WO2020259205A1 - 齐墩果酸和δ-齐墩果酸的衍生物及其医药用途 - Google Patents
齐墩果酸和δ-齐墩果酸的衍生物及其医药用途 Download PDFInfo
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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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- 0 CC(C)(CC[C@@]1(CC2)C3(*C3)NCCCN3CCOCC3)CC1=C(CC1)[C@]2(C)[C@](C)(CC2)[C@]1[C@@](C)(CC1)[C@]2C(C)(C)[C@]1OC(c(cccc1)c1C(*)=O)=O Chemical compound CC(C)(CC[C@@]1(CC2)C3(*C3)NCCCN3CCOCC3)CC1=C(CC1)[C@]2(C)[C@](C)(CC2)[C@]1[C@@](C)(CC1)[C@]2C(C)(C)[C@]1OC(c(cccc1)c1C(*)=O)=O 0.000 description 3
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- CQKIESZHYZHCLH-ZHTUXDAXSA-O CC(C)(CC1)CC2C3=CC[C@H]([C@@](C)(CC4)[C@@H](CC5)C(C)(C)C4OC(c(cccc4)c4C(OCCNC(C)=O)=O)=O)[C@]5(C)[C@]3(C)CC[C@@]12C(OCC[N+](C)(C)C)=O Chemical compound CC(C)(CC1)CC2C3=CC[C@H]([C@@](C)(CC4)[C@@H](CC5)C(C)(C)C4OC(c(cccc4)c4C(OCCNC(C)=O)=O)=O)[C@]5(C)[C@]3(C)CC[C@@]12C(OCC[N+](C)(C)C)=O CQKIESZHYZHCLH-ZHTUXDAXSA-O 0.000 description 1
- ZWWODWNIOFZGBL-AOBZOHDZSA-N CC(C)(CC1)C[C@H]2C3=CC[C@H]([C@@](C)(CC4)[C@@H](CC5)C(C)(C)[C@H]4O)[C@]5(C)[C@]3(C)CC[C@@]12C(CCC(CC1)C[C@@H]1O)=O Chemical compound CC(C)(CC1)C[C@H]2C3=CC[C@H]([C@@](C)(CC4)[C@@H](CC5)C(C)(C)[C@H]4O)[C@]5(C)[C@]3(C)CC[C@@]12C(CCC(CC1)C[C@@H]1O)=O ZWWODWNIOFZGBL-AOBZOHDZSA-N 0.000 description 1
- ZAGMIOFMYUJFBA-JNNZJYSRSA-N CC(C)(CC1)C[C@H]2C3=CC[C@H]([C@@](C)(CC4)[C@@H](CC5)C(C)(C)[C@H]4OC(c(cccc4)c4C(O)=O)=O)[C@]5(C)[C@]3(C)CC[C@@]12C(NCCCN(CC1)CCS1(=O)=O)=O Chemical compound CC(C)(CC1)C[C@H]2C3=CC[C@H]([C@@](C)(CC4)[C@@H](CC5)C(C)(C)[C@H]4OC(c(cccc4)c4C(O)=O)=O)[C@]5(C)[C@]3(C)CC[C@@]12C(NCCCN(CC1)CCS1(=O)=O)=O ZAGMIOFMYUJFBA-JNNZJYSRSA-N 0.000 description 1
- PXFPPBOZTBOWGR-AFZFOALMSA-N CC(C)(CC1)C[C@H]2C3=CC[C@H]([C@@](C)(CC4)[C@@H](CC5)C(C)(C)[C@H]4OC(c4nc(F)ccc4)=O)[C@]5(C)[C@]3(C)CC[C@@]12C(CCC(C1)CC1OC)=O Chemical compound CC(C)(CC1)C[C@H]2C3=CC[C@H]([C@@](C)(CC4)[C@@H](CC5)C(C)(C)[C@H]4OC(c4nc(F)ccc4)=O)[C@]5(C)[C@]3(C)CC[C@@]12C(CCC(C1)CC1OC)=O PXFPPBOZTBOWGR-AFZFOALMSA-N 0.000 description 1
- DWUMPZIKXSKCJF-FLLZMTHVSA-N CC(C)(CC[C@@]1(CC2)C(N3CCOCC3)=O)CC1=C(CC1)[C@]2(C)[C@](C)(CC2)[C@H]1[C@@](C)(CC1)[C@@H]2C(C)(C)C1OC(c(cccc1)c1C(O)=O)=O Chemical compound CC(C)(CC[C@@]1(CC2)C(N3CCOCC3)=O)CC1=C(CC1)[C@]2(C)[C@](C)(CC2)[C@H]1[C@@](C)(CC1)[C@@H]2C(C)(C)C1OC(c(cccc1)c1C(O)=O)=O DWUMPZIKXSKCJF-FLLZMTHVSA-N 0.000 description 1
- UTBNLEVTWWNKDX-ZAZZEFMQSA-N CC(C)(CC[C@@]1(CC2)C(NCCN3CCN(C)CC3)=O)CC1=C(CC1)[C@]2(C)C(C)(CC2)[C@H]1[C@@](C)(CC1)[C@@H]2C(C)(C)[C@H]1OC(c(cccc1)c1C(O)=O)=O Chemical compound CC(C)(CC[C@@]1(CC2)C(NCCN3CCN(C)CC3)=O)CC1=C(CC1)[C@]2(C)C(C)(CC2)[C@H]1[C@@](C)(CC1)[C@@H]2C(C)(C)[C@H]1OC(c(cccc1)c1C(O)=O)=O UTBNLEVTWWNKDX-ZAZZEFMQSA-N 0.000 description 1
- DNPNYIXBTPQDIS-FJZZLOJVSA-N CC(C)OC(c(cccc1)c1C(O[C@@H](CC1)C(C)(C)[C@H](CC2)[C@@]1(C)[C@@H]1[C@]2(C)[C@](C)(CC[C@@]2([C@H]3CC(C)(C)CC2)C(NCCN2CCCCC2)=O)C3=CC1)=O)=O Chemical compound CC(C)OC(c(cccc1)c1C(O[C@@H](CC1)C(C)(C)[C@H](CC2)[C@@]1(C)[C@@H]1[C@]2(C)[C@](C)(CC[C@@]2([C@H]3CC(C)(C)CC2)C(NCCN2CCCCC2)=O)C3=CC1)=O)=O DNPNYIXBTPQDIS-FJZZLOJVSA-N 0.000 description 1
- QEGNUYASOUJEHD-UHFFFAOYSA-N CC1(C)CCCCC1 Chemical compound CC1(C)CCCCC1 QEGNUYASOUJEHD-UHFFFAOYSA-N 0.000 description 1
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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
本发明公开了一种五环三萜类新型AMPK激动剂齐墩果酸和δ-齐墩果酸的衍生物及其医药用途，具体如式I或式II所示的化合物、其药学上可接受的盐或酯或溶剂化物，它们可用制备具有增强AMPK磷酸化水平活性的AMPK激动剂以及用于制备预防或治疗AMPK介导的疾病的药物。本发明的新型五环三萜类化合物具有显著的AMPK激动活性，且其活性显著优于公认的AMPK激动剂AICAR，同时具有更好的口服生物利用度等药代动力学性质和非常好的安全性。
Description
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。
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)。
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。
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)。
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。
发明内容
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。
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。
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：
其中，
R是H、
或R
aCO-；
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
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’是OR
2、NR
3R
4或
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或C
1-C
3烷基；
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
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；
Q是CH
2、O、NR
6、S、SO
2、CHR
7或化学键；
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
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；
n是0、1或2；
m是0、1或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，
R是H、
或R
aCO-；
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
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’是OR
2、NR
3R
4或
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
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
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；
Q是CH
2、O、NR
6、S、SO
2或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
7是OH、NH
2、COOH或C
1-C
3烷基氨基；
n是0或1；
m是0或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。
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：
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。
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。
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。
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。
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。
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。
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。
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。
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。
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。
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。
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。
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。
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。
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。
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。
有益效果：与现有技术相比，本发明具有如下优点：
(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。
(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。
(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。
(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。
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)。
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。
实施例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)
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％)。
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。
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).
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％)。
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％)。
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]
+。
实施例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)
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％)。
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％)。
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％)。
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％)。
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]
-。
实施例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)
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]
+。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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％)。
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％)。
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％)。
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％)。
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％)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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％)。
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％)。
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]
+。
实施例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)
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]
+。
实施例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)
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％)。
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]
+。
实施例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)
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).
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
-。
实施例26
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-吗啉(化合物A-84)
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]
+。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例31
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-哌啶(化合物A-104)
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]
+。
实施例32
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-哌嗪(化合物A-86)
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]
+。
实施例33
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-四氢吡咯(化合物A-88)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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。
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]
-。
实施例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)
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]
+。
实施例38
N-(3β-羟基-齐墩果烷-13(18)-烯-28-酰基)-吖丁啶(化合物A-95)
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]
+。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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。
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]
+。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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％)。
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。
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]
-。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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％)。
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％)。
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]
-。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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％)。
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]
+。
实施例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)
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]
+。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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％)。
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％)。
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％)。
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]
+。
实施例74
N-(3β-羟基-齐墩果烷-12-烯-28-酰胺)-吗啉(化合物B-2)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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。
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]
-。
实施例87
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-甲基-哌嗪(化合物B-17)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例93
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-哌嗪(化合物B-25)
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。
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]
+。
实施例94
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-四氢吡咯(化合物B-26)
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]
+。
实施例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)
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。
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例99
3β-羟基-齐墩果烷-12-烯-28-酸2-(1-哌啶基)乙酯(化合物B-31)
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％)。
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]
+。
实施例100
3β-羟基-齐墩果烷-12-烯-28-酸2-(4-吗啉基)乙酯(化合物B-32)
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]
+。
实施例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)
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]
+。
实施例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)
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。
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]
-。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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％)。
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％)。
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％)。
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％)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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％)。
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例141
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-吖丁啶(化合物B-106)
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]
+。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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。
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]
-。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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％)。
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]
+。
实施例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)
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]
+。
实施例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)
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％)。
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％)。
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％)。
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]
+。
实施例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)
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。
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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％)。
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]
+。
实施例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)
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]
+。
实施例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)
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％)。
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％)。
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％)。
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％)。
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例172
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-苄基-哌嗪(化合物B-133)
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]
+。
实施例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)
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]
+。
实施例174
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-乙基-哌嗪(化合物B-135)
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]
+。
实施例175
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-氧代-哌啶(化合物B-136)
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]
+。
实施例176
N-(3β-羟基-齐墩果烷-12-烯-28-酰基)-4-羟基-哌啶(化合物B-137)
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]
+。
实施例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)
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]
-。
实施例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)
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]
-。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例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)
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]
+。
实施例184
化合物对Huh-7细胞AMPK激动作用的活性评价
采用Western Blot方法检测化合物对Huh-7细胞AMPK的激动活性。
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。
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)。
3、Western Blot实验：检测化合物对Huh-7细胞AMPK磷酸化水平的影响。
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：
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。
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。
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。
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。
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。
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。
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)
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。
实施例185
化合物对Huh-7细胞AMPK下游信号通路的影响
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。
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。
抗体：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：
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。
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。
实施例186
片剂
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。
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。
Claims (10)
- 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：
R是H、或R aCO-；
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 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’是OR 2、NR 3R 4或
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或C 1-C 3烷基；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 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；Q是CH 2、O、NR 6、S、SO 2、CHR 7或化学键；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 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；n是0、1或2；m是0、1或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，R是H、或R aCO-；
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 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’是OR 2、NR 3R 4或
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 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 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；Q是CH 2、O、NR 6、S、SO 2或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 7是OH、NH 2、COOH或C 1-C 3烷基氨基；n是0或1；m是0或1。 - 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。
- 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：
- 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。
- 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。
- 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。
- 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。
- 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。
- [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。
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| CN115806580A (zh) * | 2021-09-14 | 2023-03-17 | 中国医学科学院药物研究所 | 12-o-脂肪酸酯齐墩果酸类化合物及其制备方法和用途 |
| CN115873061A (zh) * | 2021-09-28 | 2023-03-31 | 中国医学科学院药物研究所 | 齐墩果酸12-o-取代乙酸酯化合物及其制备方法及用途 |
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| CN112110978B (zh) * | 2020-10-13 | 2023-02-28 | 中国药科大学 | δ-齐墩果酸皂苷类化合物及其医药用途 |
| CN115873060A (zh) * | 2021-09-28 | 2023-03-31 | 中国医学科学院药物研究所 | 12-o-苯甲酸酯齐墩果酸类化合物及其制备与用途 |
| CN117343122B (zh) * | 2023-09-08 | 2025-09-02 | 安徽医科大学 | 一种治疗炎症免疫性疾病的齐墩果酸酰胺衍生物、其制备方法及其用途 |
| CN117379446B (zh) * | 2023-12-12 | 2024-03-15 | 吉林大学 | 齐墩果酸-28-O-β-D-吡喃葡萄糖苷在制备抗结肠炎药物中的应用 |
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| CN115806580A (zh) * | 2021-09-14 | 2023-03-17 | 中国医学科学院药物研究所 | 12-o-脂肪酸酯齐墩果酸类化合物及其制备方法和用途 |
| CN115873061A (zh) * | 2021-09-28 | 2023-03-31 | 中国医学科学院药物研究所 | 齐墩果酸12-o-取代乙酸酯化合物及其制备方法及用途 |
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