CN101824014A - Compounds with anti-tumor activity in chloranthus japonicus as well as effective parts and purpose thereof - Google Patents
Compounds with anti-tumor activity in chloranthus japonicus as well as effective parts and purpose thereof Download PDFInfo
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Abstract
The invention discloses compounds with anti-tumor activity in chloranthus japonicus as well as effective parts and the purpose thereof. In the invention, three novel compounds with stronger anti-tumor activity are separated from the chloranthus japonicus which is a traditional Chinese medicine, i.e. a chloranthus japonicus element A, a chloranthus japonicus element B and a chloranthus japonicus element C. The method for separating the compounds comprises the following steps of: sequentially extracting a chloranthus japonicus extractive obtained by refluxing with 95% ethanol by using petroleum ether, ethyl acetate and n-butyl alcohol, and collecting the part obtained by ethyl acetate extraction; or degreasing the chloranthus japonicus extractive obtained by refluxing with 95% ethanol by using petroleum ether, carrying out macroporous resin column chromatograph, and eluting by using H2O, 50% alcohol and 95% alcohol sequentially; and carrying out silicagel column chromatography and ODS column chromatography on the ethyl acetate extracting part and the 50% alcohol component, and separating a prepared liquid phase to obtain the three compounds. Proved by in vitro anti-tumor activity research, the three compounds can remarkably restrain common tumor cells, have remarkable anti-tumor activity and can be used for developing novel anti-tumor harmfulless medicaments prepared from natural traditional Chinese medicines.
Description
Technical field
The present invention relates to three kinds of new compounds of a type sesquiterpene class, relate in particular to chloranthalactone type sesquiterpenoids of three kinds of tool anti-tumor activities that separation obtains in a kind of Chinese medicine Herba chloranthi japonici and preparation method thereof, the invention further relates to Herba chloranthi japonici anti-tumor effective component extract and preparation method thereof, the invention still further relates to this sesquiterpenoids and the Herba chloranthi japonici anti-tumor effective component extract medicinal use in the preparation antitumor drug, belong to Chinese medicine Herba chloranthi japonici extract field.
Background technology
After the eighties in 20th century, malignant tumour morbidity number is with average annual 3% to 5% speed increase, now occupied the primary dead cause of disease, endangered people's physical and mental health greatly, national economic development and social development have also been restricted, all caused white elephant for individual, family and society, so the research of anti-tumor medicine is very urgent.
At present, anti-tumor drug is mainly chemical synthetic drug both at home and abroad, though have certain curative effect, relatively poor to the long-term effect of tumour, and toxic side effect is bigger, and clinical application often is subjected to its strong toxic restriction.By comparison, the Chinese herbal treatment tumour embodies many-sided advantage, and with its special effect, little, the huge people's extensive concern that causes of exploitation potential of toxic side effect, the research of herbal medicine will be the effective way of antitumor drug exploitation.
Herba chloranthi japonici Chloranthus japonicus Sieb is the Chloranthaceae plant, have another name called yellow moccasin flower, beginning is stated from Shennong's Herbal, another name have solely shake grass, ghost solely shake grass, Four Heavenly Kings, four leaf gold, four leaves to, Herba Galii Bungei, Herba Chloranthi Henryi (Herba Choranthi seu Lysimachiae), Chinese enkianthus, leaflet celery, Su Ye wormwood artemisia, mountain rape, red bayberry grass, recklessly Macrophylla eye, Herba Chloranthi holostegii(Herba chloranthi japonici), Herba chloranthi japonici, four watts, four generation grass etc.Its herb hyoscine.The Herba chloranthi japonici medicinal history is long, and biological activity is stronger, and application among the people is very extensive.Flavor is hot, bitter, warm in nature, poisonous, has the clearing damp effects such as cold, promoting blood circulation and stopping pain, the stasis of blood detoxifcation of loosing of loosing, and cures mainly cough due to wind and cold, rheumatalgia, amenorrhoea, wound, swelling and pain due to blood stasis, hemoptysis etc.The wound that is used among the people, rheumatic arthralgia, common cold caused by wind-cold, pyogenic infections sore, the treatment of diseases such as venomous snake bite.Be distributed in provinces such as China Heilungkiang, Jilin, Hebei, Shanxi, Shandong, Shaanxi, Gansu.Its natural plant resource is very abundant.But so far, this plant is not almost carried out effective development and utilization.
Summary of the invention
One of the object of the invention provides a class and separate the chloranthalactone type sesquiterpenoids with anti-tumor activity that obtains from Herba chloranthi japonici;
Two of the object of the invention provides a kind of method of sesquiterpenoids of the chloranthalactone for preparing above-mentioned tool anti-tumor activity;
Three of the object of the invention is that the sesquiterpenoids with above-mentioned chloranthalactone is applied to prepare antitumor drug;
Four of the object of the invention provides Herba chloranthi japonici anti-tumor effective component and extracting method thereof;
Five of the object of the invention is that last Herba chloranthi japonici anti-tumor effective component is applied to prepare antitumor drug.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
The chloranthalactone type sesquiterpenoids of one class tool anti-tumor activity, it is the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici and the plain C of Herba chloranthi japonici, their structural formula is respectively with shown in following formula I, formula II and the formula III:
Formula II
The present invention is by the antitumor activity screening to Herba chloranthi japonici, find that the Herba chloranthi japonici general extractive has very strong restraining effect to tumour, find by active follow-up study, ethyl acetate extraction position in the general extractive or AB-8 macroporous adsorbent resin 50% ethanol elution position are anti-tumor effective component, to mouse S
180Sarcoma, mouse EAC ascites tumour and rat liver cancer H
22Noumenal tumour has stronger restraining effect, by chemical ingredients and biological activity follow, further determined the chloranthalactone type sesquiterpenoids chemical ingredients that has than powerful antitumor activity to the system of going deep into of the antineoplastic efficient part of Herba chloranthi japonici---the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain C of Herba chloranthi japonici.Above-mentioned three kinds of compounds have the common structure parent nucleus, be chloranthalactone type sesquiterpenoids chemical ingredients, difference called after: the plain A (formula I) (yinxiancaonin A) of Herba chloranthi japonici, chemical name: [(1 α, 3 α, 8 β)-and 6 β-hydroxy-1H-lindan-4,7 (11)-dien-4-formy-8,12-olide]; The plain B (formula II) (yinxiancaonin B) of Herba chloranthi japonici, chemical name: [(1 α, 3 α, 6 β, 8 β)-15-hydroxy-6,15-epoxy-1H-lindan-4,7 (11)-dien-olide]; The plain C (formula III) (yinxiancaonin C) of Herba chloranthi japonici, chemical name: [(1 α, 3 α, 8 β)-15-hydroxy-1H-lindan-4,7 (11)-dien-8,12-olide].
Above-mentioned three kinds of compounds can derive from the herb that former plant is Chloranthaceae plant Herba chloranthi japonici Chloranthus japonicus Sieb, also can derive from other plant of nearly edge of equal genus, also comprise any compound that adopts in three kinds of compounds of chemical complete synthesis approach acquisition.Also can directly obtain three kinds of new compounds by the extraction and separation method of chemistry, and the nearly edge kind of plant often having the common chemical ingredients, also is the effective way of the new biogenetic derivation of three kinds of new compounds of searching.In addition, development along with chemical synthesising technology, can directly obtain by complete synthesis approach for newfound active compound, more than be purpose to produce three kinds of new compounds, from former plant, directly extract or the application of the anti-tumor medical application by synthetic, semi-synthetic approach all falls into the scope of the invention.
As a reference, a kind of method of extracting from the herb of Chloranthaceae plant Herba chloranthi japonici, separating above-mentioned 3 kinds of compounds comprises the steps: the dry herb 95%EtOH refluxing extraction of Herba chloranthi japonici is reclaimed EtOH, obtains the EtOH extract; Again with behind the EtOH extract water suspendible, use sherwood oil, chloroform, ethyl acetate, n-butanol extraction successively respectively, obtain five components, each component is carried out the anti-tumor activity chase experiment, the result shows that the ethyl acetate extraction position has stronger anti-tumor activity, is antineoplastic efficient part; Continue to adopt separation methods such as silica gel column chromatography, ODS reversed-phase column chromatography and preparation HPLC this position to be separated to the ethyl acetate extraction position with technology, and differentiate and the parsing of various Wave Spectrum data through physics and chemistry, obtain the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain C of Herba chloranthi japonici respectively.
Of the present invention another prepare the method for above-mentioned three kinds of compounds, comprising: the Herba chloranthi japonici herb with after the 95%EtOH refluxing extraction, is obtained the EtOH extract; Again this extract is carried out AB-8 type macroporous resin column chromatography after with petroleum ether degreasing, use H successively respectively
2O, 50%EtOH, 95%EtOH wash-out obtain sherwood oil, H
2O, 50%EtOH, four parts of 95%EtOH prove by pharmacodynamic experiment, and 50%EtOH wash-out part also is the anti-tumor activity efficient part; Continue to adopt separation technology such as silica gel column chromatography, ODS reversed-phase column chromatography and HPLC to separate, also can obtain the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain C of Herba chloranthi japonici.
Continuing active the tracking shows, from anti-tumor effective component ethyl acetate extraction position or macroporous adsorbent resin column chromatography 50% ethanol elution position separate the plain A of the Herba chloranthi japonici obtain, the plain B of Herba chloranthi japonici, three kinds of new compounds of the plain C of Herba chloranthi japonici, has remarkable antitumor action, action intensity other chemical ingredients in the component can be used as antineoplaston medicine or prodrug.
Prove through external MTT anti-tumor experiment, the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain C of Herba chloranthi japonici all have the obvious suppression effect to various tumor cell strains such as human colon carcinoma LOVO, liver cancer HepG-2, prostate cancer PC-3, kidney OS-RC-2, ovarian cancer SKOV3, lung cancer A549, esophageal carcinoma Eca-109, cervical cancer He la, mammary cancer MCF-7, cancer of the stomach SGC-7901.Three kinds of compounds are 13.8～83.0 μ g/ml to the IC50 of kinds of tumor cell strain, illustrate that three kinds of compounds have stronger antitumor action; Though action effect is weaker than the 5 FU 5 fluorouracil group, side effect is little, and medication is safer; Each administration group suppresses effect and concentration and is dose-effect relationship in the finite concentration scope; Each administration group is being cultivated 72 hours tumor killing effect all significantly better than 48 hours inhibition effect; Each compound is better than action effect to other tumor cell lines to the inhibition effect of human colon carcinoma LOVO cell strain, liver cancer HepG-2.
The present invention also provides the medicinal use of the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain C of Herba chloranthi japonici, and its medicine is formed and can be made of following any composition: can be any of Herba chloranthi japonici plain A, B, C; Can be Herba chloranthi japonici plain A, B, C any two kinds; Can be Herba chloranthi japonici plain A, B, three kinds of common compositions of C; Also can directly directly make by the Herba chloranthi japonici anti-tumor effective component that comprises these three kinds of compositions; Perhaps by above various compositions as raw material, with the pharmaceutical composition of other kind compatibility of drugs, all can add pharmaceutically various pharmaceutical carriers of acceptable and auxiliary material again, be prepared from according to suitable technology and formulation.The pharmaceutical preparation, the Chinese medicine preparation that form with above-mentioned combination and compatibility all will fall into the present invention.
Any antineoplastic pharmaceutical compositions of the present invention can be prepared into it acceptable clinically injection formulations or oral preparations; Wherein, described injection formulations comprises injection, lyophilized injectable powder or infusion solution; Described oral preparations comprises tablet, granule, soft capsule, hard capsule or oral liquid and various slow control release type preparation.
Usually a kind of new compound can be used as prodrug or lead drug, finds new compound by structural modification and structure activity study; The new compound Herba chloranthi japonici of three kinds of anti-tumor activities of the present invention plain A, B, C can be used as prodrug; those skilled in the art pass through structural modification; on the basis of the common mother nucleus structure of the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici and the plain C of Herba chloranthi japonici; it is modified or transforms; the resultant on this basis a series of new compound that has identical parent nucleus and have anti-tumor activity, these new compounds and antineoplastic medicinal use thereof all should fall within protection scope of the present invention.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage of the present invention and characteristics will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall within the scope of protection of the present invention the details of technical solution of the present invention and form.
Embodiment 1
Take by weighing the dry herb 15kg of Herba chloranthi japonici, add 95% alcohol reflux of 10 times of amount volume ratios, extract altogether 2 times, each 2 hours, united extraction liquid reclaimed EtOH, obtains EtOH extract thick paste 1208g; With the water suspendible of this extract thick paste, obtain the water suspended matter again with 18 times of amount volume ratios; Use sherwood oil, chloroform, ethyl acetate, n-butanol extraction with water suspended matter equal volume amounts successively respectively, every kind of solvent extracts respectively 3 times, reclaims solvent, obtains sherwood oil, chloroform, ethyl acetate, propyl carbinol, five components of water; The active The selection result of following the tracks of proves that the ethyl acetate component is an anti-cancer effective component.Again the ethyl acetate component is carried out silica gel column chromatography, ODS reversed-phase column chromatography and preparation HPLC, obtain totally 12 of monomeric compounds, comprising three kinds of chloranthalactone type sesquiterpenoids new compounds: the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain C of Herba chloranthi japonici; To three kinds of compounds adopt UV spectrum (MeOH),
1H-NMR (CD
3OD) compose,
13The C-NMR spectrum,
1H-
1The integration analysis of H COSY spectrum, DEPT, HSQC and HMBC spectrum in conjunction with determination of physical constant etc., has been determined the structure of three kinds of compounds, and is new through looking into, and proves three kinds of new compounds, belongs to chloranthalactone type sesquiterpenoids chemical ingredients on its type of compounds.
Wherein, the plain A of Herba chloranthi japonici is white amorphous powder, is soluble in methyl alcohol, and UV spectrum (MeOH) maximum absorption occurs at the 222nm place, and molecular weight is 260, and molecular formula is C
15H
16O
4Process
1H-NMR (CD
3OD) compose,
13The C-NMR spectrum,
1H-
1The integration analysis of H COSY spectrum, DEPT, HSQC and HMBC spectrum is defined as (1 α, 3 α, 8 β)-and 6 β-hydroxy-1H-lindan-4,7 (11)-dien-4-formy-8,12-olide, be a kind of new compound, with the plain A (yinxiancaonin A) of its called after Herba chloranthi japonici.
The plain B of Herba chloranthi japonici is white amorphous powder, is soluble in methyl alcohol; UV spectrum (MeOH) maximum absorption occurs at the 221nm place, and molecular weight is 260, and molecular formula is C
15H
16O
4Process
1H-NMR (CD
3OD) compose,
13The C-NMR spectrum,
1H-
1The integration analysis of H COSY spectrum, DEPT, HSQC and HMBC spectrum is defined as (1 α, 3 α, 6 β, 8 β)-15-hydroxy-6,15-epoxy-1H-lindan-4,7 (11)-dien-olide are a kind of new compound, so with the plain B (yinxiancaoninB) of its called after Herba chloranthi japonici.
The plain C of Herba chloranthi japonici is white amorphous powder, is soluble in methyl alcohol, and UV spectrum (MeOH) maximum absorption occurs at the 224nm place, and molecular weight is 246, and molecular formula is C
15H
18O
3Process
1H-NMR (CD
3OD) compose,
13The C-NMR spectrum,
1H-
1The integration analysis of HCOSY spectrum, DEPT, HSQC and HMBC spectrum is defined as (1 α, 3 α, 8 β)-15-hydroxy-1H-lindan-4,7 (11)-dien-8, and 12-olide is a kind of new compound, the plain C (yinxiancaonin C) of called after Herba chloranthi japonici.
Embodiment 2
Take by weighing the dry herb 25kg of Herba chloranthi japonici, 95% alcohol reflux that adds 10 times of amount volume ratios, extract altogether 2 times, each 2 hours, united extraction liquid reclaims EtOH, obtains EtOH extract thick paste 2150g, again this extract thick paste is carried out AB-8 type macroporous resin column chromatography after with petroleum ether degreasing, use H successively respectively
2O, 50%EtOH, 95%EtOH wash-out obtain sherwood oil, H
2O, 50%EtOH, four parts of 95%EtOH.Prove that by the active tracking test of pharmacodynamics 50%EtOH partly is an efficient part.Continue to adopt separation technology such as silica gel column chromatography, ODS reversed-phase column chromatography and HPLC that this position is separated, and through the parsing of physico-chemical analysis and various Wave Spectrum data, be divided into from, identified the chemical structure of 5 chloranthalactone type sesquiterpenoids monomeric compounds, comprising new compound: the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain C of Herba chloranthi japonici.
The preparation of embodiment 3 slow releasing tablets, pill
Get the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain C monomeric compound of Herba chloranthi japonici, any array mode according to the three is that main ingredient is formed, also can directly form as main ingredient with the Herba chloranthi japonici efficient part, add right amount of auxiliary materials such as sustained release dosage, thinner, disintegrating agent, mixing is made particle, drying, compacting is in blocks, and dressing or spray film-coat are promptly; Perhaps add suitable matrix, drip and make the sustained-release dropping pill agent, be used for the antineoplaston purposes.
Embodiment 4 makes injection, infusion solution
Get the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain C monomeric compound of Herba chloranthi japonici, any array mode according to the three is that main ingredient is formed, and adds an amount of solubilizing agent, grinds, adding a small amount of water for injection again dilutes, mixing, it is an amount of to add sodium-chlor then, adds the injection water after the dissolving again to specified amount, filter, embedding, sterilization, promptly.
The plain A of test example 1 Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain three kinds of monomeric compound anticancer experiment in vitro of C (mtt assay) of Herba chloranthi japonici
One, specimen preparation
The plain A of three kinds of Herba chloranthi japonici, the plain B of Herba chloranthi japonici, the plain C compound monomer of Herba chloranthi japonici for this preparation of sample: get each monomeric compound to be measured respectively and dissolve with DMSO, the storing solution that is mixed with 2mg/ml with nutrient solution places-20 ℃ of preservations.Before facing usefulness, be diluted to final concentration with nutrient solution and be respectively: 2.5 μ g/ml, 5.0 μ g/ml, 10.0 μ g/ml, 25.0 μ g/ml, 50.0 μ g/ml and 100.0 μ g/ml.The DMSO final concentration is controlled at below 0.01% in wherein arbitrary dilution test liquid.
2. tumor cell line: with human colon carcinoma LOVO, liver cancer HepG-2, prostate cancer PC-3, kidney OS-RC-2, ovarian cancer SKOV3, lung cancer A549, esophageal carcinoma Eca-109, cervical cancer Hela, mammary cancer MCF-7, the strain of cancer of the stomach SGC-7901 attached cell with 0.25% trysinization after, be suspended in the RPMI1640 nutrient solution that contains 10% foetal calf serum, blow and beat into single cell suspension gently with the glass dropper, carry out cell counting, being diluted to concentration with nutrient solution is 20000 cell/ml.
3. positive control drug: get 5 FU 5 fluorouracil and dissolve with DMSO, the storing solution that is mixed with 2mg/ml with nutrient solution places-20 ℃ of preservations.Before facing usefulness, be diluted to final concentration with nutrient solution and be respectively: 0.1 μ g/ml, 1.0 μ g/ml, 10.0 μ g/ml, 100.0 μ g/ml.The DMSO final concentration is controlled at below 0.01% in wherein arbitrary dilution test liquid.
Two, test method
In each hole of 96 well culture plates, add the well-grown tumour cells of 100 μ l (about 2000 cells/well), after cultivation in 24 hours treats that cell is adherent fully, every hole adds 100 μ l need testing solutions respectively, every group parallel establishes six multiple holes, establish positive control simultaneously, negative control (adds substratum, cell, not dosing) and blank (only add substratum, do not add cell).In 37 ℃, 5%C0
2Cultivate 48h, 72 hours under the condition respectively.Then, every hole adds MTT liquid 20 μ l (5mg/L), continues to cultivate 4h, discards nutrient solution, and every hole adds 100 μ l DMSO, after the dissolving fully to be crystallized, surveys absorbance, i.e. OD value in the 490nm place with microplate reader.Calculate cell proliferation inhibition rate.Every group of trial-product repeated experiments three times averaged.
Cell proliferation inhibition rate=100%-cell survival rate;
IC
50: inhibition rate of tumor cell is the mass concentration of 50% o'clock medicine.
Three, test-results
Negative control, the cell well-grown, no growth-inhibiting phenomenon illustrates that test method is feasible, the IC that the dosing group is measured
50Value sees Table 1:
Table 1 Herba chloranthi japonici plain A, B, C extracorporeal suppression tumor cell strain effect
Four, conclusion (of pressure testing)
Three kinds of compounds are 13.8～83.0 μ g/ml to the IC50 of kinds of tumor cell strain, illustrate that three kinds of compounds have stronger antitumor action; Though action effect is weaker than the 5 FU 5 fluorouracil group, side effect is little, and medication is safer; Each administration group suppresses effect and concentration and is dose-effect relationship in the finite concentration scope; Each administration group is being cultivated 72 hours tumor killing effect all significantly better than 48 hours inhibition effect; Each compound is better than action effect to other tumor cell lines to the inhibition effect of human colon carcinoma LOVO cell strain, liver cancer HepG-2.
The different extract part anti-tumor in vivo experiments of test example 2 Herba chloranthi japonici
One, test materials
1. test sample preparation: get Herba chloranthi japonici and reflux 2 times with the 95%EtOH of 10 times of volumes, each 1 hour, filtration, merging filtrate, decompression and solvent recovery obtains general extractive; General extractive is suspended in water the back with petroleum ether extraction, obtain petroleum ether extract; Continue it, adopt macroporous adsorbent resin column chromatography, use H successively respectively
2O, 50%, 95%EtOH carry out wash-out, obtain corresponding H respectively
2O, 50%, 95%EtOH eluate.
Endoxan (powder injection), Hengrui Medicine Co., Ltd., Jiangsu Prov., specification 200mg/ bottle, lot number 060321.
2. laboratory animal Kunming kind small white mouse, male and female have both, and body weight 22 ± 2g, conformity certification number are: P00102006 number, provided by Heilongjiang University of Chinese Medicine's Experimental Animal Center
Murine sarcoma S
180Knurl strain, rat liver cancer H
22Knurl strain, the strain of mouse ascites EAC knurl: introduce by Harbin Medical University institute of oncology, go down to posterity with ascitic type.
Experimental animal model is set up: aseptic condition extracts inoculation 7 days, well-grown S down
180, H
22And EAC tumor-bearing mice ascites, with sterile saline (＜1: 3) dilute, press only subcutaneous or abdominal injection ascites of 0.1ml/, promptly about 5 * 10 through right armpit
6Individual oncocyte, living cell rate＞95% is made solid tumor type or ascitic type bearing mouse model.
Two, test method
Blank group, endoxan group, general extractive group, petroleum ether extract group and H are established in this test
2O, 50%, 95%EtOH eluate group, each experimental group is by amounting to the administration of identical crude drug amount (because of there is difference in the productive rate of each component, so the dosage of each component is amounted to into identical crude drug amount, to indicate under the identical crude drug amount situation, the power of each component antitumor action), irritate stomach, the blank group is irritated stomach (0.2mL/10g) with 0.9% physiological saline, positive controls endoxan abdominal injection (0.1mL/10g), solid tumor type mouse is random packet after inoculating 24 hours, and successive administration was weighed, put to death after 8 days, got knurl respectively, claim knurl heavy, calculate tumour inhibiting rate; Ascitic tumor type mouse is random packet after inoculating 24 hours, and successive administration 8 days is observed also record dead mouse situation every day.
Three, test-results
1. the different separated parts of Herba chloranthi japonici are to mouse S
180The influence of sarcoma
The results are shown in Table 2:
The different separated parts of table 2 Herba chloranthi japonici are to mouse S
180The influence of sarcoma (X ± S)
Annotate: compare * * P＜0.01 with the blank group
As seen from Table 2, ether extract, water elution thing are to mouse S
180The sarcoma restraining effect a little less than, its inhibiting rate is compared there was no significant difference with the blank group; 50% pure eluate is to mouse S
180The inhibiting rate of sarcoma is 55.5%, with the blank group utmost point significant difference (P＜0.01) is arranged, and antitumor activity and general extractive compare, a little less than the general extractive group, but there was no significant difference (P＞0.05), so 50% pure eluate is anti-mouse S
180The sarcoma reactive site.
2. the different separated parts of Herba chloranthi japonici are to rat liver cancer H
22The influence of solid tumor
Test-results sees the following form 3:
The different separated parts of table 3 are to rat liver cancer H
22The influence of solid tumor (X ± S)
Annotate: compare * * P＜0.01 with the blank group
As can be known from the above table, ether extract, water elution thing are to rat liver cancer H
22The solid tumor restraining effect a little less than, compare there was no significant difference with the blank group.50% pure eluate is to rat liver cancer H
22The inhibiting rate of solid tumor is 53.0%, has compared utmost point significant difference (P＜0.01) with the blank group, though antitumor activity a little less than the general extractive group, there was no significant difference (P＞0.05) is anti-rat liver cancer H
22The solid tumor reactive site.
3. the different separated parts of Herba chloranthi japonici are to the influence of EAC ascitic tumor mouse survival time
Test-results sees Table 4:
The different separated parts of table 4 are to the influence of EAC ascitic tumor mouse survival time (X ± S)
Annotate: compare * * P＜0.01 with the blank group
As known from Table 4, ether extract, water elution thing, 95% pure eluate influence less to EAC ascitic tumor mouse survival time, compare there was no significant difference with the blank group; 50% pure eluate oral administration is compared with the blank group, there is tangible prolongation effect the life survival time to EAC ascitic tumor mouse, the rate elongation rate is 37.3%, compare with blank that there were significant differences (P＜0.01), compare no significant difference (P＞0.05) with the endoxan group, can be considered prolongation EAC ascitic tumor mouse survival time reactive site.
Four, conclusion (of pressure testing)
50% elution fraction has stronger anti-tumor activity, and the tumour of S180 sarcoma mouse, liver cancer H22 solid tumor mouse is all had obvious restraining effect, can obviously prolong the survival time of EAC ascitic tumor mouse, is antineoplastic efficient part.Further under anti-tumor activity is followed the trail of, study the chemical ingredients of efficient part, seeking the strong monomeric compound of anti-tumor activity is highly significant.
Claims (10)
1. have three new compounds of anti-tumor activity, it is characterized in that: described compound is a chloranthalactone type sesquiterpenoids.
2. according to the described compound of claim 1, it is characterized in that: it is the plain A of Herba chloranthi japonici, the plain B of Herba chloranthi japonici and the plain C of Herba chloranthi japonici, and their structural formula is respectively with shown in following formula I, formula II and the formula III:
3. a method for preparing the described compound of claim 2 comprises: with the dry herb 95%EtOH refluxing extraction of Herba chloranthi japonici, reclaim EtOH, obtain the EtOH extract; With behind the EtOH extract water suspendible, use sherwood oil, chloroform, ethyl acetate, n-butanol extraction successively respectively again, obtain five components respectively; Continue to adopt silica gel column chromatography, ODS reversed-phase column chromatography or preparation HPLC method that the ethyl acetate component is separated to the ethyl acetate component, the parsing through physics and chemistry discriminating and various Wave Spectrum data obtains compound shown in formula I, formula II and the formula III respectively.
4. a method for preparing the described compound of claim 2 comprises: after the Herba chloranthi japonici herb is used the 95%EtOH refluxing extraction, obtain the EtOH extract; Again the EtOH extract is carried out AB-8 type macroporous resin column chromatography after with petroleum ether degreasing, use H successively respectively
2O, 50%EtOH, 95%EtOH wash-out are collected 50%EtOH wash-out part; Partly adopt silica gel column chromatography, ODS reversed-phase column chromatography or HPLC method to separate the 50%EtOH wash-out, obtain compound shown in formula I, formula II and the formula III respectively.
5. Herba chloranthi japonici anti-tumor effective component extract, its preparation method comprises: with the dry herb 95%EtOH refluxing extraction of Herba chloranthi japonici, recovery EtOH obtains the EtOH extract; With behind the EtOH extract water suspendible, use sherwood oil, chloroform, ethyl acetate, n-butanol extraction successively respectively again, collect the ethyl acetate extraction position, promptly.
6. Herba chloranthi japonici anti-tumor effective component extract, its preparation method comprises: with the Herba chloranthi japonici herb with the 95%EtOH refluxing extraction after, obtain the EtOH extract; Again the EtOH extract is carried out AB-8 type macroporous resin column chromatography after with petroleum ether degreasing, use H successively respectively
2O, 50%EtOH, 95%EtOH wash-out are collected 50%EtOH wash-out part, promptly.
7. claim 1 or 2 described compounds are in the purposes of preparation in the antitumor drug.
8. claim 5 or 6 described effective part extracts are in the purposes of preparation in the antitumor drug.
9. an antineoplastic pharmaceutical compositions is characterized in that: comprise claim 1 or 2 described 1-3 kind compounds and/or the claim 5 or the 6 described effective part extracts of significant quantity, add pharmaceutically acceptable carrier or auxiliary material and form.
10. according to the described antineoplastic pharmaceutical compositions of claim 9, it is characterized in that: it is prepared into acceptable clinically injection formulations or oral preparations; Wherein, described injection formulations comprises injection, lyophilized injectable powder or infusion solution; Described oral preparations comprises tablet, granule, soft capsule, hard capsule, oral liquid or slow control release type preparation.
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| CN102516213A (en) * | 2011-11-30 | 2012-06-27 | 浙江大学 | Sesquiterpene compound and preparation method and application thereof |
| CN103127063A (en) * | 2011-11-28 | 2013-06-05 | 复旦大学 | Application of chloranthus japonicus alcohol F in preparation of antitumor drugs |
| CN103127060A (en) * | 2011-11-28 | 2013-06-05 | 复旦大学 | Application of chloranthus japonicus alcohol D in preparation of antitumor drugs |
| CN105106276A (en) * | 2015-09-23 | 2015-12-02 | 刘高志 | Tradescantia extract as well as preparation method and medical application of tradescantia extract |
| CN105193887A (en) * | 2015-09-09 | 2015-12-30 | 刘高志 | Acanthopanax senticosus extract and antioxidation application thereof |
| CN114773304A (en) * | 2022-04-08 | 2022-07-22 | 南开大学 | Linderane type sesquiterpene compound separated from chloranthus japonicus extract and application thereof in preparing medicine for treating liver cancer |
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2010
- 2010-02-11 CN CN2010101086285A patent/CN101824014B/en active Active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103127063A (en) * | 2011-11-28 | 2013-06-05 | 复旦大学 | Application of chloranthus japonicus alcohol F in preparation of antitumor drugs |
| CN103127060A (en) * | 2011-11-28 | 2013-06-05 | 复旦大学 | Application of chloranthus japonicus alcohol D in preparation of antitumor drugs |
| CN102516213A (en) * | 2011-11-30 | 2012-06-27 | 浙江大学 | Sesquiterpene compound and preparation method and application thereof |
| CN105193887A (en) * | 2015-09-09 | 2015-12-30 | 刘高志 | Acanthopanax senticosus extract and antioxidation application thereof |
| CN105106276A (en) * | 2015-09-23 | 2015-12-02 | 刘高志 | Tradescantia extract as well as preparation method and medical application of tradescantia extract |
| CN114773304A (en) * | 2022-04-08 | 2022-07-22 | 南开大学 | Linderane type sesquiterpene compound separated from chloranthus japonicus extract and application thereof in preparing medicine for treating liver cancer |
| CN114773304B (en) * | 2022-04-08 | 2024-04-12 | 南开大学 | Linderane type sesquiterpene compound separated from herba Lespedezae Cuneatae extract and its application in preparing medicine for treating liver cancer |
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