CN101554416B - Chinese medicine composition and preparation method and application thereof - Google Patents
Chinese medicine composition and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a Chinese medicine composition and a preparation method and the application thereof. The Chinese medicine composition is prepared by gamboges, agrimony and umbellate pore furgus. The preparation method comprises the following steps: weighting 20-60 of gamboges according to the weight portion, refluxing and extracting by adding ethanol with concentration of 95 percent, and concentrating filter liquor to obtain extract A; weighting 700-1000 agrimony and 500-800 of umbellate pore furgus according to the weight portion, decocting by adding water, and concentrating filter liquor to obtain extract B; drying the extract A and the extract B until the extract A and the extract B become dried paste, crushing the dried paste into fine powder, combining the fine powder, adding excipient, and preparing the preparation according to the common process. The composition can strengthen body resistance and eliminate evil, is safe without obvious side effects, and has remarkable antitumor function.
Description
Technical field
The invention belongs to the Chinese medicine field of traditional Chinese medicine pharmacy, be specifically related to a kind of antineoplastic Chinese medicine composition and preparation method thereof.
Background technology
Malignant tumor is the healthy commonly encountered diseases of serious threat human life, and according to the WHO2000 statistics, there are 1,006 ten thousand tumor new cases in the whole world, and wherein about 6,210,000 people are dead, needs the existing disease patient who treats, looks after to surpass 2,000 ten thousand people.2004 because of malignant tumor death accounts for 24% of Chinese urban and rural residents general mortality rate, first of all kinds of causes of the death of compatriots placed in the middle.
At present, the main means of western medicine treatment malignant tumor are operation, chemotherapy and radiation.In recent years, immunity, endocrine, molecular targeted, laser, method such as photosensitive, freezing also are applied to treat malignant tumor, but the curative effect of the single application of these Therapeutic Method all has than limitations.
The traditional Chinese medical science to tumor treatment be a kind of not only to because of but also suit the medicine to the illness the comprehensive treatment that has not only taken stopgap measures but also effected a permanent cure.Chinese medicine can be adjusted body's immunological function, and reduction is put, chemotherapy toxic side effect, certain tumor-inhibiting anticancer effect is arranged again, thereby obviously improve clinical symptoms, improves life quality, prolongs life cycle, and the patient is taken like a shot.The Chinese medicine worker has proved Chinese medicine from many aspects to the tumor treatment effect.Increase exempt to loose (Radix Ginseng Rubra, the Radix Astragali, Fructus Lycii, Fructus Ligustri Lucidi, Herba Cistanches) like the Gao Shi application Radix Astragali and before 37 routine patient with esophageal carcinoma arts, took for 4 weeks, the histology who observes the excision BIAO and BEN changes.Find that matter mastocyte quantity is assembled with lymphocyte kitchen range shape between the medication group esophageal carcinoma and increase that a matter microvascular lesions is obvious, show that the Radix Astragali increases to exempt from diffusing the patient with esophageal carcinoma immunologic function is had the improvement effect.Sun Shi to be setting upright potentiation side's (Radix Astragali, Caulis Spatholobi, Fructus Lycii, Fructus Ligustri Lucidi, Radix Pseudostellariae, the Rhizoma Atractylodis Macrocephalae, Radix Asparagi) treatment pulmonary carcinoma 16 examples, 3 examples that disappear after the treatment, part 8 examples that disappear, total effective rate 68.8%; Radiation alone 16 examples are respectively 1 example, 4 examples, total effective rate 31.3%.Set upright potentiation side and add the radiotherapy treatment esophageal carcinoma 9 examples, 2 examples that disappear, 5 examples that partly disappear, total effective rate 77.8%.Potentiation is set upright in the experimental studies results prompting can increase the heavy and bone marrow nucleated cell number of mice spleen, and can alleviate the damage of lonizing radiation to macrophage.Wang Shi has observed 158 routine postoperative gastric cancers and has carried out chemotherapy and took right supporting anticancer side's (Radix Astragali, Radix Codonopsis, Semen Coicis, Radix Adenophorae (Radix Glehniae), Radix Ophiopogonis, Rhizoma Pinelliae etc.) simultaneously 1～4 year, survival person 41.2% more than 3 years, more than 5 years more than 29.4%, 10 year 12.5%.Long far beyond the simple chemotherapy person's survival period of postoperative.Experiment detects and to show, this can make the immunoglobulin, E-rosettes, the lymphoblastic transformation rate that are lower than normal level before the treatment recover normal, and can improve the NK cytoactive.Guo Shi uses Radix Arnebiae extract treatment pulmonary carcinoma 19 examples, and the cancer piece person of dwindling 25% forms all and obviously increases than NK cytoactive, IL-2 generation, E-rosettes before the treatment after treating, and carcinoembryonic antigen (CEA) decline.The Wang Shi report cooperates the DVP scheme treatment esophageal carcinoma 437 examples with Rabdosia rubescens, Radix Sophorae Tonkinensis, Herba Solani Nigri mixture, and effect is obvious.Experiment shows that Rabdosia rubescens extract can suppress tumor cell DNA, RNA is synthetic, and it is long-pending to cause that G2 phase and M phase cell increase, so the tumor potentiation of pressing down is arranged.The anti-tumor drink of Xu Shi report lung benefiting (Radix Astragali, Radix Glehniae, Radix Asparagi, Fructus Ligustri Lucidi, Rhizoma Paridis, Herba Selaginellae Doederleinii etc.) has the effect of the lung cancer metastasis of prevention, and its mechanism maybe be with to improve immunologic cellular activity such as NK relevant.Wang Shi finds that with anticancer Bao oral liquid (Radix Astragali, Rhizoma Atractylodis Macrocephalae, Radix Asparagi, Fructus Lycii, Rhizoma Curcumae, Rhizoma Pinelliae (processed with Rhizoma Zingiberis Recens), Herba Hedyotidis Diffusae, Herba Scutellariae Barbatae, Fructus Fici, Fructus Akebiae, Radix Et Rhizoma Rhei, Radix Glycyrrhizae Preparata) treatment late tumor this side has the effect that suppresses tumor growth, prolongation life span, improves patient's life quality.Chen Shi uses the random-effect model of Meta analytical method, to the domestic nineteen ninety～1996 year simple Chinese medicine constitutional nonsmall-cell lung cancer that adopts, and carries out quantitative combined analysis with chemotherapy as the research document of contrast.The result shows: the efficient advantage of chemotherapy is 1.48 times of Chinese medicine, but long term survival rate does not improve.The characteristics of Chinese medicine are the existence of band tumor, and long term survival rate has also shown certain advantage.
Chinese medicine combine with operation, radiotherapy, chemotherapy improve oncotherapy in the recent period and the effect of late result, Chinese medicine can prevent and treat put, chemotherapy toxic side effect.Rational Application Chinese medicine is regulated unbalance interior environment, and it is the foothold of Chinese traditional treatment cancer that organismic internal environment is developed to the direction that is unfavorable for tumor growth.In a word, Chinese medicine cooperates doctor trained in Western medicine method treatment tumor, and the Chinese and western medicine advantage is remedied mutually, maximizes favourable factors and minimizes unfavourable ones, and both can alleviate toxicity, can heighten the effect of a treatment again, improves patient's life quality, prolongs life cycle.The Chinese medicine preparation of effectively treating tumor in the market is few, and the market share is low.Therefore, development pure Chinese medicine anti-tumor agent evident in efficacy, that toxic and side effects is little has significant meaning.
Summary of the invention
The Chinese medicine composition that the purpose of this invention is to provide a kind of evident in efficacy, no obvious toxic-side effects.
Another object of the present invention provides the method for preparing of this Chinese medicine composition.
A further object of the invention provides the application of said composition at the preparation antitumor drug.
A kind of Chinese medicine composition is characterized in that said composition is prepared from following bulk drugs material:
Resina garciniae 20～60 weight portion Herba Agrimoniaes 700～1000 weight portion Polyporus 500～800 weight portions.
Above-mentioned composition is prepared from following bulk drugs material:
Resina garciniae 40 weight portions, Herba Agrimoniae 833 weight portions, Polyporus 667 weight portions.
The dosage form of above-mentioned composition is a said dosage form on any medicament, includes but not limited to granule, tablet, capsule, powder, pill, oral liquid etc.
A kind of method for preparing of Chinese medicine composition is characterized in that may further comprise the steps:
A. take by weighing Resina garciniae 20～60 weight portions and add 95% alcohol reflux three times, merge backflow, filter, filtrating concentrate extractum A;
B. take by weighing Herba Agrimoniae 700～1000 weight portions, Polyporus 500～800 weight portions, decocte with water three times, collecting decoction filters, filtrating concentrate extractum B;
C. extractum A and extractum B are dried to dried cream respectively, are ground into fine powder, merge, add adjuvant, process various dosage forms according to common process.
A kind of method for preparing of Chinese medicine composition is characterized in that specifically can may further comprise the steps:
A. take by weighing Resina garciniae 20～60 weight portions and add 95% alcohol reflux three times, for the first time 6 times of amounts, the second time 3 times of amounts, 3 times of amounts for the third time, each 0.5 hour; Merge backflow; Filter, filtrate recycling ethanol, relative density is 1.25～1.30 extractum A when being evaporated to 50 ℃;
B. take by weighing Herba Agrimoniae 700～1000 weight portions, Polyporus 500～800 weight portions, decocte with water three times, 12 times of amounts for the first time; 10 times of amounts for the second time, 10 times of amounts for the third time, each 2 hours; Collecting decoction filters, and relative density was 1.25～1.30 extractum B when filtrating was concentrated into 50 ℃;
C. extractum A and extractum B are dried to dried cream respectively, are ground into fine powder, merge, add adjuvant, process various dosage forms according to common process.
Drug main of the present invention is wanted strengthening vital QI to eliminate pathogenic factors.Be applicable to before and after treatment operation, the chemicotherapy and should not perform the operation and the malignant tumor patient of chemicotherapy.Chinese medicine composition of the present invention is applied to prepare anti-tumor drug, and safety, no obvious toxic-side effects.
Below come further to set forth the beneficial effect of medicine according to the invention through Test Example, these Test Example have comprised the pharmacodynamic experiment and anxious malicious long term toxication of medicine of the present invention (according to the preparation of embodiment 2 methods, to call the compound agrimony capsule in the following text).
The pharmacodynamic experiment of Test Example 1, medicine of the present invention:
1, the compound agrimony capsule is irritated stomach to mice-transplanted tumor S
180Inhibitory action
Test material
Title: compound agrimony capsule, lot number 20060201; Cyclophosphamide (CTX), specification: the 200mg/ bottle, lot number: 06060121, Hengrui Medicine Co., Ltd., Jiangsu Prov..
Experimental technique:
Get 50 of ICR kind white mice, 18-22g, male and female half and half are pressed transplanted tumor organon inoculation S
180Solid type is inoculated back 24 hours and is claimed that Mus is heavy, and is divided into 5 groups at random, and 10 every group, male and female half and half, blank group and CTX group are respectively the positive and negative matched group.Inoculate administration after 24 hours, the ig administration, administration volume: 0.4ml/20g, once a day, administration is 7 times altogether, and the 2nd day mice weighed after drug withdrawal, puts to death tumor-bearing mice and separates the tumor piece, claims that tumor is heavy, and the gained data are carried out statistical procedures (t check).
The result shows, compares with the blank group, and (300,150mg/kg) group all can suppress animal inhibition sarcoma S to the compound agrimony capsule significantly
180Growth (P＜0.01), less to the influence of the body weight of experiment mice simultaneously.Positive drug CTX group is to S
180The inhibitory action of tumor and to the influence of the body weight of laboratory animal significantly (P＜0.01).The experiment triplicate, the result is close.See table 1.
Table 1 compound agrimony capsule ig is to mice-transplanted tumor S
180Inhibitory action
(n=10)
*P＜0.05
*Compare with the blank group P＜0.01
2, the compound agrimony capsule is irritated the inhibitory action of stomach to mice-transplanted tumor EC
Experimental technique:
Get 50 of above-mentioned specification mices and inoculate back 24 hours and claim that Mus is heavy, and be divided into 5 groups at random by transplanted tumor organon inoculation EC solid type, 10 every group, male and female half and half, blank group and CTX group are respectively the positive and negative matched group.Inoculate administration after 24 hours, the ig administration, administration volume: 0.4ml/20g is total to administration 7 times once a day, and the 2nd day mice weighed after drug withdrawal, puts to death tumor-bearing mice and also separates the tumor piece, claims that tumor is heavy, and the gained data are carried out statistical procedures (t check).
The result shows, compares with the blank group, and (300,150mg/kg) group all can suppress the growth (P＜0.01) that animal suppresses tumor EC to the compound agrimony capsule significantly, and is less to the body weight influence of experiment mice simultaneously.Positive drug CTX group is to the inhibitory action of EC tumor with to the body weight influence of laboratory animal significantly (P＜0.01).The experiment triplicate, the result is close.See table 2.
Table 2 compound agrimony capsule ig to the inhibitory action
of mice-transplanted tumor EC (n=10)
*P＜0.05
*Compare with the blank group P＜0.01
3, the compound agrimony capsule is irritated the inhibitory action of stomach to mice-transplanted tumor Heps
Experimental technique:
Get 50 of above-mentioned specification mices and inoculate back 24 hours and claim that Mus is heavy, and be divided into 5 groups at random by transplanted tumor organon inoculation Heps solid type, 10 every group, male and female half and half, blank group and CTX group are respectively the positive and negative matched group.Inoculate administration after 24 hours, the ig administration, administration volume: 0.4ml/20g is total to administration 7 times once a day, and the 2nd day mice weighed after drug withdrawal, puts to death tumor-bearing mice and also separates the tumor piece, claims that tumor is heavy, and the gained data are carried out statistical procedures (t check).
The result shows, compares with the blank group, and (300,150mg/kg) group all can suppress the growth (P＜0.01) that animal suppresses hepatocarcinoma tumor Heps to the compound agrimony capsule significantly, and is less to the body weight influence of experiment mice simultaneously.Positive drug CTX group is to the inhibitory action of Heps tumor with to the body weight influence of laboratory animal significantly (P＜0.01).The experiment triplicate, the result is close.See table 3.
Table 3 compound agrimony capsule ig to the inhibitory action
of mice-transplanted tumor Heps (n=10)
*P＜0.05
*Compare with the blank group P＜0.01
Conclusion:
The compound agrimony capsule (300,150,75mg/kg) irritate stomach (ig) administration to mice transplanted tumor S180; EC, the growth of Heps all has the obvious suppression effect, and wherein compound agrimony capsule (300mg/kg) ig is to mice transplanted tumor S180; EC; The growth inhibition ratio best result Bie Keda 53.4%, 55.98% and 56.62% of Heps, less to the body weight influence of experiment mice simultaneously.But CTX is to transplanted tumor S180 under the similarity condition, EC, the inhibitory action of Heps and to the body weight influence of laboratory animal significantly.
Test Example 2 compound agrimony capsules are to the effect of people's Gastric Cancer MGC-803 bare mouse different species transplantation tumor growth inhibited
Test material:
The compound agrimony capsule, lot number: 20060201;
Topotecan，5.7mg/ml。Producer: Yangzhou Olympic Competition health pharmaceutcal corporation, Ltd.Lot number: 20060101 transplanted tumoies, people's Gastric Cancer MGC-803 Nude Mice, it is subcutaneous and set up to be inoculated in the nude mouse axillary fossa by people's Gastric Cancer MGC-803 cell strain.The cell inoculation amount is 1 * 10
6, inoculation is used after forming and in the nude mouse body, passing for 3 generations again after the transplanted tumor.
Experimental technique:
The tumor tissue of getting the growth animated period cuts into 1.5mm
3About, under aseptic condition, be inoculated in the BALB/c nude mouse (35-40 days; 18-22g; Female) the right side axillary fossa is subcutaneous.Nude Mice treats that with vernier caliper measurement transplanted tumor diameter tumor growth is to 100mm
3After with the animal random packet, 8 every group.Use the method for measuring the tumor footpath, dynamic observe by examination thing antineoplastic effect.The measurement number of times of diameter of tumor is 3 times weekly, claims simultaneously when measuring that Mus is heavy at every turn.The compound agrimony capsule on every Mondays, three, five gastric infusions 3 times, the administration volume is 0.2ml/20g.The Topotecan matched group is intravenously administrable 2 times weekly, and negative control group is irritated stomach and given the equivalent normal saline solution.After 4 weeks of administration, mice is put to death, and operation strips the tumor piece and weighs.Gross tumor volume (tumor volume, computing formula TV) is:
TV=1/2 * a * b
2(wherein a, b represent length and width respectively)
The compound agrimony capsule is seen table 4 to the experimental therapeutic outcome of people's Gastric Cancer MGC-803 Nude Mice, table 5 and Fig. 1, Fig. 2.Experimental result shows that the compound agrimony capsule has certain inhibitory action to the growth of people's Gastric Cancer MGC-803 Nude Mice.When compound agrimony capsule gastric infusion 300mg/kg, 150mg/kg, 75mg/kg dosage, the tumour inhibiting rate best result of people's Gastric Cancer MGC-803 Nude Mice is not reached 55.2% and 46.7%, 29.1%.Tumour inhibiting rate to people's Gastric Cancer MGC-803 Nude Mice when Topotecan is with the 2mg/kg intravenous injection under the similarity condition is 65.7%.The capsular antitumor action of compound agrimony is certain dose-effect relationship.
Table 5. compound agrimony capsule is to the inhibitory action of people's Gastric Cancer MGC-803 bare mouse different species transplantation tumor growth
(X±SD，n＝8)
Compare with blank control group,
*P＜0.05,
*P＜0.01
The compound agrimony capsule shows that to the experimental therapeutic outcome of people's Gastric Cancer MGC-803 Nude Mice the compound agrimony capsule has certain inhibitory action to the growth of people's Gastric Cancer MGC-803 Nude Mice.When compound agrimony capsule gastric infusion 300mg/kg, 150mg/kg, 75mg/kg dosage, the tumour inhibiting rate best result of people's Gastric Cancer MGC-803 Nude Mice is not reached 55.2% and 46.7%, 29.1%.Tumour inhibiting rate to people's Gastric Cancer MGC-803 Nude Mice when Topotecan is with the 2mg/kg intravenous injection under the similarity condition is 65.7%.
Test Example 3, medicine acute toxicity testing of the present invention
Experiment material:
1, compound agrimony capsule, every gram extract powder are equivalent to 9.49 gram raw medicinal herbs, lot number: 20060201.
2,0.5% sodium carboxymethyl cellulose (CMC-Na) solution, preparation in advance regularly, cold preservation (2-8 ℃) vial is preserved, and is used to prepare compound agrimony capsule suspension.
Experimental animal:
40 of healthy ICR mices, cleaning level, male and female half and half, body weight 18-22g; 40 of healthy SD rats, cleaning level, male and female half and half, body weight 130-150g.By the supply of Zhejiang Province's Experimental Animal Center, the animal quality quality certification: SCXK (Zhejiang) 2003-0001.
Dosage group and definite foundation:
According to mice, rat acute toxicity prerun result and toxic reaction situation, the complete dead dosage of the anxious malicious prerun oral administered compound XIANHECAO JIAONANG of mice is 10.0g/kg, and the dosage of living entirely is 6.0g/kg; The complete dead dosage of the anxious malicious prerun oral administered compound XIANHECAO JIAONANG of rat is 10.0g/kg, and the dosage of living entirely is 4.5g/kg.Mice respectively is provided with following dosage group with group apart from 0.8 with group distance 0.84, rat.
Mice:
The 1st group of compound agrimony capsule 6.0g/kg
The 2nd group of compound agrimony capsule 7.1g/kg
The 3rd group of compound agrimony capsule 8.4g/kg
The 4th group of compound agrimony capsule 10.0g/kg
Rat:
The 1st group of compound agrimony capsule 5.1g/kg
The 2nd group of compound agrimony capsule 6.4g/kg
The 3rd group of compound agrimony capsule 8.0g/kg
The 4th group of compound agrimony capsule 10.0g/kg
Above dose groups is faced the time spent and is mixed with the compound agrimony capsule suspension of variable concentrations, equal capacity with 0.5% sodium carboxymethyl cellulose (CMC-Na) solution, supplies mice, rat oral gavage to use.Administration capacity: mice: 0.8ml/20g body weight; Rat: 2.0ml/100g body weight.
Test method:
Get 40 of ICR mices, 40 of SD rats are divided into above each dose groups at random, and 10 every group, male and female half and half.Fasting can't help water more than 12 hours (beginning fasting ten eight: 30 evening before that day, 9 preceding administrations in morning next day) before each dose groups administration.Irritating stomach gives above each dose groups compound agrimony capsule on an empty stomach respectively; At once observe the response situation of animal after the administration; Comprise that animal appearance, behavioral activity, the mental status, appetite, defecation and color thereof, fur, nose, eye, mouth have or not abnormal secretion thing and death condition; The timely postmortem of dead animal is if perusal has obvious pathological changes internal organs then to carry out histopathologic examination.And administration the 1st day every at a distance from 30 minutes to 1 h observation 1 time, observe 6h continuously, observe later every day 1 time, observed continuously 14 days, the record animal toxicity reacts and dead distribution situation.Handle with the NDST statistical software according to mortality rate, calculate mice, the oral acute half lethal dose (LD of rat by the Bliss method
50) and 95% credible limit value, tabulation expression.
Result of the test:
1. mice difference single oral (po) (consistent with the clinical administration approach) gives compound agrimony capsule 6.0,7.1,8.4,10.0g/kg dosage; Abnormal response does not all appear in administration each group at once; Medicine appearance loose stool, urinary incontinence (dripping urine), movable digestion, urinary system and psychologic nervous system reaction such as reduce, close one's eyes appear in administration after 2～3 hours; 24 hours and 48 hours are observed respectively after the administration, and 6.0,7.1, each survival mice of 8.4g/kg dose groups progressively recovers normal activity.Administration begins to occur dead after 23 hours, each was organized mortality rate and was respectively 0%, 30%, 70%, 100% after the administration between 23 hours to 72 hours the death time.Its reaction occurrence rate, extent of reaction, duration of the reaction, dead occurrence rate and dead time of occurrence all are corresponding relation with dosage, and the high reaction of dosage weighs, time of occurrence morning, longer duration, mortality rate are also high, and dosage hangs down then opposite.Handle with the NDST statistical software, calculate the mice single oral by the Bliss method and give compound agrimony capsular acute half lethal dose (LD
50) and 95% credible limit value be 7.7257 (7.2403～8.2437) g/kg.Dead animal pathological anatomy macroscopy is not seen except that the gastric drug residue obviously unusually, and off-test is cutd open all survival mice of inspection and also do not seen obvious macroscopic internal organs pathological change.
2. rat difference single oral (po) gives compound agrimony capsule 5.1,6.4,8.0,10.0g/kg dosage; At once respectively organize after the administration and abnormal response all do not occur; Digestion, urinary system and psychiatric system reactions such as sialorrhea, medicine appearance loose stool, urinary incontinence (dripping urine), movable minimizing, lethargy appear in administration after 30 minutes～1 hour; Perpendicular hair, reaction such as hunchbacked, prostrate appear in administration after 2 hours; The oral cavity nasal cavity appearred in 6 hours later in administration has color secretions phenomenon, and administration is each dose groups rat phenomenon (especially 10.0g/kg dosage is obvious) that all occurs becoming thin after the 2nd day.5.1g/kg after the dose groups administration 48 hours, 6.4,72 hours above-mentioned toxic reactions of each survival rats fade away after the administration of 8.0g/kg dose groups.Administration begins to occur dead after 8 hours, each was organized mortality rate and was respectively 10%, 30%, 80%, 90% after the administration between 8 hours to 72 hours the death time.Its reaction occurrence rate, extent of reaction, duration of the reaction, mortality rate and dead time of occurrence all are corresponding relation with dosage, and the high reaction of dosage weighs, time of occurrence morning, longer duration, mortality rate are also high, and dosage hangs down then opposite.Handle with the NDST statistical software, calculate the acute half lethal dose (LD of rat oral administered compound XIANHECAO JIAONANG by the Bliss method
50) and 95% credible limit value be 6.9931 (6.2545～7.819) g/kg.Dead animal pathological anatomy macroscopy is not seen except that the gastric drug residue obviously unusually, and off-test is cutd open all survival rats of inspection and do not seen obvious macroscopic internal organs pathological change.
Test Example 4, medicine long term toxicity test of the present invention
Test material:
1, compound agrimony capsule, every gram extract powder are equivalent to 9.49 gram raw medicinal herbs, and lot number 20060201 is provided by China Medicine University's development.
2,0.5% sodium carboxymethyl cellulose (CMC-Na) solution, preparation in advance regularly, cold preservation (2-8 ℃) vial is preserved, and is used to prepare compound agrimony capsule suspension.
Experimental animal:
The SD rat, cleaning level, age 5-6 age in week, body weight: ♂ 86～104g, ± s:94.9 ± 4.6; ♀ 85～103g, ± s:94.6 ± 4.8; Totally 120, each 60 of ♀ ♂.By Zhejiang Province's Experimental Animal Center supply, the animal quality quality certification number: SCXK (Zhejiang) 2003-0001.
Dosage divides into groups:
High dose group compound agrimony capsule 1.0g/ (kgd) * 180d
Middle dose groups compound agrimony capsule 0.5g/ (kgd) * 180d
Low dose group compound agrimony capsule 0.25g/ (kgd) * 180d
Matched group 0.5%CMC-Na solution 10ml/ (kgd) * 180d
More than each group face with preceding and be mixed with variable concentrations, the equal suspension of capacity with 0.5%CMC-Na solution, confession rat oral gavage usefulness, the administration capacity is the 1.0ml/100g body weight, it is identical that each organizes the administration capacity.
Dosage is confirmed foundation:
According to the clinical plan of people with dosage 1.953g/ people d (press the 60kg body weight and calculate, be i.e. 0.03g/kgd);
2. be 6.0g/kg according to acute toxicity trial test mice single oral (po) administration dosage alive entirely, rat single oral (po) administration dosage alive entirely is 4.5g/kg.
Test method:
Get 120 of cleaning level SD rats, numbering, male be odd number, female is even number, is divided into four test group by randomized blocks, i.e. high, medium and low three dose groups of compound agrimony capsule and a matched group, 30 every group, respectively 15 of male and female.Observe a week before the test, the behavioral activity of record rat, ingest, drink water, spirit, hair, body weight etc.Duration of test, per os of same time of every morning is irritated stomach (po) and is administered once (consistent with the clinical administration approach), and administration is 6 days weekly, continuous 180 days (6 months).Weigh weekly once, with body weight increase and decrease adjustment dosage.Every morning the observed and recorded rat general signs (comprising behavior, motor function, breathing, hair color, mouth, eye, nose, ear, excrement, urine etc.), food ration, amount of drinking water and death condition; Discovery has the animal of toxic reaction to take out single cage raising; Primary part observation; Discovery has dead or the timely postmortem of dying animal, does to see substantially and histopathologic examination.
Hematology, blood biochemical, organ weights, coefficient and pathomorphology and histological examination are carried out in administration mid-term (administration three months), drug withdrawal next day (administration six months) and drug withdrawal one month (convalescent period end) respectively.Fasting can't help water more than 12 hours (preceding a whole night of 20 beginnings fasting, blood sampling about 8: 30 morning of next day) before the inspection, behind etherization, and ventral aorta blood sampling, EDTA-K
2Anticoagulated whole blood is measured erythrocyte (RBC), leukocyte (WBC), haemachrome (HGB), platelet (PLT), packed cell volume (HCT), MCVU (MCV), average hemoglobin (MCH), NCHC (MCHC), erythrocyte volume distribution density (RDW), mean platelet volume (MPV), MPW (PDW), thrombocytocrit (PCT) and lymphocyte (Lym#), intermediate value cell (Mid#) and granulocyte (Grn#) with Sha jasmine Automatic Blood Cell Analyzer.Measure prothrombin time (PT) with the CA-100 coagulo meter.Measure reticulocyte (Ret) with brilliant cresyl blue test tube staining.Serum is measured aspartic acid transferring enzyme (AST), alanine aminotransferase (ALT), alkali phosphatase (ALP), blood urea nitrogen (BUN), creatinine (Crea), total protein (T.P), albumin (ALB), blood glucose (GLU), STB (T.BIL), T-CHOL (T.CHO) with HITACHI7020 automatic clinical chemistry analyzer and pairing reagent; Creatine phosphokinase (CK), triglyceride (TG) are measured Na with NOVA 10 electrolyte appearance and pairing reagent
+, K
+, Cl
-, TCa concentration.Administration mid-term, drug withdrawal next day and convalescent period finish to put to death 1/3 rat respectively and do system's postmortem and histopathologic examination, measure the organ weights and the coefficient (%) of the heart, liver, spleen, lung, kidney, brain, thymus, adrenal gland, thyroid, testis, epididymis, prostate, uterus, ovary; Core simultaneously, liver, spleen, lung, kidney, adrenal gland, thyroid, pancreas, stomach, duodenum, ileum, colon, hypophysis, brain, spinal cord, breastbone, thymus, lymph node, bladder, uterus, ovary, the attached testis of testis, prostate make paraffin section, HE dyeing, do pathology histological examination, observation and analysis with Nikon PF microscope and histopathology image analysis system.Above each item observation index (initial data) all carries out statistical analysis and processing with NDST software kit and the corresponding statistical procedure of EXCEL; With the basic data that statistical significance, general response situation, the result of histopathologic examination combine this laboratory to set up, overall merit result of the test.
Result of the test:
Continuous six months of overview SD rat respectively per os give compound agrimony capsule 1.0,0.5 and 0.25g/ (kgd) dosage, only high dose group part animal appearance activity reduce, abnormal responses such as perpendicular hair, depilation, hair are fluffy, medicine appearance loose stool, dripping urine.3 animals of the 33rd day～the 153rd day high dose group of administration occur dead midway, wherein 2 heros (149
#, 179
#) 1 female (356
#), high dose group animal dead rate is 10%.In, any abnormal response all do not appear in all rats of low dose group during whole test, do not cause animal dead yet, and is identical with control rats.
Continuous six months of body weight SD rat respectively per os give compound agrimony capsule 1.0,0.5 and 0.25g/ (kgd) dosage, mainly the high dose group rat body weight is increased and has a significant effect.Be embodied in: the female rats of high dose group was tested for the 10th, 12,14,16,20～22 weeks (the 9th, 11,13,15,19～21 weeks of administration), and male rat was tested for the 3rd, 6,7,10～27 weeks the average weight of (administration the 2nd, 5,6,9～26); The average weight that middle dose groups male rat was tested for the 14th week (the 13rd week of administration) is lighter than the matched group (P＜0.05,0.01) of corresponding time.Middle dose groups female rats tested for the 3rd, 4 weeks, and the low dose group male rat is tested the matched group (P＜0.05) that the 4th～6 all average weights overweight the corresponding time.
3. ingest with drink water continuous six months of SD rat respectively per os give compound agrimony capsule 1.0,0.5 and 0.25g/ (kgd) dosage, to the ingesting, drink water not to have basically obviously and influence of rat, the irregular increase and decrease of drinking-water that indivedual cycles only occur is fluctuateed.Be embodied in the male Mus of high, medium and low dose groups and tested for the 17th week (the 16th week of administration), the male Mus of middle dose groups was tested for the 20th week the matched group (P＜0.05) of the average amount of drinking water in (the 19th week of administration) more than the corresponding time; And the female Mus of low dose group was tested for the 21st week the matched group (P＜0.01) that the average amount of drinking water in (the 20th week of administration) is less than the corresponding time.Other times respectively organize rat amount of drinking water and food ration all similar (P＞0.05) with the matched group of corresponding time.
Continuous six months of hematology SD rat respectively per os give compound agrimony capsule 1.0,0.5 and 0.25g/ (kgd) dosage, rat blood is learned index does not have obvious influence.Only high dose group RBC, HGB, HCT, PLT, PCT, RDW%, MCV, the PT in administration mid-term, the RBC of middle dose groups, HCT, PT, RDW%, the HGB of low dose group, HCT, PT meansigma methods are higher than the matched group (P＜0.05,0.01) of corresponding time.And MCH, the MCHC in high dose group administration mid-term, Lym#, WBC, MPV, the Mid# of drug withdrawal next day, Lym#, WBC that convalescent period finishes; In MCH, MCHC, the MPV in dose groups administration mid-term, the Lym# of drug withdrawal next day, Lym#, MPV that convalescent period finishes; The MPV meansigma methods that low dose group administration mid-term and convalescent period finish is lower than the matched group (P＜0.05,0.01) of corresponding time.But more than there is the measured value of index its each Mus except that Lym# of height difference to belong to the range of normal value that this laboratory is set up basically, and do not have tangible dose-response relationship.Other indexs then similar with matched group (P＞0.05) all belong to range of normal value.
Blood biochemical learn continuous six months of SD rat respectively per os give compound agrimony capsule 1.0,0.5 and 0.25g/ (kgd) dosage, mainly part blood biochemical high, middle dose groups rat is learned index (Cl
-↑, CK ↓) certain influence arranged.Be embodied in and finish high dose group drug withdrawal next day, convalescent period, middle dose groups administration mid-term, drug withdrawal next day, convalescent period finish, the Cl in low dose group administration mid-term
-Value is higher than the matched group (P＜0.05,0.01) of corresponding time; And high dose group administration mid-term, drug withdrawal next day and convalescent period finish and in dose groups drug withdrawal next day and convalescent period the CK value that finishes be lower than the matched group (P＜0.05,0.01) of corresponding time; The indivedual biochemical indicators of high, middle dose groups (AST, GLU, T.P, T.CHO, BUN, Na
+, K
+And TCa) though with the matched group of corresponding time significant difference (P＜0.05,0.01) is arranged, it changes irregularities, and the measured value of these its each Mus of index still belongs to the range of normal value that this laboratory is set up.Other indexs then similar with matched group (P＞0.05) all belong to range of normal value.
Continuous six months of organ weights and coefficient S D rat respectively per os give compound agrimony capsule 1.0,0.5 and 0.25g/ (kgd) dosage, organ weights and the coefficient of rat do not had obvious influence.The kidney organ coefficient of the liver organ coefficient of only high, middle dose groups administration mid-term and drug withdrawal next day, high dose drug withdrawal next day, the adrenal gland's organ weights of high dose group drug withdrawal next day and the meansigma methods of coefficient are higher than the matched group (P＜0.05,0.01) of corresponding time; Other indivedual internal organs (ovary, testis, epididymis, thyroid, prostate) are though weight and the coefficient of indivedual time periods have significant difference (P＜0.05,0.01) with matched group; But it changes irregularities, and it is seen substantially, and all discovery is obviously unusual with histopathologic examination.The weight of other internal organs and coefficient then similar (P＞0.05) with the matched group of corresponding time.
7. see substantially and continuous six months of histopathologic examination SD rat respectively per os give compound agrimony capsule 1.0,0.5 and 0.25g/ (kgd) dosage, thymus, spleen, lymph node, liver, stomach, intestinal, the bone marrow of the dead rat of high dose group is had certain damaging action; Liver, small intestinal to high dose group biopsy rat have the reversibility damaging action.Be embodied in: 3 dead midway rats of high dose group during the administration; Wherein 2 rats are owing to mainly organize internal organs self-dissolving to occur and can't carry out histopathologic examination, and 1 (No. 356) dead rat outward appearance is become thin in addition, analyse see subcutaneous and internal organs between fatty tissue rare; Intestinal wall is thin; The bilateral adrenal gland is red and swollen, atrophy of thymus gland, and the lung color and luster is scarlet; Histological examination mainly shows as thymus, spleen, lymph node, bone marrow atrophy, hepatocyte atrophy, smaller volume, stomach, intestinal shallow-layer mucosal degeneration, the slight atrophy of body of gland.Administration mid-term and drug withdrawal time daily inspection, the figure is all thinner for high dose group biopsy rat outward appearance; The hepatocyte cloudy swelling all appears in histological examination, and administration has 5 high dose group rats (5/9) mucous membrane of small intestine epithelial cell vacuolar degeneration mid-term, but the similar variation of small intestinal is not seen in drug withdrawal next day; Convalescent period finishes inspection high dose group rat and respectively organizes internal organs not see that all obviously unusual pathology change.In each inspection phase, low dose group and control rats respectively organize internal organs not see that all obviously unusual substantive pathology change; Only show as the cytopathy of little kitchen range property of indivedual rat hearts, liver, prostate, kidney appearance; Stroma cell, lymphocyte slightly soak into; Degree is slight and do not have the dosage dependency relation, is thought of as non-medicine factor affecting.
Above result of the test shows; Under this test dose condition; Continuous six months of SD rat respectively per os to give lot number be 20060201 compound agrimony capsule 1.0,0.5 and 0.25g/ (kgd) dosage; Toxicities such as its main toxicity is the minimizing of high dose group part activities in rats, medicine appearance loose stool, dripping urine, perpendicular hair, depilation, body weight gain is slow or alleviate are mainly liver, intestinal, stomach, thymus, spleen, lymph node and bone marrow to the poisoning target organ of dead rat during the administration; Poisoning target organ to the biopsy rat is liver, small intestinal, and belongs to reversibility influence (analogue is not seen in a drug withdrawal inspection in month).The dosage of being poisoned to death is 1.0g/ (kgd), and safe dose is 0.5g/ (kgd), and non-toxic is 0.25g/ (kgd).
Description of drawings
Fig. 1 is the influence sketch map of compound agrimony capsule to people's Gastric Cancer MGC-803 bare mouse different species transplantation tumor growth change in volume.
Fig. 2 is that the compound agrimony capsule is to people's Gastric Cancer MGC-803 bare mouse different species transplantation tumor growth inhibited effect sketch map.
The specific embodiment
The preparation of embodiment 1 present composition granule
A. take by weighing the 20g Resina garciniae and add 95% alcohol reflux three times, for the first time 6 times of amounts, 3 times of amounts, 3 times of amounts for the third time for the second time, each 0.5 hour, the merging backflow, filtration, filtrate recycling ethanol, relative density is the extractum A of 1.25-1.30 when being evaporated to 50 ℃.
B. take by weighing Herba Agrimoniae 1000g, Polyporus 800g, decocte with water three times, 12 times of amounts for the first time, 10 times of amounts for the second time, 10 times of amounts for the third time, each 2 hours, collecting decoction filtered, and relative density is the extractum B of 1.25-1.30 during filtrating vacuum concentration to 50 ℃.
C. with extractum A and extractum B difference vacuum drying (≤60 ℃) to dried cream, be ground into fine powder (100 order) back and merge, adding starch is an amount of, and mix homogeneously is processed soft material in right amount with 70% ethanol, granulation, and≤60 ℃ of dryings, granulate promptly gets 1000 bags of granules.
The preparation of embodiment 2 present composition capsules
A. take by weighing the 40g Resina garciniae and add 95% alcohol reflux three times, for the first time 6 times of amounts, 3 times of amounts, 3 times of amounts for the third time for the second time, each 0.5 hour, the merging backflow, filtration, filtrate recycling ethanol, relative density is the extractum A of 1.25-1.30 when being evaporated to 50 ℃.
B. take by weighing Herba Agrimoniae 833g, Polyporus 667g, decocte with water three times, 12 times of amounts for the first time, 10 times of amounts for the second time, 10 times of amounts for the third time, each 2 hours, collecting decoction filtered, and relative density is the extractum B of 1.25-1.30 during filtrating vacuum concentration to 50 ℃.
C. with extractum A and extractum B difference vacuum drying (≤60 ℃) to dried cream, be ground into fine powder (100 order) back and merge, adding starch is an amount of; Mix homogeneously is processed soft material in right amount with 70% ethanol, and 18 order stainless steel sifts are granulated; ≤60 ℃ of dryings, granulate adds 0.5% magnesium stearate; Mix homogeneously is filled 1000 capsules.
The preparation of embodiment 3 present composition tablets
A. take by weighing the 60g Resina garciniae and add 95% alcohol reflux three times, for the first time 6 times of amounts, 3 times of amounts, 3 times of amounts for the third time for the second time, each 0.5 hour, the merging backflow, filtration, filtrate recycling ethanol, relative density is the extractum A of 1.25-1.30 when being evaporated to 50 ℃.
B. take by weighing Herba Agrimoniae 700g, Polyporus 500g, decocte with water three times, 12 times of amounts for the first time, 10 times of amounts for the second time, 10 times of amounts for the third time, each 2 hours, collecting decoction filtered, and relative density is the extractum B of 1.25-1.30 during filtrating vacuum concentration to 50 ℃.
C. with extractum A and extractum B difference vacuum drying (≤60 ℃) to dried cream, be ground into fine powder (100 order) back and merge, adding starch is an amount of, and mix homogeneously is processed soft material in right amount with 70% ethanol, granulation, and≤60 ℃ of dryings are pressed 1000 of tablet common process compactings.
The preparation of embodiment 4 present composition pills
A. take by weighing the 50g Resina garciniae and add 95% alcohol reflux three times, for the first time 6 times of amounts, 3 times of amounts, 3 times of amounts for the third time for the second time, each 0.5 hour, the merging backflow, filtration, filtrate recycling ethanol, relative density is the extractum A of 1.25-1.30 when being evaporated to 50 ℃.
B. take by weighing Herba Agrimoniae 900g, Polyporus 750g, decocte with water three times, 12 times of amounts for the first time, 10 times of amounts for the second time, 10 times of amounts for the third time, each 2 hours, collecting decoction filtered, and relative density is the extractum B of 1.25-1.30 during filtrating vacuum concentration to 50 ℃.
C. with extractum A and extractum B difference vacuum drying (≤60 ℃) to dried cream, be ground into fine powder (100 order) back and merge, add adjuvant, process 1000 pills.
The preparation of embodiment 5 present composition oral liquids
A. take by weighing the 30g Resina garciniae and add 95% alcohol reflux three times, for the first time 6 times of amounts, 3 times of amounts, 3 times of amounts for the third time for the second time, each 0.5 hour, the merging backflow, filtration, filtrate recycling ethanol, relative density is the extractum A of 1.25-1.30 when being evaporated to 50 ℃.
B. take by weighing Herba Agrimoniae 750g, Polyporus 600g, decocte with water three times, 12 times of amounts for the first time, 10 times of amounts for the second time, 10 times of amounts for the third time, each 2 hours, collecting decoction filtered, and relative density is the extractum B of 1.25-1.30 during filtrating vacuum concentration to 50 ℃.
C. with extractum A and extractum B vacuum drying (≤60 ℃) to dried cream respectively, merge after being ground into fine powder, process the 1000ml oral liquid by the oral liquid common process.
Claims (5)
1. Chinese medicine composition is characterized in that said composition is prepared from following bulk drugs:
Resina garciniae 20～60 weight portion Herba Agrimoniaes 700～1000 weight portion Polyporus 500～800 weight portions,
The above-mentioned raw materials medicine is prepared Chinese medicine composition according to the following steps:
A. take by weighing Resina garciniae 20～60 weight portions and add 95% alcohol reflux three times, merge backflow, filter, filtrating concentrate extractum A;
B. take by weighing Herba Agrimoniae 700～1000 weight portions, Polyporus 500～800 weight portions, decocte with water three times, collecting decoction filters, filtrating concentrate extractum B;
C. extractum A and extractum B are dried to dried cream respectively, are ground into fine powder, merge, add adjuvant, process required dosage form according to common process.
2. Chinese medicine composition according to claim 1 is characterized in that said composition is prepared from following bulk drugs material:
Resina garciniae 40 weight portions, Herba Agrimoniae 833 weight portions, Polyporus 667 weight portions.
3. Chinese medicine composition according to claim 1 and 2 is characterized in that described dosage form is to add conventional adjuvant, according to common process, processes granule, tablet, capsule, powder, pill, the oral liquid of clinical acceptance.
4. the method for preparing of the described Chinese medicine composition of claim 1 is characterized in that this method may further comprise the steps:
A. take by weighing Resina garciniae 20～60 weight portions and add 95% alcohol reflux three times, merge backflow, filter, filtrating concentrate extractum A;
B. take by weighing Herba Agrimoniae 700～1000 weight portions, Polyporus 500～800 weight portions, decocte with water three times, collecting decoction filters, filtrating concentrate extractum B;
C. extractum A and extractum B are dried to dried cream respectively, are ground into fine powder, merge, add adjuvant, process required dosage form according to common process.
5. the application of the described Chinese medicine composition of claim 1 in the preparation antitumor drug.
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|---|---|---|---|---|
| CN1208638A (en) * | 1997-08-15 | 1999-02-24 | 毕盛 | Medicine for treating cancer and method for preparing same |
| CN1507884A (en) * | 2002-12-16 | 2004-06-30 | 辉 张 | Chinese medicine powder and decoction composition for treating cancers |
| CN101115493A (en) * | 2005-08-12 | 2008-01-30 | 河乃建仁 | Agent for hair growth |
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2008
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1208638A (en) * | 1997-08-15 | 1999-02-24 | 毕盛 | Medicine for treating cancer and method for preparing same |
| CN1507884A (en) * | 2002-12-16 | 2004-06-30 | 辉 张 | Chinese medicine powder and decoction composition for treating cancers |
| CN101115493A (en) * | 2005-08-12 | 2008-01-30 | 河乃建仁 | Agent for hair growth |
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| 曹迪，等.中医药治疗肝癌的研究进展.《中华现代临床医学杂志》.2004,第2卷(第10期), * |
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