CN115677520A - Diterpene compound and preparation method and application thereof - Google Patents
Diterpene compound and preparation method and application thereof Download PDFInfo
- Publication number
- CN115677520A CN115677520A CN202211457361.XA CN202211457361A CN115677520A CN 115677520 A CN115677520 A CN 115677520A CN 202211457361 A CN202211457361 A CN 202211457361A CN 115677520 A CN115677520 A CN 115677520A
- Authority
- CN
- China
- Prior art keywords
- fraction
- column chromatography
- ethyl acetate
- fractions
- compound
- 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.)
- Granted
Links
- -1 Diterpene compound Chemical class 0.000 title claims abstract description 24
- 229930004069 diterpene Natural products 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 150000001875 compounds Chemical class 0.000 claims abstract description 32
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 28
- 238000010898 silica gel chromatography Methods 0.000 claims abstract description 18
- 241001164374 Calyx Species 0.000 claims abstract description 13
- 230000002085 persistent effect Effects 0.000 claims abstract description 13
- 239000002023 wood Substances 0.000 claims abstract description 13
- 238000004440 column chromatography Methods 0.000 claims abstract description 10
- 238000000034 method Methods 0.000 claims abstract description 10
- 238000000605 extraction Methods 0.000 claims abstract description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 29
- 238000002156 mixing Methods 0.000 claims description 22
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 20
- 239000003208 petroleum Substances 0.000 claims description 20
- 239000000284 extract Substances 0.000 claims description 14
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 12
- 238000010992 reflux Methods 0.000 claims description 10
- WGLUMOCWFMKWIL-UHFFFAOYSA-N dichloromethane;methanol Chemical compound OC.ClCCl WGLUMOCWFMKWIL-UHFFFAOYSA-N 0.000 claims description 8
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims description 8
- 238000002953 preparative HPLC Methods 0.000 claims description 8
- 150000003839 salts Chemical class 0.000 claims description 8
- 239000003814 drug Substances 0.000 claims description 7
- 238000005520 cutting process Methods 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 6
- 230000002829 reductive effect Effects 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 5
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 claims description 4
- 239000002038 ethyl acetate fraction Substances 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 125000001033 ether group Chemical group 0.000 claims description 2
- 239000000243 solution Substances 0.000 claims 3
- 239000007864 aqueous solution Substances 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 230000004112 neuroprotection Effects 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 12
- 230000006907 apoptotic process Effects 0.000 abstract description 4
- 210000002569 neuron Anatomy 0.000 abstract description 4
- 230000004083 survival effect Effects 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 3
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 3
- 230000005764 inhibitory process Effects 0.000 abstract description 3
- 238000011894 semi-preparative HPLC Methods 0.000 abstract description 2
- 210000004556 brain Anatomy 0.000 abstract 1
- 229940125782 compound 2 Drugs 0.000 description 15
- 229940125904 compound 1 Drugs 0.000 description 12
- 150000001336 alkenes Chemical class 0.000 description 10
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 8
- 238000001228 spectrum Methods 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 6
- 238000005100 correlation spectroscopy Methods 0.000 description 6
- 125000000555 isopropenyl group Chemical group [H]\C([H])=C(\*)C([H])([H])[H] 0.000 description 6
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 6
- 230000000324 neuroprotective effect Effects 0.000 description 6
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 4
- 230000003833 cell viability Effects 0.000 description 4
- 239000013078 crystal Substances 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- QELUYTUMUWHWMC-UHFFFAOYSA-N edaravone Chemical compound O=C1CC(C)=NN1C1=CC=CC=C1 QELUYTUMUWHWMC-UHFFFAOYSA-N 0.000 description 4
- 229950009041 edaravone Drugs 0.000 description 4
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 4
- 238000001819 mass spectrum Methods 0.000 description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 4
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 125000004122 cyclic group Chemical group 0.000 description 3
- 239000002024 ethyl acetate extract Substances 0.000 description 3
- 239000004090 neuroprotective agent Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- 208000019901 Anxiety disease Diseases 0.000 description 2
- 206010008190 Cerebrovascular accident Diseases 0.000 description 2
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 2
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 208000006011 Stroke Diseases 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- ABRVLXLNVJHDRQ-UHFFFAOYSA-N [2-pyridin-3-yl-6-(trifluoromethyl)pyridin-4-yl]methanamine Chemical compound FC(C1=CC(=CC(=N1)C=1C=NC=CC=1)CN)(F)F ABRVLXLNVJHDRQ-UHFFFAOYSA-N 0.000 description 2
- 230000036506 anxiety Effects 0.000 description 2
- 125000002837 carbocyclic group Chemical group 0.000 description 2
- 230000002490 cerebral effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000002983 circular dichroism Methods 0.000 description 2
- 208000010877 cognitive disease Diseases 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012894 fetal calf serum Substances 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 238000005570 heteronuclear single quantum coherence Methods 0.000 description 2
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 125000000468 ketone group Chemical group 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 2
- TZIHFWKZFHZASV-UHFFFAOYSA-N methyl formate Chemical group COC=O TZIHFWKZFHZASV-UHFFFAOYSA-N 0.000 description 2
- 230000004770 neurodegeneration Effects 0.000 description 2
- 208000015122 neurodegenerative disease Diseases 0.000 description 2
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 2
- YNPNZTXNASCQKK-UHFFFAOYSA-N phenanthrene Chemical compound C1=CC=C2C3=CC=CC=C3C=CC2=C1 YNPNZTXNASCQKK-UHFFFAOYSA-N 0.000 description 2
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 2
- 238000000746 purification Methods 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
- 229960005322 streptomycin Drugs 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- VTNULXUEOJMRKZ-UHFFFAOYSA-N 3-[4-(aminomethyl)-6-(trifluoromethyl)pyridin-2-yl]oxy-N-(2H-tetrazol-5-ylmethyl)benzamide Chemical compound N=1NN=NC=1CNC(C1=CC(=CC=C1)OC1=NC(=CC(=C1)CN)C(F)(F)F)=O VTNULXUEOJMRKZ-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
- 208000028698 Cognitive impairment Diseases 0.000 description 1
- 241000221017 Euphorbiaceae Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 208000019695 Migraine disease Diseases 0.000 description 1
- 206010028980 Neoplasm 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
- 206010037660 Pyrexia Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 238000011914 asymmetric synthesis Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000002212 electronic circular dichroism spectrum Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 229930188222 fimbricalyx Natural products 0.000 description 1
- 229930182486 flavonoid glycoside Natural products 0.000 description 1
- 150000007955 flavonoid glycosides Chemical class 0.000 description 1
- 150000004677 hydrates Chemical class 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- OKKJLVBELUTLKV-VMNATFBRSA-N methanol-d1 Chemical compound [2H]OC OKKJLVBELUTLKV-VMNATFBRSA-N 0.000 description 1
- 206010027599 migraine Diseases 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- AICOOMRHRUFYCM-ZRRPKQBOSA-N oxazine, 1 Chemical compound C([C@@H]1[C@H](C(C[C@]2(C)[C@@H]([C@H](C)N(C)C)[C@H](O)C[C@]21C)=O)CC1=CC2)C[C@H]1[C@@]1(C)[C@H]2N=C(C(C)C)OC1 AICOOMRHRUFYCM-ZRRPKQBOSA-N 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 229940002612 prodrug Drugs 0.000 description 1
- 239000000651 prodrug Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 235000007586 terpenes Nutrition 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Landscapes
- Steroid Compounds (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention relates to the technical field of medical biology, in particular to a diterpene compound in persistent calyx wood and a preparation method and application thereof, wherein the diterpene compound is successfully obtained by adopting alcohol extraction, silica gel column chromatography, MCI column chromatography and semi-preparative HPLC (high performance liquid chromatography), the operation method is simple and rapid, the survival rate of PC12 (human brain) cells damaged by a model can be obviously improved, and the compound has a good inhibition effect on PC12 nerve cell apoptosis.
Description
Technical Field
The invention belongs to the technical field of medical biology, and particularly relates to a diterpene compound in persistent calyx wood, a preparation method thereof and application thereof in development of neuroprotective drugs.
Background
The wood of persistent calyx (Strophilobaculia fimbricalyx Borl.) is a perennial shrub plant of the genus Euphorbiaceae. Mainly distributed in the southern Hainan, yunnan, guangxi and Thailand of China. The Thailand folk is used for treating cancer, fever, migraine and the like. However, few reports on the chemical components of the persistent calyx wood are reported at home and abroad. At present, more than thirty compounds such as phenanthrene, terpene and flavonoid glycoside are mainly reported to be separated from the extract, and the material basis is not clear. Therefore, intensive research on chemical components of the persistent calyx wood, separation and preparation of active components thereof and development of novel medicaments are urgently needed.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a Strophafimbrins novel diterpene compound extracted and separated from persistent calyx wood, a preparation method and application thereof, and researches show that the novel skeleton structure compound has a good inhibition effect on PC12 cell apoptosis. Meanwhile, the invention provides a simple, convenient, rapid and high-purity extraction and separation method for the novel compound.
The purpose of the invention is realized by the following technical scheme that the diterpene compound has a general formula shown in a formula (I):
wherein R is 1 is-H or = O; r 2 is-OH, -OCH 3 or-NH 2 ，R 3 is-OH or-OCH 3 。
The stroophilimbriins of the novel 5/6/7 tricyclic system having neuroprotective activity of the present invention and derivatives thereof, and acid addition salts, hydrates or prodrugs of the compounds of the formula (I) of the present invention are also included in the present invention.
Preferably, the diterpene compound is one wherein R 1 Is = O, R 2 is-NH 2 ，R 3 is-OCH 3 The structure ofThe formula is shown as formula 1:
the chemical name of formula 1 is strophilofombrin A.
Preferably, the diterpene compound is a compound represented by formula (I) or (II) 1 is-H, R 2 is-OCH 3 ，R 3 is-OCH 3 The structural formula is shown as formula 2:
the chemical name of formula 2 is strophilofombrin B.
The preparation method of the strophilofombrin A diterpenoid compound comprises the following steps:
a, cutting medicinal materials of the persistent calyx wood into pieces, performing reflux extraction by using ethanol, recovering an extracting solution, and performing reduced pressure concentration and drying to obtain a total extract;
dispersing the total extract by distilled water, and sequentially extracting by using petroleum ether and ethyl acetate with equal volumes to obtain a petroleum ether part and an ethyl acetate part;
c, performing silica gel column chromatography on the ethyl acetate part extract, performing gradient elution by using petroleum ether-ethyl acetate according to the volume ratio of 30: 1, 15: 1, 8: 1, 5: 1, 3: 1, 2: 1, 1: 2, 1: 5 and 0: 1, and mixing to obtain 11 fractions;
subjecting the fraction 11 to silica gel column chromatography, gradient eluting with dichloromethane-methanol at volume ratio of 1: 0, 50: 1, 30: 1, 20: 1, 15: 1, 10: 1, 5: 1, 1: 1, and 0: 1, and mixing to obtain 9 fractions;
subjecting fraction 1 to MCI column chromatography, gradient eluting with methanol-water at volume ratio of 2: 8, 3: 7, 4: 6, 5: 5, 6: 4, 7: 3, 8: 2, 9: 1, and 1: 0, and mixing to obtain 10 fractions;
the fraction 9 was purified by semi-preparative high performance liquid chromatography, eluted with acetonitrile-water at a volume ratio of 41: 59, and the main peak was collected to obtain the compound stropheofombrin A.
Preferably, ethanol in the step a is extracted by refluxing at 85 ℃ for 4 times by using 95% ethanol water solution by volume fraction.
The preparation method of the strophilofombrin B diterpenoid compound comprises the following steps:
step S1: cutting the medicinal material of persistent calyx wood, extracting with ethanol under reflux, recovering extractive solution, concentrating under reduced pressure, and drying to obtain total extract;
step S2: dispersing the total extract with distilled water, and sequentially extracting with petroleum ether and ethyl acetate to obtain petroleum ether fraction and ethyl acetate fraction;
and step S3: subjecting the ethyl acetate extract to silica gel column chromatography, gradient eluting with petroleum ether-ethyl acetate at volume ratio of 30: 1, 15: 1, 8: 1, 5: 1, 3: 1, 2: 1, 1: 2, 1: 5, and 0: 1, and mixing to obtain 11 fractions;
subjecting the fraction 11 to silica gel column chromatography, gradient eluting with dichloromethane-methanol at volume ratio of 1: 0, 50: 1, 30: 1, 20: 1, 15: 1, 10: 1, 5: 1, 1: 1, and 0: 1, and mixing to obtain 9 fractions;
subjecting fraction 1 to MCI column chromatography, gradient eluting with methanol-water at volume ratio of 2: 8, 3: 7, 4: 6, 5: 5, 6: 4, 7: 3, 8: 2, 9: 1, and 1: 0, and mixing to obtain 10 fractions;
purifying the flow 8 by using semi-preparative high performance liquid chromatography, eluting by using methanol-water with the volume ratio of 3: 2, and collecting a main peak to obtain a compound stropheofombrin B.
Preferably, ethanol in the step S3 is extracted by reflux for 4 times at 85 ℃ by adopting ethanol water solution with the volume fraction of 95%.
The diterpene compound or the tautomer, the regioisomer, the stereoisomer, the enantiomer and the diastereoisomer thereof and the structure or the pharmaceutically acceptable salt with the same skeleton or containing the skeleton are applied to the preparation of the neuroprotective medicament.
The term "pharmaceutically acceptable salts" refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, the pharmaceutically acceptable salts described in detail by Berge et al in j. Pharmaceutical Sciences (1977) 66.
The compounds of the present invention may include one or more asymmetric centers, and thus may exist in a variety of stereoisomeric forms, e.g., enantiomeric and/or diastereomeric forms. For example, the compounds of the present invention may be individual enantiomers, diastereomers or geometric isomers (e.g., cis and trans isomers), or may be in the form of mixtures of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers may be separated from mixtures by methods known to those skilled in the art, including: chiral High Pressure Liquid Chromatography (HPLC) and the formation and crystallization of chiral salts; alternatively, preferred isomers may be prepared by asymmetric synthesis.
The invention has the following beneficial effects: (1) The invention provides a new compound Strophahizombriprins diterpenoid compound derived from persistent calyx wood and an extraction, separation and purification method aiming at the new compound, the new compound is successfully obtained by adopting alcohol extraction, silica gel column chromatography, MCI column chromatography and semi-preparative HPLC for separation and purification, the operation method is simple and rapid, and the purity of the compound separated by the method is higher than 95 percent.
(2) Research shows that the novel compound Strophilofofmrins diterpene compound can remarkably improve the survival rate of model injured nerve cells PC12, and has a good inhibition effect on the apoptosis of the nerve cells PC12, so that the compound has remarkable neuroprotective activity, and can be applied to the preparation of novel neuroprotective drugs, such as treatment of but not limited to depression, anxiety and cerebral apoplexy, and improvement of cognitive disorder and motor retardation of patients with neurodegenerative diseases.
Drawings
FIG. 1 is a two-dimensional correlation plot of HMBC for compound 1 and compound 2;
figure 2 is a graph of the structures of compound 1 and compound 2 and their neuroprotective effects on OGD-injured PC12 cells (mean ± SD, n = 4). Compared with the control group, the compound of the formula, ### p<0.001，*p<0.05，**p<0.01，***p<0.001. edaravone (Eda) as a positive control;
FIG. 3 is an experimental and calculated ECD spectrum for compound 1 and compound 2;
FIG. 4 shows the preparation of Compound 1 1 H-NMR spectrum;
FIG. 5 is a drawing of Compound 1 13 C-NMR spectrum;
FIG. 6 is an HMBC spectrum of compound 1;
FIG. 7 is a HSQC spectrum of Compound 1;
FIG. 8 is a drawing of Compound 1 1 H- 1 H COSY spectrogram;
FIG. 9 is a NOESY spectrum of Compound 1
FIG. 10 is a high resolution mass spectrum of Compound 1
FIG. 11 shows single crystal data of Compound 1
FIG. 12 shows preparation of Compound 2 1 H-NMR spectrum;
FIG. 13 is a drawing of Compound 2 13 A C-NMR spectrum;
FIG. 14 is an HMBC spectrum of compound 2;
FIG. 15 is an HSQC spectrum of Compound 2;
FIG. 16 is a drawing of Compound 2 1 H- 1 H COSY spectrogram;
FIG. 17 is a NOESY spectrum of Compound 2;
FIG. 18 is a high resolution mass spectrum of Compound 2.
Detailed Description
The following examples are provided to further illustrate the present invention and should not be construed as limiting the scope thereof.
A diterpene compound having the general formula shown in formula (I):
wherein R is 1 is-H or = O; r 2 is-OH, -OCH 3 or-NH 2 ，R 3 is-OH or-OCH 3 。
Example 1: preparation of Compound 1 (Strophilofofmrin A)
Cutting and crushing 18kg of medicinal plant of the persistent calyx wood, extracting for 4 times by refluxing at 85 ℃ with 95% ethanol, combining the extracting solutions, and concentrating and drying under reduced pressure to obtain a total extract (472 g). Dispersing the total extract with 10 times of distilled water, and sequentially extracting with petroleum ether and ethyl acetate with equal volume to obtain petroleum ether fraction (123 g) and ethyl acetate fraction (55 g). Subjecting the ethyl acetate extract to silica gel column chromatography, gradient eluting with petroleum ether-ethyl acetate at volume ratio of 30: 1, 15: 1, 8: 1, 5: 1, 3: 1, 2: 1, 1: 2, 1: 5, and 0: 1, and mixing to obtain 11 fractions; subjecting the fraction 11 to silica gel column chromatography, gradient eluting with dichloromethane-methanol at volume ratio of 1: 0, 50: 1, 30: 1, 20: 1, 15: 1, 10: 1, 5: 1, 1: 1, and 0: 1, and mixing to obtain 9 fractions; subjecting fraction 1 to MCI column chromatography, gradient eluting with methanol-water at volume ratio of 2: 8, 3: 7, 4: 6, 5: 5, 6: 4, 7: 3, 8: 2, 9: 1, and 1: 0, and mixing to obtain 10 fractions; purifying the fraction 9 by semi-preparative high performance liquid chromatography, eluting with acetonitrile-water at a volume ratio of 41: 59, and collecting the main peak to obtain the compound 1.
Example 2: preparation of Compound 2 (Strophilofofmrin B)
Cutting and crushing 18kg of medicinal plant of the persistent calyx wood, extracting for 4 times by refluxing at 85 ℃ with 95% ethanol, combining the extracting solutions, and concentrating and drying under reduced pressure to obtain a total extract (472 g). Dispersing the total extract with 10 times of distilled water, and sequentially extracting with petroleum ether and ethyl acetate with equal volume to obtain petroleum ether fraction (123 g) and ethyl acetate fraction (55 g). Subjecting the ethyl acetate extract to silica gel column chromatography, gradient eluting with petroleum ether-ethyl acetate at volume ratio of 30: 1, 15: 1, 8: 1, 5: 1, 3: 1, 2: 1, 1: 2, 1: 5, and 0: 1, and mixing to obtain 11 fractions; subjecting the fraction 11 to silica gel column chromatography, gradient eluting with dichloromethane-methanol at volume ratio of 1: 0, 50: 1, 30: 1, 20: 1, 15: 1, 10: 1, 5: 1, 1: 1, and 0: 1, and mixing to obtain 9 fractions; subjecting fraction 1 to MCI column chromatography, gradient eluting with methanol-water at volume ratio of 2: 8, 3: 7, 4: 6, 5: 5, 6: 4, 7: 3, 8: 2, 9: 1, and 1: 0, and mixing to obtain 10 fractions; purifying the fraction 8 by semi-preparative high performance liquid chromatography, eluting with methanol-water at volume ratio of 3: 2, and collecting main peak to obtain the compound 2.
The structural formulas of compound 1 and compound 2 are as follows:
and (3) structure analysis process:
compound 1: the novel compound prepared in example 1 of the present invention is yellow needle crystal. The high resolution mass spectrum gives the excimer ion peak m/z 350.1346[ M ] +Na ]]+ (calculated 350.1363), combined 1 H-NMR and 13 C-NMR (Table 1) confirmed that the compound has the formula C 19 H 21 NO 4 The unsaturation was calculated to be 10. 1 H、 13 C-NMR and HSQC show three methyl groups (two methoxy groups), three methylene groups (one of which is an alkene), four methine groups (two of which are alkenes), and nine quaternary carbons (three carbonyl groups and five of which are alkenes). From Me-18 (. Delta.) according to the analysis H 1.87 To C-4 (. Delta.) C 143.5)、C-19(δ C 114.2 And C-5 (. Delta.) C 49.4 From the terminal alkene methylene H-19 to Me-18, C-4 (. Delta.) C 147.1)、C-19(δ C 114.2 And C-5 (. Delta.) C 52.3 And from OMe-3 (. Delta.) H 3.81,3H,s) to C-3 (. Delta.) C 177.4 HMBC correlation data (FIG. 1) can confirm the presence of typical isopropenyl groups (. Delta.) H 4.89,5.07，2H，s；δ C 114.8,147.1；δ H 1.87，3H，s；δ C 24.1 And methyl ester carbonyl (. Delta.) and C 175.6,52.3). Six remaining olefin quaternary carbons (. Delta.) C 197.4, 172.1, 160.9, 158.5,136.5 and 137.9) and two olefinic methines form three further pairs of double bonds and a conjugated carbonyl group (. Delta.) ( C 177.5). The above functional groups account for 7 of 10 unsaturations. Thus, remainingThe 3 unsaturation requirement is a tricyclic system in the molecule.
In the HMBC spectra (FIG. 1), the correlation originates from Me-20 (δ) H 1.19 To C-1, C-9, C-10, me-20, from Me-18 and H-19 to C-5, typically isopropenyl, from H-2 (. Delta.) (delta.) H 4.25 To C-1, C-11 and C-3, and continuous correlation 1 H- 1 H-5/H-6 α/H-7 α/H-7 β in the H COSY spectrum establishes a fragment C-5 of the six-membered carbocyclic ring B, which is part of the tricyclic structure, substituted by a methyl group at C-10 and typically an isopropenyl group. 1 H- 1 The H COSY spectrum also shows the presence of a spin system H-2/H-1 α/H-1 β, and also from H-2 (δ) H 4.25 HMBC correlation of 3 methyl formate groups to C-1, C-11 and C-3, from Me-20 to C-1 and C-9, from H-1 (. Delta.) H 2.28, 2.14) to C-20, C-9, C-11 and C-3, a five-membered ring A substituted at C-2 with a methyl ester carbonyl group is constructed which is bonded to ring B at common C-9 and C-10. Then, by the reaction of alpha, beta unsaturated ketone groups (. Delta.) C 160.9，C-9；δ C 137.9，C-11；δ C 158.5, C-17) from two olefinic methines H-13 (. Delta.) H 6.89 To C-8, C-12, and H-14 (. Delta.) H 7.23 To C-9 and C-17, and a spin system H-13/H-14, constructed seven-membered ring C. The HMBC cross peaks of H-14/C-7, H-7/C-14, and H-2/C-17 further indicate that Ring C shares C-8 and C-9 with Ring B and C-9 and C-11 with Ring A. Thus, the planar structure of strophilofibrin A with the novel 5/6/7 ring system is elucidated. In addition, the molecular structure of the compound has chirality, and the configuration of the compound is determined to be (2R, 5R, 10R) by calculating circular dichroism (figure 2). The compound was also single crystal tested and the crystal structure is shown in the figure (fig. 11), and in conclusion, the structure of the compound was determined and named strophilofibrin a.
Compound 2: high resolution mass spectrum gives excimer ion peak m/z 351.1572[ M + Na ]]+ (calculated 351.1567), combined 1 H-NMR and 13 C-NMR (Table 1) to confirm that the compound has the formula C 20 H 24 O 4 The unsaturation was calculated to be 9. 1 H、 13 C-NMR and HSQC show four methyl groups (two methoxy groups), four methylene groups (one of which is an alkene),Four methines (two of which are alkenes) and eight quaternary carbons (two carbonyls and five of which are alkenes). From Me-18 (. Delta.) according to the analysis H 1.87 To C-4 (. Delta.) C 147.1)、C-19(δ C 114.2 And C-5 (. Delta.) C 51.8 From the terminal alkene methylene H-19 to Me-18, C-4 (. Delta.) C 147.1)、C-19(δ C 114.2 And C-5 (. Delta.) C 51.8 And from OMe-3 (. Delta.) H 3.70,3H,s) to C-3 (. Delta.), (S) C 177.4 HMBC correlation data, it can be confirmed that a typical isopropenyl group (. Delta.) exists H 4.85,5.00，2H，s；δ C 114.2,147.1；δ H 1.87，3H，s；δ C 24.2 And methyl ester carbonyl (. Delta.) and C 177.4,52.5). The remaining five olefin quaternary carbons (. Delta.) C 177.5, 164.3, 160.8, 146.5 and 138.4) and two olefinic methines (. Delta.) ( H 7.05 and 7.23,2h, d, J =10.2 Hz) constitute three further pairs of double bonds and one conjugated carbonyl group (δ) C 177.5). The functional groups account for 6 of the 9 unsaturations. Thus, the remaining 3 unsaturations requirement is a tricyclic ring system in the molecule.
In the HMBC spectra (FIG. 1), the correlation originates from Me-20 (δ) H 1.11 From C-5, C-9, from typically isopropenyl Me-18 and H-19 to C-5, from H-5 (. Delta.) H 2.44 To C-4, C-10 and C-20, from H-7 (. Delta.) H 3.11, 2.92) to C-5, C-6, C-8 and C-9, and continuous correlation 1 H- 1 H-5/H-6 α/H-7 α/H-7 β in the H COSY spectrum establishes a fragment C-5 of the six-membered carbocyclic ring B, which is part of the tricyclic structure, substituted by a methyl group at C-10 and typically an isopropenyl group. 1 H- 1 The H COSY spectrum also shows the presence of a spin system H-2/H-1 α/H-1 β, and from H-2 (δ) H 4.15 HMBC correlation of 3 methyl formate groups to C-1, C-11 and C-, from Me-20 to C-1 and C-9, from H-1 (. Delta.) H 2.25, 2.01) to C-20, C-9, C-11 and C-3, a five-membered ring A substituted at C-2 with a methyl ester carbonyl group is constructed which is bonded to ring B at common C-9 and C-10. Then, by the reaction of alpha, beta unsaturated ketone groups (. Delta.) C 160.8，C-9；δ C 146.5，C-11；δ C 177.5, C-17 or delta C 115.1，C-13；δ C 164.3,C-12；δ C 177.5, C-17) and from other OMe-12 toC-12(δ C 164.3 From two olefinic methines H-13 (. Delta.) H 7.05 To C-8, C-12 and C-17, and H-14 (. Delta.) H 7.23 Analysis of the HMBC correlations to C-9 and C-12, and a spin system H-13/H-14, seven-membered ring C was constructed. The HMBC cross peaks of H-14/C-7, H-7/C-14, and H-2/C-17 further indicate that Ring C shares C-8 and C-9 with Ring B and C-9 and C-11 with Ring A. Thus, a planar structure with a novel 5/6/7 ring system is illustrated. We additionally determined that the configuration was likewise (2R, 5R, 10R) by computational circular dichroism (FIG. 2). In conclusion, the structure of the compound is determined and named Strophiofimbrin B.
Table 1: of Compound 1 and Compound 2 1 H NMR and 13 c NMR data
a Recorded in CDCl 3 ； b Recorded in MeOD.
Active part:
experimental materials: cell culture solution DMEM, an air-permeable cell culture bottle, a 96-well cell culture plate, PC12 cells, fetal bovine serum, trypsin, a CCK8 kit, a penicillin-streptomycin double antibody solution (penicillin working concentration is 100U/mL, streptomycin working concentration is 0.1 mg/L), strophilobimbrin A, strophilobimbrin B, edaravone, phosphate buffer PBS (0.03 mol/L, pH = 7.2), a microporous filter, a syringe, ethanol, DMSO, a centrifuge tube, an EP tube, a 1mL tip, a 200ul tip, a glove, a mask and the like.
Purchased PC12 cells at 37 ℃ 5% 2 And (5) performing adaptive culture in an incubator. The culture solution is DMEM (containing 5% fetal calf serum and 1% double antibody). At 37 ℃ 5% CO 2 Culturing in a saturated humidity incubator, pouring out the culture solution after the cells are full of the bottom of the culture flask, washing with PBS for 2 times, adding trypsin for digestion for 2min, adding DMEM culture solution to stop digestion, gently blowing, centrifuging at 1200r/min for 5min, re-suspending the cell precipitate, inoculating to the culture flask, observing and photographing under a microscope, and taking a picture when the cells are full of a monolayer, namelyCan be used for experiment.
Establishing an experimental model, grouping, modeling and dosing:
establishing Oxygen sugar Deprivation-reoxygenation (OGD/R) model, and diluting the treated cells to 5 × 10 with DMEM culture solution 4 one/mL cell suspension was inoculated into 96-well plates, 200. Mu.L per well and cultured for 24h. The experiment was divided into a normal control group (control group), a model group (OGD group), an experimental group (stropheofombrin a, stropheofombrin B drug group), and a positive drug control group (edaravone group): changing the total amount of the normal control group into DMEM culture solution (containing 10% fetal calf serum), and culturing in a constant-temperature incubator; the total amount of the model group, experimental group and positive drug control group is changed into DMEM sugar-free culture solution at a content of 1% 2 、94％N 2 And 5% of CO 2 The three air culture boxes are subjected to low oxygen treatment for 6 hours, samples with final concentrations of 6.25 mu M, 12.5 mu M, 25 mu M, 50 mu M and 100 mu M are respectively added into each hole of an experimental group, edatavone with corresponding concentrations is added into a positive control group, and then normal oxygen and reoxygenation are recovered for 24 hours. 10 μ L of CCK8 was added to each well, incubated at 37 ℃ for 2h, and the absorbance (OD) of each group of cells at 450nm was measured with a microplate reader.
The calculation formula is as follows:
cell survival = [ (OD administration group-OD blank)/(OD control group-OD blank) ] × 100%.
The protective effect was evaluated by cell viability, and compared to OGD control (DMSO, cell viability 50.82 ± 1.99%), compounds 1 (50 μ M) and 2 (50 μ M) showed significant neuroprotective effects, cell viability 80.23 ± 3.14% and 81.25 ± 3.83%, respectively, and cell viability of the positive control drug edaravone (50 μ M) 85.95 ± 1.07% (fig. 2). Experimental results show that the compounds 1-2 have a neuroprotective effect on PC12 cells damaged by OGD.
The novel compound can obviously improve the survival rate of the PC12 cells damaged by the model, has a better inhibiting effect on the apoptosis of the PC12 nerve cells, and can be applied to the preparation of novel neuroprotective drugs, such as treatment of but not limited to depression, anxiety and cerebral apoplexy, and improvement of cognitive impairment and motor retardation of patients suffering from neurodegenerative diseases. While the invention has been described in connection with specific embodiments thereof, it is not intended that such description be construed as limiting the scope of the invention, which is defined by the appended claims, as any modification thereto will fall within the scope of the invention.
Claims (10)
1. A diterpene compound has a general formula shown in formula (I):
wherein R is 1 is-H or = O; r 2 is-OH, -OCH 3 or-NH 2 ，R 3 is-OH or-OCH 3 。
2. The diterpene compound according to claim 1, wherein R 1 Is = O, R 2 is-NH 2 ，R 3 is-OCH 3 The structural formula is shown as formula 1:
the chemical name of formula 1 is strophilofombrin A.
3. The diterpene compound according to claim 1, wherein R 1 is-H, R 2 is-OCH 3 ，R 3 is-OCH 3 The structural formula is shown as formula 2:
the chemical name of formula 2 is strophilofombrin B.
4. A process for the preparation of a diterpene compound according to claim 2, which comprises the steps of:
a, cutting the medicinal plant of the persistent calyx wood into pieces, performing reflux extraction by using ethanol, recovering an extracting solution, and performing reduced pressure concentration and drying to obtain a total extract;
dispersing the total extract by distilled water, and sequentially extracting by using petroleum ether and ethyl acetate with equal volumes to obtain a petroleum ether part and an ethyl acetate part;
c, performing silica gel column chromatography on the ethyl acetate part, performing gradient elution by using petroleum ether-ethyl acetate, and combining to obtain 11 fractions; performing silica gel column chromatography on the fraction 11, performing gradient elution by using dichloromethane-methanol, and combining to obtain 9 fractions; subjecting fraction 1 to MCI column chromatography, gradient eluting with methanol-water, and mixing to obtain 10 fractions; the fraction 9 was purified by semi-preparative high performance liquid chromatography, eluted with acetonitrile-water, and the main peak was collected to obtain the compound strophilofombrin A.
5. The process for producing a diterpene compound according to claim 4, wherein the step c comprises the following steps:
subjecting ethyl acetate to silica gel column chromatography, gradient eluting with petroleum ether-ethyl acetate at volume ratio of 30: 1, 15: 1, 8: 1, 5: 1, 3: 1, 2: 1, 1: 2, 1: 5, and 0: 1, and mixing to obtain 11 fractions;
subjecting the fraction 11 to silica gel column chromatography, gradient eluting with dichloromethane-methanol at volume ratio of 1: 0, 50: 1, 30: 1, 20: 1, 15: 1, 10: 1, 5: 1, 1: 1, and 0: 1, and mixing to obtain 9 fractions;
subjecting fraction 1 to MCI column chromatography, gradient eluting with methanol-water at volume ratio of 2: 8, 3: 7, 4: 6, 5: 5, 6: 4, 7: 3, 8: 2, 9: 1, and 1: 0, and mixing to obtain 10 fractions;
the fraction 9 was purified by semi-preparative high performance liquid chromatography, eluted with acetonitrile-water at a volume ratio of 41: 59, and the main peak was collected to obtain the compound stropheofombrin A.
6. The method for producing diterpene compounds according to claim 4, wherein the ethanol in step a is extracted with 95% by volume of an aqueous ethanol solution at 85 ℃ under reflux for 4 times.
7. A process for the preparation of a diterpene compound according to claim 3, which comprises the steps of:
step S1: cutting the medicinal material of persistent calyx wood, extracting with ethanol under reflux, recovering extractive solution, concentrating under reduced pressure, and drying to obtain total extract;
step S2: dispersing the total extract with distilled water, and sequentially extracting with petroleum ether and ethyl acetate to obtain petroleum ether fraction and ethyl acetate fraction;
and step S3: performing silica gel column chromatography on the ethyl acetate part, performing gradient elution by using petroleum ether-ethyl acetate, and mixing to obtain 11 fractions; performing silica gel column chromatography on the fraction 11, performing gradient elution by using dichloromethane-methanol, and mixing to obtain 9 fractions; subjecting the fraction 1 to MCI column chromatography, performing gradient elution with methanol-water, and mixing to obtain 10 fractions; and purifying the flow part 8 by using semi-preparative high performance liquid chromatography, eluting by using methanol-water, and collecting a main peak to obtain the compound strophilofibrin B.
8. The method for preparing diterpene compounds according to claim 7, wherein the step S3 comprises the following steps:
subjecting ethyl acetate to silica gel column chromatography, gradient eluting with petroleum ether-ethyl acetate at volume ratio of 30: 1, 15: 1, 8: 1, 5: 1, 3: 1, 2: 1, 1: 2, 1: 5, and 0: 1, and mixing to obtain 11 fractions;
subjecting the fraction 11 to silica gel column chromatography, gradient eluting with dichloromethane-methanol at volume ratio of 1: 0, 50: 1, 30: 1, 20: 1, 15: 1, 10: 1, 5: 1, 1: 1, and 0: 1, and mixing to obtain 9 fractions;
subjecting fraction 1 to MCI column chromatography, gradient eluting with methanol-water at volume ratio of 2: 8, 3: 7, 4: 6, 5: 5, 6: 4, 7: 3, 8: 2, 9: 1, and 1: 0, and mixing to obtain 10 fractions;
purifying the flow portion 8 by using semi-preparative high performance liquid chromatography, eluting by using methanol-water with the volume ratio of 3: 2, and collecting a main peak to obtain a compound strophilofibrin B.
9. The preparation method according to claim 7, wherein the ethanol in the step S3 is extracted by refluxing at 85 ℃ for 4 times by using an ethanol aqueous solution with a volume fraction of 95%.
10. Use of the diterpene compound according to any one of claims 1 to 3 or a tautomer, regioisomer, stereoisomer, enantiomer, diastereomer or a structurally or pharmaceutically acceptable salt thereof having the same or comprising the same skeleton for the preparation of a medicament for neuroprotection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211457361.XA CN115677520B (en) | 2022-11-18 | 2022-11-18 | Diterpene compound and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211457361.XA CN115677520B (en) | 2022-11-18 | 2022-11-18 | Diterpene compound and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115677520A true CN115677520A (en) | 2023-02-03 |
| CN115677520B CN115677520B (en) | 2024-04-02 |
Family
ID=85053384
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211457361.XA Active CN115677520B (en) | 2022-11-18 | 2022-11-18 | Diterpene compound and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115677520B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101360506A (en) * | 2005-11-25 | 2009-02-04 | 派普林研究股份有限公司 | Methods for wound healing |
| CN105152921A (en) * | 2015-10-22 | 2015-12-16 | 云南民族大学 | Tricyclic diterpenoid compound and preparation method and application thereof |
| CN113683507A (en) * | 2021-08-24 | 2021-11-23 | 沈阳药科大学 | Guaiane sesquiterpene derivative in regachia and application thereof |
| CN114031579A (en) * | 2021-11-11 | 2022-02-11 | 沈阳药科大学 | Preparation and application of daphnane diterpenoid compounds in lilac daphne flower buds |
| CN114874098A (en) * | 2022-06-15 | 2022-08-09 | 扬州大学 | Compound extracted and separated from persistent calyx wood as well as preparation method and application thereof |
-
2022
- 2022-11-18 CN CN202211457361.XA patent/CN115677520B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101360506A (en) * | 2005-11-25 | 2009-02-04 | 派普林研究股份有限公司 | Methods for wound healing |
| CN105152921A (en) * | 2015-10-22 | 2015-12-16 | 云南民族大学 | Tricyclic diterpenoid compound and preparation method and application thereof |
| CN113683507A (en) * | 2021-08-24 | 2021-11-23 | 沈阳药科大学 | Guaiane sesquiterpene derivative in regachia and application thereof |
| CN114031579A (en) * | 2021-11-11 | 2022-02-11 | 沈阳药科大学 | Preparation and application of daphnane diterpenoid compounds in lilac daphne flower buds |
| CN114874098A (en) * | 2022-06-15 | 2022-08-09 | 扬州大学 | Compound extracted and separated from persistent calyx wood as well as preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115677520B (en) | 2024-04-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110452249B (en) | Novel germacrane type sesquiterpene lactone compound and preparation and application thereof | |
| CN113105388B (en) | Euphorbia lathyris diterpene alkyl compound and extraction method and application thereof | |
| TWI648257B (en) | Compounds from antrodia camphorata, method for preparing the same and use thereof | |
| JP2024505106A (en) | Azulene compounds and their preparation methods and uses | |
| WO2017220041A2 (en) | Bezafibrate pharmaceutical composition and application thereof in rheumatoid arthritis | |
| CN108689851B (en) | Tiglic alkane type diterpene compound and preparation method and application thereof | |
| CN105294665A (en) | Novel diterpene compound for neuroprotection | |
| CN110305014B (en) | Cycloneolignanoid enantiomers, preparation and application thereof | |
| CN113912482A (en) | Guaiane type sesquiterpenoids, preparation and application thereof | |
| CN115677520B (en) | Diterpene compound and preparation method and application thereof | |
| CN111205347A (en) | Oleanane-type triterpenoid saponin compound and extraction method and application thereof | |
| CN114874098B (en) | Compound extracted and separated from glaucocalyxa, and preparation method and application thereof | |
| CN105503996A (en) | Limonin compound as well as preparation method and neuroprotective effect thereof | |
| CN109180632B (en) | A method for preparing compound separated from radix Tripterygii Wilfordii | |
| WO2017215679A2 (en) | Pharmaceutical composition of chlorambucil and medical use thereof to counter depression | |
| CN110204589B (en) | Effective component of feather cockscomb seed, extraction method and application thereof in preparing neuroprotective medicament | |
| CN108948040B (en) | Gilmaxane type sesquiterpene compound extracted from herba Centellae and application thereof | |
| CN109206392B (en) | Coumarin compound and preparation method and application thereof | |
| CN106588948B (en) | Oxygen-containing bridge iridoid and its preparation method and application | |
| CN110585221A (en) | Albizzia julibrissin new lignan compound for improving steatosis and application thereof | |
| CN111217886B (en) | Ursolic triterpenoid saponin compound and preparation method and application thereof | |
| CN114957272B (en) | Chromane dimer and preparation method and application thereof | |
| CN113214211B (en) | Compound extracted from peony bark charcoal and preparation method and pharmaceutical application thereof | |
| CN115991692B (en) | Preparation method and application of spirodienone lignan compound in Isatis tinctoria | |
| CN115806491B (en) | Terpenoid with nerve cell protection in corn silk and preparation method and application thereof |
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 |