CN117105895A - Tubesan sesterterpene colquhouund D derivative and preparation method and application thereof - Google Patents
Tubesan sesterterpene colquhouund D derivative and preparation method and application thereof Download PDFInfo
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- CN117105895A CN117105895A CN202311043196.8A CN202311043196A CN117105895A CN 117105895 A CN117105895 A CN 117105895A CN 202311043196 A CN202311043196 A CN 202311043196A CN 117105895 A CN117105895 A CN 117105895A
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- derivative
- colquhounod
- reaction
- organic solvent
- dissolving
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- 238000002360 preparation method Methods 0.000 title claims abstract description 112
- 229930002368 sesterterpene Natural products 0.000 title claims abstract description 14
- 150000002653 sesterterpene derivatives Chemical class 0.000 title description 3
- 239000003814 drug Substances 0.000 claims abstract description 28
- -1 dioxane sesterterpene Chemical class 0.000 claims abstract description 15
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 13
- 230000003110 anti-inflammatory effect Effects 0.000 claims abstract description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 106
- 238000006243 chemical reaction Methods 0.000 claims description 105
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 82
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 81
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 81
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 77
- 239000003960 organic solvent Substances 0.000 claims description 66
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 64
- 150000001875 compounds Chemical class 0.000 claims description 59
- 238000004809 thin layer chromatography Methods 0.000 claims description 58
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 52
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 46
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 46
- 238000003756 stirring Methods 0.000 claims description 42
- 239000012295 chemical reaction liquid Substances 0.000 claims description 35
- 239000012298 atmosphere Substances 0.000 claims description 34
- 239000011261 inert gas Substances 0.000 claims description 34
- 239000000243 solution Substances 0.000 claims description 33
- 238000010438 heat treatment Methods 0.000 claims description 29
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 28
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 24
- 229960000549 4-dimethylaminophenol Drugs 0.000 claims description 23
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 17
- 238000001035 drying Methods 0.000 claims description 17
- 239000003153 chemical reaction reagent Substances 0.000 claims description 16
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 15
- 239000000706 filtrate Substances 0.000 claims description 15
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 claims description 14
- 150000001412 amines Chemical class 0.000 claims description 13
- FMXSYRBHGUMFBA-UHFFFAOYSA-N 6-amino-3-azaniumylidene-9-[2-carboxy-4-[6-[4-[4-[4-[4-[3-carboxy-6-[4-(trifluoromethyl)phenyl]naphthalen-1-yl]phenyl]piperidin-1-yl]butyl]triazol-1-yl]hexylcarbamoyl]phenyl]-5-sulfoxanthene-4-sulfonate Chemical compound Nc1ccc2c(-c3ccc(cc3C(O)=O)C(=O)NCCCCCCn3cc(CCCCN4CCC(CC4)c4ccc(cc4)-c4cc(cc5cc(ccc45)-c4ccc(cc4)C(F)(F)F)C(O)=O)nn3)c3ccc(=[NH2+])c(c3oc2c1S(O)(=O)=O)S([O-])(=O)=O FMXSYRBHGUMFBA-UHFFFAOYSA-N 0.000 claims description 12
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims description 12
- PAYRUJLWNCNPSJ-UHFFFAOYSA-N Aniline Chemical compound NC1=CC=CC=C1 PAYRUJLWNCNPSJ-UHFFFAOYSA-N 0.000 claims description 12
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 12
- 125000001424 substituent group Chemical group 0.000 claims description 12
- 238000001914 filtration Methods 0.000 claims description 11
- 239000003018 immunosuppressive agent Substances 0.000 claims description 11
- 238000010791 quenching Methods 0.000 claims description 11
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 10
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 9
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 9
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims description 9
- 230000000171 quenching effect Effects 0.000 claims description 9
- 239000007821 HATU Substances 0.000 claims description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- PCLIMKBDDGJMGD-UHFFFAOYSA-N N-bromosuccinimide Chemical compound BrN1C(=O)CCC1=O PCLIMKBDDGJMGD-UHFFFAOYSA-N 0.000 claims description 8
- SORGEQQSQGNZFI-UHFFFAOYSA-N [azido(phenoxy)phosphoryl]oxybenzene Chemical compound C=1C=CC=CC=1OP(=O)(N=[N+]=[N-])OC1=CC=CC=C1 SORGEQQSQGNZFI-UHFFFAOYSA-N 0.000 claims description 8
- 238000007865 diluting Methods 0.000 claims description 8
- JQWHASGSAFIOCM-UHFFFAOYSA-M sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 claims description 8
- PVOAHINGSUIXLS-UHFFFAOYSA-N 1-Methylpiperazine Chemical compound CN1CCNCC1 PVOAHINGSUIXLS-UHFFFAOYSA-N 0.000 claims description 7
- 150000001540 azides Chemical class 0.000 claims description 7
- SIPUZPBQZHNSDW-UHFFFAOYSA-N bis(2-methylpropyl)aluminum Chemical compound CC(C)C[Al]CC(C)C SIPUZPBQZHNSDW-UHFFFAOYSA-N 0.000 claims description 7
- LJCNRYVRMXRIQR-OLXYHTOASA-L potassium sodium L-tartrate Chemical compound [Na+].[K+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O LJCNRYVRMXRIQR-OLXYHTOASA-L 0.000 claims description 7
- JKANAVGODYYCQF-UHFFFAOYSA-N prop-2-yn-1-amine Chemical compound NCC#C JKANAVGODYYCQF-UHFFFAOYSA-N 0.000 claims description 7
- PPASLZSBLFJQEF-RKJRWTFHSA-M sodium ascorbate Substances [Na+].OC[C@@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RKJRWTFHSA-M 0.000 claims description 7
- 235000010378 sodium ascorbate Nutrition 0.000 claims description 7
- 229960005055 sodium ascorbate Drugs 0.000 claims description 7
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 7
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 7
- 229910000104 sodium hydride Inorganic materials 0.000 claims description 7
- PPASLZSBLFJQEF-RXSVEWSESA-M sodium-L-ascorbate Chemical compound [Na+].OC[C@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RXSVEWSESA-M 0.000 claims description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 6
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 claims description 6
- 125000001494 2-propynyl group Chemical group [H]C#CC([H])([H])* 0.000 claims description 6
- 206010009900 Colitis ulcerative Diseases 0.000 claims description 6
- KEAYESYHFKHZAL-UHFFFAOYSA-N Sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 claims description 6
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 6
- 201000006704 Ulcerative Colitis Diseases 0.000 claims description 6
- UENWRTRMUIOCKN-UHFFFAOYSA-N benzyl thiol Chemical compound SCC1=CC=CC=C1 UENWRTRMUIOCKN-UHFFFAOYSA-N 0.000 claims description 6
- JZCCFEFSEZPSOG-UHFFFAOYSA-L copper(II) sulfate pentahydrate Chemical compound O.O.O.O.O.[Cu+2].[O-]S([O-])(=O)=O JZCCFEFSEZPSOG-UHFFFAOYSA-L 0.000 claims description 6
- RAFNCPHFRHZCPS-UHFFFAOYSA-N di(imidazol-1-yl)methanethione Chemical compound C1=CN=CN1C(=S)N1C=CN=C1 RAFNCPHFRHZCPS-UHFFFAOYSA-N 0.000 claims description 6
- ZBCBWPMODOFKDW-UHFFFAOYSA-N diethanolamine Chemical compound OCCNCCO ZBCBWPMODOFKDW-UHFFFAOYSA-N 0.000 claims description 6
- 239000001257 hydrogen Substances 0.000 claims description 6
- 229910052739 hydrogen Inorganic materials 0.000 claims description 6
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 6
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 claims description 6
- 229940074439 potassium sodium tartrate Drugs 0.000 claims description 6
- 239000012312 sodium hydride Substances 0.000 claims description 6
- 235000011006 sodium potassium tartrate Nutrition 0.000 claims description 6
- GZFGOTFRPZRKDS-UHFFFAOYSA-N 4-bromophenol Chemical compound OC1=CC=C(Br)C=C1 GZFGOTFRPZRKDS-UHFFFAOYSA-N 0.000 claims description 5
- 229940124599 anti-inflammatory drug Drugs 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 239000008187 granular material Substances 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- NHQDETIJWKXCTC-UHFFFAOYSA-N 3-chloroperbenzoic acid Chemical compound OOC(=O)C1=CC=CC(Cl)=C1 NHQDETIJWKXCTC-UHFFFAOYSA-N 0.000 claims description 4
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- YBCAZPLXEGKKFM-UHFFFAOYSA-K ruthenium(iii) chloride Chemical compound [Cl-].[Cl-].[Cl-].[Ru+3] YBCAZPLXEGKKFM-UHFFFAOYSA-K 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- NOGFHTGYPKWWRX-UHFFFAOYSA-N 2,2,6,6-tetramethyloxan-4-one Chemical compound CC1(C)CC(=O)CC(C)(C)O1 NOGFHTGYPKWWRX-UHFFFAOYSA-N 0.000 claims description 3
- GQHTUMJGOHRCHB-UHFFFAOYSA-N 2,3,4,6,7,8,9,10-octahydropyrimido[1,2-a]azepine Chemical compound C1CCCCN2CCCN=C21 GQHTUMJGOHRCHB-UHFFFAOYSA-N 0.000 claims description 3
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 claims description 3
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 claims description 3
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 3
- 229960002685 biotin Drugs 0.000 claims description 3
- 235000020958 biotin Nutrition 0.000 claims description 3
- 239000011616 biotin Substances 0.000 claims description 3
- 239000002775 capsule Substances 0.000 claims description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 3
- 125000001559 cyclopropyl group Chemical class [H]C1([H])C([H])([H])C1([H])* 0.000 claims description 3
- IKGLACJFEHSFNN-UHFFFAOYSA-N hydron;triethylazanium;trifluoride Chemical compound F.F.F.CCN(CC)CC IKGLACJFEHSFNN-UHFFFAOYSA-N 0.000 claims description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 3
- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 claims description 3
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 claims description 3
- 150000002540 isothiocyanates Chemical class 0.000 claims description 3
- 125000000740 n-pentyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 claims description 3
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 claims description 3
- CYQAYERJWZKYML-UHFFFAOYSA-N phosphorus pentasulfide Chemical compound S1P(S2)(=S)SP3(=S)SP1(=S)SP2(=S)S3 CYQAYERJWZKYML-UHFFFAOYSA-N 0.000 claims description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 3
- WETWJCDKMRHUPV-UHFFFAOYSA-N acetyl chloride Chemical compound CC(Cl)=O WETWJCDKMRHUPV-UHFFFAOYSA-N 0.000 claims description 2
- 239000012346 acetyl chloride Substances 0.000 claims description 2
- PASDCCFISLVPSO-UHFFFAOYSA-N benzoyl chloride Chemical compound ClC(=O)C1=CC=CC=C1 PASDCCFISLVPSO-UHFFFAOYSA-N 0.000 claims description 2
- 239000012267 brine Substances 0.000 claims description 2
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 claims description 2
- 239000002260 anti-inflammatory agent Substances 0.000 claims 2
- 229940124589 immunosuppressive drug Drugs 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- NNQDMQVWOWCVEM-UHFFFAOYSA-N 1-bromoprop-1-ene Chemical compound CC=CBr NNQDMQVWOWCVEM-UHFFFAOYSA-N 0.000 claims 1
- XXFUZSHTIOFGNV-UHFFFAOYSA-N 1-bromoprop-1-yne Chemical compound CC#CBr XXFUZSHTIOFGNV-UHFFFAOYSA-N 0.000 claims 1
- BLXSFCHWMBESKV-UHFFFAOYSA-N 1-iodopentane Chemical class CCCCCI BLXSFCHWMBESKV-UHFFFAOYSA-N 0.000 claims 1
- AGEZXYOZHKGVCM-UHFFFAOYSA-N benzyl bromide Chemical compound BrCC1=CC=CC=C1 AGEZXYOZHKGVCM-UHFFFAOYSA-N 0.000 claims 1
- KMGBZBJJOKUPIA-UHFFFAOYSA-N butyl iodide Chemical class CCCCI KMGBZBJJOKUPIA-UHFFFAOYSA-N 0.000 claims 1
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical class IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 claims 1
- DUCPEQVHPGJMED-UHFFFAOYSA-N isothiocyanic acid;pyridine Chemical class N=C=S.C1=CC=NC=C1 DUCPEQVHPGJMED-UHFFFAOYSA-N 0.000 claims 1
- PVWOIHVRPOBWPI-UHFFFAOYSA-N n-propyl iodide Chemical class CCCI PVWOIHVRPOBWPI-UHFFFAOYSA-N 0.000 claims 1
- QKFJKGMPGYROCL-UHFFFAOYSA-N phenyl isothiocyanate Chemical class S=C=NC1=CC=CC=C1 QKFJKGMPGYROCL-UHFFFAOYSA-N 0.000 claims 1
- 229940098458 powder spray Drugs 0.000 claims 1
- 239000003826 tablet Substances 0.000 claims 1
- 230000001506 immunosuppresive effect Effects 0.000 abstract description 21
- 229940079593 drug Drugs 0.000 abstract description 19
- 206010062016 Immunosuppression Diseases 0.000 abstract description 6
- 239000004480 active ingredient Substances 0.000 abstract description 4
- 238000009776 industrial production Methods 0.000 abstract description 2
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- CSCPPACGZOOCGX-MICDWDOJSA-N 1-deuteriopropan-2-one Chemical compound [2H]CC(C)=O CSCPPACGZOOCGX-MICDWDOJSA-N 0.000 description 89
- 239000011734 sodium Substances 0.000 description 84
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 46
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- 230000002829 reductive effect Effects 0.000 description 24
- 229910052786 argon Inorganic materials 0.000 description 23
- 239000012074 organic phase Substances 0.000 description 22
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 21
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- 239000007858 starting material Substances 0.000 description 19
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 18
- 238000004440 column chromatography Methods 0.000 description 18
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- TVZRAEYQIKYCPH-UHFFFAOYSA-N 3-(trimethylsilyl)propane-1-sulfonic acid Chemical compound C[Si](C)(C)CCCS(O)(=O)=O TVZRAEYQIKYCPH-UHFFFAOYSA-N 0.000 description 12
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D307/93—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems condensed with a ring other than six-membered
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
- C07D491/08—Bridged systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
- C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
- C07D495/10—Spiro-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/07—Optical isomers
Abstract
The invention discloses a colquhouund D derivative of a colquhouund with a general formula (I), a preparation method thereof, a pharmaceutical composition taking the colquhouund D derivative as an active ingredient and application thereof in pharmacy. Belongs to the technical field of medicines. The method for preparing the colquhouund D derivative is easy to obtain raw materials, easy to operate, high in yield and suitable for industrial production. The pharmaceutical composition taking the dioxane sesterterpene colquhounod D derivative as an active ingredient provides a novel medicament with better medicinal effect for novel anti-inflammatory medicaments and immunosuppression medicaments.
Description
Technical Field
The invention relates to the technical field of pharmaceutical chemistry, in particular to a derivative prepared from a natural product of sesterterpene colquhounod D from fireflower Colquhounia coccinea wall. Var. Mollis (Schlecht.) Prain, a preparation method and application thereof.
Background
Natural drugs are an important component of drugs, and sources of natural drugs include plants, animals, minerals and microorganisms, and are mainly plants, and are various. It was counted that 25% of the new chemical entity drugs approved from 34 years 1981-2014 were natural products or derivatives thereof, 26% were artificially synthesized natural products and natural product analogs. The biological products and vaccine are removed, and the natural products or derivatives thereof account for 32% in 1211 new chemical entity small molecule drugs, and the artificially synthesized natural products and natural product analogues account for 32%. It follows that natural products are of great importance in the development of new drugs. The case of directly preparing medicines from natural products or preparing medicines after structure optimization is also not enumerated. In 1820, the French pharmacist Caventou and Pelletier reported the isolation of antimalarial Quinine (Quinine) from Cinchona (Cinchona). Sesquiterpene lactone compound artemisinin (argon temp) and its analogues isolated from Artemisia annua (argon temp) are effective components for resisting malaria. The anticancer drug Paclitaxel (Paclitaxel) is diterpenoid compound separated from bark of Taxus brevifolia (Taxus brevensis).
Autoimmune diseases are affected by a variety of factors, such as genetics, environment, hormones, etc., and there is no single theory or mechanism that can fully explain all disease features or pathogenesis. Common autoimmune diseases include rheumatoid arthritis (rheumatoid arthritis, RA), multiple sclerosis (multiple sclerosis, MS), systemic lupus erythematosus (systemic lupus erythematosus, SLE), and the like. Autoimmune diseases often have a long course, not only bringing physiological damage to the patient, but also psychological affliction. In the case of rheumatoid arthritis, the patient gradually develops joint injury with the progress of the disease, and finally causes joint deformity and loss of function. In China, the disability rates of rheumatoid arthritis patients in 1-5 years, 5-10 years, 10-15 years and more than or equal to 15 years are respectively 18.6%, 43.5%, 48.1% and 61.3%, and the occurrence rate of disability and limited functions is obviously increased along with the extension of the disease course. Rheumatoid arthritis not only causes the decline of physical functions, life quality and social engagement of patients, but also brings great economic burden to families and society of patients.
The small molecule immunosuppressant is a preparation with the function of inhibiting organism immunity, and is mainly used for controlling organ transplant rejection, and treating various autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis and the like. Currently, clinically commonly used small molecule immunosuppressants mainly include Tacrolimus (Tacrolimus), cyclophosphamide (CTX), and glucocorticoids (glucomoricids), which have remarkable therapeutic effects, but also have many adverse effects. For example, glucocorticoids cause adverse effects such as osteoporosis, diabetes, glaucoma, etc.; CTX can lead to bone marrow suppression, nausea, vomiting, hair loss, etc. Tacrolimus has better selectivity, but long-term administration results in increased hyperglycemia, neurotoxicity, nephrotoxicity, and tumor incidence. In addition, tofacitinib, fingolimod and other novel small molecule immunosuppressants are mainly used for treating autoimmune diseases. The novel small molecule immunosuppressants have definite molecular targets and specific target protein expression, and compared with the traditional immunosuppressants, the novel small molecule immunosuppressants have better therapeutic effect and smaller toxic effect, and do not cause systemic adverse reactions. Therefore, searching for immunosuppressants with definite and novel action targets, good activity and few side effects would be a long-term goal for future immunosuppressant development.
At present, 37 novel skeleton-labdane sesterterpenes with defense function have been reported to be isolated from the Labiatae family plant, fireflower Colquhounia coccinea wall. Var. Mollis (Schlecht.) Prain, and colquhound D is the main ingredient thereof. When the above-mentioned firefly alkane sesterterpenes were subjected to activity screening, they were found to exhibit immunosuppressive activity in large numbers. Therefore, the colquhounod D is subjected to structural modification, and through activity screening, evaluation and structural optimization, a lead molecule with stronger immunosuppressive activity is found to have potential, and a material basis can be provided for safer and more effective immunosuppressant research and development.
Disclosure of Invention
In view of the above, the present invention aims to provide a compound precursor-a colquhouund D derivative with immunosuppressive activity, a pharmaceutical composition containing the same as an active ingredient, a preparation method thereof, and applications thereof in preparing medicines for treating inflammation and immunosuppression and medicines for treating ulcerative colitis.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
a colquhouund D derivative of a labdane sesterterpene shown in the following structural formula (I),
In the general formula (I), R 1 Substituents include, but are not limited to, hydrogen, methyl, n-propyl, n-butyl, n-pentyl, allyl, propargyl, benzoyl, acetyl,
R 2 Substituents include, but are not limited to, carboxyl, hydroxymethyl,
R 3 And R is R 4 Substituents include, but are not limited to, substituted or unsubstituted cyclopropane, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, hydrogen atom;
R 5 substituents include, but are not limited to, hydrogen, methyl, allyl, propargyl, benzyl, biotin;
R 6 substituents include, but are not limited to, hydroxy.
According to the derivative of the colquhouund D of the firefly alkane sesterterpene shown in the general formula (I), the derivative is any one of the following compounds:
the invention also discloses a preparation method of the colquhounod D derivative 1-21, which comprises the following steps:
(1) Dissolving a compound colquhounod D shown in a structural formula (II) in an organic solvent;
(2) Adding 1.5M diisobutyl aluminum hydride in inert gas atmosphere, heating and stirring to react until TLC shows that the reaction is complete, quenching the reaction liquid with potassium sodium tartrate, extracting, drying, concentrating in vacuum, and purifying to obtain a colquhounod D derivative 1-4;
(3) Dissolving the colquhounod D derivative 1 in an organic solvent;
(4) Under inert gas atmosphere, adding an acylating reagent, DMAP and triethylamine, heating and stirring until TLC shows that the reaction is complete, and concentrating and purifying the reaction liquid in vacuum to obtain the colquhounodD derivative 5-6 in claim 2.
(5) Dissolving the colquhounod D derivative 1 in an organic solvent;
(6) And adding N, N-thiocarbonyldiimidazole and DMAP under inert gas atmosphere, heating and stirring until TLC shows that the reaction is complete, and concentrating and purifying the reaction liquid in vacuum to obtain the colquhounod D derivative 8-9.
(7) Dissolving the colquhounod D derivative 1 in an organic solvent;
(8) Adding sodium hydride, different substituted isothiocyanates or halogenated hydrocarbons in inert gas atmosphere, heating and stirring to react until TLC shows complete reaction, filtering the reaction liquid, collecting filtrate, concentrating in vacuum, and purifying to obtain the colquhounod D derivative 10-21.
(9) Dissolving the colquhounod D derivative 6 in an organic solvent;
(10) And adding phosphorus pentasulfide and sodium carbonate under inert gas atmosphere, heating and stirring until TLC shows that the reaction is complete, and concentrating and purifying the reaction liquid in vacuum to obtain the colquhounod D derivative 7.
Preferably, in the above preparation method of the colquhouoid D derivative 1-21, the organic solvent in the step (1), the step (3), the step (5), the step (7) and the step (9) is one or more selected from N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine and ethanol, the step (1), the step (7) and the step (9) are more preferably tetrahydrofuran, the step (3) is more preferably dichloromethane, and the step (5) is more preferably toluene.
Preferably, in the above preparation method of the colquhounod D derivative 1-21, the acylating agent in the step (4) is selected from one of acetic anhydride, benzoic anhydride, acetyl chloride and benzoyl chloride, and more preferably acetic anhydride or benzoic anhydride.
Preferably, in the preparation method of the colquhounod D derivative 1-21, the heating temperature in the step (1) is 0-60 ℃, more preferably the heating temperature of the derivative 1-3 is 60 ℃, and the heating temperature of the derivative 4 is 0 ℃; the heating temperature in the step (6) is 60-80 ℃, and further preferably 80 ℃; the heating temperature in the step (8) is 25-60 ℃, more preferably the heating temperature of the derivative 10-13 is 25 ℃, the heating temperature of the derivative 14-19 is 45 ℃, and the heating temperature of the derivative 20-21 is 60 ℃; the heating temperature in step (10) is 40℃to 80℃and more preferably 80 ℃.
Preferably, in the preparation method of the colquhounod D derivative 1-21, the molar ratio of 1.5M DIBAl-H to colquhounod D is 20:1.
Preferably, in the preparation method of the colquhounod D derivative 1-21, the molar ratio of the acylating agent, DMAP, triethylamine and the colquhounod D derivative 1 is 46.2:1:31.6:1.
Preferably, in the preparation method of the colquhounod D derivative 1-21, the molar ratio of the N, N-thiocarbonyldiimidazole, the DMAP and the colquhounod D derivative 1 is 2:3:1.
Preferably, in the preparation method of the colquhounod D derivative 1-21, the molar ratio of the sodium hydride, the isothiocyanate or the halogenated hydrocarbon to the colquhounod D derivative 1 is 30:35:1.
Preferably, in the preparation method of the colquhounod D derivative 1-21, the molar ratio of the phosphorus pentasulfide, the sodium carbonate and the colquhounod D derivative 6 is 1:1:1.
The invention also discloses a preparation method of the colquhounod D derivative 22-38, which comprises the following steps:
(1) Dissolving a compound colquhounod D shown in a structural formula (II) in an organic solvent and water;
(2) Adding ruthenium trichloride and sodium periodate, stirring at room temperature for reaction until TLC shows that the reaction is complete, diluting the reaction liquid with an organic solvent, filtering, washing with brine, drying, concentrating in vacuum, and purifying to obtain a colquhounod D derivative 22;
(3) Dissolving a compound colquhounod D shown in a structural formula (II) in an organic solvent;
(4) And adding m-chloroperoxybenzoic acid and sodium bicarbonate under inert gas atmosphere, stirring at room temperature until TLC shows that the reaction is complete, and quenching the reaction solution with sodium bicarbonate solution, extracting, drying, concentrating in vacuum and purifying to obtain the colquhounod D derivatives 37 and 38.
(5) Dissolving the colquhounod D derivative 22 in an organic solvent;
(6) Under the atmosphere of inert gas, adding organic amine (or p-bromophenol or benzyl mercaptan) and condensing agent, stirring at room temperature to react until TLC shows that the reaction is complete, and concentrating and purifying the reaction liquid in vacuum to obtain the colquhounod D derivative 23-30.
(7) Dissolving the colquhounod D derivative 22 in an organic solvent;
(8) DPPA and triethylamine are added under inert gas atmosphere, stirring reaction is carried out at 45 ℃ until TLC shows that the reaction is complete, and the reaction liquid is concentrated and purified in vacuum to obtain the colquhounod D derivative 32.
(9) Dissolving the colquhounod D derivative 23 in an organic solvent;
(10) Adding 1M diisobutyl aluminum hydride in inert gas atmosphere, stirring at 0 ℃ to react until TLC shows complete reaction, quenching the reaction liquid with potassium sodium tartrate, extracting, drying, concentrating in vacuum, and purifying to obtain a colquhounod D derivative 36;
(11) Dissolving the colquhounod D derivative 24 in an organic solvent;
(12) Under the inert gas atmosphere, DPPA is added, heating reflux is carried out, stirring is carried out until TLC shows that the reaction is complete, and the reaction liquid is concentrated and purified in vacuum to obtain the colquhouoid D derivative 31;
(13) Dissolving the colquhounod D derivative 30 in an organic solvent;
(14) Under the inert gas atmosphere, PPh is added 3 AuNTf 2 Stirring at room temperature until TLC shows complete reaction, vacuum concentrating and purifying the reaction liquid to obtain an intermediate, dissolving the intermediate in an organic solvent, adding N-bromosuccinimide or triethylamine tri-hydrofluoric acid or HCl in dioxane or DBU into the intermediate, stirring at room temperature until TLC monitors complete reaction, vacuum concentrating and purifying the reaction liquid to obtain the colquhounod D derivative 33-34.
(15) Dissolving the colquhounod D derivative 30 in an organic solvent and water;
(16) Under the inert gas atmosphere, adding an azide reagent, sodium ascorbate and copper sulfate pentahydrate, stirring at room temperature to react until TLC (thin layer chromatography) shows that the reaction is complete, and diluting the reaction liquid with an organic solvent, extracting, drying, concentrating in vacuum and purifying to obtain the colquhounod D derivative 35.
Preferably, in the above preparation method of the colquhouoid D derivative 22-38, the organic solvent selected from one or more of step (1), step (3), step (5), step (7), step (9), step (11), step (13) and step (15) is selected from N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine and ethanol, further preferably chloroform and acetonitrile, further preferably dichloromethane, further preferably one of dichloromethane, tetrahydrofuran and N, N-dimethylformamide, further preferably acetonitrile, further preferably tetrahydrofuran, further preferably pyridine, further preferably acetonitrile, further preferably ethanol, and further preferably ethanol.
Preferably, in the above preparation method of the colquhouoid D derivative 22-38, the organic amine in the step (6) is selected from one of aniline, 3-aminopyridine, 4-aminopyridine, benzylamine, ethanolamine, morpholine, 2-aminophenylsulfide, propargylamine, glutamic acid, N-methylpiperazine and diethanolamine, and more preferably any one of aniline, ethanolamine, propargylamine, N-methylpiperazine and diethanolamine.
Preferably, in the preparation method of the above-mentioned colquhounod D derivative 22-38, when the amine is aniline, diethanolamine, ethanolamine, N-methylpiperazine, the organic solvent in the step (5) is further preferably dichloromethane; when the amine is propargylamine, the organic solvent of step (5) is further preferably N, N-dimethylformamide; when the reactant is benzyl mercaptan, the organic solvent in step (5) is further preferably N, N-dimethylformamide; when the reactant is p-bromophenol, the organic solvent described in step (5) is further preferably dichloromethane.
Preferably, in the preparation method of the above-mentioned colquhounod D derivative 22-38, the condensing agent in the step (6) is selected from one or more of DMAP, DCC, EDCI, HOBt, HATU and DIPEA, and when the reactant is p-bromophenol or the amine is aniline, DMAP and EDCI are further preferred; when the reactant is benzyl mercaptan or the amine is ethanolamine, DMAP, EDCI, HOBt is further preferred; when the amine is diethanolamine, N-methylpiperazine, further preferred are HATU, DIPEA; when the amine is propargylamine, HATU is further preferred.
Preferably, in the preparation method of the colquhouoid D derivative 22-38, the heating reflux temperature in the step (12) is 100 ℃ to 130 ℃, and more preferably 120 ℃.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the molar volume ratio of the colquhounod D to the organic solvent in the step (1) is 0.37mol/L of chloroform, 0.08mol/L of acetonitrile and 0.04mol/L of water.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the molar ratio of the ruthenium trichloride to the sodium periodate to the colquhounod D is 0.3:30.0:1.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the molar ratio of the m-chloroperoxybenzoic acid to the sodium bicarbonate to the colquhounod D is 2:2:1.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the molar ratio of the p-bromophenol or amine (aniline), EDCI, DMAP and the colquhounod D derivative 22 is 4:4:0.8:1.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the molar ratio of the amine (diethanolamine or N-methylpiperazine), HATU, DIPEA and the colquhounod D derivative 22 is 2:1.2:2:1.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the molar ratio of the benzyl mercaptan or amine (ethanolamine), EDCI, DMAP, HOBt and the colquhounod D derivative 22 is 2.4:3.0:0.8:2.4:1.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the molar ratio of the amine (propargylamine), the HATU and the colquhounod D derivative 22 is 1.2:1.0:1.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the molar ratio of DPPA, triethylamine and the colquhounod D derivative 22 in the step (8) is 1.5:1.5:1.0.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the molar ratio of the diisobutylaluminum hydride to the colquhounod D derivative 23 is 20.0:1.0.
Preferably, in the preparation method of the colquhounod D derivatives 22-38, the molar ratio of DPPA to the colquhounod D derivatives 24 in the step (12) is 2.0:1.
Preferably, in the above preparation method of the colquhounod D derivative 22-38, the PPh 3 AuNTf 2 The molar ratio to the colquhounod D derivative 30 was 0.2:1, the molar ratio of n-bromosuccinimide or triethylamine trihydrofluoride to the intermediate was 1.5:1, and the molar ratio of dbu to the intermediate was 0.4:1.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the volume ratio of the organic solvent to the water in the step (15) is 10:1.
Preferably, in the preparation method of the colquhounod D derivative 22-38, the molar ratio of the azide reagent, sodium ascorbate, copper sulfate pentahydrate and the colquhounod D derivative 30 is 1:0.3:0.1:1.
The invention also discloses a preparation method of the colquhounod D derivative 39-40, which comprises the following steps:
(1) Dissolving a compound colquhounod D shown in a structural formula (II) in an organic solvent;
(2) Under inert gas atmosphere, adding 2M hydrochloric acid, stirring at 45 ℃ until TLC shows that the reaction is complete, quenching the reaction liquid with sodium bicarbonate solution, and then extracting, drying, vacuum concentrating and purifying to obtain the colquhounod D derivative 39-40.
Preferably, in the above preparation method of the colquhouoid D derivative 39-40, the organic solvent in the step (1) is one or more selected from N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine, and ethanol, and more preferably tetrahydrofuran.
The invention also discloses a preparation method of the colquhounod D derivative 41-47, which comprises the following steps:
(1) Dissolving a compound colquhounod D shown in a structural formula (II) in an organic solvent;
(2) Adding NaH and halohydrocarbon under inert gas atmosphere, heating and stirring to react until TLC shows that the reaction is complete, filtering the reaction liquid, collecting filtrate, concentrating in vacuum, and purifying to obtain colquhouoid D derivative 41-44;
(3) Dissolving the colquhounod D derivative 43 in an organic solvent;
(4) Adding 1.5M diisobutyl aluminum hydride in inert gas atmosphere, stirring at 0-60 ℃ to react until TLC shows complete reaction, quenching the reaction liquid with potassium sodium tartrate, extracting, drying, concentrating in vacuum, purifying to obtain an intermediate;
(5) Dissolving the intermediate in an organic solvent;
(6) Adding N, N-thiocarbonyldiimidazole and DMAP under inert gas atmosphere, heating to 60-80 ℃, stirring until TLC shows that the reaction is complete, and vacuum concentrating and purifying the reaction liquid to obtain a colquhounod D derivative 45-46;
(7) Dissolving the colquhounod D derivative 45 in an organic solvent and water;
(8) Under the inert gas atmosphere, adding an azide reagent, sodium ascorbate and copper sulfate pentahydrate, stirring at room temperature to react until TLC (thin layer chromatography) shows that the reaction is complete, and diluting the reaction liquid with an organic solvent, extracting, drying, concentrating in vacuum and purifying to obtain the colquhounod D derivative 47.
Preferably, in the above preparation method of the colquhouoid D derivative 41-47, the organic solvent in the step (1), the step (3), the step (5) and the step (7) is one or more selected from N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine and ethanol, the organic solvent in the step (1) is more preferably tetrahydrofuran, the organic solvent in the step (3) is more preferably tetrahydrofuran, the organic solvent in the step (5) is more preferably toluene, and the organic solvent in the step (7) is more preferably ethanol.
Preferably, in the preparation method of the colquhouoid D derivative 41-47, the heating temperature in the step (2) is 25-60 ℃, more preferably the heating temperature of the derivative 41 is 25 ℃, and the heating temperature of the derivative 42-44 is 60 ℃; the heating temperature in the step (4) is 0-60 ℃, and is more preferably 0 ℃; the heating temperature in the step (6) is 60-80 ℃, and more preferably 80 ℃.
Preferably, in the preparation method of the colquhounod D derivative 41-47, the molar ratio of the sodium hydride, the halogenated hydrocarbon and the colquhounod D in the step (2) is 35.0:30.0:1.0.
Preferably, in the preparation method of the colquhounod D derivative 41-47, the molar ratio of 1.5M DIBAl-H to the colquhounod D derivative 43 is 20:1.
Preferably, in the preparation method of the colquhounod D derivative 41-47, the molar ratio of the N, N-thiocarbonyldiimidazole, the DMAP and the colquhounod D derivative 43 is 2:3:1.
Preferably, in the preparation method of the colquhounod D derivative 41-47, the volume ratio of the organic solvent to the water in the step (5) is 10:1.
Preferably, in the preparation method of the colquhounod D derivative 41-47, the molar ratio of the azide reagent, sodium ascorbate, copper sulfate pentahydrate and the colquhounod D derivative 45 is 1:0.3:0.1:1.
The precursor compound with immunosuppressive activity can be obtained through the technical scheme.
The invention also provides application of the colquhouund D derivative in preparing anti-inflammatory drugs or immunosuppression drugs and in preparing drugs for treating ulcerative colitis.
The invention also provides a pharmaceutical composition which comprises any one or any combination of pharmaceutically acceptable pharmaceutical excipients and the colquhouoid D derivative of the dioxane sesterterpene.
The pharmaceutical composition comprises granules, powder fog, spray, patch, tablet, capsule, medicinal granules, oral liquid preparation and injection.
In addition, the application of the pharmaceutical composition in preparing anti-inflammatory drugs or immunosuppression drugs is also provided. And application in preparing medicines for treating ulcerative colitis.
The colquhounod D derivatives 1-47 (compounds 1-47) of the present invention can be used as a medicament directly or in the form of a pharmaceutical composition. The pharmaceutical composition contains 0.1-99%, preferably 0.5-90% of the compound of the invention, the balance being pharmaceutically acceptable, non-toxic and inert pharmaceutically acceptable carriers and/or excipients for humans and animals.
The pharmaceutically acceptable carriers or excipients are one or more solid, semi-solid and liquid diluents, fillers and pharmaceutical formulation adjuvants. The pharmaceutical composition of the present invention is used in the form of a unit weight dose. The medicine of the present invention may be administered in various forms, including liquid preparation, solid preparation, injection, external preparation, spray and compound preparation.
Compared with the prior art, the invention has the following advantages:
1. the invention provides a new colquhouund D derivative 1-47 (compound 1-47), which fills the blank of the prior art.
2. The invention provides a method for preparing the colquhounod D derivative 1-47, which has the advantages of easily available raw materials, easy operation and high yield, and is suitable for industrial production.
3. The invention provides a pharmaceutical composition taking the colquhouund D derivative 1-47 as an active ingredient, and provides a new medicine with better medicinal effect for new anti-inflammatory medicines and immunosuppression medicines.
4. In vitro immunosuppressive activity screening of mouse T cells is carried out on the dioxane sesterterpene colquhounod D derivatives 1-47, and 28 compounds with enhanced IFN-gamma secretion inhibiting activity in the mouse T cells are found compared with the prototype compound colquhounod D. And the compound does not show obvious toxicity, which indicates that the compound has stronger immunosuppressive activity research value. In vitro immunosuppressive activity screening of the BMDM cells of the mice was performed on the compounds 1-47, and found that 37 compounds with enhanced secretion inhibition of inflammatory factor IL-1 beta secreted by the BMDM cells stimulated by the LPS+ATP of the mice compared with the prototype natural product colquhouoid D, and did not show obvious toxicity. The compounds have outstanding immunosuppressive activity.
5. The colquhounod D derivatives 1-47 can be used as medicines for treating related diseases. Can be used for preparing anti-inflammatory drugs, immunosuppression drugs and ulcerative colitis treatment drugs.
Drawings
FIG. 1 is a schematic diagram of the structural general formula of the colquhounod D derivative 1-47 (compound 1-47) of the present invention. In the general formula (I), R 1 Substituents include, but are not limited to, hydrogen, methyl, n-propyl, n-butyl, n-pentyl, allyl, propargyl, benzoyl, acetyl, R 2 Substituents include, but are not limited to, carboxyl, hydroxymethyl,/-> R 3 And R is R 4 Substituents include, but are not limited to, substituted or unsubstituted cyclopropane, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, hydrogen atom; r is R 5 Substituents include, but are not limited to, hydrogen, methyl, allyl, propargyl, benzyl, biotin; r is R 6 Substituents include, but are not limited to, hydroxy.
FIG. 2 is a graphic illustration of the colquhounod D derivative 8 (Compound 8) of the present invention as a colquhounod for alleviating DSS-induced colitis in mice.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
Preparation of colquhounod D: pulverizing dried aerial parts of the torch (11 kg), and extracting with methanol for 24 hr for 3 times. Evaporating the organic solvent to obtain methanol extract, diluting with water, extracting with ethyl acetate for 3 times, and evaporating the organic solvent to obtain ethyl acetate extract 1.2kg. The ethyl acetate extract was then eluted with chloroform/acetone (1:0, 9:1, 8:2, 7:3, 1:1, 0:1) by silica gel column chromatography to give 6 fractions (fractions A-F). Fr.A (437.4 g) was eluted with methanol/water (80% -100%) by macroporous resin column chromatography to give Fr.A1-A3.Fr.A2 (149.4 g) was eluted with petroleum ether/chloroform (1:1) by silica gel column chromatography to give Fr.A2-1-A2-3.Fr.A2-1 (100 g) was eluted with methanol water (60% -100%) by macroporous resin column chromatography to give Fr.A2-1-1-A2-1-5.Fr.A2-1-4-A2-1-5 (60.1 g) was subjected to silica gel column chromatography and RP-C18 column chromatography to give colquhounod D (250 mg).
Preparation of derivatives 1-3:
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under the protection of argon, colquhounod D (1.0 eq) was dissolved with dry THF, cooled to 0 ℃, and then DIBAl-H in PhMe (20.0 eq) was slowly added dropwise to the reaction solution. After the completion of the dropwise addition, the reaction temperature was raised to 60℃and the reaction was stirred for 3 hours. After the reaction is finished, the reaction solution is dripped into a saturated potassium sodium tartrate solution to quench, the aqueous phase is extracted by ethyl acetate, and the organic phases are combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (PE/EA 8:1-1:1) to give derivatives 1, 2 and 3 in 48%, 13% and 32%, respectively.
Derivative 1: c (C) 25 H 36 O 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.25(q,J＝3.4,2.4Hz,1H),6.15(d,J＝2.1Hz,1H),4.95(m,1H),4.06(t,J＝2.8Hz,1H),3.74(dd,J＝5.7,3.4Hz,1H),3.06(d,J＝2.3Hz,1H),2.81(s,1H),2.62(ddd,J＝9.8,6.6,2.9Hz,2H),2.52(dd,J＝9.7,2.5Hz,1H),2.28(dq,J＝6.7,3.9Hz,1H),2.23(dt,J＝10.2,2.7Hz,1H),2.05(m,1H),1.92(d,J＝2.5Hz,3H),1.85(ddd,J＝12.8,7.1,3.8Hz,3H),1.63(d,J＝2.4Hz,3H),1.21–1.10(m,4H),1.05(dd,J＝6.8,2.6Hz,3H),1.00(dd,J＝5.8,2.6Hz,3H),0.97–0.88(m,2H),0.83(ddd,J＝8.3,5.1,3.3Hz,1H),0.20(ddt,J＝8.2,5.9,2.8Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ152.3,140.4,113.7,113.5,108.8,86.8,83.9,79.4,69.3,58.9,48.1,47.0,46.3,43.0,33.9,33.6,26.8,24.1,22.1,19.6,19.1,16.0,11.9,9.8,8.8；HRMS(ESI)at m/z 439.2458[M+Na] + (calcd.for C 25 H 36 O 5 Na,m/z 439.2455).
Derivative 2: c (C) 25 H 38 O 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝1.8Hz,1H),6.16(d,J＝1.9Hz,1H),4.14(dt,J＝6.6,3.5Hz,1H),3.92(dt,J＝5.1,2.5Hz,1H),3.82(d,J＝6.1Hz,1H),3.71(d,J＝5.3Hz,1H),3.42(s,1H),3.20(d,J＝8.8Hz,1H),2.68–2.55(m,2H),2.52(dq,J＝11.2,6.4Hz,1H),2.27(ddt,J＝12.7,8.3,6.3Hz,1H),2.14(ddd,J＝13.5,11.1,5.2Hz,1H),2.01(td,J＝11.6,4.1Hz,1H),1.93(s,3H),1.87(dd,J＝12.4,6.3Hz,1H),1.76(ddd,J＝13.6,11.2,6.0Hz,1H),1.73(d,J＝2.5Hz,1H),1.60(ddd,J＝11.3,8.3,2.6Hz,1H),1.44(s,3H),1.21(m,1H),1.09(d,J＝6.5Hz,3H),1.06(d,J＝6.6Hz,3H),1.03(d,J＝6.0Hz,3H),0.87(tt,J＝9.0,4.6Hz,1H),0.75(dddd,J＝11.8,10.2,7.1,4.8Hz,1H),0.54(dt,J＝8.3,4.6Hz,1H),0.23(dt,J＝8.1,4.7Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ152.5,140.5,113.9,113.7,93.9,86.0,82.8,73.2,66.8,66.0,52.1,48.8,44.9,40.9,33.2,31.7,22.4,22.1,21.3,20.1,18.9,16.1,13.6,12.1,10.0；HRMS(ESI)at m/z 441.2613[M+Na] + (calcd.for C 25 H 38 O 5 Na,m/z 441.2611).
Derivative 3: c (C) 25 H 38 O 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.27(d,J＝1.8Hz,1H),6.17(d,J＝1.9Hz,1H),4.13(dt,J＝6.4,3.1Hz,1H),4.08(d,J＝6.1Hz,1H),4.06(d,J＝6.0Hz,1H),3.88(dt,J＝5.9,2.9Hz,1H),3.11(d,J＝7.7Hz,1H),3.09(s,1H),2.75–2.63(m,2H),2.25(dq,J＝13.7,6.7Hz,1H),2.06(m,2H),1.94(s,3H),1.87(ddt,J＝13.9,10.3,7.0Hz,3H),1.79(d,J＝3.3Hz,1H),1.71(ddd,J＝10.7,7.8,2.4Hz,1H),1.31(s,3H),1.24(td,J＝12.4,3.2Hz,1H),1.09(d,J＝6.1Hz,3H),1.06(d,J＝6.7Hz,3H),1.03(d,J＝6.0Hz,3H),0.91(tt,J＝8.4,4.7Hz,1H),0.73(m,1H),0.59(dt,J＝8.6,4.5Hz,1H),0.27(dt,J＝8.9,4.6Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ152.3,140.6,114.0,113.7,89.2,83.7,81.6,72.8,67.2,66.4,52.2,49.4,45.1,43.4,34.3,33.1,22.3,21.6,20.4,19.7,18.9,14.5,12.8,12.0,10.0；HRMS(ESI)at m/z 441.2613[M+Na] + (calcd.for C 25 H 38 O 5 Na,m/z441.2611).
Example 2
Preparation of derivative 4:
the preparation of derivative 4 was carried out according to the preparation method of derivatives 1-3, with a reaction temperature of 0℃and a yield of 87%.
Derivative 4: c (C) 25 H 36 O 5 ,white powder. 1 H NMR(500MHz,Acetone-d 6 )δ7.29(d,J＝1.9Hz 1H),6.18(d,J＝1.9Hz 1H),4.83(s,1H),4.29(s,1H),4.11(t,J＝6.8Hz1H),3.86(dd,J＝2.4,1.0Hz,1H),2.72(ddd,J＝14.2,10.7,5.6Hz,2H),2.20(s,1H),2.15(tt,J＝6.6,1.3Hz,1H),2.03-1.92(m,3H),1.95(s,3H),1.92(m,1H),1.86(m,1H),1.76(dd,J＝6.6,2.4Hz,1H),1.42(s,3H),1.38(m,1H),1.26(dt,J＝8.4,4.9Hz,1H),1.17(d,J＝6.9Hz,3H),0.97(d,J＝6.0Hz,3H),0.87(m,4H),0.75(ddd,J＝8.4,5.3,3.4Hz,1H),0.02(ddd,J＝8.5,5.2,3.4Hz,1H)； 13 C NMR(126MHz,Acetone-d 6 )δ152.2,140.7,114.1,113.8,110.4,87.3,80.7,80.0,74.4,52.9,52.3,44.6,43.4,42.8,37.7,37.3,26.6,24.4,23.3,20.9,19.1,15.7,10.7,10.3,9.9；HRMS(ESI)at m/z 439.2458[M+Na] + (calcd.for C 25 H 36 O 5 Na,m/z 439.2460).
Example 3
Preparation of derivative 5:
derivative 1 (1.0 eq) was dissolved in dry DCM under the protection of argon, then DMAP (1.0 eq), TEA (31.6 eq) and acetic anhydride (41.2 eq) were added sequentially to the reaction system and the reaction was heated to 45 ℃ and stirred for 4h. After the complete consumption of the starting material was monitored by TLC, a saturated sodium bicarbonate solution was slowly added dropwise to the reaction solution to quench the reaction, the aqueous phase was extracted with diethyl ether and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (PE/EA 30:1-10:1) to give derivative 5 in 98% yield.
Derivative 5: c (C) 27 H 38 O 6 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.25(d,J＝1.8Hz,1H),6.15(d,J＝1.8Hz,1H),5.81(t,J＝9.6Hz,1H),5.62(s,1H,CH 2 Cl 2 ),4.10(d,J＝2.7Hz,1H),3.32(s,1H),2.77(m,1H),2.67(d,J＝9.9Hz,1H),2.58(ddd,J＝14.5,11.8,5.4Hz,1H),2.49(ddd,J＝14.5,11.7,5.3Hz,1H),2.36(qd,J＝7.1,2.7Hz,1H),2.31(dt,J＝10.6,2.7Hz,1H),2.19(td,J＝9.9,5.3Hz,1H),2.08(s,3H),1.91(s,3H),1.88(m,1H),1.86–1.78(m,2H),1.56(s,3H),1.22(d,J＝7.1Hz,3H),1.19(m,1H),1.01(t,J＝6.6Hz,6H),0.96(dt,J＝8.8,4.8Hz,1H),0.92(dd,J＝9.8,4.2Hz,1H),0.86(m,1H),0.23(ddd,J＝8.8,5.3,3.7Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ170.5,152.0,140.6,114.0,113.7,109.0,86.7,84.2,79.3,71.9,56.5,48.0,45.9,43.8,43.0,35.0,33.5,26.8,23.3,22.0,21.2,19.6,19.2,16.1,12.2,9.9,8.9；HRMS(ESI)at m/z 481.2565[M+Na] + (calcd.for C 27 H 38 O 6 Na,m/z481.2561).
Example 4
Preparation of derivatives 6-7
Preparation of compound 6 reference compound 5 was synthesized in 57% yield.
Under the protection of argon, P is 2 S 5 (1.0 eq) was added to a solution of sodium bicarbonate (1.0 eq) in THF. After the mixture was stirred at 25 ℃ to clarify, it was cooled to 0 ℃, and derivative 6 (1.0 eq) was added to the clarified solution, stirring was continued at 0 ℃ for 30min, and then the reaction solution was slowly raised to 80 ℃ to stir for 48h. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/EA 10:1-1:1) to give compound 7 in 71% yield.
Derivative 6: c (C) 32 H 40 O 6 ,colorless oil. 1 H NMR(400MHz,Acetone-d 6 )δ8.08(d,J＝7.1Hz,2H),7.66(t,J＝7.4Hz,1H),7.56(t,J＝7.6Hz,2H),7.19(d,J＝1.9Hz,1H),6.13(d,J＝9.4Hz,1H),6.10(s,1H),4.16(s,1H),3.40(s,1H),3.01(m,1H),2.88(d,J＝10.0Hz,1H),2.48–2.43(m,2H),2.41(m,1H),2.38(d,J＝2.4Hz,1H),,2.33(td,J＝10.1,9.4,4.9Hz,1H),1.95(m,1H),1.89–1.76(m,5H),1.60(s,3H),1.30(d,J＝7.2Hz,3H),1.24(dt,J＝14.5,3.1Hz,1H),1.06(d,J＝6.8Hz,3H),1.04(d,J＝5.8Hz,3H),1.00(dd,J＝8.6,4.8Hz,1H),0.95(m,1H),0.88(m,1H),0.25(m,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ166.4,151.8,140.6,134.2,131.4,130.3,129.7,113.9,113.6,109.1,86.6,84.4,79.3,72.9,56.6,48.1,45.7,44.2,43.0,35.4,33.6,27.0,23.4,22.0,19.6,19.2,16.1,12.2,9.8,9.0；HRMS(ESI)at m/z 543.2716
[M+Na] + (calcd.for C 32 H 40 O 6 Na,m/z 543.2717).
Derivative 7: c (C) 32 H 42 O 6 S,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ8.10(d,J＝7.4Hz,2H),7.62(t,J＝7.4Hz,1H),7.49(t,J＝7.7Hz,2H),7.27(d,J＝1.8Hz,1H),6.16(d,J＝1.7Hz,1H),5.72(d,J＝3.2Hz,1H),4.29(s,1H),3.49(d,J＝2.9Hz,1H),3.16(s,1H),2.88(m,1H),2.84(m,1H),2.81(s,1H),2.60(m,1H),2.51(m,1H),2.37(t,J＝10.3Hz,1H),2.28(m,1H),2.21(s,1H),2.11(m,2H),1.97(s,3H),1.99-1.92(m,2H),1.83-1.75(m,2H),1.71(m,1H),1.27(d,J＝7.3Hz,1H),1.22(d,J＝6.4Hz,3H),1.16(s,3H),1.04(d,J＝6.4Hz,3H),0.98(d,J＝6.5Hz,3H)； 13 C NMR(151MHz,Acetone-d 6 )δ167.5,152.3,140.6,133.8,132.1,130.8,129.2,114.6,114.2,113.7,86.7,83.7,72.1,69.6,61.3,52.4,51.3,47.0,45.3,41.8,39.7,38.0,33.6,33.5,22.4,22.1,18.9,18.6,13.8,10.1；HRMS(ESI)at m/z 577.2595[M+Na] + (calcd.for C 32 H 42 O 6 SNa,m/z 577.2600).
Example 5
Preparation of derivatives 8-9:
derivative 1 (1.0 eq), N-thiocarbonylimidazole (2.0 eq) and DMAP (3.0 eq) were dissolved in dry toluene under argon and then heated to 80℃and reacted with stirring. After 3 hours of reaction, the reaction mixture was cooled to room temperature, and N, N-thiocarbonylimidazole (4.3 eq) and DMAP (6.3 eq) were added thereto and the reaction was continued until 80 ℃. After the reaction was carried out for 5 hours, the reaction mixture was cooled to room temperature again, and N, N-thiocarbonylimidazole (4.3 eq) and DMAP (6.3 eq) were added thereto to continue the reaction at 80 ℃. The reaction was carried out for 15 hours and then the same procedure was followed. After the reaction had proceeded for 19 hours, N-thiocarbonylimidazole (4.3 eq) and DMAP (6.3 eq) were added again and the reaction continued at 80℃until the reaction was complete as indicated by thin layer chromatography. After the reaction, the mixture was concentrated in vacuo and purified by silica gel column chromatography (PE/EA 6:1) to give derivatives 8 and 9 in 47% and 10% yields, respectively.
Derivative 8: c (C) 29 H 38 O 5 S,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ8.38(s,1H),7.76(t,J＝1.5Hz,1H),7.19(d,J＝1.8Hz,1H),7.08(d,J＝1.8Hz,1H),6.62(m,1H),6.12(d,J＝1.8Hz,1H),4.20(s,1H),3.57(s,1H),3.05(d,J＝10.1Hz,1H),2.99(dd,J＝11.4,4.8Hz,1H),2.50(dd,J＝9.8,7.3Hz,2H),2.44(m,3H),1.98(m,1H),1.94(m,1H),1.85(s,3H),1.82(m,1H),1.51(s,3H),1.34–1.26(m,4H),1.08(d,J＝6.8Hz,3H),1.04(d,J＝5.9Hz,3H),1.01(dd,J＝8.8,4.6Hz,1H),0.95(m,1H),0.89(m,1H),0.27(ddd,J＝8.8,5.3,3.7Hz,1H)； 13 C NMR(151MHz,Acetone)δ185.1,151.7,140.6,137.9,132.2,119.0,114.0,113.6,109.2,86.4,84.7,83.0,79.3,55.8,48.1,45.4,43.8,43.0,35.9,33.4,26.9,23.2,22.0,19.5,19.2,16.2,12.3,9.8,9.0；HRMS(ESI)at m/z 549.2398[M+Na] + (calcd.for C 29 H 38 O 5 SNa,m/z 549.2394).
Derivative 9: c (C) 29 H 38 N 2 O 6 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ8.17(s,1H),7.55(s,1H),7.20(d,J＝1.9Hz,1H),7.08(s,1H),6.12(d,J＝1.8Hz,1H),6.08(t,J＝9.2Hz,1H),4.17(s,1H),3.51(s,1H),2.95(dd,J＝11.4,6.2Hz,1H),2.91(d,J＝10.1Hz,1H),2.52(td,J＝8.2,7.4,3.3Hz,2H),2.47–2.39(m,3H),1.96(t,J＝6.6Hz,1H),1.95–1.82(m,2H),1.86(s,3H),1.56(s,3H),1.27(d,J＝6.9Hz,4H),1.07(d,J＝6.8Hz,3H),1.03(d,J＝5.9Hz,3H),1.00(dt,J＝9.0,4.7Hz,1H),0.95(m,1H),0.87(m,1H),0.26(dt,J＝8.9,4.5Hz,1H)； 13 C NMR(151MHz,Acetone)δ151.78,149.3,140.6,137.9,131.7,118.1,114.0,113.6,109.2,86.5,84.5,79.3,77.6,56.6,48.1,45.6,43.9,43.0,35.4,33.4,26.8,23.3,21.9,19.5,19.2,16.1,12.3,9.8,9.0；HRMS(ESI)at m/z 533.2620[M+Na] + (calcd.for C 29 H 38 N 2 O 6 Na,m/z 533.2622 Example 6):
preparation of derivatives 10 and 11:
under the protection of argon, colquhounod D (1.0 eq) was dissolved in dry THF, then sodium hydride (30.0 eq) and methyl isothiocyanate (35.0 eq) were added in sequence, and the reaction solution was stirred at room temperature for 12h. After TLC monitoring the complete consumption of the starting material, the reaction solution was quenched by dropwise addition of an aqueous ammonium chloride solution, the aqueous phase was extracted with ethyl acetate, the organic phase was concentrated under reduced pressure to remove the organic solvent, the residue was purified by column chromatography on silica gel (PE/EA 25:1-5:1) to give the crude product, which was purified by semi-preparative HPLC to give the derivatives 10 and 11 in 64% yield.
Derivative 10: c (C) 27 H 39 NO 5 S,white powder. 1 H NMR(600MHz,Acetone-d 6 )δ8.25(s,1H),7.24(d,J＝1.9Hz,1H),6.50(t,J＝9.6Hz,1H),6.14(d,J＝1.9Hz,1H),4.11(s,1H),3.34(s,1H),3.03(d,J＝4.7Hz,3H),2.81(d,J＝9.9Hz,1H),2.79(d,J＝8.9Hz,1H),2.58–2.45(m,2H),2.35(dtd,J＝14.2,7.1,2.5Hz,1H),2.31(dt,J＝10.4,2.5Hz,1H),2.23(qd,J＝9.6,4.8Hz,1H),1.91(d,J＝3.6Hz,3H),1.90–1.79(m,3H),1.51(s,3H),1.25(d,J＝7.1Hz,3H),1.19(ddd,J＝12.9,10.7,2.7Hz,1H),1.01(d,J＝5.8Hz,3H),0.98(d,J＝6.8Hz,3H),0.96(m,1H),0.91(m,1H),0.84(m,1H),0.22(ddd,J＝8.7,5.1,3.6Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ191.4,152.1,140.6,113.9,113.6,109.0,86.8,84.4,79.3,77.3,56.0,48.0,45.8,43.9,43.0,35.2,33.5,32.3,26.8,23.4,22.0,19.6,19.2,16.1,12.1,9.93,8.93；HRMS(ESI)at m/z 512.2441[M+Na] + (calcd.for C 27 H 39 NO 5 SNa,m/z 512.2441).
Derivative 11: c (C) 27 H 39 NO 5 S,white powder. 1 H NMR(600MHz,Acetone-d 6 )δ8.20(s,1H),7.24(d,J＝1.9Hz,1H),6.50(t,J＝9.6Hz,1H),6.14(d,J＝1.9Hz,1H),4.14(s,1H),3.37(s,1H),3.03(d,J＝4.7Hz,3H),2.92(d,J＝10.0Hz,1H),2.89(d,J＝8.9Hz,1H),2.65–2.56(m,2H),2.35(dtd,J＝14.2,7.1,2.5Hz,1H),2.31(dt,J＝10.4,2.5Hz,1H),2.23(qd,J＝9.6,4.8Hz,1H),1.91(d,J＝3.6Hz,4H),1.90–
1.79(m,2H),1.57(s,3H),1.25(d,J＝7.1Hz,3H),1.19(m,1H),1.07(d,J＝6.8Hz,3H),1.01(d,J＝5.8Hz,3H),0.96(m,1H),0.91(m,1H),0.84(m,1H),0.22(ddd,J＝8.7,5.1,3.6Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ190.2,152.1,140.6,113.9,113.6,109.0,86.5,84.5,79.3,79.1,56.1,48.0,45.8,44.0,43.0,35.6,33.5,32.1,26.8,23.4,22.1,19.6,19.2,16.1,12.2,9.93,8.93；HRMS(ESI)at m/z 512.2441[M+Na] + (calcd.for C 27 H 39 NO 5 SNa,m/z 512.2441).
Example 7
Preparation of derivatives 12 and 13:
preparation of derivatives 12 and 13 reference the synthetic method of derivatives 10 and 11 in 23% yield.
Derivative 12: c (C) 32 H 41 NO 5 S,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ10.27(d,J＝40.2Hz,1H),7.71(d,J＝8.1Hz,1H),7.36(t,J＝7.8Hz,2H),7.25(d,J＝6.1Hz,1H),7.20(t,J＝7.5Hz,2H),6.60(dt,J＝19.5,9.5Hz,1H),6.14(d,J＝8.6Hz,1H),4.13(d,J＝12.9Hz,1H),3.56(m,1H),2.90–2.85(m,2H),2.62(m,1H),2.36(dt,J＝12.3,5.1Hz,3H),2.26(dd,J＝9.8,5.2Hz,1H),1.89(d,J＝13.2Hz,3H),1.86(d,J＝8.6Hz,3H),1.57(s,3H),1.25(t,J＝7.6Hz,3H),1.20(m,1H),1.10(d,J＝6.8Hz,3H),1.05(d,J＝6.7Hz,3H),1.00(dd,J＝17.2,6.1Hz,1H),0.88–0.77(m,2H),0.20(m,1H). 13 C NMR(151MHz,Acetone-d 6 )δ188.8,151.8,140.6,139.7,129.3,126.4,124.6,113.9,113.5,109.0,86.7,84.3,79.2,76.9,55.6,47.9,45.8,43.8,42.8,35.3,33.4,26.9,23.6,22.0,19.4,19.1,16.2,12.1,9.9,8.9；HRMS(ESI)at m/z574.2600[M+Na] + (calcd.for C 32 H 41 NO 5 SNa,m/z 574.2603).
Derivative 13: c (C) 32 H 41 NO 5 S,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ10.27(d,J＝40.2Hz,1H),7.40(d,J＝4.8Hz,3H),7.25(d,J＝6.1Hz,3H),6.60(dt,J＝19.5,9.5Hz,1H),6.14(d,J＝8.6Hz,1H),4.13(d,J＝12.9Hz,1H),3.56(m,1H),2.87(m,1H),2.73(s,1H),2.56(m,1H),2.36(dt,J＝12.3,5.1Hz,4H),1.89(d,J＝13.2Hz,3H),1.86(d,J＝8.6Hz,2H),1.74(m,1H),1.25(t,J＝7.6Hz,3H),1.20(m,1H),1.12(d,J＝3.4Hz,3H),1.10(d,J＝6.8Hz,3H),1.05(d,J＝6.7Hz,3H),1.00(dd,J＝17.2,6.1Hz,1H),0.83(m,2H),0.21(m,1H). 13 C NMR(151MHz,Acetone-d 6 )δ188.8,151.8,140.6,139.7,129.8,126.9,124.7,113.9,113.5,109.0,86.1,84.3,80.6,79.2,55.6,48.0,45.7,43.9,43.0,35.6,33.2,26.5,23.8,22.0,19.3,19.1,16.0,12.1,9.9,8.9；HRMS(ESI)at m/z 574.2600[M+Na] + (calcd.for C 32 H 41 NO 5 SNa,m/z 574.2603).
Example 8
Preparation of derivatives 14 and 15:
preparation of derivatives 14 and 15 reference the synthesis of derivatives 10 and 11 at a reaction temperature of 45℃with a yield of 36%.
Derivative 14: c (C) 31 H 40 N 2 O 5 S,yellow oil. 1 H NMR(600MHz,Acetone-d 6 )δ10.41(s,1H),,8.78–8.74(m,1H),8.40(d,J＝4.6Hz,1H),8.22(d,J＝8.9Hz,1H),7.40(dd,J＝8.3,4.8Hz,1H),7.25(s,1H),6.57(t,J＝9.6Hz,1H),6.14(s,1H),4.13(d,J＝10.1Hz,1H),3.57(s,1H),2.92(m,1H),2.88(d,J＝10.1Hz,1H),,2.63–2.50(m,2H),2.40–2.34(m,2H),2.29(m,1H),1.89(d,J＝13.7Hz,6H),1.57(s,3H),1.29–1.18(m,4H),1.05(d,J＝6.7Hz,3H),1.03–0.96(m,4H),0.92(s,1H),0.85(m,1H),0.23(dd,J＝11.2,6.2Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ189.6,151.8,147.4,146.2,140.6,136.4,131.7,124.0,113.9,113.5,109.0,86.6,84.3,79.2,77.7,55.6,47.9,45.7,43.7,42.8,35.3,33.4,26.8,23.5,22.0,19.4,19.1,16.1,12.1,9.9,8.9；HRMS(ESI)at m/z 575.2550[M+Na] + (calcd.for C 31 H 40 N 2 O 5 S Na,m/z575.2550).
Derivative 15: c (C) 31 H 40 N 2 O 5 S,yellow oil. 1 H NMR(600MHz,Acetone-d 6 )δ10.35(s,1H),8.65(s,1H),8.45(s,1H),7.83(d,J＝8.5Hz,1H),7.44(s,1H),7.25(s,1H),6.60(s,1H),6.14(s,1H),4.13(d,J＝10.1Hz,1H),3.57(s,1H),2.92(m,1H),2.70(s,1H),2.47(d,J＝14.4Hz,1H),2.40–2.34(m,3H),2.29(m,1H),1.89(d,J＝13.7Hz,5H),1.78(s,1H),1.29–1.18(m,4H),1.14(s,3H),1.05(d,J＝6.7Hz,3H),1.03–0.96(m,4H),0.92(s,1H),0.84(m,1H),0.23(dd,J＝11.2,6.2Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ189.6,151.8,147.8,145.9,140.6,136.4,131.6,124.5,113.9,113.5,109.0,86.0,84.2,81.2,79.2,55.5,48.0,45.6,43.9,43.0,35.7,33.2,26.5,23.7,22.0,19.3,19.1,16.0,12.1,9.9,8.9；HRMS(ESI)at m/z 575.2550[M+Na] + (calcd.for C 31 H 40 N 2 O 5 S Na,m/z 575.2550).
Example 9
Preparation of derivative 16:
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preparation of derivative 16 referring to the synthetic methods of derivatives 10 and 11, the reaction temperature was 45 ℃, without purification by semi-preparative HPLC, with a 73% yield.
Derivative 16: c (C) 27 H 40 O 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝1.9Hz,1H),6.16(d,J＝1.9Hz,1H),4.33(t,J＝9.2Hz,1H),4.04(s,1H),3.45(s,3H),3.39(s,3H),2.76(dtd,J＝14.5,7.1,5.1Hz,1H),2.69(qd,J＝6.9,3.1Hz,1H),2.63(ddd,J＝14.7,11.9,5.0Hz,1H),2.60–2.52(m,2H),2.34(dt,J＝10.4,2.8Hz,1H),2.25(ddd,J＝10.4,9.0,5.1Hz,1H),1.94(s,3H),1.91(m,1H),1.83(m,1H),1.80(m,1H),1.49(s,3H),1.19(d,J＝6.9Hz,3H),1.16(m,1H),1.06(d,J＝6.8Hz,3H),1.04–1.01(m,5H),0.92(m,1H),0.28(m,1H). 13 C NMR(151MHz,Acetone-d 6 )δ152.5,140.5,113.8,113.6,108.2,89.2,85.8,80.2,78.6,58.6,57.0,52.1,48.6,45.5,44.6,43.2,34.1,28.3,26.5,23.8,22.0,21.6,19.2,16.6,13.2,11.3,9.9；HRMS(ESI)at m/z 467.2768[M+Na] + (calcd.for C 27 H 40 O 5 Na,m/z 467.2768).
Example 10
Preparation of derivative 17:
preparation of derivative 17 referring to the synthetic methods of derivatives 10 and 11, the reaction temperature was 45 ℃, without purification by semi-preparative HPLC, with a yield of 36%.
Derivative 17: c (C) 28 H 42 O 5 ,yellow oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(s,1H),6.16(s,1H),4.45(t,J＝9.1Hz,1H),4.06(s,1H),3.50(tq,J＝6.4,2.6Hz,2H),3.14(s,1H),2.79(m,1H),2.68–2.56(m,3H),2.29(m,1H),2.26(q,J＝4.9Hz,1H),2.22(td,J＝9.9,9.4,4.8Hz,1H),1.93(s,3H),1.92(m,1H),1.85(dt,J＝12.7,6.2Hz,2H),1.63–1.56(m,2H),1.57(s,3H),1.19(d,J＝7.0Hz,3H),1.15(ddd,J＝12.9,10.4,2.6Hz,1H),1.06(d,J＝6.6Hz,3H),1.01(d,J＝5.9Hz,3H),0.96(t,J＝6.8Hz,3H),0.93(m,1H),0.90(m,1H),0.84(m,1H),0.21(m,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ152.3,140.5,113.8,113.6,108.9,86.6,84.0,79.4,78.4,72.8,57.3,48.0,46.1,44.4,43.2,34.3,33.6,26.7,24.3,23.8,22.2,19.6,19.2,16.1,12.1,11.4,9.9,8.9；HRMS(ESI)at m/z 481.2927[M+Na] + (calcd.for C 28 H 42 O 5 Na,m/z481.2930).
Example 11
Preparation of derivative 18
Preparation of derivative 18 referring to the synthetic methods of derivatives 10 and 11, the reaction temperature was 45 ℃, without purification by semi-preparative HPLC, with a yield of 72%.
Derivative 18: c (C) 29 H 44 O 5 ,yellow oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝1.9Hz,1H),6.16(d,J＝1.9Hz,1H),4.45(t,J＝9.1Hz,1H),4.06(s,1H),3.54(q,J＝6.2Hz,2H),3.14(s,1H),2.78(m,1H),2.69–2.55(m,3H),2.29(m,1H),2.25(d,J＝2.7Hz,1H),2.21(td,J＝10.1,9.4,4.9Hz,1H),1.93(s,3H),1.91(m,1H),1.84(dt,J＝13.0,6.6Hz,2H),1.56(s,3H),1.60–1.52(m,2H),1.48–1.39(m,2H),1.19(d,J＝7.0Hz,3H),1.16(m,1H),1.06(d,J＝7.0Hz,3H),1.00(d,J＝5.9Hz,3H),0.92(qd,J＝8.4,7.4,2.7Hz,5H),0.84(dt,J＝8.5,4.3Hz,1H),0.21(m,1H)； 13 CNMR(151MHz,Acetone-d 6 )δ152.3,140.5,113.8,113.6,108.9,86.6,84.0,79.4,78.5,70.9,57.3,48.0,46.1,44.4,43.2,34.3,33.6,33.3,26.7,23.8,22.2,20.3,19.6,19.2,16.1,14.3,12.1,9.9,8.9；HRMS(ESI)at m/z 495.3087[M+Na] + (calcd.for C 29 H 44 O 5 Na,m/z 495.3086).
Example 12
Preparation of derivative 19
Preparation of derivative 19 referring to the synthetic methods of derivatives 10 and 11, the reaction temperature was 45 ℃, without purification by semi-preparative HPLC, with a yield of 27%.
Derivative 19: c (C) 30 H 46 O 5 ,yellow oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝1.9Hz,1H),6.16(d,J＝1.9Hz,1H),4.45(t,J＝9.1Hz,1H),4.06(s,1H),3.59–3.49(m,2H),3.14(s,1H),2.79(m,1H),2.69–2.56(m,3H),2.29(m,1H),2.25(s,1H),2.22(m,1H),1.93(s,3H),1.92–1.81(m,3H),1.61–1.57(m,2H),1.56(s,3H),1.43–1.36(m,2H),1.36–1.27(m,2H),1.19(d,J＝6.9Hz,3H),1.15(m,1H),1.06(d,J＝6.7Hz,3H),1.01(d,J＝5.8Hz,3H),0.94(dq,J＝9.6,4.7Hz,1H),0.89(q,J＝7.4,7.0Hz,4H),0.84(m,1H),0.21(m,1H)； 13 C NMR(151MHz,Acetone)δ152.3,140.5,114.5,113.8,113.6,108.9,86.6,84.0,79.4,78.5,71.1,57.3,48.0,46.1,44.4,43.2,34.3,33.6,29.4,26.7,23.8,23.3,22.2,19.6,19.2,16.1,14.4,12.1,9.9,8.9；HRMS(ESI)at m/z 509.3239[M+Na] + (calcd.for C 30 H 46 O 5 Na,m/z 509.3243).
Example 13
Preparation of derivatives 20 and 21:
preparation of derivatives 20 and 21 reference the synthesis of derivatives 10 and 11 at a reaction temperature of 60℃with yields of 40% and 47%, respectively.
Derivative 20: c (C) 28 H 40 O 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝1.9Hz,1H),6.16(d,J＝1.9Hz,1H),5.95(ddt,J＝17.2,10.2,5.0Hz,1H),5.33(dq,J＝17.2,1.9Hz,1H),5.13(dq,J＝10.6,1.7Hz,1H),4.52(t,J＝9.2Hz,1H),4.08(dt,J＝5.0,1.7Hz,2H),4.06(d,J＝2.6Hz,1H),3.17(s,1H),2.81(m,1H),2.69–2.55(m,3H),2.29(m,1H),2.25(t,J＝2.6Hz,1H),2.21(td,J＝10.0,9.4,5.0Hz,1H),1.92(s,4H),1.85(dt,J＝12.5,6.0Hz,2H),1.57(s,3H),1.18(d,J＝7.0Hz,3H),1.15(m,1H),1.06(d,J＝6.8Hz,3H),1.01(d,J＝5.9Hz,3H),0.94(dt,J＝8.6,4.8Hz,1H),0.90(m,1H),0.85(m,1H),0.22(ddd,J＝8.9,5.2,3.6Hz,1H)； 13 CNMR(151MHz,Acetone-d 6 )δ152.3,140.5,136.6,115.8,113.8,113.6,108.9,86.6,84.1,79.3,77.5,71.3,57.2,48.0,46.1,44.4,43.1,34.4,33.6,26.6,23.7,22.1,19.6,19.2,16.1,12.1,9.9,8.9；HRMS(ESI)at m/z 479.2770[M+Na] + (calcd.for C 28 H 40 O 5 Na,m/z 479.2773).
Derivative 21: c (C) 31 H 44 O 5 ,yellow oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝1.8Hz,1H),6.16(d,J＝1.8Hz,1H),5.95(ddt,J＝17.0,10.3,5.0Hz,1H),5.87(ddt,J＝17.0,10.5,4.5Hz,1H),5.33(dt,J＝17.3,1.9Hz,1H),5.27(dt,J＝17.2,2.1Hz,1H),5.12(dq,J＝10.6,1.7Hz,1H),5.02(dq,J＝10.6,1.9Hz,1H),4.57–4.49(m,2H),4.15(ddd,J＝13.3,4.5,2.2Hz,1H),4.09(d,J＝4.8Hz,2H),4.06(d,J＝3.7Hz,1H),2.80(m,1H),2.71(tt,J＝6.7,3.4Hz,1H),2.69–2.62(m,2H),2.58(td,J＝14.8,13.6,5.0Hz,1H),2.34(dt,J＝10.5,2.7Hz,1H),2.24(td,J＝9.6,5.1Hz,1H),1.96(m,1H),1.93(s,3H),1.89–1.81(m,2H),1.55(s,3H),1.22–1.13(m,4H),1.06(d,J＝6.7Hz,4H),1.03(d,J＝1.8Hz,4H),0.94(dt,J＝7.0,3.5Hz,1H),0.24(dd,J＝8.2,4.3Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ152.5,140.5,137.2,136.6,115.8,114.5,113.8,113.6,108.1,89.5,85.8,78.5,77.4,71.4,64.9,57.5,48.6,45.8,44.7,43.1,34.2,28.8,26.66,23.7,22.1,21.6,19.1,16.7,13.1,11.1,9.9；HRMS(ESI)at m/z 519.3081[M+Na] + (calcd.for C 31 H 44 O 5 Na,m/z 519.3086).
Example 14
Preparation of derivative 22:
dissolving colquhounod D (1.0 eq) in CHCl 3 /MeCN/H 2 To a mixed solvent of O (0.37M/0.08M/0.04M), then ruthenium trichloride (0.3 eq) and sodium periodate (30.0 eq) were added in this order. The reaction mixture was stirred at room temperature for 25min. After complete consumption of the starting material by TLC, the reaction was filtered (EA rinsed), the filtrate was collected, the aqueous phase was extracted with EA, and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (PE/EA 3:1-1:1,1:1 with one percent glacial acetic acid added to the eluent) to give derivative 22 in 76% yield.
Derivative 22: c (C) 21 H 30 O 6 ,colorless oil. 1 H NMR(400MHz,Acetone-d 6 )δ4.23(t,J＝2.7Hz,1H),2.49(m,2H),2.37(ddd,J＝16.0,9.4,6.5Hz,1H),2.33(dt,J＝8.1,2.4Hz,1H),2.29(td,J＝8.5,7.6,3.3Hz,1H),2.22(m,1H),2.11(ddd,J＝13.7,9.6,6.8Hz,1H),1.96–1.88(m,2H),1.28(ddd,J＝16.7,7.6,3.3Hz,2H),1.15(d,J＝7.0Hz,3H),1.14–1.09(m,6H),1.04(h,J＝4.3,3.9Hz,4H),0.91(dq,J＝4.7,2.7Hz,2H),0.28(dd,J＝8.6,2.0Hz,1H)； 13 C NMR(101MHz,Acetone-d 6 )δ212.5,175.1,109.1,85.9,82.5,78.4,65.6,54.6,46.0,42.7,39.8,38.3,32.9,29.8,26.1,24.0,19.1,18.9,15.4,12.5,8.9；HRMS(ESI)at m/z 401.1938[M+Na] + (calcd.for C 21 H 30 O 6 Na,m/z 401.1935).
Example 15
Preparation of derivatives 23 and 24:
derivative 22 (1.0 eq), aniline (4.0 eq), DMAP (0.8 eq) and EDCI (4.0 eq) were dissolved in DCM under argon and the reaction stirred at room temperature for 4.5h. After complete consumption of the starting material by TLC, the reaction solution was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel (PE/EA 8:1-1:1) to give derivatives 23 and 24 in 21% and 62% yields, respectively.
Derivative 23: c (C) 21 H 28 O 5 ,colorless crystal. 1 H NMR(600MHz,Acetone-d 6 )δ4.28(t,J＝2.7Hz,1H),3.26(s,1H),2.80(m,1H),2.69(qd,J＝6.9,3.5Hz,1H),2.57(ddd,J＝14.4,6.4,4.1Hz,1H),2.41(ddd,J＝7.7,3.5,2.1Hz,1H),2.36(qd,J＝8.0,2.5Hz,1H),2.30(dt,J＝7.8,2.0Hz,1H),1.94(dd,J＝13.4,8.2Hz,1H),1.92–1.82(m,2H),1.32(ddd,J＝13.6,8.2,3.3Hz,1H),1.24(s,3H),1.16(dd,J＝7.0,6.2Hz,6H),1.12(dt,J＝8.8,4.9Hz,1H),1.05(d,J＝6.0Hz,3H),0.85(m,1H),0.75(dt,J＝8.6,4.7Hz,1H),0.28(ddd,J＝8.9,5.6,4.3Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ209.5,172.8,107.8,91.1,86.2,78.3,58.6,54.2,45.3,42.0,38.9,38.3,32.7,31.6,27.4,23.8,19.2,19.0,15.4,12.7,9.8；HRMS(ESI)at m/z 383.1835[M+Na] + (calcd.for C 21 H 28 O 5 Na,m/z 383.1834).
Derivative 24: c (C) 27 H 35 NO 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ9.17(s,1H),7.67(d,J＝8.0Hz,2H),7.27(t,J＝7.9Hz,2H),7.02(t,J＝7.3Hz,1H),4.21(t,J＝2.6Hz,1H),3.96(s,1H),2.54(d,J＝1.7Hz,1H),2.47(tt,J＝8.4,6.4Hz,2H),2.38(ddd,J＝15.2,8.6,5.6Hz,1H),2.31(dt,J＝8.0,2.6Hz,1H),2.28(m,1H),2.25(m,1H),2.22(d,J＝6.0Hz,1H),1.96(m,1H),1.90(dd,J＝13.3,8.1Hz,1H),1.27(m,1H),1.15(d,J＝7.1Hz,3H),1.13(s,3H),1.11(d,J＝6.9Hz,3H),1.02(m,1H),0.99(d,J＝5.7Hz,3H),0.87(qd,J＝5.1,3.1Hz,2H),0.16(d,J＝9.1Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.6,172.4,140.7,129.5,123.9,120.0,109.0,86.0,82.5,78.4,66.1,54.5,45.9,42.7,39.9,38.3,33.0,32.8,26.3,24.0,19.1,19.0,15.4,12.4,9.2；HRMS(ESI)at m/z 476.2408[M+Na] + (calcd.for C 27 H 35 NO 5 Na,m/z 476.2413).
Example 16
Preparation of derivative 25:
derivative 22 (1.0 eq) was dissolved in dry DMF under argon, then EDCI (3.0 eq), HOBt (2.4 eq), benzyl mercaptan (2.4 eq) and DMAP (0.8 eq) were added in sequence and the reaction stirred at room temperature for 5h. After complete consumption of the starting material by TLC, the reaction was diluted with EA and washed with water. The aqueous phase was extracted again with EA and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (PE/EA 15:1-1:1) to give derivative 25 in 45% yield.
Derivative 25: c (C) 28 H 36 O 5 S,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.34–7.27(m,4H),7.24(m,1H),4.23(t,J＝2.7Hz,1H),4.18–4.09(m,2H),3.68(s,1H),2.72–2.59(m,2H),2.51(qd,J＝6.9,3.4Hz,1H),2.48(d,J＝1.7Hz,1H),2.33(dt,J＝8.1,2.8Hz,1H),2.28(tdd,J＝8.3,6.9,3.2Hz,1H),2.22(d,J＝8.0Hz,1H),2.16(ddd,J＝13.4,10.2,6.1Hz,1H),1.97(ddd,J＝13.4,10.2,5.6Hz,1H),1.91(dd,J＝13.3,8.2Hz,1H),1.28(m,1H),1.15(d,J＝7.0Hz,3H),1.13–1.08(m,6H),1.07–1.00(m,3H),0.95–0.85(m,3H),0.29(ddt,J＝8.2,3.5,2.1Hz,1H)； 13 CNMR(151MHz,Acetone-d 6 )δ212.4,198.8,139.2,129.7,129.5,128.0,109.2,85.8,82.5,78.3,65.4,54.5,46.1,42.7,40.10,40.05,38.3,33.4,32.9,26.1,24.0,19.1,18.8,15.4,12.6,8.8；HRMS(ESI)at m/z 507.2175[M+Na] + (calcd.for C 28 H 36 O 5 SNa,m/z507.2181).
Example 17
Preparation of derivative 26:
preparation of derivative 26 referring to the synthetic method of derivative 24, the yield was 51%.
Derivative 26: c (C) 27 H 33 BrO 6 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.57(d,J＝8.8Hz,2H),7.12(d,J＝8.8Hz,2H),4.25(t,J＝2.7Hz,1H),3.70(s,1H),2.65(dt,J＝9.3,5.9Hz,2H),2.54(s,1H),2.52(dt,J＝10.1,5.0Hz,1H),2.35(m,1H),2.31(ddd,J＝10.3,7.8,3.8Hz,1H),2.24(d,J＝7.7Hz,1H),2.20(m,1H),2.04(m,1H),1.93(dd,J＝13.3,8.1Hz,1H),1.29(ddd,J＝12.6,8.9,3.2Hz,1H),1.16(d,J＝8.8Hz,6H),1.11(d,J＝6.8Hz,3H),1.06(dd,J＝8.8,4.5Hz,1H),1.04(d,J＝5.4Hz,3H),0.92(tq,J＝8.4,4.1Hz,2H),0.29(m,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.5,172.6,151.4,133.2,125.0,119.0,109.3,85.8,82.6,78.4,65.4,54.6,46.1,42.8,39.6,38.4,32.9,30.4,26.2,24.0,19.1,18.8,15.4,12.6,8.9；HRMS(ESI)at m/z 555.1353[M+Na] + (calcd.for C 27 H 33 BrO 6 Na,m/z 555.1358).
Example 18
Preparation of derivative 27
Derivative 22 (1.0 eq), N-methylpiperazine (2.0 eq), HATU (1.2 eq) and DIPEA (2.0 eq) were dissolved in dry DCM under argon and the reaction stirred at room temperature for 4.5h. After complete consumption of the starting material by TLC, the reaction was concentrated under reduced pressure, the residue was purified by column chromatography on silica gel, the impurities were removed by PE/EA1:1 isocratic elution, and derivative 27 was obtained in 39% yield by eluting with 100% MeOH.
Derivative 27: c (C) 26 H 40 N 2 O 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ4.20(s,1H),3.56–3.51(m,2H),3.49(m,2H),2.53–2.48(m,2H),2.45(qd,J＝7.0,3.4Hz,1H),2.37-2.29(m,2H),2.31-2.27(m,5H),2.25(m,1H),2.24–2.18(m,4H),1.89(m,1H),1.82(m,1H),1.26(m,1H),1.14(d,J＝7.0Hz,3H),1.11(s,3H),1.10(d,J＝6.8Hz,3H),1.03(d,J＝5.6Hz,3H),1.03–0.99(m,1H),0.86(qd,J＝7.8,4.1Hz,3H),0.24(d,J＝8.2Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.4,172.0,108.7,85.6,82.3,78.2,66.5,55.9,55.4,54.4,46.2,45.9,45.7,42.5,42.3,39.4,38.2,32.5,28.6,26.4,23.9,19.1,19.0,15.2,12.4,9.1；HRMS(ESI)at m/z 483.2834[M+Na] + (calcd.for C 26 H 40 N 2 O 5 Na,m/z 483.2835).
Example 19
Preparation of derivative 28:
derivative 24 (1.0 eq) was dissolved in dry DMF under the protection of argon, then EDCI (3.0 eq), HOBt (2.4 eq), ethanolamine (2.4 eq) and DMAP (0.8 eq) were added in sequence and the reaction stirred at room temperature overnight. After complete consumption of starting material by TLC, the reaction was diluted with EA and washed with water. The aqueous phase was extracted with EA and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel eluting with PE/EA1:1 isocratically to remove impurities and then DCM/MeOH 10:1 isocratically to give derivative 28 in 52% yield.
Derivative 28: c (C) 23 H 35 NO 6 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.20(s,1H),4.21(t,J＝2.7Hz,1H),4.05(s,1H),3.92(t,J＝5.5Hz,1H),3.56(q,J＝5.6Hz,2H),3.33(dq,J＝13.6,5.7Hz,1H),3.25(dq,J＝13.6,5.6Hz,1H),2.51(d,J＝1.7Hz,1H),2.47(qd,J＝7.0,3.4Hz,1H),2.33–2.23(m,3H),2.21(m,1H),2.19(m,1H),2.13(m,1H),1.93–1.85(m,2H),1.25(m,1H),1.15(d,J＝7.0Hz,3H),1.10–1.08(m,6H),1.04–1.01(m,4H),0.90(m,1H),0.87(m,1H),0.26(ddd,J＝8.5,4.7,3.1Hz,1H).； 13 C NMR(151MHz,Acetone-d 6 )δ212.5,174.1,108.9,85.9,82.4,78.3,65.8,62.1,54.5,45.8,43.1,42.6,40.1,38.2,32.7,31.7,26.3,23.9,19.1,18.9,15.3,12.4,9.0；HRMS(ESI)at m/z 444.2364[M+Na] + (calcd.for C 23 H 35 NO 6 Na,m/z 444.2362).
Example 20
Preparation of derivative 29:
Preparation of derivative 29 referring to the synthetic method of derivative 27, the yield was 42%.
Derivative 29: c (C) 25 H 39 NO 7 ,colorless oil. 1 H NMR(800MHz,Acetone-d 6 )δ4.36(s,1H),4.20(t,J＝2.8Hz,1H),4.09(s,1H),3.75(p,J＝5.3Hz,2H),3.71–3.67(m,2H),3.65(m,1H),3.62–3.53(m,2H),3.39(m,1H),2.63(dt,J＝16.0,7.4Hz,1H),2.52(d,J＝1.7Hz,1H),2.45(qd,J＝6.9,3.3Hz,1H),2.35(dt,J＝16.1,6.7Hz,1H),2.30(dt,J＝8.1,2.8Hz,1H),2.28(q,J＝3.5,3.1Hz,1H),2.26(m,1H),2.20(dt,J＝8.2,2.2Hz,1H),1.89(dd,J＝13.3,8.2Hz,1H),1.85(dt,J＝13.6,6.7Hz,1H),1.26(m,1H),1.14(d,J＝7.0Hz,3H),1.12(s,3H),1.10(d,3H),1.04(d,J＝5.9Hz,3H),1.02(m,1H),0.89(m,1H),0.86(m,1H),0.26(ddd,J＝8.9,5.2,3.7Hz,1H)； 13 C NMR(201M Hz,Acetone-d 6 )δ212.5,175.1,108.8,85.7,82.3,78.4,66.4,61.3,60.9,54.4,52.1,50.5,45.7,42.5,39.6,38.2,32.5,28.9,26.4,23.9,19.1,19.0,15.2,12.1,9.4；HRMS(ESI)at m/z 488.2622[M+Na] + (calcd.for C 25 H 39 NO 7 Na,m/z488.2624).
Example 21
Preparation of derivative 30
Derivative 22 (1.0 eq) was dissolved in DMF under argon, then HATU (1.0 eq), TEA (2.2 eq) and propargylamine (1.2 eq) were added in sequence and the reaction stirred at room temperature for 9h. After the complete consumption of the starting material was monitored by TLC, the reaction solution was diluted with water, extracted with diethyl ether and the organic phases combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (PE/EA 3:1-1:1) to give derivative 30 in 80% yield.
Derivative 30: c (C) 24 H 33 NO 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.43(s,1H),4.21(t,J＝2.8Hz,1H),4.04–3.93(m,2H),3.94(s,1H),2.62(t,J＝2.6Hz,1H),2.50(d,J＝1.7Hz,1H),2.47(m,1H),2.31(dd,J＝6.0,2.6Hz,1H),2.28(m,2H),2.20(dd,J＝9.1,4.2Hz,2H),2.17(m,1H),1.89(ddd,J＝17.1,13.4,6.5Hz,2H),1.25(m,1H),1.15(d,J＝7.0Hz,3H),1.10(dd,J＝6.4,1.5Hz,6H),1.05–1.00(m,4H),0.89(m,2H),0.27(dt,J＝8.7,2.1Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.5,173.2,109.0,86.0,82.5,81.6,78.4,72.0,66.00,54.5,45.9,42.7,40.0,38.3,32.8,31.6,29.1,26.3,24.0,19.2,19.00,15.4,12.4,9.1；HRMS(ESI)at m/z 438.2255[M+Na] + (calcd.for C 24 H 33 NO 5 Na,m/z 438.2256).
Example 22
Preparation of derivative 31
Derivative 24 (1.0 eq) was dissolved in Py under the protection of argon, and DPPA (2.0 eq) was then slowly added dropwise. The reaction mixture was heated to reflux and stirred overnight. After monitoring complete consumption of the starting material by TLC, EA was added to the reaction solution for dilution, followed by washing with 1N HCl and water, drying the organic phase over anhydrous sodium sulfate, filtering, and concentrating the filtrate under reduced pressure. The residue was purified by column chromatography on silica gel (PE/EA 2:1) to give the crude product, which was further purified by semi-preparative HPLC to give derivative 31 in 15% yield.
Derivative 31: c (C) 27 H 34 N 4 O 4 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.72–7.64(m,5H),4.21(s,1H),3.81(d,J＝2.2Hz,1H),3.04–2.92(m,2H),2.51–2.47(m,1H),2.47(s,1H),2.33–2.29(m,2H),2.26(m,1H),2.20(m,1H),2.07(m,1H),1.89(dd,J＝13.4,8.2Hz,1H),1.27(m,1H),1.14(d,J＝7.0Hz,3H),1.11(s,3H),1.09(d,J＝7.0Hz,3H),1.01(d,J＝6.0Hz,3H),0.99(m,1H),0.85(m,1H),0.67(m,1H),0.21(m,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.3,156.6,135.3,131.3,130.9,126.2,109.2,85.7,82.5,78.4,65.5,54.5,46.0,42.7,41.9,38.3,32.8,25.9,24.0,19.9,19.1,18.8,15.4,12.5,9.1；HRMS(ESI)at m/z 501.2477[M+Na] + (calcd.for C 27 H 34 N 4 O 4 Na,m/z 501.2478).
Example 23
Preparation of derivative 32:
derivative 26.0 eq) was dissolved in dry MeCN under argon and DPPA (1.5 eq) and TEA (1.5 eq) were added. The reaction solution was then heated to 45℃and stirred for 5h. After complete consumption of the starting material by TLC, the reaction solution was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel (PE/EA 6:1-1:1) to give derivative 32 in 58% yield.
Derivative 32: c (C) 21 H 30 N 4 O 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ6.85(s,1H),4.24(t,J＝2.6Hz,1H),3.70(s,1H),3.27(tq,J＝10.6,7.6,7.2Hz,2H),2.56–2.51(m,1H),2.51(d,J＝1.8Hz,1H),2.37–2.32(m,1H),2.29(qd,J＝8.0,7.4,2.4Hz,1H),2.22(dd,J＝7.7,2.9Hz,1H),2.02(m,1H),1.92(dd,J＝13.3,8.1Hz,1H),1.85(ddd,J＝13.8,8.6,6.1Hz,1H),1.28(ddd,J＝12.6,8.8,3.3Hz,1H),1.15(t,J＝3.6Hz,6H),1.10(d,J＝6.9Hz,3H),1.06(m,1H),1.04(d,J＝5.6Hz,3H),0.92(m,2H),0.31(d,J＝8.8Hz,1H)； 13 C NMR(151MHz,Acetone)δ212.5,156.4,109.3,85.7,82.6,78.4,65.5,54.6,46.1,44.1,42.8,38.3,38.0,32.9,26.3,24.0,19.1,18.8,15.4,12.5,8.9；HRMS(ESI)at m/z 441.2117[M+Na] + (calcd.for C 21 H 30 N 4 O 5 Na,m/z441.2114).
Example 24
Preparation of derivative 33:
derivative 30 (1.0 eq) was dissolved in dry MeCN under argon and PPh was then added 3 AuNTf 2 (0.2 eq). The reaction mixture was stirred at room temperature for 8.5h. After complete consumption of the starting material by TLC, the reaction solution was concentrated under reduced pressure to remove the solvent, and the residue was purified by silica gel column chromatography (PE/EA 3:1-1:1) to give an intermediate. Then, intermediate (1.0 eq) was dissolved in DCM under the protection of argon, and after NBS (1.5 eq) was added thereto, the reaction solution was stirred at room temperature for 10.0h. After complete consumption of the starting material by TLC, the reaction solution was concentrated under reduced pressure to remove the organic solvent, and the residue was purified by silica gel column chromatography (PE/EA 5:1-1:1) to give derivative 33 in 84% yield.
Derivative 33: c (C) 24 H 32 BrNO 5 ,oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.05(s,1H),4.71(s,2H),4.24(s,1H),2.80–2.75(m,2H)2.55(d,J＝1.6Hz,1H),2.51(m,1H),2.34(m,1H),2.33–2.26(m,2H),2.24(d,J＝6.6Hz,1H),2.10(q J＝14.6Hz,1H),1.93(m,1H),1.29(m,1H),1.18(s,3H),1.16(d,J＝6.9Hz,3H),1.12(d,J＝6.8Hz,3H),1.04(d,J＝5.7Hz,4H),0.91(m,1H),0.82(dt,J＝8.6,4.5Hz,1H),0.27(dt,J＝8.9,4.6Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.4,166.8,148.7,127.0,109.2,85.8,82.5,78.4,65.6,54.6,46.1,42.8,41.4,38.4,32.9,26.1,24.3,24.0,21.7,19.1,18.9,15.4,12.6,9.0；HRMS(ESI)at m/z 516.1358[M+Na] + (calcd.for C 24 H 32 BrNO 5 Na,m/z 516.1362).
Example 25
Preparation of derivative 34:
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derivative 30 (1.0 eq) was dissolved in dry MeCN under argon and PPh was then added 3 AuNTf 2 (0.2 eq). The reaction mixture was stirred at room temperature for 8.5h. After complete consumption of the starting material by TLC, the reaction solution was concentrated under reduced pressure to remove the solvent, and the residue was purified by silica gel column chromatography (PE/EA 3:1-1:1) to give an intermediate.
Method a: under the protection of argon, the intermediate (1.0 eq) was dissolved in DCM and then 3HF.Et was added 3 N1.5 eq) was stirred at room temperature. After the complete consumption of the starting material was monitored by TLC, the reaction solution was concentrated under reduced pressure to remove the organic solvent. The residue was purified by column chromatography on silica gel eluting with PE/EA 1:1 isocratic to remove impurities and then with DCM/MeOH 10:1 isocratic to give derivative 34 in 50% yield.
Method b: the intermediate (1.0 eq) was dissolved in toluene under argon and then HCl in dioxane was added and the reaction was stirred at room temperature. After the complete consumption of the starting material was monitored by TLC, the reaction mixture was quenched by adding aqueous sodium bicarbonate, the aqueous phase was extracted with ethyl acetate, and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography to give derivative 34.
Method c: intermediate (1.0 eq) was dissolved in toluene under the protection of argon, and DBU (0.4 eq) was added to the reaction solution after the temperature was reduced to 0 ℃. The reaction solution was then warmed to room temperature and stirred for 5 hours. After the complete consumption of the starting material was monitored by TLC, the reaction solution was concentrated under reduced pressure to remove the organic solvent. The residue was purified by silica gel column chromatography to give derivative 34.
Derivative 34: c (C) 24 H 35 NO 6 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.29(s,1H),4.21(s,1H),4.10–3.97(m,2H),3.95(s,1H),2.51(d,J＝1.7Hz,1H),2.47(qd,J＝7.0,3.3Hz,1H),2.36(ddd,J＝15.6,9.0,7.0Hz,1H),2.31(dt,J＝8.1,2.8Hz,1H),2.28(m,1H),2.25(ddd,J＝15.0,9.0,5.8Hz,1H),2.21(d,J＝5.6Hz,1H),2.15(m,1H),2.11(s,3H),1.90(ddd,J＝14.0,8.7,5.4Hz,2H),1.25(m,1H),1.15(d,J＝7.0Hz,3H),1.10(d,J＝8.3Hz,6H),1.03(d,J＝3.5Hz,3H),1.02(q,J＝10.9Hz,1H),0.88(m,2H),0.26(d,J＝8.4Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.6,204.4,17 3.8,109.0,86.0,82.5,78.4,65.9,54.6,50.2,45.9,42.7,40.1,38.3,32.8,31.6,27.1,26.3,24.0,19.2,19.0,15.4,12.4,9.1；HRMS(ESI)at m/z 456.2356[M+Na] + (calcd.for C 24 H 35 NO 6 Na,m/z 456.2357).
Example 26
Preparation of derivative 35:
derivative 30 (1.0 eq) and azide reagent (1.0 eq) were dissolved in EtOH/H under argon 2 O (1:1) and then sodium ascorbate (0.3 eq) and CuSO are added 4 ·5H 2 O (0.1 eq) was reacted at room temperature with stirring for 12h. After complete consumption of the starting material by TLC, diluted with EA and water, the aqueous phase extracted with EA and the organic phases combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel eluting with 100% EA to remove impurities and then with DCM/MeOH 10:1 to give derivative 35 in 89% yield.
Derivative 35: c (C) 31 H 40 N 4 O 6 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ8.37(s,1H),7.83(d,J＝8.3Hz,2H),7.64(s,1H),7.57(d,J＝8.1Hz,2H),4.72(d,J＝6.0Hz,2H),4.56(dd,J＝15.2,5.7Hz,1H),4.47(dd,J＝15.2,5.5Hz,1H),4.41(t,J＝5.8Hz,1H),4.21(t,J＝2.6Hz,1H),4.01(s,1H),2.52(d,J＝1.8Hz,1H),2.47(dt,J＝9.5,4.9Hz,1H),2.34(dt,J＝16.0,8.2Hz,1H),2.30(m,1H),2.28(m,1H),2.25(m,1H),2.20(d,J＝6.9Hz,1H),2.17(m,1H),1.93(m,1H),1.89(m,1H),1.26(dq,J＝8.8,4.4,3.2Hz,1H),1.15(d,J＝7.0Hz,3H),1.12–1.07(m,6H),1.02(d,J＝5.6Hz,4H),0.89(m,1H),0.87(m,1H),0.26(dt,J＝8.6,4.2Hz,1H). 13 C NMR(151MHz,Acetone-d 6 )δ212.6,173.6,147.3,144.1,137.0,128.6,121.5,120.9,109.0,86.0,82.5,78.4,65.9,64.0,54.6,45.9,42.7,40.1,38.3,35.6,32.8,31.8,26.4,24.0,19.2,19.0,15.4,12.6,9.1；HRMS(ESI)at m/z 587.2848[M+Na] + (calcd.for C 31 H 40 N 4 O 6 Na,m/z 587.2846).
Example 27
Preparation of derivative 36:
preparation of derivative 36 referring to the synthetic method of derivative 1, the reaction temperature was 0 ℃ and the yield was 35%.
Derivative 36: c (C) 21 H 34 O 5 ,white powder. 1 H NMR(600MHz,Acetone-d 6 )δ4.92(td,J＝9.3,5.8Hz,1H),4.04(t,J＝2.5Hz,1H),3.65(d,J＝5.6Hz,1H),3.49(ddt,J＝15.6,10.2,5.8Hz,2H),3.37(t,J＝5.4Hz,1H),2.96(s,1H),2.86(m,1H),2.47(d,J＝9.8Hz,1H),2.26(qd,J＝7.1,2.6Hz,1H),2.21(dt,J＝10.4,2.7Hz,1H),2.02(m,1H),1.83(dd,J＝12.6,7.6Hz,1H),1.65–1.56(m,4H),1.56(s,3H),1.17(d,J＝7.1Hz,3H),1.13(ddd,J＝13.0,10.6,2.6Hz,1H),1.04(d,J＝6.8Hz,3H),1.00(d,J＝6.0Hz,3H),0.93(dt,J＝8.5,4.9Hz,1H),0.88(m,1H),0.80(ddd,J＝8.5,5.0,3.6Hz,1H),0.18(ddd,J＝8.8,5.3,3.6Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ108.8,87.4,84.0,79.5,69.4,63.4,59.1,48.2,47.2,44.6,43.1,34.0,33.7,29.4,27.2,24.2,19.8,19.2,16.1,12.0,9.0；HRMS(ESI)at m/z 389.2299[M+Na] + (calcd.for C 21 H 34 O 5 Na,m/z 389.2298).
Example 28
Preparation of derivatives 37 and 38:
under the protection of argon, colquhouoid D (1.0 eq) was dissolved in dry DCM, then m-CPBA (2.0 eq) and NaHCO were added sequentially 3 (2.0 eq) and the reaction mixture was stirred at room temperature for 4h. After complete consumption of the starting material by TLC, the reaction was quenched by addition of saturated sodium bicarbonate solution, the aqueous phase extracted with DCM and the organic phases combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel eluting with PE/EA1:1 isocratically to remove impurities and DCM/MeOH 10:1 isocratically to give derivatives 37 and 38 in 61% yield.
Derivative 37: c (C) 25 H 34 O 7 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ6.22(s,1H),5.79(q,J＝1.7Hz,1H),4.23(d,J＝3.5Hz,1H),3.65(s,1H),2.50(qd,J＝7.0,4.7,3.5Hz,1H),2.45(d,J＝10.1Hz,1H),2.33(dq,J＝9.3,3.7,3.2Hz,1H),2.29(tt,J＝8.2,3.8Hz,1H),2.21(d,J＝7.1Hz,1H),2.09(t,J＝2.1Hz,1H),2.03(s,3H),1.91(dd,J＝13.2,8.6Hz,1H),1.84(td,J＝14.4,13.1,3.2Hz,,1H),1.61(m,2H),1.27(m,1H),1.15(d,J＝7.0Hz,3H),1.13(s,3H),1.09(d,J＝5.4Hz,3H),1.03(dd,J＝5.8,2.9Hz,4H),0.94–0.89(m,2H),0.28(d,J＝6.3Hz,1H). 13 C NMR(151MHz,Acetone-d 6 )δ212.5,170.7,168.0,118.7,109.1,109.0,86.0,82.5,78.3,65.3,54.6,46.1,42.8,38.3,33.0,31.9,26.1,24.0,19.1,18.8,15.4,12.6,8.8；HRMS(ESI)at m/z 469.2199[M+Na] + (calcd.for C 25 H 34 O 7 Na,m/z 469.2197).
Derivative 38: c (C) 25 H 34 O 7 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ6.24(s,1H),5.79(q,J＝1.7Hz,1H),4.23(d,J＝3.5Hz,1H),3.60(s,1H),2.50(qd,J＝7.0,4.7,3.5Hz,1H),2.45(d,J＝10.1Hz,1H),2.33(dq,J＝9.3,3.7,3.2Hz,1H),2.29(tt,J＝8.2,3.8Hz,1H),2.21(d,J＝7.1Hz,1H),2.09(t,J＝2.1Hz,1H),2.03(s,3H),1.91(dd,J＝13.2,8.6Hz,1H),1.84(m,1H),1.76(td,J＝12.5,3.8Hz,1H),1.56(td,J＝12.9,3.8Hz m,1H),1.27(m,1H),1.15(d,J＝7.0Hz,3H),1.13(s,3H),1.09(d,J＝5.4Hz,3H),1.03(dd,J＝5.8,2.9Hz,4H),0.94–0.89(m,2H),0.28(d,J＝6.3Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.6,170.7,168.0,118.7,109.1,109.0,86.1,82.5,78.3,65.1,54.6,46.1,42.8,38.3,33.0,32.1,26.2,24.0,19.1,18.8,15.4,12.6,12.5,8.8；HRMS(ESI)at m/z 469.2199[M+Na] + (calcd.for C 25 H 34 O 7 Na,m/z 469.2197).
Example 29
Preparation of derivatives 39 and 40:
under the protection of argon, colquhounod D (1.0 eq) was dissolved in THF, then 0.5mL of 2M HCl was slowly added dropwise to the reaction solution. After the completion of the dropwise addition, the reaction mixture was stirred at 45℃for 23 hours. After complete consumption of the starting material by TLC, the reaction was cooled to room temperature and quenched by slow dropwise addition of saturated sodium bicarbonate solution. The aqueous phase was extracted with EA and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the organic solvent. The residue was purified by column chromatography on silica gel (PE/EA 8:1-2:1) to give crude products, which were purified by semi-preparative HPLC to give derivatives 39 and 40 in 16% and 27% yields, respectively.
Derivative 39: c (C) 25 H 35 ClO 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝1.8Hz,1H),6.16(d,J＝1.9Hz,1H),4.19(m,1H),4.05(s,1H),4.03(s,1H),2.70–2.58(m,2H),2.57(d,J＝1.7Hz,1H),2.37(qd,J＝6.9,3.3Hz,1H),2.35–2.29(m,2H),2.24(d,J＝7.9Hz,1H),2.06(s,1H),2.04–1.99(m,2H),1.94(s,3H),1.99–1.88(m,3H),1.82(ddd,J＝13.9,9.8,7.1Hz,1H),1.52(d,J＝6.5Hz,3H),1.28(ddd,J＝12.6,8.9,3.2Hz,1H),1.22(s,3H),1.16(d,J＝7.0Hz,3H),1.11(d,J＝6.9Hz,3H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.6,151.8,140.6,114.2 113.7,110.0,86.5,82.0,78.3,65.2,60.8,54.6,45.9,43.6,42.7,38.3,34.7,32.9,26.11,26.09,25.5,24.0,22.13,15.4,10.0；HRMS(ESI)at m/z 473.2062[M+Na] + (calcd.for C 25 H 35 ClO 5 Na,m/z 473.2065).
Derivative 40: c (C) 25 H 35 ClO 5 ,colorless oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝1.9Hz,1H),6.16(d,J＝1.9Hz,1H),4.07(t,J＝2.5Hz,2H),3.86(dd,J＝10.4,3.5Hz,1H),3.59(dd,J＝10.2,7.0Hz,1H),2.70(m,1H),2.62(m,1H),2.54(d,J＝2.0Hz,1H),2.36(tt,J＝6.6,3.0Hz,1H),2.32(m,2H),2.28(m,1H),2.24(m,1H),2.09(m,1H),1.97–1.93(m,3H),1.94(s,3H),1.83(dd,J＝14.8,4.6Hz,1H),1.26(m,1H),1.23(d,J＝2.9Hz,3H),1.16(d,J＝6.9Hz,3H),1.10(td,J＝4.6,2.3Hz,6H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.6,151.8,140.6,114.2,113.7,110.1,86.7,82.3,78.5,65.2,54.6,52.9,45.9,43.8,42.7,38.3,32.8,32.0,31.4,26.1,24.0,22.0,19.7,15.5,9.9；HRMS(ESI)at m/z 473.2063[M+Na] + (calcd.for C 25 H 35 ClO 5 Na,m/z 473.2065).
Example 30
Preparation of derivative 41:
preparation of derivative 41 referring to the synthetic methods of derivatives 10 and 11, without purification by semi-preparative HPLC, the yield was 68%.
Derivative 41: c (C) 26 H 36 O 5 ,yellow oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝1.8Hz,1H),6.16(d,J＝1.8Hz,1H),4.25(t,J＝2.6Hz,1H),3.51(s,3H),2.97(qd,J＝6.8,3.7Hz,1H),2.63–2.49(m,2H),2.44(d,J＝1.7Hz,1H),2.43(dt,J＝3.7,2.0Hz,1H),2.28(dq,J＝10.8,8.1,7.2Hz,1H),2.24(m,1H),1.99–1.87(m,6H),1.29(ddd,J＝13.3,8.8,3.1Hz,1H),1.21(dd,J＝8.6,4.3Hz,1H),1.16(d,J＝6.9Hz,3H),1.14(s,3H),1.09–1.04(m,7H),0.97(dq,J＝8.7,4.3Hz,1H),0.39(m,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.2,151.8,140.6,114.1,113.7,108.4,87.1,85.7,77.7,65.7,54.9,51.7,46.4,43.2,42.7,38.1,27.0,26.0,24.0,22.0,20.6,19.0,15.9,13.5,11.4,9.9；HRMS(ESI)at m/z 451.2452[M+Na] + (calcd.for C 26 H 36 O 5 Na,m/z 451.2455).
Example 31
Preparation of derivative 42:
preparation of derivative 42 referring to the synthetic methods of derivatives 10 and 11, the reaction temperature was 60 ℃, without purification by semi-preparative HPLC, with a yield of 70%.
Derivative 42: c (C) 28 H 38 O 5 ,yellow oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝2.0Hz,1H),6.16(d,J＝2.0Hz,1H),5.88(ddt,J＝16.9,10.0,4.6Hz,1H),5.27(dt,J＝17.2,2.0Hz,1H),5.04(dt,J＝10.6,2.0Hz,1H),4.60(ddt,J＝13.0,4.1,1.9Hz,1H),4.26(m,1H),4.21(ddd,J＝13.0,5.0,1.8Hz,1H),2.98(qd,J＝6.8,3.7Hz,1H),2.65–2.53(m,2H),2.53(d,J＝1.8Hz,1H),2.43(ddd,J＝7.9,3.8,2.0Hz,1H),2.30(m,1H),2.26(d,J＝8.5Hz,1H),2.05–1.90(m,2H),1.93(s,4H),1.30(ddd,J＝12.8,8.7,3.2Hz,1H),1.23(dq,J＝8.6,4.7,3.4Hz,1H),1.16(t,J＝3.5Hz,6H),1.13–1.02(m,7H),0.99(m,1H),0.34(dt,J＝8.3,4.0Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.1,151.8,140.6,136.7,114.8,114.1,113.6,108.4,87.4,85.8,77.7,65.8,64.7,54.9,46.5,43.4,42.7,38.1,27.6,26.0,24.0,22.0,20.8,19.0,15.9,13.4,11.3,9.9；HRMS(ESI)at m/z 477.2615[M+Na] + (calcd.for C 28 H 38 O 5 Na,m/z 477.2617).
Example 32
Preparation of derivative 43:
preparation of derivative 43 referring to the synthetic method of derivatives 10 and 11, the reaction temperature was 60 ℃, without purification by semi-preparative HPLC, with a yield of 80%.
Derivative 43: c (C) 28 H 36 O 5 ,yellow oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.26(d,J＝1.9Hz,1H),6.16(d,J＝1.9Hz,1H),4.67(dd,J＝15.0,2.4Hz,1H),4.44(dd,J＝15.1,2.4Hz,1H),4.26(t,J＝2.6Hz,1H),2.97(qd,J＝6.9,3.7Hz,1H),2.88(t,J＝2.4Hz,1H),2.65–2.52(m,2H),2.50(d,J＝1.7Hz,1H),2.45(ddd,J＝8.1,3.7,2.1Hz,1H),2.29(d,J＝7.2Hz,1H),2.27(d,J＝8.2Hz,1H),2.01(m,1H),1.95(s,3H),1.94(m,1H),1.92(m,1H),1.30(m,1H),1.24(m,1H),1.16(d,J＝7.7Hz,6H),1.13(d,J＝6.8Hz,3H),1.08–1.02(m,5H),0.43(dd,J＝8.1,3.6Hz,1H). 13 C NMR(151MHz,Acetone-d 6 )δ211.8,151.8,140.6,114.2,113.7,108.2,88.1,85.9,81.8,77.7,74.9,65.6,54.8,52.6,46.5,43.3,42.6,38.1,27.4,26.0,23.9,22.0,20.4,19.0,16.0,13.4,11.3,10.0；HRMS(ESI)at m/z 475.2461[M+Na] + (calcd.for C 28 H 36 O 5 Na,m/z 475.2460).
Example 33
Preparation of derivative 44:
preparation of derivative 44 referring to the synthetic method of derivatives 10 and 11, the reaction temperature was 60 ℃, without purification by semi-preparative HPLC, with a yield of 56%.
Derivative 44: c (C) 32 H 40 O 5 ,yellow oil. 1 H NMR(600MHz,Acetone-d 6 )δ7.37(d,J＝7.1Hz,2H),7.29(d,J＝7.5Hz,2H),7.28(d,J＝1.6Hz,1H),7.22(t,J＝7.2Hz,1H),6.16(d,J＝1.9Hz,1H),5.21(d,J＝11.5Hz,1H),4.77(d,J＝11.5Hz,1H),4.31(t,J＝2.7Hz,1H),3.13(qd,J＝6.7,3.9Hz,1H),2.61(m,1H),2.60(d,J＝1.6Hz,1H),2.49(ddd,J＝8.1,3.8,2.2Hz,1H),2.33(m,1H),2.28(m,1H),2.10(m,1H),1.99(m,1H),1.96(m,1H),1.88(s,3H),1.37–1.26(m,2H),1.20–1.16(m,6H),1.12(d,J＝6.8Hz,3H),1.08(s,4H),1.00(s,1H),0.36(dt,J＝8.1,3.9Hz,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ212.1,151.8,140.7,140.6,129.0,127.8,127.6,114.1,113.6,108.4,87.7,85.9,77.7,65.9,65.7,55.0,46.5,43.5,42.7,38.1,27.6,26.0,24.0,21.9,21.0,19.0,16.0,13.6,11.5,9.9,HRMS(ESI)at m/z 527.2771[M+Na] + (calcd.for C 32 H 40 O 5 Na,m/z 527.2773).
Example 34
Preparation of derivatives 45 and 46:
preparation of derivatives 45 and 46 referring to the synthetic method of derivative 1 and derivatives 8 and 9, compounds 45 and 46 were obtained in 61% and 10% yields, respectively.
Derivative 45: c (C) 32 H 40 N 2 O 5 S,yellow oil. 1 H NMR(400MHz,Acetone-d 6 )δ8.38(s,1H),7.76(s,1H),7.19(d,J＝1.9Hz,1H),7.08(s,1H),6.59(t,J＝9.5Hz,1H),6.12(d,J＝1.9Hz,1H),4.66(dd,J＝15.1,2.5Hz,1H),4.42(dd,J＝15.1,2.5Hz,1H),4.19(s,1H),3.08(d,J＝10.1Hz,1H),2.97(m,1H),2.90(qd,J＝6.77,2.7Hz,1H),2.87(t,J＝2.4Hz,1H),2.57–2.44(m,4H),2.00(m,1H),1.96(m,1H),1.88(s,3H),1.80(ddd,J＝13.2,11.1,6.0Hz,1H),1.48(s,3H),1.33(d,J＝6.9Hz,3H),1.30(dd,J＝10.3,2.5Hz,1H),1.16(dt,J＝6.7,3.6Hz,1H),1.09(d,J＝6.8Hz,3H),1.07(s,4H),1.04(m,1H),0.39(m,1H)； 13 C NMR(151MHz,Acetone-d 6 )δ185.1,151.8,140.5,137.9,132.2,119.0,114.1,113.7,108.2,90.7,85.8,82.6,81.9,78.5,74.8,56.2,53.0,48.7,45.1,44.0,42.8,35.7,28.0,27.0,23.2,22.0,21.1,19.0,16.6,13.3,11.5,9.9；HRMS(ESI)at m/z 587.2549[M+Na] + (calcd.for C 32 H 40 N 2 O 5 SNa,m/z587.2556).
Derivative 46: c (C) 32 H 40 N 2 O 6 ,yellow oil. 1 H NMR(400MHz,Acetone-d 6 )δ8.17(s,1H),7.55(s,1H),7.20(d,J＝1.9Hz,1H),7.08(d,J＝1.6Hz,1H),6.13(d,J＝1.8Hz,1H),6.06(t,J＝9.1Hz,1H),4.65(dd,J＝15.1,2.5Hz,1H),4.41(dd,J＝15.1,2.5Hz,1H),4.16(s,1H),2.97–2.88(m,2H),2.90–2.84(m,2H),2.56–2.46(m,4H),1.97(m,1H),1.93(t,J＝5.3Hz,1H),1.89(s,3H),1.84(m,1H),1.54(s,3H),1.36–1.22(m,4H),1.14(m,1H),1.07(d,J＝6.5Hz,7H),1.02(d,J＝5.6Hz,1H),0.38(m,1H)； 13 C NMR(151MHz,Acetone)δ151.9,149.2,140.5,137.9,131.8,118.2,114.1,113.7,108.2,90.6,86.0,82.0,78.5,77.1,74.7,56.9,53.0,48.7,45.3,44.1,42.8,35.2,28.1,27.0,23.2,22.0,21.1,19.0,16.6,13.3,11.5,9.9；HRMS(ESI)at m/z 571.2777[M+Na] + (calcd.for C 32 H 40 N 2 O 6 Na,m/z 571.2779).
Example 35
Preparation of derivative 47:
preparation of derivative 47 referring to the synthetic method of derivative 35, derivative 47 was obtained in 75% yield.
Derivative 47: c (C) 46 H 64 N 8 O 8 S 2 ,white powder. 1 H NMR(400MHz,Acetone-d 6 )δ8.37(s,1H),7.81(s,1H),7.75(t,J＝1.5Hz,1H),7.23(d,J＝1.9Hz,1H),7.08(s,1H),6.96(s,1H),6.60(t,J＝9.6Hz,1H),6.13(d,J＝1.9Hz,1H),5.83(s,1H),5.64(s,1H),5.24(d,J＝11.4Hz,1H),4.85(d,J＝11.4Hz,1H),4.55(t,J＝5.2Hz,2H),4.49(t,J＝6.4Hz,1H),4.32(ddd,J＝7.5,4.6,1.8Hz,1H),4.25(s,1H),3.85(t,J＝5.2Hz,2H),3.48(t,J＝5.6Hz,2H),3.33-3.24(m,2H),3.20(td,J＝7.2,4.3Hz,1H),3.09(d,J＝10.1Hz,1H),3.04(dd,J＝6.8,3.1Hz,1H),2.99(m,1H),2.93(dd,J＝12.5,5.0Hz,1H),2.69(d,J＝12.5Hz,1H),2.58(m,1H),2.55–2.42(m,3H),2.16(t,J＝7.49Hz,2H),2.02-1.95(m,2H),1.81(s,3H),1.79-1.73(m,2H),1.68–1.53(m,3H),1.49(s,3H),1.47 -1.41(m,2H),1.37(d,J＝6.8Hz,3H),1.33(t,J＝2.4Hz,1H),1.30(m,1H),1.10(d,J＝7.2Hz,7H),1.01(m,1H),0.37(m,1H)； 13 CNMR(151MHz,Acetone-d 6 )δ185.1,173.1,163.5,151.9,146.9,140.7,137.9,132.2,123.9,119.0,114.1,113.7,108.3,90.5,85.7,82.7,78.5,70.4,69.9,62.4,60.8,59.3,56.6,56.5,50.7,48.7,45.2,44.1,42.9,41.1,39.6,36.3,35.7,29.2,29.2,28.2,26.9,26.5,23.2,21.2,21.52,19.1,16.7,13.6,11.6,9.9；HRMS(ESI)at m/z 943.4182[M+Na] + (calcd.for C 46 H 64 N 8 O 8 S 2 Na,m/z 943.4186).
Example 36
The compounds 1-47 of examples 1-35 were tested for mouse T cell immunosuppressive activity.
Reagent purchase: PBS, RIPM1640, fetal bovine serum purchased from Biological Industries company; CD3 mAb, CD28 mAb, IFN-gamma detection kit was purchased from BD Bioscience; nylon wool is purchased from PolyScience corporation; CCK8 reagent was purchased from melem corporation.
Preparing a sample to be tested: compounds 1-47 of examples 1-35 were dissolved in DMSO to prepare a 20mM stock solution.
The specific implementation method comprises the following steps: spleen was aseptically isolated from 6-8 week old female mice, ground, filtered through a 200 mesh screen, T lymphocytes were isolated using the Ni Long Maozhu method, the obtained T lymphocytes (4X 105/well) were inoculated into CD3 (5. Mu.g/ml) -coated 96-well plates, each of which was added with the formulated 20, 10, 5, 2.5. Mu.M compound, CD28 (2. Mu.g/ml) mab and IL-2 (40 ng/ml), and the cells were cultured in an incubator at 37℃with 5% CO 2. After 48h, cell supernatants were collected and assayed for secretion of inflammatory factors IFN-gamma by Enzyme-Linked immunosorbent Assay (ELISA) using 10. Mu.L CCK-8 per well and incubated in an incubator for 4 hours with an ELISA reader for OD450 absorbance. The experiments were also performed in the unstimulated blank, DMSO negative control, cyclosporine a (CsA) positive drug group. The inhibitory activity of the compounds on T lymphocyte proliferation and inflammatory factor IFN-gamma was calculated and analyzed for half inhibition concentration IC using GraphPad Prism 8.0 50 The results are shown in Table 1.
TABLE 1 inhibitory Activity of Compounds 1-47 on CD3, CD28 for stimulating proliferation of mouse T lymphocytes and inflammatory factor IFN-gamma
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Watch with a watch1 it can be seen that the in vitro immunosuppressive activity screening of compounds 1-47 on mouse T cells found 28 compounds with enhanced IFN-gamma secretion inhibitory activity in mouse T cells compared to the prototype compound colquhounod D. Wherein 19 structure-modified diterpenoid compounds can obviously inhibit IFN-gamma secretion in the T lymphocytes of mice stimulated by CD3/CD28, and the inhibition rate is more than 50% at 20 mu M. The inhibition rate of the compounds 8 and 32 on the secretion of IFN-gamma by the activated T cells is more than 97 percent. IC for inhibiting secretion of IFN-gamma by T cells by Compounds 8 and 32 50 1.95. Mu.M, 1.75. Mu.M. And the compound does not show obvious toxicity at the dosage, which proves that the compound has stronger research value of immunosuppressive activity.
Example 37
Compounds 1-47 were tested for mouse macrophage BMDM immunosuppressive activity.
Reagent purchase: IL-1. Beta. Detection kit was purchased from Beijing daceae, biotechnology Co., ltd, M-CSF kit was purchased from Peproteck, DMEM medium and fetal bovine serum were purchased from Biological Industries.
Preparing a sample to be tested: the 47 compounds of examples 1 to 33 were dissolved in DMSO to prepare 20mM stock solutions.
The specific method comprises the following steps: mouse primary macrophage BMDM was taken from 8-12 week old C57BL/6 mouse bone marrow. The tibia and femur were first washed with penicillin-streptomycin-containing PBS, centrifuged at 1500rpm for 5 min, the cells resuspended in DMEM medium supplemented with 10% FBS and 50ng/mL M-CSF and plated in 96-well plates to induce differentiation. After 7 days of differentiation, 100ng/mL LPS was added for 3 hours, then the prepared compound was added for 6 hours, and 5mM ATP was added for 30 minutes, and the supernatant was collected. Enzyme-Linked immunosorbent Assay (ELISA) detects IL-1. Beta. Secretion in the supernatant. mu.L of CCK-8 reagent is added into each cell, and OD is measured after incubation for 4 hours at 37 DEG C 450 To evaluate the proliferation of cells. The experiments were performed simultaneously with DMSO negative control, non-stimulated and blank control. The inhibition activity of the compounds on BMDM cell proliferation and IL-1β was calculated and half inhibition concentration IC was analyzed using GraphPad Prism software 50 The results are shown in Table 2
TABLE 2 inhibitory Activity of Compounds 1-47 on LPS+ATP-stimulated proliferation of mouse BMDM cells and cytokine IL-1β production
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As can be seen from Table 2, the in vitro immunosuppressive activity screening of the BMDM cells of the mice was performed on the compounds 1-47, and found that 37 compounds with enhanced secretion inhibition of inflammatory factor IL-1β secreted by BMDM cells stimulated by LPS+ATP of the mice compared with the prototype natural product colquhouoid D, 15 compounds with significantly inhibited secretion of inflammatory factor IL-1β, the inhibition rate at 10. Mu.M was between 50% and 91%, the inhibition rate of the compound 8 on the secretion of activated BMDM cells IL-1β was 91.01%, IC 50 0.253. Mu.M, and showed no significant toxicity. The compounds have outstanding immunosuppressive activity.
Example 38
Test of compound 8 in example 5 for DSS-induced protection against colitis in mice.
Reagent purchase: IL-1. Beta. Detection kit was purchased from Beijing daceae, biotechnology Inc., DSS was purchased from MP Biomedicals, anti-caspase 1-anti-ibody was purchased from Abcam, anti-clean-caspase 1-anti-ibody was purchased from Adipogen, anti-actin-anti-ibody was purchased from Abcam, anti-F4/80-anti was purchased from Zenbio, PBS was purchased from Biological Industries.
Preparing a sample to be tested: compound 8 of example 5 was dissolved in PBS containing 3% dmso and 3% cremophor EL for use.
The specific method comprises the following steps: DSS-induced colitis model was established by giving C57BL/6 mice free drinking for 7 days with 2.5% (w/v) aqueous DSS and then free drinking for 3 days with plain water. Mice were randomly assigned to 5 groups, normal, DSS+Vehicle, DSS+8 (10 mg/kg), DSS+8 (30 mg/kg), DSS+5-ASA (200 mg/kg), 8 groups each. Body weight and Disease Activity Index (DAI), including stool consistency and bleeding, were monitored daily. Mice were sacrificed on day 10, colon tissues were collected, mice were assessed for colon inflammation, including body weight, DAI score, colon length changes, histopathology and scoring thereof, and inflammatory changes in colon tissues were further detected.
As can be seen from FIG. 2, intraperitoneal injection of Compound 8 (30 mg/kg) and oral administration of 5-ASA (200 mg/kg) significantly improved DSS-induced colitis in mice, manifested by weight loss, decreased Disease Activity Index (DAI), and shortened colon (a-d in FIG. 2). Histological analysis showed that compound 8 treatment significantly reduced pathological phenotypes such as inflammatory cell infiltration, mucosal erosion, crypt abscess, and goblet cell loss compared to DSS group (e in fig. 2). In addition, compound 8 significantly reduced infiltration of F4/80+ macrophages (F in fig. 2). Finally, compound 8 also significantly inhibited IL-1β secretion and clear-caspase-1 levels in colon tissue (g-h in FIG. 2), indicating that compound 8 alleviates DSS-induced colitis by inhibiting activation of NLRP3 inflammatory bodies in vivo. The results show that the compound 8 has remarkable immunosuppressive activity in vitro and in vivo and can be used as a medicament.
Formulation examples
In the following preparation examples, conventional reagents are selected and preparation is carried out according to the conventional method, and the application examples only show that the colquhouoid D derivative of the labdane is prepared into different preparations, and specific reagents and operations are not particularly limited.
1. Dissolving any one or any combination of the colquhounod D derivatives 1-47 with DMSO, adding water for injection according to a conventional method, fine filtering, packaging and sterilizing to obtain injection, wherein the concentration of the injection is 0.5-5 mg/mL.
2. Dissolving one or any combination of the colquhounod D derivatives 1-47 in DMSO, dissolving in sterile injectable water, stirring to dissolve, filtering with sterile suction filter funnel, sterile fine filtering, packaging in ampoule, freeze drying at low temperature, and sealing under sterile condition to obtain powder for injection.
3. Adding excipient into one or any combination of the colquhounod D derivatives 1-47 at a mass ratio of 5:1 with excipient, and making into powder.
4. Adding excipient into one or any combination of the colquhounod D derivatives 1-47 at a mass ratio of 5:1 with excipient, granulating, and tabletting.
5. Preparing the colquhounod D derivative 1-47 or their combination into oral liquid by conventional method.
6. Adding excipient into one or any combination of the colquhounod D derivatives 1-47 at a ratio of 5:1 with excipient, and making into capsule.
7. Adding excipient into one or any combination of the colquhounod D derivatives 1-47 at a mass ratio of 5:1 with excipient, and making into granule.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. A colquhouund D derivative of a labdane sesterterpene shown in the following structural formula (I),
in the general formula (I), R 1 Substituents include, but are not limited to, hydrogen, methyl, n-propyl, n-butyl, n-pentyl, allyl, propargyl, benzoyl, acetyl,
R 2 Substituents include, but are not limited to, carboxyl, hydroxymethyl,
R 3 And R is R 4 Substituents include, but are not limited to, substituted or unsubstituted cyclopropane, substituted or unsubstituted n-butyl, substituted or unsubstituted isobutyl, hydrogen atom;
R 5 substituents include, but are not limited to, hydrogen, methyl, allyl, propargyl, benzyl, biotin;
R 6 substituents include, but are not limited to, hydroxy.
2. The derivative of the colquhouund D, which is a colquhoid of the formula (i), according to claim 1, wherein the derivative is any one of the following compounds:
3. a process for the preparation of the colquhounod D derivative 1-21, which is a colquhounod, as claimed in claim 2, characterized in that it comprises the following steps:
(1) Dissolving a compound colquhounod D shown in a structural formula (II) in an organic solvent N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol;
(2) Adding 1.5M diisobutyl aluminum hydride in inert gas atmosphere, stirring at 0-60 ℃ to react until TLC shows complete reaction, quenching the reaction liquid with potassium sodium tartrate, extracting, drying, concentrating in vacuum, and purifying to obtain a colquhouoid D derivative 1-4;
(3) Dissolving the colquhounod D derivative 1 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol;
(4) Under inert gas atmosphere, adding acylating reagent acetic anhydride, benzoic anhydride, acetyl chloride, benzoyl chloride, DMAP or triethylamine, heating to 25-50 ℃, stirring until TLC shows complete reaction, vacuum concentrating and purifying the reaction liquid to obtain a colquhounod D derivative 5-6;
(5) Dissolving the colquhounod D derivative 1 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol;
(6) Adding N, N-thiocarbonyldiimidazole and DMAP under inert gas atmosphere, heating to 60-80 ℃, stirring until TLC shows that the reaction is complete, and vacuum concentrating and purifying the reaction liquid to obtain a colquhounod D derivative 8-9;
(7) Dissolving the colquhounod D derivative 1 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol;
(8) Adding sodium hydride, different substituted isothiocyanates or halogenated hydrocarbon methyl isothiocyanates, phenyl isothiocyanates, pyridine isothiocyanates, methyl iodide, iodopropane, iodobutane or iodopentane in an inert gas atmosphere, heating to 25-60 ℃, stirring for reaction until TLC shows complete reaction, filtering the reaction solution, collecting filtrate, concentrating in vacuum, and purifying to obtain a colquhouoid D derivative 10-21;
(9) Dissolving the colquhounod D derivative 6 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol;
(10) Adding phosphorus pentasulfide and sodium carbonate under inert gas atmosphere, heating to 40-80 ℃, stirring until TLC shows that the reaction is complete, and vacuum concentrating and purifying the reaction liquid to obtain a colquhounod D derivative 7;
4. a process for the preparation of the colquhounod D derivative 22-38, which is a colquhounod, as claimed in claim 2, characterized in that it comprises the following steps:
(1) Dissolving the compound colquhounod D in organic solvent N, N-dimethylformamide, tetrahydrofuran,
Dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol and water;
(2) Adding ruthenium trichloride and sodium periodate, stirring at room temperature for reaction until TLC shows that the reaction is complete, diluting the reaction liquid with organic solvent ethyl acetate or dichloromethane, filtering, washing with brine, drying, concentrating in vacuum and purifying to obtain a colquhounod D derivative 22;
(3) Dissolving the compound colquhounod D in organic solvent N, N-dimethylformamide, tetrahydrofuran,
Dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol;
(4) Adding m-chloroperoxybenzoic acid and sodium bicarbonate in an inert gas atmosphere, stirring at room temperature to react until TLC shows that the reaction is complete, quenching the reaction liquid with sodium bicarbonate solution, extracting, drying, concentrating in vacuum, and purifying to obtain colquhounod D derivatives 37 and 38;
(5) Dissolving the colquhounod D derivative 22 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol;
(6) Under inert gas atmosphere, adding organic amine aniline, ethanolamine, propargylamine, N-methylpiperazine and diethanolamine or p-bromophenol or benzyl mercaptan, condensing agent DMAP, DCC, EDCI, HOBt, HATU,
DIPEA, stirring at room temperature until TLC shows that the reaction is complete, and concentrating and purifying the reaction liquid in vacuum to obtain the colquhounod D derivative 23-30;
(7) Dissolving the colquhounod D derivative 22 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol;
(8) Adding DPPA and triethylamine in inert gas atmosphere, stirring at 45 ℃ for reaction until TLC shows that the reaction is complete, and vacuum concentrating and purifying the reaction liquid to obtain a colquhouoid D derivative 32;
(9) Dissolving the colquhounod D derivative 23 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol;
(10) Adding 1.5M diisobutyl aluminum hydride in inert gas atmosphere, stirring at 0 ℃ to react until TLC shows complete reaction, quenching the reaction liquid with potassium sodium tartrate, extracting, drying, concentrating in vacuum, and purifying to obtain a colquhouoid D derivative 36;
(11) Dissolving the colquhounod D derivative 24 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine, or ethanol;
(12) Under the inert gas atmosphere, DPPA is added, heated and refluxed to 100-130 ℃, stirred until TLC shows that the reaction is complete, and the reaction liquid is concentrated and purified in vacuum to obtain the colquhouoid D derivative 31;
(13) Dissolving the colquhounod D derivative 30 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol;
(14) Under the inert gas atmosphere, PPh is added 3 AuNTf 2 Stirring at room temperature until TLC shows complete reaction, vacuum concentrating and purifying the reaction liquid to obtain an intermediate, dissolving the intermediate in an organic solvent, adding N-bromosuccinimide, triethylamine trihydrofluoride or DBU into the intermediate, stirring at room temperature until TLC monitors complete reaction, vacuum concentrating and purifying the reaction liquid to obtain the colquhounod D derivative 33-34;
(15) Dissolving the colquhounod D derivative 30 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol and water;
(16) Under the inert gas atmosphere, adding an azide reagent, sodium ascorbate and copper sulfate pentahydrate, stirring at room temperature to react until TLC (thin layer chromatography) shows that the reaction is complete, and diluting the reaction liquid with an organic solvent, extracting, drying, concentrating in vacuum and purifying to obtain the colquhounod D derivative 35.
5. A process for the preparation of the colquhounod D derivative 39-47, which is a colquhounod, as claimed in claim 2, characterized in that it comprises the following steps:
(1) Dissolving a compound colquhounod D in an organic solvent N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol; adding 2M hydrochloric acid in inert gas atmosphere, stirring at 45 ℃ for reaction until TLC (thin layer chromatography) shows that the reaction is complete, and obtaining a colquhouoid D derivative 39-40 after diluting, washing, drying, filtering, concentrating in vacuum and purifying the mixture;
(2) Dissolving a compound colquhounod D in an organic solvent N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol; adding sodium hydride, halogenated hydrocarbon methyl iodide, bromopropene, bromopropyne and benzyl bromide in inert gas atmosphere, heating to 25-60 ℃, stirring to react until TLC shows complete reaction, filtering the reaction liquid, collecting filtrate, concentrating in vacuum, and purifying to obtain a colquhouoid D derivative 41-44;
(3) Dissolving the colquhounod D derivative 43 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol; adding 1.5M diisobutyl aluminum hydride in inert gas atmosphere, stirring at 0-60 ℃ to react until TLC shows complete reaction, quenching the reaction liquid with potassium sodium tartrate, extracting, drying, concentrating in vacuum, purifying to obtain an intermediate;
(4) Dissolving the intermediate in an organic solvent N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine or ethanol; adding N, N-thiocarbonyldiimidazole and DMAP under inert gas atmosphere, heating to 60-80 ℃, stirring until TLC shows that the reaction is complete, and vacuum concentrating and purifying the reaction liquid to obtain a colquhounod D derivative 45-46;
(5) Dissolving the colquhounod D derivative 45 in an organic solvent of N, N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, acetonitrile, toluene, pyridine, ethanol and water; under the inert gas atmosphere, adding an azide reagent, sodium ascorbate and copper sulfate pentahydrate, stirring at room temperature to react until TLC (thin layer chromatography) shows that the reaction is complete, and diluting the reaction liquid with an organic solvent, extracting, drying, concentrating in vacuum and purifying to obtain the colquhounod D derivative 47.
6. Use of a labdane sesterterpene colquhounod D derivative according to claim 1 or 2 for the preparation of an anti-inflammatory or immunosuppressive drug.
7. Use of a labdane sesterterpene colquhounod D derivative according to claim 1 or 2 for the preparation of a medicament for the treatment of ulcerative colitis.
8. A pharmaceutical composition comprising a pharmaceutically acceptable pharmaceutical adjuvant and any one or any combination of the labdane sesterterpene colquhounod D derivatives of claims 1 or 2.
9. The pharmaceutical composition according to claim 8, wherein the pharmaceutical composition is in the form of granules, powder spray, patch, tablet, capsule, granule, oral liquid, injection.
10. The use of the pharmaceutical composition of claim 8 in the manufacture of an anti-inflammatory or immunosuppressive drug, or in the manufacture of a medicament for the treatment of ulcerative colitis.
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