cl、Br、I、C 1-3烷基和C 1-3烷氧基，所述C 1-3烷基和C 1-3烷氧基分别独立地任选被1、2或3个卤素取代； R 9 and R 10 are independently selected from H, F, Cl, Br, I, C 1-3 alkyl and C 1-3 alkoxy, the C 1-3 alkyl and C 1-3 alkoxy Each group is independently optionally substituted by 1, 2 or 3 halogens;
    m、n、q、t分别独立地选自0、1和2。m, n, q, t are independently selected from 0, 1 and 2, respectively.
  3. 根据权利要求1所述的化合物、其立体异构体或其药学上可接受的盐，其选自The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, which is selected from
    Figure PCTCN2022129958-appb-100008
    Figure PCTCN2022129958-appb-100008
    其中，A、R 1、R 2、R 3、R 6和R 7如权利要求1所定义。 Wherein, A, R 1 , R 2 , R 3 , R 6 and R 7 are as defined in claim 1.
  4. 根据权利要求2所述的化合物或其药学上可接受的盐，其中，化合物如式(I-1)、(I-2)、(I-3)、(I-4)、(I-5)和(I-6)所示：The compound or pharmaceutically acceptable salt thereof according to claim 2, wherein the compound is such as formula (I-1), (I-2), (I-3), (I-4), (I-5 ) and (I-6):
    Figure PCTCN2022129958-appb-100009
    Figure PCTCN2022129958-appb-100009
    其中，Y、R 1、R 2、R 3、R 4、m、n、q和t如权利要求2所定义。 Wherein, Y, R 1 , R 2 , R 3 , R 4 , m, n, q and t are as defined in claim 2.
  5. 根据权利要求1-4任意一项所述的化合物或其药学上可接受的盐，其中，R 1选自H和F。 The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-4, wherein R is selected from H and F.
  6. 根据权利要求1-4任意一项所述的化合物或其药学上可接受的盐，其中，R 2选自Cl和F。 The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-4, wherein R is selected from Cl and F.
  7. 根据权利要求1-4任意一项所述的化合物或其药学上可接受的盐，其中，R 3和R 4分别独立地选自 CH 3。 The compound according to any one of claims 1-4 or a pharmaceutically acceptable salt thereof, wherein R 3 and R 4 are independently selected from CH 3 .
  8. 根据权利要求1-4任意一项所述的化合物或其药学上可接受的盐，其中，R 9和R 10分别独立地选自H、F、CH 3、-OCHF 2和-OCH 3。 The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-4, wherein R 9 and R 10 are independently selected from H, F, CH 3 , -OCHF 2 and -OCH 3 .
  9. 根据权利要求1-4或6任意一项所述的化合物或其药学上可接受的盐，其中，Y为-CHF-、-CF 2-、-CH 2-、-CH(OCH 3)-、-CH(OCHF 2)-和-O-。 The compound or pharmaceutically acceptable salt thereof according to any one of claims 1-4 or 6, wherein Y is -CHF-, -CF 2 -, -CH 2 -, -CH(OCH 3 )-, -CH(OCHF 2 )- and -O-.
  10. 根据权利要求1-4任意一项所述的化合物或其药学上可接受的盐，其中，结构单元
    Figure PCTCN2022129958-appb-100010
    选自
    Figure PCTCN2022129958-appb-100011
    Figure PCTCN2022129958-appb-100012
    The compound or pharmaceutically acceptable salt thereof according to any one of claims 1-4, wherein the structural unit
    Figure PCTCN2022129958-appb-100010
    selected from
    Figure PCTCN2022129958-appb-100011
    Figure PCTCN2022129958-appb-100012
  11. 根据权利要求1-3任意一项所述的化合物或其药学上可接受的盐，其中，结构单元
    Figure PCTCN2022129958-appb-100013
    选自
    Figure PCTCN2022129958-appb-100014
    The compound or pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein the structural unit
    Figure PCTCN2022129958-appb-100013
    selected from
    Figure PCTCN2022129958-appb-100014
  12. 根据权利要求1-3任意一项所述的化合物或其药学上可接受的盐，其中，结构单元
    Figure PCTCN2022129958-appb-100015
    选自
    Figure PCTCN2022129958-appb-100016
    The compound or pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein the structural unit
    Figure PCTCN2022129958-appb-100015
    selected from
    Figure PCTCN2022129958-appb-100016
  13. 根据权利要求1所述的化合物或其药学上可接受的盐，其中，上述式(II-2)中的结构单元
    Figure PCTCN2022129958-appb-100017
    选自
    Figure PCTCN2022129958-appb-100018
    The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit in the above formula (II-2)
    Figure PCTCN2022129958-appb-100017
    selected from
    Figure PCTCN2022129958-appb-100018
    或者，上述式(III-2)中的结构单元
    Figure PCTCN2022129958-appb-100019
    选自
    Figure PCTCN2022129958-appb-100020
    Alternatively, the structural unit in the above formula (III-2)
    Figure PCTCN2022129958-appb-100019
    selected from
    Figure PCTCN2022129958-appb-100020
  14. 化合物或其药学上可接受的盐，其中，化合物选自A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from
    Figure PCTCN2022129958-appb-100021
    Figure PCTCN2022129958-appb-100021
    Figure PCTCN2022129958-appb-100022
    Figure PCTCN2022129958-appb-100022
    Figure PCTCN2022129958-appb-100023
    Figure PCTCN2022129958-appb-100023
    Figure PCTCN2022129958-appb-100024
    Figure PCTCN2022129958-appb-100024
    Figure PCTCN2022129958-appb-100025
    Figure PCTCN2022129958-appb-100025
  15. 根据权利要求1～14任意一项所述的化合物或其药学上可接受的盐在制备治疗帕金森病药物中的应用。The use of the compound according to any one of claims 1-14 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating Parkinson's disease.
PCT/CN2022/129958 2021-11-05 2022-11-04 Azaspiro compound WO2023078411A1 (en)

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WO2016000581A1 (en) * 2014-07-04 2016-01-07 南京明德新药研发股份有限公司 Spirocyclic aryl phosphorus oxide and aryl phosphorus sulfide
WO2017086832A1 (en) * 2015-11-19 2017-05-26 Акционерное Общество "Р-Фарм" (Ао "Р-Фарм") Substituted n2-(4-amino-2-methoxyphenyl)-n4-[2-(dimethylphosphinoyl)-phenyl]-5-chloro-pyrimidine-2,4-diamines as alk and egfr modulators intended for treating cancer
WO2019201334A1 (en) * 2018-04-20 2019-10-24 南京明德新药研发有限公司 Dimethylphosphine oxide compound
WO2021073600A1 (en) * 2019-10-18 2021-04-22 贵州伊诺其尼科技有限公司 Crystal form of lrrk2 inhibitor and preparation method therefor
WO2021125803A1 (en) * 2019-12-16 2021-06-24 한국화학연구원 Novel pyrimidin derivative and use thereof

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009127642A2 (en) * 2008-04-15 2009-10-22 Cellzome Limited Use of lrrk2 inhibitors for neurodegenerative diseases
WO2016000581A1 (en) * 2014-07-04 2016-01-07 南京明德新药研发股份有限公司 Spirocyclic aryl phosphorus oxide and aryl phosphorus sulfide
WO2017086832A1 (en) * 2015-11-19 2017-05-26 Акционерное Общество "Р-Фарм" (Ао "Р-Фарм") Substituted n2-(4-amino-2-methoxyphenyl)-n4-[2-(dimethylphosphinoyl)-phenyl]-5-chloro-pyrimidine-2,4-diamines as alk and egfr modulators intended for treating cancer
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WO2021125803A1 (en) * 2019-12-16 2021-06-24 한국화학연구원 Novel pyrimidin derivative and use thereof

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