WO2020259205A1 - DERIVATIVES OF OLEANOLIC ACID AND δ-OLEANOLIC ACID AND MEDICAL USE THEREOF - Google Patents

DERIVATIVES OF OLEANOLIC ACID AND δ-OLEANOLIC ACID AND MEDICAL USE THEREOF Download PDF

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WO2020259205A1
WO2020259205A1 PCT/CN2020/093248 CN2020093248W WO2020259205A1 WO 2020259205 A1 WO2020259205 A1 WO 2020259205A1 CN 2020093248 W CN2020093248 W CN 2020093248W WO 2020259205 A1 WO2020259205 A1 WO 2020259205A1
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compound
300mhz
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孙宏斌
胡凯文
刘柳
程亚龙
秦鹿柘
李浩斌
戴量
柳军
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中国药科大学
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Definitions

  • the present invention relates to the field of biomedicine, relates to novel pentacyclic triterpenoids with AMPK agonistic activity, and specifically relates to derivatives of oleanolic acid and delta-oleanolic acid and their medical uses.
  • the present invention also relates to such compounds in Use in preparing medicines for preventing or treating AMPK-mediated diseases and their pharmaceutical compositions.
  • AMPK Adrenylate Activated Protein Kinase
  • ERR ⁇ estrogen-related receptor ⁇
  • AMPK-mediated diseases include metabolic diseases and cardiovascular and cerebrovascular diseases, such as insulin resistance, metabolic syndrome, type 1 or type 2 diabetes, hyperlipidemia, obesity, atherosclerosis, myocardial ischemia, myocardial infarction, Arrhythmia, coronary heart disease, hypertension, heart failure, myocardial hypertrophy, myocarditis, diabetic complications (including diabetic cardiomyopathy, diabetic nephropathy, retinopathy, neuropathy and diabetic ulcers, etc.), non-alcoholic fatty liver, non-alcoholic fat Hepatitis, alcoholic fatty liver, cirrhosis, gout, stroke or cerebral infarction, etc.; AMPK-mediated diseases also include inflammatory diseases, autoimmune diseases, organ fibrosis diseases, neurological damage diseases, or subsequent infections caused by pathogens Primary diseases, such as pneumonia, asthma, chronic obstructive pulmonary disease, chronic bronchitis, emphylococcus, etc.
  • AMPK agonists can prevent and treat a variety of AMPK-mediated diseases (J.Med.Chem.,2015,58 2; Nature 2013,493,346; Experimental Neurology 2017,298,31; Biochemical Pharmacology 2010,80,1708; Current Drug Targets, 2016, 17, 908; Nat Rev Drug Discov, 2019, DOI: 10.1038/s41573-019-0019-2).
  • metformin a hypoglycemic drug widely used clinically, is believed to exert a variety of clinical effects by activating AMPK (J. Clin. Invest. 2001, 108, 1167).
  • AMPK agonists have broad clinical application prospects, so far, no substantial progress has been made in the development of new safe and effective AMPK agonists.
  • AMPK- ⁇ subunit agonist MK-8722 can lower blood sugar
  • AICAR as one of the most commonly used AMPK agonists (Eur. J. Biochem. 1995, 229, 558), has also been terminated in clinical trials due to its relatively large side effects (J Clin Pharmacol 1991, 31: 342-347) .
  • oleanolic acid is a common pentacyclic triterpene in medicinal plants, which has a wide range of biological activities (Nat Prod Rep 2011, 28, 543).
  • ⁇ -oleanolic acid 3 ⁇ -hydroxy-oleanan-13(18)-en-28-acid
  • ⁇ -oleanolic acid is a pentacyclic triterpene acid that is very rare in nature (Phytochemistry 1999, 51, 83). So far, there are very few reports on the biological activity of ⁇ -oleanolic acid.
  • the present invention provides a novel oleanolic acid and ⁇ -oleanolic acid derivatives; the new oleanolic acid derivative or ⁇ -Oleanolic acid derivatives are a new type of AMPK agonist, so they can be used to prepare drugs for preventing or treating AMPK-mediated diseases.
  • the inventors unexpectedly discovered a series of potent new AMPK agonists when they modified the structure of ⁇ -oleanolic acid and oleanolic acid, and their AMPK agonist activity was significantly better than the recognized AMPK agonist AICAR .
  • R is H, Or R a CO-;
  • R 1 is H, C 1 -C 5 alkyl substituted by substituent Y or a C 1 -C 5 alkyl
  • the substituent Y is OH, C (O) OH, C (O) NH 2, NH 2, NHC(O)CH 3 , pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorph Lin-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethylammonium or diethanolamino;
  • R a is an unsubstituted or substituted group L is substituted C 1 -C 5 alkyl, the substituent group L is one or two substituents independently selected from the group: OH, C (O) OH , NH 2, pyrrolidin Alk-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorpholine-1,1-diox Generation-4-yl or NHC(O)CH(CH 3 )NHC(O)CH(CH 3 )NH;
  • R' is OR 2 , NR 3 R 4 or
  • R 2 is a C 1 -C 5 alkyl group substituted by a substituent Z, and the substituent Z is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium or diethanolamine group;
  • R 3 is H or C 1 -C 3 alkyl
  • R 4 is a C 1 -C 5 alkyl group substituted by a substituent W, the substituent W is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium, diethanolamine or acetylamino;
  • R 5 is H, OH, F, NH 2 , C 1 -C 3 alkylamino, COOH, C 1 -C 3 alkyl, C 1 -C 3 alkoxy, cycloalkane attached to any carbon on the ring Oxy or heterocycloalkyloxy;
  • Q is CH 2 , O, NR 6 , S, SO 2 , CHR 7 or chemical bond;
  • R 6 is H, C 1 -C 3 alkyl, C 1 -C 3 alkyl acyl, C 1 -C 3 alkylsulfonyl, or C 1 -C 3 alkyl substituted by the substituent P, the substituent P Is OH, COOH, NH 2 or C 1 -C 3 alkylamino;
  • R 7 is OH, NH 2 , COOH, C 1 -C 3 alkylamino, C 1 -C 3 alkoxy, cycloalkyloxy or heterocycloalkyloxy;
  • n 0, 1 or 2;
  • n 0, 1, or 2.
  • the compound of Formula I or Formula II, or a pharmaceutically acceptable salt or ester or solvate thereof is a pharmaceutically acceptable salt or ester or solvate thereof.
  • R is H, Or R a CO-;
  • R 1 is H, C 1 -C 5 alkyl substituted by substituent Y or a C 1 -C 5 alkyl
  • the substituent Y is OH, C (O) OH, C (O) NH 2, NH 2, NHC(O)CH 3 , pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorph Lin-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethylammonium or diethanolamino;
  • R a is an unsubstituted or substituted group L is substituted C 1 -C 5 alkyl, the substituent group L is one or two substituents independently selected from the group: OH, C (O) OH , NH 2, pyrrolidin Alk-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorpholine-1,1-diox Generation-4-yl or NHC(O)CH(CH 3 )NHC(O)CH(CH 3 )NH;
  • R' is OR 2 , NR 3 R 4 or
  • R 2 is a C 1 -C 5 alkyl group substituted by a substituent Z, and the substituent Z is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium or diethanolamine;
  • R 3 is H;
  • R 4 is a C 1 -C 5 alkyl group substituted by a substituent W, the substituent W is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium, diethanolamine or acetylamino;
  • R 5 is H, OH, F, NH 2 , C 1 -C 3 alkylamino or COOH attached to any carbon on the ring;
  • Q is CH 2 , O, NR 6 , S, SO 2 or CHR 7 ;
  • R 6 is H, C 1 -C 3 alkyl, C 1 -C 3 alkyl acyl, C 1 -C 3 alkylsulfonyl or C 1 -C 3 alkyl substituted with substituent P, said substituent P Is OH, COOH, NH 2 or C 1 -C 3 alkylamino;
  • R 7 is OH, NH 2 , COOH or C 1 -C 3 alkylamino
  • n 0 or 1
  • m 0 or 1.
  • the compound or a pharmaceutically acceptable salt or ester or solvate thereof is selected from the following compounds:
  • the compounds of the present invention can also be used as pharmaceutical salts.
  • the salt may be the acid salt of at least one of the following acids: galactonic acid, D-glucuronic acid, glycerophosphoric acid, hippuric acid, isethionic acid, lactobionic acid, maleic acid, 1,5-naphthalene Disulfonic acid, naphthalene-2-sulfonic acid, pivalic acid, terephthalic acid, thiocyanic acid, cholic acid, n-dodecyl sulfuric acid, benzenesulfonic acid, citric acid, D-glucose, glycolic acid, lactic acid, Malic acid, malonic acid, mandelic acid, phosphoric acid, propionic acid, hydrochloric acid, sulfuric acid, tartaric acid, succinic acid, formic acid, hydroiodic acid, hydrobromic acid, methanesulfonic acid, niacin, nitric acid, orotic acid, ox
  • the salt can also be the compound of the present invention and metal (including sodium, potassium, calcium, etc.) ions or pharmaceutically acceptable amines (including ethylenediamine, tromethamine, etc.), ammonium ions or choline
  • metal including sodium, potassium, calcium, etc.
  • pharmaceutically acceptable amines including ethylenediamine, tromethamine, etc.
  • ammonium ions or choline The salt formed.
  • the compounds of the present invention can also be formed into pharmaceutical compositions in the form of esters, prodrugs, N-oxides or their solvates.
  • Oleanolic acid can be purchased commercially.
  • the synthesis of ⁇ -oleanolic acid can be carried out with reference to the literature method (Org.Biomol.Chem., 2016, 14, 11154), that is, the benzyl ester of ⁇ -oleanolic acid is prepared by referring to the literature method, and then subjected to conventional catalysis Hydrogenation debenzylation can produce ⁇ -oleanolic acid.
  • the synthesis of ⁇ -oleanolic acid and oleanolic acid derivatives can be carried out by referring to the method in the examples or the improved method.
  • the present invention provides the use of the compound of formula I or formula II, its pharmaceutically acceptable salt or ester or solvate in the preparation of AMPK agonists with the activity of enhancing AMPK phosphorylation level.
  • the compound of the present invention has significant agonistic activity on AMPK, and therefore can be used to prepare AMPK agonists with activity to enhance AMPK phosphorylation level.
  • the present invention also provides the use of the compound of formula I or formula II, its pharmaceutically acceptable salt or ester or solvate in the preparation of a medicine for preventing or treating AMPK-mediated diseases.
  • the AMPK-mediated diseases include metabolic diseases, cardiovascular and cerebrovascular diseases, inflammatory diseases, autoimmune diseases, organ fibrotic diseases, neurodegenerative diseases, secondary diseases caused by pathogen infection, and mitochondrial dysfunction Or disorder disease or tumor.
  • the AMPK-mediated diseases include: insulin resistance, metabolic syndrome, type 1 or type 2 diabetes, hyperlipidemia, obesity, atherosclerosis, myocardial deficiency Blood, myocardial infarction, arrhythmia, coronary heart disease, hypertension, heart failure, myocardial hypertrophy, myocarditis, diabetic complications (including diabetic cardiomyopathy, diabetic nephropathy, retinopathy, neuropathy, diabetic ulcers, etc.), non-alcoholic fatty liver , Non-alcoholic steatohepatitis, alcoholic fatty liver, cirrhosis, gout, stroke or cerebral infarction, etc.
  • the AMPK-mediated diseases such as inflammatory diseases, autoimmune diseases, organ fibrotic diseases, neurological damage diseases, or secondary diseases caused by pathogen infection, include: pneumonia, asthma, chronic obstructive pulmonary disease, chronic bronchus Inflammation, emphysema, bronchiolitis obliterans, idiopathic pulmonary fibrosis, cystic fibrotic lung disease, allergic rhinitis, inflammatory bowel disease (such as Crohn’s disease and ulcerative colitis), polycystic kidney disease, Polycystic ovary syndrome (PCOS), Behcet's disease, systemic lupus erythematosus, rheumatoid arthritis, spondyloarthritis, osteoarthritis, synovitis, tendinitis, thromboangiitis obliterans, phlebitis, intermittent Claudication, keloid, psoriasis, ichthyosis, bullous pemphi
  • the AMPK-mediated diseases such as mitochondrial dysfunction and disorder, include: myasthenia, myoclonus, exercise intolerance, Karnes-Saier syndrome, chronic fatigue syndrome, Leigh syndrome, mitochondria Myopathy-encephalopathy-hyperlactic acidemia, stroke syndrome or stroke-like episodes.
  • the compounds of the present invention can also be used to treat muscular dystrophy conditions, for example, Duchenne muscular dystrophy, conch muscular dystrophy, or Friedrich's ataxia.
  • the AMPK-mediated diseases include: bone cancer, acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, and myeloproliferation Abnormal syndrome, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hemangioma, granuloma, xanthoma, meningiosarcoma, glioma, astrocytoma, medulloblastoma, ependymal Tumor, germ cell tumor (pineal tumor), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, fibroneuronoma, sarcoma, esophageal cancer, gastric cancer, pancreas Cancer, colorectal cancer, colon cancer
  • the pharmaceutical composition for preventing or treating AMPK-mediated diseases of the present invention contains a therapeutically effective amount of a compound of formula I or formula II or a pharmaceutically acceptable salt or ester or solvate thereof as an active ingredient and a pharmaceutical composition Acceptable excipients.
  • the adjuvants that can be mixed arbitrarily can be changed according to the dosage form and administration form. Examples of excipients include excipients, binders, disintegrants, lubricants, correctives, flavors, coloring agents, or sweeteners.
  • the pharmaceutical composition can be in the form of capsules, powders, tablets, granules, pills, injections, syrups, oral liquids, inhalants, ointments, suppositories or patches, etc.
  • the compound of formula I or formula II of the present invention or a pharmaceutically acceptable salt or ester or solvate thereof can be used in combination with one or more other types of drugs for preventing or treating AMPK-mediated diseases, including but It is not limited to the following combinations of drugs.
  • the drugs that can be used in combination with the compounds of the present invention can be one or more anti-diabetic drugs, including metformin, sulfonylurea hypoglycemic drugs (such as glibenclamide and glimepiride, etc.), glucosidase inhibitors (Such as acarbose and miglitol), PPAR ⁇ agonists (such as pioglitazone and rosiglitazone), PPAR ⁇ / ⁇ dual agonists, dipeptidyl peptidase IV (DPP-IV) inhibitors (such as Sitagliptin, saxagliptin, alogliptin and linagliptin, etc.), Glinide-type hypoglycemic drugs (such as repaglinide and nateglinide, etc.), SGLT2 inhibitors (such as Kangel Liegliflozin, Dapagliflozin, Enpagliflozin, Igliglifl
  • the drugs that can be used in combination with the compounds of the present invention can be one or more weight loss drugs, including lorcaserin, orlistat, and glucagon-like peptide-1 (GLP-1) drugs (such as A Sernatide, liraglutide, lisnatide, duraglutide, benaglutide and abiglutide, etc.).
  • GLP-1 glucagon-like peptide-1
  • the drugs that can be used in combination with the compounds of the present invention can be one or more anti-non-alcoholic fatty liver disease drugs, including: AMPK agonists (such as metformin), farnesoid X receptor (FXR) agonists (such as Obeticholic acid, GS-9674, EDP-305 and LJN452, etc.), acetyl-Coenzyme A carboxylase (ACC) inhibitors (such as GS-0976, etc.), apoptosis signal-regulated kinase-1 (ASK1) inhibitors (such as ceremoniessertib, etc.), PPAR agonists (such as Elafibranor, Saroglitazar, IVA337 and MSDC-0602K, etc.), caspase inhibitors (such as Emricasan, etc.), stearoyl-CoA desaturase 1 (SCD1) inhibition Agents (such as Aramchol, etc.), long-acting glucagon-like peptide
  • the drugs that can be used in combination with the compounds of the present invention can be one or more lipid-lowering drugs, including niacin, statins (such as lovastatin, simvastatin, pravastatin, mevastatin, fluvastatin) Statins, atorvastatin, cerivastatin, rovastatin and pitavastatin), cholesterol absorption inhibitors (such as ezetimibe, etc.), fibrates (such as clofibrate, bezafibrate, fenofibrate) Special class), PCSK9 inhibitors (such as Evolocumab and Alirocumab, etc.), CETP inhibitors (such as anacetrapib, etc.), AMPK agonists and ACC inhibitors (such as GS-0976, etc.).
  • statins such as lovastatin, simvastatin, pravastatin, mevastatin, fluvastatin
  • Statins such as lovastatin, simva
  • the dosage of the compound of formula I or formula II of the present invention or a pharmaceutically acceptable salt or ester or solvate thereof can be appropriately changed according to the patient's age, weight, symptoms and administration route.
  • the dosage of the compound of formula I or formula II or a pharmaceutically acceptable salt or ester or solvate thereof is 1 mg to 1000 mg/time, preferably 5 mg to 500 mg/time, more preferably 10mg ⁇ 60mg/time, 1 to 3 times a day. It can also deviate from this dosage range according to the degree of disease and the different dosage forms.
  • the present invention has the following advantages:
  • novel derivatives of oleanolic acid and delta-oleanolic acid of the present invention have potent AMPK agonist activity, and their activity is significantly better than the recognized AMPK agonist AICAR.
  • the novel pentacyclic triterpenoids of the present invention have better oral bioavailability and other pharmacokinetic properties.
  • the novel pentacyclic triterpenoids of the present invention have very Good security.
  • the novel pentacyclic triterpenoids of the present invention have the advantages of low cost, easy preparation and less potential side effects than existing AMPK agonists. They can be used alone or combined with one Or a variety of other types of drugs for preventing or treating AMPK-mediated diseases are used in combination, which is expected to become a new type of drug for preventing or treating AMPK-mediated diseases.
  • Figure 1 shows the effect of some compounds on the downstream signal pathway of AMPK in Huh-7 cells (Western Blot detection).
  • N,N-Dimethylethylenediamine (66 ⁇ L, 0.60mmol) was dissolved in dry dichloromethane (5mL), and compound II-2 (155mg, 0.30mmol) in dry dichloromethane was slowly added dropwise with stirring (5mL) The solution and triethylamine (84 ⁇ L, 0.60mmol) were reacted at room temperature for 5 hours.
  • the compound III-5 was prepared according to the method of Example 7.
  • the compound III-5 (100mg, 0.145mmol) was dissolved in N,N-dimethylformamide (8mL), and triethylamine (80 ⁇ L, 0.580mmol) was added.
  • 2-(7-oxybenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate 72mg, 0.188mmol
  • compound I after reacting for 1 hour -4 (60mg, 0.174mmol) (prepared in Example 1), the reaction solution was heated to 50°C.
  • the compound I-1 was replaced with I-1' to prepare the compound I-4'.
  • the compound III-2 was prepared according to the method of Example 7.
  • the compound III-2 (150 mg, 0.274 mmol) was dissolved in anhydrous dichloromethane (6 mL), and N,N'-dicyclohexylcarbimide was added in sequence (170 mg, 0.823 mmol), 4-dimethylaminopyridine (34 mg, 0.274 mmol) and monomethyl phthalate (54 mg, 0.302 mmol), the reaction was stirred at room temperature.
  • the compound III-5 was prepared according to the method of Example 7.
  • the compound III-5 120mg, 0.174mmol
  • Morpholine hydrobromide 57 mg, 0.208 mmol
  • potassium carbonate 70 mg, 0.522 mmol
  • the reaction solution was treated with ethyl acetate (20 mL) and (10 mL), the aqueous layer was extracted with ethyl acetate (10 mL), the organic phases were combined, washed with water (10 mL ⁇ 2), and dried with anhydrous sodium sulfate.
  • ESI-MS m/z 527.5 [M+H] + .
  • N,N-dimethylethylenediamine was replaced with D-proline benzyl ester hydrochloride to obtain compound VI-1.
  • the compound VI-1 (120 mg, 0.186 mmol) was dissolved in a mixed solution of tetrahydrofuran (1 mL) and methanol (1 mL), 2N sodium hydroxide solution (1 mL) was added, and the reaction was carried out at 80° C. for 2 hours.
  • N,N-dimethylethylenediamine was replaced with (S)-3-N-benzyloxycarbonylaminopyrrolidine to obtain compound VII-1.
  • compound III-5 was prepared. Dissolve compound III-5 (345mg, 0.500mmol) in anhydrous N,N-dimethylformamide (8mL), add 1,2-dibromoethane (435 ⁇ L, 5.00mmol) and potassium carbonate (70mg , 0.500mmol), the reaction solution was heated to 50°C, and the reaction was stirred for 3 hours.
  • N,N-Dimethylethylenediamine (66 ⁇ L, 0.602mmol) was dissolved in dry dichloromethane (5mL), and compound XI-2 (155mg, 0.301mmol) in dry dichloromethane was slowly added dropwise with stirring (5mL) The solution and triethylamine (84 ⁇ L, 0.602mmol) were reacted at room temperature for 5 hours. After the completion of the reaction detected by TLC, dichloromethane (10mL) was added to dilute the reaction solution, and the reaction solution was washed with saturated sodium bicarbonate solution (10mL ⁇ 3), water (10mL ⁇ 3) and saturated brine (10mL ⁇ 3) successively.
  • N,N-dimethylethylenediamine was replaced with L-proline benzyl ester hydrochloride to prepare compound XII-1.
  • Example 85 4-(2-aminoethyl)thiomorpholine-1,1-dioxide was replaced with N-acetylethylenediamine to prepare compound XVI-1.
  • N-acetylethanolamine was replaced with methanol, and N-(2-aminoethyl)morpholine was replaced with N,N-dimethylethylenediamine to prepare compound B-51:
  • the compound XVII-4 was prepared by referring to the method of Example 106.
  • the compound XVII-4 (300 mg, 0.435 mmol) was dissolved in N,N-dimethylformamide (8 mL), and 1,2-dibromoethane was sequentially added. (375 ⁇ L, 4.348 mmol), potassium carbonate (60 mg, 0.435 mmol), the reaction solution was heated to 50° C., and the reaction was stirred for 3 hours.
  • compound II-1 was prepared. Dissolve compound II-1 (150mg, 0.300mmol) in tetrahydrofuran (8mL), add N,N-diisopropylethylamine (50 ⁇ L, 0.300mmol) and O-benzotriazole-N,N, N',N'-tetramethylurea tetrafluoroboric acid (116mg, 0.361mmol), the reaction was stirred at room temperature. After the completion of the reaction was detected by TLC, the solvent was evaporated under reduced pressure.
  • compound XI-1 was prepared. Dissolve compound XI-1 (150mg, 0.300mmol) in tetrahydrofuran (8mL), add N,N-diisopropylethylamine (50 ⁇ L, 0.300mmol) and O-benzotriazole-N,N, N',N'-tetramethylurea tetrafluoroboric acid (116mg, 0.361mmol), the reaction was stirred at room temperature. After the completion of the reaction was detected by TLC, the solvent was evaporated under reduced pressure.
  • compound B-20 was replaced with compound B-9 to prepare compound B-130: 1 H NMR (300MHz, DMSO-d 6 ) ⁇ 8.53–8.37(m,3H), 5.15– 5.06(m,1H), 4.60-4.50(m,1H), 4.02-3.82(m,5H), 3.18-2.87(m,5H), 1.11(s,3H), 1.01(s,3H), 0.99( s, 3H), 0.90 (s, 3H), 0.87 (s, 6H), 0.66 (s, 3H).
  • ESI-MS m/z 671.5 [M+H] + .
  • the Western Blot method was used to detect the agonistic activity of the compound on Huh-7 cells AMPK.
  • Huh-7 cells human liver cancer cells, purchased from the Stem Cell Bank of the Chinese Academy of Sciences), using DMEM complete medium (containing 10% fetal bovine serum and 1% streptomycin/penicillin), containing 5% CO at 37°C 2. Cultivate in the cell incubator.
  • Antibody anti-AMPK (Cell Signaling Technology 2532S); anti-pAMPK (Thr172, Cell Signaling Technology 2535S); GAPDH (arigobio), HRP-labeled goat anti-rabbit IgG secondary antibody, HRP-labeled goat anti-mouse IgG secondary antibody Anti (Biyuntian).
  • Huh-7 cells are plated at 200,000 per well. Incubate for 24 hours in an incubator containing 5% CO 2 at 37°C, and adhere to the wall.
  • the test compound was administered under the condition of complete medium, the final concentration of the compound was set to 10 ⁇ M, the administration time was 12 hours, and AICAR (100 ⁇ M) was used as the positive control compound.
  • the protein was then extracted for Western Blot detection. Specific steps are as follows:
  • Protein sample preparation discard the original culture medium, wash 3 times with 1 ⁇ PBS, discard PBS, add 100 ⁇ l RIPA buffer (1 ⁇ PBS, 1% NP40, 0.5% sodium deoxycholate, 0.1% SDS, PMSF, etc.) to each well ), incubate on ice for 15 minutes, scrape the cells with a cell scraper, suck into a new 1.5ml EP tube, 4°C, 12000g, centrifuge for 15min, transfer the supernatant to a new 1.5ml EP tube, and place on ice , Add 1/5 volume of 6 ⁇ loading buffer, heat in a metal bath at 95°C for 10min, centrifuge for 1min, freeze and store at -20°C for later use.
  • RIPA buffer 1 ⁇ PBS, 1% NP40, 0.5% sodium deoxycholate, 0.1% SDS, PMSF, etc.
  • Electrophoresis Pouring SDS-PAGE gel, separation gel concentration is 10%, concentrated gel concentration is 4%.
  • the protein samples were heated in a metal bath at 95°C for 4 minutes and then centrifuged for 1 minute. Each sample was loaded with 30 ⁇ g total protein, and the samples were added one by one with a micro-injector. Connect the power supply (note that the positive and negative poles are connected), start electrophoresis at a constant voltage of 60V, when the protein sample enters the separation gel, adjust the voltage to 100V to continue the constant voltage electrophoresis. When bromophenol blue reaches the bottom of the separation gel, the electrophoresis is terminated according to the separation of the protein marker.
  • Transfer membrane Gently take out the gel after electrophoresis, cut off the unnecessary part, and immerse the required gel into the Transfer buffer. Prepare a PVDF membrane with the same size as the glue, soak the PVDF membrane with methanol for 1 min before use, transfer it to the Transfer buffer, and put the filter paper into the Transfer buffer to soak. Spread sponge, filter paper, glue, PVDF membrane, filter paper, and sponge on the electrode plate of the cathode of the transfer apparatus. Avoid air bubbles between each layer. Turn on the power supply and transfer in a 200mA constant current ice bath for 2.5 hours.
  • Antibody hybridization After the transfer, take out the PVDF membrane and wash it with 1 ⁇ TBST once, put it in the pre-prepared blocking solution (1 ⁇ TBS with 0.1% Tween20 in 5% BSA solution), and block at room temperature for 1 hour. Incubate the primary antibody at 4°C overnight. On the second day (after 12h), wash 3 times with 1 ⁇ TBST, each time for 10 minutes. Secondary antibody incubation: Incubate with 5% BSA diluted secondary antibody (1:10000) for 1 hour at room temperature, wash 3 times with 1 ⁇ TBST, 10 minutes each time.
  • the novel derivatives of oleanolic acid and delta-oleanolic acid provided by the present invention have significant AMPK agonistic activity.
  • Both are potent AMPK agonists, and their activity is significantly better than 100 ⁇ M AICAR, oleanolic acid and delta-oleanolic acid.
  • the above experimental results show that the compound of the present invention has significant agonistic activity on AMPK, and therefore can be used to prepare AMPK agonists with the activity of enhancing the phosphorylation level of AMPK, and further can be used to prepare drugs for preventing or treating AMPK-mediated diseases.
  • the synthesis methods of other compounds not listed in the examples of the present invention can be referred to the foregoing examples, and they also have significant agonistic activity on AMPK.
  • Huh-7 cells DMEM complete medium (containing 10% fetal bovine serum and 1% streptomycin/penicillin) cultured in a 5% CO 2 incubator at 37°C.
  • Antibodies anti-ACC (CST, 3676S); anti-pACC (CST, 11818S); anti-mTOR (CST, 2983S); anti-pmTOR (CST, Ser2448).
  • FIG. 1 The experimental results ( Figure 1) show that the compounds of the present invention can effectively cause changes in AMPK downstream proteins, and the change trend is related to AMPK activation.
  • compounds B-125, A-112, B-122, A-109, B-121, A-108, B-123, A-110, B-20 and A-81 can all increase the phosphorylation of AMPK Level, reduce the phosphorylation level of mTOR and increase the phosphorylation level of ACC at the same time.
  • other compounds of the present invention can increase the phosphorylation level of AMPK, reduce the phosphorylation level of mTOR, and increase the phosphorylation level of ACC at the same time.
  • Example 41 The compound A-81 prepared in Example 41 or the compound of other examples (50g), hydroxypropylmethyl cellulose E (150g), starch (200g), a proper amount of povidone K30 and magnesium stearate (1g) ) Mixing, granulating, and tableting.
  • the compounds prepared in Examples 1 to 183 can be given different pharmaceutical excipients into capsules, powders, granules, pills, injections, syrups, oral liquids, inhalants, ointments Agent, suppository or patch, etc.

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Abstract

Disclosed are derivatives of novel pentacyclic triterpenoid AMPK agonists, oleanolic acid and δ-oleanolic acid, and medicinal use thereof, specifically, a compound represented by formula I or formula II, and a pharmaceutically acceptable salt or ester or solvate, which can be used for preparing AMPK agonists having the activity of enhancing an AMPK phosphorylation level and used for preparing a drug for preventing or treating AMPK-mediated diseases. The novel pentacyclic triterpenoid compounds of the present invention have significant AMPK agonist activity, and the activity thereof is significantly better than that of recognized AMPK agonist AICAR, and the compounds also have better pharmacokinetic properties such as oral bioavailability and very good safety.

Description

齐墩果酸和δ-齐墩果酸的衍生物及其医药用途Derivatives of oleanolic acid and δ-oleanolic acid and their medical uses 技术领域Technical field
本发明涉及生物医药领域，涉及具有AMPK激动活性的新型五环三萜类化合物，具体涉及齐墩果酸和δ-齐墩果酸的衍生物及其医药用途，本发明还涉及该类化合物在制备预防或治疗AMPK介导的疾病的药物中的用途及其药物组合物。The present invention relates to the field of biomedicine, relates to novel pentacyclic triterpenoids with AMPK agonistic activity, and specifically relates to derivatives of oleanolic acid and delta-oleanolic acid and their medical uses. The present invention also relates to such compounds in Use in preparing medicines for preventing or treating AMPK-mediated diseases and their pharmaceutical compositions.
背景技术Background technique
AMPK(腺苷酸活化蛋白激酶)是调控机体能量代谢及炎症反应的关键激酶，其磷酸化激活可克服胰岛素抵抗、降血糖、降血脂(通过抑制脂肪酸及胆固醇的合成)、抗炎、抗凋亡、抗纤维化、促进线粒体合成、增强线粒体的氧化代谢、抗衰老和抗肿瘤等(Physiol.Rev.2009,89,1025)。近年来，AMPK的抗炎及抗纤维化作用已越发受到关注(Nature 2013,493,346)，其可能的机制是AMPK通过增强雌激素相关受体α(ERRα)的转录功能而发挥抗炎和抗纤维化作用(Immunity 2015,43,80)。AMPK (Adenylate Activated Protein Kinase) is a key kinase that regulates the body's energy metabolism and inflammation. Its phosphorylation activation can overcome insulin resistance, lower blood sugar, lower blood lipids (by inhibiting the synthesis of fatty acids and cholesterol), anti-inflammatory, and anti-inflammatory Death, anti-fibrosis, promotion of mitochondrial synthesis, enhancement of mitochondrial oxidation metabolism, anti-aging and anti-tumor, etc. (Physiol. Rev. 2009, 89, 1025). In recent years, the anti-inflammatory and anti-fibrotic effects of AMPK have attracted more and more attention (Nature 2013,493,346). The possible mechanism is that AMPK exerts anti-inflammatory and anti-fibrotic functions by enhancing the transcription function of estrogen-related receptor α (ERRα) Chemical effect (Immunity 2015, 43, 80).
越来越多的证据表明，AMPK功能异常与多种疾病的发生发展密切相关。AMPK介导的疾病包括代谢性疾病和心脑血管疾病，如胰岛素抵抗、代谢综合征、1型或2型糖尿病、高脂血症、肥胖症、动脉粥样硬化、心肌缺血、心肌梗死、心律失常、冠心病、高血压、心衰、心肌肥大、心肌炎、糖尿病并发症(包括糖尿病心肌病、糖尿病肾病、视网膜病变、神经病变和糖尿病溃疡等)、非酒精性脂肪肝、非酒精性脂肪性肝炎、酒精性脂肪肝、肝硬化、痛风、中风或脑梗死等；AMPK介导的疾病还包括炎症疾病、自身免疫性疾病、器官纤维化疾病、神经损伤性疾病或病原体感染所致的继发性疾病，如肺炎、哮喘、慢性阻塞性肺病、慢性支气管炎、肺气肿、闭塞性细支气管炎、特发性肺纤维化、囊性纤维化肺病、过敏性鼻炎、炎性肠病(如克罗恩病和溃疡性结肠炎)、多囊肾病、多囊卵巢综合征(PCOS)、白塞氏病、系统性红斑狼疮、类风湿关节炎、脊椎关节炎、骨关节炎、滑膜炎、肌腱炎、血栓闭塞性脉管炎、静脉炎、间歇性跛行、瘢痕瘤、银屑病、鱼鳞癣、大疱性类天疱疮、皮炎、接触性皮炎、胰腺炎、慢性肾炎、膀胱炎、脑膜炎、胃炎、败血症、坏疽性脓皮症、葡萄膜炎、帕金森病、阿尔茨海默病、α-共核蛋白病、抑郁症、多发性硬化症、肌萎缩侧索硬化病、纤维肌痛综合症、神经痛、唐氏综合征、哈勒沃登-施帕病、亨廷顿舞蹈病或威尔逊病等。More and more evidences show that AMPK dysfunction is closely related to the occurrence and development of many diseases. AMPK-mediated diseases include metabolic diseases and cardiovascular and cerebrovascular diseases, such as insulin resistance, metabolic syndrome, type 1 or type 2 diabetes, hyperlipidemia, obesity, atherosclerosis, myocardial ischemia, myocardial infarction, Arrhythmia, coronary heart disease, hypertension, heart failure, myocardial hypertrophy, myocarditis, diabetic complications (including diabetic cardiomyopathy, diabetic nephropathy, retinopathy, neuropathy and diabetic ulcers, etc.), non-alcoholic fatty liver, non-alcoholic fat Hepatitis, alcoholic fatty liver, cirrhosis, gout, stroke or cerebral infarction, etc.; AMPK-mediated diseases also include inflammatory diseases, autoimmune diseases, organ fibrosis diseases, neurological damage diseases, or subsequent infections caused by pathogens Primary diseases, such as pneumonia, asthma, chronic obstructive pulmonary disease, chronic bronchitis, emphysema, bronchiolitis obliterans, idiopathic pulmonary fibrosis, cystic fibrotic lung disease, allergic rhinitis, inflammatory bowel disease ( Such as Crohn's disease and ulcerative colitis), polycystic kidney disease, polycystic ovary syndrome (PCOS), Behcet's disease, systemic lupus erythematosus, rheumatoid arthritis, spondyloarthritis, osteoarthritis, synovium Inflammation, tendinitis, thromboangiitis obliterans, phlebitis, intermittent claudication, keloids, psoriasis, ichthyosis, bullous pemphigoid, dermatitis, contact dermatitis, pancreatitis, chronic nephritis, bladder Inflammation, meningitis, gastritis, sepsis, pyoderma gangrenosum, uveitis, Parkinson's disease, Alzheimer's disease, alpha-synucleinopathy, depression, multiple sclerosis, amyotrophic lateral sclerosis , Fibromyalgia Syndrome, Neuralgia, Down Syndrome, Hallerwarden-Spar disease, Huntington’s disease or Wilson’s disease.
文献报道AMPK激动剂可预防和治疗多种AMPK介导的疾病(J.Med.Chem.,2015,58 2；Nature 2013,493,346；Experimental Neurology 2017,298,31； Biochemical Pharmacology 2010,80,1708；Current Drug Targets,2016,17,908；Nat Rev Drug Discov,2019,DOI:10.1038/s41573-019-0019-2)。例如，临床上广泛使用的降糖药二甲双胍认为主要是通过激活AMPK而发挥多种临床功效(J.Clin.Invest.2001,108,1167)。尽管AMPK激动剂具有广泛的临床应用前景，然而，迄今为止，研制新型安全、有效的AMPK激动剂尚未取得实质性进展。由于安全性或有效性的原因，在研的AMPK激动剂进入临床研究阶段的寥寥无几。例如，广谱AMPK-β亚基激动剂MK-8722尽管可以降血糖，但在大鼠和猴子实验中发现动物心脏发生不可逆性心肌肥厚副作用(Science 2017,357,507)。此外，AICAR作为最常用的AMPK激动剂之一(Eur.J.Biochem.1995,229,558)，也因其较大的毒副作用而被终止了临床实验(J Clin Pharmacol 1991,31:342–347)。It is reported in the literature that AMPK agonists can prevent and treat a variety of AMPK-mediated diseases (J.Med.Chem.,2015,58 2; Nature 2013,493,346; Experimental Neurology 2017,298,31; Biochemical Pharmacology 2010,80,1708; Current Drug Targets, 2016, 17, 908; Nat Rev Drug Discov, 2019, DOI: 10.1038/s41573-019-0019-2). For example, metformin, a hypoglycemic drug widely used clinically, is believed to exert a variety of clinical effects by activating AMPK (J. Clin. Invest. 2001, 108, 1167). Although AMPK agonists have broad clinical application prospects, so far, no substantial progress has been made in the development of new safe and effective AMPK agonists. Due to safety or effectiveness reasons, very few AMPK agonists under development have entered the clinical research phase. For example, although the broad-spectrum AMPK-β subunit agonist MK-8722 can lower blood sugar, it has been found in rat and monkey experiments that the animal heart has the side effects of irreversible cardiac hypertrophy (Science 2017, 357, 507). In addition, AICAR, as one of the most commonly used AMPK agonists (Eur. J. Biochem. 1995, 229, 558), has also been terminated in clinical trials due to its relatively large side effects (J Clin Pharmacol 1991, 31: 342-347) .
总之，临床上亟需开发活性高、毒副作用小的新型AMPK激动剂。另一方面，齐墩果酸是药用植物中常见的一个五环三萜，其具有广泛的生物活性(Nat Prod Rep 2011,28,543)。而3β-羟基-齐墩果烷-13(18)-烯-28-酸(δ-齐墩果酸)是一个在自然界中非常罕见的五环三萜酸(Phytochemistry 1999,51,83)。迄今，极少有文献报道δ-齐墩果酸的生物活性。In short, there is an urgent need to develop new AMPK agonists with high activity and low side effects. On the other hand, oleanolic acid is a common pentacyclic triterpene in medicinal plants, which has a wide range of biological activities (Nat Prod Rep 2011, 28, 543). And 3β-hydroxy-oleanan-13(18)-en-28-acid (δ-oleanolic acid) is a pentacyclic triterpene acid that is very rare in nature (Phytochemistry 1999, 51, 83). So far, there are very few reports on the biological activity of δ-oleanolic acid.
发明内容Summary of the invention
发明目的：针对AMPK激动剂研发领域现有技术存在的问题，本发明提供一种新型齐墩果酸和δ-齐墩果酸的衍生物；本发明提供的新型齐墩果酸衍生物或者δ-齐墩果酸衍生物是一种新型的AMPK激动剂，因而可用于制备预防或治疗AMPK介导的疾病的药物。Objective of the invention: In view of the problems existing in the prior art in the field of AMPK agonist research and development, the present invention provides a novel oleanolic acid and δ-oleanolic acid derivatives; the new oleanolic acid derivative or δ -Oleanolic acid derivatives are a new type of AMPK agonist, so they can be used to prepare drugs for preventing or treating AMPK-mediated diseases.
本发明人在对δ-齐墩果酸和齐墩果酸进行结构改造时，意外地发现了一系列强效的新型AMPK激动剂，且其AMPK激动活性显著地优于公认的AMPK激动剂AICAR。The inventors unexpectedly discovered a series of potent new AMPK agonists when they modified the structure of δ-oleanolic acid and oleanolic acid, and their AMPK agonist activity was significantly better than the recognized AMPK agonist AICAR .
技术方案：为了实现上述目的，如本发明所述如下式I或式II所示的δ-齐墩果酸衍生物或齐墩果酸衍生物、其药学上可接受的盐或酯或溶剂化物：Technical solution: In order to achieve the above object, as described in the present invention, the δ-oleanolic acid derivative or the oleanolic acid derivative represented by the following formula I or formula II, and a pharmaceutically acceptable salt or ester or solvate thereof :
Figure PCTCN2020093248-appb-000001
Figure PCTCN2020093248-appb-000001
其中，among them,
R是H、
Figure PCTCN2020093248-appb-000002
或R aCO-； R is H,
Figure PCTCN2020093248-appb-000002
Or R a CO-;
R 1是H、C 1-C 5烷基或取代基Y取代的C 1-C 5烷基，所述取代基Y是OH、C(O)OH、C(O)NH 2、NH 2、NHC(O)CH 3、吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基或二乙醇胺基； R 1 is H, C 1 -C 5 alkyl substituted by substituent Y or a C 1 -C 5 alkyl, the substituent Y is OH, C (O) OH, C (O) NH 2, NH 2, NHC(O)CH 3 , pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorph Lin-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethylammonium or diethanolamino;
R a是非取代的或取代基L取代的C 1-C 5烷基，所述取代基L是一个或两个独立地选自下列的取代基：OH、C(O)OH、NH 2、吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基或NHC(O)CH(CH 3)NHC(O)CH(CH 3)NH； R a is an unsubstituted or substituted group L is substituted C 1 -C 5 alkyl, the substituent group L is one or two substituents independently selected from the group: OH, C (O) OH , NH 2, pyrrolidin Alk-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorpholine-1,1-diox Generation-4-yl or NHC(O)CH(CH 3 )NHC(O)CH(CH 3 )NH;
R’是OR 2、NR 3R 4或
Figure PCTCN2020093248-appb-000003
R'is OR 2 , NR 3 R 4 or
Figure PCTCN2020093248-appb-000003
R 2是取代基Z取代的C 1-C 5烷基，所述取代基Z是吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基或二乙醇胺基； R 2 is a C 1 -C 5 alkyl group substituted by a substituent Z, and the substituent Z is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium or diethanolamine group;
R 3是H或C 1-C 3烷基； R 3 is H or C 1 -C 3 alkyl;
R 4是取代基W取代的C 1-C 5烷基，所述取代基W是吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基、二乙醇胺基或乙酰基氨基； R 4 is a C 1 -C 5 alkyl group substituted by a substituent W, the substituent W is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium, diethanolamine or acetylamino;
R 5是连接在环上任一碳上的H、OH、F、NH 2、C 1-C 3烷基氨基、COOH、C 1-C 3烷基、C 1-C 3烷氧基、环烷基氧基或杂环烷基氧基； R 5 is H, OH, F, NH 2 , C 1 -C 3 alkylamino, COOH, C 1 -C 3 alkyl, C 1 -C 3 alkoxy, cycloalkane attached to any carbon on the ring Oxy or heterocycloalkyloxy;
Q是CH 2、O、NR 6、S、SO 2、CHR 7或化学键； Q is CH 2 , O, NR 6 , S, SO 2 , CHR 7 or chemical bond;
R 6是H、C 1-C 3烷基、C 1-C 3烷基酰基、C 1-C 3烷基磺酰基或取代基P取代的C 1-C 3烷基，所述取代基P是OH、COOH、NH 2或C 1-C 3烷基氨基； R 6 is H, C 1 -C 3 alkyl, C 1 -C 3 alkyl acyl, C 1 -C 3 alkylsulfonyl, or C 1 -C 3 alkyl substituted by the substituent P, the substituent P Is OH, COOH, NH 2 or C 1 -C 3 alkylamino;
R 7是OH、NH 2、COOH、C 1-C 3烷基氨基、C 1-C 3烷氧基、环烷基氧基或杂环烷基氧基； R 7 is OH, NH 2 , COOH, C 1 -C 3 alkylamino, C 1 -C 3 alkoxy, cycloalkyloxy or heterocycloalkyloxy;
n是0、1或2；n is 0, 1 or 2;
m是0、1或2。m is 0, 1, or 2.
在某些优选的实施方案中，所述的式I或式II化合物、其药学上可接受的盐或酯或溶剂化物，In certain preferred embodiments, the compound of Formula I or Formula II, or a pharmaceutically acceptable salt or ester or solvate thereof,
R是H、
Figure PCTCN2020093248-appb-000004
或R aCO-； R is H,
Figure PCTCN2020093248-appb-000004
Or R a CO-;
R 1是H、C 1-C 5烷基或取代基Y取代的C 1-C 5烷基，所述取代基Y是OH、C(O)OH、C(O)NH 2、NH 2、NHC(O)CH 3、吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基或二乙醇胺基； R 1 is H, C 1 -C 5 alkyl substituted by substituent Y or a C 1 -C 5 alkyl, the substituent Y is OH, C (O) OH, C (O) NH 2, NH 2, NHC(O)CH 3 , pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorph Lin-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethylammonium or diethanolamino;
R a是非取代的或取代基L取代的C 1-C 5烷基，所述取代基L是一个或两个独立地选自下列的取代基：OH、C(O)OH、NH 2、吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基或NHC(O)CH(CH 3)NHC(O)CH(CH 3)NH； R a is an unsubstituted or substituted group L is substituted C 1 -C 5 alkyl, the substituent group L is one or two substituents independently selected from the group: OH, C (O) OH , NH 2, pyrrolidin Alk-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorpholine-1,1-diox Generation-4-yl or NHC(O)CH(CH 3 )NHC(O)CH(CH 3 )NH;
R’是OR 2、NR 3R 4或
Figure PCTCN2020093248-appb-000005
R'is OR 2 , NR 3 R 4 or
Figure PCTCN2020093248-appb-000005
R 2是取代基Z取代的C 1-C 5烷基，所述取代基Z是吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基或二乙醇胺基；R 3是H； R 2 is a C 1 -C 5 alkyl group substituted by a substituent Z, and the substituent Z is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium or diethanolamine; R 3 is H;
R 4是取代基W取代的C 1-C 5烷基，所述取代基W是吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基、二乙醇胺基或乙酰基氨基； R 4 is a C 1 -C 5 alkyl group substituted by a substituent W, the substituent W is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium, diethanolamine or acetylamino;
R 5是连接在环上任一碳上的H、OH、F、NH 2、C 1-C 3烷基氨基或COOH； R 5 is H, OH, F, NH 2 , C 1 -C 3 alkylamino or COOH attached to any carbon on the ring;
Q是CH 2、O、NR 6、S、SO 2或CHR 7； Q is CH 2 , O, NR 6 , S, SO 2 or CHR 7 ;
R 6是H、C 1-C 3烷基、C 1-C 3烷基酰基、C 1-C 3烷基磺酰基或取代基P取代的C 1-C 3烷基，所述取代基P是OH、COOH、NH 2或C 1-C 3烷基氨基； R 6 is H, C 1 -C 3 alkyl, C 1 -C 3 alkyl acyl, C 1 -C 3 alkylsulfonyl or C 1 -C 3 alkyl substituted with substituent P, said substituent P Is OH, COOH, NH 2 or C 1 -C 3 alkylamino;
R 7是OH、NH 2、COOH或C 1-C 3烷基氨基； R 7 is OH, NH 2 , COOH or C 1 -C 3 alkylamino;
n是0或1；n is 0 or 1;
m是0或1。m is 0 or 1.
更进一步地，在所述式II化合物中，当取代基Z是N,N-二甲基氨基或三甲基铵基时，R不是H；当取代基W是乙酰基氨基时，R不是H。Furthermore, in the compound of formula II, when the substituent Z is N,N-dimethylamino or trimethylammonium, R is not H; when the substituent W is acetylamino, R is not H .
在某些更优选的实施方案中，所述化合物或其药学上可接受的盐或酯或溶剂化物选自如下化合物：In certain more preferred embodiments, the compound or a pharmaceutically acceptable salt or ester or solvate thereof is selected from the following compounds:
Figure PCTCN2020093248-appb-000006
Figure PCTCN2020093248-appb-000006
Figure PCTCN2020093248-appb-000007
Figure PCTCN2020093248-appb-000007
Figure PCTCN2020093248-appb-000008
Figure PCTCN2020093248-appb-000008
Figure PCTCN2020093248-appb-000009
Figure PCTCN2020093248-appb-000009
Figure PCTCN2020093248-appb-000010
Figure PCTCN2020093248-appb-000010
Figure PCTCN2020093248-appb-000011
Figure PCTCN2020093248-appb-000011
Figure PCTCN2020093248-appb-000012
Figure PCTCN2020093248-appb-000012
Figure PCTCN2020093248-appb-000013
Figure PCTCN2020093248-appb-000013
Figure PCTCN2020093248-appb-000014
Figure PCTCN2020093248-appb-000014
Figure PCTCN2020093248-appb-000015
Figure PCTCN2020093248-appb-000015
Figure PCTCN2020093248-appb-000016
Figure PCTCN2020093248-appb-000016
Figure PCTCN2020093248-appb-000017
Figure PCTCN2020093248-appb-000017
Figure PCTCN2020093248-appb-000018
Figure PCTCN2020093248-appb-000018
Figure PCTCN2020093248-appb-000019
Figure PCTCN2020093248-appb-000019
Figure PCTCN2020093248-appb-000020
Figure PCTCN2020093248-appb-000020
Figure PCTCN2020093248-appb-000021
Figure PCTCN2020093248-appb-000021
Figure PCTCN2020093248-appb-000022
Figure PCTCN2020093248-appb-000022
Figure PCTCN2020093248-appb-000023
Figure PCTCN2020093248-appb-000023
Figure PCTCN2020093248-appb-000024
Figure PCTCN2020093248-appb-000024
Figure PCTCN2020093248-appb-000025
Figure PCTCN2020093248-appb-000025
Figure PCTCN2020093248-appb-000026
Figure PCTCN2020093248-appb-000026
Figure PCTCN2020093248-appb-000027
Figure PCTCN2020093248-appb-000027
Figure PCTCN2020093248-appb-000028
Figure PCTCN2020093248-appb-000028
Figure PCTCN2020093248-appb-000029
Figure PCTCN2020093248-appb-000029
Figure PCTCN2020093248-appb-000030
Figure PCTCN2020093248-appb-000030
Figure PCTCN2020093248-appb-000031
Figure PCTCN2020093248-appb-000031
Figure PCTCN2020093248-appb-000032
Figure PCTCN2020093248-appb-000032
Figure PCTCN2020093248-appb-000033
Figure PCTCN2020093248-appb-000033
Figure PCTCN2020093248-appb-000034
Figure PCTCN2020093248-appb-000034
Figure PCTCN2020093248-appb-000035
Figure PCTCN2020093248-appb-000035
本发明的化合物也可作为药用盐使用。该盐可为下列酸中的至少一种的酸盐：半乳糖二酸、D-葡糖醛酸、甘油磷酸、马尿酸、羟乙磺酸、乳糖酸、马来酸、1,5-萘二磺酸、萘-2-磺酸、新戊酸、对苯二甲酸、硫氰酸、胆酸、正十二烷基硫酸、苯磺酸、柠檬酸、D-葡萄糖，乙醇酸、乳酸、苹果酸、丙二酸、扁桃酸、磷酸、丙酸、盐酸、硫酸、酒石酸、琥珀酸、甲酸、氢碘酸、氢溴酸、甲烷磺酸、 烟酸、硝酸、乳清酸、草酸、苦味酸、L-焦谷氨酸、糖精酸、水杨酸、龙胆酸、对甲苯磺酸、戊酸、棕榈酸、葵二酸、硬脂酸、月桂酸、乙酸、己二酸、碳酸、苯磺酸、乙烷二磺酸、乙基琥珀酸、富马酸、3-羟基萘-2-甲酸、1-羟基萘-2-甲酸、油酸、十一碳烯酸、抗坏血酸、樟脑酸、樟脑磺酸、二氯乙酸、乙烷磺酸。另一方面，该盐也可以是本发明的化合物与金属(包括钠、钾、钙等)离子或药学上可接受的胺(包括乙二胺、氨丁三醇等)、铵离子或胆碱形成的盐。本发明的化合物也可以按酯、前药形式、N-氧化物或其溶剂化物组成药物组合物。The compounds of the present invention can also be used as pharmaceutical salts. The salt may be the acid salt of at least one of the following acids: galactonic acid, D-glucuronic acid, glycerophosphoric acid, hippuric acid, isethionic acid, lactobionic acid, maleic acid, 1,5-naphthalene Disulfonic acid, naphthalene-2-sulfonic acid, pivalic acid, terephthalic acid, thiocyanic acid, cholic acid, n-dodecyl sulfuric acid, benzenesulfonic acid, citric acid, D-glucose, glycolic acid, lactic acid, Malic acid, malonic acid, mandelic acid, phosphoric acid, propionic acid, hydrochloric acid, sulfuric acid, tartaric acid, succinic acid, formic acid, hydroiodic acid, hydrobromic acid, methanesulfonic acid, niacin, nitric acid, orotic acid, oxalic acid, bitter Acid, L-pyroglutamic acid, saccharic acid, salicylic acid, gentisic acid, p-toluenesulfonic acid, valeric acid, palmitic acid, sebacic acid, stearic acid, lauric acid, acetic acid, adipic acid, carbonic acid, Benzenesulfonic acid, ethanedisulfonic acid, ethylsuccinic acid, fumaric acid, 3-hydroxynaphthalene-2-carboxylic acid, 1-hydroxynaphthalene-2-carboxylic acid, oleic acid, undecylenic acid, ascorbic acid, camphor acid , Camphor sulfonic acid, dichloroacetic acid, ethane sulfonic acid. On the other hand, the salt can also be the compound of the present invention and metal (including sodium, potassium, calcium, etc.) ions or pharmaceutically acceptable amines (including ethylenediamine, tromethamine, etc.), ammonium ions or choline The salt formed. The compounds of the present invention can also be formed into pharmaceutical compositions in the form of esters, prodrugs, N-oxides or their solvates.
齐墩果酸可商业化购买。δ-齐墩果酸的合成可参照文献方法(Org.Biomol.Chem.,2016,14,11154)进行，即首先参照文献方法制得δ-齐墩果酸的苄酯，再经过常规的催化氢化脱苄反应即可制得δ-齐墩果酸。δ-齐墩果酸和齐墩果酸衍生物的合成可参照实施例的方法或改进的方法进行。Oleanolic acid can be purchased commercially. The synthesis of δ-oleanolic acid can be carried out with reference to the literature method (Org.Biomol.Chem., 2016, 14, 11154), that is, the benzyl ester of δ-oleanolic acid is prepared by referring to the literature method, and then subjected to conventional catalysis Hydrogenation debenzylation can produce δ-oleanolic acid. The synthesis of δ-oleanolic acid and oleanolic acid derivatives can be carried out by referring to the method in the examples or the improved method.
本发明提供了所述的式I或式II化合物、其药学上可接受的盐或酯或溶剂化物在制备具有增强AMPK磷酸化水平活性的AMPK激动剂中的应用。本发明的化合物对AMPK具有显著的激动活性，因而可用于制备具有增强AMPK磷酸化水平活性的AMPK激动剂。The present invention provides the use of the compound of formula I or formula II, its pharmaceutically acceptable salt or ester or solvate in the preparation of AMPK agonists with the activity of enhancing AMPK phosphorylation level. The compound of the present invention has significant agonistic activity on AMPK, and therefore can be used to prepare AMPK agonists with activity to enhance AMPK phosphorylation level.
本发明还提供了所述的式I或式II化合物、其药学上可接受的盐或酯或溶剂化物在制备预防或治疗AMPK介导的疾病的药物中的用途。The present invention also provides the use of the compound of formula I or formula II, its pharmaceutically acceptable salt or ester or solvate in the preparation of a medicine for preventing or treating AMPK-mediated diseases.
其中，所述AMPK介导的疾病包括代谢性疾病、心脑血管疾病、炎症疾病、自身免疫性疾病、器官纤维化疾病、神经退行性疾病、病原体感染所致的继发性疾病、线粒体功能障碍或紊乱疾病或肿瘤。Wherein, the AMPK-mediated diseases include metabolic diseases, cardiovascular and cerebrovascular diseases, inflammatory diseases, autoimmune diseases, organ fibrotic diseases, neurodegenerative diseases, secondary diseases caused by pathogen infection, and mitochondrial dysfunction Or disorder disease or tumor.
所述AMPK介导的疾病，如代谢性疾病和心脑血管疾病，包括：如胰岛素抵抗、代谢综合征、1型或2型糖尿病、高脂血症、肥胖症、动脉粥样硬化、心肌缺血、心肌梗死、心律失常、冠心病、高血压、心衰、心肌肥大、心肌炎、糖尿病并发症(包括糖尿病心肌病、糖尿病肾病、视网膜病变、神经病变和糖尿病溃疡等)、非酒精性脂肪肝、非酒精性脂肪性肝炎、酒精性脂肪肝、肝硬化、痛风、中风或脑梗死等。The AMPK-mediated diseases, such as metabolic diseases and cardiovascular and cerebrovascular diseases, include: insulin resistance, metabolic syndrome, type 1 or type 2 diabetes, hyperlipidemia, obesity, atherosclerosis, myocardial deficiency Blood, myocardial infarction, arrhythmia, coronary heart disease, hypertension, heart failure, myocardial hypertrophy, myocarditis, diabetic complications (including diabetic cardiomyopathy, diabetic nephropathy, retinopathy, neuropathy, diabetic ulcers, etc.), non-alcoholic fatty liver , Non-alcoholic steatohepatitis, alcoholic fatty liver, cirrhosis, gout, stroke or cerebral infarction, etc.
所述AMPK介导的疾病，如炎症疾病、自身免疫性疾病、器官纤维化疾病、神经损伤性疾病或病原体感染所致的继发性疾病，包括：肺炎、哮喘、慢性阻塞性肺病、慢性支气管炎、肺气肿、闭塞性细支气管炎、特发性肺纤维化、囊性纤维化肺病、过敏性鼻炎、炎性肠病(如克罗恩病和溃疡性结肠炎)、多囊肾病、多囊卵巢综合征(PCOS)、白塞氏病、系统性红斑狼疮、类风湿关节炎、脊椎关节炎、骨关节炎、滑膜炎、肌腱炎、血栓闭塞性脉管炎、静脉炎、间歇性跛行、瘢痕瘤、银屑病、鱼鳞癣、大疱性类天疱疮、皮炎、接触性皮炎、胰腺炎、慢性 肾炎、膀胱炎、脑膜炎、胃炎、败血症、坏疽性脓皮症、葡萄膜炎、帕金森病、阿尔茨海默病、α-共核蛋白病、抑郁症、多发性硬化症、肌萎缩侧索硬化病、纤维肌痛综合症、神经痛、唐氏综合征、哈勒沃登-施帕病、亨廷顿舞蹈病或威尔逊病等。The AMPK-mediated diseases, such as inflammatory diseases, autoimmune diseases, organ fibrotic diseases, neurological damage diseases, or secondary diseases caused by pathogen infection, include: pneumonia, asthma, chronic obstructive pulmonary disease, chronic bronchus Inflammation, emphysema, bronchiolitis obliterans, idiopathic pulmonary fibrosis, cystic fibrotic lung disease, allergic rhinitis, inflammatory bowel disease (such as Crohn’s disease and ulcerative colitis), polycystic kidney disease, Polycystic ovary syndrome (PCOS), Behcet's disease, systemic lupus erythematosus, rheumatoid arthritis, spondyloarthritis, osteoarthritis, synovitis, tendinitis, thromboangiitis obliterans, phlebitis, intermittent Claudication, keloid, psoriasis, ichthyosis, bullous pemphigoid, dermatitis, contact dermatitis, pancreatitis, chronic nephritis, cystitis, meningitis, gastritis, sepsis, pyoderma gangrenosum, grape Meningitis, Parkinson's disease, Alzheimer's disease, α-synuclein disease, depression, multiple sclerosis, amyotrophic lateral sclerosis, fibromyalgia syndrome, neuralgia, Down syndrome, Ha Leverden-Spar disease, Huntington's disease or Wilson disease, etc.
所述AMPK介导的疾病，如线粒体功能障碍和紊乱疾病，包括：肌无力、肌阵挛、运动不耐受、卡恩斯-赛尔综合征、慢性疲乏综合征、利氏综合征、线粒体肌病-脑病-高乳酸血症、中风综合征或中风样发作。同样，本发明的化合物也可用于治疗肌肉营养不良状态，例如，杜氏肌营养不良、贝壳肌营养不良或弗立德希氏共济失调。The AMPK-mediated diseases, such as mitochondrial dysfunction and disorder, include: myasthenia, myoclonus, exercise intolerance, Karnes-Saier syndrome, chronic fatigue syndrome, Leigh syndrome, mitochondria Myopathy-encephalopathy-hyperlactic acidemia, stroke syndrome or stroke-like episodes. Similarly, the compounds of the present invention can also be used to treat muscular dystrophy conditions, for example, Duchenne muscular dystrophy, conch muscular dystrophy, or Friedrich's ataxia.
所述AMPK介导的疾病，如肿瘤，包括：骨癌、急性骨髓性白血病、慢性骨髓性白血病、急性淋巴细胞系白血病、慢性淋巴细胞系白血病、骨髓增生性疾病、多发性骨髓瘤、骨髓增生异常综合征、霍奇金氏淋巴瘤、非霍奇金氏淋巴瘤、血管瘤、肉芽瘤、黄瘤、脑膜肉瘤、神经胶质瘤、星形细胞瘤、成神经管细胞瘤、室管膜瘤、生殖细胞瘤(松果体瘤)、多形性成胶质细胞瘤、少突神经胶质细胞瘤、神经鞘瘤、成视网膜细胞瘤、纤维神经瘤、肉瘤、食道癌、胃癌、胰腺癌、大肠癌、结肠癌、直肠癌、肾癌、前列腺癌、淋巴癌、睾丸癌、间质细胞癌、肺癌、肝癌、皮肤癌、恶性黑素瘤或基底细胞癌等。The AMPK-mediated diseases, such as tumors, include: bone cancer, acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, and myeloproliferation Abnormal syndrome, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hemangioma, granuloma, xanthoma, meningiosarcoma, glioma, astrocytoma, medulloblastoma, ependymal Tumor, germ cell tumor (pineal tumor), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, fibroneuronoma, sarcoma, esophageal cancer, gastric cancer, pancreas Cancer, colorectal cancer, colon cancer, rectal cancer, kidney cancer, prostate cancer, lymphoma, testicular cancer, stromal cell carcinoma, lung cancer, liver cancer, skin cancer, malignant melanoma or basal cell carcinoma, etc.
本发明所述一种预防或治疗AMPK介导的疾病的药物组合物，其中含有治疗有效量的式I或式II化合物或其药学上可接受的盐或酯或溶剂化物作为活性成份和药学上可接受的辅料。可任意混合的辅料根据剂型、给药形式等可以改变。辅料的例子包括赋形剂、粘合剂、崩解剂、润滑剂、矫味剂、香味剂、着色剂或甜味剂等。所述药物组合物可以是胶囊剂、散剂、片剂、颗粒剂、丸剂、注射剂、糖浆剂、口服液、吸入剂、软膏剂、栓剂或贴剂等制剂学上常规的制剂形式。The pharmaceutical composition for preventing or treating AMPK-mediated diseases of the present invention contains a therapeutically effective amount of a compound of formula I or formula II or a pharmaceutically acceptable salt or ester or solvate thereof as an active ingredient and a pharmaceutical composition Acceptable excipients. The adjuvants that can be mixed arbitrarily can be changed according to the dosage form and administration form. Examples of excipients include excipients, binders, disintegrants, lubricants, correctives, flavors, coloring agents, or sweeteners. The pharmaceutical composition can be in the form of capsules, powders, tablets, granules, pills, injections, syrups, oral liquids, inhalants, ointments, suppositories or patches, etc.
如果需要，本发明的式I或式II化合物或其药学上可接受的盐或酯或溶剂化物可与一种或多种其他类型的预防或治疗AMPK介导的疾病的药物联合使用，包括但不限于以下几种联合用药的情形。If necessary, the compound of formula I or formula II of the present invention or a pharmaceutically acceptable salt or ester or solvate thereof can be used in combination with one or more other types of drugs for preventing or treating AMPK-mediated diseases, including but It is not limited to the following combinations of drugs.
可选择与本发明的化合物联合使用的药物可以是一种或多种抗糖尿病药物，包括二甲双胍、磺酰脲类降糖药(如格列苯脲和格列美脲等)、葡萄糖苷酶抑制剂(如阿卡波糖和米格列醇等)、PPARγ激动剂(如吡格列酮和罗格列酮)、PPARα/γ双重激动剂、二肽基肽酶IV(DPP-IV)抑制剂(如西格列汀、沙格列汀、阿格列汀和利格列汀等)、格列奈类降糖药(如瑞格列奈和那格列奈等)、SGLT2抑制剂(如坎格列净、达格列净、恩格列净、依格列净、鲁格列净和托格列净等)、葡萄糖激酶激动剂(如HMS5552等)、胰岛素、胰高血糖素样肽-1(GLP-1)类药 物(如埃塞那肽、利拉鲁肽、利司那肽、杜拉鲁肽、贝那鲁肽和阿必鲁肽等)、PTP1B抑制剂、糖原磷酸化酶抑制剂、葡萄糖-6-磷酸酶抑制剂、AMPK激动剂、GPR40激动剂或GPR120激动剂。The drugs that can be used in combination with the compounds of the present invention can be one or more anti-diabetic drugs, including metformin, sulfonylurea hypoglycemic drugs (such as glibenclamide and glimepiride, etc.), glucosidase inhibitors (Such as acarbose and miglitol), PPARγ agonists (such as pioglitazone and rosiglitazone), PPARα/γ dual agonists, dipeptidyl peptidase IV (DPP-IV) inhibitors (such as Sitagliptin, saxagliptin, alogliptin and linagliptin, etc.), Glinide-type hypoglycemic drugs (such as repaglinide and nateglinide, etc.), SGLT2 inhibitors (such as Kangel Liegliflozin, Dapagliflozin, Enpagliflozin, Igligliflozin, Rupagliflozin and Togliflozin, etc.), glucokinase agonists (such as HMS5552, etc.), insulin, glucagon-like peptide-1 (GLP-1) drugs (such as exenatide, liraglutide, risnatide, duraglutide, benaglutide and abiglutide, etc.), PTP1B inhibitors, glycogen phosphorylase Inhibitor, glucose-6-phosphatase inhibitor, AMPK agonist, GPR40 agonist, or GPR120 agonist.
可选择与本发明的化合物联合使用的药物可以是一种或多种减肥药物，包括氯卡色林、奥利司他和胰高血糖素样肽-1(GLP-1)类药物(如埃塞那肽、利拉鲁肽、利司那肽、杜拉鲁肽、贝那鲁肽和阿必鲁肽等)等。The drugs that can be used in combination with the compounds of the present invention can be one or more weight loss drugs, including lorcaserin, orlistat, and glucagon-like peptide-1 (GLP-1) drugs (such as A Sernatide, liraglutide, lisnatide, duraglutide, benaglutide and abiglutide, etc.).
可选择与本发明的化合物联合使用的药物可以是一种或多种抗非酒精性脂肪性肝病药物，包括：AMPK激动剂(如二甲双胍)、法尼酯X受体(FXR)激动剂(如奥贝胆酸、GS-9674、EDP-305和LJN452等)、乙酰辅酶A羧化酶(ACC)抑制剂(如GS-0976等)、凋亡信号调节激酶-1(ASK1)抑制剂(如Selonsertib等)、PPAR激动剂(如Elafibranor、Saroglitazar、IVA337和MSDC-0602K等)、半胱天冬酶(caspase)抑制剂(如Emricasan等)、硬脂酰辅酶A去饱和酶1(SCD1)抑制剂(如Aramchol等)、长效胰高血糖素样肽-1(GLP-1)受体激动剂(如Semaglutide等)、顶端钠依赖性胆盐转运体(ASBT)抑制剂(如Volixibat等)、血管粘附蛋白1(VAP-1)抑制剂(如BI 1467335等)、CCR5R阻断剂(如Cenicriviroc等)和甲状腺激素受体β(THR-β)激动剂(如MGL-3196等)等。The drugs that can be used in combination with the compounds of the present invention can be one or more anti-non-alcoholic fatty liver disease drugs, including: AMPK agonists (such as metformin), farnesoid X receptor (FXR) agonists (such as Obeticholic acid, GS-9674, EDP-305 and LJN452, etc.), acetyl-Coenzyme A carboxylase (ACC) inhibitors (such as GS-0976, etc.), apoptosis signal-regulated kinase-1 (ASK1) inhibitors (such as Selonsertib, etc.), PPAR agonists (such as Elafibranor, Saroglitazar, IVA337 and MSDC-0602K, etc.), caspase inhibitors (such as Emricasan, etc.), stearoyl-CoA desaturase 1 (SCD1) inhibition Agents (such as Aramchol, etc.), long-acting glucagon-like peptide-1 (GLP-1) receptor agonists (such as Semaglutide, etc.), apical sodium-dependent bile salt transporter (ASBT) inhibitors (such as Volixibat, etc.) , Vascular adhesion protein 1 (VAP-1) inhibitors (such as BI 1467335, etc.), CCR5R blockers (such as Cenicriviroc, etc.), and thyroid hormone receptor β (THR-β) agonists (such as MGL-3196, etc.), etc. .
可选择与本发明的化合物联合使用的药物可以是一种或多种降血脂药物，包括烟酸、他汀类药物(如洛伐他丁、辛伐他汀、普伐他汀、美伐他汀、氟伐他汀、阿托伐他汀、西立伐他汀、罗伐他汀和pitavastatin)、胆固醇吸收抑制剂(如依折麦布等)、贝特类药物(如氯贝特、苯扎贝特、非诺贝特等)、PCSK9抑制剂(如Evolocumab和Alirocumab等)、CETP抑制剂(如anacetrapib等)、AMPK激动剂和ACC抑制剂(如GS-0976等)等。The drugs that can be used in combination with the compounds of the present invention can be one or more lipid-lowering drugs, including niacin, statins (such as lovastatin, simvastatin, pravastatin, mevastatin, fluvastatin) Statins, atorvastatin, cerivastatin, rovastatin and pitavastatin), cholesterol absorption inhibitors (such as ezetimibe, etc.), fibrates (such as clofibrate, bezafibrate, fenofibrate) Special class), PCSK9 inhibitors (such as Evolocumab and Alirocumab, etc.), CETP inhibitors (such as anacetrapib, etc.), AMPK agonists and ACC inhibitors (such as GS-0976, etc.).
本发明的式I或式II化合物或其药学上可接受的盐或酯或溶剂化物的用量可根据患者年龄、体重、症状和给药途径等而适当改变。当对成人(约60kg)给药时，式I或式II化合物或其药学上可接受的盐或酯或溶剂化物的给药剂量是1mg～1000mg/次，优选5mg～500mg/次，更优选10mg～60mg/次，每天给药1～3次。也可根据疾病程度的不同和剂型的不同而偏离此剂量范围。The dosage of the compound of formula I or formula II of the present invention or a pharmaceutically acceptable salt or ester or solvate thereof can be appropriately changed according to the patient's age, weight, symptoms and administration route. When administered to an adult (about 60 kg), the dosage of the compound of formula I or formula II or a pharmaceutically acceptable salt or ester or solvate thereof is 1 mg to 1000 mg/time, preferably 5 mg to 500 mg/time, more preferably 10mg～60mg/time, 1 to 3 times a day. It can also deviate from this dosage range according to the degree of disease and the different dosage forms.
有益效果：与现有技术相比，本发明具有如下优点：Beneficial effects: Compared with the prior art, the present invention has the following advantages:
(1)本发明的新型齐墩果酸和δ-齐墩果酸的衍生物具有强效的AMPK激动活性，且其活性显著优于公认的AMPK激动剂AICAR。(1) The novel derivatives of oleanolic acid and delta-oleanolic acid of the present invention have potent AMPK agonist activity, and their activity is significantly better than the recognized AMPK agonist AICAR.
(2)与先导化合物δ-齐墩果酸和齐墩果酸相比，本发明的新型五环三萜类化合物具有更好的口服生物利用度等药代动力学性质。(2) Compared with the lead compounds δ-oleanolic acid and oleanolic acid, the novel pentacyclic triterpenoids of the present invention have better oral bioavailability and other pharmacokinetic properties.
(3)与现有的AMPK激动剂AICAR(因毒副作用大而终止了临床试验)和 MK-8722(可导致不可逆性心肌肥厚副作用)相比，本发明的新型五环三萜类化合物具有非常好的安全性。(3) Compared with the existing AMPK agonist AICAR (clinical trials terminated due to large side effects) and MK-8722 (which can cause irreversible myocardial hypertrophy side effects), the novel pentacyclic triterpenoids of the present invention have very Good security.
(4)本发明的新型五环三萜类化合物作为AMPK激动剂，比现有的AMPK激动剂具有成本低廉、易于制备及潜在的副作用小等优点，其即可单独使用，也可与一种或多种其他类型的预防或治疗AMPK介导的疾病的药物联合使用，有望成为新型预防或治疗AMPK介导的疾病的药物。(4) As AMPK agonists, the novel pentacyclic triterpenoids of the present invention have the advantages of low cost, easy preparation and less potential side effects than existing AMPK agonists. They can be used alone or combined with one Or a variety of other types of drugs for preventing or treating AMPK-mediated diseases are used in combination, which is expected to become a new type of drug for preventing or treating AMPK-mediated diseases.
附图说明Description of the drawings
图1为部分化合物对Huh-7细胞AMPK下游信号通路的影响图(Western Blot检测)。Figure 1 shows the effect of some compounds on the downstream signal pathway of AMPK in Huh-7 cells (Western Blot detection).
具体实施方式Detailed ways
下面通过实施例具体说明本发明的内容。在本发明中，以下所述的实施例是为了更好的阐述本发明，并不是用来限制本发明的范围。在不背离本发明的精神和范围的前提下，可以对本发明进行各种变化和修饰。The content of the present invention will be described in detail through the following examples. In the present invention, the following embodiments are used to better illustrate the present invention, and are not used to limit the scope of the present invention. Various changes and modifications can be made to the present invention without departing from the spirit and scope of the present invention.
实施例1Example 1
(2-(三甲基铵基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺溴化物(化合物A-1)(2-(Trimethylammonium)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide bromide (Compound A-1)
Figure PCTCN2020093248-appb-000036
Figure PCTCN2020093248-appb-000036
取化合物I-1(1.27g，5.0mmol)置于史莱克管中，加入三甲胺的四氢呋喃溶液(20mL，40mmol)，氩气氛围下，55℃加热反应24小时。反应完毕后，用二氯甲烷将反应管中的混合物移至茄形瓶中，减压浓缩，残渣抽滤，滤饼用二氯甲烷(5mL×3)洗涤，得化合物I-2(白色固体，1.166g，产率75％)。Take compound I-1 (1.27 g, 5.0 mmol) and place it in a Shrek tube, add a solution of trimethylamine in tetrahydrofuran (20 mL, 40 mmol), and heat the reaction at 55° C. for 24 hours under an argon atmosphere. After the reaction, the mixture in the reaction tube was transferred to an eggplant-shaped flask with dichloromethane, concentrated under reduced pressure, the residue was suction filtered, and the filter cake was washed with dichloromethane (5mL×3) to obtain compound I-2 (white solid) , 1.166g, yield 75%).
取化合物I-2(1.166g，3.723mmol)悬浮于三氯甲烷和乙醇(20mL，v:v＝7:3)的混合溶液中，冷却至0℃后加入水合肼(340μL)，氩气氛围下，50℃加热回流反应12小时。反应完毕后，反应液抽滤，滤饼用三氯甲烷和乙醇(5mL×3，v:v＝8.5:1.5)的混合溶剂洗涤，滤液浓缩，干燥，得到化合物I-3的粗品直接用于下一步反应。Take compound I-2 (1.166g, 3.723mmol) suspended in a mixed solution of chloroform and ethanol (20mL, v:v=7:3), cool to 0°C, add hydrazine hydrate (340μL), and argon atmosphere The temperature was heated to reflux at 50°C for 12 hours. After the completion of the reaction, the reaction solution was suction filtered, the filter cake was washed with a mixed solvent of chloroform and ethanol (5mL×3, v:v=8.5:1.5), the filtrate was concentrated and dried to obtain the crude product of compound I-3 for direct use Next reaction.
将上步得到化合物I-3的粗品溶于去离子水(2mL)中，冷却至0℃，加入氢溴酸水溶液(2mL，48％)，0℃下搅拌1小时。反应液中加入甲苯和甲醇(5mL，v:v＝4.5:0.5)的混合溶剂，与45℃下减压浓缩，此操作重复两次后大量白色固体析出，抽滤，滤饼用去离子水(1mL×2)洗涤。滤液继续加入甲苯和甲醇(5mL，v:v＝4.5:0.5)的混合溶剂，与45℃下减压浓缩直至除去绝大部分溶剂，加入乙腈(5mL)打浆，抽滤，滤饼用少量甲醇(1mL)洗涤，干燥，得化合物I-4(白色固体，915mg，两步产率93％)： 1H NMR(300MHz,D 2O)δ3.82–3.73(m,2H),3.66–3.57(m,2H),3.30(s,9H). The crude product of compound I-3 obtained in the previous step was dissolved in deionized water (2 mL), cooled to 0°C, aqueous hydrobromic acid (2 mL, 48%) was added, and stirred at 0°C for 1 hour. A mixed solvent of toluene and methanol (5mL, v:v=4.5:0.5) was added to the reaction solution, and concentrated under reduced pressure at 45°C. After this operation was repeated twice, a large amount of white solid precipitated out, filtered with suction, and the filter cake was deionized water (1mL×2) Wash. The filtrate was continuously added with a mixed solvent of toluene and methanol (5mL, v:v=4.5:0.5), and concentrated under reduced pressure at 45°C until most of the solvent was removed. Acetonitrile (5mL) was added to make a slurry, filtered with suction, and a small amount of methanol was used for the filter cake. (1mL) was washed and dried to obtain compound 1-4 (white solid, 915mg, two-step yield 93%): 1 H NMR (300MHz, D 2 O) δ 3.82–3.73 (m, 2H), 3.66–3.57 (m, 2H), 3.30 (s, 9H).
取δ-齐墩果酸(δ-OA，100mg，0.219mmol)溶于N,N-二甲基甲酰胺(8mL)中，加入三乙胺(122μL，0.876mmol)，室温搅拌下加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU,108mg，0.284mmol)，反应1小时后加入化合物I-4(58mg，0.219mmol)，反应液升温至50℃。TLC检测反应完全后，用二氯甲烷(10mL)和去离子水(5mL)处理反应液，水层用二氯甲烷(10mL×2)萃取，合并有机层，去离子水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝10：1)纯化，得化合物I-5(白色固体，41mg，产率30％)。Dissolve δ-oleanolic acid (δ-OA, 100mg, 0.219mmol) in N,N-dimethylformamide (8mL), add triethylamine (122μL, 0.876mmol), add 2- with stirring at room temperature (7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU, 108mg, 0.284mmol), after reacting for 1 hour, compound I-4 (58mg, 0.219mmol), the reaction solution was heated to 50°C. After the completion of the reaction detected by TLC, the reaction solution was treated with dichloromethane (10mL) and deionized water (5mL), the aqueous layer was extracted with dichloromethane (10mL×2), the organic layers were combined and washed with deionized water (5mL×2) , Dried over anhydrous sodium sulfate, filtered, concentrated the filtrate, and purified by silica gel column chromatography (dichloromethane: methanol = 10:1) to obtain compound I-5 (white solid, 41 mg, yield 30%).
取化合物I-5(175mg，0.281mmol)悬浮于无水二氯甲烷(10mL)中，依次加入N,N'-二环己基碳酰亚胺(DCC,232mg，1.124mmol)、4-二甲氨基吡啶(DMAP,34mg，0.281mmol)和邻苯二甲酸单苄酯(86mg，0.337mmol)，室温搅拌反应。TLC检测反应完全后，抽滤，滤液用去离子水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝30：1)纯化，得化合物I-6(白色固体，100mg，产率41％)。Suspend compound I-5 (175mg, 0.281mmol) in anhydrous dichloromethane (10mL), add N,N'-dicyclohexylcarbimide (DCC, 232mg, 1.124mmol), 4-dimethyl Aminopyridine (DMAP, 34 mg, 0.281 mmol) and monobenzyl phthalate (86 mg, 0.337 mmol) were reacted with stirring at room temperature. After the completion of the reaction detected by TLC, it was filtered with suction, the filtrate was washed with deionized water (5mL×2), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 30:1). Compound I-6 (white solid, 100 mg, yield 41%) was obtained.
取化合物I-6(100mg，0.116mmol)溶于四氢呋喃(8mL)中，加入10％钯碳(10mg)，氢气氛围下室温搅拌反应过夜。TLC检测反应完全后，过滤硅藻土，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝10：1)纯化，得化合物A-1(白色固体，66mg，产率73％)： 1H NMR(300MHz,DMSO-d 6)δ13.18(s,1H),7.79–7.69(m,1H),7.69–7.57(m,3H),7.17–7.08(m,1H),4.74–4.60(m,1H),3.59–3.43(m,2H),3.12(s,9H),2.81(d,J＝14.1Hz,1H),2.49–2.37(m,1H),2.17(d,J＝13.2Hz,1H),1.18(s,3H),0.93(s,3H),0.91(s,9H),0.84(s,3H),0.74(s,3H). ESI-MS:m/z 689.6[M-Br] +。 The compound I-6 (100 mg, 0.116 mmol) was dissolved in tetrahydrofuran (8 mL), 10% palladium on carbon (10 mg) was added, and the reaction was stirred overnight at room temperature under a hydrogen atmosphere. After the completion of the reaction detected by TLC, Celite was filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 10:1) to obtain compound A-1 (white solid, 66 mg, yield 73%): 1 H NMR (300MHz, DMSO-d 6 ) δ13.18(s,1H), 7.79-7.69(m,1H), 7.69-7.57(m,3H), 7.17-7.08(m,1H), 4.74-4.60( m,1H),3.59–3.43(m,2H), 3.12(s,9H), 2.81(d,J=14.1Hz,1H), 2.49–2.37(m,1H), 2.17(d,J=13.2Hz ,1H),1.18(s,3H),0.93(s,3H),0.91(s,9H),0.84(s,3H),0.74(s,3H). ESI-MS: m/z 689.6(M- Br] + .
实施例2Example 2
N-(2-(二甲基氨基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-3)N-(2-(Dimethylamino)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide (Compound A-3)
Figure PCTCN2020093248-appb-000037
Figure PCTCN2020093248-appb-000037
取δ-齐墩果酸(5g，0.011mol)溶于无水吡啶(35mL)中，搅拌下缓慢滴加醋酸酐(4mL)，滴加完毕后加热回流反应1小时。TLC检测反应完全后，反应液冷却至室温，将其滴入冰水(20mL×2)中，析出大量白色固体，抽滤，滤饼用水洗(10mL×3)，正己烷洗(10mL×3)，真空干燥，乙醇重结晶，得化合物II-1(白色固体，4.52g，产率83％)。Take δ-oleanolic acid (5g, 0.011mol) and dissolve it in anhydrous pyridine (35mL), slowly add acetic anhydride (4mL) dropwise with stirring, and heat and reflux for 1 hour after the addition is complete. After the completion of the reaction detected by TLC, the reaction solution was cooled to room temperature and dropped into ice water (20mL×2). A large amount of white solid precipitated out. The filter cake was washed with water (10mL×3) and n-hexane (10mL×3). ), vacuum drying, and ethanol recrystallization to obtain compound II-1 (white solid, 4.52 g, yield 83%).
取化合物II-1(150mg，0.30mmol)溶于无水二氯甲烷(5mL)中，搅拌下缓慢滴加草酰氯(130μL，1.50mmol)和N,N-二甲基甲酰胺(1滴)，室温反应3小时。TLC检测反应完全后，减压蒸除溶剂，得化合物II-2(黄色固体，155mg，产率100％)。Dissolve compound II-1 (150mg, 0.30mmol) in anhydrous dichloromethane (5mL), slowly add oxalyl chloride (130μL, 1.50mmol) and N,N-dimethylformamide (1 drop) while stirring. , React at room temperature for 3 hours. After the completion of the reaction detected by TLC, the solvent was evaporated under reduced pressure to obtain compound II-2 (yellow solid, 155 mg, yield 100%).
将N,N-二甲基乙二胺(66μL，0.60mmol)溶于无水二氯甲烷(5mL)中，搅拌下缓慢滴加化合物II-2(155mg，0.30mmol)的无水二氯甲烷(5mL)溶液和三乙胺(84μL，0.60mmol)，室温反应5小时。TLC检测反应完全后，加入二氯甲烷(10mL)稀释反应液，反应液依次用饱和碳酸氢钠溶液(10mL×3)、水(10mL×3)和饱和食盐水(10mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物II-3(白色固体，143mg，产率84％)。N,N-Dimethylethylenediamine (66μL, 0.60mmol) was dissolved in dry dichloromethane (5mL), and compound II-2 (155mg, 0.30mmol) in dry dichloromethane was slowly added dropwise with stirring (5mL) The solution and triethylamine (84μL, 0.60mmol) were reacted at room temperature for 5 hours. After the completion of the reaction detected by TLC, dichloromethane (10mL) was added to dilute the reaction solution, and the reaction solution was washed with saturated sodium bicarbonate solution (10mL×3), water (10mL×3) and saturated brine (10mL×3) successively. Drying with sodium sulfate, filtering, concentrating the filtrate, and purifying by silica gel column chromatography (dichloromethane:methanol=20:1) to obtain compound II-3 (white solid, 143 mg, yield 84%).
将化合物II-3(141mg，0.248mmol)悬浮于甲醇(10mL)中，加入氢氧化钾(140mg，2.48mmol)，室温搅拌过夜。TLC检测反应完全后，反应液冷却 至室温，减压蒸除溶剂，残余物用乙酸乙酯(20mL)和水(20mL)处理，有机相依次用水(20mL×3)和饱和食盐水(20mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物II-4(白色固体，87mg，产率67％)。Compound II-3 (141 mg, 0.248 mmol) was suspended in methanol (10 mL), potassium hydroxide (140 mg, 2.48 mmol) was added, and the mixture was stirred at room temperature overnight. After the completion of the reaction detected by TLC, the reaction solution was cooled to room temperature, and the solvent was evaporated under reduced pressure. The residue was treated with ethyl acetate (20mL) and water (20mL). The organic phase was sequentially water (20mL×3) and saturated brine (20mL× 3) Wash, dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and purify by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound II-4 (white solid, 87 mg, yield 67%).
将化合物II-4(87mg，0.165mmol)溶于无水吡啶(10mL)中，加入邻苯二甲酸酐(245mg，1.65mmol)和4-二甲氨基吡啶(20mg，0.165mmol)，反应液升温至115℃搅拌过夜。TLC检测反应完全后，加入二氯甲烷(20mL)稀释反应液，反应液依次用1N稀盐酸(10mL×3)和水(10mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物A-3(白色固体，93mg，产率84％)： 1H NMR(300MHz,CDCl 3)δ7.84–7.63(m,1H),7.63–7.51(m,1H),7.51–7.30(m,2H),6.72–6.50(m,1H),4.82–4.59(m,1H),3.77–3.43(m,2H),3.07–2.83(m,2H),2.83–2.70(m,1H),2.63(s,6H),2.41(d,J＝13.7Hz,1H),2.36–2.18(m,1H),1.16(s,3H),0.98(s,3H),0.87(s,9H),0.83(s,3H),0.72(s,3H).ESI-MS:m/z 673.3[M-H] -。 Compound II-4 (87mg, 0.165mmol) was dissolved in anhydrous pyridine (10mL), phthalic anhydride (245mg, 1.65mmol) and 4-dimethylaminopyridine (20mg, 0.165mmol) were added, and the reaction solution was heated Stir at 115°C overnight. After the completion of the reaction detected by TLC, dichloromethane (20mL) was added to dilute the reaction solution. The reaction solution was washed with 1N dilute hydrochloric acid (10mL×3) and water (10mL×3) successively, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound A-3 (white solid, 93 mg, yield 84%): 1 H NMR (300MHz, CDCl 3 ) δ 7.84–7.63 (m ,1H), 7.63--7.51(m,1H), 7.51--7.30(m,2H), 6.72--6.50(m,1H), 4.82--4.59(m,1H), 3.77--3.43(m,2H), 3.07 –2.83(m,2H),2.83–2.70(m,1H),2.63(s,6H),2.41(d,J=13.7Hz,1H),2.36–2.18(m,1H),1.16(s,3H) ), 0.98 (s, 3H), 0.87 (s, 9H), 0.83 (s, 3H), 0.72 (s, 3H). ESI-MS: m/z 673.3 [MH] - .
实施例3Example 3
N-(2-(1-哌啶基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-4)N-(2-(1-piperidinyl)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide (compound A-4)
Figure PCTCN2020093248-appb-000038
Figure PCTCN2020093248-appb-000038
参照实施例2的方法，将N,N-二甲基乙二胺替换成1-(2-氨基乙基)哌啶，制得化合物A-4： 1H NMR(300MHz,CDCl 3)δ7.74(d,J＝7.1Hz,1H),7.52(d,J＝6.9Hz,1H),7.47–7.29(m,2H),6.77–6.64(m,1H),4.78–4.59(m,1H),3.75–3.46(m,2H),3.08–2.67(m,7H),2.43(d,J＝13.6Hz,1H),2.31(d,J＝10.1Hz,1H),1.16(s,3H),0.98(s,3H),0.88(s,3H),0.86(s,6H),0.79(s,3H),0.72(s,3H).ESI-MS:m/z 715.5[M+H] +。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with 1-(2-aminoethyl)piperidine to obtain compound A-4: 1 H NMR (300MHz, CDCl 3 )δ7. 74(d,J=7.1Hz,1H),7.52(d,J=6.9Hz,1H),7.47–7.29(m,2H),6.77–6.64(m,1H),4.78–4.59(m,1H) ,3.75–3.46(m,2H),3.08–2.67(m,7H),2.43(d,J=13.6Hz,1H), 2.31(d,J=10.1Hz,1H),1.16(s,3H), 0.98 (s, 3H), 0.88 (s, 3H), 0.86 (s, 6H), 0.79 (s, 3H), 0.72 (s, 3H). ESI-MS: m/z 715.5 [M+H] + .
实施例4Example 4
N-(2-(4-吗啉基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-5)N-(2-(4-morpholinyl)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide (compound A-5)
Figure PCTCN2020093248-appb-000039
Figure PCTCN2020093248-appb-000039
参照实施例2的方法，将N,N-二甲基乙二胺替换成4-(2-氨基乙基)吗啉，制得化合物A-5： 1H NMR(500MHz,CDCl 3)δ7.76(d,J＝5.8Hz,1H),7.63(d,J＝5.8Hz,1H),7.52–7.41(m,2H),6.57–6.46(m,1H),4.76–4.67(m,1H),3.76(s,4H),3.57–3.38(m,2H),2.81(d,J＝13.8Hz,1H),2.63(s,6H),2.45(d,J＝13.5Hz,1H),2.42–2.33(m,1H),1.18(s,3H),0.95(s,3H),0.90(s,3H),0.89(s,3H),0.87(s,3H),0.84(s,3H),0.74(s,3H).ESI-MS:m/z 715.5[M-H] -。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with 4-(2-aminoethyl)morpholine to obtain compound A-5: 1 H NMR (500MHz, CDCl 3 )δ7. 76(d,J=5.8Hz,1H), 7.63(d,J=5.8Hz,1H), 7.52–7.41(m,2H), 6.57–6.46(m,1H), 4.76–4.67(m,1H) ,3.76(s,4H),3.57–3.38(m,2H), 2.81(d,J=13.8Hz,1H), 2.63(s,6H), 2.45(d,J=13.5Hz,1H),2.42– 2.33(m,1H),1.18(s,3H),0.95(s,3H),0.90(s,3H),0.89(s,3H),0.87(s,3H),0.84(s,3H),0.74 (s, 3H). ESI-MS: m/z 715.5 [MH] - .
实施例5Example 5
N-(2-(1-四氢吡咯基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-6)N-(2-(1-Tetrahydropyrrolyl)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide (Compound A-6 )
Figure PCTCN2020093248-appb-000040
Figure PCTCN2020093248-appb-000040
参照实施例2的方法，将N,N-二甲基乙二胺替换成1-(2-氨基乙基)吡咯烷，制得化合物A-6： 1H NMR(300MHz,CDCl 3)δ7.73(d,J＝7.3Hz,1H),7.49(d,J＝7.1Hz,1H),7.37(dt,J＝19.0,7.3Hz,2H),4.78–4.65(m,1H),3.72–3.54(m,2H),3.26–3.08(m,4H),3.08–2.95(m,2H),2.77(d,J＝13.3Hz,1H),2.42(d,J＝13.5Hz,1H),2.31(d,J＝12.0Hz,1H),1.16(s,3H),1.02(s,3H),0.88(s,3H),0.86(s,6H),0.82(s,3H),0.72(s,3H).ESI-MS:m/z 699.3[M-H] -。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with 1-(2-aminoethyl)pyrrolidine to obtain compound A-6: 1 H NMR (300MHz, CDCl 3 )δ7. 73(d,J=7.3Hz,1H),7.49(d,J=7.1Hz,1H),7.37(dt,J=19.0,7.3Hz,2H),4.78–4.65(m,1H),3.72–3.54 (m, 2H), 3.26–3.08 (m, 4H), 3.08–2.95 (m, 2H), 2.77 (d, J = 13.3 Hz, 1H), 2.42 (d, J = 13.5 Hz, 1H), 2.31 ( d,J=12.0Hz,1H),1.16(s,3H),1.02(s,3H),0.88(s,3H),0.86(s,6H),0.82(s,3H),0.72(s,3H) ).ESI-MS: m/z 699.3[MH] - .
实施例6Example 6
N-(2-(4-甲基-1-哌嗪基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-8)N-(2-(4-Methyl-1-piperazinyl)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide( Compound A-8)
Figure PCTCN2020093248-appb-000041
Figure PCTCN2020093248-appb-000041
参照实施例2的方法，将N,N-二甲基乙二胺替换成1-(2-氨基乙基)-4-甲基哌嗪，制得化合物A-8： 1H NMR(300MHz,CDCl 3)δ7.75(d,J＝6.9Hz,1H),7.64(d,J＝7.0Hz,1H),7.53–7.35(m,2H),6.46–6.36(m,1H),4.80–4.69(m,1H),3.54–3.30(m,2H),3.04–2.80(m,4H),2.73(s,5H),2.57(s,5H),2.53–2.45(m,1H),2.46–2.38(m,1H),1.24(s,3H),1.00(s,3H),0.97(s,3H),0.93(s,9H),0.78(s,3H).ESI-MS:m/z 730.5[M+H] +。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with 1-(2-aminoethyl)-4-methylpiperazine to obtain compound A-8: 1 H NMR (300MHz, CDCl 3 )δ7.75(d,J=6.9Hz,1H), 7.64(d,J=7.0Hz,1H), 7.53–7.35(m,2H), 6.46–6.36(m,1H), 4.80–4.69 (m,1H),3.54–3.30(m,2H),3.04–2.80(m,4H), 2.73(s,5H), 2.57(s,5H), 2.53–2.45(m,1H), 2.46–2.38 (m,1H),1.24(s,3H),1.00(s,3H),0.97(s,3H),0.93(s,9H),0.78(s,3H).ESI-MS:m/z 730.5[ M+H] + .
实施例7Example 7
N-(2-(4-吗啉基)乙基)-3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-12)N-(2-(4-morpholinyl)ethyl)-3β-(2-(2-acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-13(18)-ene-28 -Amide (Compound A-12)
Figure PCTCN2020093248-appb-000042
Figure PCTCN2020093248-appb-000042
取齐墩果酸(OA，20g,0.044mol)溶于N,N-二甲基甲酰胺(200mL)中，加入碳酸钾(6g,0.044mol)，搅拌下缓慢滴加溴化苄(8mL，0.044mol)，滴加完毕后将反应液升温至85℃，反应10小时。TLC检测反应完全后，趁热抽滤，滤饼用热的N,N-二甲基甲酰胺(10mL×3)洗涤，滤液冷却至室温后滴入冰水(100mL×2)中，析出大量白色固体，抽滤，滤饼用水洗(50mL×3)，正己烷洗(50mL×3)，真空干燥，得化合物III-1(白色固体，22.02g，产率92％)。Dissolve oleanolic acid (OA, 20g, 0.044mol) in N,N-dimethylformamide (200mL), add potassium carbonate (6g, 0.044mol), slowly add benzyl bromide (8mL, 0.044 mol). After the dropping, the reaction solution was heated to 85°C and reacted for 10 hours. After the completion of the reaction detected by TLC, it was filtered while hot, the filter cake was washed with hot N,N-dimethylformamide (10mL×3), the filtrate was cooled to room temperature and then dropped into ice water (100mL×2), a large amount of precipitation The white solid was filtered with suction, the filter cake was washed with water (50 mL×3), washed with n-hexane (50 mL×3), and dried under vacuum to obtain compound III-1 (white solid, 22.02 g, yield 92%).
取化合物III-1(22g,0.040mol)溶于二氯甲烷(200mL)中，加入质子化蒙脱土(44g)，将反应液升温至40℃，反应8小时。TLC检测反应完全后，冷却至室温，抽滤，滤饼用二氯甲烷(50mL×3)洗涤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝10：1)纯化，得化合物III-2(白色固体，16.52g，产率75％)。The compound III-1 (22g, 0.040mol) was dissolved in dichloromethane (200mL), protonated montmorillonite (44g) was added, the reaction solution was heated to 40°C and reacted for 8 hours. After the completion of the reaction detected by TLC, it was cooled to room temperature, filtered with suction, the filter cake was washed with dichloromethane (50 mL×3), the filtrate was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain the compound III-2 (white solid, 16.52 g, yield 75%).
将化合物III-2(1g，1.829mmol)溶于无水吡啶(10mL)中，加入邻苯二甲酸酐(2.708g，18.29mmol)和4-二甲氨基吡啶(223mg，1.829mmol)，反应液升温至115℃搅拌过夜。TLC检测反应完全后，加入二氯甲烷(30mL)稀释反应液，反应液依次用1N稀盐酸(10mL×3)和水(10mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物III-3(白色固体，925mg，产率73％)。Compound III-2 (1g, 1.829mmol) was dissolved in dry pyridine (10mL), phthalic anhydride (2.708g, 18.29mmol) and 4-dimethylaminopyridine (223mg, 1.829mmol) were added to the reaction solution The temperature was raised to 115°C and stirred overnight. After the completion of the reaction was detected by TLC, dichloromethane (30mL) was added to dilute the reaction solution. The reaction solution was washed with 1N diluted hydrochloric acid (10mL×3) and water (10mL×3) successively, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound III-3 (white solid, 925 mg, yield 73%).
取化合物III-3(150mg，0.216mmol)溶于无水二氯甲烷(6mL)中，依次加入N,N'-二环己基碳酰亚胺(134mg，0.647mmol)、4-二甲氨基吡啶(27mg， 0.216mmol)和N-乙酰乙醇胺(30μL，0.324mmol)，室温搅拌反应。TLC检测反应完全后，抽滤，滤液用水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝3：1)纯化，得化合物III-4(白色固体，159mg，产率95％)。Take compound III-3 (150mg, 0.216mmol) and dissolve it in dry dichloromethane (6mL), add N,N'-dicyclohexylcarbimide (134mg, 0.647mmol), 4-dimethylaminopyridine in turn (27mg, 0.216mmol) and N-acetylethanolamine (30μL, 0.324mmol), the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, it was suction filtered, the filtrate was washed with water (5mL×2), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate=3:1) to obtain the compound III-4 (white solid, 159 mg, yield 95%).
取化合物III-4(159mg，0.204mmol)溶于四氢呋喃(10mL)中，加入10％钯碳(16mg)，氢气氛围下室温搅拌反应过夜。TLC检测反应完全后，过滤硅藻土，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝1：1)纯化，得化合物III-5(白色固体，90mg，产率64％)The compound III-4 (159 mg, 0.204 mmol) was dissolved in tetrahydrofuran (10 mL), 10% palladium on carbon (16 mg) was added, and the reaction was stirred at room temperature under a hydrogen atmosphere overnight. After the completion of the reaction detected by TLC, celite was filtered, the filtrate was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound III-5 (white solid, 90 mg, yield 64%)
取化合物III-5(100mg，0.145mmol)溶于N,N-二甲基甲酰胺(6mL)中，加入三乙胺(40μL，0.290mmol)，室温搅拌下加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(72mg，0.188mmol)，反应1小时后加入N-(2-氨基乙基)吗啉(23μL，0.174mmol)，反应液升温至50℃。TLC检测反应完全后，用二氯甲烷(10mL)和水(5mL)处理反应液，水层用二氯甲烷(10mL×2)萃取，合并有机层，水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝30：1)纯化，得化合物A-12(白色固体，80mg，产率69％)： 1H NMR(300MHz,CDCl 3)δ7.81(d,J＝7.3Hz,1H),7.66(d,J＝7.0Hz,1H),7.63–7.50(m,2H),6.60–6.49(m,1H),6.49–6.36(m,1H),4.77–4.65(m,1H),4.44(t,J＝4.8Hz,2H),3.76–3.56(m,6H),3.47–3.25(m,2H),2.86(d,J＝15.4Hz,1H),2.53–2.33(m,8H),2.03(s,3H),1.21(s,3H),0.97(s,3H),0.96(s,6H),0.94(s,3H),0.89(s,3H),0.75(s,3H).ESI-MS:m/z 824.6[M+Na] +。 Dissolve compound III-5 (100mg, 0.145mmol) in N,N-dimethylformamide (6mL), add triethylamine (40μL, 0.290mmol), add 2-(7-oxybenzoic acid) under stirring at room temperature Triazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (72mg, 0.188mmol), after 1 hour of reaction, add N-(2-aminoethyl)morpholine (23μL, 0.174 mmol), the reaction solution was heated to 50°C. After the completion of the reaction detected by TLC, the reaction solution was treated with dichloromethane (10mL) and water (5mL), the aqueous layer was extracted with dichloromethane (10mL×2), the organic layers were combined, washed with water (5mL×2), and anhydrous sulfuric acid Dry sodium, filter, concentrate the filtrate, and purify by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain compound A-12 (white solid, 80 mg, yield 69%): 1 H NMR (300MHz, CDCl 3 )δ7.81(d,J=7.3Hz,1H), 7.66(d,J=7.0Hz,1H), 7.63–7.50(m,2H), 6.60–6.49(m,1H), 6.49–6.36( m,1H),4.77–4.65(m,1H), 4.44(t,J=4.8Hz,2H), 3.76–3.56(m,6H), 3.47–3.25(m,2H), 2.86(d,J= 15.4Hz, 1H), 2.53-2.33 (m, 8H), 2.03 (s, 3H), 1.21 (s, 3H), 0.97 (s, 3H), 0.96 (s, 6H), 0.94 (s, 3H), 0.89 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 824.6 [M+Na] + .
实施例8Example 8
N-(2-(4-甲基-1-哌嗪基)乙基)-3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-14)N-(2-(4-Methyl-1-piperazinyl)ethyl)-3β-(2-(2-acetamidoethoxyyl)benzoyl)oxy-oleanorane-13(18 )-Ene-28-amide (Compound A-14)
Figure PCTCN2020093248-appb-000043
Figure PCTCN2020093248-appb-000043
参照实施例7的方法，将4-(2-氨基乙基)吗啉替换成1-(2-氨基乙基)-4-甲基哌嗪，制得化合物A-14： 1H NMR(300MHz,CDCl 3)δ7.81(d,J＝7.2Hz,1H),7.66(d,J＝7.0Hz,1H),7.62–7.48(m,2H),6.58–6.48(m,1H),6.45–6.36(m,1H),4.76–4.67(m,1H),4.44(t,J＝4.8Hz,2H),3.73–3.55(m,2H),3.43–3.21(m,2H),2.85(d,J＝15.3Hz,1H),2.71–2.51(m,6H),2.51–2.42(m,4H),2.42–2.38(m, 1H),2.36(s,4H),2.03(s,3H),1.20(s,3H),0.97(s,3H),0.95(s,9H),0.90(s,3H),0.75(s,3H).ESI-MS:m/z 815.7[M+H] +。 Referring to the method of Example 7, the 4-(2-aminoethyl)morpholine was replaced with 1-(2-aminoethyl)-4-methylpiperazine to obtain compound A-14: 1 H NMR (300MHz ,CDCl 3 )δ7.81(d,J=7.2Hz,1H),7.66(d,J=7.0Hz,1H),7.62–7.48(m,2H),6.58–6.48(m,1H),6.45– 6.36(m,1H),4.76–4.67(m,1H), 4.44(t,J=4.8Hz,2H), 3.73–3.55(m,2H), 3.43–3.21(m,2H), 2.85(d, J = 15.3Hz, 1H), 2.71–2.51 (m, 6H), 2.51–2.42 (m, 4H), 2.42–2.38 (m, 1H), 2.36 (s, 4H), 2.03 (s, 3H), 1.20 (s, 3H), 0.97 (s, 3H), 0.95 (s, 9H), 0.90 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 815.7 [M+H] + .
实施例9Example 9
N-(2-(三甲基铵基)乙基)-3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺溴化物(化合物A-15)N-(2-(Trimethylammonium)ethyl)-3β-(2-(2-Acetylaminoethoxyyl)benzoyl)oxy-oleanorane-13(18)-ene-28 -Amide bromide (Compound A-15)
Figure PCTCN2020093248-appb-000044
Figure PCTCN2020093248-appb-000044
参照实施例7的方法制得化合物III-5，取化合物III-5(100mg，0.145mmol)溶于N,N-二甲基甲酰胺(8mL)中，加入三乙胺(80μL，0.580mmol)，室温搅拌下加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(72mg，0.188mmol)，反应1小时后加入化合物I-4(60mg，0.174mmol)(由实施例1制得)，反应液升温至50℃。TLC检测反应完全后，用二氯甲烷(10mL)和去离子水(5mL)处理反应液，水层用二氯甲烷(10mL×2)萃取，合并有机层，去离子水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝30：1)纯化，得化合物A-15(白色固体，80mg，产率65％)： 1H NMR(300MHz,DMSO-d 6)δ8.09–7.98(m,1H),7.83–7.74(m,2H),7.74–7.64(m,2H),7.17–7.06(m,1H),4.74–4.61(m,1H),4.30–4.14(m,2H),3.59–3.45(m,2H),3.44–3.33(m,4H),3.13(s,9H),2.91–2.72(m,1H),2.48–2.39(m,1H),2.18(d,J＝12.1Hz,1H),1.84(s,3H),1.19(s,3H),0.92(s,12H),0.86(s,3H),0.75(s,3H).ESI-MS:m/z 774.6[M-Br] +。 The compound III-5 was prepared according to the method of Example 7. The compound III-5 (100mg, 0.145mmol) was dissolved in N,N-dimethylformamide (8mL), and triethylamine (80μL, 0.580mmol) was added. , Add 2-(7-oxybenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (72mg, 0.188mmol) under stirring at room temperature, and add compound I after reacting for 1 hour -4 (60mg, 0.174mmol) (prepared in Example 1), the reaction solution was heated to 50°C. After the completion of the reaction detected by TLC, the reaction solution was treated with dichloromethane (10mL) and deionized water (5mL), the aqueous layer was extracted with dichloromethane (10mL×2), the organic layers were combined and washed with deionized water (5mL×2) , Dried over anhydrous sodium sulfate, filtered, concentrated the filtrate, and purified by silica gel column chromatography (dichloromethane:methanol=30:1) to obtain compound A-15 (white solid, 80mg, yield 65%): 1 H NMR (300MHz, DMSO-d 6 )δ8.09-7.98(m,1H), 7.83-7.74(m,2H), 7.74-7.64(m,2H), 7.17-7.06(m,1H), 4.74-4.61( m,1H), 4.30--4.14(m,2H), 3.59--3.45(m,2H), 3.44--3.33(m,4H), 3.13(s,9H), 2.91--2.72(m,1H), 2.48-- 2.39(m,1H), 2.18(d,J=12.1Hz,1H),1.84(s,3H),1.19(s,3H),0.92(s,12H),0.86(s,3H),0.75(s , 3H). ESI-MS: m/z 774.6 [M-Br] + .
实施例10Example 10
N-(3-(三甲基铵基)丙基)-3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺溴化物(化合物A-16)N-(3-(Trimethylammonium)propyl)-3β-(2-(2-acetamidoethoxyyl)benzoyl)oxy-oleanorane-13(18)-ene-28 -Amide bromide (Compound A-16)
Figure PCTCN2020093248-appb-000045
Figure PCTCN2020093248-appb-000045
Figure PCTCN2020093248-appb-000046
Figure PCTCN2020093248-appb-000046
参照实施例1的方法，将化合物I-1替换成I-1’，制得化合物I-4’。再参照实施例9的方法，将化合物I-4替换成I-4’，制得化合物A-16： 1H NMR(300MHz,DMSO-d 6)δ8.08–7.96(m,1H),7.79–7.71(m,2H),7.67(d,J＝8.6Hz,2H),7.04–6.88(m,1H),4.71–4.60(m,1H),4.29–4.13(m,2H),3.41–3.29(m,2H),3.29–3.18(m,2H),3.18–3.08(m,2H),3.04(s,9H),2.79(d,J＝12.5Hz,1H),2.43(d,J＝14.3Hz,1H),2.17(d,J＝12.9Hz,1H),1.81(s,3H),1.16(s,3H),0.90(s,12H),0.83(s,3H),0.72(s,3H).ESI-MS:m/z 788.6[M-Br] +。 Referring to the method of Example 1, the compound I-1 was replaced with I-1' to prepare the compound I-4'. Again referring to the method of Example 9, the compound I-4 was replaced with I-4' to obtain the compound A-16: 1 H NMR (300MHz, DMSO-d 6 ) δ 8.08–7.96 (m, 1H), 7.79 –7.71(m,2H), 7.67(d,J=8.6Hz,2H), 7.04–6.88(m,1H), 4.71–4.60(m,1H), 4.29–4.13(m,2H), 3.41–3.29 (m,2H),3.29–3.18(m,2H),3.18–3.08(m,2H),3.04(s,9H),2.79(d,J=12.5Hz,1H),2.43(d,J=14.3 Hz, 1H), 2.17 (d, J = 12.9 Hz, 1H), 1.81 (s, 3H), 1.16 (s, 3H), 0.90 (s, 12H), 0.83 (s, 3H), 0.72 (s, 3H) ). ESI-MS: m/z 788.6[M-Br] + .
实施例11Example 11
N-(2-(三甲基铵基)乙基)-3β-(2-甲氧酰基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺溴化物(化合物A-17)N-(2-(Trimethylammonium)ethyl)-3β-(2-methoxyacylbenzoyl)oxy-oleanorane-13(18)-ene-28-amide bromide (compound A-17)
Figure PCTCN2020093248-appb-000047
Figure PCTCN2020093248-appb-000047
参照实施例7的方法制得化合物III-2，取化合物III-2(150mg，0.274mmol)溶于无水二氯甲烷(6mL)中，依次加入N,N'-二环己基碳酰亚胺(170mg，0.823mmol)、4-二甲氨基吡啶(34mg，0.274mmol)和邻苯二甲酸单甲酯(54mg， 0.302mmol)，室温搅拌反应。TLC检测反应完全后，抽滤，滤液用水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝20：1)纯化，得化合物IV-1(白色固体，159mg，产率82％)。The compound III-2 was prepared according to the method of Example 7. The compound III-2 (150 mg, 0.274 mmol) was dissolved in anhydrous dichloromethane (6 mL), and N,N'-dicyclohexylcarbimide was added in sequence (170 mg, 0.823 mmol), 4-dimethylaminopyridine (34 mg, 0.274 mmol) and monomethyl phthalate (54 mg, 0.302 mmol), the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, filter with suction, wash the filtrate with water (5mL×2), dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and purify by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain the compound IV-1 (white solid, 159 mg, yield 82%).
取化合物IV-1(159mg，0.224mmol)溶于四氢呋喃(10mL)中，加入10％钯碳(20mg)，氢气氛围下室温搅拌过夜。TLC检测反应完全后，过滤硅藻土，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝1：1)纯化，得化合物IV-2(白色固体，118mg，产率85％)。The compound IV-1 (159 mg, 0.224 mmol) was dissolved in tetrahydrofuran (10 mL), 10% palladium on carbon (20 mg) was added, and the mixture was stirred overnight at room temperature under a hydrogen atmosphere. After the completion of the reaction was detected by TLC, Celite was filtered, the filtrate was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound IV-2 (white solid, 118 mg, yield 85%).
取化合物IV-2(150mg，0.242mmol)溶于N,N-二甲基甲酰胺(8mL)中，加入三乙胺(135μL，0.970mmol)，室温搅拌下加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(120mg，0.315mmol)，反应1小时后加入化合物I-4(128mg，0.485mmol)，反应液升温至50℃。TLC检测反应完全后，用二氯甲烷(10mL)和去离子水(5mL)处理反应液，水层用二氯甲烷(10mL×2)萃取，合并有机层，去离子水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物A-17(白色固体，77mg，产率40％)： 1H NMR(300MHz,DMSO-d 6)δ7.82–7.64(m,4H),7.18–7.06(m,1H),4.74–4.64(m,1H),3.83(s,3H),3.59–3.45(m,2H),3.39–3.34(m,2H),3.13(s,9H),2.83(d,J＝13.4Hz,1H),2.50–2.41(m,1H),2.24–2.11(m,1H),1.19(s,3H),0.93(s,6H),0.92(s,6H),0.86(s,3H),0.75(s,3H).ESI-MS:m/z 703.6[M-Br] +。 Take compound IV-2 (150mg, 0.242mmol) dissolved in N,N-dimethylformamide (8mL), add triethylamine (135μL, 0.970mmol), add 2-(7-oxybenzoic acid) under stirring at room temperature Triazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (120mg, 0.315mmol), after 1 hour of reaction, compound I-4 (128mg, 0.485mmol) was added, and the reaction solution was heated to 50°C. After the completion of the reaction detected by TLC, the reaction solution was treated with dichloromethane (10mL) and deionized water (5mL), the aqueous layer was extracted with dichloromethane (10mL×2), the organic layers were combined and washed with deionized water (5mL×2) , Dried over anhydrous sodium sulfate, filtered, concentrated the filtrate, and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound A-17 (white solid, 77 mg, yield 40%): 1 H NMR (300MHz,DMSO-d 6 )δ7.82-7.64(m,4H), 7.18-7.06(m,1H), 4.74-4.64(m,1H), 3.83(s,3H), 3.59-3.45(m, 2H), 3.39–3.34(m,2H), 3.13(s,9H), 2.83(d,J=13.4Hz,1H), 2.50–2.41(m,1H), 2.24–2.11(m,1H), 1.19 (s, 3H), 0.93 (s, 6H), 0.92 (s, 6H), 0.86 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 703.6 [M-Br] + .
实施例12Example 12
N-(3-(三甲基铵基)丙基)-3β-(2-甲氧酰基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺溴化物(化合物A-18)N-(3-(Trimethylammonium)propyl)-3β-(2-Methoxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide bromide (compound A-18)
Figure PCTCN2020093248-appb-000048
Figure PCTCN2020093248-appb-000048
参照实施例11的方法，将化合物I-4替换成I-4’，制得化合物A-18： 1H NMR(300MHz,DMSO-d 6)δ7.80–7.63(m,4H),7.02–6.89(m,1H),4.76–4.62(m,1H),3.82(s,3H),3.30–3.20(m,2H),3.20–3.10(m,2H),3.06(s,9H),2.81(d,J＝14.7Hz,1H),2.46(d,J＝14.8Hz,1H),2.20(d,J＝13.1Hz,1H),1.19(s,3H),0.93(s,12H),0.86(s,3H),0.74(s,3H).ESI-MS:m/z 717.7[M-Br] +。 According to the method of Example 11, compound I-4 was replaced with I-4' to obtain compound A-18: 1 H NMR (300MHz, DMSO-d 6 ) δ 7.80–7.63 (m, 4H), 7.02– 6.89 (m, 1H), 4.76 - 4.62 (m, 1H), 3.82 (s, 3H), 3.30 - 3.20 (m, 2H), 3.20 - 3.10 (m, 2H), 3.06 (s, 9H), 2.81 ( d, J = 14.7 Hz, 1H), 2.46 (d, J = 14.8 Hz, 1H), 2.20 (d, J = 13.1 Hz, 1H), 1.19 (s, 3H), 0.93 (s, 12H), 0.86 ( s, 3H), 0.74 (s, 3H). ESI-MS: m/z 717.7 [M-Br] + .
实施例13Example 13
N-(3-(二甲基氨基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-19)N-(3-(Dimethylamino)propyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide (Compound A-19)
Figure PCTCN2020093248-appb-000049
Figure PCTCN2020093248-appb-000049
取δ-齐墩果酸(δ-OA，100mg，0.219mmol)溶于N,N-二甲基甲酰胺(8mL)中，加入三乙胺(122μL，0.876mmol)，室温搅拌下加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(108mg，0.285mmol)，反应1小时后加入3-二甲胺基丙胺(55μL，0.438mmol)，反应液升温至50℃。TLC检测反应完全后，用二氯甲烷(10mL)和水(5mL)处理反应液，水层用二氯甲烷(10mL×2)萃取，合并有机层，水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝30：1)纯化，得化合物V-1(白色固体，113mg，产率96％)。Dissolve δ-oleanolic acid (δ-OA, 100mg, 0.219mmol) in N,N-dimethylformamide (8mL), add triethylamine (122μL, 0.876mmol), add 2- with stirring at room temperature (7-oxybenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (108mg, 0.285mmol), react for 1 hour and add 3-dimethylaminopropylamine (55μL , 0.438mmol), the reaction solution was heated to 50°C. After the completion of the reaction detected by TLC, the reaction solution was treated with dichloromethane (10mL) and water (5mL), the aqueous layer was extracted with dichloromethane (10mL×2), the organic layers were combined, washed with water (5mL×2), and anhydrous sulfuric acid After drying with sodium, filtering, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane:methanol=30:1) to obtain compound V-1 (white solid, 113 mg, yield 96%).
将化合物V-1(113mg，0.209mmol)溶于无水吡啶(8mL)中，加入邻苯二甲酸酐(309mg，2.09mmol)和4-二甲氨基吡啶(26mg，0.209mmol)，反应液升温至115℃搅拌过夜。TLC检测反应完全后，加入二氯甲烷(20mL)稀释反应液，反应液依次用1N稀盐酸(10mL×3)和水(10mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物A-19(白色固体，121mg，产率84％)： 1H NMR(300MHz,CDCl 3)δ7.72(d,J＝7.2Hz,1H),7.55(d,J＝7.1Hz,1H),7.47–7.30(m,2H),6.36–6.27(m,1H),4.76–4.65(m,1H),3.46–3.18(m,2H),2.91–2.72(m,3H),2.62(s,6H),2.43(d,J＝14.2Hz,1H),2.39–2.30(m,1H),1.17(s,3H),0.98(s,3H),0.89(s,3H),0.87(s,6H),0.82(s,3H),0.72(s,3H).ESI-MS:m/z 689.6[M+H] +。 Compound V-1 (113 mg, 0.209 mmol) was dissolved in anhydrous pyridine (8 mL), phthalic anhydride (309 mg, 2.09 mmol) and 4-dimethylaminopyridine (26 mg, 0.209 mmol) were added, and the reaction solution was heated Stir at 115°C overnight. After the completion of the reaction detected by TLC, dichloromethane (20mL) was added to dilute the reaction solution. The reaction solution was washed with 1N dilute hydrochloric acid (10mL×3) and water (10mL×3) successively, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound A-19 (white solid, 121 mg, yield 84%): 1 H NMR (300MHz, CDCl 3 ) δ 7.72 (d, J =7.2Hz,1H),7.55(d,J=7.1Hz,1H),7.47–7.30(m,2H),6.36–6.27(m,1H),4.76–4.65(m,1H),3.46–3.18( m, 2H), 2.91–2.72 (m, 3H), 2.62 (s, 6H), 2.43 (d, J = 14.2 Hz, 1H), 2.39–2.30 (m, 1H), 1.17 (s, 3H), 0.98 (s, 3H), 0.89 (s, 3H), 0.87 (s, 6H), 0.82 (s, 3H), 0.72 (s, 3H). ESI-MS: m/z 689.6 [M+H] + .
实施例14Example 14
N-(3-(4-吗啉基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-20)N-(3-(4-morpholinyl)propyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide (compound A-20)
Figure PCTCN2020093248-appb-000050
Figure PCTCN2020093248-appb-000050
参照实施例13的方法，将3-二甲胺基丙胺替换成N-(3-氨丙基)吗啉，制得化合物A-20： 1H NMR(300MHz,CDCl 3)δ7.90–7.78(m,1H),7.72–7.59(m,1H),7.59–7.46(m,2H),6.40–6.25(m,1H),4.79–4.66(m,1H),4.13–3.93(m,4H),3.50–3.22(m,2H),3.21–3.04(m,2H),3.04–2.88(m,2H),2.81(d,J＝13.7Hz,1H),2.47(d,J＝13.8Hz,1H),2.35(d,J＝11.9Hz,1H),2.17–2.00(m,2H),1.19(s,3H),0.95(s,3H),0.89(s,6H),0.87(s,6H),0.73(s,3H). Referring to the method of Example 13, the 3-dimethylaminopropylamine was replaced with N-(3-aminopropyl)morpholine to prepare compound A-20: 1 H NMR (300MHz, CDCl 3 ) δ7.90-7.78 (m,1H), 7.72--7.59(m,1H), 7.59--7.46(m,2H), 6.40--6.25(m,1H), 4.79--4.66(m,1H), 4.13--3.93(m,4H) ,3.50–3.22(m,2H),3.21–3.04(m,2H),3.04–2.88(m,2H),2.81(d,J=13.7Hz,1H),2.47(d,J=13.8Hz,1H ), 2.35(d,J=11.9Hz,1H), 2.17–2.00(m,2H),1.19(s,3H),0.95(s,3H),0.89(s,6H),0.87(s,6H) ,0.73(s,3H).
实施例15Example 15
N-(3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰基)-吗啉(化合物A-27)N-(3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acyl)-morpholine (compound A-27)
Figure PCTCN2020093248-appb-000051
Figure PCTCN2020093248-appb-000051
参照实施例2的方法，将N,N-二甲基乙二胺替换成吗啉，制得化合物A-27： 1H NMR(300MHz,DMSO)δ7.94–7.85(m,1H),7.78–7.68(m,1H),7.63–7.49(m,2H),4.84–4.74(m,1H),3.80(s,4H),3.63(s,4H),2.77(d,J＝14.5Hz,1H),2.46(d,J＝13.2Hz,2H),1.19(s,3H),0.97(s,3H),0.92(s,3H),0.91(s,3H),0.90(s,3H),0.87(s,3H),0.79(s,3H).ESI-MS:m/z 672.5[M-H] -。 With reference to the method of Example 2, the N,N-dimethylethylenediamine was replaced with morpholine to obtain compound A-27: 1 H NMR (300MHz, DMSO) δ7.94-7.85(m,1H),7.78 –7.68(m,1H),7.63–7.49(m,2H), 4.84–4.74(m,1H), 3.80(s,4H),3.63(s,4H),2.77(d,J=14.5Hz,1H ), 2.46(d,J=13.2Hz,2H),1.19(s,3H),0.97(s,3H),0.92(s,3H),0.91(s,3H),0.90(s,3H),0.87 (s, 3H), 0.79 (s, 3H). ESI-MS: m/z 672.5 [MH] - .
实施例16Example 16
3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸2-(4-吗啉基)乙酯(化合物A-44)3β-(2-(2-Acetylaminoethoxyyl)benzoyl)oxy-oleanorane-13(18)-ene-28-acid 2-(4-morpholinyl)ethyl ester (Compound A -44)
Figure PCTCN2020093248-appb-000052
Figure PCTCN2020093248-appb-000052
参照实施例7的方法制得化合物III-5，取化合物III-5(120mg，0.174mmol)溶于N,N-二甲基甲酰胺(10mL)中，加入4-(2-溴乙基)吗啉氢溴酸盐(57mg，0.208mmol)和碳酸钾(70mg，0.522mmol)，室温搅拌反应。TLC检测反应完全后，用乙酸乙酯(20mL)和(10mL)处理反应液，水层用乙酸乙酯(10mL) 萃取，合并有机相，水(10mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物A-44(白色固体，87mg，产率62％)： 1H NMR(300MHz,CDCl 3)δ7.86–7.78(m,1H),7.69–7.63(m,1H),7.63–7.49(m,2H),6.64–6.47(m,1H),4.77–4.67(m,1H),4.44(t,J＝4.8Hz,2H),4.30–4.13(m,2H),3.77–3.55(m,6H),2.75(d,J＝14.0Hz,1H),2.59(t,J＝5.7Hz,2H),2.54–2.46(m,4H),2.42(d,J＝14.1Hz,1H),2.18(d,J＝6.9Hz,1H),2.03(s,3H),1.17(s,3H),0.96(s,6H),0.94(s,3H),0.93(s,3H),0.92(s,3H),0.74(s,3H).ESI-MS:m/z 825.6[M+Na] +。 The compound III-5 was prepared according to the method of Example 7. The compound III-5 (120mg, 0.174mmol) was dissolved in N,N-dimethylformamide (10mL), and 4-(2-bromoethyl) was added. Morpholine hydrobromide (57 mg, 0.208 mmol) and potassium carbonate (70 mg, 0.522 mmol) were reacted with stirring at room temperature. After the completion of the reaction detected by TLC, the reaction solution was treated with ethyl acetate (20 mL) and (10 mL), the aqueous layer was extracted with ethyl acetate (10 mL), the organic phases were combined, washed with water (10 mL×2), and dried with anhydrous sodium sulfate. Filter, concentrate the filtrate, and purify by silica gel column chromatography (dichloromethane:methanol=20:1) to obtain compound A-44 (white solid, 87mg, yield 62%): 1 H NMR (300MHz, CDCl 3 )δ7 .86–7.78(m,1H), 7.69–7.63(m,1H), 7.63–7.49(m,2H), 6.64–6.47(m,1H), 4.77–4.67(m,1H), 4.44(t, J=4.8Hz,2H), 4.30–4.13(m,2H), 3.77–3.55(m,6H), 2.75(d,J=14.0Hz,1H), 2.59(t,J=5.7Hz,2H), 2.54–2.46(m,4H),2.42(d,J=14.1Hz,1H), 2.18(d,J=6.9Hz,1H),2.03(s,3H),1.17(s,3H),0.96(s , 6H), 0.94 (s, 3H), 0.93 (s, 3H), 0.92 (s, 3H), 0.74 (s, 3H). ESI-MS: m/z 825.6 [M+Na] + .
实施例17Example 17
N-(2-(三甲基铵基)乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺溴化物(化合物A-66)N-(2-(Trimethylammonium)ethyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide bromide (Compound A-66)
Figure PCTCN2020093248-appb-000053
Figure PCTCN2020093248-appb-000053
参照实施例1的方法制得化合物A-66： 1H NMR(300MHz,DMSO-d 6)δ7.16–7.03(m,1H),4.31(d,J＝5.1Hz,1H),3.59–3.41(m,2H),3.08–2.92(m,2H),2.80(d,J＝12.4Hz,1H),2.45(d,J＝14.4Hz,1H),2.17(d,J＝13.4Hz,1H),1.15(s,3H),0.91(s,3H),0.90(s,6H),0.85(s,3H),0.73(s,3H),0.68(s,3H).ESI-MS:m/z 541.5[M-Br] +。 The compound A-66 was prepared by referring to the method of Example 1: 1 H NMR (300MHz, DMSO-d 6 ) δ 7.16–7.03 (m, 1H), 4.31 (d, J = 5.1 Hz, 1H), 3.59–3.41 (m,2H),3.08–2.92(m,2H), 2.80(d,J=12.4Hz,1H), 2.45(d,J=14.4Hz,1H), 2.17(d,J=13.4Hz,1H) ,1.15(s,3H),0.91(s,3H),0.90(s,6H),0.85(s,3H),0.73(s,3H),0.68(s,3H).ESI-MS: m/z 541.5[M-Br] + .
实施例18Example 18
N-(3-(三甲基铵基)丙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺溴化物(化合物A-67)N-(3-(Trimethylammonium)propyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide bromide (Compound A-67)
Figure PCTCN2020093248-appb-000054
Figure PCTCN2020093248-appb-000054
参照实施例1的方法，将化合物I-4替换成I-4’，制得化合物A-67： 1H NMR(300MHz,DMSO-d6)δ7.00–6.89(m,1H),4.31(d,J＝5.0Hz,1H),3.28–3.19(m,2H),3.19–3.09(m,2H),3.05(s,9H),2.79(d,J＝13.4Hz,1H),2.44(d,J＝13.9Hz,1H),2.18(d,J＝13.3Hz,1H),1.15(s,3H),0.90(s,9H),0.85(s,3H),0.72(s,3H),0.68(s,3H).ESI-MS:m/z 555.5[M-Br] +。 According to the method of Example 1, the compound I-4 was replaced with I-4' to obtain compound A-67: 1 H NMR (300MHz, DMSO-d6) δ7.00–6.89(m,1H), 4.31(d ,J=5.0Hz,1H), 3.28–3.19(m,2H), 3.19–3.09(m,2H),3.05(s,9H), 2.79(d,J=13.4Hz,1H), 2.44(d, J = 13.9Hz, 1H), 2.18 (d, J = 13.3Hz, 1H), 1.15 (s, 3H), 0.90 (s, 9H), 0.85 (s, 3H), 0.72 (s, 3H), 0.68 ( s, 3H). ESI-MS: m/z 555.5 [M-Br] + .
实施例19Example 19
N-(2-(二甲基氨基)乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-68)N-(2-(Dimethylamino)ethyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (Compound A-68)
Figure PCTCN2020093248-appb-000055
Figure PCTCN2020093248-appb-000055
参照实施例2的方法制得化合物A-68： 1H NMR(300MHz,CDCl 3)δ6.45(t,J＝4.7Hz,1H),3.45–3.31(m,2H),3.30–3.21(m,1H),2.93–2.80(m,1H),2.54–2.31(m,4H),2.23(s,6H),1.22(s,3H),1.03(s,3H),0.97(s,3H),0.94(s,3H),0.93(s,3H),0.81(s,3H),0.78(s,3H).ESI-MS:m/z 527.5[M+H] +。 The compound A-68 was prepared by referring to the method of Example 2: 1 H NMR (300MHz, CDCl 3 ) δ 6.45 (t, J = 4.7 Hz, 1H), 3.45-3.31 (m, 2H), 3.30-3.21 (m ,1H),2.93-2.80(m,1H),2.54-2.31(m,4H),2.23(s,6H),1.22(s,3H),1.03(s,3H),0.97(s,3H), 0.94 (s, 3H), 0.93 (s, 3H), 0.81 (s, 3H), 0.78 (s, 3H). ESI-MS: m/z 527.5 [M+H] + .
实施例20Example 20
N-(2-(1-哌啶基)乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-69)N-(2-(1-piperidinyl)ethyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (compound A-69)
Figure PCTCN2020093248-appb-000056
Figure PCTCN2020093248-appb-000056
参照实施例2的方法，将N,N-二甲基乙二胺替换成1-(2-氨基乙基)哌啶，制得化合物A-69： 1H NMR(300MHz,CDCl 3)δ6.71–6.26(m,1H),3.47–3.32(m,1H),3.32–3.16(m,2H),2.93–2.78(m,1H),2.55–2.22(m,8H),1.18(s,3H),0.99(s,3H),0.94(s,3H),0.90(s,3H),0.88(s,3H),0.77(s,3H),0.74(s,3H).ESI-MS:m/z 567.5[M+H] +。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with 1-(2-aminoethyl)piperidine to obtain compound A-69: 1 H NMR (300MHz, CDCl 3 )δ6. 71-6.26(m,1H), 3.47-3.32(m,1H), 3.32-3.16(m,2H), 2.93-2.78(m,1H), 2.55-2.22(m,8H), 1.18(s,3H) ),0.99(s,3H),0.94(s,3H),0.90(s,3H),0.88(s,3H),0.77(s,3H),0.74(s,3H).ESI-MS: m/ z 567.5[M+H] + .
实施例21Example 21
N-(2-(4-吗啉烷基)乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-70)N-(2-(4-morpholinoalkyl)ethyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (compound A-70)
Figure PCTCN2020093248-appb-000057
Figure PCTCN2020093248-appb-000057
参照实施例2的方法，将N,N-二甲基乙二胺替换成4-(2-氨基乙基)吗啉，制得化合物A-70： 1H NMR(300MHz,CDCl 3)δ6.50–6.40(m,1H),3.76–3.60(m,4H),3.45–3.29(m,2H),3.29–3.18(m,1H),2.90–2.79(m,1H),2.55–2.31(m,8H),1.19(s,3H),0.99(s,3H),0.94(s,3H),0.89(s,3H),0.87(s,3H),0.77(s,3H),0.74(s,3H).ESI-MS:m/z 569.5[M+H] +。 Referring to the method of Example 2, N,N-dimethylethylenediamine was replaced with 4-(2-aminoethyl)morpholine to obtain compound A-70: 1 H NMR (300MHz, CDCl 3 )δ6. 50-6.40 (m, 1H), 3.76-3.60 (m, 4H), 3.45-3.29 (m, 2H), 3.29-3.18 (m, 1H), 2.90-2.79 (m, 1H), 2.55-2.31 (m ,8H),1.19(s,3H),0.99(s,3H),0.94(s,3H),0.89(s,3H),0.87(s,3H),0.77(s,3H),0.74(s, 3H). ESI-MS: m/z 569.5 [M+H] + .
实施例22Example 22
N-(2-(4-甲基-1-哌嗪基)乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-73)N-(2-(4-Methyl-1-piperazinyl)ethyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (Compound A-73)
Figure PCTCN2020093248-appb-000058
Figure PCTCN2020093248-appb-000058
参照实施例2的方法，将N,N-二甲基乙二胺替换成1-(2-氨基乙基)-4-甲基哌嗪，制得化合物A-73： 1H NMR(300MHz,CDCl 3)δ6.53–6.44(m,1H),3.47–3.33(m,1H),3.34–3.18(m,2H),2.90–2.80(m,1H),2.64–2.33(m,12H),2.28(s,3H),1.19(s,3H),0.99(s,3H),0.94(s,3H),0.90(s,3H),0.89(s,3H),0.78(s,3H),0.75(s,3H).ESI-MS:m/z 582.5[M+H] +。 With reference to the method of Example 2, N,N-dimethylethylenediamine was replaced with 1-(2-aminoethyl)-4-methylpiperazine to obtain compound A-73: 1 H NMR (300MHz, CDCl 3 )δ6.53–6.44(m,1H), 3.47–3.33(m,1H), 3.34–3.18(m,2H), 2.90–2.80(m,1H), 2.64–2.33(m,12H), 2.28 (s, 3H), 1.19 (s, 3H), 0.99 (s, 3H), 0.94 (s, 3H), 0.90 (s, 3H), 0.89 (s, 3H), 0.78 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 582.5 [M+H] + .
实施例23Example 23
N-(3-(二甲基氨基)丙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-77)N-(3-(Dimethylamino)propyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (Compound A-77)
Figure PCTCN2020093248-appb-000059
Figure PCTCN2020093248-appb-000059
参照实施例2的方法，将N,N-二甲基乙二胺替换成3-二甲氨基丙胺，制得化合物A-77： 1H NMR(300MHz,CDCl 3)δ6.24(t,J＝5.1Hz,1H),3.46–3.32(m,1H),3.29–3.11(m,2H),2.87–2.73(m,1H),2.50–2.42(m,1H),2.41–2.35(m,1H),2.31(t,J＝7.1Hz,2H),2.22(s,6H),1.18(s,3H),0.99(s,3H),0.92(s,3H),0.91(s,3H),0.88(s,3H),0.77(s,3H),0.74(s,3H).ESI-MS:m/z 541.5[M+H] +。 With reference to the method of Example 2, N,N-dimethylethylenediamine was replaced with 3-dimethylaminopropylamine to obtain compound A-77: 1 H NMR (300MHz, CDCl 3 )δ6.24(t, J =5.1Hz,1H), 3.46–3.32(m,1H), 3.29–3.11(m,2H), 2.87–2.73(m,1H), 2.50–2.42(m,1H), 2.41–2.35(m,1H) ), 2.31(t, J=7.1Hz, 2H), 2.22(s, 6H), 1.18(s, 3H), 0.99(s, 3H), 0.92(s, 3H), 0.91(s, 3H), 0.88 (s, 3H), 0.77 (s, 3H), 0.74 (s, 3H). ESI-MS: m/z 541.5 [M+H] + .
实施例24Example 24
N-(3-(4-吗啉烷基)丙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-78)N-(3-(4-morpholinoalkyl)propyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (compound A-78)
Figure PCTCN2020093248-appb-000060
Figure PCTCN2020093248-appb-000060
参照实施例2的方法，将N,N-二甲基乙二胺替换成N-(3-氨丙基)吗啉，制得化合物A-78： 1H NMR(300MHz,CDCl 3)δ6.03(t,J＝5.7Hz,1H),3.71(d,J＝4.6Hz,4H),3.43–3.28(m,1H),3.28–3.14(m,2H),2.87–2.76(m,1H),2.53–2.30(m,8H),1.19(s,3H),0.99(s,3H),0.91(s,3H),0.90(s,3H),0.88(s,3H),0.78(s,3H),0.74(s,3H).ESI-MS:m/z 583.5[M+H] +。 With reference to the method of Example 2, N,N-dimethylethylenediamine was replaced with N-(3-aminopropyl)morpholine to obtain compound A-78: 1 H NMR (300MHz, CDCl 3 )δ6. 03(t,J=5.7Hz,1H), 3.71(d,J=4.6Hz,4H), 3.43–3.28(m,1H), 3.28–3.14(m,2H), 2.87–2.76(m,1H) ,2.53--2.30(m,8H),1.19(s,3H),0.99(s,3H),0.91(s,3H),0.90(s,3H),0.88(s,3H),0.78(s,3H) ), 0.74(s, 3H). ESI-MS: m/z 583.5[M+H] + .
实施例25Example 25
N-(2-(乙酰氨基)乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-83)N-(2-(Acetylamino)ethyl)-3β-Hydroxy-Olean-13(18)-ene-28-amide (Compound A-83)
Figure PCTCN2020093248-appb-000061
Figure PCTCN2020093248-appb-000061
参照实施例2的方法，将N,N-二甲基乙二胺替换成N-乙酰基乙二胺，制得化合物A-83： 1H NMR(300MHz,CDCl 3)δ6.49–6.35(m,1H),6.34–6.17(m,1H),3.55–3.28(m,4H),3.27–3.14(m,1H),2.88–2.74(m,1H),2.55–2.42(m,1H),2.42–2.30(m,1H),1.96(s,3H),1.19(s,3H),0.99(s,3H),0.91(s,6H),0.88(s,3H),0.77(s,3H),0.74(s,3H).ESI-MS:m/z 539.4[M-H] -。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with N-acetylethylenediamine to obtain compound A-83: 1 H NMR (300MHz, CDCl 3 )δ6.49–6.35( m,1H),6.34-6.17(m,1H),3.55--3.28(m,4H), 3.27-3.14(m,1H), 2.88-2.74(m,1H), 2.55-2.42(m,1H), 2.42-2.30(m,1H),1.96(s,3H),1.19(s,3H),0.99(s,3H),0.91(s,6H),0.88(s,3H),0.77(s,3H) ,0.74(s,3H). ESI-MS: m/z 539.4[MH] - .
实施例26Example 26
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-吗啉(化合物A-84)N-(3β-hydroxy-oleanorane-13(18)-ene-28-acyl)-morpholine (Compound A-84)
Figure PCTCN2020093248-appb-000062
Figure PCTCN2020093248-appb-000062
参照实施例2的方法，将N,N-二甲基乙二胺替换成吗啉，制得化合物A-84： 1H NMR(300MHz,CDCl 3)δ4.01–3.69(m,4H),3.68–3.52(m,4H),3.27–3.17(m,1H),2.81–2.70(m,1H),2.54–2.38(m,2H),1.17(s,3H),0.99(s,3H),0.92(s,3H),0.87(s,3H),0.86(s,3H),0.78(s,3H),0.77(s,3H).ESI-MS:m/z 548.4[M+Na] +。 With reference to the method of Example 2, N,N-dimethylethylenediamine was replaced with morpholine to obtain compound A-84: 1 H NMR (300MHz, CDCl 3 )δ4.01–3.69(m,4H), 3.68–3.52(m,4H), 3.27–3.17(m,1H), 2.81–2.70(m,1H), 2.54–2.38(m,2H), 1.17(s,3H), 0.99(s,3H), 0.92 (s, 3H), 0.87 (s, 3H), 0.86 (s, 3H), 0.78 (s, 3H), 0.77 (s, 3H). ESI-MS: m/z 548.4 [M+Na] + .
实施例27Example 27
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-硫代吗啉-1,1-二氧化物(化合物A-85)N-(3β-hydroxy-oleanorane-13(18)-ene-28-acyl)-thiomorpholine-1,1-dioxide (compound A-85)
Figure PCTCN2020093248-appb-000063
Figure PCTCN2020093248-appb-000063
参照实施例2的方法，将N,N-二甲基乙二胺替换成吗啉硫代吗啉-1,1-二氧化物，制得化合物A-85： 1H NMR(300MHz,CDCl 3)δ4.38–4.12(m,4H),3.28–3.17(m,1H),3.07–2.93(m,4H),2.81–2.70(m,1H),2.53–2.44(m,1H),2.44–2.33(m,1H),1.19(s,3H),0.99(s,3H),0.92(s,3H),0.87(s,3H),0.86(s,3H),0.78(s,3H),0.77(s,3H).ESI-MS:m/z 572.4[M-H] -。 With reference to the method of Example 2, N,N-dimethylethylenediamine was replaced with morpholine thiomorpholine-1,1-dioxide to obtain compound A-85: 1 H NMR (300MHz, CDCl 3 )δ 4.38--4.12(m,4H), 3.28--3.17(m,1H), 3.07--2.93(m,4H), 2.81--2.70(m,1H), 2.53--2.44(m,1H), 2.44-- 2.33(m,1H),1.19(s,3H),0.99(s,3H),0.92(s,3H),0.87(s,3H),0.86(s,3H),0.78(s,3H),0.77 (s, 3H). ESI-MS: m/z 572.4[MH] - .
实施例28Example 28
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-4-甲基哌嗪(化合物A-87)N-(3β-Hydroxy-oleanorane-13(18)-ene-28-acyl)-4-methylpiperazine (Compound A-87)
Figure PCTCN2020093248-appb-000064
Figure PCTCN2020093248-appb-000064
参照实施例2的方法，将N,N-二甲基乙二胺替换成N-甲基哌嗪，制得化合物A-87： 1H NMR(300MHz,CDCl 3)δ4.01–3.58(m,4H),3.28–3.19(m,1H),2.83–2.71(m,1H),2.54–2.40(m,2H),2.40–2.31(m,4H),2.29(s,3H),1.17(s,3H),0.99(s,3H),0.91(s,3H),0.89(s,3H),0.88(s,3H),0.78(s,3H),0.77(s,3H).ESI-MS:m/z 539.5[M+H] +。 Referring to the method of Example 2, N,N-dimethylethylenediamine was replaced with N-methylpiperazine to obtain compound A-87: 1 H NMR (300MHz, CDCl 3 ) δ4.01–3.58 (m ,4H), 3.28–3.19(m,1H), 2.83–2.71(m,1H), 2.54–2.40(m,2H), 2.40–2.31(m,4H), 2.29(s,3H), 1.17(s ,3H),0.99(s,3H),0.91(s,3H),0.89(s,3H),0.88(s,3H),0.78(s,3H),0.77(s,3H).ESI-MS: m/z 539.5 [M+H] + .
实施例29Example 29
N-(2-(二甲基氨基)乙基)-N-甲基-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-102)N-(2-(Dimethylamino)ethyl)-N-methyl-3β-hydroxy-oleanorane-13(18)-ene-28-amide (Compound A-102)
Figure PCTCN2020093248-appb-000065
Figure PCTCN2020093248-appb-000065
参照实施例2的方法，将N,N-二甲基乙二胺替换成N,N,N'-三甲基乙二胺，制得化合物A-102： 1H NMR(300MHz,CDCl 3)δ3.66–3.53(m,1H),3.53–3.38(m,1H),3.32–3.24(m,1H),3.22(s,3H),2.83–2.71(m,1H),2.57–2.36(m,4H),2.32(s,6H),1.19(s,3H),1.01(s,3H),0.96(s,3H),0.93(s,3H),0.89(s,3H),0.81(s,3H),0.79(s,3H).ESI-MS:m/z 541.5[M+H] +。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with N,N,N'-trimethylethylenediamine to obtain compound A-102: 1 H NMR (300MHz, CDCl 3 ) δ3.66--3.53(m,1H), 3.53--3.38(m,1H), 3.32--3.24(m,1H), 3.22(s,3H), 2.83--2.71(m,1H), 2.57--2.36(m ,4H),2.32(s,6H),1.19(s,3H),1.01(s,3H),0.96(s,3H),0.93(s,3H),0.89(s,3H),0.81(s, 3H), 0.79 (s, 3H). ESI-MS: m/z 541.5 [M+H] + .
实施例30Example 30
N-(2-(二乙基氨基)乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-103)N-(2-(Diethylamino)ethyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (Compound A-103)
Figure PCTCN2020093248-appb-000066
Figure PCTCN2020093248-appb-000066
参照实施例2的方法，将N,N-二甲基乙二胺替换成N,N-二乙基乙二胺，制得化合物A-103： 1H NMR(300MHz,CDCl 3)δ6.54–6.29(m,1H),3.50–3.32(m,1H),3.31–3.06(m,2H),2.87–2.78(m,1H),2.60–2.32(m,8H),1.18(s,3H),0.99(s,3H),0.93(s,3H),0.90(s,3H),0.88(s,3H),0.77(s,3H),0.74(s,3H).ESI-MS:m/z 555.4[M+H] +。 Refer to the method of Example 2 to replace N,N-dimethylethylenediamine with N,N-diethylethylenediamine to obtain compound A-103: 1 H NMR (300MHz, CDCl 3 )δ6.54 --6.29(m,1H), 3.50-3.32(m,1H), 3.31-3.06(m,2H), 2.87-2.78(m,1H), 2.60-2.32(m,8H), 1.18(s,3H) ,0.99(s,3H),0.93(s,3H),0.90(s,3H),0.88(s,3H),0.77(s,3H),0.74(s,3H).ESI-MS: m/z 555.4[M+H] + .
实施例31Example 31
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-哌啶(化合物A-104)N-(3β-Hydroxy-oleanorane-13(18)-ene-28-acyl)-piperidine (Compound A-104)
Figure PCTCN2020093248-appb-000067
Figure PCTCN2020093248-appb-000067
参照实施例2的方法，将N,N-二甲基乙二胺替换成哌啶，制得化合物A-104： 1H NMR(300MHz,CDCl 3)δ3.88–3.54(m,4H),3.31–3.22(m,1H),2.84–2.74(m,1H),2.54–2.37(m,2H),1.20(s,3H),1.02(s,3H),0.93(s,6H),0.91(s,3H),0.80(s,3H),0.80(s,3H).ESI-MS:m/z 546.4[M+Na] +。 Referring to the method of Example 2, replacing N,N-dimethylethylenediamine with piperidine, compound A-104 was prepared: 1 H NMR (300MHz, CDCl 3 ) δ 3.88-3.54 (m, 4H), 3.31–3.22(m,1H), 2.84–2.74(m,1H), 2.54–2.37(m,2H), 1.20(s,3H), 1.02(s,3H), 0.93(s,6H), 0.91( s, 3H), 0.80 (s, 3H), 0.80 (s, 3H). ESI-MS: m/z 546.4 [M+Na] + .
实施例32Example 32
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-哌嗪(化合物A-86)N-(3β-Hydroxy-oleanane-13(18)-ene-28-acyl)-piperazine (Compound A-86)
Figure PCTCN2020093248-appb-000068
Figure PCTCN2020093248-appb-000068
参照实施例2的方法，将N,N-二甲基乙二胺替换成哌嗪，制得化合物A-86： 1H NMR(300MHz,CDCl 3)δ3.87–3.64(m,3H),3.28–3.18(m,1H),2.88–2.78(m,3H),2.78–2.70(m,1H),2.45(d,J＝12.8Hz,2H),2.00–1.87(m,2H),1.17(s,3H),0.99(s,3H),0.91(s,3H),0.88(s,6H),0.77(s,6H).ESI-MS:m/z 525.6[M+H] +。 Referring to the method of Example 2, N,N-dimethylethylenediamine was replaced with piperazine to obtain compound A-86: 1 H NMR (300MHz, CDCl 3 ) δ 3.87–3.64 (m, 3H), 3.28–3.18(m,1H), 2.88–2.78(m,3H), 2.78–2.70(m,1H), 2.45(d, J=12.8Hz, 2H), 2.00–1.87(m,2H), 1.17( s, 3H), 0.99 (s, 3H), 0.91 (s, 3H), 0.88 (s, 6H), 0.77 (s, 6H). ESI-MS: m/z 525.6 [M+H] + .
实施例33Example 33
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-四氢吡咯(化合物A-88)N-(3β-Hydroxy-oleanorane-13(18)-ene-28-acyl)-tetrahydropyrrole (Compound A-88)
Figure PCTCN2020093248-appb-000069
Figure PCTCN2020093248-appb-000069
参照实施例2的方法，将N,N-二甲基乙二胺替换成四氢吡咯，制得化合物A-88： 1H NMR(300MHz,CDCl 3)δ3.66–3.49(m,4H),3.21(dd,J＝10.6,5.6Hz,1H),2.84–2.69(m,1H),2.48–2.38(m,1H),2.38–2.30(m,1H),1.16(s,3H),0.98(s,3H),0.94(s,3H),0.89(s,3H),0.87(s,3H),0.77(s,3H),0.76(s,3H).ESI-MS:m/z 532.5[M+Na] +。 Refer to the method of Example 2 to replace N,N-dimethylethylenediamine with tetrahydropyrrole to obtain compound A-88: 1 H NMR (300MHz, CDCl 3 ) δ 3.66–3.49 (m, 4H) ,3.21(dd,J=10.6,5.6Hz,1H), 2.84–2.69(m,1H), 2.48–2.38(m,1H), 2.38–2.30(m,1H), 1.16(s,3H), 0.98 (s,3H),0.94(s,3H),0.89(s,3H),0.87(s,3H),0.77(s,3H),0.76(s,3H).ESI-MS:m/z 532.5[ M+Na] + .
实施例34Example 34
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-(3S)-羟基-四氢吡咯(化合物A-91)N-(3β-hydroxy-oleanorane-13(18)-ene-28-acyl)-(3S)-hydroxy-tetrahydropyrrole (Compound A-91)
Figure PCTCN2020093248-appb-000070
Figure PCTCN2020093248-appb-000070
参照实施例2的方法，将N,N-二甲基乙二胺替换成(S)-3-吡咯烷醇，制得化合物A-91： 1H NMR(300MHz,CDCl 3)δ5.29(s,1H),4.49–4.36(m,1H),3.91–3.72(m,1H),3.73–3.54(m,3H),3.22(dd,J＝9.6,4.9Hz,1H),2.76(d,J＝14.9Hz,1H),2.50–2.39(m,1H),2.39–2.29(m,1H),2.22–2.09(m,1H),1.16(s,3H),0.98(s,3H),0.94(s,3H),0.90(s,3H),0.87(s,3H),0.77(s,3H),0.76(s,3H).ESI-MS:m/z 526.4[M+H] +。 Referring to the method of Example 2, N,N-dimethylethylenediamine was replaced with (S)-3-pyrrolidinol to obtain compound A-91: 1 H NMR (300MHz, CDCl 3 )δ5.29( s,1H), 4.49–4.36(m,1H), 3.91–3.72(m,1H), 3.73–3.54(m,3H), 3.22(dd,J=9.6,4.9Hz,1H), 2.76(d, J=14.9Hz,1H), 2.50–2.39(m,1H), 2.39–2.29(m,1H), 2.22–2.09(m,1H), 1.16(s,3H), 0.98(s,3H), 0.94 (s, 3H), 0.90 (s, 3H), 0.87 (s, 3H), 0.77 (s, 3H), 0.76 (s, 3H). ESI-MS: m/z 526.4 [M+H] + .
实施例35Example 35
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-(3R)-羟基-四氢吡咯(化合物A-92)N-(3β-hydroxy-oleanorane-13(18)-ene-28-acyl)-(3R)-hydroxy-tetrahydropyrrole (Compound A-92)
Figure PCTCN2020093248-appb-000071
Figure PCTCN2020093248-appb-000071
参照实施例2的方法，将N,N-二甲基乙二胺替换成(R)-3-吡咯烷醇，制得化合物A-92： 1H NMR(300MHz,CDCl 3)δ4.50–4.34(m,1H),3.80–3.66(m,2H),3.68–3.60(m,2H),3.30–3.12(m,1H),2.75(d,J＝14.8Hz,1H),2.48–2.37(m,1H),2.38–2.26(m,2H),1.15(s,3H),0.97(s,3H),0.95(s,3H),0.89(s,3H),0.86(s,3H),0.77(s,3H),0.75(s,3H).ESI-MS:m/z 526.4[M+H] +。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with (R)-3-pyrrolidinol to obtain compound A-92: 1 H NMR (300MHz, CDCl 3 ) δ4.50– 4.34(m,1H), 3.80–3.66(m,2H), 3.68–3.60(m,2H), 3.30–3.12(m,1H), 2.75(d,J=14.8Hz,1H), 2.48–2.37( m,1H),2.38--2.26(m,2H),1.15(s,3H),0.97(s,3H),0.95(s,3H),0.89(s,3H),0.86(s,3H),0.77 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 526.4 [M+H] + .
实施例36Example 36
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-(2R)-羧基-四氢吡咯(化合物A-93)N-(3β-hydroxy-oleanorane-13(18)-ene-28-acyl)-(2R)-carboxy-tetrahydropyrrole (compound A-93)
Figure PCTCN2020093248-appb-000072
Figure PCTCN2020093248-appb-000072
参照实施例2的方法，将N,N-二甲基乙二胺替换成D-脯氨酸苄酯盐酸盐，制得化合物VI-1。According to the method of Example 2, N,N-dimethylethylenediamine was replaced with D-proline benzyl ester hydrochloride to obtain compound VI-1.
取化合物VI-1(120mg，0.186mmol)溶于四氢呋喃(1mL)和甲醇(1mL)的混合溶液中，加入2N氢氧化钠溶液(1mL)，80℃下反应2小时。TLC检测反应完全后，减压蒸除溶剂，用1N盐酸溶液调节pH至弱酸性，二氯甲烷萃取(5mL×2)，水洗(5mL×2)有机相，饱和食盐水洗涤(5mL×2)，无水硫酸钠干燥，减压蒸除溶剂，残余物用二氯甲烷和乙腈的混合溶剂(5.5mL，v：v＝10：1)于超声中处理，析出白色固体，抽滤，滤饼用二氯甲烷和乙腈的混合溶剂(5.5mL，v：v＝10：1)洗涤两次，红外下干燥，得化合物A-93(白色固体， 88mg，产率85％)： 1H NMR(300MHz,CDCl 3)δ4.70(q,J＝3.8Hz,1H),3.94–3.79(m,1H),3.72–3.59(m,1H),3.25(dd,J＝10.8,5.4Hz,1H),2.79(dd,J＝15.5,4.0Hz,1H),2.53–2.40(m,1H),2.38–2.21(m,2H),1.19(s,3H),1.01(s,3H),0.94(s,6H),0.89(s,3H),0.81(s,3H),0.78(s,3H).ESI-MS:m/z 552.4[M-H] -。 The compound VI-1 (120 mg, 0.186 mmol) was dissolved in a mixed solution of tetrahydrofuran (1 mL) and methanol (1 mL), 2N sodium hydroxide solution (1 mL) was added, and the reaction was carried out at 80° C. for 2 hours. After the completion of the reaction detected by TLC, the solvent was evaporated under reduced pressure, the pH was adjusted to weak acidity with 1N hydrochloric acid solution, extracted with dichloromethane (5mL×2), the organic phase was washed with water (5mL×2), and washed with saturated brine (5mL×2) , Dried over anhydrous sodium sulfate, distilled off the solvent under reduced pressure, the residue was treated with a mixed solvent of dichloromethane and acetonitrile (5.5mL, v:v=10:1) under ultrasound, a white solid precipitated, filtered with suction, and filter cake Wash twice with a mixed solvent of dichloromethane and acetonitrile (5.5mL, v:v=10:1), and dry under infrared to obtain compound A-93 (white solid, 88mg, yield 85%): 1 H NMR( 300MHz, CDCl 3 )δ4.70(q,J=3.8Hz,1H),3.94-3.79(m,1H),3.72-3.59(m,1H), 3.25(dd,J=10.8,5.4Hz,1H) ,2.79(dd,J=15.5,4.0Hz,1H),2.53–2.40(m,1H),2.38–2.21(m,2H),1.19(s,3H),1.01(s,3H),0.94(s , 6H), 0.89 (s, 3H), 0.81 (s, 3H), 0.78 (s, 3H). ESI-MS: m/z 552.4[MH] - .
实施例37Example 37
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-(2S)-羧基-四氢吡咯(化合物A-94)N-(3β-hydroxy-oleanorane-13(18)-ene-28-acyl)-(2S)-carboxy-tetrahydropyrrole (Compound A-94)
Figure PCTCN2020093248-appb-000073
Figure PCTCN2020093248-appb-000073
参照实施例36的方法，将D-脯氨酸苄酯盐酸盐替换成L-脯氨酸苄酯盐酸盐，制得化合物A-94： 1H NMR(300MHz,CDCl 3)δ4.77–4.62(m,1H),3.90–3.75(m,1H),3.74–3.56(m,1H),3.34–3.18(m,1H),2.78(d,J＝17.1Hz,1H),2.53–2.43(m,1H),2.42–2.33(m,1H),2.28–2.16(m,1H),1.19(s,3H),1.00(s,3H),0.94(s,3H),0.92(s,3H),0.89(s,3H),0.80(s,3H),0.78(s,3H).ESI-MS:m/z 576.53[M+Na] +。 Referring to the method of Example 36, D-proline benzyl ester hydrochloride was replaced with L-proline benzyl ester hydrochloride to obtain compound A-94: 1 H NMR (300MHz, CDCl 3 ) δ4.77 –4.62(m,1H),3.90–3.75(m,1H), 3.74–3.56(m,1H), 3.34–3.18(m,1H), 2.78(d,J=17.1Hz,1H), 2.53–2.43 (m,1H),2.42-2.33(m,1H),2.28-2.16(m,1H),1.19(s,3H),1.00(s,3H),0.94(s,3H),0.92(s,3H) ), 0.89 (s, 3H), 0.80 (s, 3H), 0.78 (s, 3H). ESI-MS: m/z 576.53 [M+Na] + .
实施例38Example 38
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-吖丁啶(化合物A-95)N-(3β-Hydroxy-oleanane-13(18)-ene-28-acyl)-azetidine (Compound A-95)
Figure PCTCN2020093248-appb-000074
Figure PCTCN2020093248-appb-000074
参照实施例2的方法，将N,N-二甲基乙二胺替换成杂氮环丁烷，制得化合物A-95： 1H NMR(300MHz,CDCl 3)δ4.48–3.89(m,4H),3.30–3.14(m,1H),2.81–2.66(m,1H),2.48–2.34(m,1H),2.26–2.07(m,3H),1.14(s,3H),0.97(s,3H),0.95(s,3H),0.91(s,3H),0.86(s,3H),0.75(s,3H),0.74(s,3H).ESI-MS:m/z 518.5[M+Na] +。 According to the method of Example 2, N,N-dimethylethylenediamine was replaced with azacyclobutane to obtain compound A-95: 1 H NMR (300MHz, CDCl 3 ) δ 4.48–3.89 (m, 4H), 3.30--3.14(m,1H), 2.81--2.66(m,1H), 2.48--2.34(m,1H), 2.26--2.07(m,3H), 1.14(s,3H), 0.97(s, 3H),0.95(s,3H),0.91(s,3H),0.86(s,3H),0.75(s,3H),0.74(s,3H).ESI-MS:m/z 518.5(M+Na ] + .
实施例39Example 39
N-(3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰基)-硫代吗啉-1,1-二氧化物(化合物A-28)N-(3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acyl)-thiomorpholine-1,1-dioxide (compound A-28 )
Figure PCTCN2020093248-appb-000075
Figure PCTCN2020093248-appb-000075
参照实施例2的方法，将N,N-二甲基乙二胺替换成硫代吗啉-1,1-二氧化物，制得化合物A-28： 1H NMR(300MHz,CDCl 3)δ7.96–7.86(m,1H),7.81–7.70(m,1H),7.65–7.52(m,2H),4.85–4.73(m,1H),4.37–4.12(m,4H),3.08–2.92(m, 4H),2.77(d,J＝13.3Hz,1H),2.49(d,J＝13.2Hz,1H),2.44–2.32(m,1H),1.20(s,3H),0.97(s,3H),0.90(s,9H),0.86(s,3H),0.79(s,3H).ESI-MS:m/z 720.5[M-H] -。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with thiomorpholine-1,1-dioxide to obtain compound A-28: 1 H NMR (300MHz, CDCl 3 )δ7 .96–7.86(m,1H), 7.81–7.70(m,1H), 7.65–7.52(m,2H), 4.85–4.73(m,1H), 4.37–4.12(m,4H), 3.08–2.92( m, 4H), 2.77 (d, J = 13.3 Hz, 1H), 2.49 (d, J = 13.2 Hz, 1H), 2.44–2.32 (m, 1H), 1.20 (s, 3H), 0.97 (s, 3H) ), 0.90 (s, 9H), 0.86 (s, 3H), 0.79 (s, 3H). ESI-MS: m/z 720.5 [MH] - .
实施例40Example 40
N-(2-(1,1-二氧代硫代吗啉基)乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-74)N-(2-(1,1-dioxothiomorpholinyl)ethyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (compound A-74)
Figure PCTCN2020093248-appb-000076
Figure PCTCN2020093248-appb-000076
参照实施例13的方法，将3-二甲氨基丙胺替换成4-(2-氨乙基)硫代吗啉-1,1-二氧化物，制得化合物A-74： 1H NMR(300MHz,CDCl 3)δ6.27–6.11(m,1H),3.48–3.29(m,2H),3.29–3.16(m,1H),3.13–2.91(m,8H),2.82(d,J＝13.0Hz,1H),2.60(t,J＝5.7Hz,2H),2.47(d,J＝13.5Hz,1H),2.43–2.31(m,1H),1.20(s,3H),0.99(s,3H),0.92(s,3H),0.90(s,3H),0.87(s,3H),0.76(s,3H),0.74(s,3H).ESI-MS:m/z 615.5[M-H] -。 Referring to the method in Example 13, the 3-dimethylaminopropylamine was replaced with 4-(2-aminoethyl)thiomorpholine-1,1-dioxide to obtain compound A-74: 1 H NMR (300MHz ,CDCl 3 )δ6.27–6.11(m,1H), 3.48–3.29(m,2H), 3.29–3.16(m,1H), 3.13–2.91(m,8H), 2.82(d,J=13.0Hz ,1H), 2.60(t,J=5.7Hz,2H),2.47(d,J=13.5Hz,1H),2.43-2.31(m,1H),1.20(s,3H),0.99(s,3H) , 0.92 (s, 3H), 0.90 (s, 3H), 0.87 (s, 3H), 0.76 (s, 3H), 0.74 (s, 3H). ESI-MS: m/z 615.5 [MH] - .
实施例41Example 41
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-81)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (compound A-81)
Figure PCTCN2020093248-appb-000077
Figure PCTCN2020093248-appb-000077
参照实施例13的方法，将3-二甲氨基丙胺替换成4-(3-氨丙基)硫代吗啉-1,1-二氧化物，制得化合物A-81： 1H NMR(300MHz,CDCl 3)δ6.05–5.95(m,1H),3.38–3.17(m,3H),3.12–2.90(m,8H),2.81(d,J＝12.5Hz,1H),2.59–2.32(m,4H),1.19(s,3H),0.99(s,3H),0.90(s,6H),0.88(s,3H),0.77(s,3H),0.73(s,3H).ESI-MS:m/z 629.5[M-H] -。 Referring to the method in Example 13, the 3-dimethylaminopropylamine was replaced with 4-(3-aminopropyl)thiomorpholine-1,1-dioxide to obtain compound A-81: 1 H NMR (300MHz ,CDCl 3 )δ6.05–5.95(m,1H), 3.38–3.17(m,3H), 3.12–2.90(m,8H), 2.81(d,J=12.5Hz,1H), 2.59–2.32(m ,4H),1.19(s,3H),0.99(s,3H),0.90(s,6H),0.88(s,3H),0.77(s,3H),0.73(s,3H).ESI-MS: m/z 629.5[MH] - .
实施例42Example 42
N-(2-(1,1-二氧代硫代吗啉基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-9)N-(2-(1,1-dioxothiomorpholinyl)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28- Amide (Compound A-9)
Figure PCTCN2020093248-appb-000078
Figure PCTCN2020093248-appb-000078
参照实施例13的方法，将3-二甲氨基丙胺替换成4-(2-氨乙基)硫代吗啉-1,1-二氧化物，制得化合物A-9： 1H NMR(300MHz,DMSO-d 6)δ13.20(s,1H),7.79–7.69(m,1H),7.69–7.59(m,3H),6.63–6.52(m,1H),4.73–4.60(m,1H),3.29– 3.15(m,2H),3.12–2.98(m,4H),2.98–2.87(m,4H),2.83(d,J＝15.0Hz,1H),2.62–2.53(m,2H),2.47(d,J＝14.2Hz,1H),1.24–1.17(m,3H),0.94(s,6H),0.92(s,6H),0.85(s,3H),0.74(s,3H).ESI-MS:m/z 763.6[M-H] -。 Referring to the method in Example 13, the 3-dimethylaminopropylamine was replaced with 4-(2-aminoethyl)thiomorpholine-1,1-dioxide to obtain compound A-9: 1 H NMR (300MHz ,DMSO-d 6 )δ13.20(s,1H), 7.79-7.69(m,1H), 7.69-7.59(m,3H), 6.63-6.52(m,1H), 4.73-4.60(m,1H) , 3.29– 3.15(m, 2H), 3.12–2.98(m, 4H), 2.98–2.87(m, 4H), 2.83(d, J = 15.0Hz, 1H), 2.62–2.53(m, 2H), 2.47 (d,J=14.2Hz,1H),1.24–1.17(m,3H),0.94(s,6H),0.92(s,6H),0.85(s,3H),0.74(s,3H).ESI- MS: m/z 763.6 [MH] - .
实施例43Example 43
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-23)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28- Amide (Compound A-23)
Figure PCTCN2020093248-appb-000079
Figure PCTCN2020093248-appb-000079
参照实施例13的方法，将3-二甲氨基丙胺替换成4-(3-氨丙基)硫代吗啉-1,1-二氧化物，制得化合物A-23： 1H NMR(300MHz,methanol-d 4)δ7.82–7.74(m,1H),7.74–7.66(m,1H),7.66–7.57(m,2H),6.86–6.77(m,1H),4.80–4.67(m,1H),3.42–3.23(m,2H),3.18–3.07(m,4H),3.07–2.97(m,4H),2.97–2.86(m,1H),2.64–2.49(m,3H),2.31(d,J＝12.8Hz,1H),1.28(s,3H),1.03(s,6H),1.00(s,3H),0.95(s,3H),0.94(s,3H),0.81(s,3H).ESI-MS:m/z 777.6[M-H] -。 Referring to the method of Example 13, the 3-dimethylaminopropylamine was replaced with 4-(3-aminopropyl)thiomorpholine-1,1-dioxide to obtain compound A-23: 1 H NMR (300MHz ,methanol-d 4 )δ7.82–7.74(m,1H), 7.74–7.66(m,1H), 7.66–7.57(m,2H), 6.86–6.77(m,1H), 4.80–4.67(m, 1H), 3.42–3.23(m, 2H), 3.18–3.07(m, 4H), 3.07–2.97(m, 4H), 2.97–2.86(m, 1H), 2.64–2.49(m, 3H), 2.31( d,J=12.8Hz,1H),1.28(s,3H),1.03(s,6H),1.00(s,3H),0.95(s,3H),0.94(s,3H),0.81(s,3H) ). ESI-MS: m/z 777.6[MH] - .
实施例44Example 44
N-(3-(1-四氢吡咯基)丙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-80)N-(3-(1-Tetrahydropyrrolyl)propyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (Compound A-80)
Figure PCTCN2020093248-appb-000080
Figure PCTCN2020093248-appb-000080
参照实施例13的方法，将3-二甲氨基丙胺替换成1-(3-氨基丙基)吡咯烷，制得化合物A-80： 1H NMR(300MHz,CDCl 3)δ6.22–6.12(m,1H),3.50–3.34(m,1H),3.29–3.15(m,2H),2.87–2.75(m,1H),2.61–2.37(m,8H),1.21(s,3H),1.18(s,3H),0.99(s,3H),0.91(s,3H),0.88(s,3H),0.77(s,3H),0.73(s,3H).ESI-MS:m/z 567.5[M+H] +。 According to the method of Example 13, the 3-dimethylaminopropylamine was replaced with 1-(3-aminopropyl)pyrrolidine to obtain compound A-80: 1 H NMR (300MHz, CDCl 3 )δ6.22-6.12( m,1H), 3.50--3.34(m,1H), 3.29--3.15(m,2H), 2.87--2.75(m,1H), 2.61--2.37(m,8H), 1.21(s,3H), 1.18( s,3H),0.99(s,3H),0.91(s,3H),0.88(s,3H),0.77(s,3H),0.73(s,3H).ESI-MS:m/z 567.5(M +H] + .
实施例45Example 45
N-(3-(4-甲基-1-哌嗪基)丙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-79)N-(3-(4-Methyl-1-piperazinyl)propyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (Compound A-79)
Figure PCTCN2020093248-appb-000081
Figure PCTCN2020093248-appb-000081
参照实施例13的方法，将3-二甲氨基丙胺替换成1-(3-氨丙基)-4-甲基哌嗪，制得化合物A-79： 1H NMR(300MHz,CDCl 3)δ6.09–5.96(m,1H),3.41–3.28(m,1H),3.27–3.12(m,2H),2.86–2.72(m,1H),2.65–2.31(m,10H),2.28(s,3H),2.02–1.79(m,2H),1.18(s,6H),0.98(s,3H),0.88(s,6H),0.76(s,3H),0.72(s,3H).ESI-MS:m/z 596.5[M+H] +。 Referring to the method of Example 13, the 3-dimethylaminopropylamine was replaced with 1-(3-aminopropyl)-4-methylpiperazine to obtain compound A-79: 1 H NMR (300MHz, CDCl 3 )δ6 .09–5.96(m,1H),3.41–3.28(m,1H), 3.27–3.12(m,2H), 2.86–2.72(m,1H), 2.65–2.31(m,10H), 2.28(s, 3H),2.02--1.79(m,2H),1.18(s,6H),0.98(s,3H),0.88(s,6H),0.76(s,3H),0.72(s,3H).ESI-MS :m/z 596.5[M+H] + .
实施例46Example 46
N-(3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺)四氢吡咯(化合物A-31)N-(3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide)tetrahydropyrrole (Compound A-31)
Figure PCTCN2020093248-appb-000082
Figure PCTCN2020093248-appb-000082
参照实施例2的方法，将N,N-二甲基乙二胺替换成四氢吡咯，制得化合物A-31： 1H NMR(300MHz,methanol-d 4)δ7.81–7.65(m,2H),7.65–7.51(m,2H),4.80–4.70(m,1H),3.81–3.44(m,4H),2.86(d,J＝15.1Hz,1H),2.52(d,J＝13.4Hz,1H),2.37(d,J＝13.1Hz,1H),1.27(s,3H),1.03(s,3H),1.01(s,6H),0.95(s,6H),0.85(s,3H).ESI-MS:m/z 656.5[M-H] -。 According to the method of Example 2, N,N-dimethylethylenediamine was replaced with tetrahydropyrrole to obtain compound A-31: 1 H NMR (300MHz, methanol-d 4 )δ7.81–7.65(m, 2H), 7.65–7.51(m,2H), 4.80–4.70(m,1H), 3.81–3.44(m,4H), 2.86(d,J=15.1Hz,1H), 2.52(d,J=13.4Hz ,1H),2.37(d,J=13.1Hz,1H),1.27(s,3H),1.03(s,3H),1.01(s,6H),0.95(s,6H),0.85(s,3H) .ESI-MS: m/z 656.5[MH] - .
实施例47Example 47
N-(2-(乙酰氨基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-64)N-(2-(Acetylamino)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide (Compound A-64)
Figure PCTCN2020093248-appb-000083
Figure PCTCN2020093248-appb-000083
参照实施例13的方法，将3-二甲氨基丙胺替换成N-乙酰基乙二胺，制得化合物A-64： 1H NMR(300MHz,CDCl 3)δ7.93–7.77(m,1H),7.77–7.62(m,1H),7.60–7.46(m,2H),6.62–6.45(m,1H),6.38–6.21(m,1H),4.82–4.66(m,1H),3.57–3.20(m,4H),2.82(d,J＝15.1Hz,1H),2.46(d,J＝13.3Hz,1H),2.36(d,J＝11.6Hz,1H),1.97(s,3H),1.19(s,3H),0.95(s,3H),0.90(s,12H),0.74(s,3H).ESI-MS:m/z 687.5[M-H] -。 Referring to the method of Example 13, the 3-dimethylaminopropylamine was replaced with N-acetylethylenediamine to obtain compound A-64: 1 H NMR (300MHz, CDCl 3 ) δ7.93-7.77 (m, 1H) ,7.77–7.62(m,1H), 7.60–7.46(m,2H), 6.62–6.45(m,1H), 6.38–6.21(m,1H), 4.82–4.66(m,1H), 3.57–3.20( m, 4H), 2.82 (d, J = 15.1 Hz, 1H), 2.46 (d, J = 13.3 Hz, 1H), 2.36 (d, J = 11.6 Hz, 1H), 1.97 (s, 3H), 1.19 ( s, 3H), 0.95 (s, 3H), 0.90 (s, 12H), 0.74 (s, 3H). ESI-MS: m/z 687.5[MH] - .
实施例48Example 48
N-(2-(乙酰氨基)乙基)-3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-65)N-(2-(Acetylamino)ethyl)-3β-(2-(2-Acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-13(18)-ene-28-amide( Compound A-65)
Figure PCTCN2020093248-appb-000084
Figure PCTCN2020093248-appb-000084
参照实施例7的方法，将N-(2-氨基乙基)吗啉替换成N-乙酰基乙二胺，制得化合物A-65： 1H NMR(300MHz,CDCl 3)δ7.85–7.77(m,1H),7.70–7.63(m,1H),7.62–7.50(m,2H),6.60–6.47(m,1H),6.43–6.34(m,1H),6.33–6.23(m,1H),4.76–4.67(m,1H),4.45(t,J＝4.7Hz,2H),3.71–3.58(m,2H),3.49–3.26(m,4H),2.91–2.76(m,1H),2.48(d,J＝14.3Hz,1H),2.42–2.32(m,1H),2.03(s,3H),1.96(s,3H),1.21(s,3H),0.97(s,3H),0.96(s,6H),0.93(s,3H),0.91(s,3H),0.75(s,3H).ESI-MS:m/z 796.6[M+Na] +。 Referring to the method of Example 7, N-(2-aminoethyl)morpholine was replaced with N-acetylethylenediamine to obtain compound A-65: 1 H NMR (300MHz, CDCl 3 ) δ7.85-7.77 (m,1H), 7.70--7.63(m,1H), 7.62--7.50(m,2H), 6.60--6.47(m,1H), 6.43--6.34(m,1H), 6.33--6.23(m,1H) ,4.76–4.67(m,1H), 4.45(t,J=4.7Hz,2H), 3.71–3.58(m,2H), 3.49–3.26(m,4H), 2.91–2.76(m,1H), 2.48 (d,J=14.3Hz,1H),2.42–2.32(m,1H),2.03(s,3H),1.96(s,3H),1.21(s,3H),0.97(s,3H),0.96( s, 6H), 0.93 (s, 3H), 0.91 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 796.6 [M+Na] + .
实施例49Example 49
N-(2-(1-四氢吡咯基)乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-71)N-(2-(1-Tetrahydropyrrolyl)ethyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (Compound A-71)
Figure PCTCN2020093248-appb-000085
Figure PCTCN2020093248-appb-000085
参照实施例13的方法，将3-二甲氨基丙胺替换成1-(2-氨乙基)吡咯烷，制得化合物A-71： 1H NMR(300MHz,CDCl 3)δ6.59–6.45(m,1H),3.46–3.17(m,3H),2.89–2.74(m,1H),2.60–2.30(m,7H),1.18(s,6H),0.99(s,3H),0.93(s,3H),0.89(s,3H),0.77(s,3H),0.74(s,3H).ESI-MS:m/z 553.5[M+H] +。 According to the method of Example 13, the 3-dimethylaminopropylamine was replaced with 1-(2-aminoethyl)pyrrolidine to obtain compound A-71: 1 H NMR (300MHz, CDCl 3 )δ6.59–6.45( m,1H), 3.46–3.17(m,3H), 2.89–2.74(m,1H), 2.60–2.30(m,7H), 1.18(s,6H), 0.99(s,3H), 0.93(s, 3H), 0.89 (s, 3H), 0.77 (s, 3H), 0.74 (s, 3H). ESI-MS: m/z 553.5 [M+H] + .
实施例50Example 50
1-甲基-4-(3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺)哌嗪(化合物A-30)1-Methyl-4-(3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide)piperazine (Compound A-30)
Figure PCTCN2020093248-appb-000086
Figure PCTCN2020093248-appb-000086
参照实施例2的方法，将N,N-二甲基乙二胺替换成N-甲基哌嗪，制得化合物A-30： 1H NMR(300MHz,methanol-d 4)δ7.79–7.72(m,1H),7.72–7.65(m,1H),7.65–7.54(m,2H),4.74(dd,J＝10.7,5.1Hz,1H),4.28–3.84(m,4H),3.28–3.12(m,4H),2.89(s,3H),2.84(d,J＝12.6Hz,1H),2.57(d,J＝13.3Hz,1H),2.38(d,J＝12.6Hz,1H),1.27(s,3H),1.00(s,3H),0.99(s,3H),0.97(s,3H),0.95(s,3H),0.93(s,3H),0.85(s,3H).ESI-MS:m/z 687.5[M+H] +。 Refer to the method of Example 2 to replace N,N-dimethylethylenediamine with N-methylpiperazine to obtain compound A-30: 1 H NMR (300MHz, methanol-d 4 )δ7.79–7.72 (m,1H),7.72–7.65(m,1H), 7.65–7.54(m,2H), 4.74(dd,J=10.7,5.1Hz,1H), 4.28–3.84(m,4H), 3.28–3.12 (m, 4H), 2.89 (s, 3H), 2.84 (d, J = 12.6 Hz, 1H), 2.57 (d, J = 13.3 Hz, 1H), 2.38 (d, J = 12.6 Hz, 1H), 1.27 (s,3H),1.00(s,3H),0.99(s,3H),0.97(s,3H),0.95(s,3H),0.93(s,3H),0.85(s,3H).ESI- MS: m/z 687.5 [M+H] + .
实施例51Example 51
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-(3S)-氨基-四氢吡咯(化合物A-89)N-(3β-hydroxy-oleanorane-13(18)-ene-28-acyl)-(3S)-amino-tetrahydropyrrole (Compound A-89)
Figure PCTCN2020093248-appb-000087
Figure PCTCN2020093248-appb-000087
参照实施例2的方法，将N,N-二甲基乙二胺替换成(S)-3-N-苄氧羰基氨基吡咯烷，制得化合物VII-1。According to the method of Example 2, N,N-dimethylethylenediamine was replaced with (S)-3-N-benzyloxycarbonylaminopyrrolidine to obtain compound VII-1.
取化合物VII-1(200mg，0.303mmol)溶于四氢呋喃(10mL)中，加入10％钯碳(20mg)，氢气氛围下室温搅拌过夜。TLC检测反应完全后，过滤硅藻土，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝10：1)纯化，得化合物A-89(白色固体，135mg，产率85％)： 1H NMR(300MHz,CDCl 3)δ3.84–3.75(m,1H),3.74–3.66(m,1H),3.66–3.57(m,1H),3.51(p,J＝6.0Hz,1H),3.24(td,J＝10.6,5.9Hz,2H),2.82–2.69(m,1H),2.48–2.38(m,1H),2.39–2.29(m,1H),2.04(dt,J＝12.4,6.6Hz,1H),1.16(s,3H),0.98(s,3H),0.95(s,3H),0.90(s,3H),0.87(s,3H),0.77(s,3H),0.76(s,3H).ESI-MS:m/z 525.5[M+H] +。 The compound VII-1 (200 mg, 0.303 mmol) was dissolved in tetrahydrofuran (10 mL), 10% palladium on carbon (20 mg) was added, and the mixture was stirred at room temperature overnight under a hydrogen atmosphere. After the completion of the reaction detected by TLC, diatomaceous earth was filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 10:1) to obtain compound A-89 (white solid, 135 mg, yield 85%): 1 H NMR (300MHz, CDCl 3 ) δ 3.84–3.75 (m, 1H), 3.74–3.66 (m, 1H), 3.66–3.57 (m, 1H), 3.51 (p, J = 6.0 Hz, 1H), 3.24 (td,J=10.6,5.9Hz,2H), 2.82–2.69(m,1H), 2.48–2.38(m,1H), 2.39–2.29(m,1H), 2.04(dt,J=12.4,6.6Hz ,1H),1.16(s,3H),0.98(s,3H),0.95(s,3H),0.90(s,3H),0.87(s,3H),0.77(s,3H),0.76(s, 3H). ESI-MS: m/z 525.5 [M+H] + .
实施例52Example 52
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-3-羟基-吖丁啶(化合物A-96)N-(3β-hydroxy-oleanorane-13(18)-ene-28-acyl)-3-hydroxy-azetidine (compound A-96)
Figure PCTCN2020093248-appb-000088
Figure PCTCN2020093248-appb-000088
参照实施例2的方法，将N,N-二甲基乙二胺替换成氮杂环丁烷-3-醇，制得化合物A-96： 1H NMR(300MHz,CDCl 3)δ5.29–5.20(m,1H),3.90–3.72(m,1H),3.72–3.59(m,1H),3.58–3.40(m,2H),3.26–3.04(m,3H),2.13–1.99(m,2H),1.13(s,3H),0.98(s,3H),0.93(s,3H),0.89(s,6H),0.77(s,3H),0.71(s,3H).ESI-MS:m/z 510.5[M-H] -。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with azetidine-3-ol to obtain compound A-96: 1 H NMR (300MHz, CDCl 3 ) δ 5.29– 5.20(m,1H), 3.90--3.72(m,1H), 3.72--3.59(m,1H), 3.58--3.40(m,2H), 3.26--3.04(m,3H), 2.13--1.99(m,2H) ),1.13(s,3H),0.98(s,3H),0.93(s,3H),0.89(s,6H),0.77(s,3H),0.71(s,3H).ESI-MS:m/ z 510.5[MH] - .
实施例53Example 53
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-3-甲氧基-吖丁啶(化合物A-99)N-(3β-Hydroxy-oleanorane-13(18)-ene-28-acyl)-3-methoxy-azetidine (Compound A-99)
Figure PCTCN2020093248-appb-000089
Figure PCTCN2020093248-appb-000089
参照实施例2的方法，将N,N-二甲基乙二胺替换成3-甲氧基氮杂丁烷，制得化合物A-99： 1H NMR(300MHz,CDCl 3)δ4.56–4.17(m,2H),4.11(dt,J＝9.4,4.9Hz,1H),4.07–3.71(m,2H),3.29(s,3H),3.26–3.16(m,1H),2.80–2.68(m,1H),2.46–2.35(m,1H),2.17–2.08(m,1H),1.14(s,3H),0.98(s,3H),0.95(s,3H),0.91(s,3H),0.86(s,3H),0.76(s,3H),0.75(s,3H).ESI-MS:m/z 548.5[M+Na] +。 Referring to the method of Example 2, N,N-dimethylethylenediamine was replaced with 3-methoxyazetidine to obtain compound A-99: 1 H NMR (300MHz, CDCl 3 ) δ4.56– 4.17(m,2H),4.11(dt,J=9.4,4.9Hz,1H),4.07–3.71(m,2H), 3.29(s,3H), 3.26–3.16(m,1H), 2.80–2.68( m,1H), 2.46–2.35(m,1H), 2.17–2.08(m,1H), 1.14(s,3H), 0.98(s,3H), 0.95(s,3H), 0.91(s,3H) , 0.86 (s, 3H), 0.76 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 548.5 [M+Na] + .
实施例55Example 55
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-3-羧基-吖丁啶(化合物A-100)N-(3β-Hydroxy-oleanorane-13(18)-ene-28-acyl)-3-carboxy-azetidine (Compound A-100)
Figure PCTCN2020093248-appb-000090
Figure PCTCN2020093248-appb-000090
将化合物VIII-1(500mg，2.485mmol)溶于甲苯(7.5mL)中，依次加入1,8-二氮杂二环十一碳-7-烯(576mg，3.728mmol)和溴化苄(467mg，2.73mmol)，室温下搅拌反应4小时。TLC检测反应完全后，加入水(5mL)淬灭，乙酸乙酯(10mL×2)萃取，合并有机相，水(10mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝6：1)纯化，得化合物VIII-2(无色油状液体，296mg，产率40％)。Compound VIII-1 (500mg, 2.485mmol) was dissolved in toluene (7.5mL), and 1,8-diazabicycloundec-7-ene (576mg, 3.728mmol) and benzyl bromide (467mg , 2.73mmol), the reaction was stirred at room temperature for 4 hours. After the completion of the reaction detected by TLC, it was quenched by adding water (5mL), extracted with ethyl acetate (10mL×2), combined the organic phases, washed with water (10mL×2), dried with anhydrous sodium sulfate, filtered, the filtrate was concentrated, and passed through a silica gel column Purification by chromatography (petroleum ether: ethyl acetate = 6:1) gave compound VIII-2 (colorless oily liquid, 296 mg, yield 40%).
将化合物VIII-2(296mg，1.016mmol)溶于二氯甲烷(5mL)中，加入三氟乙酸(1mL)，室温搅拌反应。TLC检测反应完全后，减压蒸除溶剂，得化合物VIII-3，直接用于下一步。Compound VIII-2 (296 mg, 1.016 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added, and the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, the solvent was evaporated under reduced pressure to obtain compound VIII-3, which was directly used in the next step.
参照实施例36的方法，将D-脯氨酸苄酯盐酸盐替换成化合物VIII-3，制得化合物A-100： 1H NMR(300MHz,CDCl 3)δ4.65–4.01(m,4H),3.42–3.30(m,1H),3.24(dd,J＝10.3,5.5Hz,1H),2.85–2.65(m,1H),2.51–2.32(m,1H),2.26–2.04(m,1H),1.15(s,3H),0.98(s,3H),0.95(s,3H),0.92(s,3H),0.87(s,3H),0.76(s,3H),0.75(s,3H).ESI-MS:m/z 538.4[M-H] -。 According to the method of Example 36, D-proline benzyl ester hydrochloride was replaced with compound VIII-3 to prepare compound A-100: 1 H NMR (300MHz, CDCl 3 ) δ 4.65–4.01 (m, 4H ),3.42–3.30(m,1H), 3.24(dd,J=10.3,5.5Hz,1H), 2.85–2.65(m,1H), 2.51–2.32(m,1H), 2.26–2.04(m,1H) ), 1.15(s, 3H), 0.98(s, 3H), 0.95(s, 3H), 0.92(s, 3H), 0.87(s, 3H), 0.76(s, 3H), 0.75(s, 3H) .ESI-MS: m/z 538.4[MH] - .
实施例55Example 55
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基) 氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-25)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-(2-(2-acetamidoethoxyyl)benzoyl)oxy-oleanorane-13 (18)-ene-28-amide (Compound A-25)
Figure PCTCN2020093248-appb-000091
Figure PCTCN2020093248-appb-000091
参照实施例7的方法，将N-(2-氨基乙基)吗啉替换成4-(3-氨丙基)硫代吗啉-1,1-二氧化物，制得化合物A-25： 1H NMR(300MHz,CDCl 3)δ7.86–7.76(m,1H),7.70–7.63(m,1H),7.63–7.50(m,2H),6.58–6.46(m,1H),6.11–5.97(m,1H),4.77–4.66(m,1H),4.49–4.38(m,2H),3.69–3.59(m,2H),3.38–3.24(m,2H),3.18–2.94(m,6H),2.84(d,J＝15.3Hz,1H),2.68–2.53(m,2H),2.48(d,J＝12.8Hz,1H),2.43–2.34(m,1H),2.03(s,3H),1.21(s,3H),0.96(s,9H),0.93(s,3H),0.92(s,3H),0.75(s,3H).ESI-MS:m/z 862.5[M-H] -。 According to the method of Example 7, N-(2-aminoethyl)morpholine was replaced with 4-(3-aminopropyl)thiomorpholine-1,1-dioxide to obtain compound A-25: 1 H NMR (300MHz, CDCl 3 ) δ 7.86-7.76 (m, 1H), 7.70-7.63 (m, 1H), 7.63-7.50 (m, 2H), 6.58-6.46 (m, 1H), 6.11-5.97 (m,1H), 4.77-4.66(m,1H), 4.49-4.38(m,2H), 3.69-3.59(m,2H), 3.38-3.24(m,2H), 3.18-2.94(m,6H) ,2.84(d,J=15.3Hz,1H), 2.68–2.53(m,2H), 2.48(d,J=12.8Hz,1H), 2.43–2.34(m,1H),2.03(s,3H), 1.21 (s, 3H), 0.96 (s, 9H), 0.93 (s, 3H), 0.92 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 862.5 [MH] - .
实施例56Example 56
N-(3-(1-四氢吡咯基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-22)N-(3-(1-Tetrahydropyrrolyl)propyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide (Compound A-22 )
Figure PCTCN2020093248-appb-000092
Figure PCTCN2020093248-appb-000092
参照实施例13的方法，将3-二甲氨基丙胺替换成1-(3-氨基丙基)吡咯烷，制得化合物A-22： 1H NMR(300MHz,methanol-d 4)δ7.78(d,J＝7.4Hz,1H),7.69–7.54(m,2H),7.54–7.44(m,1H),4.82–4.72(m,1H),3.46–3.39(m,2H),3.18(t,J＝7.2Hz,2H),2.95(d,J＝14.6Hz,1H),2.62(d,J＝13.8Hz,1H),2.36–2.26(m,1H),2.14(s,3H),1.30(s,3H),1.06(s,6H),1.05(s,3H),1.03(s,3H),0.99(s,3H),0.84(s,2H).ESI-MS:m/z 713.6[M-H] -。 Referring to the method of Example 13, the 3-dimethylaminopropylamine was replaced with 1-(3-aminopropyl)pyrrolidine to obtain compound A-22: 1 H NMR (300MHz, methanol-d 4 )δ7.78( d,J=7.4Hz,1H), 7.69–7.54(m,2H), 7.54–7.44(m,1H), 4.82–4.72(m,1H), 3.46–3.39(m,2H), 3.18(t, J = 7.2Hz, 2H), 2.95 (d, J = 14.6 Hz, 1H), 2.62 (d, J = 13.8Hz, 1H), 2.36-2.26 (m, 1H), 2.14 (s, 3H), 1.30 ( s,3H),1.06(s,6H),1.05(s,3H),1.03(s,3H),0.99(s,3H),0.84(s,2H).ESI-MS:m/z 713.6(MH ] - .
实施例57Example 57
N-(3-(4-甲基-1-哌嗪基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-21)N-(3-(4-Methyl-1-piperazinyl)propyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-amide( Compound A-21)
Figure PCTCN2020093248-appb-000093
Figure PCTCN2020093248-appb-000093
参照实施例13的方法，将3-二甲胺丙胺替换成1-(3-氨丙基)-4-甲基哌嗪，制得化合物A-21： 1H NMR(300MHz,methanol-d 4)δ7.84–7.77(m,1H),7.74–7.68(m,1H),7.68–7.59(m,2H),4.78(dd,J＝11.0,4.2Hz,1H),3.82–3.43(m,8H),3.42–3.37(m,2H),3.20–3.09(m,2H),3.02(s,3H),2.95(d,J＝13.9Hz,1H), 2.62(d,J＝13.7Hz,1H),2.32(d,J＝13.3Hz,1H),1.30(s,3H),1.05(s,3H),1.04(s,3H),1.02(s,3H),0.98(s,3H),0.95(s,3H),0.83(s,3H).ESI-MS:m/z 742.6[M-H] -。 With reference to the method in Example 13, 3-dimethylaminopropylamine was replaced with 1-(3-aminopropyl)-4-methylpiperazine to obtain compound A-21: 1 H NMR (300MHz, methanol-d 4 )δ7.84–7.77(m,1H), 7.74–7.68(m,1H), 7.68–7.59(m,2H), 4.78(dd,J=11.0,4.2Hz,1H), 3.82–3.43(m, 8H),3.42–3.37(m,2H), 3.20–3.09(m,2H),3.02(s,3H), 2.95(d,J=13.9Hz,1H), 2.62(d,J=13.7Hz,1H ), 2.32(d,J=13.3Hz,1H),1.30(s,3H),1.05(s,3H),1.04(s,3H),1.02(s,3H),0.98(s,3H),0.95 (s, 3H), 0.83 (s, 3H). ESI-MS: m/z 742.6 [MH] - .
实施例58Example 58
N-(2-(1-哌嗪基)乙基)-3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-13)N-(2-(1-piperazinyl)ethyl)-3β-(2-(2-acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-13(18)-ene-28 -Amide (Compound A-13)
Figure PCTCN2020093248-appb-000094
Figure PCTCN2020093248-appb-000094
参照实施例7的方法，将N-(2-氨基乙基)吗啉替换成N-氨乙基哌嗪，制得化合物A-13： 1H NMR(300MHz,CDCl 3)δ7.81(d,J＝7.0Hz,1H),7.66(d,J＝7.0Hz,1H),7.62–7.51(m,2H),6.57–6.47(m,1H),6.19–6.11(m,1H),4.77–4.67(m,1H),4.49–4.41(m,2H),3.69–3.60(m,2H),3.49–3.28(m,2H),3.25–3.09(m,4H),2.90–2.71(m,5H),2.59–2.35(m,4H),2.03(s,3H),1.21(s,3H),0.97(s,3H),0.96(s,3H),0.95(s,3H),0.93(s,3H),0.90(s,3H),0.75(s,3H).ESI-MS:m/z801.6[M+H] +。 According to the method of Example 7, N-(2-aminoethyl)morpholine was replaced with N-aminoethylpiperazine to obtain compound A-13: 1 H NMR (300MHz, CDCl 3 )δ7.81(d ,J=7.0Hz,1H),7.66(d,J=7.0Hz,1H),7.62–7.51(m,2H),6.57–6.47(m,1H),6.19–6.11(m,1H),4.77– 4.67 (m, 1H), 4.49 - 4.41 (m, 2H), 3.69 - 3.60 (m, 2H), 3.49 - 3.28 (m, 2H), 3.25 - 3.09 (m, 4H), 2.90 - 2.71 (m, 5H) ), 2.59--2.35(m,4H),2.03(s,3H),1.21(s,3H),0.97(s,3H),0.96(s,3H),0.95(s,3H),0.93(s, 3H), 0.90 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 801.6 [M+H] + .
实施例59Example 59
3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸2-二甲氨基乙酯(化合物A-35)3β-(2-Carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acid 2-dimethylaminoethyl ester (Compound A-35)
Figure PCTCN2020093248-appb-000095
Figure PCTCN2020093248-appb-000095
取δ-齐墩果酸(δ-OA，2g，4.379mmol)溶于N,N-二甲基甲酰胺(20mL)中，依次加入1,2-二溴乙烷(3.75mL，43.79mmol)、碳酸钾(605mg，4.379mmol)和乙腈(2mL)，将反应液升温至50℃，搅拌反应3小时。TLC检测反应完全后，用乙酸乙酯(30mL)和水(10mL)处理反应液，水层用乙酸乙酯(20mL) 萃取，合并有机相，水(20mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝5：1)纯化，得化合物IX-1(白色固体，2.25g，产率91％)。Dissolve δ-oleanolic acid (δ-OA, 2g, 4.379mmol) in N,N-dimethylformamide (20mL), add 1,2-dibromoethane (3.75mL, 43.79mmol) in sequence , Potassium carbonate (605mg, 4.379mmol) and acetonitrile (2mL), the reaction solution was heated to 50°C, and the reaction was stirred for 3 hours. After the completion of the reaction detected by TLC, the reaction solution was treated with ethyl acetate (30mL) and water (10mL), the aqueous layer was extracted with ethyl acetate (20mL), the organic phases were combined, washed with water (20mL×2), and dried over anhydrous sodium sulfate , Filtration, concentration of the filtrate, and purification by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound IX-1 (white solid, 2.25 g, yield 91%).
取化合物IX-1(150mg，0.266mmol)溶于无水N,N-二甲基甲酰胺(8mL)中，依次加入二甲胺盐酸盐(33mg，0.405mmol)和碳酸钾(56mg，0.405mmol)，将反应液升温至50℃，搅拌反应。TLC检测反应完全后，用二氯甲烷(10mL)和水(5mL)处理反应液，水层用二氯甲烷(10mL×2)萃取，合并有机相，水(10mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝30：1)纯化，得化合物IX-2(白色固体，91mg，产率65％)。Take compound IX-1 (150mg, 0.266mmol) and dissolve it in anhydrous N,N-dimethylformamide (8mL), add dimethylamine hydrochloride (33mg, 0.405mmol) and potassium carbonate (56mg, 0.405 mmol), the reaction solution was heated to 50°C, and the reaction was stirred. After the completion of the reaction was detected by TLC, the reaction solution was treated with dichloromethane (10mL) and water (5mL), the aqueous layer was extracted with dichloromethane (10mL×2), the organic phases were combined, washed with water (10mL×2), and anhydrous sulfuric acid After drying with sodium, filtering, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain compound IX-2 (white solid, 91 mg, yield 65%).
将化合物IX-2(91mg，0.172mmol)溶于无水吡啶(8mL)中，加入邻苯二甲酸酐(255mg，1.72mmol)和4-二甲氨基吡啶(21mg，0.172mmol)，反应液升温至115℃搅拌过夜。TLC检测反应完全后，加入二氯甲烷(10mL)和水(10mL)处理反应液，水层用二氯甲烷(10mL)萃取，合并有机相，有机相依次用1N稀盐酸(10mL×3)和水(10mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝30：1)纯化，得化合物A-35(白色固体，83mg，产率72％)： 1H NMR(300MHz,CDCl 3)δ7.82(d,J＝7.2Hz,1H),7.58(d,J＝6.6Hz,1H),7.52–7.39(m,2H),4.83–4.74(m,1H),4.41(t,J＝5.5Hz,2H),3.11–2.95(m,2H),2.76(d,J＝14.6Hz,1H),2.61(s,6H),2.45(d,J＝13.7Hz,1H),2.17(d,J＝13.2Hz,1H),1.19(s,3H),1.02(s,3H),0.92(s,9H),0.89(s,3H),0.76(s,3H).ESI-MS:m/z 674.5[M-H] -。 Compound IX-2 (91 mg, 0.172 mmol) was dissolved in anhydrous pyridine (8 mL), phthalic anhydride (255 mg, 1.72 mmol) and 4-dimethylaminopyridine (21 mg, 0.172 mmol) were added, and the reaction solution was heated Stir at 115°C overnight. After the completion of the reaction was detected by TLC, dichloromethane (10mL) and water (10mL) were added to treat the reaction solution, the aqueous layer was extracted with dichloromethane (10mL), the organic phases were combined, and the organic phase was sequentially used with 1N dilute hydrochloric acid (10mL×3) and Wash with water (10mL×3), dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and purify by silica gel column chromatography (dichloromethane:methanol=30:1) to obtain compound A-35 (white solid, 83mg, yield 72%): 1 H NMR (300MHz, CDCl 3 ) δ 7.82 (d, J = 7.2 Hz, 1H), 7.58 (d, J = 6.6 Hz, 1H), 7.52–7.39 (m, 2H), 4.83– 4.74(m,1H), 4.41(t,J=5.5Hz,2H),3.11-2.95(m,2H),2.76(d,J=14.6Hz,1H), 2.61(s,6H), 2.45(d ,J=13.7Hz,1H),2.17(d,J=13.2Hz,1H),1.19(s,3H),1.02(s,3H),0.92(s,9H),0.89(s,3H),0.76 (s, 3H). ESI-MS: m/z 674.5 [MH] - .
实施例60Example 60
3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸2-(4-吗啉基)乙酯(化合物A-39)3β-(2-Carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acid 2-(4-morpholinyl) ethyl ester (Compound A-39)
Figure PCTCN2020093248-appb-000096
Figure PCTCN2020093248-appb-000096
参照实施例59的方法，将二甲胺盐酸盐替换成吗啉，制得化合物A-39： 1H NMR(300MHz,CDCl 3)δ7.92–7.81(m,1H),7.74–7.63(m,1H),7.58–7.46(m,2H),4.83–4.71(m,1H),4.28(t,J＝4.9Hz,2H),3.81–3.68(m,4H),2.81–2.66(m,3H),2.66–2.50(m,4H),2.42(d,J＝13.9Hz,1H),2.22–2.11(m,1H),1.17(s,3H),0.96(s,3H),0.90(s,12H),0.74(s,3H).ESI-MS:m/z 716.5[M-H] -。 According to the method of Example 59, dimethylamine hydrochloride was replaced with morpholine to obtain compound A-39: 1 H NMR (300MHz, CDCl 3 ) δ7.92-7.81(m,1H), 7.74-7.63( m,1H),7.58–7.46(m,2H), 4.83–4.71(m,1H), 4.28(t,J=4.9Hz,2H), 3.81–3.68(m,4H), 2.81–2.66(m, 3H), 2.66–2.50(m, 4H), 2.42(d, J=13.9Hz, 1H), 2.22–2.11(m, 1H), 1.17(s, 3H), 0.96(s, 3H), 0.90(s , 12H), 0.74 (s, 3H). ESI-MS: m/z 716.5 [MH] - .
实施例61Example 61
3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸2-(1-四氢吡咯基)乙酯(化合物A-40)3β-(2-Carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acid 2-(1-tetrahydropyrrolyl)ethyl ester (Compound A-40)
Figure PCTCN2020093248-appb-000097
Figure PCTCN2020093248-appb-000097
参照实施例59的方法，将二甲胺盐酸盐替换成四氢吡咯，制得化合物A-40： 1H NMR(300MHz,CDCl 3)δ7.79(d,J＝7.3Hz,1H),7.52(d,J＝6.6Hz,1H),7.47–7.34(m,2H),4.76(dd,J＝10.7,3.2Hz,1H),4.55–4.35(m,2H),3.31–3.05(m,6H),2.73(d,J＝12.0Hz,1H),2.42(d,J＝13.9Hz,1H),2.12(d,J＝13.1Hz,1H),2.08–1.96(m,4H),1.16(s,3H),1.02(s,3H),0.90(s,9H),0.86(s,3H),0.74(s,3H).ESI-MS:m/z 700.5[M-H] -。 According to the method of Example 59, dimethylamine hydrochloride was replaced with tetrahydropyrrole to obtain compound A-40: 1 H NMR (300MHz, CDCl 3 ) δ 7.79 (d, J = 7.3 Hz, 1H), 7.52(d,J=6.6Hz,1H),7.47–7.34(m,2H),4.76(dd,J＝10.7,3.2Hz,1H),4.55–4.35(m,2H),3.31–3.05(m, 6H), 2.73 (d, J = 12.0 Hz, 1H), 2.42 (d, J = 13.9 Hz, 1H), 2.12 (d, J = 13.1 Hz, 1H), 2.08-1.96 (m, 4H), 1.16 ( s, 3H), 1.02 (s, 3H), 0.90 (s, 9H), 0.86 (s, 3H), 0.74 (s, 3H). ESI-MS: m/z 700.5 [MH] - .
实施例62Example 62
3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸2-(1-哌啶基)乙酯(化合物A-38)3β-(2-Carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acid 2-(1-piperidinyl)ethyl ester (Compound A-38)
Figure PCTCN2020093248-appb-000098
Figure PCTCN2020093248-appb-000098
参照实施例59的方法，将二甲胺盐酸盐替换成哌啶，制得化合物A-38： 1H NMR(300MHz,methanol-d 4)δ7.79(d,J＝7.3Hz,1H),7.67(d,J＝7.2Hz,1H),7.59(t,J＝7.0Hz,1H),7.49(t,J＝7.2Hz,1H),4.81–4.73(m,1H),4.45–4.21(m,3H),2.95–2.84(m,1H),2.80(t,J＝5.8Hz,2H),2.74–2.62(m,4H),2.56(d,J＝14.6Hz,1H),2.31–2.22(m,1H),1.31(s,3H),1.07(s,6H),1.06(s,6H),1.02(s,3H),0.86(s,3H).ESI-MS:m/z 714.5[M-H] -。 According to the method of Example 59, the dimethylamine hydrochloride was replaced with piperidine to obtain compound A-38: 1 H NMR (300MHz, methanol-d 4 ) δ 7.79 (d, J = 7.3 Hz, 1H) ,7.67(d,J=7.2Hz,1H),7.59(t,J=7.0Hz,1H),7.49(t,J=7.2Hz,1H),4.81-4.73(m,1H),4.45-4.21( m, 3H), 2.95–2.84 (m, 1H), 2.80 (t, J = 5.8 Hz, 2H), 2.74–2.62 (m, 4H), 2.56 (d, J = 14.6 Hz, 1H), 2.31–2.22 (m,1H),1.31(s,3H),1.07(s,6H),1.06(s,6H),1.02(s,3H),0.86(s,3H).ESI-MS:m/z 714.5[ MH] - .
实施例63Example 63
3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸3-(1-四氢吡咯基)丙酯(化合物A-42)3β-(2-Carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acid 3-(1-tetrahydropyrrolyl)propyl ester (Compound A-42)
Figure PCTCN2020093248-appb-000099
Figure PCTCN2020093248-appb-000099
参照实施例59的方法，将1,2-二溴乙烷替换成1,3-二溴丙烷，二甲胺盐酸盐替换成四氢吡咯，制得化合物A-42： 1H NMR(300MHz,CDCl 3)δ7.74(d,J＝7.3Hz,1H),7.55(d,J＝7.0Hz,1H),7.46–7.31(m,2H),4.78–4.69(m,1H),4.24 –4.03(m,2H),3.19(s,4H),3.08–2.95(m,2H),2.74(d,J＝13.6Hz,1H),2.43(d,J＝14.1Hz,1H),2.26–2.01(m,7H),1.17(s,3H),0.99(s,3H),0.89(s,12H),0.74(s,3H).ESI-MS:m/z 714.5[M-H] -。 Referring to the method in Example 59, 1,2-dibromoethane was replaced with 1,3-dibromopropane, and dimethylamine hydrochloride was replaced with tetrahydropyrrole to obtain compound A-42: 1 H NMR (300MHz ,CDCl 3 )δ7.74(d,J=7.3Hz,1H),7.55(d,J=7.0Hz,1H),7.46–7.31(m,2H),4.78–4.69(m,1H),4.24 – 4.03 (m, 2H), 3.19 (s, 4H), 3.08–2.95 (m, 2H), 2.74 (d, J = 13.6 Hz, 1H), 2.43 (d, J = 14.1 Hz, 1H), 2.26–2.01 (m, 7H), 1.17 (s, 3H), 0.99 (s, 3H), 0.89 (s, 12H), 0.74 (s, 3H). ESI-MS: m/z 714.5 [MH] - .
实施例64Example 64
3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸3-二甲氨基丙酯(化合物A-43)3β-(2-Carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acid 3-dimethylaminopropyl ester (Compound A-43)
Figure PCTCN2020093248-appb-000100
Figure PCTCN2020093248-appb-000100
参照实施例59的方法，将1,2-二溴乙烷替换成1,3-二溴丙烷，制得化合物A-43： 1H NMR(300MHz,methanol-d 4)δ7.76(d,J＝7.5Hz,1H),7.64–7.53(m,2H),7.50–7.42(m,1H),4.81–4.69(m,1H),4.32–4.14(m,2H),3.18–3.08(m,2H),2.86(s,7H),2.55(d,J＝14.3Hz,1H),2.26–2.20(m,1H),2.20–2.08(m,2H),1.29(s,3H),1.04(s,6H),1.02(s,6H),0.98(s,3H),0.83(s,3H).ESI-MS:m/z 690.6[M+H] +。 Referring to the method in Example 59, 1,2-dibromoethane was replaced with 1,3-dibromopropane to obtain compound A-43: 1 H NMR (300MHz, methanol-d 4 )δ7.76(d, J = 7.5Hz, 1H), 7.64–7.53(m, 2H), 7.50–7.42(m, 1H), 4.81–4.69(m, 1H), 4.32–4.14(m, 2H), 3.18–3.08(m, 2H), 2.86 (s, 7H), 2.55 (d, J = 14.3 Hz, 1H), 2.26-2.20 (m, 1H), 2.20-2.08 (m, 2H), 1.29 (s, 3H), 1.04 (s , 6H), 1.02 (s, 6H), 0.98 (s, 3H), 0.83 (s, 3H). ESI-MS: m/z 690.6 [M+H] + .
实施例65Example 65
3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸2-(4-甲基-1-哌嗪基)乙酯(化合物A-48)3β-(2-(2-Acetylaminoethoxyacyl)benzoyl)oxy-oleanorane-13(18)-ene-28-acid 2-(4-methyl-1-piperazinyl) Ethyl ester (Compound A-48)
Figure PCTCN2020093248-appb-000101
Figure PCTCN2020093248-appb-000101
参照实施例7的方法制得化合物III-5。取化合物III-5(345mg，0.500mmol)溶于无水N,N-二甲基甲酰胺(8mL)中，依次加入1,2-二溴乙烷(435μL，5.00mmol)和碳酸钾(70mg，0.500mmol)，将反应液升温至50℃，搅拌反应3小时。TLC检测反应完全后，用乙酸乙酯(10mL)和水(10mL)处理反应液，水层用乙酸乙酯(10mL)萃取，合并有机相，水(10mL×2)洗，无水硫酸钠 干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝1：1)纯化，得化合物X-1(白色固体，348mg，产率87％)。According to the method of Example 7, compound III-5 was prepared. Dissolve compound III-5 (345mg, 0.500mmol) in anhydrous N,N-dimethylformamide (8mL), add 1,2-dibromoethane (435μL, 5.00mmol) and potassium carbonate (70mg , 0.500mmol), the reaction solution was heated to 50°C, and the reaction was stirred for 3 hours. After the completion of the reaction detected by TLC, the reaction solution was treated with ethyl acetate (10 mL) and water (10 mL), the aqueous layer was extracted with ethyl acetate (10 mL), the organic phases were combined, washed with water (10 mL×2), and dried over anhydrous sodium sulfate , Filtration, concentration of the filtrate, and purification by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound X-1 (white solid, 348 mg, yield 87%).
取化合物X-1(116mg，0.145mmol)溶于无水N,N-二甲基甲酰胺(8mL)中，依次加入N-甲基哌嗪(33μL，0.291mmol)和三乙胺(41μL，0.291mmol)，将反应液升温至50℃，搅拌反应。TLC检测反应完全后，用乙酸乙酯(10mL)和水(10mL)处理反应液，水层用乙酸乙酯(10mL)萃取，合并有机相，水(10mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝30：1)纯化，得化合物A-48(白色固体，63mg，产率53％)： 1H NMR(300MHz,CDCl 3)δ7.86–7.78(m,1H),7.69–7.63(m,1H),7.63–7.49(m,2H),6.62–6.50(m,1H),4.77–4.67(m,1H),4.45(t,J＝4.8Hz,2H),4.28–4.14(m,2H),3.70–3.59(m,2H),2.75(d,J＝13.0Hz,1H),2.68–2.33(m,11H),2.29(s,3H),2.21–2.15(m,1H),2.03(s,3H),1.17(s,3H),0.96(s,6H),0.95(s,3H),0.93(s,3H),0.92(s,3H),0.74(s,3H).ESI-MS:m/z 816.7[M+H] +。 Take compound X-1 (116mg, 0.145mmol) and dissolve it in anhydrous N,N-dimethylformamide (8mL), add N-methylpiperazine (33μL, 0.291mmol) and triethylamine (41μL, 0.291mmol), the reaction solution was heated to 50°C, and the reaction was stirred. After the completion of the reaction detected by TLC, the reaction solution was treated with ethyl acetate (10 mL) and water (10 mL), the aqueous layer was extracted with ethyl acetate (10 mL), the organic phases were combined, washed with water (10 mL×2), and dried over anhydrous sodium sulfate , Filter, concentrate the filtrate, and purify by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain compound A-48 (white solid, 63mg, yield 53%): 1 H NMR (300MHz, CDCl 3 ) δ7.86–7.78(m,1H), 7.69–7.63(m,1H), 7.63–7.49(m,2H), 6.62–6.50(m,1H), 4.77–4.67(m,1H), 4.45(t ,J=4.8Hz,2H), 4.28–4.14(m,2H), 3.70–3.59(m,2H), 2.75(d,J=13.0Hz,1H), 2.68–2.33(m,11H), 2.29( s,3H),2.21--2.15(m,1H),2.03(s,3H),1.17(s,3H),0.96(s,6H),0.95(s,3H),0.93(s,3H),0.92 (s, 3H), 0.74 (s, 3H). ESI-MS: m/z 816.7 [M+H] + .
实施例66Example 66
3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸2-(1-哌嗪基)乙酯(化合物A-47)3β-(2-(2-Acetylaminoethoxyacyl)benzoyl)oxy-oleanorane-13(18)-ene-28-acid 2-(1-piperazinyl)ethyl ester (Compound A -47)
Figure PCTCN2020093248-appb-000102
Figure PCTCN2020093248-appb-000102
参照实施例65的方法，将N-甲基哌嗪替换成哌嗪，制得化合物A-47： 1H NMR(300MHz,CDCl 3)δ7.84–7.79(m,1H),7.69–7.63(m,1H),7.62–7.50(m,2H),6.61–6.50(m,1H),4.77–4.67(m,1H),4.44(t,J＝5.0Hz,2H),4.26–4.12(m,2H),3.71–3.60(m,2H),3.25–3.13(m,4H),2.91–2.70(m,5H),2.66(t,J＝5.5Hz,2H),2.42(d,J＝14.1Hz,1H),2.21–2.11(m,1H),2.03(s,3H),1.18(s,3H),0.97(s,3H),0.96(s,3H),0.95(s,3H),0.91(s,6H),0.74(s,3H).ESI-MS:m/z 802.7[M+H] +。 According to the method of Example 65, N-methylpiperazine was replaced with piperazine to obtain compound A-47: 1 H NMR (300MHz, CDCl 3 )δ7.84-7.79(m,1H), 7.69-7.63( m,1H), 7.62–7.50(m,2H), 6.61–6.50(m,1H), 4.77–4.67(m,1H), 4.44(t,J=5.0Hz,2H), 4.26–4.12(m, 2H), 3.71–3.60(m,2H), 3.25–3.13(m,4H), 2.91–2.70(m,5H), 2.66(t,J=5.5Hz,2H),2.42(d,J=14.1Hz ,1H),2.21--2.11(m,1H),2.03(s,3H),1.18(s,3H),0.97(s,3H),0.96(s,3H),0.95(s,3H),0.91( s, 6H), 0.74 (s, 3H). ESI-MS: m/z 802.7 [M+H] + .
实施例67Example 67
N-(3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰基)-吖丁啶(化合物A-106)N-(3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acyl)-azetidine (compound A-106)
Figure PCTCN2020093248-appb-000103
Figure PCTCN2020093248-appb-000103
参照实施例2的方法，将N,N-二甲基乙二胺替换成杂氮环丁烷，制得化合 物A-106： 1H NMR(300MHz,CDCl 3)δ8.00–7.36(m,4H),4.88–4.64(m,1H),4.34–3.93(m,4H),2.83–2.66(m,1H),2.48–2.33(m,1H),1.15(s,3H),0.93(s,9H),0.85(s,6H),0.77(s,3H).ESI-MS:m/z 642.5[M-H] -。 Referring to the method of Example 2, N,N-dimethylethylenediamine was replaced with azacyclobutane to obtain compound A-106: 1 H NMR (300MHz, CDCl 3 )δ8.00-7.36(m, 4H), 4.88–4.64(m,1H), 4.34–3.93(m,4H), 2.83–2.66(m,1H), 2.48–2.33(m,1H), 1.15(s,3H), 0.93(s, 9H), 0.85 (s, 6H), 0.77 (s, 3H). ESI-MS: m/z 642.5 [MH] - .
实施例68Example 68
N-(3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酰基)-3-甲氧基-吖丁啶(化合物A-107)N-(3β-(2-carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acyl)-3-methoxy-azetidine (Compound A-107)
Figure PCTCN2020093248-appb-000104
Figure PCTCN2020093248-appb-000104
参照实施例2的方法，将N,N-二甲基乙二胺替换成3-甲氧基氮杂丁烷，制得化合物A-107： 1H NMR(300MHz,DMSO-d 6)δ13.24(s,1H),7.76–7.68(m,1H),7.66–7.57(m,3H),4.71–4.56(m,1H),4.50–3.58(m,5H),3.19(s,3H),2.79–2.63(m,1H),2.34(d,J＝13.5Hz,1H),2.10–1.97(m,1H),1.14(s,3H),0.91(s,3H),0.88(s,9H),0.82(s,3H),0.73(s,3H).ESI-MS:m/z 672.5[M-H] -。 With reference to the method of Example 2, N,N-dimethylethylenediamine was replaced with 3-methoxyazetidine to obtain compound A-107: 1 H NMR (300MHz, DMSO-d 6 )δ13. 24 (s, 1H), 7.76-7.68 (m, 1H), 7.66-7.57 (m, 3H), 4.71-4.56 (m, 1H), 4.50-3.58 (m, 5H), 3.19 (s, 3H), 2.79–2.63(m,1H), 2.34(d,J=13.5Hz,1H), 2.10–1.97(m,1H), 1.14(s,3H), 0.91(s,3H), 0.88(s,9H) , 0.82(s, 3H), 0.73(s, 3H). ESI-MS: m/z 672.5[MH] - .
实施例69Example 69
N-(2-(1-哌嗪基)乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-72)N-(2-(1-piperazinyl)ethyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide (compound A-72)
Figure PCTCN2020093248-appb-000105
Figure PCTCN2020093248-appb-000105
参照实施例13的方法，将3-二甲氨基丙胺替换成N-氨乙基哌嗪，制得化合物A-72： 1H NMR(300MHz,DMSO-d 6)δ9.77–9.47(m,2H),7.71–7.58(m,1H),5.30–5.19(m,1H),3.97–3.58(m,4H),3.52–3.32(m,6H),3.14–2.95(m,3H),2.88–2.76(m,1H),1.09(s,3H),0.89(s,6H),0.87(s,3H),0.85(s,3H),0.67(s,3H),0.66(s,3H).ESI-MS:m/z 568.5[M+H] +。 According to the method of Example 13, the 3-dimethylaminopropylamine was replaced with N-aminoethylpiperazine to obtain compound A-72: 1 H NMR (300MHz, DMSO-d 6 )δ9.77–9.47(m, 2H), 7.71–7.58(m,1H), 5.30–5.19(m,1H), 3.97–3.58(m,4H), 3.52–3.32(m,6H), 3.14–2.95(m,3H), 2.88– 2.76(m,1H),1.09(s,3H),0.89(s,6H),0.87(s,3H),0.85(s,3H),0.67(s,3H),0.66(s,3H).ESI -MS: m/z 568.5 [M+H] + .
实施例70Example 70
3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸2-三甲基铵基乙酯溴化物(化合物A-45)3β-(2-(2-Acetylaminoethoxyyl)benzoyl)oxy-oleanorane-13(18)-ene-28-acid 2-trimethylammonium ethyl bromide (Compound A -45)
Figure PCTCN2020093248-appb-000106
Figure PCTCN2020093248-appb-000106
参照实施例65的方法，将N-甲基哌嗪替换成三甲胺，制得化合物A-45： 1H NMR(300MHz,methanol-d 4)δ7.78–7.71(m,2H),7.66–7.59(m,2H),4.76–4.67(m,2H),4.61–4.51(m,2H),4.36(t,J＝5.5Hz,2H),3.73(t,J＝5.0Hz,2H), 3.53(t,J＝5.4Hz,2H),3.24(s,9H),2.81(d,J＝15.0Hz,1H),2.49(d,J＝13.9Hz,1H),2.19(d,J＝13.5Hz,1H),1.96(s,4H),1.24(s,3H),0.99(s,3H),0.97(s,3H),0.97(s,3H),0.94(s,3H),0.92(s,3H),0.79(s,3H).ESI-MS:m/z 775.6[M-Br] +。 According to the method of Example 65, N-methylpiperazine was replaced with trimethylamine to obtain compound A-45: 1 H NMR (300MHz, methanol-d 4 )δ7.78–7.71(m,2H),7.66– 7.59(m,2H), 4.76–4.67(m,2H), 4.61–4.51(m,2H), 4.36(t,J=5.5Hz,2H), 3.73(t,J=5.0Hz,2H), 3.53 (t,J=5.4Hz,2H), 3.24(s,9H), 2.81(d,J=15.0Hz,1H), 2.49(d,J=13.9Hz,1H), 2.19(d,J=13.5Hz ,1H),1.96(s,4H),1.24(s,3H),0.99(s,3H),0.97(s,3H),0.97(s,3H),0.94(s,3H),0.92(s, 3H), 0.79 (s, 3H). ESI-MS: m/z 775.6 [M-Br] + .
实施例71Example 71
3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸3-三甲基铵基丙酯溴化物(化合物A-46)3β-(2-(2-Acetylaminoethoxyacyl)benzoyl)oxy-oleanorane-13(18)-ene-28-acid 3-trimethylammonium propyl bromide (Compound A -46)
Figure PCTCN2020093248-appb-000107
Figure PCTCN2020093248-appb-000107
参照实施例65的方法，将1,2-二溴乙烷替换成1,3-二溴丙烷，N-甲基哌嗪替换成三甲胺，制得化合物A-46： 1H NMR(300MHz,methanol-d 4)δ7.83–7.76(m,2H),7.71–7.64(m,2H),4.81–4.73(m,1H),4.41(t,J＝5.5Hz,2H),4.23(dd,J＝11.7,5.9Hz,2H),3.58(t,J＝5.5Hz,2H),3.48(dd,J＝10.2,6.5Hz,2H),3.23(s,9H),2.86(d,J＝14.4Hz,1H),2.56(d,J＝14.1Hz,1H),2.30–2.16(m,3H),2.01(s,4H),1.29(s,3H),1.04(s,3H),1.02(s,6H),0.98(s,3H),0.97(s,3H),0.83(s,3H).ESI-MS:m/z 789.6[M-Br] +。 Referring to the method in Example 65, 1,2-dibromoethane was replaced with 1,3-dibromopropane, and N-methylpiperazine was replaced with trimethylamine to obtain compound A-46: 1 H NMR (300MHz, methanol-d 4 )δ7.83–7.76(m,2H), 7.71–7.64(m,2H), 4.81–4.73(m,1H), 4.41(t,J=5.5Hz,2H), 4.23(dd, J = 11.7, 5.9 Hz, 2H), 3.58 (t, J = 5.5 Hz, 2H), 3.48 (dd, J = 10.2, 6.5 Hz, 2H), 3.23 (s, 9H), 2.86 (d, J = 14.4 Hz,1H),2.56(d,J=14.1Hz,1H),2.30–2.16(m,3H),2.01(s,4H),1.29(s,3H),1.04(s,3H),1.02(s , 6H), 0.98 (s, 3H), 0.97 (s, 3H), 0.83 (s, 3H). ESI-MS: m/z 789.6 [M-Br] + .
实施例72Example 72
3β-(2-羧基苯甲酰基)氧基-齐墩果烷-13(18)-烯-28-酸2-(4-甲基-1-哌嗪基)乙酯(化合物A-37)3β-(2-Carboxybenzoyl)oxy-oleanorane-13(18)-ene-28-acid 2-(4-methyl-1-piperazinyl)ethyl ester (Compound A-37)
Figure PCTCN2020093248-appb-000108
Figure PCTCN2020093248-appb-000108
参照实施例59的方法，将二甲胺盐酸盐替换成N-甲基哌嗪，制得化合物A-37： 1H NMR(300MHz,methanol-d 4)δ7.84–7.76(m,1H),7.75–7.68(m,1H),7.68–7.60(m,2H),4.77(dd,J＝10.6,5.2Hz,1H),4.52–4.40(m,2H),3.80–3.25(m,10H),3.02(s,3H),2.85(d,J＝14.8Hz,1H),2.53(d,J＝14.2Hz,1H),2.30–2.22(m,1H),1.28(s,3H),1.03(s,3H),1.02(s,3H),1.01(s,3H),0.99(s,3H),0.95(s,3H),0.83(s,3H).ESI-MS:m/z 731.6[M+H] +。 According to the method of Example 59, the dimethylamine hydrochloride was replaced with N-methylpiperazine to obtain compound A-37: 1 H NMR (300MHz, methanol-d 4 )δ7.84–7.76 (m, 1H ), 7.75–7.68(m,1H), 7.68–7.60(m,2H), 4.77(dd,J＝10.6,5.2Hz,1H),4.52–4.40(m,2H),3.80–3.25(m,10H ), 3.02 (s, 3H), 2.85 (d, J = 14.8 Hz, 1H), 2.53 (d, J = 14.2 Hz, 1H), 2.30-2.22 (m, 1H), 1.28 (s, 3H), 1.03 (s,3H),1.02(s,3H),1.01(s,3H),0.99(s,3H),0.95(s,3H),0.83(s,3H).ESI-MS:m/z 731.6[ M+H] + .
实施例73Example 73
N-(2-(二甲氨基)乙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-1)N-(2-(Dimethylamino)ethyl)-3β-hydroxy-oleanane-12-ene-28-amide (compound B-1)
Figure PCTCN2020093248-appb-000109
Figure PCTCN2020093248-appb-000109
Figure PCTCN2020093248-appb-000110
Figure PCTCN2020093248-appb-000110
取齐墩果酸(OA，5g，0.011mol)溶于无水吡啶(35mL)中，搅拌下缓慢滴加醋酸酐(4mL)，滴加完毕后加热回流反应1小时。TLC检测反应完全后，反应液冷却至室温，将其滴入冰水(20mL×2)中，析出大量白色固体，抽滤，滤饼用水洗(10mL×3)，正己烷洗(10mL×3)，真空干燥，乙醇重结晶，得化合物XI-1(白色固体，4.52g，产率83％)。Take oleanolic acid (OA, 5g, 0.011mol) and dissolve it in anhydrous pyridine (35mL), slowly add acetic anhydride (4mL) dropwise with stirring, and heat and reflux for 1 hour after the addition is complete. After the completion of the reaction detected by TLC, the reaction solution was cooled to room temperature and dropped into ice water (20mL×2). A large amount of white solid was precipitated. The filter cake was washed with water (10mL×3) and n-hexane (10mL×3). ), vacuum drying, and ethanol recrystallization to obtain compound XI-1 (white solid, 4.52 g, yield 83%).
取化合物XI-1(150mg，0.301mmol)溶于无水二氯甲烷(8mL)中，搅拌下缓慢滴加草酰氯(130μL，1.504mmol)和N,N-二甲基甲酰胺(1滴)，室温反应3小时。TLC检测反应完全后，减压蒸除溶剂，得化合物XI-2(黄色固体，155mg，产率100％)。Dissolve compound XI-1 (150mg, 0.301mmol) in anhydrous dichloromethane (8mL), add oxalyl chloride (130μL, 1.504mmol) and N,N-dimethylformamide (1 drop) slowly and dropwise with stirring , React at room temperature for 3 hours. After the completion of the reaction detected by TLC, the solvent was evaporated under reduced pressure to obtain compound XI-2 (yellow solid, 155 mg, yield 100%).
将N,N-二甲基乙二胺(66μL，0.602mmol)溶于无水二氯甲烷(5mL)中，搅拌下缓慢滴加化合物XI-2(155mg，0.301mmol)的无水二氯甲烷(5mL)溶液和三乙胺(84μL，0.602mmol)，室温反应5小时。TLC检测反应完全后，加入二氯甲烷(10mL)稀释反应液，反应液依次用饱和碳酸氢钠溶液(10mL×3)、水(10mL×3)和饱和食盐水(10mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝100：1～50：1)纯化，得化合物XI-3(白色固体，156mg，产率91％)。N,N-Dimethylethylenediamine (66μL, 0.602mmol) was dissolved in dry dichloromethane (5mL), and compound XI-2 (155mg, 0.301mmol) in dry dichloromethane was slowly added dropwise with stirring (5mL) The solution and triethylamine (84μL, 0.602mmol) were reacted at room temperature for 5 hours. After the completion of the reaction detected by TLC, dichloromethane (10mL) was added to dilute the reaction solution, and the reaction solution was washed with saturated sodium bicarbonate solution (10mL×3), water (10mL×3) and saturated brine (10mL×3) successively. Drying with water sodium sulfate, filtering, concentrating the filtrate, and purifying by silica gel column chromatography (dichloromethane: methanol = 100:1-50:1) to obtain compound XI-3 (white solid, 156mg, yield 91%).
将化合物XI-3(156mg，0.274mmol)悬浮于甲醇(10mL)中，加入氢氧化钾(154mg，2.74mmol)，将反应液升温至65℃，反应5小时。TLC检测反应完全后，反应液冷却至室温，减压蒸除溶剂，残余物用乙酸乙酯(20mL)和水(20mL)处理，有机相依次用水(20mL×3)和饱和食盐水(20mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝10：1)纯化，得化合物B-1(白色固体，123mg，产率85％)： 1H NMR(300MHz,CDCl 3)δ6.59–6.45(m,1H),5.35(t,J＝3.4Hz,1H),3.44–3.29(m,1H),3.27–3.07(m,2H),2.59–2.46(m,1H),2.37(t,J＝6.1Hz,2H),2.23(s,6H),1.16(s,3H),0.99(s,3H),0.91(s,9H),0.79(s,3H),0.78(s,3H).ESI-MS:m/z 527.5[M+H] +。 Compound XI-3 (156 mg, 0.274 mmol) was suspended in methanol (10 mL), potassium hydroxide (154 mg, 2.74 mmol) was added, the reaction solution was heated to 65° C. and reacted for 5 hours. After the completion of the reaction detected by TLC, the reaction solution was cooled to room temperature, and the solvent was evaporated under reduced pressure. The residue was treated with ethyl acetate (20mL) and water (20mL). The organic phase was sequentially water (20mL×3) and saturated brine (20mL× 3) Wash, dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and purify by silica gel column chromatography (dichloromethane: methanol = 10:1) to obtain compound B-1 (white solid, 123 mg, yield 85%): 1 H NMR(300MHz, CDCl 3 )δ6.59–6.45(m,1H), 5.35(t,J=3.4Hz,1H), 3.44–3.29(m,1H), 3.27–3.07(m,2H), 2.59–2.46(m,1H), 2.37(t,J=6.1Hz,2H), 2.23(s,6H), 1.16(s,3H),0.99(s,3H),0.91(s,9H),0.79 (s, 3H), 0.78 (s, 3H). ESI-MS: m/z 527.5 [M+H] + .
实施例74Example 74
N-(3β-羟基-齐墩果烷-12-烯-28-酰胺)-吗啉(化合物B-2)N-(3β-Hydroxy-oleanan-12-en-28-amide)-morpholine (Compound B-2)
Figure PCTCN2020093248-appb-000111
Figure PCTCN2020093248-appb-000111
参照实施例73的方法，将N,N-二甲基乙二胺替换成吗啉，制得化合物B-2： 1H NMR(300MHz,CDCl 3)δ5.33–5.21(m,1H),3.76–3.54(m,8H),3.28–3.15(m,1H),3.15–3.03(m,1H),1.16(s,3H),1.00(s,3H),0.95(s,3H),0.92(s,6H),0.80(s,3H),0.75(s,3H).ESI-MS:m/z 526.5[M+H] +。 Referring to the method of Example 73, N,N-dimethylethylenediamine was replaced with morpholine to obtain compound B-2: 1 H NMR (300MHz, CDCl 3 )δ5.33-5.21(m,1H), 3.76–3.54(m,8H), 3.28–3.15(m,1H), 3.15–3.03(m,1H), 1.16(s,3H), 1.00(s,3H), 0.95(s,3H), 0.92( s, 6H), 0.80 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 526.5 [M+H] + .
实施例75Example 75
N-(2-(二甲氨基)乙基)-N-甲基-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-3)N-(2-(Dimethylamino)ethyl)-N-methyl-3β-hydroxy-oleanane-12-ene-28-amide (compound B-3)
Figure PCTCN2020093248-appb-000112
Figure PCTCN2020093248-appb-000112
参照实施例73的方法，将N,N-二甲基乙二胺替换成N,N,N'-三甲基乙二胺，制得化合物B-3： 1H NMR(300MHz,CDCl 3)δ5.26(t,J＝3.3Hz,1H),3.43(t,J＝7.2Hz,2H),3.26–3.17(m,1H),3.16–3.12(m,1H),3.10(s,3H),2.54–2.31(m,3H),2.25(s,6H),1.14(s,3H),0.99(s,3H),0.93(s,3H),0.91(s,3H),0.89(s,3H),0.78(s,3H),0.75(s,3H).ESI-MS:m/z 541.6[M+H] +。 Referring to the method of Example 73, N,N-dimethylethylenediamine was replaced with N,N,N'-trimethylethylenediamine to obtain compound B-3: 1 H NMR (300MHz, CDCl 3 ) δ5.26(t,J=3.3Hz,1H), 3.43(t,J=7.2Hz,2H), 3.26–3.17(m,1H), 3.16–3.12(m,1H), 3.10(s,3H) ,2.54--2.31(m,3H),2.25(s,6H),1.14(s,3H),0.99(s,3H),0.93(s,3H),0.91(s,3H),0.89(s,3H) ), 0.78 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 541.6 [M+H] + .
实施例76Example 76
N-(3-(二甲氨基)丙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-4)N-(3-(dimethylamino)propyl)-3β-hydroxy-oleanorane-12-en-28-amide (compound B-4)
Figure PCTCN2020093248-appb-000113
Figure PCTCN2020093248-appb-000113
参照实施例73的方法，将N,N-二甲基乙二胺替换成3-二甲氨基丙胺，制得化合物B-4： 1H NMR(300MHz,CDCl 3)δ7.03–6.96(m,1H),5.33–5.28(m,1H),3.59–3.45(m,1H),3.27–3.15(m,1H),3.08–2.94(m,1H),2.60–2.47(m,1H),2.47–2.26(m,2H),2.23(s,6H),1.15(s,3H),0.99(s,3H),0.91(s,9H),0.78(s,3H),0.76(s,3H).ESI-MS:m/z 563.5[M+Na] +。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with 3-dimethylaminopropylamine to obtain compound B-4: 1 H NMR (300MHz, CDCl 3 )δ7.03–6.96 (m ,1H),5.33–5.28(m,1H),3.59–3.45(m,1H), 3.27–3.15(m,1H),3.08–2.94(m,1H), 2.60–2.47(m,1H), 2.47 --2.26 (m, 2H), 2.23 (s, 6H), 1.15 (s, 3H), 0.99 (s, 3H), 0.91 (s, 9H), 0.78 (s, 3H), 0.76 (s, 3H). ESI-MS: m/z 563.5 [M+Na] + .
实施例77Example 77
N-(2-(二乙氨基)乙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-5)N-(2-(Diethylamino)ethyl)-3β-hydroxy-oleanane-12-ene-28-amide (compound B-5)
Figure PCTCN2020093248-appb-000114
Figure PCTCN2020093248-appb-000114
参照实施例73的方法，将N,N-二甲基乙二胺替换成N,N-二乙基乙二胺，制得化合物B-5： 1H NMR(300MHz,CDCl 3)δ6.63–6.53(m,1H),5.38–5.33(m,1H),3.53–3.41(m,1H),3.28–3.17(m,1H),3.10–2.97(m,1H),2.65–2.39(m,7H),1.17(s,3H),1.04(t,J＝7.1Hz,6H),1.01(s,3H),0.92(s,9H),0.80(s,3H),0.78(s,3H).ESI-MS:m/z 577.5[M+Na] +。 Referring to the method of Example 73, N,N-dimethylethylenediamine was replaced with N,N-diethylethylenediamine to obtain compound B-5: 1 H NMR (300MHz, CDCl 3 ) δ6.63 --6.53(m,1H),5.38–5.33(m,1H),3.53–3.41(m,1H), 3.28–3.17(m,1H), 3.10–2.97(m,1H), 2.65–2.39(m, 7H), 1.17 (s, 3H), 1.04 (t, J = 7.1 Hz, 6H), 1.01 (s, 3H), 0.92 (s, 9H), 0.80 (s, 3H), 0.78 (s, 3H). ESI-MS: m/z 577.5 [M+Na] + .
实施例78Example 78
N-(2-(1-哌啶基)乙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-6)N-(2-(1-piperidinyl)ethyl)-3β-hydroxy-oleanorane-12-en-28-amide (compound B-6)
Figure PCTCN2020093248-appb-000115
Figure PCTCN2020093248-appb-000115
参照实施例73的方法，将N,N-二甲基乙二胺替换成1-(2-氨乙基)哌啶，制得化合物B-6： 1H NMR(300MHz,CDCl 3)δ6.65–6.55(m,1H),5.43–5.28(m,1H),3.45–3.32(m,1H),3.27–3.08(m,2H),2.57–2.47(m,1H),2.47–2.24(m,6H),1.16(s,3H),0.99(s,3H),0.91(s,9H),0.78(s,3H),0.77(s,3H).ESI-MS:m/z567.5[M+H] +。 Referring to the method of Example 73, N,N-dimethylethylenediamine was replaced with 1-(2-aminoethyl)piperidine to obtain compound B-6: 1 H NMR (300MHz, CDCl 3 )δ6. 65-6.55(m, 1H), 5.43-5.28(m, 1H), 3.45-3.32(m, 1H), 3.27-3.08(m, 2H), 2.57-2.47(m, 1H), 2.47-2.24(m ,6H),1.16(s,3H),0.99(s,3H),0.91(s,9H),0.78(s,3H),0.77(s,3H).ESI-MS:m/z567.5(M +H] + .
实施例79Example 79
N-(2-(4-吗啉烷基)乙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-7)N-(2-(4-morpholinoalkyl)ethyl)-3β-hydroxy-oleanorane-12-en-28-amide (compound B-7)
Figure PCTCN2020093248-appb-000116
Figure PCTCN2020093248-appb-000116
参照实施例73的方法，将N,N-二甲基乙二胺替换成N-(2-氨基乙基)吗啉，制得化合物B-7： 1H NMR(300MHz,CDCl 3)δ6.54–6.42(m,1H),5.42–5.30(m,1H),3.78–3.62(m,4H),3.50–3.33(m,1H),3.30–3.08(m,2H),2.58–2.49(m,1H),2.49–2.32(m,6H),1.16(s,3H),0.99(s,3H),0.91(s,6H),0.89(s,3H),0.78(s,3H),0.76(s,3H).ESI-MS:m/z 569.5[M+H] +。 Referring to the method in Example 73, N,N-dimethylethylenediamine was replaced with N-(2-aminoethyl)morpholine to obtain compound B-7: 1 H NMR (300MHz, CDCl 3 )δ6. 54–6.42(m,1H), 5.42–5.30(m,1H), 3.78–3.62(m,4H), 3.50–3.33(m,1H), 3.30–3.08(m,2H), 2.58–2.49(m ,1H),2.49--2.32(m,6H),1.16(s,3H),0.99(s,3H),0.91(s,6H),0.89(s,3H),0.78(s,3H),0.76( s, 3H). ESI-MS: m/z 569.5 [M+H] + .
实施例80Example 80
N-(3-(4-吗啉烷基)丙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-8)N-(3-(4-morpholinoalkyl)propyl)-3β-hydroxy-oleanorane-12-en-28-amide (compound B-8)
Figure PCTCN2020093248-appb-000117
Figure PCTCN2020093248-appb-000117
参照实施例73的方法，将N,N-二甲基乙二胺替换成N-(3-氨基丙基)吗啉，制得化合物B-8： 1H NMR(300MHz,CDCl 3)δ6.30–6.21(m,1H),5.40–5.31(m,1H),3.79–3.66(m,4H),3.51–3.36(m,1H),3.27–3.16(m,1H),3.13–2.98(m,1H),2.61–2.50(m,1H),2.50–2.31(m,6H),1.16(s,3H),0.99(s,3H),0.90(s,9H),0.78(s,3H),0.76(s,3H).ESI-MS:m/z 569.5[M+H] +。 Referring to the method of Example 73, N,N-dimethylethylenediamine was replaced with N-(3-aminopropyl)morpholine to obtain compound B-8: 1 H NMR (300MHz, CDCl 3 )δ6. 30--6.21(m,1H), 5.40--5.31(m,1H), 3.79--3.66(m,4H), 3.51--3.36(m,1H), 3.27--3.16(m,1H), 3.13--2.98(m ,1H), 2.61–2.50(m,1H), 2.50–2.31(m,6H), 1.16(s,3H), 0.99(s,3H), 0.90(s,9H), 0.78(s,3H), 0.76(s, 3H). ESI-MS: m/z 569.5 [M+H] + .
实施例81Example 81
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-硫代吗啉-1,1-二氧化物(化合物B-9)N-(3β-hydroxy-oleanane-12-ene-28-acyl)-thiomorpholine-1,1-dioxide (compound B-9)
Figure PCTCN2020093248-appb-000118
Figure PCTCN2020093248-appb-000118
参照实施例73的方法，将N,N-二甲基乙二胺替换成硫代吗啉-1,1-二氧化物，制得化合物B-9： 1H NMR(300MHz,CDCl 3)δ5.32–5.23(m,1H),4.09(t,J＝5.2Hz,4H),3.26–3.14(m,1H),3.11–2.90(m,4H),2.29–2.10(m,1H),1.15(s,3H),0.99(s,3H),0.93(s,3H),0.91(s,3H),0.90(s,3H),0.78(s,3H),0.71(s,3H).ESI-MS:m/z 574.4[M+H] +。 Referring to the method of Example 73, N,N-dimethylethylenediamine was replaced with thiomorpholine-1,1-dioxide to obtain compound B-9: 1 H NMR (300MHz, CDCl 3 )δ5 .32–5.23(m,1H),4.09(t,J=5.2Hz,4H), 3.26–3.14(m,1H), 3.11–2.90(m,4H), 2.29–2.10(m,1H), 1.15 (s,3H),0.99(s,3H),0.93(s,3H),0.91(s,3H),0.90(s,3H),0.78(s,3H),0.71(s,3H).ESI- MS: m/z 574.4 [M+H] + .
实施例82Example 82
N-(2-(1-四氢吡咯基)乙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-10)N-(2-(1-Tetrahydropyrrolyl)ethyl)-3β-hydroxy-oleanolane-12-en-28-amide (compound B-10)
Figure PCTCN2020093248-appb-000119
Figure PCTCN2020093248-appb-000119
参照实施例73的方法，将N,N-二甲基乙二胺替换成1-(2-氨乙基)吡咯烷，制得化合物B-10： 1H NMR(300MHz,CDCl 3)δ6.64–6.52(m,1H),5.37–5.26(m,1H),3.50–3.34(m,1H),3.19(d,J＝4.7Hz,1H),2.63–2.44(m,4H),1.14(s,2H),0.97(s,2H),0.89(s,5H),0.76(s,3H).ESI-MS:m/z 553.5[M+H] +。 Referring to the method of Example 73, N,N-dimethylethylenediamine was replaced with 1-(2-aminoethyl)pyrrolidine to obtain compound B-10: 1 H NMR (300MHz, CDCl 3 )δ6. 64–6.52(m,1H), 5.37–5.26(m,1H), 3.50–3.34(m,1H), 3.19(d,J=4.7Hz,1H), 2.63–2.44(m,4H), 1.14( s, 2H), 0.97 (s, 2H), 0.89 (s, 5H), 0.76 (s, 3H). ESI-MS: m/z 553.5 [M+H] + .
实施例83Example 83
N-(2-(1-哌嗪基)乙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-11)N-(2-(1-piperazinyl)ethyl)-3β-hydroxy-oleanorane-12-en-28-amide (compound B-11)
Figure PCTCN2020093248-appb-000120
Figure PCTCN2020093248-appb-000120
参照实施例73的方法，将N,N-二甲基乙二胺替换成N-氨乙基哌嗪，制得化合物B-11： 1H NMR(300MHz,DMSO-d 6)δ9.77–9.47(m,2H),7.71–7.58(m,1H),5.30–5.19(m,1H),3.97–3.58(m,4H),3.52–3.32(m,6H),3.14–2.95(m,3H),2.88–2.76(m,1H),1.09(s,3H),0.89(s,6H),0.87(s,3H),0.85(s,3H),0.67(s,3H),0.66(s,3H).ESI-MS:m/z 568.5[M+H] +。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with N-aminoethylpiperazine to obtain compound B-11: 1 H NMR (300MHz, DMSO-d 6 )δ9.77– 9.47 (m, 2H), 7.71-7.58 (m, 1H), 5.30-5.19 (m, 1H), 3.97-3.58 (m, 4H), 3.52-3.32 (m, 6H), 3.14-2.95 (m, 3H) ), 2.88-2.76 (m, 1H), 1.09 (s, 3H), 0.89 (s, 6H), 0.87 (s, 3H), 0.85 (s, 3H), 0.67 (s, 3H), 0.66 (s, 3H). ESI-MS: m/z 568.5 [M+H] + .
实施例84Example 84
N-(2-(4-甲基-1-哌嗪基)乙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-12)N-(2-(4-Methyl-1-piperazinyl)ethyl)-3β-hydroxy-oleanane-12-ene-28-amide (compound B-12)
Figure PCTCN2020093248-appb-000121
Figure PCTCN2020093248-appb-000121
参照实施例73的方法，将N,N-二甲基乙二胺替换成1-(2-氨乙基)-4-甲基哌嗪，制得化合物B-12： 1H NMR(300MHz,CDCl 3)δ6.55–6.44(m,1H),5.35(s,1H),4.30–4.14(m,1H),3.46–3.31(m,1H),3.28–3.05(m,2H),2.61–2.36(m,10H),2.29(s,3H),1.14(s,3H),0.96(s,3H),0.88(s,9H),0.75(s,3H),0.73(s,3H).ESI-MS:m/z 582.5[M+H] +。 Referring to the method in Example 73, N,N-dimethylethylenediamine was replaced with 1-(2-aminoethyl)-4-methylpiperazine to obtain compound B-12: 1 H NMR (300MHz, CDCl 3 )δ6.55-6.44(m,1H), 5.35(s,1H), 4.30-4.14(m,1H), 3.46-3.31(m,1H), 3.28-3.05(m,2H), 2.61- 2.36(m,10H),2.29(s,3H),1.14(s,3H),0.96(s,3H),0.88(s,9H),0.75(s,3H),0.73(s,3H).ESI -MS: m/z 582.5 [M+H] + .
实施例85Example 85
N-(2-(1,1-二氧代硫代吗啉基)乙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-13)N-(2-(1,1-dioxothiomorpholinyl)ethyl)-3β-hydroxy-oleanane-12-en-28-amide (compound B-13)
Figure PCTCN2020093248-appb-000122
Figure PCTCN2020093248-appb-000122
取齐墩果酸(OA，200mg，0.438mmol)溶于N,N-二甲基甲酰胺(8mL)中，加入三乙胺(243μL，1.752mmol)，室温搅拌下加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(216mg，0.569mmol)，反应1小时后加入化合物4-(2-氨乙基)硫代吗啉-1,1-二氧化物(156mg，0.875mmol)，反应液升温 至60℃。TLC检测反应完全后，用乙酸乙酯(10mL)和水(5mL)处理反应液，水层用乙酸乙酯(10mL×2)萃取，合并有机层，水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝50：1)纯化，得化合物B-13(白色固体，175mg，产率65％)： 1H NMR(300MHz,CDCl 3)δ6.22(s,1H),5.35(s,1H),3.49(dd,J＝13.6,6.2Hz,1H),3.20(d,J＝4.8Hz,2H),3.05(s,8H),2.63(s,2H),2.49(d,J＝11.0Hz,1H),1.17(s,3H),0.99(s,2H),0.90(s,6H),0.78(s,2H),0.76(s,3H).ESI-MS:m/z 617.5[M+H] +。 Dissolve oleanolic acid (OA, 200mg, 0.438mmol) in N,N-dimethylformamide (8mL), add triethylamine (243μL, 1.752mmol), add 2-(7-oxidation) with stirring at room temperature Benzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (216mg, 0.569mmol), after 1 hour of reaction, add compound 4-(2-aminoethyl)thio The temperature of the reaction solution was raised to 60°C with phytoline-1,1-dioxide (156mg, 0.875mmol). After the completion of the reaction was detected by TLC, the reaction solution was treated with ethyl acetate (10 mL) and water (5 mL), the aqueous layer was extracted with ethyl acetate (10 mL×2), the organic layers were combined, washed with water (5 mL×2), and anhydrous sulfuric acid Dry sodium, filter, concentrate the filtrate, and purify by silica gel column chromatography (dichloromethane: methanol = 50:1) to obtain compound B-13 (white solid, 175 mg, yield 65%): 1 H NMR (300MHz, CDCl 3 )δ6.22(s,1H),5.35(s,1H), 3.49(dd,J=13.6,6.2Hz,1H), 3.20(d,J=4.8Hz,2H),3.05(s,8H) ,2.63(s,2H),2.49(d,J=11.0Hz,1H),1.17(s,3H),0.99(s,2H),0.90(s,6H),0.78(s,2H),0.76( s, 3H). ESI-MS: m/z 617.5 [M+H] + .
实施例86Example 86
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-(2S)-羧基-四氢吡咯(化合物B-16)N-(3β-hydroxy-oleanorane-12-ene-28-acyl)-(2S)-carboxy-tetrahydropyrrole (compound B-16)
Figure PCTCN2020093248-appb-000123
Figure PCTCN2020093248-appb-000123
参照实施例73的方法，将N,N-二甲基乙二胺替换成L-脯氨酸苄酯盐酸盐，制得化合物XII-1。Referring to the method of Example 73, N,N-dimethylethylenediamine was replaced with L-proline benzyl ester hydrochloride to prepare compound XII-1.
取化合物XII-1(120mg，0.186mmol)溶于四氢呋喃(1mL)和甲醇(1mL)的混合溶液中，加入2N氢氧化钠溶液(1mL)，80℃下反应2小时。TLC检测反应完全后，减压蒸除溶剂，用1N盐酸溶液调节pH至弱酸性，二氯甲烷萃取(5mL×2)，水洗(5mL×2)有机相，饱和食盐水洗涤(5mL×2)，无水硫酸钠干燥，减压蒸除溶剂，残余物用二氯甲烷和乙腈的混合溶剂(5.5mL，v：v＝10：1)于超声中处理，析出白色固体，抽滤，滤饼用二氯甲烷和乙腈的混合溶剂(5.5mL，v：v＝10：1)洗涤两次，红外下干燥，得化合物B-16(白色固体，88mg，产率85％)： 1H NMR(300MHz,DMSO-d 6)δ5.32–5.24(m,1H),4.66–4.53(m,1H),3.85–3.67(m,1H),3.53(q,J＝9.3Hz,1H),3.26–3.06(m,2H),2.40(dd,J＝13.0,2.5Hz,1H),1.15(s,3H),0.98(s,3H),0.94(s,3H),0.91(s,3H),0.90(s,3H),0.77(s,3H),0.71(s,3H).ESI-MS:m/z 552.3[M-H] -。 Take compound XII-1 (120 mg, 0.186 mmol) and dissolve it in a mixed solution of tetrahydrofuran (1 mL) and methanol (1 mL), add 2N sodium hydroxide solution (1 mL), and react at 80° C. for 2 hours. After the completion of the reaction detected by TLC, the solvent was evaporated under reduced pressure, the pH was adjusted to weak acidity with 1N hydrochloric acid solution, extracted with dichloromethane (5mL×2), washed with water (5mL×2) the organic phase, washed with saturated brine (5mL×2) , Dried over anhydrous sodium sulfate, distilled off the solvent under reduced pressure, the residue was treated with a mixed solvent of dichloromethane and acetonitrile (5.5mL, v:v=10:1) under ultrasound, a white solid precipitated, filtered with suction, and filter cake Wash twice with a mixed solvent of dichloromethane and acetonitrile (5.5mL, v:v=10:1), and dry under infrared to obtain compound B-16 (white solid, 88mg, yield 85%): 1 H NMR( 300MHz, DMSO-d 6 )δ5.32–5.24(m,1H), 4.66–4.53(m,1H), 3.85–3.67(m,1H), 3.53(q,J=9.3Hz,1H), 3.26– 3.06 (m, 2H), 2.40 (dd, J = 13.0, 2.5 Hz, 1H), 1.15 (s, 3H), 0.98 (s, 3H), 0.94 (s, 3H), 0.91 (s, 3H), 0.90 (s, 3H), 0.77 (s, 3H), 0.71 (s, 3H). ESI-MS: m/z 552.3 [MH] - .
实施例87Example 87
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-甲基-哌嗪(化合物B-17)N-(3β-hydroxy-oleanane-12-ene-28-acyl)-4-methyl-piperazine (compound B-17)
Figure PCTCN2020093248-appb-000124
Figure PCTCN2020093248-appb-000124
参照实施例73的方法，将N,N-二甲基乙二胺替换成N-甲基哌嗪，制得化合物B-17： 1H NMR(300MHz,CDCl 3)δ5.30–5.24(m,1H),3.76–3.58(m,4H),3.26–3.18(m,1H),3.10(d,J＝12.3Hz,1H),2.45–2.33(m,4H),2.31(s,3H),1.15(s,3H),1.00(s,3H),0.95(s,3H),0.92(s,3H),0.91(s,3H),0.80(s,3H),0.76(s,3H).ESI-MS:m/z 539.5[M+H] +。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with N-methylpiperazine to obtain compound B-17: 1 H NMR (300MHz, CDCl 3 )δ5.30–5.24(m ,1H), 3.76–3.58(m,4H), 3.26–3.18(m,1H), 3.10(d,J=12.3Hz,1H), 2.45–2.33(m,4H), 2.31(s,3H), 1.15(s,3H),1.00(s,3H),0.95(s,3H),0.92(s,3H),0.91(s,3H),0.80(s,3H),0.76(s,3H).ESI -MS: m/z 539.5 [M+H] + .
实施例88Example 88
N-(2-(4-甲基-1-哌嗪基)丙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-18)N-(2-(4-Methyl-1-piperazinyl)propyl)-3β-hydroxy-oleanane-12-ene-28-amide (compound B-18)
Figure PCTCN2020093248-appb-000125
Figure PCTCN2020093248-appb-000125
参照实施例85的方法，将4-(2-氨乙基)硫代吗啉-1,1-二氧化物替换成1-(3-氨丙基)-4-甲基哌嗪，制得化合物B-19： 1H NMR(300MHz,CDCl 3)δ6.41–6.18(m,1H),5.41–5.27(m,1H),3.54–3.35(m,1H),3.29–3.15(m,1H),3.15–2.99(m,1H),2.70–2.33(m,11H),2.29(s,3H),1.16(s,3H),0.99(s,3H),0.91(s,9H),0.79(s,3H),0.76(s,3H).ESI-MS:m/z 596.6[M+H] +。 According to the method of Example 85, 4-(2-aminoethyl)thiomorpholine-1,1-dioxide was replaced with 1-(3-aminopropyl)-4-methylpiperazine to prepare Compound B-19: 1 H NMR (300MHz, CDCl 3 ) δ6.41–6.18(m,1H), 5.41–5.27(m,1H), 3.54–3.35(m,1H), 3.29–3.15(m,1H) ), 3.15–2.99(m, 1H), 2.70–2.33(m, 11H), 2.29(s, 3H), 1.16(s, 3H), 0.99(s, 3H), 0.91(s, 9H), 0.79( s, 3H), 0.76 (s, 3H). ESI-MS: m/z 596.6 [M+H] + .
实施例89Example 89
N-(2-(1-四氢吡咯基)丙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-19)N-(2-(1-Tetrahydropyrrolyl)propyl)-3β-hydroxy-oleanorane-12-en-28-amide (compound B-19)
Figure PCTCN2020093248-appb-000126
Figure PCTCN2020093248-appb-000126
参照实施例85的方法，将4-(2-氨乙基)硫代吗啉-1,1-二氧化物替换成1-(3-氨基丙基)吡咯烷，制得化合物B-19： 1H NMR(300MHz,CDCl 3)δ5.63–5.33(m,1H),3.65–3.33(m,4H),3.30–2.98(m,6H),2.75–2.43(m,1H),0.99(s,3H),0.94(s,3H),0.91(s,6H),0.78(s,3H),0.71(s,3H).ESI-MS:m/z 567.5[M+H] +。 Referring to the method of Example 85, 4-(2-aminoethyl)thiomorpholine-1,1-dioxide was replaced with 1-(3-aminopropyl)pyrrolidine to obtain compound B-19: 1 H NMR (300MHz, CDCl 3 ) δ5.63-5.33(m,1H), 3.65-3.33(m,4H), 3.30-2.98(m,6H), 2.75-2.43(m,1H), 0.99(s ,3H),0.94(s,3H),0.91(s,6H),0.78(s,3H),0.71(s,3H). ESI-MS: m/z 567.5[M+H] + .
实施例90Example 90
N-(2-(1,1-二氧代硫代吗啉基)丙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-20)N-(2-(1,1-dioxothiomorpholinyl)propyl)-3β-hydroxy-oleanane-12-en-28-amide (compound B-20)
Figure PCTCN2020093248-appb-000127
Figure PCTCN2020093248-appb-000127
参照实施例85的方法，将4-(2-氨乙基)硫代吗啉-1,1-二氧化物替换成4-(3-氨丙基)硫代吗啉-1,1-二氧化物制得化合物B-20： 1H NMR(300MHz,CDCl 3)δ6.10–5.98(m,1H),5.43–5.28(m,1H),3.50–3.33(m,1H),3.30–3.18(m,1H), 3.12–2.90(m,9H),2.59–2.42(m,3H),1.17(s,3H),0.99(s,3H),0.91(s,9H),0.79(s,3H),0.76(s,3H).ESI-MS:m/z 631.5[M+H] +。 Referring to the method of Example 85, 4-(2-aminoethyl)thiomorpholine-1,1-dioxide was replaced with 4-(3-aminopropyl)thiomorpholine-1,1-dioxide Compound B-20 was prepared by oxide: 1 H NMR (300MHz, CDCl 3 ) δ 6.10–5.98 (m, 1H), 5.43–5.28 (m, 1H), 3.50–3.33 (m, 1H), 3.30–3.18 (m,1H), 3.12-2.90(m,9H), 2.59-2.42(m,3H), 1.17(s,3H), 0.99(s,3H), 0.91(s,9H), 0.79(s,3H) ), 0.76(s, 3H). ESI-MS: m/z 631.5[M+H] + .
实施例91Example 91
N-(2-(1-哌啶基)丙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-22)N-(2-(1-piperidinyl)propyl)-3β-hydroxy-oleanorane-12-en-28-amide (compound B-22)
Figure PCTCN2020093248-appb-000128
Figure PCTCN2020093248-appb-000128
参照实施例85的方法，将4-(2-氨乙基)硫代吗啉-1,1-二氧化物替换成1-(3-氨基丙基)哌啶，制得化合物B-22： 1H NMR(300MHz,DMSO-d 6)δ8.82(s,1H),7.39(s,1H),5.22(s,1H),4.22(s,1H),3.43–3.32(m,2H),3.12–3.02(2H),3.01–2.95(s,2H),2.89–2.79(m,2H),1.09(s,3H),0.89(s,6H),0.88(s,3H),0.85(s,3H),0.68(s,3H),0.67(s,3H).ESI-MS:m/z 581.5[M+H] +。 Referring to the method of Example 85, 4-(2-aminoethyl)thiomorpholine-1,1-dioxide was replaced with 1-(3-aminopropyl)piperidine to prepare compound B-22: 1 H NMR(300MHz,DMSO-d 6 )δ8.82(s,1H),7.39(s,1H),5.22(s,1H),4.22(s,1H),3.43-3.32(m,2H), 3.12–3.02(2H), 3.01–2.95(s, 2H), 2.89–2.79(m, 2H), 1.09(s, 3H), 0.89(s, 6H), 0.88(s, 3H), 0.85(s, 3H), 0.68 (s, 3H), 0.67 (s, 3H). ESI-MS: m/z 581.5 [M+H] + .
实施例92Example 92
N-(2-(三甲基铵基)乙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺溴化物(化合物B-23)N-(2-(Trimethylammonium)ethyl)-3β-hydroxy-oleanorane-12-ene-28-amide bromide (Compound B-23)
Figure PCTCN2020093248-appb-000129
Figure PCTCN2020093248-appb-000129
取齐墩果酸(OA，100mg，0.219mmol)溶于N,N-二甲基甲酰胺(8mL)中，加入三乙胺(122μL，0.876mmol)，室温搅拌下加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU,108mg，0.284mmol)，反应1小时后加入化合物I-4(116mg，0.438mmol)，反应液升温至50℃。TLC检测反应完全后，用二氯甲烷(10mL)和去离子水(5mL)处理反应液，水层用二氯甲烷(10mL×2)萃取，合并有机层，去离子水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝10：1)纯化，得化合物B-23(白色固体，43mg，产率32％)： 1H NMR(300MHz,methanol-d 4)δ7.75–7.66(m,1H),5.46–5.39(m,1H),3.79–3.54(m,2H),3.47(t,J＝6.8Hz,2H),3.30–3.14(m,10H),2.91–2.80(m,1H),1.23(s,3H),1.02(s,3H),1.00(s,6H),0.97(s,3H),0.83(s,3H),0.82(s,3H).ESI-MS:m/z 541.5[M-Br] +。 Dissolve oleanolic acid (OA, 100mg, 0.219mmol) in N,N-dimethylformamide (8mL), add triethylamine (122μL, 0.876mmol), add 2-(7-oxidation) with stirring at room temperature Benzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU, 108mg, 0.284mmol), after 1 hour of reaction, compound I-4 (116mg, 0.438mmol) was added, The reaction solution was heated to 50°C. After the completion of the reaction detected by TLC, the reaction solution was treated with dichloromethane (10mL) and deionized water (5mL), the aqueous layer was extracted with dichloromethane (10mL×2), the organic layers were combined and washed with deionized water (5mL×2) , Dried over anhydrous sodium sulfate, filtered, concentrated the filtrate, and purified by silica gel column chromatography (dichloromethane: methanol = 10:1) to obtain compound B-23 (white solid, 43mg, yield 32%): 1 H NMR (300MHz, methanol-d 4 )δ7.75–7.66(m,1H), 5.46–5.39(m,1H), 3.79–3.54(m,2H), 3.47(t,J=6.8Hz,2H), 3.30 --3.14(m,10H),2.91--2.80(m,1H),1.23(s,3H),1.02(s,3H),1.00(s,6H),0.97(s,3H),0.83(s,3H) ), 0.82(s, 3H). ESI-MS: m/z 541.5[M-Br] + .
实施例93Example 93
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-哌嗪(化合物B-25)N-(3β-Hydroxy-oleanane-12-ene-28-acyl)-piperazine (Compound B-25)
Figure PCTCN2020093248-appb-000130
Figure PCTCN2020093248-appb-000130
参照实施例73的方法，将N,N-二甲基乙二胺替换成1-苄基哌嗪，制得化合物XIII-1。According to the method of Example 73, N,N-dimethylethylenediamine was replaced with 1-benzylpiperazine to prepare compound XIII-1.
取化合物XIII-1(100mg，0.163mmol)溶于四氢呋喃(8mL)中，加入10％钯碳(10mg)，氢气氛围下室温搅拌反应过夜。TLC检测反应完全后，过滤硅藻土，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物B-25(白色固体，61mg，产率72％)： 1H NMR(300MHz,DMSO-d 6)δ9.28(s,2H),5.15–5.02(m,1H),4.34–4.24(m,1H),3.86–3.60(m,4H),3.09–2.87(m,6H),2.12–1.95(m,1H),1.09(s,3H),0.88(s,9H),0.84(s,3H),0.67(s,3H),0.64(s,3H).ESI-MS:m/z 525.5[M+H] +。 The compound XIII-1 (100 mg, 0.163 mmol) was dissolved in tetrahydrofuran (8 mL), 10% palladium on carbon (10 mg) was added, and the reaction was stirred overnight at room temperature under a hydrogen atmosphere. After the completion of the reaction detected by TLC, Celite was filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound B-25 (white solid, 61 mg, yield 72%): 1 H NMR(300MHz, DMSO-d 6 )δ9.28(s,2H), 5.15-5.02(m,1H), 4.34-4.24(m,1H), 3.86-3.60(m,4H), 3.09-2.87( m,6H),2.12-1.95(m,1H),1.09(s,3H),0.88(s,9H),0.84(s,3H),0.67(s,3H),0.64(s,3H).ESI -MS: m/z 525.5 [M+H] + .
实施例94Example 94
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-四氢吡咯(化合物B-26)N-(3β-hydroxy-oleanorane-12-ene-28-acyl)-tetrahydropyrrole (compound B-26)
Figure PCTCN2020093248-appb-000131
Figure PCTCN2020093248-appb-000131
参照实施例73的方法，将N,N-二甲基乙二胺替换成四氢吡咯，制得化合物B-26： 1H NMR(300MHz,CDCl 3)δ7.10–6.85(m,1H),5.44–5.22(m,1H),3.63–3.42(m,1H),3.29–3.13(m,1H),3.12–2.93(m,1H),2.67–2.48(m,2H),2.48–2.34(m,3H),2.27(s,6H),1.15(s,4H),0.99(s,4H),0.91(s,9H),0.78(s,3H),0.76(s,3H).ESI-MS:m/z 541.5[M+H] +。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with tetrahydropyrrole to obtain compound B-26: 1 H NMR (300MHz, CDCl 3 )δ7.10-6.85(m,1H) ,5.44-5.22(m,1H),3.63-3.42(m,1H), 3.29-3.13(m,1H), 3.12-2.93(m,1H), 2.67-2.48(m,2H), 2.48-2.34( m,3H),2.27(s,6H),1.15(s,4H),0.99(s,4H),0.91(s,9H),0.78(s,3H),0.76(s,3H).ESI-MS :m/z 541.5[M+H] + .
实施例95Example 95
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-(3S)-氨基-四氢吡咯(化合物B-27)N-(3β-Hydroxy-oleanorane-12-ene-28-acyl)-(3S)-amino-tetrahydropyrrole (Compound B-27)
Figure PCTCN2020093248-appb-000132
Figure PCTCN2020093248-appb-000132
Figure PCTCN2020093248-appb-000133
Figure PCTCN2020093248-appb-000133
参照实施例73的方法，将N,N-二甲基乙二胺替换成(S)-3-N-苄氧羰基氨基吡咯烷制得化合物XIV-1。According to the method of Example 73, N,N-dimethylethylenediamine was replaced with (S)-3-N-benzyloxycarbonylaminopyrrolidine to prepare compound XIV-1.
取化合物XIV-1(216mg，0.328mmol)溶于四氢呋喃(8mL)中，加入10％钯碳(20mg)，氢气氛围下室温搅拌反应过夜。TLC检测反应完全后，过滤硅藻土，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物B-27(白色固体，143mg，产率83％)： 1H NMR(300MHz,CDCl 3)δ5.29–5.22(m,1H),3.74–3.61(m,2H),3.61–3.49(m,2H),3.37–3.25(m,1H),3.25–3.07(m,2H),1.14(s,3H),0.98(s,3H),0.94(s,3H),0.89(s,6H),0.77(s,3H),0.72(s,3H).ESI-MS:m/z 525.5[M+H] +。 The compound XIV-1 (216 mg, 0.328 mmol) was dissolved in tetrahydrofuran (8 mL), 10% palladium on carbon (20 mg) was added, and the reaction was stirred overnight at room temperature under a hydrogen atmosphere. After the completion of the reaction detected by TLC, Celite was filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound B-27 (white solid, 143 mg, yield 83%): 1 H NMR (300MHz, CDCl 3 ) δ 5.29-5.22 (m, 1H), 3.74-3.61 (m, 2H), 3.61--3.49 (m, 2H), 3.37-3.25 (m, 1H), 3.25-3.07 ( m,2H),1.14(s,3H),0.98(s,3H),0.94(s,3H),0.89(s,6H),0.77(s,3H),0.72(s,3H).ESI-MS :m/z 525.5[M+H] + .
实施例96Example 96
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-(3R)-氨基-四氢吡咯(化合物B-28)N-(3β-hydroxy-oleanolane-12-ene-28-acyl)-(3R)-amino-tetrahydropyrrole (compound B-28)
Figure PCTCN2020093248-appb-000134
Figure PCTCN2020093248-appb-000134
参照实施例95的方法，将(S)-3-N-苄氧羰基氨基吡咯烷替换成(R)-3-N-苄氧羰基氨基吡咯烷制得化合物B-28： 1H NMR(300MHz,CDCl 3)δ5.29–5.20(m,1H),3.90–3.72(m,1H),3.72–3.59(m,1H),3.58–3.40(m,2H),3.26–3.04(m,3H),2.13–1.99(m,2H),1.13(s,3H),0.98(s,3H),0.93(s,3H),0.89(s,6H),0.77(s,3H),0.71(s,3H).ESI-MS:m/z 525.5[M+H] +。 Referring to the method of Example 95, (S)-3-N-benzyloxycarbonylaminopyrrolidine was replaced with (R)-3-N-benzyloxycarbonylaminopyrrolidine to prepare compound B-28: 1 H NMR (300MHz ,CDCl 3 )δ5.29--5.20(m,1H),3.90-3.72(m,1H),3.72-3.59(m,1H),3.58-3.40(m,2H), 3.26-3.04(m,3H) ,2.13-1.99(m,2H),1.13(s,3H),0.98(s,3H),0.93(s,3H),0.89(s,6H),0.77(s,3H),0.71(s,3H) ). ESI-MS: m/z 525.5 [M+H] + .
实施例97Example 97
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-(3S)-羟基-四氢吡咯(化合物B-29)N-(3β-hydroxy-oleanorane-12-ene-28-acyl)-(3S)-hydroxy-tetrahydropyrrole (compound B-29)
Figure PCTCN2020093248-appb-000135
Figure PCTCN2020093248-appb-000135
参照实施例73的方法，将N,N-二甲基乙二胺替换成(S)-3-吡咯烷醇，制得化合物B-29： 1H NMR(300MHz,CDCl 3)δ5.26(m,1H),4.50–4.36(m,1H),3.75–3.61(m,2H),3.61–3.49(m,2H),3.24–3.09(m,2H),1.13(s,3H),0.97(s,3H),0.93(s,3H),0.89(s,3H),0.88(s,3H),0.76(s,3H),0.72(s,3H).ESI-MS:m/z 548.4[M+Na] +。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with (S)-3-pyrrolidinol to prepare compound B-29: 1 H NMR (300MHz, CDCl 3 )δ 5.26( m, 1H), 4.50--4.36 (m, 1H), 3.75--3.61 (m, 2H), 3.61--3.49 (m, 2H), 3.24--3.09 (m, 2H), 1.13 (s, 3H), 0.97 ( s,3H),0.93(s,3H),0.89(s,3H),0.88(s,3H),0.76(s,3H),0.72(s,3H).ESI-MS:m/z 548.4(M +Na] + .
实施例98Example 98
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-(3R)-羟基-四氢吡咯(化合物B-30)N-(3β-Hydroxy-oleanan-12-ene-28-acyl)-(3R)-Hydroxy-tetrahydropyrrole (Compound B-30)
Figure PCTCN2020093248-appb-000136
Figure PCTCN2020093248-appb-000136
参照实施例73的方法，将N,N-二甲基乙二胺替换成(R)-3-吡咯烷醇，制得化合物B-30： 1H NMR(300MHz,CDCl 3)δ5.24(m,1H),4.49–4.31(m,1H),3.81–3.57(m,3H),3.56–3.43(m,1H),3.29–3.13(m,1H),3.14–2.95(m,1H),2.56–2.16(m,1H),1.13(s,3H),0.97(s,3H),0.93(s,3H),0.89(s,3H),0.88(s,3H),0.76(s,3H),0.70(s,3H).ESI-MS:m/z 548.4[M+Na] +。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with (R)-3-pyrrolidinol to obtain compound B-30: 1 H NMR (300MHz, CDCl 3 )δ5.24( m,1H), 4.49–4.31(m,1H), 3.81–3.57(m,3H), 3.56–3.43(m,1H), 3.29–3.13(m,1H), 3.14–2.95(m,1H), 2.56--2.16(m,1H),1.13(s,3H),0.97(s,3H),0.93(s,3H),0.89(s,3H),0.88(s,3H),0.76(s,3H) ,0.70(s,3H). ESI-MS: m/z 548.4[M+Na] + .
实施例99Example 99
3β-羟基-齐墩果烷-12-烯-28-酸2-(1-哌啶基)乙酯(化合物B-31)3β-Hydroxy-oleanorane-12-en-28-acid 2-(1-piperidinyl) ethyl ester (Compound B-31)
Figure PCTCN2020093248-appb-000137
Figure PCTCN2020093248-appb-000137
取齐墩果酸(OA，5g，10.9mmol)溶于N,N-二甲基甲酰胺(40mL)中，依次加入1,2-二溴乙烷(10.2mL，109mmol)、碳酸钾(1.506g，10.9mmol)和乙腈(4mL)，将反应液升温至50℃，搅拌反应3小时。TLC检测反应完全后，用二氯甲烷(100mL)和水(100mL)处理反应液，水层用二氯甲烷(50mL)萃取，合并有机相，水(100mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝10：1)纯化，得化合物XV-1(白色固体，4.44g，产率72％)。Dissolve oleanolic acid (OA, 5g, 10.9mmol) in N,N-dimethylformamide (40mL), add 1,2-dibromoethane (10.2mL, 109mmol), potassium carbonate (1.506 g, 10.9 mmol) and acetonitrile (4 mL), the reaction solution was heated to 50° C., and the reaction was stirred for 3 hours. After the completion of the reaction detected by TLC, the reaction solution was treated with dichloromethane (100mL) and water (100mL), the aqueous layer was extracted with dichloromethane (50mL), the organic phases were combined, washed with water (100mL×2), and dried over anhydrous sodium sulfate , Filtration, concentration of the filtrate, and purification by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound XV-1 (white solid, 4.44 g, yield 72%).
取化合物XV-1(200mg，0.35mmol)溶于无水N,N-二甲基甲酰胺(10mL)中，依次加入哌啶(70μL，0.70mmol)和碳酸钾(48mg，0.35mmol)，将反应液升温至50℃，搅拌反应。TLC检测反应完全后，用二氯甲烷(10mL)和水(5mL)处理反应液，水层用二氯甲烷(10mL×2)萃取，合并有机相，水(10mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝30：1)纯化，得化合物B-31(白色固体，47mg，产率24％)： 1H NMR(300MHz,CDCl 3)δ5.29–5.23(m,1H),4.27–4.09(m,2H),3.26–3.15(m,1H),2.89– 2.79(m,1H),2.63(t,J＝5.5Hz,2H),2.54–2.42(m,4H),1.13(s,3H),0.98(s,3H),0.92(s,3H),0.90(s,6H),0.78(s,3H),0.73(s,3H).ESI-MS:m/z 568.6[M+H] +。 Dissolve compound XV-1 (200mg, 0.35mmol) in anhydrous N,N-dimethylformamide (10mL), add piperidine (70μL, 0.70mmol) and potassium carbonate (48mg, 0.35mmol) in sequence, The reaction solution was heated to 50°C, and the reaction was stirred. After the completion of the reaction was detected by TLC, the reaction solution was treated with dichloromethane (10mL) and water (5mL), the aqueous layer was extracted with dichloromethane (10mL×2), the organic phases were combined, washed with water (10mL×2), and anhydrous sulfuric acid The sodium was dried, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain compound B-31 (white solid, 47 mg, yield 24%): 1 H NMR (300MHz, CDCl 3 )δ5.29–5.23(m,1H), 4.27–4.09(m,2H), 3.26–3.15(m,1H), 2.89– 2.79(m,1H), 2.63(t,J=5.5Hz,2H ),2.54--2.42(m,4H),1.13(s,3H),0.98(s,3H),0.92(s,3H),0.90(s,6H),0.78(s,3H),0.73(s, 3H). ESI-MS: m/z 568.6 [M+H] + .
实施例100Example 100
3β-羟基-齐墩果烷-12-烯-28-酸2-(4-吗啉基)乙酯(化合物B-32)3β-Hydroxy-oleanolane-12-ene-28-acid 2-(4-morpholinyl) ethyl ester (Compound B-32)
Figure PCTCN2020093248-appb-000138
Figure PCTCN2020093248-appb-000138
参照实施例99的方法,将哌啶替换成吗啉，制得化合物B-32： 1H NMR(300MHz,CDCl 3)δ5.30–5.18(m,1H),4.70–4.52(m,2H),4.42–4.11(m,2H),4.11–3.79(m,2H),3.63–3.31(m,2H),3.31–3.08(m,3H),3.04–2.84(m,2H),2.78(d,J＝12.5Hz,1H),1.14(s,3H),0.99(s,3H),0.91(s,9H),0.78(s,3H),0.70(s,3H).ESI-MS:m/z 570.6[M+H] +。 Referring to the method of Example 99, the piperidine was replaced with morpholine to obtain compound B-32: 1 H NMR (300MHz, CDCl 3 ) δ5.30–5.18(m,1H), 4.70–4.52(m,2H) ,4.42–4.11(m,2H),4.11–3.79(m,2H),3.63–3.31(m,2H),3.31–3.08(m,3H),3.04–2.84(m,2H),2.78(d, J = 12.5Hz, 1H), 1.14 (s, 3H), 0.99 (s, 3H), 0.91 (s, 9H), 0.78 (s, 3H), 0.70 (s, 3H). ESI-MS: m/z 570.6[M+H] + .
实施例101Example 101
3β-羟基-齐墩果烷-12-烯-28-酸2-(1-四氢吡咯基)乙酯(化合物B-33)3β-Hydroxy-oleanolane-12-ene-28-acid 2-(1-tetrahydropyrrolyl) ethyl ester (Compound B-33)
Figure PCTCN2020093248-appb-000139
Figure PCTCN2020093248-appb-000139
参照实施例99的方法，将哌啶替换成四氢吡咯，制得化合物B-33： 1H NMR(300MHz,CDCl 3)δ5.31–5.21(m,1H),4.67–4.48(m,2H),3.90–3.71(m,2H),3.39–3.12(m,3H),2.92–2.72(m,3H),2.35–2.16(m,2H),2.16–2.02(m,2H),2.02–1.92(m,1H),1.91–1.82(m,2H),1.13(s,3H),0.98(s,3H),0.90(s,9H),0.77(s,3H),0.69(s,3H).ESI-MS:m/z 554.6[M+H] +。 According to the method of Example 99, the piperidine was replaced with tetrahydropyrrole to obtain compound B-33: 1 H NMR (300MHz, CDCl 3 ) δ5.31–5.21(m,1H), 4.67–4.48(m,2H ), 3.90–3.71(m,2H), 3.39–3.12(m,3H), 2.92–2.72(m,3H), 2.35–2.16(m,2H), 2.16–2.02(m,2H), 2.02–1.92 (m, 1H), 1.91--1.82 (m, 2H), 1.13 (s, 3H), 0.98 (s, 3H), 0.90 (s, 9H), 0.77 (s, 3H), 0.69 (s, 3H). ESI-MS: m/z 554.6 [M+H] + .
实施例102Example 102
N-(2-(乙酰氨基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-38)N-(2-(Acetylamino)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanolane-12-ene-28-amide (compound B-38)
Figure PCTCN2020093248-appb-000140
Figure PCTCN2020093248-appb-000140
参照实施例85的方法，将4-(2-氨乙基)硫代吗啉-1,1-二氧化物替换成N-乙酰基乙二胺，制得化合物XVI-1。According to the method of Example 85, 4-(2-aminoethyl)thiomorpholine-1,1-dioxide was replaced with N-acetylethylenediamine to prepare compound XVI-1.
将化合物XVI-1(120mg，0.222mmol)溶于无水吡啶(8mL)中，加入邻苯二甲酸酐(329mg，2.22mmol)和4-二甲氨基吡啶(28mg，0.222mmol)，反应液升温至115℃搅拌过夜。TLC检测反应完全后，加入二氯甲烷(20mL)稀释反应液，反应液依次用1N稀盐酸(10mL×3)和水(10mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物B-38(白色固体，96mg，产率63％)：制得化合物B-38： 1H NMR(300MHz,methanol-d 4)δ7.79–7.72(m,1H),7.68–7.63(m,1H),7.60–7.52(m,2H),7.35–7.24(m,1H),5.52–5.44(m,1H),5.41–5.34(m,1H),4.77–4.61(m,1H),3.20–3.05(m,1H),2.83–2.65(m,1H),1.21–1.19(m,6H),0.99(s,3H),0.97(s,3H),0.95(s,3H),0.91(s,6H),0.78(s,3H).ESI-MS:m/z 687.5[M-H] -。 Compound XVI-1 (120mg, 0.222mmol) was dissolved in anhydrous pyridine (8mL), phthalic anhydride (329mg, 2.22mmol) and 4-dimethylaminopyridine (28mg, 0.222mmol) were added, and the reaction solution was heated Stir at 115°C overnight. After the completion of the reaction detected by TLC, dichloromethane (20mL) was added to dilute the reaction solution. The reaction solution was washed with 1N dilute hydrochloric acid (10mL×3) and water (10mL×3) successively, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound B-38 (white solid, 96 mg, yield 63%): to obtain compound B-38: 1 H NMR (300MHz, methanol-d) 4 )δ7.79–7.72(m,1H), 7.68–7.63(m,1H), 7.60–7.52(m,2H), 7.35–7.24(m,1H), 5.52–5.44(m,1H), 5.41 --5.34(m,1H),4.77–4.61(m,1H), 3.20–3.05(m,1H), 2.83–2.65(m,1H), 1.21–1.19(m,6H),0.99(s,3H) , 0.97 (s, 3H), 0.95 (s, 3H), 0.91 (s, 6H), 0.78 (s, 3H). ESI-MS: m/z 687.5 [MH] - .
实施例103Example 103
N-(2-(1-哌啶基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-39)N-(2-(1-piperidinyl)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-12-en-28-amide (compound B-39)
Figure PCTCN2020093248-appb-000141
Figure PCTCN2020093248-appb-000141
参照实施例102的方法，将N-乙酰基乙二胺替换成1-(2-氨乙基)哌啶，制得化合物B-39： 1H NMR(300MHz,CDCl 3)δ7.73–7.61(m,2H),7.50–7.40(m,1H),7.39–7.30(m,1H),7.09(d,J＝13.4Hz,1H),5.41–5.25(m,1H),4.79–4.55(m,1H),3.88–3.55(m,3H),3.17–2.76(m,6H),1.17(s,3H),0.99(s,3H),0.95(s,6H),0.89(s,3H),0.87(s,3H),0.77(s,3H).ESI-MS:m/z 715.7[M+H] +。 Referring to the method of Example 102, N-acetylethylenediamine was replaced with 1-(2-aminoethyl)piperidine to obtain compound B-39: 1 H NMR (300MHz, CDCl 3 ) δ7.73-7.61 (m,2H),7.50–7.40(m,1H),7.39–7.30(m,1H), 7.09(d,J=13.4Hz,1H), 5.41–5.25(m,1H), 4.79–4.55(m ,1H),3.88-3.55(m,3H),3.17-2.76(m,6H),1.17(s,3H),0.99(s,3H),0.95(s,6H),0.89(s,3H), 0.87 (s, 3H), 0.77 (s, 3H). ESI-MS: m/z 715.7 [M+H] + .
实施例104Example 104
N-(2-(4-吗啉基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-40)N-(2-(4-morpholinyl)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanolic-12-en-28-amide (compound B-40)
Figure PCTCN2020093248-appb-000142
Figure PCTCN2020093248-appb-000142
参照实施例102的方法，将N-乙酰基乙二胺替换成N-(2-氨基乙基)吗啉，制得化合物B-40：1H NMR(300MHz,methanol-d 4)δ7.74–7.63(m,2H),7.60–7.46(m,2H),5.45–5.32(m,1H),4.74–4.62(m,1H),3.82–3.72(m,4H),3.51–3.38(m,1H),3.38–3.24(m,3H),2.83–2.58(m,7H),1.21(s,3H),1.01(s,3H),0.98(s,3H),0.94(s,6H),0.92(s,3H),0.81(s,3H).ESI-MS:m/z 718.6[M+H] +。 Referring to the method of Example 102, N-acetylethylenediamine was replaced with N-(2-aminoethyl)morpholine to obtain compound B-40: 1H NMR (300MHz, methanol-d 4 )δ7.74– 7.63 (m, 2H), 7.60 - 7.46 (m, 2H), 5.45 - 5.32 (m, 1H), 4.74 - 4.62 (m, 1H), 3.82 - 3.72 (m, 4H), 3.51 - 3.38 (m, 1H) ), 3.38–3.24(m,3H), 2.83–2.58(m,7H), 1.21(s, 3H), 1.01(s, 3H), 0.98(s, 3H), 0.94(s, 6H), 0.92( s, 3H), 0.81 (s, 3H). ESI-MS: m/z 718.6 [M+H] + .
实施例105Example 105
N-(2-(1-四氢吡咯基)乙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-41)N-(2-(1-Tetrahydropyrrolyl)ethyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-12-en-28-amide (compound B-41)
Figure PCTCN2020093248-appb-000143
Figure PCTCN2020093248-appb-000143
参照实施例102的方法，将N-乙酰基乙二胺替换成1-(2-氨乙基)吡咯烷，制得化合物B-41： 1H NMR(300MHz,methanol-d 4)δ7.82–7.61(m,1H),7.59–7.45(m,2H),7.44–7.32(m,1H),5.48(s,1H),5.33(s,1H),4.72–4.64(m,1H),3.67–3.45(m,2H),3.23–3.10(m,2H),2.94–2.76(m,1H),1.20(s,3H),1.00(s,9H),0.93(s,3H),0.90(s,3H),0.80(s,3H).ESI-MS:m/z 702.6[M+H] +。 Referring to the method in Example 102, N-acetylethylenediamine was replaced with 1-(2-aminoethyl)pyrrolidine to obtain compound B-41: 1 H NMR (300MHz, methanol-d 4 )δ7.82 --7.61 (m, 1H), 7.59 - 7.45 (m, 2H), 7.44 - 7.32 (m, 1H), 5.48 (s, 1H), 5.33 (s, 1H), 4.72 - 4.64 (m, 1H), 3.67 --3.45(m,2H),3.23--3.10(m,2H),2.94-2.76(m,1H),1.20(s,3H),1.00(s,9H),0.93(s,3H),0.90(s , 3H), 0.80 (s, 3H). ESI-MS: m/z 702.6 [M+H] + .
实施例106Example 106
N-(2-(4-吗啉基)乙基)-3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-47)N-(2-(4-morpholinyl)ethyl)-3β-(2-(2-acetylaminoethoxyyl)benzoyl)oxy-oleanorane-12-ene-28-amide ( Compound B-47)
Figure PCTCN2020093248-appb-000144
Figure PCTCN2020093248-appb-000144
取齐墩果酸(OA，20g,0.044mol)溶于N,N-二甲基甲酰胺(200mL)中，加入碳酸钾(6g,0.044mol)，搅拌下缓慢滴加溴化苄(8mL，0.044mol)，滴加完毕后将反应液升温至85℃，反应10小时。TLC检测反应完全后，趁热抽滤，滤饼用热的N,N-二甲基甲酰胺(10mL×3)洗涤，滤液冷却至室温后滴入冰水 (100mL×2)中，析出大量白色固体，抽滤，滤饼用水洗(50mL×3)，正己烷洗(50mL×3)，真空干燥，得化合物III-1(白色固体，22.02g，产率92％)。Dissolve oleanolic acid (OA, 20g, 0.044mol) in N,N-dimethylformamide (200mL), add potassium carbonate (6g, 0.044mol), slowly add benzyl bromide (8mL, 0.044 mol). After the dropping, the reaction solution was heated to 85°C and reacted for 10 hours. After the completion of the reaction detected by TLC, it was filtered while hot, the filter cake was washed with hot N,N-dimethylformamide (10mL×3), the filtrate was cooled to room temperature and then dropped into ice water (100mL×2), a large amount of precipitation The white solid was filtered with suction, the filter cake was washed with water (50 mL×3), washed with n-hexane (50 mL×3), and dried under vacuum to obtain compound III-1 (white solid, 22.02 g, yield 92%).
将化合物III-1(1g，1.829mmol)溶于无水吡啶(10mL)中，加入邻苯二甲酸酐(2.708g，18.29mmol)和4-二甲氨基吡啶(223mg，1.829mmol)，反应液升温至115℃搅拌过夜。TLC检测反应完全后，加入二氯甲烷(30mL)稀释反应液，反应液依次用1N稀盐酸(10mL×3)和水(10mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物XVII-2(白色固体，925mg，产率73％)。Compound III-1 (1g, 1.829mmol) was dissolved in anhydrous pyridine (10mL), phthalic anhydride (2.708g, 18.29mmol) and 4-dimethylaminopyridine (223mg, 1.829mmol) were added to the reaction solution The temperature was raised to 115°C and stirred overnight. After the completion of the reaction was detected by TLC, dichloromethane (30mL) was added to dilute the reaction solution. The reaction solution was washed with 1N diluted hydrochloric acid (10mL×3) and water (10mL×3) successively, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Purification by silica gel column chromatography (dichloromethane:methanol=20:1) gave compound XVII-2 (white solid, 925 mg, yield 73%).
取化合物XVII-2(150mg，0.216mmol)溶于无水二氯甲烷(6mL)中，依次加入N,N'-二环己基碳酰亚胺(134mg，0.647mmol)、4-二甲氨基吡啶(27mg，0.216mmol)和N-乙酰乙醇胺(30μL，0.324mmol)，室温搅拌反应。TLC检测反应完全后，抽滤，滤液用水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝3：1)纯化，得化合物XVII-3(白色固体，159mg，产率95％)。Take compound XVII-2 (150mg, 0.216mmol) and dissolve it in dry dichloromethane (6mL), add N,N'-dicyclohexylcarbimide (134mg, 0.647mmol), 4-dimethylaminopyridine in sequence (27mg, 0.216mmol) and N-acetylethanolamine (30μL, 0.324mmol), the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, it was suction filtered, the filtrate was washed with water (5mL×2), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate=3:1) to obtain the compound XVII-3 (white solid, 159 mg, yield 95%).
取化合物XVII-3(159mg，0.204mmol)溶于四氢呋喃(10mL)中，加入10％钯碳(16mg)，氢气氛围下室温搅拌反应过夜。TLC检测反应完全后，过滤硅藻土，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝1：1)纯化，得化合物XVII-4(白色固体，90mg，产率64％)The compound XVII-3 (159 mg, 0.204 mmol) was dissolved in tetrahydrofuran (10 mL), 10% palladium on carbon (16 mg) was added, and the reaction was stirred overnight at room temperature under a hydrogen atmosphere. After the completion of the reaction detected by TLC, Celite was filtered, the filtrate was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound XVII-4 (white solid, 90 mg, yield 64%)
取化合物XVII-4(100mg，0.145mmol)溶于N,N-二甲基甲酰胺(6mL)中，加入三乙胺(40μL，0.290mmol)，室温搅拌下加入2-(7-氧化苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(72mg，0.188mmol)，反应1小时后加入N-(2-氨基乙基)吗啉(23μL，0.174mmol)，反应液升温至50℃。TLC检测反应完全后，用二氯甲烷(10mL)和水(5mL)处理反应液，水层用二氯甲烷(10mL×2)萃取，合并有机层，水(5mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝30：1)纯化，得化合物B-47(白色固体，80mg，产率69％)： 1H NMR(300MHz,CDCl 3)δ7.78(d,J＝6.1Hz,1H),7.66(d,J＝7.9Hz,1H),7.53–7.42(m,2H),5.34–5.23(m,1H),4.73–4.66(m,1H),4.60–4.49(m,2H),3.52–2.98(m,6H),2.89–2.74(m,1H),1.15(s,3H),0.95(s,3H),0.92(s,9H),0.89(s,3H),0.72(s,3H).ESI-MS:m/z 802.7[M+H] +。 Dissolve compound XVII-4 (100mg, 0.145mmol) in N,N-dimethylformamide (6mL), add triethylamine (40μL, 0.290mmol), add 2-(7-oxybenzoic acid) under stirring at room temperature Triazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (72mg, 0.188mmol), after 1 hour of reaction, add N-(2-aminoethyl)morpholine (23μL, 0.174 mmol), the reaction solution was heated to 50°C. After the completion of the reaction detected by TLC, the reaction solution was treated with dichloromethane (10mL) and water (5mL), the aqueous layer was extracted with dichloromethane (10mL×2), the organic layers were combined, washed with water (5mL×2), and anhydrous sulfuric acid Dry with sodium, filter, concentrate the filtrate, and purify by silica gel column chromatography (dichloromethane: methanol=30:1) to obtain compound B-47 (white solid, 80mg, yield 69%): 1 H NMR (300MHz, CDCl 3 )δ7.78(d,J=6.1Hz,1H), 7.66(d,J=7.9Hz,1H), 7.53–7.42(m,2H), 5.34–5.23(m,1H), 4.73–4.66( m,1H), 4.60--4.49(m,2H), 3.52--2.98(m,6H), 2.89--2.74(m,1H), 1.15(s,3H), 0.95(s,3H), 0.92(s, 9H), 0.89 (s, 3H), 0.72 (s, 3H). ESI-MS: m/z 802.7 [M+H] + .
实施例107Example 107
N-(2-(4-吗啉基)乙基)-3β-(2-(甲氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-48)N-(2-(4-morpholinyl)ethyl)-3β-(2-(methoxyacyl)benzoyl)oxy-oleanorane-12-en-28-amide (compound B-48 )
Figure PCTCN2020093248-appb-000145
Figure PCTCN2020093248-appb-000145
参照实施例106的方法，将N-乙酰乙醇胺替换成甲醇，制得化合物B-48： 1H NMR(300MHz,CDCl 3)δ7.77–7.65(m,2H),7.59–7.48(m,2H),6.55–6.45(m,1H),5.43–5.35(m,1H),4.75(dd,J＝11.4,4.5Hz,1H),3.90(s,3H),3.78–3.68(m,4H),3.48–3.37(m,1H),3.28–3.11(m,1H),2.58–2.39(m,7H),1.18(s,3H),0.97(s,3H),0.96(s,3H),0.92(s,9H),0.78(s,3H).ESI-MS:m/z 753.6[M+Na] +。 According to the method of Example 106, N-acetylethanolamine was replaced with methanol to obtain compound B-48: 1 H NMR (300MHz, CDCl 3 ) δ7.77–7.65 (m, 2H), 7.59–7.48 (m, 2H ), 6.55–6.45(m,1H), 5.43–5.35(m,1H), 4.75(dd,J=11.4,4.5Hz,1H), 3.90(s,3H), 3.78–3.68(m,4H), 3.48–3.37(m,1H), 3.28–3.11(m,1H), 2.58–2.39(m,7H), 1.18(s,3H), 0.97(s,3H), 0.96(s,3H), 0.92( s, 9H), 0.78 (s, 3H). ESI-MS: m/z 753.6 [M+Na] + .
实施例108Example 108
N-(2-(4-吗啉基)乙基)-3β-(2-(异丙基氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-49)N-(2-(4-morpholinyl)ethyl)-3β-(2-(isopropyloxyacyl)benzoyl)oxy-oleanorane-12-en-28-amide (compound B -49)
Figure PCTCN2020093248-appb-000146
Figure PCTCN2020093248-appb-000146
参照实施例106的方法，将N-乙酰乙醇胺替换成异丙醇，制得化合物B-49： 1H NMR(300MHz,CDCl 3)δ7.77–7.65(m,2H),7.57–7.48(m,2H),6.59–6.44(m,1H),5.47–5.36(m,1H),5.34–5.20(m,1H),4.77(dd,J＝11.8,4.6Hz,1H),3.83–3.64(m,4H),3.53–3.39(m,1H),3.29–3.14(m,1H),2.62–2.38(m,7H),1.37(d,J＝6.3Hz,6H),1.20(s,3H),1.00(s,3H),0.98(s,3H),0.94(s,9H),0.80(s,3H).ESI-MS:m/z 759.7[M+H] +。 According to the method of Example 106, N-acetylethanolamine was replaced with isopropanol to obtain compound B-49: 1 H NMR (300MHz, CDCl 3 )δ7.77–7.65(m,2H),7.57–7.48(m ,2H),6.59–6.44(m,1H),5.47–5.36(m,1H),5.34–5.20(m,1H),4.77(dd,J=11.8,4.6Hz,1H),3.83–3.64(m ,4H),3.53–3.39(m,1H), 3.29–3.14(m,1H), 2.62–2.38(m,7H), 1.37(d,J=6.3Hz,6H), 1.20(s,3H), 1.00 (s, 3H), 0.98 (s, 3H), 0.94 (s, 9H), 0.80 (s, 3H). ESI-MS: m/z 759.7 [M+H] + .
实施例109Example 109
N-(2-(二甲基氨基)乙基)-3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-50)N-(2-(dimethylamino)ethyl)-3β-(2-(2-acetylaminoethoxyyl)benzoyl)oxy-oleanorane-12-ene-28-amide (compound B-50)
Figure PCTCN2020093248-appb-000147
Figure PCTCN2020093248-appb-000147
参照实施例106的方法，将N-(2-氨基乙基)吗啉替换成N,N-二甲基乙二胺，制得化合物B-50： 1H NMR(300MHz,CDCl 3)δ7.87–7.81(m,1H),7.72–7.65(m,1H),7.65–7.53(m,2H),6.93–6.85(m,1H),6.62–6.50(m,1H),5.49–5.41(m,1H),4.79–4.67(m,1H),4.46(t,J＝4.5Hz,2H),3.71–3.63(m,2H),3.60–3.47(m,1H),3.47–3.34(m,1H),3.04–2.92(m,2H),2.75–2.57(m,7H),2.04(s,3H),1.21 (s,3H),1.00(s,9H),0.96(s,3H),0.94(s,3H),0.81(s,3H).ESI-MS:m/z 760.6[M+H] +。 Referring to the method of Example 106, N-(2-aminoethyl)morpholine was replaced with N,N-dimethylethylenediamine to obtain compound B-50: 1 H NMR (300MHz, CDCl 3 )δ7. 87-7.81(m,1H), 7.72-7.65(m,1H), 7.65-7.53(m,2H), 6.93-6.85(m,1H), 6.62-6.50(m,1H), 5.49-5.41(m ,1H),4.79–4.67(m,1H), 4.46(t,J=4.5Hz,2H), 3.71–3.63(m,2H), 3.60–3.47(m,1H), 3.47–3.34(m,1H ),3.04–2.92(m,2H),2.75–2.57(m,7H),2.04(s,3H),1.21 (s,3H),1.00(s,9H),0.96(s,3H),0.94( s, 3H), 0.81 (s, 3H). ESI-MS: m/z 760.6 [M+H] + .
实施例110Example 110
N-(2-(二甲基氨基)乙基)-3β-(2-(甲氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-51)N-(2-(dimethylamino)ethyl)-3β-(2-(methoxyacyl)benzoyl)oxy-oleanane-12-en-28-amide (compound B-51)
Figure PCTCN2020093248-appb-000148
Figure PCTCN2020093248-appb-000148
参照实施例106的方法，将N-乙酰乙醇胺替换成甲醇，N-(2-氨基乙基)吗啉替换成N,N-二甲基乙二胺，制得化合物B-51： 1H NMR(300MHz,CDCl 3)δ7.78–7.63(m,2H),7.58–7.48(m,2H),6.60–6.45(m,1H),5.45–5.30(m,1H),4.75(dd,J＝11.1,4.4Hz,1H),3.90(s,3H),3.44–3.30(m,1H),3.23–3.08(m,1H),2.60–2.48(m,1H),2.37(t,J＝5.9Hz,2H),2.23(s,6H),1.18(s,3H),0.97(s,6H),0.91(s,9H),0.81(s,3H).ESI-MS:m/z 689.5[M+H] +。 According to the method of Example 106, N-acetylethanolamine was replaced with methanol, and N-(2-aminoethyl)morpholine was replaced with N,N-dimethylethylenediamine to prepare compound B-51: 1 H NMR (300MHz, CDCl 3 )δ7.78–7.63(m,2H), 7.58–7.48(m,2H), 6.60–6.45(m,1H), 5.45–5.30(m,1H), 4.75(dd,J= 11.1,4.4Hz,1H),3.90(s,3H),3.44–3.30(m,1H), 3.23–3.08(m,1H), 2.60–2.48(m,1H), 2.37(t,J=5.9Hz ,2H),2.23(s,6H),1.18(s,3H),0.97(s,6H),0.91(s,9H),0.81(s,3H).ESI-MS:m/z 689.5(M+ H] + .
实施例111Example 111
N-(2-(二甲基氨基)乙基)-3β-(2-(异丙基氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-52)N-(2-(Dimethylamino)ethyl)-3β-(2-(isopropyloxyacyl)benzoyl)oxy-oleanorane-12-ene-28-amide (compound B- 52)
Figure PCTCN2020093248-appb-000149
Figure PCTCN2020093248-appb-000149
参照实施例106的方法，将N-乙酰乙醇胺替换成异丙醇，N-(2-氨基乙基)吗啉替换成N,N-二甲基乙二胺，制得化合物B-52： 1H NMR(300MHz,CDCl 3)δ7.81–7.65(m,2H),7.61–7.47(m,2H),6.66–6.47(m,1H),5.48–5.36(m,1H),5.35–5.22(m,1H),4.79(dd,J＝11.6,4.6Hz,1H),3.49–3.31(m,1H),3.29–3.09(m,1H),2.67–2.50(m,1H),2.40(t,J＝6.0Hz,2H),2.26(s,6H),1.39(d,6H),1.21(s,3H),1.01(s,6H),0.96(s,3H),0.95(s,6H),0.85(s,3H).ESI-MS:m/z 717.5[M+H] +。 Referring to the method of Example 106, N-acetylethanolamine was replaced with isopropanol, and N-(2-aminoethyl)morpholine was replaced with N,N-dimethylethylenediamine to obtain compound B-52: 1 H NMR (300MHz, CDCl 3 ) δ7.81-7.65(m,2H), 7.61-7.47(m,2H), 6.66-6.47(m,1H), 5.48-5.36(m,1H), 5.35-5.22( m,1H),4.79(dd,J=11.6,4.6Hz,1H),3.49–3.31(m,1H), 3.29–3.09(m,1H), 2.67–2.50(m,1H), 2.40(t, J = 6.0Hz, 2H), 2.26 (s, 6H), 1.39 (d, 6H), 1.21 (s, 3H), 1.01 (s, 6H), 0.96 (s, 3H), 0.95 (s, 6H), 0.85 (s, 3H). ESI-MS: m/z 717.5 [M+H] + .
实施例112Example 112
N-(2-(1-哌嗪基)乙基)-3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-53)N-(2-(1-piperazinyl)ethyl)-3β-(2-(2-acetamidoethoxyyl)benzoyl)oxy-oleanorane-12-ene-28-amide ( Compound B-53)
Figure PCTCN2020093248-appb-000150
Figure PCTCN2020093248-appb-000150
参照实施例106的方法，将N-(2-氨基乙基)吗啉替换成N-氨乙基哌嗪，制得化合物B-53： 1H NMR(300MHz,CDCl 3)δ7.85–7.77(m,1H),7.71–7.63(m,1H),7.64–7.49(m,2H),6.62–6.40(m,2H),5.47–5.31(m,1H),4.78–4.64(m,1H),4.44(t,J＝4.9Hz,2H),3.74–3.58(m,2H),3.52–3.39(m,1H),3.24–3.11(m,1H),2.99–2.87(m,3H),2.64–2.38(m,6H),2.31(2,3H),2.07–1.99(m,3H),1.18(s,3H),0.97(s,9H),0.92(s,6H),0.79(s,3H).ESI-MS:m/z 801.8[M+H] +。 According to the method of Example 106, N-(2-aminoethyl)morpholine was replaced with N-aminoethylpiperazine to prepare compound B-53: 1 H NMR (300MHz, CDCl 3 ) δ7.85-7.77 (m,1H), 7.71-7.63(m,1H), 7.64-7.49(m,2H), 6.62-6.40(m,2H), 5.47-5.31(m,1H), 4.78-4.64(m,1H) ,4.44(t,J=4.9Hz,2H),3.74–3.58(m,2H),3.52–3.39(m,1H),3.24–3.11(m,1H),2.99–2.87(m,3H),2.64 --2.38(m,6H),2.31(2,3H),2.07--1.99(m,3H),1.18(s,3H),0.97(s,9H),0.92(s,6H),0.79(s,3H) ). ESI-MS: m/z 801.8 [M+H] + .
实施例113Example 113
N-(2-(4-甲基-1-哌嗪基)乙基)-3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-54)N-(2-(4-Methyl-1-piperazinyl)ethyl)-3β-(2-(2-acetamidoethoxyyl)benzoyl)oxy-oleanorane-12-ene -28-amide (Compound B-54)
Figure PCTCN2020093248-appb-000151
Figure PCTCN2020093248-appb-000151
参照实施例106的方法，将N-(2-氨基乙基)吗啉替换成1-(2-氨乙基)-4-甲基哌嗪，制得化合物B-54： 1H NMR(300MHz,CDCl 3)δ7.78(d,J＝6.1Hz,1H),7.66(d,J＝7.9Hz,1H),7.53–7.42(m,2H),5.34–5.23(m,1H),4.73–4.66(m,1H),4.60–4.49(m,2H),3.52–2.98(m,6H),2.89–2.74(m,1H),1.15(s,3H),0.95(s,3H),0.92(s,9H),0.89(s,3H),0.72(s,3H).ESI-MS:m/z 816.7[M+H] +。 Referring to the method of Example 106, N-(2-aminoethyl)morpholine was replaced with 1-(2-aminoethyl)-4-methylpiperazine to obtain compound B-54: 1 H NMR (300MHz ,CDCl 3 )δ7.78(d,J=6.1Hz,1H),7.66(d,J=7.9Hz,1H),7.53–7.42(m,2H),5.34–5.23(m,1H),4.73– 4.66(m,1H), 4.60–4.49(m,2H), 3.52–2.98(m,6H), 2.89–2.74(m,1H), 1.15(s,3H), 0.95(s,3H), 0.92( s, 9H), 0.89 (s, 3H), 0.72 (s, 3H). ESI-MS: m/z 816.7 [M+H] + .
实施例114Example 114
N-(2-(三甲基铵基)乙基)-3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺溴化物(化合物B-55)N-(2-(Trimethylammonium)ethyl)-3β-(2-(2-Acetylaminoethoxyyl)benzoyl)oxy-oleanorane-12-ene-28-amide bromide Compound (Compound B-55)
Figure PCTCN2020093248-appb-000152
Figure PCTCN2020093248-appb-000152
参照实施例107的方法，将N-(2-氨基乙基)吗啉替换成化合物I-4，制得化合物B-55： 1H NMR(300MHz,methanol-d 4)δ7.84–7.74(m,2H),7.72–7.64(m,2H),5.45–5.39(m,1H),4.80–4.70(m,1H),4.40(t,J＝5.5Hz,2H),3.77–3.61(m,2H),3.58(t,J＝5.4Hz,2H),3.46(t,J＝6.9Hz,2H),3.24(s,9H),2.87(d,J＝9.7Hz,1H),2.25–2.11(m,1H),2.00(s,5H),1.26(s,3H),1.07(s,3H),1.02(s,3H),1.01(s,3H),0.98(s,6H),0.86(s,3H).ESI-MS:m/z 774.6[M-Br] +。 According to the method of Example 107, N-(2-aminoethyl)morpholine was replaced with compound I-4 to obtain compound B-55: 1 H NMR(300MHz, methanol-d 4 )δ7.84–7.74( m, 2H), 7.72–7.64(m, 2H), 5.45–5.39(m, 1H), 4.80–4.70(m, 1H), 4.40(t, J=5.5Hz, 2H), 3.77–3.61(m, 2H), 3.58 (t, J = 5.4 Hz, 2H), 3.46 (t, J = 6.9 Hz, 2H), 3.24 (s, 9H), 2.87 (d, J = 9.7 Hz, 1H), 2.25-2.11 ( m,1H),2.00(s,5H),1.26(s,3H),1.07(s,3H),1.02(s,3H),1.01(s,3H),0.98(s,6H),0.86(s , 3H). ESI-MS: m/z 774.6 [M-Br] + .
实施例115Example 115
N-(3-(二甲基氨基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-59)N-(3-(dimethylamino)propyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-12-en-28-amide (compound B-59)
Figure PCTCN2020093248-appb-000153
Figure PCTCN2020093248-appb-000153
参照实施例102的方法，将N-乙酰基乙二胺替换成3-二甲氨基丙胺，制得化合物B-59： 1H NMR(300MHz,CDCl 3)δ7.75–7.54(m,2H),7.52–7.30(m,2H),6.97–6.78(m,1H),5.45–5.29(m,1H),4.76–4.62(m,1H),3.92–3.78(m,1H),3.51–3.36(m,1H),3.31–3.14(m,1H),3.07–2.84(m,1H),2.72(s,6H),1.14(s,3H),0.98(s,3H),0.92(s,6H),0.88(s,6H),0.73(s,3H).ESI-MS:m/z 689.7[M+H] +。 According to the method of Example 102, N-acetylethylenediamine was replaced with 3-dimethylaminopropylamine to obtain compound B-59: 1 H NMR (300MHz, CDCl 3 )δ7.75–7.54(m,2H) ,7.52–7.30(m,2H), 6.97–6.78(m,1H), 5.45–5.29(m,1H), 4.76–4.62(m,1H), 3.92–3.78(m,1H), 3.51–3.36( m,1H),3.31--3.14(m,1H),3.07--2.84(m,1H),2.72(s,6H),1.14(s,3H),0.98(s,3H),0.92(s,6H) , 0.88 (s, 6H), 0.73 (s, 3H). ESI-MS: m/z 689.7 [M+H] + .
实施例116Example 116
N-(3-(4-吗啉基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-60)N-(3-(4-morpholinyl)propyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-12-en-28-amide (compound B-60)
Figure PCTCN2020093248-appb-000154
Figure PCTCN2020093248-appb-000154
参照实施例102的方法，将N-乙酰基乙二胺替换成N-(3-氨丙基)吗啉，制得化合物B-60： 1H NMR(300MHz,CDCl 3)δ7.82–7.67(m,1H),7.69–7.56(m,1H),7.55–7.36(m,2H),6.54–6.35(m,2H),5.51–5.24(m,1H),4.95–4.60(m,1H),4.04–3.77(m,4H),3.54–3.34(m,1H),3.24–3.03(m,1H),2.91–2.55(m,6H),1.15(s,3H),0.98(s,3H),0.91(s,3H),0.89(s,3H),0.87(s,3H),0.74(s,3H).ESI-MS:m/z 731.7[M+H] +。 Referring to the method of Example 102, N-acetylethylenediamine was replaced with N-(3-aminopropyl)morpholine to prepare compound B-60: 1 H NMR (300MHz, CDCl 3 ) δ7.82-7.67 (m,1H), 7.69-7.56(m,1H), 7.55-7.36(m,2H), 6.54-6.35(m,2H), 5.51-5.24(m,1H), 4.95-4.60(m,1H) ,4.04–3.77(m,4H),3.54–3.34(m,1H), 3.24–3.03(m,1H), 2.91–2.55(m,6H), 1.15(s,3H), 0.98(s,3H) , 0.91 (s, 3H), 0.89 (s, 3H), 0.87 (s, 3H), 0.74 (s, 3H). ESI-MS: m/z 731.7 [M+H] + .
实施例117Example 117
N-(3-(4-甲基-1-哌嗪基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-61)N-(3-(4-Methyl-1-piperazinyl)propyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-12-en-28-amide (compound B- 61)
Figure PCTCN2020093248-appb-000155
Figure PCTCN2020093248-appb-000155
参照实施例102的方法，将N-乙酰基乙二胺替换成1-(3-氨丙基)-4-甲基哌嗪，制得化合物B-61： 1H NMR(300MHz,CDCl 3)δ7.77–7.60(m,2H),7.60–7.37(m, 2H),6.60–6.19(m,3H),5.44–5.34(m,1H),4.79–4.57(m,1H),3.57–3.33(m,1H),3.27–3.04(m,8H),2.84–2.72(m,2H),1.19–1.15(m,3H),0.96(s,3H),0.92(s,3H),0.90(s,6H),0.74(s,3H).ESI-MS:m/z 744.7[M+H] +。 Referring to the method of Example 102, N-acetylethylenediamine was replaced with 1-(3-aminopropyl)-4-methylpiperazine to prepare compound B-61: 1 H NMR (300MHz, CDCl 3 ) δ7.77–7.60(m,2H), 7.60–7.37(m, 2H), 6.60–6.19(m,3H), 5.44–5.34(m,1H), 4.79–4.57(m,1H), 3.57–3.33 (m,1H), 3.27-3.04(m,8H), 2.84-2.72(m,2H), 1.19-1.15(m,3H), 0.96(s,3H), 0.92(s,3H), 0.90(s , 6H), 0.74 (s, 3H). ESI-MS: m/z 744.7 [M+H] + .
实施例118Example 118
N-(3-(1-四氢吡咯基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-62)N-(3-(1-tetrahydropyrrolyl)propyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-12-en-28-amide (compound B-62)
Figure PCTCN2020093248-appb-000156
Figure PCTCN2020093248-appb-000156
参照实施例102的方法，将N-乙酰基乙二胺替换成1-(3-氨基丙基)吡咯烷，制得化合物B-62： 1H NMR(300MHz,CDCl 3)δ7.70–7.54(m,2H),7.46–7.38(m,1H),7.36–7.30(m,1H),6.99–6.88(m,1H),5.39–5.27(m,1H),4.76–4.60(m,1H),3.52–3.37(m,1H),3.32–2.91(m,7H),2.81–2.65(m,1H),1.13(s,3H),0.99(s,3H),0.93(s,6H),0.88(s,6H),0.73(s,3H).ESI-MS:m/z 715.7[M+H] +。 According to the method of Example 102, N-acetylethylenediamine was replaced with 1-(3-aminopropyl)pyrrolidine to prepare compound B-62: 1 H NMR (300MHz, CDCl 3 ) δ 7.70-7.54 (m,2H),7.46--7.38(m,1H),7.36--7.30(m,1H),6.99-6.88(m,1H),5.39-5.27(m,1H),4.76-4.60(m,1H) ,3.52–3.37(m,1H), 3.32–2.91(m,7H), 2.81–2.65(m,1H), 1.13(s,3H), 0.99(s,3H), 0.93(s,6H), 0.88 (s, 6H), 0.73 (s, 3H). ESI-MS: m/z 715.7 [M+H] + .
实施例119Example 119
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-63)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-(2-carboxybenzoyl)oxy-oleanorane-12-en-28-amide (compound B-63)
Figure PCTCN2020093248-appb-000157
Figure PCTCN2020093248-appb-000157
参照实施例102的方法，将N-乙酰基乙二胺替换成4-(3-氨丙基)硫代吗啉-1,1-二氧化物，制得化合物B-63： 1H NMR(300MHz,CDCl 3)δ7.90–7.78(m,1H),7.75–7.65(m,1H),7.59–7.50(m,2H),6.17–6.02(m,1H),5.40–5.34(m,1H),5.32–5.24(m,1H),4.83–4.62(m,1H),3.53–3.33(m,1H),3.16–2.96(m,8H),2.62–2.42(m,3H),1.17(s,3H),0.96(s,3H),0.91(s,12H),0.76(s,3H).ESI-MS:m/z 779.7[M+H] +。 Referring to the method of Example 102, N-acetylethylenediamine was replaced with 4-(3-aminopropyl)thiomorpholine-1,1-dioxide to prepare compound B-63: 1 H NMR( 300MHz, CDCl 3 )δ7.90-7.78(m,1H), 7.75-7.65(m,1H), 7.59-7.50(m,2H), 6.17-6.02(m,1H), 5.40-5.34(m,1H) ), 5.32-5.24(m, 1H), 4.83-4.62(m, 1H), 3.53-3.33(m, 1H), 3.16-2.96(m, 8H), 2.62-2.42(m, 3H), 1.17(s , 3H), 0.96 (s, 3H), 0.91 (s, 12H), 0.76 (s, 3H). ESI-MS: m/z 779.7 [M+H] + .
实施例120Example 120
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-65)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-(2-(2-acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-12 -Ene-28-amide (compound B-65)
Figure PCTCN2020093248-appb-000158
Figure PCTCN2020093248-appb-000158
参照实施例106的方法，将N-(2-氨基乙基)吗啉替换成4-(3-氨丙基)硫代吗啉-1,1-二氧化物，制得化合物B-65： 1H NMR(300MHz,CDCl 3)δ7.84–7.77(m,1H),7.69–7.63(m,1H),7.62–7.50(m,2H),6.56–6.46(m,1H),6.05–5.96(m,1H),5.40–5.33(m,1H),4.74–4.65(m,1H),4.44(t,J＝4.9Hz,2H),3.69–3.58(m,2H),3.42(dd,J＝13.3,6.5Hz,1H),3.10–2.93(m,8H),2.58–2.45(m,3H),2.02(s,4H),1.19(s,3H),0.97(s,9H),0.91(s,6H),0.78(s,3H).ESI-MS:m/z 866.6[M+Na] +。 According to the method of Example 106, N-(2-aminoethyl)morpholine was replaced with 4-(3-aminopropyl)thiomorpholine-1,1-dioxide to obtain compound B-65: 1 H NMR(300MHz, CDCl 3 ) δ7.84-7.77(m,1H), 7.69-7.63(m,1H), 7.62-7.50(m,2H), 6.56-6.46(m,1H), 6.05-5.96 (m,1H),5.40–5.33(m,1H), 4.74–4.65(m,1H), 4.44(t,J=4.9Hz,2H), 3.69–3.58(m,2H),3.42(dd,J = 13.3, 6.5 Hz, 1H), 3.10–2.93 (m, 8H), 2.58–2.45 (m, 3H), 2.02 (s, 4H), 1.19 (s, 3H), 0.97 (s, 9H), 0.91 ( s, 6H), 0.78 (s, 3H). ESI-MS: m/z 866.6 [M+Na] + .
实施例121Example 121
N-(2-(1-哌啶基)乙基)-3β-(2-(甲氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-67)N-(2-(1-piperidinyl)ethyl)-3β-(2-(methoxyacyl)benzoyl)oxy-oleanane-12-en-28-amide (compound B-67 )
Figure PCTCN2020093248-appb-000159
Figure PCTCN2020093248-appb-000159
参照实施例106的方法，将N-乙酰乙醇胺替换成甲醇，N-(2-氨基乙基)吗啉替换成1-(2-氨乙基)哌啶，制得化合物B-67： 1H NMR(300MHz,CDCl 3)δ7.77–7.64(m,2H),7.58–7.48(m,2H),6.68–6.58(m,1H),5.42–5.34(m,1H),4.75(dd,J＝11.3,4.4Hz,1H),3.90(s,3H),3.46–3.32(m,1H),3.27–3.09(m,1H),2.59–2.48(m,1H),2.48–2.28(m,6H),1.18(s,3H),0.96(s,6H),0.92(s,9H),0.79(s,3H).ESI-MS:m/z 729.6[M+H] +。 According to the method of Example 106, N-acetylethanolamine was replaced with methanol, and N-(2-aminoethyl)morpholine was replaced with 1-(2-aminoethyl)piperidine to prepare compound B-67: 1 H NMR (300MHz, CDCl 3 ) δ7.77-7.64(m,2H), 7.58-7.48(m,2H), 6.68-6.58(m,1H), 5.42-5.34(m,1H), 4.75(dd,J =11.3,4.4Hz,1H),3.90(s,3H), 3.46–3.32(m,1H), 3.27–3.09(m,1H), 2.59–2.48(m,1H), 2.48–2.28(m,6H) ), 1.18 (s, 3H), 0.96 (s, 6H), 0.92 (s, 9H), 0.79 (s, 3H). ESI-MS: m/z 729.6 [M+H] + .
实施例122Example 122
N-(2-(1-哌啶基)乙基)-3β-(2-(异丙基氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-68)N-(2-(1-piperidinyl)ethyl)-3β-(2-(isopropyloxyacyl)benzoyl)oxy-oleanorane-12-ene-28-amide (compound B -68)
Figure PCTCN2020093248-appb-000160
Figure PCTCN2020093248-appb-000160
参照实施例106的方法，将N-乙酰乙醇胺替换成异丙醇，N-(2-氨基乙基)吗啉替换成1-(2-氨乙基)哌啶，制得化合物B-68： 1H NMR(300MHz,CDCl 3)δ7.76–7.62(m,2H),7.56–7.44(m,2H),6.67–6.57(m,1H),5.43–5.34(m,1H),5.32–5.16(m,1H),4.75(dd,J＝11.7,4.5Hz,1H),3.47–3.31(m,1H),3.25–3.10(m,1H),2.59–2.47(m,1H),2.47–2.27(m,6H),1.35(d,J＝6.2Hz,6H),1.17(s,3H),0.98(s,3H),0.96(s,3H),0.93(s,3H),0.92(s,6H),0.79(s,3H).ESI-MS:m/z 757.6[M+H] +。 According to the method of Example 106, N-acetylethanolamine was replaced with isopropanol, and N-(2-aminoethyl)morpholine was replaced with 1-(2-aminoethyl)piperidine to obtain compound B-68: 1 H NMR (300MHz, CDCl 3 ) δ7.76–7.62(m,2H), 7.56–7.44(m,2H), 6.67–6.57(m,1H), 5.43–5.34(m,1H), 5.32–5.16 (m,1H), 4.75(dd,J=11.7,4.5Hz,1H), 3.47–3.31(m,1H), 3.25–3.10(m,1H), 2.59–2.47(m,1H), 2.47–2.27 (m,6H),1.35(d,J=6.2Hz,6H),1.17(s,3H),0.98(s,3H),0.96(s,3H),0.93(s,3H),0.92(s, 6H), 0.79 (s, 3H). ESI-MS: m/z 757.6 [M+H] + .
实施例123Example 123
N-(2-(1-哌啶基)乙基)-3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-69)N-(2-(1-piperidinyl)ethyl)-3β-(2-(2-acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-12-ene-28-amide ( Compound B-69)
Figure PCTCN2020093248-appb-000161
Figure PCTCN2020093248-appb-000161
参照实施例106的方法将N-(2-氨基乙基)吗啉替换成1-(2-氨乙基)哌啶，制得化合物B-69： 1H NMR(300MHz,CDCl 3)δ7.81(d,J＝6.7Hz,1H),7.66(d,J＝6.6Hz,1H),7.56(p,J＝7.0Hz,2H),7.08–6.93(m,1H),6.59–6.46(m,1H),5.48(s,1H),4.71(t,J＝7.8Hz,1H),4.52–4.35(m,2H),3.72–3.35(m,5H),3.25–2.90(m,5H),2.69–2.56(m,1H),2.01(s,3H),1.18(s,3H),0.97(s,9H),0.94(s,3H),0.92(s,3H),0.77(s,3H).ESI-MS:m/z 800.7[M+H] +。 Referring to the method of Example 106, N-(2-aminoethyl)morpholine was replaced with 1-(2-aminoethyl)piperidine to obtain compound B-69: 1 H NMR (300MHz, CDCl 3 )δ7. 81(d,J=6.7Hz,1H),7.66(d,J=6.6Hz,1H),7.56(p,J=7.0Hz,2H),7.08–6.93(m,1H),6.59–6.46(m ,1H),5.48(s,1H),4.71(t,J=7.8Hz,1H),4.52–4.35(m,2H), 3.72–3.35(m,5H), 3.25–2.90(m,5H), 2.69--2.56(m,1H),2.01(s,3H),1.18(s,3H),0.97(s,9H),0.94(s,3H),0.92(s,3H),0.77(s,3H) .ESI-MS: m/z 800.7[M+H] + .
实施例124Example 124
N-(2-(1-四氢吡咯基)乙基)-3β-(2-(甲氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-70)N-(2-(1-Tetrahydropyrrolyl)ethyl)-3β-(2-(methoxyacyl)benzoyl)oxy-oleanorane-12-ene-28-amide (compound B- 70)
Figure PCTCN2020093248-appb-000162
Figure PCTCN2020093248-appb-000162
参照实施例106的方法，将N-乙酰乙醇胺替换成甲醇，N-(2-氨基乙基)吗啉替换成1-(2-氨乙基)吡咯烷，制得化合物B-70： 1H NMR(300MHz,CDCl 3)δ7.77–7.64(m,2H),7.58–7.47(m,2H),6.62–6.52(m,1H),5.40–5.28(m,1H),4.75(dd,J＝11.4,4.4Hz,1H),3.90(s,3H),3.50–3.38(m,1H),3.25–3.12(m,1H),2.68–2.41(m,7H),1.81–1.76(m,4H),1.18(s,3H),0.97(s,6H),0.91(s,9H),0.81(s,3H).ESI-MS:m/z 715.5[M+H] +。 According to the method of Example 106, N-acetylethanolamine was replaced with methanol, and N-(2-aminoethyl)morpholine was replaced with 1-(2-aminoethyl)pyrrolidine to obtain compound B-70: 1 H NMR (300MHz, CDCl 3 ) δ7.77-7.64(m,2H), 7.58-7.47(m,2H), 6.62-6.52(m,1H), 5.40-5.28(m,1H), 4.75(dd,J =11.4,4.4Hz,1H),3.90(s,3H),3.50–3.38(m,1H), 3.25–3.12(m,1H), 2.68–2.41(m,7H),1.81–1.76(m,4H) ), 1.18(s, 3H), 0.97(s, 6H), 0.91(s, 9H), 0.81(s, 3H). ESI-MS: m/z 715.5[M+H] + .
实施例125Example 125
N-(2-(1-四氢吡咯基)乙基)-3β-(2-(异丙基氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-71)N-(2-(1-Tetrahydropyrrolyl)ethyl)-3β-(2-(isopropyloxyacyl)benzoyl)oxy-oleanorane-12-ene-28-amide (compound B-71)
Figure PCTCN2020093248-appb-000163
Figure PCTCN2020093248-appb-000163
参照实施例106的方法，将N-乙酰乙醇胺替换成异丙醇，N-(2-氨基乙基)吗啉替换成1-(2-氨乙基)吡咯烷，制得化合物B-71： 1H NMR(300MHz,CDCl 3)δ7.81–7.60(m,2H),7.59–7.44(m,2H),6.64–6.46(m,1H),5.40–5.31(m,1H), 5.30–5.19(m,1H),4.75(dd,J＝11.3,4.1Hz,1H),3.53–3.32(m,1H),3.26–3.09(m,1H),2.69–2.35(m,7H),1.81–1.75(m,4H),1.18(s,3H),0.98(s,6H),0.93(s,3H),0.91(s,6H),0.81(s,3H).ESI-MS:m/z 743.6[M+H] +。 According to the method of Example 106, N-acetylethanolamine was replaced with isopropanol, and N-(2-aminoethyl)morpholine was replaced with 1-(2-aminoethyl)pyrrolidine to prepare compound B-71: 1 H NMR(300MHz, CDCl 3 ) δ7.81--7.60(m,2H), 7.59-7.44(m,2H), 6.64-6.46(m,1H), 5.40-5.31(m,1H), 5.30-5.19 (m,1H),4.75(dd,J=11.3,4.1Hz,1H),3.53–3.32(m,1H), 3.26–3.09(m,1H), 2.69–2.35(m,7H),1.81–1.75 (m,4H),1.18(s,3H),0.98(s,6H),0.93(s,3H),0.91(s,6H),0.81(s,3H).ESI-MS:m/z 743.6[ M+H] + .
实施例126Example 126
N-(2-(1-四氢吡咯基)乙基)-3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-72)N-(2-(1-Tetrahydropyrrolyl)ethyl)-3β-(2-(2-acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-12-en-28-amide (Compound B-72)
Figure PCTCN2020093248-appb-000164
Figure PCTCN2020093248-appb-000164
参照实施例106的方法，将N-(2-氨基乙基)吗啉替换成1-(2-氨乙基)吡咯烷，制得化合物B-72： 1H NMR(300MHz,CDCl 3)δ7.85–7.78(m,1H),7.70–7.63(m,1H),7.56(qd,J＝7.3,3.6Hz,2H),7.08–6.96(m,1H),6.64–6.48(m,1H),5.52–5.42(m,1H),4.78–4.65(m,1H),4.45(tq,J＝12.3,6.5,5.6Hz,2H),3.69–3.59(m,2H),3.58–3.34(m,6H),2.71–2.56(m,1H),2.01(s,3H),1.18(s,3H),0.99(s,3H),0.97(s,6H),0.94(s,3H),0.91(s,3H),0.76(s,3H).ESI-MS:m/z 786.7[M+H] +。 Referring to the method of Example 106, N-(2-aminoethyl)morpholine was replaced with 1-(2-aminoethyl)pyrrolidine to obtain compound B-72: 1 H NMR (300MHz, CDCl 3 )δ7 .85–7.78(m,1H),7.70–7.63(m,1H),7.56(qd,J=7.3,3.6Hz,2H),7.08–6.96(m,1H),6.64–6.48(m,1H) ,5.52–5.42(m,1H),4.78–4.65(m,1H), 4.45(tq,J=12.3,6.5,5.6Hz,2H), 3.69–3.59(m,2H),3.58–3.34(m, 6H), 2.71--2.56(m, 1H), 2.01(s, 3H), 1.18(s, 3H), 0.99(s, 3H), 0.97(s, 6H), 0.94(s, 3H), 0.91(s , 3H), 0.76 (s, 3H). ESI-MS: m/z 786.7 [M+H] + .
实施例127Example 127
N-(3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-吗啉(化合物B-73)N-(3β-(2-carboxybenzoyl)oxy-oleanorane-12-en-28-acyl)-morpholine (compound B-73)
Figure PCTCN2020093248-appb-000165
Figure PCTCN2020093248-appb-000165
参照实施例102的方法，将化合物XVI-1替换成化合物B-2，制得化合物B-73： 1H NMR(300MHz,DMSO-d 6)δ13.18(s,1H),7.69(s,1H),7.60(s,3H),5.08(s,1H),4.66–4.55(m,1H),3.50(s,8H),2.97–2.88(m,1H),1.10(s,3H),0.89(s,6H),0.85(s,6H),0.80(s,3H),0.65(s,3H).ESI-MS:m/z 672.5[M-H] -。 According to the method of Example 102, compound XVI-1 was replaced with compound B-2 to obtain compound B-73: 1 H NMR (300MHz, DMSO-d 6 ) δ 13.18 (s, 1H), 7.69 (s, 1H), 7.60(s, 3H), 5.08(s, 1H), 4.66-4.55(m, 1H), 3.50(s, 8H), 2.97-2.88(m, 1H), 1.10(s, 3H), 0.89 (s, 6H), 0.85 (s, 6H), 0.80 (s, 3H), 0.65 (s, 3H). ESI-MS: m/z 672.5[MH] - .
实施例128Example 128
N-(3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-硫代吗啉-1,1-二氧化物(化合物B-74)N-(3β-(2-carboxybenzoyl)oxy-oleanorane-12-ene-28-acyl)-thiomorpholine-1,1-dioxide (compound B-74)
Figure PCTCN2020093248-appb-000166
Figure PCTCN2020093248-appb-000166
参照实施例102的方法，将化合物XVI-1替换成化合物B-9，制得化合物B-74： 1H NMR(300MHz,DMSO-d 6)δ7.71(s,1H),7.58(s,3H),5.12(s,1H),4.62 (s,1H),3.94(s,4H),3.15–2.96(m,4H),2.92(s,1H),1.13(s,3H),0.92(s,3H),0.91(s,3H),0.88(s,3H),0.87(s,3H),0.82(s,3H),0.67(s,3H).m/z 720.4[M-H] -。 According to the method of Example 102, compound XVI-1 was replaced with compound B-9 to obtain compound B-74: 1 H NMR (300MHz, DMSO-d 6 ) δ 7.71 (s, 1H), 7.58 (s, 3H), 5.12 (s, 1H), 4.62 (s, 1H), 3.94 (s, 4H), 3.15-2.96 (m, 4H), 2.92 (s, 1H), 1.13 (s, 3H), 0.92 (s ,3H),0.91(s,3H),0.88(s,3H),0.87(s,3H),0.82(s,3H),0.67(s,3H).m/z 720.4[MH] - .
实施例129Example 129
N-(3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-四氢吡咯(化合物B-77)N-(3β-(2-carboxybenzoyl)oxy-oleanorane-12-en-28-acyl)-tetrahydropyrrole (compound B-77)
Figure PCTCN2020093248-appb-000167
Figure PCTCN2020093248-appb-000167
参照实施例102的方法，将化合物XVI-1替换成化合物B-26，制得化合物B-77： 1H NMR(300MHz,DMSO-d 6)δ13.20(s,1H),7.71(s,1H),7.63(s,3H),5.11(s,1H),4.62(t,J＝7.8Hz,1H),3.70–3.33(m,4H),3.08–2.98(m,1H),1.12(s,3H),0.90(s,6H),0.89(s,6H),0.82(s,3H),0.65(s,3H).ESI-MS:m/z 656.5[M-H] -。 According to the method of Example 102, compound XVI-1 was replaced with compound B-26 to obtain compound B-77: 1 H NMR (300MHz, DMSO-d 6 ) δ 13.20 (s, 1H), 7.71 (s, 1H), 7.63(s, 3H), 5.11(s, 1H), 4.62(t, J=7.8Hz, 1H), 3.70–3.33(m,4H), 3.08–2.98(m,1H), 1.12(s , 3H), 0.90 (s, 6H), 0.89 (s, 6H), 0.82 (s, 3H), 0.65 (s, 3H). ESI-MS: m/z 656.5 [MH] - .
实施例130Example 130
N-(3β-(2-甲氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-吗啉(化合物B-78)N-(3β-(2-methoxyacylbenzoyl)oxy-oleanane-12-en-28-acyl)-morpholine (compound B-78)
Figure PCTCN2020093248-appb-000168
Figure PCTCN2020093248-appb-000168
参照实施例106的方法，将N-乙酰乙醇胺替换成甲醇，N-(2-氨基乙基)吗啉替换成吗啉，制得化合物B-78： 1H NMR(300MHz,CDCl 3)δ7.77–7.65(m,2H),7.57–7.48(m,2H),5.34–5.22(m,1H),4.75(dd,J＝11.3,4.7Hz,1H),3.89(s,3H),3.76–3.54(m,8H),3.17–3.01(m,1H),1.16(s,3H),0.96(s,6H),0.94(s,3H),0.91(s,6H),0.75(s,3H).ESI-MS:m/z 688.5[M+H] +。 With reference to the method of Example 106, N-acetylethanolamine was replaced with methanol, and N-(2-aminoethyl)morpholine was replaced with morpholine to obtain compound B-78: 1 H NMR (300MHz, CDCl 3 )δ7. 77–7.65(m,2H), 7.57–7.48(m,2H), 5.34–5.22(m,1H), 4.75(dd,J=11.3,4.7Hz,1H), 3.89(s,3H), 3.76– 3.54(m,8H),3.17-3.01(m,1H),1.16(s,3H),0.96(s,6H),0.94(s,3H),0.91(s,6H),0.75(s,3H) .ESI-MS: m/z 688.5[M+H] + .
实施例131Example 131
N-(3β-(2-异丙基氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-吗啉(化合物B-79)N-(3β-(2-isopropyloxyacylbenzoyl)oxy-oleanorane-12-ene-28-acyl)-morpholine (compound B-79)
Figure PCTCN2020093248-appb-000169
Figure PCTCN2020093248-appb-000169
参照实施例106的方法，将N-乙酰乙醇胺替换成异丙醇，N-(2-氨基乙基)吗啉替换成吗啉，制得化合物B-79： 1H NMR(300MHz,CDCl 3)δ7.77–7.60(m,2H),7.58–7.42(m,2H),5.37–5.15(m,2H),4.75(dd,J＝11.6,4.7Hz,1H),3.84– 3.42(m,8H),3.18–3.01(m,1H),1.35(d,J＝6.1Hz,6H),1.16(s,3H),0.97(s,3H),0.96(s,3H),0.94(s,3H),0.93(s,3H),0.91(s,3H),0.76(s,3H).ESI-MS:m/z 738.5[M+Na] +。 According to the method of Example 106, N-acetylethanolamine was replaced with isopropanol, and N-(2-aminoethyl)morpholine was replaced with morpholine to obtain compound B-79: 1 H NMR (300MHz, CDCl 3 ) δ7.77–7.60 (m, 2H), 7.58–7.42 (m, 2H), 5.37–5.15 (m, 2H), 4.75 (dd, J = 11.6, 4.7 Hz, 1H), 3.84– 3.42 (m, 8H) ), 3.18–3.01(m,1H),1.35(d,J=6.1Hz,6H),1.16(s,3H),0.97(s,3H),0.96(s,3H),0.94(s,3H) , 0.93 (s, 3H), 0.91 (s, 3H), 0.76 (s, 3H). ESI-MS: m/z 738.5 [M+Na] + .
实施例132Example 132
N-(3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-吗啉(化合物B-80)N-(3β-(2-(2-Acetylamino)ethoxyacylbenzoyl)oxy-oleanane-12-ene-28-acyl)-morpholine (compound B-80)
Figure PCTCN2020093248-appb-000170
Figure PCTCN2020093248-appb-000170
参照实施例106的方法，将N-(2-氨基乙基)吗啉替换成吗啉，制得化合物B-80： 1H NMR(300MHz,CDCl 3)δ7.87–7.75(m,1H),7.72–7.61(m,1H),7.61–7.48(m,2H),6.64–6.50(m,1H),5.34–5.24(m,1H),4.80–4.63(m,1H),4.53–4.33(m,2H),3.80–3.41(m,9H),3.20–2.99(m,1H),2.32–2.08(m,1H),2.05(s,3H),1.16(s,3H),0.97(s,9H),0.94(s,3H),0.91(s,3H),0.76(s,3H).ESI-MS:m/z781.7[M+Na] +。 According to the method of Example 106, N-(2-aminoethyl)morpholine was replaced with morpholine to obtain compound B-80: 1 H NMR (300MHz, CDCl 3 ) δ 7.87-7.75 (m, 1H) ,7.72–7.61(m,1H), 7.61–7.48(m,2H), 6.64–6.50(m,1H), 5.34–5.24(m,1H), 4.80–4.63(m,1H), 4.53–4.33( m, 2H), 3.80--3.41(m, 9H), 3.20--2.99(m, 1H), 2.32--2.08(m, 1H), 2.05(s, 3H), 1.16(s, 3H), 0.97(s, 9H), 0.94 (s, 3H), 0.91 (s, 3H), 0.76 (s, 3H). ESI-MS: m/z 781.7 [M+Na] + .
实施例133Example 133
3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酸2-(1-哌啶基)乙酯(化合物B-87)3β-(2-Carboxybenzoyl)oxy-oleanolic-12-en-28-acid 2-(1-piperidinyl)ethyl ester (Compound B-87)
Figure PCTCN2020093248-appb-000171
Figure PCTCN2020093248-appb-000171
参照实施例102的方法，将化合物XVI-1替换成化合物B-31，制得化合物B-87： 1H NMR(300MHz,CDCl 3)δ7.78(d,J＝6.1Hz,1H),7.66(d,J＝7.9Hz,1H),7.53–7.42(m,2H),5.34–5.23(m,1H),4.73–4.66(m,1H),4.60–4.49(m,2H),3.52–2.98(m,6H),2.89–2.74(m,1H),1.15(s,3H),0.95(s,3H),0.92(s,9H),0.89(s,3H),0.72(s,3H).ESI-MS:m/z 717.6[M+H] +。 According to the method of Example 102, compound XVI-1 was replaced with compound B-31 to obtain compound B-87: 1 H NMR (300MHz, CDCl 3 ) δ 7.78 (d, J = 6.1 Hz, 1H), 7.66 (d,J=7.9Hz,1H), 7.53–7.42(m,2H), 5.34–5.23(m,1H), 4.73–4.66(m,1H), 4.60–4.49(m,2H), 3.52–2.98 (m, 6H), 2.89-2.74 (m, 1H), 1.15 (s, 3H), 0.95 (s, 3H), 0.92 (s, 9H), 0.89 (s, 3H), 0.72 (s, 3H). ESI-MS: m/z 717.6 [M+H] + .
实施例134Example 134
3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酸2-(4-吗啉基)乙酯(化合物B-88)3β-(2-Carboxybenzoyl)oxy-oleanorane-12-ene-28-acid 2-(4-morpholinyl)ethyl ester (Compound B-88)
Figure PCTCN2020093248-appb-000172
Figure PCTCN2020093248-appb-000172
参照实施例102的方法，将化合物XVI-1替换成化合物B-32，制得化合物B-88： 1H NMR(300MHz,CDCl 3)δ7.88–7.79(m,1H),7.71–7.63(m,1H),7.56–7.46(m,2H),5.31–5.24(m,1H),4.76(dd,J＝11.5,4.3Hz,1H),4.35–4.22(m,2H),3.88–3.75(m,4H),2.91–2.78(m,3H),2.77–2.62(m,4H),1.15(s,3H),0.97(s,3H),0.93(s,3H),0.92(s,3H),0.91(s,3H),0.90(s,3H),0.73(s,3H).ESI-MS:m/z 716.5[M+H] +。 According to the method of Example 102, compound XVI-1 was replaced with compound B-32 to obtain compound B-88: 1 H NMR (300MHz, CDCl 3 )δ7.88-7.79(m,1H), 7.71-7.63( m,1H),7.56–7.46(m,2H),5.31–5.24(m,1H), 4.76(dd,J=11.5,4.3Hz,1H), 4.35–4.22(m,2H), 3.88–3.75( m,4H), 2.91–2.78(m,3H), 2.77–2.62(m,4H), 1.15(s,3H), 0.97(s,3H), 0.93(s,3H), 0.92(s,3H) , 0.91 (s, 3H), 0.90 (s, 3H), 0.73 (s, 3H). ESI-MS: m/z 716.5 [M+H] + .
实施例135Example 135
3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酸2-(1-四氢吡咯基)乙酯(化合物B-89)3β-(2-Carboxybenzoyl)oxy-oleanolic-12-ene-28-acid 2-(1-tetrahydropyrrolyl)ethyl ester (Compound B-89)
Figure PCTCN2020093248-appb-000173
Figure PCTCN2020093248-appb-000173
参照实施例102的方法，将化合物XVI-1替换成化合物B-33，制得化合物B-89： 1H NMR(300MHz,CDCl 3)δ7.76(d,J＝7.3Hz,1H),7.59(d,J＝7.5Hz,1H),7.50–7.35(m,2H),5.33–5.25(m,1H),4.79–4.67(m,1H),4.52–4.42(m,2H),3.34–3.11(m,6H),2.91–2.76(m,1H),1.15(s,4H),0.98(s,2H),0.96–0.90(m,9H),0.89(s,3H),0.72(s,3H).ESI-MS:m/z 702.5[M+H] +。 According to the method of Example 102, compound XVI-1 was replaced with compound B-33 to obtain compound B-89: 1 H NMR (300MHz, CDCl 3 ) δ 7.76 (d, J = 7.3 Hz, 1H), 7.59 (d,J=7.5Hz,1H),7.50–7.35(m,2H),5.33–5.25(m,1H),4.79–4.67(m,1H),4.52–4.42(m,2H),3.34–3.11 (m,6H),2.91--2.76(m,1H),1.15(s,4H),0.98(s,2H),0.96--0.90(m,9H),0.89(s,3H),0.72(s,3H) ). ESI-MS: m/z 702.5 [M+H] + .
实施例136Example 136
3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酸2-(4-吗啉基)乙酯(化合物B-93)3β-(2-(2-Acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-12-ene-28-acid 2-(4-morpholinyl)ethyl ester (Compound B-93)
Figure PCTCN2020093248-appb-000174
Figure PCTCN2020093248-appb-000174
参照实施例106的方法制得化合物XVII-4，取化合物XVII-4(300mg，0.435mmol)溶于N,N-二甲基甲酰胺(8mL)中，依次加入1,2-二溴乙烷(375μL，4.348mmol)、碳酸钾(60mg，0.435mmol)，将反应液升温至50℃，搅拌反应3小时。TLC检测反应完全后，用二氯甲烷(8mL)稀释反应液，水洗(8mL)反应液，水层用二氯甲烷(8mL)萃取，合并有机相，水(8mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝2：1)纯化，得化合物XVIII-1(白色固体，331mg，产率96％)。The compound XVII-4 was prepared by referring to the method of Example 106. The compound XVII-4 (300 mg, 0.435 mmol) was dissolved in N,N-dimethylformamide (8 mL), and 1,2-dibromoethane was sequentially added. (375 μL, 4.348 mmol), potassium carbonate (60 mg, 0.435 mmol), the reaction solution was heated to 50° C., and the reaction was stirred for 3 hours. After the completion of the reaction was detected by TLC, the reaction solution was diluted with dichloromethane (8mL), washed with water (8mL), the aqueous layer was extracted with dichloromethane (8mL), the organic phases were combined, washed with water (8mL×2), and anhydrous sulfuric acid Dry with sodium, filter, concentrate the filtrate, and purify by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound XVIII-1 (white solid, 331 mg, yield 96%).
取化合物XVIII-1(200mg，0.251mmol)溶于无水N,N-二甲基甲酰胺(10mL)中，依次加入吗啉(45μL，0.52mmol)和碳酸钾(48mg，0.35mmol)，将反应液升温至50℃，搅拌反应。TLC检测反应完全后，用二氯甲烷(10mL)和水(5mL)处理反应液，水层用二氯甲烷(10mL×2)萃取，合并有机相，水(10mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝30：1)纯化，得化合物B-93(白色固体，97mg，产率48％)： 1H NMR(300MHz,CDCl 3)δ7.88–7.76(m,1H),7.71–7.63(m,1H),7.62–7.49(m,2H),6.62–6.51(m,1H),5.37–5.18(m,1H),4.86–4.59(m,1H),4.52–4.36(m,2H),4.31–4.08(m,2H),3.82–3.58(m,6H),2.99–2.81(m,1H),2.71–2.58(m,2H),2.58–2.44(m,3H),2.02(s,3H),1.16(s,3H),0.97(s,9H),0.93(s,3H),0.91(s,3H),0.76(s,3H).ESI-MS:m/z 803.8[M+H] +。 Dissolve compound XVIII-1 (200mg, 0.251mmol) in anhydrous N,N-dimethylformamide (10mL), add morpholine (45μL, 0.52mmol) and potassium carbonate (48mg, 0.35mmol) in sequence, The reaction solution was heated to 50°C, and the reaction was stirred. After the completion of the reaction was detected by TLC, the reaction solution was treated with dichloromethane (10mL) and water (5mL), the aqueous layer was extracted with dichloromethane (10mL×2), the organic phases were combined, washed with water (10mL×2), and anhydrous sulfuric acid The sodium was dried, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 30:1) to obtain compound B-93 (white solid, 97 mg, yield 48%): 1 H NMR (300MHz, CDCl 3 )δ7.88–7.76(m,1H), 7.71–7.63(m,1H), 7.62–7.49(m,2H), 6.62–6.51(m,1H), 5.37–5.18(m,1H), 4.86 --4.59 (m, 1H), 4.52 - 4.36 (m, 2H), 4.31 - 4.08 (m, 2H), 3.82 - 3.58 (m, 6H), 2.99 - 2.81 (m, 1H), 2.71 - 2.58 (m, 2H),2.58--2.44(m,3H),2.02(s,3H),1.16(s,3H),0.97(s,9H),0.93(s,3H),0.91(s,3H),0.76(s , 3H). ESI-MS: m/z 803.8 [M+H] + .
实施例137Example 137
3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酸2-三甲基铵基乙酯溴化物(化合物B-94)3β-(2-(2-Acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-12-ene-28-acid 2-trimethylammonium ethyl bromide (Compound B-94)
Figure PCTCN2020093248-appb-000175
Figure PCTCN2020093248-appb-000175
参照实施例136的方法，将吗啉替换成三甲胺的四氢呋喃溶液，制得化合物B-94： 1H NMR(300MHz,methanol-d 6)δ7.84–7.74(m,2H),7.71–7.63(m,2H),5.37–5.31(m,1H),4.79–4.70(m,1H),4.61–4.49(m,2H),4.41(t,J＝5.4Hz,2H),3.86–3.71(m,2H),3.58(t,J＝5.4Hz,2H),3.29(s,9H),2.92(d,J＝10.1Hz,1H),2.23–2.10(m,1H),2.00(s,5H),1.26(s,3H),1.07(s,3H),1.03(s,3H),1.00(s,3H),0.99(s,6H),0.85(s,3H).ESI-MS:m/z 775.7[M-Br] +。 According to the method of Example 136, morpholine was replaced with trimethylamine in tetrahydrofuran to obtain compound B-94: 1 H NMR(300MHz, methanol-d 6 )δ7.84–7.74(m,2H), 7.71–7.63 (m,2H),5.37–5.31(m,1H), 4.79–4.70(m,1H), 4.61–4.49(m,2H), 4.41(t,J=5.4Hz,2H), 3.86–3.71(m ,2H),3.58(t,J=5.4Hz,2H), 3.29(s,9H), 2.92(d,J=10.1Hz,1H), 2.23-2.10(m,1H),2.00(s,5H) ,1.26(s,3H),1.07(s,3H),1.03(s,3H),1.00(s,3H),0.99(s,6H),0.85(s,3H).ESI-MS: m/z 775.7[M-Br] + .
实施例138Example 138
3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酸3-三甲基铵基丙酯溴化物(化合物B-95)3β-(2-(2-Acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-12-ene-28-acid 3-trimethylammonium propyl bromide (Compound B-95)
Figure PCTCN2020093248-appb-000176
Figure PCTCN2020093248-appb-000176
参照实施例136的方法，将1,2-二溴乙烷替换成1,3-二溴丙烷，吗啉替换成三甲胺，制得化合物B-95： 1H NMR(300MHz,methanol-d 6)δ7.84–7.75(m,2H),7.72–7.63(m,2H),5.39–5.33(m,1H),4.76(dd,J＝10.2,5.8Hz,1H),4.41(t,J＝5.5Hz,2H),4.25–4.12(m,2H),3.58(t,J＝5.5Hz,2H),3.48(dd,J＝11.0,5.7Hz,2H),3.23(s,9H),2.99–2.89(m,1H),2.28–2.16(m,2H),2.16–2.06(m,1H),2.00(s,5H),1.26(s,3H),1.07(s,3H),1.02(s,3H),1.01(s,3H),0.99(s,6H),0.85(s,3H).ESI-MS:m/z 789.6[M-Br] +。 With reference to the method in Example 136, 1,2-dibromoethane was replaced with 1,3-dibromopropane, and morpholine was replaced with trimethylamine to obtain compound B-95: 1 H NMR (300MHz, methanol-d 6 )δ7.84–7.75(m,2H),7.72–7.63(m,2H),5.39–5.33(m,1H), 4.76(dd,J＝10.2,5.8Hz,1H), 4.41(t,J＝ 5.5Hz, 2H), 4.25-4.12 (m, 2H), 3.58 (t, J = 5.5 Hz, 2H), 3.48 (dd, J = 11.0, 5.7 Hz, 2H), 3.23 (s, 9H), 2.99- 2.89(m,1H), 2.28–2.16(m,2H), 2.16–2.06(m,1H), 2.00(s,5H), 1.26(s,3H), 1.07(s,3H), 1.02(s, 3H), 1.01 (s, 3H), 0.99 (s, 6H), 0.85 (s, 3H). ESI-MS: m/z 789.6 [M-Br] + .
实施例139Example 139
3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酸2-(4-甲基-1-哌嗪基)乙酯(化合物B-97)3β-(2-(2-Acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-12-ene-28-acid 2-(4-methyl-1-piperazinyl)ethyl ester ( Compound B-97)
Figure PCTCN2020093248-appb-000177
Figure PCTCN2020093248-appb-000177
参照实施例136的方法，将吗啉替换成N-甲基哌嗪，制得化合物B-97： 1H NMR(300MHz,CDCl 3)δ7.85–7.78(m,1H),7.69–7.63(m,1H),7.62–7.50(m,2H),6.59–6.48(m,1H),5.30–5.23(m,1H),4.77–4.63(m,1H),4.51–4.37(m,2H),4.28–4.04(m,2H),3.73–3.57(m,2H),2.93–2.80(m,1H),2.73–2.38(m,8H),2.34–2.26(m,2H),2.02(s,3H),1.15(s,3H),0.97(s,9H),0.92(s,3H),0.91(s,3H),0.75(s,3H).ESI-MS:m/z 816.8[M+H] +。 According to the method of Example 136, morpholine was replaced with N-methylpiperazine to obtain compound B-97: 1 H NMR (300MHz, CDCl 3 )δ7.85-7.78(m,1H),7.69-7.63( m,1H), 7.62–7.50(m,2H), 6.59–6.48(m,1H), 5.30–5.23(m,1H), 4.77–4.63(m,1H), 4.51–4.37(m,2H), 4.28–4.04(m,2H), 3.73–3.57(m,2H), 2.93–2.80(m,1H), 2.73–2.38(m,8H), 2.34–2.26(m,2H), 2.02(s,3H) ),1.15(s,3H),0.97(s,9H),0.92(s,3H),0.91(s,3H),0.75(s,3H).ESI-MS:m/z 816.8[M+H] + .
实施例140Example 140
N-(2-(乙酰氨基)乙基)-3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-105)N-(2-(Acetylamino)ethyl)-3β-(2-(2-Acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-12-ene-28-amide (compound B- 105)
Figure PCTCN2020093248-appb-000178
Figure PCTCN2020093248-appb-000178
参照实施例106的方法，将N-(2-氨基乙基)吗啉替换成N-乙酰基乙二胺，制得化合物B-105： 1H NMR(300MHz,CDCl 3)δ7.86–7.78(m,1H),7.70–7.63(m,1H),7.57(qt,J＝7.3,3.6Hz,2H),6.68–6.52(m,2H),6.42(t,J＝4.8Hz,1H),5.42(s,1H),4.77–4.64(m,1H),4.44(t,J＝4.8Hz,2H),3.71–3.57(m,2H),3.54– 3.16(m,4H),2.62–2.51(m,1H),2.02(s,3H),1.97(s,3H),1.18(s,3H),0.97(s,9H),0.92(s,6H),0.77(s,3H).ESI-MS:m/z 796.7[M+Na] +。 According to the method of Example 106, N-(2-aminoethyl)morpholine was replaced with N-acetylethylenediamine to prepare compound B-105: 1 H NMR (300MHz, CDCl 3 ) δ7.86–7.78 (m,1H),7.70–7.63(m,1H),7.57(qt,J=7.3,3.6Hz,2H),6.68–6.52(m,2H),6.42(t,J=4.8Hz,1H), 5.42(s,1H),4.77–4.64(m,1H), 4.44(t,J=4.8Hz,2H), 3.71–3.57(m,2H),3.54– 3.16(m,4H), 2.62–2.51( m,1H),2.02(s,3H),1.97(s,3H),1.18(s,3H),0.97(s,9H),0.92(s,6H),0.77(s,3H).ESI-MS :m/z 796.7[M+Na] + .
实施例141Example 141
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-吖丁啶(化合物B-106)N-(3β-Hydroxy-oleanan-12-ene-28-acyl)-azetidine (Compound B-106)
Figure PCTCN2020093248-appb-000179
Figure PCTCN2020093248-appb-000179
参照实施例73的方法，将N,N-二甲基乙二胺替换成杂氮环丁烷，制得化合物B-106： 1H NMR(300MHz,CDCl 3)δ5.24(t,J＝3.2Hz,1H),4.50–3.83(m,4H),3.29–3.12(m,1H),2.85(dd,J＝13.4,3.5Hz,1H),2.21(t d,J＝7.5Hz,2H),1.12(s,3H),0.98(s,3H),0.92(s,3H),0.91(s,3H),0.89(s,3H),0.78(s,3H),0.77(s,3H).ESI-MS:m/z 518.4[M+Na] +。 With reference to the method of Example 73, N,N-dimethylethylenediamine was replaced with azacyclobutane to obtain compound B-106: 1 H NMR (300MHz, CDCl 3 )δ5.24(t,J= 3.2Hz,1H), 4.50–3.83(m,4H), 3.29–3.12(m,1H), 2.85(dd,J=13.4,3.5Hz,1H),2.21(td,J=7.5Hz,2H), 1.12(s,3H),0.98(s,3H),0.92(s,3H),0.91(s,3H),0.89(s,3H),0.78(s,3H),0.77(s,3H).ESI -MS: m/z 518.4 [M+Na] + .
实施例142Example 142
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-3-羟基-吖丁啶(化合物B-107)N-(3β-Hydroxy-oleanane-12-ene-28-acyl)-3-hydroxy-azetidine (Compound B-107)
Figure PCTCN2020093248-appb-000180
Figure PCTCN2020093248-appb-000180
参照实施例73的方法，将N,N-二甲基乙二胺替换成氮杂环丁烷-3-醇，制得化合物B-107： 1H NMR(300MHz,CDCl 3)δ5.25(m,1H),4.68–4.53(m,1H),4.49–4.20(m,2H),4.14–3.77(m,2H),3.28–3.12(m,1H),2.87–2.77(m,1H),1.13(s,3H),0.98(s,3H),0.92(s,3H),0.90(s,6H),0.78(s,3H),0.75(s,3H).ESI-MS:m/z 510.4[M-H] -。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with azetidine-3-ol to obtain compound B-107: 1 H NMR (300MHz, CDCl 3 )δ 5.25( m,1H), 4.68--4.53(m,1H), 4.49--4.20(m,2H), 4.14--3.77(m,2H), 3.28--3.12(m,1H), 2.87--2.77(m,1H), 1.13(s,3H),0.98(s,3H),0.92(s,3H),0.90(s,6H),0.78(s,3H),0.75(s,3H).ESI-MS: m/z 510.4 [MH] - .
实施例143Example 143
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-3-甲氧基-吖丁啶(化合物B-110)N-(3β-Hydroxy-oleanane-12-ene-28-acyl)-3-methoxy-azetidine (Compound B-110)
Figure PCTCN2020093248-appb-000181
Figure PCTCN2020093248-appb-000181
参照实施例73的方法，将N,N-二甲基乙二胺替换成3-甲氧基氮杂丁烷，制得化合物B-110： 1H NMR(300MHz,CDCl 3)δ5.28–5.21(m,1H),4.55–3.75(m,5H),3.28(s,3H),3.25–3.13(m,1H),2.90–2.78(m,1H),1.12(s,3H),0.98(s,3H),0.92(s,3H),0.90(s,3H),0.89(s,3H),0.78(s,3H),0.75(s,3H).ESI-MS:m/z 548.5[M+Na] +。 With reference to the method of Example 73, N,N-dimethylethylenediamine was replaced with 3-methoxyazetidine to obtain compound B-110: 1 H NMR (300MHz, CDCl 3 ) δ 5.28- 5.21(m,1H),4.55-3.75(m,5H), 3.28(s,3H), 3.25-3.13(m,1H), 2.90-2.78(m,1H), 1.12(s,3H), 0.98( s,3H),0.92(s,3H),0.90(s,3H),0.89(s,3H),0.78(s,3H),0.75(s,3H).ESI-MS:m/z 548.5(M +Na] + .
实施例144Example 144
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-3-羧基-吖丁啶(化合物B-111)N-(3β-Hydroxy-oleanan-12-ene-28-acyl)-3-carboxy-azetidine (Compound B-111)
Figure PCTCN2020093248-appb-000182
Figure PCTCN2020093248-appb-000182
参照实施例86的方法，将L-脯氨酸苄酯盐酸盐替换成化合物VIII-3，制得化合物B-111： 1H NMR(300MHz,CDCl 3)δ5.34–5.16(m,1H),4.71–4.02(m,4H),3.40–3.28(m,1H),3.28–3.17(m,1H),2.94–2.71(m,1H),1.13(s,3H),0.99(s,3H),0.92(s,3H),0.90(s,6H),0.79(s,3H),0.75(s,3H).ESI-MS:m/z 538.4[M-H] -。 According to the method of Example 86, the L-proline benzyl ester hydrochloride was replaced with compound VIII-3 to obtain compound B-111: 1 H NMR (300MHz, CDCl 3 )δ5.34–5.16 (m, 1H ), 4.71–4.02(m,4H), 3.40–3.28(m,1H), 3.28–3.17(m,1H), 2.94–2.71(m,1H), 1.13(s,3H), 0.99(s,3H) ), 0.92(s, 3H), 0.90(s, 6H), 0.79(s, 3H), 0.75(s, 3H). ESI-MS: m/z 538.4[MH] - .
实施例145Example 145
N-(2-(1,1-二氧代硫代吗啉基)乙基)-3β-(2-(2-乙酰氨基)乙氧酰基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-118)N-(2-(1,1-dioxothiomorpholinyl)ethyl)-3β-(2-(2-acetylamino)ethoxyacylbenzoyl)oxy-oleanorane-12 -Ene-28-amide (compound B-118)
Figure PCTCN2020093248-appb-000183
Figure PCTCN2020093248-appb-000183
参照实施例106的方法，将N-(2-氨基乙基)吗啉替换成4-(2-氨乙基)硫代吗啉-1,1-二氧化物，制得化合物B-118： 1H NMR(300MHz,CDCl 3)δ7.81(d,J＝6.9Hz,1H),7.66(d,J＝6.7Hz,1H),7.62–7.51(m,2H),6.58–6.47(m,1H),6.27–6.12(m,1H),5.40–5.33(m,1H),4.77–4.66(m,1H),4.48–4.38(m,2H),3.69–3.60(m,2H),3.56–3.43(m,1H),3.23–3.01(m,9H),2.69–2.58(m,2H),2.57–2.45(m,1H),1.19(s,3H),0.98(s,9H),0.92(s,3H),0.91(s,3H),0.79(s,3H).ESI-MS:m/z 872.7[M+Na] +。 Referring to the method of Example 106, N-(2-aminoethyl)morpholine was replaced with 4-(2-aminoethyl)thiomorpholine-1,1-dioxide to obtain compound B-118: 1 H NMR(300MHz,CDCl 3 )δ7.81(d,J=6.9Hz,1H), 7.66(d,J=6.7Hz,1H), 7.62–7.51(m,2H), 6.58–6.47(m, 1H), 6.27–6.12(m,1H), 5.40–5.33(m,1H), 4.77–4.66(m,1H), 4.48–4.38(m,2H), 3.69–3.60(m,2H), 3.56– 3.43(m,1H), 3.23-3.01(m,9H), 2.69-2.58(m,2H), 2.57-2.45(m,1H), 1.19(s,3H), 0.98(s,9H), 0.92( s, 3H), 0.91 (s, 3H), 0.79 (s, 3H). ESI-MS: m/z 872.7 [M+Na] + .
实施例146Example 146
N-(3-(1-哌啶基)丙基)-3β-(2-羧基苯甲酰基)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-119)N-(3-(1-piperidinyl)propyl)-3β-(2-carboxybenzoyl)oxy-oleanane-12-en-28-amide (compound B-119)
Figure PCTCN2020093248-appb-000184
Figure PCTCN2020093248-appb-000184
参照实施例102的方法，将N-乙酰基乙二胺替换成1-(3-氨基丙基)哌啶，制得化合物B-119： 1H NMR(300MHz,DMSO-d 6)δ7.70(s,1H),7.55(s,2H),7.31(s,1H),5.22(s,1H),4.61(s,1H),3.30(s,3H),3.04(s,3H),2.80(s,2H),1.12(s,3H),1.05(s,3H),0.92(s,3H),0.90(s,3H),0.88(s,3H),0.82(s,3H),0.68(s,3H). ESI-MS:m/z 727.6[M-H] -。 According to the method of Example 102, N-acetylethylenediamine was replaced with 1-(3-aminopropyl)piperidine to prepare compound B-119: 1 H NMR (300MHz, DMSO-d 6 ) δ 7.70 (s, 1H), 7.55(s, 2H), 7.31(s, 1H), 5.22(s, 1H), 4.61(s, 1H), 3.30(s, 3H), 3.04(s, 3H), 2.80( s,2H),1.12(s,3H),1.05(s,3H),0.92(s,3H),0.90(s,3H),0.88(s,3H),0.82(s,3H),0.68(s ,3H). ESI-MS: m/z 727.6[MH] - .
实施例147Example 147
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-(2R)-羧基-四氢吡咯(化合物B-120)N-(3β-Hydroxy-oleanorane-12-ene-28-acyl)-(2R)-carboxy-tetrahydropyrrole (Compound B-120)
Figure PCTCN2020093248-appb-000185
Figure PCTCN2020093248-appb-000185
参照实施例86的方法，将L-脯氨酸苄酯盐酸盐替换成D-脯氨酸苄酯盐酸盐，制得化合物B-120： 1H NMR(300MHz,DMSO-d 6)δ5.26(m,1H),4.64–4.56(m,1H),3.84–3.73(m,1H),3.73–3.61(m,1H),3.26–3.15(m,1H),3.12–2.98(m,1H),2.41–2.26(m,1H),1.15(s,3H),0.98(s,3H),0.93(s,3H),0.91(s,3H),0.89(s,3H),0.77(s,3H),0.72(s,3H).ESI-MS:m/z 552.4[M-H] -。 According to the method of Example 86, L-proline benzyl ester hydrochloride was replaced with D-proline benzyl ester hydrochloride to prepare compound B-120: 1 H NMR (300MHz, DMSO-d 6 )δ5 .26(m,1H), 4.64–4.56(m,1H), 3.84–3.73(m,1H), 3.73–3.61(m,1H), 3.26–3.15(m,1H), 3.12–2.98(m, 1H),2.41-2.26(m,1H),1.15(s,3H),0.98(s,3H),0.93(s,3H),0.91(s,3H),0.89(s,3H),0.77(s ,3H),0.72(s,3H). ESI-MS: m/z 552.4[MH] - .
实施例148Example 148
N-(2-(N,N-二羟乙基)氨基乙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-76)N-(2-(N,N-Dihydroxyethyl)aminoethyl)-3β-hydroxy-oleanane-13(18)-ene-28-amide (compound A-76)
Figure PCTCN2020093248-appb-000186
Figure PCTCN2020093248-appb-000186
参照实施例13的方法，将3-二甲氨基丙胺替换成N,N-双(2-羟乙基)乙二胺，制得化合物A-76： 1H NMR(300MHz,DMSO-d 6)δ6.61–6.53(m,1H),4.42–4.32(m,2H),4.30(d,J＝5.0Hz,1H),3.49–3.38(m,4H),3.27–3.16(m,1H),3.15–2.96(m,2H),2.85–2.72(m,1H),2.61–2.54(m,4H),2.43(d,J＝14.1Hz,1H),2.25–2.15(m,1H),1.15(s,3H),0.91(s,3H),0.90(s,6H),0.85(s,3H),0.72(s,3H),0.69(s,3H).ESI-MS:m/z 587.5[M+H] +。 Referring to the method of Example 13, the 3-dimethylaminopropylamine was replaced with N,N-bis(2-hydroxyethyl)ethylenediamine to obtain compound A-76: 1 H NMR (300MHz, DMSO-d 6 ) δ6.61–6.53(m,1H), 4.42–4.32(m,2H), 4.30(d,J=5.0Hz,1H), 3.49–3.38(m,4H), 3.27–3.16(m,1H), 3.15–2.96(m,2H), 2.85–2.72(m,1H), 2.61–2.54(m,4H), 2.43(d,J=14.1Hz,1H), 2.25–2.15(m,1H), 1.15( s,3H),0.91(s,3H),0.90(s,6H),0.85(s,3H),0.72(s,3H),0.69(s,3H).ESI-MS:m/z 587.5(M +H] + .
实施例149Example 149
N-(2-(N,N-二羟乙基)氨基乙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺(化合物B-15)N-(2-(N,N-Dihydroxyethyl)aminoethyl)-3β-hydroxy-oleanane-12-ene-28-amide (compound B-15)
Figure PCTCN2020093248-appb-000187
Figure PCTCN2020093248-appb-000187
参照实施例85的方法，将4-(2-氨乙基)硫代吗啉-1,1-二氧化物替换成N,N-双(2-羟乙基)乙二胺，制得化合物B-15： 1H NMR(300MHz,DMSO-d 6)δ7.14–7.05(m,1H),5.27–5.22(m,1H),4.45–4.33(m,2H),4.29(d,J＝5.1Hz,1H),3.49 –3.39(m,4H),3.23–3.11(m,1H),3.07–2.92(m,2H),2.80–2.69(m,1H),2.63–2.54(m,4H),1.11(s,3H),0.91(s,3H),0.90(s,6H),0.87(s,3H),0.69(s,6H).ESI-MS:m/z 587.5[M+H] +。 Referring to the method of Example 85, 4-(2-aminoethyl)thiomorpholine-1,1-dioxide was replaced with N,N-bis(2-hydroxyethyl)ethylenediamine to obtain the compound B-15: 1 H NMR (300MHz, DMSO-d 6 ) δ7.14–7.05(m,1H), 5.27–5.22(m,1H), 4.45–4.33(m,2H), 4.29(d,J= 5.1Hz, 1H), 3.49 -3.39 (m, 4H), 3.23-3.11 (m, 1H), 3.07-2.92 (m, 2H), 2.80-2.69 (m, 1H), 2.63-2.54 (m, 4H) ,1.11(s,3H),0.91(s,3H),0.90(s,6H),0.87(s,3H),0.69(s,6H).ESI-MS:m/z 587.5[M+H] + .
实施例150Example 150
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-乙酰氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-108)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-acetoxy-oleanorane-13(18)-ene-28-amide (compound A-108)
Figure PCTCN2020093248-appb-000188
Figure PCTCN2020093248-appb-000188
参照实施例2的方法，制得化合物II-1。取化合物II-1(150mg，0.300mmol)溶于四氢呋喃(8mL)中，依次加入N,N-二异丙基乙胺(50μL，0.300mmol)和O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸(116mg，0.361mmol)，室温搅拌反应。TLC检测反应完全后，减压蒸除溶剂，残余物用乙酸乙酯(8mL)溶解，水洗(8mL×2)，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝20：1)纯化，得化合物XIX-1，直接用于下一步。According to the method of Example 2, compound II-1 was prepared. Dissolve compound II-1 (150mg, 0.300mmol) in tetrahydrofuran (8mL), add N,N-diisopropylethylamine (50μL, 0.300mmol) and O-benzotriazole-N,N, N',N'-tetramethylurea tetrafluoroboric acid (116mg, 0.361mmol), the reaction was stirred at room temperature. After the completion of the reaction was detected by TLC, the solvent was evaporated under reduced pressure. The residue was dissolved in ethyl acetate (8mL), washed with water (8mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to silica gel column chromatography (petroleum ether: Ethyl acetate = 20:1) Purify to obtain compound XIX-1, which is directly used in the next step.
将化合物XIX-1溶于N,N-二甲基甲酰胺(8mL)中，依次加入4-(3-氨丙基)硫代吗啉-1,1-二氧化物(58mg，0.300mmol)和碳酸钠(32mg，0.300mmol)，室温搅拌反应。TLC检测反应完全后，用乙酸乙酯(8mL)稀释反应液，水洗(8mL)反应液，水层用乙酸乙酯(8mL)萃取，合并有机相，水(8mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物A-108(白色固体，179mg，产率89％)： 1H NMR(300MHz,CDCl 3)δ5.99(t,J＝5.8Hz,1H),4.51(dd,J＝10.6,5.7Hz,1H),3.34–3.22(m,2H),3.09–2.93(m,8H),2.87–2.76(m,1H),2.57–2.34(m,4H),2.05(s,3H),1.19(s,3H),0.90(s,9H),0.87(s,3H),0.85(s,3H),0.74(s,3H).ESI-MS:m/z 671.5[M-H] -。 Compound XIX-1 was dissolved in N,N-dimethylformamide (8mL), and 4-(3-aminopropyl)thiomorpholine-1,1-dioxide (58mg, 0.300mmol) was added successively And sodium carbonate (32mg, 0.300mmol), and the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, the reaction solution was diluted with ethyl acetate (8mL), washed with water (8mL), the aqueous layer was extracted with ethyl acetate (8mL), the organic phases were combined, washed with water (8mL×2), and anhydrous sulfuric acid The sodium was dried, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound A-108 (white solid, 179 mg, yield 89%): 1 H NMR (300MHz, CDCl 3 )δ5.99(t,J=5.8Hz,1H), 4.51(dd,J=10.6,5.7Hz,1H), 3.34–3.22(m,2H), 3.09–2.93(m,8H), 2.87– 2.76(m,1H), 2.57--2.34(m,4H), 2.05(s,3H), 1.19(s,3H), 0.90(s,9H), 0.87(s,3H), 0.85(s,3H) ,0.74(s,3H). ESI-MS: m/z 671.5[MH] - .
实施例151Example 151
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-(3-羧基丙酰)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-109)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-(3-carboxypropionyl)oxy-oleanorane-13(18)-en-28-amide (Compound A-109)
Figure PCTCN2020093248-appb-000189
Figure PCTCN2020093248-appb-000189
参照实施例13的方法，将3-二甲氨基丙胺替换成4-(3-氨丙基)硫代吗啉-1,1-二氧化物，邻苯二甲酸酐替换成丁二酸酐，制得化合物A-109： 1H NMR(300MHz,CDCl 3)δ6.06–5.95(m,1H),4.59–4.48(m,1H),3.39–3.18(m,2H),3.13–2.91(m,8H),2.87–2.75(m,1H),2.73–2.58(m,4H),2.59–2.33(m,4H),1.19(s,3H),0.90(s,9H),0.87(s,3H),0.85(s,3H),0.74(s,3H).ESI-MS:m/z 729.5[M-H] -。 Refer to the method of Example 13, replace 3-dimethylaminopropylamine with 4-(3-aminopropyl)thiomorpholine-1,1-dioxide, and replace phthalic anhydride with succinic anhydride to prepare Obtain compound A-109: 1 H NMR (300MHz, CDCl 3 ) δ 6.06–5.95 (m, 1H), 4.59–4.48 (m, 1H), 3.39–3.18 (m, 2H), 3.13–2.91 (m, 8H), 2.87–2.75(m,1H), 2.73–2.58(m,4H), 2.59–2.33(m,4H), 1.19(s,3H), 0.90(s,9H), 0.87(s,3H) , 0.85(s, 3H), 0.74(s, 3H). ESI-MS: m/z 729.5[MH] - .
实施例152Example 152
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-(3-吡啶甲酰)氧基-齐墩果烷-13(18)-烯-28-酰胺(化合物A-110)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-(3-pyridinecarbonyl)oxy-oleanorane-13(18)-en-28-amide (Compound A-110)
Figure PCTCN2020093248-appb-000190
Figure PCTCN2020093248-appb-000190
取化合物A-81(100mg，0.158mmol)溶于无水二氯甲烷(8mL)中，依次加入N,N'-二环己基碳酰亚胺(66mg，0.317mmol)、4-二甲氨基吡啶(20mg，0.158mmol)和烟酸(30mg，0.238mmol)，室温搅拌反应。TLC检测反应完全后，抽滤，滤液用水(8mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物A-110(白色固体，83mg，产率72％)： 1H NMR(300MHz,methanol-d 4)δ5.99(t,J＝5.8Hz,1H),4.51(dd,J＝10.6,5.7Hz,1H),3.34–3.22(m,2H),3.09–2.93(m,8H),2.87–2.76(m,1H),2.57–2.34(m,4H),2.05(s,3H),1.19(s,3H),0.90(s,9H),0.87(s,3H),0.85(s,3H),0.74(s,3H).ESI-MS:m/z 736.6[M+H] +。 Dissolve compound A-81 (100mg, 0.158mmol) in anhydrous dichloromethane (8mL), add N,N'-dicyclohexylcarbimide (66mg, 0.317mmol), 4-dimethylaminopyridine in turn (20mg, 0.158mmol) and niacin (30mg, 0.238mmol), the reaction was stirred at room temperature. After the completion of the reaction was detected by TLC, it was suction filtered, the filtrate was washed with water (8mL×2), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound A -110 (white solid, 83mg, yield 72%): 1 H NMR (300MHz, methanol-d 4 ) δ 5.99 (t, J = 5.8 Hz, 1H), 4.51 (dd, J = 10.6, 5.7 Hz, 1H), 3.34--3.22(m, 2H), 3.09--2.93(m, 8H), 2.87--2.76(m, 1H), 2.57--2.34(m, 4H), 2.05(s, 3H), 1.19(s, 3H), 0.90 (s, 9H), 0.87 (s, 3H), 0.85 (s, 3H), 0.74 (s, 3H). ESI-MS: m/z 736.6 [M+H] + .
实施例153Example 153
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-(2S-氨基-3-甲基丁酰)氧基-齐墩果烷-13(18)-烯-28-酰胺二盐酸盐(化合物A-111)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-(2S-amino-3-methylbutyryl)oxy-oleanorane-13(18)- En-28-amide dihydrochloride (Compound A-111)
Figure PCTCN2020093248-appb-000191
Figure PCTCN2020093248-appb-000191
Figure PCTCN2020093248-appb-000192
Figure PCTCN2020093248-appb-000192
取化合物A-81(100mg，0.158mmol)溶于无水二氯甲烷(8mL)中，依次加入N,N'-二环己基碳酰亚胺(66mg，0.317mmol)、4-二甲氨基吡啶(20mg，0.158mmol)和N-Boc-L-缬氨酸(52mg，0.238mmol)，室温搅拌反应。TLC检测反应完全后，抽滤，滤液用水(8mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物XX-1(白色固体，127mg，产率97％)。Dissolve compound A-81 (100mg, 0.158mmol) in anhydrous dichloromethane (8mL), add N,N'-dicyclohexylcarbimide (66mg, 0.317mmol), 4-dimethylaminopyridine in turn (20mg, 0.158mmol) and N-Boc-L-valine (52mg, 0.238mmol), the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, suction filtration, the filtrate was washed with water (8mL×2), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound XX -1 (white solid, 127 mg, yield 97%).
将化合物XX-1(127mg，0.158mmol)溶于无水乙醇(10mL)中，通入氯化氢气体30分钟，室温搅拌反应。TLC检测反应完全后，减压蒸除溶剂，残余物用乙酸乙酯和甲醇重结晶，得化合物A-111(白色固体，95mg，产率77％)： 1H NMR(300MHz,methanol-d 4)δ4.77–4.69(m,1H),4.00(d,J＝3.8Hz,1H),3.34–3.13(m,8H),2.95(d,J＝13.9Hz,1H),2.87–2.71(m,2H),2.61(d,J＝14.3Hz,1H),2.46–2.37(m,1H),2.37–2.27(m,1H),1.29(s,3H),1.06(s,3H),1.04(s,3H),1.01(s,3H),0.99(s,6H),0.83(s,3H).ESI-MS:m/z 730.6[M+H] +。 Compound XX-1 (127 mg, 0.158 mmol) was dissolved in absolute ethanol (10 mL), hydrogen chloride gas was bubbled in for 30 minutes, and the reaction was stirred at room temperature. After the completion of the reaction was detected by TLC, the solvent was evaporated under reduced pressure, and the residue was recrystallized from ethyl acetate and methanol to obtain compound A-111 (white solid, 95 mg, yield 77%): 1 H NMR (300MHz, methanol-d 4 )δ4.77–4.69(m,1H), 4.00(d,J=3.8Hz,1H), 3.34–3.13(m,8H), 2.95(d,J=13.9Hz,1H), 2.87–2.71(m ,2H), 2.61(d,J=14.3Hz,1H), 2.46–2.37(m,1H), 2.37–2.27(m,1H),1.29(s,3H),1.06(s,3H),1.04( s, 3H), 1.01 (s, 3H), 0.99 (s, 6H), 0.83 (s, 3H). ESI-MS: m/z 730.6 [M+H] + .
实施例154Example 154
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-羟基-齐墩果烷-13(18)-烯-28-酰胺盐酸盐(化合物A-112)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-hydroxy-oleanorane-13(18)-ene-28-amide hydrochloride (Compound A-112 )
Figure PCTCN2020093248-appb-000193
Figure PCTCN2020093248-appb-000193
将化合物A-81(100mg，0.158mmol)溶于无水乙醇(10mL)中，通入氯化氢气体30分钟，室温搅拌反应。反应完全后减压蒸除溶剂直至剩余2mL，抽滤，滤饼用乙酸乙酯(1mL×2)洗涤，烘干，得化合物A-111(白色固体，52mg，产率49％) 1H NMR(300MHz,DMSO-d 6)δ6.99–6.91(m,1H),3.90–3.51(m,8H),3.21–3.08(m,4H),3.06–2.97(m,1H),2.77(d,J＝13.7Hz,1H),2.42(d,J＝13.7Hz,1H),2.21–2.11(m,1H),1.13(s,3H),0.89(s,9H),0.83(s,3H),0.71(s,3H),0.67(s,3H).ESI-MS:m/z 631.5[M+H] +。 Compound A-81 (100 mg, 0.158 mmol) was dissolved in absolute ethanol (10 mL), hydrogen chloride gas was bubbled in for 30 minutes, and the reaction was stirred at room temperature. After the reaction was completed, the solvent was evaporated under reduced pressure until 2 mL remained, and filtered with suction. The filter cake was washed with ethyl acetate (1 mL×2) and dried to obtain compound A-111 (white solid, 52 mg, yield 49%) 1 H NMR (300MHz, DMSO-d 6 )δ 6.99-6.91(m, 1H), 3.90-3.51(m, 8H), 3.21-3.08(m, 4H), 3.06-2.97(m, 1H), 2.77(d, J = 13.7Hz, 1H), 2.42 (d, J = 13.7Hz, 1H), 2.21–2.11 (m, 1H), 1.13 (s, 3H), 0.89 (s, 9H), 0.83 (s, 3H), 0.71 (s, 3H), 0.67 (s, 3H). ESI-MS: m/z 631.5 [M+H] + .
实施例155Example 155
3β-羟基-齐墩果烷-13(18)-烯-28-酸3-(1,1-二氧代硫代吗啉基)丙酯(化合物A-113)3β-Hydroxy-oleanorane-13(18)-ene-28-acid 3-(1,1-dioxothiomorpholinyl)propyl ester (Compound A-113)
Figure PCTCN2020093248-appb-000194
Figure PCTCN2020093248-appb-000194
取化合物II-1(200mg，0.401mmol)溶于N,N-二甲基甲酰胺(8mL)中，依次加入3-溴-1-丙醇(73μL，0.802mmol)和碳酸钾(111mg，0.802mmol)，室温搅拌反应。TLC检测反应完全后，用乙酸乙酯(8mL)稀释反应液，水洗(8mL)反应液，水层用乙酸乙酯(8mL)萃取，合并有机相，水(8mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝5：1)纯化，得化合物XXI-1(白色固体，197mg，产率88％)。Dissolve compound II-1 (200mg, 0.401mmol) in N,N-dimethylformamide (8mL), add 3-bromo-1-propanol (73μL, 0.802mmol) and potassium carbonate (111mg, 0.802) in sequence mmol), the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, the reaction solution was diluted with ethyl acetate (8mL), washed with water (8mL), the aqueous layer was extracted with ethyl acetate (8mL), the organic phases were combined, washed with water (8mL×2), and anhydrous sulfuric acid Dry with sodium, filter, concentrate the filtrate, and purify by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound XXI-1 (white solid, 197 mg, yield 88%).
取化合物XXI-1(197mg，0.354mmol)溶于二氯甲烷(8mL)中，加入氯铬酸吡啶盐(153mg，0.707mmol)，室温搅拌反应。TLC检测反应完全后，过滤硅藻土，滤液旋干，经硅胶柱层析(石油醚：乙酸乙酯＝10：1)纯化，得化合物XXI-2(白色固体，157mg，产率80％)。Compound XXI-1 (197 mg, 0.354 mmol) was dissolved in dichloromethane (8 mL), pyridinium chlorochromate (153 mg, 0.707 mmol) was added, and the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, the celite was filtered, the filtrate was spin-dried, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound XXI-2 (white solid, 157 mg, yield 80%) .
将化合物XXI-2(157mg，0.283mmol)和1,1-二氧化硫代吗啉(39mg，0.283mmol)溶于二氯甲烷(8mL)中，室温搅拌1小时后加入三乙酰氧基硼氢化钠(90mg，0.424mmol)，室温搅拌反应。TLC检测反应完全后，加入饱和碳酸氢钠溶液(5mL)，分液，水(5mL×2)洗有机相，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝3：1)纯化，得化合物XXI-3(白色固体，69mg，产率36％)。Compound XXI-2 (157mg, 0.283mmol) and 1,1-thiomorpholine dioxide (39mg, 0.283mmol) were dissolved in dichloromethane (8mL), stirred at room temperature for 1 hour, and sodium triacetoxyborohydride ( 90mg, 0.424mmol), the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, saturated sodium bicarbonate solution (5mL) was added, the layers were separated, the organic phase was washed with water (5mL×2), dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to silica gel column chromatography (petroleum ether: acetic acid) Ethyl ester = 3: 1) Purification to obtain compound XXI-3 (white solid, 69 mg, yield 36%).
取化合物XXI-3(69mg，0.102mmol)溶于甲醇(5mL)和四氢呋喃(5mL)的混合溶液中，加入氢氧化钾(57mg，1.024mmol)，室温搅拌反应。TLC检测反应完全后，减压蒸除溶剂，残余物用乙酸乙酯(10mL)溶解，水(10mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝ 3：1)纯化，得化合物A-113(白色固体，45mg，产率69％)： 1H NMR(300MHz,CDCl 3)δ4.21–4.02(m,2H),3.28–3.17(m,1H),3.14–2.89(m,8H),2.74(d,J＝11.3Hz,1H),2.57(t,J＝7.0Hz,2H),2.43(d,J＝14.0Hz,1H),2.22–2.11(m,1H),1.16(s,3H),0.99(s,3H),0.90(s,3H),0.90(s,3H),0.87(s,3H),0.77(s,3H),0.74(s,3H).ESI-MS:m/z 632.5[M+H] +。 Compound XXI-3 (69 mg, 0.102 mmol) was dissolved in a mixed solution of methanol (5 mL) and tetrahydrofuran (5 mL), potassium hydroxide (57 mg, 1.024 mmol) was added, and the reaction was stirred at room temperature. After the completion of the reaction was detected by TLC, the solvent was evaporated under reduced pressure. The residue was dissolved in ethyl acetate (10 mL), washed with water (10 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to silica gel column chromatography (petroleum ether). : Ethyl acetate = 3: 1) Purification to obtain compound A-113 (white solid, 45 mg, yield 69%): 1 H NMR (300MHz, CDCl 3 ) δ 4.21–4.02 (m, 2H), 3.28– 3.17(m,1H),3.14–2.89(m,8H),2.74(d,J=11.3Hz,1H), 2.57(t,J=7.0Hz,2H),2.43(d,J=14.0Hz,1H ), 2.22--2.11(m, 1H), 1.16(s, 3H), 0.99(s, 3H), 0.90(s, 3H), 0.90(s, 3H), 0.87(s, 3H), 0.77(s, 3H), 0.74(s, 3H). ESI-MS: m/z 632.5 [M+H] + .
实施例156Example 156
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-乙酰氧基-齐墩果烷-12-烯-28-酰胺(化合物B-121)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-acetoxy-oleanorane-12-en-28-amide (compound B-121)
Figure PCTCN2020093248-appb-000195
Figure PCTCN2020093248-appb-000195
参照实施例73的方法，制得化合物XI-1。取化合物XI-1(150mg，0.300mmol)溶于四氢呋喃(8mL)中，依次加入N,N-二异丙基乙胺(50μL，0.300mmol)和O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸(116mg，0.361mmol)，室温搅拌反应。TLC检测反应完全后，减压蒸除溶剂，残余物用乙酸乙酯(8mL)溶解，水洗(8mL×2)，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝20：1)纯化，得化合物XXII-1，直接用于下一步。Referring to the method of Example 73, compound XI-1 was prepared. Dissolve compound XI-1 (150mg, 0.300mmol) in tetrahydrofuran (8mL), add N,N-diisopropylethylamine (50μL, 0.300mmol) and O-benzotriazole-N,N, N',N'-tetramethylurea tetrafluoroboric acid (116mg, 0.361mmol), the reaction was stirred at room temperature. After the completion of the reaction was detected by TLC, the solvent was evaporated under reduced pressure. The residue was dissolved in ethyl acetate (8mL), washed with water (8mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to silica gel column chromatography (petroleum ether: Ethyl acetate = 20:1) Purify to obtain compound XXII-1, which is directly used in the next step.
将化合物XXII-1溶于N,N-二甲基甲酰胺(8mL)中，依次加入4-(3-氨丙基)硫代吗啉-1,1-二氧化物(58mg，0.300mmol)和碳酸钠(32mg，0.300mmol)，室温搅拌反应。TLC检测反应完全后，用乙酸乙酯(8mL)稀释反应液，水洗(8mL)反应液，水层用乙酸乙酯(8mL)萃取，合并有机相，水(8mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物B-121(白色固体，172mg，产率85％)： 1H NMR(300MHz,CDCl 3)δ6.10–5.97(m,1H),5.41–5.33(m,1H),4.54–4.43(m,1H),3.50–3.34(m,1H),3.17–2.92(m,8H),2.63–2.43(m,3H),2.05(s,3H),1.16(s,3H),0.93(s,3H),0.91(s,3H),0.90(s,3H),0.87(s,3H),0.86(s,3H),0.76(s,3H).ESI-MS:m/z673.6[M+H] +。 Compound XXII-1 was dissolved in N,N-dimethylformamide (8mL), and 4-(3-aminopropyl)thiomorpholine-1,1-dioxide (58mg, 0.300mmol) was added sequentially And sodium carbonate (32mg, 0.300mmol), and the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, the reaction solution was diluted with ethyl acetate (8mL), washed with water (8mL), the aqueous layer was extracted with ethyl acetate (8mL), the organic phases were combined, washed with water (8mL×2), and anhydrous sulfuric acid Dry with sodium, filter, concentrate the filtrate, and purify by silica gel column chromatography (dichloromethane: methanol=20:1) to obtain compound B-121 (white solid, 172mg, yield 85%): 1 H NMR (300MHz, CDCl 3 )δ6.10-5.97(m,1H), 5.41-5.33(m,1H), 4.54-4.43(m,1H), 3.50-3.34(m,1H), 3.17-2.92(m,8H), 2.63 --2.43(m,3H),2.05(s,3H),1.16(s,3H),0.93(s,3H),0.91(s,3H),0.90(s,3H),0.87(s,3H), 0.86 (s, 3H), 0.76 (s, 3H). ESI-MS: m/z 673.6 [M+H] + .
实施例157Example 157
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-(3-羧基丙酰)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-122)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-(3-carboxypropionyl)oxy-oleanorane-12-en-28-amide (compound B -122)
Figure PCTCN2020093248-appb-000196
Figure PCTCN2020093248-appb-000196
将化合物B-20(100mg，0.158mmol)溶于无水吡啶(8mL)中，加入丁二酸酐(159mg，1.585mmol)和4-二甲氨基吡啶(20mg，0.158mmol)，反应液升温至115℃搅拌过夜。TLC检测反应完全后，加入二氯甲烷(20mL)稀释反应液，反应液依次用1N稀盐酸(10mL×3)和水(10mL×3)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物B-122(白色固体，64mg，产率55％)： 1H NMR(300MHz,CDCl 3)δ6.07–5.97(m,1H),5.39–5.33(m,1H),4.59–4.45(m,1H),3.50–3.34(m,1H),3.12–2.90(m,8H),2.74–2.57(m,4H),2.57–2.42(m,4H),1.16(s,3H),0.93(s,3H),0.91(s,3H),0.90(s,3H),0.86(s,6H),0.76(s,3H).ESI-MS:m/z 731.6[M+H] +。 Compound B-20 (100 mg, 0.158 mmol) was dissolved in anhydrous pyridine (8 mL), succinic anhydride (159 mg, 1.585 mmol) and 4-dimethylaminopyridine (20 mg, 0.158 mmol) were added, and the reaction solution was heated to 115 Stir overnight at °C. After the completion of the reaction detected by TLC, dichloromethane (20mL) was added to dilute the reaction solution. The reaction solution was washed with 1N dilute hydrochloric acid (10mL×3) and water (10mL×3) successively, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound B-122 (white solid, 64mg, yield 55%): 1 H NMR (300MHz, CDCl 3 ) δ 6.07–5.97 (m ,1H),5.39–5.33(m,1H),4.59–4.45(m,1H), 3.50–3.34(m,1H), 3.12–2.90(m,8H), 2.74–2.57(m,4H), 2.57 --2.42(m,4H),1.16(s,3H),0.93(s,3H),0.91(s,3H),0.90(s,3H),0.86(s,6H),0.76(s,3H). ESI-MS: m/z 731.6 [M+H] + .
实施例158Example 158
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-(3-吡啶甲酰)氧基-齐墩果烷-12-烯-28-酰胺(化合物B-123)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-(3-pyridinecarbonyl)oxy-oleanorane-12-en-28-amide (compound B -123)
Figure PCTCN2020093248-appb-000197
Figure PCTCN2020093248-appb-000197
取化合物B-20(141mg，0.223mmol)溶于无水二氯甲烷(8mL)中，依次加入N,N'-二环己基碳酰亚胺(184mg，0.894mmol)、4-二甲氨基吡啶(28mg，0.223mmol)和烟酸(55mg，0.447mmol)，室温搅拌反应。TLC检测反应完全后，抽滤，滤液用水(8mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物B-123(白色固体，118mg，产率72％)： 1H NMR(300MHz,CDCl 3)δ9.23(s,1H),8.77(d,J＝4.7Hz,1H),8.29(d,J＝8.0Hz,1H),7.39(dd,J＝8.0,4.9Hz,1H),6.07–5.96(m,1H),5.41 –5.33(m,1H),4.83–4.70(m,1H),3.51–3.34(m,1H),3.11–2.92(m,8H),2.59–2.45(m,3H),1.19(s,3H),1.02(s,3H),0.99(s,3H),0.95(s,3H),0.92(s,3H),0.91(s,3H),0.78(s,3H).ESI-MS:m/z 736.6[M+H] +。 Dissolve compound B-20 (141mg, 0.223mmol) in anhydrous dichloromethane (8mL), add N,N'-dicyclohexylcarbimide (184mg, 0.894mmol) and 4-dimethylaminopyridine in sequence (28mg, 0.223mmol) and niacin (55mg, 0.447mmol), the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, it was suction filtered, the filtrate was washed with water (8mL×2), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound B -123 (white solid, 118mg, yield 72%): 1 H NMR (300MHz, CDCl 3 ) δ 9.23 (s, 1H), 8.77 (d, J = 4.7 Hz, 1H), 8.29 (d, J = 8.0Hz,1H),7.39(dd,J=8.0,4.9Hz,1H),6.07–5.96(m,1H),5.41 –5.33(m,1H),4.83–4.70(m,1H),3.51–3.34 (m,1H),3.11-2.92(m,8H),2.59-2.45(m,3H),1.19(s,3H),1.02(s,3H),0.99(s,3H),0.95(s,3H) ), 0.92 (s, 3H), 0.91 (s, 3H), 0.78 (s, 3H). ESI-MS: m/z 736.6 [M+H] + .
实施例159Example 159
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-(2S-氨基-3-甲基丁酰)氧基-齐墩果烷-12-烯-28-酰胺二盐酸盐(化合物B-124)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-(2S-amino-3-methylbutyryl)oxy-oleanorane-12-ene-28 -Amide dihydrochloride (compound B-124)
Figure PCTCN2020093248-appb-000198
Figure PCTCN2020093248-appb-000198
取化合物B-20(100mg，0.158mmol)溶于无水二氯甲烷(8mL)中，依次加入N,N'-二环己基碳酰亚胺(66mg，0.317mmol)、4-二甲氨基吡啶(20mg，0.158mmol)和N-Boc-L-缬氨酸(52mg，0.238mmol)，室温搅拌反应。TLC检测反应完全后，抽滤，滤液用水(8mL×2)洗涤，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(二氯甲烷：甲醇＝20：1)纯化，得化合物XXIII-1(白色固体，125mg，产率95％)。Dissolve compound B-20 (100mg, 0.158mmol) in anhydrous dichloromethane (8mL), add N,N'-dicyclohexylcarbimide (66mg, 0.317mmol) and 4-dimethylaminopyridine in turn (20mg, 0.158mmol) and N-Boc-L-valine (52mg, 0.238mmol), the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, it was suction filtered, the filtrate was washed with water (8mL×2), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 20:1) to obtain compound XXIII -1 (white solid, 125 mg, yield 95%).
将化合物XXIII-1(125mg，0.158mmol)溶于无水乙醇(10mL)中，通入氯化氢气体30分钟，室温搅拌反应。TLC检测反应完全后，减压蒸除溶剂，残余物用乙酸乙酯打浆，抽滤，滤饼用乙酸乙酯(2ml×2)洗涤，烘干，得化合物B-124(白色固体，110mg，产率91％)： 1H NMR(300MHz,methanol-d 4)δ5.45–5.39(m,1H),4.76–4.65(m,1H),4.00(d,J＝3.7Hz,1H),3.64–3.40(m,8H),3.32–3.18(m,1H),3.17–2.94(m,2H),2.93–2.82(m,1H),2.49–2.33(m,1H),1.25(s,3H),1.05(s,3H),1.01(s,6H),1.00(s,3H),0.97(s,3H),0.86(s,3H).ESI-MS:m/z 730.6[M+H] +。 Compound XXIII-1 (125 mg, 0.158 mmol) was dissolved in absolute ethanol (10 mL), hydrogen chloride gas was bubbled in for 30 minutes, and the reaction was stirred at room temperature. After the completion of the reaction was detected by TLC, the solvent was evaporated under reduced pressure, the residue was slurried with ethyl acetate, filtered with suction, the filter cake was washed with ethyl acetate (2ml×2), and dried to obtain compound B-124 (white solid, 110mg, Yield 91%): 1 H NMR (300MHz, methanol-d 4 ) δ 5.45–5.39 (m, 1H), 4.76–4.65 (m, 1H), 4.00 (d, J=3.7 Hz, 1H), 3.64 --3.40(m,8H), 3.32–3.18(m,1H), 3.17–2.94(m,2H), 2.93–2.82(m,1H), 2.49–2.33(m,1H),1.25(s,3H) ,1.05(s,3H),1.01(s,6H),1.00(s,3H),0.97(s,3H),0.86(s,3H).ESI-MS:m/z 730.6[M+H] + .
实施例160Example 160
N-(3-(1,1-二氧代硫代吗啉基)丙基)-3β-羟基-齐墩果烷-12-烯-28-酰胺盐酸盐(化合物B-125)N-(3-(1,1-dioxothiomorpholinyl)propyl)-3β-hydroxy-oleanane-12-en-28-amide hydrochloride (compound B-125)
Figure PCTCN2020093248-appb-000199
Figure PCTCN2020093248-appb-000199
Figure PCTCN2020093248-appb-000200
Figure PCTCN2020093248-appb-000200
将化合物B-20(70mg，0.111mmol)溶于无水乙醇(10mL)中，通入氯化氢气体30分钟，室温搅拌反应。反应完全后减压蒸除溶剂，残余物用乙酸乙酯打浆，抽滤，滤饼用乙酸乙酯(2ml×2)洗涤，烘干，得化合物B-125(白色固体，69mg，产率93％)： 1H NMR(300MHz,methanol-d 4)δ5.47–5.41(m,1H),3.96–3.78(m,4H),3.73–3.60(m,4H),3.46–3.26(m,4H),3.26–3.17(m,1H),2.92–2.82(m,1H),1.25(s,3H),1.04(s,3H),1.03(s,3H),1.01(s,3H),0.99(s,3H),0.85(s,3H),0.84(s,3H).ESI-MS:m/z 631.6[M+H] +。 Compound B-20 (70 mg, 0.111 mmol) was dissolved in absolute ethanol (10 mL), hydrogen chloride gas was bubbled in for 30 minutes, and the reaction was stirred at room temperature. After the reaction was complete, the solvent was evaporated under reduced pressure, the residue was slurried with ethyl acetate, filtered with suction, the filter cake was washed with ethyl acetate (2ml×2), and dried to obtain compound B-125 (white solid, 69mg, yield 93 %): 1 H NMR(300MHz, methanol-d 4 )δ5.47–5.41(m,1H), 3.96–3.78(m,4H), 3.73–3.60(m,4H), 3.46–3.26(m,4H) ), 3.26–3.17(m,1H), 2.92–2.82(m,1H),1.25(s,3H),1.04(s,3H),1.03(s,3H),1.01(s,3H),0.99( s, 3H), 0.85 (s, 3H), 0.84 (s, 3H). ESI-MS: m/z 631.6 [M+H] + .
实施例161Example 161
3β-羟基-齐墩果烷-12-烯-28-酸3-(1,1-二氧代硫代吗啉基)丙酯盐酸盐(化合物B-126)3β-Hydroxy-oleanan-12-ene-28-acid 3-(1,1-dioxothiomorpholinyl) propyl ester hydrochloride (compound B-126)
Figure PCTCN2020093248-appb-000201
Figure PCTCN2020093248-appb-000201
取化合物XI-1(200mg，0.401mmol)溶于N,N-二甲基甲酰胺(8mL)中，依次加入3-溴-1-丙醇(73μL，0.802mmol)和碳酸钾(111mg，0.802mmol)，室温搅拌反应。TLC检测反应完全后，用乙酸乙酯(8mL)稀释反应液，水洗(8mL)反应液，水层用乙酸乙酯(8mL)萃取，合并有机相，水(8mL×2) 洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝5：1)纯化，得化合物XXIV-1(白色固体，220mg，产率98％)。Take compound XI-1 (200mg, 0.401mmol) and dissolve it in N,N-dimethylformamide (8mL), add 3-bromo-1-propanol (73μL, 0.802mmol) and potassium carbonate (111mg, 0.802) in sequence mmol), the reaction was stirred at room temperature. After the completion of the reaction was detected by TLC, the reaction solution was diluted with ethyl acetate (8mL) and washed with water (8mL). The aqueous layer was extracted with ethyl acetate (8mL). The organic phases were combined and washed with water (8mL×2). Anhydrous sulfuric acid Dry with sodium, filter, concentrate the filtrate, and purify by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound XXIV-1 (white solid, 220 mg, yield 98%).
取化合物XXIV-1(220mg，0.395mmol)溶于二氯甲烷(10mL)中，加入氯铬酸吡啶盐(170mg，0.790mmol)，室温搅拌反应。TLC检测反应完全后，过滤硅藻土，滤液旋干，经硅胶柱层析(石油醚：乙酸乙酯＝4：1)纯化，得化合物XXIV-2(白色固体，164mg，产率75％)。The compound XXIV-1 (220 mg, 0.395 mmol) was dissolved in dichloromethane (10 mL), pyridinium chlorochromate (170 mg, 0.790 mmol) was added, and the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, the diatomaceous earth was filtered, the filtrate was spin-dried, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1) to obtain compound XXIV-2 (white solid, 164 mg, yield 75%) .
将化合物XXIV-2(220mg，0.396mmol)和1,1-二氧化硫代吗啉(54mg，0.396mmol)溶于二氯甲烷(8mL)中，室温搅拌1小时后加入三乙酰氧基硼氢化钠(126mg，0.595mmol)，室温搅拌反应。TLC检测反应完全后，加入饱和碳酸氢钠溶液(5mL)，分液，水(5mL×2)洗有机相，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝3：1)纯化，得化合物XXIV-3(白色固体，131mg，产率49％)。Compound XXIV-2 (220mg, 0.396mmol) and 1,1-thiomorpholine dioxide (54mg, 0.396mmol) were dissolved in dichloromethane (8mL), stirred at room temperature for 1 hour, and sodium triacetoxyborohydride ( 126mg, 0.595mmol), the reaction was stirred at room temperature. After the completion of the reaction detected by TLC, saturated sodium bicarbonate solution (5mL) was added, the layers were separated, the organic phase was washed with water (5mL×2), dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to silica gel column chromatography (petroleum ether: acetic acid) Ethyl ester=3:1) Purification to obtain compound XXIV-3 (white solid, 131 mg, yield 49%).
取化合物XXIV-3(131mg，0.194mmol)溶于甲醇(5mL)和四氢呋喃(5mL)的混合溶液中，加入氢氧化钾(109mg，1.943mmol)，室温搅拌反应。TLC检测反应完全后，减压蒸除溶剂，残余物用乙酸乙酯(10mL)溶解，水(10mL×2)洗，无水硫酸钠干燥，过滤，滤液浓缩，经硅胶柱层析(石油醚：乙酸乙酯＝2：1)纯化，得化合物XXIV-4(白色固体，75mg，产率61％)。The compound XXIV-3 (131 mg, 0.194 mmol) was dissolved in a mixed solution of methanol (5 mL) and tetrahydrofuran (5 mL), potassium hydroxide (109 mg, 1.943 mmol) was added, and the reaction was stirred at room temperature. After the completion of the reaction was detected by TLC, the solvent was evaporated under reduced pressure. The residue was dissolved in ethyl acetate (10 mL), washed with water (10 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and subjected to silica gel column chromatography (petroleum ether). : Ethyl acetate=2:1) Purify to obtain compound XXIV-4 (white solid, 75mg, yield 61%).
将化合物XXIV-4(75mg，0.119mmol)溶于无水乙醇(10mL)中，通入氯化氢气体30分钟，室温搅拌反应。反应完全后减压蒸除溶剂，残余物用乙酸乙酯打浆，抽滤，滤饼用乙酸乙酯(2ml×2)洗涤，烘干，得化合物B-126(白色固体，78mg，产率60％)： 1H NMR(300MHz,methanol-d 4)δ5.36–5.27(m,1H),4.16(t,J＝6.3Hz,2H),3.28–3.09(m,8H),2.99–2.89(m,1H),2.82–2.71(m,2H),1.23(s,3H),1.03(s,3H),1.01(s,3H),1.00(s,3H),0.97(s,3H),0.84(s,3H),0.83(s,3H).ESI-MS:m/z 654.5[M+Na] +。 Compound XXIV-4 (75 mg, 0.119 mmol) was dissolved in absolute ethanol (10 mL), hydrogen chloride gas was bubbled in for 30 minutes, and the reaction was stirred at room temperature. After the reaction was complete, the solvent was evaporated under reduced pressure, the residue was slurried with ethyl acetate, filtered with suction, the filter cake was washed with ethyl acetate (2ml×2), and dried to obtain compound B-126 (white solid, 78mg, yield 60 %): 1 H NMR(300MHz, methanol-d 4 )δ5.36–5.27(m,1H), 4.16(t,J=6.3Hz,2H), 3.28–3.09(m,8H), 2.99–2.89( m, 1H), 2.82-2.71 (m, 2H), 1.23 (s, 3H), 1.03 (s, 3H), 1.01 (s, 3H), 1.00 (s, 3H), 0.97 (s, 3H), 0.84 (s, 3H), 0.83 (s, 3H). ESI-MS: m/z 654.5 [M+Na] + .
实施例162Example 162
N-(3β-乙酰氧基-齐墩果烷-13(18)-烯-28-酰基)-硫代吗啉-1,1-二氧化物(化合物A-114)N-(3β-Acetoxy-Olean-13(18)-ene-28-acyl)-thiomorpholine-1,1-dioxide (Compound A-114)
Figure PCTCN2020093248-appb-000202
Figure PCTCN2020093248-appb-000202
参照实施例2的方法，将N,N-二甲基乙二胺替换成1,1-二氧化硫代吗啉，制得化合物A-114： 1H NMR(300MHz,CDCl3)δ4.55–4.46(m,1H),4.36–4.13(m,4H),3.05–2.93(m,4H),2.76(d,J＝13.0Hz,1H),2.48(d,J＝13.5Hz,1H), 2.39(d,J＝12.5Hz,1H),2.05(s,3H),1.18(s,3H),0.92(s,3H),0.90(s,3H),0.87(s,3H),0.86(s,3H),0.85(s,3H),0.79(s,3H).ESI-MS:m/z 616.5[M+H] +。 Referring to the method of Example 2, the N,N-dimethylethylenediamine was replaced with 1,1-thiomorpholine dioxide to obtain compound A-114: 1 H NMR (300MHz, CDCl3) δ4.55–4.46( m,1H), 4.36–4.13(m,4H),3.05–2.93(m,4H), 2.76(d,J=13.0Hz,1H), 2.48(d,J=13.5Hz,1H), 2.39(d ,J=12.5Hz,1H),2.05(s,3H),1.18(s,3H),0.92(s,3H),0.90(s,3H),0.87(s,3H),0.86(s,3H) , 0.85 (s, 3H), 0.79 (s, 3H). ESI-MS: m/z 616.5 [M+H] + .
实施例163Example 163
N-(3β-(3-吡啶甲酰)氧基-齐墩果烷-13(18)-烯-28-酰基)-硫代吗啉-1,1-二氧化物(化合物A-115)N-(3β-(3-picolinoyl)oxy-oleanorane-13(18)-ene-28-acyl)-thiomorpholine-1,1-dioxide (Compound A-115)
Figure PCTCN2020093248-appb-000203
Figure PCTCN2020093248-appb-000203
参照实施例152的方法，将化合物A-81替换成化合物A-85，制得化合物A-115： 1H NMR(300MHz,CDCl 3)δ9.27–9.19(m,1H),8.77(d,J＝3.8Hz,1H),8.29(d,J＝7.8Hz,1H),7.47–7.34(m,1H),4.85–4.72(m,1H),4.39–4.11(m,4H),3.13–2.90(m,4H),2.78(d,J＝13.3Hz,1H),2.49(d,J＝13.1Hz,1H),2.43–2.32(m,1H),1.21(s,3H),1.01(s,3H),0.95(s,6H),0.92(s,3H),0.89(s,3H),0.80(s,3H).ESI-MS:m/z 679.5[M+H] +。 According to the method of Example 152, compound A-81 was replaced with compound A-85 to obtain compound A-115: 1 H NMR (300MHz, CDCl 3 ) δ 9.27–9.19 (m, 1H), 8.77 (d, J=3.8Hz,1H), 8.29(d,J=7.8Hz,1H), 7.47–7.34(m,1H), 4.85–4.72(m,1H), 4.39–4.11(m,4H), 3.13–2.90 (m, 4H), 2.78 (d, J = 13.3 Hz, 1H), 2.49 (d, J = 13.1 Hz, 1H), 2.43-2.32 (m, 1H), 1.21 (s, 3H), 1.01 (s, 3H), 0.95(s, 6H), 0.92(s, 3H), 0.89(s, 3H), 0.80(s, 3H). ESI-MS: m/z 679.5[M+H] + .
实施例164Example 164
N-(3β-(3-羧基丙酰)氧基-齐墩果烷-13(18)-烯-28-酰基)-硫代吗啉-1,1-二氧化物(化合物A-116)N-(3β-(3-carboxypropionyl)oxy-oleanorane-13(18)-ene-28-acyl)-thiomorpholine-1,1-dioxide (compound A-116)
Figure PCTCN2020093248-appb-000204
Figure PCTCN2020093248-appb-000204
参照实施例2的方法，将N,N-二甲基乙二胺替换成1,1-二氧化硫代吗啉，邻苯二甲酸酐替换成丁二酸酐，制得化合物A-116： 1H NMR(300MHz,CDCl 3)δ4.59–4.48(m,1H),4.38–4.12(m,4H),3.06–2.92(m,4H),2.83–2.73(m,1H),2.71–2.57(m,4H),2.48(d,J＝13.2Hz,1H),2.38(d,J＝12.4Hz,1H),1.18(s,3H),0.91(s,3H),0.90(s,3H),0.86(s,6H),0.85(s,3H),0.79(s,3H).ESI-MS:m/z 674.5[M+H] +。 Refer to the method of Example 2, replace N,N-dimethylethylenediamine with 1,1-thiomorpholine dioxide, and replace phthalic anhydride with succinic anhydride to obtain compound A-116: 1 H NMR (300MHz, CDCl 3 ) δ4.59-4.48(m,1H), 4.38-4.12(m,4H), 3.06-2.92(m,4H), 2.83-2.73(m,1H), 2.71-2.57(m, 4H), 2.48 (d, J = 13.2Hz, 1H), 2.38 (d, J = 12.4 Hz, 1H), 1.18 (s, 3H), 0.91 (s, 3H), 0.90 (s, 3H), 0.86 ( s, 6H), 0.85 (s, 3H), 0.79 (s, 3H). ESI-MS: m/z 674.5 [M+H] + .
实施例165Example 165
N-(3β-(2S-氨基-3-甲基丁酰)氧基-齐墩果烷-13(18)-烯-28-酰基)-硫代吗啉-1,1-二氧化物盐酸盐(A-117)N-(3β-(2S-amino-3-methylbutyryl)oxy-oleanorane-13(18)-ene-28-acyl)-thiomorpholine-1,1-dioxide salt Acid salt (A-117)
Figure PCTCN2020093248-appb-000205
Figure PCTCN2020093248-appb-000205
参照实施例153的方法，将化合物A-81替换成化合物A-85，制得化合物A-117： 1H NMR(300MHz,DMSO-d 6)δ8.55–8.28(m,3H),5.61(d,J＝8.1Hz,1H),4.63–4.52(m,1H),4.15–3.93(m,4H),3.88(d,J＝3.8Hz,1H),3.22–3.01(m,4H),2.71(d,J＝11.9Hz,1H),2.38(d,J＝12.9Hz,1H),2.31–2.14(m,2H),1.14(s,3H),0.89(s,3H),0.88(s,6H),0.85(s,3H),0.83(s,3H),0.75(s,3H).ESI-MS:m/z 673.6[M+H] +。 According to the method of Example 153, compound A-81 was replaced with compound A-85 to obtain compound A-117: 1 H NMR (300MHz, DMSO-d 6 )δ8.55-8.28(m,3H), 5.61( d,J=8.1Hz,1H), 4.63–4.52(m,1H), 4.15–3.93(m,4H), 3.88(d,J=3.8Hz,1H), 3.22–3.01(m,4H), 2.71 (d,J=11.9Hz,1H),2.38(d,J=12.9Hz,1H),2.31–2.14(m,2H),1.14(s,3H),0.89(s,3H),0.88(s, 6H), 0.85 (s, 3H), 0.83 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 673.6 [M+H] + .
实施例166Example 166
N-(3β-乙酰氧基-齐墩果烷-12-烯-28-酰基)-硫代吗啉-1,1-二氧化物(化合物B-127)N-(3β-acetoxy-oleanorane-12-ene-28-acyl)-thiomorpholine-1,1-dioxide (compound B-127)
Figure PCTCN2020093248-appb-000206
Figure PCTCN2020093248-appb-000206
参照实施例73的方法，将N,N-二甲基乙二胺替换成1,1-二氧化硫代吗啉，制得化合物B-127： 1H NMR(300MHz,CDCl 3)δ5.32–5.25(m,1H),4.53–4.44(m,1H),4.16–4.05(m,4H),3.12–2.90(m,5H),2.21(t,J＝13.6Hz,1H),2.04(s,3H),1.14(s,3H),0.93(s,6H),0.91(s,3H),0.86(s,3H),0.85(s,3H),0.71(s,3H).ESI-MS:m/z 616.5[M+H] +。 Referring to the method of Example 73, N,N-dimethylethylenediamine was replaced with 1,1-thiomorpholine dioxide to obtain compound B-127: 1 H NMR (300MHz, CDCl 3 ) δ5.32–5.25 (m,1H),4.53–4.44(m,1H),4.16–4.05(m,4H),3.12–2.90(m,5H),2.21(t,J=13.6Hz,1H),2.04(s,3H) ), 1.14(s, 3H), 0.93(s, 6H), 0.91(s, 3H), 0.86(s, 3H), 0.85(s, 3H), 0.71(s, 3H).ESI-MS: m/ z 616.5[M+H] + .
实施例167Example 167
N-(3β-(3-吡啶甲酰)氧基-齐墩果烷-12-烯-28-酰基)-硫代吗啉-1,1-二氧化物(化合物B-128)N-(3β-(3-picolinoyl)oxy-oleanorane-12-ene-28-acyl)-thiomorpholine-1,1-dioxide (compound B-128)
Figure PCTCN2020093248-appb-000207
Figure PCTCN2020093248-appb-000207
参照实施例152的方法，将化合物A-81替换成化合物B-9，制得化合物B-128： 1H NMR(300MHz,CDCl 3)δ9.23(s,1H),8.77(d,J＝4.0Hz,1H),8.29(d,J＝8.0Hz,1H),7.42–7.35(m,1H),5.32–5.27(m,1H),4.82–4.72(m,1H),4.18–4.02(m,4H),3.14–2.91(m,5H),2.30–2.13(m,1H),1.17(s,3H),1.02(s,3H),0.98(s,3H),0.94(s,6H),0.92(s,3H),0.74(s,3H).ESI-MS:m/z 679.5[M+H] +。 According to the method of Example 152, compound A-81 was replaced with compound B-9 to obtain compound B-128: 1 H NMR (300MHz, CDCl 3 )δ9.23(s,1H),8.77(d,J= 4.0Hz,1H), 8.29(d,J=8.0Hz,1H),7.42–7.35(m,1H),5.32–5.27(m,1H),4.82–4.72(m,1H),4.18–4.02(m ,4H),3.14–2.91(m,5H),2.30–2.13(m,1H),1.17(s,3H),1.02(s,3H),0.98(s,3H),0.94(s,6H), 0.92 (s, 3H), 0.74 (s, 3H). ESI-MS: m/z 679.5 [M+H] + .
实施例168Example 168
N-(3β-(3-羧基丙酰)氧基-齐墩果烷-12-烯-28-酰基)-硫代吗啉-1,1-二氧化物(化合物B-129)N-(3β-(3-carboxypropionyl)oxy-oleanorane-12-ene-28-acyl)-thiomorpholine-1,1-dioxide (compound B-129)
Figure PCTCN2020093248-appb-000208
Figure PCTCN2020093248-appb-000208
参照实施例127的方法，将化合物B-2替换成化合物B-9，制得化合物B-129： 1H NMR(300MHz,DMSO-d 6)δ12.19(s,1H),5.14–5.06(m,1H),4.45–4.35(m,1H),4.01–3.84(m,4H),3.15–2.88(m,5H),1.10(s,3H),0.88(s,9H),0.81(s,6H),0.66(s,3H).ESI-MS:m/z 674.5[M+H] +。 According to the method of Example 127, compound B-2 was replaced with compound B-9 to obtain compound B-129: 1 H NMR (300MHz, DMSO-d 6 ) δ 12.19 (s, 1H), 5.14–5.06 ( m,1H),4.45-4.35(m,1H),4.01-3.84(m,4H),3.15-2.88(m,5H),1.10(s,3H),0.88(s,9H),0.81(s, 6H), 0.66(s, 3H). ESI-MS: m/z 674.5 [M+H] + .
实施例169Example 169
N-(3β-(2S-氨基-3-甲基丁酰)氧基-齐墩果烷-12-烯-28-酰基)-硫代吗啉-1,1-二氧化物盐酸盐(化合物B-130)N-(3β-(2S-amino-3-methylbutyryl)oxy-oleanorane-12-ene-28-acyl)-thiomorpholine-1,1-dioxide hydrochloride ( Compound B-130)
Figure PCTCN2020093248-appb-000209
Figure PCTCN2020093248-appb-000209
参照实施例159的方法，将化合物B-20替换成化合物B-9，制得化合物B-130： 1H NMR(300MHz,DMSO-d 6)δ8.53–8.37(m,3H),5.15–5.06(m,1H),4.60–4.50(m,1H),4.02–3.82(m,5H),3.18–2.87(m,5H),1.11(s,3H),1.01(s,3H),0.99(s,3H),0.90(s,3H),0.87(s,6H),0.66(s,3H).ESI-MS:m/z 671.5[M+H] +。 According to the method of Example 159, compound B-20 was replaced with compound B-9 to prepare compound B-130: 1 H NMR (300MHz, DMSO-d 6 ) δ8.53–8.37(m,3H), 5.15– 5.06(m,1H), 4.60-4.50(m,1H), 4.02-3.82(m,5H), 3.18-2.87(m,5H), 1.11(s,3H), 1.01(s,3H), 0.99( s, 3H), 0.90 (s, 3H), 0.87 (s, 6H), 0.66 (s, 3H). ESI-MS: m/z 671.5 [M+H] + .
实施例170Example 170
3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酸3-(4-吗啉基)丙酯(化合物B-131)3β-(2-(2-Acetylaminoethoxyyl)benzoyl)oxy-oleanorane-12-en-28-acid 3-(4-morpholinyl)propyl ester (Compound B-131)
Figure PCTCN2020093248-appb-000210
Figure PCTCN2020093248-appb-000210
参照实施例136的方法，将1,2-二溴乙烷替换成1,3-二溴丙烷，制得化合物B-131： 1H NMR(300MHz,CDCl 3)δ7.85–7.79(m,1H),7.69–7.63(m,1H),7.62–7.51(m,2H),6.60–6.50(m,1H),5.32–5.26(m,1H),4.75–4.66(m,1H),4.48–4.39(m,2H),4.13–4.01(m,2H),3.75–3.68(m,4H),3.68–3.59(m,2H),2.87(d,J＝10.3Hz,1H),2.52–2.31(m,6H),2.02(s,3H),1.15(s,3H),0.97(s,9H),0.93(s,3H),0.91(s,3H),0.75(s,3H).ESI-MS:m/z 817.7[M+H] +。 According to the method of Example 136, 1,2-dibromoethane was replaced with 1,3-dibromopropane to obtain compound B-131: 1 H NMR (300MHz, CDCl 3 ) δ7.85-7.79(m, 1H), 7.69–7.63(m,1H), 7.62–7.51(m,2H), 6.60–6.50(m,1H), 5.32–5.26(m,1H), 4.75–4.66(m,1H), 4.48– 4.39 (m, 2H), 4.13-4.01 (m, 2H), 3.75-3.68 (m, 4H), 3.68-3.59 (m, 2H), 2.87 (d, J = 10.3 Hz, 1H), 2.52-2.31 ( m,6H),2.02(s,3H),1.15(s,3H),0.97(s,9H),0.93(s,3H),0.91(s,3H),0.75(s,3H).ESI-MS :m/z 817.7[M+H] + .
实施例171Example 171
3β-(2-(2-乙酰氨基乙氧酰基)苯甲酰基)氧基-齐墩果烷-12-烯-28-酸3-(1,1-二 氧代硫代吗啉基)丙酯(化合物B-132)3β-(2-(2-Acetylaminoethoxyacyl)benzoyl)oxy-oleanorane-12-ene-28-acid 3-(1,1-dioxothiomorpholinyl)propane Ester (Compound B-132)
Figure PCTCN2020093248-appb-000211
Figure PCTCN2020093248-appb-000211
参照实施例161的方法，将化合物XI-1替换成化合物XVII-4，制得化合物B-132： 1H NMR(300MHz,CDCl 3)δ7.89–7.77(m,1H),7.73–7.63(m,1H),7.62–7.50(m,2H),6.60–6.45(m,1H),5.34–5.24(m,1H),4.75–4.64(m,1H),4.52–4.36(m,2H),4.14–4.01(m,2H),3.73–3.54(m,2H),3.02(d,J＝13.2Hz,8H),2.86(d,J＝14.1Hz,1H),2.65–2.51(m,2H),1.16(s,3H),0.97(s,9H),0.92(s,6H),0.75(s,3H).ESI-MS:m/z 865.6[M+H] +。 According to the method of Example 161, compound XI-1 was replaced with compound XVII-4 to obtain compound B-132: 1 H NMR (300MHz, CDCl 3 )δ7.89-7.77(m,1H),7.73-7.63( m,1H), 7.62-7.50(m,2H), 6.60-6.45(m,1H), 5.34-5.24(m,1H), 4.75-4.64(m,1H), 4.52--4.36(m,2H), 4.14–4.01(m,2H), 3.73–3.54(m,2H), 3.02(d,J=13.2Hz,8H), 2.86(d,J=14.1Hz,1H), 2.65–2.51(m,2H) , 1.16 (s, 3H), 0.97 (s, 9H), 0.92 (s, 6H), 0.75 (s, 3H). ESI-MS: m/z 865.6 [M+H] + .
实施例172Example 172
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-苄基-哌嗪(化合物B-133)N-(3β-Hydroxy-oleanan-12-ene-28-acyl)-4-benzyl-piperazine (Compound B-133)
Figure PCTCN2020093248-appb-000212
Figure PCTCN2020093248-appb-000212
参照实施例73的方法，将N,N-二甲基乙二胺替换成1-苄基哌嗪，制得化合物B-133： 1H NMR(300MHz,CDCl 3)δ7.39–7.27(m,5H),5.28–5.21(m,1H),3.78–3.60(m,4H),3.57(s,2H),3.27–3.16(m,1H),3.14–3.01(m,1H),2.58–2.31(m,4H),2.16–2.02(m,1H),1.13(s,3H),0.99(s,3H),0.92(s,3H),0.90(s,3H),0.89(s,3H),0.78(s,3H),0.71(s,3H).ESI-MS:m/z 615.6[M+H] +。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with 1-benzylpiperazine to obtain compound B-133: 1 H NMR (300MHz, CDCl 3 )δ7.39-7.27(m ,5H), 5.28–5.21(m,1H), 3.78–3.60(m,4H), 3.57(s,2H), 3.27–3.16(m,1H), 3.14–3.01(m,1H), 2.58–2.31 (m,4H),2.16-2.02(m,1H),1.13(s,3H),0.99(s,3H),0.92(s,3H),0.90(s,3H),0.89(s,3H), 0.78 (s, 3H), 0.71 (s, 3H). ESI-MS: m/z 615.6 [M+H] + .
实施例173Example 173
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-环丙基-哌嗪(化合物B-134)N-(3β-hydroxy-oleanane-12-ene-28-acyl)-4-cyclopropyl-piperazine (compound B-134)
Figure PCTCN2020093248-appb-000213
Figure PCTCN2020093248-appb-000213
参照实施例73的方法，将N,N-二甲基乙二胺替换成1-环丙基哌嗪，制得化合物B-134： 1H NMR(300MHz,CDCl 3)δ5.32–5.21(m,1H),3.72–3.44(m,4H),3.27–3.15(m,1H),3.15–3.03(m,1H),2.70–2.45(m,4H),2.19–2.03(m,1H),1.14(s,3H),0.99(s,3H),0.93(s,3H),0.90(s,3H),0.90(s,3H),0.78(s,3H),0.75(s,3H).ESI-MS:m/z 565.6[M+H] +。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with 1-cyclopropylpiperazine to obtain compound B-134: 1 H NMR (300MHz, CDCl 3 )δ5.32-5.21( m,1H), 3.72–3.44(m,4H), 3.27–3.15(m,1H), 3.15–3.03(m,1H), 2.70–2.45(m,4H), 2.19–2.03(m,1H), 1.14(s,3H),0.99(s,3H),0.93(s,3H),0.90(s,3H),0.90(s,3H),0.78(s,3H),0.75(s,3H).ESI -MS: m/z 565.6 [M+H] + .
实施例174Example 174
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-乙基-哌嗪(化合物B-135)N-(3β-hydroxy-oleanane-12-ene-28-acyl)-4-ethyl-piperazine (compound B-135)
Figure PCTCN2020093248-appb-000214
Figure PCTCN2020093248-appb-000214
参照实施例73的方法，将N,N-二甲基乙二胺替换成N-乙基哌嗪，制得化合物B-135： 1H NMR(300MHz,CDCl 3)δ5.32–5.23(m,1H),3.77–3.58(m,4H),3.29–3.16(m,1H),3.16–3.02(m,1H),2.54–2.30(m,6H),2.20–2.03(m,1H),1.14(s,3H),0.99(s,3H),0.93(s,3H),0.90(s,6H),0.78(s,3H),0.74(s,3H).ESI-MS:m/z 553.6[M+H] +。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with N-ethylpiperazine to obtain compound B-135: 1 H NMR (300MHz, CDCl 3 )δ5.32–5.23(m ,1H),3.77–3.58(m,4H), 3.29–3.16(m,1H), 3.16–3.02(m,1H), 2.54–2.30(m,6H), 2.20–2.03(m,1H), 1.14 (s,3H),0.99(s,3H),0.93(s,3H),0.90(s,6H),0.78(s,3H),0.74(s,3H).ESI-MS:m/z 553.6[ M+H] + .
实施例175Example 175
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-氧代-哌啶(化合物B-136)N-(3β-Hydroxy-oleanane-12-ene-28-acyl)-4-oxo-piperidine (Compound B-136)
Figure PCTCN2020093248-appb-000215
Figure PCTCN2020093248-appb-000215
参照实施例73的方法，将N,N-二甲基乙二胺替换成4-哌啶酮，制得化合物B-136： 1H NMR(300MHz,CDCl 3)δ5.33–5.26(m,1H),3.98–3.79(m,4H),3.26–3.16(m,1H),3.15–3.04(m,1H),2.56–2.33(m,4H),2.27–2.11(m,1H),1.16(s,3H),0.99(s,3H),0.95(s,3H),0.91(s,3H),0.91(s,3H),0.78(s,3H),0.75(s,3H).ESI-MS:m/z 560.5[M+Na] +。 Referring to the method in Example 73, N,N-dimethylethylenediamine was replaced with 4-piperidone to obtain compound B-136: 1 H NMR (300MHz, CDCl 3 ) δ5.33-5.26 (m, 1H), 3.98–3.79(m,4H), 3.26–3.16(m,1H), 3.15–3.04(m,1H), 2.56–2.33(m,4H), 2.27–2.11(m,1H), 1.16( s,3H),0.99(s,3H),0.95(s,3H),0.91(s,3H),0.91(s,3H),0.78(s,3H),0.75(s,3H).ESI-MS :m/z 560.5[M+Na] + .
实施例176Example 176
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-羟基-哌啶(化合物B-137)N-(3β-hydroxy-oleanorane-12-ene-28-acyl)-4-hydroxy-piperidine (compound B-137)
Figure PCTCN2020093248-appb-000216
Figure PCTCN2020093248-appb-000216
参照实施例73的方法，将N,N-二甲基乙二胺替换成4-羟基哌啶，制得化合物B-137： 1H NMR(300MHz,CDCl 3)δ5.30–5.23(m,1H),4.18–3.99(m,2H),3.97–3.84(m,1H),3.28–3.01(m,4H),1.14(s,3H),0.99(s,3H),0.93(s,3H),0.90(s,6H),0.78(s,3H),0.75(s,3H).ESI-MS:m/z 540.5[M+H] +。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with 4-hydroxypiperidine to obtain compound B-137: 1 H NMR (300MHz, CDCl 3 )δ5.30–5.23(m, 1H), 4.18-3.99(m, 2H), 3.97-3.84(m, 1H), 3.28-3.01(m, 4H), 1.14(s, 3H), 0.99(s, 3H), 0.93(s, 3H) , 0.90 (s, 6H), 0.78 (s, 3H), 0.75 (s, 3H). ESI-MS: m/z 540.5 [M+H] + .
实施例177Example 177
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-3-羟甲基-吖丁啶(化合物B-138)N-(3β-Hydroxy-oleanorane-12-ene-28-acyl)-3-hydroxymethyl-azetidine (Compound B-138)
Figure PCTCN2020093248-appb-000217
Figure PCTCN2020093248-appb-000217
参照实施例73的方法，将N,N-二甲基乙二胺替换成3-氮杂丁烷甲醇，制得化合物B-138： 1H NMR(300MHz,CDCl 3)δ5.26(t,J＝3.0Hz,1H),4.54–3.82(m,4H),3.81–3.70(m,2H),3.27–3.14(m,1H),2.86(dd,J＝13.6,3.1Hz,1H),2.79–2.63(m,1H),1.13(s,3H),0.99(s,3H),0.93(s,3H),0.91(s,3H),0.89(s,3H),0.78(s,3H),0.77(s,3H).ESI-MS:m/z 524.5[M-H] -。 Referring to the method in Example 73, N,N-dimethylethylenediamine was replaced with 3-azetidine methanol to obtain compound B-138: 1 H NMR (300MHz, CDCl 3 )δ 5.26(t, J = 3.0Hz, 1H), 4.54–3.82 (m, 4H), 3.81–3.70 (m, 2H), 3.27–3.14 (m, 1H), 2.86 (dd, J = 13.6, 3.1 Hz, 1H), 2.79 --2.63(m,1H),1.13(s,3H),0.99(s,3H),0.93(s,3H),0.91(s,3H),0.89(s,3H),0.78(s,3H), 0.77(s, 3H). ESI-MS: m/z 524.5[MH] - .
实施例178Example 178
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-3-甲氧甲酰基-吖丁啶(化合物B-139)N-(3β-Hydroxy-oleanan-12-ene-28-acyl)-3-methoxyformyl-azetidine (Compound B-139)
Figure PCTCN2020093248-appb-000218
Figure PCTCN2020093248-appb-000218
参照实施例73的方法，将N,N-二甲基乙二胺替换成3-甲酸甲酯氮杂环丁烷盐酸盐，制得化合物B-139： 1H NMR(300MHz,CDCl 3)δ5.38–5.19(m,1H),4.69–4.00(m,4H),3.75(s,3H),3.42–3.29(m,1H),3.21(dd,J＝9.1,4.7Hz,1H),2.82(dd,J＝13.8,3.5Hz,1H),1.13(s,3H),0.99(s,3H),0.93(s,3H),0.91(s,3H),0.90(s,3H),0.79(s,3H),0.76(s,3H).ESI-MS:m/z 552.5[M-H] -。 According to the method of Example 73, N,N-dimethylethylenediamine was replaced with methyl 3-formate azetidine hydrochloride to obtain compound B-139: 1 H NMR (300MHz, CDCl 3 ) δ5.38–5.19(m,1H), 4.69–4.00(m,4H), 3.75(s,3H), 3.42–3.29(m,1H), 3.21(dd,J=9.1,4.7Hz,1H), 2.82(dd,J=13.8,3.5Hz,1H),1.13(s,3H),0.99(s,3H),0.93(s,3H),0.91(s,3H),0.90(s,3H),0.79 (s, 3H), 0.76 (s, 3H). ESI-MS: m/z 552.5 [MH] - .
实施例179Example 179
N-(3β-(吡啶-3-甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-3-甲氧基-吖丁啶(化合物B-140)N-(3β-(pyridine-3-formyl)oxy-oleanorane-12-en-28-acyl)-3-methoxy-azetidine (compound B-140)
Figure PCTCN2020093248-appb-000219
Figure PCTCN2020093248-appb-000219
参照实施例158的方法，将化合物B-20替换成化合物B-110，制得化合物B-140： 1H NMR(300MHz,CDCl 3)δ9.23(s,1H),8.77(d,J＝3.8Hz,1H),8.37–8.23(m,1H),7.39(dd,J＝7.9,4.9Hz,1H),5.27(t,J＝2.9Hz,1H),4.86–4.70(m,2H),4.44–3.86(m,5H),3.30(s,3H),2.87(dd,J＝13.5,3.6Hz,1H),1.15(s,3H),1.02(s,3H),0.99(s,3H),0.95(s,3H),0.93(s,3H),0.90(s,3H),0.78(s,3H).ESI-MS:m/z 631.5[M+H] +。 According to the method of Example 158, compound B-20 was replaced with compound B-110 to obtain compound B-140: 1 H NMR (300MHz, CDCl 3 ) δ 9.23 (s, 1H), 8.77 (d, J = 3.8Hz,1H), 8.37–8.23(m,1H), 7.39(dd,J=7.9,4.9Hz,1H), 5.27(t,J=2.9Hz,1H), 4.86–4.70(m,2H), 4.44–3.86 (m, 5H), 3.30 (s, 3H), 2.87 (dd, J = 13.5, 3.6 Hz, 1H), 1.15 (s, 3H), 1.02 (s, 3H), 0.99 (s, 3H) , 0.95 (s, 3H), 0.93 (s, 3H), 0.90 (s, 3H), 0.78 (s, 3H). ESI-MS: m/z 631.5 [M+H] + .
实施例180Example 180
N-(3β-(吡啶-2-甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-3-甲氧基-吖丁啶(化合 物B-141)N-(3β-(pyridine-2-formyl)oxy-oleanorane-12-en-28-acyl)-3-methoxy-azetidine (compound B-141)
Figure PCTCN2020093248-appb-000220
Figure PCTCN2020093248-appb-000220
参照实施例158的方法，将化合物B-20替换成化合物B-110，烟酸替换成2-吡啶甲酸，制得化合物B-141： 1H NMR(300MHz,CDCl 3)δ8.83–8.74(m,1H),8.06(d,J＝7.8Hz,1H),7.82(td,J＝7.7,1.6Hz,1H),7.49–7.40(m,1H),5.27(d,J＝2.9Hz,1H),4.93–4.80(m,1H),4.45–3.81(m,5H),3.30(s,3H),2.93–2.79(m,1H),1.15(s,3H),1.04(s,3H),0.99(s,3H),0.96(s,3H),0.93(s,3H),0.90(s,3H),0.78(s,3H).ESI-MS:m/z 631.5[M+H] +。 According to the method in Example 158, compound B-20 was replaced with compound B-110, and niacin was replaced with 2-picolinic acid to prepare compound B-141: 1 H NMR (300MHz, CDCl 3 ) δ 8.83–8.74 ( m, 1H), 8.06 (d, J = 7.8 Hz, 1H), 7.82 (td, J = 7.7, 1.6 Hz, 1H), 7.49-7.40 (m, 1H), 5.27 (d, J = 2.9 Hz, 1H ), 4.93–4.80(m,1H), 4.45–3.81(m,5H), 3.30(s,3H), 2.93–2.79(m,1H), 1.15(s,3H), 1.04(s,3H), 0.99 (s, 3H), 0.96 (s, 3H), 0.93 (s, 3H), 0.90 (s, 3H), 0.78 (s, 3H). ESI-MS: m/z 631.5 [M+H] + .
实施例181Example 181
N-(3β-(呋喃-2-甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-3-甲氧基-吖丁啶(化合物B-142)N-(3β-(furan-2-formyl)oxy-oleanorane-12-en-28-acyl)-3-methoxy-azetidine (compound B-142)
Figure PCTCN2020093248-appb-000221
Figure PCTCN2020093248-appb-000221
参照实施例158的方法，将化合物B-20替换成化合物B-110，烟酸替换成糠酸，制得化合物B-142： 1H NMR(300MHz,CDCl 3)δ8.06–7.98(m,1H),7.45(t,J＝1.7Hz,1H),6.81–6.73(m,1H),5.30(t,J＝3.2Hz,1H),4.70(dd,J＝9.3,6.8Hz,1H),4.55–3.86(m,5H),3.33(s,3H),2.89(dd,J＝13.3,3.2Hz,1H),1.18(s,3H),1.00(s,3H),0.98(s,3H),0.96(s,3H),0.95(s,3H),0.93(s,3H),0.81(s,3H).ESI-MS:m/z 631.5[M+H] +。 According to the method of Example 158, compound B-20 was replaced with compound B-110, and niacin was replaced with furoic acid to prepare compound B-142: 1 H NMR (300MHz, CDCl 3 ) δ8.06-7.98 (m, 1H),7.45(t,J=1.7Hz,1H),6.81-6.73(m,1H),5.30(t,J=3.2Hz,1H),4.70(dd,J=9.3,6.8Hz,1H), 4.55–3.86(m,5H),3.33(s,3H),2.89(dd,J=13.3,3.2Hz,1H),1.18(s,3H),1.00(s,3H),0.98(s,3H) , 0.96 (s, 3H), 0.95 (s, 3H), 0.93 (s, 3H), 0.81 (s, 3H). ESI-MS: m/z 631.5 [M+H] + .
实施例182Example 182
N-(3β-(6-氟吡啶-2-甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-3-甲氧基-吖丁啶(化合物B-143)N-(3β-(6-Fluoropyridine-2-formyl)oxy-oleanorane-12-en-28-acyl)-3-methoxy-azetidine (compound B-143)
Figure PCTCN2020093248-appb-000222
Figure PCTCN2020093248-appb-000222
参照实施例158的方法，将化合物B-20替换成化合物B-110，烟酸替换成2-氟吡啶-6-羧酸，制得化合物B-143： 1H NMR(300MHz,CDCl 3)δ8.00–7.87(m,2H),7.17–7.09(m,1H),5.30–5.24(m,1H),4.38–3.87(m,5H),3.30(s,3H),2.92–2.81(m,1H),1.15(s,3H),1.03(s,3H),0.98(s,3H),0.95(s,3H),0.93(s,3H),0.90 (s,3H),0.78(s,3H).ESI-MS:m/z 649.5[M+H] +。 According to the method of Example 158, compound B-20 was replaced with compound B-110, and nicotinic acid was replaced with 2-fluoropyridine-6-carboxylic acid to prepare compound B-143: 1 H NMR (300MHz, CDCl 3 )δ8 .00–7.87(m,2H),7.17–7.09(m,1H),5.30–5.24(m,1H), 4.38–3.87(m,5H), 3.30(s,3H), 2.92–2.81(m, 1H), 1.15 (s, 3H), 1.03 (s, 3H), 0.98 (s, 3H), 0.95 (s, 3H), 0.93 (s, 3H), 0.90 (s, 3H), 0.78 (s, 3H) ). ESI-MS: m/z 649.5 [M+H] + .
实施例183Example 183
N-(3β-(6-氯吡啶-2-甲酰基)氧基-齐墩果烷-12-烯-28-酰基)-3-甲氧基-吖丁啶(化合物B-144)N-(3β-(6-Chloropyridine-2-formyl)oxy-oleanorane-12-en-28-acyl)-3-methoxy-azetidine (compound B-144)
Figure PCTCN2020093248-appb-000223
Figure PCTCN2020093248-appb-000223
参照实施例158的方法，将化合物B-20替换成化合物B-110，烟酸替换成2-氯吡啶-6-羧酸，制得化合物B-144： 1H NMR(300MHz,CDCl 3)δ7.97(d,J＝7.5Hz,1H),7.78(t,J＝7.7Hz,1H),7.49(d,J＝7.9Hz,1H),5.30–5.24(m,1H),4.88–4.76(m,1H),4.43–3.85(m,5H),3.30(s,3H),2.92–2.81(m,1H),1.15(s,3H),1.02(s,3H),0.98(s,3H),0.94(s,3H),0.93(s,3H),0.90(s,3H),0.78(s,3H).ESI-MS:m/z 665.5[M+H] +。 According to the method of Example 158, compound B-20 was replaced with compound B-110, and niacin was replaced with 2-chloropyridine-6-carboxylic acid, to obtain compound B-144: 1 H NMR (300MHz, CDCl 3 )δ7 .97(d,J=7.5Hz,1H),7.78(t,J=7.7Hz,1H),7.49(d,J=7.9Hz,1H),5.30–5.24(m,1H),4.88–4.76( m,1H), 4.43--3.85(m,5H), 3.30(s,3H), 2.92--2.81(m,1H), 1.15(s,3H), 1.02(s,3H), 0.98(s,3H) , 0.94 (s, 3H), 0.93 (s, 3H), 0.90 (s, 3H), 0.78 (s, 3H). ESI-MS: m/z 665.5 [M+H] + .
实施例184Example 184
化合物对Huh-7细胞AMPK激动作用的活性评价Evaluation of the compound's agonistic activity on AMPK in Huh-7 cells
采用Western Blot方法检测化合物对Huh-7细胞AMPK的激动活性。The Western Blot method was used to detect the agonistic activity of the compound on Huh-7 cells AMPK.
1、细胞系：Huh-7细胞(人源肝癌细胞，购自中国科学院干细胞库)，用DMEM完全培养基(含10％胎牛血清和1％streptomycin/penicillin)，在37℃含有5％CO 2的细胞培养箱中培养。 1. Cell line: Huh-7 cells (human liver cancer cells, purchased from the Stem Cell Bank of the Chinese Academy of Sciences), using DMEM complete medium (containing 10% fetal bovine serum and 1% streptomycin/penicillin), containing 5% CO at 37°C 2. Cultivate in the cell incubator.
2、抗体：anti-AMPK(Cell Signaling Technology 2532S)；anti-pAMPK(Thr172，Cell Signaling Technology 2535S)；GAPDH(arigobio)，HRP标记的山羊抗兔IgG二抗，HRP标记的山羊抗小鼠IgG二抗(碧云天)。2. Antibody: anti-AMPK (Cell Signaling Technology 2532S); anti-pAMPK (Thr172, Cell Signaling Technology 2535S); GAPDH (arigobio), HRP-labeled goat anti-rabbit IgG secondary antibody, HRP-labeled goat anti-mouse IgG secondary antibody Anti (Biyuntian).
3、Western Blot实验：检测化合物对Huh-7细胞AMPK磷酸化水平的影响。3. Western Blot experiment: to detect the effect of the compound on the AMPK phosphorylation level of Huh-7 cells.
取活细胞比例90％以上的细胞进行实验。在12孔板中，Huh-7细胞按照20万每孔铺板。在37℃含有5％CO 2的培养箱中培养24小时，贴壁。在完全培养基条件下给予受试化合物，化合物终浓度均设置为10μM，给药时间为12小时，采用AICAR(100μM)作为阳性对照化合物。随后提取蛋白进行Western Blot检测。具体步骤如下： Take more than 90% of live cells for experiment. In a 12-well plate, Huh-7 cells are plated at 200,000 per well. Incubate for 24 hours in an incubator containing 5% CO 2 at 37°C, and adhere to the wall. The test compound was administered under the condition of complete medium, the final concentration of the compound was set to 10 μM, the administration time was 12 hours, and AICAR (100 μM) was used as the positive control compound. The protein was then extracted for Western Blot detection. Specific steps are as follows:
蛋白样品制备：弃去原培养液，用1×PBS洗涤3次，弃去PBS，每孔加入100μl RIPA buffer(1×PBS，1％NP40，0.5％脱氧胆酸钠，0.1％SDS，PMSF等)，冰上孵育15min，用细胞刮刀将细胞刮下，吸至新的1.5ml EP管中，4℃，12000g，离心15min，将上清转移至新的1.5ml EP管中，放置于冰上，加入1/5体积的6×loading buffer，95℃金属浴加热10min后离心1min，冻存至-20℃备用。Protein sample preparation: discard the original culture medium, wash 3 times with 1×PBS, discard PBS, add 100μl RIPA buffer (1×PBS, 1% NP40, 0.5% sodium deoxycholate, 0.1% SDS, PMSF, etc.) to each well ), incubate on ice for 15 minutes, scrape the cells with a cell scraper, suck into a new 1.5ml EP tube, 4℃, 12000g, centrifuge for 15min, transfer the supernatant to a new 1.5ml EP tube, and place on ice , Add 1/5 volume of 6×loading buffer, heat in a metal bath at 95℃ for 10min, centrifuge for 1min, freeze and store at -20℃ for later use.
蛋白定量：将蛋白样品20倍稀释，在96孔板中，依次加入20μl稀释后的蛋白样品，200μl BCA(A液：B液＝5：1)试剂，37℃孵育30min，在酶标仪上562nm波长下测定OD值，根据标准曲线计算蛋白浓度。Protein quantification: Dilute the protein sample 20 times, add 20μl diluted protein sample, 200μl BCA (A solution: B solution = 5:1) reagent to a 96-well plate, incubate at 37°C for 30 minutes, and place it on the microplate reader Measure the OD value at 562nm wavelength, and calculate the protein concentration according to the standard curve.
电泳：灌制SDS-PAGE胶，分离胶浓度为10％，浓缩胶浓度为4％。蛋白样品预先在95℃金属浴加热4min后离心1min，每个样品上样30μg总蛋白，用微量进样器逐个加样。连接电源(注意正负极连接)，开始时以60V恒压电泳，当蛋白样品进入分离胶时将电压调至100V继续恒压电泳。当溴酚蓝到达分离胶底部时，根据蛋白marker的分离情况终止电泳。Electrophoresis: Pouring SDS-PAGE gel, separation gel concentration is 10%, concentrated gel concentration is 4%. The protein samples were heated in a metal bath at 95°C for 4 minutes and then centrifuged for 1 minute. Each sample was loaded with 30 μg total protein, and the samples were added one by one with a micro-injector. Connect the power supply (note that the positive and negative poles are connected), start electrophoresis at a constant voltage of 60V, when the protein sample enters the separation gel, adjust the voltage to 100V to continue the constant voltage electrophoresis. When bromophenol blue reaches the bottom of the separation gel, the electrophoresis is terminated according to the separation of the protein marker.
转膜：将电泳后的胶轻轻取出，切去不需要的部分，将需要的胶浸入Transfer buffer。准备与胶的大小一致的PVDF膜，用前将PVDF膜用甲醇浸透1min，转移至Transfer buffer中，同时将滤纸放入Transfer buffer中浸透。在转印仪阴极的电极板上依次铺上海绵、滤纸、胶、PVDF膜、滤纸和海绵。每层之间都避免有气泡。接通电源，200mA恒流冰浴转印2.5h。Transfer membrane: Gently take out the gel after electrophoresis, cut off the unnecessary part, and immerse the required gel into the Transfer buffer. Prepare a PVDF membrane with the same size as the glue, soak the PVDF membrane with methanol for 1 min before use, transfer it to the Transfer buffer, and put the filter paper into the Transfer buffer to soak. Spread sponge, filter paper, glue, PVDF membrane, filter paper, and sponge on the electrode plate of the cathode of the transfer apparatus. Avoid air bubbles between each layer. Turn on the power supply and transfer in a 200mA constant current ice bath for 2.5 hours.
抗体杂交：转膜结束后，取出PVDF膜用l×TBST清洗1遍，放入预先准备好的封闭液中(含0.1％Tween20的1×TBS配置的5％BSA溶液)，室温封闭1h。一抗孵育4℃过夜。第二天(12h后)，用1×TBST洗3次，每次10min。二抗孵育：用5％BSA稀释的二抗(1：10000)室温孵育1h，1×TBST洗3次，每次10min。吸去PVDF膜上的多余液体，铺在曝光板上，加入等体积混合的ECL试剂盒液体Tanon TM High-sig ECL Western Blotting Peroxide Buffer和Tanon TM High-sig ECL Western Blotting Luminol/Enhancer Solution，运用Tanon化学发光成像仪,ECL显影,采集免疫反应带。 Antibody hybridization: After the transfer, take out the PVDF membrane and wash it with 1×TBST once, put it in the pre-prepared blocking solution (1×TBS with 0.1% Tween20 in 5% BSA solution), and block at room temperature for 1 hour. Incubate the primary antibody at 4°C overnight. On the second day (after 12h), wash 3 times with 1×TBST, each time for 10 minutes. Secondary antibody incubation: Incubate with 5% BSA diluted secondary antibody (1:10000) for 1 hour at room temperature, wash 3 times with 1×TBST, 10 minutes each time. Aspirate the excess liquid on the PVDF membrane, spread it on the exposure plate, and add an equal volume of ECL kit liquid Tanon TM High-sig ECL Western Blotting Peroxide Buffer and Tanon TM High-sig ECL Western Blotting Luminol/Enhancer Solution, using Tanon Chemiluminescence imager, ECL development, collection of immune response zone.
4.、实验结果：对Western Blot实验结果进行灰度扫描后将阴性对照DMSO的p-AMPK/AMPK比值定义为1，本发明合成的受试化合物p-AMPK/AMPK比值为阴性对照组的相对比值，该数值越大，表明化合物的AMPK激动活性越强，活性数据结果如表1所示。4. Experimental results: After performing grayscale scanning on the results of the Western Blot experiment, the p-AMPK/AMPK ratio of the negative control DMSO is defined as 1, and the p-AMPK/AMPK ratio of the test compound synthesized in the present invention is the relative value of the negative control group. The larger the ratio, the greater the AMPK agonistic activity of the compound. The activity data results are shown in Table 1.
表1、化合物的AMPK激动活性(阳性对照AICAR浓度为100μM；受试化合物浓度为10μM)Table 1. AMPK agonistic activity of the compounds (positive control AICAR concentration is 100 μM; test compound concentration is 10 μM)
Figure PCTCN2020093248-appb-000224
Figure PCTCN2020093248-appb-000224
Figure PCTCN2020093248-appb-000225
Figure PCTCN2020093248-appb-000225
Figure PCTCN2020093248-appb-000226
Figure PCTCN2020093248-appb-000226
Figure PCTCN2020093248-appb-000227
Figure PCTCN2020093248-appb-000227
Figure PCTCN2020093248-appb-000228
Figure PCTCN2020093248-appb-000228
Figure PCTCN2020093248-appb-000229
Figure PCTCN2020093248-appb-000229
Figure PCTCN2020093248-appb-000230
Figure PCTCN2020093248-appb-000230
Figure PCTCN2020093248-appb-000231
Figure PCTCN2020093248-appb-000231
Figure PCTCN2020093248-appb-000232
Figure PCTCN2020093248-appb-000232
Figure PCTCN2020093248-appb-000233
Figure PCTCN2020093248-appb-000233
Figure PCTCN2020093248-appb-000234
Figure PCTCN2020093248-appb-000234
Figure PCTCN2020093248-appb-000235
Figure PCTCN2020093248-appb-000235
Figure PCTCN2020093248-appb-000236
Figure PCTCN2020093248-appb-000236
Figure PCTCN2020093248-appb-000237
Figure PCTCN2020093248-appb-000237
Figure PCTCN2020093248-appb-000238
Figure PCTCN2020093248-appb-000238
Figure PCTCN2020093248-appb-000239
Figure PCTCN2020093248-appb-000239
Figure PCTCN2020093248-appb-000240
Figure PCTCN2020093248-appb-000240
Figure PCTCN2020093248-appb-000241
Figure PCTCN2020093248-appb-000241
Figure PCTCN2020093248-appb-000242
Figure PCTCN2020093248-appb-000242
Figure PCTCN2020093248-appb-000243
Figure PCTCN2020093248-appb-000243
Figure PCTCN2020093248-appb-000244
Figure PCTCN2020093248-appb-000244
如表1的实验结果所示，在10μM浓度下，本发明提供的新型齐墩果酸和δ-齐墩果酸的衍生物具有显著的AMPK激动活性。例如，化合物A-9、A-12、A-23、A-25、A-27、A-28、A-68、A-81、A-85、A-108、A-109、A-110、A-112、B-9、B-20、B-63、B-65、B-80、B-94、B-110、B-121、B-122、B-123、B-125等化合物均是强效的AMPK激动剂，且活性显著优于100μM的AICAR以及齐墩果酸和δ-齐墩果酸。其中，化合物A-68的AMPK激动活性(pAMPK/AMPK＝5.48)是δ-齐墩果酸活性(pAMPK/AMPK＝1.45)的3倍以上。以上实验结果表明，本发明的化合物对AMPK具有显著的激动活性，因而可用于制备具有增强AMPK磷酸化水平活性的AMPK激动剂，并进而可用于制备预防或治疗AMPK介导的疾病的药物。本发明实施例中其他未列举化合物其合成方法均可参考上述实施例，并且对AMPK也同样具有显著的激动活性。As shown in the experimental results in Table 1, at a concentration of 10 μM, the novel derivatives of oleanolic acid and delta-oleanolic acid provided by the present invention have significant AMPK agonistic activity. For example, compounds A-9, A-12, A-23, A-25, A-27, A-28, A-68, A-81, A-85, A-108, A-109, A-110 , A-112, B-9, B-20, B-63, B-65, B-80, B-94, B-110, B-121, B-122, B-123, B-125 and other compounds Both are potent AMPK agonists, and their activity is significantly better than 100μM AICAR, oleanolic acid and delta-oleanolic acid. Among them, the AMPK agonistic activity of compound A-68 (pAMPK/AMPK=5.48) is more than 3 times that of δ-oleanolic acid (pAMPK/AMPK=1.45). The above experimental results show that the compound of the present invention has significant agonistic activity on AMPK, and therefore can be used to prepare AMPK agonists with the activity of enhancing the phosphorylation level of AMPK, and further can be used to prepare drugs for preventing or treating AMPK-mediated diseases. The synthesis methods of other compounds not listed in the examples of the present invention can be referred to the foregoing examples, and they also have significant agonistic activity on AMPK.
实施例185Example 185
化合物对Huh-7细胞AMPK下游信号通路的影响Effects of compounds on the downstream signal pathway of AMPK in Huh-7 cells
采用Western Blot方法检测化合物对Huh-7细胞AMPK下游信号通路的影响。Western Blot method was used to detect the effect of the compound on the downstream signal pathway of AMPK in Huh-7 cells.
细胞培养条件：Huh-7细胞：DMEM完全培养基(含10％胎牛血清和1％streptomycin/penicillin)在5％CO 2的37℃培养箱中培养。 Cell culture conditions: Huh-7 cells: DMEM complete medium (containing 10% fetal bovine serum and 1% streptomycin/penicillin) cultured in a 5% CO 2 incubator at 37°C.
抗体：anti-ACC(CST,3676S)；anti-pACC(CST,11818S)；anti-mTOR(CST,2983S)；anti-pmTOR(CST,Ser2448)。Antibodies: anti-ACC (CST, 3676S); anti-pACC (CST, 11818S); anti-mTOR (CST, 2983S); anti-pmTOR (CST, Ser2448).
Western Blot实验：检测化合物对Huh-7细胞AMPK下游信号通路的影响：Western Blot experiment: To detect the effect of compounds on the downstream signal pathway of AMPK in Huh-7 cells:
取活细胞比例90％以上的细胞进行实验。在12孔板中，将Huh-7细胞按照25万每孔铺板，置于含5％CO 2的37℃培养箱中培养，12小时后，弃去原先培养基，加入含有化合物的完全培养基，受试化合物的终浓度均设置为10μM，给药时间为12小时。阳性对照化合物采用AICAR(100μM)。随后提取蛋白进行Western Blot检测，检测pACC/ACC和pmTOR/mTOR的蛋白变化情况。实验结 果如图1所示。 Take more than 90% of live cells for experiment. In a 12-well plate, plate Huh-7 cells at a rate of 250,000 per well and place them in a 37°C incubator containing 5% CO 2 for culture. After 12 hours, discard the original medium and add complete medium containing the compound , The final concentration of the test compound was set to 10 μM, and the administration time was 12 hours. AICAR (100μM) was used as the positive control compound. Subsequently, the protein was extracted for Western Blot detection to detect the protein changes of pACC/ACC and pmTOR/mTOR. The experimental results are shown in Figure 1.
实验结果(图1)表明，本发明的化合物均能有效地引起AMPK下游蛋白的变化，且其变化趋势与AMPK激动相关。例如，化合物B-125、A-112、B-122、A-109、B-121、A-108、B-123、A-110、B-20和A-81等均能增加AMPK的磷酸化水平，降低mTOR的磷酸化水平，并同时增加ACC的磷酸化水平。此外，本发明其他化合物均能增加AMPK的磷酸化水平，降低mTOR的磷酸化水平，并同时增加ACC的磷酸化水平。The experimental results (Figure 1) show that the compounds of the present invention can effectively cause changes in AMPK downstream proteins, and the change trend is related to AMPK activation. For example, compounds B-125, A-112, B-122, A-109, B-121, A-108, B-123, A-110, B-20 and A-81 can all increase the phosphorylation of AMPK Level, reduce the phosphorylation level of mTOR and increase the phosphorylation level of ACC at the same time. In addition, other compounds of the present invention can increase the phosphorylation level of AMPK, reduce the phosphorylation level of mTOR, and increase the phosphorylation level of ACC at the same time.
实施例186Example 186
片剂tablet
将实施例41中制得的化合物A-81或者其他实施例的化合物(50g)、羟丙甲基纤维素E(150g)、淀粉(200g)、聚维酮K30适量和硬脂酸镁(1g)混合，制粒，压片。The compound A-81 prepared in Example 41 or the compound of other examples (50g), hydroxypropylmethyl cellulose E (150g), starch (200g), a proper amount of povidone K30 and magnesium stearate (1g) ) Mixing, granulating, and tableting.
此外，可以根据药典2015版常规制剂法，将实施例1～183制得的化合物赋予不同的药物辅料制成胶囊剂、散剂、颗粒剂、丸剂、注射剂、糖浆剂、口服液、吸入剂、软膏剂、栓剂或贴剂等。In addition, according to the pharmacopoeia 2015 version of the conventional preparation method, the compounds prepared in Examples 1 to 183 can be given different pharmaceutical excipients into capsules, powders, granules, pills, injections, syrups, oral liquids, inhalants, ointments Agent, suppository or patch, etc.

Claims (10)

  1. 如下式I或式II所示的δ-齐墩果酸衍生物或齐墩果酸衍生物、其药学上可接受的盐或酯或溶剂化物：Delta-oleanolic acid derivatives or oleanolic acid derivatives, pharmaceutically acceptable salts or esters or solvates thereof as shown in the following formula I or formula II:
    Figure PCTCN2020093248-appb-100001
    Figure PCTCN2020093248-appb-100001
    R是H、
    Figure PCTCN2020093248-appb-100002
    或R aCO-；     R is H,
    Figure PCTCN2020093248-appb-100002
    Or R a CO-;
    R 1是H、C 1-C 5烷基或取代基Y取代的C 1-C 5烷基，所述取代基Y是OH、C(O)OH、C(O)NH 2、NH 2、NHC(O)CH 3、吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基或二乙醇胺基； R 1 is H, C 1 -C 5 alkyl substituted by substituent Y or a C 1 -C 5 alkyl, the substituent Y is OH, C (O) OH, C (O) NH 2, NH 2, NHC(O)CH 3 , pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorph Lin-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethylammonium or diethanolamino;
    R a是非取代的或取代基L取代的C 1-C 5烷基，所述取代基L是一个或两个独立地选自下列的取代基：OH、C(O)OH、NH 2、吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基或NHC(O)CH(CH 3)NHC(O)CH(CH 3)NH； R a is an unsubstituted or substituted group L is substituted C 1 -C 5 alkyl, the substituent group L is one or two substituents independently selected from the group: OH, C (O) OH , NH 2, pyrrolidin Alk-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorpholine-1,1-diox Generation-4-yl or NHC(O)CH(CH 3 )NHC(O)CH(CH 3 )NH;
    R’是OR 2、NR 3R 4或
    Figure PCTCN2020093248-appb-100003
    R'is OR 2 , NR 3 R 4 or
    Figure PCTCN2020093248-appb-100003
    R 2是取代基Z取代的C 1-C 5烷基，所述取代基Z是吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基或二乙醇胺基； R 2 is a C 1 -C 5 alkyl group substituted by a substituent Z, and the substituent Z is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium or diethanolamine group;
    R 3是H或C 1-C 3烷基； R 3 is H or C 1 -C 3 alkyl;
    R 4是取代基W取代的C 1-C 5烷基，所述取代基W是吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基、二乙醇胺基或乙酰基氨基； R 4 is a C 1 -C 5 alkyl group substituted by a substituent W, the substituent W is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium, diethanolamine or acetylamino;
    R 5是连接在环上任一碳上的H、OH、F、NH 2、C 1-C 3烷基氨基、COOH、C 1-C 3烷基、C 1-C 3烷氧基、环烷基氧基或杂环烷基氧基； R 5 is H, OH, F, NH 2 , C 1 -C 3 alkylamino, COOH, C 1 -C 3 alkyl, C 1 -C 3 alkoxy, cycloalkane attached to any carbon on the ring Oxy or heterocycloalkyloxy;
    Q是CH 2、O、NR 6、S、SO 2、CHR 7或化学键； Q is CH 2 , O, NR 6 , S, SO 2 , CHR 7 or chemical bond;
    R 6是H、C 1-C 3烷基、C 1-C 3烷基酰基、C 1-C 3烷基磺酰基或取代基P取代的 C 1-C 3烷基，所述取代基P是OH、COOH、NH 2或C 1-C 3烷基氨基； R 6 is H, C 1 -C 3 alkyl, C 1 -C 3 alkyl acyl, C 1 -C 3 alkylsulfonyl, or C 1 -C 3 alkyl substituted by the substituent P, the substituent P Is OH, COOH, NH 2 or C 1 -C 3 alkylamino;
    R 7是OH、NH 2、COOH、C 1-C 3烷基氨基、C 1-C 3烷氧基、环烷基氧基或杂环烷基氧基； R 7 is OH, NH 2 , COOH, C 1 -C 3 alkylamino, C 1 -C 3 alkoxy, cycloalkyloxy or heterocycloalkyloxy;
    n是0、1或2；n is 0, 1 or 2;
    m是0、1或2。m is 0, 1, or 2.
  2. 根据权利要求1所述的化合物、其药学上可接受的盐或酯或溶剂化物，其特征在于，The compound, its pharmaceutically acceptable salt or ester or solvate according to claim 1, characterized in that:
    R是H、
    Figure PCTCN2020093248-appb-100004
    或R aCO-；     R is H,
    Figure PCTCN2020093248-appb-100004
    Or R a CO-;
    R 1是H、C 1-C 5烷基或取代基Y取代的C 1-C 5烷基，所述取代基Y是OH、C(O)OH、C(O)NH 2、NH 2、NHC(O)CH 3、吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基或二乙醇胺基； R 1 is H, C 1 -C 5 alkyl substituted by substituent Y or a C 1 -C 5 alkyl, the substituent Y is OH, C (O) OH, C (O) NH 2, NH 2, NHC(O)CH 3 , pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorph Lin-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethylammonium or diethanolamino;
    R a是非取代的或取代基L取代的C 1-C 5烷基，所述取代基L是一个或两个独立地选自下列的取代基：OH、C(O)OH、NH 2、吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基或NHC(O)CH(CH 3)NHC(O)CH(CH 3)NH； R a is an unsubstituted or substituted group L is substituted C 1 -C 5 alkyl, the substituent group L is one or two substituents independently selected from the group: OH, C (O) OH , NH 2, pyrrolidin Alk-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazin-1-yl, morpholin-4-yl, thiomorpholine-1,1-diox Generation-4-yl or NHC(O)CH(CH 3 )NHC(O)CH(CH 3 )NH;
    R’是OR 2、NR 3R 4或
    Figure PCTCN2020093248-appb-100005
    R'is OR 2 , NR 3 R 4 or
    Figure PCTCN2020093248-appb-100005
    R 2是取代基Z取代的C 1-C 5烷基，所述取代基Z是吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基或二乙醇胺基； R 2 is a C 1 -C 5 alkyl group substituted by a substituent Z, and the substituent Z is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium or diethanolamine group;
    R 3是H； R 3 is H;
    R 4是取代基W取代的C 1-C 5烷基，所述取代基W是吡咯烷-1-基、哌啶-1-基、哌嗪-1-基、4-甲基-哌嗪-1-基、吗啉-4-基、硫代吗啉-1,1-二氧代-4-基、N,N-二甲基氨基、N,N-二乙基氨基、三甲基铵基、二乙醇胺基或乙酰基氨基； R 4 is a C 1 -C 5 alkyl group substituted by a substituent W, the substituent W is pyrrolidin-1-yl, piperidin-1-yl, piperazin-1-yl, 4-methyl-piperazine -1-yl, morpholin-4-yl, thiomorpholine-1,1-dioxo-4-yl, N,N-dimethylamino, N,N-diethylamino, trimethyl Ammonium, diethanolamine or acetylamino;
    R 5是连接在环上任一碳上的H、OH、F、NH 2、C 1-C 3烷基氨基或COOH； R 5 is H, OH, F, NH 2 , C 1 -C 3 alkylamino or COOH attached to any carbon on the ring;
    Q是CH 2、O、NR 6、S、SO 2或CHR 7； Q is CH 2 , O, NR 6 , S, SO 2 or CHR 7 ;
    R 6是H、C 1-C 3烷基、C 1-C 3烷基酰基、C 1-C 3烷基磺酰基或取代基P取代的C 1-C 3烷基，所述取代基P是OH、COOH、NH 2或C 1-C 3烷基氨基； R 6 is H, C 1 -C 3 alkyl, C 1 -C 3 alkyl acyl, C 1 -C 3 alkylsulfonyl, or C 1 -C 3 alkyl substituted by the substituent P, the substituent P Is OH, COOH, NH 2 or C 1 -C 3 alkylamino;
    R 7是OH、NH 2、COOH或C 1-C 3烷基氨基； R 7 is OH, NH 2 , COOH or C 1 -C 3 alkylamino;
    n是0或1；n is 0 or 1;
    m是0或1。m is 0 or 1.
  3. 根据权利要求1所述的化合物、其药学上可接受的盐或酯或溶剂化物，其特征在于，在所述式II化合物中，当取代基Z是N,N-二甲基氨基或三甲基铵基时，R不是H；当取代基W是乙酰基氨基时，R不是H。The compound according to claim 1, its pharmaceutically acceptable salt or ester or solvate, wherein in the compound of formula II, when the substituent Z is N,N-dimethylamino or trimethyl In the case of an ammonium group, R is not H; when the substituent W is an acetylamino group, R is not H.
  4. 根据权利要求1所述的化合物、其药学上可接受的盐或酯或溶剂化物，其特征在于，所述化合物或其药学上可接受的盐或酯或溶剂化物选自如下化合物：The compound, or a pharmaceutically acceptable salt or ester or solvate thereof according to claim 1, wherein the compound or a pharmaceutically acceptable salt or ester or solvate thereof is selected from the following compounds:
    Figure PCTCN2020093248-appb-100006
    Figure PCTCN2020093248-appb-100006
    Figure PCTCN2020093248-appb-100007
    Figure PCTCN2020093248-appb-100007
    Figure PCTCN2020093248-appb-100008
    Figure PCTCN2020093248-appb-100008
    Figure PCTCN2020093248-appb-100009
    Figure PCTCN2020093248-appb-100009
    Figure PCTCN2020093248-appb-100010
    Figure PCTCN2020093248-appb-100010
    Figure PCTCN2020093248-appb-100011
    Figure PCTCN2020093248-appb-100011
    Figure PCTCN2020093248-appb-100012
    Figure PCTCN2020093248-appb-100012
    Figure PCTCN2020093248-appb-100013
    Figure PCTCN2020093248-appb-100013
    Figure PCTCN2020093248-appb-100014
    Figure PCTCN2020093248-appb-100014
    Figure PCTCN2020093248-appb-100015
    Figure PCTCN2020093248-appb-100015
    Figure PCTCN2020093248-appb-100016
    Figure PCTCN2020093248-appb-100016
    Figure PCTCN2020093248-appb-100017
    Figure PCTCN2020093248-appb-100017
    Figure PCTCN2020093248-appb-100018
    Figure PCTCN2020093248-appb-100018
    Figure PCTCN2020093248-appb-100019
    Figure PCTCN2020093248-appb-100019
    Figure PCTCN2020093248-appb-100020
    Figure PCTCN2020093248-appb-100020
    Figure PCTCN2020093248-appb-100021
    Figure PCTCN2020093248-appb-100021
    Figure PCTCN2020093248-appb-100022
    Figure PCTCN2020093248-appb-100022
    Figure PCTCN2020093248-appb-100023
    Figure PCTCN2020093248-appb-100023
    Figure PCTCN2020093248-appb-100024
    Figure PCTCN2020093248-appb-100024
    Figure PCTCN2020093248-appb-100025
    Figure PCTCN2020093248-appb-100025
    Figure PCTCN2020093248-appb-100026
    Figure PCTCN2020093248-appb-100026
    Figure PCTCN2020093248-appb-100027
    Figure PCTCN2020093248-appb-100027
    Figure PCTCN2020093248-appb-100028
    Figure PCTCN2020093248-appb-100028
    Figure PCTCN2020093248-appb-100029
    Figure PCTCN2020093248-appb-100029
    Figure PCTCN2020093248-appb-100030
    Figure PCTCN2020093248-appb-100030
    Figure PCTCN2020093248-appb-100031
    Figure PCTCN2020093248-appb-100031
    Figure PCTCN2020093248-appb-100032
    Figure PCTCN2020093248-appb-100032
    Figure PCTCN2020093248-appb-100033
    Figure PCTCN2020093248-appb-100033
    Figure PCTCN2020093248-appb-100034
    Figure PCTCN2020093248-appb-100034
    Figure PCTCN2020093248-appb-100035
    Figure PCTCN2020093248-appb-100035
  5. 一种权利要求1-4任一项所述的化合物、其药学上可接受的盐或酯或溶剂化物在制备具有增强AMPK磷酸化水平活性的AMPK激动剂中的应用。A use of the compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt or ester or solvate thereof in the preparation of AMPK agonists with activity to enhance AMPK phosphorylation level.
  6. 一种权利要求1-4任一项所述的化合物、其药学上可接受的盐或酯或溶剂化物在制备预防或治疗AMPK介导的疾病的药物中的用途。A use of the compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt or ester or solvate thereof in the preparation of a medicine for preventing or treating AMPK-mediated diseases.
  7. 根据权利要6所述的用途，所述AMPK介导的疾病包括代谢性疾病、心 脑血管疾病、炎症疾病、自身免疫性疾病、器官纤维化疾病、神经退行性疾病、病原体感染所致的继发性疾病、线粒体功能障碍或紊乱疾病或肿瘤。The use according to claim 6, the AMPK-mediated diseases include metabolic diseases, cardiovascular and cerebrovascular diseases, inflammatory diseases, autoimmune diseases, organ fibrotic diseases, neurodegenerative diseases, and subsequent pathogen infections. Primary disease, mitochondrial dysfunction or disorder disease or tumor.
  8. 根据权利要6所述的用途，所述AMPK介导的疾病包括胰岛素抵抗、代谢综合征、1型或2型糖尿病、高脂血症、肥胖症、动脉粥样硬化、心肌缺血、心肌梗死、心律失常、冠心病、高血压、心衰、心肌肥大、心肌炎、糖尿病并发症、非酒精性脂肪肝、非酒精性脂肪性肝炎、酒精性脂肪肝、肝硬化、痛风、中风、脑梗死、肺炎、哮喘、慢性阻塞性肺病、慢性支气管炎、肺气肿、闭塞性细支气管炎、特发性肺纤维化、囊性纤维化肺病、过敏性鼻炎、炎性肠病、多囊肾病、多囊卵巢综合征、白塞氏病、系统性红斑狼疮、类风湿关节炎、脊椎关节炎、骨关节炎、滑膜炎、肌腱炎、血栓闭塞性脉管炎、静脉炎、间歇性跛行、瘢痕瘤、银屑病、鱼鳞癣、大疱性类天疱疮、皮炎、接触性皮炎、胰腺炎、慢性肾炎、膀胱炎、脑膜炎、胃炎、败血症、坏疽性脓皮症、葡萄膜炎、帕金森病、阿尔茨海默病、α-共核蛋白病、抑郁症、多发性硬化症、肌萎缩侧索硬化病、纤维肌痛综合症、神经痛、唐氏综合征、哈勒沃登-施帕病、亨廷顿舞蹈病、威尔逊病、肌无力、肌阵挛、运动不耐受、卡恩斯-赛尔综合征、慢性疲乏综合征、利氏综合征、线粒体肌病-脑病-高乳酸血症、中风综合征、中风样发作、杜氏肌营养不良、贝壳肌营养不良、弗立德希氏共济失调或肿瘤。The use according to claim 6, wherein the AMPK-mediated diseases include insulin resistance, metabolic syndrome, type 1 or type 2 diabetes, hyperlipidemia, obesity, atherosclerosis, myocardial ischemia, and myocardial infarction , Arrhythmia, coronary heart disease, hypertension, heart failure, myocardial hypertrophy, myocarditis, diabetic complications, non-alcoholic fatty liver, non-alcoholic steatohepatitis, alcoholic fatty liver, cirrhosis, gout, stroke, cerebral infarction, Pneumonia, asthma, chronic obstructive pulmonary disease, chronic bronchitis, emphysema, bronchiolitis obliterans, idiopathic pulmonary fibrosis, cystic fibrotic lung disease, allergic rhinitis, inflammatory bowel disease, polycystic kidney disease, multiple Cystic ovarian syndrome, Behçet's disease, systemic lupus erythematosus, rheumatoid arthritis, spondyloarthritis, osteoarthritis, synovitis, tendinitis, thromboangiitis obliterans, phlebitis, intermittent claudication, scars Tumor, psoriasis, ichthyosis, bullous pemphigoid, dermatitis, contact dermatitis, pancreatitis, chronic nephritis, cystitis, meningitis, gastritis, sepsis, pyoderma gangrenosum, uveitis, par Kinsen's disease, Alzheimer's disease, α-synuclein disease, depression, multiple sclerosis, amyotrophic lateral sclerosis, fibromyalgia syndrome, neuralgia, Down syndrome, Hallerwarden- Spar's disease, Huntington's disease, Wilson disease, myasthenia, myoclonus, exercise intolerance, Karnes-Saier syndrome, chronic fatigue syndrome, Leigh syndrome, mitochondrial myopathy-encephalopathy-high lactate Blood disease, stroke syndrome, stroke-like episodes, Duchenne muscular dystrophy, shell muscular dystrophy, Friedrich's ataxia or tumor.
  9. 一种预防或治疗AMPK介导的疾病的药物组合物，其包含如权利要求1～4任一项所述的化合物、其药学上可接受的盐或酯或溶剂化合物作为活性成分和药学上可接受的辅料。A pharmaceutical composition for preventing or treating AMPK-mediated diseases, comprising the compound according to any one of claims 1 to 4, its pharmaceutically acceptable salt or ester or solvent compound as an active ingredient and a pharmaceutically acceptable compound. Accepted excipients.
  10. [根据细则91更正 09.06.2020]　
    根据权利要求9所述的药物组合物，其特征在于，所述药物组合物为胶囊剂、散剂、片剂、颗粒剂、丸剂、注射剂、糖浆剂、口服液、吸入剂、软膏剂、栓剂或贴剂。     [Corrected according to Rule 91 09.06.2020]
    The pharmaceutical composition according to claim 9, wherein the pharmaceutical composition is a capsule, powder, tablet, granule, pill, injection, syrup, oral liquid, inhalant, ointment, suppository or Patches.
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