CN1486712A - Extractive of Chinese medicinal material rabdosia as antitumor medicine and its prepn process - Google Patents
Extractive of Chinese medicinal material rabdosia as antitumor medicine and its prepn process Download PDFInfo
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Abstract
The present invention relates to Chinese medicine preparation, and is especially composition of effective rabdosia parts and its preparation process. Rabdosia as material is processed through crushing, alcohol extraction, concentration and extraction to obtain coarse rabdosia total terpene extractive; coarse rabdosia total terpene extractive is then re-dissolved, eluted and freeze dried or spray dried to obtain rabdosia total terpene powder; and the rabdosia total terpene powder is finally prepared into injection or oral liquid. The medicine of the present invention may be used in treating liver cancer, oesophagus cancer, stomach cancer, intestinal cancer, breast cancer, lung cancer, melanoma and other malignant tumor, and has obvious curative effect and no toxic side effect.
Description
Technical field
The present invention relates to a kind of the antineoplastic Chinese medicine that has, especially Rabdosia rubescens effective part extract and this extract is made the production method of antitumor Chinese medicine preparation.
Background technology
Rabdosia rubescens, have another name called Rabdosia rubescens, formal name used at school Tabdosia rubescens (hemsl.), be Labiatae Rabdosia perennial herb or semishrub plant, there is wide model distribute on China Henan, Hebei, Shanxi, Sichuan and other places, includes in " first one of Chinese pharmacopoeia version in 1977 and " Ministry of Public Health Chinese crude drug standard " version in 1992.
The Rabdosia rubescens sweet-bitter flavor, cold nature has heat-clearing and toxic substances removing, anti-inflammatory analgetic, the effect such as invigorate blood circulation that is good for the stomach, the document record can be used for the treatment of diseases such as acute and chronic tonsillitis, pharyngolaryngitis, stomatitis.Medicines such as now existing commercially available Dong ling cao tablet, Rabdosia rubescens syrup are mainly treated oral inflammation.In addition, Rabdosia rubescens also has has low dose of the inhibition to staphylococcus aureus, Diplococcus pneumoniae, group B streptococcus etc., and the effect that heavy dose is killed and wounded waits other effect.
The discovered in recent years Rabdosia rubescens also has anti-tumor activity.The ethanol extract of Rabdosia rubescens has obvious inhibitory action to the Hela cell; Ehrlich carcinoma is injected under 10g/Kg concentration, and tumor control rate can reach 60%.Clinical experiment shows, Rabdosia rubescens is to the esophageal carcinoma, carcinoma of gastric cardia, and the treatment of hepatocarcinoma etc. also has certain curative effect, and showing as appetite increases, pain relief, the tumor body dwindles.
By retrieval, find that scientific and technical literature discloses the pharmacology of some research Rabdosia rubescens extraction processes and Rabdosia rubescens and Rabdosia rubescens is applied to the article of clinical treatment, now takes passages as follows:
1, Rabdosia rubescens leaf Study on extraction " time precious traditional Chinese medical science traditional Chinese medicines " .2001,12 (4) .[digests] purpose: inquire into the optimum condition that the Rabdosia rubescens leaf is extracted with 95% ethanol.Method: orthogonal test, in measure extracting with the HPLC method in the Rabdosia rubescens content of first element be that index is carried out, extraction time 4h (extracting twice) adds 95% ethanol than 10: 1,70 ℃ of extraction temperature.Conclusion: it is better that this method is extracted Rabdosia rubescens leaf effect.
2, the new technology of medium pressure column chromatography method extraction separation rubescensine A " Heilungkiang medicine " .2001,14 (1) .[digests] adopt medium pressure column chromatography method extraction separation rubescensine A, it is fast to have an elution speed, the separation efficiency height, the solvent nontoxic pollution-free, advantage such as safe, inexpensive, as to be easy to get.Reaching 99.28% with tlc scanning determination content behind recrystallization, is a kind of simple and easy to do method.
3, rubescensine A mutation Journal of Sex Research " canceration. distortion. sudden change " [digest] purpose: the antimutagenic effect of inquiring into rubescensine A.Method: adopt Salmonella reversion test and PCEMNR (PCE) micronucleus test.The result: rubescensine A is not adding under the S9 condition, TA98 and TA100 back mutation had the obvious suppression effect, its the highest suppression ratio reaches 89.1 and 80.2% respectively, to by the inductive mouse bone marrow cells PCE microkernel incidence of cyclophosphamide (CP) significant antagonism (P＜0.01) being arranged also.Conclusion: Rabdosia rubescens, the first element has antimutagenic activity under this experiment condition.
4, the anticancer molecular pathology of rubescensin and clinical practice strategy " Guangdong pharmacy " .2000,10 (4) .[digests] rubescensin is the novel compound that China at first finds from traditional herbal medicine Rabdosia rubescens (Raddosia rubescens), be a PTS with specially good effect low toxicity, especially effective to the esophageal carcinoma and carcinoma of gastric cardia.It is the newest fruits of research in recent years about rubescensin that this paper has summarized, inquired into the molecular mechanism of this medicine antitumaous effect from molecular pharmacology and oncobiology angle, the relation of active anticancer and compound structure, and the synergistic function of rubescensin and chemotherapeutics such as bleomycin.
5, rubescensine A is to scavenging action " Henan medical research " .1999 of reactive oxygen free radical, 8 (2) .[digests] scavenging action of research rubescensine A reactive oxygen free radical that different model systems are produced.Revolve the resonance free radical and revolve trapping technique and chemical light method at method free radical detection place.The result: two kinds of methods detect all to be found obviously to remove superoxide radical that xanthine/xanthine oxidase enzyme system produces and the hydroxy radical in the Fenton reaction by Orid (OH), Orid170mol/l is to O
-2Be respectively 49.4% and 75.2% with the clearance rate of-OH.Orid is to the obvious scavenging action of the same demonstration of the bright generation reactive oxygen free radical of polymorphonuclear leukocyte (PMN) respiratory burst.
6, the electronic structure of rubescensine A and acetyl derivative thereof and active anticancer " Zhengzhou University's journal ": from section's version 1998,30 (1) [digest] this paper has carried out quantum chemistry calculation research to rubescensine A and acetyl derivative thereof with the MNDO method on the VAX8350 computer, track and energy level thereof have been provided, charge density, and the relation of its electronic structure and active anticancer discussed.
7, the progress of Rabdosia rubescens " Chinese Pharmaceutical Journal " .1998,33 (10) .[digests] purpose: Rabdosia rubescens is summarized in the progress of aspects such as chemistry, pharmacology over nearly 30 years.Method: consult medical Chinese material and U.S. CA data study achievement in the past in 1997, summarized main chemical compositions, extraction process, determination, pharmacology and the clinical practice of Rabdosia rubescens.The result: main component is a kaurene class diterpene, and the Rabdosia rubescens in the different places of production contains two different terpene components, and rubescensine A has many-sided physiologically active.Conclusion: the antitumaous effect that Rabdosia rubescens is certain, and toxicity, side effect is little, is expected to be developed to new anticarcinogen, brings benefit to the mankind.
8, the electronic structure of rubescensine B and active anticancer " Henan Medical Univ.'s journal " .1998,33 (3) .[digests] adopt the MNDO method, on the VAX8350 computer, rubescensine B is carried out quantum chemistry calculation.Molecular orbit and energy level thereof have been provided, charge density, bond order.The result shows: the part of atoms in the active anticancer district alpha-methylene Ketocyclopentane structure has been formed the π LUMO track of easily accepting electronics, easily in this zone nucleophilic addition takes place.
9, different Study on extraction process " Chinese herbal medicine " .1997 of Rabdosia rubescens, 28 (7) [digests] are studied the different extraction processes of Rabdosia rubescens, and are that index has been carried out assay to it respectively with the rubescensine A.The result shows, the rubescensine A content that broken extraction method is extracted than other extraction method is for the highest.
10, the Rabdosia rubescens treatment esophageal carcinoma, carcinoma of gastric cardia 54 routine clinical observation on the therapeutic effect " Henan oncology's magazine " .1993,6 (3) .[digests] treat esophageal neoplasm, cardia tumor with Rabdosia rubescens, and observe its curative effect.
11, Crystal Structure of Oridonin " structural chemistry " .1992,11 (6) .[digests] Crystal Structure of Oridonin is analyzed, and introduce its application as anticarcinogen.
12, the source new drugs of Rabdosia rubescens is inquired into " Henan science " .1989,7 (3) .[digests] extract the tetracyclic diterpene that obtains from Rabdosia rubescens, can be used as antibacterial, antineoplastic medicine source.
13, the molecular configuration of cancer therapy drug rubescensine A research " chemical journal " .1992,50 (5) .[digests] analyzed the molecular configuration of rubescensine A, its anti-cancer properties is studied.
14, Oridonin derivative and preparation method thereof, Chinese patent, application number, 99101179, the applicant, the present invention of Zhengzhou University's [digest] is under the prerequisite of not destroying the rubescensine A active center, by changing the tension force and the dissolubility of the outer methylene Ketocyclopentane of α-ring, provide in general formula 1: R ↑ [1] is alkyl or the acyl group of H or C1～C12, or is glucosyl group or galactosyl or xylosyl, R ↑ [2] are H or glucosyl group or galactosyl or xylosyl, or mannose group; In general formula 2: R ↑ [1] is alkyl or the acyl group of H or C1～C12; R ↑ [2], R ↑ [3] are respectively the alkyl of C2～C18, or are isopropylidene, benzene methene base.Shown Oridonin derivative: this Oridonin derivative has higher antitumor cell activity.Simultaneously, the present invention also provides the preparation method of above-claimed cpd.
Recognize from above-mentioned result for retrieval, the research that medical institutions and Chinese medicine scientific research department carry out Rabdosia rubescens is the chemical constitution and the antitumaous effect of rubescensine A, rubescensine B, Oridonin derivative, kaurene class diterpene, and the structure that provides Rabdosia rubescens part terpenoid in the document is rubescensine A and rubescensine B.
The rubescensine A rubescensine B
But all do not disclose other effective ingredient of Rabdosia rubescens in the document, do not provide the application of other effective ingredient of Rabdosia rubescens on the treatment malignant tumor yet.
Technology contents
The inventor after research and testing, Rabdosia rubescens is extracted with various method, analyze the composition of extract, the terpenoid in the Rabdosia rubescens extract particularly, except obtaining rubescensine A, rubescensine B, Oridonin derivative, also obtain some and be different from the chemical substance of open source literature, these chemical substances have been used for the treatment of malignant tumor, received better effect.
Technical scheme of the present invention is as follows:
Have antineoplastic Rabdosia rubescens effective part extract and comprise the total terpene of Rabdosia rubescens, it comprises rubescensine A (rubescensinA) and derivant, rubescensine B (rubescensin B), rubescensine C (rubescensin C), rubescensine D (rubescensin D), rubescensine E (rubescensin E) and rubescensine H diterpene-kind compound and triterpenoid compound such as oleanolic acid and ursolic acid such as (rubescensin F).The total terpene content of Rabdosia rubescens preferably is not less than 55% more than 50% in this extract.Concrete content can be determined by chemical method or Instrumental Analysis.
The inventor finds also that by structural analysis rubescensin all is a kaurene skeleton tetracyclic diterpene compounds, is its physiologically active center with ring outer methylene conjugated Ketocyclopentane structure in its structure, and driffractive ring or methylene be saturated then can to make its effect inefficacy.Some other group plays a part enhanced activity, and owing to the hydrogen bond with the formation of 6-position hydroxyl and 15-position carbonyl has higher activity, these characteristic open source literatures do not appear in the newspapers as rubescensine A.
The invention provides the preparation method of the total terpene of above-mentioned Rabdosia rubescens, these preparation methoies more can obtain comprehensive chemical compound effectively compared with existing extracting method.
The present invention also provides the total terpene of Rabdosia rubescens to make the preparation of pharmaceutically commonly using, and the application of the active compound of conduct treatment malignant tumor, and these malignant tumor comprise hepatocarcinoma, the esophageal carcinoma, gastric cancer, intestinal cancer, breast carcinoma, pulmonary carcinoma and melanoma etc.
The above-described preparation of pharmaceutically commonly using comprises injection and oral agents, and wherein oral formulations comprises capsule, soft capsule, tablet, granule, slow releasing agent, oral liquid etc.; Injection comprises various aqueous injection and injectable powder.
Above-described method with antineoplastic Rabdosia rubescens effective part extract (the total terpene of Rabdosia rubescens) extraction may further comprise the steps:
1, pulverize the back Rabdosia rubescens 3-4 time with alcoholic solution or double solvents reflux, extract,, solvent load is 5-8 a times of raw material, obtains the Rabdosia rubescens crude extract;
2, merge crude extract, concentrating under reduced pressure reclaims solvent, is concentrated into relative density 1.05-1.10;
3, with the extract petroleum ether extraction after above-mentioned the concentrating, slough fat and pigment, residue extracted with diethyl ether 3-4 time;
4, merge ethereal extract, reclaim ether, get the total terpene crude extract of Rabdosia rubescens;
5, the total terpene crude extract of Rabdosia rubescens being dissolved with low-carbon alcohols, cross macroporous adsorptive resins, is that mobile phase is carried out eluting with gradient 10-80% alcoholic solution.Go out product with TLC method monitoring stream, collect the outflow product that contains the total terpene of Rabdosia rubescens;
6, merge eluent, concentrating under reduced pressure is removed solvent, and freezing or spray drying is to powder, the total terpene of Rabdosia rubescens.As adding the crystallization of hot acetone solution weight again this moment, can obtain purer monomeric compound.
In the said extracted step 1, the extraction of the total terpene of Rabdosia rubescens can be taked merceration extraction method, percolation extraction method, ultrasonication extraction method, reflux extraction etc.Used extraction solvent has Different concentrations of alcohol, ether, acetone or their mixture etc.The preferred reflux extraction of the present invention, adopting solvent is the 70-95% alcoholic solution.
In the said extracted step 3, the decolouring of Rabdosia rubescens crude extract can be adopted methods such as organic solvent extraction and activated carbon adsorption, and preferred petroleum ether extraction method when small lot of the present invention is extracted during production in enormous quantities, is taked the activated carbon adsorption decoloring method.
In the said extracted step 5, column chromatographic isolation and purification, can take with neutral and polar macroporous adsorption resin, silica gel, aluminium oxide, ion exchange resin is that inserts adsorbs, the preferred macroporous adsorbent resin of the present invention.Used eluent can be ethanol, methanol, chloroform, acetone, petroleum ether or their mixed liquor and aqueous solution.Preferred alcohol aqueous solution of the present invention.When concentration of alcohol is increased to the product that can obtain after 50% based on rubescensine A, and pigment and other impurity are adsorbed removal to a great extent.
In the said extracted step 6, the preferred temperature of concentrating under reduced pressure is 50-60 ℃, and vacuum is 400-600Mpa.For drying condition, can adopt vacuum drying, atmospheric steam drying, lyophilization and spray drying.Preferably freeze drying of the present invention and spray drying.
According to technical scheme of the present invention, its form of Chinese drug can be injection and oral agents, and wherein oral formulations comprises capsule, soft capsule, tablet, granule, slow releasing agent, oral liquid etc.; Injection comprises various aqueous injection and injectable powder.In preparation during oral formulations, the adjuvant of selecting for use can be starch, carboxymethyl cellulose, cyclodextrin, sucrose, magnesium stearate, Polyethylene Glycol, etc. conventional filler, can adopt the conventional preparation technology of pharmaceutics; When the preparation ejection preparation, can adopt conventional filleies such as adding liposome, Polyethylene Glycol, medicinal alcohol, deoxycholic acid, cholesterol, can adopt the conventional preparation technology of pharmaceutics, so this ominous stating.
The present invention is injection preferably, particularly liposome aqueous injection and lyophilized injectable powder.
For guaranteeing the quality of medicine, need in process of production the main effective ingredient of Rabdosia rubescens is monitored.The present invention proposes the feasible method of quality control of a cover.
Thin layer chromatography (hole C) method:, can control to main active in extracting solution and the column chromatography eluent by TLC.The TLC condition: developing solvent is a chloroform: acetone=8: 1-2, develop the color with 50% sulphuric acid ethanol.
High performance liquid chromatography (HPLC) method:, adopt single active constituent content in the external standard method final products by HPLC.The HPLC condition:
Equipment: Waters 600, Waters 996 pumps, Photodiode Array Detector detector;
Chromatographic column: Dikma, Diamonsil C18,4.6mm*25cm;
Mobile phase: methanol: water=60: 40 (V: V);
Detected temperatures: room temperature; Detect wavelength: 240nm; Flow velocity: 0.8ml/min.
The present invention compared with prior art, its outstanding substantive distinguishing features and obvious improvement is:
1, with new extracting method, can be all total terpene of Rabdosia rubescens be extracted, obtain some and be different from the chemical substance of open source literature.
2, the chemical compound complete function of the total terpene of Ling Ge can be brought into play the effect of its anti-various cancerous cell, and the treatment effectiveness of medicine is improved.
3, anticancer spectrum is wide, high-efficiency low-toxicity; Crude drug source is abundant, low price; Effective ingredient is clear and definite; Controllable product quality.
4, the quality of medicine more can guarantee.
5, production technology is easy, adopts the column chromatographic isolation and purification technology, is easier to suitability for industrialized production.
Description of drawings
Fig. 1 is rubescensine A reference substance HPLC figure.
Fig. 2 is the rubescensine A canonical plotting.
As can be seen from Figure 1, by HPLC, adopt external standard method can measure single active constituent content in the final products, such as winter icepro among the figure The plain reference substance purity of grass first does not have other impurity (Rt=3.2 is solvent peak) greater than 99%.
Fig. 2 measures the total terpene content of Rabdosia rubescens and can adopt uv-spectrophotometric (UV) method as can be known: by the UV method, measure finally and produce Total terpene content in the thing. The employing Oridonin is standard items, the preparation variable concentrations, and take concentration as abscissa, absorbance is ordinate Mapping, the drawing standard curve. Get regression equation: Y=20.156X+0.0119. Accurately weighed test sample dissolves the ultraviolet branch then Light luminosity instrumentation absorbance, the substitution regression equation calculates content.
The specific embodiment
Preparation embodiment one:
Get Rabdosia rubescens (containing stem and leaf) medical material, pulverize, sieve, add 95% ethanol of 6 times of volumes, reflux, extract, 3 hours.Repeat to extract twice, merge extractive liquid.Concentrating under reduced pressure reclaims solvent, is concentrated into relative density 1.05-1.10.With the petroleum ether extraction twice of the extract after above-mentioned the concentrating, slough fat and pigment with 3 times of volumes.Residue extracted with diethyl ether 3-4 time merges ethereal extract, reclaims ether, and residue dissolves with 90% ethanol alcohol, and supernatant is crossed macroporous adsorptive resins, is that mobile phase is carried out eluting with gradient 10-80% alcoholic solution.Collect the outflow product that contains the total terpene of Rabdosia rubescens with the monitoring of TLC method.Merge eluent, concentrating under reduced pressure, the dry total terpene of Rabdosia rubescens that gets.
Preparation embodiment two:
Get Rabdosia rubescens (containing stem and leaf) medical material, pulverize, sieve, add the ether of 5 times of volumes, reflux, extract, 3 hours.Repeat to extract twice, merge extractive liquid.Concentrating under reduced pressure reclaims solvent, is concentrated into relative density 1.05-1.10.With the extract activated carbon adsorption after above-mentioned the concentrating, slough fat and pigment.Residue extracted with diethyl ether 3-4 time merges ethereal extract, reclaims ether, and residue dissolves with 90% ethanol alcohol, and supernatant is crossed macroporous adsorptive resins, is that mobile phase is carried out eluting with gradient 10-80% alcoholic solution.Collect the outflow product that contains the total terpene of Rabdosia rubescens with the monitoring of TLC method.Merge eluent, concentrating under reduced pressure, the dry total terpene of Rabdosia rubescens that gets.
Application Example one: the total terpene of Rabdosia rubescens is to the inhibitory action of human breast cancer cell MCF7
Human breast cancer cell MCF7 places the RPMI RPMI-1640 that contains 10% hyclone, and 37 ℃, the 5%CO2 saturated humidity is cultivated, and goes down to posterity once in the 2nd day.Using the culture medium dilute liquid medicine during experiment, is 50 μ g/ml with concentration finally.The different tumor cells that grow into logarithmic (log) phase are digested to cell suspension with Digestive system, and with every hole 10
3-10
4Individual cell, the amount of 200 μ l is inoculated into 96 well culture plates.After 24 hours, remove the culture medium of cultivating in the plate hole, be changed by fresh bioactive peptide medicinal liquid, every hole 200 μ l.Dosing was cultivated respectively 3 days after handling, and every then hole adds the MTT solution of 20 μ l, and 37 ℃ are continued to cultivate 4 hours.The careful supernatant of removing culture in the hole, every then hole adds the DMSO of 200 μ l, and concussion is 20 minutes on the horizontal shaking table, and the assurance crystallization is fully dissolved.Detect the absorbance value in each hole at the 490nm place, calculate suppression ratio.The result shows, at drug level during greater than 40 μ g/ml, the total terpene of Rabdosia rubescens to the suppression ratio of human breast cancer cell MCF7 all greater than 50%.When drug level was 50 μ g/ml, suppression ratio was 92%.
Application Example two: the total terpene of Rabdosia rubescens is to the inhibitory action to the human lung cancer cell A549
The total terpene of Rabdosia rubescens is dissolved with DMSO, be made into the 20mg/ml mother solution ,-20 ℃ of preservations are standby.Diluting with culture medium during experiment, is 100 μ g/ml with concentration finally.The different tumor cells that grow into logarithmic (log) phase are digested to cell suspension with Digestive system, and with every hole 10
3-10
4Individual cell, the amount of 200 μ l is inoculated into 96 well culture plates.After 24 hours, remove the culture medium of cultivating in the plate hole, be changed by fresh bioactive peptide medicinal liquid, every hole 200 μ l.Dosing was cultivated respectively 3 days after handling, and every then hole adds the MTT solution of 20 μ l, and 37 ℃ are continued to cultivate 4 hours.The careful supernatant of removing culture in the hole, every then hole adds the DMSO of 200 μ l, and concussion is 20 minutes on the horizontal shaking table, and the assurance crystallization is fully dissolved.Detect the absorbance value in each hole at the 490nm place, calculate suppression ratio.The result shows that the suppression ratio to human lung carcinoma cell when drug level is 40 μ g/ml is 74%, and is certain dose-effect relationship.
Application Example three: the total terpene of Rabdosia rubescens is to the inhibitory action of other tumor cells
Employing is carried out the inhibitory action experiment of the total terpene of Rabdosia rubescens to human esophagus cancer cell EC9706, human liver cancer cell HepG2 and mouse melanin tumor cell K1735M2 with embodiment two, three similar approach.The result shows that the suppression ratio to esophageal cancer cell EC9706, hepatoma carcinoma cell HepG2 and mouse melanin tumor cell K1735M2 when drug level is 40 μ g/ml is respectively 61%, 67 and 78%, and is certain dose-effect relationship.
Application Example four: the total terpene of Rabdosia rubescens is to the influence of ehrlich ascites tumor mice
Under aseptic condition, from mouse peritoneal ascites, get tumor, dilution, sterilization inoculation mouse peritoneal behind the mixing, every 0.2ml contains oncocyte 1 * 10 approximately
6Individual.Be divided into matched group (normal saline) behind the 24h at random, the total terpene low dose of cyclophosphamide group and Rabdosia rubescens (10mg/kg) group, middle dosage (20mg/kg) group, heavy dose of (40mg/kg) group.Intravenously administrable, totally 7 days, the mean survival time of observing every group of mice, calculates increase in life span at every day 1 time.Experimental result shows that the total terpene low dose of Rabdosia rubescens (10mg/kg) group, middle dosage (20mg/kg) group, heavy dose of (40mg/kg) group all can obviously prolong ehrlich ascites tumor mice life span, and increase in life span is all greater than 50%.
Application Example five: the total terpene of Rabdosia rubescens is to the inhibitory action of mouse entity tumor
According to embodiment five similar approach, adopt mice right hind subcutaneous vaccination S
180The solid tumor cell suspension is pressed 10mg/kg body weight intravenously administrable with the total terpene of above-mentioned Rabdosia rubescens, and the result shows S
180The suppression ratio of solid tumor is 68.3%.
Application Example seven: preparation tablets
According to prescription, the total terpene of Rabdosia rubescens is pulverized, sieve, take by weighing 40g, add starch, lactose mix homogeneously, add 60% ethanol system soft material, granulate, drying, granulate adds magnesium stearate, and mixing is pressed into 1000, and sugar coating is drying to obtain.Every contains pharmaceutical composition 40mg.
Application Example eight: capsule preparation
Take by weighing 40g and pulverize, the total terpene of Rabdosia rubescens that sieves and handle adds starch and magnesium stearate, mix homogeneously, and filled capsules, every contains compositions 40mg.
Application Example nine: granule preparation
Take by weighing pulverizing, the pharmaceutical composition, cane sugar powder, dextrin and an amount of ethanol that sieve and handle, stirring and evenly mixing is granulated drying, granulate, packing, finished product.The heavy 5g of every bag.
Application Example ten: soft capsule preparation
Take by weighing pulverizing, the pharmaceutical composition, the vegetable oil that sieve and handle fully mix, and the colloid mill mill is even, places full-automatic encapsulating machine filling, drying, promptly.Every contains pharmaceutical composition 40mg.
Application Example 11: lipidosome injection preparation
According to prescription, take by weighing pharmaceutical composition, soybean lecithin, cholesterol, sodium deoxycholate, mix homogeneously adds absolute ether, and thin film evaporation instrument rotary evaporation is until forming homogeneous film.Add buffer solution then, continue aquation 2h, ultrasonic, ultrafiltration.Add the injection water, fill.
Claims (4)
1, a kind of Chinese medicine Rabdosia rubescens effective site antitumor drug extract, it is characterized in that: its effective part extract is the total terpene of Rabdosia rubescens, it comprises rubescensine A (rubescensin A) and derivant, rubescensine B (rubescensin B), rubescensine C (rubescensin C), rubescensine D (rubescensin D), rubescensine E (rubescensin E), rubescensine H diterpene-kind compound and triterpenoid compound such as oleanolic acid and ursolic acid such as (rubescensin F), and the total terpene content of Rabdosia rubescens is more than 50% in this extract.
2, the described Chinese medicine Rabdosia rubescens of a kind of claim 1 effective site antitumor drug preparation method of extract, it is characterized in that: its technical process may further comprise the steps:
1. pulverize the back Rabdosia rubescens 3-4 time with alcoholic solution or double solvents reflux, extract,, solvent load is 5-8 a times of raw material, obtains the Rabdosia rubescens crude extract;
2. merge crude extract, concentrating under reduced pressure reclaims solvent, is concentrated into relative density 1.05-1.10;
3. with the extract petroleum ether extraction after above-mentioned the concentrating, slough fat and pigment, residue extracted with diethyl ether 3-4 time;
4. merge ethereal extract, reclaim ether, drying gets the total terpene crude extract of Rabdosia rubescens;
5. column chromatographic isolation and purification dissolves the total terpene crude extract of Rabdosia rubescens earlier with low-carbon alcohols, crosses macroporous adsorptive resins, is that mobile phase is carried out eluting with the gradient mixed solvent solution, and reuse TLC method monitoring stream goes out product, collects the outflow product that contains the total terpene of Rabdosia rubescens;
6. merge eluent, concentrating under reduced pressure is removed solvent, and freezing or spray drying is to powder, the total terpene of Rabdosia rubescens, perhaps add the crystallization of hot acetone solution weight again, obtain monomeric compounds such as purer rubescensine A, second element.
3, Chinese medicine Rabdosia rubescens effective site antitumor drug preparation method of extract according to claim 2, it is characterized in that: the leaching process of the total terpene of described Rabdosia rubescens is taked the merceration extraction method, the percolation extraction method, the ultrasonication extraction method, reflux extraction, the extraction solvent that is adopted is an ethanol, ether, acetone or their mixture, described column chromatographic isolation and purification, take with neutrality and polar macroporous adsorption resin, silica gel, aluminium oxide, ion exchange resin is that inserts adsorbs, used eluent can be an ethanol, methanol, chloroform, acetone, petroleum ether or their mixed liquor and aqueous solution, the preferred temperature of described concentrating under reduced pressure is 50-60 ℃, vacuum is 400-600Mpa, and drying condition adopts vacuum drying, the atmospheric steam drying, lyophilization and spray drying.
4, Chinese medicine Rabdosia rubescens effective site antitumor drug preparation method of extract according to claim 2, it is characterized in that: extract is added pharmaceutics acceptable drug adjuvant component, make injection and oral agents, wherein oral formulations comprises capsule, soft capsule, tablet, granule, slow releasing agent, oral liquid etc.; Injection comprises various aqueous injection and injectable powder.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1311814C (en) * | 2004-09-14 | 2007-04-25 | 河南康鑫药业有限公司 | Rabdosia dripping pill and its preparing method |
| CN100370981C (en) * | 2005-11-07 | 2008-02-27 | 上海第二医科大学附属瑞金医院 | Application of Maoeryisu for pharmacy |
| WO2008022505A1 (en) * | 2006-08-18 | 2008-02-28 | Rui Jin Hospital Affiliated To Shanghai Jiao Tong University School Of Medicine | Use of rubescensine a and derivatives thereof in pharmacy |
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| CN1311814C (en) * | 2004-09-14 | 2007-04-25 | 河南康鑫药业有限公司 | Rabdosia dripping pill and its preparing method |
| CN100370981C (en) * | 2005-11-07 | 2008-02-27 | 上海第二医科大学附属瑞金医院 | Application of Maoeryisu for pharmacy |
| CN100402532C (en) * | 2006-02-10 | 2008-07-16 | 陈俊辉 | Preparation method for extracting oridonin from rabdosia |
| WO2008022505A1 (en) * | 2006-08-18 | 2008-02-28 | Rui Jin Hospital Affiliated To Shanghai Jiao Tong University School Of Medicine | Use of rubescensine a and derivatives thereof in pharmacy |
| CN101347500B (en) * | 2007-07-20 | 2011-08-10 | 天津天士力制药股份有限公司 | Effective component of rabdosia and preparation and use thereof |
| CN101874826B (en) * | 2009-10-30 | 2013-04-03 | 湖北省农业科学院农产品加工与核农技术研究所 | Granulose rabdosia rubescens compound preparation and preparation method thereof |
| CN103316076A (en) * | 2013-07-10 | 2013-09-25 | 吴中区胥口精益生物医药研究所 | Esophageal cancer resisting traditional Chinese medicine essence and extraction process thereof |
| CN106053655A (en) * | 2016-06-24 | 2016-10-26 | 广西灵峰药业有限公司 | Quality control method of tablet for improving children appetite |
| CN106619804A (en) * | 2016-09-30 | 2017-05-10 | 河南科技大学 | Herba rabdosiae extract having antibacterial activity as well as preparation method and application of herba rabdosiae extract |
| CN111166752A (en) * | 2020-02-13 | 2020-05-19 | 黑龙江中医药大学 | Pharmaceutical composition for treating cervical cancer and preparation method thereof |
| CN111166752B (en) * | 2020-02-13 | 2021-08-17 | 黑龙江中医药大学 | Pharmaceutical composition for treating cervical cancer and preparation method thereof |
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