CN102579425A - Caulis Spatholobi extract, application thereof and new application of isoliquiritigenin - Google Patents
Caulis Spatholobi extract, application thereof and new application of isoliquiritigenin Download PDFInfo
- Publication number
- CN102579425A CN102579425A CN2012100196959A CN201210019695A CN102579425A CN 102579425 A CN102579425 A CN 102579425A CN 2012100196959 A CN2012100196959 A CN 2012100196959A CN 201210019695 A CN201210019695 A CN 201210019695A CN 102579425 A CN102579425 A CN 102579425A
- Authority
- CN
- China
- Prior art keywords
- extract
- isoliquiritigenin
- ethanol
- caulis spatholobi
- extraction
- 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
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides Caulis Spatholobi extract, an application thereof and a new application of isoliquiritigenin, and particularly provides Caulis Spatholobi extract containing isoliquiritigenin, an active part and a monomer compound isoliquiritigenin as well as an application of a composition in preparing medicine for treating and preventing triple-negative breast cancer. The composition consists of isoliquiritigenin and one or more active ingredients in gallocatechin, epicatechin and catechin.
Description
Technical field
The present invention relates to the new purposes of Caulis Spatholobi extract and isoliquiritigenin, Caulis Spatholobi extract and the isoliquiritigenin that is specifically related to contain isoliquiritigenin is in the preparation prevention and treat the application in the three negative breast cancer medicines.
Background technology
Caulis Spatholobi is the dry rattan that pulse family (Leguminosae) spatholobus suberectus belongs to (Spatholobus) plant spatholobus suberectus (Spatholobus suberectus Dunn).Be the nourshing blood and promoting blood circulation effect that Song Yihou finds, a spot of bibliographical information is arranged in recent years, this medicine has antitumor action, and we carry out systematic research to this medicine.
In recent years discover that Caulis Spatholobi and extract thereof have the antineoplastic activity, but to breast carcinoma, particularly three negative breast carcinomas (the type breast carcinoma that this is special) are not appeared in the newspapers.
As everyone knows, breast carcinoma is the special relatively tumor of a quasi-biology characteristic, is one of modal malignant tumor of women, and its sickness rate is high, serious threat women's psychosomatic health.According to the dependence situation of tumor to hormone; Detect back estrogen receptor (estrogen receptor through SABC; ER), progesterone receptor (progesterone receptor, PR) and human epidermal growth factor receptor 2 (HER2) all be expressed as negative patient with breast cancer owing to lack special targeted therapy, prognosis is relatively poor; This be three negative breast cancer (triple negative breast cancer, TNBC).
Statistical datas such as Anders show that annual about 1,000,000 women in the whole world are diagnosed as breast carcinoma, and wherein about 17% is three negative breast cancer.Sickness rate that there are some researches show African American Women's three negative breast cancer is about 2 times of pink toes; Three negative breast cancer ratios of Korea S's report are 14.7%; Japan is 15%; China does not still have the correlational study data report of multicenter, large sample at present, but reports that its ratio should be lower than black race and be higher than white people.Interethnic morbidity difference is that gene and sudden change thereof are the basic reasons that causes these differences.
Three negative breast cancer are characteristic with invasive clinical manifestation, compare with the other types breast carcinoma, and relatively poor total survival rate and DFS phase are arranged; The axillary lymphatic metastasis rate is low, and the generation that internal organs (like lung) shift early; Local relapse is higher, the risk of relapse 1-3 after treatment that mostly occurs.Aspect clinical pathology, three negative breast cancer have that diameter of tumor is big, tumor cell exist more central authorities downright bad, higher proliferation activity arranged.
This medicine of discovering in early stage can suppress lactic acid dehydrogenase-A (LDH-A; It is the key enzyme of final step in the glycolytic pathway; It is not expressed in the competent normal cell of oxygen supply basically; Yet in various tumor cells, because the influence of its mitochondrial function defective and anoxia microenvironment, LDH-A expresses obviously to be increased).The LDH-A down-regulated expression, LDH-A is active to be reduced, and can effectively stop aerobic glycolysis, let tumor can't obtain energy, thereby dodar suppresses the effect of tumor.
In view of breast carcinoma to carbohydrate metabolism by way of special dependency, suppressing aerobic glycolysis has become one of important channel that suppresses breast cancer cell, and becomes one of new targeting important means of anti-breast carcinoma.Suppress the New Policy that glycolysis can be used as the breast cancer tumor treatment.
Summary of the invention
One object of the present invention is to provide a kind of Caulis Spatholobi extract and active site that contains isoliquiritigenin to prevent and treat the application in the three negative breast cancer medicines in preparation;
Another object of the present invention is to provide isoliquiritigenin to prevent and treat the application in the three negative breast cancer medicines in preparation.
Another object of the present invention is to provide a kind of pharmaceutical composition of treatment three negative breast cancer.
Another object of the present invention is to provide a kind of preparation of pharmaceutical compositions to treat the application in the three negative breast cancer medicines.
Caulis Spatholobi extract according to the invention is Caulis Spatholobi water extract or ethanol extraction;
The preferred 60-120 of said Caulis Spatholobi water extract ℃ water extraction 1-8 time; More preferably extract 2-5 time;
Said Caulis Spatholobi ethanol extraction, preferred 10-95% ethanol extraction, more preferably 20-80% ethanol extraction, more preferably 30-70% ethanol extraction, more preferably 40-70% ethanol extraction; Extracting mode can select to adopt percolation, backflow, dipping or supersound extraction; The preferred extraction 1-8 time; More preferably extract 2-5 time;
The preferred above-mentioned Caulis Spatholobi water extract of Caulis Spatholobi extract according to the invention, ethanol extraction successively extract or select the extract that an extraction obtains successively with petroleum ether, ethyl acetate, n-butyl alcohol;
The further preferred macroporous resin of going up of the extract that obtains of extraction use earlier water elution, after the ethanol liquid eluting that increases gradually with concentration successively, the eluate of collection high concentration ethanol is the Caulis Spatholobi active site;
The present invention provides a kind of new Caulis Spatholobi extract that contains isoliquiritigenin; This extract is prepared by following method: use n-butanol extraction behind Caulis Spatholobi water extraction or the ethanol extraction, extract is macroporous resin on conventional method, uses earlier water elution; The ethanol liquid eluting that the back increases with concentration in 10-95% concentration of alcohol scope successively gradually; Comprising using the 60-90% ethanol elution, collect 60-90% ethanol elution thing, promptly get;
Preferably, said macroporous resin is a low pole, further is preferably the D101 macroporous resin;
The preferred 60-120 of said Caulis Spatholobi water extraction ℃ water extraction 1-8 time; More preferably extract 2-5 time;
Said Caulis Spatholobi ethanol extraction, preferred 10-95% ethanol extraction, more preferably 20-80% ethanol extraction, more preferably 30-70% ethanol extraction, more preferably 40-70% ethanol extraction; Extracting mode can select to adopt percolation, backflow, dipping or supersound extraction; The preferred extraction 1-8 time; More preferably extract 2-5 time;
The method for preparing of Caulis Spatholobi active site according to the invention is preferably: Caulis Spatholobi water extract or ethanol extract dispersing and dissolving in water, are used n-butanol extraction; Butanol extraction liquid gets n-butyl alcohol extract after being evaporated to and doing, and n-butyl alcohol extract adsorbs through the D101 macroporous adsorptive resins; Wash to eluent colourless after, use 15-25%, 3545%, 60-90% alcoholic solution eluting successively, collect 80% ethanol elution thing; Be evaporated to driedly, promptly get.More preferably comprise 80% ethanol elution, collect 80% ethanol elution thing.
The present invention further discloses isoliquiritigenin and prevents and treat the application in the three negative breast cancer medicines in preparation;
Said isoliquiritigenin can be bought from market; Also can from Caulis Spatholobi, extract and obtain; Can also from other plant, separation and purification obtain; Also can obtain through chemosynthesis or biological synthesis method, its structural formula is following: the present invention provides a kind of pharmaceutical composition that prevents or treat three negative breast cancer, and the active component of said composition mainly is made up of one or more active component in isoliquiritigenin and optional nutgall catechin, epicatechin and the catechin.
Said pharmaceutical composition, one or more active component that preferably reached in optional nutgall catechin 5-15 weight portion, epicatechin 150-250 weight portion, the catechin 150-250 weight portion by isoliquiritigenin 1-9 weight portion are formed.
In the said compositions, preferably reach one or more active component of choosing nutgall catechin 8-12 weight portion, epicatechin 180-230 weight portion, catechin 180-230 weight portion wantonly and form by isoliquiritigenin 3-7 weight portion.
Pharmaceutical composition of the present invention, preferably form by following active component:
Isoliquiritigenin 1-9 weight portion nutgall catechin 5-15 weight portion
Epicatechin 150-250 weight portion.
Pharmaceutical composition of the present invention, preferably form by following active component:
Isoliquiritigenin 3-7 weight portion nutgall catechin 8-12 weight portion
Epicatechin 180-230 weight portion.
Pharmaceutical composition of the present invention, more preferably form by following active component:
Isoliquiritigenin 5 weight portion nutgall catechins 10 weight portions
Epicatechin 200 weight portions.
Pharmaceutical composition of the present invention, preferably form by following active component:
Isoliquiritigenin 1-9 weight portion nutgall catechin 5-15 weight portion
Epicatechin 150-250 weight portion catechin 150-250 weight portion;
Pharmaceutical composition of the present invention, preferably form by following active component:
Isoliquiritigenin 3-7 weight portion nutgall catechin 8-12 weight portion
Epicatechin 180-230 weight portion catechin 180-230 weight portion.
Pharmaceutical composition of the present invention, more preferably form by following active component:
Isoliquiritigenin 5 weight portion nutgall catechins 10 weight portions
Epicatechin 200 weight portion catechins 200 weight portions.
Isoliquiritigenin according to the invention, nutgall catechin, epicatechin and catechin can be extracted from Caulis Spatholobi and obtain, and can also from other plant, separation and purification obtain, and also can obtain through chemosynthesis or biological synthesis method.
Caulis Spatholobi extract of the present invention, active site, monomeric compound and compositions can add conventional adjuvant according to common process and process the acceptable any preparation of pharmaceutics, like medicinal tea, capsule, tablet, granule, gel, slow releasing agent, oral liquid, drop pill or nanometer formulation; Also can use, process corresponding preparation with other related drugs compatibilities.
Experimental example
Experimental example 1 Caulis Spatholobi active site of the present invention and the research of monomeric compound inside and outside antitumor
1, the experiment in vitro of Caulis Spatholobi active site and monomeric compound
1.1 experimental drug: 80% eluate (making) by embodiment 4, isoliquiritigenin (Isoliquiritigenin) (purchasing) in Nat'l Pharmaceutical & Biological Products Control Institute,
1.2 experimental technique: the cell MCF-7 of the trophophase of taking the logarithm respectively (cell strain that estrogen receptor relies on); MDA-MB-231 (three negative breast cancer cell strains; Be the cell strain that non-estrogen receptor relies on) and MCF-10 (matched group; The normal mammary glandular cell strain of people) every hole 100 μ L are inoculated in 96 orifice plates, and cell concentration is 5 * 10
4/ mL; After cultivation 24h treats cell attachment; Change serum-free medium; Absorb old culture medium behind the 24h, add the medicinal liquid (containing 2 ‰ DMSO) that 90 μ L serum-free mediums add Caulis Spatholobi 80% eluate (SS) 10-100 μ g/mL and isoliquiritigenin (ISO) 20-80uM more respectively, make the medicinal liquid final concentration be respectively 10-100 μ g/mL; 0-80uM; The blank group adds the phosphate buffer that 10 μ L contain 2 ‰ DMSO, establishes 6 parallel holes for every group, places 37 ℃, 5%CO
2Cultivate 48h in the cell culture incubator.Collecting cell is measured the inhibition of medicine cell growth, the influence of apoptosis rate.
1.3 experimental result: see table 1,2.
The 80% pure eluate of table 1. Caulis Spatholobi, isoliquiritigenin are to the influence of two kinds of growth of tumour cell
Experimental result: carry out cell culture with 80% eluate and isoliquiritigenin (ISO), observation of cell growth rate, result show that the two all has the effect that suppresses tumor growth, and have the relation (promptly with the increase of administration agent amount, curative effect increases) that dose-effect relies on.Isoliquiritigenin (ISO) compares the better effects if of ISO with 80% eluate.Inhibition effect to three negative cells strains is superior to the cell strain to the estrogen dependence.
The 80% pure eluate of table 2. Caulis Spatholobi, isoliquiritigenin are to the influence of two kinds of apoptosis of tumor cells rates
Experimental result shows that 80% eluate and isoliquiritigenin (ISO) all have the effect that promotes apoptosis of tumor cells, and has the relation (promptly with the increase of administration agent amount, curative effect increases) that dose-effect relies on.ISO and 80% eluate are compared, the better effects if of ISO.And the two inhibition effect to three negative cells strains is superior to the cell strain that estrogen relies on.
2, Caulis Spatholobi active site and isoliquiritigenin are to the influence of two types of different tumor cell line protein expressions
2.1 experimental drug: 80% eluate (making), isoliquiritigenin (purchasing) in Nat'l Pharmaceutical & Biological Products Control Institute by embodiment 4
2.2 experimental technique: each cell MCF-7 of the trophophase of taking the logarithm respectively, the every hole 100 μ L of MDA-MB-231 are inoculated in 96 orifice plates, and cell concentration is 5 * 10
4/ mL; After cultivation 24h treats cell attachment; Change serum-free medium; Absorb old culture medium behind the 24h, add the medicinal liquid (containing 2 ‰ DMSO) that 90 μ L serum-free mediums add Caulis Spatholobi 80% eluate (SS) 20-80 μ g/mL and isoliquiritigenin (ISO) 20-80uM more respectively, make the medicinal liquid final concentration be respectively 10-10 μ g/mL; 0-80uM; Place 37 ℃, 5%CO
2Cultivate 48h in the cell culture incubator.Collecting cell extracts albumen, measures the influence of medicine to the tumor cell protein expression with Western blot method.
2.3 experimental result: see Fig. 1,2.Experimental result shows that 80% eluate and isoliquiritigenin can pass through inducing cell mitochondrion approach apoptosis, has tangible dose-effect dependence, thus the effect of the anti-two types of breast tumor of performance.
3, suppress the effect of tumor growth in the body
3.1 experimental drug: 80% eluate (making), isoliquiritigenin (purchasing) in Nat'l Pharmaceutical & Biological Products Control Institute by embodiment 4
3.2 experimental technique: adopt the method for MDA-MB-231 and MCF-7 tumor cell transplantation to set up breast carcinoma nude mice animal model.At the position of mammary gland injection 3*10
6, dosage: 80% eluate 5mg/kg, isoliquiritigenin low dose group 20ug/kg, isoliquiritigenin high dose group 40ug/kg.Observe relatively each drug group and the tumor size of negative blank control group nude mice and the variation of weight.After experiment finishes, collect the tumor of each animal groups, observe the relatively interior influence that tumor growth is suppressed of Caulis Spatholobi extract body.
3.3 experimental result
The influence that in the 80% pure eluate (SS) of table 3 Caulis Spatholobi, the isoliquiritigenin body tumor growth is suppressed
The result shows; 80% eluate, isoliquiritigenin are grown in vivo to two types breast carcinoma and are all presented inhibitory action; And the inhibition effect of MDA-MS-231 slightly is superior to better effects if that MCF-7 explains that promptly the inhibitory action to three negative breast carcinomas relies on than estrogen receptor, and (rate of increase MDA-MS-231 of its tumor is lower relatively, tumor proliferation rate=RTVT/RTVC * 100%.RTVT=administration group relative tumour volume; RTVC=matched group relative tumour volume).Isoliquiritigenin is strong to the ability of neoplasm growth than 80% eluate, and apoptosis rate is also than the eluting object height.
Experimental example 2 Caulis Spatholobi extracts of the present invention, active site and monomeric compound body are to the influence of LDH
1, Caulis Spatholobi extract and monomeric compound thereof suppress the effect of the expression of breast tumor cell LDH-A
1.1 experimental drug: Caulis Spatholobi water extract (H
2O) (make) ethanol extract (60%EtOH) (making), isoliquiritigenin (purchasing) in Nat'l Pharmaceutical & Biological Products Control Institute by embodiment 2 by embodiment 1;
Different dosage form solvent extract: Caulis Spatholobi medicinal material coarse powder 1000g; With 60% ethanol ultrasonic extraction of 10 times of amounts (W/V) 2 times; Each 1 hour, merge extracted twice liquid, be evaporated to do not have the alcohol flavor after; Extract successively with petroleum ether, ethyl acetate, n-butyl alcohol respectively, obtain 4 extracts that polarity is different: petroleum ether extract (PE) 1.1g, acetic acid ethyl ester extract (EtOAC) 11.5g, n-butyl alcohol extract (BUOH) 24.4g and water layer stay excess (DW) 79.3g.
1.2 experimental technique: the MCF-7 of the trophophase of taking the logarithm respectively, the every hole 100 μ L of MDA-MB-231 and MCF-10 are inoculated in 96 orifice plates, and cell concentration is 5 * 10
4/ mL; After cultivation 24h treats cell attachment under normal pressure and the anoxia condition; Change serum-free medium, absorb old culture medium behind the 24h, add the medicinal liquid (containing 2 ‰ DMSO) that 90 μ L serum-free mediums add Caulis Spatholobi water extract, ethanol extract, different solvents extract 10-100 μ g/mL and isoliquiritigenin 10-50uM respectively more respectively; Make the medicinal liquid final concentration be respectively 10-100 μ g/mL, 10-50uM; The blank group adds the phosphate buffer that 10 μ L contain 2 ‰ DMSO, establishes 6 parallel holes for every group, places 37 ℃, 5%CO
2Cultivate 48h in the cell culture incubator.Behind the collecting cell, extract total protein, be used to measure the influence of medicine pair cell LDH-A activity, protein expression.Apoptosis rate with its cell of cells were tested by flow cytometry.
1.3 experimental result: see table 4.
Active (U/mg) (dosage is 50ug/ml) of the different extract pair cell of table 4 LDH and apoptotic influence
Experimental result shows n-butyl alcohol, and acetic acid ethyl ester extract and water layer stay the inhibitory action of LDH-A of excess better, but water extract, ethanol extract also have certain effect, but the having better effect of n-butanol extraction position.Also explanation simultaneously, the observation different pharmaceutical also is the best results at n-butanol extraction position to the apoptosis rate of inducing tumor cell.
2,80% eluate and isoliquiritigenin under normal oxygen and anoxia bar to the active influence of breast tumor cell LDH-A
1.1 experimental drug: 80% eluate (making), isoliquiritigenin (purchasing) in Nat'l Pharmaceutical & Biological Products Control Institute by embodiment 4.
1.2 experimental technique: because the generation of LDH-A produces under anaerobic environment usually, tumor tissues is exactly an anaerobic environment, so with the influence to the LDH-A expression under often oxygen and anaerobic environment of 80% eluate and isoliquiritigenin.Take the logarithm the respectively MCF-7 of trophophase, the every hole 100 μ L of MDA-MB-231 are inoculated in 96 orifice plates, and cell concentration is 5 * 10
4/ mL; After cultivation 24h treats cell attachment under normal pressure and the anoxia condition; Change serum-free medium, absorb old culture medium behind the 24h, add the medicinal liquid (containing 2 ‰ DMSO) that 90 μ L serum-free mediums add Caulis Spatholobi 80% eluate 10-10 μ g/mL and isoliquiritigenin 10-50uM more respectively; Make the medicinal liquid final concentration be respectively 10-100 μ g/mL, 10-50uM; The blank group adds the phosphate buffer that 10 μ L contain 2 ‰ DMSO, establishes 6 parallel holes for every group, places 37 ℃, 5%CO
2Cultivate 48h in the cell culture incubator.Behind the collecting cell, extract total protein, it is active to be used to measure medicine pair cell DH-A.
1.3 experimental result: see table 5,6.
Table 5 80% eluate is in the influence (uM concentration) to LDH-A of normal oxygen and anaerobic environment
Table 6 isoliquiritigenin is in the influence (uM concentration) to LDH-A of normal oxygen and anaerobic environment
80% eluate and the isoliquiritigenin activity to LDH-A under normal oxygen and anoxia condition all has the obvious suppression effect, and has the relation that tangible dose-effect relies on.The effect of isoliquiritigenin is better than the effect of 80% eluate.
3,80% pure eluate and isoliquiritigenin are to reaching the influence to its apoptosis at body tumor tissues LDH-A protein expression
Adopt the method for tumor cell transplantation to set up breast carcinoma nude mice animal model, at the position of mammary gland injection 3*10
6, give 80% pure eluate dosage 60mg/kg, isoliquiritigenin dosage 20,40mg/kg; Inferior on every Wendesdays, successive administration 26 days is got tumor tissues; Measure the expression of LDH-A in each tissue with the method for SABC, and medicine is to the influence of tumor tissues apoptosis, the result sees Fig. 3,4.
The result shows that 80% ethanol extract and isoliquiritigenin can suppress the proteic expression of LDH-A in the tumor tissues; The LDH-A protein expression that matched group is found in the inspection of LDH-A tissue staining is apparently higher than the administration group; Explain that administration (SS or ISO) can both obviously suppress the expression of LDH-A in the tumor tissues, painted obvious reduction (P＜0.05) in two groups of different cells transplantation tumor tissues.
80% ethanol extract and isoliquiritigenin ability inducing apoptosis of tumour cell are with the level of apoptosis of cell in TUNEL (the original position enzyme labelling assay for determining of apoptosis) the method detection transplanted tumor in nude mice tissue.This dyeing brown yellow granule occurs in the nucleus of apoptotic cell; The administration group comprises SS; And the positive cell number of ISO group apoptosis obviously increases; The positive cell number of comparing apoptosis in the transplanted tumor tissue with the normal control group obviously increases (P＜0.05), is combined into tumor size, the whole weight of tumor body and final volume and obviously reduces (P＜0.05).
Conclusion: SS and ISO can obviously suppress the growth of transplanted tumor in MCF-7 and the MDA-MB-231 cell nude mouse; Its mechanism of action possibly be the expression through downward modulation LDH-A; And then reduce tumor cell energy for growth in vivo, have the effect that suppresses the breast tumor cell growth in the body.
Experimental example 3 the present invention carry out composition of prescription to isolating monomeric compound and observe its inhibitory action to LDH-A
1.1 experimental drug: isoliquiritigenin (ISO purchases in Nat'l Pharmaceutical & Biological Products Control Institute), nutgall catechin (GC), epicatechin (EC) and catechin (C) (all purchase in last Hiroad standing grain medical sci-tech Development Co., Ltd)
1.2 experimental technique: drug component is: the blank group; The ISO group; The GC group; The EC group; The C group; ISO+GC organizes (ISO: GC=5: 10); ISO+GC+EC organizes (ISO: GC: EC=5: 10: 200); And ISO+GC+EC+C organizes (5: 10: 200: 200).Take the logarithm the respectively MCF-7 of trophophase, the every hole 100 μ L of MDA-MB-231 are inoculated in 96 orifice plates, and cell concentration is 5 * 10
4/ mL; After cultivation 24h treats cell attachment under condition of normal pressure, change serum-free medium, absorb old culture medium behind the 24h; Add 90 μ L serum-free mediums more respectively and add above-mentioned medicine respectively; The blank group adds the phosphate buffer that 10 μ L contain 2 ‰ DMSO, establishes 6 parallel holes for every group, places 37 ℃, 5%CO
2Cultivate 48h in the cell culture incubator.Behind the collecting cell, extract total protein, measure the influence of medicine pair cell LDH-A protein expression with Western blot (β-actin is confidential reference items).
1.3 experimental result: result such as table 7, Fig. 5.
Table 7 C, EC, GC, ISO are separately to the influence of LDH-A
| Matched group | C | EC | GC | ISO | |
| MDA-MB-231 | 0.9±0.06 | 0.8±0.06 | 0.8±0.05 | 0.8±0.05 | 0.35±0.04 |
| MCF-7 | 0.8±0.04 | 0.8±0.04 | 0.78±0.03 | 0.76±0.03 | 0.56±0.04 |
Can find out that by table 7 ISO shows inhibitory action to the expression of LDH-A separately; GC, EC, C all do not have the obvious suppression effect to the expression of LDH-A separately; Can find out that by Fig. 5 GC, EC, C three unite use, to the expression unrestraint effect of LDH-A; And as ISO and GC, EC, obviously strengthen inhibitory action (protein band is obviously thin out, and promptly protein expression is suppressed) when the C compatibility uses to LDH-A.This shows that the relevant monomer compatibility of ISO and Caulis Spatholobi can obviously strengthen the inhibitory action to LDH-A, thereby increase the effect that it suppresses tumor.
Description of drawings:
Fig. 1: the influence that 80% eluate is expressed two types of different tumor cell globin matter.
Fig. 2: the influence that isoliquiritigenin is expressed two types of different tumor cell globin matter.
Fig. 3: 80% eluate is to the influence of tumor tissues LDH-A and apoptosis.
Fig. 4: isoliquiritigenin is to the influence of tumor tissues LDH-A and apoptosis.
Fig. 5: the monomeric compound compatibility uses the active influence of tumor LDH-A.
The specific embodiment
Embodiment 1
Caulis Spatholobi medicinal material coarse powder 100g decoct to extract 2 times with 10 times of water (W/V), each 1 hour, merge extracted twice liquid, be evaporated to dried, Caulis Spatholobi water extract 9.7g.Water extract adds conventional adjuvant and processes granule according to common process.
Embodiment 2
Caulis Spatholobi medicinal material coarse powder 100g, with 60% alcohol reflux of 10 times of amounts (W/V) times 2 times, each 1 hour, merge extracted twice liquid, be evaporated to dried, Caulis Spatholobi 60% ethanol extract 15.5g.Ethanol extract adds conventional adjuvant and processes drop pill according to common process.
Embodiment 3
Caulis Spatholobi medicinal material coarse powder 1000g; With 60% ethanol ultrasonic extraction of 10 times of amounts (W/V) 2 times; Each 1 hour, merge extracted twice liquid, be evaporated to do not have the alcohol flavor after; Extract successively with petroleum ether, ethyl acetate, n-butyl alcohol respectively, obtain 4 extracts that polarity is different: petroleum ether extract (PE) 1.1g, acetic acid ethyl ester extract (ETOAC) 11.5g, n-butyl alcohol extract-I (BUOH-I) 24.4g and water layer stay excess (DW) 79.3g.
Embodiment 4
The Caulis Spatholobi medicinal material coarse powder, with 60% ethanol ultrasonic extraction of 10 times of amounts (W/V) 2 times, each 1 hour; Merge extracted twice liquid, be evaporated to do not have the alcohol flavor after, with an amount of n-butanol extraction 3 times; Combining extraction liquid, be concentrated into dried, n-butyl alcohol extract-II (BuOH-II).Get BuOH-II, behind the water dispersing and dissolving, last D101 macroporous adsorptive resins adsorbs.Elder generation's water be washed till effluent colourless after, successively with 20%, 40% ethanol water, 3 times of column volumes of eluting respectively, use 3 times of column volumes of 80% ethanol elution at last again, eluent is evaporated to dried, must 80% ethanol elution thing.
Embodiment 5
The Caulis Spatholobi medicinal material coarse powder, with 60% ethanol ultrasonic extraction of 10 times of amounts (W/V) 2 times, each 1 hour; Merge extracted twice liquid, be evaporated to do not have the alcohol flavor after, with an amount of n-butanol extraction 3 times; Combining extraction liquid, be concentrated into dried, n-butyl alcohol extract-II (BuOH-II).Get BuOH-II, behind the water dispersing and dissolving, last D101 macroporous adsorptive resins adsorbs.Elder generation's water be washed till effluent colourless after, successively with 20%, 40% ethanol water, 3 times of column volumes of eluting respectively, use 3 times of column volumes of 60% ethanol elution at last again, eluent is evaporated to dried, 60% ethanol elution thing must extract.
According to the method experiment of experimental example 2, the result shows that this extract has inhibitory action to the expression of LDH-A, can inducing apoptosis of tumour cell.
Embodiment 6
The Caulis Spatholobi medicinal material coarse powder, with 60% ethanol ultrasonic extraction of 10 times of amounts (W/V) 2 times, each 1 hour; Merge extracted twice liquid, be evaporated to do not have the alcohol flavor after, with an amount of n-butanol extraction 3 times; Combining extraction liquid, be concentrated into dried, n-butyl alcohol extract-II (BuOH-II).Get BuOH-II, behind the water dispersing and dissolving, last D101 macroporous adsorptive resins adsorbs.Elder generation's water be washed till effluent colourless after, successively with 20%, 40% ethanol water, 3 times of column volumes of eluting respectively, use 3 times of column volumes of 90% ethanol elution at last again, eluent is evaporated to dried, 90% ethanol elution thing must extract.
According to the method experiment of experimental example 2, the result shows that this extract has inhibitory action to the expression of LDH-A, can inducing apoptosis of tumour cell.
The Caulis Spatholobi medicinal material coarse powder, with 60% ethanol ultrasonic extraction of 10 times of amounts (W/V) 2 times, each 1 hour; Merge extracted twice liquid, be evaporated to do not have the alcohol flavor after, with an amount of n-butanol extraction 3 times; Combining extraction liquid, be concentrated into dried, n-butyl alcohol extract-II (BUOH-II).Get BUOH-II, behind the water dispersing and dissolving, last D101 macroporous adsorptive resins adsorbs.Elder generation's water be washed till effluent colourless after, successively with 20%, 40% ethanol water, 3 times of column volumes of eluting respectively, use 3 times of column volumes of 80% ethanol elution at last again, eluent is evaporated to dried, must 80% ethanol elution thing.
Embodiment 8
Get isoliquiritigenin (ISO) and nutgall catechin (GC), epicatechin (EC) and catechin (C) in quality ISO: GC: EC: C=5: 10: 200: 200 ratios totally 100 grams mix monomer composition.
Embodiment 9
Get isoliquiritigenin (ISO) and nutgall catechin (GC), epicatechin (EC) and catechin (C) in quality ISO: GC: EC: C=2: 14: 160: 240 ratios totally 100 grams mix monomer composition.
Embodiment 10
Get isoliquiritigenin (ISO) and nutgall catechin (GC), epicatechin (EC) and catechin (C) in quality ISO: GC: EC: C=9: 6: 240: 140 ratios totally 100 grams mix monomer composition.
Embodiment 11
The Caulis Spatholobi medicinal material coarse powder, with 60% ethanol ultrasonic extraction of 10 times of amounts (W/V) 2 times, each 1 hour; Merge extracted twice liquid, be evaporated to do not have the alcohol flavor after, with an amount of n-butanol extraction 3 times; Combining extraction liquid, be concentrated into dried, n-butyl alcohol extract-II (BUOH-II).Get 3UOH-II, behind the water dispersing and dissolving, last D101 macroporous adsorptive resins adsorbs.Elder generation's water be washed till effluent colourless after, distinguish 3 times of column volumes of eluting with 20%, 40% ethanol water successively again, use 3 times of column volumes of 80% ethanol elution at last, eluent is evaporated to dried, 80% ethanol elution thing.
Getting 80% eluate adds conventional adjuvant and processes nanometer formulation according to common process.
Embodiment 12
Caulis Spatholobi medicinal material coarse powder 200g, with 60% ethanol ultrasonic extraction of 10 times of amounts (W/V) 2 times, each 1 hour; Merge extracted twice liquid, be evaporated to after do not have the alcohol flavor, with an amount of n-butanol extraction 3 times; Combining extraction liquid, be concentrated into dried, n-butyl alcohol extract-II (BUOH-II) 38.2g.Take by weighing BUOH-II5g, behind the water dispersing and dissolving, last D101 macroporous adsorptive resins adsorbs.The elder generation water be washed till effluent colourless after; Again successively with 3 times of column volumes of 20%, 40%, 80% ethanol water difference eluting; Collect each eluting stream part; Be evaporated to dried, 20% ethanol elution thing (Fr.20%) 1.12,40% ethanol elution thing (Fr.40%) 0.56g and 80% ethanol elution thing (Fr.80%) 2.33g.
Gained Fr.80%102mg separates through silica gel column chromatography, ODS preparative high-performance liquid chromatographic, obtains chemical compound isoliquiritigenin (2.8mg).
Isoliquiritigenin (isoliquiritigenin, ISO)
ESI-MS：m/z?255[M-H]
-，
UV λ max:205nm, 239nm (acromion), 368.5nm.
1H-NMR(400MHz，DMSO-d
6)：δ13.51(1H，2’-OH)，10.56(1H，4’-OH)，10.08(1H，4-OH)，8.15(1H，d，J＝8.7Hz，H-6’)，7.77(1H，d，J＝15.8Hz，H-)，7.75(2H，d，J＝8.0Hz，H-2，6)，7.72(1H，d，J＝15.8Hz，H-)，6.82(2H，d，J＝8.0Hz，H-3，5)，6.38(1H，dd，J＝8.7，1.5Hz，H-5’)，6.25(1H，d，J＝1.5Hz，H-3’)。
13C-NMR(100MHz，DMSO-d6)：δ126.0(C-1)，130.6(C-2)，116.1(C-3)，160.1(C-4)，116.0(C-5)，130.6(C-6)，144.2(C-)，117.8(C-)，191.7(C＝O)，113.8(C-1’)，164.6(C-2’)，102.5(C-3’)，165.6(C-4’)，107.5(C-5’)，132.1(C-6’)。
Embodiment 13
The Caulis Spatholobi medicinal material coarse powder, with 50% ethanol percolate extraction of 10 times of amounts (W/V) 2 times, each 1 hour; Merge extracted twice liquid, be evaporated to do not have the alcohol flavor after, with an amount of n-butanol extraction 3 times; Combining extraction liquid, be concentrated into dried, n-butyl alcohol extract-II (BuOH-II).Get BuOH-II, behind the water dispersing and dissolving, last D101 macroporous adsorptive resins adsorbs.Elder generation's water be washed till effluent colourless after, distinguish 3 times of column volumes of eluting with 20%, 40% ethanol water successively again, use 3 times of column volumes of 80% ethanol elution at last, eluent is evaporated to dried, 80% ethanol elution thing;
According to the experiment of the method for experimental example 2, the result shows that this extract can suppress the growth of tumor cell, and the expression of LDH-A is had inhibitory action, can inducing apoptosis of tumour cell.
Embodiment 14
The Caulis Spatholobi medicinal material coarse powder is extracted 2 times with 10 times of water gagings, each 1 hour, merge extracted twice liquid, with an amount of n-butanol extraction 3 times, combining extraction liquid, be concentrated into dried, must n-butyl alcohol extract-II (BUOH-II).Get BUOH-II, behind the water dispersing and dissolving, last D101 macroporous adsorptive resins adsorbs.Elder generation's water be washed till effluent colourless after, distinguish 3 times of column volumes of eluting with 20%, 40% ethanol water successively again, use 3 times of column volumes of 80% ethanol elution at last, eluent is evaporated to dried, 80% ethanol elution thing;
According to the experiment of the method for experimental example 2, the result shows that this extract can suppress the growth of tumor cell, and the expression of LDH-A is had inhibitory action, can inducing apoptosis of tumour cell.
Embodiment 15
The Caulis Spatholobi medicinal material coarse powder, with 70% ethanol percolate extraction of 10 times of amounts (W/V) 2 times, each 1 hour; Merge extracted twice liquid, be evaporated to do not have the alcohol flavor after, with an amount of n-butanol extraction 3 times; Combining extraction liquid, be concentrated into dried, n-butyl alcohol extract-II (BUOH-II).Get BUOH-II, behind the water dispersing and dissolving, last D101 macroporous adsorptive resins adsorbs.Elder generation's water be washed till effluent colourless after, distinguish 3 times of column volumes of eluting with 20%, 40% ethanol water successively again, use 3 times of column volumes of 80% ethanol elution at last, eluent is evaporated to dried, 80% ethanol elution thing;
According to the experiment of the method for experimental example 2, the result shows that this extract can suppress the growth of tumor cell, and the expression of LDH-A is had inhibitory action, can inducing apoptosis of tumour cell.
Embodiment 16
Get isoliquiritigenin (ISO) and nutgall catechin (GC), epicatechin (EC) by ISO: GC: EC=5: mixed got monomer composition in 10: 200: 200.
Embodiment 17
Get isoliquiritigenin (ISO) and nutgall catechin (GC) in ISO: GC=2: 14 ratios totally 100 grams mix monomer composition.
Embodiment 18
Get isoliquiritigenin (ISO), epicatechin (EC) by ISO: EC=9: 240 mass ratios totally 100 grams mix monomer composition.
Claims (10)
1. isoliquiritigenin prevents or treats the application in the three negative breast cancer medicines in preparation.
2. the Caulis Spatholobi extract that contains isoliquiritigenin is preparing prevention or treating the application in the three negative breast cancer medicines, and said Caulis Spatholobi extract is: Caulis Spatholobi water extract, ethanol extraction or Caulis Spatholobi water extract, ethanol extraction successively extract or select the extract that an extraction obtains successively with petroleum ether, ethyl acetate, n-butyl alcohol.
3. application as claimed in claim 2; It is characterized in that described Caulis Spatholobi extract is prepared by following method: use n-butanol extraction behind Caulis Spatholobi water extraction or the ethanol extraction, extract is macroporous resin on conventional method, uses earlier water elution; The ethanol liquid eluting that the back increases with concentration in 10-95% concentration of alcohol scope successively gradually; Comprising using the 60-90% ethanol elution, collect 60-90% ethanol elution thing, promptly get.
4. application as claimed in claim 3 is characterized in that said macroporous resin is a low pole.
5. like each described application of claim 2-4, it is characterized in that ethanol extraction uses the 10-95% ethanol extraction.
6. application as claimed in claim 5 is characterized in that the ethanol extraction mode is percolation, backflow, dipping or supersound extraction.
7. Caulis Spatholobi extract that contains isoliquiritigenin; It is characterized in that this extract is prepared by following method: use n-butanol extraction behind Caulis Spatholobi water extraction or the ethanol extraction, extract is macroporous resin on conventional method, uses earlier water elution; The ethanol liquid eluting that the back increases with concentration in 10-95% concentration of alcohol scope successively gradually; Comprising using the 60-90% ethanol elution, collect 60-90% ethanol elution thing, promptly get.
8. a pharmaceutical composition that prevents or treat three negative breast cancer is characterized in that the active component of said composition mainly is made up of one or more active component in isoliquiritigenin and optional nutgall catechin, epicatechin and the catechin.
9. compositions as claimed in claim 8 is characterized in that said composition reaches one or more active component of choosing wantonly in nutgall catechin 5-15 weight portion, epicatechin 150-250 weight portion, the catechin 150-250 weight portion by isoliquiritigenin 1-9 weight portion and forms.
10. the pharmaceutical composition like claim 8 or 9 prevents or treats the application in the three negative breast cancer medicines in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210019695.9A CN102579425B (en) | 2012-01-16 | 2012-01-16 | Caulis Spatholobi extract, application thereof and new application of isoliquiritigenin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210019695.9A CN102579425B (en) | 2012-01-16 | 2012-01-16 | Caulis Spatholobi extract, application thereof and new application of isoliquiritigenin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102579425A true CN102579425A (en) | 2012-07-18 |
| CN102579425B CN102579425B (en) | 2014-07-09 |
Family
ID=46469036
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210019695.9A Active CN102579425B (en) | 2012-01-16 | 2012-01-16 | Caulis Spatholobi extract, application thereof and new application of isoliquiritigenin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102579425B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106138186A (en) * | 2015-04-16 | 2016-11-23 | 陈晓萍 | A kind of prevention or the external preparation for the treatment of breast cancer |
| CN108685998A (en) * | 2018-08-02 | 2018-10-23 | 广西壮族自治区药用植物园 | Treat the preparation method of breast cancer Caulis Spatholobi tablet |
| CN108969731A (en) * | 2018-10-12 | 2018-12-11 | 广西壮族自治区药用植物园 | The preparation method for treating the Caulis Spatholobi ointment of breast cancer |
| CN109200086A (en) * | 2018-10-12 | 2019-01-15 | 广西壮族自治区药用植物园 | The preparation method for treating the Caulis Spatholobi tablet of oophoroma |
| CN114099528A (en) * | 2021-11-25 | 2022-03-01 | 广西壮族自治区中医药研究院 | Caulis spatholobi antidepressant quality marker and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1939396A (en) * | 2006-10-26 | 2007-04-04 | 首都医科大学附属北京中医医院 | Suberect spatholobus stem extract with antineoplastic function and its making method |
| CN101658513A (en) * | 2008-08-26 | 2010-03-03 | 石河子大学 | Use of isoliquiritigenin as medicament for preventing and treating or curing postoperative metastasis and relapse of malignant tumors |
-
2012
- 2012-01-16 CN CN201210019695.9A patent/CN102579425B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1939396A (en) * | 2006-10-26 | 2007-04-04 | 首都医科大学附属北京中医医院 | Suberect spatholobus stem extract with antineoplastic function and its making method |
| CN101658513A (en) * | 2008-08-26 | 2010-03-03 | 石河子大学 | Use of isoliquiritigenin as medicament for preventing and treating or curing postoperative metastasis and relapse of malignant tumors |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106138186A (en) * | 2015-04-16 | 2016-11-23 | 陈晓萍 | A kind of prevention or the external preparation for the treatment of breast cancer |
| CN108685998A (en) * | 2018-08-02 | 2018-10-23 | 广西壮族自治区药用植物园 | Treat the preparation method of breast cancer Caulis Spatholobi tablet |
| CN108969731A (en) * | 2018-10-12 | 2018-12-11 | 广西壮族自治区药用植物园 | The preparation method for treating the Caulis Spatholobi ointment of breast cancer |
| CN109200086A (en) * | 2018-10-12 | 2019-01-15 | 广西壮族自治区药用植物园 | The preparation method for treating the Caulis Spatholobi tablet of oophoroma |
| CN114099528A (en) * | 2021-11-25 | 2022-03-01 | 广西壮族自治区中医药研究院 | Caulis spatholobi antidepressant quality marker and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102579425B (en) | 2014-07-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ryu et al. | Astragali Radix elicits anti-inflammation via activation of MKP-1, concomitant with attenuation of p38 and Erk | |
| Kim et al. | The inhibition of JNK MAPK and NF-κB signaling by tenuifoliside A isolated from Polygala tenuifolia in lipopolysaccharide-induced macrophages is associated with its anti-inflammatory effect | |
| Ravoori et al. | Inhibition of estrogen-mediated mammary tumorigenesis by blueberry and black raspberry | |
| Jiang et al. | Total Flavonoids from Radix Glycyrrhiza Exert Anti‐Inflammatory and Antitumorigenic Effects by Inactivating iNOS Signaling Pathways | |
| CN102579425B (en) | Caulis Spatholobi extract, application thereof and new application of isoliquiritigenin | |
| Liu et al. | Rosmarinic acid inhibits cell proliferation, migration, and invasion and induces apoptosis in human glioma cells | |
| CN105362229A (en) | Water-soluble phloretin solid dispersion and preparation method thereof | |
| Liu et al. | Icariin as an emerging candidate drug for anticancer treatment: Current status and perspective | |
| Kong et al. | Antitumor activity of enzymatically hydrolyzed Ganoderma lucidum polysaccharide on U14 cervical carcinoma–bearing mice | |
| Yue et al. | Effectiveness of Scutellaria barbata water extract on inhibiting colon tumor growth and metastasis in tumor‐bearing mice | |
| Lee et al. | A network pharmacology study on the molecular mechanisms of FDY003 for breast cancer treatment | |
| Zheng et al. | Ferulic acid enhances the chemical and biological properties of astragali radix: a stimulator for danggui buxue tang, an ancient Chinese herbal decoction | |
| Wang et al. | Molecular mechanism and pharmacokinetics of flavonoids in the treatment of resistant EGF receptor‐mutated non‐small‐cell lung cancer: A narrative review | |
| Hwang et al. | Anti-inflammatory effect of immature sword bean pod (Canavalia gladiata) in lipopolysaccharide-induced RAW264. 7 cells | |
| Shao et al. | Anti-inflammatory effects and molecular mechanisms of bioactive small molecule garlic polysaccharide | |
| Song et al. | Myricetin exhibit selective anti-lymphoma activity by targeting BTK and is effective via oral administration in vivo | |
| CN103505454A (en) | Alpha-tubulin inhibitor and application thereof | |
| Duan et al. | Trametenolic Acid Ameliorates the Progression of Diabetic Nephropathy in db/db Mice via Nrf2/HO‐1 and NF‐κB‐Mediated Pathways | |
| Luong Huynh et al. | Pharmacological properties of ginsenosides in inflammation-derived cancers | |
| Yang et al. | Combination effects of polyphenols present in sugarcane on proliferation in MCF-7 human breast cancer cells | |
| KR102271713B1 (en) | Pharmaceutical composition for preventing or treating cancer, comprising a Disocactus flagelliformis extract | |
| CN107056869B (en) | A kind of withanolide class compound and extracting method and application | |
| CN107583054A (en) | Cryptotanshinone pharmaceutical composition and its application in treatment chronic myelocytic leukemia medicine is prepared | |
| CN105213366A (en) | The medical usage of guttiferone compound and pharmaceutical composition thereof | |
| CN1919339B (en) | Cucurbitacin nano preparation comprising protein, preparation method and use thereof |
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 |