CN1081920C - Herba pteridis semipinnatae extract and its medicinal composite - Google Patents
Herba pteridis semipinnatae extract and its medicinal composite Download PDFInfo
- Publication number
- CN1081920C CN1081920C CN95119380A CN95119380A CN1081920C CN 1081920 C CN1081920 C CN 1081920C CN 95119380 A CN95119380 A CN 95119380A CN 95119380 A CN95119380 A CN 95119380A CN 1081920 C CN1081920 C CN 1081920C
- Authority
- CN
- China
- Prior art keywords
- cell
- herba pteridis
- extract
- meoh
- 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.)
- Expired - Fee Related
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention discloses two kinds of compounds extracted from pteridaceae plant Herba Pteridis semipinnatae and a medicinal composite of the compounds. The present invention has the technical scheme that the Herba Pteridis semipinnatae and the extract F-A[corresponding-7 alpha, 11 alpha-11 alpha-dihydroxyl-15 oxygen-16 olefin-copy benefit acid lactone (19, 6)] and F-B[corresponding-11 alpha-hydroxy group-15 oxygen-16 olefin-19 copy benefit acid]have very obvious effect on inhibiting tumour and other medicial effects. Indicated by a research, the two kinds of compounds extracted from the Herba Pteridis semipinnatae are novel compounds having medical effect.
Description
The present invention relates to one group of chemical compound that extracts from Pteridaceae plant Herba Pteridis Semipinnatae (Pteris Semipi-nnate Linn), with and medical composition, particularly relate to new antitumor drug.
Malignant tumor is the most serious disease of harm humans health, Chinese scholars antagonism cancerization is learned drug research and is lasted more than 40 year, carried out very extensive studies, about 500,000 kinds of material through screening, what have the certain significance clinically has kind more than 60 approximately, existing anticancer chemotherapy medicine has been obtained bigger progress at anti-curing oncoma, but these based chemotherapy medicine great majority are when killing cancerous cell, to normal histoorgan, as bone marrow, gastrointestinal tract etc. detrimental effect is arranged also, present certain toxicity, thereby the restriction consumption hinders the curative effect performance.This seminar has proved a kind of plant---the extract of Pteridaceae plant Herba Pteridis Semipinnatae has significant antitumaous effect, it uses the proliferation function that remarkable inhibition tumor cell is arranged separately, share with other chemotherapeutic and can promote the chemotherapeutics active anticancer, but can resist the untoward reaction that it suppresses bone marrow and infringement gastrointestinal function, has high-efficiency low-toxicity, the advantage that untoward reaction is few.Yet, what this anticancer main active ingredient of Herba Pteridis Semipinnatae is, in its extract that a kind of be topmost active ingredient, how is the anticancer mechanism of these active ingredients? these active ingredients are a kind of anticarcinogen of high-efficiency low-toxicity new, that be not familiar with by people as yet, and this is a research in the past an open question still.
The objective of the invention is to from Herba Pteridis Semipinnatae, extract a kind of or several have the chemical compound of active anticancer, determine its chemical constitution, understand its anticancer mechanism, for clinical medicine provides a kind of new medical composition, particularly new antitumor drug.
Another object of the present invention is to provide a kind of or several have the chemical compound of medical function, or the medical composition of forming by these chemical compounds.
The present invention relates to following two kinds of chemical compounds, these two kinds of chemical compounds are to extract from Pteridaceae plant Herba Pteridis Semipinnatae at first, separate and confirm through identifying.
1, compound F 17-hydroxy-corticosterone-ACompound F-A:mp243-246 ℃, [α]
D-74 ° (c=0.2, MeOH)
Ent-7 α, 11 α-dihydroxy-15-oxo-Kaur-16-en-19,6 beta-ol ide mapping-7 α, 11 alpha-dihydroxy-s-15 oxygen-16 alkene-copy sharp acid lactone (19,6)
2, compound F 17-hydroxy-corticosterone-BCompound F-B:mp235-239 ℃, [α]
D-145 ° (c=0.2, MeOH)
Ent-11 α-hydroty-15-oxo-Kaur-16en-19-oic acid mapping-11 Alpha-hydroxy-15 oxygen-16 alkene-19 is copied sharp acid
More than the extraction process of two kinds of chemical compounds be:
It is clean air-dry to get the Herba Pteridis Semipinnatae herb, and ethanol extraction 3 times concentrates the back and goes up activated-charcoal column, with MeOH: CHCl
3(2: 1) eluting concentrates the back and goes up the silicagel column separation, with CHCl
3: MeOH (90: 2) eluting gets a series of chemical compounds.Wherein above-mentioned two kinds of chemical compounds are after the experimentation proof has significant pharmacologically active, determined their chemical constitution more through the research of this group, the evaluation of these chemical constitutions Nobutoshi professor Tanaka of Tokyo department of pharmacy of university of science that got back is afterwards helped, do further definite through nuclear magnetic resonance, NMR (NMR) spectrographic technique of etc.ing.
Through two kinds of chemical compounds of said method extraction separation, through human leukemia cell line HL-60 and the erythroleukemia cell strain K of zoopery proof to cultivating
562, nasopharyngeal carcinoma cell (CNE-22), oral squamous carcinoma cell (KB Cell) have tangible active anticancer, further these two kinds of compound anti-cancering activities of the preliminary proof of research may be relevant with the mitosis that suppresses cell.
Above-mentioned two kinds of chemical compounds also can extract from other plant of Pteridaceae and separate, as Pteridaceae plant Herba Pteridis disparis (Pteris dispar L.), and Pteris altissima Poir:Pteristremula R.Br; Pteris livida mett; Also can extract above-mentioned chemical constituent in the plants such as Pteris Longipes Don.Wherein one or more compositions can be extracted or isolate to other plant also.
Above-mentioned two kinds of chemical compounds still can carry out structure of modification or obtain with artificial method is synthetic by similar compound.
Above-mentioned two kinds of chemical compounds are as medical composition, and antineoplastic pharmacologically active particularly is by following experiment confirm.
Embodiment one:
1, materials and methods
The above-claimed cpd F-A of employing through identifying is dissolved among the 5%DMSO, and the reuse culture fluid is diluted to experimental concentration.
Reagent and instrument:
3H-TdR is a Shanghai nuclear technology development company product, be 22ci/mmol than living, the RNA enzyme is a Shanghai Bai Aoke skill company product, nucleic acid mixes agent propidium iodide (PI) dyestuff, NBT (p-nitro-blue-tetrazolium chloride), TPA (12-O-tetradecanoyl-phorbol-13-acetate) is the sigma product, and RPMI-1640 is the Gibco product, the EPICS XL of U.S. COULTER company type flow cytometer.
Tumor cell and cultivation human promyelocytic leukemia HL-60 and people's red white corpuscle leukemia K
562Introduce by institute of Materia Medica,Chinese Academy of Medical Sciences, all containing 10% deactivation calf serum, 100 unit/ml
-1Penicillin and 100 μ gml
-1In the RPMI-1640 culture medium of streptomycin, put the cultivation of going down to posterity in 37 ℃ 5% the CO2 incubator.
Cell inhibitory effect effect and IC
50Measure: the trophophase cell of taking the logarithm is mixed with 1 * 10
5Individual ml
-1Cell suspension.Be inoculated in respectively in the culture plate of 1ml, medicine is divided into 5 variable concentrations, and every concentration is established three parallel pipes, contrast adds commensurability culture fluid, behind medicine and the cells contacting 24hr, expects blue repelling attack living cell counting with platform, calculate inhibitory rate of cell growth, calculate IC by improvement Karber formula again
50
The mensuration of HL-60 cell growth curve, the trophophase cell of taking the logarithm is made into 5 * 10
4Individual ml
-1Cell suspension, in the packing culture plate, every group every days three hole, add medicine then, matched group adds the equivalent culture fluid, puts 37 ℃ of 5%CO
2Cultivate.Take out cell in different time, expect blue repelling attack living cell counting, observed continuously 5 days by platform.
3The H-deoxyribosylthymine (
3H-TdR) mix experiment: the trophophase cell of taking the logarithm is made in certain cell number inoculated and cultured plate, and after every group of four parallel holes, adding medicine were cultivated drug effect different time inner cell counting, every hole added
3It is 1uciml that H-TdR makes final concentration
-1Behind the 4hr, on 49 type all-glass papers, after the processing, in liquid flashing counting determining per minute umber of pulse (CPM), deduction background CPM is with CPM/5 * 10 with bull cell harvestor collecting cell
5Cell is a unit of account.Try to achieve suppression ratio (IR%) by following formula
Influence to HL-60 cell NBT reducing power
[], with cultivating administration group and the cellular control unit of 4d, at 4 ℃ of 1000rmin
-1Centrifugal 5min abandons supernatant, and every pipe adds 0.5ml and contains 1%NBT and 200ngml
-1The culture fluid of TPA, 37 ℃ of insulation 60min, the centrifugal supernatant of abandoning is got cell smear, the wright-Giemsa 10min that dye, oily mirror is observed down, and the black-and-blue first precipitation person of appearance is positive in the cell.200 cells of random observation calculate NBT cell positive rate.
Cell cycle analysis matched group and dosing group cell, centrifugal cell harvesting reaches 1 * 10 at the appointed time
6Individual, add cold PBS liquid washed twice, fix 4 ℃ of preservations with 70% cold ethanol.The centrifugal ethanol of abandoning during mensuration, the PBS washed twice.Add 37 ℃ of RNA enzymes (0.1mg/ pipe) digestion 30 minutes, the PI (50ugml that dyes
-1) 15 minutes, put EPICSXL type flow cytometer (FCM) analysis of cells period profile.
Matched group and administration group cell are got in the preparation of HL-60 cell electron microscope specimen at the appointed time, with cold PBS liquid washing secondary, fully be fixed in 25% or the dialdehyde sodium cacodylate buffer liquid of PH7.2 after the piping and druming, gradient alcohol dehydration, the Epon812 embedding, the section of LKB-V ultramicrotome, the plumbous two dyeing of uranium, H-300 transmissioning electric mirror checking.
2, experimental result
F-A is to HL-60 and K
562Cell inhibiting effect and IC
50Value F-A variable concentrations is to HL-60 and K
562Suppression ratio behind the cytosis 24hr sees Table 1, wherein to the IC of two cell strains
50Value is respectively 7.6 and 9.1ugml
-1(Cisplatin, DDP) IC50 to HL-60 is 8.7ml to the positive control cisplatin
-1The result shows that the anti-tumor activity of F-A is higher.
Table 1 F-A is to HL-60 and K
562Inhibitory action and IC
50Value
Drug level (ugml
-1) and suppression ratio % cell
IC
50(us·ml
-1)
50 25 12.5 6.25 3.125HL-60 93.0 71.2 57.3 46.7 44.4 7.6K
562 94.0 77.5 55.4 36.7 26.7 9.1
F-A influences the HL-60 cell under the F-A effect to the HL-60 cell growth curve, and growth is suppressed, and is concentration and time dependence (Fig. 1), and concentration is at 4ugml
-1The time can obviously suppress the growth of HL-60 cell.
F-A is right
3H-TdR mix test to influence F-A right after to HL-60 cytosis 3hr and 24hr
3The situation of mixing of H-TdR sees Table 2, and during F-A effect 3hr, the HL-60 cell is right
3It is little that H-TdR mixes influence, and behind the effect 24hr, right
3H-TdR is mixed with certain inhibitory action, is concentration dependent.Illustrate that F-A has certain synthetic effect of inhibition DNA.
Table 2 F-A mixes synthetic drug level CPM/5 * 10 that influence of HL-60 cell DNA to 3H-TdR
5Cell (suppression ratio (the %) (ugml of X ± SD)
-1) 3hr 24hr 3hr 24hr contrasts 4686 ± 1,091 12853.9 ± 2820.7--3.0 4824 ± 1,245 11832.5 ± 1002.3 0 7.910.0 3683.7 ± 242.9 7908 ± 1,201 21.4 38.550.0 3116 ± 726.7 7488 ± 231.5 33.5 41.7
F-A lacks the NBT reducing power to the human promyelocytic leukemia HL-60 cell that influences of HL-60 cell NBT reducing power, if cell is induced to the mature cell differentiation, then obtains the NBT reducing power.F-A is 4ugml respectively
-1And 8ugml
-1After acting on HL-60 cell 4d, the particulate positive cell rate of first occurs, compare P＞0.05, illustrate that F-A does not have differentiation-inducing action to the HL-60 cell with not administration group less than 5%.
[F-A is to the influence of HL-60 cell cycle distribution] F-A to HL-60 cytosis 8hr after the influence of cell cycle dna content see Table 3.
The cell cycle of table 3 F-A after to HL-60 cytosis 48hr changes
%G
0+ G
1%S %G
2+ M HL-60 cell contrasts 56.2 63.8 0 normal person's lymphocytes and contrasts 89.2 9.8 1.0 anticarcinogen 5-Fu and contrast 73.0 27.0 0 F-A (5ugml
-1) 47.3 36.2 16.4 F-A (10ugml
-1) 75.3 14.8 9.9
As table 3 result, not dosing group (matched group) HL-60 cell major part is in the S phase, and this is the characteristic (comparing with lymph cell) of tumor cell, does the anticarcinogen contrast with 5-Fu, and visible S phase cell obviously suppresses, and majority is G
1The phase cell.Illustrate that 5-Fu obviously suppresses the synthetic of DNA.The situation of F-A is different with 5-Fu, and it mainly shows G
2+ M phase cytosis, promptly this moment G
2+ M phase cell is obstructed, and makes cell stay in the M phase, illustrates that the effect of F-A may be relevant with the mitosis that suppresses the tumor cell.
As seen F-A influences transmissioning electric mirror checking to HL-60 cytohistology structure, and matched group oncocyte nuclear is big, irregular, nuclear/slurry ratio is big, and kernel is big, mostly is euchromatin in the nuclear, in the cell space high-visible (seeing Fig. 4 A, D, G) such as ribosome, endoplasmic reticulum and mitochondrions, treatment group (10ugml
-1) the oncocyte cell space is little, the oncocyte of visible a plurality of necrosis is examined for a short time, and nuclear/slurry ratio reduces, and nuclear concentrates, and heterochromosome is more, and is group's fast limit collection, and kernel diminishes, and is most destroyed.Organelle such as mitochondrion fog unclear.
3, conclusion
Studies show that the active anticancer of compound F 17-hydroxy-corticosterone-A is higher, to the IC of HL-60 cell 24hr
50At 5-7 μ gml
-1Between, the IC of 72hr
50Can reach 1 μ gml
-1, research finds that also its lethal effect mechanism to oncocyte may be relevant with the mitosis of cell, illustrates that F-A has certain novelty on chemical constitution and action principle, as a kind of new anti-cancer drug, have clinical value.
Embodiment two:
With preceding method F-B is studied, this chemical compound of also preliminary proof also has similar anti-tumor activity.
Embodiment three:
This research group has also adopted KB cell (CNE-2z) to carry out deep research, has confirmed that these two chemical compounds also have notable antitumor activity.
Embodiment four:
This research group entrusts department of pharmacy of Osaka, Japan university to adopt oral squamous carcinoma cell to study, and has proved that also these several chemical compounds have notable antitumor activity.
Embodiment five:
This research group studies show that further these two chemical compounds are to mice transplanting cancer EAC, S
180, tumor strain such as hepatocarcinoma has certain inhibitory action.
Embodiment six:
The preliminary proof of research, these two chemical compounds with influence that the synthetic chemotherapeutics fluorouracil of nucleic acid share or to form new preparation curative effect better.
Embodiment seven:
Above-mentioned two chemical compounds have been carried out the research of other pharmacology aspects, recognize that tentatively these two chemical compounds have certain antiinflammatory, anti rheumatism action and immunosuppressive action, are used for the treatment of various inflammation, rheumatism and various allergic disease clinically and have certain curative effect.
The pharmacodynamic study results suggest: this compounds can be formed compound preparation separately or with other relative medicines, comprises capsule, tablet, injection, electuary etc., provides clinical and is used for antitumor, antiinflammatory, rheumatism and treats various allergic diseases.
Claims (3)
1, the antineoplastic component that extracts from Pteridaceae plant Herba Pteridis Semipinnatae belongs to diterpene-kind compound, it is characterized in that containing α, β-methylene Ketocyclopentane structure, as following compound F 17-hydroxy-corticosterone-A or F-B:
Compound F 17-hydroxy-corticosterone-A:mp243-246 ℃, [α]
D-74 ° (c=0.2, MeOH)
7 α, 11 alpha-dihydroxy-s-15 oxygen-16 alkene-copy compound F 17-hydroxy-corticosterone-B:mp235-239 ℃ of sharp acid lactone (19,6), [α]
D-145 ° (c=0.2, MeOH)
11 α-15 oxygen-6 alkene-19 is copied sharp acid
2, medical composition as claimed in claim 1 is characterized in that these chemical compounds extract by following technology: clean air-dry with the Herba Pteridis Semipinnatae herb, ethanol extraction concentrates the back and goes up activated-charcoal column, with MeOH: CHCl
3(2: 1) eluting concentrates the back and goes up the silicagel column separation, with CHCl
3: MeOH (90: 2) eluting, collect respective components respectively and obtain.
3, medical composition as claimed in claim 1 is characterized in that these chemical compounds also can extract and separate from other Pteridaceae plants.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN95119380A CN1081920C (en) | 1995-12-12 | 1995-12-12 | Herba pteridis semipinnatae extract and its medicinal composite |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN95119380A CN1081920C (en) | 1995-12-12 | 1995-12-12 | Herba pteridis semipinnatae extract and its medicinal composite |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1138998A CN1138998A (en) | 1997-01-01 |
| CN1081920C true CN1081920C (en) | 2002-04-03 |
Family
ID=5081984
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN95119380A Expired - Fee Related CN1081920C (en) | 1995-12-12 | 1995-12-12 | Herba pteridis semipinnatae extract and its medicinal composite |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1081920C (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63146813A (en) * | 1986-12-11 | 1988-06-18 | Toa Nenryo Kogyo Kk | Carcinostatic agent containing diterpene based compound |
| JPS63146839A (en) * | 1986-12-11 | 1988-06-18 | Toa Nenryo Kogyo Kk | Carcinostatic agent containing pterosin |
-
1995
- 1995-12-12 CN CN95119380A patent/CN1081920C/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63146813A (en) * | 1986-12-11 | 1988-06-18 | Toa Nenryo Kogyo Kk | Carcinostatic agent containing diterpene based compound |
| JPS63146839A (en) * | 1986-12-11 | 1988-06-18 | Toa Nenryo Kogyo Kk | Carcinostatic agent containing pterosin |
Non-Patent Citations (1)
| Title |
|---|
| 《中药大辞典》，第1版 1986.1.1 江苏新医学辽，上海科学技术出版社 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1138998A (en) | 1997-01-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102018759B (en) | Rosmarinic acid, rosmarinic acid-containing common selfheal fruit-spike active ingredient and preparation methods and application thereof to prevention and treatment of cancer postoperative metastasis | |
| Gupta et al. | Use of flow cytometry to measure the immunostimulatory activity of aqueous extract of Jasminum auriculatum | |
| CN1081920C (en) | Herba pteridis semipinnatae extract and its medicinal composite | |
| CN102078317A (en) | Dihydroartemisinine-phospholipid complex and preparation and application thereof | |
| CN1775267A (en) | Kaikoujian extract, Its preparing method and use | |
| CN100336552C (en) | Medicine for treating lung cancer | |
| CN1126539C (en) | Application of tanshinone in preparing medicine for treating tumor | |
| CN109908188A (en) | A kind of Phellinus anti-cancer effective component extract and its preparation method and purposes | |
| CN112237588B (en) | Application of oyster mushroom polysaccharide selenoside-III anticancer active ingredient in preparation of medicine for resisting prostate cancer | |
| CN102614238A (en) | Application of lucide ganoderma and ginseng medicinal mycoplasm in preparing drugs for treating lung cancer | |
| CN112402413B (en) | Application of lindley eupatorium herb sesquiterpene lactone B in preparation of anti-liver cancer medicine and anti-liver cancer medicine | |
| CN1266149C (en) | Use of aspergillus fumigatus in antiflammation immunosuppressant | |
| CN100515461C (en) | Gamboge soft paste preparation | |
| CN1284535C (en) | Application of liganans of biphenyl cyelo-octadiene in preparing antineoplastic medicine | |
| CN1130213C (en) | Chinese herbal plaster for anti-cancer and analgesic | |
| CN111150752A (en) | Application of abrus herb extract in preparing anticancer medicine | |
| CN1883499A (en) | Drip pills of mangiferin and preparation method thereof | |
| CN109432084A (en) | A kind of anti-cancer composition and its application in medicine preparation | |
| CN113133994B (en) | Application of tricyclic diterpene natural product Vinigrol or pharmaceutical composition thereof in preparation of anti-cancer drugs | |
| CN115120585B (en) | Artemisinin B for resisting DDP drug-resistant lung adenocarcinoma and pharmaceutical composition thereof | |
| CN1194705C (en) | Natural toad skin for curing AIDS | |
| CN101099769B (en) | Application of Fructus Forsythiae in preparing anti-tumor chemotherapeutic sensitivity intensifying attenuating medicine | |
| CN110604758B (en) | Traditional Chinese medicine for relieving side effects of chemotherapy drugs | |
| Ristanti et al. | Immunological Responses in Female Rats Model with Anemia Treated with Snake Fruit Seed Flour Extract | |
| CN100410271C (en) | Compound with anti-tumor activity and its preparing method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |