CN116554240A - 关苍术中一种倍半萜乙酰糖苷及其制备方法与应用 - Google Patents
关苍术中一种倍半萜乙酰糖苷及其制备方法与应用 Download PDFInfo
- Publication number
- CN116554240A CN116554240A CN202310575953.XA CN202310575953A CN116554240A CN 116554240 A CN116554240 A CN 116554240A CN 202310575953 A CN202310575953 A CN 202310575953A CN 116554240 A CN116554240 A CN 116554240A
- Authority
- CN
- China
- Prior art keywords
- methanol
- volume ratio
- compound
- mixed solution
- silica gel
- 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.)
- Pending
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 229930004725 sesquiterpene Natural products 0.000 title claims abstract description 8
- 150000004354 sesquiterpene derivatives Chemical class 0.000 title claims abstract description 7
- 150000001875 compounds Chemical class 0.000 claims abstract description 33
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000000741 silica gel Substances 0.000 claims abstract description 16
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 16
- 239000003814 drug Substances 0.000 claims abstract description 13
- 206010006187 Breast cancer Diseases 0.000 claims abstract description 7
- 208000026310 Breast neoplasm Diseases 0.000 claims abstract description 7
- 206010009944 Colon cancer Diseases 0.000 claims abstract description 7
- 208000029742 colonic neoplasm Diseases 0.000 claims abstract description 7
- 239000011347 resin Substances 0.000 claims abstract description 7
- 229920005989 resin Polymers 0.000 claims abstract description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 87
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 45
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 27
- 239000011259 mixed solution Substances 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 239000002904 solvent Substances 0.000 claims description 9
- 238000004440 column chromatography Methods 0.000 claims description 8
- 238000010828 elution Methods 0.000 claims description 7
- 238000002953 preparative HPLC Methods 0.000 claims description 7
- 239000002994 raw material Substances 0.000 claims description 7
- 238000000605 extraction Methods 0.000 claims description 6
- 238000010898 silica gel chromatography Methods 0.000 claims description 6
- 241000132011 Atractylodes lancea Species 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 239000000243 solution Substances 0.000 claims description 5
- -1 guaiane glycoside Chemical class 0.000 claims description 4
- 230000002265 prevention Effects 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 3
- 239000003480 eluent Substances 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 229930182470 glycoside Natural products 0.000 claims description 3
- 238000000746 purification Methods 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 238000004809 thin layer chromatography Methods 0.000 claims description 3
- MCNAURNYDFSEML-UHFFFAOYSA-N Guaiane Natural products CC1CCC(C(C)=C)C(O)C2=C(C)C(=O)CC12 MCNAURNYDFSEML-UHFFFAOYSA-N 0.000 claims description 2
- 230000009897 systematic effect Effects 0.000 claims description 2
- FBMORZZOJSDNRQ-GLQYFDAESA-N Atractylenolide III Chemical compound C=C([C@@H]1C2)CCC[C@]1(C)C[C@@]1(O)C2=C(C)C(=O)O1 FBMORZZOJSDNRQ-GLQYFDAESA-N 0.000 claims 1
- 238000004128 high performance liquid chromatography Methods 0.000 claims 1
- 206010028980 Neoplasm Diseases 0.000 abstract description 10
- 229940079593 drug Drugs 0.000 abstract description 8
- 230000000694 effects Effects 0.000 abstract description 8
- 201000011510 cancer Diseases 0.000 abstract description 7
- 230000005764 inhibitory process Effects 0.000 abstract description 4
- 230000001093 anti-cancer Effects 0.000 abstract description 2
- 238000000338 in vitro Methods 0.000 abstract description 2
- 239000002246 antineoplastic agent Substances 0.000 abstract 1
- 229940041181 antineoplastic drug Drugs 0.000 abstract 1
- 238000004587 chromatography analysis Methods 0.000 abstract 1
- 230000009982 effect on human Effects 0.000 abstract 1
- 238000005516 engineering process Methods 0.000 abstract 1
- 238000001228 spectrum Methods 0.000 description 7
- 229910052799 carbon Inorganic materials 0.000 description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 5
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 238000005100 correlation spectroscopy Methods 0.000 description 3
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- XILIYVSXLSWUAI-UHFFFAOYSA-N 2-(diethylamino)ethyl n'-phenylcarbamimidothioate;dihydrobromide Chemical compound Br.Br.CCN(CC)CCSC(N)=NC1=CC=CC=C1 XILIYVSXLSWUAI-UHFFFAOYSA-N 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000002072 distortionless enhancement with polarization transfer spectrum Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 150000002338 glycosides Chemical class 0.000 description 2
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000007721 medicinal effect Effects 0.000 description 2
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- JUJWROOIHBZHMG-RALIUCGRSA-N pyridine-d5 Chemical compound [2H]C1=NC([2H])=C([2H])C([2H])=C1[2H] JUJWROOIHBZHMG-RALIUCGRSA-N 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- KWDNQVRRYNIDTM-UHFFFAOYSA-N (1S,4S,5S,7R,10R)-10,11,14-Trihydroxyguai-3-one 11-O-D-glucopyranoside Natural products C1CC(O)(CO)C2CC(=O)C(C)C2CC1C(C)(C)OC1OC(CO)C(O)C(O)C1O KWDNQVRRYNIDTM-UHFFFAOYSA-N 0.000 description 1
- 241000208838 Asteraceae Species 0.000 description 1
- 241000132003 Atractylis Species 0.000 description 1
- 241000132012 Atractylodes Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- VGAJNILINWUWOP-UHFFFAOYSA-N Eudesmane Natural products COC(=O)C(=C)C1C(O)C2C(=O)CCC(O)C2(C)CC1OC(=O)C(=C)CO VGAJNILINWUWOP-UHFFFAOYSA-N 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
- 241000147041 Guaiacum officinale Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical group CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 241000475481 Nebula Species 0.000 description 1
- 208000001140 Night Blindness Diseases 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 244000184734 Pyrus japonica Species 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 244000284012 Vetiveria zizanioides Species 0.000 description 1
- 235000007769 Vetiveria zizanioides Nutrition 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 206010061428 decreased appetite Diseases 0.000 description 1
- VILAVOFMIJHSJA-UHFFFAOYSA-N dicarbon monoxide Chemical group [C]=C=O VILAVOFMIJHSJA-UHFFFAOYSA-N 0.000 description 1
- 201000006549 dyspepsia Diseases 0.000 description 1
- LKBQARGGDFBGFF-UHFFFAOYSA-N eremophilane Natural products CC1C(O)CCC2C(=O)CC(CC12C)C(=C)C LKBQARGGDFBGFF-UHFFFAOYSA-N 0.000 description 1
- AJWBFJHTFGRNDG-GBJTYRQASA-N eremophilane Chemical compound C1CC[C@H](C)[C@@]2(C)C[C@H](C(C)C)CC[C@H]21 AJWBFJHTFGRNDG-GBJTYRQASA-N 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- DYEQPYSFRWUNNV-APIJFGDWSA-N eudesmane Chemical compound C1CC[C@@H](C)[C@@H]2C[C@H](C(C)C)CC[C@]21C DYEQPYSFRWUNNV-APIJFGDWSA-N 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000004438 eyesight Effects 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 150000008131 glucosides Chemical class 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 229940091561 guaiac Drugs 0.000 description 1
- QAQCPAHQVOKALN-RMEBNNNOSA-N guaiane Chemical compound C1[C@H](C(C)C)CC[C@H](C)[C@@H]2CC[C@H](C)[C@@H]21 QAQCPAHQVOKALN-RMEBNNNOSA-N 0.000 description 1
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 1
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 1
- SNVLJLYUUXKWOJ-UHFFFAOYSA-N methylidenecarbene Chemical compound C=[C] SNVLJLYUUXKWOJ-UHFFFAOYSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000013307 optical fiber Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Substances [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 210000004243 sweat Anatomy 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/18—Acyclic radicals, substituted by carbocyclic rings
-
- 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
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Steroid Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
关苍术中一种倍半萜乙酰糖苷及其制备方法与应用。本发明属于医药技术领域,涉及利用大孔树脂、硅胶及制备色谱等技术从关苍术根茎中提取分离一种倍半萜乙酰糖苷化合物关苍术愈创木烷苷I,制备方法简单易行。该化合物分子式为C23H38O10,体外人源癌细胞抑制活性研究表明其具有抗癌活性,包括对人结肠癌细胞HT‑29、人乳腺癌MCF‑7细胞均具有明显的抑制作用,有望开发为新的抗癌药物。
Description
技术领域
本发明具体涉及一种具有癌细胞抑制作用的倍半萜乙酰糖苷类化合物。
背景技术
关苍术(Atractylodes japonica Koidz.ex Kitam.)为菊科苍术属多年生草本植物,其药用部位为根茎,味辛、苦,性温无毒。主要产自我国东北三省。自2005年起,关苍术药材已是吉林省(道地产区)药材品种的重点保护对象。历代医学著作对其药用价值的评价都很高,其药用部位根茎具有健脾燥湿、发汗解表明目、祛风等功效,主要用于治疗食欲不振,消化不良;内外翳障,夜盲症等病症。关苍术与苍术的主要化学成分及功效相似,且关苍术在东北拥有丰富的野生资源,为了资源利用更加合理,将其开发成苍术替代品,充分开发其药用价值。
倍半萜类化合物是关苍术的主要化学成分类型之一,从结构母核分类主要包括桉叶烷型、愈创木烷型、香根螺烷型和艾里莫份烷型4种,它们多数具有广泛的生物活性,包括抗癌、抗炎、抗菌、神经保护、保肝等活性,具有很好的开发前景和价值。
发明内容
本发明的目的是在于提供一种新的倍半萜乙酰糖苷类化合物及其制备方法和医疗用途,以期扩大治疗癌症药物的资源、来源;同时为了提高关苍术的资源开发与利用价值。
为了达到上述目的,本发明从关苍术中提取分离得到一种具有抗结肠癌和乳腺癌活性的倍半萜类化合物关苍术愈创木烷苷I,其结构式如下:
。
本发明还提供了关苍术愈创木烷苷I的制备方法:以关苍术为原料,依次通过醇提、柱层析制备获得。
上述柱层析依次包括大孔树脂柱、正相硅胶柱、反相硅胶柱和制备型HPLC。
本发明化合物关苍术愈创木烷苷I的具体制备步骤如下:
(1)醇提:以关苍术干燥根茎为原料,适当粉碎,采用70%乙醇回流提取3次,过滤,合并3次滤液,减压回收溶剂,干燥后得浸膏状提取物;
(2)富集纯化:将步骤(1)所得浸膏状提取物用水分散至相对密度为1.25±0.05g/mL的溶液,经AB-8型大孔树脂柱色谱富集纯化,分别以水、30%乙醇、50%乙醇、95%乙醇依次洗脱,收集50%乙醇洗脱液,减压回收溶剂得50%乙醇洗脱部分;
(3)正相硅胶柱层析:取步骤(2)所得50%乙醇洗脱部分采用正相硅胶柱,依次采用体积比为8:1的二氯甲烷和甲醇混合溶液、体积比为5:1的二氯甲烷和甲醇混合溶液、体积比为3:1的二氯甲烷和甲醇混合溶液进行系统梯度洗脱,收集体积比为3:1的馏分液,减压回收溶剂;
(4)反相硅胶柱层析:取步骤(3)制备的馏分部位,通过反相硅胶ODS柱色谱,依次以体积比为1:2的甲醇和水混合溶液、体积比为1:1的甲醇和水混合溶液、体积比为2:1的甲醇和水混合溶液进行洗脱,收集体积比为1:1的甲醇和水混合溶液洗脱部分,经反相硅胶薄层色谱层析检识后相近者合并,依次得到Fr.1、F.2、Fr.3三部分;
(5)制备型HPLC纯化:将步骤(4)所得Fr.2部分采用甲醇溶解进入制备型HPLC,流动相为体积比为36:64的甲醇和水混合溶液,流速为3 mL/min,收集馏分即得本发明化合物。
本发明还提供了关苍术愈创木烷苷I在制备预防和治疗癌症药物方面的应用。优选为在制备预防和治疗结肠癌和乳腺癌药物方面的应用。
本发明相比现有技术具有以下优点:其一,本发明化合物具有制备临床癌症预防和治疗药物的前景,扩大了药物来源;其二,以关苍术为原料能通过多种药用途径开发替代濒危药材苍术。
附图说明
图1为本发明化合物的化学结构式;
图2为本发明化合物的正性HR-ESI-MS谱图;
图3为本发明化合物的1H-NMR谱图;
图4为本发明化合物的13C-NMR谱图;
图5为本发明化合物的DEPT谱图;
图6为本发明化合物的HSQC谱图;
图7为本发明化合物的HMBC谱图;
图8本发明化合物的1H-1H COSY谱图
图9为本发明化合物的NOESY谱图。
实施方式
根据本发明所公开的技术内容,本领域技术人员很清楚本发明的其他实施方案,下述实施方案仅作示例,在不违反本发明主旨及范围的情况下,可对本发明进行各种调整和改进,这些变化应在本发明的保护范围内,下面结合具体实施例对本发明进行详细说明。
实施案例1本发明化合物的制备方法:
(1)醇提:以关苍术干燥根茎10 kg为原料,适当粉碎成粗粉过1号筛,采用70%乙醇回流提取3次,每次2 h,其中药材重量与乙醇体积配比为1:6,提取结束过滤,合并3次滤液,减压回收溶剂,干燥后得浸膏状提取物2.4 kg;
(2)富集纯化:将步骤(1)所得浸膏状提取物用水分散至相对密度为1.25±0.05g/mL的溶液,经AB-8型大孔树脂柱色谱富集纯化(色谱柱内径10 cm,长1.80 m,其中树脂有效高度1.35 m),分别以水、30%乙醇、50%乙醇、95%乙醇依次洗脱3个柱体积、2个柱体积、3个柱体积、2个柱体积,收集50%乙醇洗脱液,减压回收溶剂得50%乙醇洗脱部分386 g;
(3)正相硅胶柱层析:取步骤(2)所得50%乙醇洗脱部分采用正相硅胶柱(色谱柱内径6.5 cm长1.8 m,其中硅胶有效高度1.2 m),依次采用体积比为8:1的二氯甲烷和甲醇混合溶液洗脱3柱体积、体积比为5:1的二氯甲烷和甲醇混合溶液洗脱2.5柱体积、体积比为3:1的二氯甲烷和甲醇混合溶液洗脱3个柱体积,收集体积比为3:1的馏分液,减压回收溶剂得收集馏分36g;
(4)反相硅胶柱层析:取步骤(3)制备的馏分部位,通过反相硅胶ODS柱色谱(色谱柱内径5 cm,长1.5 m,其中反相硅胶有效高度0.8 m),依次以体积比为1:2的甲醇和水混合溶液洗脱3个柱体积、体积比为1:1的甲醇和水混合溶液2个柱体积、体积比为2:1的甲醇和水混合溶液进行洗脱2.5个柱体积,收集体积比为1:1的甲醇和水混合溶液洗脱部分,经反相硅胶薄层色谱层析检识后相近者合并,依次得到Fr.1、F.2、Fr.3三部分;
(5)制备型HPLC纯化:将步骤(4)所得Fr.2部分采用甲醇溶解进入制备型HPLC(Waters,515-2414,SunFireTM Prep C18,250 mm×10 mm i.d.,5μm),进样浓度不超过35mg/mL,流动相为体积比为36:64的甲醇和水混合溶液,流速为3 mL/min,tR=26.5~26.9 min收集馏分后得到本发明化合物(8.8 mg)。
实施案例2本发明化合物的表征:
实施例1所得化合物为黄色油状物,溶于甲醇,Molish反应呈阳性,酸水解薄层检出D-葡萄糖,25℃下比旋光度为+12°(c=0.15 MeOH)。在HR-ESI-MS中,m/z 492.28027处可见[M+NH4]+离子峰(calcd for 492.2809),表明本发明化合物的分子量为474。结合1H-NMR、13C-NMR及DEPT谱等推测该化合物的分子式为C23H38O10,计算其不饱和度为5。
在1H-NMR (600 MHz, Pyridine-d 5 )谱中,见图3,观察到有4个甲基质子信号δ H1.41 (3H, s, H-12)、1.44 (3H, s, H-13)、1.15 (3H, d, J = 7.2 Hz, H-15)、2.02(3H, s, H-2''),5个亚甲基质子信号δ H 2.63 (1H, dd, J = 18.6, 9.0 Hz, H-2α)、3.09(1H, dd, J = 18.6, 10.2 Hz, H-2β)、1.24 (1H, m, H-6α)、2.49 (1H, m, H-6β)、1.63(1H, m, H-8α)、2.34 (1H, m, H-8β)、1.97 (1H, m, H-9α)、2.68 (1H, m, H-9β)、3.77(2H, br.s, H-14),2个次甲基质子信号δ H 1.89 (1H, m, H-4)、2.04 (1H, m, H-7),以及一组糖质子信号δ H 4.99 (1H, d, J = 7.2 Hz, H-1')、3.99 (1H, m, H-2')、4.20 (1H,m, H-3')、4.02 (1H, m, H-4')、3.97 (1H, m, H-5')、4.70 (1H, dd, J = 11.4, 6.6Hz, H-6'α)、4.91 (1H, d, J = 11.0 Hz, H-6'β),其中δ H 4.99 (1H, d, J = 7.2 Hz,H-1')为端基质子信号,且说明该糖为β构型。
在13C-NMR (150 MHz, Pyridine-d 5 )谱,见图4,观察到23个碳信号,其中1个羰基碳信号δ C 219.4,1个酯基碳信号δ C 170.7。结合DEPT谱,见图5,判断有4个甲基碳信号δ C24.2、24.2、12.8、20.8,6个亚甲基碳信号δ C 39.3、36.6、23.4、37.1、69.6、64.9,9个次甲基碳信号δ C 46.8、44.0、52.2、50.6、98.6、75.2、78.6、71.7、74.7,2个季碳信号δ C 74.4、80.5,其中δ C 98.6、75.2、78.6、71.7、74.7、64.9为6个糖上碳信号。
在1H-1H COSY谱中,见图8,H2-2与H-1相关,H-1与H-5相关,H-5与H-4、H2-6相关,H-4与H3-15,H2-6与H-7相关,H-7与H2-8相关,H2-8与H2-9相关,故推断母核结构中存在片段“-CH2-CH-CH-CH-CH3”和“-CH-CH2-CH-CH2-CH2-”,且在C-5上相连。此外,1H-1H COSY谱中还存在H-1'与H-2'相关,H-2'与H-3'相关,H-3'与H-4'相关,H-4'与H-5',可推断结构中存在片段“-CH-CH-CH-CH-”,通过1H-NMR、13C-NMR和HSQC谱,分别见图3、图4、图6,推断该片段为糖上部分。该化合物的母核结构与参考文献[Kitajima J,Kamoshit A,Ishikawa T,etal.Glycosides of Atractylodes Japonica.Chem Pharm Bull,2003,51(2): 152-157]中的化合物(1S,4S,5S,7R,10R)-10,11,14-trihydroxyguai-3-one 11-O-β-D-glucopyranoside结构相似,通过DEPT、HSQC和HMBC谱,分别见图5、图6、图7,推断出二者主要区别是糖上6'位的羟基氢被乙酰基取代。在NOESY谱中,见图9,H-1β与H-4、H-6 (δ H1.24)、H2-14的相关信号、H-5与Me-15的相关信号、H-12与H-6 (δ H 2.49)、H-7的相关信号表明H-4、C-10羟甲基部分、C-7二甲基甲醇部分为β构型,H-5、H-7、Me-15、OH-10为α构型。
综合上述结构解析,NMR主要数据如表1所示,SciFinder数据库检索确定为新化合物,命名为关苍术愈创木烷苷I,结构式如图1所示。
效果实施例
(1)材料和方法
对实施例1中制备得到的化合物进行了体外抗癌活性实验,实验所采用的细胞为人结肠癌HT-29细胞和人乳腺癌MCF-7细胞,上述瘤株均购自赛尔试剂公司,采用MTT法进行活性测试。
具体方法如下:
取对数生长期生产状态良好的HT-29、MCF-7细胞,调整各细胞密度至104个/孔接种于96孔板中,每孔为100 μL的细胞悬液。接种好后,将上述人源癌细胞置于含10% L-谷氨酰胺的胎牛血清的RPMI 1640培养液中(另含有100 μg/mL盘尼西林和100 μg/mL链霉素),然后放入恒温37℃,5%CO2培养箱中培养24 h后给药,待测不同浓度药物分别为5、10、20、40、80、160 μmol/L,另设空白孔和对照孔(空白孔未接种细胞,对照孔为不含药物的培养液),顺铂为阳性对照组,上述每组均设3个复孔,继续孵育48 h后,每孔加入5 mg/mL 的MTT20 μL,继续培养4 h后离心,吸弃96孔板中孔内培养上清液,再每孔加入150 μL DMSO,置摇床上低温振荡10 min,用酶联免疫检测仪在波长570 nm波长处测定每孔的吸光值,计算受试药物对癌细胞生长抑制率。细胞存活抑制率%=[1-(实验组A-空白组A)/(对照组A-空白组A)]×100%。使用Logit法计算药物的IC50值。结果如下表2所示:
(2)结果
经线性回归计算IC50值显示本发明涉及的倍半萜类化合物关苍术愈创木烷苷I对人结肠癌HT-29细胞和人乳腺癌MCF-7细胞作用IC50值分别为69.17±1.27 μmol/L和45.62±2.19 μmol/L,顺铂作为阳性对照药对人结肠癌HT-29细胞和人乳腺癌MCF-7细胞作用IC50值分别为7.52±0.83 μmol/L和5.16±0.64 μmol/L。
综上所述,本发明从关苍术根茎中分离得到的新化合物关苍术愈创木烷苷I具有制备临床癌症预防和治疗药物的前景。
Claims (3)
1.一种从关苍术根茎中分离得到的一种倍半萜乙酰糖苷类化合物,所述化合物的分子式为C23H38O10,命名为关苍术愈创木烷苷I,其结构式如下:
。
2.权利要求1所述化合物的制备方法,其特征在于:所述关苍术愈创木烷苷I以关苍术根茎为原料,大孔树脂柱、正相硅胶柱、反相硅胶柱和HPLC制备得到,具体制备步骤如下:
(1)醇提:以关苍术干燥根茎为原料,适当粉碎,采用70%乙醇回流提取3次,过滤,合并3次滤液,减压回收溶剂,干燥后得浸膏状提取物;
(2)富集纯化:将步骤(1)所得浸膏状提取物用水分散至相对密度为1.25±0.05 g/mL的溶液,经AB-8型大孔树脂柱色谱富集纯化,分别以水、30%乙醇、50%乙醇、95%乙醇依次洗脱,收集50%乙醇洗脱液,减压回收溶剂得50%乙醇洗脱部分;
(3)正相硅胶柱层析:取步骤(2)所得50%乙醇洗脱部分采用正相硅胶柱,依次采用体积比为8:1的二氯甲烷和甲醇混合溶液、体积比为5:1的二氯甲烷和甲醇混合溶液、体积比为3:1的二氯甲烷和甲醇混合溶液进行系统梯度洗脱,收集体积比为3:1的馏分液,减压回收溶剂;
(4)反相硅胶柱层析:取步骤(3)制备的馏分部位,通过反相硅胶ODS柱色谱,依次以体积比为1:2的甲醇和水混合溶液、体积比为1:1的甲醇和水混合溶液、体积比为2:1的甲醇和水混合溶液进行洗脱,收集体积比为1:1的甲醇和水混合溶液洗脱部分,经反相硅胶薄层色谱层析检识后相近者合并,依次得到Fr.1、F.2、Fr.3三部分;
(5)制备型HPLC纯化:将步骤(4)所得Fr.2部分采用甲醇溶解进入制备型HPLC,流动相为体积比为36:64的甲醇和水混合溶液,流速为3 mL/min,收集馏分即得本发明化合物。
3.权利要求1所述化合物关苍术愈创木烷苷I在制备预防和治疗结肠癌和乳腺癌药物方面的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310575953.XA CN116554240A (zh) | 2023-05-22 | 2023-05-22 | 关苍术中一种倍半萜乙酰糖苷及其制备方法与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310575953.XA CN116554240A (zh) | 2023-05-22 | 2023-05-22 | 关苍术中一种倍半萜乙酰糖苷及其制备方法与应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN116554240A true CN116554240A (zh) | 2023-08-08 |
Family
ID=87499888
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310575953.XA Pending CN116554240A (zh) | 2023-05-22 | 2023-05-22 | 关苍术中一种倍半萜乙酰糖苷及其制备方法与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116554240A (zh) |
-
2023
- 2023-05-22 CN CN202310575953.XA patent/CN116554240A/zh active Pending
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN112094176B (zh) | 一种从山橿中提取的二苯乙烯类化合物及其制备方法与应用 | |
CN110627861B (zh) | 一种知母甾体皂苷化合物及其制备方法和应用 | |
CN109879844A (zh) | 小花清风藤中七种黄酮类化学成分的提取分离方法 | |
CN110305014B (zh) | 环新木脂烷型木脂素对映异构体及其制备和应用 | |
CN110511194B (zh) | 四氢呋喃型木脂素化合物及其制备方法和应用 | |
CN110343045B (zh) | 芳基四氢萘型木脂素类化合物及制备和应用 | |
CN115991692B (zh) | 菘蓝中螺二烯酮木脂素化合物的制备方法及其应用 | |
CN113754620B (zh) | 火麻仁中的木脂素酰胺类化合物及其制备方法和应用 | |
CN114436802B (zh) | 一种杜松烷倍半萜化合物及其制备方法和应用 | |
CN109180471A (zh) | 水栀子单萜类化合物crocusatinN和jasminosideB制备方法及应用 | |
CN113666894B (zh) | 一种从老鹰茶中提取分离呋喃酮类化合物的方法及其应用 | |
CN116554240A (zh) | 关苍术中一种倍半萜乙酰糖苷及其制备方法与应用 | |
CN108129439A (zh) | 一种桃儿七中两个具有抗肿瘤活性的双黄酮类化合物的制备方法及其应用 | |
CN107325069B (zh) | 一种倍半萜类化合物的提取方法 | |
CN112409152A (zh) | 鲜地黄中艾里莫酚烷型地黄倍半萜a和地黄倍半萜b的制备方法及其应用 | |
CN109206392B (zh) | 一种香豆素类化合物及其制备方法与应用 | |
CN117624268A (zh) | 关苍术中一种环木脂素苷类化合物及其制备方法与应用 | |
CN116589514A (zh) | 关苍术中一种愈创木烷型倍半萜苷及其制备方法与应用 | |
CN115611844B (zh) | 一种从关苍术中分离得到的化合物的制备方法和应用 | |
CN114456137B (zh) | 螺环萘类化合物及其制备方法和应用 | |
CN109180696B (zh) | 环烯酮类化合物及其制备方法和应用 | |
CN113387995B (zh) | 一种从马桑狗帮中提取分离的三萜化合物及其制备方法和应用 | |
CN109897079B (zh) | 一种香豆素糖苷类化合物的制备方法及其应用 | |
CN109776565B (zh) | 一种苦味素类化合物及其制备方法与应用 | |
CN115651055B (zh) | 一种齐墩果烷型三萜皂苷化合物及其制备方法和应用 |
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 |