CN102145158A - Ginger and clove composition, preparation method of ginger and clove composition and use of ginger and clove composition for preparing toxicity-reducing efficacy-improving medicine in cancer radiotherapy and chemotherapy - Google Patents
Ginger and clove composition, preparation method of ginger and clove composition and use of ginger and clove composition for preparing toxicity-reducing efficacy-improving medicine in cancer radiotherapy and chemotherapy Download PDFInfo
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Abstract
The invention relates to a natural pharmaceutical composition, a preparation method of the natural pharmaceutical composition and the use of the natural pharmaceutical composition for preparing toxicity-reducing efficacy-improving medicine in cancer radiotherapy and chemotherapy. The natural pharmaceutical composition consists of decocted and concentrated extract of ginger and clove, gingerol, clove ethanol extract and eugenol based on the proportion of 3 to 6: 2 to 5: 3 to 5: 2 to 9, and the composition has extremely good therapeutic effect for vomiting and other side effects caused by cancer radiotherapy and chemotherapy and can simultaneously play the role of inhibiting tumor.
Description
Technical field
The present invention relates to a kind of natural drug composition, relate more specifically to a kind of natural drug composition of being made up of Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol, the preparation method of this natural drug composition and this natural drug composition are being used for preparing the purposes that reduces the vomiting side effect that cancer is put, chemotherapy causes and the treatment cancer is played the medicine of potentiation.
Background technology
Cancer has become the big killer that the mankind face at present, and mortality rate is only second to cardiovascular disease, and is the trend that rises year by year.There is 180 extremely evil property tumor patients every year in China, wherein 2/3 patient accept to put, chemotherapy, though certain curative effect is arranged, put, in various degree side effect can appear in most of patient after the chemotherapy, wherein modal side effect be feel sick, gastrointestinal reaction such as vomiting.Frequent vomiting can cause Digestive system to be lost in a large number, and water, electrolyte, acid base imbalance make patient agonizing, have a strong impact on patient's life quality, not only can have influence on curative effect, even makes patient be difficult to stand and therapy discontinued.
At present, clinical Bendectin commonly used mainly contains histamine (H
2), acetylcholine (M) and dopamine (DA) receptor antagonist and cannabis medicine etc., but the weak curative effect of these medicines, and toxic and side effects is strong, is very limited in the application to putting, vomitting after the chemotherapy.Novel Bendectin is as NK
1Receptor antagonist and 5-HT
3Receptor antagonist class medicine, though to put, the vomiting antiemetic effect is good after the chemotherapy, costs an arm and a leg, the scope of application is narrow, and toxicity is big, clinically needs drug combinations more, and does not see the anticancer synergia effect, the development of the natural anti-cancer attenuation synergistic medicine that therefore waits in expectation.
Chinese medicine for cancer put, the research of chemotherapy attenuation synergistic, attract people's attention.Present still unmatchful anticancer disease is put, chemotherapy has the Chinese patent medicine that alleviates vomiting side effect and potentiation.Therefore, research and develop a kind of high-efficiency low-toxicity, cheap, can be used for of being easy to promote alleviate that cancer is put, emesis of chemotherapy side effect and the natural Chinese medicine that plays the efficacy enhancing and toxicity reducing effect become clinical problem demanding prompt solution.
Chinese medicine Rhizoma Zingiberis Recens (zingiber) is described as " vomiting panacea ".Gingerol is the active ingredient that extracts from Rhizoma Zingiberis Recens, the inventor observe gingerol can resist inorganic and organic poison copper sulfate and cisplatin due to vomit, and suppress the intestinal pheochromocyte and discharge to cause and tell factor 5-HT and P material, gingerol acts on the maximally related NK of emesis of chemotherapy
1And 5-HT
3Deng receptor, be many target spots, natural low toxicity Bendectin.Flos Caryophylli is alabastrum and the fruit of Myrtaceae syzygium species plant Flos Caryophylli Eugenia caryophyllata Thunb.The energy warming the stomach, the kidney warming.Be used for the treatment of singultus, vomiting and diarrhoea.In recent years, Rhizoma Zingiberis Recens and Flos Caryophylli antitumaous effect are confirmed, especially Rhizoma Zingiberis Recens piquancy component and eugenol, the inventor is through research and the experiment and the clinical practice of each a large amount of formula proportion, obtained optimal drug ratio compositions of the present invention at last, be the compositions of the novelty of agreeing with mutually with polyceptor target spot ratio, and proved that it has fabulous curative effect to the side effect such as vomiting that cancer is put, chemotherapy causes and can play auxiliary effect that suppresses tumor simultaneously, has finished the present invention thus.
Summary of the invention
First purpose of the present invention is to provide a kind of natural drug composition, and the said composition low toxicity is efficient, determined curative effect, is convenient to use and promotes, both can antitumor, and can resist again and side effect such as put, vomit after the chemotherapy.
Second purpose of the present invention is to provide the preparation method of above-mentioned natural drug composition.
The 3rd purpose of the present invention is to provide that above-mentioned natural drug composition reduces in preparation that cancer is put, chemotherapy side effect and the treatment cancer is played purposes in the medicine of efficacy enhancing and toxicity reducing effect.
A kind of natural drug composition provided by the invention, by Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol by weight in 3-6: the ratio of 2-5: 3-5: 2-9 is formed.
Above-mentioned natural drug composition, wherein the usage ratio of Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol is 6: 5: 5: 9.
Wherein, Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction can be by well known to a person skilled in the art the conventional method acquisition of purifying, gingerol and eugenol even can commerce buy adopt these compositions that conventional method obtains or the natural drug of the present invention that adopts the commercial one-tenth of buying to be grouped into all can obtain identical curative effect.The method of below listing only is that the inventor thinks preferred method, and is not intended to the extracting method of each composition in this natural drug composition is made special restriction.
Preferably, Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration preparation method of extract is in the above-mentioned natural drug composition:
According to proportion compatibility is 1: 1 or 1-4: 1 other ratios are got Rhizoma Zingiberis Recens, Flos Caryophylli, 10 times of amount distilled water immersion 1h, heating decocts 1h on the electric furnace that adds asbestos gauge, and silk filters, preserve filtrate, medicinal residues add 6 times of distilled water again, soak, decoct, filter with method, and twice filtrate is mixed, heating is concentrated into small size, ultrafiltration membrance filter adopts 70 ℃ of drying under reduced pressure 36h of vacuum drying oven to get pale brown toner end, is Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract.
The preparation method of gingerol is in the above-mentioned natural drug composition:
Rhizoma Zingiberis Recens is thinly sliced, and natural drying breaks into powder with the high speed Universalpulverizer, crosses 30 mesh sieves, and the fine powder that will sieve is again put into the big port grinding bottle of 5000mL, soaks 72h down with 6 times of amount acetone room temperature; At 40 ℃, supersound extraction 15min under the ultrasonic frequency of 40KHz, the centrifugal supernatant that obtains, residue discards; With supernatant decompress filter again, filtrate places the Rotary Evaporators vacuum concentration to get oil of ginger shape mixture, and carrying out this mixture repeatedly again, the concentrate drying purification obtains gingerol.
Flos Caryophylli ethanol extraction preparation method is in the above-mentioned natural drug composition:
Get Flos Caryophylli, drying is pulverized, sieve,, cross the leaching clear liquor with 7-10 times of soak with ethanol 12h, residue continues to soak 12h, and twice filtrate is merged centrifugal (3000rpm) 15 minutes, separation of supernatant, ethanol and impurity are removed in distillation, add ethyl acetate extraction, drying under reduced pressure becomes dry extract again, pulverize, cross 100 mesh sieves, promptly.
The eugenol preparation method is in the above-mentioned natural drug composition:
Flos Caryophylli is pulverized, with the little backflow 30min down that boils of 95% ethanol, filter, filtrate is fixed (pH=9～10) with 5% sodium hydroxide, filters, and steams and removes ethanol, uses the 25mL extracted with diethyl ether 3 times at every turn; The water layer hcl acidifying is transferred pH value of solution=5, uses extracted with diethyl ether 3 times, and combining extraction liquid is used anhydrous sodium sulfate drying, steams and removes ether, gets the pale yellow oily liquid body, is eugenol.
The present invention also provides the preferred manufacturing procedure of this natural drug composition, comprising:
(1) preparation of Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract
According to proportion compatibility is 1: 1 or 1-4: 1 other ratios are got Rhizoma Zingiberis Recens, Flos Caryophylli, 10 times of amount distilled water immersion 1h, heating decocts 1h on the electric furnace that adds asbestos gauge, and silk filters, preserve filtrate, medicinal residues add 6 times of distilled water again, soak, decoct, filter with method, and twice filtrate is mixed, heating is concentrated into small size, ultrafiltration membrance filter adopts 70 ℃ of drying under reduced pressure 36h of vacuum drying oven to get pale brown toner end, is Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract.
(2) preparation of gingerol
Rhizoma Zingiberis Recens is thinly sliced, and natural drying breaks into powder with the high speed Universalpulverizer, crosses 30 mesh sieves, and the fine powder that will sieve is again put into the big port grinding bottle of 5000mL, soaks 72h down with 6 times of amount acetone room temperature; At 40 ℃, supersound extraction 15min under the ultrasonic frequency of 40KHz; The centrifugal supernatant that obtains, residue discards; With supernatant decompress filter again, filtrate places the Rotary Evaporators vacuum concentration to get oil of ginger shape mixture, and carrying out this mixture repeatedly again, the concentrate drying purification obtains gingerol.
(3) Flos Caryophylli ethanol extraction preparation
Get Flos Caryophylli, drying is pulverized, sieve,, cross the leaching clear liquor with 7-10 times of soak with ethanol 12h, residue continues to soak 12h, and twice filtrate is merged centrifugal (3000rpm) 15 minutes, separation of supernatant, ethanol and impurity are removed in distillation, add ethyl acetate extraction, drying under reduced pressure becomes dry extract again, pulverize, cross 100 mesh sieves, promptly.
(4) eugenol preparation
Flos Caryophylli is pulverized, with the little backflow 30min down that boils of 95% ethanol, filter, filtrate is fixed (pH=9～10) with 5% sodium hydroxide, filters, and steams and removes ethanol, uses the 25mL extracted with diethyl ether 3 times at every turn; The water layer hcl acidifying is transferred pH value of solution=5, uses extracted with diethyl ether 3 times, and combining extraction liquid is used anhydrous sodium sulfate drying, steams and removes ether, gets the pale yellow oily liquid body, is eugenol.
(5) choose Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol in proportion, uniform mixing, promptly.
The preparation method of above-mentioned natural drug composition, also comprise: the product that step (5) makes is further made the capsule that contains natural drug composition 100mg or 200mg specification by the pharmacy means of routine, maybe will make the tablet that contains natural drug composition 100mg or 200mg specification by wet granulation technology.
The present invention comprises also that natural drug composition reduces in preparation that cancer is put, chemotherapy side effect and the treatment cancer is played purposes in the medicine of potentiation, and reduces the vomiting reaction that cancer is put, chemotherapy causes and the treatment cancer is played purposes in the medicine of potentiation in preparation.
The instructions of taking of natural drug composition of the present invention: the treatment cancer puts, available capsule of vomiting or tablet after the chemotherapy, and 600mg divides three times one after each meal for each person every day.
The beneficial effect of natural drug composition of the present invention aspect resisting emesis and inhibition tumor is as follows:
1, natural drug composition of the present invention has good resisting emesis effect, has now finished clinical preceding pharmaceutical research at mink vomiting model.The mink vomiting model that adopts us to initiate is observed this natural drug composition cisplatin is caused the antagonism over a period to come of mink vomiting model.Studies show that: this natural drug composition has tangible antagonism for the vomiting reaction of mink due to the cisplatin, and its effect is better than using separately Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction.Simultaneously, its reduction causes the rate aspect of vomitting, and its effect matches in excellence or beauty with the Western medicine ondansetron.
2, natural drug composition of the present invention has the effect that suppresses tumor, has now finished clinical preceding pharmaceutical research on mouse tumor model.Adopted mouse tumor model, observed the inhibitory action of natural drug composition mouse interior tumor.Studies show that: this natural drug composition has the obvious suppression effect for mouse interior tumor.
3, proved that in addition this natural drug composition can strengthen the inhibitory action of chemotherapeutics to tumor, now finished clinical preceding pharmaceutical research on the mouse tumor cell model.Adopted mouse tumor model, observed this natural drug composition and cyclophosphamide combined inhibitory action mouse interior tumor.Studies show that: this natural drug composition can obviously increase the inhibitory action of cyclophosphamide for tumor.
4, finished simultaneously in the research of the acute toxicology of mice, studies show that: this natural drug composition does not have influence to feed and behavioral activity etc., and internal organs and pathological observation be no abnormality seen also.Illustrate that this natural drug composition toxicity is very little.
5, the inventor makes up Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol originally, and by the consumption proportion between definite these components of a large amount of experiments, make said composition bring into play good synergism, obtain unexpected effect.Pass through our experimentation repeatedly, make the inventor surprised be ratio between each component of natural drug composition of the present invention at 3-6: 2-5: 3-5: produced unexpected synergism during 2-9, potentiation is obvious, extraordinary effect is played in side effect, especially the usage ratio in each component is 6: 5: 5: 9 o'clock especially obvious, this is that the inventor pays a large amount of creative works by keeping punching and obtains on the paper basis of delivering before, and those of ordinary skills only can't simply obtain this ratio range and optimum amount ratio by disclosed content.
6, this natural drug composition Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and each component of eugenol all can obtain by conventional means, some of them component for example gingerol and eugenol also can be buied by commerce, and such compositions itself has and obtains convenient, lower-cost effect.This also is the product that has splendid curative effect and higher industrial practicality concurrently that the inventor obtains by a large amount of creative work sides.
The specific embodiment
Further set forth the preparation of natural drug composition of the present invention below by embodiment and test example, with and the beneficial effect in resisting emesis and antitumor, used.
Embodiment
The first step: the preparation of various extracts
(1) preparation of Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract
According to proportion compatibility is 1: 1 or 1-4: 1 other ratios are got Rhizoma Zingiberis Recens, Flos Caryophylli, 10 times of amount distilled water immersion 1h, heating decocts 1h on the electric furnace that adds asbestos gauge, and silk filters, preserve filtrate, medicinal residues add 6 times of distilled water again, soak, decoct, filter with method, and twice filtrate is mixed, heating is concentrated into small size, ultrafiltration membrance filter adopts 70 ℃ of drying under reduced pressure 36h of vacuum drying oven to get pale brown toner end, is Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract.
(2) preparation of gingerol
Rhizoma Zingiberis Recens is thinly sliced, and natural drying breaks into powder with the high speed Universalpulverizer, crosses 30 mesh sieves, and the fine powder that will sieve is again put into the big port grinding bottle of 5000mL, soaks 72h down with 6 times of amount acetone room temperature; At 40 ℃, supersound extraction 15min under the ultrasonic frequency of 40KHz; The centrifugal supernatant that obtains, residue discards; With supernatant decompress filter again, filtrate places the Rotary Evaporators vacuum concentration to get oil of ginger shape mixture, and carrying out this mixture repeatedly again, the concentrate drying purification obtains gingerol.
(3) Flos Caryophylli ethanol extraction preparation
Get Flos Caryophylli, drying is pulverized, sieve,, cross the leaching clear liquor with 7-10 times of soak with ethanol 12h, residue continues to soak 12h, and twice filtrate is merged centrifugal (3000rpm) 15 minutes, separation of supernatant, ethanol and impurity are removed in distillation, add ethyl acetate extraction, drying under reduced pressure becomes dry extract again, pulverize, cross 100 mesh sieves, promptly.
(4) eugenol preparation
Flos Caryophylli is pulverized, with the little backflow 30min down that boils of 95% ethanol, filter, filtrate is fixed (pH=9～10) with 5% sodium hydroxide, filters, and steams and removes ethanol, uses the 25mL extracted with diethyl ether 3 times at every turn; The water layer hcl acidifying is transferred pH value of solution=5, uses extracted with diethyl ether 3 times, and combining extraction liquid is used anhydrous sodium sulfate drying, steams and removes ether, gets the pale yellow oily liquid body, is eugenol.
Second step: the preparation of preparation
(1) preparation of natural drug composition capsule preparations of the present invention
Take by weighing Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract 24mg, gingerol 20mg, Flos Caryophylli ethanol extraction 20mg and the eugenol 36mg of above-mentioned steps preparation respectively, take by weighing starch 395mg, magnesium stearate 5mg simultaneously, uniform mixing, filled capsules is made the capsule of each pure heavy 500mg.
(2) preparation of natural drug composition tablet of the present invention
Take by weighing Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract 24mg, gingerol 20mg, Flos Caryophylli ethanol extraction 20mg and the eugenol 36mg of above-mentioned steps preparation respectively, take by weighing starch 295mg, lactose 100mg simultaneously, uniform mixing, with wet granulation, drying, mix with the 5mg magnesium stearate behind the granulate, tabletting is made the tablet of every pure heavy 500mg.
Drug effect and pharmacology's data:
Compositions is in following test example: the usage ratio of Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol is 6: 5: 5: 9.The inventor to above-mentioned each composition usage ratio at 3-6: 2-5: 3-5: the compositions that the multiple proportioning combination in the time of between the 2-9 obtains has been carried out and last identical experiment, find that all the natural drug composition in this amount ranges can obtain good synergism, to cancer put, chemotherapy side effect can play good effect and treatment for cancer is had potentiation.And, the inventor finds that Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol can also be the compositions that obtains with conventional means, and be not only the composition that obtains with above-mentioned method, the compositions that above-mentioned four kinds of one-tenth of buying that made by conventional means or commercial are grouped into has the identical effect of compositions that makes with said method, its experimental data is no longer enumerated here, and those skilled in the art can approve this.Below be drug effect and the pharmacology's data that preferred embodiment obtains:
Test example 1: the pharmacodynamic study of natural drug composition treatment vomiting effect
1, test material
(1) Flos Caryophylli is bought by the big pharmacy of Qingdao all living creatures, and Rhizoma Zingiberis Recens is purchased Yushan Hill degree of conquering east, and is accredited as the alabastrum of Myrtaceae syzygium species plant Flos Caryophylli and the tuber of zingiber by the director of studio of this institute.
(2) cisplatin (cisplatin), Shandong Qilu Pharmaceutical Factory produces, lot number: 903005CE, 20mg/ props up.
(3) Ondansetron Hydrochloride injection, the pharmacy of Olympic Competition health, lot number: 090102,2ml:4mg.
(4) healthy adult mink, body weight 1.3-1.8kg, male, provide by special cultivation animal feeding center, Qingdao, every animal places the ferrum cage of 75cm * 50cm * 50cm separately, 24h natural light irradiation, ad lib drinking-water before the experiment.
2, test method:
Mink is divided into 7 groups at random, every group 6, the A group is the gingerol group, the B group is Flos Caryophylli ethanol extraction group, the C group is gingerol and Flos Caryophylli ethanol extraction compatibility group (ratio is 1: 1), and the D group is Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, and (Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol were by 6: 5: 5: 9 mixed) for natural drug composition of the present invention for the E group, the F group is the cisplatin positive controls, and the G group is the ondansetron group.The A group is earlier with gingerol 10g.kg
-1(being equivalent to contain the crude drug amount) ig pretreatment, 30min gives cisplatin 7.5mg.kg
-1Ip; The B group gives Flos Caryophylli ethanol extraction 10g.kg
-1(being equivalent to contain the crude drug amount) ig pretreatment, 30min gives cisplatin 7.5mg.kg
-1Ip; The C group gives gingerol and Flos Caryophylli ethanol extraction compatibe drug 10g.kg
-1(being equivalent to contain the crude drug amount) ig pretreatment, 30min gives cisplatin 7.5mg.kg
-1Ip; The D group gives Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract 10g.kg
-1(being equivalent to contain the crude drug amount) ig pretreatment, 30min gives cisplatin 7.5mg.kg
-1Ip; The E group gives natural drug composition 10g.kg of the present invention
-1(being equivalent to contain the crude drug amount) ig pretreatment, 30min gives cisplatin 7.5mg.kg
-1Ip; The F group gives normal saline, and 30min gives cisplatin 7.5mg.kg
-1Ip; The G group is earlier with ondansetron group 7.5g.kg
-1Pretreatment, 30min gives cisplatin 7.5mg.kg
-1Ip.For observing mink vomiting reaction in the 6h after administration, write down its vomiting incubation period, retch and vomiting number of times.
3, experimental result
Compare with matched group, each treatment group all can obviously alleviate retch and vomiting number of times and prolong vomits incubation period (P＜0.01).Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract wherein, natural drug composition group of the present invention prolong vomiting and are better than the ondansetron group incubation period.Gingerol group, gingerol and Flos Caryophylli ethanol extraction compatibility group, Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract group, and natural drug composition of the present invention can significantly reduce the rate of vomitting that causes.Wherein Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract group is identical with the ondansetron group; And causing the rate of vomitting, natural drug composition group of the present invention is lower than the ondansetron group.The results are shown in Table 1.
(7.5mg/kg ip) causes the influence of vomitting and acting on to table 1 extract compatibility to cisplatin
Annotate: a is equivalent to contain the crude drug amount, compares with matched group
cP＜0.01
Test example 2: the pharmacodynamic study of natural drug composition antitumor action of the present invention
1, experiment material
(1) Kunming mouse, body weight 20 ± 2g is provided by Qingdao City medicine inspecting institute Experimental Animal Center.Ad lib drinking-water before the experiment, the 24h natural light irradiation.
(2) cyclophosphamide, the permanent auspicious medical Pharmacy stock Co., Ltd in Jiangsu product.
2, test method:
(1) to mice S
180The inhibition test of sarcoma growth
Employing was gone down to posterity 5-7 days, and well-grown milky ascites is diluted to tumor cell suspension with an amount of physiological saline solution, and cell number is 10
7/ mL, 12 every group, the right axillary fossa subcutaneous vaccination of every Mus 0.2mL (contains oncocyte 2 * 10
6Individual).Inoculation random packet next day, experimental group natural drug composition of the present invention is divided into (400,200,100mg/mL) three dosage groups, every Mus is irritated stomach 0.5ml; Negative control group gives 0.5mL distilled water; The positive drug group gives cyclophosphamide 20gkg
-1D
-1, successive administration 10 days.The conventional raising, drinking-water, food is not limit.Drug withdrawal next day, mice all takes off cervical vertebra puts to death, and strips subcutaneous solid tumor tumor piece, weighs, and calculates tumor control rate (%) by following formula:
Tumor control rate=(the average tumor of the matched group average tumor of heavy (C)-experimental group heavy (T))/average tumor of matched group heavy (C)
(2) to rat liver cancer H
22The inhibition test of growth
Grouping, dosage, administering mode, experimental period and S
180The sarcoma strain is identical.
(3) to the inhibition test of mice ehrlich ascites tumor (solid tumor)
Grouping, dosage, administering mode, experimental period and S
180The sarcoma strain is identical.
3, result of the test
Three dosage groups of natural drug composition of the present invention are to S
180, H
22, EAC and negative control group relatively, inhibitory action is in various degree all arranged, especially the highest to the suppression ratio of tumor with high dose group.(as table 2,3,4)
Table 2 capsule is to S
180The tumor inhibition effect of mice
Compare with negative control group,
bP＜0.05,
cP＜0.01
Table 3 capsule is to H
22The tumor inhibition effect of mice
Compare with negative control group,
bP＜0.05,
cP＜0.01
Table 4 capsule is to the tumor inhibition effect of EAC mice
Compare with negative control group,
cP＜0.01
Test example 3: natural drug composition of the present invention is for the pharmacodynamic study of chemotherapy medicine antitumor potentiation
1, experiment material
(1) Kunming mouse, body weight 20 ± 2g is provided by Qingdao City medicine inspecting institute Experimental Animal Center.Ad lib drinking-water before the experiment, the 24h natural light irradiation.
(2) cyclophosphamide, the permanent auspicious medical Pharmacy stock Co., Ltd in Jiangsu product.
2, test method:
(1) to mice S
180The inhibition test of sarcoma growth
Employing was gone down to posterity 5-7 days, and well-grown milky ascites is diluted to tumor cell suspension with an amount of physiological saline solution, and cell number is 10
7/ mL, 12 every group, the right axillary fossa subcutaneous vaccination of every Mus 0.2ml (contains oncocyte 2 * 10
6Individual).The inoculation next day random packet, experimental group natural drug composition of the present invention is divided into (10,5,2.5gkg
-1) three dosage groups, every Mus is irritated stomach 0.5ml, gives cyclophosphamide 10gkg simultaneously
-1D
-1Negative control group gives 0.5ml distilled water; The positive drug group gives cyclophosphamide 20gkg
-1D
-1, successive administration 10 days.The conventional raising, drinking-water, food is not limit.Drug withdrawal next day, mice all takes off cervical vertebra puts to death, and strips subcutaneous solid tumor tumor piece, weighs, and calculates tumor control rate (%) by following formula:
Tumor control rate=(the average tumor of the matched group average tumor of heavy (C)-experimental group heavy (T))/average tumor of matched group heavy (C)
(2) to rat liver cancer H
22The inhibition test of growth
Grouping, dosage, administering mode, experimental period and S
180The sarcoma strain is identical.
(3) to the inhibition test of mice ehrlich ascites tumor (solid tumor)
Grouping, dosage, administering mode, experimental period and S
180The sarcoma strain is identical.
3, result of the test
Under the situation that reduces the cyclophosphamide dosage, three dosage of natural drug composition of the present invention and cyclophosphamide combined medication group are to S
180, H
22, EAC and negative control group relatively, inhibitory action is in various degree all arranged, especially the highest to the suppression ratio of tumor with high dose group.(as table 5,6,7)
Table 5 capsule is to S
180The tumor inhibition effect of mice
Compare with negative control group,
bP＜0.05,
cP＜0.01
Compatibility drug group height: cyclophosphamide 10gkg
-1+ compositions 10gkg
-1In the compatibility drug group: cyclophosphamide 10gkg
-1+ compositions 5gkg
-1The compatibility drug group is low: cyclophosphamide 10gkg
-1+ compositions 2.5gkg
-1, down together
Table 6 capsule is to H
22The tumor inhibition effect of mice
Compare with negative control group,
bP＜0.05
Table 7 capsule is to the tumor inhibition effect of EAC mice
Compare with negative control group,
bP＜0.05,
cP＜0.01
Test example 4: the acute toxicity test of natural drug composition of the present invention
1, experimental animal:
Kunming mouse, body weight 20 ± 2g is provided by Qingdao City medicine inspecting institute Experimental Animal Center
2, medicine:
Natural drug composition of the present invention, tragakanta
3, test method:
According to " the study of tcm new drug specification requirement " of National Drug Administration's promulgation in one on November 12, in 999 revision, natural drug of the present invention is carried out acute toxicity testing.Because of being subjected to drug level and volume restrictions, can't measure LD
50So, carry out the maximum dosage-feeding experiment.Get 30 of Kunming mouses, female half and half, ig gives capsule Chinese medicine 1g/mL, every 0.8mL, and it is long-pending that promptly the maximum of mice is irritated body of stomach, observes mice active state, diet, feces, breathing, body weight and death condition after the medication, observes 14d. continuously
4, result of the test:
Each organizes well-grown after the mouse stomach administration, weight increase, and behavior is normal, there is no death and significant reaction.Behind sacrifice of animal, the no abnormal phenomenon of each main organs of perusal.
The result shows that natural drug composition maximal dose oral application of the present invention does not have obvious damage to animal, does not find the toxic reaction to the body generation, is a kind of safe and reliable medicine.
Claims (7)
1. natural drug composition, it is characterized in that said composition by Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol by weight in 3-6: the ratio of 2-5: 3-5: 2-9 is formed,
Wherein Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration preparation method of extract is:
According to proportion compatibility is 1: 1 or 1-4: 1 other ratios are got Rhizoma Zingiberis Recens, Flos Caryophylli, 10 times of amount distilled water immersion 1h, heating decocts 1h on the electric furnace that adds asbestos gauge, and silk filters, preserve filtrate, medicinal residues add 6 times of distilled water again, soak, decoct, filter with method, and twice filtrate is mixed, heating is concentrated into small size, ultrafiltration membrance filter adopts 70 ℃ of drying under reduced pressure 36h of vacuum drying oven to get pale brown toner end, is Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract.
Wherein the preparation method of Flos Caryophylli ethanol extraction is:
Get Flos Caryophylli, drying is pulverized, sieve,, cross the leaching clear liquor with 7-10 times of soak with ethanol 12h, residue continues to soak 12h, and twice filtrate is merged centrifugal (3000rpm) 15 minutes, separation of supernatant, ethanol and impurity are removed in distillation, add ethyl acetate extraction, drying under reduced pressure becomes dry extract again, pulverize, cross 100 mesh sieves, promptly.
Wherein the preparation method of eugenol is:
Flos Caryophylli is pulverized, with the little backflow 30min down that boils of 95% ethanol, filter, filtrate is fixed (pH=9～10) with 5% sodium hydroxide, filters, and steams and removes ethanol, uses the 25mL extracted with diethyl ether 3 times at every turn; The water layer hcl acidifying is transferred pH value of solution=5, uses extracted with diethyl ether 3 times, and combining extraction liquid is used anhydrous sodium sulfate drying, steams and removes ether, gets the pale yellow oily liquid body, is eugenol.
2. natural drug composition as claimed in claim 1, wherein the usage ratio of Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol is 6: 5: 5: 9.
3. natural drug composition as claimed in claim 1, wherein the preparation method of gingerol is:
Rhizoma Zingiberis Recens is thinly sliced, and natural drying breaks into powder with the high speed Universalpulverizer, crosses 30 mesh sieves, and the fine powder that will sieve is again put into the big port grinding bottle of 5000mL, to soak 72h under the alcohol at normal temperature of 6 times of amounts 95%; At 40 ℃, supersound extraction 15min under the ultrasonic frequency of 40KHz, the centrifugal supernatant that obtains, residue discards; With supernatant decompress filter again, filtrate places the Rotary Evaporators vacuum concentration to get oil of ginger shape mixture, and carrying out this mixture repeatedly again, the concentrate drying purification obtains gingerol.
4. as the preparation method of the natural drug composition of claim 1 or 2, it is characterized in that this method comprises:
(1) preparation of Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract
According to proportion compatibility is 1: 1 or 1-4: 1 other ratios are got Rhizoma Zingiberis Recens, Flos Caryophylli, 10 times of amount distilled water immersion 1h, heating decocts 1h on the electric furnace that adds asbestos gauge, and silk filters, preserve filtrate, medicinal residues add 6 times of distilled water again, soak, decoct, filter with method, and twice filtrate is mixed, heating is concentrated into small size, ultrafiltration membrance filter adopts 70 ℃ of drying under reduced pressure 36h of vacuum drying oven to get pale brown toner end, is Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract.
(2) preparation of gingerol
Rhizoma Zingiberis Recens is thinly sliced, and natural drying breaks into powder with the high speed Universalpulverizer, crosses 30 mesh sieves, and the fine powder that will sieve is again put into the big port grinding bottle of 5000mL, to soak 72h under the alcohol at normal temperature of 6 times of amounts 95%; At 40 ℃, supersound extraction 15min under the ultrasonic frequency of 40KHz, the centrifugal supernatant that obtains, residue discards; With supernatant decompress filter again, filtrate places the Rotary Evaporators vacuum concentration to get oil of ginger shape mixture, and carrying out this mixture repeatedly again, the concentrate drying purification obtains gingerol.
(3) Flos Caryophylli ethanol extraction preparation
Get Flos Caryophylli, drying is pulverized, sieve,, cross the leaching clear liquor with 7-10 times of soak with ethanol 12h, residue continues to soak 12h, and twice filtrate is merged centrifugal (3000rpm) 15 minutes, separation of supernatant, ethanol and impurity are removed in distillation, add ethyl acetate extraction, drying under reduced pressure becomes dry extract again, pulverize, cross 100 mesh sieves, promptly.
(4) eugenol preparation
Flos Caryophylli is pulverized, with the little backflow 30min down that boils of 95% ethanol, filter, filtrate is fixed (pH=9～10) with 5% sodium hydroxide, filters, and steams and removes ethanol, uses the 25mL extracted with diethyl ether 3 times at every turn; The water layer hcl acidifying is transferred pH value of solution=5, uses extracted with diethyl ether 3 times, and combining extraction liquid is used anhydrous sodium sulfate drying, steams and removes ether, gets the pale yellow oily liquid body, is eugenol.
(5) choose Rhizoma Zingiberis Recens Flos Caryophylli water boiling concentration extract, gingerol, Flos Caryophylli ethanol extraction and eugenol uniform mixing in proportion, promptly.
5. as the preparation method of claims 4 described natural drug compositions, also comprise: the product that step (5) makes is further made the capsule that contains natural drug composition 100mg or 200mg specification by the pharmacy means of routine, maybe will make the tablet that contains natural drug composition 100mg or 200mg specification by wet granulation technology.
As each described natural drug composition of claim 1-3 reduce in preparation that cancer is put, chemotherapy side effect and the treatment cancer is played purposes in the medicine of potentiation.
7. as the purposes of each described natural drug composition of claim 1-3 in preparing the medicine that reduces the vomiting reaction that cancer is put, chemotherapy causes and the treatment cancer is played potentiation.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1111149A (en) * | 1995-02-22 | 1995-11-08 | 胡青松 | Dizzy-proof food and its additive |
| CN101066438A (en) * | 2007-06-04 | 2007-11-07 | 济南康众医药科技开发有限公司 | Hiccup treating medicine and its prepn process |
-
2011
- 2011-03-31 CN CN2011100794498A patent/CN102145158B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1111149A (en) * | 1995-02-22 | 1995-11-08 | 胡青松 | Dizzy-proof food and its additive |
| CN101066438A (en) * | 2007-06-04 | 2007-11-07 | 济南康众医药科技开发有限公司 | Hiccup treating medicine and its prepn process |
Non-Patent Citations (4)
| Title |
|---|
| 《云南中医学院学报》 19870331 戴云华等 挥发油与中草药（二） 39-41 1-7 第10卷, 第1期 * |
| 《云南中医学院学报》 19870331 戴云华等 挥发油与中草药（二） 39-41 1-7 第10卷, 第1期 2 * |
| 《泰山医学院学报》 20110131 于一等 白豆蔻汤和丁香汤对顺铂所致水貂呕吐模型的治疗作用 26-27 1-7 第32卷, 第1期 * |
| 《泰山医学院学报》 20110131 于一等 白豆蔻汤和丁香汤对顺铂所致水貂呕吐模型的治疗作用 26-27 1-7 第32卷, 第1期 2 * |
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| CN103263475A (en) * | 2013-04-16 | 2013-08-28 | 阚兆云 | Traditional chinese medicine composition |
| CN103316191A (en) * | 2013-07-11 | 2013-09-25 | 石强 | Natural medicine extract |
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