CN115746079B - 一种薯蓣皂苷元衍生物及其制备方法与用途 - Google Patents
一种薯蓣皂苷元衍生物及其制备方法与用途 Download PDFInfo
- Publication number
- CN115746079B CN115746079B CN202211393095.9A CN202211393095A CN115746079B CN 115746079 B CN115746079 B CN 115746079B CN 202211393095 A CN202211393095 A CN 202211393095A CN 115746079 B CN115746079 B CN 115746079B
- Authority
- CN
- China
- Prior art keywords
- compound
- derivative
- nmr
- diosgenin
- cdcl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- WQLVFSAGQJTQCK-VKROHFNGSA-N diosgenin Chemical class O([C@@H]1[C@@H]([C@]2(CC[C@@H]3[C@@]4(C)CC[C@H](O)CC4=CC[C@H]3[C@@H]2C1)C)[C@@H]1C)[C@]11CC[C@@H](C)CO1 WQLVFSAGQJTQCK-VKROHFNGSA-N 0.000 title claims abstract description 67
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- DWCSNWXARWMZTG-UHFFFAOYSA-N Trigonegenin A Natural products CC1C(C2(CCC3C4(C)CCC(O)C=C4CCC3C2C2)C)C2OC11CCC(C)CO1 DWCSNWXARWMZTG-UHFFFAOYSA-N 0.000 claims abstract description 38
- WQLVFSAGQJTQCK-UHFFFAOYSA-N diosgenin Natural products CC1C(C2(CCC3C4(C)CCC(O)CC4=CCC3C2C2)C)C2OC11CCC(C)CO1 WQLVFSAGQJTQCK-UHFFFAOYSA-N 0.000 claims abstract description 38
- 239000003814 drug Substances 0.000 claims abstract description 21
- 150000001875 compounds Chemical class 0.000 claims description 35
- 125000000217 alkyl group Chemical group 0.000 claims description 16
- 229910052736 halogen Inorganic materials 0.000 claims description 14
- 150000002367 halogens Chemical group 0.000 claims description 14
- 229910052739 hydrogen Inorganic materials 0.000 claims description 14
- 239000001257 hydrogen Substances 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 9
- 150000003839 salts Chemical class 0.000 claims description 9
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 8
- 238000000855 fermentation Methods 0.000 claims description 8
- 230000004151 fermentation Effects 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 7
- 230000004770 neurodegeneration Effects 0.000 claims description 7
- 208000015122 neurodegenerative disease Diseases 0.000 claims description 7
- 150000002431 hydrogen Chemical class 0.000 claims description 6
- 230000000324 neuroprotective effect Effects 0.000 claims description 6
- 208000024827 Alzheimer disease Diseases 0.000 claims description 4
- 208000018737 Parkinson disease Diseases 0.000 claims description 4
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 4
- 239000000758 substrate Substances 0.000 claims description 4
- 241000194107 Bacillus megaterium Species 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 125000002023 trifluoromethyl group Chemical group FC(F)(F)* 0.000 claims 1
- 230000004112 neuroprotection Effects 0.000 abstract description 10
- 230000001681 protective effect Effects 0.000 abstract description 9
- 229940079593 drug Drugs 0.000 abstract description 7
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 238000001308 synthesis method Methods 0.000 abstract description 3
- 238000005481 NMR spectroscopy Methods 0.000 description 71
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 30
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 27
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 23
- 210000004027 cell Anatomy 0.000 description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 17
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 16
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 12
- 230000015572 biosynthetic process Effects 0.000 description 10
- 230000004224 protection Effects 0.000 description 10
- 238000003786 synthesis reaction Methods 0.000 description 10
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 8
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 8
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 8
- TWZIEWLIAZSJHV-UHFFFAOYSA-N 7β-hydroxydiosgenin Chemical compound CC1C(C2(CCC3C4(C)CCC(O)CC4=CC(O)C3C2C2)C)C2OC11CCC(C)CO1 TWZIEWLIAZSJHV-UHFFFAOYSA-N 0.000 description 8
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 8
- 239000012074 organic phase Substances 0.000 description 7
- 238000003756 stirring Methods 0.000 description 7
- 239000001963 growth medium Substances 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 239000003208 petroleum Substances 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- 239000000376 reactant Substances 0.000 description 6
- 238000000926 separation method Methods 0.000 description 6
- 238000004809 thin layer chromatography Methods 0.000 description 6
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 5
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 5
- 238000010898 silica gel chromatography Methods 0.000 description 5
- 125000003545 alkoxy group Chemical group 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 239000012044 organic layer Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 101100223822 Dictyostelium discoideum zfaA gene Proteins 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 239000002552 dosage form Substances 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- -1 steroid compound Chemical class 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 108010087230 Sincalide Proteins 0.000 description 2
- 230000003376 axonal effect Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 238000010609 cell counting kit-8 assay Methods 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 230000007850 degeneration Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 125000002816 methylsulfanyl group Chemical group [H]C([H])([H])S[*] 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- ANRMAUMHJREENI-ZZXKWVIFSA-N (E)-4-(trifluoromethyl)cinnamic acid Chemical compound OC(=O)\C=C\C1=CC=C(C(F)(F)F)C=C1 ANRMAUMHJREENI-ZZXKWVIFSA-N 0.000 description 1
- XNCRUNXWPDJHGV-BQYQJAHWSA-N (e)-2-methyl-3-phenylprop-2-enoic acid Chemical compound OC(=O)C(/C)=C/C1=CC=CC=C1 XNCRUNXWPDJHGV-BQYQJAHWSA-N 0.000 description 1
- CPDDDTNAMBSPRN-ZZXKWVIFSA-N (e)-3-(4-bromophenyl)prop-2-enoic acid Chemical compound OC(=O)\C=C\C1=CC=C(Br)C=C1 CPDDDTNAMBSPRN-ZZXKWVIFSA-N 0.000 description 1
- LBOUHDMYVURTMA-AATRIKPKSA-N (e)-3-(4-formylphenyl)prop-2-enoic acid Chemical compound OC(=O)\C=C\C1=CC=C(C=O)C=C1 LBOUHDMYVURTMA-AATRIKPKSA-N 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- BBQDLDVSEDAYAA-AATRIKPKSA-N 2-nitrocinnamic acid Chemical compound OC(=O)\C=C\C1=CC=CC=C1[N+]([O-])=O BBQDLDVSEDAYAA-AATRIKPKSA-N 0.000 description 1
- SCJHPUGWYHCYAZ-UHFFFAOYSA-N 3,5-diethyl-2-propylpyridine Chemical compound CCCC1=NC=C(CC)C=C1CC SCJHPUGWYHCYAZ-UHFFFAOYSA-N 0.000 description 1
- ISMMYAZSUSYVQG-ZZXKWVIFSA-N 4-Fluorocinnamic acid Chemical compound OC(=O)\C=C\C1=CC=C(F)C=C1 ISMMYAZSUSYVQG-ZZXKWVIFSA-N 0.000 description 1
- RURHILYUWQEGOS-VOTSOKGWSA-N 4-Methylcinnamic acid Chemical compound CC1=CC=C(\C=C\C(O)=O)C=C1 RURHILYUWQEGOS-VOTSOKGWSA-N 0.000 description 1
- GXLIFJYFGMHYDY-ZZXKWVIFSA-N 4-chlorocinnamic acid Chemical compound OC(=O)\C=C\C1=CC=C(Cl)C=C1 GXLIFJYFGMHYDY-ZZXKWVIFSA-N 0.000 description 1
- 206010003694 Atrophy Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 description 1
- 241000234272 Dioscoreaceae Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 208000026139 Memory disease Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010029260 Neuroblastoma Diseases 0.000 description 1
- 206010067482 No adverse event Diseases 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000037444 atrophy Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 208000015114 central nervous system disease Diseases 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 229930016911 cinnamic acid Natural products 0.000 description 1
- 235000013985 cinnamic acid Nutrition 0.000 description 1
- 230000003920 cognitive function Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- ROAYSRAUMPWBQX-UHFFFAOYSA-N ethanol;sulfuric acid Chemical compound CCO.OS(O)(=O)=O ROAYSRAUMPWBQX-UHFFFAOYSA-N 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000005802 health problem Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000008297 liquid dosage form Substances 0.000 description 1
- 206010027175 memory impairment Diseases 0.000 description 1
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000003007 myelin sheath Anatomy 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 239000007909 solid dosage form Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
本发明提供了一种薯蓣皂苷元衍生物及其制备方法与用途,属于生物医药技术领域。该薯蓣皂苷元衍生物的结构如式I所示。本发明提供的薯蓣皂苷元衍生物对于过氧化氢损伤的SH‑SY5Y细胞具有优良的保护作用,说明该薯蓣皂苷元衍生物具有优良的神经保护作用,并且,其中DG2、DG3、DG4、DG6、DG7、DG10‑DG18的保护作用优于薯蓣皂苷元。本发明提供的薯蓣皂苷元衍生物的结构新颖,合成方法简单高效,成本低,在制备具有神经保护作用的药物中具有广阔的应用前景。
Description
技术领域
本发明属于生物医药技术领域,具体涉及一种薯蓣皂苷元衍生物及其制备方法与用途。
背景技术
随着人口老龄化的加剧,神经退行性疾病的发病率也在不断攀升。神经退行性疾病(Neurodegenerative disease)是指由于神经元或其髓鞘的丧失导致人体出现功能障碍的中枢神经系统疾病,其特征是中枢神经系统细胞的进行性退化和死亡,其中阿尔茨海默病(AD)、帕金森病(PD)是患病率最高的两种慢性神经退行性疾病,给人类健康问题带来了极大痛苦,开发高效的神经保护候选药物迫在眉睫。
薯蓣皂苷元是一种甾体类化合物,主要存在于薯蓣科等植物中,是甾体类激素药物的重要生产前体,被证明具有抗肿瘤、抗氧化及神经保护等广泛的药理学作用。研究表明,薯蓣皂苷元可通过预防Aβ诱导的轴突萎缩和记忆缺陷来改善认知功能并显示出有效的神经保护作用(Diosgenin restores Aβ-induced axonal degeneration by reducingthe expression of heat shock cognate 70(HSC70).Scientific reports,2018)。但是,一方面,由于薯蓣皂苷元本身的水溶性差、疏水性强及生物利用度低等不足限制了其在临床上的应用;另一方面,薯蓣皂苷元的神经保护作用还有待进一步提高。
为了克服上述问题,亟需开发出神经保护作用更优异的、适合临床应用的药物。
发明内容
本发明的目的在于提供一种新的薯蓣皂苷元衍生物及其制备方法与用途。
本发明提供了式I所示化合物、其盐或其立体异构体:
其中,R1为氢或R2;
R2为
m选自0-5的整数;
R3各自独立地选自氢、C1-5烷基、卤素取代的C1-5烷基、C1-5烷氧基、卤素取代的C1-5烷氧基、硝基、卤素、-CHO、甲硫基;
R4为氢或C1-5烷基;
R5为氢或C1-5烷基。
进一步地,所述化合物的结构如式II所示:
R1、R2如上所述。
进一步地,所述化合物的结构如式III或式IV所示:
其中,R2为
m选自0、1、2或3;
R3各自独立地选自氢、C1-3烷基、卤素取代的C1-3烷基、C1-3烷氧基、卤素取代的C1-3烷氧基、硝基、卤素、-CHO、甲硫基;
R4为氢或C1-3烷基。
进一步地,R2为
所述卤素为F、Cl或Br;
所述卤素取代的C1-3烷基为三氟甲基。
进一步地,R2选自以下基团:
进一步地,所述化合物选自:
本发明还提供了一种制备上述化合物的方法,所述方法包括以下步骤:
式I-a所示化合物与式I-b所示化合物反应,得到式I所示化合物;
其中,R1、R2、R3、R4、R5、m如上所述;
优选地,所述式I-a所示化合物的制备方法包括以下步骤:薯蓣皂苷元为底物与巨大芽孢杆菌共培养,发酵结束后收集发酵上清,提纯,得到式I-a所示化合物。
本发明还提供了一种具有神经保护作用的药物,它是以上述化合物、其盐或其立体异构体为活性成分,加上药学上可接受的辅料制备得到的制剂。
本发明还提供了上述化合物、其盐或其立体异构体在制备具有神经保护作用的药物中的用途。
进一步地,所述药物是预防和/或治疗神经退行性疾病的药物,所述神经退行性疾病优选为阿尔兹海默症、帕金森。
本发明提供的药物可以为多种剂型,并以单位剂量形式给药。所述药物剂型或给药剂型可以是液体剂型、固体剂型、外用制剂、喷剂等。
本发明提供的药物中还可以含有常用的载体,这里所述可药用载体包括但不局限于:微囊与微球、纳米粒、脂质体。
与现有技术相比,本发明取得了以下有益效果:
1.本发明提供的薯蓣皂苷元衍生物对于过氧化氢损伤的SH-SY5Y细胞具有优良的保护作用,说明该薯蓣皂苷元衍生物具有优良的神经保护作用,并且,其中DG2、DG3、DG4、DG6、DG7、DG10-DG18的保护作用优于薯蓣皂苷元;
2.本发明提供的薯蓣皂苷元衍生物的结构新颖,此前未见报道;
3.本发明提供的薯蓣皂苷元衍生物的合成方法简单高效,成本低。
综上,本发明提供的薯蓣皂苷元衍生物在制备具有神经保护作用的药物中具有广阔的应用前景。
显然,根据本发明的上述内容,按照本领域的普通技术知识和惯用手段,在不脱离本发明上述基本技术思想前提下,还可以做出其它多种形式的修改、替换或变更。
以下通过实施例形式的具体实施方式,对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
附图说明
图1:衍生物DG1的1H NMR(600MHz,CDCl3)图。
图2:衍生物DG1的13C NMR(150MHz,CDCl3)图。
图3:衍生物DG1的HR-ESI-MS图。
图4:衍生物DG2的1H NMR(600MHz,CDCl3)图。
图5:衍生物DG2的13C NMR(150MHz,CDCl3)图。
图6:衍生物DG2的HR-ESI-MS图。
图7:衍生物DG3的1H NMR(600MHz,CDCl3)图。
图8:衍生物DG3的13C NMR(150MHz,CDCl3)图。
图9:衍生物DG3的HR-ESI-MS图。
图10:衍生物DG4的1H NMR(600MHz,CDCl3)图。
图11:衍生物DG4的13C NMR(150MHz,CDCl3)图。
图12:衍生物DG4的HR-ESI-MS图。
图13:衍生物DG5的1H NMR(600MHz,CDCl3)图。
图14:衍生物DG5的13C NMR(150MHz,CDCl3)图。
图15:衍生物DG5的HR-ESI-MS图。
图16:衍生物DG6的1H NMR(600MHz,CDCl3)图。
图17:衍生物DG6的13C NMR(150MHz,CDCl3)图。
图18:衍生物DG6的HR-ESI-MS图。
图19:衍生物DG7的1H NMR(600MHz,CDCl3)图。
图20:衍生物DG7的13C NMR(150MHz,CDCl3)图。
图21:衍生物DG7的HR-ESI-MS图。
图22:衍生物DG8的1H NMR(600MHz,CDCl3)图。
图23:衍生物DG8的13C NMR(150MHz,CDCl3)图。
图24:衍生物DG8的HR-ESI-MS图。
图25:衍生物DG9的1H NMR(600MHz,CDCl3)图。
图26:衍生物DG9的13C NMR(150MHz,CDCl3)图。
图27:衍生物DG9的HR-ESI-MS图。
图28:衍生物DG10的1H NMR(600MHz,CDCl3)图。
图29:衍生物DG10的13C NMR(150MHz,CDCl3)图。
图30:衍生物DG10的HR-ESI-MS图。
图31:衍生物DG11的1H NMR(600MHz,CDCl3)图。
图32:衍生物DG11的13C NMR(150MHz,CDCl3)图。
图33:衍生物DG11的HR-ESI-MS图。
图34:衍生物DG12的1H NMR(600MHz,CDCl3)图。
图35:衍生物DG12的13C NMR(150MHz,CDCl3)图。
图36:衍生物DG12的HR-ESI-MS图。
图37:衍生物DG13的1H NMR(600MHz,CDCl3)图。
图38:衍生物DG13的13C NMR(150MHz,CDCl3)图。
图39:衍生物DG13的HR-ESI-MS图。
图40:衍生物DG14的1H NMR(600MHz,CDCl3)图。
图41:衍生物DG14的13C NMR(150MHz,CDCl3)图。
图42:衍生物DG14的HR-ESI-MS图。
图43:衍生物DG15的1H NMR(600MHz,CDCl3)图。
图44:衍生物DG15的13C NMR(150MHz,CDCl3)图。
图45:衍生物DG15的HR-ESI-MS图。
图46:衍生物DG16的1H NMR(600MHz,CDCl3)图。
图47:衍生物DG16的13C NMR(150MHz,CDCl3)图。
图48:衍生物DG16的HR-ESI-MS图。
图49:衍生物DG17的1H NMR(600MHz,CDCl3)图。
图50:衍生物DG17的13C NMR(150MHz,CDCl3)图。
图51:衍生物DG17的HR-ESI-MS图。
图52:衍生物DG18的1H NMR(600MHz,CDCl3)图。
图53:衍生物DG18的13C NMR(150MHz,CDCl3)图。
图54:衍生物DG18的HR-ESI-MS图。
图55:薯蓣皂苷元以及衍生物对SH-SY5Y细胞的细胞毒性作用。
具体实施方式
如无特别说明,本发明所用原料与设备均为已知产品,通过购买市售产品所得。
7β-羟基薯蓣皂苷元可以按照本发明实施例1提供的方法制备得到,也可以通过本领域技术人员公知的其它方法合成得到。
实施例1、制备薯蓣皂苷元衍生物DG1-DG18
步骤一:制备7β-羟基薯蓣皂苷元
取500mg薯蓣皂苷元,溶于50mL DMSO,配置成10mg/mL的底物溶液。采用二步培养法,先把巨大芽孢杆菌B.megateriumCGMCC 1.1741由斜面接入无菌的豆粕液体培养基(培养基配方:市售豆粕5g,葡萄糖20g,酵母浸膏5g,NaCl 5g,K2HPO4 5g,蒸馏水定容至1L)中培养巨大芽孢杆菌至对数生长期(培养温度28℃,摇床转速180rpm·min-1),按0.8mL/瓶的接种量转接至新鲜的无菌液体培养基中,继续培养24h后每瓶添加10mg上述底物溶液(控制薯蓣皂苷元的终浓度为10mg/250mL),继续培养96h后结束发酵,取发酵上清以等体积的乙酸乙酯萃取3遍,减压回收有机溶剂,得到薯蓣皂苷元中间体发酵产物。
进一步地,将上述的薯蓣皂苷元中间体发酵产物进行分离纯化,萃取液经旋蒸浓缩后用甲醇溶解,加入样品量约2倍量的硅胶(100-200目)拌样,置于60℃水浴锅上挥干有机相。再用30倍量的硅胶(200-300目)进行柱层析,以石油醚:乙酸乙酯体系极性梯度洗脱(石油醚︰乙酸乙酯的体积比为10︰1→1︰1),TLC分析,10%硫酸乙醇加热显色,合并相似馏分,极性相近而难以用硅胶柱层析分离纯化的组分利用制备液相进一步纯化,得到7β-羟基薯蓣皂苷元。
步骤二:制备薯蓣皂苷元衍生物DG1-DG18
试剂和条件:(a)DG1-DG2、DG16-DG20:CH2Cl2,DMAP,EDCI,60℃,12h;(b)DG3-DG4:CH2Cl2,DMAP,EDCI,rt,12h;(c)DG5-DG15、DG21-DG22:CH2Cl2,DMAP,EDCI,60℃,18h。
按照上述路线,制备得到薯蓣皂苷元衍生物DG1-DG18。薯蓣皂苷元衍生物DG1-DG18的结构如表1所示。
表1.薯蓣皂苷元衍生物DG1-DG18的结构
/>
制备薯蓣皂苷元衍生物DG1-18的具体操作如下:
(1)制备DG1-DG2:
称取200mg(0.483mmol,1equiv)7β-羟基薯蓣皂苷元置于25ml茄形瓶中,在依次加入214.45mg(1.449mmol,3equiv)肉桂酸,117.85mg(0.966mmol,2equiv)DMAP,184.5mg(0.966mmol,2equiv)EDCI后,加入12ml无水二氯甲烷进行充分溶解,于60℃中加热搅拌反应12h,薄层色谱法检测合成情况,合成结束后,取反应物在冰浴下缓慢滴加适量的4℃预冷的饱和碳酸氢钠,滴加完成后撤掉冰浴在常温下搅拌30min,用水和二氯甲烷萃取反应物,连续萃取3遍,合并下层有机相,将有机层与无水硫酸钠混合干燥,过滤,减压回收溶剂后,通过硅胶柱层析(石油醚∶乙酸乙酯)进行分离纯化,分别得到白色化合物60mg/70mg,产率为30%/35%。
DG1:HR-ESI-MSm/z:691.3864[M+H]+(分子式经计算为C45H55O6,691.3999)。1H NMR(600MHz,CDCl3)δ7.68(dd,J=16.0,8.1Hz,2H),7.59–7.46(m,4H),7.46–7.32(m,6H),6.42(dd,J=16.0,4.4Hz,2H),5.37(t,J=3.6Hz,1H),5.18(dd,J=8.7,2.1Hz,1H),4.77(dd,J=10.7,6.0,4.4Hz,1H),4.37(ddd,1H),3.50–3.39(m,1H),3.32(t,J=11.0Hz,1H),2.53–2.34(m,2H),0.97(d,J=6.9Hz,3H),0.84(s,3H),0.75(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ166.97,166.23,144.90,144.67,144.31,134.48,134.42,130.29,130.24,128.88(C×4),128.18(C×2),128.06(C×2),122.26,118.49,118.46,109.24,80.92,75.50,73.27,66.81,61.44,55.15,48.00,41.66,40.74,39.44,37.68,36.77,36.60,36.44,33.27,31.43(d,J=27.3Hz),30.22(d,J=19.8Hz),28.79,27.76,20.90,19.11,17.09,16.25,14.55.
DG2:HR-ESI-MSm/z:561.3548[M+H]+(分子式经计算为C36H49O5,561.3508)。1H NMR(600MHz,CDCl3)δ7.67(d,J=16.0Hz,1H),7.56–7.48(m,2H),7.38(dd,J=4.9,1.9Hz,3H),6.42(d,J=16.0Hz,1H),5.35(t,J=2.0Hz,1H),4.87–4.68(m,1H),4.45(ddd,J=8.7,7.5,6.4Hz,1H),3.91–3.74(m,1H),3.53–3.43(m,1H),3.39(t,J=11.0Hz,1H),2.53–2.35(m,2H),2.35–2.18(m,1H),1.11(s,3H),0.98(d,J=6.9Hz,3H),0.81(s,3H),0.79(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ166.36,144.68,142.38,134.46,130.26,128.89(C×2),128.07(C×2),126.41,118.48,109.28,81.06,73.50,73.00,66.89,61.56,55.46,48.01,41.68,40.83,40.69,39.58,37.73,36.74,36.69,34.28,31.41,30.32,28.81,27.80,20.87,19.17,17.16,16.27,14.58.
(2)制备DG3-DG4:
称取500mg(1.208mmol,1equiv)7β-羟基薯蓣皂苷元置于50ml茄形瓶中,在依次加入330.87mg(1.812mmol,1.5equiv)4-氯肉桂酸,147.38mg(1.208mmol,1equiv)DMAP,346.09mg(1.812mmol,1.5equiv)EDCI后,加入20ml无水二氯甲烷进行充分溶解,常温反应,薄层色谱法检测合成情况,合成结束后,取反应物在冰浴下缓慢滴加适量的4℃预冷的饱和碳酸氢钠,滴加完成后撤掉冰浴在常温下搅拌30min,用水和二氯甲烷萃取反应物,连续萃取3遍,合并下层有机相,将有机层与无水硫酸钠混合干燥,过滤,减压回收溶剂后,通过硅胶柱层析(石油醚∶乙酸乙酯)进行分离纯化,分别得到白色化合物150mg/175mg,产率为30%/35%。
DG3:HR-ESI-MSm/z:759.3271[M+H]+(分子式经计算为C45H53Cl2O6,759.3219)。1HNMR(600MHz,CDCl3)δ7.61(dd,J=16.0,6.0Hz,2H),7.48–7.42(m,4H),7.36(t,J=8.3Hz,4H),6.38(dd,J=16.0,1.8Hz,2H),5.35(d,J=1.8Hz,1H),5.23–5.11(m,1H),4.81–4.66(m,1H),4.37(ddd,J=8.7,7.6,6.4Hz,1H),3.54–3.38(m,1H),3.31(t,J=11.0Hz,1H),2.51–2.36(m,2H),1.16(s,3H),0.97(d,J=6.9Hz,3H),0.84(s,3H),0.75(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ166.71,165.97,144.36,143.43,143.22,136.20,136.15,132.95,132.90,129.32(C×4),129.21(C×2),129.18(C×2),122.17,119.06,119.03,109.28,80.91,75.61,73.38,66.84,61.41,55.14,48.00,41.66,40.73,39.42,37.66,36.76,36.57,36.41,33.26,31.32,30.27,29.72,28.80,27.73,20.89,19.09,17.09,16.24,14.53.
DG4:HR-ESI-MSm/z:595.3144[M+H]+(分子式经计算为C36H48ClO5,595.3190)。1HNMR(600MHz,CDCl3)δ7.61(d,J=16.0Hz,1H),7.50–7.42(m,2H),7.40–7.31(m,2H),6.38(d,J=16.0Hz,1H),5.34(t,J=2.0Hz,1H),4.85–4.65(m,1H),4.45(ddd,J=8.7,7.5,6.4Hz,1H),3.89–3.77(m,1H),3.48(ddd,J=10.9,4.4,2.1Hz,1H),3.39(t,J=11.0Hz,1H),2.50–2.38(m,2H),2.36–2.14(m,1H),1.11(s,3H),0.98(d,J=6.9Hz,3H),0.81(s,3H),0.79(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ166.09,143.21,142.30,136.16,132.95,129.22(C×2),129.19(C×2),126.45,119.07,109.28,81.05,73.64,72.98,66.89,61.56,55.45,48.00,41.68,40.83,40.68,39.57,37.71,36.74,36.67,34.28,31.42,30.32,28.81,27.79,20.86,19.17,17.16,16.26,14.57.
(3)制备DG5-12:
称取200mg(0.483mmol,1equiv)7β-羟基薯蓣皂苷元置于50ml茄形瓶中,在依次加入117.85mg(2.426mmol,2equiv)DMAP,184.51mg(0.966mmol,2equiv)EDCI,328.92mg(1.449mmol,3equiv)4-溴肉桂酸/312.98mg(1.449mmol,3equiv)4-(三氟甲基)肉桂酸/234.74mg(1.449mmol,3equiv)4-甲基肉桂酸/279.66mg(1.449mmol,3equiv)4-硝基肉桂酸后,加入20ml无水二氯甲烷进行充分溶解,60℃加热搅拌18h,薄层色谱法检测合成情况,合成结束后,取反应物在冰浴下缓慢滴加适量的4℃预冷的饱和碳酸氢钠,滴加完成后撤掉冰浴在常温下搅拌30min,用水和二氯甲烷萃取反应物,连续萃取3遍,合并下层有机相,将有机层与无水硫酸钠混合干燥,过滤,减压回收溶剂后,通过硅胶柱层析(石油醚∶乙酸乙酯)进行分离纯化,产率为60%/35%/60%/35%/50%/40%/45%/45%。
DG5:HR-ESI-MSm/z:847.2226[M+H]+(分子式经计算为C45H53Br2O6,847.2209)。1HNMR(600MHz,CDCl3)δ7.60(dd,J=16.0,6.1Hz,2H),7.51(t,J=8.2Hz,4H),7.44–7.30(m,4H),6.39(dd,J=16.0,1.8Hz,2H),5.35(t,J=1.8Hz,1H),5.17(dt,J=8.7,2.1Hz,1H),4.81–4.65(m,1H),4.37(ddd,J=8.8,7.6,6.4Hz,1H),3.44(ddd,J=10.9,4.4,2.1Hz,1H),3.31(t,J=11.0Hz,1H),2.55–2.33(m,2H),1.16(s,3H),0.97(d,J=6.9Hz,3H),0.83(s,3H),0.75(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ166.69,165.96,144.37,143.50,143.29,133.37,133.32,132.14(C×4),129.54(C×2),129.43(C×2),124.53(d,J=9.7Hz),122.16,119.17,119.14,109.28,80.91,75.63,73.40,66.84,61.41,55.13,47.99,41.66,40.73,39.42,37.65,36.76,36.49(d,J=24.1Hz),33.26,31.52,31.32,30.21(d,J=18.2Hz),29.72,28.80,27.72,20.89,19.09,17.09,16.24,14.53.
DG6:HR-ESI-MSm/z:639.2526[M+H]+(分子式经计算为C36H48BrO5,639.2685)。1HNMR(600MHz,CDCl3)δ7.60(d,J=16.1Hz,1H),7.56–7.48(m,2H),7.44–7.34(m,2H),6.40(d,J=16.0Hz,1H),5.34(t,J=2.0Hz,1H),4.84–4.65(m,1H),4.45(ddd,J=8.9,7.5,6.3Hz,1H),3.85(dt,J=8.2,2.3Hz,1H),3.48(ddd,J=11.0,4.5,2.1Hz,1H),3.39(t,J=11.0Hz,1H),2.51–2.34(m,2H),1.11(s,3H),0.98(d,J=6.9Hz,3H),0.81(s,3H),0.79(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ166.07,143.27,142.30,133.38,132.15(C×2),129.44(C×2),126.45,124.50,119.19,109.28,81.05,73.66,72.98,66.89,61.56,55.45,48.00,41.68,40.83,40.68,39.57,37.70,36.74,36.67,34.28,31.42,30.32,28.81,27.78,20.86,19.17,17.16,16.26,14.58.
DG7:HR-ESI-MSm/z:827.1719[M+H]+(分子式经计算为C47H53F6O6,827.3746)。1HNMR(600MHz,CDCl3)δ7.69(q,J=1.5Hz,1H),7.42(d,J=3.3Hz,3H),7.29(s,0H),5.43(dd,J=4.6,2.6Hz,1H),4.85–4.63(m,1H),4.44(ddd,J=8.7,7.5,6.4Hz,1H),3.50(ddd,J=10.9,4.4,2.1Hz,1H),3.40(t,J=11.0Hz,1H),2.50–2.40(m,2H),2.13(d,J=1.5Hz,3H),1.10(s,3H),1.00(d,J=6.9Hz,3H),0.82(d,J=1.3Hz,4H),0.81(s,2H).13C NMR(150MHz,CDCl3)δ166.40,165.65,144.46,143.02,142.80,137.82,137.76,131.88,131.67,128.27(C×4),128.16(C×4),125.91,125.85,122.09,121.04,121.01,109.29,80.89,75.83,73.58,66.85,61.41,55.14,48.00,41.67,40.74,39.41,37.64,36.77,36.56,36.41,33.27,31.42(d,J=29.7Hz),30.20(d,J=17.3Hz),28.80,27.70,20.90,19.08,17.09,16.24,14.53.
DG8:HR-ESI-MSm/z:629.3413[M+H]+(分子式经计算为C37H48F3O5,629.3454)。1HNMR(600MHz,CDCl3)δ7.70(d,J=16.0Hz,1H),7.68–7.62(m,4H),6.51(d,J=16.0Hz,1H),5.38(t,J=2.0Hz,1H),4.89–4.66(m,1H),4.48(ddd,J=8.8,7.5,6.4Hz,1H),3.88(dt,J=8.1,2.2Hz,1H),3.50(ddd,J=10.9,4.4,2.1Hz,1H),3.41(t,J=11.0Hz,1H),2.51–2.39(m,2H),2.34–2.25(m,1H),1.39(s,1H),1.31(s,2H),1.14(s,3H),1.01(d,J=7.0Hz,3H),0.84(s,3H),0.82(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ165.76,142.77,142.23,137.83,131.64,128.17(C×4),126.52,125.88(d,J=3.4Hz),121.07,109.29,81.05,73.86,72.98,66.89,61.57,55.45,48.00,41.68,40.83,40.68,39.57,37.68,36.74,36.66,34.28,31.47(d,J=15.2Hz),30.23(d,J=26.0Hz),28.81,27.77,20.87,19.17,17.16,16.26,14.57.
DG9:HR-ESI-MS m/z:719.4874[M+H]+(分子式经计算为C47H59O6,719.4312)。1HNMR(600MHz,CDCl3)δ7.65(dd,J=15.9,7.6Hz,2H),7.42(dd,J=8.2,6.6Hz,4H),7.19(dd,J=8.0,6.7Hz,4H),6.37(dd,J=16.0,3.9Hz,2H),5.36(t,J=1.8Hz,1H),5.18(dt,J=8.8,2.1Hz,1H),4.76(tdd,J=10.7,6.1,4.4Hz,1H),4.37(ddd,J=8.8,7.5,6.4Hz,1H),3.44(ddd,J=10.9,4.4,2.1Hz,1H),3.32(t,J=11.0Hz,1H),2.37(d,J=5.9Hz,6H),1.16(s,3H),0.97(d,J=7.0Hz,3H),0.84(s,3H),0.75(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ167.17,166.43,144.90,144.66,144.26,140.69,140.64,131.75,131.69,129.62(C×4),128.17(C×2),128.06(C×2),122.30,117.37,117.36,109.24,80.93,75.38,73.17,66.81,61.44,55.15,48.00,41.66,40.73,39.44,37.69,36.77,36.61,36.44,33.27,31.43(d,J=27.3Hz),30.31(d,J=7.4Hz),29.72,28.79,27.77,21.51,21.48,20.90,19.10,17.09,16.25,14.54.
DG10:HR-ESI-MSm/z:575.3635[M+H]+(分子式经计算为C37H51O5,575.3736)。1HNMR(600MHz,CDCl3)δ7.65(d,J=16.0Hz,1H),7.47–7.38(m,2H),7.19(d,J=7.9Hz,2H),6.37(d,J=16.0Hz,1H),5.34(t,J=2.0Hz,1H),4.89–4.58(m,1H),4.45(ddd,J=8.7,7.5,6.3Hz,1H),3.85(dt,J=8.2,2.3Hz,1H),3.60–3.39(m,1H),2.37(s,3H),1.11(s,3H),0.98(d,J=7.0Hz,3H),0.81(s,3H),0.79(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ166.56,144.68,142.42,140.68,131.73,129.63(C×2),128.07(C×2),126.37,117.36,109.28,81.06,73.39,73.00,66.88,61.56,55.46,48.01,41.68,40.83,40.69,39.58,37.75,36.74,36.70,34.28,31.47(d,J=16.2Hz),30.23(d,J=26.1Hz).,28.81,27.82,21.49,20.86,19.17,17.16,16.27,14.58.
DG11:HR-ESI-MSm/z:781.3673[M+H]+(分子式经计算为C45H53N2O10,781.3700)。1HNMR(600MHz,CDCl3)δ8.32–8.20(m,4H),7.71(d,J=4.3Hz,1H),7.69–7.59(m,6H),6.53(dd,J=16.0,1.1Hz,2H),5.36(t,J=1.9Hz,1H),5.20(dd,J=8.7,2.1Hz,1H),4.78(tdd,J=10.6,5.8,4.3Hz,1H),4.38(ddd,J=8.6,7.5,6.3Hz,1H),3.45(ddd,J=10.9,4.5,2.2Hz,1H),3.30(t,J=11.0Hz,1H),2.48–2.33(m,2H),1.17(s,3H),1.09(d,J=17.8Hz,1H),0.97(d,J=6.9Hz,4H),0.84(s,3H),0.75(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ166.01,165.27,148.53,148.50,144.56,141.92,141.74,140.55,140.50,128.73(C×2),128.62(C×2),124.19(C×4),122.77,122.73,121.99,109.33,80.88,76.05,73.78,66.87,61.39,55.12,47.99,41.68,40.74,39.39,37.62,36.77,36.53,36.39,33.27,31.30,30.24,28.81,27.68,20.90,19.07,17.09,16.24,14.52.
DG12:HR-ESI-MSm/z:606.3427[M+H]+(分子式经计算为C36H48NO7,606.3431)。1HNMR(600MHz,CDCl3)δ8.25(d,J=8.7Hz,1H),7.74–7.62(m,3H),6.54(d,J=16.0Hz,1H),5.35(t,J=1.9Hz,1H),4.87–4.70(m,1H),4.45(ddd,J=8.7,7.5,6.3Hz,1H),3.85(dt,J=8.1,2.2Hz,1H),3.48(ddd,J=10.9,4.5,2.1Hz,1H),3.39(t,J=11.0Hz,1H),2.50–2.33(m,1H),2.32–2.21(m,1H),1.11(s,3H),0.98(d,J=7.0Hz,3H),0.81(s,2H),0.79(d,J=6.3Hz,3H).13C NMR(150MHz,CDCl3)δ165.38,148.50,142.12,141.68,140.59,128.63(C×2),126.60,124.20(C×2),122.82,109.29,81.04,74.10,72.96,66.89,61.56,55.45,48.00,41.68,40.83,40.67,39.56,37.65,36.73,36.64,34.27,31.41,30.31,28.80,27.75,20.87,19.16,17.16,16.26,14.57.
(4)制备DG13-16:
称取150mg(0.349mmol,1equiv)7β-羟基薯蓣皂苷元置于25ml茄形瓶中,在依次加入85.2mg(0.698mmol,2equiv)DMAP,133.3mg(0.698mmol,2equiv)EDCI,169.6mg(1.047mmol,3equiv)α-甲基肉桂酸/202.1mg(1.047mmol,3equiv)2-硝基肉桂酸/184.5mg(1.047mmol,3equiv)对甲酰基肉桂酸后,加入10ml无水二氯甲烷进行充分溶解,于60℃中加热搅拌反应12h,薄层色谱法检测合成情况,合成结束后,取反应物在冰浴下缓慢滴加适量的4℃预冷的饱和碳酸氢钠,滴加完成后撤掉冰浴在常温下搅拌30min,用水和二氯甲烷萃取反应物,连续萃取3遍,合并下层有机相,将有机层与无水硫酸钠混合干燥,过滤,减压回收溶剂后,通过硅胶柱层析(石油醚∶乙酸乙酯)进行分离纯化,分别得到白色化合物,产率为40%35%/35%。
DG13:HR-ESI-MS m/z:575.3762[M+H]+(分子式经计算为C37H51O5,575.3736)。1HNMR(600MHz,CDCl3)δ7.67(d,J=1.6Hz,1H),7.39(d,J=5.0Hz,4H),4.85–4.66(m,1H),4.46(ddd,J=8.7,7.5,6.3Hz,1H),3.86(dt,J=8.2,2.2Hz,1H),3.48(ddd,J=10.9,4.5,2.1Hz,1H),3.39(t,J=11.0Hz,1H),2.51–2.36(m,2H),2.29(ddd,J=12.2,7.6,5.8Hz,1H),2.11(d,J=1.5Hz,3H),1.12(s,4H),0.98(d,J=6.9Hz,3H),0.81(s,3H),0.80(d,2H).13CNMR(150MHz,CDCl3)δ168.03,142.46,138.69,135.97,129.65(C×2),128.80,128.37,128.27,126.35,109.28,81.06,73.87,73.02,66.89,61.57,55.46,48.01,41.68,40.84,40.69,39.59,37.74,36.77,36.72,34.29,31.42,30.32,28.81,27.80,20.88,19.21,17.16,16.27,14.58,14.10
DG14:HR-ESI-MSm/z:781.3673[M+H]+(分子式经计算为C45H53N2O10,781.3700)。1HNMR(600MHz,CDCl3)δ8.24–7.92(m,4H),7.75–7.59(m,4H),7.61–7.37(m,2H),6.35(dd,J=15.8,12.1Hz,2H),5.38(t,J=1.8Hz,1H),5.19(dt,J=8.8,2.0Hz,1H),4.83–4.71(m,1H),4.39(ddd,J=8.7,7.5,6.3Hz,1H),3.47–3.42(m,1H),3.33(t,J=11.0Hz,1H),2.44(ddd,J=8.4,3.5,1.8Hz,2H),1.17(s,3H),0.97(d,J=7.0Hz,3H),0.83(s,3H),0.75(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ165.75,165.00,148.43,148.35,144.46,139.94,139.84,133.50,133.43,130.67,130.49,130.30,130.25,129.17,129.13,124.95,124.92,123.60,123.58,122.03,109.27,80.91,76.04,73.76,66.84,61.40,55.12,47.99,41.66,40.73,39.42,37.57,36.75,36.54,36.38,33.26,31.31,30.27,28.82,27.67,20.91,19.08,17.11,16.23,14.52.
DG15:HR-ESI-MSm/z:606.3370[M+H]+(分子式经计算为C36H48NO7,606.3431)。1HNMR(600MHz,CDCl3)δ8.14–7.94(m,2H),7.72–7.62(m,2H),7.54(ddd,J=8.7,6.2,2.6Hz,1H),6.34(d,J=15.8Hz,1H),5.35(t,J=2.0Hz,1H),4.86–4.66(m,1H),4.45(ddd,J=8.7,7.5,6.4Hz,1H),3.47(ddd,J=10.9,4.4,2.1Hz,1H),3.39(t,J=11.0Hz,1H),2.49–2.38(m,2H),2.28(ddd,J=12.2,7.5,5.8Hz,1H),1.11(s,3H),0.98(d,J=7.0Hz,3H),0.81(s,3H),0.79(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ165.13,148.36,142.23,139.88,133.51,130.66,130.27,129.15,126.52,124.94,123.59,109.28,81.06,74.02,72.99,66.88,61.56,55.46,48.00,41.68,40.83,40.68,39.57,37.64,36.74,36.66,34.28,31.41,30.32,28.81,27.74,20.87,19.17,17.16,16.27,14.58.
DG16:HR-ESI-MSm/z:747.3898[M+H]+(分子式经计算为C47H55O8,747.3897)。1HNMR(600MHz,CDCl3)δ8.09(d,J=15.8Hz,1H),8.06–8.01(m,1H),7.67–7.61(m,2H),7.54(ddd,J=8.7,6.2,2.6Hz,1H),6.34(d,J=15.8Hz,1H),5.35(t,J=2.0Hz,1H),4.83–4.67(m,1H),4.45(ddd,J=8.7,7.5,6.4Hz,1H),4.12(q,J=7.2Hz,0H),3.85(dt,J=8.1,2.2Hz,1H),3.49–3.44(m,1H),3.39(t,J=11.0Hz,1H),2.50–2.34(m,2H),2.28(ddd,J=12.2,7.5,5.8Hz,1H),1.11(s,3H),0.98(d,J=7.0Hz,3H),0.81(s,3H),0.79(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ191.50,191.44,166.33,165.59,144.48,143.14,142.94,140.12,140.07,137.18,137.16,130.18(C×4),128.61(C×2),128.50(C×2),122.08,121.68,121.64,109.30,80.89,75.87,73.62,66.85,61.41,55.14,48.00,41.67,40.74,39.41,37.64,36.77,36.55,36.41,33.27,31.49(d,J=10.8Hz),30.20(d,J=16.3Hz),28.80,27.70,20.90,19.09,17.09,16.24,14.53.
(5)制备DG17-18:
称取500mg(1.208mmol,1equiv)7β-羟基薯蓣皂苷元置于50ml茄形瓶中,在依次加入602.13mg(3.624mmol,3equiv)对氟肉桂酸,284.75mg(2.416mmol,2equiv)DMAP,480.78mg(2.416mmol,2equiv)EDCI后,加入20ml无水二氯甲烷进行充分溶解,60℃加热搅拌18h,薄层色谱法检测合成情况,合成结束后,取反应物在冰浴下缓慢滴加适量的4℃预冷的饱和碳酸氢钠,滴加完成后撤掉冰浴在常温下搅拌30min,用水和二氯甲烷萃取反应物,连续萃取3遍,合并下层有机相,将有机层与无水硫酸钠混合干燥,过滤,减压回收溶剂后,通过硅胶柱层析(石油醚∶乙酸乙酯)进行分离纯化,产率为40%/40%。
DG17:HR-ESI-MS m/z:726.3643[M+H]+(分子式经计算为C45H52F2O6,726.3732)。1HNMR(600MHz,CDCl3 0)δ7.63(dd,J=16.0,7.2Hz,2H),7.50(tdd,J=7.5,5.3,2.1Hz,4H),7.07(q,J=8.4Hz,4H),6.33(dd,J=16.0,2.8Hz,2H),5.35(t,J=1.8Hz,1H),5.18(dt,J=8.7,2.2Hz,1H),4.81–4.66(m,1H),4.44–4.29(m,1H),3.44(ddd,J=10.9,4.3,2.1Hz,1H),3.31(t,J=11.0Hz,1H),2.48–2.30(m,2H),1.16(s,3H),0.97(d,J=7.0Hz,3H),0.84(s,3H),0.75(d,J=6.3Hz,3H).13C NMR(151MHz,CDCl3)δ166.82,166.09,164.74,164.71,163.07,163.04,144.33,143.57,143.35,130.71(d,J=3.2Hz),130.66(d,J=4.0Hz),130.03(d,J=8.0Hz),129.91(d,J=8.6Hz),122.22,118.24,118.20,116.11(C×2),115.97(C×2),109.26,80.92,75.53,73.31,66.82,61.42,55.14,48.00,41.66,40.73,39.42,37.67,36.76,36.59,36.42,33.27,31.42(d,J=29.7Hz),30.21(d,J=17.4Hz),28.81,27.75,20.90,19.09,17.09,16.24,14.54.
DG18:HR-ESI-MS m/z:579.3476[M+H]+(分子式经计算为C36H48FO5,579.3486)。1HNMR(600MHz,CDCl3)δ7.63(d,J=15.9Hz,1H),7.54–7.45(m,2H),7.07(t,J=8.6Hz,2H),6.34(d,J=16.0Hz,1H),5.34(d,J=2.0Hz,1H),4.83–4.62(m,1H),4.45(ddd,J=8.7,7.4,6.3Hz,1H),3.85(dt,J=8.3,2.3Hz,1H),3.47(ddd,J=11.0,4.4,2.0Hz,1H),3.38(t,J=11.0Hz,1H),2.56–2.35(m,2H),2.28(ddd,J=12.8,7.5,5.7Hz,1H),1.11(s,3H),0.98(d,J=7.0Hz,3H),0.81(s,3H),0.79(d,J=6.4Hz,3H).13C NMR(150MHz,CDCl3)δ166.22,164.71,163.05,143.35,142.31,130.70(d,J=3.3Hz),129.92(d,J=8.5Hz),126.45,118.24,116.13,115.98,109.28,81.06,73.56,72.98,66.88,61.56,55.46,48.01,41.68,40.82,40.68,39.58,37.72,36.74,36.69,34.28,31.47(d,J=16.9Hz),30.23(d,J=26.1Hz),28.81,27.80,20.87,19.17,17.16,16.27,14.58.
对分离纯化得到的DG1-DG18采用1H NMR和13C NMR进行表征,得到NMR图谱,采用UPLC-Q-TOF-MS表征得到HR-ESI-MS图。结果见图1-54。
以下通过实验例证明本发明的有益结果。
实验例1、化合物对损伤神经细胞的保护活性测试
1.实验方法
本实验例采用CCK-8法检测细胞的存活率,具体方法如下:
人神经母细胞瘤SH-SY5Y细胞在含有10%澳洲胎牛血清的MEM/F12完全培养基中培养,利用H2O2造成SH-SY5Y细胞损伤模型。具体操作如下:取对数生长期的SH-SY5Y细胞消化离心收集,将细胞浓度调制5×105/ml,每孔100μL接种至96孔板中,于37℃、5%CO2培养箱中培养。待细胞贴壁生长后,弃去培养液,更换为新鲜的含药或不含药的培养基(细胞分组为:对照组(不添加药物的细胞组)、模型组、薯蓣皂苷元给药组、薯蓣皂苷元衍生物给药组)培养2h后,再与H2O2共同孵育24h,给药造模干预结束后,更换为含10% CCK-8溶液的培养基,培养4h后于酶标仪450nm波长下测定各孔光密度值(OD值),以空白组(纯PBS组)的OD值为参照来计算药物干预后SH-SY5Y细胞的相对存活率。
相对存活率计算公式为:相对存活率=[(给药组-空白组)/(对照组-空白组)]×100%。
相对保护率计算公式为:相对保护率=[(给药组-模型组)/(对照组-模型组)]×100%。
根据相对保护率来评价薯蓣皂苷元衍生物对SH-SY5Y细胞的保护作用。
本实验使用SPSS软件计算薯蓣皂苷元(DG)及其衍生物对SH-SY5Y细胞的在10μM下的细胞毒性以及药效,并利用GraphPad Prism 8.0.1制作统计柱形图。
2.实验结果
各化合物对SH-SY5Y细胞的毒性作用如图55所示,可以看出大部分DG1-DG18对于SH-SY5Y细胞表现出无毒作用。
各化合物对于过氧化氢损伤的SH-SY5Y细胞的保护作用结果如表2所示,可以看出,大部分薯蓣皂苷元衍生物对于过氧化氢损伤的SH-SY5Y细胞显示出保护作用,其中DG2、DG3、DG4、DG6、DG7、DG10-DG18的保护作用优于薯蓣皂苷元,衍生物DG6、DG12、DG13、DG15的保护作用甚至显著优于薯蓣皂苷元。
表2.薯蓣皂苷元及其衍生物对SH-SY5Y细胞的保护作用
化合物 | 相对保护率(%) | 化合物 | 相对保护率(%) |
DG | 21.62±3.19 | 10 | 26.41±2.77 |
DG1 | 21.16±2.79 | 11 | 29.28±1.98 |
DG2 | 26.41±2.77 | 12 | ** |
DG3 | 26.97±0.72 | 13 | ** |
DG4 | 30.71±0.95 | 14 | 28.64±2.52 |
DG5 | 21.52±2.78 | 15 | ** |
DG6 | ** | 16 | 26.48±0.52 |
DG7 | 27.92±2.50 | 17 | 29.41±6.16 |
DG8 | 21.19±3.16 | 18 | 29.88±2.76 |
DG9 | 21.16±2.79 |
注:**指与DG相比P<0.01。
上述实验结果表明,本发明提供的薯蓣皂苷元衍生物具有优良的神经保护作用,并且,其中DG2、DG3、DG4、DG6、DG7、DG10-DG18的保护作用优于薯蓣皂苷元。
综上,本发明提供了一种薯蓣皂苷元衍生物及其制备方法与用途。该薯蓣皂苷元衍生物对于过氧化氢损伤的SH-SY5Y细胞具有优良的保护作用,说明该薯蓣皂苷元衍生物具有优良的神经保护作用,并且,其中DG2、DG3、DG4、DG6、DG7、DG10-DG18的保护作用优于薯蓣皂苷元。本发明提供的薯蓣皂苷元衍生物的合成方法简单高效,成本低,在制备具有神经保护作用的药物中具有广阔的应用前景。
Claims (11)
1.一种化合物、其盐或其立体异构体,其特征在于:所述化合物的结构如式II所示:
其中,R1为氢或R2;
R2为
m选自0-5的整数;
R3各自独立地选自氢、C1-5烷基、卤素取代的C1-5烷基、硝基、卤素、-CHO;
R4为氢或C1-5烷基;
R5为氢或C1-5烷基。
2.根据权利要求1所述的化合物、其盐或其立体异构体,其特征在于:所述化合物的结构如式III或式IV所示:
其中,R2为
m选自0、1、2或3;
R3各自独立地选自氢、C1-3烷基、卤素取代的C1-3烷基、硝基、卤素、-CHO;
R4为氢或C1-3烷基。
3.根据权利要求2所述的化合物、其盐或其立体异构体,其特征在于:R2为
所述卤素为F、Cl或Br;
所述卤素取代的C1-3烷基为三氟甲基。
4.根据权利要求3所述的化合物、其盐或其立体异构体,其特征在于:R2选自以下基团:
5.根据权利要求1-4任一项所述的化合物、其盐或其立体异构体,其特征在于:所述化合物选自:
6.一种制备权利要求1-5任一项所述化合物的方法,其特征在于:所述方法包括以下步骤:
式I-a所示化合物与式I-b所示化合物反应,得到式II所示化合物;
其中,R1、R2、R3、R4、R5、m如权利要求1-5任一项所述。
7.根据权利要求6所述的方法,其特征在于:所述式I-a所示化合物的制备方法包括以下步骤:薯蓣皂苷元为底物与巨大芽孢杆菌共培养,发酵结束后收集发酵上清,提纯,得到式I-a所示化合物。
8.一种具有神经保护作用的药物,其特征在于:它是以权利要求1-5任一项所述化合物、其盐或其立体异构体为活性成分,加上药学上可接受的辅料制备得到的制剂。
9.权利要求1-5任一项所述化合物、其盐或其立体异构体在制备具有神经保护作用的药物中的用途。
10.根据权利要求9所述的用途,其特征在于:所述药物是预防和/或治疗神经退行性疾病的药物。
11.根据权利要求10所述的用途,其特征在于:所述神经退行性疾病为阿尔兹海默症、帕金森。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211393095.9A CN115746079B (zh) | 2022-11-08 | 2022-11-08 | 一种薯蓣皂苷元衍生物及其制备方法与用途 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211393095.9A CN115746079B (zh) | 2022-11-08 | 2022-11-08 | 一种薯蓣皂苷元衍生物及其制备方法与用途 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN115746079A CN115746079A (zh) | 2023-03-07 |
CN115746079B true CN115746079B (zh) | 2024-03-29 |
Family
ID=85368153
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211393095.9A Active CN115746079B (zh) | 2022-11-08 | 2022-11-08 | 一种薯蓣皂苷元衍生物及其制备方法与用途 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN115746079B (zh) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104250282A (zh) * | 2014-06-09 | 2014-12-31 | 吉林农业大学 | 薯蓣皂苷元氨基酸衍生物及其在制备抗肿瘤药物中的应用 |
CN109206472A (zh) * | 2018-09-27 | 2019-01-15 | 华东理工大学 | 薯蓣皂苷元衍生物、其药物组合物及其应用 |
-
2022
- 2022-11-08 CN CN202211393095.9A patent/CN115746079B/zh active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104250282A (zh) * | 2014-06-09 | 2014-12-31 | 吉林农业大学 | 薯蓣皂苷元氨基酸衍生物及其在制备抗肿瘤药物中的应用 |
CN109206472A (zh) * | 2018-09-27 | 2019-01-15 | 华东理工大学 | 薯蓣皂苷元衍生物、其药物组合物及其应用 |
Non-Patent Citations (1)
Title |
---|
Design, Synthesis and Biological Evaluation of Diosgenin-Amino Acid Derivatives with Dual Functions of Neuroprotection and Angiogenesis;Desheng Cai 等;《molecules》;第24卷;第4025(1-19)页 * |
Also Published As
Publication number | Publication date |
---|---|
CN115746079A (zh) | 2023-03-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP2019506133A (ja) | 微細藻類からフコキサンチンおよび/またはポリサッカライドを生産するための改善されたプロセス | |
CN108947949B (zh) | 抗焦虑氘代化合物及其医药用途 | |
CN108640968A (zh) | 一种混源萜类化合物及其在制备抗炎药物中的用途 | |
CN102766184A (zh) | 原人参二醇过氧化衍生物及其制备方法与应用 | |
CN113004297B (zh) | 二萜生物碱类化合物及其提取方法和应用 | |
CN104327147B (zh) | 一种麦角甾醇类化合物及其制备方法与应用 | |
CN115746079B (zh) | 一种薯蓣皂苷元衍生物及其制备方法与用途 | |
WO2013113294A1 (zh) | 一种甾醇类衍生物及其制备方法与应用 | |
US20210024570A1 (en) | Sterol derivatives and preparation method and uses thereof | |
KR101653586B1 (ko) | 프룩토실화 만지페린 및 이의 제조 방법 및 이의 용도 | |
CN110157764A (zh) | 一种地塞米松中间体的制备方法 | |
CN105085267A (zh) | 丹酚酸a的合成方法 | |
CN102718657A (zh) | 一种异甜菊醇化合物及其制备与应用 | |
CN108727403B (zh) | Nodosin衍生物及其制备方法和应用 | |
CN106536468B (zh) | 甘草素前驱物质的制备方法 | |
CN104224796A (zh) | 齐墩果烷型三萜类酯衍生物抗神经退行性药物用途 | |
CN112899191B (zh) | 一种诱变菌种以及从植物甾醇制备胆酸类化合物的方法 | |
CN111039913A (zh) | 一种具有抗肿瘤作用的化合物的制备方法 | |
CN114539282B (zh) | 一种具有抗骨质疏松功效的化合物及其制备方法和应用 | |
CN114573537B (zh) | 具有抗神经退行性改变活性的化合物、制备方法及其应用 | |
CN112301063B (zh) | 基于微生物转化续随子二萜烷型衍生物的方法及其制药用途 | |
US11312687B2 (en) | 7H-azulene [1,2,3-i,j] isoquinolin-7-one compound, single crystal and use thereof | |
CN116462727A (zh) | 一种β-烟酰胺核糖氯化物的制备方法 | |
CN116217642A (zh) | 木樨榄苷类化合物及其在制备抗肿瘤药物中的应用 | |
CN103725743A (zh) | 科罗索酸的生物催化合成方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |