WO2022089629A1 - 1,2,4-triazole derivative and preparation method therefor and use thereof - Google Patents

1,2,4-triazole derivative and preparation method therefor and use thereof Download PDF

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WO2022089629A1
WO2022089629A1 PCT/CN2021/127847 CN2021127847W WO2022089629A1 WO 2022089629 A1 WO2022089629 A1 WO 2022089629A1 CN 2021127847 W CN2021127847 W CN 2021127847W WO 2022089629 A1 WO2022089629 A1 WO 2022089629A1
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alkyl
compound
mmol
hydroxyl
substituted
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PCT/CN2021/127847
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Chinese (zh)
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秦引林
苏梅
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江苏柯菲平医药股份有限公司
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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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4196—1,2,4-Triazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present invention belongs to the technical field of medicine, and particularly relates to novel compounds, pharmaceutical combinations comprising the compounds, methods for preparing the compounds, and the use of the compounds in the treatment of various diseases, disorders or conditions related to abnormal cellular responses triggered by inappropriate nuclear transport. use.
  • the nucleus of eukaryotes has a double-layered membrane structure, and macromolecules need to be transported across the nuclear membrane through the mediation of nucleocytoplasmic transport receptors.
  • Specific proteins and RNAs are transported into and out of the nucleus by specific transport molecules, wherein those that transport the molecules into the nucleus are classified as nuclear import proteins, and those that transport molecules out of the nucleus are classified as nuclear export proteins.
  • nuclear export protein 1 exportin 1, XPO1
  • CCM1 chromosomal region stabilizing protein 1
  • XPO1 can mediate the transport of more than 200 kinds of proteins, including tumor suppressor proteins (TSPs), P53, P73, etc., anti-apoptotic proteins (IAPs), nucleophsmim (NPM), survivin And growth regulatory proteins (GRPs) P21 and P27.
  • TSPs tumor suppressor proteins
  • IAPs anti-apoptotic proteins
  • NPM nucleophsmim
  • GFPs survivin And growth regulatory proteins
  • XPO1 inhibitors can induce apoptosis in cancer cells, even in the presence of oncogenic activating signals or growth stimulating signals, without affecting normal (untransformed) cells.
  • the expression level of XPO1 in cancer cells is 2-4 times higher than that in normal cells on average, which makes most of TSPs, IAPs, and GRPs transported out of the nucleus, which has an adverse effect on tumor suppressor protein recognition and induction of cancer cell apoptosis, which makes cancer cells proliferate. Differentiation is not limited.
  • the first-generation XPO1 inhibitor selinexor showed high blood-brain barrier penetration and greater toxicity in preclinical and clinical studies, and a new generation of XPO1 inhibitors is urgently needed to meet clinical needs.
  • the present invention provides a 1,2,4-triazole derivative and a preparation method and application thereof.
  • One of the objects of the present invention is to provide a compound, characterized in that the compound is the compound represented by formula I, or its optical isomer, enantiomer, diastereomer, racemate or racemate a mixture, or a solvate, prodrug, or a pharmaceutically acceptable salt thereof;
  • R 2 and R 3 are each independently selected from H or C 1 -C 3 alkyl groups
  • R 4 is selected from H or C 1 -C 3 alkyl
  • R 2 , R 3 are each independently selected from H, methyl or ethyl.
  • R4 is selected from H, methyl or ethyl.
  • R1 is selected from unsubstituted or optionally substituted by A 5-6-membered heteroaryl;
  • R1 is selected from unsubstituted or optionally substituted 8-15-membered fused heteroaryl; substituents are selected from halogen, hydroxyl, -CN, C 1 -C 3 alkyl, C 1 -C 3 Alkoxy, hydroxy C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl.
  • R1 is selected from unsubstituted or optionally substituted pyrrolyl, furyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl , thiadiazolyl or oxadiazolyl. Substituents may be as described above.
  • R1 is an optionally substituted 6-membered heteroaryl having 1, 2 or 3 heteroatoms independently selected from the group consisting of nitrogen, oxygen and sulfur.
  • R1 is selected from unsubstituted or optionally substituted pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, or triazinyl. Substituents may be as described above.
  • the compound of the present invention is the compound represented by formula I-1, or its optical isomer, enantiomer, diastereomer, racemate or a racemic mixture, or a solvate, prodrug, or a pharmaceutically acceptable salt thereof;
  • R1 is as described above.
  • the compound is selected from the group consisting of:
  • the present invention also provides a compound, which is characterized in that it is a compound represented by formula II, or an optical isomer, enantiomer, diastereomer, racemate or racemic mixture thereof, or a solvate thereof drug, prodrug or a pharmaceutically acceptable salt thereof;
  • ring C is selected from 5-6 membered heteroaryl, 5-6 membered heterocyclyl, wherein said heteroaryl or heterocyclyl is optionally further substituted by one or more R 7 ; preferably, ring C is selected from from 5-6 membered heterocyclyl, wherein said heterocyclyl is optionally further substituted with one or two R7 ;
  • R 7 is selected from hydrogen atom, halogen, cyano, hydroxyl, C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, C 1 -C 8 alkoxy, C 1 -C 8 haloalkyl, C 3 ⁇ C 10 halogenated cycloalkyl, C 1 -C 8 halogenated alkoxy; preferably, R 7 is selected from hydrogen atom, halogen, cyano, hydroxyl, C 1 -C 3 alkyl, C 3 -C 6 cycloalkane base, C 1 -C 3 alkoxy, C 1 -C 3 haloalkyl, C 3 -C 6 halocycloalkyl, C 1 -C 3 haloalkoxy;
  • R 9 is selected from the following groups unsubstituted or optionally substituted by one or more E: hydroxyl, amino;
  • Substituent E is selected from: halogen, hydroxyl, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl , C 2 -C 10 heterocyclyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 3 -C 10 halocycloalkyl, Or form a 4-8 membered heterocycle with the substituted atom;
  • R 9 is selected from: hydrogen atom, hydroxyl, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 3 -C 10 halocycloalkyl, C 6 -C 10 aryl, C 2 -C 10 heteroaryl or C 2 -C 10 heterocyclic.
  • R 9 is selected from the following groups unsubstituted or optionally substituted by one or more E: hydroxyl, amino;
  • Substituent E is selected from: halogen, hydroxyl, C 1 -C 4 alkyl, C 3 -C 8 cycloalkyl , C 2 -C 8 heterocyclyl, C 1 -C 4 alkoxy, C 1 -C 4 haloalkyl, C 3 -C 8 halocycloalkyl, Or form a 4-6 membered heterocycle with the substituted atom;
  • R 9 is selected from: hydrogen atom, hydroxyl, C 1 -C 3 alkyl, C 3 -C 8 cycloalkyl, C 1 -C 3 alkane
  • the present invention also provides compounds represented by formula II-1 or formula II-2, or optical isomers, enantiomers, diastereomers, racemates or exoisomers thereof A racemic mixture, or a solvate, prodrug or a pharmaceutically acceptable salt thereof:
  • R 8 and R 9 are as previously described; in some more specific embodiments, R 8 is H or pyrimidine.
  • the present invention also provides compounds shown below, or optical isomers, enantiomers, diastereomers, racemates or racemic mixtures thereof, or solvates thereof , a prodrug or a pharmaceutically acceptable salt thereof:
  • the second object of the present invention is to provide a preparation method of the compound.
  • the third purpose of this paper is to provide a pharmaceutical composition
  • a pharmaceutical composition comprising the compounds represented by formula I and formula II, or optical isomers, enantiomers, diastereomers, racemates or racemic mixtures thereof, or a solvate, prodrug thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • the fourth purpose of this paper is to provide compounds represented by formula I and formula II, or optical isomers, enantiomers, diastereomers, racemates or racemic mixtures thereof, or solvates and prodrugs thereof , or a pharmaceutically acceptable salt thereof, or the application of the composition of the present invention as an XPO1 inhibitor.
  • the fifth purpose of this paper is to provide compounds represented by formula I and formula II, or optical isomers, enantiomers, diastereomers, racemates or racemic mixtures thereof, or solvates and prodrugs thereof , or a pharmaceutically acceptable salt thereof, or the composition of the present invention as a use in the manufacture of a medicament for the treatment of various diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport.
  • the compounds represented by formula I and formula II of the present invention are also useful for the treatment of various diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport.
  • one embodiment of the present invention is the use of a compound of the present invention, or a pharmaceutically acceptable salt thereof, for the treatment of various diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport.
  • the compounds provided by the present invention are also useful for the study of nuclear transport regulation in biological and pathological phenomena, such as the study of kinase-mediated intracellular signal transduction pathways and the comparative evaluation of novel nuclear transport modulators.
  • various diseases, disorders or conditions described herein that are associated with abnormal cellular responses triggered by inappropriate nuclear transport include, but are not limited to, proliferative disorders, cancers, inflammatory disorders, autoimmune disorders, viruses Infections, neurodegenerative disorders, ophthalmic disorders, disorders of abnormal tissue growth, disorders related to food intake, allergies, and respiratory disorders.
  • Also provided herein is a method for treating a patient having a disease, disorder or condition associated with various inappropriate nuclear transport-triggered abnormal cellular responses, comprising administering to the patient a therapeutically effective amount of formula I as defined herein
  • the patient is a human.
  • halogen means -F (sometimes referred to herein as “fluoro"), -Cl, -Br and -I.
  • C1-C3 alkyl refers to groups having one to three, one to six, two to six, respectively A saturated linear or branched monovalent hydrocarbon radical of one or three to six carbon atoms. Examples include, but are not limited to, methyl, ethyl, 1-propyl, isopropyl, 1-butyl, isobutyl, sec-butyl, tert-butyl, 2-methyl-2-propyl, pentyl base, neopentyl and hexyl.
  • C1-C6 alkoxy refers to a saturated straight or branched monovalent alkoxy group having one to six carbon atoms, wherein the bond is on an oxygen atom. Examples include methoxy, ethoxy, propoxy, isopropoxy, butoxy and tert-butoxy.
  • hydroxy C1-C6 alkyl- refers to a saturated straight or branched chain monovalent alkyl group having one to six carbon atoms, wherein at least one carbon atom is substituted with a hydroxyl group.
  • haloC1-C6alkyl- refers to a saturated straight or branched monovalent alkyl group having one to six carbon atoms, wherein at least one carbon atom is substituted with a halogen.
  • cyano C1-C6 alkyl- refers to a saturated straight or branched monovalent alkyl group having one to six carbon atoms, one of which is replaced by a cyano group.
  • C3-C6 cycloalkyl refers to cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
  • heterocycle refers to a monocyclic or bicyclic non-aromatic heterocycle containing, in addition to carbon atoms, 1-4 heteroatoms selected from the group consisting of oxygen atoms, sulfur atoms and nitrogen atoms, as Specific examples include azetidine, pyrrolidine, pyrazolidine, piperidine, oxetane, tetrahydrofuran, tetrahydropyran, tetrahydrothiophene, dihydroimidazole, imidazolidine ( imidazolidine), tetrahydropyrazine, piperazine, morpholine, etc.
  • the 5-15 membered heterocycloalkyl is a 5-15 membered heterocycloalkyl having 1-2 heteroatoms independently optionally selected from N, S, or O.
  • the 5-10 membered heterocycloalkyl is a 5-10 membered heterocycloalkyl having 1-2 heteroatoms independently optionally selected from N, S, or O.
  • heteroaryl represents a stable monocyclic ring of up to 3-15 atoms in the ring or a bicyclic carbocyclic ring of up to 3-15 atoms in each ring, wherein at least one ring is aromatic and contains 1-4 A heteroatom selected from O, N and S.
  • Heteroaryl groups within the scope of this definition include, but are not limited to, acridinyl, carbazolyl, cinnolinyl, quinoxolinyl, pyrazolyl, indolyl, benzotriazolyl, furyl, thienyl, benzene thienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl.
  • the 5-15 membered heteroaryl group is a 5-15 membered heteroaryl group having 1-2 heteroatoms independently arbitrarily selected from N, S, or O.
  • the 5-10 membered heteroaryl group is a 5-10 membered heteroaryl group having 1-2 heteroatoms independently arbitrarily selected from N, S, or O.
  • fused heterocycle refers to a cyclic hydrocarbon in which 2-3 rings share two adjacent (ortho) atoms; at least one ring is an aromatic ring containing 1 to 3 heteroatoms selected from O, N and S , in some instances, is a cyclic hydrocarbon in which 2 rings share two adjacent (ortho) atoms; one ring is an aromatic ring containing 1 to 3 heteroatoms selected from O, N, and S, and the other ring is a saturated heterocycle.
  • the 8-15 membered fused heteroaryl group of the present invention has an 8-15 membered fused heterocycle with 1-2 heteroatoms selected from N or O.
  • compositions of the present invention refer to salts of the compounds of the present invention, such salts are safe and effective when used in mammals, and have due biological activity.
  • the compounds of the present invention may contain one or more asymmetric centers and thus occur as racemates, racemic mixtures, single enantiomers, diastereomeric compounds and single diastereomers.
  • the R and S configurations for each asymmetric center the Z and E double bond isomers and the Z and E conformers. Accordingly, single stereoisomers as well as enantiomeric, diastereomeric and geometric (or conformational) mixtures of the compounds of the present invention are within the scope of the present invention. Unless otherwise stated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.
  • treating or “treatment” as referred to throughout this document is conventional, eg, to manage or care for an individual for the purpose of counteracting, alleviating, reducing, alleviating, ameliorating the condition of a disease or disorder .
  • the term "individual” or “patient” includes organisms, such as humans and non-human animals, capable of suffering from a variety of diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport, or that would benefit from administration of the compounds of the present invention .
  • Preferred humans include human patients suffering from or susceptible to suffering from various diseases, disorders or conditions or related states associated with inappropriate nuclear transport-triggered abnormal cellular responses as described herein.
  • non-human animal includes vertebrates, such as mammals, such as non-human primates, sheep, cows, dogs, cats, and rodents (eg, mice), as well as non-mammals, such as chickens, amphibians , reptiles, etc.
  • the compounds of the present invention can be used to treat various diseases, disorders or conditions related to abnormal cellular responses triggered by inappropriate nuclear transport.
  • the study of nuclear transport regulation in biological and pathological phenomena such as the study of kinase-mediated intracellular signal transduction pathways, and the comparative evaluation of new nuclear transport regulators are also valuable.
  • the compounds of the present invention have better cellular activity and selectivity, better water solubility, reduced blood-brain barrier penetration ability to reduce central nervous system toxicity, and reduced hERG activity to reduce cardiotoxicity. More specifically, the compounds of the present invention have the characteristics of high activity, good selectivity and low toxicity, so the compounds of the present invention have better druggability.
  • the synthetic route is as follows:
  • reaction mixture was stirred at 85°C for 10 h under nitrogen protection, then filtered and concentrated under reduced pressure. Separation and purification were carried out by prep-TLC (eluent: ethyl acetate) and prep-HPLC to obtain compound P-11 (22.4 mg, purity 97.67%, yield 17.12%) as a white solid.
  • the synthetic route is as follows:
  • 3-Bromo-1,2,4-triazole (SM1, 4.44 g, 30.0 mmol) was dissolved in 30 ml of DMF and triethylenediamine (6.73 g, 60.0 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. Then (2Z)-isopropyl-3-iodoprop-2-enoate (7.92 g, 33.0 mmol) was added and the final mixture was stirred at room temperature for 1 h. The mixture was poured into 100ml ice water and the aqueous phase was extracted with ethyl acetate (3x50ml).
  • the crude product was purified by preparative TLC, and purified by preparative HPLC (mobile phase: ACN/H2O: 60/40 ⁇ 40/60) to obtain the target product P-25 (12 mg, yield 18.3%) as a white solid.
  • the crude product was purified by preparative TLC, and purified by preparative HPLC (mobile phase: ACN/H2O: 60/40 ⁇ 20/80) to obtain the target product P-26 (16.3 mg, yield: 32.8%) in white solid.
  • Test compounds against different cell lines MM1S (myeloma cells), SKM-1 (human myelodysplastic syndrome cells), LNcap (prostate cancer cells), MOLT-4 (acute lymphoblastic leukemia cells), COLO205 (human colon cancer cells) Cells), RPMI 8226 (human multiple myeloma peripheral blood B lymphocytes) and other proliferation inhibitory activities.
  • MM1S myeloma cells
  • SKM-1 human myelodysplastic syndrome cells
  • LNcap prostate cancer cells
  • MOLT-4 acute lymphoblastic leukemia cells
  • COLO205 human colon cancer cells
  • RPMI 8226 human multiple myeloma peripheral blood B lymphocytes
  • KPT-8602 is compound 124 in the patent (CN105339358A), and its structural formula is as follows:
  • U2OS-Rev(1.4)-NES-mGEP C#52 cell line human osteosarcoma cells labeled with HIV Rev response element with green fluorescent protein
  • U2OS-Rev(1.4)-NES-mGEP C#52 cell line human osteosarcoma cells labeled with HIV Rev response element with green fluorescent protein
  • cells were resuspended in inoculation medium and incubated overnight at 37°C with 5% CO2.
  • Compounds were serially diluted 3-fold to 0.51 nM in DMSO, added to cell culture plates at a 100-fold dilution with medium McCoy'5A, and incubated for 4 hr.
  • PFA paraformaldehyde solution
  • HEK293 cells expressing hERG channels Using HEK293 cells expressing hERG channels, the effect of compounds of the present invention on hERG ion channels was investigated. Compounds were perfused using a perfusion system utilizing its own gravity. Five concentrations of each compound were tested, 0.3, 1, 3, 10, 30 ⁇ M, and at least two cells were tested for each concentration. After the current was stable (or 5 minutes), the current magnitude changes before and after the compound was used were compared to calculate the blocking effect of the compound. The cells were transferred to a perfusion tank and perfused with extracellular fluid. Electrodes were drawn with PC-10 (Narishige, Japan). Whole-cell patch-clamp recordings, with noise filtered at one-fifth of the sampling frequency.
  • hERG tail currents are pure hERG currents.
  • the maximum current induced by the second square wave was detected.
  • the test compound was perfused.
  • the blocking intensity was calculated. The value of the current before the compound was used was denoted as a, and the value when the compound was perfused until the current was stable was denoted as b.
  • the average blocking rate was the average value of the parallel detection data, and the compound concentration and the average blocking rate were fitted with origin software to obtain the IC 50 value.
  • Both the test substance and KPT8602 were orally administered to rats at 10 mg/kg, and the administration volume was 10 mL/kg.
  • test substance solution accurately weigh 10 mg of test substance, add 1 mL DMSO solution, vortex for 5 min, add 0.8 mL Solutol solution, vortex for 5 min, add 1.5 mL PEG400 solution, and mix well , add water to a certain volume of 10mL.
  • Example 46 In vivo efficacy test of the compound in animals
  • MM.1S cells were routinely cultured in RPMI1640 medium containing 10% fetal bovine serum under the culture condition of 5% CO 2 at 37°C; according to cell growth conditions, passage 1 to 2 times a week, and the passage ratio was 1:2 ⁇ 3.
  • the patients were randomly divided into groups according to the tumor volume.
  • control group was given KPT-8602 at a dose of 5 mg/kg, and the experimental group was administered at a dose of 2.5, 5, 7.5, and 10, once a day, and the administration period was 4 weeks.
  • the tumor inhibitory efficacy of the compounds was evaluated by tumor volume tumor inhibition rate (GI) or relative tumor proliferation rate T/C (%).
  • T/C (%) (T RTV /C RTV ) ⁇ 100%, where T RTV represents the relative tumor volume of the treatment group, and C RTV represents the solvent control group relative tumor volume.
  • Example 47 Subacute toxicity test of the compounds of the present invention
  • Oral gavage was administered to SD rats at 3, 9, and 18 mg/kg, with an administration volume of 5 mL/kg, 5 days a week for 2 consecutive weeks.
  • Solvent Weigh an appropriate amount of Solutol and MC to prepare 20% Solutol+0.5% MC.
  • the preparation method is as follows: Weigh an appropriate amount of the sample powder of the test substance, grind, add an appropriate amount of solvent, vortex to completely dissolve the sample and mix well, ultrasonically , prepared into the desired suspension, now with the current use.
  • the observation by the cage includes the appearance of the animal, coat, physical signs, behavioral activities, respiratory status, gland secretion, animal posture, feces, death, etc.
  • Body weight Measure body weight once a week
  • Animal anesthesia and dissection After the administration, the animals were euthanized, and the tissues and organs were visually inspected for abnormalities, such as heart, liver, spleen, lung, kidney, brain, adrenal gland, thymus, testis, epididymis, uterus, and ovary. Calculate the organ coefficient.
  • Organ coefficient [Organ weight (g) / Body weight (g)] * 100

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Abstract

Provided are a 1,2,4-triazole derivative and a preparation method therefor and a use thereof. The 1,2,4-triazole derivative is a compound represented by formula I, or an optical isomer, an enantiomer, a diastereomer, a racemate, or a racemic mixture thereof, or a solvate and a prodrug thereof, or a pharmaceutically acceptable salt thereof. The compound can be used for treating various diseases, disorders or conditions related to abnormal cellular responses triggered by inappropriate nuclear transport.

Description

1,2,4-三氮唑衍生物及其制备方法和用途1,2,4-Triazole derivatives and their preparation and use 技术领域technical field
本发明属于医药技术领域，具体涉及新的化合物、包含该化合物的药物组合、制备该化合物的方法和该化合物在治疗多种与不当的核转运触发的异常细胞应答相关的疾病、紊乱或病症的用途。The present invention belongs to the technical field of medicine, and particularly relates to novel compounds, pharmaceutical combinations comprising the compounds, methods for preparing the compounds, and the use of the compounds in the treatment of various diseases, disorders or conditions related to abnormal cellular responses triggered by inappropriate nuclear transport. use.
背景技术Background technique
真核生物的细胞核具有双层膜结构，大分子物质需要通过核质转运受体的介导才能进行跨核膜转运。特异的蛋白和RNA由特异的转运分子转运入和转运出细胞核，其中将分子转运入细胞核的，则这被分类为核输入蛋白，若它们将分子输出细胞核，则被分类为核输出蛋白。其中核输出蛋白1(exportin 1,XPO1)，也称作染色体区域稳定蛋白1(CRM1)是高尔基体的组成部分，同时也是核质转运的重要转运受体之一，是主要的输出蛋白核受体。其在若干种肿瘤细胞(例如：人类的卵巢癌、宫颈癌、胰腺癌、肝细胞癌、白血病、骨髓瘤)中高度表达，现已成为抗肿瘤药研发的重要靶标。The nucleus of eukaryotes has a double-layered membrane structure, and macromolecules need to be transported across the nuclear membrane through the mediation of nucleocytoplasmic transport receptors. Specific proteins and RNAs are transported into and out of the nucleus by specific transport molecules, wherein those that transport the molecules into the nucleus are classified as nuclear import proteins, and those that transport molecules out of the nucleus are classified as nuclear export proteins. Among them, nuclear export protein 1 (exportin 1, XPO1), also known as chromosomal region stabilizing protein 1 (CRM1), is a component of the Golgi apparatus and one of the important transport receptors for nucleocytoplasmic transport. body. It is highly expressed in several tumor cells (eg, human ovarian cancer, cervical cancer, pancreatic cancer, hepatocellular carcinoma, leukemia, myeloma), and has now become an important target for the development of antitumor drugs.
XPO1可以介导200余种的蛋白进行转运，包括抑癌基因蛋白(tumor suppressor proteins，TSPs)、P53、P73等，抗凋亡蛋白(IAPs)、核仁磷酸蛋白(nucleophsmim,NPM)、存活素以及生长调节蛋白(GRPs)P21和P27等。通过抑制XPO1介导出核转运，能够阻断肿瘤抑制蛋白和/或生长调控因子的核输出。XPO1抑制因子可诱导癌细胞的凋亡，甚至是在致癌激活信号或生长激励信号存在的情况下，而不影响正常的(未被转化的)细胞。癌症细胞中XPO1表达水平平均高出正常细胞2-4倍，这使得TSPs、IAPs、GRPs大部分被运出核，对于肿瘤抑制蛋白识别和诱导癌细胞凋亡起到不利作用，使得癌细胞增殖分化不受限制。XPO1 can mediate the transport of more than 200 kinds of proteins, including tumor suppressor proteins (TSPs), P53, P73, etc., anti-apoptotic proteins (IAPs), nucleophsmim (NPM), survivin And growth regulatory proteins (GRPs) P21 and P27. By inhibiting XPO1-mediated nuclear transport, the nuclear export of tumor suppressor proteins and/or growth regulators can be blocked. XPO1 inhibitors can induce apoptosis in cancer cells, even in the presence of oncogenic activating signals or growth stimulating signals, without affecting normal (untransformed) cells. The expression level of XPO1 in cancer cells is 2-4 times higher than that in normal cells on average, which makes most of TSPs, IAPs, and GRPs transported out of the nucleus, which has an adverse effect on tumor suppressor protein recognition and induction of cancer cell apoptosis, which makes cancer cells proliferate. Differentiation is not limited.
第一代的XPO1抑制剂selinexor在临床前及临床研究中表现出高的血脑屏障渗透和较大毒性问题，亟需新一代XPO1抑制剂满足临床需求。The first-generation XPO1 inhibitor selinexor showed high blood-brain barrier penetration and greater toxicity in preclinical and clinical studies, and a new generation of XPO1 inhibitors is urgently needed to meet clinical needs.
发明内容SUMMARY OF THE INVENTION
为解决现有技术中存在的问题，本发明提供一种1,2,4-三氮唑衍生物及其制备方法和用途。In order to solve the problems existing in the prior art, the present invention provides a 1,2,4-triazole derivative and a preparation method and application thereof.
为达上述技术效果，本发明采用以下技术方案：In order to reach above-mentioned technical effect, the present invention adopts following technical scheme:
本发明目的之一在于提供一种化合物，其特征在于，所述化合物为式I所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐；One of the objects of the present invention is to provide a compound, characterized in that the compound is the compound represented by formula I, or its optical isomer, enantiomer, diastereomer, racemate or racemate a mixture, or a solvate, prodrug, or a pharmaceutically acceptable salt thereof;
Figure PCTCN2021127847-appb-000001
Figure PCTCN2021127847-appb-000001
其中：in:
R 2、R 3各自独立的选自H或C 1～C 3的烷基； R 2 and R 3 are each independently selected from H or C 1 -C 3 alkyl groups;
R 4选自H或C 1～C 3的烷基； R 4 is selected from H or C 1 -C 3 alkyl;
R 1选自未取代或被A任意取代的以下基团：C 6-C 15芳基、5-15元杂芳基、C 6-C 15环烷基或5-15元杂环烷基；取代基A选自：卤素、羟基、羰基(＝O)、-CN、C 1-C 6烷基，C 1-C 6烷氧基、羟基C 1-C 6烷基、卤代C 1-C 6烷基、氰基C 1-C 6烷基、或者未取代或被B任意取代的以下集团：C 6-C 10芳基、5-10元杂芳基、C 6-C 10环烷基、5-10元杂环烷基或
Figure PCTCN2021127847-appb-000002
R 5选自H或C 1～C 3的烷基、R 6选自H、C 1-C 6烷基、卤代C 1-C 6烷基、氰基C 1-C 6烷基、C 6-C 10芳基、卤代C 6-C 10芳基、C 1-C 3烷基取代的C 6-C 10芳基；取代基B选自：卤素、羟基、羰基(＝O)、-CN、C 1-C 6烷基、C 1-C 6烷氧基、羟基C 1-C 6烷基、卤代C 1-C 6烷基、氰基C 1-C 6烷基。 R 1 is selected from the following groups unsubstituted or optionally substituted by A: C 6 -C 15 aryl, 5-15 membered heteroaryl, C 6 -C 15 cycloalkyl or 5-15 membered heterocycloalkyl; Substituent A is selected from: halogen, hydroxy, carbonyl (=O), -CN, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, hydroxy C 1 -C 6 alkyl, halogenated C 1 - C 6 alkyl, cyano C 1 -C 6 alkyl, or the following groups unsubstituted or optionally substituted by B: C 6 -C 10 aryl, 5-10 membered heteroaryl, C 6 -C 10 cycloalkane base, 5-10 membered heterocycloalkyl or
Figure PCTCN2021127847-appb-000002
R 5 is selected from H or C 1 -C 3 alkyl, R 6 is selected from H, C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, cyano C 1 -C 6 alkyl, C 6 -C 10 aryl, halogenated C 6 -C 10 aryl, C 1 -C 3 alkyl substituted C 6 -C 10 aryl; Substituent B is selected from: halogen, hydroxyl, carbonyl (=O), -CN, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, hydroxy C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, cyano C 1 -C 6 alkyl.
在一些具体的实施例中，R 2、R 3各自独立的选自H、甲基或乙基。 In some specific embodiments, R 2 , R 3 are each independently selected from H, methyl or ethyl.
在一些具体的实施例中，R 4选自H、甲基或乙基。 In some specific embodiments, R4 is selected from H, methyl or ethyl.
在一些具体的实施例中，R1选自未取代或任意取代的C5-C10芳基；取代基选自卤素、羟基、羰基(＝O)、-CN、C 1-C 3烷基，C 1-C 3烷氧基、羟基C 1-C 3烷基、卤代C 1-C 3烷基、氰基C 1-C 3烷基、C 6-C 10芳基、5-10元杂芳基、C 6-C 10环烷基、5-10元杂环烷。 In some specific embodiments, R1 is selected from unsubstituted or optionally substituted C5-C10 aryl; substituents are selected from halogen, hydroxyl, carbonyl (=O), -CN, C 1 -C 3 alkyl, C 1 -C 3 alkoxy, hydroxy C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl, cyano C 1 -C 3 alkyl, C 6 -C 10 aryl, 5-10 membered heteroaryl base, C 6 -C 10 cycloalkyl, 5-10 membered heterocycloalkane.
在一些具体的实施例中，R1选自未取代或被A任意取代的5-6元杂芳基；取代基A选自：卤素、羟基、羰基(＝O)、-CN、C 1-C 3烷基，C 1-C 3烷氧基、羟基C 1-C 3烷基、卤代C 1-C 3烷基、氰基C 1-C 3烷基、或者未取代或被B任意取代的以下基团：C 6-C 10芳基、5-10元杂芳基、C 6-C 10环烷基、5-10元杂环烷基或
Figure PCTCN2021127847-appb-000003
R 5选自H或C 1～C 3的烷基、R 6选自H、C 1-C 3烷基、卤代C 1-C 3烷基、氰基C 1-C 3烷基、C 6-C 10芳基、卤代C 6-C 10芳基、C 1-C 3烷基取代的C 6-C 10芳基；取代基B选自：卤素、羟基、羰基(＝O)、-CN、C 1-C 3烷基、C 1-C 3烷氧基、羟基C 1-C 3烷基、卤代C 1-C 3烷基、氰基 C 1-C 3烷基。 In some specific embodiments, R1 is selected from unsubstituted or optionally substituted by A 5-6-membered heteroaryl; Substituent A is selected from: halogen, hydroxyl, carbonyl (=O), -CN, C 1 -C 3 alkyl, C 1 -C 3 alkoxy, hydroxy C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl, cyano C 1 -C 3 alkyl, or unsubstituted or optionally substituted by B of the following groups: C 6 -C 10 aryl, 5-10 membered heteroaryl, C 6 -C 10 cycloalkyl, 5-10 membered heterocycloalkyl or
Figure PCTCN2021127847-appb-000003
R 5 is selected from H or C 1 -C 3 alkyl, R 6 is selected from H, C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl, cyano C 1 -C 3 alkyl, C 6 -C 10 aryl, halogenated C 6 -C 10 aryl, C 1 -C 3 alkyl substituted C 6 -C 10 aryl; Substituent B is selected from: halogen, hydroxyl, carbonyl (=O), -CN, C 1 -C 3 alkyl, C 1 -C 3 alkoxy, hydroxy C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl, cyano C 1 -C 3 alkyl.
在一些具体的实施例中，R1选自未取代或任意取代的8-15元稠杂芳基；取代基选自卤素、羟基、-CN、C 1-C 3烷基，C 1-C 3烷氧基、羟基C 1-C 3烷基、卤代C 1-C 3烷基。 In some specific embodiments, R1 is selected from unsubstituted or optionally substituted 8-15-membered fused heteroaryl; substituents are selected from halogen, hydroxyl, -CN, C 1 -C 3 alkyl, C 1 -C 3 Alkoxy, hydroxy C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl.
在一些具体的实施例中，R1选自未取代或被任意取代的吡咯基、呋喃基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基、三唑基、噻二唑基或噁二唑基。取代基可以如前任一所述。In some specific embodiments, R1 is selected from unsubstituted or optionally substituted pyrrolyl, furyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl , thiadiazolyl or oxadiazolyl. Substituents may be as described above.
R1是具有1、2或3个独立地选自下组的杂原子的任意取代的6元杂芳基，该组由以下各项组成：氮、氧和硫。R1 is an optionally substituted 6-membered heteroaryl having 1, 2 or 3 heteroatoms independently selected from the group consisting of nitrogen, oxygen and sulfur.
在一些具体的实施例中，R1选自未取代或被任意取代的吡啶基、嘧啶基、吡嗪基、哒嗪基或三嗪基。取代基可以如前任一所述。In some specific embodiments, R1 is selected from unsubstituted or optionally substituted pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, or triazinyl. Substituents may be as described above.
在本发明的一些实施例中，本发明所述的化合物，所述化合物为式I-1所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐；In some embodiments of the present invention, the compound of the present invention is the compound represented by formula I-1, or its optical isomer, enantiomer, diastereomer, racemate or a racemic mixture, or a solvate, prodrug, or a pharmaceutically acceptable salt thereof;
Figure PCTCN2021127847-appb-000004
Figure PCTCN2021127847-appb-000004
其中，R1如前所述。Wherein, R1 is as described above.
在一些更具体的实施例中，化合物选自如下：In some more specific embodiments, the compound is selected from the group consisting of:
Figure PCTCN2021127847-appb-000005
Figure PCTCN2021127847-appb-000005
Figure PCTCN2021127847-appb-000006
Figure PCTCN2021127847-appb-000006
Figure PCTCN2021127847-appb-000007
Figure PCTCN2021127847-appb-000007
Figure PCTCN2021127847-appb-000008
Figure PCTCN2021127847-appb-000008
Figure PCTCN2021127847-appb-000009
Figure PCTCN2021127847-appb-000009
Figure PCTCN2021127847-appb-000010
Figure PCTCN2021127847-appb-000010
Figure PCTCN2021127847-appb-000011
Figure PCTCN2021127847-appb-000011
Figure PCTCN2021127847-appb-000012
Figure PCTCN2021127847-appb-000012
本发明还提供一种化合物，其特征在于，为式II所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药或其药学上可接受的盐；The present invention also provides a compound, which is characterized in that it is a compound represented by formula II, or an optical isomer, enantiomer, diastereomer, racemate or racemic mixture thereof, or a solvate thereof drug, prodrug or a pharmaceutically acceptable salt thereof;
Figure PCTCN2021127847-appb-000013
Figure PCTCN2021127847-appb-000013
其中：环C选自5-6元杂芳基、5-6元杂环基，其中所述杂芳基或杂环基任选进一步被一个或多个R 7取代；优选的，环C选自5-6元杂环基，其中所述杂环基任选进一步被一个或两个R 7取代； Wherein: ring C is selected from 5-6 membered heteroaryl, 5-6 membered heterocyclyl, wherein said heteroaryl or heterocyclyl is optionally further substituted by one or more R 7 ; preferably, ring C is selected from from 5-6 membered heterocyclyl, wherein said heterocyclyl is optionally further substituted with one or two R7 ;
R 7选自氢原子、卤素、氰基、羟基、C 1～C 8烷基、C 3～C 10环烷基、C 1～C 8烷氧基、C 1～C 8卤代烷基、C 3～C 10卤代环烷基、C 1～C 8卤代烷氧基；优选的，R 7选自氢原子、卤素、氰基、羟基、C 1～C 3烷基、C 3～C 6环烷基、C 1～C 3烷氧基、C 1～C 3卤代烷基、C 3～C 6卤代环烷基、C 1～C 3卤代烷氧基； R 7 is selected from hydrogen atom, halogen, cyano, hydroxyl, C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, C 1 -C 8 alkoxy, C 1 -C 8 haloalkyl, C 3 ~C 10 halogenated cycloalkyl, C 1 -C 8 halogenated alkoxy; preferably, R 7 is selected from hydrogen atom, halogen, cyano, hydroxyl, C 1 -C 3 alkyl, C 3 -C 6 cycloalkane base, C 1 -C 3 alkoxy, C 1 -C 3 haloalkyl, C 3 -C 6 halocycloalkyl, C 1 -C 3 haloalkoxy;
R 8选自H、或者未取代或被一个或多个D任意取代的以下基团：C 6～C 10芳基、C 2～C 10杂芳基、C 3～C 10杂环基、C 10～C 20稠环化合物；取代基D选自：氢原子、卤素、羟基、羰基(＝O)、氰基、C 1～C 8烷基，氨基、C 1～C 8烷基取代氨基、羟基取代的C 1～C 8烷基、C 1～C 8卤代烷基、氰基取代的C 1～C 8烷基；优选的，R 8选自H、或者未取代或被一个或多个D任意取代的以下基团：苯、吡咯、呋喃、噻吩、咪唑、噁唑、噻唑、吡唑、嘧啶、喹啉、异喹啉、嘌呤、四唑、三嗪和咔唑或萘；取代基D选自：氢原子、卤素、羟基、羰基(＝O)、氰基、C 1～C 3烷基，氨基、C 1～C 3烷基取代氨基、羟基取代的C 1～C 3烷基、C 1～C 3卤代烷基、氰基取代的C 1～C 3烷基； R 8 is selected from H, or the following groups unsubstituted or optionally substituted by one or more D: C 6 -C 10 aryl, C 2 -C 10 heteroaryl, C 3 -C 10 heterocyclyl, C 10 - C20 fused ring compound; Substituent D is selected from: hydrogen atom, halogen, hydroxyl, carbonyl (=O), cyano, C1 - C8 alkyl, amino, C1 - C8 alkyl substituted amino, Hydroxy-substituted C 1 -C 8 alkyl, C 1 -C 8 haloalkyl, cyano-substituted C 1 -C 8 alkyl; preferably, R 8 is selected from H, or unsubstituted or by one or more D Optionally substituted: benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyrimidine, quinoline, isoquinoline, purine, tetrazole, triazine and carbazole or naphthalene; Substituent D Selected from: hydrogen atom, halogen, hydroxyl, carbonyl (=O), cyano group, C 1 -C 3 alkyl, amino, C 1 -C 3 alkyl substituted amino, hydroxyl substituted C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, cyano-substituted C 1 -C 3 alkyl;
R 9选自未取代或被一个或多个E任意取代的以下基团：羟基、氨基；取代基E选自：卤素、羟基、C 1～C 6烷基，C 3～C 10环烷基、C 2～C 10杂环基、C 1～C 6烷氧基、C 1～C 6卤代烷基、C 3～C 10卤代环烷基、
Figure PCTCN2021127847-appb-000014
或与被取代的原子形成4～8元杂环；R 9选自：氢原子、羟基、C 1～C 6烷基，C 3～C 10环烷基、C 1～C 6烷氧基、C 1～C 6卤代烷基、C 3～C 10卤代环烷基、C 6～C 10芳基、C 2～C 10杂芳基或C 2～C 10杂环基。 R 9 is selected from the following groups unsubstituted or optionally substituted by one or more E: hydroxyl, amino; Substituent E is selected from: halogen, hydroxyl, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl , C 2 -C 10 heterocyclyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 3 -C 10 halocycloalkyl,
Figure PCTCN2021127847-appb-000014
Or form a 4-8 membered heterocycle with the substituted atom; R 9 is selected from: hydrogen atom, hydroxyl, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 3 -C 10 halocycloalkyl, C 6 -C 10 aryl, C 2 -C 10 heteroaryl or C 2 -C 10 heterocyclic.
R 9选自未取代或被一个或多个E任意取代的以下基团：羟基、氨基；取代基E选自：卤素、羟基、C 1～C 4烷基，C 3～C 8环烷基、C 2～C 8杂环基、C 1～C 4烷氧基、C 1～C 4卤代烷基、C 3～C 8卤代环烷基、
Figure PCTCN2021127847-appb-000015
或与被取代的原子形成4～6元杂环；R 9选自：氢原子、羟基、C 1～C 3烷基，C 3～C 8环烷基、C 1～C 3烷 R 9 is selected from the following groups unsubstituted or optionally substituted by one or more E: hydroxyl, amino; Substituent E is selected from: halogen, hydroxyl, C 1 -C 4 alkyl, C 3 -C 8 cycloalkyl , C 2 -C 8 heterocyclyl, C 1 -C 4 alkoxy, C 1 -C 4 haloalkyl, C 3 -C 8 halocycloalkyl,
Figure PCTCN2021127847-appb-000015
Or form a 4-6 membered heterocycle with the substituted atom; R 9 is selected from: hydrogen atom, hydroxyl, C 1 -C 3 alkyl, C 3 -C 8 cycloalkyl, C 1 -C 3 alkane
氧基、C 1～C 3卤代烷基、C 3～C 8卤代环烷基、C 6～C 10芳基、C 2～C 8杂芳基或C 2～C 8杂环基。 Oxy group, C 1 -C 3 haloalkyl group, C 3 -C 8 halocycloalkyl group, C 6 -C 10 aryl group, C 2 -C 8 heteroaryl group or C 2 -C 8 heterocyclic group.
在本发明的一些实施例中，本发明还提供如式II-1或式II-2所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药或其药学上可接受的盐：In some embodiments of the present invention, the present invention also provides compounds represented by formula II-1 or formula II-2, or optical isomers, enantiomers, diastereomers, racemates or exoisomers thereof A racemic mixture, or a solvate, prodrug or a pharmaceutically acceptable salt thereof:
Figure PCTCN2021127847-appb-000016
Figure PCTCN2021127847-appb-000016
其中，R 8和R 9如前所述；在一些更具体的实施例中，R 8为H或嘧啶。 wherein R 8 and R 9 are as previously described; in some more specific embodiments, R 8 is H or pyrimidine.
在一些更具体的实施例中，本发明还提供如下所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药或其药学上可接受的盐：In some more specific embodiments, the present invention also provides compounds shown below, or optical isomers, enantiomers, diastereomers, racemates or racemic mixtures thereof, or solvates thereof , a prodrug or a pharmaceutically acceptable salt thereof:
Figure PCTCN2021127847-appb-000017
Figure PCTCN2021127847-appb-000017
Figure PCTCN2021127847-appb-000018
Figure PCTCN2021127847-appb-000018
Figure PCTCN2021127847-appb-000019
Figure PCTCN2021127847-appb-000019
Figure PCTCN2021127847-appb-000020
Figure PCTCN2021127847-appb-000020
本发明目的之二在于提供所述化合物的制备方法。The second object of the present invention is to provide a preparation method of the compound.
在本发明的一种实施例中，提供式I-1所示化合物的制备方法：In one embodiment of the present invention, the preparation method of the compound shown in formula I-1 is provided:
Figure PCTCN2021127847-appb-000021
Figure PCTCN2021127847-appb-000021
本文目的之三在于提供一种药物组合物，其包括式I和式II所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐，以及药学上可接受的载体。The third purpose of this paper is to provide a pharmaceutical composition comprising the compounds represented by formula I and formula II, or optical isomers, enantiomers, diastereomers, racemates or racemic mixtures thereof, or a solvate, prodrug thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
本文目的之四在于提供式I和式II所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐、或本发明所述的组合物作为XPO1抑制剂的应用。The fourth purpose of this paper is to provide compounds represented by formula I and formula II, or optical isomers, enantiomers, diastereomers, racemates or racemic mixtures thereof, or solvates and prodrugs thereof , or a pharmaceutically acceptable salt thereof, or the application of the composition of the present invention as an XPO1 inhibitor.
本文目的之五在于提供式I和式II所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐、或本发明所述的组合物作为制备治疗多种与不当的核转运触发的异常细胞应答相关的疾病、紊乱或病症的药物中的用途。The fifth purpose of this paper is to provide compounds represented by formula I and formula II, or optical isomers, enantiomers, diastereomers, racemates or racemic mixtures thereof, or solvates and prodrugs thereof , or a pharmaceutically acceptable salt thereof, or the composition of the present invention as a use in the manufacture of a medicament for the treatment of various diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport.
本发明式I和式II所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐或本发明所述的组合物对于治疗多种不当的核转运触发的异常细胞应答相关的疾病、紊乱或病症也是有用的。因此本发明的一个实施例是本发明 的化合物或其药学上可接受的盐用于治疗多种与不当的核转运触发的异常细胞应答的疾病、紊乱或病症的用途。本发明提供的化合物对于生物学与病理学现象中的核转运调节的研究，例如激酶介导的细胞内信号转到通路的研究以及新的核转运调节剂的比较评价也是有用的。The compounds represented by formula I and formula II of the present invention, or optical isomers, enantiomers, diastereomers, racemates or racemic mixtures thereof, or solvates, prodrugs, or pharmaceuticals thereof The acceptable salts above or the compositions described herein are also useful for the treatment of various diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport. Thus one embodiment of the present invention is the use of a compound of the present invention, or a pharmaceutically acceptable salt thereof, for the treatment of various diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport. The compounds provided by the present invention are also useful for the study of nuclear transport regulation in biological and pathological phenomena, such as the study of kinase-mediated intracellular signal transduction pathways and the comparative evaluation of novel nuclear transport modulators.
在一些实施例中，本发明所述的多种与不当的核转运触发的异常细胞应答相关的疾病、紊乱或病症包括但不限于增生性紊乱、癌症、炎症性紊乱、自身免疫性紊乱、病毒感染、神经退行性紊乱、眼科紊乱、异常组织生长紊乱、与食物摄取有关的紊乱、过敏以及呼吸紊乱。In some embodiments, various diseases, disorders or conditions described herein that are associated with abnormal cellular responses triggered by inappropriate nuclear transport include, but are not limited to, proliferative disorders, cancers, inflammatory disorders, autoimmune disorders, viruses Infections, neurodegenerative disorders, ophthalmic disorders, disorders of abnormal tissue growth, disorders related to food intake, allergies, and respiratory disorders.
本文还提供一种用于患有治疗多种不当的核转运触发的异常细胞应答相关的疾病、紊乱或病症的患者的方法，其包括向患者施用治疗有效量的如本文所定义的式I所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐或其药物组合物。Also provided herein is a method for treating a patient having a disease, disorder or condition associated with various inappropriate nuclear transport-triggered abnormal cellular responses, comprising administering to the patient a therapeutically effective amount of formula I as defined herein The compound shown, or its optical isomer, enantiomer, diastereomer, racemate or racemic mixture, or its solvate, prodrug, or its pharmaceutically acceptable salt or its drug combination.
在一些实施例中，患者是人类。In some embodiments, the patient is a human.
本发明术语若无特别说明定义如下：Unless otherwise specified, the terms of the present invention are defined as follows:
术语“卤素”意指-F(在本文中有时被称作“氟”)、-Cl、-Br和-I。The term "halogen" means -F (sometimes referred to herein as "fluoro"), -Cl, -Br and -I.
术语“C1-C3烷基”、“C1-C6烷基”、“C2-C6烷基”和“C3-C6烷基”是指分别具有一个到三个、一个到六个、两个到六个或三个到六个碳原子的饱和的直链或分支链一价烃基。实例包含(但不限于)甲基、乙基、1-丙基、异丙基、1-丁基、异丁基、仲丁基、叔丁基、2-甲基-2-丙基、戊基、新戊基和己基。The terms "C1-C3 alkyl", "C1-C6 alkyl", "C2-C6 alkyl" and "C3-C6 alkyl" refer to groups having one to three, one to six, two to six, respectively A saturated linear or branched monovalent hydrocarbon radical of one or three to six carbon atoms. Examples include, but are not limited to, methyl, ethyl, 1-propyl, isopropyl, 1-butyl, isobutyl, sec-butyl, tert-butyl, 2-methyl-2-propyl, pentyl base, neopentyl and hexyl.
术语“C1-C6烷氧基”是指具有一到六个碳原子的饱和直链或分支链一价烷氧基，其中键是在氧原子上。实例包含甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基和叔丁氧基。The term "C1-C6 alkoxy" refers to a saturated straight or branched monovalent alkoxy group having one to six carbon atoms, wherein the bond is on an oxygen atom. Examples include methoxy, ethoxy, propoxy, isopropoxy, butoxy and tert-butoxy.
术语“羟基C1-C6烷基-”是指具有一个到六个碳原子的饱和直链或分支链一价烷基，其中至少一个碳原子被羟基取代。The term "hydroxy C1-C6 alkyl-" refers to a saturated straight or branched chain monovalent alkyl group having one to six carbon atoms, wherein at least one carbon atom is substituted with a hydroxyl group.
术语“卤代C1-C6烷基-”是指具有一个到六个碳原子的饱和直链或分支链一价烷基，其中至少一个碳原子被卤素取代。The term "haloC1-C6alkyl-" refers to a saturated straight or branched monovalent alkyl group having one to six carbon atoms, wherein at least one carbon atom is substituted with a halogen.
术语“氰基C1-C6烷基-”是指具有一个到六个碳原子的饱和直链或分支链一价烷基，其中一个碳原子被氰基取代。The term "cyano C1-C6 alkyl-" refers to a saturated straight or branched monovalent alkyl group having one to six carbon atoms, one of which is replaced by a cyano group.
术语“C3-C6环烷基”是指环丙基、环丁基、环戊基或环己基。The term "C3-C6 cycloalkyl" refers to cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
术语“杂环”是指除了含有碳原子以外还含有1-4个选自由氧原子、硫原子及氮原子组成的组中的杂原子的单环式或双环式的非芳香族杂环，作为具体例，可以举出：氮杂环丁烷、吡咯烷、吡唑烷(pyrazolidine)、哌啶、氧杂环丁烷、四氢呋喃、四氢吡喃、四氢噻吩、二氢咪唑、咪唑烷 (imidazolidine)、四氢吡嗪、哌嗪、吗啉等除了含有碳原子以外还含有1个或2个选自由氧原子、硫原子及氮原子组成的组中的杂原子的4-7元单环式非芳香族杂环；氮杂双环[3.1.0]己烷等除了含有碳原子以外还含有1-4个选自由氧原子、硫原子及氮原子组成的组中的杂原子的6-8元双环式非芳香族杂环。在一些实施例中，5-15元杂环烷基是具有独立地任意选自N、S或O的1～2个杂原子的5-15元杂环烷基。在一些实施例中，5-10元杂环烷基是具有独立地任意选自N、S或O的1～2个杂原子的5-10元杂环烷基。The term "heterocycle" refers to a monocyclic or bicyclic non-aromatic heterocycle containing, in addition to carbon atoms, 1-4 heteroatoms selected from the group consisting of oxygen atoms, sulfur atoms and nitrogen atoms, as Specific examples include azetidine, pyrrolidine, pyrazolidine, piperidine, oxetane, tetrahydrofuran, tetrahydropyran, tetrahydrothiophene, dihydroimidazole, imidazolidine ( imidazolidine), tetrahydropyrazine, piperazine, morpholine, etc. 4-7 membered monocyclic ring containing, in addition to carbon atoms, one or two hetero atoms selected from the group consisting of oxygen atoms, sulfur atoms and nitrogen atoms Non-aromatic heterocycles of the formula; azabicyclo[3.1.0]hexane, etc. 6-8 containing 1-4 heteroatoms selected from the group consisting of oxygen atoms, sulfur atoms and nitrogen atoms in addition to carbon atoms A membered bicyclic non-aromatic heterocycle. In some embodiments, the 5-15 membered heterocycloalkyl is a 5-15 membered heterocycloalkyl having 1-2 heteroatoms independently optionally selected from N, S, or O. In some embodiments, the 5-10 membered heterocycloalkyl is a 5-10 membered heterocycloalkyl having 1-2 heteroatoms independently optionally selected from N, S, or O.
术语“杂芳基”代表环中多达3-15个原子的稳定的单环或每个环中多达3-15个原子双环碳环，其中至少一个环为芳香环且含有1-4个选自O、N和S的杂原子。本定义范围内的杂芳基包括但不限于吖啶基、咔唑基、噌啉基、喹噁啉基、吡唑基、吲哚基、苯并三唑基、呋喃基、噻吩基、苯并噻吩基、苯并呋喃基、喹啉基、异喹啉基、噁唑基、异噁唑基、吲哚基、吡嗪基、哒嗪基、吡啶基、嘧啶基、吡咯基。在一些实施例中，5-15元杂芳基是具有独立地任意选自N、S或O的1～2个杂原子的5-15元杂芳基。在一些实施例中，5-10元杂芳基是具有独立地任意选自N、S或O的1～2个杂原子的5-10元杂芳基。The term "heteroaryl" represents a stable monocyclic ring of up to 3-15 atoms in the ring or a bicyclic carbocyclic ring of up to 3-15 atoms in each ring, wherein at least one ring is aromatic and contains 1-4 A heteroatom selected from O, N and S. Heteroaryl groups within the scope of this definition include, but are not limited to, acridinyl, carbazolyl, cinnolinyl, quinoxolinyl, pyrazolyl, indolyl, benzotriazolyl, furyl, thienyl, benzene thienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl. In some embodiments, the 5-15 membered heteroaryl group is a 5-15 membered heteroaryl group having 1-2 heteroatoms independently arbitrarily selected from N, S, or O. In some embodiments, the 5-10 membered heteroaryl group is a 5-10 membered heteroaryl group having 1-2 heteroatoms independently arbitrarily selected from N, S, or O.
术语“稠杂环”是指2-3个环共用两个相邻(邻位)原子的环烃；其中至少一个环为含有1到3个选自O、N和S的杂原子的芳香环，在一些实例中，为2个环共用两个相邻(邻位)原子的环烃；其中一个环为含有1到3个选自O、N和S的杂原子的芳香环，另一个环为饱和杂环。在一些实施例中，本发明8-15元稠杂芳基具有选自N或O的1～2个杂原子的8～15元稠杂环。The term "fused heterocycle" refers to a cyclic hydrocarbon in which 2-3 rings share two adjacent (ortho) atoms; at least one ring is an aromatic ring containing 1 to 3 heteroatoms selected from O, N and S , in some instances, is a cyclic hydrocarbon in which 2 rings share two adjacent (ortho) atoms; one ring is an aromatic ring containing 1 to 3 heteroatoms selected from O, N, and S, and the other ring is a saturated heterocycle. In some embodiments, the 8-15 membered fused heteroaryl group of the present invention has an 8-15 membered fused heterocycle with 1-2 heteroatoms selected from N or O.
本发明的“药学上可接受的盐”是指本发明化合物的盐，这类盐用于哺乳动物体内时具有安全性和有效性，且具有应有的生物活性。The "pharmaceutically acceptable salts" of the present invention refer to salts of the compounds of the present invention, such salts are safe and effective when used in mammals, and have due biological activity.
本发明的化合物可以含有一个或多个不对称中心，并因此以消旋体、外消旋混合物、单一对映体、非对映异构体化合物和单一非对映体的形式出现。如针对每个不对称中心的R和S构型，Z与E双键异构体以及Z和E构象异构体。因此，本发明化合物的单一立体异构体连同对映异构、非对映异构和几何(或构象)混合物都在本发明的范围之内。除非另行说明，本发明的化合物的所有互变异构形式在本发明的范围之内。The compounds of the present invention may contain one or more asymmetric centers and thus occur as racemates, racemic mixtures, single enantiomers, diastereomeric compounds and single diastereomers. As for the R and S configurations for each asymmetric center, the Z and E double bond isomers and the Z and E conformers. Accordingly, single stereoisomers as well as enantiomeric, diastereomeric and geometric (or conformational) mixtures of the compounds of the present invention are within the scope of the present invention. Unless otherwise stated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.
本文件通篇提及的术语“治疗(treating)”或“治疗(treatment)”的使用是常规的，例如为了抵抗、减轻、减少、缓解、改善疾病或病症的病况的目的来管理或护理个体。Use of the terms "treating" or "treatment" as referred to throughout this document is conventional, eg, to manage or care for an individual for the purpose of counteracting, alleviating, reducing, alleviating, ameliorating the condition of a disease or disorder .
术语“个体”或“患者”包括能够患多种与不当的核转运触发的异常细胞应答相关的疾病、紊乱或病症或者能够受益于给药本发明的化合物的有机体，例如人和非人的动物。优选的人包括患有或易于患有如本文所述的多种与不当的核转运触发的异常细胞应答相关的疾病、紊乱或病症或相关 的状态的人患者。术语“非人的动物”包括脊椎动物，例如哺乳动物，如非人灵长类动物、绵羊、母牛、狗、猫和啮齿动物(例如小鼠)，以及非哺乳动物，例如鸡、两栖动物、爬行动物等。The term "individual" or "patient" includes organisms, such as humans and non-human animals, capable of suffering from a variety of diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport, or that would benefit from administration of the compounds of the present invention . Preferred humans include human patients suffering from or susceptible to suffering from various diseases, disorders or conditions or related states associated with inappropriate nuclear transport-triggered abnormal cellular responses as described herein. The term "non-human animal" includes vertebrates, such as mammals, such as non-human primates, sheep, cows, dogs, cats, and rodents (eg, mice), as well as non-mammals, such as chickens, amphibians , reptiles, etc.
有益效果：本发明式所述的化合物可以用于治疗多种不当的核转运触发的异常细胞应答相关的疾病、紊乱或病症。对于生物学与病理学现象中的核转运调节的研究，例如激酶介导的细胞内信号转到通路的研究以及新的核转运调节剂的比较评价也具有研究价值。相对于现有技术，本发明的化合物细胞活性及选择性更好、水溶性更好、血脑屏障穿透能力降低以减少中枢神经毒性、hERG活性降低以减少心脏毒性。更具体地，本发明的化合物具有活性高、选择性好、毒性低的特点，因此本发明的化合物具有更好的成药性。Beneficial effects: The compounds of the present invention can be used to treat various diseases, disorders or conditions related to abnormal cellular responses triggered by inappropriate nuclear transport. The study of nuclear transport regulation in biological and pathological phenomena, such as the study of kinase-mediated intracellular signal transduction pathways, and the comparative evaluation of new nuclear transport regulators are also valuable. Compared with the prior art, the compounds of the present invention have better cellular activity and selectivity, better water solubility, reduced blood-brain barrier penetration ability to reduce central nervous system toxicity, and reduced hERG activity to reduce cardiotoxicity. More specifically, the compounds of the present invention have the characteristics of high activity, good selectivity and low toxicity, so the compounds of the present invention have better druggability.
具体实施方式Detailed ways
以下结合实施例进一步描述本发明，但这些实施例并非限制着本发明的范围，凡在本发明的构思前提下对本发明制备方法的简单改进都属于本发明的保护范围之内。下面实施例未注明具体条件的实验方法，通常按照本领域的公知手段。下述实施例中所用的试验材料，如无特殊说明，均为自常规生化试剂商店购买得到的。The present invention is further described below in conjunction with the examples, but these examples do not limit the scope of the present invention, and all simple improvements to the preparation method of the present invention under the conception of the present invention all fall within the protection scope of the present invention. The following examples do not specify the experimental method of specific conditions, usually according to the well-known means in the art. The test materials used in the following examples were purchased from conventional biochemical reagent stores unless otherwise specified.
以下实施例中制备中间体Int-1、Int-2如下所述：Preparation of intermediates Int-1 and Int-2 in the following examples are as follows:
Figure PCTCN2021127847-appb-000022
Figure PCTCN2021127847-appb-000022
合成路线如下：The synthetic route is as follows:
Figure PCTCN2021127847-appb-000023
Figure PCTCN2021127847-appb-000023
步骤1.1化合物1的制备Step 1.1 Preparation of Compound 1
Figure PCTCN2021127847-appb-000024
Figure PCTCN2021127847-appb-000024
把3-溴-1,2,4-三氮唑(化合物SM1,4.44g，30.0mmol)溶解于30ml DMF中并加入三乙烯二胺(6.73g，60.0mmol)，在室温下搅拌30分钟。然后加入(Z)-3-碘-丙烯酸异丙酯(化合物SM2,7.92g，33.0mmol)，并在室温下搅拌1h。将混合物倒入100mL冰水中，用乙酸乙酯(3x 50mL)提取水相。合并的有机相用50mL盐水洗涤两次，用无水硫酸钠干燥，过滤，减压浓缩，得到粗产品。将粗品制砂，用流动相(PE:EA＝5:1)洗脱，得到类白色固体1(6.25g，24.0mmol，产率80％)。3-Bromo-1,2,4-triazole (compound SM1, 4.44 g, 30.0 mmol) was dissolved in 30 ml of DMF, triethylenediamine (6.73 g, 60.0 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. Then (Z)-3-iodo-isopropyl acrylate (compound SM2, 7.92 g, 33.0 mmol) was added and stirred at room temperature for 1 h. The mixture was poured into 100 mL of ice water and the aqueous phase was extracted with ethyl acetate (3 x 50 mL). The combined organic phases were washed twice with 50 mL of brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product. The crude product was sanded and eluted with mobile phase (PE:EA=5:1) to give 1 (6.25 g, 24.0 mmol, 80% yield) as an off-white solid.
LCMS:t R＝1.222min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝259.8,261.7[M+H] +. LCMS: t R = 1.222 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 259.8, 261.7 [M+H] + .
步骤1.2化合物2的制备Step 1.2 Preparation of Compound 2
Figure PCTCN2021127847-appb-000025
Figure PCTCN2021127847-appb-000025
将化合物SM3(2.5g，10.4mmol)分散于氨水(50mL)溶液中，加入碘化钾(5.2g，31.2mmol)和碘单质(2.6g，10.4mmol)的水(25mL)溶液，室温下搅拌16小时。反应结束后，用浓盐酸将混合物调节至pH＝3并用EA(3×50mL)萃取。用饱和碳酸氢钠溶液(50mL)和食盐水(50mL)分别洗涤有机层。用无水硫酸钠干燥并浓缩，得到呈浅棕色油状的化合物4-溴-2-碘-6-(三氟甲基)苯酚(化合物2，3.0g，8.2mmol，产率78.9％)。Compound SM3 (2.5g, 10.4mmol) was dispersed in ammonia water (50mL) solution, potassium iodide (5.2g, 31.2mmol) and iodine element (2.6g, 10.4mmol) in water (25mL) solution were added, and stirred at room temperature for 16 hours . After the reaction was complete, the mixture was adjusted to pH=3 with concentrated hydrochloric acid and extracted with EA (3 x 50 mL). The organic layer was washed with saturated sodium bicarbonate solution (50 mL) and brine (50 mL), respectively. Drying over anhydrous sodium sulfate and concentration gave compound 4-bromo-2-iodo-6-(trifluoromethyl)phenol (compound 2, 3.0 g, 8.2 mmol, yield 78.9%) as light brown oil.
LCMS(ES,m/z):364.5[M-H]-,t R＝1.300min. LCMS (ES, m/z): 364.5 [MH]-, t R =1.300min.
步骤1.3化合物3的制备Step 1.3 Preparation of Compound 3
Figure PCTCN2021127847-appb-000026
Figure PCTCN2021127847-appb-000026
向4-溴-2-碘-6-(三氟甲基)苯酚(化合物2,3.3g，9.0mmol)和碳酸钾(3.7g，27.0mmol)的N,N-二甲基甲酰胺(40mL)溶液中加入3-溴-2-甲基丙-1-烯(化合物SM4,2.4g，18.0mmol)，室温下搅拌过夜。反应结束后，加水(400mL)淬灭并用乙酸乙酯(200mL×3)萃取。合并乙酸乙酯并用饱和食盐水(300mL)洗涤。用硫酸钠干燥并浓缩，得到棕色油状物5-溴-1-碘-2-((2-甲基烯丙基)氧基)-3-(三氟甲基)苯(化合物3,3.5g，8.3mmol，产率92.2％)。To 4-bromo-2-iodo-6-(trifluoromethyl)phenol (compound 2, 3.3 g, 9.0 mmol) and potassium carbonate (3.7 g, 27.0 mmol) in N,N-dimethylformamide (40 mL) ) solution was added 3-bromo-2-methylprop-1-ene (compound SM4, 2.4 g, 18.0 mmol), and the mixture was stirred at room temperature overnight. After the reaction was completed, it was quenched by adding water (400 mL) and extracted with ethyl acetate (200 mL×3). Ethyl acetate was combined and washed with saturated brine (300 mL). Dried over sodium sulfate and concentrated to give 5-bromo-1-iodo-2-((2-methylallyl)oxy)-3-(trifluoromethyl)benzene (compound 3, 3.5 g) as a brown oil , 8.3 mmol, 92.2% yield).
1H NMR(400MHz,CDCl 3)δ8.12(d,J＝2.3Hz,1H),7.72(d,J＝2.3Hz,1H),5.19(s,1H),5.04(s,1H),4.41(s,2H),1.91(s,3H). 1 H NMR (400 MHz, CDCl 3 ) δ 8.12 (d, J=2.3 Hz, 1H), 7.72 (d, J=2.3 Hz, 1H), 5.19 (s, 1H), 5.04 (s, 1H), 4.41 (s,2H),1.91(s,3H).
步骤1.4化合物4的制备Step 1.4 Preparation of Compound 4
Figure PCTCN2021127847-appb-000027
Figure PCTCN2021127847-appb-000027
向5-溴-1-碘-2-(丙-2-烯-1-基氧基)-3-(三氟甲基)苯(化合物3，15g，35.6mmol)的甲苯(200mL)溶液中分别加入氟化钾水溶液(6.2g，106.8mmol，10mL)和聚(甲基氢硅氧烷)(15.8 g，71.2mmol)，115℃搅拌10分钟，随后加入三丁基氯化锡(2.3g，7.1mmol)和偶氮二异丁腈(0.58g，3.6mmol)。所得混合物在115℃下搅拌1小时，冷却至室温然后用1M NaOH(200mL)淬灭并用乙酸乙酯(100mL×3)萃取。合并有机层，用饱和氯化铵(200mL)和食盐水(200mL)洗涤。用硫酸钠干燥并浓缩，得到粗品。制砂后，用石油醚/乙酸乙酯洗脱，得到无色油状物4(7g，23.7mmol，产率66.6％)。To a solution of 5-bromo-1-iodo-2-(prop-2-en-1-yloxy)-3-(trifluoromethyl)benzene (compound 3, 15 g, 35.6 mmol) in toluene (200 mL) Aqueous potassium fluoride solution (6.2 g, 106.8 mmol, 10 mL) and poly(methyl hydrogen siloxane) (15.8 g, 71.2 mmol) were added respectively, and the mixture was stirred at 115°C for 10 minutes, and then tributyltin chloride (2.3 g) was added. , 7.1 mmol) and azobisisobutyronitrile (0.58 g, 3.6 mmol). The resulting mixture was stirred at 115 °C for 1 hour, cooled to room temperature then quenched with 1 M NaOH (200 mL) and extracted with ethyl acetate (100 mL x 3). The organic layers were combined and washed with saturated ammonium chloride (200 mL) and brine (200 mL). Dried over sodium sulfate and concentrated to give crude product. After sand making, eluting with petroleum ether/ethyl acetate gave 4 (7 g, 23.7 mmol, 66.6% yield) as a colorless oil.
LCMS:t R＝1.539min,1.562min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),No MS. LCMS: t R = 1.539min, 1.562min in Shimadzu LC20, chromatography (HALO C18 2.7μm, 4.6mm×50mm), No MS.
步骤1.5化合物5的制备Step 1.5 Preparation of Compound 5
Figure PCTCN2021127847-appb-000028
Figure PCTCN2021127847-appb-000028
将化合物4(7g，23.7mmol)溶于二氧六环(150mL)溶液中，氮气保护下，分别加入联硼酸吡那醇酯(SM5，9.0g，35.5mmol)，醋酸钾(5.8g，59.3mmol)和1,1'-双(二苯基膦基)二茂铁-二氯化钯(2.6g，3.66mmol)，所得混合物在80℃下搅拌1小时，反应结束后加水(150mL)淬灭并用乙酸乙酯萃取(100mL x 3)，合并乙酸乙酯并用饱和食盐水(200mL)洗涤。用无水硫酸钠干燥并浓缩，得到粗品。通过硅胶柱色谱纯化，用石油醚/乙酸乙酯洗脱，得到粗品无色油状5(4.5g，13.2mmol，产率55.7％)。Compound 4 (7 g, 23.7 mmol) was dissolved in dioxane (150 mL) solution, and under nitrogen protection, pinacol biborate (SM5, 9.0 g, 35.5 mmol) and potassium acetate (5.8 g, 59.3 mmol) were added respectively. mmol) and 1,1'-bis(diphenylphosphino)ferrocene-palladium dichloride (2.6 g, 3.66 mmol), the resulting mixture was stirred at 80 °C for 1 hour, and water (150 mL) was added after the reaction to quench It was quenched and extracted with ethyl acetate (100 mL x 3), the ethyl acetate was combined and washed with saturated brine (200 mL). Dry over anhydrous sodium sulfate and concentrate to obtain crude product. Purification by silica gel column chromatography, eluting with petroleum ether/ethyl acetate, gave crude 5 as a colorless oil (4.5 g, 13.2 mmol, 55.7% yield).
LCMS:t R＝1.617min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),No MS. LCMS: t R = 1.617min in Shimadzu LC20, chromatography (HALO C18 2.7μm, 4.6mm×50mm), No MS.
步骤1.6化合物6的制备Step 1.6 Preparation of Compound 6
Figure PCTCN2021127847-appb-000029
Figure PCTCN2021127847-appb-000029
上一步得到的化合物5(12.7g，37.1mmol)溶于二氧六环/水＝10：1的溶液(200mL)中，分别加入异丙基(2Z)-3-(3-溴-1,2,4-三唑-1-基)2-烯酸丙酯(9.2g，35.2mmol)，碳酸钾(10.3g， 74.2mmol)及四(三苯基膦)钯(4.3g，3.71mmol)，氮气保护下80℃搅拌过夜。反应结束后，加水(200mL)淬灭，用乙酸乙酯(100mL×3)萃取。合并乙酸乙酯，并用饱和食盐水(200mL)洗涤。用硫酸钠干燥，浓缩得到粗产物。将其通过Combi-Flash纯化两次，用石油醚/乙酸乙酯(35∶1)洗脱，得到为黄色油状6(3.2g，8.1mmol，产率21.8％)。Compound 5 (12.7 g, 37.1 mmol) obtained in the previous step was dissolved in a solution (200 mL) of dioxane/water = 10:1, and isopropyl (2Z)-3-(3-bromo-1, 2,4-Triazol-1-yl)2-enoic acid propyl ester (9.2 g, 35.2 mmol), potassium carbonate (10.3 g, 74.2 mmol) and tetrakis(triphenylphosphine)palladium (4.3 g, 3.71 mmol) , and stirred at 80 °C overnight under nitrogen protection. After the reaction was completed, water (200 mL) was added to quench, and the mixture was extracted with ethyl acetate (100 mL×3). Ethyl acetate was combined and washed with saturated brine (200 mL). Dry over sodium sulfate and concentrate to give crude product. It was purified twice by Combi-Flash eluting with petroleum ether/ethyl acetate (35:1) to give 6 as a yellow oil (3.2 g, 8.1 mmol, 21.8% yield).
LCMS:t R＝1.515min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝396.2. LCMS: t R = 1.515 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm×50 mm), MS (ESI) m/z = 396.2.
1H NMR(300MHz,CDCl 3)δ9.70(s,1H),8.19(s,1H),8.04(s,1H),7.27–7.21(m,1H),5.68(d,J＝10.9Hz,1H),5.18–5.06(m,1H),4.45(s,2H),1.42(s,6H),1.32(d,J＝6.3Hz,6H). 1 H NMR (300MHz, CDCl 3 )δ9.70(s,1H), 8.19(s,1H), 8.04(s,1H), 7.27-7.21(m,1H), 5.68(d, J=10.9Hz, 1H), 5.18–5.06(m, 1H), 4.45(s, 2H), 1.42(s, 6H), 1.32(d, J=6.3Hz, 6H).
步骤1.7化合物7的制备Step 1.7 Preparation of Compound 7
Figure PCTCN2021127847-appb-000030
Figure PCTCN2021127847-appb-000030
将化合物6(4.9g，12.4mmol)溶解于二氯甲烷(80mL)中，在0℃下缓慢滴加溴素(3.95g，24.8mmol)。混合物在室温下搅拌过夜。反应完倒入冰水(80mL)中，用二氯甲烷(2x 80mL)提取。用饱和食盐水(3x 50mL)洗有机相，用无水硫酸钠干燥，减压浓缩，得到化合物7的黄色油状粗产品(6.8g，12.2mmol，粗产率98％),无需纯化，直接用于下一步。Compound 6 (4.9 g, 12.4 mmol) was dissolved in dichloromethane (80 mL), and bromine (3.95 g, 24.8 mmol) was slowly added dropwise at 0°C. The mixture was stirred at room temperature overnight. After the reaction was completed, it was poured into ice water (80 mL) and extracted with dichloromethane (2×80 mL). The organic phase was washed with saturated brine (3×50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a yellow oily crude product of compound 7 (6.8 g, 12.2 mmol, crude yield 98%), which was used directly without purification. in the next step.
LCMS:t R＝1.502min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝555.4[M+H] +. LCMS: t R = 1.502 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 555.4 [M+H] + .
步骤1.8化合物8的制备Step 1.8 Preparation of Compound 8
Figure PCTCN2021127847-appb-000031
Figure PCTCN2021127847-appb-000031
将2,3-二溴-3-(3,3-二甲基-7-(三氟甲基)-2,3-二氢苯并呋喃-5-基)-1H-1,2,4-三唑-1-基)丙酸 异丙酯(化合物7，6.8g，12.2mmol)溶解于四氢呋喃(80mL)中，0度下加入三乙胺(2.48g，24.5mmol)。将混合物在室温下搅拌15h。加入50ml冰水稀释混合物，用乙酸乙酯(3x 80mL)提取水层，然后用80mL盐水萃洗有机相两次，用无水硫酸钠干燥，过滤，并减压浓缩，得到淡黄色固体的粗品通过硅胶柱纯化，用石油醚/乙酸乙酯(8:1)洗脱，得到白色固体8(3.0g，6.3mmol，产率51.9％)。2,3-Dibromo-3-(3,3-dimethyl-7-(trifluoromethyl)-2,3-dihydrobenzofuran-5-yl)-1H-1,2,4 Isopropyl triazol-1-yl)propionate (compound 7, 6.8 g, 12.2 mmol) was dissolved in tetrahydrofuran (80 mL), and triethylamine (2.48 g, 24.5 mmol) was added at 0 degrees. The mixture was stirred at room temperature for 15 h. 50 ml of ice water was added to dilute the mixture, the aqueous layer was extracted with ethyl acetate (3 x 80 mL), then the organic phase was washed twice with 80 mL of brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product as a pale yellow solid Purification by silica gel column eluting with petroleum ether/ethyl acetate (8:1) gave 8 as a white solid (3.0 g, 6.3 mmol, 51.9% yield).
LCMS:t R＝1.225min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝473.6，475.6[M+H] +. LCMS: t R = 1.225 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 473.6, 475.6 [M+H] + .
1H NMR(400MHz,DMSO-d6)δ9.38(s,1H),8.86(s,1H),8.13(s,1H),8.07(s,1H),5.12～5.06(m,1H),4.51(s,2H),1.39(s,6H),1.33(d,J＝6.0Hz,6H). 1 H NMR(400MHz,DMSO-d6)δ9.38(s,1H),8.86(s,1H),8.13(s,1H),8.07(s,1H),5.12～5.06(m,1H),4.51 (s, 2H), 1.39(s, 6H), 1.33(d, J=6.0Hz, 6H).
步骤1.9化合物Int-1的制备Step 1.9 Preparation of compound Int-1
Figure PCTCN2021127847-appb-000032
Figure PCTCN2021127847-appb-000032
将化合物8(2000mg，4.2mmol)溶解于THF(30ml)中。冷却至0度缓慢滴加一水氢氧化锂(224mg，5.5mmol)和15mL水配成的溶液。反应混合液在0℃搅拌1h。反应液减压浓缩，剩余物用15mL水稀释。用甲基叔丁基醚(2x 50mL)萃取，然后用浓盐酸将水相调节到pH＝2，用乙酸乙酯3x 50ml提取水层。用无水硫酸钠干燥，过滤，并减压浓缩，得到白色固体Int-1(1230mg，2.85mmol，产率67.9％)。Compound 8 (2000 mg, 4.2 mmol) was dissolved in THF (30 ml). Cool to 0 degrees and slowly add a solution of lithium hydroxide monohydrate (224 mg, 5.5 mmol) and 15 mL of water dropwise. The reaction mixture was stirred at 0 °C for 1 h. The reaction solution was concentrated under reduced pressure, and the residue was diluted with 15 mL of water. Extracted with methyl tert-butyl ether (2 x 50 mL), then the aqueous phase was adjusted to pH=2 with concentrated hydrochloric acid, and the aqueous layer was extracted with ethyl acetate 3 x 50 ml. Dry over anhydrous sodium sulfate, filter, and concentrate under reduced pressure to give Int-1 as a white solid (1230 mg, 2.85 mmol, 67.9% yield).
LCMS:t R＝1.330min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝431.6，433.6[M+H] +. LCMS: t R = 1.330 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 431.6, 433.6 [M+H] + .
1H NMR(400MHz,DMSO-d 6)δ13.98(brs,1H),9.35(s,1H),8.85(s,1H),8.12(s,1H),8.07(s,1H),4.50(s,2H),1.39(s,6H). 1 H NMR (400MHz, DMSO-d 6 )δ13.98(brs,1H), 9.35(s,1H), 8.85(s,1H), 8.12(s,1H), 8.07(s,1H), 4.50( s,2H),1.39(s,6H).
步骤1.10化合物Int-2的制备Step 1.10 Preparation of compound Int-2
Figure PCTCN2021127847-appb-000033
Figure PCTCN2021127847-appb-000033
将化合物Int-1(1g，2.31mmol)溶解在30mL THF中，0度下添加N-甲基吗啡啉(350mg，3.47mmol)和氯甲酸异丙酯(196mg，1.6mmol)。反应混合液在0度搅拌1小时后加入氨的甲醇溶液(3mL，7M)，继续搅拌30分钟。所得到的混合物在真空下浓缩，剩余物用10mL冰水稀释，水相用乙酸乙酯3x50ml萃取并用盐水洗有机相，并用无水硫酸钠干燥并在真空下浓缩，得到类白色固体(880mg，0.68mmol，产率88.4％)。Compound Int-1 (1 g, 2.31 mmol) was dissolved in 30 mL of THF, and N-methylmorpholine (350 mg, 3.47 mmol) and isopropyl chloroformate (196 mg, 1.6 mmol) were added at 0 degrees. The reaction mixture was stirred at 0 degrees for 1 hour, then methanol solution of ammonia (3 mL, 7M) was added, and stirring was continued for 30 minutes. The resulting mixture was concentrated under vacuum, the residue was diluted with 10 mL of ice water, the aqueous phase was extracted with ethyl acetate 3×50 ml and the organic phase was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum to give an off-white solid (880 mg, 0.68 mmol, 88.4% yield).
LCMS:t R＝1.276min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝430.6，432.6[M+H] +. LCMS: t R = 1.276 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 430.6, 432.6 [M+H] + .
实施例1：化合物P-1的制备Example 1: Preparation of Compound P-1
Figure PCTCN2021127847-appb-000034
Figure PCTCN2021127847-appb-000034
将化合物Int-2(862mg，2.0mmol)溶于二氧六环/水的混合溶剂33mL(10：1)中，分别加入5-硼酸嘧啶(371.7mg，3.0mmol)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(245mg，0.3mmol)，乙酸钾(588mg，6.0mmol)，氮气保护下85℃搅拌10小时。反应结束后，将反应液浓缩。以纯EA为展开剂，采用prep-TLC(洗脱剂：乙酸乙酯)和prep-HPLC进行分离纯化，得到白色固体目标产物(350mg,0.81mmol,产率40.7％)。Compound Int-2 (862 mg, 2.0 mmol) was dissolved in 33 mL (10:1) of a mixed solvent of dioxane/water, and 5-boronic acid pyrimidine (371.7 mg, 3.0 mmol), [1,1'- Bis(diphenylphosphino)ferrocene]palladium dichloride (245 mg, 0.3 mmol), potassium acetate (588 mg, 6.0 mmol), stirred at 85°C for 10 hours under nitrogen protection. After completion of the reaction, the reaction solution was concentrated. Using pure EA as a developing solvent, separation and purification were carried out by prep-TLC (eluent: ethyl acetate) and prep-HPLC to obtain the target product (350 mg, 0.81 mmol, yield 40.7%) as a white solid.
LCMS:t R＝1.170min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝430.7[M+H] +. LCMS: t R = 1.170 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 430.7 [M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.21(s,1H),9.01(s,1H),8.71(s,2H),8.38(s,1H),7.59～7.58(m, 3H),7.35(s,1H),4.47(s,2H),1.33(s,6H). 1 H NMR (400MHz, DMSO-d 6 )δ9.21(s, 1H), 9.01(s, 1H), 8.71(s, 2H), 8.38(s, 1H), 7.59～7.58(m, 3H), 7.35(s, 1H), 4.47(s, 2H), 1.33(s, 6H).
实施例2：化合物P-2的制备Example 2: Preparation of Compound P-2
Figure PCTCN2021127847-appb-000035
Figure PCTCN2021127847-appb-000035
将化合物Int-2(400mg,0.928mmol)溶于二氧六环/水的混合溶剂33mL(10：1)中，分别加入4-吡啶硼酸(228.04mg,1.855mmol)，乙酸钾(273.10mg,2.783mmol)及[1,1'-双(二苯基膦)二茂铁]二氯化钯(67.87mg,0.0927mmol)。反应混合物在氮气保护下85℃搅拌10h，过滤，减压浓缩。采用prep-TLC(洗脱剂：乙酸乙酯)和prep-HPLC进行分离纯化，得到(E)-3-(3-(3,3-二甲基-7-(三氟甲基)-2,3-二氢苯并呋喃-5-基)-1h-1,2,4-三唑-1-基)-2-(吡啶-4-基)丙烯酰胺(37.5mg，纯度99.23％，产率9.35％)，白色固体。Compound Int-2 (400 mg, 0.928 mmol) was dissolved in 33 mL (10:1) of a mixed solvent of dioxane/water, 4-pyridineboronic acid (228.04 mg, 1.855 mmol), potassium acetate (273.10 mg, 1.855 mmol) were added respectively. 2.783 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloride palladium (67.87 mg, 0.0927 mmol). The reaction mixture was stirred at 85°C for 10 h under nitrogen protection, filtered and concentrated under reduced pressure. Using prep-TLC (eluent: ethyl acetate) and prep-HPLC for separation and purification to obtain (E)-3-(3-(3,3-dimethyl-7-(trifluoromethyl)-2) ,3-Dihydrobenzofuran-5-yl)-1h-1,2,4-triazol-1-yl)-2-(pyridin-4-yl)acrylamide (37.5 mg, 99.23% pure, produced rate 9.35%), white solid.
LCMS:t R＝1.075min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝429.7[M+H] +. LCMS: t R = 1.075 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 429.7 [M+H] + .
HPLC:t R＝4.701min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =4.701min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(301MHz,DMSO-d 6)δ8.79(s,1H),8.61(dd,J＝4.5,1.5Hz,2H),8.18(s,1H),7.62(s,1H),7.56(s,1H),7.50(s,1H),7.27(dd,J＝4.5,1.5Hz,2H),7.09(s,1H),4.44(s,2H),1.31(s,6H). 1H NMR (301MHz, DMSO-d 6 ) δ 8.79(s, 1H), 8.61(dd, J=4.5, 1.5Hz, 2H), 8.18(s, 1H), 7.62(s, 1H), 7.56(s ,1H),7.50(s,1H),7.27(dd,J=4.5,1.5Hz,2H),7.09(s,1H),4.44(s,2H),1.31(s,6H).
实施例3：化合物P-3的制备Example 3: Preparation of Compound P-3
Figure PCTCN2021127847-appb-000036
Figure PCTCN2021127847-appb-000036
将化合物Int-2(50mg,0.116mmol)溶于二氧六环/水的混合溶剂11mL(10：1)中，分别加入化合物SM8(34.63mg,0.174mmol)，醋酸钾(22.77mg,0.232mmol)及1,1-双(二苯基磷)二茂铁氯化钯(14.2mg,0.0174mmol).。反应液在氮气保护下80℃搅拌过夜，反应结束后，硅藻土过滤,减压浓 缩，采用prep-TLC(洗脱剂：乙酸乙酯)和prep-HPLC进行分离纯化，得到灰色固体P-3(13mg,98.192％纯度,产率21.81％).Compound Int-2 (50 mg, 0.116 mmol) was dissolved in 11 mL (10:1) of a mixed solvent of dioxane/water, and compound SM8 (34.63 mg, 0.174 mmol), potassium acetate (22.77 mg, 0.232 mmol) were added respectively. ) and 1,1-bis(diphenylphosphonium)ferrocene palladium chloride (14.2 mg, 0.0174 mmol). The reaction solution was stirred overnight at 80°C under nitrogen protection. After the reaction was completed, celite was filtered, concentrated under reduced pressure, and separated and purified by prep-TLC (eluent: ethyl acetate) and prep-HPLC to obtain a gray solid P- 3 (13 mg, 98.192% purity, 21.81% yield).
LCMS:t R＝1.097min in 10-80AB_7min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*30mm),MS(ESI)m/z＝505.6[M+H] +. LCMS: t R =1.097min in 10-80AB_7min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*30mm),MS(ESI)m/z=505.6[M+H] + .
HPLC:t R＝3.216min in 10-80AB_15min.met,chromatography(YMC-TriartC18 ExRS 150*4.6mml.D S-5um). HPLC: t R =3.216min in 10-80AB_15min.met,chromatography (YMC-TriartC18 ExRS 150*4.6mml.D S-5um).
1H NMR(400MHz,DMSO-d 6):δ9.02(d,J＝1.7Hz,1H),8.63(s,1H),8.62–8.59(m,1H),8.25–8.17(m,1H),8.15(s,1H),7.88(d,J＝8.4Hz,2H),7.72(d,J＝1.0Hz,1H),7.65(s,1H),7.61–7.48(m,2H),7.40(d,J＝8.4Hz,2H),7.07(s,1H),4.40(s,2H),1.16(s,6H). 1 H NMR (400MHz, DMSO-d 6 ): δ 9.02 (d, J=1.7 Hz, 1H), 8.63 (s, 1H), 8.62-8.59 (m, 1H), 8.25-8.17 (m, 1H) ,8.15(s,1H),7.88(d,J=8.4Hz,2H),7.72(d,J=1.0Hz,1H),7.65(s,1H),7.61–7.48(m,2H),7.40( d, J=8.4Hz, 2H), 7.07(s, 1H), 4.40(s, 2H), 1.16(s, 6H).
实施例4：化合物P-4的制备Example 4: Preparation of Compound P-4
Figure PCTCN2021127847-appb-000037
Figure PCTCN2021127847-appb-000037
化合物Int-2(50mg，0.12mmol)溶于二氧六环/水的混合溶剂11mL(10：1)中，分别加入喹啉-3-基硼烷二醇(30mg，0.17mmmol)，乙酸钾(23mg，0.23mmol)及[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(14mg，0.017mmol)，反应液在氮气下于80℃中搅拌过夜。反应结束后，将混合物用水(10mL)稀释，并用乙酸乙酯(3×10mL)萃取。用饱和盐水(20mL)洗涤有机相，并用硫酸钠干燥。浓缩得到残留物。通过用展开剂(石油醚/乙酸乙酯＝1：3)的硅胶预制备板纯化得到粗品，并通过高效制备液相色谱纯化(流动相：
Figure PCTCN2021127847-appb-000038
)，得到白色固体产物P-4(20mg，0.042mmol，产率36.0％)。 Compound Int-2 (50 mg, 0.12 mmol) was dissolved in 11 mL (10:1) of a mixed solvent of dioxane/water, and quinolin-3-ylboranediol (30 mg, 0.17 mmol) and potassium acetate were added respectively. (23 mg, 0.23 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (14 mg, 0.017 mmol), the reaction solution was at 80°C under nitrogen Stir overnight. After the reaction was complete, the mixture was diluted with water (10 mL) and extracted with ethyl acetate (3 x 10 mL). The organic phase was washed with saturated brine (20 mL) and dried over sodium sulfate. Concentration gave a residue. The crude product was purified by preparative silica gel plate purification with developing solvent (petroleum ether/ethyl acetate = 1:3) and purified by high performance preparative liquid chromatography (mobile phase:
Figure PCTCN2021127847-appb-000038
) to give the product P-4 as a white solid (20 mg, 0.042 mmol, 36.0% yield).
LCMS:t R＝1.168min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝479.7[M+H] +. LCMS: t R = 1.168 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 479.7 [M+H] + .
HPLC:t R＝5.307min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =5.307min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(400MHz,DMSO-d 6)δ8.97(s,1H),8.73(d,J＝2.2Hz,1H),8.38(s,1H),8.28(d,J＝2.0Hz,1H),8.08(d,J＝8.4Hz,1H),8.02(dd,J＝8.3,0.7Hz,1H),7.85-7.78(m,1H),7.66–7.61(m,1H), 7.58(s,1H),7.53(d,J＝1.1Hz,1H),7.24(s,1H),7.19(d,J＝1.1Hz,1H),4.37(s,2H),1.09(s,6H). 1 H NMR (400MHz, DMSO-d 6 ) δ 8.97(s, 1H), 8.73(d, J=2.2Hz, 1H), 8.38(s, 1H), 8.28(d, J=2.0Hz, 1H) ,8.08(d,J＝8.4Hz,1H),8.02(dd,J＝8.3,0.7Hz,1H),7.85-7.78(m,1H),7.66-7.61(m,1H), 7.58(s,1H) ), 7.53(d, J=1.1Hz, 1H), 7.24(s, 1H), 7.19(d, J=1.1Hz, 1H), 4.37(s, 2H), 1.09(s, 6H).
实施例5：化合物P-5的制备Example 5: Preparation of Compound P-5
Figure PCTCN2021127847-appb-000039
Figure PCTCN2021127847-appb-000039
将化合物Int-2(50mg，0.12mmol)溶于二氧六环/水的混合溶剂11mL(10：1)中，分别加入(2-氟-[1,1'-联苯]-4-基)硼酸(SM10)(37.6mg，0.17mmmol)，乙酸钾(28.46mg，0.29mmol)及[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(14mg，0.017mmol)，反应液在氮气保护下于80℃下搅拌16小时。反应结束后，加水(5mL)稀释，用EA(3×10mL)萃取混合物。用饱和盐水(30mL)洗涤有机相，并用硫酸钠干燥。浓缩得到粗品。通过用展开剂(石油醚/乙酸乙酯＝1:3)的硅胶预制备板纯化得到粗品，并通过高效制备液相色谱(流动相：乙腈/水：
Figure PCTCN2021127847-appb-000040
)纯化，得到白色固体目标产物P-5(17.8mg，0.034mmol，产率29.4％)。 Compound Int-2 (50 mg, 0.12 mmol) was dissolved in 11 mL (10:1) of a mixed solvent of dioxane/water, and (2-fluoro-[1,1'-biphenyl]-4-yl) was added respectively. ) boric acid (SM10) (37.6 mg, 0.17 mmol), potassium acetate (28.46 mg, 0.29 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (14 mg, 0.017 mmol), and the reaction was stirred at 80° C. for 16 hours under nitrogen protection. After the reaction was completed, water (5 mL) was added to dilute, and the mixture was extracted with EA (3 x 10 mL). The organic phase was washed with saturated brine (30 mL) and dried over sodium sulfate. Concentration gave crude product. The crude product was obtained by purification on a silica gel preparative plate with a developing solvent (petroleum ether/ethyl acetate=1:3) and subjected to high performance preparative liquid chromatography (mobile phase: acetonitrile/water:
Figure PCTCN2021127847-appb-000040
) was purified to obtain the desired product P-5 (17.8 mg, 0.034 mmol, 29.4% yield) as a white solid.
LCMS:t R＝1.372min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝522.7[M+H] +. LCMS: t R = 1.372 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 522.7 [M+H] + .
HPLC:t R＝5.307min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =5.307min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(400MHz,DMSO-d 6)δ8.74(s,1H),8.18(s,1H),7.75(d,J＝1.0Hz,1H),7.69(d,J＝1.0Hz,1H),7.66–7.54(m,4H),7.51(t,J＝7.5Hz,2H),7.47–7.39(m,1H),7.27(dd,J＝11.8,1.5Hz,1H),7.17(dd,J＝7.9,1.6Hz,1H),7.12(s,1H),4.41(s,2H),1.19(s,6H). 1 H NMR (400MHz, DMSO-d 6 ) δ 8.74 (s, 1H), 8.18 (s, 1H), 7.75 (d, J=1.0 Hz, 1H), 7.69 (d, J=1.0 Hz, 1H) , 7.66–7.54 (m, 4H), 7.51 (t, J=7.5Hz, 2H), 7.47–7.39 (m, 1H), 7.27 (dd, J=11.8, 1.5Hz, 1H), 7.17 (dd, J =7.9,1.6Hz,1H),7.12(s,1H),4.41(s,2H),1.19(s,6H).
实施例6：化合物P-6的制备Example 6: Preparation of Compound P-6
Figure PCTCN2021127847-appb-000041
Figure PCTCN2021127847-appb-000041
(Z)-2-溴-3-(3-(3,3-二甲基-7-(三氟甲基)-2,3-二氢苯并呋喃-5-基)-1h-1,2,4-三唑-1-基)丙烯酰胺Int-2(70mg,0.162mmol)溶于二氧六环/水的混合溶剂11mL(10：1)中，分别加入苯硼酸SM11(39.58mg,0.325mmol)，乙酸钾(47.78mg,0.487mmol)及[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(13.25mg,0.0162mmol)。反应混合物在氮气保护下80℃搅拌10h，然后过滤，减压浓缩。采用prep-TLC(洗脱剂：石油醚/乙酸乙酯＝1/2)和prep-HPLC进行分离纯化，得到白色固体(E)-3-(3-(3,3-二甲基-7-(三氟甲基)-2,3-二氢苯并呋喃-5-基)-1H-1,2,4-三唑-1-基-2-苯丙烯酰胺(21.7mg，纯度99.25％，产率31.0％)。(Z)-2-Bromo-3-(3-(3,3-dimethyl-7-(trifluoromethyl)-2,3-dihydrobenzofuran-5-yl)-1h-1, 2,4-Triazol-1-yl)acrylamide Int-2 (70mg, 0.162mmol) was dissolved in 11mL (10:1) of a mixed solvent of dioxane/water, and phenylboronic acid SM11 (39.58mg, 0.325 mmol), potassium acetate (47.78 mg, 0.487 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (13.25 mg, 0.0162 mmol). The reaction mixture was stirred at 80°C for 10 h under nitrogen protection, then filtered and concentrated under reduced pressure. Using prep-TLC (eluent: petroleum ether/ethyl acetate=1/2) and prep-HPLC for separation and purification, white solid (E)-3-(3-(3,3-dimethyl-7) was obtained -(Trifluoromethyl)-2,3-dihydrobenzofuran-5-yl)-1H-1,2,4-triazol-1-yl-2-phenylacrylamide (21.7 mg, purity 99.25%) , yield 31.0%).
LCMS:t R＝1.256min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝428.7[M+H] +. LCMS: t R = 1.256 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 428.7 [M+H] + .
HPLC:t R＝4.875min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =4.875min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(300MHz,DMSO-d 6)δ8.34(s,1H),8.06(s,1H),7.70(d,J＝2.2Hz,2H),7.53(s,1H),7.46–7.41(m,3H),7.23(dd,J＝6.9,2.7Hz,2H),6.95(s,1H),4.44(s,2H),1.31(s,6H). 1 H NMR (300MHz, DMSO-d 6 )δ8.34(s,1H),8.06(s,1H),7.70(d,J=2.2Hz,2H),7.53(s,1H),7.46-7.41( m, 3H), 7.23(dd, J=6.9, 2.7Hz, 2H), 6.95(s, 1H), 4.44(s, 2H), 1.31(s, 6H).
实施例7：化合物P-7的制备Example 7: Preparation of compound P-7
Figure PCTCN2021127847-appb-000042
Figure PCTCN2021127847-appb-000042
(Z)-2-溴-3-(3-(3,3-二甲基-7-(三氟甲基)-2,3-二氢苯并呋喃-5-基)-1H-1,2,4-三唑-1-基)丙烯酰胺Int-2(60mg，0.14mmol)溶于二氧六环/水的混合溶剂11mL(10：1)中，分别加入(5-氟吡啶-3-基)硼烷二醇SM12(40mg,0.28mmol)，乙酸钾(29mg，0.21mmol)及[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(17mg，0.02mmol)，反应液在80℃下搅拌16小时。反应结束后加水(5mL)稀释，并将混合物用乙酸乙酯(3×10mL)萃取。用饱和食盐水(30mL)洗涤有机相，并用硫酸钠干燥。浓缩得到粗品。通过用展开剂(二氯甲烷/甲醇＝10:1)的硅胶预制备板纯化，然后通过高效制备液相色谱(流动相：乙腈/水：
Figure PCTCN2021127847-appb-000043
)纯化，得到白色固体目标产物P-7(15mg，0.033mmol，产率24.1％)。 (Z)-2-Bromo-3-(3-(3,3-dimethyl-7-(trifluoromethyl)-2,3-dihydrobenzofuran-5-yl)-1H-1, 2,4-Triazol-1-yl)acrylamide Int-2 (60 mg, 0.14 mmol) was dissolved in 11 mL (10:1) of a mixed solvent of dioxane/water, and (5-fluoropyridine-3 -yl)boranediol SM12 (40mg, 0.28mmol), potassium acetate (29mg, 0.21mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex Compound (17 mg, 0.02 mmol), the reaction solution was stirred at 80°C for 16 hours. After the reaction was completed, water (5 mL) was added to dilute, and the mixture was extracted with ethyl acetate (3 x 10 mL). The organic phase was washed with saturated brine (30 mL) and dried over sodium sulfate. Concentration gave crude product. Purification by preparative silica gel plate with developing solvent (dichloromethane/methanol=10:1) followed by high performance preparative liquid chromatography (mobile phase: acetonitrile/water:
Figure PCTCN2021127847-appb-000043
) was purified to obtain the desired product P-7 as a white solid (15 mg, 0.033 mmol, 24.1% yield).
LCMS:t R＝1.272min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm), MS(ESI)m/z＝448.2[M+H] +. LCMS: t R =1.272min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm), MS(ESI)m/z=448.2[M+H] + .
HPLC:t R＝6.262min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =6.262min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(400MHz,DMSO-d 6)δ8.94(s,1H),8.60(d,J＝2.7Hz,1H),8.32(s,1H),8.30(t,J＝1.6Hz,1H),7.78–7.70(m,1H),7.60(s,3H),7.18(s,1H),4.46(s,2H),1.32(s,6H). 1 H NMR (400MHz, DMSO-d 6 ) δ 8.94(s, 1H), 8.60(d, J=2.7Hz, 1H), 8.32(s, 1H), 8.30(t, J=1.6Hz, 1H) ,7.78–7.70(m,1H),7.60(s,3H),7.18(s,1H),4.46(s,2H),1.32(s,6H).
实施例8：化合物P-8的制备Example 8: Preparation of Compound P-8
Figure PCTCN2021127847-appb-000044
Figure PCTCN2021127847-appb-000044
采用实施例1的方法，得到目标产物P-8。Using the method of Example 1, the target product P-8 was obtained.
LCMS:t R＝1.137min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝429.7[M+H] +. LCMS: t R = 1.137 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 429.7 [M+H] + .
HPLC:t R＝5.546min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =5.546min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(400MHz,MeOD)δ8.66(s,1H),8.64(d,J＝4.0Hz，1H),8.51(s,1H),8.30(s，1H),7.84～7.81(m,1H),7.69(s,1H),7.63(s,1H),7.60～7.56(m,1H),4.42(s,2H),1.37(s,6H). 1 H NMR(400MHz,MeOD)δ8.66(s,1H),8.64(d,J=4.0Hz,1H),8.51(s,1H),8.30(s,1H),7.84～7.81(m,1H) ),7.69(s,1H),7.63(s,1H),7.60～7.56(m,1H),4.42(s,2H),1.37(s,6H).
实施例9：化合物P-9的制备Example 9: Preparation of Compound P-9
Figure PCTCN2021127847-appb-000045
Figure PCTCN2021127847-appb-000045
采用实施例1的方法，得到目标产物P-9。MS(ESI)m/z＝444.3[M+H] + Using the method of Example 1, the target product P-9 was obtained. MS(ESI)m/z=444.3[M+H] +
实施例10：化合物P-10的制备Example 10: Preparation of Compound P-10
Figure PCTCN2021127847-appb-000046
Figure PCTCN2021127847-appb-000046
将4-(5-溴嘧啶-2-基)吗啉(SM13,250mg，1.02mmol)溶于二氧六环(10mL)中，分别加入联硼酸频哪醇酯(442mg，1.74mmol)，乙酸钾(251mg，2.56mmol)及[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(84mg，0.10mmol)，反应液在氮气保护下于85℃搅拌过夜。反应结束后，加入冰水(10mL)，并用乙酸乙酯(3×20mL)萃取。用饱和盐水(30mL)洗涤有机相，并用硫酸钠干燥，浓缩得到残留物。将其通过Combi-Flash纯化，用石油醚/乙酸乙酯(3∶1)洗脱，得到白色固体9(200mg，0.67mmol，产率65.7％)；将化合物Int-2(80mg，0.18mmol)溶于二氧六环/水的混合溶剂11mL(10：1)中，分别加入4-[5-(4,4,5,5-四甲基-1,3,2-二氧杂硼硼烷-2-基)嘧啶-2-基]吗啉(化合物9，81mg，0.28mmol)，乙酸钾(45mg，0.46mmol)及[1,1'-双(二苯基膦基)二茂铁]二氯化钯(20mg，0.028mmol)，反应液在氮气保护下于80℃搅拌过夜。反应结束后，加冰水(10mL)稀释，并将混合物用乙酸乙酯(3×10mL)萃取。用饱和盐水(20mL)洗涤有机相，并用硫酸钠干燥。浓缩得到残留物。用展开剂(石油醚/乙酸乙酯)的硅胶预制备板纯化得到粗品，并通过高效制备液相色谱纯化(流动相：乙腈/水＝35/65～70/30)，得到白色固体化合物P-10(12mg，0.023mmol，产率12.3％)。4-(5-Bromopyrimidin-2-yl)morpholine (SM13, 250 mg, 1.02 mmol) was dissolved in dioxane (10 mL), pinacol biboronate (442 mg, 1.74 mmol) and acetic acid were added respectively. Potassium (251mg, 2.56mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (84mg, 0.10mmol), the reaction solution was heated at 85 under nitrogen protection Stir overnight at °C. After the reaction was completed, ice water (10 mL) was added and extracted with ethyl acetate (3×20 mL). The organic phase was washed with saturated brine (30 mL), dried over sodium sulfate, and concentrated to give a residue. It was purified by Combi-Flash eluting with petroleum ether/ethyl acetate (3:1) to give 9 (200 mg, 0.67 mmol, 65.7% yield) as a white solid; compound Int-2 (80 mg, 0.18 mmol) Dissolve in 11 mL (10:1) of a mixed solvent of dioxane/water, and add 4-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborole respectively) Alk-2-yl)pyrimidin-2-yl]morpholine (compound 9, 81 mg, 0.28 mmol), potassium acetate (45 mg, 0.46 mmol) and [1,1'-bis(diphenylphosphino)ferrocene ] Palladium dichloride (20 mg, 0.028 mmol), the reaction solution was stirred at 80° C. overnight under nitrogen protection. After the reaction was completed, ice water (10 mL) was added to dilute, and the mixture was extracted with ethyl acetate (3 x 10 mL). The organic phase was washed with saturated brine (20 mL) and dried over sodium sulfate. Concentration gave a residue. The crude product was purified by silica gel preparative plate with developing solvent (petroleum ether/ethyl acetate), and purified by high performance liquid chromatography (mobile phase: acetonitrile/water=35/65～70/30) to obtain white solid compound P -10 (12 mg, 0.023 mmol, 12.3% yield).
LCMS:t R＝1.289min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝516.2[M+H] +. LCMS: t R = 1.289 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 516.2 [M+H] + .
HPLC:t R＝5.997min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =5.997min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(400MHz,DMSO-d 6)δ8.95(s,1H),8.26(s,2H),8.20(s,1H),7.75(d,J＝12.6Hz,2H),7.40(d,J＝67.3Hz,2H),4.48(s,2H),3.86–3.57(m,8H),1.34(s,6H). 1 H NMR (400MHz, DMSO-d 6 ) δ 8.95(s, 1H), 8.26(s, 2H), 8.20(s, 1H), 7.75(d, J=12.6Hz, 2H), 7.40(d, J=67.3Hz, 2H), 4.48(s, 2H), 3.86–3.57(m, 8H), 1.34(s, 6H).
实施例11：化合物P-11的制备Example 11: Preparation of compound P-11
Figure PCTCN2021127847-appb-000047
Figure PCTCN2021127847-appb-000047
5-溴-N-甲基嘧啶-2-胺(化合物SM14,500mg,2.66mmol)溶于DMF(20mL)中，0℃下加入氢化钠(78.5mg,3.27mmol)，反应0.5h，加入乙基磺酰氯(化合物SM15,410.3mg,3.19mmol)，室温搅拌16h。倒入冰水(20mL)中，乙酸乙酯(20mL×3)萃取，有机相盐水(30mL×2)洗涤，无水硫酸钠干燥，过滤，浓缩。采用prep-HPLC进行分离提纯，得到化合物10(245mg，产率32.89％)，白色固体LCMS:t R＝1.249min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝279.9,281.9[M+H] +；将化合物10(245mg,0.875mmol)、联硼酸频那醇酯(333.14mg,1.31mmol)和乙酸钾(257.5mg,2.62mmol)溶于二氧六环(50mL)中，加入[1,1'-双(二苯基膦)二茂铁]二氯化钯(64mg,0.087mmol)。在氮气保护下100℃搅拌16h，然后过滤，减压浓缩。采用flash色谱法(洗脱剂：0％～40％乙酸乙酯/石油醚)进行分离纯化。将含有所需要的化合物的馏分蒸发并干燥，得到(2-(N-甲基乙基磺酰胺)嘧啶-5-基)硼酸(化合物11，200mg，产率93.31％)，白色固体；将化合物Int-2(100mg,0.232mmol)、(2-(N-甲基乙基磺酰胺)嘧啶-5-基)硼酸(化合物11,113.66mg,0.464mmol)和乙酸钾(68.28mg,0.696mmol)溶于二氧六环/水(11mL,v/v＝10:1)中，加入[1,1'-双(二苯基膦)二茂铁]二氯化钯(16.97mg,0.023mmol)。反应混合物在氮气保护下85℃搅拌10h，然后过滤，减压浓缩。采用prep-TLC(洗脱剂：乙酸乙酯)和prep-HPLC进行分离纯化，得到化合物P-11(22.4mg，纯度97.67％，产率17.12％)，白色固体。 5-Bromo-N-methylpyrimidin-2-amine (compound SM14, 500 mg, 2.66 mmol) was dissolved in DMF (20 mL), sodium hydride (78.5 mg, 3.27 mmol) was added at 0 °C, the reaction was carried out for 0.5 h, and ethyl acetate was added. sulfonyl chloride (compound SM15, 410.3 mg, 3.19 mmol), stirred at room temperature for 16 h. Pour into ice water (20 mL), extract with ethyl acetate (20 mL×3), wash the organic phase with brine (30 mL×2), dry over anhydrous sodium sulfate, filter and concentrate. Separation and purification by prep-HPLC gave compound 10 (245 mg, yield 32.89%), white solid LCMS: t R =1.249 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm×50 mm), MS (ESI) m/z = 279.9, 281.9 [M+H] + ; Compound 10 (245 mg, 0.875 mmol), pinacol diboronate (333.14 mg, 1.31 mmol) and potassium acetate (257.5 mg, 2.62 mmol) were dissolved in two To oxane (50 mL) was added [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (64 mg, 0.087 mmol). Stir at 100°C for 16 h under nitrogen protection, then filter and concentrate under reduced pressure. It was separated and purified by flash chromatography (eluent: 0%-40% ethyl acetate/petroleum ether). The fractions containing the desired compound were evaporated and dried to give (2-(N-methylethylsulfonamide)pyrimidin-5-yl)boronic acid (compound 11, 200 mg, 93.31% yield) as a white solid; the compound Int-2 (100 mg, 0.232 mmol), (2-(N-methylethylsulfonamide)pyrimidin-5-yl)boronic acid (compound 11, 113.66 mg, 0.464 mmol) and potassium acetate (68.28 mg, 0.696 mmol) were dissolved in [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (16.97 mg, 0.023 mmol) was added to dioxane/water (11 mL, v/v=10:1). The reaction mixture was stirred at 85°C for 10 h under nitrogen protection, then filtered and concentrated under reduced pressure. Separation and purification were carried out by prep-TLC (eluent: ethyl acetate) and prep-HPLC to obtain compound P-11 (22.4 mg, purity 97.67%, yield 17.12%) as a white solid.
LCMS:t R＝1.325min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝552.2[M+H] +. LCMS: t R = 1.325 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 552.2 [M+H] + .
HPLC:t R＝6.670min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =6.670min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(301MHz,DMSO-d6)δ9.01(s,1H),8.57(d,J＝1.6Hz,2H),8.33(s,1H),7.67(d,J＝6.8Hz,2H),7.57(s,1H),7.38(s,1H),4.45(s,2H),3.83–3.75(m,2H),3.49(s,3H),1.31(s,6H),1.24(m,3H).1H NMR(301MHz, DMSO-d6)δ9.01(s,1H),8.57(d,J=1.6Hz,2H),8.33(s,1H),7.67(d,J=6.8Hz,2H),7.57 (s,1H), 7.38(s,1H), 4.45(s,2H), 3.83–3.75(m,2H), 3.49(s,3H), 1.31(s,6H), 1.24(m,3H).
实施例12：化合物P-12的制备Example 12: Preparation of Compound P-12
Figure PCTCN2021127847-appb-000048
Figure PCTCN2021127847-appb-000048
采用实施例11的方法，得到目标产物P-12。Using the method of Example 11, the target product P-12 was obtained.
LCMS:t R＝1.350min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝566.2[M+H] +. LCMS: t R = 1.350 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 566.2 [M+H] + .
HPLC:t R＝6.995min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =6.995min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(300MHz,DMSO-d 6)δ9.00(s,1H),8.56(s,2H),8.32(s,1H),7.65(d,J＝2.8Hz,2H),7.57(s,1H),7.38(s,1H),4.44(s,2H),3.78–3.71(m,2H),3.47(s,3H),1.71(dd,J＝15.2,7.6Hz,2H),1.30(s,6H),0.92(dt,J＝7.3,3.7Hz,3H). 1 H NMR (300MHz, DMSO-d 6 )δ9.00(s,1H),8.56(s,2H),8.32(s,1H),7.65(d,J=2.8Hz,2H),7.57(s, 1H), 7.38(s, 1H), 4.44(s, 2H), 3.78–3.71(m, 2H), 3.47(s, 3H), 1.71(dd, J=15.2, 7.6Hz, 2H), 1.30(s ,6H),0.92(dt,J=7.3,3.7Hz,3H).
实施例13：化合物P-13的制备Example 13: Preparation of Compound P-13
Figure PCTCN2021127847-appb-000049
Figure PCTCN2021127847-appb-000049
采用实施例11的方法，得到目标产物P-13，MS(ESI)m/z＝614.3[M+H] +. Using the method of Example 11, the target product P-13 was obtained, MS (ESI) m/z=614.3 [M+H] + .
实施例14：化合物P-14的制备Example 14: Preparation of Compound P-14
Figure PCTCN2021127847-appb-000050
Figure PCTCN2021127847-appb-000050
采用实施例11的方法，得到目标产物P-14，MS(ESI)m/z＝614.3[M+H] +. Using the method of Example 11, the target product P-14 was obtained, MS (ESI) m/z=614.3 [M+H] + .
实施例15：化合物P-15的制备Example 15: Preparation of compound P-15
Figure PCTCN2021127847-appb-000051
Figure PCTCN2021127847-appb-000051
采用实施例11的方法，得到目标产物P-15，MS(ESI)m/z＝538.4[M+H] +. Using the method of Example 11, the target product P-15 was obtained, MS (ESI) m/z=538.4 [M+H] + .
实施例16：化合物P-16的制备Example 16: Preparation of Compound P-16
Figure PCTCN2021127847-appb-000052
Figure PCTCN2021127847-appb-000052
将5-溴-2-氯嘧啶(化合物SM16,500mg，2.58mmol)，硫吗啉1,1-二氧化物(化合物SM17,384mg，2.84mmol)和碳酸钾(714mg，5.17mmol)在N,N-二甲基甲酰胺(10mL)中的混合物于130℃下搅拌2小时。此时，将其用冰水(100mL)稀释，并将混合物用乙酸乙酯(3×30mL)萃取。用饱和盐水(50mL)洗涤有机相，并用硫酸钠干燥。浓缩得到化合物12(600mg，2.05mmol，产率79.5％)，为白色固体。LCMS:t R＝1.185min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝292.0,294.0[M+H] +；将4-(5-溴嘧啶-2-基)硫代吗啉1,2-二氧化物(化合物12，350mg，1.20mmol)，4,4,5,5-四甲基-2-(4,4,5,5-四甲基-1,3,2-二氧杂硼硼烷-2-基)-1,3,2-二氧杂硼烷(化合物SM5,304mg，1.20mmol)，乙酸钾(235mg，2.40mmol)和1,1'-双(二苯基膦基)二茂铁二氯化钯(131mg,0.18mmol)在二氧六环中的混合物在氮气保护于80℃搅拌过夜。此时，将其用冰水(20mL)稀释，并用乙酸乙酯(3×20mL)萃取混合物。用饱和盐水(50mL)洗涤有机相，并用硫酸钠干燥。浓缩得到残留物。将其通过Combi-Flash纯化，用石油醚/乙酸乙酯(1∶1)洗脱，得到化合物13(300mg，0.88mmol，产率73.8％)，为棕色固体。LCMS:t R＝1.285min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝340.2[M+H] +；将(Z)-2-溴-3-(3-(3,3-二甲基-7-(三氟甲基)-2,3-二氢苯并呋喃-5-基)-1H-1,2,4-三唑-1-基)丙烯酰胺(化合物Int-2,80mg，0.18mmol)，4-(5-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)嘧啶-2-基)硫代吗啉1,1-二氧化物(化合物13，82mg，0.24mmol)，乙酸钾(46mg 0.46mmol) 和1,1'-双(二苯基膦基)二茂铁钯二氯化钯(20mg 0.028mmol)在二氧六环/水＝10：1(10mL)中的混合物在氮气保护下于80℃搅拌过夜。此时，将其用冰水(20mL)稀释，并将混合物用乙酸乙酯(3×10mL)萃取。用饱和盐水(20mL)洗涤有机相，并用硫酸钠干燥。浓缩得到残留物。通过用展开剂(二氯甲烷/甲醇＝15：1)的硅胶预制备板纯化得到粗品，并通过高效制备液相色谱纯化(流动相：乙腈/水＝35/65～70/30)得到化合物P-16(11.8mg，0.021mmol，产率11.3％)，为白色固体。 Combine 5-bromo-2-chloropyrimidine (compound SM16, 500 mg, 2.58 mmol), thiomorpholine 1,1-dioxide (compound SM17, 384 mg, 2.84 mmol) and potassium carbonate (714 mg, 5.17 mmol) under N, The mixture in N-dimethylformamide (10 mL) was stirred at 130°C for 2 hours. At this time, it was diluted with ice water (100 mL) and the mixture was extracted with ethyl acetate (3 x 30 mL). The organic phase was washed with saturated brine (50 mL) and dried over sodium sulfate. Concentration gave compound 12 (600 mg, 2.05 mmol, 79.5% yield) as a white solid. LCMS: t R =1.185 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm×50 mm), MS (ESI) m/z=292.0, 294.0 [M+H] + ; 4-(5-bromopyrimidine -2-yl)thiomorpholine 1,2-dioxide (compound 12, 350 mg, 1.20 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl) Methyl-1,3,2-dioxaboron-2-yl)-1,3,2-dioxaborane (compound SM5, 304 mg, 1.20 mmol), potassium acetate (235 mg, 2.40 mmol) A mixture of 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (131 mg, 0.18 mmol) in dioxane was stirred at 80°C overnight under nitrogen protection. At this time, it was diluted with ice water (20 mL) and the mixture was extracted with ethyl acetate (3 x 20 mL). The organic phase was washed with saturated brine (50 mL) and dried over sodium sulfate. Concentration gave a residue. It was purified by Combi-Flash eluting with petroleum ether/ethyl acetate (1:1) to give compound 13 (300 mg, 0.88 mmol, 73.8% yield) as a brown solid. LCMS: t R = 1.285 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm×50 mm), MS (ESI) m/z=340.2 [M+H] + ; (Z)-2-bromo-3 -(3-(3,3-Dimethyl-7-(trifluoromethyl)-2,3-dihydrobenzofuran-5-yl)-1H-1,2,4-triazole-1- yl)acrylamide (Compound Int-2, 80 mg, 0.18 mmol), 4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl) Pyrimidine-2-yl)thiomorpholine 1,1-dioxide (compound 13, 82 mg, 0.24 mmol), potassium acetate (46 mg 0.46 mmol) and 1,1'-bis(diphenylphosphino)diocene A mixture of iron palladium palladium dichloride (20 mg 0.028 mmol) in dioxane/water = 10:1 (10 mL) was stirred at 80° C. overnight under nitrogen protection. At this time, it was diluted with ice water (20 mL), and the mixture was extracted with ethyl acetate (3 x 10 mL). The organic phase was washed with saturated brine (20 mL) and dried over sodium sulfate. Concentration gave a residue. The crude product was purified by preparative silica gel plate purification with developing solvent (dichloromethane/methanol=15:1), and purified by high performance preparative liquid chromatography (mobile phase: acetonitrile/water=35/65～70/30) to obtain the compound P-16 (11.8 mg, 0.021 mmol, 11.3% yield) as a white solid.
LCMS:t R＝1.264min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝564.2[M+H] +. LCMS: t R = 1.264 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 564.2 [M+H] + .
HPLC:t R＝4.051min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =4.051min in Shimadzu 2010/2030, chromatography (XBRIDGE 2.1×50mm, 3.5um).
1H NMR(400MHz,DMSO)δ8.94(s,1H),8.34(s,2H),8.24(s,1H),7.77–7.69(m,2H),7.65–7.51(m,1H),7.28–7.13(m,1H),4.47(s,2H),4.26(s,4H),3.20–3.15(m,4H),1.33(s,6H). 1 H NMR(400MHz, DMSO)δ8.94(s,1H),8.34(s,2H),8.24(s,1H),7.77-7.69(m,2H),7.65-7.51(m,1H),7.28 –7.13(m,1H),4.47(s,2H),4.26(s,4H),3.20–3.15(m,4H),1.33(s,6H).
实施例17：化合物P-17的制备Example 17: Preparation of compound P-17
Figure PCTCN2021127847-appb-000053
Figure PCTCN2021127847-appb-000053
采用实施例16的方法，得到目标产物P-17，MS(ESI)m/z＝596.2[M+H] +. Using the method of Example 16, the target product P-17 was obtained, MS(ESI) m/z=596.2[M+H] + .
实施例18：化合物P-18的制备Example 18: Preparation of Compound P-18
Figure PCTCN2021127847-appb-000054
Figure PCTCN2021127847-appb-000054
采用实施例16的方法，得到目标产物P-18，MS(ESI)m/z＝532.3[M+H] +. Using the method of Example 16, the target product P-18 was obtained, MS (ESI) m/z=532.3 [M+H] + .
实施例19制备中间体Int-1-1和Int-2-1Example 19 Preparation of intermediates Int-1-1 and Int-2-1
Figure PCTCN2021127847-appb-000055
Figure PCTCN2021127847-appb-000055
合成路线如下：The synthetic route is as follows:
Figure PCTCN2021127847-appb-000056
Figure PCTCN2021127847-appb-000056
步骤1.1化合物1的制备Step 1.1 Preparation of Compound 1
Figure PCTCN2021127847-appb-000057
Figure PCTCN2021127847-appb-000057
把3-溴-1,2,4-三氮唑(SM1,4.44g，30.0mmol)溶解于30ml DMF中并加入三乙烯二胺(6.73g，60.0mmol)，在室温下搅拌30分钟。然后加入(2Z)-异丙基-3-碘丙-2-烯酸酯(7.92g，33.0mmol)，并在室温下搅拌最终混合物1h。将混合物倒入100ml冰水中，用乙酸乙酯(3x50ml)提取水相。合并的有机相用50ml盐水洗涤两次，用无水硫酸钠干燥，过滤，减压浓缩，得到粗产品。将粗品加入硅胶柱中，用(PE:EA＝5:1)洗脱，得到化合物1(6.25g，24.0mmol，产率80％)为类白色固体。3-Bromo-1,2,4-triazole (SM1, 4.44 g, 30.0 mmol) was dissolved in 30 ml of DMF and triethylenediamine (6.73 g, 60.0 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. Then (2Z)-isopropyl-3-iodoprop-2-enoate (7.92 g, 33.0 mmol) was added and the final mixture was stirred at room temperature for 1 h. The mixture was poured into 100ml ice water and the aqueous phase was extracted with ethyl acetate (3x50ml). The combined organic phases were washed twice with 50 ml of brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the crude product. The crude product was added to a silica gel column and eluted with (PE:EA=5:1) to give compound 1 (6.25 g, 24.0 mmol, 80% yield) as an off-white solid.
LCMS:t R＝1.222min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝259.8,261.7[M+H] +. LCMS: t R = 1.222 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 259.8, 261.7 [M+H] + .
步骤1.2化合物2的制备Step 1.2 Preparation of Compound 2
Figure PCTCN2021127847-appb-000058
Figure PCTCN2021127847-appb-000058
向化合物SM3(2.5g，10.4mmol)的氨水(50mL)溶液中加入碘化钾(5.2g，31.2mmol)和碘单质(2.6g，10.4mmol)的水(25mL)溶液。将混合物在室温下搅拌16小时。此时用浓盐酸将混合物调节至pH＝3并用EA(3×50mL)萃取。用饱和碳酸氢钠溶液(50mL)和食盐水(50mL)洗涤有机物。用硫酸钠干燥并浓缩，得到呈浅棕色油状的化合物4-溴-2-碘-6-(三氟甲基)苯酚(3.0g，8.2mmol，产率78.9％)。To a solution of compound SM3 (2.5 g, 10.4 mmol) in ammonia water (50 mL) was added a solution of potassium iodide (5.2 g, 31.2 mmol) and elemental iodine (2.6 g, 10.4 mmol) in water (25 mL). The mixture was stirred at room temperature for 16 hours. At this time the mixture was adjusted to pH=3 with concentrated hydrochloric acid and extracted with EA (3 x 50 mL). The organics were washed with saturated sodium bicarbonate solution (50 mL) and brine (50 mL). Drying over sodium sulfate and concentration gave compound 4-bromo-2-iodo-6-(trifluoromethyl)phenol (3.0 g, 8.2 mmol, 78.9% yield) as a light brown oil.
LCMS(ES,m/z):364.5[M-H]-,retention time 1.300min.LCMS(ES,m/z):364.5[M-H]-,retention time 1.300min.
步骤1.3化合物9的制备Step 1.3 Preparation of Compound 9
Figure PCTCN2021127847-appb-000059
Figure PCTCN2021127847-appb-000059
化合物2(3.1g，22.2mmol)在N,N-二甲基甲酰胺(40ml)的溶液中加入3-溴丙-1-烯(1.8g，14.8mmol)。 将混合物在室温下搅拌过夜。加入冰水(400mL)并将混合物用乙酸乙酯(3×100mL)萃取。有机相用饱和食盐水(100mL)洗涤并用无水硫酸钠干燥。浓缩，得到所需化合物9(2.5g，6.1mmol，粗产率82.4％)为浅棕色油状物。To a solution of compound 2 (3.1 g, 22.2 mmol) in N,N-dimethylformamide (40 ml) was added 3-bromoprop-1-ene (1.8 g, 14.8 mmol). The mixture was stirred at room temperature overnight. Ice water (400 mL) was added and the mixture was extracted with ethyl acetate (3 x 100 mL). The organic phase was washed with saturated brine (100 mL) and dried over anhydrous sodium sulfate. Concentration gave the desired compound 9 (2.5 g, 6.1 mmol, 82.4% crude yield) as a light brown oil.
LCMS(ES,m/z):No MS,retention time 1.300minLCMS(ES,m/z): No MS, retention time 1.300min
步骤1.4化合物10的制备Step 1.4 Preparation of Compound 10
Figure PCTCN2021127847-appb-000060
Figure PCTCN2021127847-appb-000060
将化合物9(20mg，0.12mmol)加入到甲苯(15mL)溶液中，混合物在115℃下搅拌。然后加入氟化钾(572mg，9.84mmol)的水溶液(1mL)和聚(甲基氢硅氧烷)(1.6g，7.38mmol)。将混合物在115℃下搅拌过夜。此时，将混合物用1摩尔/升氢氧化钠(100mL)淬灭并用乙酸乙酯(3×50mL)萃取。有机相用饱和氯化铵(100mL)，水(100mL)和饱和食盐水(100mL)洗涤，然后用无水硫酸钠干燥并浓缩，得到粗品。通过Combi快速柱层析(洗脱剂：石油醚/乙酸乙酯＝100:1)并且硅胶制备板(展开剂：石油醚/二氯甲烷＝12:1)纯化，得到化合物10(400mg，1.42mmol，产率57.9％)为无色油状物。Compound 9 (20 mg, 0.12 mmol) was added to a solution of toluene (15 mL), and the mixture was stirred at 115°C. Then potassium fluoride (572 mg, 9.84 mmol) in water (1 mL) and poly(methylhydrogensiloxane) (1.6 g, 7.38 mmol) were added. The mixture was stirred at 115°C overnight. At this time, the mixture was quenched with 1 mol/L sodium hydroxide (100 mL) and extracted with ethyl acetate (3 x 50 mL). The organic phase was washed with saturated ammonium chloride (100 mL), water (100 mL) and saturated brine (100 mL), then dried over anhydrous sodium sulfate and concentrated to obtain a crude product. Purification by Combi flash column chromatography (eluent: petroleum ether/ethyl acetate=100:1) and silica gel preparative plate (developing solvent: petroleum ether/dichloromethane=12:1) gave compound 10 (400 mg, 1.42 mmol, 57.9% yield) as a colorless oil.
1HNMR(400MHz,CDCl 3):δ7.48–7.44(m,1H),7.39(s,1H),4.83(t,J＝9.0Hz,1H),4.23(dd,J＝8.8,7.6Hz,1H),3.67–3.46(m,1H),1.35(d,J＝6.9Hz,3H). 1 H NMR (400 MHz, CDCl 3 ): δ 7.48-7.44 (m, 1H), 7.39 (s, 1H), 4.83 (t, J=9.0 Hz, 1H), 4.23 (dd, J=8.8, 7.6 Hz, 1H), 3.67–3.46 (m, 1H), 1.35 (d, J=6.9Hz, 3H).
步骤1.5化合物11的制备Step 1.5 Preparation of Compound 11
Figure PCTCN2021127847-appb-000061
Figure PCTCN2021127847-appb-000061
化合物10(650mg，2.31mmol)，联硼酸频那醇酯(880mg，3.47mmol)，乙酸钾(567mg，5.78mmol)和1，1-双(二-苯基磷基)二茂铁氯化钯(169mg，0.23mmol)的二氧六环溶液(15mL)在80℃下搅拌1小时。冷却至室温，加入水(30mL)并将混合物用乙酸乙酯(3×20mL)萃取。有机相用饱和食盐水(30mL)洗涤并用无水硫酸钠干燥。浓缩得到粗品。通过Combi快速柱层析(洗脱剂：石油 醚/乙酸乙酯＝100:1)洗脱纯化，得到化合物11(500mg，1.52mmol，产率66.0％)为无色油状物。Compound 10 (650 mg, 2.31 mmol), pinacol diboronate (880 mg, 3.47 mmol), potassium acetate (567 mg, 5.78 mmol) and 1,1-bis(di-phenylphosphonyl)ferrocene palladium chloride (169 mg, 0.23 mmol) in dioxane (15 mL) was stirred at 80 °C for 1 h. Cooled to room temperature, water (30 mL) was added and the mixture was extracted with ethyl acetate (3 x 20 mL). The organic phase was washed with saturated brine (30 mL) and dried over anhydrous sodium sulfate. Concentration gave crude product. Purification by Combi flash column chromatography (eluent: petroleum ether/ethyl acetate=100:1) gave compound 11 (500 mg, 1.52 mmol, yield 66.0%) as a colorless oil.
LCMS(ES,m/z):328.8[M+H] +,retention time 1.559min. LCMS(ES,m/z): 328.8[M+H] + , retention time 1.559min.
1H NMR(400MHz,CDCl 3):δ7.83(s,1H),7.74(s,1H),4.84(t,J＝9.0Hz,1H),4.24(dd,J＝8.8,7.5Hz,1H),3.63-3.51(m,1H),1.37(d,J＝6.9Hz,3H),1.34(d,J＝1.7Hz,12H). 1 H NMR (400 MHz, CDCl 3 ): δ 7.83 (s, 1H), 7.74 (s, 1H), 4.84 (t, J=9.0 Hz, 1H), 4.24 (dd, J=8.8, 7.5 Hz, 1H) ),3.63-3.51(m,1H),1.37(d,J=6.9Hz,3H),1.34(d,J=1.7Hz,12H).
步骤1.6化合物12的制备Step 1.6 Preparation of Compound 12
Figure PCTCN2021127847-appb-000062
Figure PCTCN2021127847-appb-000062
将化合物11(4.9g，15.0mmol)，((Z)-3-(3-溴-1H-1,2,4-三唑-1-基)丙烯酸异丙酯(3.9g，15.0mmol)，碳酸钾(6.21g，45.0mmol)和四三苯基膦钯(2.25g，1.95mmol)溶于二氧六环/水(63ml，20:1)中。将反应混合液在85℃氮气保护下搅拌10小时。浓缩得到粗产品。通过硅胶柱层析方法纯化，用石油醚/乙酸乙酯(5:1)洗脱，得到类白色固体状化合物12(3.8g，10.0mmol，产率66.7％)。Compound 11 (4.9 g, 15.0 mmol), ((Z)-3-(3-bromo-1H-1,2,4-triazol-1-yl)isopropyl acrylate (3.9 g, 15.0 mmol), Potassium carbonate (6.21 g, 45.0 mmol) and palladium tetrakistriphenylphosphine (2.25 g, 1.95 mmol) were dissolved in dioxane/water (63 ml, 20:1). The reaction mixture was heated at 85°C under nitrogen protection Stir for 10 hours. Concentrate to obtain crude product. Purify by silica gel column chromatography, eluting with petroleum ether/ethyl acetate (5:1), to give compound 12 (3.8 g, 10.0 mmol, 66.7% yield) as an off-white solid ).
LCMS:t R＝1.472min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝381.8[M+H] +. LCMS: t R = 1.472 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 381.8 [M+H] + .
1H NMR(400MHz,CDCl3):δ9.68(s,1H),8.18(s,1H),8.08(s,1H),7.25(d,J＝10.8Hz,1H),5.66(d,J＝10.8Hz,1H),5.15～5.10(m,1H),4.90(t,J＝8.8Hz,1H),4.29(t,J＝8.8Hz,1H),3.67～3.61(m,1H),1.42(d,J＝6.8Hz,3H),1.32(d,J＝6.0Hz,6H). 1 H NMR (400MHz, CDCl3): δ9.68(s,1H), 8.18(s,1H), 8.08(s,1H), 7.25(d,J=10.8Hz,1H), 5.66(d,J= 10.8Hz, 1H), 5.15～5.10(m, 1H), 4.90(t, J=8.8Hz, 1H), 4.29(t, J=8.8Hz, 1H), 3.67～3.61(m, 1H), 1.42( d,J=6.8Hz,3H),1.32(d,J=6.0Hz,6H).
步骤1.7化合物13的制备Step 1.7 Preparation of compound 13
Figure PCTCN2021127847-appb-000063
Figure PCTCN2021127847-appb-000063
将化合物12(4.9g，12.4mmol)溶解于二氯甲烷(80ml)，在0℃下缓慢滴加溴素(3.95g，24.8mmol)。混合物在室温下搅拌过夜。反应完倒入冰水(80mL)中，用二氯甲烷(2x 80mL)提取。合并有机相用亚硫酸氢钠水溶液(30mL)和饱和食盐水(2x 30mL)洗，无水硫酸钠干燥，减压浓缩，得到黄色油状的粗产品化合物13(4.87g，9.0mmol，粗产率90％)，直接用于下一步，没有经过任何纯化。Compound 12 (4.9 g, 12.4 mmol) was dissolved in dichloromethane (80 ml), and bromine (3.95 g, 24.8 mmol) was slowly added dropwise at 0°C. The mixture was stirred at room temperature overnight. After the reaction was completed, it was poured into ice water (80 mL) and extracted with dichloromethane (2×80 mL). The combined organic phases were washed with aqueous sodium bisulfite solution (30 mL) and saturated brine (2×30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a yellow oily crude product Compound 13 (4.87 g, 9.0 mmol, crude yield). 90%), used directly in the next step without any purification.
LCMS:t R＝1.478min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝539.4,541.4,543.4[M+H] +. LCMS: t R = 1.478 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 539.4, 541.4, 543.4 [M+H] + .
步骤1.8化合物14的制备Step 1.8 Preparation of Compound 14
Figure PCTCN2021127847-appb-000064
Figure PCTCN2021127847-appb-000064
将化合物13(4.87g，9.0mmol)溶解于四氢呋喃(50mL)在0度下加入三乙胺(1.82g，18mmol)。将混合物在室温下搅拌10h。加入30ml水稀释混合物，用乙酸乙酯3x 50ml提取水层，然后用50ml盐水萃洗有机相两次，用无水硫酸钠干燥，过滤，并减压浓缩，得到淡黄色固体的粗品通过硅胶柱纯化，用石油醚/乙酸乙酯(8:1)洗脱，得到化合物14(1.8g，3.9mmol，产率43.3％)为白色固体。Compound 13 (4.87 g, 9.0 mmol) was dissolved in tetrahydrofuran (50 mL) and triethylamine (1.82 g, 18 mmol) was added at 0 degrees. The mixture was stirred at room temperature for 10 h. 30 ml of water was added to dilute the mixture, the aqueous layer was extracted with ethyl acetate 3×50 ml, then the organic phase was washed twice with 50 ml of brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a pale yellow solid crude product that was passed through a silica gel column Purification, eluting with petroleum ether/ethyl acetate (8:1), afforded compound 14 (1.8 g, 3.9 mmol, 43.3% yield) as a white solid.
LCMS:t R＝1.490min MS(ESI)m/z＝459.6,461.6[M+H] +. LCMS: t R = 1.490 min MS (ESI) m/z = 459.6,461.6 [M+H] + .
1H NMR(400MHz,CDCl3):δ9.43(s,1H),8.75(s,1H),8.20(s,1H),8.09(s,1H),5.24～5.18(m,1H),4.92(t,J＝9.2Hz,1H),4.29(t,J＝8.8Hz,1H),3.69～3.63(m,1H),1.42(d,J＝6.8Hz,3H),1.32(d,J＝6.4Hz, 6H). 1 H NMR (400MHz, CDCl3): δ9.43(s,1H), 8.75(s,1H), 8.20(s,1H), 8.09(s,1H), 5.24～5.18(m,1H), 4.92( t,J＝9.2Hz,1H),4.29(t,J＝8.8Hz,1H),3.69～3.63(m,1H),1.42(d,J＝6.8Hz,3H),1.32(d,J＝6.4 Hz, 6H).
步骤1.9化合物Int-1-1的制备Step 1.9 Preparation of compound Int-1-1
Figure PCTCN2021127847-appb-000065
Figure PCTCN2021127847-appb-000065
化合物14(1200mg，2.6mmol)溶解于THF(25ml)中。冷却至0度缓慢滴加一水氢氧化锂(163.6mg，3.9mmol)和15mL水配成的溶液。反应混合液在0℃搅拌1h。反应液在真空下浓缩，剩余物用15ml水稀释。用甲基叔丁基醚2x 20ml反萃水相，然后用浓盐酸将水相调节到pH＝2，用乙酸乙酯3x30ml提取水层。用无水硫酸钠干燥，过滤，并减压浓缩，得到目标物Int-1-1(900mg，2.13mmol，产率81.9％)为白色固体。Compound 14 (1200 mg, 2.6 mmol) was dissolved in THF (25 ml). Cool to 0 degrees and slowly add a solution of lithium hydroxide monohydrate (163.6 mg, 3.9 mmol) and 15 mL of water dropwise. The reaction mixture was stirred at 0 °C for 1 h. The reaction solution was concentrated in vacuo and the residue was diluted with 15 ml of water. The aqueous phase was back-extracted with methyl tert-butyl ether 2×20 ml, then adjusted to pH=2 with concentrated hydrochloric acid, and the aqueous layer was extracted with ethyl acetate 3×30 ml. It was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target Int-1-1 (900 mg, 2.13 mmol, 81.9% yield) as a white solid.
LCMS:t R＝1.490min MS(ESI)m/z＝417.6,419.6[M+H] +. LCMS: t R = 1.490 min MS (ESI) m/z = 417.6, 419.6 [M+H] + .
1HNMR(400MHz,CDCl 3):δ13.95(brs,1H),9.43(s,1H),8.85(s,1H),8.15(s,1H),8.06(s,1H),4.98-4.91(m,1H),4.33(t,J＝9.2Hz,1H),,3.73-3.69(m,1H),1.36(d,J＝Hz,3H). 1 HNMR (400MHz, CDCl 3 ): δ13.95(brs,1H), 9.43(s,1H), 8.85(s,1H), 8.15(s,1H), 8.06(s,1H), 4.98-4.91( m,1H),4.33(t,J=9.2Hz,1H),,3.73-3.69(m,1H),1.36(d,J=Hz,3H).
步骤1.10中间体Int-2-1的制备Step 1.10 Preparation of intermediate Int-2-1
Figure PCTCN2021127847-appb-000066
Figure PCTCN2021127847-appb-000066
在化合物12(600mg，1.573mmol)的四氢呋喃(10mL)溶液中加入1M的氢氧化锂(10mL)。将混合物在室温下搅拌2.5小时。此时，用HCl调节混合物pH＝3，并将混合物用乙酸乙酯(3×15mL)萃取。有机相用饱和盐水(30mL)洗涤并用无水硫酸钠干燥。浓缩得到化合物Int-2-1(500mg，1.47mmol，产率93.7％)，浅黄色固体。To a solution of compound 12 (600 mg, 1.573 mmol) in tetrahydrofuran (10 mL) was added 1 M lithium hydroxide (10 mL). The mixture was stirred at room temperature for 2.5 hours. At this time, the mixture was adjusted to pH=3 with HCl, and the mixture was extracted with ethyl acetate (3 x 15 mL). The organic phase was washed with saturated brine (30 mL) and dried over anhydrous sodium sulfate. Concentration gave compound Int-2-1 (500 mg, 1.47 mmol, 93.7% yield) as a pale yellow solid.
LCMS:t R＝1.293min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝340.7[M+H] +. LCMS: t R = 1.293 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 340.7 [M+H] + .
实施例20：化合物P-19的制备Example 20: Preparation of Compound P-19
Figure PCTCN2021127847-appb-000067
Figure PCTCN2021127847-appb-000067
将化合物Int-1-1(83.6mg，0.2mmol)溶解在30mL THF中。在0度下添加氮甲基吗啡啉(30.3mg，0.3mmol)和氯甲酸异丙酯(49mg，0.4mmol)。反应混合液在0度搅拌1小时后加入三氟乙胺(198mg，2.0mmol)，之后继续搅拌1小时。所得到的混合物在真空下浓缩，剩余物用20ml冰水稀释，EA(3x30ml)萃取。合并有机相用是盐水洗，硫酸钠干燥，在真空下浓缩得到粗产品。以石油醚/乙酸乙酯＝1/2为展开剂，通过制备薄层色谱法对其进行纯化，得到目标中间体15(50mg，0.1mmol，产率50.1％)为白色固体。Compound Int-1-1 (83.6 mg, 0.2 mmol) was dissolved in 30 mL of THF. Nitromethylmorpholine (30.3 mg, 0.3 mmol) and isopropyl chloroformate (49 mg, 0.4 mmol) were added at 0 degrees. The reaction mixture was stirred at 0°C for 1 hour, then trifluoroethylamine (198 mg, 2.0 mmol) was added, and stirring was continued for 1 hour. The resulting mixture was concentrated in vacuo, the residue was diluted with 20ml ice water and extracted with EA (3x30ml). The combined organic phases were washed with brine, dried over sodium sulfate, and concentrated in vacuo to give the crude product. Using petroleum ether/ethyl acetate=1/2 as a developing solvent, it was purified by preparative thin layer chromatography to obtain the target intermediate 15 (50 mg, 0.1 mmol, yield 50.1%) as a white solid.
LCMS:t R＝1.349min MS(ESI)m/z＝498.5,500.5[M+H] +. LCMS: t R = 1.349 min MS (ESI) m/z = 498.5,500.5 [M+H] + .
Figure PCTCN2021127847-appb-000068
Figure PCTCN2021127847-appb-000068
将化合物15(50mg，0.1mmol)、5-硼酸嘧啶(24.8mg，0.2mmol)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(14.6mg，0.022mmol)和乙酸钾(29.5mg，0.3mmol)加入到10/1mL二氧六环/H 2O溶液中。在90℃氮气保护下搅拌10小时。将反应液浓缩。以纯PE/EA(1/2)为展开剂，采用制备TLC纯化得到粗品，用制备HPLC(流动相：ACN/H 2O:55/45～40/60)纯化得到目标产物P-19(20mg,0.04mmol,产率40％)白色固体。 Compound 15 (50 mg, 0.1 mmol), 5-boronic acid pyrimidine (24.8 mg, 0.2 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloride palladium (14.6 mg, 0.022 mmol) ) and potassium acetate (29.5 mg, 0.3 mmol) were added to a 10/1 mL dioxane/ H2O solution. Stir under nitrogen protection at 90°C for 10 hours. The reaction solution was concentrated. Using pure PE/EA (1/2) as developing solvent, the crude product was purified by preparative TLC, and purified by preparative HPLC (mobile phase: ACN/H 2 O: 55/45～40/60) to obtain the target product P-19 (20 mg) , 0.04 mmol, 40% yield) white solid.
LCMS:t R＝1.262min,MS(ESI)m/z＝498.6[M+H] +. LCMS: t R = 1.262 min, MS (ESI) m/z = 498.6 [M+H] + .
HPLC:t R＝6.459min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =6.459min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(400MHz,DMSO-d 6):δ9.25(s,1H),9.07(s,1H),8.72(s,2H),8.42(s,1H),8.41(t,J＝6.0Hz,1H),7.66(s,1H),7.54(s,1H),4.91(t,J＝8.8Hz,1H),4.27(t,J＝8.0Hz,1H),4.03～3.95(m,2H),3.67～3.62(m,1H),1.31(d,J＝7.2Hz,3H). 1 H NMR (400MHz, DMSO-d 6 ): δ 9.25(s, 1H), 9.07(s, 1H), 8.72(s, 2H), 8.42(s, 1H), 8.41(t, J=6.0Hz ,1H),7.66(s,1H),7.54(s,1H),4.91(t,J＝8.8Hz,1H),4.27(t,J＝8.0Hz,1H),4.03～3.95(m,2H) ,3.67～3.62(m,1H),1.31(d,J=7.2Hz,3H).
实施例21：化合物P-20的制备Example 21: Preparation of Compound P-20
Figure PCTCN2021127847-appb-000069
Figure PCTCN2021127847-appb-000069
将化合物Int-1-1(80mg,0.19mmol)和二甲胺(25.87mg,0.57mmol)溶于DMF(10mL)中。常温下加入HATU(87.29mg,0.23mmol)和DIEA(0.09mL,0.57mmol)。室温下搅拌2h，加入H 2O(20mL)并用EA(20mL*2)萃取。有机层用盐水(20ml*3)冲洗，并在真空下浓缩，经TLC(EA)纯化，得到目标中间体16(60mg，0.13mmol,产率68.4％)，黄色油状。 Compound Int-1-1 (80 mg, 0.19 mmol) and dimethylamine (25.87 mg, 0.57 mmol) were dissolved in DMF (10 mL). HATU (87.29 mg, 0.23 mmol) and DIEA (0.09 mL, 0.57 mmol) were added at room temperature. Stir at room temperature for 2 h, add H2O (20 mL) and extract with EA (20 mL*2). The organic layer was washed with brine (20 ml*3) and concentrated in vacuo, purified by TLC (EA) to give the target intermediate 16 (60 mg, 0.13 mmol, 68.4% yield) as a yellow oil.
LCMS:t R＝1.271min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝444.6,446.6[M+H] +. LCMS: t R = 1.271 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 444.6, 446.6 [M+H] + .
Figure PCTCN2021127847-appb-000070
Figure PCTCN2021127847-appb-000070
将化合物15(60mg,0.13mmol)和嘧啶-5-基硼酸(41.76mg,0.34mmol)溶于二氧六环/H2O(10mL/1mL)中，常温下加入Pd(dppf)Cl 2-DCM(22mg,0.027mmol)和CH 3COOK(40mg,0.40mmol)。88℃搅拌过夜，经硅藻土过滤，滤液真空浓缩。残留物经HPLC(流动相：ACN/H2O＝30/70～80/20)纯化，得到目标产物P-20(13.4mg,产率20.18％)，灰白色固体。 Compound 15 (60 mg, 0.13 mmol) and pyrimidin-5-ylboronic acid (41.76 mg, 0.34 mmol) were dissolved in dioxane/H2O (10 mL/1 mL), and Pd(dppf)Cl 2 -DCM ( 22 mg, 0.027 mmol) and CH3COOK (40 mg, 0.40 mmol). Stir overnight at 88°C, filter through celite, and concentrate the filtrate in vacuo. The residue was purified by HPLC (mobile phase: ACN/H2O=30/70～80/20) to obtain the target product P-20 (13.4 mg, yield 20.18%) as an off-white solid.
LCMS:t R＝1.155min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS (ESI)m/z＝444.8[M+H] +. LCMS: t R = 1.155 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 444.8 [M+H] + .
HPLC:t R＝5.515min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =5.515min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(300MHz,CDCl 3):δ9.24(s,1H),8.91(s,2H),8.38(s,1H),8.15(s,1H),8.06(s,1H),7.34(s,1H),4.95～4.89(m,1H),4.31～4.28(m,1H),3.66(brs,1H),3.21(s,3H),2.96(s,3H),1.42(d,J＝6Hz,3H). 1 H NMR (300MHz, CDCl 3 ): δ9.24(s,1H), 8.91(s,2H), 8.38(s,1H), 8.15(s,1H), 8.06(s,1H), 7.34(s ,1H),4.95～4.89(m,1H),4.31～4.28(m,1H),3.66(brs,1H),3.21(s,3H),2.96(s,3H),1.42(d,J=6Hz , 3H).
实施例22：化合物P-21的制备Example 22: Preparation of Compound P-21
Figure PCTCN2021127847-appb-000071
Figure PCTCN2021127847-appb-000071
将化合物Int-1-1(60mg,0.14mmol)和异丙胺(25.45mg,0.43mmol)溶于DMF(10mL)中。常温下加入HATU(65.48mg,0.17mmol)和DIEA(0.07mL,0.43mmol)。室温下搅拌2h，加入H2O(20mL)并用EA(20mL*2)萃取。有机层用盐水(20ml*3)冲洗，并在真空下浓缩，经TLC(EA)纯化，得到目标中间体17(55mg，产率85.71％)，黄色油状。Compound Int-1-1 (60 mg, 0.14 mmol) and isopropylamine (25.45 mg, 0.43 mmol) were dissolved in DMF (10 mL). HATU (65.48 mg, 0.17 mmol) and DIEA (0.07 mL, 0.43 mmol) were added at room temperature. Stir at room temperature for 2 h, add H2O (20 mL) and extract with EA (20 mL*2). The organic layer was washed with brine (20 ml*3) and concentrated in vacuo, purified by TLC (EA) to give the target intermediate 17 (55 mg, 85.71% yield) as a yellow oil.
LCMS:t R＝1.408min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝458.6,460.7[M+H] +. LCMS: t R = 1.408 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 458.6, 460.7 [M+H] + .
Figure PCTCN2021127847-appb-000072
Figure PCTCN2021127847-appb-000072
将化合物17(55mg,0.12mmol)和嘧啶-5-基硼酸(37.11mg,0.3mmol)溶于二氧六环/H 2O(10mL/1mL)中，常温下加入Pd(dppf)Cl 2(19.55mg,0.024mmol)和CH 3COOK(35mg,0.36mmol)。90℃搅拌过夜，经硅藻土过滤，滤液真空浓缩。残留物经HPLC(流动相：ACN/H 2O＝30/70～80/20)纯化，得到目标产物P-21(12.3mg,产率21.85％)，白色固体。 Compound 17 (55 mg, 0.12 mmol) and pyrimidin-5-ylboronic acid (37.11 mg, 0.3 mmol) were dissolved in dioxane/H 2 O (10 mL/1 mL), and Pd(dppf)Cl 2 ( 19.55 mg, 0.024 mmol) and CH3COOK (35 mg, 0.36 mmol). Stir overnight at 90°C, filter through celite, and concentrate the filtrate in vacuo. The residue was purified by HPLC (mobile phase: ACN/H 2 O=30/70～80/20) to obtain the target product P-21 (12.3 mg, yield 21.85%) as a white solid.
LCMS:t R＝1.233min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS (ESI)m/z＝458.8[M+H] +. LCMS: t R = 1.233 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 458.8 [M+H] + .
HPLC:t R＝5.922min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =5.922min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(300MHz,CDCl3):δ9.36(s,1H),8.74(s,2H),8.32(d,J＝12Hz,2H),7.66(d,J＝12Hz,2H),5.20(d,J＝9Hz,1H),4.88(t,J＝9Hz,1H),4.28～4.19(m,2H),3.62～3.57(m,1H),1.36(d,J＝6Hz,3H),1.16(d,J＝6Hz,6H). 1 H NMR (300MHz, CDCl3): δ9.36(s, 1H), 8.74(s, 2H), 8.32(d, J=12Hz, 2H), 7.66(d, J=12Hz, 2H), 5.20(d ,J=9Hz,1H),4.88(t,J=9Hz,1H),4.28～4.19(m,2H),3.62～3.57(m,1H),1.36(d,J=6Hz,3H),1.16( d,J=6Hz,6H).
实施例23：化合物P-22的制备Example 23: Preparation of Compound P-22
Figure PCTCN2021127847-appb-000073
Figure PCTCN2021127847-appb-000073
将化合物Int-1-1(200mg,0.478mmol)溶于四氢呋喃(10mL)中，0℃下，加入氯(异丙氧基)甲酮(117.23mg,0.957mmol)和N-甲基吗啡啉(72.57mg,0.717mmol)。反应混合物在0℃下搅拌1h，然后加入吡咯烷(340.17mg,4.783mmol)，在0℃下继续搅拌1h。反应完，倒入水(10mL)中，乙酸乙酯(2×20mL)萃取。有机层用盐水(20mL)洗涤，无水硫酸镁干燥，过滤，减压浓缩。采用flash色谱法(洗脱剂：乙酸乙酯/石油醚＝4/5)进行分离纯化，得到目标中间体18(80mg，产率35.6％)，浅黄色固体。Compound Int-1-1 (200 mg, 0.478 mmol) was dissolved in tetrahydrofuran (10 mL), and at 0 °C, chloro(isopropoxy) ketone (117.23 mg, 0.957 mmol) and N-methylmorpholine ( 72.57 mg, 0.717 mmol). The reaction mixture was stirred at 0 °C for 1 h, then pyrrolidine (340.17 mg, 4.783 mmol) was added and stirring was continued at 0 °C for 1 h. After the reaction was completed, poured into water (10 mL) and extracted with ethyl acetate (2×20 mL). The organic layer was washed with brine (20 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. Using flash chromatography (eluent: ethyl acetate/petroleum ether=4/5) for separation and purification, the target intermediate 18 (80 mg, yield 35.6%) was obtained as a pale yellow solid.
LCMS:tR＝1.321min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝470.7,472.6[M+H] +. LCMS: tR=1.321min in Shimadzu LC20, chromatography (HALO C18 2.7μm, 4.6mm×50mm), MS(ESI) m/z=470.7,472.6[M+H] + .
Figure PCTCN2021127847-appb-000074
Figure PCTCN2021127847-appb-000074
将化合物18(80mg,0.17mmol)、嘧啶-5-基硼二醇(42.08mg,0.34mmol)和乙酸钾(50mg,0.509 mmol)溶于二氧六环/水(11mL,v/v＝10:1)中，加入[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(13.86mg,0.017mmol)。反应混合物在氮气保护下80℃搅拌10h，然后过滤，减压浓缩。采用prep-TLC(洗脱剂：乙酸乙酯)和prep-HPLC进行分离纯化，得到目标产物P-22(18.4mg，产率22.9％)，白色固体。Compound 18 (80 mg, 0.17 mmol), pyrimidin-5-ylborondiol (42.08 mg, 0.34 mmol) and potassium acetate (50 mg, 0.509 mmol) were dissolved in dioxane/water (11 mL, v/v=10 : 1), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (13.86 mg, 0.017 mmol) was added. The reaction mixture was stirred at 80°C for 10 h under nitrogen protection, then filtered and concentrated under reduced pressure. Using prep-TLC (eluent: ethyl acetate) and prep-HPLC for separation and purification, the target product P-22 (18.4 mg, yield 22.9%) was obtained as a white solid.
LCMS:tR＝1.207min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝470.7[M+H] +. LCMS: tR=1.207min in Shimadzu LC20, chromatography (HALO C18 2.7μm, 4.6mm×50mm), MS(ESI) m/z=470.7[M+H] + .
HPLC:tR＝4.157min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).HPLC:tR=4.157min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(301MHz,DMSO-d6):δ9.19(d,J＝8.5Hz,1H),8.94(s,1H),8.89–8.78(m,2H),8.10–7.91(m,2H),7.67(d,J＝31.4Hz,1H),4.93(dd,J＝21.7,9.0Hz,1H),4.37–4.19(m,1H),3.68–3.57(m,2H),3.45(dd,J＝9.8,6.2,2.5Hz,1H),3.32–3.11(m,2H),1.83(dd,J＝16.3,9.3,5.1Hz,4H),1.33(dd,J＝15.0,6.9Hz,3H). 1 H NMR (301MHz, DMSO-d6): δ9.19 (d, J=8.5Hz, 1H), 8.94 (s, 1H), 8.89–8.78 (m, 2H), 8.10–7.91 (m, 2H), 7.67 (d, J=31.4Hz, 1H), 4.93 (dd, J=21.7, 9.0Hz, 1H), 4.37–4.19 (m, 1H), 3.68–3.57 (m, 2H), 3.45 (dd, J= 9.8, 6.2, 2.5Hz, 1H), 3.32–3.11 (m, 2H), 1.83 (dd, J=16.3, 9.3, 5.1Hz, 4H), 1.33 (dd, J=15.0, 6.9Hz, 3H).
实施例24：化合物P-23的制备Example 24: Preparation of Compound P-23
Figure PCTCN2021127847-appb-000075
Figure PCTCN2021127847-appb-000075
将化合物Int-1-1(60mg,0.144mmol)溶于四氢呋喃(10mL)中，0℃下，加入氯(异丙氧基)甲酮(35.17mg,0.287mmol)和N-甲基吗啡啉(21.77mg,0.215mmol)。反应混合物在0℃下搅拌1h，然后加入环戊基胺(122.19mg,1.435mmol)，在0℃下继续搅拌1h。反应完，倒入水(10mL)中，乙酸乙酯(2×20mL)萃取。有机层用盐水(20mL)洗涤，无水硫酸镁干燥，过滤，减压浓缩，得到目标中间体19(60mg，产率86.13％)，浅黄色固体。Compound Int-1-1 (60 mg, 0.144 mmol) was dissolved in tetrahydrofuran (10 mL), chloro(isopropoxy) ketone (35.17 mg, 0.287 mmol) and N-methylmorpholine ( 21.77 mg, 0.215 mmol). The reaction mixture was stirred at 0 °C for 1 h, then cyclopentylamine (122.19 mg, 1.435 mmol) was added and stirring was continued at 0 °C for 1 h. After the reaction was completed, poured into water (10 mL) and extracted with ethyl acetate (2×20 mL). The organic layer was washed with brine (20 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to give the target intermediate 19 (60 mg, yield 86.13%) as a pale yellow solid.
LCMS:tR＝1.498min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝485.1,487.1[M+H] +. LCMS: tR=1.498min in Shimadzu LC20, chromatography (HALO C18 2.7μm, 4.6mm×50mm), MS(ESI) m/z=485.1,487.1[M+H] + .
Figure PCTCN2021127847-appb-000076
Figure PCTCN2021127847-appb-000076
将化合物19(60mg,0.124mmol)、嘧啶-5-基硼二醇(22.97mg,0.185mmol)和乙酸钾(24.26mg,0.247mmol)溶于二氧六环/水(11mL,v/v＝10:1)中，加入[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(10.12mg,0.0123mmol)。反应混合物在氮气保护下80℃搅拌10h，然后过滤，减压浓缩。采用flash色谱法(洗脱剂：乙酸乙酯)和prep-HPLC进行分离纯化，得到目标产物P-23(6.1mg，纯度98.63％，产率10％)，白色固体。Compound 19 (60 mg, 0.124 mmol), pyrimidin-5-ylborondiol (22.97 mg, 0.185 mmol) and potassium acetate (24.26 mg, 0.247 mmol) were dissolved in dioxane/water (11 mL, v/v= 10:1), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (10.12 mg, 0.0123 mmol) was added. The reaction mixture was stirred at 80°C for 10 h under nitrogen protection, then filtered and concentrated under reduced pressure. Using flash chromatography (eluent: ethyl acetate) and prep-HPLC for separation and purification, the target product P-23 (6.1 mg, purity 98.63%, yield 10%) was obtained as a white solid.
LCMS:t R＝1.290min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝485.1[M+H] +. LCMS: t R = 1.290 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 485.1 [M+H] + .
HPLC:t R＝6.609min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =6.609min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(300MHz,CD 3OD):δ9.21(s,1H),8.76(s,3H),8.24(s,1H),7.69(d,J＝6.2Hz,2H),4.86(d,J＝9.1Hz,1H),4.30–4.23(m,2H),3.63(dd,J＝15.4,7.4Hz,1H),2.00(d,J＝15.3Hz,2H),1.76–1.67(m,2H),1.57(dd,J＝23.5,8.8Hz,4H),1.37(d,J＝6.9Hz,3H). 1 H NMR (300 MHz, CD 3 OD): δ 9.21 (s, 1H), 8.76 (s, 3H), 8.24 (s, 1H), 7.69 (d, J=6.2 Hz, 2H), 4.86 (d, J=9.1Hz, 1H), 4.30–4.23 (m, 2H), 3.63 (dd, J=15.4, 7.4Hz, 1H), 2.00 (d, J=15.3Hz, 2H), 1.76–1.67 (m, 2H) ),1.57(dd,J＝23.5,8.8Hz,4H),1.37(d,J＝6.9Hz,3H).
实施例25：化合物P-24的制备Example 25: Preparation of Compound P-24
Figure PCTCN2021127847-appb-000077
Figure PCTCN2021127847-appb-000077
将化合物Int-1-1(Z)-2-溴-3-(3-甲基-7-(三氟甲基)-2,3-二氢苯并呋喃-5-基)-1H-1,2,4-三唑-1-基)丙烯酸(80mg,0.191mmol)溶于四氢呋喃(10mL)中，0℃下，加入氯(异丙氧基)甲酮(46.89mg,0.383mmol)和N-甲基吗啡啉(29.02mg,0.287mmol)。反应混合物在0℃下搅拌1h，然后加入环丙基胺(109.22mg,1.913mmol)，在0℃下继续搅拌1h。反应完，倒入水(10mL)中，乙酸乙酯(2×20mL)萃取。 有机层用盐水(20mL)洗涤，无水硫酸镁干燥，过滤，减压浓缩，得到目标中间体20(70mg，产率80.0％)，浅黄色固体。Compound Int-1-1(Z)-2-bromo-3-(3-methyl-7-(trifluoromethyl)-2,3-dihydrobenzofuran-5-yl)-1H-1 , 2,4-triazol-1-yl)acrylic acid (80 mg, 0.191 mmol) was dissolved in tetrahydrofuran (10 mL), and at 0°C, chloro(isopropoxy) ketone (46.89 mg, 0.383 mmol) and N was added. - Methylmorpholine (29.02 mg, 0.287 mmol). The reaction mixture was stirred at 0 °C for 1 h, then cyclopropylamine (109.22 mg, 1.913 mmol) was added and stirring was continued at 0 °C for 1 h. After the reaction was completed, poured into water (10 mL) and extracted with ethyl acetate (2×20 mL). The organic layer was washed with brine (20 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to give the target intermediate 20 (70 mg, yield 80.0%) as a pale yellow solid.
LCMS:tR＝1.328min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝456.6,458.5[M+H] +. LCMS: tR=1.328min in Shimadzu LC20, chromatography (HALO C18 2.7μm, 4.6mm×50mm), MS(ESI) m/z=456.6,458.5[M+H] + .
Figure PCTCN2021127847-appb-000078
Figure PCTCN2021127847-appb-000078
将化合物20(70mg,0.153mmol)、嘧啶-5-基硼二醇(28.46mg,0.23mmol)和乙酸钾(30.05mg,0.306mmol)溶于二氧六环/水(11mL,v/v＝10:1)中，加入[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(12.53mg,0.0153mmol)。反应混合物在氮气保护下80℃搅拌10h，然后过滤，减压浓缩。采用flash色谱法(洗脱剂：乙酸乙酯)和prep-HPLC进行分离纯化，得到目标产物P-24(21mg，纯度97.53％，产率29.3％)，白色固体。Compound 20 (70 mg, 0.153 mmol), pyrimidin-5-ylborodiol (28.46 mg, 0.23 mmol) and potassium acetate (30.05 mg, 0.306 mmol) were dissolved in dioxane/water (11 mL, v/v= 10:1), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (12.53 mg, 0.0153 mmol) was added. The reaction mixture was stirred at 80°C for 10 h under nitrogen protection, then filtered and concentrated under reduced pressure. Using flash chromatography (eluent: ethyl acetate) and prep-HPLC for separation and purification, the target product P-24 (21 mg, purity 97.53%, yield 29.3%) was obtained as a white solid.
LCMS:t R＝1.176min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝456.8[M+H] +. LCMS: t R = 1.176 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 456.8 [M+H] + .
HPLC:t R＝4.102min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =4.102min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(300MHz,DMSO-d 6):δ9.20(s,1H),9.01(s,1H),8.67(s,2H),8.31(s,1H),7.92(d,J＝4.1Hz,1H),7.66(s,1H),7.54(s,1H),4.90(t,J＝9.1Hz,1H),4.24(t,J＝8.3Hz,1H),3.65(dt,J＝16.9,8.4Hz,1H),2.80(dq,J＝11.5,3.8Hz,1H),1.30(d,J＝6.9Hz,3H),0.70–0.62(m,2H),0.56–0.48(m,2H). 1 H NMR (300MHz, DMSO-d 6 ): δ 9.20(s, 1H), 9.01(s, 1H), 8.67(s, 2H), 8.31(s, 1H), 7.92(d, J=4.1Hz ,1H),7.66(s,1H),7.54(s,1H),4.90(t,J＝9.1Hz,1H),4.24(t,J＝8.3Hz,1H),3.65(dt,J＝16.9, 8.4Hz, 1H), 2.80 (dq, J=11.5, 3.8Hz, 1H), 1.30 (d, J=6.9Hz, 3H), 0.70–0.62 (m, 2H), 0.56–0.48 (m, 2H).
实施例26：化合物P-25的制备Example 26: Preparation of Compound P-25
Figure PCTCN2021127847-appb-000079
Figure PCTCN2021127847-appb-000079
将化合物Int-1-1(1000mg，2.31mmol)溶解在30mL THF中。在0度下添加氮甲基吗啡啉(350mg，3.47mmol)和氯甲酸异丙酯(196mg，1.6mmol)。反应混合液在0度搅拌1小时后加入乙胺/四氢呋喃溶液(1mL，7M)，之后继续搅拌1小时。所得到的混合物在真空下浓缩，剩余物用30ml冰水稀释，沉淀析出，收集沉淀并用水洗(5mL)。白色固体即为目标中间体21(84.2mg，产率9％)Compound Int-1-1 (1000 mg, 2.31 mmol) was dissolved in 30 mL of THF. Nitromethylmorpholine (350 mg, 3.47 mmol) and isopropyl chloroformate (196 mg, 1.6 mmol) were added at 0 degrees. The reaction mixture was stirred at 0 degrees for 1 hour, then ethylamine/tetrahydrofuran solution (1 mL, 7M) was added, and stirring was continued for 1 hour. The resulting mixture was concentrated in vacuo, the residue was diluted with 30 ml of ice water and a precipitate was precipitated which was collected and washed with water (5 mL). The white solid is the target intermediate 21 (84.2 mg, yield 9%)
LCMS:t R＝1.328min MS(ESI)m/z＝444.6,446.7[M+H] +. LCMS: t R = 1.328 min MS (ESI) m/z = 444.6, 446.7 [M+H] + .
Figure PCTCN2021127847-appb-000080
Figure PCTCN2021127847-appb-000080
将化合物21(66mg，0.148mmol)、5-硼酸嘧啶(27.5mg，0.22mmol)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(18.12mg，0.022mmol)和乙酸钾(43.57mg，0.444mmol)加入到10/1mL二氧六环/H2O溶液中。在90℃氮气保护下搅拌10小时。将反应液浓缩。以纯EA为展开剂，采用制备TLC纯化得到粗品，用制备HPLC(流动相：ACN/H2O:60/40～40/60)纯化得到目标产物P-25(12mg,产率18.3％)白色固体。Compound 21 (66 mg, 0.148 mmol), 5-boronic acid pyrimidine (27.5 mg, 0.22 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloride palladium (18.12 mg, 0.022 mmol) ) and potassium acetate (43.57 mg, 0.444 mmol) were added to a 10/1 mL dioxane/H2O solution. Stir under nitrogen protection at 90°C for 10 hours. The reaction solution was concentrated. Using pure EA as the developing solvent, the crude product was purified by preparative TLC, and purified by preparative HPLC (mobile phase: ACN/H2O: 60/40～40/60) to obtain the target product P-25 (12 mg, yield 18.3%) as a white solid.
LCMS:t R＝1.209min,MS(ESI)m/z＝444.7[M+H] +. LCMS: t R = 1.209 min, MS (ESI) m/z = 444.7 [M+H] + .
HPLC:t R＝3.586min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =3.586min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(400MHz,MeOD):δ9.23(s,1H),8.78(s,1H),8.76(s,2H),8.32(s,1H),7.67(d,J＝11.2Hz,2H),4.87(t,J＝9.2Hz,1H),4.26(t,J＝8.8Hz,1H),3.65～3.59(m,1H),3.33(t,J＝7.2Hz,2H),1.42(d,J＝6.8Hz,3H),1.36(d,J＝6.8Hz,3H),1.16(t,J＝7.2Hz,3H). 1 H NMR (400MHz, MeOD): δ 9.23(s, 1H), 8.78(s, 1H), 8.76(s, 2H), 8.32(s, 1H), 7.67(d, J=11.2Hz, 2H) ,4.87(t,J＝9.2Hz,1H),4.26(t,J＝8.8Hz,1H),3.65～3.59(m,1H),3.33(t,J＝7.2Hz,2H),1.42(d, J=6.8Hz, 3H), 1.36 (d, J=6.8Hz, 3H), 1.16 (t, J=7.2Hz, 3H).
实施例27：化合物P-26的制备Example 27: Preparation of Compound P-26
Figure PCTCN2021127847-appb-000081
Figure PCTCN2021127847-appb-000081
将化合物Int-1-1(83mg，0.2mmol)甲胺盐酸盐(27mg，0.4mmol)溶于DMF(3mL)中并加入HATU(91.3mg，0.24mmol)和二异丙基乙胺(129mg，1.0mmol)。反应液在室温下搅拌10小时。用饱和NH4Cl(30ml)稀释反应液并用EA(3x30ml)萃取。溶剂用硫酸钠干燥，在真空下浓缩得到粗产品。以石油醚/乙酸乙酯＝1/1为展开剂，通过制备薄层色谱法对其进行纯化，得到目标中间体22(70mg，产率80.4％)为白色固体。Compound Int-1-1 (83 mg, 0.2 mmol) methylamine hydrochloride (27 mg, 0.4 mmol) was dissolved in DMF (3 mL) and HATU (91.3 mg, 0.24 mmol) and diisopropylethylamine (129 mg) were added , 1.0 mmol). The reaction solution was stirred at room temperature for 10 hours. The reaction was diluted with saturated NH4Cl (30ml) and extracted with EA (3x30ml). The solvent was dried over sodium sulfate and concentrated in vacuo to give the crude product. Using petroleum ether/ethyl acetate=1/1 as a developing solvent, it was purified by preparative thin layer chromatography to obtain the target intermediate 22 (70 mg, yield 80.4%) as a white solid.
LCMS:t R＝1.294min MS(ESI)m/z＝430.6,432.6[M+H] +. LCMS: t R = 1.294 min MS (ESI) m/z = 430.6, 432.6 [M+H] + .
1H NMR(400MHz,MeOD):δ9.26(s,1H),8.65(s,1H),8.15(s,1H),8.13(s,1H),4.90(t,J＝8.8Hz,1H),4.30(t,J＝8.8Hz,1H),3.74～3.66(m,1H),2.89(s,3H),1.30(d,J＝6.8Hz,3H). 1 H NMR (400MHz, MeOD): δ 9.26(s, 1H), 8.65(s, 1H), 8.15(s, 1H), 8.13(s, 1H), 4.90(t, J=8.8Hz, 1H) ,4.30(t,J＝8.8Hz,1H),3.74～3.66(m,1H),2.89(s,3H),1.30(d,J＝6.8Hz,3H).
Figure PCTCN2021127847-appb-000082
Figure PCTCN2021127847-appb-000082
将化合物22(50mg，0.116mmol)、5-硼酸嘧啶(28.7mg，0.232mmol)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(16.9mg，0.022mmol)和乙酸钾(34mg，0.35mmol)加入到10/1mL二氧六环/H2O溶液中。在85℃氮气保护下搅拌10小时。将反应液浓缩。以纯EA为展开剂，采用制备TLC纯化得到粗品，用制备HPLC(流动相：ACN/H2O:60/40～20/80)纯化得到目标产物P-26(16.3mg,产率：32.8％)白色固体。Compound 22 (50 mg, 0.116 mmol), 5-boronic acid pyrimidine (28.7 mg, 0.232 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloride palladium (16.9 mg, 0.022 mmol) ) and potassium acetate (34 mg, 0.35 mmol) were added to a 10/1 mL dioxane/H2O solution. Stir under nitrogen protection at 85°C for 10 hours. The reaction solution was concentrated. Using pure EA as the developing solvent, the crude product was purified by preparative TLC, and purified by preparative HPLC (mobile phase: ACN/H2O: 60/40～20/80) to obtain the target product P-26 (16.3 mg, yield: 32.8%) in white solid.
LCMS:t R＝1.181min,MS(ESI)m/z＝430.7[M+H] +. LCMS: t R = 1.181 min, MS (ESI) m/z = 430.7 [M+H] + .
HPLC:t R＝5.285min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =5.285min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(400MHz,DMSO-d6):δ9.23(s,1H),9.04(s,1H),8.71(s,2H),8.39(s,1H),7.67～7.65(m,2H),7.53(s,1H),4.90(t,J＝9.2Hz,1H),4.26(t,J＝8.8Hz,1H),3.69～3.61(m,1H),2.70(d,J＝8.8Hz,3H),1.30(d,J＝6.8Hz,3H). 1 H NMR (400MHz, DMSO-d6): δ9.23(s, 1H), 9.04(s, 1H), 8.71(s, 2H), 8.39(s, 1H), 7.67～7.65(m, 2H), 7.53(s, 1H), 4.90(t, J＝9.2Hz, 1H), 4.26(t, J＝8.8Hz, 1H), 3.69～3.61(m, 1H), 2.70(d, J＝8.8Hz, 3H) ),1.30(d,J=6.8Hz,3H).
实施例28：化合物P-27的制备Example 28: Preparation of Compound P-27
Figure PCTCN2021127847-appb-000083
Figure PCTCN2021127847-appb-000083
将化合物Int-1-1(80mg，0.19mmol)和3,3-二氟环丁烷-1-胺(61.47mg，0.57mmol)溶于DMF(10mL)中。常温下加入HATU(87.29mg，0.23mmol)和DIEA(0.09mL，0.19mmol)。室温下搅拌2h，加入H2O(20mL)并用EA(20mL*2)萃取。有机层用盐水(20ml*3)冲洗，并在真空下浓缩，经TLC(EA)纯化，得到目标中间体23(60mg，产率55.7％)，黄色油状。Compound Int-1-1 (80 mg, 0.19 mmol) and 3,3-difluorocyclobutan-1-amine (61.47 mg, 0.57 mmol) were dissolved in DMF (10 mL). HATU (87.29 mg, 0.23 mmol) and DIEA (0.09 mL, 0.19 mmol) were added at ambient temperature. Stir at room temperature for 2 h, add H2O (20 mL) and extract with EA (20 mL*2). The organic layer was washed with brine (20 ml*3) and concentrated in vacuo, purified by TLC (EA) to give the target intermediate 23 (60 mg, 55.7% yield) as a yellow oil.
LCMS:t R＝1.381min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝506.6,508.6[M+H] +. LCMS: t R = 1.381 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 506.6, 508.6 [M+H] + .
Figure PCTCN2021127847-appb-000084
Figure PCTCN2021127847-appb-000084
将化合物23(50mg,0.12mmol)和嘧啶-5-基硼酸(30.6mg,0.25mmol)溶于二氧六环/H2O(10mL/1mL)中，常温下加入Pd(dppf)Cl2DCM(16.12mg,0.02mmol)和CH 3COOK(30mg,0.3mmol)。88℃搅拌过夜，经硅藻土过滤，滤液真空浓缩。残留物经HPLC(流动相：ACN/H2O＝30/70～80/20)纯化，得到目标产物P-27(15.5mg,产率25.8％)，白色固体。 Compound 23 (50 mg, 0.12 mmol) and pyrimidin-5-ylboronic acid (30.6 mg, 0.25 mmol) were dissolved in dioxane/H2O (10 mL/1 mL), and Pd(dppf)Cl2DCM (16.12 mg, 0.02 mmol) and CH3COOK ( 30 mg, 0.3 mmol). Stir overnight at 88°C, filter through celite, and concentrate the filtrate in vacuo. The residue was purified by HPLC (mobile phase: ACN/H2O=30/70～80/20) to obtain the target product P-27 (15.5 mg, yield 25.8%) as a white solid.
LCMS:t R＝1.298min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝506.7[M+H] +. LCMS: t R = 1.298 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 506.7 [M+H] + .
HPLC:t R＝6.241min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =6.241min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(300MHz,CDCl3):δ9.37(s,1H),8.75(s,2H),8.32(d,J＝9Hz,2H),7.66(d,J＝15Hz,2H),5.63(d,J＝6Hz,1H),4.88(t,J＝9Hz,1H),4.39(brs,1H),4.26(t,J＝9Hz,1H),3.63～3.55(m,1H),3.06～3.00(m,2H),2.52～2.45(m,2H),1.37(d,J＝9Hz,3H). 1 H NMR (300MHz, CDCl3): δ9.37(s, 1H), 8.75(s, 2H), 8.32(d, J=9Hz, 2H), 7.66(d, J=15Hz, 2H), 5.63(d ,J=6Hz,1H),4.88(t,J=9Hz,1H),4.39(brs,1H),4.26(t,J=9Hz,1H),3.63～3.55(m,1H),3.06～3.00( m, 2H), 2.52～2.45 (m, 2H), 1.37 (d, J=9Hz, 3H).
实施例29：化合物P-28的制备Example 29: Preparation of Compound P-28
Figure PCTCN2021127847-appb-000085
Figure PCTCN2021127847-appb-000085
将化合物Int-1-1(60mg,0.144mmol)溶于四氢呋喃(10mL)中，0℃下，加入氯(异丙氧基)甲酮(35.17mg,0.287mmol)和N-甲基吗啡啉(21.77mg,0.215mmol)。反应混合物在0℃下搅拌1h，然后加入环丁基胺(102mg,1.44mmol)，在0℃下继续搅拌1h。反应完，倒入水(20mL)中，乙酸乙酯(2×20mL)萃取。有机层用盐水(20mL)洗涤，无水硫酸镁干燥，过滤，减压浓缩，得到目标中间体24(65mg，产率86.5％)，浅黄色固体。Compound Int-1-1 (60 mg, 0.144 mmol) was dissolved in tetrahydrofuran (10 mL), chloro(isopropoxy) ketone (35.17 mg, 0.287 mmol) and N-methylmorpholine ( 21.77 mg, 0.215 mmol). The reaction mixture was stirred at 0 °C for 1 h, then cyclobutylamine (102 mg, 1.44 mmol) was added and stirring was continued at 0 °C for 1 h. After the reaction was completed, poured into water (20 mL) and extracted with ethyl acetate (2×20 mL). The organic layer was washed with brine (20 mL), dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to give the target intermediate 24 (65 mg, 86.5% yield) as a pale yellow solid.
LCMS:tR＝1.416min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝470.9,472.9[M+H] +. LCMS: tR=1.416min in Shimadzu LC20, chromatography (HALO C18 2.7μm, 4.6mm×50mm), MS(ESI) m/z=470.9, 472.9[M+H] + .
Figure PCTCN2021127847-appb-000086
Figure PCTCN2021127847-appb-000086
将化合物24(65mg,0.138mmol)、嘧啶-5-基硼二醇(25.63mg,0.207mmol)和乙酸钾(27.07mg,0.276mmol)溶于二氧六环/水(11mL,v/v＝10:1)中，加入[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(11.29mg,0.0137mmol)。反应混合物在氮气保护下80℃搅拌10h，然后过滤，减压浓缩。采用flash色谱法(洗脱剂：0％～30％甲醇/二氯甲烷)和prep-HPLC进行分离纯化，得到目标产物合物P-28(31.5mg，产率48.2％)，白色固体。Compound 24 (65 mg, 0.138 mmol), pyrimidin-5-ylborodiol (25.63 mg, 0.207 mmol) and potassium acetate (27.07 mg, 0.276 mmol) were dissolved in dioxane/water (11 mL, v/v= 10:1), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (11.29 mg, 0.0137 mmol) was added. The reaction mixture was stirred at 80°C for 10 h under nitrogen protection, then filtered and concentrated under reduced pressure. Using flash chromatography (eluent: 0%-30% methanol/dichloromethane) and prep-HPLC for separation and purification, the target product compound P-28 (31.5 mg, yield 48.2%) was obtained as a white solid.
LCMS:t R＝1.239min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝470.7[M+H] +. LCMS: t R = 1.239 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 470.7 [M+H] + .
HPLC:t R＝4.429min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =4.429min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(300MHz,DMSO-d 6):δ9.21(s,1H),9.00(s,1H),8.67(s,2H),8.30(s,1H),8.06(d,J＝7.6Hz,1H),7.65(s,1H),7.53(s,1H),4.89(t,J＝9.1Hz,1H),4.36(d,J＝7.8Hz,1H),4.25(t,J＝8.3Hz,1H),3.63(dd,J＝15.3,7.7Hz,1H),2.12(s,2H),1.99(t,J＝9.0Hz,2H),1.61(dd,J＝9.5,5.1Hz,2H),1.29(d,J＝6.8Hz,3H). 1 H NMR (300MHz, DMSO-d 6 ): δ 9.21(s, 1H), 9.00(s, 1H), 8.67(s, 2H), 8.30(s, 1H), 8.06(d, J=7.6Hz ,1H),7.65(s,1H),7.53(s,1H),4.89(t,J＝9.1Hz,1H),4.36(d,J＝7.8Hz,1H),4.25(t,J＝8.3Hz ,1H),3.63(dd,J＝15.3,7.7Hz,1H),2.12(s,2H),1.99(t,J＝9.0Hz,2H),1.61(dd,J＝9.5,5.1Hz,2H) ,1.29(d,J=6.8Hz,3H).
实施例30：化合物P-29的制备Example 30: Preparation of Compound P-29
Figure PCTCN2021127847-appb-000087
Figure PCTCN2021127847-appb-000087
化合物Int-2-1(80mg，0.236mmol)溶于乙酸乙酯(5mL)和四氢呋喃(5mL)中，-60℃下，加入吗啡-4-胺(28.9mg,0.28mmol)，1-丙基磷酸酐(150mg,0.47mmol)和N,N-二异丙基乙胺(121.9mg,0.94mmol)。反应混合物在-60℃搅拌2h，然后减压浓缩。采用prep-HPLC对其进行纯化，得到目标产物P-29(16.9mg，产率16.3％)，白色固体。Compound Int-2-1 (80 mg, 0.236 mmol) was dissolved in ethyl acetate (5 mL) and tetrahydrofuran (5 mL), at -60 °C, morphin-4-amine (28.9 mg, 0.28 mmol), 1-propyl group was added Phosphoric anhydride (150 mg, 0.47 mmol) and N,N-diisopropylethylamine (121.9 mg, 0.94 mmol). The reaction mixture was stirred at -60 °C for 2 h, then concentrated under reduced pressure. It was purified by prep-HPLC to obtain the target product P-29 (16.9 mg, 16.3% yield) as a white solid.
LCMS:t R＝1.175min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝423.7[M+H] +. LCMS: t R = 1.175 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 423.7 [M+H] + .
HPLC:tR＝5.335min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).HPLC:tR=5.335min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(301MHz,DMSO-d6):δ9.43(t,J＝14.6Hz,1H),8.09(s,1H),7.98(s,1H),,7.32(dd,J＝10.5,7.9Hz,1H),6.42(d,J＝10.6Hz,1H),5.73(d,J＝10.6Hz,1H),4.91(t,J＝9.1Hz,1H),4.29(t,J＝8.2Hz,1H),3.74–3.54(m,5H),2.77(dd,J＝10.5,5.8Hz,4H),1.33(dd,J＝6.8,1.6Hz,3H). 1 H NMR (301MHz, DMSO-d6): δ 9.43 (t, J=14.6Hz, 1H), 8.09 (s, 1H), 7.98 (s, 1H), 7.32 (dd, J=10.5, 7.9Hz ,1H),6.42(d,J＝10.6Hz,1H),5.73(d,J＝10.6Hz,1H),4.91(t,J＝9.1Hz,1H),4.29(t,J＝8.2Hz,1H) ), 3.74–3.54 (m, 5H), 2.77 (dd, J=10.5, 5.8Hz, 4H), 1.33 (dd, J=6.8, 1.6Hz, 3H).
实施例31：化合物P-30的制备Example 31: Preparation of Compound P-30
Figure PCTCN2021127847-appb-000088
Figure PCTCN2021127847-appb-000088
在化合物Int-2-1(100mg，0.295mmol)和2-肼基吡啶(38.61mg，0.354mmol)的乙酸乙酯/四氢呋喃＝1：1溶液中-60℃条件下加入1-丙基磷酸酐(140.7mg，0.442mmol)和N，N-二异丙基乙 胺(114.3mg，0.884mmol)。将混合物在-60℃下搅拌1.5小时。旋蒸得到残余物并通过制备型高效液相色谱纯化(流动相：乙腈/水：
Figure PCTCN2021127847-appb-000089
)，得到目标产物P-30(40mg，产率31.5％)，为浅黄色固体。 1-Propylphosphoric anhydride was added to a solution of compound Int-2-1 (100 mg, 0.295 mmol) and 2-hydrazinopyridine (38.61 mg, 0.354 mmol) in ethyl acetate/tetrahydrofuran=1:1 at -60°C (140.7 mg, 0.442 mmol) and N,N-diisopropylethylamine (114.3 mg, 0.884 mmol). The mixture was stirred at -60°C for 1.5 hours. The residue was obtained by rotary evaporation and purified by preparative high performance liquid chromatography (mobile phase: acetonitrile/water:
Figure PCTCN2021127847-appb-000089
) to obtain the target product P-30 (40 mg, yield 31.5%) as a pale yellow solid.
LCMS:t R＝1.137min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝431.1[M+H] +. LCMS: t R = 1.137 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 431.1 [M+H] + .
HPLC:t R＝4.821min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =4.821min in Shimadzu 2010/2030, chromatography (XBRIDGE 2.1×50mm, 3.5um).
1H NMR(400MHz,DMSO):δ10.40(s,1H),9.57(s,1H),8.71(s,1H),8.12(s,1H),8.07(d,J＝3.9Hz,1H),8.01(s,1H),7.54(t,J＝7.2Hz,1H),7.45(d,J＝10.6Hz,1H),6.80-6.65(m,2H),5.98(d,J＝10.6Hz,1H),4.92(t,J＝9.1Hz,1H),4.31(dd,J＝8.7,7.6Hz,1H),3.74-3.64(m,1H),1.34(d,J＝6.9Hz,3H). 1 H NMR (400MHz, DMSO): δ 10.40(s, 1H), 9.57(s, 1H), 8.71(s, 1H), 8.12(s, 1H), 8.07(d, J=3.9Hz, 1H) ,8.01(s,1H),7.54(t,J＝7.2Hz,1H),7.45(d,J＝10.6Hz,1H),6.80-6.65(m,2H),5.98(d,J＝10.6Hz, 1H), 4.92(t, J=9.1Hz, 1H), 4.31(dd, J=8.7, 7.6Hz, 1H), 3.74-3.64(m, 1H), 1.34(d, J=6.9Hz, 3H).
实施例32：化合物P-31的制备Example 32: Preparation of Compound P-31
Figure PCTCN2021127847-appb-000090
Figure PCTCN2021127847-appb-000090
向化合物Int-2-1(50mg，0.147mmol)和1-氨基哌啶-4-醇(20.6mg，0.177mmol)的四氢呋喃溶液(5mL)中添加2-(1H-苯并三唑-1-基)-1,1,3,3-四甲基四氟硼酸盐(56.3mg，0.177mmol)和N、N-二异丙基乙胺(76.2mg，0.59mmol)。将所得混合物在室温下搅拌2.5小时后加入冰水(50mL)并将混合物用乙酸乙酯(3×30mL)萃取。有机相用饱和盐水(50mL)洗涤并用硫酸钠干燥。浓缩得到残留物。通过高效液相色谱法(流动相：乙腈/水:35/65～70/30)纯化，得到目标产物P-31(17mg，产率26.4％)为白色固体。To a solution of compound Int-2-1 (50 mg, 0.147 mmol) and 1-aminopiperidin-4-ol (20.6 mg, 0.177 mmol) in tetrahydrofuran (5 mL) was added 2-(1H-benzotriazole-1- yl)-1,1,3,3-tetramethyltetrafluoroborate (56.3 mg, 0.177 mmol) and N,N-diisopropylethylamine (76.2 mg, 0.59 mmol). The resulting mixture was stirred at room temperature for 2.5 hours after which ice water (50 mL) was added and the mixture was extracted with ethyl acetate (3 x 30 mL). The organic phase was washed with saturated brine (50 mL) and dried over sodium sulfate. Concentration gave a residue. Purification by high performance liquid chromatography (mobile phase: acetonitrile/water: 35/65-70/30) gave the target product P-31 (17 mg, yield 26.4%) as a white solid.
LCMS:tR＝1.240min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝437.7[M+H] +. LCMS: tR=1.240min in Shimadzu LC20, chromatography (HALO C18 2.7μm, 4.6mm×50mm), MS(ESI) m/z=437.7[M+H] + .
1H NMR(301MHz,DMSO):d ppmδ9.19(s,1H),8.94(s,1H),8.13–8.00(m,3H),6.64(d,J＝13.6Hz,1H),4.90(t,J＝9.1Hz,1H),4.62(d,J＝2.8Hz,1H),4.29(t,J＝8.2Hz,1H),3.68(dd,J＝14.5,7.1Hz,1H),3.50(s,1H),2.94–2.81(m,2H),2.55(t,J＝8.8Hz,2H),1.80–1.65(m,2H),1.56–1.41(m,2H),1.37–1.27(m,3H). 1 H NMR (301MHz, DMSO): d ppmδ 9.19(s, 1H), 8.94(s, 1H), 8.13–8.00(m, 3H), 6.64(d, J=13.6Hz, 1H), 4.90(t ,J=9.1Hz,1H),4.62(d,J=2.8Hz,1H),4.29(t,J=8.2Hz,1H),3.68(dd,J=14.5,7.1Hz,1H),3.50(s ,1H),2.94–2.81(m,2H),2.55(t,J=8.8Hz,2H),1.80–1.65(m,2H),1.56–1.41(m,2H),1.37–1.27(m,3H ).
实施例33：化合物P-32的制备Example 33: Preparation of Compound P-32
Figure PCTCN2021127847-appb-000091
Figure PCTCN2021127847-appb-000091
化合物Int-2-1(100mg，0.295mmol)，1-氨基吡咯烷盐酸盐(46.98mg,0.383mmol)和2-(1H-苯并三唑-1-基)-1,1,3,3-四氟硼酸四甲基溴铵(141.98mg,0.44mmol)溶于DMF(5mL)中，氮气保护下，加入N,N-二异丙基乙胺(152mg,1.18mmol)。反应混合物在室温下搅拌16h。然后用水(10mL)稀释，乙酸乙酯(3×10mL)萃取。合并有机层，无水硫酸钠干燥，过滤，减压浓缩。采用prep-HPLC对其进行纯化，得目标产物P-32(31.4mg，产率26.1％)，白色固体。Compound Int-2-1 (100 mg, 0.295 mmol), 1-aminopyrrolidine hydrochloride (46.98 mg, 0.383 mmol) and 2-(1H-benzotriazol-1-yl)-1,1,3, Tetramethylammonium bromide 3-tetrafluoroborate (141.98 mg, 0.44 mmol) was dissolved in DMF (5 mL), and N,N-diisopropylethylamine (152 mg, 1.18 mmol) was added under nitrogen protection. The reaction mixture was stirred at room temperature for 16 h. It was then diluted with water (10 mL) and extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. It was purified by prep-HPLC to obtain the target product P-32 (31.4 mg, 26.1% yield) as a white solid.
LCMS:tR＝1.155min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝407.8[M+H] +. LCMS: tR=1.155min in Shimadzu LC20, chromatography (HALO C18 2.7μm, 4.6mm×50mm), MS(ESI) m/z=407.8[M+H] + .
HPLC:tR＝5.249min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).HPLC:tR=5.249min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(301MHz,DMSO-d6):δ9.24(s,1H),8.99(d,J＝23.7Hz,1H),8.18-8.02(m,3H),,7.25(d,J＝13.9Hz,0.3H),6.65(d,J＝13.6Hz,1H),4.92(t,J＝9.1Hz,1H),4.30(dd,J＝11.7,4.7Hz,1H),3.70(d,J＝7.9Hz,1H),2.81(brs,4H),1.76(d,J＝12.5Hz,4H),1.34(dd,J＝6.8,4.0Hz,3H). 1 H NMR (301MHz, DMSO-d6): δ 9.24 (s, 1H), 8.99 (d, J=23.7Hz, 1H), 8.18-8.02 (m, 3H), 7.25 (d, J=13.9Hz) ,0.3H),6.65(d,J＝13.6Hz,1H),4.92(t,J＝9.1Hz,1H),4.30(dd,J＝11.7,4.7Hz,1H),3.70(d,J＝7.9 Hz, 1H), 2.81 (brs, 4H), 1.76 (d, J=12.5Hz, 4H), 1.34 (dd, J=6.8, 4.0Hz, 3H).
实施例34：化合物P-33的制备Example 34: Preparation of Compound P-33
Figure PCTCN2021127847-appb-000092
Figure PCTCN2021127847-appb-000092
化合物Int-2-1(100mg，0.295mmol)，1-氨基高哌啶(43.76mg,0.383mmol)和2-(1H-苯并三唑-1-基)-1,1,3,3-四氟硼酸四甲基溴铵(141.98mg,0.44mmol)溶于DMF(5mL)中，加入N,N-二异丙基乙胺(152mg,1.18mmol)。反应混合物在室温下搅拌16h。然后用水(10mL)稀释，乙酸乙酯(3×10mL)萃取。合并有机层，无水硫酸钠干燥，过滤，减压浓缩。采用prep-HPLC对其进行纯化，得目标产物 P-33(21.3mg，产率16.2％)，类白色固体。Compound Int-2-1 (100 mg, 0.295 mmol), 1-aminohomopiperidine (43.76 mg, 0.383 mmol) and 2-(1H-benzotriazol-1-yl)-1,1,3,3- Tetramethylammonium bromide tetrafluoroborate (141.98 mg, 0.44 mmol) was dissolved in DMF (5 mL) and N,N-diisopropylethylamine (152 mg, 1.18 mmol) was added. The reaction mixture was stirred at room temperature for 16 h. It was then diluted with water (10 mL) and extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. It was purified by prep-HPLC to obtain the target product P-33 (21.3 mg, yield 16.2%) as an off-white solid.
LCMS:t R＝1.238min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝435.8[M+H] +. LCMS: t R = 1.238 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 435.8 [M+H] + .
HPLC:t R＝6.038min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =6.038min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(301MHz,DMSO-d6):δ9.43(s,1H),8.99(d,J＝16.3Hz,1H),8.16–8.00(m,2.6H),7.38(d,J＝13.9Hz,0.4H),6.67(d,J＝13.6Hz,1H),4.92(t,J＝9.0Hz,1H),4.30(t,J＝8.1Hz,1H),3.70(d,J＝7.4Hz,1H),2.94(brs,4H),1.57(brs,8H),1.34(d,J＝6.7Hz,3H). 1 H NMR (301MHz, DMSO-d6): δ 9.43 (s, 1H), 8.99 (d, J=16.3Hz, 1H), 8.16–8.00 (m, 2.6H), 7.38 (d, J=13.9Hz) ,0.4H),6.67(d,J＝13.6Hz,1H),4.92(t,J＝9.0Hz,1H),4.30(t,J＝8.1Hz,1H),3.70(d,J＝7.4Hz, 1H), 2.94(brs, 4H), 1.57(brs, 8H), 1.34(d, J=6.7Hz, 3H).
实施例35：化合物P-34的制备Example 35: Preparation of Compound P-34
Figure PCTCN2021127847-appb-000093
Figure PCTCN2021127847-appb-000093
化合物Int-2-1(100mg,0.29mmol)和2-肼喹恶啉(56.66mg,0.35mmol)溶于DMF(10mL)中。常温下加入TBTU(113.59mg,0.35mmol)和DIEA(152mg,1.18mmol)。搅拌过夜，加入H2O(20mL)并用EA(20mL*3)萃取。有机层用盐水(20ml*3)冲洗，并在真空下浓缩，经TLC(EA)纯化，得目标产物P-34(4mg，产率2.8％)，黄色固体。Compound Int-2-1 (100 mg, 0.29 mmol) and 2-hydrazinoxaline (56.66 mg, 0.35 mmol) were dissolved in DMF (10 mL). TBTU (113.59 mg, 0.35 mmol) and DIEA (152 mg, 1.18 mmol) were added at room temperature. Stir overnight, add H2O (20 mL) and extract with EA (20 mL*3). The organic layer was washed with brine (20 ml*3), concentrated under vacuum, and purified by TLC (EA) to give the desired product P-34 (4 mg, 2.8% yield) as a yellow solid.
LCMS:t R＝1.236min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝481.7[M+H] +. LCMS: t R = 1.236 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 481.7 [M+H] + .
HPLC:t R＝4.179min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =4.179min in Shimadzu 2010/2030, chromatography (XBRIDGE 2.1×50mm, 3.5um).
1H NMR(300MHz,CD3OD):δ8.82(s,1H),8.45(s,1H),8.25(s,1H),8.20～8.17(m,2H),7.89(d,J＝6Hz,1H),7.75(d,J＝9Hz,1H),7.64(t,J＝6Hz,1H),7.49(t,J＝6Hz,1H),7.02(d,J＝12Hz,1H),4.94(d,J＝9Hz,1H),4.35～4.30(m,1H),3.75～3.65(m,1H),1.43(d,J＝6Hz,3H). 1 H NMR (300MHz, CD3OD): δ8.82(s, 1H), 8.45(s, 1H), 8.25(s, 1H), 8.20～8.17(m, 2H), 7.89(d, J=6Hz, 1H) ), 7.75(d, J＝9Hz, 1H), 7.64(t, J＝6Hz, 1H), 7.49(t, J＝6Hz, 1H), 7.02(d, J＝12Hz, 1H), 4.94(d, J=9Hz,1H),4.35～4.30(m,1H),3.75～3.65(m,1H),1.43(d,J=6Hz,3H).
实施例36：化合物P-35的制备Example 36: Preparation of Compound P-35
Figure PCTCN2021127847-appb-000094
Figure PCTCN2021127847-appb-000094
在化合物Int-2-1(100mg,0.2948mmol)和化合物2(48.88mg,0.3538mmol)的DMF的溶液中(5mL)加入TBTU(113.59mg,0.3538mmol)和DIEA(152.4mg,1.1792mmol).室温搅拌过夜.反应液硅藻土过滤,滤液真空下旋干得到粗产品.经过高效液相色谱得到目标化合物P-35(5.5mg,产率4.07％)，白色固体。To a solution (5 mL) of compound Int-2-1 (100 mg, 0.2948 mmol) and compound 2 (48.88 mg, 0.3538 mmol) in DMF was added TBTU (113.59 mg, 0.3538 mmol) and DIEA (152.4 mg, 1.1792 mmol). The reaction solution was stirred at room temperature overnight. The reaction solution was filtered through celite, and the filtrate was spin-dried under vacuum to obtain the crude product. The target compound P-35 (5.5 mg, yield 4.07%) was obtained through high performance liquid chromatography as a white solid.
LCMS:t R＝1.188min in 10-80AB_7min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*30mm),MS(ESI)m/z＝459.7[M+H] +. LCMS: t R =1.188min in 10-80AB_7min_220&254_Shimadzu.lcm,chromatography(Xtimate C18 2.1*30mm),MS(ESI)m/z=459.7[M+H] + .
HPLC:t R＝5.375min in 10-80AB_15min.met,chromatography(YMC-TriartC18ExRS 150*4.6mml.D S-5um). HPLC: t R =5.375min in 10-80AB_15min.met,chromatography (YMC-TriartC18ExRS 150*4.6mml.D S-5um).
1H NMR(400MHz,DMSO-d 6):δ10.07(s,1H),9.00(s,1H),8.84(s,1H),8.21(s,1H),8.16(d,J＝8Hz,1H),8.06(s,1H),6.85(d,J＝13.7Hz,1H),6.57(s,1H),4.94(t,1H),4.37–4.26(m,1H),3.78–3.65(m,1H),2.24(s,6H),1.36(d,J＝6.9Hz,3H). 1H NMR (400MHz, DMSO-d 6 ): δ 10.07(s, 1H), 9.00(s, 1H), 8.84(s, 1H), 8.21(s, 1H), 8.16(d, J=8Hz, 1H ), 8.06(s, 1H), 6.85(d, J=13.7Hz, 1H), 6.57(s, 1H), 4.94(t, 1H), 4.37–4.26(m, 1H), 3.78–3.65(m, 1H), 2.24(s, 6H), 1.36(d, J=6.9Hz, 3H).
实施例37：化合物P-36的制备Example 37: Preparation of Compound P-36
Figure PCTCN2021127847-appb-000095
Figure PCTCN2021127847-appb-000095
化合物Int-2-1(100mg，0.295mmol)，吡嗪-2-肼(42.2mg,0.383mmol)和2-(1H-苯并三唑-1-基)-1,1,3,3-四氟硼酸四甲基溴铵(141.98mg,0.44mmol)溶于DMF(5mL)中，加入N,N-二异丙基乙胺(152mg,1.18mmol)。反应混合物在室温下搅拌16h。然后用水(10mL)稀释，乙酸乙酯(3×10mL)萃取。合并有机层，无水硫酸钠干燥，过滤，减压浓缩。采用prep-HPLC对其进行纯化，得目标化合物P-36(17.2mg，产率13.5％)，白色固体。Compound Int-2-1 (100 mg, 0.295 mmol), pyrazine-2-hydrazine (42.2 mg, 0.383 mmol) and 2-(1H-benzotriazol-1-yl)-1,1,3,3- Tetramethylammonium bromide tetrafluoroborate (141.98 mg, 0.44 mmol) was dissolved in DMF (5 mL) and N,N-diisopropylethylamine (152 mg, 1.18 mmol) was added. The reaction mixture was stirred at room temperature for 16 h. It was then diluted with water (10 mL) and extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. It was purified by prep-HPLC to obtain the target compound P-36 (17.2 mg, yield 13.5%) as a white solid.
LCMS:t R＝1.203min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS (ESI)m/z＝432.0[M+H] +. LCMS: t R = 1.203 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 432.0 [M+H] + .
HPLC:t R＝6.648min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =6.648min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(301MHz,DMSO-d6):δ10.24(s,1H),9.09(s，1H),8.99(s,1H),,8.46–7.77(m,6H),6.85(d,J＝13.6Hz,1H),4.93(t,J＝9.1Hz,1H),4.36–4.27(m,1H),3.71(dd,J＝15.4,7.5Hz,1H),1.35(d,J＝6.9Hz,3H) 1 H NMR (301MHz, DMSO-d6): δ 10.24(s, 1H), 9.09(s, 1H), 8.99(s, 1H), 8.46–7.77(m, 6H), 6.85(d, J= 13.6Hz, 1H), 4.93 (t, J=9.1Hz, 1H), 4.36–4.27 (m, 1H), 3.71 (dd, J=15.4, 7.5Hz, 1H), 1.35 (d, J=6.9Hz, 3H)
实施例38：化合物P-37的制备Example 38: Preparation of Compound P-37
Figure PCTCN2021127847-appb-000096
Figure PCTCN2021127847-appb-000096
化合物Int-2-1(100mg，0.295mmol)，肼基吡啶(41.82mg,0.383mmol)和2-(1H-苯并三唑-1-基)-1,1,3,3-四氟硼酸四甲基溴铵(141.98mg,0.44mmol)溶于DMF(5mL)中，加入N,N-二异丙基乙胺(152mg,1.18mmol)。反应混合物在室温下搅拌16h。然后用水(10mL)稀释，乙酸乙酯(3×10mL)萃取。合并有机层，无水硫酸钠干燥，过滤，减压浓缩。采用flash色谱法(洗脱剂：0％～30％甲醇/二氯甲烷)和prep-HPLC对其进行纯化，得目标化合物P-37(12mg，产率8.8％)，白色固体。Compound Int-2-1 (100 mg, 0.295 mmol), hydrazinopyridine (41.82 mg, 0.383 mmol) and 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetrafluoroboronic acid Tetramethylammonium bromide (141.98 mg, 0.44 mmol) was dissolved in DMF (5 mL) and N,N-diisopropylethylamine (152 mg, 1.18 mmol) was added. The reaction mixture was stirred at room temperature for 16 h. It was then diluted with water (10 mL) and extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. It was purified by flash chromatography (eluent: 0%-30% methanol/dichloromethane) and prep-HPLC to obtain the target compound P-37 (12 mg, yield 8.8%) as a white solid.
LCMS:tR＝1.040min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝430.8[M+H] +. LCMS: tR=1.040min in Shimadzu LC20, chromatography (HALO C18 2.7μm, 4.6mm×50mm), MS(ESI) m/z=430.8[M+H] + .
HPLC:tR＝4.534min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).HPLC:tR=4.534min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(301MHz,DMSO-d6):δ10.13(s,1H),8.99(s,1H),8.24-7.94(m,6H),,7.16(dd,J＝8.1,4.8Hz,1H),7.05(d,J＝8.2Hz,1H),6.82(d,J＝13.6Hz,1H),4.93(t,J＝9.1Hz,1H),4.36–4.27(m,1H),3.71(dd,J＝15.2,7.3Hz,1H),1.33(d,J＝6.9Hz,3H). 1 H NMR (301MHz, DMSO-d6): δ 10.13 (s, 1H), 8.99 (s, 1H), 8.24-7.94 (m, 6H), 7.16 (dd, J=8.1, 4.8Hz, 1H) ,7.05(d,J＝8.2Hz,1H),6.82(d,J＝13.6Hz,1H),4.93(t,J＝9.1Hz,1H),4.36–4.27(m,1H),3.71(dd, J=15.2, 7.3Hz, 1H), 1.33 (d, J=6.9Hz, 3H).
实施例39：化合物P-38的制备Example 39: Preparation of Compound P-38
Figure PCTCN2021127847-appb-000097
Figure PCTCN2021127847-appb-000097
化合物Int-2-1(100mg，0.295mmol)，4-肼基吡啶盐酸盐(55.8mg,0.383mmol)和2-(1H-苯并三唑-1-基)-1,1,3,3-四氟硼酸四甲基溴铵(141.98mg,0.44mmol)溶于DMF(5mL)中，加入N,N-二异丙基乙胺(152mg,1.18mmol)。反应混合物在室温下搅拌16h。然后用水(10mL)稀释，乙酸乙酯(3×10mL)萃取。合并有机层，无水硫酸钠干燥，过滤，减压浓缩。采用flash色谱法(洗脱剂：0％～30％甲醇/二氯甲烷)和prep-HPLC对其进行纯化，得目标化合物P-38(31.7mg，产率24.8％)，白色固体。Compound Int-2-1 (100 mg, 0.295 mmol), 4-hydrazinopyridine hydrochloride (55.8 mg, 0.383 mmol) and 2-(1H-benzotriazol-1-yl)-1,1,3, Tetramethylammonium bromide 3-tetrafluoroborate (141.98 mg, 0.44 mmol) was dissolved in DMF (5 mL) and N,N-diisopropylethylamine (152 mg, 1.18 mmol) was added. The reaction mixture was stirred at room temperature for 16 h. It was then diluted with water (10 mL) and extracted with ethyl acetate (3 x 10 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. It was purified by flash chromatography (eluent: 0%-30% methanol/dichloromethane) and prep-HPLC to obtain the target compound P-38 (31.7 mg, yield 24.8%) as a white solid.
LCMS:t R＝1.085min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝430.7[M+H] +. LCMS: t R = 1.085 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm×50 mm), MS (ESI) m/z = 430.7 [M+H] + .
HPLC:t R＝5.816min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um). HPLC: t R =5.816min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(300MHz,DMSO-d 6):δ9.07(s,1H),8.52(dd,J＝4.9,1.4Hz,2H),8.26(d,J＝13.8Hz,1H),8.18(s,1H),8.06(s,1H),7.84–7.75(m,3H),5.67(s,2H),4.92(t,J＝9.1Hz,1H),4.32(dd,J＝8.8,7.4Hz,1H),3.70(dd,J＝15.6,7.4Hz,1H),1.35(d,J＝6.9Hz,3H). 1 H NMR (300 MHz, DMSO-d 6 ): δ 9.07 (s, 1H), 8.52 (dd, J=4.9, 1.4 Hz, 2H), 8.26 (d, J=13.8 Hz, 1H), 8.18 (s ,1H),8.06(s,1H),7.84–7.75(m,3H),5.67(s,2H),4.92(t,J=9.1Hz,1H),4.32(dd,J=8.8,7.4Hz, 1H), 3.70(dd, J=15.6, 7.4Hz, 1H), 1.35(d, J=6.9Hz, 3H).
实施例40：化合物P-39的制备Example 40: Preparation of Compound P-39
Figure PCTCN2021127847-appb-000098
Figure PCTCN2021127847-appb-000098
将化合物Int-2-1(80mg,0.29mmol)，环丙基肼(35mg,0.32mmol)，2-(1H-苯并三唑-1-基)-1,1,3,3-四甲基四氟硼酸盐(113mg，0.35mmol)和N，N-二异丙基乙胺(152mg，1.18mmol)混合于N,N-二甲基甲酰胺(6mL)溶液中，将所得混合物在0℃下搅拌2.5小时后向混合物中加入冰水，并用乙酸乙酯萃取(3x30mL)。用饱和食盐水洗涤有机相，并用硫酸钠干燥。浓缩得到残留物。通过高效制备液相色谱(流动相：乙腈/水＝35/65～70/30)纯化，得到目标化合物P-39(10mg,产率8.6％)，为白色固体。Compound Int-2-1 (80 mg, 0.29 mmol), cyclopropylhydrazine (35 mg, 0.32 mmol), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyl tetrafluoroborate (113 mg, 0.35 mmol) and N,N-diisopropylethylamine (152 mg, 1.18 mmol) were mixed in a solution of N,N-dimethylformamide (6 mL), and the resulting mixture was placed in After stirring at 0°C for 2.5 hours, ice water was added to the mixture and extracted with ethyl acetate (3×30 mL). The organic phase was washed with saturated brine and dried over sodium sulfate. Concentration gave a residue. Purification by high performance preparative liquid chromatography (mobile phase: acetonitrile/water=35/65～70/30) gave the target compound P-39 (10 mg, yield 8.6%) as a white solid.
LCMS:t R＝1.297min in Shimadzu LC20,chromatography(HALO C18 2.7μm,4.6mm×50mm),MS(ESI)m/z＝394.2[M+H] +. LCMS: t R = 1.297 min in Shimadzu LC20, chromatography (HALO C18 2.7 μm, 4.6 mm × 50 mm), MS (ESI) m/z = 394.2 [M+H] + .
HPLC:tR＝3.963min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).HPLC:tR=3.963min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(400MHz,DMSO):d ppmδ9.80(d,J＝6.3Hz,1H),8.99(s,1H),8.18–8.12(m,2H),8.04(s, 1H),6.70(d,J＝13.6Hz,1H),5.60(dd,J＝6.1,1.9Hz,1H),4.94(t,J＝9.1Hz,1H),4.32(dd,J＝8.7,7.6Hz,1H),3.75–3.68(m,1H),2.59–2.53(m,1H),1.36(d,J＝6.9Hz,3H),0.45–0.37(m,4H). 1 H NMR (400MHz, DMSO): d ppmδ9.80(d, J=6.3Hz, 1H), 8.99(s, 1H), 8.18-8.12(m, 2H), 8.04(s, 1H), 6.70(d , J=13.6Hz, 1H), 5.60 (dd, J=6.1, 1.9Hz, 1H), 4.94 (t, J=9.1Hz, 1H), 4.32 (dd, J=8.7, 7.6Hz, 1H), 3.75 –3.68 (m, 1H), 2.59–2.53 (m, 1H), 1.36 (d, J=6.9Hz, 3H), 0.45–0.37 (m, 4H).
实施例41：化合物P-40的制备Example 41: Preparation of Compound P-40
Figure PCTCN2021127847-appb-000099
Figure PCTCN2021127847-appb-000099
将化合物25(50mg，0.116mmol，其合成方法与Int-1类似)溶解在10mL THF中。在0度下添加氮甲基吗啡啉(17.6mg，0.17mmol)和氯甲酸异丙酯(28.4mg，0.23mmol)。反应混合液在0℃搅拌30分钟后加入三氟乙胺(115mg，1.16mmol)的四氢呋钠(1mL)溶液，之后继续搅拌1小时。所得到的混合物在真空下浓缩，剩余物用20ml冰水稀释，EA(3x30ml)萃取。合并有机相用是盐水洗，硫酸钠干燥，在真空下浓缩得到粗产品。以石油醚/乙酸乙酯＝1/2为展开剂，通过制备薄层色谱法对其进行纯化，得到化合物26(50mg，产率83.6％)为白色固体。Compound 25 (50 mg, 0.116 mmol, which was synthesized similarly to Int-1) was dissolved in 10 mL of THF. Nitromethylmorpholine (17.6 mg, 0.17 mmol) and isopropyl chloroformate (28.4 mg, 0.23 mmol) were added at 0 degrees. The reaction mixture was stirred at 0°C for 30 minutes, and then a solution of trifluoroethylamine (115 mg, 1.16 mmol) in sodium tetrahydrofuran (1 mL) was added, and stirring was continued for 1 hour. The resulting mixture was concentrated in vacuo, the residue was diluted with 20ml ice water and extracted with EA (3x30ml). The combined organic phases were washed with brine, dried over sodium sulfate, and concentrated in vacuo to give the crude product. Using petroleum ether/ethyl acetate=1/2 as a developing solvent, it was purified by preparative thin layer chromatography to obtain compound 26 (50 mg, yield 83.6%) as a white solid.
LCMS:tR＝1.393min MS(ESI)m/z＝512.6,514.6[M+H]+.LCMS:tR=1.393min MS(ESI)m/z=512.6,514.6[M+H]+.
1H NMR(400MHz,DMSO-d6)δ9.32(s,1H),9.12(t,J＝6.4Hz,1H),8.72(s,1H),8.12(s,1H),8.07(s,1H),4.51(s,2H),4.07～4.00(m,2H),1.40(s,6H).1H NMR(400MHz, DMSO-d6)δ9.32(s,1H),9.12(t,J=6.4Hz,1H),8.72(s,1H),8.12(s,1H),8.07(s,1H) ,4.51(s,2H),4.07～4.00(m,2H),1.40(s,6H).
Figure PCTCN2021127847-appb-000100
Figure PCTCN2021127847-appb-000100
将化合物26(70mg，0.136mmol)、5-硼酸嘧啶(25.3mg，0.2mmol)、[1,1'-双(二苯基膦基)二茂铁]二氯化钯(19.9mg，0.027mmol)和乙酸钾(40mg，0.4mmol)加入到10/1mL二氧六环/H2O溶液中。在90℃氮气保护下搅拌10小时。将反应液浓缩。以纯PE/EA(1/2)为展开剂，采用制备TLC纯化得到粗品，用制备HPLC(流动相：ACN/H2O:55/45～40/60)纯化得到目标产物P-40(19mg, 产率27.0％)白色固体。Compound 26 (70 mg, 0.136 mmol), 5-boronic acid pyrimidine (25.3 mg, 0.2 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloride palladium (19.9 mg, 0.027 mmol) ) and potassium acetate (40 mg, 0.4 mmol) were added to a 10/1 mL dioxane/H2O solution. Stir under nitrogen protection at 90°C for 10 hours. The reaction solution was concentrated. Using pure PE/EA (1/2) as the developing solvent, the crude product was purified by preparative TLC, and purified by preparative HPLC (mobile phase: ACN/H2O: 55/45～40/60) to obtain the target product P-40 (19 mg, yield rate 27.0%) white solid.
LCMS:tR＝1.293min,MS(ESI)m/z＝512.6[M+H] +. LCMS: tR=1.293min, MS(ESI) m/z=512.6[M+H] + .
HPLC:tR＝4.735min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).HPLC:tR=4.735min in Shimadzu 2010/2030,chromatography(XBRIDGE 2.1×50mm,3.5um).
1H NMR(400MHz,DMSO-d6)δ9.26(s,1H),9.07(s,1H),8.73(s,2H),8.45(s,1H),8.41(t,J＝6.4Hz，1H),7.59(s,1H),7.57(s,1H),4.47(s,2H),4.03～3.95(m,2H),1.33(s,6H).1H NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 9.07(s, 1H), 8.73(s, 2H), 8.45(s, 1H), 8.41(t, J=6.4Hz, 1H) ,7.59(s,1H),7.57(s,1H),4.47(s,2H),4.03～3.95(m,2H),1.33(s,6H).
实施例42：化合物对细胞抗增殖实验Example 42: Compounds to Cell Antiproliferative Experiments
准备细胞，96孔板中加入100uL细胞，37℃孵育过夜。第二天DMSO稀释细胞，配制成所需终浓度。3倍稀释样品至0.0015μM，添加50uL样品至细胞培养液中，37℃,5％CO 2孵育72h。平衡至室温后，移取40
Figure PCTCN2021127847-appb-000101
Reagent加入孔板中，混匀，室温孵育60分钟后检测。 To prepare cells, add 100uL of cells to a 96-well plate and incubate at 37°C overnight. The next day cells were diluted in DMSO to make up the desired final concentration. Dilute the sample 3 times to 0.0015μM, add 50uL of the sample to the cell culture medium, and incubate at 37°C, 5% CO 2 for 72h. After equilibrating to room temperature, pipette 40
Figure PCTCN2021127847-appb-000101
Reagent was added to the well plate, mixed well, and incubated at room temperature for 60 minutes before detection.
检测化合物对不同细胞株MM1S(骨髓瘤细胞)、SKM-1(人骨髓增生异常综合征细胞)、LNcap(前列腺癌细胞)、MOLT-4(急性淋巴母细胞白血病细胞)、COLO205(人结肠癌细胞)、RPMI 8226(人多发性骨髓瘤外周血B淋巴细胞)等增殖抑制活性。上述细胞均可通过商业途径购买获得。Test compounds against different cell lines MM1S (myeloma cells), SKM-1 (human myelodysplastic syndrome cells), LNcap (prostate cancer cells), MOLT-4 (acute lymphoblastic leukemia cells), COLO205 (human colon cancer cells) Cells), RPMI 8226 (human multiple myeloma peripheral blood B lymphocytes) and other proliferation inhibitory activities. The above cells can be purchased commercially.
KPT-8602为专利(CN105339358A)中化合物124，结构式如下：KPT-8602 is compound 124 in the patent (CN105339358A), and its structural formula is as follows:
Figure PCTCN2021127847-appb-000102
Figure PCTCN2021127847-appb-000102
【表1】化合物的测定结果(A≤30nM,30nM<B≤100nM,100nM<C≤5μM,D＞5μM,NT:未测试)[Table 1] Measurement results of compounds (A≤30nM, 30nM<B≤100nM, 100nM<C≤5μM, D>5μM, NT: not tested)
表1本发明化合物的细胞抗增殖活性结果Table 1 Results of cell antiproliferative activity of the compounds of the present invention
Figure PCTCN2021127847-appb-000103
Figure PCTCN2021127847-appb-000103
Figure PCTCN2021127847-appb-000104
Figure PCTCN2021127847-appb-000104
表2本发明化合物的细胞抗增殖活性结果Table 2 Results of cell antiproliferative activity of the compounds of the present invention
Figure PCTCN2021127847-appb-000105
Figure PCTCN2021127847-appb-000105
结论：本发明大部分化合物对多种肿瘤细胞的抑制活性明显优于KPT-8602，具有非常强的肿瘤细胞抗增殖活性。Conclusion: Most of the compounds of the present invention have significantly better inhibitory activity on various tumor cells than KPT-8602, and have very strong anti-proliferative activity of tumor cells.
实施例43化合物对核输出蛋白1(XPO1)的抑制活性Inhibitory activity of the compound of Example 43 on nuclear export protein 1 (XPO1)
试验方法：experiment method:
U2OS-Rev(1.4)-NES-mGEP C#52细胞株(绿色荧光蛋白标记HIV Rev响应元件的人骨肉瘤细胞)培养于完全培养基，于37℃,5％CO2至70％～90％融合度。TrypLE消化处理后将细胞重悬于接种培养基中，37℃5％CO2培养过夜。将化合物在DMSO中进行3倍梯度稀释至0.51nM，在用培养基McCoy'5A稀释100倍加入细胞培养板中，孵育4hr。取出细胞培养板，加入100μL 8％PFA(paraformaldehyde solution)，室温固定20min。用PBS洗三遍.加入50μL Hoechst 33342(1:5000),室温避光孵育20min,用PBS再洗三遍,加入50μL PBS于96孔板中，用Operetta读数，选用公式Y＝Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope))，X为对浓度取log值拟合得出IC50值。U2OS-Rev(1.4)-NES-mGEP C#52 cell line (human osteosarcoma cells labeled with HIV Rev response element with green fluorescent protein) were cultured in complete medium at 37℃, 5%CO2 to 70%～90% confluency . After TrypLE digestion, cells were resuspended in inoculation medium and incubated overnight at 37°C with 5% CO2. Compounds were serially diluted 3-fold to 0.51 nM in DMSO, added to cell culture plates at a 100-fold dilution with medium McCoy'5A, and incubated for 4 hr. The cell culture plate was taken out, 100 μL of 8% PFA (paraformaldehyde solution) was added, and it was fixed at room temperature for 20 min. Wash three times with PBS. Add 50 μL of Hoechst 33342 (1:5000), incubate at room temperature for 20 min in the dark, wash three times with PBS, add 50 μL of PBS to a 96-well plate, read with Operetta, select the formula Y=Bottom+(Top- Bottom)/(1+10^((LogIC50-X)*HillSlope)), X is the IC50 value obtained by fitting the log value of the concentration.
表2化合物对核输出蛋白1(XPO1)的抑制活性结果Table 2 Results of inhibitory activity of compounds on nuclear export protein 1 (XPO1)
化合物compound Start Conc.(μM)Start Conc.(μM) IC50(nM)IC50(nM)
P-1P-1 1010 6.0326.032
P-15P-15 1010 5.1665.166
KPT-8602KPT-8602 1010 12.99012.990
结论：本发明化合物P-1和P-15对核输出蛋白1(XPO1)的抑制活性明显优于对照化合物KPT-8602。Conclusion: The inhibitory activity of the compounds P-1 and P-15 of the present invention on nuclear export protein 1 (XPO1) is significantly better than that of the control compound KPT-8602.
实施例44 hERG活性测试Example 44 hERG activity test
试验方法experiment method
使用表达出hERG通道的HEK293细胞，研究本发明化合物对hERG离子通道的影响。化合物均采用利用自身重力的灌流系统进行灌流。每个化合物测试五个浓度，分别为0.3、1、3、10、30μM，每个浓度至少测试两个细胞。在电流稳定(或5分钟)后，再比较化合物使用前后的电流大小变化来计算化合物的阻断作用。将细胞转移到灌流槽中，用细胞外液进行灌流。电极用PC-10(Narishige，Japan)拉制。全细胞膜片钳记录，噪音用采样频率的五分之一进行过滤。将细胞钳制在–80mV，然后用持续4秒方波去极化到40mV，再用持续2秒方波超极化到-40mV，以得到hERG尾电流。这一程序每20秒重复一次。hERG尾电流是纯hERG电流。检测第二个方波引发的最大电流，待其稳定后，灌流测试化合物，当反应稳定后，计算阻断的强度。使用化合物之前的电流数值记为a，灌流化合物至电流稳定时的数值记为b。Using HEK293 cells expressing hERG channels, the effect of compounds of the present invention on hERG ion channels was investigated. Compounds were perfused using a perfusion system utilizing its own gravity. Five concentrations of each compound were tested, 0.3, 1, 3, 10, 30 μM, and at least two cells were tested for each concentration. After the current was stable (or 5 minutes), the current magnitude changes before and after the compound was used were compared to calculate the blocking effect of the compound. The cells were transferred to a perfusion tank and perfused with extracellular fluid. Electrodes were drawn with PC-10 (Narishige, Japan). Whole-cell patch-clamp recordings, with noise filtered at one-fifth of the sampling frequency. Cells were clamped at -80 mV, then depolarized to 40 mV with a 4 s square wave and hyperpolarized to -40 mV with a 2 s square wave to obtain hERG tail currents. This procedure is repeated every 20 seconds. hERG tail currents are pure hERG currents. The maximum current induced by the second square wave was detected. After it stabilized, the test compound was perfused. When the reaction stabilized, the blocking intensity was calculated. The value of the current before the compound was used was denoted as a, and the value when the compound was perfused until the current was stable was denoted as b.
计算方法：阻断率＝(1-a)/bCalculation method: blocking rate = (1-a)/b
平均阻断率为平行检测数据的平均值，将化合物浓度和平均阻断率用origin软件进行拟合，得出IC 50值。 The average blocking rate was the average value of the parallel detection data, and the compound concentration and the average blocking rate were fitted with origin software to obtain the IC 50 value.
表3化合物hERG活性测试结果Table 3 Compound hERG activity test results
化合物compound IC 50(μM) IC50 (μM)
KPT-8602KPT-8602 1.811.81
P-4P-4 2.882.88
p-15p-15 4.724.72
结论：本发明化合物对hERG通道的抑制活性明显低于对照化合物KPT-8602。Conclusion: The inhibitory activity of the compound of the present invention on hERG channel is significantly lower than that of the control compound KPT-8602.
实施例45：本发明化合物的大鼠药代动力学测试Example 45: Rat Pharmacokinetic Testing of Compounds of the Invention
受试物和KPT8602均口服灌胃给药大鼠10mg/kg，给药体积10mL/kg。Both the test substance and KPT8602 were orally administered to rats at 10 mg/kg, and the administration volume was 10 mL/kg.
溶媒：10％DMSO+8％Solutol+15％PEG400in water，配制方法如下：Solvent: 10%DMSO+8%Solutol+15%PEG400in water, the preparation method is as follows:
配制1mg/mL受试物溶液：准确称取10mg受试物，加入1mLDMSO溶液，涡旋超声5min后，加入0.8mL的Solutol溶液，涡旋5分钟后，再加入1.5mL的PEG400溶液，混匀，加水到一定体积10mL即可。Preparation of 1 mg/mL test substance solution: accurately weigh 10 mg of test substance, add 1 mL DMSO solution, vortex for 5 min, add 0.8 mL Solutol solution, vortex for 5 min, add 1.5 mL PEG400 solution, and mix well , add water to a certain volume of 10mL.
于给药后0.25，0.5，1.0，2.0，4.0，6.0，8.0和24h，经颈静脉取血0.2mL，置EDTA-K 2试管中，11000rpm离心5min，分离血浆，LC-MS/MS法检测血浆中原形药物，采用winNonlin计算相关药动学参数T max、C max、AUC 0-t、AUC 0-∞、t 1/2等。研究本发明化合物在大鼠体内的药代动力学行为，评价其药动学特征。 At 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 24 h after administration, 0.2 mL of blood was collected from the jugular vein, placed in an EDTA-K 2 test tube, centrifuged at 11000 rpm for 5 min, the plasma was separated, and detected by LC-MS/MS method For the prototype drug in plasma, winNonlin was used to calculate the relevant pharmacokinetic parameters T max , C max , AUC 0-t , AUC 0-∞ , t 1/2 and so on. The pharmacokinetic behavior of the compound of the present invention in rats was studied, and its pharmacokinetic characteristics were evaluated.
表4化合物的药代动力学参数测定结果The results of determination of pharmacokinetic parameters of the compounds in Table 4
Figure PCTCN2021127847-appb-000106
Figure PCTCN2021127847-appb-000106
结论：p-4、p-10与KPT-8602相比，暴露量明显升高或相当，提示口服吸收良好。P-1，p-15等本发明化合物的口服吸收均较好。CONCLUSION: Compared with KPT-8602, the exposure of p-4 and p-10 was significantly higher or equivalent, suggesting that oral absorption is good. The oral absorption of the compounds of the present invention such as P-1 and p-15 is good.
实施例46：化合物的动物体内药效实验Example 46: In vivo efficacy test of the compound in animals
测试受试样品对人骨髓瘤MM.1S裸鼠异种皮下移植瘤体内生长的抑制作用。The inhibitory effect of the test samples on the growth of human myeloma MM.1S nude mice xenografted subcutaneously in vivo was tested.
(1)细胞培养(1) Cell culture
5％CO 2 37℃培养条件下，MM.1S细胞在含10％胎牛血清RPMI1640培养液中进行常规细胞培养；根据细胞生长情况，每周传代1～2次，传代比例为1：2～3。 MM.1S cells were routinely cultured in RPMI1640 medium containing 10% fetal bovine serum under the culture condition of 5% CO 2 at 37°C; according to cell growth conditions, passage 1 to 2 times a week, and the passage ratio was 1:2～ 3.
(2)建立模型(2) Build a model
用0.25％胰酶消化、吹打细胞使细胞完全分散，收取对数生长期细胞重悬于无血清RPMI1640培养基中，1:1加入Martrigel，调整细胞浓度至6.67*10 7/mL。雌性BALB/c裸小鼠皮下接种人骨髓瘤MM.1S细胞0.15mL，建立MM.1S裸鼠皮下移植瘤模型，定期观察动物及移植瘤生长情况。 Digest the cells with 0.25% trypsin and pipetting to disperse the cells completely, collect the cells in the logarithmic growth phase and resuspend them in serum-free RPMI1640 medium, add Martrigel at 1:1, and adjust the cell concentration to 6.67*10 7 /mL. Female BALB/c nude mice were subcutaneously inoculated with 0.15 mL of human myeloma MM.1S cells to establish the MM.1S nude mice subcutaneously transplanted tumor model, and the growth of the animals and the transplanted tumors were regularly observed.
(3)研究设计(3) Research Design
待平均肿瘤体积生长至100-150mm 3左右时，根据肿瘤体积大小采用随机区组法分组。 When the average tumor volume grew to about 100-150 mm 3 , the patients were randomly divided into groups according to the tumor volume.
口服灌胃给药，对照组采用KPT-8602，给药剂量5mg/kg，实验组给药剂量为2.5、5、7.5、10，每天给药一次，给药周期为4周。Oral administration, the control group was given KPT-8602 at a dose of 5 mg/kg, and the experimental group was administered at a dose of 2.5, 5, 7.5, and 10, once a day, and the administration period was 4 weeks.
给药期间，每周测量2次瘤径，计算肿瘤体积。(肿瘤体积(Tumor volume，TV)计算公式为：TV＝1/2×a×b 2，其中，a表示肿瘤长径；b表示肿瘤短径。) During the administration period, the tumor diameter was measured twice a week, and the tumor volume was calculated. (Tumor volume (TV) is calculated by the formula: TV=1/2×a×b 2 , where a represents the long diameter of the tumor; b represents the short diameter of the tumor.)
试验结束时，剥取皮下肿瘤组织称重，用游标卡尺测量肿瘤直径。肿瘤体积(Tumor volume，TV)计算公式为：TV＝1/2×a×b 2，其中，a表示肿瘤长径；b表示肿瘤短径。 At the end of the experiment, the subcutaneous tumor tissue was excised and weighed, and the diameter of the tumor was measured with a vernier caliper. The calculation formula of tumor volume (TV) is: TV=1/2×a×b 2 , where a represents the long diameter of the tumor; b represents the short diameter of the tumor.
化合物的抑瘤疗效用肿瘤体积抑瘤率(GI)或相对肿瘤增殖率T/C(％)评价。肿瘤体积抑瘤率(GI)计算公式为：GI＝[1－(TV t－TV initial)/(CV t－CV initial)]×100％，其中，TV t表示治疗组每次测量时的瘤体积；TV initial表示分组给药时治疗组的瘤体积；CV t表示对照组每次测量时的瘤体积；CV initial表示分组给药时对照组的瘤体积。相对肿瘤增殖率T/C(％)计算公式为：T/C(％)＝(T RTV/C RTV)×100％，其中，T RTV表示治疗组的相对肿瘤体积，C RTV表示溶剂对照组的相对肿瘤体积。 The tumor inhibitory efficacy of the compounds was evaluated by tumor volume tumor inhibition rate (GI) or relative tumor proliferation rate T/C (%). The tumor volume inhibition rate (GI) was calculated as: GI=[1-(TV t -TV initial )/(CV t -CV initial )]×100%, where TV t represents the tumor in the treatment group at each measurement Volume; TV initial represents the tumor volume of the treatment group when the group is administered; CV t represents the tumor volume of the control group at each measurement; CV initial represents the tumor volume of the control group when the group is administered. The calculation formula of the relative tumor proliferation rate T/C (%) is: T/C (%)=(T RTV /C RTV )×100%, where T RTV represents the relative tumor volume of the treatment group, and C RTV represents the solvent control group relative tumor volume.
统计分析方法：试验数据用Microsoft Office Excel 2010软件进行计算和相关统计学处理。数据用均数±标准误(Mean±S.E)表示。Statistical analysis method: The experimental data were calculated and related statistical processing with Microsoft Office Excel 2010 software. Data are expressed as mean ± standard error (Mean ± S.E).
结论：本发明化合物与对照组相比，具有更强的体内抗肿瘤活性。Conclusion: Compared with the control group, the compounds of the present invention have stronger antitumor activity in vivo.
实施例47：本发明化合物的亚急毒性实验Example 47: Subacute toxicity test of the compounds of the present invention
口服灌胃给予SD大鼠3、9、18mg/kg，给药体积为5mL/kg，，每周给药5天，连续给药2周。Oral gavage was administered to SD rats at 3, 9, and 18 mg/kg, with an administration volume of 5 mL/kg, 5 days a week for 2 consecutive weeks.
溶媒：称取适量Solutol和MC，配制成20％Solutol+0.5％MC，配制方法如下：称取适量受试物样品粉末，研磨，加入适量溶媒，涡旋震荡使样品完全溶解并混匀，超声，配制成所需混悬液，现配现用。Solvent: Weigh an appropriate amount of Solutol and MC to prepare 20% Solutol+0.5% MC. The preparation method is as follows: Weigh an appropriate amount of the sample powder of the test substance, grind, add an appropriate amount of solvent, vortex to completely dissolve the sample and mix well, ultrasonically , prepared into the desired suspension, now with the current use.
检测指标：Detection Indicator:
笼旁观察：给药前及给药后进行笼旁观察，观察内容包括动物外观状况、被毛、体征、行为活动、呼吸状态、腺体分泌、动物姿势、粪便性状、死亡情况等。Observation by the cage: Before and after the administration, the observation by the cage includes the appearance of the animal, coat, physical signs, behavioral activities, respiratory status, gland secretion, animal posture, feces, death, etc.
体重：每周测定1次体重；Body weight: Measure body weight once a week;
临床病理学检查：给药结束测定每组每性别4只大鼠血液学指标、血凝指标、血液生化、电解质指标；Clinicopathological examination: At the end of administration, the hematological indexes, blood coagulation indexes, blood biochemical indexes and electrolyte indexes of 4 rats of each gender in each group were measured;
动物麻醉及解剖：给药结束后对动物实施安乐死，肉眼检查组织器官有无异常，对心脏、肝脏、脾脏、肺脏、肾脏、脑、肾上腺、胸腺、睾丸、附睾、子宫、卵巢。计算脏器系数。Animal anesthesia and dissection: After the administration, the animals were euthanized, and the tissues and organs were visually inspected for abnormalities, such as heart, liver, spleen, lung, kidney, brain, adrenal gland, thymus, testis, epididymis, uterus, and ovary. Calculate the organ coefficient.
公式：脏器系数＝【脏器重量(g)/体重(g)】*100Formula: Organ coefficient = [Organ weight (g) / Body weight (g)] * 100
结论：试验条件下，P-15安全性良好。Conclusion: Under the experimental conditions, P-15 is safe.
以上所述，仅是本发明的较佳实施例而已，并非对本发明作任何形式上的限制，虽然本发明已以较佳实施例揭露如上，然而并非用以限定本发明，任何熟悉本专业的技术人员，在不脱离本发明技术方案范围内，当可利用上述揭示的技术内容作出些许更动或修饰为等同变化的等效实施例，但凡是未脱离本发明技术方案内容，依据本发明的技术实质对以上实施例所作的任何简单修改、等同变化与修饰，均仍属于本发明技术方案的范围内。The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention in any form. Although the present invention has been disclosed above with preferred embodiments, it is not intended to limit the present invention. Technical personnel, within the scope of the technical solution of the present invention, can make some changes or modifications to equivalent examples of equivalent changes by using the technical content disclosed above, provided that the content of the technical solution of the present invention is not deviated from, according to the present invention. Any simple modifications, equivalent changes and modifications made to the above embodiments still fall within the scope of the technical solutions of the present invention.

Claims (23)

  1. 一种化合物，其特征在于，所述化合物为式I所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐；A compound, characterized in that the compound is a compound shown in formula I, or its optical isomer, enantiomer, diastereomer, racemate or racemic mixture, or its solvate , a prodrug, or a pharmaceutically acceptable salt thereof;
    Figure PCTCN2021127847-appb-100001
    Figure PCTCN2021127847-appb-100001
    其中：in:
    R 2、R 3各自独立的选自H或C 1～C 3的烷基； R 2 and R 3 are each independently selected from H or C 1 -C 3 alkyl groups;
    R 4选自H或C 1～C 3的烷基； R 4 is selected from H or C 1 -C 3 alkyl;
    R 1选自未取代或被A任意取代的以下基团：C 6-C 15芳基、5-15元杂芳基、C 6-C 15环烷基或5-15元杂环烷基；取代基A选自：卤素、羟基、羰基(＝O)、-CN、C 1-C 6烷基，C 1-C 6烷氧基、羟基C 1-C 6烷基、卤代C 1-C 6烷基、氰基C 1-C 6烷基、或者未取代或被B任意取代的以下集团：C 6-C 10芳基、5-10元杂芳基、C 6-C 10环烷基、5-10元杂环烷基或
    Figure PCTCN2021127847-appb-100002
    R 5选自H或C 1～C 3的烷基、R 6选自H、C 1-C 6烷基、卤代C 1-C 6烷基、氰基C 1-C 6烷基、C 6-C 10芳基、卤代C 6-C 10芳基、C 1-C 3烷基取代的C 6-C 10芳基；取代基B选自：卤素、羟基、羰基(＝O)、-CN、C 1-C 6烷基、C 1-C 6烷氧基、羟基C 1-C 6烷基、卤代C 1-C 6烷基、氰基C 1-C 6烷基。     R 1 is selected from the following groups unsubstituted or optionally substituted by A: C 6 -C 15 aryl, 5-15 membered heteroaryl, C 6 -C 15 cycloalkyl or 5-15 membered heterocycloalkyl; Substituent A is selected from: halogen, hydroxy, carbonyl (=O), -CN, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, hydroxy C 1 -C 6 alkyl, halogenated C 1 - C 6 alkyl, cyano C 1 -C 6 alkyl, or the following groups unsubstituted or optionally substituted by B: C 6 -C 10 aryl, 5-10 membered heteroaryl, C 6 -C 10 cycloalkane base, 5-10 membered heterocycloalkyl or
    Figure PCTCN2021127847-appb-100002
    R 5 is selected from H or C 1 -C 3 alkyl, R 6 is selected from H, C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, cyano C 1 -C 6 alkyl, C 6 -C 10 aryl, halogenated C 6 -C 10 aryl, C 1 -C 3 alkyl substituted C 6 -C 10 aryl; Substituent B is selected from: halogen, hydroxyl, carbonyl (=O), -CN, C 1 -C 6 alkyl, C 1 -C 6 alkoxy, hydroxy C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, cyano C 1 -C 6 alkyl.
  2. 根据权利要求1所述的化合物，其特征在于，R 2、R 3各自独立的选自H、甲基或乙基。 The compound according to claim 1, wherein R 2 and R 3 are each independently selected from H, methyl or ethyl.
  3. 根据权利要求1所述的化合物，其特征在于，R 4选自H、甲基或乙基。 The compound of claim 1, wherein R 4 is selected from H, methyl or ethyl.
  4. 根据权利要求1所述的化合物，其特征在于，R1选自未取代或任意取代的C 5-C 10芳基；取代基选自卤素、羟基、羰基(＝O)、-CN、C 1-C 3烷基，C 1-C 3烷氧基、羟基C 1-C 3烷基、卤代C 1-C 3烷基、氰基C 1-C 3烷基、C 6-C 10芳基、5-10元杂芳基、C 6-C 10环烷基、5-10元杂环烷。 The compound according to claim 1, wherein R1 is selected from unsubstituted or arbitrarily substituted C 5 -C 10 aryl; substituent is selected from halogen, hydroxyl, carbonyl (=O), -CN, C 1 - C 3 alkyl, C 1 -C 3 alkoxy, hydroxy C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl, cyano C 1 -C 3 alkyl, C 6 -C 10 aryl , 5-10 membered heteroaryl, C 6 -C 10 cycloalkyl, 5-10 membered heterocycloalkane.
  5. 根据权利要求1所述的化合物，其特征在于，R1选自未取代或被A任意取代的5-6元杂芳基；取代基A选自：卤素、羟基、羰基(＝O)、-CN、C 1-C 3烷基，C 1-C 3烷氧基、羟基C 1-C 3烷基、卤代C 1-C 3烷基、氰基C 1-C 3烷基、或者未取代或被B任意取代的以下基团：C 6-C 10芳基、5-10 元杂芳基、C 6-C 10环烷基、5-10元杂环烷基或
    Figure PCTCN2021127847-appb-100003
    R 5选自H或C 1～C 3的烷基、R 6选自H、C 1-C 3烷基、卤代C 1-C 3烷基、氰基C 1-C 3烷基、C 6-C 10芳基、卤代C 6-C 10芳基、C 1-C 3烷基取代的C 6-C 10芳基；取代基B选自：卤素、羟基、羰基(＝O)、-CN、C 1-C 3烷基、C 1-C 3烷氧基、羟基C 1-C 3烷基、卤代C 1-C 3烷基、氰基C 1-C 3烷基。     The compound according to claim 1, wherein R1 is selected from 5-6-membered heteroaryl unsubstituted or arbitrarily substituted by A; Substituent A is selected from: halogen, hydroxyl, carbonyl (=O), -CN , C 1 -C 3 alkyl, C 1 -C 3 alkoxy, hydroxy C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl, cyano C 1 -C 3 alkyl, or unsubstituted Or the following groups optionally substituted by B: C 6 -C 10 aryl, 5-10 membered heteroaryl, C 6 -C 10 cycloalkyl, 5-10 membered heterocycloalkyl or
    Figure PCTCN2021127847-appb-100003
    R 5 is selected from H or C 1 -C 3 alkyl, R 6 is selected from H, C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl, cyano C 1 -C 3 alkyl, C 6 -C 10 aryl, halogenated C 6 -C 10 aryl, C 1 -C 3 alkyl substituted C 6 -C 10 aryl; Substituent B is selected from: halogen, hydroxyl, carbonyl (=O), -CN, C 1 -C 3 alkyl, C 1 -C 3 alkoxy, hydroxy C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl, cyano C 1 -C 3 alkyl.
  6. 根据权利要求1所述的化合物，其特征在于，R1选自未取代或任意取代的8-15元稠杂芳基；取代基选自卤素、羟基、-CN、C 1-C 3烷基，C 1-C 3烷氧基、羟基C 1-C 3烷基、卤代C 1-C 3烷基。 The compound according to claim 1, wherein R1 is selected from unsubstituted or arbitrarily substituted 8-15-membered fused heteroaryl; the substituent is selected from halogen, hydroxyl, -CN, C1 - C3 alkyl, C 1 -C 3 alkoxy, hydroxy C 1 -C 3 alkyl, halogenated C 1 -C 3 alkyl.
  7. 根据权利要求1所述的化合物，其特征在于，R1选自未取代或被任意取代的吡咯基、呋喃基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基、三唑基、噻二唑基或噁二唑基。取代基可以如前任一所述。The compound of claim 1, wherein R1 is selected from unsubstituted or optionally substituted pyrrolyl, furyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazole group, triazolyl, thiadiazolyl or oxadiazolyl. Substituents may be as described above.
  8. 根据权利要求1所述的化合物，其特征在于，R1是具有1、2或3个独立地选自下组的杂原子的任意取代的6元杂芳基，该组由以下各项组成：氮、氧和硫。The compound of claim 1, wherein R1 is an optionally substituted 6-membered heteroaryl group having 1, 2, or 3 heteroatoms independently selected from the group consisting of: nitrogen , oxygen and sulfur.
  9. 根据权利要求1所述的化合物，其特征在于，R1选自未取代或被任意取代的吡啶基、嘧啶基、吡嗪基、哒嗪基或三嗪基。取代基可以如前任一所述。The compound of claim 1, wherein R1 is selected from unsubstituted or optionally substituted pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl or triazinyl. Substituents may be as described above.
  10. 根据权利要求1所述的化合物，其特征在于，所述化合物为式I-1所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐；The compound according to claim 1, wherein the compound is the compound represented by formula I-1, or its optical isomer, enantiomer, diastereomer, racemate or racemate a mixture, or a solvate, prodrug, or a pharmaceutically acceptable salt thereof;
    Figure PCTCN2021127847-appb-100004
    Figure PCTCN2021127847-appb-100004
    其中，R1如权利要求1所述。Wherein, R1 is as described in claim 1.
  11. 根据权利要求1所述的化合物，其特征在于，化合物选自如下：The compound of claim 1, wherein the compound is selected from the group consisting of:
    Figure PCTCN2021127847-appb-100005
    Figure PCTCN2021127847-appb-100005
    Figure PCTCN2021127847-appb-100006
    Figure PCTCN2021127847-appb-100006
    Figure PCTCN2021127847-appb-100007
    Figure PCTCN2021127847-appb-100007
    Figure PCTCN2021127847-appb-100008
    Figure PCTCN2021127847-appb-100008
    Figure PCTCN2021127847-appb-100009
    Figure PCTCN2021127847-appb-100009
    Figure PCTCN2021127847-appb-100010
    Figure PCTCN2021127847-appb-100010
    Figure PCTCN2021127847-appb-100011
    Figure PCTCN2021127847-appb-100011
    Figure PCTCN2021127847-appb-100012
    Figure PCTCN2021127847-appb-100012
    Figure PCTCN2021127847-appb-100013
    Figure PCTCN2021127847-appb-100013
  12. 权利要求1～11任一项所述的化合物的制备方法。The preparation method of the compound of any one of Claims 1-11.
  13. 权利要求10所述的式I-1所示化合物的制备方法：The preparation method of the compound shown in formula I-1 described in claim 10:
    Figure PCTCN2021127847-appb-100014
    Figure PCTCN2021127847-appb-100014
  14. 一种化合物，其特征在于，为式II所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药或其药学上可接受的盐；A compound is characterized in that, it is the compound shown in formula II, or its optical isomer, enantiomer, diastereomer, racemate or racemic mixture, or its solvate, prodrug or a pharmaceutically acceptable salt thereof;
    Figure PCTCN2021127847-appb-100015
    Figure PCTCN2021127847-appb-100015
    其中：环C选自5-6元杂芳基、5-6元杂环基，其中所述杂芳基或杂环基任选进一步被一个或多个R 7取代；优选的，环C选自5-6元杂环基，其中所述杂环基任选进一步被一个或两个R 7取代； Wherein: ring C is selected from 5-6 membered heteroaryl, 5-6 membered heterocyclyl, wherein said heteroaryl or heterocyclyl is optionally further substituted by one or more R 7 ; preferably, ring C is selected from from 5-6 membered heterocyclyl, wherein said heterocyclyl is optionally further substituted with one or two R7 ;
    R 7选自氢原子、卤素、氰基、羟基、C 1～C 8烷基、C 3～C 10环烷基、C 1～C 8烷氧基、C 1～C 8卤代烷基、C 3～C 10卤代环烷基、C 1～C 8卤代烷氧基；优选的，R 7选自氢原子、卤素、氰基、羟基、C 1～C 3烷基、C 3～C 6环烷基、C 1～C 3烷氧基、C 1～C 3卤代烷基、C 3～C 6卤代环烷基、C 1～C 3卤代烷氧基； R 7 is selected from hydrogen atom, halogen, cyano, hydroxyl, C 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, C 1 -C 8 alkoxy, C 1 -C 8 haloalkyl, C 3 ~C 10 halogenated cycloalkyl, C 1 -C 8 halogenated alkoxy; preferably, R 7 is selected from hydrogen atom, halogen, cyano, hydroxyl, C 1 -C 3 alkyl, C 3 -C 6 cycloalkane base, C 1 -C 3 alkoxy, C 1 -C 3 haloalkyl, C 3 -C 6 halocycloalkyl, C 1 -C 3 haloalkoxy;
    R 8选自H、或者未取代或被一个或多个D任意取代的以下基团：C 6～C 10芳基、C 2～C 10杂芳基、C 3～C 10杂环基、C 10～C 20稠环化合物；取代基D选自：氢原子、卤素、羟基、羰基(＝O)、氰基、C 1～C 8烷基，氨基、C 1～C 8烷基取代氨基、羟基取代的C 1～C 8烷基、C 1～C 8卤代烷基、氰基取代的C 1～C 8烷基；优选的，R 8选自H、或者未取代或被一个或多个D任意取代的以下基团：苯、吡咯、呋喃、噻吩、咪唑、噁唑、噻唑、吡唑、嘧啶、喹啉、异喹啉、嘌呤、四唑、三嗪和咔唑或萘；取代基D选自：氢原子、卤素、羟基、羰基(＝O)、氰基、C 1～C 3烷基，氨基、C 1～C 3烷基取代氨基、羟基取代的C 1～C 3烷基、C 1～C 3卤代烷基、氰基取代的C 1～C 3烷基； R 8 is selected from H, or the following groups unsubstituted or optionally substituted by one or more D: C 6 -C 10 aryl, C 2 -C 10 heteroaryl, C 3 -C 10 heterocyclyl, C 10 - C20 fused ring compound; Substituent D is selected from: hydrogen atom, halogen, hydroxyl, carbonyl (=O), cyano, C1 - C8 alkyl, amino, C1 - C8 alkyl substituted amino, Hydroxy-substituted C 1 -C 8 alkyl, C 1 -C 8 haloalkyl, cyano-substituted C 1 -C 8 alkyl; preferably, R 8 is selected from H, or unsubstituted or by one or more D Optionally substituted: benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyrimidine, quinoline, isoquinoline, purine, tetrazole, triazine and carbazole or naphthalene; Substituent D Selected from: hydrogen atom, halogen, hydroxyl, carbonyl (=O), cyano group, C 1 -C 3 alkyl, amino, C 1 -C 3 alkyl substituted amino, hydroxyl substituted C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, cyano-substituted C 1 -C 3 alkyl;
    R 9选自未取代或被一个或多个E任意取代的以下基团：羟基、氨基；取代基E选自：卤素、羟基、C 1～C 6烷基，C 3～C 10环烷基、C 2～C 10杂环基、C 1～C 6烷氧基、C 1～C 6卤代烷基、C 3～C 10卤代环烷基、
    Figure PCTCN2021127847-appb-100016
    或与被取代的原子形成4～8元杂环；R 9选自：氢原子、羟基、C 1～C 6烷基，C 3～C 10环烷基、C 1～C 6烷氧基、C 1～C 6卤代烷基、C 3～C 10卤代环烷基、C 6～C 10芳基、C 2～C 10杂芳基或C 2～C 10杂环基。     R 9 is selected from the following groups unsubstituted or optionally substituted by one or more E: hydroxyl, amino; Substituent E is selected from: halogen, hydroxyl, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl , C 2 -C 10 heterocyclyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 3 -C 10 halocycloalkyl,
    Figure PCTCN2021127847-appb-100016
    Or form a 4-8 membered heterocycle with the substituted atom; R 9 is selected from: hydrogen atom, hydroxyl, C 1 -C 6 alkyl, C 3 -C 10 cycloalkyl, C 1 -C 6 alkoxy, C 1 -C 6 haloalkyl, C 3 -C 10 halocycloalkyl, C 6 -C 10 aryl, C 2 -C 10 heteroaryl or C 2 -C 10 heterocyclic.
  15. 根据权利要求14所述的化合物，其特征在于，R 9选自未取代或被一个或多个E任意取代的以下基团：羟基、氨基；取代基E选自：卤素、羟基、C 1～C 4烷基，C 3～C 8环烷基、C 2～C 8杂环基、C 1～C 4烷氧基、C 1～C 4卤代烷基、C 3～C 8卤代环烷基、
    Figure PCTCN2021127847-appb-100017
    或与被取代的原子形成4～6元杂环；R 9选自：氢原子、羟基、C 1～C 3烷基，C 3～C 8环烷基、C 1～C 3烷氧基、C 1～C 3卤代烷基、C 3～C 8卤代环烷基、C 6～C 10芳基、C 2～C 8杂芳基或C 2～C 8杂环基。     The compound according to claim 14, wherein R 9 is selected from the following groups unsubstituted or optionally substituted by one or more E: hydroxyl, amino; Substituent E is selected from: halogen, hydroxyl, C 1 ~ C 4 alkyl, C 3 -C 8 cycloalkyl, C 2 -C 8 heterocyclyl, C 1 -C 4 alkoxy, C 1 -C 4 haloalkyl, C 3 -C 8 halocycloalkyl ,
    Figure PCTCN2021127847-appb-100017
    Or form a 4-6 membered heterocycle with the substituted atom; R 9 is selected from: hydrogen atom, hydroxyl, C 1 -C 3 alkyl, C 3 -C 8 cycloalkyl, C 1 -C 3 alkoxy, C 1 -C 3 haloalkyl, C 3 -C 8 halocycloalkyl, C 6 -C 10 aryl, C 2 -C 8 heteroaryl or C 2 -C 8 heterocyclic.
  16. 根据权利要求14所述的化合物，其特征在于，所述化合物为如式II-1或式II-2所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药或其药学上可接受的盐：The compound according to claim 14, wherein the compound is a compound represented by formula II-1 or formula II-2, or its optical isomer, enantiomer, diastereomer, racem A isomer or racemic mixture, or a solvate, prodrug or a pharmaceutically acceptable salt thereof:
    Figure PCTCN2021127847-appb-100018
    Figure PCTCN2021127847-appb-100018
    其中，R 8和R 9如权利要求1所述；优选的，R 8为H或嘧啶。 Wherein, R 8 and R 9 are as described in claim 1; preferably, R 8 is H or pyrimidine.
  17. 如下所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药或其药学上可接受的盐：The compounds shown below, or their optical isomers, enantiomers, diastereomers, racemates or racemic mixtures, or solvates, prodrugs or pharmaceutically acceptable salts thereof:
    Figure PCTCN2021127847-appb-100019
    Figure PCTCN2021127847-appb-100019
    Figure PCTCN2021127847-appb-100020
    Figure PCTCN2021127847-appb-100020
    Figure PCTCN2021127847-appb-100021
    Figure PCTCN2021127847-appb-100021
    Figure PCTCN2021127847-appb-100022
    Figure PCTCN2021127847-appb-100022
    Figure PCTCN2021127847-appb-100023
    Figure PCTCN2021127847-appb-100023
  18. 权利要求14～17任一项所述的化合物的制备方法。The preparation method of the compound of any one of Claims 14-17.
  19. 一种药物组合物，其包括权利要求1～11，14～17任一项所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐，以及药学上可接受的载体。A pharmaceutical composition comprising the compound shown in any one of claims 1 to 11, 14 to 17, or an optical isomer, enantiomer, diastereomer, racemate or racemic mixture thereof , or a solvate, prodrug thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  20. 权利要求1～11，14～17任一项所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐或权利要求14所述的组合物作为XPO1抑制剂的应用。The compound shown in any one of claims 1 to 11, 14 to 17, or an optical isomer, enantiomer, diastereomer, racemate or racemic mixture thereof, or a solvate, pro- Use of the medicine, or a pharmaceutically acceptable salt thereof or the composition of claim 14 as an XPO1 inhibitor.
  21. 权利要求1～11，14～17任一项所示的化合物，或其旋光异构体、对映体、非对映体、外消旋体或外消旋混合物，或其溶剂合物、前药，或其药学上可接受的盐或权利要求14所述的组合物作为制备治疗多种与不当的核转运触发的异常细胞应答相关的疾病、紊乱或病症的药物中的用途。The compound shown in any one of claims 1 to 11, 14 to 17, or an optical isomer, enantiomer, diastereomer, racemate or racemic mixture thereof, or a solvate, pro- Use of a medicament, or a pharmaceutically acceptable salt thereof, or the composition of claim 14 for the manufacture of a medicament for the treatment of various diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport.
  22. 根据权利要求21所述的用途，其特征在于，所述的多种与不当的核转运触发的异常细胞应答相关的疾病、紊乱或病症包括但不限于增生性紊乱、癌症、炎症性紊乱、自身免疫性紊乱、病毒感染、神经退行性紊乱、眼科紊乱、异常组织生长紊乱、与食物摄取有关的紊乱、过敏以及呼吸紊乱。The use according to claim 21, wherein the various diseases, disorders or conditions associated with abnormal cellular responses triggered by inappropriate nuclear transport include, but are not limited to, proliferative disorders, cancer, inflammatory disorders, autologous Immune disorders, viral infections, neurodegenerative disorders, ophthalmic disorders, abnormal tissue growth disorders, disorders related to food intake, allergies, and respiratory disorders.
  23. 根据权利要求21所述的用途，其特征在于，所述癌症为淋巴瘤、脂肪肉瘤、多发性骨髓瘤、骨髓增生异常综合症、前列腺癌、结直肠癌、子宫内膜癌、胰腺癌、胃癌、弥漫性大B细胞淋巴瘤、非小细胞肺癌、卵巢癌、乳腺癌、急性髓细胞性白血病、胸腺瘤、食道癌、胶质母细胞瘤和其它实体瘤。The use according to claim 21, wherein the cancer is lymphoma, liposarcoma, multiple myeloma, myelodysplastic syndrome, prostate cancer, colorectal cancer, endometrial cancer, pancreatic cancer, gastric cancer , diffuse large B-cell lymphoma, non-small cell lung cancer, ovarian cancer, breast cancer, acute myeloid leukemia, thymoma, esophageal cancer, glioblastoma and other solid tumors.
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