CN117756763A - 一种苯并吡喃对映异构体及其制备方法与应用 - Google Patents
一种苯并吡喃对映异构体及其制备方法与应用 Download PDFInfo
- Publication number
- CN117756763A CN117756763A CN202311704918.XA CN202311704918A CN117756763A CN 117756763 A CN117756763 A CN 117756763A CN 202311704918 A CN202311704918 A CN 202311704918A CN 117756763 A CN117756763 A CN 117756763A
- Authority
- CN
- China
- Prior art keywords
- benzopyran
- compound
- enantiomers
- enantiomer
- extract
- 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
- 150000001562 benzopyrans Chemical class 0.000 title claims abstract description 32
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 239000000284 extract Substances 0.000 claims abstract description 18
- 210000004185 liver Anatomy 0.000 claims abstract description 9
- KYNSBQPICQTCGU-UHFFFAOYSA-N Benzopyrane Chemical compound C1=CC=C2C=CCOC2=C1 KYNSBQPICQTCGU-UHFFFAOYSA-N 0.000 claims abstract description 8
- 230000002633 protecting effect Effects 0.000 claims abstract description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 27
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 24
- 229940125904 compound 1 Drugs 0.000 claims description 21
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 10
- HBENZIXOGRCSQN-VQWWACLZSA-N (1S,2S,6R,14R,15R,16R)-5-(cyclopropylmethyl)-16-[(2S)-2-hydroxy-3,3-dimethylpentan-2-yl]-15-methoxy-13-oxa-5-azahexacyclo[13.2.2.12,8.01,6.02,14.012,20]icosa-8(20),9,11-trien-11-ol Chemical compound N1([C@@H]2CC=3C4=C(C(=CC=3)O)O[C@H]3[C@@]5(OC)CC[C@@]2([C@@]43CC1)C[C@@H]5[C@](C)(O)C(C)(C)CC)CC1CC1 HBENZIXOGRCSQN-VQWWACLZSA-N 0.000 claims description 9
- GVOISEJVFFIGQE-YCZSINBZSA-N n-[(1r,2s,5r)-5-[methyl(propan-2-yl)amino]-2-[(3s)-2-oxo-3-[[6-(trifluoromethyl)quinazolin-4-yl]amino]pyrrolidin-1-yl]cyclohexyl]acetamide Chemical compound CC(=O)N[C@@H]1C[C@H](N(C)C(C)C)CC[C@@H]1N1C(=O)[C@@H](NC=2C3=CC(=CC=C3N=CN=2)C(F)(F)F)CC1 GVOISEJVFFIGQE-YCZSINBZSA-N 0.000 claims description 9
- 238000000926 separation method Methods 0.000 claims description 9
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 8
- 239000007924 injection Substances 0.000 claims description 8
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 238000002953 preparative HPLC Methods 0.000 claims description 7
- 239000000243 solution Substances 0.000 claims description 7
- 239000003814 drug Substances 0.000 claims description 6
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 5
- 238000007605 air drying Methods 0.000 claims description 4
- 239000002021 butanolic extract Substances 0.000 claims description 4
- 238000010829 isocratic elution Methods 0.000 claims description 4
- 239000003208 petroleum Substances 0.000 claims description 4
- 238000010298 pulverizing process Methods 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 4
- 238000000825 ultraviolet detection Methods 0.000 claims description 4
- HBAQYPYDRFILMT-UHFFFAOYSA-N 8-[3-(1-cyclopropylpyrazol-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one Chemical class C1(CC1)N1N=CC(=C1)C1=NNC2=C1N=C(N=C2)N1C2C(N(CC1CC2)C)=O HBAQYPYDRFILMT-UHFFFAOYSA-N 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims description 3
- 239000008194 pharmaceutical composition Substances 0.000 claims description 3
- 238000010898 silica gel chromatography Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 241000501743 Gentiana macrophylla Species 0.000 abstract description 13
- 230000000694 effects Effects 0.000 abstract description 8
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 abstract description 6
- 238000000338 in vitro Methods 0.000 abstract description 2
- 208000027418 Wounds and injury Diseases 0.000 abstract 1
- 230000006378 damage Effects 0.000 abstract 1
- 230000005764 inhibitory process Effects 0.000 abstract 1
- 208000014674 injury Diseases 0.000 abstract 1
- 210000005229 liver cell Anatomy 0.000 abstract 1
- 229960005489 paracetamol Drugs 0.000 abstract 1
- 150000001875 compounds Chemical class 0.000 description 6
- 238000001228 spectrum Methods 0.000 description 5
- PWKSKIMOESPYIA-BYPYZUCNSA-N L-N-acetyl-Cysteine Chemical compound CC(=O)N[C@@H](CS)C(O)=O PWKSKIMOESPYIA-BYPYZUCNSA-N 0.000 description 4
- 208000029618 autoimmune pulmonary alveolar proteinosis Diseases 0.000 description 4
- HIXDQWDOVZUNNA-UHFFFAOYSA-N 2-(3,4-dimethoxyphenyl)-5-hydroxy-7-methoxychromen-4-one Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(OC)C(OC)=C1 HIXDQWDOVZUNNA-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical group C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 3
- 230000005779 cell damage Effects 0.000 description 3
- 208000037887 cell injury Diseases 0.000 description 3
- 230000002596 correlated effect Effects 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
- 241001071795 Gentiana Species 0.000 description 2
- 206010023126 Jaundice Diseases 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 108010087230 Sincalide Proteins 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000010609 cell counting kit-8 assay Methods 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 231100000263 cytotoxicity test Toxicity 0.000 description 2
- 238000002212 electronic circular dichroism spectrum Methods 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 238000002156 mixing Methods 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
- 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
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000001026 1H--1H correlation spectroscopy Methods 0.000 description 1
- SAXKWTPDZMBKSQ-UHFFFAOYSA-N 2,2-dimethylchromene Chemical compound C1=CC=C2C=CC(C)(C)OC2=C1 SAXKWTPDZMBKSQ-UHFFFAOYSA-N 0.000 description 1
- 208000006820 Arthralgia Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000967672 Gentiana crassicaulis Species 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010062016 Immunosuppression Diseases 0.000 description 1
- ZUKLFFYDSALIQW-MSUKCBDUSA-N Iridoid glycoside Chemical compound [H][C@]12CC[C@H](C(O)=O)[C@@]1([H])[C@H](OC1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)OC=C2 ZUKLFFYDSALIQW-MSUKCBDUSA-N 0.000 description 1
- 206010023201 Joint contracture Diseases 0.000 description 1
- 206010033799 Paralysis 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
- 208000004880 Polyuria Diseases 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 229960004308 acetylcysteine Drugs 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- -1 benzopyran compound Chemical class 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 230000004531 blood pressure lowering effect Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000005100 correlation spectroscopy Methods 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000002784 cytotoxicity assay Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000012010 growth Effects 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
- 238000005570 heteronuclear single quantum coherence Methods 0.000 description 1
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 1
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 229930182489 iridoid glycoside Natural products 0.000 description 1
- 229930013686 lignan Natural products 0.000 description 1
- 150000005692 lignans Chemical class 0.000 description 1
- 235000009408 lignans Nutrition 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 231100001083 no cytotoxicity Toxicity 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 235000019633 pungent taste Nutrition 0.000 description 1
- 230000002040 relaxant effect Effects 0.000 description 1
- 238000001896 rotating frame Overhauser effect spectroscopy Methods 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 239000012224 working solution 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
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种苯并吡喃对映异构体及其制备方法与应用。本发明从秦艽根茎中提取分离得到一对苯并吡喃对映异构体,通过检测该苯并吡喃对映异构体对对乙酰氨基酚体外诱导人肝细胞(L‑O2)损伤的抑制作用,评价秦艽苯并吡喃的保肝活性。结果发现,此苯并吡喃对映异构体具有较好的保肝活性。本发明提供的苯并吡喃对映异构体的制备方法操作简单且成本低廉,得到的苯并吡喃纯度高,结构明确,具有多种潜在的应用价值,为开发秦艽苯并吡喃的药用价值奠定基础。
Description
技术领域
本发明属于天然产物化学技术领域,具体涉及一种苯并吡喃对映异构体及其制备方法与应用。
背景技术
秦艽Gentiana macrophylla Pall.为龙胆科龙胆属秦艽组多年生草本植物,始载于《神农本草经》,别名左秦艽、大叶龙胆。与麻花秦艽(Gentiana stramin ea Maxim.)、粗茎秦艽(Gentiana crassicaulis Duthie ex Burk.)、小秦艽(Gentian a dahuricaFisch.)统称“秦艽”。在新疆、宁夏、陕西、山西、河北等地均有种植,多生长于海拔400-2400m的河滩、山坡草地、草甸、林下及林缘,其味苦、辛,性平、微寒,归胃、肝、胆经,具有祛风湿、止痹痛、退虚热、舒筋络、利湿退黄的功效,常用于全身性风湿痛、周身或关节拘挛、手足不遂、骨蒸潮热、湿热黄疸等病症。秦艽所含环烯醚萜苷类是其主要活性成分,此外还含有木脂素、黄酮、生物碱、多糖等天然成分,具有抗炎镇痛、保肝、抗病毒、抗肿瘤、免疫抑制、降压等药理作用。目前,对中药秦艽的化学成分和生物活性报道多集中于环烯醚萜类上,其他成分如苯并吡喃类化合物报道较少。
发明内容
本发明的第一目的是提供一对苯并吡喃对映异构体,本发明的第二目的是提供所述苯并吡喃对映异构体的制备方法,本发明的第三目的是提供所述苯并吡喃对映异构体的应用。
本发明的第一目的是这样实现的,一对苯并吡喃对映异构体,该对映异构体具有式1a和式1b的结构:
本发明的第二目的是这样实现的,所述苯并吡喃对映异构体的制备方法,具体按以下步骤实现:
1)将秦艽药材洗净晾干,粉碎后用70%乙醇浸泡提取3-4次,每次24-48h,将提取液减压浓缩,得浸膏;浸膏加水搅匀,依次用石油醚,乙酸乙酯,正丁醇萃取,浓缩回收正丁醇萃取相得正丁醇相浸膏;
2)将正丁醇相浸膏进行硅胶柱层析,用体积比为80:1,50:1,30:1,15:1及0:1的二氯甲烷/甲醇进行梯度洗脱,收集体积比为15:1的洗脱液,减压浓缩,再通过半制备高效液相色谱,用Agilent Eclipse XDB-C18(9.4×250mm,5μm,3ml/min)分离,得到苯并吡喃类化合物1;
3)将得到的化合物1通过CHIRALPAK IH手性柱进行HPLC手性分离得到化合物1a和化合物1b。
步骤2)中,半制备高效液相色谱分离采用体积比为30:70的甲醇/水进行等度洗脱。
步骤3)中,HPLC手性分离采用体积比为50:50的正己烷/异丙醇进行等度洗脱,进样量为20μL,进样速度为1.5ml/min。
本发明的第三目的是这样实现的,所述苯并吡喃对映异构体的应用为在制备保肝药物中的应用。
本发明从秦艽根茎中提取分离得到一对苯并吡喃对映异构体,此苯并吡喃对映异构体具有较好的保肝活性。本发明提供的苯并吡喃对映异构体的制备方法操作简单且成本低廉,得到的苯并吡喃纯度高,结构明确,具有多种潜在的应用价值,为开发秦艽中苯并吡喃化合物的药用价值奠定基础。
附图说明
图1为本实施例1制备的化合物1的红外光谱图;
图2为实施例1制备的化合物1的核磁共振氢谱图;
图3为实施例1制备的化合物1的核磁共振碳谱图;
图4为实施例1制备的化合物1的COSY图;
图5为实施例1制备的化合物1的HSQC图;
图6为实施例1制备的化合物1的HMBC图;
图7为实施例1制备的化合物1的ROESY图;
图8为实施例1制备的化合物1a及化合物1b的ECD量子计算图;
图9中A、B、C图分别为本发明苯并吡喃化合物1、1a及1b对L-O2细胞毒性检测结果(NC组为正常组;**p<0.01与正常组比较);
图10中A、B、C图分别为本发明苯并吡喃化合物1、1a及1b对APAP诱导L-O2细胞损伤的影响(NC组为正常组,AP为模型组,NAC阳性组;#p<0.05,##p<0.01,###p<0.001和####p<0.0001与正常组比较;*p<0.05,**p<0.01,
***p<0.001和****p<0.0001与模型组比较)。
具体实施方式
下面对本发明作进一步的说明,但不以任何方式对本发明加以限制,基于本发明所作的任何变换,均属于本发明的保护范围。
本发明提供了一对苯并吡喃对映异构体,该对映异构体具有式1a和式1b的结构:
本发明还提供了所述苯并吡喃对映异构体的制备方法,具体按以下步骤实现:
1)将秦艽药材洗净晾干,粉碎后用70%乙醇浸泡提取3-4次,每次24-48h,将提取液减压浓缩,得浸膏;浸膏加水搅匀,依次用石油醚,乙酸乙酯,正丁醇萃取,浓缩回收正丁醇萃取相得正丁醇相浸膏;
2)将正丁醇相浸膏进行硅胶柱层析,用体积比为80:1,50:1,30:1,15:1及0:1的二氯甲烷/甲醇进行梯度洗脱,收集体积比为15:1的洗脱液,减压浓缩,再通过半制备高效液相色谱,用Agilent Eclipse XDB-C18(9.4×250mm,5μm,3ml/min)分离,得到苯并吡喃类化合物1;
3)将得到的化合物1通过CHIRALPAK IH手性柱进行HPLC手性分离得到化合物1a和化合物1b。
步骤2)中,半制备高效液相色谱分离流动相为体积比为30:70的甲醇/水,进样量为40μL,紫外检测波长为254nm。
步骤3)中,HPLC手性分离采用体积比为50:50的正己烷/异丙醇进行等度洗脱,进样量为20μL,进样速度为1.5ml/min,紫外检测波长为254nm。
本发明还提供了所述苯并吡喃对映异构体或药物组合物在制备保肝药物中的应用。
本发明进一步提供了一种药物组合物,其包括所述苯并吡喃对映异构体或其混合物以及药学上可接受的载体。
实施例1
1、将3kg秦艽药材晾干粉碎后以70%乙醇(g/mL=1/13)浸泡提取3次,每次48h,过滤,合并提取液,减压浓缩至无醇味,干燥后得秦艽浸膏658g。
2、取浸膏用水溶解,然后依次用石油醚,乙酸乙酯,正丁醇萃取,各萃取3次,浓缩回收正丁醇萃取相得正丁醇相浸膏68g。
3、将正丁醇相浸膏以1倍样品量的柱层析硅胶拌样,以10倍样品量的柱层析硅胶(W/W)干法上柱,依次用体积比为80:1→50:1→30:1→15:1→0:1二氯甲烷/甲醇梯度洗脱,薄层色谱收集二氯甲烷/甲醇(15:1)洗脱液。
4、将上述流分通过半制备高效液相色谱仪(甲醇/水=30/70,tR=7.5min,得到苯并吡喃化合物1。参照文献方法,通过CHIRALPAK IH手性柱对其进行HP LC手性分离(正己烷:异丙醇=50:50,进样量=20μL),得到对映异构体(化合物1a,1.8mg,tR=5min,1.5ml/min;化合物1b,1.6mg,tR=6min,1.5ml/min)。
下面对实施例1所得的化合物1、化合物1a及化合物1b进行相关波谱解。
化合物1,白色无定型粉末。平面结构:(+)-HR-ESI-MS m/z 267.0879[M–H]-(C13H16O6H-,计算值267.0874),提示其分子式为C13H16O6,不饱和度为6。
红外光谱(图1)提示有羟基(3371cm-1)、羰基酸(1622cm-1)和甲基(1383cm-1),且两个甲基在同一碳上。13C-NMR谱图共显示13个碳信号,其中包括5个季碳,1个羧基,4个次甲基,2个甲基,1个甲氧基。结合1H-NMR、13C-NMR及HSQC波谱(图2,3,5)提示,在低场处存在2个孤立的芳烃质子7.81(s,1H,H-5),6.37(s,1H,H-8),提示存在3,4苯环-OCH3和-COOH取代;高场区δH 1.44(s,3H,H-9)和δH 1.19(s,3H,H-10)处存在偕二甲基;4.43(d,J=8.3Hz,1H,H-4)和3.49(d,J=8.2Hz,1H,H-3)处存在共振。1H-1H COSY谱图(图4)中,δH 3.49(d,J=8.2Hz,1H,H-3)与4.43(d,J=8.3Hz,1H,H-4)存在相关;δH 1.19(s,3H,H-10)与δH 1.44(s,3H,H-9)存在相关;δH 4.43(d,J=8.3Hz,1H,H-4)与7.81(s,1H,H-5)相关,根据上述谱图信息推测该化合物为2,2-二甲基-2H-苯并吡喃类化合物。
HMBC谱图显示(图6),δH 7.81(s,1H,H-5)、6.37(s,1H,H-8))与δC 176.26(C-11)相关,δH 3.49(d,J=8.2Hz,1H,H-3)、7.81(s,1H,H-5)与δC 69.79(C-4)相关,而1H-1HROESY谱图(图7)中,δH 3.79(s,3H,-OCH3)与6.37(s,1H,H-8)相关,因此,C-6取代基为-COOH,C-7位为-OCH3。由此推导出化合物1的平面结构如示1所示。
绝对构型:化合物1旋光度为推测该化合物可能为外消旋体,即存在两对可能的对映异构体,“3S,4S”,“3R,4R”与“3R,4S”,“3S,4R”。为了确定构型,分别对化合物1a及化合物1b的ECD谱图进行计算拟合(图8)。结果表明,化合物1a的ECD谱与“3R,4R”构型吻合,化合物1b与“3S,4S”构型吻合。因此确定化合物1的两种绝对构型为3R,4R-(1a)和3S,4S-(1b),实验测得其比旋度分别为:
表1实施例1制备的化合物1的一维核磁共振数据
实验例实施例1制备的化合物1,化合物1a及化合物1b体外保肝活性检测
1、L-O2细胞毒检测
取生长状态稳定的L-O2细胞,以每孔100μL(约5×104个细胞),接种于96孔板中,于37℃、5%CO2培养箱中常规培养24h。设置对照组(含10%胎牛血清、1%的青霉素和链霉素的1640完全培养基)和各个浓度的苯并吡喃化合物给药组(5、10、20、40、80μmol/L),培养24h后,每孔加入10μL CCK-8工作液,孵育1.5h,采用酶标仪检测波长450nm处的吸光度(OD)值,检测各化合物对L-O2的细胞毒活性。
结果:如图9所示,本发明制备的苯并吡喃类化合物1,1a及1b无细胞毒作用,其最佳给药浓度范围为20、40、80μmol/L。
2、对APAP诱导L-O2细胞损伤的活性检测
将处于对数生长期的L-O2细胞接种至96孔板中,培养24h。给予不同浓度(20、40、80μmol/L)的化合物1、1a、1b和阳性药(NAC,0.25mmol/L),培养24h后,各组加入APAP(10mmol/L),孵育24h。收集细胞上清液,于每孔加入10μL CCK-8试剂,孵育1.5h后,检测各组吸光度(OD)值,并计算细胞存活率。
结果:如图10所示(NC组为正常组,AP为模型组,NAC阳性组;#p<0.05,##p<0.01,###p<0.001和####p<0.0001与正常组比较;*p<0.05,**p<0.01,
***p<0.001和****p<0.0001与模型组比较),本发明制备的化合物1,1a,1b均具有一定的抗APAP诱导的L-O2细胞损伤活性,且活性优于阳性对照药物N-乙酰半胱胺酸。
应说明的是,以上实施例仅用以说明本发明的技术方案而非限制,尽管参照较佳实施例对本发明进行了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的精神和范围,其均应涵盖在本发明的权利要求范围当中。
Claims (6)
1.一对苯并吡喃对映异构体,其特征在于,该对映异构体具有式1a和式1b的结构:
。
2.权利要求1所述苯并吡喃对映异构体的制备方法,其特征在于,按以下步骤实现:
1)将秦艽药材洗净晾干,粉碎后用70%乙醇浸泡提取3-4次,每次24-48h,将提取液减压浓缩,得浸膏;浸膏加水搅匀,依次用石油醚,乙酸乙酯,正丁醇萃取,浓缩回收正丁醇萃取相得正丁醇相浸膏;
2)将正丁醇相浸膏进行硅胶柱层析,用体积比为80:1,50:1,30:1,15:1及0:1的二氯甲烷/甲醇进行梯度洗脱,收集体积比为15:1的洗脱液,减压浓缩,再通过半制备高效液相色谱分离得到苯并吡喃类化合物1;色谱柱为Agilent Eclipse XDB-C18,规格为9.4×250mm,5μm,流速为3ml/min;
3)将得到的化合物1通过CHIRALPAK IH手性柱进行HPLC手性分离得到化合物1a和化合物1b。
3.根据权利要求2所述苯并吡喃对映异构体的制备方法,其特征在于,步骤2)中,半制备高效液相色谱分离流动相为体积比为30:70的甲醇/水,进样量为40μL,紫外检测波长为254 nm。
4.根据权利要求2所述苯并吡喃对映异构体的制备方法,其特征在于,步骤3)中,HPLC手性分离采用体积比为50:50的正己烷/异丙醇进行等度洗脱,进样量为20μL,进样速度为1.5ml/min,紫外检测波长为254 nm。
5.权利要求1所述苯并吡喃对映异构体在制备保肝药物中的应用。
6.一种药物组合物,包括权利要求1所述苯并吡喃对映异构体或其混合物以及药学上可接受的载体。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311704918.XA CN117756763A (zh) | 2023-12-13 | 2023-12-13 | 一种苯并吡喃对映异构体及其制备方法与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202311704918.XA CN117756763A (zh) | 2023-12-13 | 2023-12-13 | 一种苯并吡喃对映异构体及其制备方法与应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN117756763A true CN117756763A (zh) | 2024-03-26 |
Family
ID=90309819
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202311704918.XA Pending CN117756763A (zh) | 2023-12-13 | 2023-12-13 | 一种苯并吡喃对映异构体及其制备方法与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN117756763A (zh) |
-
2023
- 2023-12-13 CN CN202311704918.XA patent/CN117756763A/zh active Pending
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Hu et al. | Anti-inflammatory octahydroindolizine alkaloid enantiomers from Dendrobium crepidatum | |
CN113912482B (zh) | 愈创木烷型倍半萜类化合物及其制备和应用 | |
CN115991692B (zh) | 菘蓝中螺二烯酮木脂素化合物的制备方法及其应用 | |
CN106008543A (zh) | 一种新的二萜类化合物及其制备方法 | |
CN111848565A (zh) | 单萜基双香豆素类化合物、药物组合物及其制备方法和用途 | |
CN115894414A (zh) | 白英中制备的酰胺木脂素类化合物及其制备方法和应用 | |
CN114933617B (zh) | 二苯甲酮二聚体化合物的制备方法 | |
CN114436802B (zh) | 一种杜松烷倍半萜化合物及其制备方法和应用 | |
Zhao et al. | Chiral separation of sesquineolignans from the stems and leaves of Neoshirakia japonica | |
CN117756763A (zh) | 一种苯并吡喃对映异构体及其制备方法与应用 | |
CN115108935A (zh) | 白英中的生物碱类化合物及其制备方法和应用 | |
CN113968869A (zh) | 愈创木烷倍半萜内酯类化合物Artemvulactone及其制备方法和应用 | |
CN111377933B (zh) | 诸葛菜种子提取的生物碱类化合物及其提取方法与应用 | |
CN111253352B (zh) | 一种从中药斑叶兰中提取分离的化合物及其制备方法和应用 | |
CN103626812A (zh) | 天麻中一种新的巴利森苷类化合物及其用途 | |
Dávid et al. | Jacaranone derivatives with antiproliferative activity from Crepis pulchra and relevance of this group of plant metabolites. Plants 2022; 11: 782 | |
CN110638822A (zh) | 一种促进内皮细胞增殖的合欢皮木脂素苷类化合物及应用 | |
CN113024525B (zh) | 吲哚与喹唑酮类生物碱及其医药用途 | |
KR20160050191A (ko) | 신규 네오리그난 화합물 | |
CN113816857B (zh) | 咖啡酰奎宁酸酯类化合物及其制备方法与应用 | |
CN110540530A (zh) | 一种鸡骨香根提取物及其超临近流体提取方法、应用 | |
CN110713473A (zh) | 降碳新木脂烷类化合物及其医药用途 | |
CN114805382B (zh) | 一种倍半萜色酮类化合物及其分离和在制备抗胰腺癌药物中的应用 | |
CN114957272B (zh) | 一种色原烷二聚体及其制备方法和应用 | |
CN112920146B (zh) | 倍半萜类化合物及其制备方法和在制备抗炎药物中的应用 |
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 |