CN105327071A - Antineoplastic traditional Chinese medicinal composition and application thereof - Google Patents
Antineoplastic traditional Chinese medicinal composition and application thereof Download PDFInfo
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- CN105327071A CN105327071A CN201410384095.1A CN201410384095A CN105327071A CN 105327071 A CN105327071 A CN 105327071A CN 201410384095 A CN201410384095 A CN 201410384095A CN 105327071 A CN105327071 A CN 105327071A
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Abstract
The invention relates to the field of medicine, and in particular relates to an antineoplastic traditional Chinese medicinal composition. The traditional Chinese medicinal composition is prepared from the following medicinal raw materials in percentage by weight: 10-70% of airpotato yam rhizome and 30-90% of sculellaria barbata; and the traditional Chinese medicinal composition is a whole-powder suspension of the airpotato yam rhizome and the sculellaria barbata, or the traditional Chinese medicinal composition is an ethanol extract with concentration being 20-80%, an ethyl acetate extract, an acetone extract or a gasoline extract. The invention also relates to application of a sculellaria barbata extract and a traditional Chinese medicinal composition of the sculellaria barbata extract, wherein the traditional Chinese medicinal composition consists of the following ingredients: an ethanol extract of airpotato yam rhizome with concentration being 20-80% and an ethanol extract of sculellaria barbata with concentration being 20-80% at a weight ratio of (1-2) to (2-1). The invention also relates to application of scutellarin and a pharmaceutical composition of the scutellarin, wherein the traditional Chinese medicinal composition consists of the following ingredients: diosbulbin B and scutellarin at a weight ratio of (1-1) to (1-2). The pharmaceutical composition can be used for preventing and treating various cancers such as pancreatic cancer, breast cancer, lung cancer, colon cancer and the like.
Description
Technical field
The present invention relates to field of medicaments, particularly relate to a kind of antineoplastic Chinese medicine composition and medical usage thereof.
Background technology
Cancer is one of principal disease of serious threat human health and life.In treatment of cancer, although traditional Radiotherapy chemotherapy makes great progress, but because it is while killing and wounding cancerous cell, also to human body generation toxic and side effects in various degree, wherein mainly bone marrow depression and immunologic hypofunction, can leukopenia etc. be caused, therapy discontinued of sometimes even having to, thus also limit the curative effect of medicine.In order to overcome a series of disadvantages that traditional Radiotherapy chemotherapy brings, pharmacy worker constantly explores in recent years, and appliable plant medicine carries out the hot fields that anti-curing cancers becomes current research
Rhizoma Dioscoreae Bulbiferae is the dry tuber of Dioscoreaceae yam Dioscorea bulbifera L. DioscoreabulbiferaL., widely distributed in the subtropical and tropical zones in Asia, is also present in the some areas in Africa, and tradition it can be used as medicinal main in China.As far back as 500-600 in Christian era, Rhizoma Dioscoreae Bulbiferae powder, decoction or tincture, just be used to the treatment of the disease such as goiter (tumor), gastric cancer, rectal cancer, laryngalgia, pharyngitis, metrorrhagia in such as modern medicine, and wherein especially conventional with the treatment of goiter (tumor) and many types of cancer; On modern clinic, normal treatment Rhizoma Dioscoreae Bulbiferae being used for the many types of cancers such as the esophageal carcinoma, gastric cancer, hepatocarcinoma, rectal cancer, breast carcinoma, pulmonary carcinoma, cervical cancer, nasopharyngeal carcinoma, carcinoma of maxillary sinus, bladder cancer, is widely used, determined curative effect.But Rhizoma Dioscoreae Bulbiferae causes toxic hepatitis even dead in Clinical practice to have report to find, thus greatly limit the performance of its clinical good effects.
Compatibility is one of characteristic of middle medical drugs, can be played the effect of removing toxic substances potentiation by compatibility.Herba Scutellariae Barbatae is the dry herb of Labiatae Scutellaria plant Herba Scutellariae Barbatae ScutellariabarbataD.Don, originates in South America, is mainly distributed in the ground such as Argentina, South Brazil, Uruguay and China's Mainland.Its acrid in the mouth, hardship, cold in nature, there is effect of heat-clearing and toxic substances removing, blood stasis dispelling diuresis, be mainly used in treatment furuncle swelling toxin, laryngopharynx swelling and pain clinically, fall and flutter the pain of injury, edema, jaundice and worm venom.Chemical composition isolation identification shows, and the main chemical compositions of Herba Scutellariae Barbatae is flavone compound, also containing compositions such as diterpene-kind compound, polysaccharide, alkaloid and steroidals.Wherein scutellarin is the compound that in flavone compound, content is maximum, is also the effective ingredient of Herba Scutellariae Barbatae.Modern pharmacology research shows, Herba Scutellariae Barbatae has antibacterial, antitumor, protects the liver and anti-hepatic fibrosis isoreactivity.
Summary of the invention
Object of the present invention aims to provide a kind of antineoplastic Chinese medicine composition and uses thereof.
Specifically, a first aspect of the present invention there is provided a kind of antineoplastic Chinese medicine composition, it is characterized in that, described Chinese medicine composition is made up of the crude drug of following weight percents: Rhizoma Dioscoreae Bulbiferae 10% ~ 70%, Herba Scutellariae Barbatae 30% ~ 90%, and described Chinese medicine composition is the full powder suspension of Rhizoma Dioscoreae Bulbiferae and Herba Scutellariae Barbatae, or its 20% ~ 80% ethanol extraction, ethyl acetate extract, acetone extract or ligroin extraction.
In a preference, the Rhizoma Dioscoreae Bulbiferae composition in described Chinese medicine composition is containing diosbulbin B 1% ~ 17%, and the Herba Scutellariae Barbatae composition in described Chinese medicine composition is containing scutellarin 1% ~ 15%.
A second aspect of the present invention there is provided the application of above-mentioned Chinese medicine composition in the medicine or health product of preparation control cancer of pancreas, breast carcinoma, pulmonary carcinoma, gastric cancer, hepatocarcinoma, bladder cancer, ovarian cancer, colon cancer, carcinoma of prostate, the esophageal carcinoma, skin carcinoma, thyroid carcinoma, leukemia or cervical cancer.
A third aspect of the present invention there is provided a kind of purposes of Herba Scutellariae Barbatae extract, described Herba Scutellariae Barbatae extract can be used for the anti-tumor activity improving Rhizoma Dioscoreae Bulbiferae extract, and described Herba Scutellariae Barbatae extract and Rhizoma Dioscoreae Bulbiferae extract are respectively Herba Scutellariae Barbatae 20% ~ 80% ethanol extraction and Rhizoma Dioscoreae Bulbiferae 20% ~ 80% ethanol extraction.
In a preference, the weight ratio of described Herba Scutellariae Barbatae extract and Rhizoma Dioscoreae Bulbiferae extract is 1:2 ~ 2:1.
In another preference, described Herba Scutellariae Barbatae extract is containing scutellarin 1% ~ 6%, and described Rhizoma Dioscoreae Bulbiferae extract is containing diosbulbin B 1% ~ 7%.
A third aspect of the present invention there is provided a kind of antineoplastic Chinese medicine composition, and described Chinese medicine composition is grouped into by the one-tenth of following weight ratio: Rhizoma Dioscoreae Bulbiferae 20% ~ 80% ethanol extraction: Herba Scutellariae Barbatae 20% ~ 80% ethanol extraction=1:2 ~ 2:1.
In a preference, described Rhizoma Dioscoreae Bulbiferae extract is containing diosbulbin B 1% ~ 7%, and described Herba Scutellariae Barbatae extract is containing scutellarin 1% ~ 6%.
A fourth aspect of the present invention there is provided the application of above-mentioned Chinese medicine composition in the medicine or health product of preparation control cancer of pancreas, breast carcinoma, pulmonary carcinoma, gastric cancer, hepatocarcinoma, bladder cancer, ovarian cancer, colon cancer, carcinoma of prostate, the esophageal carcinoma, skin carcinoma, thyroid carcinoma, leukemia or cervical cancer.
A fifth aspect of the present invention there is provided a kind of purposes of scutellarin, and described scutellarin can be used for the anti-tumor activity improving diosbulbin B
In a preference, the weight ratio of described scutellarin and diosbulbin B is 1:1 ~ 2:1.
A sixth aspect of the present invention there is provided a kind of antitumor medicine composition, and described Chinese medicine composition is grouped into by the one-tenth of following weight ratio: diosbulbin B: scutellarin=1:1 ~ 1:2.
A seventh aspect of the present invention there is provided the application of aforementioned pharmaceutical compositions in the medicine or health product of preparation control cancer of pancreas, breast carcinoma, pulmonary carcinoma, gastric cancer, hepatocarcinoma, bladder cancer, ovarian cancer, colon cancer, carcinoma of prostate, the esophageal carcinoma, skin carcinoma, thyroid carcinoma, leukemia or cervical cancer.
The details of various aspects of the present invention is able to detailed description by chapters and sections subsequently.By hereafter and the description of claim, feature of the present invention, object and advantage will be more obvious.
Detailed description of the invention
Appearance part of the present invention is based on so unexpected discovery: the combination of Rhizoma Dioscoreae Bulbiferae and Herba Scutellariae Barbatae or its extract significantly can suppress the growth of kinds of tumors.Therefore, above-mentioned composition can be expected to develop the medicine becoming a kind of prevention or treatment tumor.And the combination of the Rhizoma Dioscoreae Bulbiferae of specific proportioning and the extract of Herba Scutellariae Barbatae or its active component has the beyond thought effect of Synergistic antineoplastic especially.
The preparation process that the present invention has the Chinese medicine composition of anti-curing cancers effect there is no special feature.Rhizoma Dioscoreae Bulbiferae tuber and Herba Scutellariae Barbatae can be beaten powder respectively, by reflux, extract, such as water, 20% ~ 80% ethanol, ethyl acetate, acetone or petroleum ether, add common medicinal adjuvant and make any one pharmaceutically said dosage form, preferred extractum, fluid extract, medicated wine, tincture, oral liquid, pill, powder, drop pill, tablet or capsule.
It is closely related that Chinese medicine composition of the present invention has the synergism of anti-curing cancers effect probably and between following material: the diterpene lactone material in Rhizoma Dioscoreae Bulbiferae, especially the flavones ingredient in diosbulbin B and Herba Scutellariae Barbatae, the scutellarin especially in Herba Scutellariae Barbatae.The structural formula of diosbulbin B (DiosbulbinB, DB) and scutellarin (Scutellarin, SC) is as follows:
Effect experiment subsequently will prove, the kinds cancers such as Chinese medicine composition of the present invention alternative chemotherapeutic control cancer of pancreas, breast carcinoma, pulmonary carcinoma, gastric cancer, hepatocarcinoma, bladder cancer, ovarian cancer, colon cancer, carcinoma of prostate, the esophageal carcinoma, skin carcinoma, thyroid carcinoma, leukemia or cervical cancer.
Chinese medicine composition of the present invention is the plant extract of the pure natural of dietotherapeutic, and it is while high-efficiency prevention and control cancer, can also avoid using the toxic and side effects such as the hepatic injury that chemotherapeutic causes.
Chinese medicine composition of the present invention can be used alone or uses in the form of a pharmaceutical preparation.Pharmaceutical preparation comprises Chinese medicine composition of the present invention as active component and pharmaceutically suitable carrier.Preferably, Chinese medicine composition of the present invention contain 0.1-99.9% percentage by weight as the Rhizoma Dioscoreae Bulbiferae of the present invention of active component and the combination of Herba Scutellariae Barbatae or its extract or its active component." pharmaceutically suitable carrier " can not destroy the pharmaceutical active of Rhizoma Dioscoreae Bulbiferae of the present invention and Herba Scutellariae Barbatae or its extract or its active component, simultaneously its effective dose, and consumption when can play pharmaceutical carrier effect is to human non-toxic.
Described pharmaceutically suitable carrier includes but not limited to: soft phospholipid, aluminium stearate, aluminium oxide, ion exchange material, self-emulsifying drug delivery system, tween or other surfactants, serum albumin, buffer substance are if phosphate, glycine, sorbic acid, water, salt, electrolyte are as sulfate protamine, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, magnesium silicate, satisfied fatty acid partial glyceride mixtures etc.
Other conventional excipient substances are as binding agent (as microcrystalline Cellulose), filler (as starch, glucose, Lactis Anhydrous and lactose beadlet), disintegrating agent (as cross-linked pvp, crosslinked carboxymethyl fecula sodium, cross-linking sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose), lubricant (as magnesium stearate) and absorption enhancer, absorption carrier, flavouring agent, sweeting agent, excipient, diluent, wetting agent etc.
Chinese medicine composition of the present invention and pharmaceutical preparation thereof can by the preparations of this area conventional method and can by intestinal or non-bowel or topical routes.Oral formulations comprises capsule, tablet, oral liquid, granule, pill, powder, sublimed preparation, unguentum etc.; Non-intestinal drug delivery agent comprises injection etc.; Local administration preparation comprises cream, patch, ointment, spray etc.Be preferably oral formulations.
The route of administration of Chinese medicine composition of the present invention and its pharmaceutical preparation can be oral, Sublingual, percutaneous, through muscle or subcutaneous, mucocutaneous, vein, urethra, vagina etc.
Except making medicament, also can add the various food additive such as antioxidant, pigment, enzyme preparation in Chinese medicine composition of the present invention, make health food by the conventional method of this area.
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, the usually conveniently conditioned disjunction condition of advising according to manufacturer.Unless otherwise indicated, otherwise all percent, ratio, ratio or number by weight.
Unless otherwise defined, all specialties used in literary composition and scientific words and one skilled in the art the same meaning be familiar with.In addition, any method similar or impartial to described content and material all can be applicable in the inventive method.The use that better implementation method described in literary composition and material only present a demonstration.
The above-mentioned feature that the present invention mentions, or the feature that embodiment is mentioned can combination in any.All features that patent specification discloses can with any composition forms and use, each feature disclosed in description, anyly can provide identical, alternative characteristics that is impartial or similar object replaces.Therefore apart from special instruction, the feature disclosed is only general example that is impartial or similar features.
Embodiment 1 (impact of Rhizoma Dioscoreae Bulbiferae ethanol extract, Herba Scutellariae Barbatae ethanol extract, Rhizoma Dioscoreae Bulbiferae ethanol extract and Herba Scutellariae Barbatae ethanol extract Compatibility on Mice euphorbia egg decoctum solid tumor)
First the present invention investigates Rhizoma Dioscoreae Bulbiferae ethanol extraction (ET) with Herba Scutellariae Barbatae ethanol extraction (SE) compositions to the impact of S180 sarcoma solid tumor.
1.1 experiment material
1.1.1 animal subject
ICR mice, ♂, body weight 18 ~ 22g, purchased from Shanghai Slac Experimental Animal Co., Ltd., the animal quality certification number: SCXK (Shanghai 2007 – 0005).Mouse feeder temperature (22 ± 1) DEG C, relative humidity (65 ± 10) %, lighting hours 12h every day.
1.1.2 test medicine and preparation thereof
Rhizoma Dioscoreae Bulbiferae ethanol extraction (ET): take Rhizoma Dioscoreae Bulbiferae coarse powder 4000g, add 10 times of volume 80% alcohol reflux 3h, extract 4 times, merge extractive liquid, is evaporated to dry.Obtain Rhizoma Dioscoreae Bulbiferae ethanol extraction (ET).
Herba Scutellariae Barbatae ethanol extract (SE): weigh Herba Scutellariae Barbatae coarse powder 4000g, with 10 times of volume 80% alcohol reflux 2h after immersion 2h, extracts 3 times, merges 3 filtrates, concentrating under reduced pressure, dry, be Herba Scutellariae Barbatae ethanol extract (SE).
Rhizoma Dioscoreae Bulbiferae Herba Scutellariae Barbatae compositions: by aforementioned Rhizoma Dioscoreae Bulbiferae ethanol extraction and Herba Scutellariae Barbatae ethanol extract in ratio (weight ratio) combination of 2/1 ~ 1/2, obtain the compositions ET+SE of Rhizoma Dioscoreae Bulbiferae Herba Scutellariae Barbatae extract.
1.2 experimental technique
1.2.1 the content of diosbulbin B in high-performance liquid chromatogram determination EF
Chromatographic column is ultimateTMXB-C18column (4.6 × 250mm, 5 μm); Mobile phase condition is acetonitrile-water=25:75 (v/v), and flow velocity is 1.0mL/min, isocratic elution; Sampling volume is 10 μ L; Column temperature is 25 DEG C; Determined wavelength is 210nm.First, get diosbulbin B (DB) appropriate, accurately weighed, add the solution that methanol is made into 1mg/ml, mixing, obtains reference substance solution.DB (1000 μ g/mL) methanol dilution extremely a series of concentration: 7.81,15.63,31.25,62.5,125,250 μ g/mL, mixing, filters with 0.45 μm of organic facies filter membrane, get subsequent filtrate sample introduction, run according to above-mentioned liquid-phase condition, integration tries to achieve peak area value, and result is carried out regression analysis with concentration (μ g/mL) to peak area value and obtained standard curve and correlation coefficient.Secondly, get Rhizoma Dioscoreae Bulbiferae ethanol extraction (ET) appropriate, accurately weighed, put in 25ml volumetric flask, add 70% EtOH Sonicate and make it to dissolve, be settled to scale after cooling, filter with 0.45 μm of organic facies filter membrane.Get obtained need testing solution to run according to above-mentioned liquid-phase condition, the peak area value of gained substitutes into above-mentioned standard curve, calculates DB content.
1.2.2 the content of scutellarin in high-performance liquid chromatogram determination SE
Chromatographic column is SepaxHP-C18column (4.6 × 250mm, 5 μm); Mobile phase condition is methanol-water-acetic acid=35:61:4 (v/v/v), and flow velocity is 1.0mL/min, isocratic elution; Sampling volume is 5 μ L; Column temperature is 25 DEG C; Determined wavelength is 335nm.First get scutellarin (SC) appropriate, accurately weighed, add the solution that methanol is made into 1mg/ml, mixing, obtains reference substance solution.SC (1000 μ g/mL) methanol dilution extremely a series of concentration: 15.63,31.25,62.5,125,250,500,1000 μ g/mL, mixing, filters with 0.45 μm of organic facies filter membrane, gets subsequent filtrate sample introduction, run according to above-mentioned liquid-phase condition, integration tries to achieve peak area value, and result carries out regression analysis with concentration (μ g/mL) to peak area value, obtains standard curve and correlation coefficient.Secondly, get Herba Scutellariae Barbatae ethanol extract (SE) appropriate, accurately weighed, be placed in 25mL volumetric flask, add 80% ethanol to about 25mL, supersound extraction 60min, add 80% ethanol after returning to room temperature and be settled to 25mL, after mixing, filter with 0.45 μm of organic facies filter membrane.Get obtained need testing solution to run according to above-mentioned liquid-phase condition, the peak area value of gained substitutes into above-mentioned standard curve, calculates SC content.
1.2.3 tumor-bearing mice anti-tumor experiment in body
Adopt tumor-bearing mice experiment in vivo, in euphorbia egg decoctum tumor-bearing mice abdomen, extract corresponding tumor cell, with Trypan Blue, counting, is diluted to (1.0-1.3) × 10 with physiological saline solution
7cells/mL is in centrifuge tube, and insert on ice, subcutaneous injection (sc) inoculates this concentration tumor cell in the right oxter of ICR mice, and injection volume 0.1mL/ only.By body weight random packet, be respectively Control (CMC-Na), 5-Fu, Herba Scutellariae Barbatae ethanol extract SE, Rhizoma Dioscoreae Bulbiferae ethanol extraction ET, ET+SE group.Each extract or monomeric compound 0.5%CMC-Na are dissolved to desired concn, after inoculation, 24h starts by body weight gavage (ig) administration, CMC-Na group ig gives the 0.5%CMC-Na of equal volume, 5-Fu (with dd water dissolution to 2.5mg/mL) organizes lumbar injection (ip), every other day injects once; Successive administration 12d, 24h after last administration, pluck eyeball blood sampling, for measuring Serum ALT and AST vigor after centrifugalize serum; Get tumor tissue, weigh, take pictures.Press formulae discovery tumor control rate (Tumorinhibitionratio).
Tumorinhibitionratio (%)=[(C? T)/C] × 100, wherein C refers to the average tumor weight of CMC-Na matched group, and T shows medicine group average tumor weight.Heavily have significant difference with tumor, and tumor control rate >30% is the standard that medicine has anti-tumor activity.
1.3 statistical procedures
Experimental data all represents with meansigma methods ± standard error, analyzes with SPSS11.5 statistical software, carries out variance analysis in One-WayANOVA mode, and P<0.05 is for having statistical significant difference standard.
1.4 experimental result
1.4.1ET, SE and compositions ET+SE thereof is on the impact of S180 sarcoma solid tumor
Compare with model group, as shown in table 1, the mice of inoculation euphorbia egg decoctum is after giving ET (100mg/kg) 12d continuously, and tumor representation work reduces (P<0.05), shows that ET (100mg/kg) is effective to S180 solid tumor; Compare with ET (100mg/kg), compatibility SE (50mg/kg, 100mg/kg, 200mg/kg), tumor weight average significantly reduces (P<0.05), tumor control rate significantly improves, and shows that Herba Scutellariae Barbatae ethanol extract can strengthen the anti-tumor activity of Rhizoma Dioscoreae Bulbiferae ethanol extraction.
Table 1EF, SE and compositions ET+SE thereof is on the impact (n=10) of S180 sarcoma solid tumor
Compare with model group, * * * P<0.001; Compare with ET,
##p<0.01
1.4.2 on the impact of Biochemical Indices In Serum
As shown in Table 2, positive drug 5-Fu can make S180 tumor-bearing mice serum liver damage criterion ALT and AST all extremely significantly raise (P<0.01), show the liver toxicity of 5-Fu induction of tumor-bearing mice, and Chinese medicine composition of the present invention is while maintenance curative effect, hepatic injury index is had no significant effect, illustrates that Chinese medicine composition of the present invention has the advantage avoiding the side effect of chemotherapeutic 5-Fu induced liver injury.
Table 2 compositions EF+SE is on the impact of S180 sarcoma Biochemical Indices In Serum
Compare with model group, * * P<0.01
1.4.3 the content of diosbulbin B and scutellarin in Rhizoma Dioscoreae Bulbiferae and Herba Scutellariae Barbatae ethanol extract
Measure the content of diosbulbin B and scutellarin in Rhizoma Dioscoreae Bulbiferae and Herba Scutellariae Barbatae ethanol extract, find that in Rhizoma Dioscoreae Bulbiferae ethanol extract, diosbulbin B content is 6.42%, in Herba Scutellariae Barbatae ethanol extract, Content Measurement of Scutellarin is 4.09%.
Embodiment 2 (the antitumor action research of Rhizoma Dioscoreae Bulbiferae extract different from Herba Scutellariae Barbatae)
After determining Rhizoma Dioscoreae Bulbiferae Herba Scutellariae Barbatae compositions and having stronger anti-S180 solid tumor effect, the present invention has investigated the Anticancer effect in vivo of other extracts of Rhizoma Dioscoreae Bulbiferae and Herba Scutellariae Barbatae further.
2.1 experiment material
2.1.1 animal subject (1.1.1 with embodiment 1)
2.1.2 test medicine and preparation thereof
The full powder suspension of Rhizoma Dioscoreae Bulbiferae: Rhizoma Dioscoreae Bulbiferae 36g, pulverizes, and crosses 150 mesh sieves, is suspended in 180mL distilled water.
The full powder suspension of Herba Scutellariae Barbatae: Herba Scutellariae Barbatae 36g, pulverizes, and crosses 150 mesh sieves, is suspended in 180mL distilled water.
Rhizoma Dioscoreae Bulbiferae Aqueous extracts: Rhizoma Dioscoreae Bulbiferae 36g, adds 10 times of water gagings, reflux, extract, 3 times, each 3h, and filter merging filtrate, recycle-water makes into concentration 0.4g/mL.
Herba Scutellariae Barbatae Aqueous extracts: Herba Scutellariae Barbatae 36g, adds 10 times of water gagings, reflux, extract, 3 times, each 3h, and filter merging filtrate, recycle-water makes into concentration 0.4g/mL.
Rhizoma Dioscoreae Bulbiferae 70% ethanol extract: Rhizoma Dioscoreae Bulbiferae 36g, adds 10 times amount 70% ethanol, reflux, extract, 3 times, each 3h, filters merging filtrate, reclaims ethanol and makes into concentration 0.4g/mL.
Herba Scutellariae Barbatae 70% ethanol extract: Herba Scutellariae Barbatae 36g, adds 10 times amount 70% ethanol, reflux, extract, 3 times, each 3h, filters merging filtrate, reclaims ethanol and makes into concentration 0.4g/mL.
Rhizoma Dioscoreae Bulbiferae 20% ethanol extract: Rhizoma Dioscoreae Bulbiferae 36g, adds 10 times amount 20% ethanol, reflux, extract, 3 times, each 3h, filters merging filtrate, reclaims ethanol and makes into concentration 0.4g/mL.
Herba Scutellariae Barbatae 20% ethanol extract: Herba Scutellariae Barbatae 36g, adds 10 times amount 20% ethanol, reflux, extract, 3 times, each 3h, filters merging filtrate, reclaims ethanol and makes into concentration 0.4g/mL.
Rhizoma Dioscoreae Bulbiferae acetic acid ethyl acetate extract: Rhizoma Dioscoreae Bulbiferae 36g, Herba Scutellariae Barbatae 36g, adds 10 times amount ethyl acetate, reflux, extract, 3 times, each 3h, filters merging filtrate, reclaims ethyl acetate and makes into concentration 0.4g/mL.
Herba Scutellariae Barbatae acetic acid ethyl acetate extract: Herba Scutellariae Barbatae 36g, adds 10 times amount ethyl acetate, reflux, extract, 3 times, each 3h, filters merging filtrate, reclaims ethyl acetate and makes into concentration 0.4g/mL.
Rhizoma Dioscoreae Bulbiferae acetone extract: Rhizoma Dioscoreae Bulbiferae 36g, adds 10 times amount acetone, reflux, extract, 3 times, each 3h, filters merging filtrate, reclaims acetone and makes into concentration 0.4g/mL.
Herba Scutellariae Barbatae acetone extract: Herba Scutellariae Barbatae 36g, adds 10 times amount acetone, reflux, extract, 3 times, each 3h, filters merging filtrate, reclaims acetone and makes into concentration 0.4g/mL.
Rhizoma Dioscoreae Bulbiferae petroleum ether extract: Rhizoma Dioscoreae Bulbiferae 36g, adds 10 times amount petroleum ether, reflux, extract, 3 times, each 3h, filters merging filtrate, reclaims petroleum ether and makes into concentration 0.4g/mL.
Herba Scutellariae Barbatae petroleum ether extract: Herba Scutellariae Barbatae 36g, adds 10 times amount petroleum ether, reflux, extract, 3 times, each 3h, filters merging filtrate, reclaims petroleum ether and makes into concentration 0.4g/mL.
2.2 experimental technique
2.2.1 the content of diosbulbin B and scutellarin in each extract of high-performance liquid chromatogram determination
Chromatographic column is ultimateTMXB-C18column (4.6 × 250mm, 5 μm); Mobile phase condition is acetonitrile-water=25:75 (v/v), and flow velocity is 1.0mL/min, isocratic elution; Sampling volume is 10 μ L; Column temperature is 25 DEG C; Determined wavelength is 210nm.First get diosbulbin B (DB) appropriate, accurately weighed, add the solution that methanol is made into 1mg/ml, mixing, obtains reference substance solution.DB (1000 μ g/mL) methanol dilution extremely a series of concentration: 7.81,15.63,31.25,62.5,125,250 μ g/mL, mixing, filters with 0.45 μm of organic facies filter membrane, get subsequent filtrate sample introduction, run according to above-mentioned liquid-phase condition, integration tries to achieve peak area value, and result is carried out regression analysis with concentration (μ g/mL) to peak area value and obtained standard curve and correlation coefficient.Secondly, get each extract of Rhizoma Dioscoreae Bulbiferae appropriate, accurately weighed, put in 25ml volumetric flask, add 70% EtOH Sonicate and make it to dissolve, be settled to scale after cooling, filter with 0.45 μm of organic facies filter membrane.Get obtained need testing solution to run according to above-mentioned liquid-phase condition, the peak area value of gained substitutes into above-mentioned standard curve, calculates the content of diosbulbin B in each extract.
Chromatographic column is SepaxHP-C18column (4.6 × 250mm, 5 μm); Mobile phase condition is methanol-water-acetic acid=35:61:4 (v/v/v), and flow velocity is 1.0mL/min, isocratic elution; Sampling volume is 5 μ L; Column temperature is 25 DEG C; Determined wavelength is 335nm.First get scutellarin (SC) appropriate, accurately weighed, add the solution that methanol is made into 1mg/ml, mixing, obtains reference substance solution.SC (1000 μ g/mL) methanol dilution extremely a series of concentration: 15.63,31.25,62.5,125,250,500,1000 μ g/mL, mixing, filters with 0.45 μm of organic facies filter membrane, gets subsequent filtrate sample introduction, run according to above-mentioned liquid-phase condition, integration tries to achieve peak area value, and result carries out regression analysis with concentration (μ g/mL) to peak area value, obtains standard curve and correlation coefficient.Secondly, get each extract of Herba Scutellariae Barbatae appropriate, accurately weighed, be placed in 25mL volumetric flask, add 80% ethanol to about 25mL, supersound extraction 60min, add 80% ethanol after returning to room temperature and be settled to 25mL, after mixing, filter with 0.45 μm of organic facies filter membrane.Get obtained need testing solution to run according to above-mentioned liquid-phase condition, the peak area value of gained substitutes into above-mentioned standard curve, calculates the content of scutellarin in each extract.
2.2.2 tumor-bearing mice anti-tumor experiment (1.2.3 with embodiment 1) in body
2.3 statistical procedures (with 1.3 of embodiment 1)
2.4 experimental result
2.4.1 the anti-tumor activity after other extract administering drug combinations of Rhizoma Dioscoreae Bulbiferae Herba Scutellariae Barbatae
As shown in table 3, compare with model group, each administration group tumor weight average significantly reduces, and all has antitumor action after showing each extract administering drug combinations of Rhizoma Dioscoreae Bulbiferae Herba Scutellariae Barbatae, wherein the strongest with the effect of 70% ethanol extraction and ethyl acetate extract; And 70% ethanol and ethyl acetate extract antitumor action are all obviously better than positive drug 5-Fu's.
On the impact (n=10) of euphorbia egg decoctum solid tumor after table 3 Rhizoma Dioscoreae Bulbiferae extract administering drug combinations different from Herba Scutellariae Barbatae
Compare with model group, * P<0.05, * * P<0.01
2.4.2 the content of diosbulbin B and scutellarin in the different extract of Rhizoma Dioscoreae Bulbiferae Herba Scutellariae Barbatae
As shown in table 4, in 70% ethanol of Rhizoma Dioscoreae Bulbiferae and Herba Scutellariae Barbatae and ethyl acetate extract, the content of diosbulbin B and scutellarin is the highest.
The content of diosbulbin B and scutellarin in the various extract of table 4
Compare with model group, * P<0.05, * * P<0.01
2.4.3 Herba Scutellariae Barbatae ethanol extract increases the anti-tumor activity of Rhizoma Dioscoreae Bulbiferae ethanol extract
As table 5 is visible, compare with model group, the mice of inoculation euphorbia egg decoctum is after giving 70% Rhizoma Dioscoreae Bulbiferae ethanol extract continuously, and tumor representation work reduces (P<0.05), shows that Rhizoma Dioscoreae Bulbiferae 70% ethanol extract is effective to S180 solid tumor; Be used alone with Rhizoma Dioscoreae Bulbiferae ethanol extract and compare, after compatibility Herba Scutellariae Barbatae 70% ethanol extract, tumor weight average significantly reduces (P<0.05), and tumor control rate significantly improves.After giving high dose 20% Rhizoma Dioscoreae Bulbiferae ethanol extract continuously, tumor heavily has certain reduction (P<0.05), be used alone with Rhizoma Dioscoreae Bulbiferae ethanol extract and compare, after compatibility Herba Scutellariae Barbatae 20% ethanol extract, tumor weight average significantly reduces (P<0.05).These all show that Herba Scutellariae Barbatae ethanol extract obviously can work in coordination with the anti-tumor activity increasing Rhizoma Dioscoreae Bulbiferae ethanol extract.
Table 5 Herba Scutellariae Barbatae ethanol extract works in coordination with the anti-tumor activity (n=10) increasing Rhizoma Dioscoreae Bulbiferae ethanol extract
Compare with model group, * P<0.05, * * P<0.01, * * * P<0.001; Compare with the independent medication of Rhizoma Dioscoreae Bulbiferae,
##p<0.01
Embodiment 3 (Chinese medicine composition and active ingredient in Anticancer Activities) thereof
Find that Chinese medicine composition of the present invention has antitumor action by Integral animal experiment, inventor has investigated the active anticancer of this invention Chinese medicine composition and active component thereof further by experiment in vitro.Previous experiments has been found that diosbulbin B is anti-tumor active ingredient (Wang, J.M., Ji main in Rhizoma Dioscoreae Bulbiferae, L.L., Branford-White, C.J., Wang, Z.Y., Shen, K.K., Liu, H., Wang, Z.T., 2012.AntitumoractivityofDioscoreabulbiferaL.rhizomeinviv o.Fitoterapia83,388-394.).Scutellarin is the main active component of Herba Scutellariae Barbatae, and it is also the index components of Herba Scutellariae Barbatae on Chinese Pharmacopoeia simultaneously.Subsequent diosbulbin B and the scutellarin of observing share inhibit activities to tumor growth.
3.1 experiment material
3.1.1 animal subject (1.1.1 with embodiment 1)
3.1.2 test medicine
Diosbulbin B and scutellarin are all purchased from Shanghai Tongtian Biotechnology Co., Ltd..
3.2 experimental techniques (1.2.3 with embodiment 1)
3.3 statistical procedures (with 1.3 of embodiment 1)
3.4 experimental result
As table 6 is visible, DB and SC all has the activity of certain Tumor suppression growth, and wherein DB activity is better than SC.Be used alone when anti-tumor activity after DB and SC use in conjunction is obviously better than.Prompting SC can strengthen the anti-tumor activity of DB.
Table 6DB, SC and drug combination thereof are on the impact (n=10) of S180 sarcoma solid tumor
Compare with model group, * * * P<0.001; Compare with DB,
#p<0.05,
##p<0.01
Embodiment 4 (Chinese medicine composition and the external Anticancer Activities of active component thereof)
Next to Rhizoma Dioscoreae Bulbiferae, Herba Scutellariae Barbatae Chinese medicine composition and active component diosbulbin B, the anti tumor activity in vitro of scutellarin compositions is verified further.
4.1 experiment material
4.1.1 cell strain source
Human pancreas cancer BXPC-3, breast carcinoma MCF-7, pulmonary carcinoma A549, gastric cancer SGC-7901, hepatocarcinoma Hep3B, bladder cancer T24, human ovarian cancer HO8910, colon cancer LS174 and CCL229, carcinoma of prostate PC-3, hunman esophageal carcinoma Eca-109 cells, skin carcinoma A431, thyroid carcinoma SW-579, leukemia K 562, s strain are all purchased from Shanghai Inst. of Cytobiology, Chinese Academy of Sciences.
4.1.2 main agents
DEME, RPMI1640 culture medium (GIBCO company), hyclone (GIBCO company), dimethyl sulfoxide (DMSO) (GIBCO company), MTT (Sigma company), 5-Fu (Shanghai nation one-tenth), other reagent is domestic analytical pure etc.
4.1.3 key instrument equipment
Laminar flow super-clean bench; CO2 gas incubator, purchased from Thermo company; Inverted phase contrast microscope, purchased from Olympus company; Microplate reader, purchased from Biotek company.
4.2 experimental technique
Cell proliferation experiment (MTT): use 0.25% trypsin digestion cell, 1000rpm is centrifugal, with containing 5% active carbon inactivated serum, without the high sugared culture fluid 37 DEG C of phenol red DMEM, 95%CO
2cultivate 48h (to get rid of the interference of endogenous hormones), namely cell enters exponential phase, uses 0.25% trypsin digestion cell, with 2 × 10
3individual/hole inoculating cell is on 96 well culture plates, and every pore volume 180 μ L, 24h cell attachment, often row establish 5 multiple holes, establish solvent control, 5-Fu positive control, Experimental agents group respectively, continue to observe and cultivate 48h after dosing.
Colour generation and colorimetric: after 48h is cultivated in dosing, take out culture plate, every hole adds MTT solution (5mg/mL) 20 μ L, 37 DEG C, 5%CO
2continue to cultivate 4h to stop cultivating.Culture supernatant in hole is abandoned in careful suction, every hole adds 150 μ LDMSO vibration 10min, and crystal is fully dissolved, and selects 490nm at each hole absorption value (OD of the upper mensuration of automatic microplate reader (BIO-TEK), be directly proportional to living cells quantity), calculate inhibitory rate of cell growth.Inhibitory rate of cell growth=(control wells OD value-experimental port OD value)/control wells OD value × 100%.
4.3 experimental result
From table 7-11, Chinese medicine composition 80% ethanol extract of the present invention, diterpenoid-lactone active component diosbulbin B (DB) in Rhizoma Dioscoreae Bulbiferae, all human pancreas cancer BXPC-3 can be significantly suppressed with Herba Scutellariae Barbatae active component scutellarin (SC), breast carcinoma MCF-7, pulmonary carcinoma A549, gastric cancer SGC-7901, hepatocarcinoma Hep3B, bladder cancer T24, ovarian cancer HO8910, colon cancer LS174 and CCL229, carcinoma of prostate PC-3, hunman esophageal carcinoma Eca-109 cells, skin carcinoma A431, thyroid carcinoma SW-579, the growth of leukemia K 562 or s strain, point out this Chinese medicine composition, diosbulbin B and scutellarin can be used for treating human pancreas cancer, breast carcinoma, pulmonary carcinoma, gastric cancer, hepatocarcinoma, bladder cancer, ovarian cancer, colon cancer, carcinoma of prostate, the esophageal carcinoma, skin carcinoma, thyroid carcinoma, the kinds cancer such as leukemia or cervical cancer.Simultaneously, the present invention has also investigated the anticancer effect of active ingredient compositions, found that such as scutellarin in the flavones ingredient contained by Herba Scutellariae Barbatae, with significantly can strengthen the antitumaous effect of diosbulbin B to above multiple cancerous cell during diosbulbin B compatibility, and the active anticancer after compatibility is obviously better than and is used alone, the flavones ingredient scutellarin of prompting contained by Herba Scutellariae Barbatae can with contained Diterpenes constituents diosbulbin B in Rhizoma Dioscoreae Bulbiferae can compatibility efficient application in treating above Cancerous disease.
Table 7 compositions and active component thereof are on the impact of people cancer BXPC-3, MCF-7, A549 inhibitory rate of cell growth (%)
Compare with solvent control group, * P<0.01; Compare with DB,
#p<0.05
Table 8 compositions and active component thereof are on the impact of people cancer SGC-7901, Hep3B, T24 inhibitory rate of cell growth (%)
Compare with solvent control group, * P<0.01; Compare with DB,
#p<0.05
Table 9 compositions and active component thereof are on the impact of people cancer HO8910, LS174, PC-3 inhibitory rate of cell growth (%)
Compare with solvent control group, * P<0.01; Compare with DB,
#p<0.05
Table 10 compositions and active component thereof are on the impact of people cancer CCL229, Eca-109, A431 inhibitory rate of cell growth (%)
Compare with solvent control group, * P<0.01; Compare with DB,
#p<0.05
Table 11 compositions and active component thereof are on the impact of people cancer SW-579, K562, Hela inhibitory rate of cell growth (%)
Compare with solvent control group, * P<0.01; Compare with DB,
#p<0.05
Embodiment 5 (preparation of fluid extract)
Get Rhizoma Dioscoreae Bulbiferae, each 250g of Herba Scutellariae Barbatae, beat powder, use 70% ethanol as solvent, dipping 48h, slowly percolation, collect the liquid 425mL that just filters, another device is preserved; Continue percolation, to percolate close to colourless, collect continuous liquid of filtering, be concentrated into paste at 55 DEG C, add the liquid 425mL that just filters and mix, with 70% ethanol dilution to 500mL, standing 3d, filtration, to obtain final product.
Embodiment 6 (preparation of tablet)
Get Rhizoma Dioscoreae Bulbiferae, each 250g of Herba Scutellariae Barbatae, beat powder, cross 100 mesh sieves.Spray into appropriate 80% ethanol, add 17.5% microcrystalline Cellulose, add 7.5% microcrystalline Cellulose and 20% starch wherein, make granule, dry, additional 10% microcrystalline Cellulose and 1% magnesium stearate, mixing tabletting.
Embodiment 7 (preparation of drop pill)
Get Rhizoma Dioscoreae Bulbiferae, each 250g of Herba Scutellariae Barbatae, beat powder, with 20% alcohol reflux 3 times, each 2h, reclaim ethanol and obtain extractum, taking PGE4000 as substrate, is 1:1.5 with the ratio of medicine, and dimethicone is condensing agent, dripping.
Embodiment 8 (preparation of capsule)
Get Rhizoma Dioscoreae Bulbiferae, each 250g of Herba Scutellariae Barbatae, beat powder, extract 3 times by ethyl acetate backflow, each 2h, reclaim ethyl acetate and obtain extractum, dry, pulverize, cross 100 mesh sieves, add 5% magnesium stearate, mixing loads hard capsule.
Many aspects involved in the present invention have been done and have as above been set forth.It is to be understood, however, that put before not departing from spirit of the present invention, those skilled in the art can carry out equivalent change and modification to it, and described change and modification fall into the coverage of the application's claims equally.
Claims (13)
1. an antineoplastic Chinese medicine composition, it is characterized in that, described Chinese medicine composition is made up of the crude drug of following weight percents: Rhizoma Dioscoreae Bulbiferae 10% ~ 70%, Herba Scutellariae Barbatae 30% ~ 90%, and described Chinese medicine composition is the full powder suspension of Rhizoma Dioscoreae Bulbiferae and Herba Scutellariae Barbatae, or its 20% ~ 80% ethanol extraction, ethyl acetate extract, acetone extract or ligroin extraction.
2. Chinese medicine composition as claimed in claim 1, is characterized in that, the Rhizoma Dioscoreae Bulbiferae composition in described Chinese medicine composition is containing diosbulbin B 1% ~ 17%, and the Herba Scutellariae Barbatae composition in described Chinese medicine composition is containing scutellarin 1% ~ 15%.
3. the application of the Chinese medicine composition described in claim 1 or 2 in the medicine or health product of preparation control cancer of pancreas, breast carcinoma, pulmonary carcinoma, gastric cancer, hepatocarcinoma, bladder cancer, ovarian cancer, colon cancer, carcinoma of prostate, the esophageal carcinoma, skin carcinoma, thyroid carcinoma, leukemia or cervical cancer.
4. the purposes of a Herba Scutellariae Barbatae extract, it is characterized in that, described Herba Scutellariae Barbatae extract can be used for the anti-tumor activity improving Rhizoma Dioscoreae Bulbiferae extract, and described Herba Scutellariae Barbatae extract and Rhizoma Dioscoreae Bulbiferae extract are respectively Herba Scutellariae Barbatae 20% ~ 80% ethanol extraction and Rhizoma Dioscoreae Bulbiferae 20% ~ 80% ethanol extraction.
5. purposes as claimed in claim 4, it is characterized in that, the weight ratio of described Herba Scutellariae Barbatae extract and Rhizoma Dioscoreae Bulbiferae extract is 1:2 ~ 2:1.
6. purposes as claimed in claim 4, is characterized in that, described Herba Scutellariae Barbatae extract is containing scutellarin 1% ~ 6%, and described Rhizoma Dioscoreae Bulbiferae extract is containing diosbulbin B 1% ~ 7%.
7. an antineoplastic Chinese medicine composition, is characterized in that, described Chinese medicine composition is grouped into by the one-tenth of following weight ratio: Rhizoma Dioscoreae Bulbiferae 20% ~ 80% ethanol extraction: Herba Scutellariae Barbatae 20% ~ 80% ethanol extraction=1:2 ~ 2:1.
8. Chinese medicine composition as claimed in claim 7, is characterized in that, described Rhizoma Dioscoreae Bulbiferae extract is containing diosbulbin B 1% ~ 7%, and described Herba Scutellariae Barbatae extract is containing scutellarin 1% ~ 6%.
9. the application of the Chinese medicine composition described in claim 7 or 8 in the medicine or health product of preparation control cancer of pancreas, breast carcinoma, pulmonary carcinoma, gastric cancer, hepatocarcinoma, bladder cancer, ovarian cancer, colon cancer, carcinoma of prostate, the esophageal carcinoma, skin carcinoma, thyroid carcinoma, leukemia or cervical cancer.
10. a purposes for scutellarin, is characterized in that, described scutellarin can be used for the anti-tumor activity improving diosbulbin B.
11. purposes as claimed in claim 10, is characterized in that, the weight ratio of described scutellarin and diosbulbin B is 1:1 ~ 2:1.
12. 1 kinds of antitumor medicine compositions, is characterized in that, described Chinese medicine composition is grouped into by the one-tenth of following weight ratio: diosbulbin B: scutellarin=1:1 ~ 1:2.
The application of 13. pharmaceutical compositions according to claim 12 in the medicine or health product of preparation control cancer of pancreas, breast carcinoma, pulmonary carcinoma, gastric cancer, hepatocarcinoma, bladder cancer, ovarian cancer, colon cancer, carcinoma of prostate, the esophageal carcinoma, skin carcinoma, thyroid carcinoma, leukemia or cervical cancer.
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| CN105362691A (en) * | 2015-12-02 | 2016-03-02 | 钊桂英 | Traditional Chinese medicine composition mainly for treating leukemia and preparation method |
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| CN108836963B (en) * | 2018-05-18 | 2020-06-05 | 山东省中医药研究院 | Application of diosbulbin B in preparation of drug for treating nodular goiter |
| CN113491771A (en) * | 2020-03-18 | 2021-10-12 | 苏州大学 | Application of small molecular compound in preparation of GLUT5 inhibitor for uptake and transportation of fructose |
| CN114306349A (en) * | 2022-01-07 | 2022-04-12 | 北京蕴汇医药科技有限公司 | Application of chenodeoxycholic acid or derivative thereof in preparation of drugs for synergistically preventing or treating diseases related to EGFR (epidermal growth factor receptor) |
| CN115444900A (en) * | 2022-04-16 | 2022-12-09 | 复旦大学 | Traditional Chinese medicine composition for treating pancreatic cancer and preparation method and application thereof |
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