CN102133384A - Ginger and dried orange peel composition, preparation method thereof and application of ginger and dried orange peel composition in preparation of attenuated synergistic medicaments in cancer radiotherapy and chemotherapy - Google Patents
Ginger and dried orange peel composition, preparation method thereof and application of ginger and dried orange peel composition in preparation of attenuated synergistic medicaments in cancer radiotherapy and chemotherapy Download PDFInfo
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Abstract
The invention relates to a natural pharmaceutical composition, a preparation method thereof and an application of the natural pharmaceutical composition in preparation of attenuated synergistic medicaments in cancer radiotherapy and chemotherapy. The natural pharmaceutical composition is prepared from extract obtained by decocting and concentrating ginger and dried orange peel decoction, gingerol, dried orange peel water extract and nobiletin according to the ratios of (3-5):(2-9):(1-6):(2-5). The composition has an excellent effect on treating emesis and other side effects caused by cancer radiotherapy and chemotherapy, and can simultaneously play a role in inhibiting tumors.
Description
Technical field
The present invention relates to a kind of natural drug composition, relate more specifically to a kind ofly fry in shallow oil the natural drug composition that concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin are formed by Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup, 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.Pericarpium Citri Reticulatae, the mature peel for rutaceae orange (Citrusreticulata Blanco) and variety thereof has another name called Pericarpium Citri tangerinae, have spleen invigorating and regulate the flow of vital energy, the effect of drying dampness to eliminate phlegm, can and in, can control stomach-QI being unable to descend normally, nausea and vomiting.Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is one of China's prescription commonly used, has the warming middle-JIAO to arrest vomiting effect.This side is made up of Rhizoma Zingiberis Recens and Pericarpium Citri Reticulatae, and prescription is simple, and cost is very low.In recent years, Rhizoma Zingiberis Recens and Pericarpium Citri Reticulatae especially Pericarpium Citri Reticulatae alkaloid antitumaous effect are confirmed, and (nobiletin NOB) claims the Fructus Citri tangerinae flavone again to Nobiletin, is to extract from Fructus Aurantii Immaturus and a kind of polymethoxyflavone class material of obtaining.Chemical constitution: 5,6,7,8,3', 4'-hexamethoxy flavone.Foreign study shows in recent years: Nobiletin has pharmacotoxicological effect widely, receives much attention with antitumor action especially, and it is confirmed the curative effect of gastroenteric tumor, cerebroma, colon cancer, breast carcinoma, skin carcinoma and myeloid leukemia.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, it is the compositions of the novelty of agreeing with mutually with polyceptor target spot ratio, and proved that it has fabulous curative effect and can play the effect that suppresses tumor simultaneously the side effect such as vomiting that cancer is put, chemotherapy causes, 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, fry in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin by weight in 3-5 by Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup: the ratio of 2-9: 1-6: 2-5 is formed.
Above-mentioned natural drug composition, wherein to fry in shallow oil the usage ratio of concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin be 5: 9: 6 to Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup: 5.
Wherein, Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin can be by well known to a person skilled in the art the conventional method acquisition of purifying, gingerol and Nobiletin can commerce be buied, and the natural drug of the present invention that adopts these compositions of conventional method acquisition or adopt 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, the preparation method that Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil concentrated extract in the above-mentioned natural drug composition is:
According to proportion compatibility is to get Rhizoma Zingiberis Recens, Pericarpium Citri Reticulatae at 1: 1, adds 10 times of amount distilled water, soaks 1h, heating decocts 1h on the electric furnace that adds asbestos gauge, silk filters, and preserves filtrate, and medicinal residues add 6 times of distilled water again, soak, decoct, filter with method, twice filtrate is mixed, and heating is concentrated into small size, ultrafiltration membrance filter, adopt 70 ℃ of drying under reduced pressure 36h of vacuum drying oven to get yellow powder, be Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup and fry in shallow oil concentrated 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.
Pericarpium Citri Reticulatae water extract preparation method is in the above-mentioned natural drug composition:
Get the Pericarpium Citri Reticulatae medical material, after the coarse pulverization, sieve, decoct with water 3 times, each 2 hours.The water that adds for the first time 10 times of amounts filters, and medicinal residues add the water of 8 times of amounts for the second time, filters, and adds the water of 6 times of amounts for the third time, filters.3 filtrates are merged, centrifugal (3000rpm) 15 minutes, separation of supernatant, water bath method concentrates, and drying under reduced pressure becomes dry extract again, pulverizes, and crosses 100 mesh sieves, promptly.
The Nobiletin preparation method is in the above-mentioned natural drug composition:
Get the Pericarpium Citri Reticulatae medical material, coarse pulverization is sieved, and measures the petroleum ether reflux, extract, 5 times for 20 times, each 6h merges petroleum ether extract, behind the recovery petroleum ether, with the mixed solvent heating for dissolving of 2 kinds of solvent compositions of 0.25 times of amount, place, to be crystallized separating out fully, after the filtration, dry under 80 ℃ the Pericarpium Citri Reticulatae total flavones.Measure through the HPLC method, the content of main effective ingredient Nobiletin is 58.4% in the Pericarpium Citri Reticulatae total flavones, and Pericarpium Citri Reticulatae total flavones ethyl acetate extraction concentrates, the utilization polyamide column chromatography, with the mixed solvent eluting of 2 kinds of solvent compositions,, collect and contain the Nobiletin eluent and volatilize solvent in conjunction with thin layer chromatography, recrystallizing methanol, get yellow solid, fusing point 137-138 ℃, be Nobiletin.
The present invention also provides the preferred manufacturing procedure of this natural drug composition, comprising:
(1) Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil the preparation of concentrated extract
According to proportion compatibility is to get Rhizoma Zingiberis Recens, Pericarpium Citri Reticulatae at 1: 1, adds 10 times of amount distilled water, soaks 1h, heating decocts 1h on the electric furnace that adds asbestos gauge, silk filters, and preserves filtrate, and medicinal residues add 6 times of distilled water again, soak, decoct, filter with method, twice filtrate is mixed, and heating is concentrated into small size, ultrafiltration membrance filter, adopt 70 ℃ of drying under reduced pressure 36h of vacuum drying oven to get yellow powder, be Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup and fry in shallow oil concentrated 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) Pericarpium Citri Reticulatae water extract preparation
Get the Pericarpium Citri Reticulatae medical material, after the coarse pulverization, sieve, decoct with water 3 times, each 2 hours.The water that adds for the first time 10 times of amounts filters, and medicinal residues add the water of 8 times of amounts for the second time, filters, and adds the water of 6 times of amounts for the third time, filters.3 filtrates are merged, centrifugal (3000rpm) 15 minutes, separation of supernatant, water bath method concentrates, and drying under reduced pressure becomes dry extract again, pulverizes, and crosses 100 mesh sieves, promptly.
(4) Nobiletin preparation method
Get the Pericarpium Citri Reticulatae medical material, coarse pulverization is sieved, and measures the petroleum ether reflux, extract, 5 times for 20 times, each 6h merges petroleum ether extract, behind the recovery petroleum ether, with the mixed solvent heating for dissolving of 2 kinds of solvent compositions of 0.25 times of amount, place, to be crystallized separating out fully, after the filtration, dry under 80 ℃ the Pericarpium Citri Reticulatae total flavones.Measure through the HPLC method, the content of main effective ingredient Nobiletin is 58.4% in the Pericarpium Citri Reticulatae total flavones, and Pericarpium Citri Reticulatae total flavones ethyl acetate extraction concentrates, the utilization polyamide column chromatography, with the mixed solvent eluting of 2 kinds of solvent compositions,, collect and contain the Nobiletin eluent and volatilize solvent in conjunction with thin layer chromatography, recrystallizing methanol, get yellow solid, fusing point 137-138 ℃, be Nobiletin.
(5) choose Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup in proportion and fry in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin, 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 Pericarpium Citri Reticulatae soup and fries in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract.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, illustrates that this natural drug composition toxicity is very little.
5, the inventor fries in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin with Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup originally and makes up, 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 3-5: 2-9: 1-6: produced unexpected synergism during 2-5, potentiation is obvious, extraordinary effect is played in side effect, especially the usage ratio in each component is 5: 9: 6: 5 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 Pericarpium Citri Reticulatae soup is fried in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and each component of Nobiletin and all can be obtained by conventional means, some of them component for example gingerol and Nobiletin 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) Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil the preparation of concentrated extract
According to proportion compatibility is to get Rhizoma Zingiberis Recens, Pericarpium Citri Reticulatae at 1: 1, adds 10 times of amount distilled water, soaks 1h, heating decocts 1h on the electric furnace that adds asbestos gauge, silk filters, and preserves filtrate, and medicinal residues add 6 times of distilled water again, soak, decoct, filter with method, twice filtrate is mixed, and heating is concentrated into small size, ultrafiltration membrance filter, adopt 70 ℃ of drying under reduced pressure 36h of vacuum drying oven to get yellow powder, be Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup and fry in shallow oil concentrated 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) Pericarpium Citri Reticulatae water extract preparation
Get the Pericarpium Citri Reticulatae medical material, after the coarse pulverization, sieve, decoct with water 3 times, each 2 hours.The water that adds for the first time 10 times of amounts filters, and medicinal residues add the water of 8 times of amounts for the second time, filters, and adds the water of 6 times of amounts for the third time, filters.3 filtrates are merged, centrifugal (3000rpm) 15 minutes, separation of supernatant, water bath method concentrates, and drying under reduced pressure becomes dry extract again, pulverizes, and crosses 100 mesh sieves, promptly.
(4) Nobiletin preparation
Get the Pericarpium Citri Reticulatae medical material, coarse pulverization is sieved, and measures the petroleum ether reflux, extract, 5 times for 20 times, each 6h merges petroleum ether extract, behind the recovery petroleum ether, with the mixed solvent heating for dissolving of 2 kinds of solvent compositions of 0.25 times of amount, place, to be crystallized separating out fully, after the filtration, dry under 80 ℃ the Pericarpium Citri Reticulatae total flavones.Measure through the HPLC method, the content of main effective ingredient Nobiletin is 58.4% in the Pericarpium Citri Reticulatae total flavones, and Pericarpium Citri Reticulatae total flavones ethyl acetate extraction concentrates, the utilization polyamide column chromatography, with the mixed solvent eluting of 2 kinds of solvent compositions,, collect and contain the Nobiletin eluent and volatilize solvent in conjunction with thin layer chromatography, recrystallizing methanol, get yellow solid, fusing point 137-138 ℃, be Nobiletin.
Second step: the preparation of preparation
(1) preparation of natural drug composition capsule preparations of the present invention
The Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup that takes by weighing the above-mentioned steps preparation is respectively fried in shallow oil concentrated extract 20mg, gingerol 36mg, Pericarpium Citri Reticulatae water extract 24mg and Nobiletin 20mg, take by weighing starch 345mg, 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
The Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup that takes by weighing the above-mentioned steps preparation is respectively fried in shallow oil concentrated extract 20mg, gingerol 36mg, Pericarpium Citri Reticulatae water extract 24mg and Nobiletin 20mg, take by weighing starch 245mg, 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:
(3) compositions is in following test example: the usage ratio that Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin is 5: 9: 6: 5.The inventor to above-mentioned each composition usage ratio at 3-5: 2-9: 1-6: the compositions that the multiple proportioning combination in the time of between the 2-5 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 it can also be the composition that obtains with conventional means that Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin, 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) Pericarpium Citri Reticulatae is bought and is identified by professor Qian Qiuhai of Shandong Traditional Chinese Medicine University the mature peel of rutaceae orange by the big pharmacy of Qingdao all living creatures.Rhizoma Zingiberis Recens is purchased Yushan Hill degree of conquering east, and is accredited as 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 Pericarpium Citri Reticulatae water extract group, the C group is gingerol and Pericarpium Citri Reticulatae water extract compatibility group (ratio is 3: 2), and the D group is fried in shallow oil concentrated extract for Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup, and (Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin by 5: 9: 6 to the E group: 5 mixed) for natural drug composition of the present invention, 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 Pericarpium Citri Reticulatae water extract 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 Pericarpium Citri Reticulatae water extract 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 Pericarpium Citri Reticulatae soup and fries in shallow oil concentrated 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 reduce the rate of vomitting that causes, alleviates retch and vomit number of times and incubation period is vomitted in effective prolongation.Wherein Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil the concentrated extract group, natural drug composition group of the present invention is vomitted incubation period, compares no significant difference with the ondansetron group.Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil the concentrated extract group, and to cause the rate of vomitting 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.
Table 1 extract compatibility to cisplatin (7.5mg/kg, ip) cause vomit the effect influence (
N=6)
Annotate: a is equivalent to contain the crude drug amount, compares with matched group
bP＜0.05,
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 by being 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 (100,50,25mg/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.And, to S
180, the EAC mice tumor inhibition effect in, the tumor inhibition effect of high dose natural drug composition group is suitable with the positive drug effect.(as table 2,3,4)
Table 2 capsule is to S
180The tumor inhibition effect of mice (
N=12)
Compare with negative control group,
bP＜0.05,
cP＜0.01
Table 3 capsule is to H
22The tumor inhibition effect of mice (
N=12)
Compare with negative control group,
bP＜0.05
Table 4 capsule to the tumor inhibition effect of EAC mice (
N=12)
Compare with negative control group,
bP＜0.05,
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 by being 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.And, to S
180, H
22In the tumor inhibition effect of mice, the tumor inhibition effect of high dose natural drug composition group is remarkable, and is suitable with the positive drug effect.(as table 5,6,7)
Table 5 capsule is to S
180The tumor inhibition effect of mice (
N=12)
Compare with negative control group,
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 (
N=12)
Compare with negative control group,
bP＜0.05
Table 7 capsule to the tumor inhibition effect of EAC mice (
N=12)
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 fries in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin by weight in 3-5 by Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup: the ratio of 2-9: 1-6: 2-5 is formed,
Wherein Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil the preparation method of concentrated extract and is:
According to proportion compatibility is to get Rhizoma Zingiberis Recens, Pericarpium Citri Reticulatae at 1: 1, adds 10 times of amount distilled water, soaks 1h, heating decocts 1h on the electric furnace that adds asbestos gauge, silk filters, and preserves filtrate, and medicinal residues add 6 times of distilled water again, soak, decoct, filter with method, twice filtrate is mixed, and heating is concentrated into small size, ultrafiltration membrance filter, adopt 70 ℃ of drying under reduced pressure 36h of vacuum drying oven to get yellow powder, be Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup and fry in shallow oil concentrated extract.
Wherein the preparation method of Pericarpium Citri Reticulatae water extract is:
Get the Pericarpium Citri Reticulatae medical material, after the coarse pulverization, sieve, decoct with water 3 times, each 2 hours.The water that adds for the first time 10 times of amounts filters, and medicinal residues add the water of 8 times of amounts for the second time, filters, and adds the water of 6 times of amounts for the third time, filters.3 filtrates are merged, centrifugal (3000rpm) 15 minutes, separation of supernatant, water bath method concentrates, and drying under reduced pressure becomes dry extract again, pulverizes, and crosses 100 mesh sieves, promptly.
Wherein the preparation method of Nobiletin is
Get the Pericarpium Citri Reticulatae medical material, coarse pulverization is sieved, and measures the petroleum ether reflux, extract, 5 times for 20 times, each 6h merges petroleum ether extract, behind the recovery petroleum ether, with the mixed solvent heating for dissolving of 2 kinds of solvent compositions of 0.25 times of amount, place, to be crystallized separating out fully, after the filtration, dry under 80 ℃ the Pericarpium Citri Reticulatae total flavones.Measure through the HPLC method, the content of main effective ingredient Nobiletin is 58.4% in the Pericarpium Citri Reticulatae total flavones, and Pericarpium Citri Reticulatae total flavones ethyl acetate extraction concentrates, the utilization polyamide column chromatography, with the mixed solvent eluting of 2 kinds of solvent compositions,, collect and contain the Nobiletin eluent and volatilize solvent in conjunction with thin layer chromatography, recrystallizing methanol, get yellow solid, fusing point 137-138 ℃, be Nobiletin.
2. natural drug composition as claimed in claim 1, wherein to fry in shallow oil the usage ratio of concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin be 5: 9: 6 to Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup: 5.
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, 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.
4. as the preparation method of the natural drug composition of claim 1 or 2, it is characterized in that this method comprises:
(1) Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup is fried in shallow oil the preparation of concentrated extract:
According to proportion compatibility is to get Rhizoma Zingiberis Recens, Pericarpium Citri Reticulatae at 1: 1, adds 10 times of amount distilled water, soaks 1h, heating decocts 1h on the electric furnace that adds asbestos gauge, silk filters, and preserves filtrate, and medicinal residues add 6 times of distilled water again, soak, decoct, filter with method, twice filtrate is mixed, and heating is concentrated into small size, ultrafiltration membrance filter, adopt 70 ℃ of drying under reduced pressure 36h of vacuum drying oven to get yellow powder, be Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup and fry in shallow oil concentrated 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) Pericarpium Citri Reticulatae water extract preparation:
Get the Pericarpium Citri Reticulatae medical material, after the coarse pulverization, sieve, decoct with water 3 times, each 2 hours.The water that adds for the first time 10 times of amounts filters, and medicinal residues add the water of 8 times of amounts for the second time, filters, and adds the water of 6 times of amounts for the third time, filters.3 filtrates are merged, centrifugal (3000rpm) 15 minutes, separation of supernatant, water bath method concentrates, and drying under reduced pressure becomes dry extract again, pulverizes, and crosses 100 mesh sieves, promptly.
(4) the wherein preparation of Nobiletin:
Get the Pericarpium Citri Reticulatae medical material, coarse pulverization is sieved, and measures the petroleum ether reflux, extract, 5 times for 20 times, each 6h merges petroleum ether extract, behind the recovery petroleum ether, with the mixed solvent heating for dissolving of 2 kinds of solvent compositions of 0.25 times of amount, place, to be crystallized separating out fully, after the filtration, dry under 80 ℃ the Pericarpium Citri Reticulatae total flavones.Measure through the HPLC method, the content of main effective ingredient Nobiletin is 58.4% in the Pericarpium Citri Reticulatae total flavones, and Pericarpium Citri Reticulatae total flavones ethyl acetate extraction concentrates, the utilization polyamide column chromatography, with the mixed solvent eluting of 2 kinds of solvent compositions,, collect and contain the Nobiletin eluent and volatilize solvent in conjunction with thin layer chromatography, recrystallizing methanol, get yellow solid, fusing point 137-138 ℃, be Nobiletin.
(5) choose Rhizoma Zingiberis Recens Pericarpium Citri Reticulatae soup in proportion and fry in shallow oil concentrated extract, gingerol, Pericarpium Citri Reticulatae water extract and Nobiletin uniform mixing, 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 played purposes in the potentiation medicine.
7. reduce cancer is put, chemotherapy causes vomiting reaction and the treatment cancer is played purposes in the potentiation medicine in preparation as each described natural drug composition of claim 1-3.
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