CN103845721A - Composition for control of radiation damage or chemotherapy damage and preparation method thereof - Google Patents
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- CN103845721A CN103845721A CN201210498458.5A CN201210498458A CN103845721A CN 103845721 A CN103845721 A CN 103845721A CN 201210498458 A CN201210498458 A CN 201210498458A CN 103845721 A CN103845721 A CN 103845721A
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Abstract
The invention aims to provide a novel composition, which comprises the following raw materials by weight: 1-200 parts of soft-shelled turtle oligopeptide and 1-100 parts of marine fish oligopeptide. The materials further include one or arbitrary combination of 1-100 parts of soybean oligopeptide, 1-100 parts of water soluble dietary fiber, 0.01-0.1 part of zinc gluconate, 0.1-10 parts of a Chinese wolfberry extract, 0.1-10 parts of casein phosphopeptide, 0.01-0.25 part of disodium ethylenediamine tetraacetic acid and 1-500 parts of a taste regulator. The invention also relates to application of the composition to preparation of functional health products, medicines or products for preventing and treating tumor, improving adverse reactions of radioactive therapy and chemotherapy and enhancing immunity.
Description
Technical field
The present invention relates to a kind of peptide combinations, particularly said composition is in the application improving aspect radiation treatment or chemotherapy untoward reaction.
Background technology
At present, the growth of anticancer and the method for transfer mainly contain operation, chemotherapy and radiation, but the toxicity of chemicotherapy is obvious to all, some patients are because toxic reaction and abandoning cure, and some doctors stop the treatment plan to patient because toxic reaction is excessive, and therefore a kind of anticancer of assisting of exploitation is grown and transfer, reduce the injury to health, the medicine or the health food that improve patient's immunity are instant work.
Turtle peptide has higher nutritive value and medical value, as: reduce blood pressure, antioxidant activity, defying age, improves immunity, and antitumor suppresses radiation hazradial bundle etc.
About the purposes of turtle peptide, application number is that 201110236234.2 patent documentations disclose product having anticancer growth and forwarding function and its production and use; Application number is in 201110236297.8 patent documentations, to disclose to have radiation hazradial bundle to be had to product of defencive function and its production and use.
Ocean fish peptide also has higher nutritive value and medical value, as: antioxidant activity, antihypertensive active, defying age, improves immunity, antibacterial activity etc.
About the purposes of ocean fish peptide, application number has significant protective effect for 201110112993.8 patent documentations disclose the skin injury that the compound of marine features oligosaccharide and collagen peptide causes ultraviolet radiation; Application number is the 201110356949.1 patent documentations preparation technology that discloses compound recipe aquatic biological derived collagen peptide calcium composition, utilizes it to prepare the technique of calcium complement agent and the application of calcium complement agent; Application number is a kind of method that 200710014962.2 patent documentations disclose preparing fishskin collagen active peptide by computer auxiliary control directional enzymolysis.
Semen sojae atricolor Toplink promotes mineral absorption, reduces blood pressure, and Semen sojae atricolor Toplink promotes lipid metabolism, and soybean peptide also has immunologic function, and anti-oxidation function reduces blood glucose effect, prevents chemicotherapy damage effect.
About the purposes of soybean peptide, application number is that 200510077612.1 patent documentation discloses and has product of sobering up and liver protecting functions and its production and use, application number is that 200610026655.1 patent documentation discloses a kind of polypeptide and microorganism formulation and preparation method that promotes that cattle tendon is grown, above patent is not all addressed soybean peptide antitumor, prevents and improve the purposes of radiation treatment and chemotherapy untoward reaction.
Li Yingjie etc. " Advance in study of immunomodulation mechanism of Lycium barbarum polysaccharide " (" Chinese Journal of New Drugs ", 2004,13(10)) research show that lycium barbarum polysaccharide can strengthen body immunity; Yang Haijun " functional food ingredient---water soluble dietary fiber " (" Chinese food and nutrition ", 2003, (9)) report water soluble dietary fiber can strengthen gastrointestinal peristalsis; Huang Xiangyuan " factor of phosphopeptide caseinate and promotion mineral absorption thereof " (" food and machinery ", 2008, (3)) report phosphopeptide caseinate can strengthen the absorption of the trace element such as calcium, ferrum.
In application number 200910114806.2 patent documentations, soybean peptide is disclosed, marine fish oligopeptide, zinc gluconate, water soluble dietary fiber, phosphopeptide caseinate compositions are promoting the application of wound healing after operation, but do not address its antitumor, prevent and improve the purposes of radiation treatment and chemotherapy untoward reaction.
In sum, also about molecular weight, turtle peptide, the marine fish peptide combinations of the natural origin below 10000Dalton are not applied to antitumor at present, prevent and improve the report of medicine, health product or the product of radiation treatment and chemotherapy.
Summary of the invention
The object of this invention is to provide a kind of new compositions, said composition has anti-curing oncoma and improves the effect of radiation treatment and chemotherapy untoward reaction, and effect of enhancing immunity; The present composition is at anti-curing oncoma and improve radiation treatment and chemotherapy untoward reaction effect is remarkable.
Compositions of the present invention, raw material comprises: turtle peptide, ocean fish peptide.
The compositions that the present invention is above-mentioned, raw material can further include: one or combination in any in soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator.
Composition material of the present invention is preferred: comprise turtle peptide, ocean fish peptide, soybean peptide.
Composition material of the present invention is preferred: comprise turtle peptide, ocean fish peptide, water soluble dietary fiber.
Compositions preferred feedstock of the present invention: comprise turtle peptide, ocean fish peptide, phosphopeptide caseinate.
Compositions preferred feedstock of the present invention: comprise turtle peptide, ocean fish peptide, soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator.
Compositions of the present invention, preferably includes the raw material of following weight portion: turtle peptide 1-200 part, ocean fish peptide 1-100 part.
Compositions of the present invention, more preferably comprises the raw material of following weight portion: turtle peptide 10-180 part, ocean fish peptide 5-60
Part.
Compositions of the present invention, more preferably comprises the raw material of following weight portion: 150 parts of turtle peptides, 20 parts of ocean fish peptide.
Compositions of the present invention can also add the raw material of following weight parts: one or combination in any in soybean peptide 1-100 part, water soluble dietary fiber 1-100 part, zinc gluconate 0.01-0.1 part, wolfberry fruit extract 0.1-10 part, phosphopeptide caseinate 0.1-10 part, disodiumedetate 0.01-0.25 part, mouthfeel regulator 1-500 part.
The preferably raw material of following weight parts: soybean peptide 10-70 part, water soluble dietary fiber 5-30 part, zinc gluconate
One or combination in any in 0.02-0.08 part, wolfberry fruit extract 0.5-5 part, phosphopeptide caseinate 0.2-2 part, disodiumedetate 0.05-0.22 part, mouthfeel regulator 10-200 part.
More preferably the raw material of following weight parts: one or combination in any in 100 parts of 30 parts of soybean peptide, 15 parts of water soluble dietary fibers, 0.04 part of zinc gluconate, 2 parts of wolfberry fruit extracts, 1 part of phosphopeptide caseinate, 0.1 part of disodiumedetate, mouthfeel regulator.
Compositions of the present invention preferably includes the raw material of following weight portion: turtle peptide 1-200 part, ocean fish peptide 1-100 part,
Soybean peptide 1-100 part.
Preferably include the raw material of following weight portion: turtle peptide 10-180 part, ocean fish peptide 5-60 part, soybean peptide 10-70 part.
More preferably comprise the raw material of following weight portion: 150 parts of turtle peptides, 20 parts of ocean fish peptide, 30 parts of soybean peptide.
Compositions of the present invention preferably includes the raw material of following weight portion: turtle peptide 1-200 part, ocean fish peptide 1-100 part,
Water soluble dietary fiber 1-100 part.
Preferably include the raw material of following weight portion: turtle peptide 10-180 part, ocean fish peptide 5-60 part, water solublity meals fibre
Dimension 5-30 part.
More preferably comprise the raw material of following weight portion: 150 parts of turtle peptides, 20 parts of ocean fish peptide, water soluble dietary fiber 15
Part.
Compositions of the present invention preferably includes the raw material of following weight portion: turtle peptide 1-200 part, ocean fish peptide 1-100 part,
Phosphopeptide caseinate 0.1-10 part.
Preferably include the raw material of following weight portion: turtle peptide 10-180 part, ocean fish peptide 5-60 part, phosphopeptide caseinate
0.2-2 part.
More preferably comprise the raw material of following weight portion: 150 parts of turtle peptides, 20 parts of ocean fish peptide, 1 part of phosphopeptide caseinate.
Compositions of the present invention most preferably comprises the raw material of following weight parts: turtle peptide 1-200 part, ocean fish peptide 1-100
Part, soybean peptide 1-100 part, water soluble dietary fiber 1-100 part, zinc gluconate 0.01-0.1 part, wolfberry fruit extract 0.1-10 part, phosphopeptide caseinate 0.1-10 part, disodiumedetate 0.01-0.25 part, mouthfeel regulator 1-500 part.
More preferably comprise the raw material of following weight parts: turtle peptide 10-180 part, ocean fish peptide 5-60 part, soybean peptide 10-70
Part, water soluble dietary fiber 5-30 part, zinc gluconate 0.02-0.08 part, wolfberry fruit extract 0.5-5 part, phosphopeptide caseinate 0.2-2 part, disodiumedetate 0.05-0.22 part, mouthfeel regulator 10-200 part.
Compositions of the present invention most preferably comprises the raw material of following weight portion: 150 parts of turtle peptides, 20 parts of ocean fish peptide, 30 parts of soybean peptide, 15 parts of water soluble dietary fibers, 0.04 part of zinc gluconate, 2 parts of wolfberry fruit extracts, 1 part of phosphopeptide caseinate, 0.1 part of disodiumedetate, 100 parts of mouthfeel regulators.
Turtle peptide of the present invention is all taking Trionychidae animal Trionyx sinensis Wiegmann Trionyx Sinensis or meat, Carapax Trionycis as primary raw material, can
Being with enzymatic isolation method, acid-hydrolysis method or Production by Microorganism Fermentation, molecular mass are lower than the mixture of the peptide of 10000Dalton; Described enzymatic isolation method carries out enzymolysis including, but not limited to one or more in alkaline protease, trypsin, neutral protease, flavor protease, in present composition and uses thereof, using the object of turtle peptide is to be utilized with peptide mixer form through the Trionyx sinensis Wiegmann of said method processing, retained the nutrition of Trionyx sinensis Wiegmann itself, also make the composition of Trionyx sinensis Wiegmann more easily absorb, therefore enzyme and the processing method thereof that, can process Trionyx sinensis Wiegmann all comprise.Turtle peptide described in the present invention can be prepared by following method: taking Trionyx sinensis Wiegmann as raw material, gill and oils and fats, blend, decoct; Then cooling, add alkaline protease, enzymolysis; By centrifugal enzymolysis solution rear filtration, supernatant is through hollow fiber ultrafiltration film, and rolled film or tubular membrane are refining to be separated.Turtle peptide described in the present invention also can be according to other prior art preparation, for example: Duan Xuchang, Xu Huaide etc. adopt trypsin digestion Trionyx sinensis Wiegmann in " enzymatic liquefaction of Trionyx sinensis Wiegmann and health instant powder research ", disclosed employing pancreatin enzymolysis in the patent document of application number 201010103157.9-boil insulation-complex enzyme zymohydrolysis to process the Trionyx sinensis Wiegmann small-molecular peptides of preparing; Turtle peptide described in the present invention also can directly be buied from the market.
Ocean fish peptide is taking ocean fish skin, fishbone or the flesh of fish as primary raw material, can be with enzymatic isolation method, acid-hydrolysis method or micro-
That biological fermentation process is produced, molecular mass is lower than the mixture of the peptide of 10000Dalton, described enzymatic isolation method is including, but not limited to alkaline protease, trypsin, neutral protease, one or more in flavor protease are carried out enzymolysis, in present composition and uses thereof, using the object of ocean fish peptide is through the ocean of said method processing fish skin, fishbone or the flesh of fish are utilized with peptide mixer form, the fish peptide of ocean described in this description can be prepared according to prior art, for example: Lin Weifeng " Controlled-enzymatic Hydrolysis is prepared the research of biologically active peptide from Sea-fish protein " is by controlling pH value, hydrolysis time, the kind of protease and consumption, the initial concentration of substrate and kind, the molecular weight of hydrolysis temperature and alr mode controlled hydrolysis effect and zymolyte, application number: 200610078017.4 patent document is disclosed prepares collagen peptide with enzymatic isolation method enzyme process from fish skin, fishbone.Soybean peptide soy peptides powder is taking Semen sojae atricolor, bean cake or soybean protein as primary raw material, can be with enzymatic isolation method, acid-hydrolysis method or Production by Microorganism Fermentation, main component is peptide, and the mixture of the peptide of molecular weight distribution below 10000Dalton, described enzymatic isolation method is including, but not limited to alkaline protease, trypsin, neutral protease, one or more in flavor protease are carried out enzymolysis, in present composition and uses thereof, using the object of soybean peptide is the Semen sojae atricolor through said method processing, bean cake or soybean protein are utilized with peptide mixer form, soybean peptide described in this description can be prepared according to prior art, for example: in Hu Aijun " Enzymatic Hydrolysis Technologies of Soybean Protein comparison ", compared the impact of ultrasound wave on enzymolysis degree of hydrolysis, application number: 200680052006.7 patent documents are disclosed prepares soybean peptide mixture with enzymatic isolation method, application number: the disclosed a kind of high-purity of 200510103360.5 patent document, the industrial process of low-molecular-weight soybean oligopeptide powder.Wolfberry fruit extract of the present invention has the effect of enhancing immunity.Add water soluble dietary fiber to strengthen gastrointestinal peristalsis, water soluble dietary fiber described in the present invention includes but not limited to polydextrose, low fat pectin, high fat pectin, apple pectin, pomelo-pectin, blue berry pectin, Fructus Ananadis comosi pectin, oligofructose, oligomeric isomaltose, oligomeric lactose, oligomeric xylose, soybean oligo saccharide, agar powder, carboxymethyl cellulose etc.Add phosphopeptide caseinate casein phosphopeptides, cpp can strengthen the absorption of the trace element such as calcium, ferrum.Wolfberry fruit extract in the present invention is that Fructus Lycii obtains through water extraction and/or alcohol extraction, and Fructus Lycii of the present invention is plant of Solanaceae lycium barbarum
lycium barbarum L.dry mature fruit.
Mouthfeel regulator of the present invention can be one or the combination in any in sweeting agent, acidity regulator, edible essence, and wherein sweeting agent can be one or the combination in any in sucrose, maltose alcohol, sucralose, aspartame, stevioside, cyclamate; Acidity regulator can be one or the combination in any in potassium citrate, citric acid, phosphoric acid, hydrochloric acid, sodium hydroxide; Edible essence can be one or the combination in any in removal agent of raw meat smell essence, polypeptide essence, Mel essence, red cattle essence, flavoring pineapple essence, Folium Bambusae essence, blue berry essence etc.
The invention still further relates to the compositions that contains turtle peptide, ocean fish peptide or the compositions being formed by turtle peptide, the ocean fish peptide purposes in preparation control tumor health product or medicine or product; The invention still further relates to the compositions of the turtle peptide that contains above-mentioned weight portion, ocean fish peptide or the purposes in preparation control tumor health product or medicine or product by the turtle peptide of above-mentioned weight portion, compositions that ocean fish peptide forms.
The invention still further relates to and the compositions that contains turtle peptide, ocean fish peptide or the compositions being formed by turtle peptide, ocean fish peptide improve the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction in preparation; The invention still further relates to the compositions of the turtle peptide that contains above-mentioned weight portion, ocean fish peptide or improve the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction by the turtle peptide of above-mentioned weight portion, compositions that ocean fish peptide forms in preparation.
The invention still further relates to the compositions that contains turtle peptide, ocean fish peptide or the compositions being formed by turtle peptide, ocean fish peptide the purposes in health product or medicine or the product of preparing enhancing immunity; The invention still further relates to the compositions of the turtle peptide that contains above-mentioned weight portion, ocean fish peptide or the purposes in health product or medicine or the product of preparing enhancing immunity by the turtle peptide of above-mentioned weight portion, compositions that ocean fish peptide forms.
The invention still further relates to the compositions being formed by turtle peptide, ocean fish peptide and add soybean peptide, water soluble dietary fiber, Portugal
The purposes of the compositions of a kind of or combination in any composition in grape saccharic acid zinc, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator in preparation control tumor health product or medicine or product; The invention still further relates to by the turtle peptide of above-mentioned weight portion, compositions that ocean fish peptide forms and add a kind of in the soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator of above-mentioned weight portion or compositions that combination in any the forms purposes in preparation control tumor health product or medicine or product.
The invention still further relates to the compositions being formed by turtle peptide, ocean fish peptide and add soybean peptide, water soluble dietary fiber, Portugal
The compositions of a kind of or combination in any in grape saccharic acid zinc, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator composition is improved the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction in preparation; The invention still further relates to by the turtle peptide of above-mentioned weight portion, compositions that ocean fish peptide forms adds a kind of in the soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator of above-mentioned weight portion or compositions that combination in any forms to improve the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction in preparation.
The invention still further relates to the compositions being formed by turtle peptide, ocean fish peptide and add soybean peptide, water soluble dietary fiber, Portugal
The purposes of the compositions of a kind of or combination in any composition in grape saccharic acid zinc, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator in health product or medicine or the product of preparing enhancing immunity; The invention still further relates to by the turtle peptide of above-mentioned weight portion, compositions that ocean fish peptide forms and add a kind of in the soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator of above-mentioned weight portion or compositions that combination in any the forms purposes in health product or medicine or the product of preparing enhancing immunity.
The invention still further relates to the compositions that contains turtle peptide, ocean fish peptide, soybean peptide or the compositions being formed by turtle peptide, ocean fish peptide, the soybean peptide purposes in preparation control tumor health product or medicine or product; The invention still further relates to the turtle peptide, ocean fish peptide, the compositions of soybean peptide or the compositions being formed by turtle peptide, ocean fish peptide, the soybean peptide of above-mentioned weight portion that contain above-mentioned weight portion and prevent and treat the purposes in tumor health product or medicine or product in preparation.
The invention still further relates to and the compositions that contains turtle peptide, ocean fish peptide, soybean peptide or the compositions being formed by turtle peptide, ocean fish peptide, soybean peptide improve the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction in preparation; The compositions that the invention still further relates to the compositions of the turtle peptide that contains above-mentioned weight portion, ocean fish peptide, soybean peptide or be made up of turtle peptide, ocean fish peptide, the soybean peptide of above-mentioned weight portion is improved the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction in preparation.
The invention still further relates to the compositions that contains turtle peptide, ocean fish peptide, soybean peptide or the compositions being formed by turtle peptide, ocean fish peptide, soybean peptide the purposes in health product or medicine or the product of preparing enhancing immunity; The purposes of the compositions that the invention still further relates to the compositions of the turtle peptide that contains above-mentioned weight portion, ocean fish peptide, soybean peptide or formed by turtle peptide, ocean fish peptide, the soybean peptide of above-mentioned weight portion in health product or medicine or the product of preparing enhancing immunity.
The invention still further relates to the compositions that contains turtle peptide, ocean fish peptide, water soluble dietary fiber or the compositions being formed by turtle peptide, ocean fish peptide, the water soluble dietary fiber purposes in preparation control tumor health product or medicine or product; The invention still further relates to the turtle peptide, ocean fish peptide, the compositions of water soluble dietary fiber or the compositions being formed by turtle peptide, ocean fish peptide, the water soluble dietary fiber of above-mentioned weight portion that contain above-mentioned weight portion and prevent and treat the purposes in tumor health product or medicine or product in preparation.
The invention still further relates to and the compositions that contains turtle peptide, ocean fish peptide, water soluble dietary fiber or the compositions being formed by turtle peptide, ocean fish peptide, water soluble dietary fiber improve the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction in preparation; The compositions that the invention still further relates to the compositions of the turtle peptide that contains above-mentioned weight portion, ocean fish peptide, water soluble dietary fiber or be made up of turtle peptide, ocean fish peptide, the water soluble dietary fiber of above-mentioned weight portion is improved the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction in preparation.
The invention still further relates to the compositions that contains turtle peptide, ocean fish peptide, water soluble dietary fiber or the compositions being formed by turtle peptide, ocean fish peptide, water soluble dietary fiber the purposes in health product or medicine or the product of preparing enhancing immunity; The purposes of the compositions that the invention still further relates to the compositions of the turtle peptide that contains above-mentioned weight portion, ocean fish peptide, water soluble dietary fiber or formed by turtle peptide, ocean fish peptide, the water soluble dietary fiber of above-mentioned weight portion in health product or medicine or the product of preparing enhancing immunity.
The invention still further relates to the compositions that contains turtle peptide, ocean fish peptide, phosphopeptide caseinate or the compositions being formed by turtle peptide, ocean fish peptide, the phosphopeptide caseinate purposes in preparation control tumor health product or medicine or product; The invention still further relates to the turtle peptide, ocean fish peptide, the compositions of phosphopeptide caseinate or the compositions being formed by turtle peptide, ocean fish peptide, the phosphopeptide caseinate of above-mentioned weight portion that contain above-mentioned weight portion and prevent and treat the purposes in tumor health product or medicine or product in preparation.
The invention still further relates to and the compositions that contains turtle peptide, ocean fish peptide, phosphopeptide caseinate or the compositions being formed by turtle peptide, ocean fish peptide, phosphopeptide caseinate improve the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction in preparation; The invention still further relates to the turtle peptide, ocean fish peptide, the casein phosphoric acid that contain above-mentioned weight portion
The compositions of peptide or the compositions being made up of turtle peptide, ocean fish peptide, the phosphopeptide caseinate of above-mentioned weight portion change in preparation
Purposes in health product or medicine or the product of kind radiation treatment and chemotherapy untoward reaction.
The invention still further relates to the compositions that contains turtle peptide, ocean fish peptide, phosphopeptide caseinate or the compositions being formed by turtle peptide, ocean fish peptide, phosphopeptide caseinate the purposes in health product or medicine or the product of preparing enhancing immunity; The purposes of the compositions that the invention still further relates to the compositions of the turtle peptide that contains above-mentioned weight portion, ocean fish peptide, phosphopeptide caseinate or formed by turtle peptide, ocean fish peptide, the phosphopeptide caseinate of above-mentioned weight portion in health product or medicine or the product of preparing enhancing immunity.
The invention still further relates to by turtle peptide, ocean fish peptide, soybean peptide, water soluble dietary fiber, zinc gluconate, Fructus Lycii
The purposes of the compositions of extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator composition in preparation control tumor health product or medicine or product; The invention still further relates to the purposes of the compositions being formed by turtle peptide, ocean fish peptide, soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, the mouthfeel regulator of above-mentioned weight portion in preparation control tumor health product or medicine or product.
The invention still further relates to the compositions that formed by turtle peptide, ocean fish peptide, soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator and improve the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction in preparation; The invention still further relates to the compositions that formed by turtle peptide, ocean fish peptide, soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, the mouthfeel regulator of above-mentioned weight portion and improve the purposes in health product or medicine or the product of radiation treatment and chemotherapy untoward reaction in preparation.
The invention still further relates to by turtle peptide, ocean fish peptide, soybean peptide, water soluble dietary fiber, zinc gluconate, Fructus Lycii
The purposes of the compositions of extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator composition in health product or medicine or the product of preparing enhancing immunity; The invention still further relates to the purposes of the compositions being formed by turtle peptide, ocean fish peptide, soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, the mouthfeel regulator of above-mentioned weight portion in health product or medicine or the product of preparing enhancing immunity.
The untoward reaction of chemicotherapy of the present invention mainly contains: (1) hemopoietic system symptom (bone marrow depression): leukopenia, anemia, thrombocytopenia; (2) symptom of digestive tract: feel sick, vomiting, anorexia, constipation, diarrhoea; (3) mucosal injury: stomatitis, oral ulcer, esophagitis; (4) liver, the heart, lung, nephrotoxicity; (5) neurotoxicity: nervus centralis obstacle, nerve ending; (6) skin lesion; (7) alopecia; (8) other: gonad infringement, second primary cancer etc.
Compositions of the present invention can be made into any one in oral liquid, granule, capsule, tablet, powder, dispersible tablet, syrup.
The present composition follows these steps to be prepared into oral liquid:
1. preparation: take supplementary material by prescription, standardize solution adds water;
2. filter: with 30 μ m-0.1 μ m filter element filterings, fill;
3. sterilizing: 100-125 DEG C, sterilizing 5-60 minute, to obtain final product.
In above-mentioned steps 2, also can select 10 μ m-0.15 μ m filter element filtering, preferably 0.2 μ m filter element filterings; In step 3, preferably 102-121 DEG C, sterilizing 8-50 minute; More preferably 105 DEG C, sterilizing 30 minutes;
It is low that this product has cost, good effect, the natural origin feature that has no side effect.
In order to understand better the present invention, below by the preparation with the embodiment formal description present composition.The turtle peptide, soybean peptide, the marine fish peptide molecular weight that in embodiment, use are less than 10000Dalton.The zoopery of the compositions being made up of turtle peptide, ocean fish peptide by embodiment 64 and result illustrate that it has anti-curing oncoma, improves radiation treatment and chemotherapy untoward reaction, the effect of enhancing immunity.Zoopery by the compositions being made up of turtle peptide, soybean peptide, ocean fish peptide with embodiment 61 and result illustrate that it has anti-curing oncoma, improves radiation treatment and chemotherapy untoward reaction, the effect of enhancing immunity.These embodiment are just in order to describe object of the present invention, instead of in office where face limits the scope of the invention.
The compositions being similarly made up of turtle peptide, ocean fish peptide adds soybean peptide, water soluble dietary fiber, gluconic acid
The compositions of a kind of or combination in any composition in zinc, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator all can reach and have same pharmacological action.
Detailed description of the invention
Embodiment 1
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, water soluble dietary fiber 15kg, wolfberry fruit extract 2kg, disodiumedetate 0.03kg, citric acid 2.5kg, potassium citrate 1.5kg, maltose alcohol 90kg, sucralose 0.25kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 1.4kg, in preparing tank, add 500L purified water, add successively disodiumedetate, turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltose alcohol, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, by sucralose, phosphopeptide caseinate, after zinc gluconate dissolves by appropriate purified water, join in filtrate, add water to 1000L, stir 15 minutes, add essence, stir again 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 2
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, after stirring is dissolved it, be that 5 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.1 μ m filter filtration, fill, every bottle of 100ml, 121 DEG C of sterilizings 8 minutes, to obtain final product.
Embodiment 3
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, after stirring is dissolved it, be that 30 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 5 μ m filters filtrations, fill, every bottle of 100ml, 105 DEG C of sterilizings 60 minutes, to obtain final product.
Embodiment 4
Turtle peptide 10kg, soybean peptide 10kg, ocean fish peptide 5kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 5
Turtle peptide 180kg, soybean peptide 70kg, ocean fish peptide 60kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 6
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 7
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, water soluble dietary fiber 1kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, after stirring is dissolved it, be that 5 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.15 μ m filter filtration, fill, every bottle of 100ml, 102 DEG C of sterilizings 50 minutes, to obtain final product.
Embodiment 8
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg, water soluble dietary fiber 150kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 9
Turtle peptide 10kg, soybean peptide 10kg, ocean fish peptide 5kg, water soluble dietary fiber (being purchased from promise Shen, Shanghai food commerce and trade company) 5kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 10
Turtle peptide 180kg, soybean peptide 70kg, ocean fish peptide 60kg, water soluble dietary fiber 30kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 11
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, water soluble dietary fiber 15kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 12
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, zinc gluconate 0.01kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, zinc gluconate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 13
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg, zinc gluconate 0.1kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, zinc gluconate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 14
Turtle peptide 10kg, soybean peptide 10kg, ocean fish peptide 5kg, zinc gluconate 0.02kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, zinc gluconate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 15
Turtle peptide 180kg, soybean peptide 70kg, ocean fish peptide 60kg, zinc gluconate 0.08kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, zinc gluconate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 16
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, zinc gluconate 0.05kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, zinc gluconate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 17
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, wolfberry fruit extract 0.1kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, wolfberry fruit extract, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 18
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg, wolfberry fruit extract 20kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, wolfberry fruit extract, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 19
Turtle peptide 10kg, soybean peptide 10kg, ocean fish peptide 5kg, wolfberry fruit extract 0.5kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, wolfberry fruit extract, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 20
Turtle peptide 180kg, soybean peptide 70kg, ocean fish peptide 60kg, wolfberry fruit extract 5kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, wolfberry fruit extract, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 21
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, wolfberry fruit extract 2kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, wolfberry fruit extract, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 22
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, phosphopeptide caseinate 0.1kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 23
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg, phosphopeptide caseinate 10kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 24
Turtle peptide 10kg, soybean peptide 10kg, ocean fish peptide 5kg, phosphopeptide caseinate 0.2kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 25
Turtle peptide 180kg, soybean peptide 70kg, ocean fish peptide 60kg, phosphopeptide caseinate 2kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 26
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, phosphopeptide caseinate 0.25kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 27
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, maltose alcohol 1kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, maltose alcohol, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 28
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg, maltose alcohol 350kg, citric acid 80kg, disodiumedetate 0.25kg, potassium citrate 67kg, sucralose 0.25kg, essence 2.5kg adds 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, maltose alcohol, citric acid, disodiumedetate, potassium citrate, sucralose, after essence stirs it is dissolved, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 29
Turtle peptide 10kg, soybean peptide 10kg, ocean fish peptide 5kg, maltose alcohol 10kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, maltose alcohol, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 30
Turtle peptide 180kg, soybean peptide 70kg, ocean fish peptide 60kg, maltose alcohol 150kg, citric acid 28kg, disodiumedetate 0.25kg, potassium citrate 20kg, sucralose 0.25kg, essence 1.5kg adds 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, maltose alcohol, citric acid, disodiumedetate, potassium citrate, sucralose, after essence stirs it is dissolved, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 31
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, maltose alcohol 90kg, citric acid 5kg, disodiumedetate 0.25kg, potassium citrate 3kg, sucralose 0.25kg, essence 1.5kg adds 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, maltose alcohol, citric acid, disodiumedetate, potassium citrate, sucralose, after essence stirs it is dissolved, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 32
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, water soluble dietary fiber 1kg, wolfberry fruit extract 0.1kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, after stirring is dissolved it, be that 30 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.1 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 33
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg, water soluble dietary fiber 150kg, wolfberry fruit extract 20kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, stirs and makes after its dissolving, is that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 34
Turtle peptide 10kg, soybean peptide 10kg, ocean fish peptide 5kg, water soluble dietary fiber 5kg, wolfberry fruit extract 0.5kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, stirs and makes after its dissolving, is that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 35
Turtle peptide 180kg, soybean peptide 70kg, ocean fish peptide 60kg, water soluble dietary fiber 30kg, wolfberry fruit extract 5kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 36
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, water soluble dietary fiber 15kg, wolfberry fruit extract 20kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 37
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, wolfberry fruit extract 0.1kg, phosphopeptide caseinate 0.1kg add 500L purified water in preparing tank, adding successively turtle peptide, soybean peptide, ocean fish peptide, wolfberry fruit extract, phosphopeptide caseinate, stir and make after its dissolving, is that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 38
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg, wolfberry fruit extract 20kg, phosphopeptide caseinate 10kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, wolfberry fruit extract, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 39
Turtle peptide 10kg, soybean peptide 10kg, ocean fish peptide 5kg, wolfberry fruit extract 0.5kg, phosphopeptide caseinate 0.2kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, wolfberry fruit extract, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 40
Turtle peptide 180kg, soybean peptide 70kg, ocean fish peptide 60kg, wolfberry fruit extract 5kg, phosphopeptide caseinate 2kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, wolfberry fruit extract, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 41
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, wolfberry fruit extract 2kg, phosphopeptide caseinate 0.25kg add 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, wolfberry fruit extract, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, then stir 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, to obtain final product.
Embodiment 42
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, water soluble dietary fiber 1kg, wolfberry fruit extract 0.1kg, zinc gluconate 0.01kg, phosphopeptide caseinate 0.1kg, maltose alcohol 1kg, in preparing tank, add 500L purified water, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, zinc gluconate, phosphopeptide caseinate, maltose alcohol, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 121 DEG C of sterilizings 8 minutes, obtain.
Embodiment 43
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg, water soluble dietary fiber 150kg, wolfberry fruit extract 20kg, zinc gluconate 0.1kg, phosphopeptide caseinate 10kg, maltose alcohol 350kg, citric acid 5Kg, disodiumedetate 0.01kg, potassium citrate 3kg, sucralose 0.25kg, essence 1.5kg adds 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, zinc gluconate, phosphopeptide caseinate, maltose alcohol, citric acid, disodiumedetate, potassium citrate, sucralose, after essence stirs it is dissolved, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 100 DEG C of sterilizings 60 minutes, obtain.
Embodiment 44
Turtle peptide 10kg, soybean peptide 10kg, ocean fish peptide 5kg, water soluble dietary fiber 5kg, wolfberry fruit extract 0.5kg, zinc gluconate 0.02kg, phosphopeptide caseinate 0.2kg, maltose alcohol 10kg, in preparing tank, add 500L purified water, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, zinc gluconate, phosphopeptide caseinate, after maltose alcohol 1kg stirs it is dissolved, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 45
Turtle peptide 180kg, soybean peptide 70kg, ocean fish peptide 60kg, water soluble dietary fiber 30kg, wolfberry fruit extract 5kg, zinc gluconate 0.08kg, phosphopeptide caseinate 2kg, maltose alcohol 150kg, citric acid 28kg, disodiumedetate 0.025kg, potassium citrate 20kg, sucralose 0.25kg, essence 1.5kg adds 500L purified water in preparing tank, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, zinc gluconate, phosphopeptide caseinate, maltose alcohol, citric acid, disodiumedetate, potassium citrate, sucralose, after essence stirs it is dissolved, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 46
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, water soluble dietary fiber 15kg, wolfberry fruit extract 2kg, citric acid 2.5kg, disodiumedetate 0.22kg, potassium citrate 1.5kg, maltose alcohol 90kg, sucralose 0.25kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 1.4kg, in preparing tank, add 500L purified water, add successively disodiumedetate, turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltose alcohol, after stirring is dissolved it, be that 0.45 μ m filter filters with aperture, obtain filtrate, by sucralose, phosphopeptide caseinate, after zinc gluconate dissolves by appropriate purified water, join in filtrate, add water to 1000L, stir 15 minutes, add essence, stir again 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 47
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, water soluble dietary fiber 1kg, wolfberry fruit extract 0.1kg, zinc gluconate 0.01kg, phosphopeptide caseinate 0.1kg, in preparing tank, add 500L purified water, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, zinc gluconate, phosphopeptide caseinate, after stirring is dissolved it, be that 0.45 μ m filter filters with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.1 μ m filter filtration, fill, every bottle of 100ml, 102 DEG C of sterilizings 60 minutes, obtain.
Embodiment 48
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg, water soluble dietary fiber 150kg, wolfberry fruit extract 20kg, zinc gluconate 0.1kg, phosphopeptide caseinate 10kg, in preparing tank, add 500L purified water, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, zinc gluconate, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 49
Turtle peptide 10kg, soybean peptide 10kg, ocean fish peptide 5kg, water soluble dietary fiber 5kg, wolfberry fruit extract 0.5kg, zinc gluconate 0.02kg, phosphopeptide caseinate 0.2kg, in preparing tank, add 500L purified water, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, zinc gluconate, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 50
Turtle peptide 180kg, soybean peptide 70kg, ocean fish peptide 60kg, water soluble dietary fiber 30kg, wolfberry fruit extract 5kg, zinc gluconate 0.08kg, phosphopeptide caseinate 5kg, in preparing tank, add 500L purified water, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, zinc gluconate, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 51
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, water soluble dietary fiber 15kg, wolfberry fruit extract 2kg, zinc gluconate 0.04kg, phosphopeptide caseinate 0.25kg, in preparing tank, add 500L purified water, add successively turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, zinc gluconate, phosphopeptide caseinate, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, add water to 1000L, stir 15 minutes, stir again 10 minutes, with 0.2 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 52
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, water soluble dietary fiber 15kg, wolfberry fruit extract 2kg, citric acid 2.5kg, disodiumedetate 0.03kg, potassium citrate 1.5kg, maltose alcohol 90kg, sucralose 0.20kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 1.4kg, in preparing tank, add 500L purified water, add successively disodiumedetate, turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltose alcohol, after stirring is dissolved it, be that 10 μ m filters filter with aperture, obtain filtrate, by sucralose, phosphopeptide caseinate, after zinc gluconate dissolves by appropriate purified water, join in filtrate, add water to 1000L, stir 15 minutes, add essence, stir again 10 minutes, with 0.45 μ m filter filtration, fill, every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
Embodiment 53
Turtle peptide 200kg, soybean peptide 100kg, ocean fish peptide 200kg.Water soluble dietary fiber 150kg, wolfberry fruit extract 20kg, citric acid 3.0kg, potassium citrate 2.0kg, maltodextrin 120kg, phosphopeptide caseinate 10kg, zinc gluconate 0.1 kg, essence 2kg, in blender, add anti-turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltodextrin, phosphopeptide caseinate, zinc gluconate carries out batch mixing, the material mixing is carried out to boiling granulating with 10% aqueous solution of PVP K30 as binding agent, control atomizing pressure is 0.15MPa, temperature of charge is 45 DEG C, inlet temperature is 75 DEG C, 20 mesh sieve granulate for gained granule, add essence, use aluminum aluminum to pack, 10 grams every bag.
Embodiment 54
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, water soluble dietary fiber 15kg, wolfberry fruit extract 2kg, citric acid 2.5kg, potassium citrate 1.5kg, maltodextrin 90kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 1.2kg, in blender, add turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltodextrin, phosphopeptide caseinate, zinc gluconate carries out batch mixing, the material mixing is carried out to boiling granulating with 10% aqueous solution of PVP K30 as binding agent, control atomizing pressure is 0.15MPa, temperature of charge is 45 DEG C, inlet temperature is 75 DEG C, 30 mesh sieve granulate for gained granule, add essence, pack with aluminum aluminum, 10 grams every bag, obtain powder.
Embodiment 55
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, water soluble dietary fiber 1kg, wolfberry fruit extract 2kg, citric acid 1.5kg, potassium citrate 1kg, maltodextrin 75kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 1.0kg, in blender, add turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltodextrin, phosphopeptide caseinate, zinc gluconate carries out batch mixing, the material mixing is carried out to boiling granulating with 10% aqueous solution of PVP K30 as binding agent, control atomizing pressure is 0.15MPa, temperature of charge is 45 DEG C, inlet temperature is 75 DEG C, 30 mesh sieve granulate for gained granule, add essence, pack with aluminum aluminum, restrain to obtain granule for every bag 10.
Embodiment 56
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, water soluble dietary fiber 1kg, wolfberry fruit extract 2kg, citric acid 1.5kg, potassium citrate 1kg, maltodextrin 75kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 0.5kg, in blender, add turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltodextrin, phosphopeptide caseinate, zinc gluconate carries out batch mixing, the material mixing is carried out to boiling granulating with 10% aqueous solution of PVP K30 as binding agent, control atomizing pressure is 0.15MPa, temperature of charge is 45 DEG C, inlet temperature is 75 DEG C, 30 mesh sieve granulate for gained granule, add essence, encapsulating capsule.
Embodiment 57
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, water soluble dietary fiber 1kg, wolfberry fruit extract 2kg, citric acid 1.5kg, potassium citrate 1kg, maltodextrin 1.5kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 0.5kg, in blender, add turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltodextrin, phosphopeptide caseinate, zinc gluconate to carry out batch mixing, the material mixing is granulated as binding agent with 10% aqueous solution of PVP K30; 30 mesh sieve granulate for gained granule, add essence, magnesium stearate, tabletting.
Embodiment 58
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, water soluble dietary fiber 1kg, wolfberry fruit extract 2kg, citric acid 1.5kg, potassium citrate 1kg, maltodextrin 1.5kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 0.5kg, in blender, add turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltodextrin, phosphopeptide caseinate, zinc gluconate to carry out batch mixing, the material mixing is granulated as binding agent with 10% aqueous solution of PVP K30; 30 mesh sieve granulate for gained granule, add essence, MCC, PVPP
xL, tabletting, obtains dispersible tablet.
Embodiment 59
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, water soluble dietary fiber 1kg, wolfberry fruit extract 2kg, citric acid 1.5kg, potassium citrate 1kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 0.5kg, in blender, add turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltodextrin, phosphopeptide caseinate, zinc gluconate to carry out batch mixing, by the material maltodextrin mixing, add appropriate binding agent or wetting agent, make pill.
Embodiment 60
Turtle peptide 1kg, soybean peptide 1kg, ocean fish peptide 1kg, water soluble dietary fiber 1kg, wolfberry fruit extract 2kg, citric acid 1.5kg, potassium citrate 1kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 0.5kg, in blender, add turtle peptide, soybean peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltodextrin, phosphopeptide caseinate, zinc gluconate to carry out batch mixing, by the material mixing, add appropriate sucrose and antiseptic, make syrup.
Embodiment 61
Experiment material:
Turtle peptide: Jiangzhong Pharmaceutical Co., Ltd. provides
Soybean peptide: Hai Shi bio tech ltd, Zhejiang product batch number: 20110312
Ocean fish peptide: Hai Shi bio tech ltd, Zhejiang product batch number: 20120318
Take turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg, in blender, add turtle peptide, soybean peptide, ocean fish peptide to carry out batch mixing, use aluminum aluminum to pack, 10 grams every bag.
Carry out following Ergonomy experiment with the sample of embodiment 61 sample preparations:
One, antineoplastic efficacy study
1. experiment material
1.1 animals: BALB/c mice, 18～22g, clean level, purchased from Hunan Si Laike Jing Da laboratory animal company limited, the quality certification number: SCXK(Hunan) 2009-0004.The strain of murine sarcoma S180 tumor, Lewis pulmonary carcinoma, BGC-823 gastric carcinoma cells cell strain, purchased from Chinese Academy of Medical Sciences's cell bank, gone down to posterity, taken care of by Jiangxi College of Traditional Chinese Medicine pharmacology subject group.
1.2 medicines and reagent: embodiment, 61 samples: provided by Jiangzhong Pharmaceutical Co., Ltd., be mixed with every milliliter of medicinal liquid 0.286g crude drug that is equivalent to embodiment 61 samples with front with sterile purified water; 5-fluorouracil injection (5-FU, Shandong Pharmaceutical all together), concanavalin A, Con A (ConA, Sigma company), MTT(Sigma company), IL-2, TNF-α ELISA detection kit be all purchased from ExCell company.
1.3 key instrument NanoVue trace albumin nucleic acid instrument (GE company); AL204 electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd.); XT-2000i automated blood analysis (Japanese Sysmex company); ELx800 type microplate reader (BioTek, the U.S.).
2. method
2.1 dosage designs: everyone recommended intake every day of embodiment 61 samples is: 20g crude drug/70kg body weight, extrapolate thus mice daily intake and be: low dose group, 1.43g crude drug/kg; Middle dosage group, 2.86g crude drug/kg; High dose group, 5.72g crude drug/kg, is equivalent to respectively 5,10 and 20 times of people's daily intake.
The tumor-inhibiting action of 2.2 embodiment 61 samples to murine sarcoma S180: the mice cervical vertebra dislocation of 8 days after intraperitoneal inoculation murine sarcoma S180 cell is put to death, take out ascites under aseptic condition, with normal saline 1:2 dilution.Mice is divided into 4 groups at random by body weight, 10 every group, sets up model control group, the basic, normal, high dosage group of embodiment 61 sample.Give every mouse armpit subcutaneous vaccination tumor liquid 0.2mL, next day after inoculation, the basic, normal, high dosage of embodiment 61 sample is given the embodiment 61 sample medicinal liquids of pressing respectively 1.43g crude drug/kg body weight, 2.86g crude drug/kg body weight, the basic, normal, high concentration of 5.72g crude drug/kg body weight gavage, every day 1 time, successive administration 30 days.After last administration 24 hours, animal was put to death in cervical vertebra dislocation, and point another name tumor weight, calculates inhibition rate of tumor growth (%).Computing formula is: the flat tumor weight of average tumor weight-treatment group is all organized in inhibition rate of tumor growth %=(contrast) the average tumor of ÷ matched group heavy × 100%.
The tumor-inhibiting action of 2.3 embodiment 61 samples to mice transplanted tumor Lewis pulmonary carcinoma: with the mice transplanted tumor well-grown tumor of Lewis pulmonary carcinoma source, under aseptic condition, get the solid tumor tissue of tumor animal, be cut into fine grained chippings with shears, transplant or add physiological saline solution with glass homogenizer with the trocar and wear into homogenate, count the oncocyte number in every mL homogenate, give every animal subcutaneous vaccination 2 × 10
6cell.Grouping administration is with 2.3, and after last administration 24 hours, animal was put to death in cervical vertebra dislocation, and point another name tumor weight, calculates inhibition rate of tumor growth (%).
2.4 impacts of embodiment 61 samples on tumor-bearing mice immunologic function
2.4.1 the impact of embodiment 61 samples on tumor-bearing mice lymhocyte transformation rate: inoculation tumor liquid and administration are with 2.2 and 2.3.Under aseptic condition, get mouse spleen, make single cell suspension, after splitting erythrocyte, adjust splenocyte density with the RPMI-1640 culture fluid containing 10% calf serum, make 5 × 10
6/ ml mouse boosting cell suspension, adds every a splenocyte suspension respectively in two 96 orifice plates, and wherein a hole adds 75 μ l ConA, and another hole adds equivalent culture medium in contrast, and above-mentioned each sample repeats 3 multiple holes, and 96 orifice plates are placed at 37 DEG C 5% CO
2in incubator, cultivate 72h; Cultivation finishes front 4h, and every hole adds MTT (5mg/ml) 50 μ l, continues to cultivate, after cultivation finishes, abandon supernatant, every hole adds 150 μ l acid isopropyl alcohol, mix, measure the optical density at 570nm wavelength place by microplate reader, calculate splenocyte turn over number.
Splenocyte turn over number=ConA stimulation hole OD value-stimulate hole OD value without ConA
2.4.2 the affect double antibody sandwich ELISA of embodiment 61 samples on S180 mice serum TNF-α and IL-2 content, to detect TNF-α as example: with anti-mice TNF-α mAb coated elisa plate, TNF-α and mAb in standard substance and sample are mixed to form to immune complex, add successively the affinity element of biotinylated anti-mice TNF-Alpha antibodies and horseradish peroxidase-labeled, finally add zymolyte OPD colour developing, measure OD value at wavelength 450nm place by microplate reader, and drawing standard curve, the detecting step of IL-2 is substantially the same.
The impact of the potentiation of 2.5 embodiment 61 samples on the chemotherapy of people's Transplanted Gastric Carcinoma mice: mice is in back intradermal vaccination BGC-823 gastric carcinoma cells 1 × 10
7/ only, when gross tumor volume grows to 1 cm
3, under aseptic condition, take off part tumor piece when the left and right, be cut into 1 mm
3the tumor piece of left and right, is inoculated into mouse back with the trocar subcutaneous, forms subcutaneous transplantation tumor, is divided at random five groups: 1. blank group, only gives equal-volume solvent after inoculation 48 h; 2. 5-FU treatment group, from after inoculation the 28th day, lumbar injection 5-FU, dosage is 25 mg/kg, every day 1 time, continuous 14 days.3. 5-FU+ low dosage embodiment 61 sample sets started to give 5-FU from after inoculation the 28th day, the same 2. group of dosage, and embodiment 61 sample 1.43g/kg every day, after inoculation the 7th day starts administration, continuous 35 days, every day 1 time; 4. dose delivery example 61 sample sets in 5-FU+ started to give 5-FU from after inoculation the 28th day, the same 2. group of dosage, and embodiment 61 sample 2.86g/kg every day, after inoculation the 7th day starts administration, continuous 35 days, every day 1 time; 5. 5-FU+ high dose embodiment 61 sample sets started to give 5-FU from after inoculation the 28th day, the same 2. group of dosage, and embodiment 61 sample 5.72g/kg every day, after inoculation the 7th day starts administration, continuous 35 days, every day 1 time.Above-mentioned each treated animal finishes experiment after 5 weeks, peel off tumor tissues and weigh and calculate tumor control rate.
The impact of the Attenuation of 2.6 embodiment 61 samples on the chemotherapy of people's Transplanted Gastric Carcinoma mice: grouping and administration same 2.5.Above-mentioned lotus tumor is in the blood sampling of last administration 24h posterior orbit venous plexus, and Automatic Blood Cell Analyzer detects mouse peripheral blood cell, measures numeration of leukocyte in blood, and mice is put to death in cervical vertebra dislocation afterwards, peels off right side femur, detects mouse bone marrow cells DNA content.
2.7 statistical method: experimental data represents with `X ± S, adopts one factor analysis of variance, and P<0.05 is judged as difference and has significance.
3 results
3.1 embodiment 61 samples are to mice S180 and Lewis pulmonary carcinoma tumor-inhibiting action
The results are shown in Table 1.With model control group comparison, the middle and high dosage of embodiment 61 sample has stronger inhibitory action to murine sarcoma S180, and suppression ratio is respectively 24.0% and 50.0%.The middle and high dosage of embodiment 61 sample has stronger inhibitory action to Mice Bearing Lewis pulmonary carcinoma, and suppression ratio is respectively 34.85% and 43.57%.
3.2 embodiment 61 samples on tumor-bearing mice immunologic function impact
3.2.1 the impact of embodiment 61 samples on transplanted tumor S180 mice, Lewis lung cancer in mice splenocyte conversion ratio
The results are shown in Table 2.Compared with model control group, the middle and high dosage of embodiment 61 sample all has stronger raising effect to mice transplanted tumor S180 mice, Lewis lung cancer in mice splenocyte conversion ratio.
3.2.2 the impact of embodiment 61 samples on transplanted tumor S180 mice serum TNF-α and IL-2 content
The results are shown in Table 3.With model control group comparison, the middle and high dosage of embodiment 61 sample all has stronger raising effect to transplanted tumor S180 mice serum TNF-α and IL-2 content.
The potentiation of 3.3 embodiment 61 samples to people's Transplanted Gastric Carcinoma mice 5-FU chemotherapy
The results are shown in Table 4.After embodiment 61 samples and 5-FU therapeutic alliance, the growth of tumor is subject to obvious inhibition, there is statistical significance compared with model control group, compared with the independent treatment group of 5-FU, the tumor weight average of each therapeutic alliance group has reduction in various degree, tumor growth is subject to further inhibition, and presents dose dependent, but does not occur statistical significance.
The impact of 3.4 embodiment 61 samples on people's Transplanted Gastric Carcinoma mice 5-FU chemotherapy Attenuation
The results are shown in Table 5.Tumor animal is after 5-FU successive administration 14 days; there is obvious bone marrow inhibition in animal subject; leukocyte counts and bone marrow DNA content are all significantly lower than matched group; and animal subject is giving after 5 weeks embodiment 61 samples continuously; leukocyte counts and bone marrow DNA content all have rebound significantly, and embodiment 61 samples have obvious bone marrow protective effect.
4. conclusion:
Animal experiment study shows: the middle and high dosage of embodiment 61 sample has stronger inhibitory action to murine sarcoma S180 and mice transplanted tumor Lewis pulmonary carcinoma, the immunologic function of tumor-bearing mice is had to stronger raising effect simultaneously; To mice people Transplanted Gastric Carcinoma 5-FU, chemotherapy has better potentiation to embodiment 61 samples; Embodiment 61 samples have good protective effect to the caused bone marrow depression of 5-FU chemotherapy, therefore think that embodiment 61 samples have good antitumor action.
Two, the experimentation to radiation damage protective effect
1. experiment material
1.1 animals: Kunming mouse, 18～22g, clean level, purchased from Hunan Si Laike Jing Da laboratory animal company limited, the quality certification number: SCXK(Hunan) 2009-0004.
1.2 medicines and reagent: embodiment, 61 samples are provided by Jiangzhong Pharmaceutical Co., Ltd., are mixed with every milliliter of medicinal liquid 0.286g crude drug that is equivalent to embodiment 61 samples with front with sterile purified water; Ring phosphinylidyne (CTX, Hualian Pharmaceutical Co., Ltd., Shanghai), sheep red blood cell (SRBC) (SRBC, Guangzhou Qi Yun Bioisystech Co., Ltd); SOD detection kit (Bioengineering Research Institute is built up in Nanjing).
1.3 key instrument NanoVue trace albumin nucleic acid instrument (GE company); AL204 electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd.); XT-2000i automated blood analysis (Japanese Sysmex company); ELx800 type microplate reader (BioTek, the U.S.).
2. method
2.1 animal groupings: be divided at random 5 groups according to body weight, 12 every group.Set up blank group, model control group, the basic, normal, high dosage group of embodiment 61 sample.
2.2 dosage designs: everyone recommended intake every day of embodiment 61 samples is: 20g/70kg body weight.Extrapolating thus mice daily intake is: low dose group, 1.43g crude drug/kg; Middle dosage group, 2.86g crude drug/kg; High dose group, 5.72g crude drug/kg, is equivalent to respectively 5,10 and 20 times of people's daily intake.
2.3 impacts of embodiment 61 samples on cobalt source radiation murine peripheral leukocytes quantity: before experiment, each group mouse orbit is got blood 20 μ L, and blood analyser calculates every murine interleukin quantity.Blank group and model group gavage distilled water, the each treated animal gavage of embodiment 61 sample gives corresponding test solution 10mL/kg body weight, every day 1 time, continuous 30 days.Except blank group, all animals carried out in the 16th day
60co irradiates, and irradiation dose is 5Gy.Each group mice the 3rd day, the 14th day eye socket after irradiation got blood 20 μ L, and blood analyser calculates every murine interleukin number.
2.4 embodiment 61 samples affect cobalt source radiation murine bone marrow nucleated cell number, Quantity of DNA and bone marrow cell micronucleus number: grouping and administration same 2.3.Except blank group, all animals all carried out in the 27th day
60co irradiates, irradiation dose is 5Gy, each group mice the 3rd day de-cervical vertebra after irradiation put to death animal, separate femur, syringe is drawn Hank ' the s liquid of certain volume, go out the whole medullary cells in femur, under mirror, count the number of nucleated cells in every mL bone marrow cell suspension, another extracting marrow cell liquid is measured in 260nm place with ultraviolet spectrophotometer, calculate DNA content, the another breast bone that takes out, extrude bone marrow fluid with mosquito forceps, smear, after natural drying, 15min dyes in Giemsa dye liquor, after distilled water flushing, dry, under mirror, count micronucleus cell number in 1000 polychromatic erythrocytes.
The impact of 2.5 embodiment 61 samples on cobalt source radiation murine serum regulating liver-QI tissue SOD activity: grouping and administration same 2.3.Except blank group, all animals all carried out in the 23rd day
60co irradiates, and irradiation dose is 7Gy, respectively organizes mice and within the 7th day after irradiation, gets blood separation of serum, and de-cervical vertebra is put to death animal thereafter, gets liver, and SOD test kit detects each group of mice serum and hepatic tissue SOD activity.
2.6 impacts of embodiment 61 samples on cobalt source radiation murine serum hemolysin: grouping and administration same 2.3.Except blank group, all animals all carried out in the 15th day
60co irradiates, and irradiation dose is 2Gy, and experiment finishes first 4 days, every animal lumbar injection 2%(v/v) SRBC suspension carries out immunity, and 24h after last administration, plucks eyeball and gets blood, and separation of serum carries out the mensuration of serum hemolysin.
HC50=(sample OD value ÷ SRBC HD50 OD value)/extension rate
2.7 impacts of embodiment 61 samples on the cobalt-60 radiosterilize mouse survival time: grouping is with administration with 2.3, and every treated animal number is 20.D1-d7 days, is placed in one by one self-control ray by mice and exposes in box, uses
60co irradiates, every day 1 time, and each absorbed dose 2.0Gy, total absorbed dose is 14Gy.Mean survival time after observation more each treated animal exposure in 30d, and record the general condition of animal.
2.8 statistical method: experimental data represents with `X ± S, adopt SPSS12.0 one factor analysis of variance, the difference between more blank group, model control group, embodiment 61 sample sets.
3. result
3.1 impacts of embodiment 61 samples on cobalt source radiation murine peripheral leukocytes quantity
The results are shown in Table 6.With the comparison of blank group, model control group the 3rd, 14 days WBC values after irradiation obviously decline, compare with model control group, the middle and high dosage group of embodiment 61 sample mice the 3rd, 14 days WBC value values after irradiation obviously raise, and prompting embodiment 61 sample prevention administrations have the effect that the radiation-induced WBC in cobalt source reduces that suppresses.
The impact of 3.2 embodiment 61 samples on cobalt source radiation murine bone marrow nucleated cell number, Quantity of DNA and bone marrow cell micronucleus rate
The results are shown in Table 7.Compared with blank group, model control group mouse bone marrow cells number of nucleated cells obviously reduces, and Quantity of DNA obviously reduces, and bone marrow cell micronucleus rate obviously increases; With model control group comparison, the middle and high dosage group of embodiment 61 sample mouse bone marrow cells number of nucleated cells obviously increases, Quantity of DNA obviously raises, and bone marrow cell micronucleus rate obviously declines, and prompting embodiment 61 samples have the bone marrow injury effect due to good antagonism cobalt-60 radiosterilize.
The impact of 3.3 embodiment 61 samples on cobalt source radiation murine serum regulating liver-QI tissue SOD activity
The results are shown in Table 8, compared with blank group, model group mice serum regulating liver-QI tissue SOD is active obviously to reduce; With model group comparison, the middle and high dosage group of embodiment 61 sample mice serum regulating liver-QI tissue SOD is active obviously to raise, and prompting embodiment 61 samples can resist the impaired of body redox reaction due to irradiation preferably.
3.4 impacts of embodiment 61 samples on hemolysin level in radiation murine blood
Result of the test is in table 9: compared with blank group, model control group mice serum hemolysin level obviously reduces; With model control group comparison, the middle and high dosage group of embodiment 61 sample mice serum hemolysin level obviously raises, and prompting embodiment 61 samples can resist the damage of the body immune system due to irradiation preferably.
3.5 impacts of embodiment 61 samples on the cobalt-60 radiosterilize mouse survival time
The results are shown in Table 10, mice warp
60after Co Continuous irradiation, there is inappetence, body weight significantly alleviates, roll up and movable minimizing, there is the symptom such as loose stool and subcutaneous hemorrhage in Some Animals, most animals, in 7～14d death, is giving after embodiment 61 samples, the survival rate of laboratory animal significantly increases, and mean survival time is significant prolongation also.
4. conclusion
Show through animal experiment study: the embodiment 61 samples cobalt-60 radiosterilize mice periphery WBC value that can obviously raise; And mouse bone marrow cells number of nucleated cells and Quantity of DNA after the cobalt-60 radiosterilize that can raise, obviously reduce bone marrow cell micronucleus rate; The embodiment 61 samples cobalt-60 radiosterilize mice serum regulating liver-QI tissue SOD activity that can raise, mice serum hemolysin level after the irradiation that obviously raises, in addition, survival period and survival ratio that embodiment 61 samples can significant prolongation irradiation mice.
Therefore think, embodiment 61 samples have significant protective effect to the caused body injury of radiotherapy, have protective effect aspect the infringement of the immune system that especially causes at irradiation, body redox reaction and bone marrow damage.
Three, the experimentation of antagonism chemotherapy damage
1. experiment material
1.1 animals: Kunming mouse, 18～22g, clean level, purchased from Hunan Si Laike Jing Da laboratory animal company limited, the quality certification number: SCXK(Hunan) 2009-0004.
1.2 medicines and reagent: embodiment, 61 samples are provided by Jiangzhong Pharmaceutical Co., Ltd., are mixed with every milliliter of medicinal liquid 0.286g crude drug that is equivalent to embodiment 61 samples with front with sterile purified water; Sheep red blood cell (SRBC) (SRBC, Guangzhou Qi Yun Bioisystech Co., Ltd); SOD detection kit (Bioengineering Research Institute is built up in Nanjing).
1.3 key instrument NanoVue trace albumin nucleic acid instrument (GE company); AL204 electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd.); XT-2000i automated blood analysis (Japanese Sysmex company); ELx800 type microplate reader (BioTek, the U.S.).
2. method
2.1 animal groupings: be divided at random 5 groups according to body weight, 12 every group, set up blank group, model control group, the basic, normal, high dosage group of embodiment 61 sample.
2.2 dosage designs: everyone recommended intake every day of embodiment 61 samples is: 20g crude drug/70kg body weight, extrapolate thus mice daily intake and be: low dose group, 1.43g crude drug/kg; Middle dosage group, 2.86g crude drug/kg; High dose group, 5.72g crude drug/kg, is equivalent to respectively 5,10 and 20 times of people's daily intake.
2.3 impacts of embodiment 61 samples on CTX induced mice peripheral leukocytes: before experiment, each group mouse orbit is got blood 20 μ L, blood analyser calculates every murine interleukin quantity, blank group and model group gavage distilled water, the each treated animal gavage of embodiment 61 sample gives corresponding test solution 10mL/kg body weight, every day 1 time, continuous 30 days.Except blank group, all animals started lumbar injection CTX40mg/kg in the 21st day, once a day, respectively organized mice after last administration 24 hours, and eye socket is got blood 20 μ L, and blood analyser detects mouse peripheral blood leukocyte (WBC).
2.4 embodiment 61 samples affect CTX induced mice thymus coefficient, spleen index, bone marrow nucleated cell number, Quantity of DNA and bone marrow cell micronucleus number: grouping and administration same 2.3.Each group mice 24 hours de-cervical vertebras after last administration are put to death animal, get thymus, spleen is weighed, calculate thymus coefficient and index and spleen index, separate femur, syringe is drawn Hank ' the s liquid of certain volume, go out the whole medullary cells in femur, under mirror, count the number of nucleated cells in every mL bone marrow cell suspension.Another extracting marrow cell liquid is measured in 260nm place with ultraviolet spectrophotometer, calculates DNA content.The another breastbone that takes out, extrudes bone marrow fluid with mosquito forceps, smear, and after natural drying, the 15min that dyes in Giemsa dye liquor, dries after distilled water flushing.Under mirror, count micronucleus cell number in 1000 polychromatic erythrocytes.
The impact of 2.5 embodiment 61 samples on CTX induced mice serum regulating liver-QI tissue SOD activity: grouping and administration same 2.3.Each group mice after last administration 24 hours, gets blood separation of serum, and de-cervical vertebra is put to death animal thereafter, gets liver, and SOD test kit detects each group of mice serum and hepatic tissue SOD activity.
2.6 impacts of embodiment 61 samples on CTX induced mice serum hemolysin: grouping and administration same 2.3.Except experiment finishes first 4 days, every animal lumbar injection 2%(v/v) SRBC suspension carries out immunity, and 24h after last administration, plucks eyeball and gets blood, and separation of serum carries out the mensuration of hemolysin.
HC50=(sample OD value ÷ SRBC HD50 OD value)/extension rate
2.7 statistical method: experimental data represents with `X ± S, adopt SPSS12.0 one factor analysis of variance, the difference between more blank group, model control group, embodiment 61 sample sets.
3. result
3.1 impacts of embodiment 61 samples on CTX induced mice peripheral blood cells
The results are shown in Table 11: with the comparison of blank group, model control group WBC value obviously declines.Compare with model control group, the basic, normal, high dosage group of embodiment 61 sample mice WBC value obviously raises, the effect that the WBC that prompting embodiment 61 sample prevention administrations have inhibition CTX to cause reduces.
The impact of 3.2 embodiment 61 samples on CTX induced mice bone marrow nucleated cell number, Quantity of DNA and bone marrow cell micronucleus rate
The results are shown in Table 12: compared with blank group, model control group mouse bone marrow cells number of nucleated cells obviously reduces, and Quantity of DNA obviously reduces, and bone marrow cell micronucleus rate obviously increases; With model control group comparison, the middle and high dosage group of embodiment 61 sample mouse bone marrow cells number of nucleated cells obviously increases, Quantity of DNA obviously raises, and bone marrow cell micronucleus rate obviously declines, and prompting embodiment 61 samples have the bone marrow injury effect due to good antagonism CTX.
The impact of 3.3 embodiment 61 samples on CTX induced mice serum regulating liver-QI tissue SOD activity
The results are shown in Table 13: compared with blank group, model group mice serum regulating liver-QI tissue SOD is active obviously to reduce; With model group comparison, the middle and high dosage group of embodiment 61 sample mice serum regulating liver-QI tissue SOD is active obviously to raise, and prompting embodiment 61 samples can resist the impaired of body redox reaction due to CTX preferably.
The impact of 3.4 embodiment 61 samples on CTX induced mice thymus coefficient, spleen index and serum hemolysin
Result of the test is in table 14: compared with blank group, model control group mice serum hemolysin level obviously reduces; With model control group comparison, the middle and high dosage group of embodiment 61 sample mice serum hemolysin level obviously raises, and prompting embodiment 61 samples can resist the damage of the body immune system due to CTX preferably.
4. conclusion
Show through animal experiment study: the embodiment 61 samples CTX chemotherapy induced mice periphery WBC value that can obviously raise; And mouse bone marrow cells number of nucleated cells and Quantity of DNA after the CTX chemotherapy that can raise, obviously reduce bone marrow cell micronucleus rate; The embodiment 61 samples mice serum regulating liver-QI tissue SOD activity after CTX chemotherapy that can raise, mouse thymus coefficient, index and spleen index and serum hemolysin after the CTX chemotherapy that obviously raises.
Therefore think, embodiment 61 samples have significant protective effect to the caused body injury of chemotherapy, have protective effect aspect the infringement of the immune system that especially causes in chemotherapy, body redox reaction and bone marrow damage.
Four, embodiment 61 samples strengthen the report of immune effect animal experiment
1. experiment material
1.1 animals: C57BL/6 mice, body weight 18-22g, purchased from Beijing Vital River Experimental Animals Technology Co., Ltd., quality certification SCXK(capital) 2006-009.
1.2 medicines and reagent: embodiment, 61 samples are provided by Jiangzhong Pharmaceutical Co., Ltd., are mixed with every milliliter of medicinal liquid 0.286g crude drug that is equivalent to embodiment 61 samples with front with sterile purified water; Concanavalin A, Con A (ConA, Sigma company), MTT(Sigma company), dinitrofluorobenzene (DNFB, Chengdu Gracia chemical technology company limited), Mianyang erythrocyte (SRBC, Guangzhou Qi Yun Bioisystech Co., Ltd), hemoglobin detection kit (Bioengineering Research Institute is built up in Nanjing), india ink (Qingdao Hai Bo Bioisystech Co., Ltd).
1.3 key instrument NanoVue trace albumin nucleic acid instrument (GE company); AL204 electronic balance (Mettler-Toledo Instrument (Shanghai) Co., Ltd.); ELx800 type microplate reader (BioTek, the U.S.).
2. method
2.1 animal groupings: be divided at random 4 groups according to body weight, 12 every group.Set up blank group, the basic, normal, high dosage group of embodiment 61 sample.
2.2 dosage designs: everyone recommended intake every day of embodiment 61 samples is: 20g/70kg body weight.Extrapolating thus mice daily intake is: low dose group, 1.43g crude drug/kg; Middle dosage group, 2.86g crude drug/kg; High dose group, 5.72g crude drug/kg, is equivalent to respectively 5,10 and 20 times of people's daily intake, the basic, normal, high dosage group of embodiment 61 sample respectively gavage gives the medicinal liquid 10ml/kg of respective concentration, once a day, successive administration 30 days, the capacity distilled water such as blank group gavage.
2.3 impacts of embodiment 61 samples on mouse lymphocyte conversion ratio: after last administration 24 hours, de-cervical vertebra is put to death mice, under aseptic condition, get mouse spleen, make single cell suspension, after splitting erythrocyte, adjust splenocyte density with the RPMI-1640 culture fluid containing 10% calf serum, make 5 × 10
6/ ml mouse boosting cell suspension, adds every a splenocyte suspension respectively in two 96 orifice plates, and wherein a hole adds 75 μ l ConA, and another hole adds equivalent culture medium in contrast, and above-mentioned each sample repeats 3 multiple holes, and 96 orifice plates are placed at 37 DEG C 5% CO
2in incubator, cultivate 72h, cultivate and finish front 4h, every hole adds MTT (5mg/ml) 50 μ l, continues to cultivate.After cultivation finishes, abandon supernatant, every hole adds 150 μ l acid isopropyl alcohol, mixes, and measures the optical density at 570nm wavelength place by microplate reader, calculates splenocyte turn over number.
Splenocyte turn over number=ConA stimulation hole OD value-stimulate hole OD value without ConA
The impact of the mice delayed allergy of 2.4 embodiment 61 samples on DNFB induction: administration finishes first 3 days, each group adopts barium sulfide that every Mus skin of abdomen is lost hair or feathers, scope is 3 × 3cm, is applied in unhairing place with the DNFB acetone Oleum Sesami solution 50 μ l of 10mg/ml.After last administration 1h, 10 μ lDNFB acetone Oleum Sesami solution are evenly applied in to mouse right ear and attack.After 24h, the weight of electronic balance weighing left and right two ears, and calculate auricle swelling degree.Swelling=auris dextra weight-left ear weight
2.5 impacts of embodiment 61 samples on mouse boosting cell antibody forming capacity: before experiment finishes the 4th day, except Normal group, every mouse peritoneal injection sheep red blood cell (SRBC) (SRBC) 1 × 10
8individual, experiment finishes the same day, puts to death mice, and the aseptic splenocyte suspension of preparing, adjusts its cell concentration as 5 × 10 taking RPMI1640 culture fluid
6/ ml is poured on the slide of brushing agarose thin layer, makes parallel plate, after agar solidifies, slide level is buckled and is placed on horse, puts into CO
2in incubator, hatch 1.5h, then add complement (the fresh guinea pig serum of 1:8), continue incubation 1.5h, counting hemolysis plaque number.
2.6 impacts of embodiment 61 samples on mice serum hemolysin: experiment finishes first 4 days, except Normal group, every animal lumbar injection 2%(v/v) SRBC suspension carries out immunity.24h after last administration, plucks eyeball and gets blood, separation of serum, and according to State Food and Drug Administration, " function of health food is learned assessment process and method of inspection specification ", the method for 2003 editions is measured HC50, HC50=(sample OD value ÷ SRBC HD50 OD value)/extension rate
2.7 impacts of embodiment 61 samples on mice carbon clearance index: take Mouse Weight after last gavage 1h, with the india ink of 3.5 times of dilutions of every Mus 10g body weight 0.1ml tail vein injection, after injection 2, the 10min posterior orbit 20 μ l that take a blood sample are dissolved in 2ml mass fraction 0.1% Na
2cO
3in solution, after shaking up, with Na
2cO
3solution does blank, measures absorbance in ultraviolet spectrophotometer 600nm wavelength place, to clean up relatively mononuclear phagocyte phagocytic activity of index:
Clean up index K=(lgOD
1-lgOD
2)/(t
2-t
1)
OD
1, OD
2be respectively at time t
1, t
2time institute's blood sampling absorbance; t
2-t
1for getting the time difference of two blood samples.
Last is got after blood, and de-mice neck is put to death, and claims mice to get Mouse Weight, and takes out liver, spleen, thymus, weighs, and with following formula calculating Thymus and Spleen index, and calculates and engulfs factor alpha:
Thymus index=thymus quality (mg)/body weight (g)
Index and spleen index=spleen weight (mg)/body weight (g)
Engulf factor alpha=body weight × K
1/3/ (spleen weight+liver is heavy)
2.8 impacts of embodiment 61 samples on Phagocytosis By The Peritoneal Macrophages In Mice: after last gavage, each group mouse peritoneal is injected 20% chicken red blood cell (CRBC) 1ml interval 30min, animal is put to death in cervical vertebra dislocation, use normal saline flushing abdominal cavity, then sucking-off abdominal cavity liquid 1ml, drops in respectively on two slides, hatch 30min for 37 DEG C, slide is with after the dyeing of Giemsa-phosphate buffer after cleaning, fix, drying, and microscopy counting, calculates phagocytic percentage and phagocytic index.
Phagocytic rate (%)=(engulfing macrophage number/100 macrophage of chicken red blood cell) × 100
Chicken red blood cell number/100 macrophage of phagocytic index=engulfed
The impact of 2.9 embodiment 61 samples on mice natural killer cell (NK) activity: after last administration 24h, extracting spleen cell under aseptic condition, makes splenocyte suspension, adjusting suspension cell concentration is 2 × 10
7/ ml, getting concentration is 2 × 10
5the each 100 μ l of the YAC-1 target cell of/ml and effector lymphocyte (effect target is than 50:1) add U-shaped 96 well culture plates.Target cell Spontaneous release hole adds target cell and the each 100 μ l of culture fluid, and the maximum release aperture of target cell adds target cell and the each 100 μ l of 1%NP40, and above-mentioned every 3 multiple holes of all establishing, in 37 DEG C, 5%CO
2in incubator, cultivate 20 h, then by 96 well culture plates with the centrifugal 5min of 1500r/min, draw at the bottom of supernatant 100 μ l horizontalizations in 96 well culture plates in every hole, add LDH substrate liquid 100 μ l reaction 3min simultaneously, every hole adds 1mol/L HCl 30 μ l, and full-automatic microplate reader 490nm place measures optical density value.
2.10 statistical method: experimental data represents with `X ± S, adopt SPSS12.0 one factor analysis of variance, the difference between more blank group, embodiment 61 sample sets.
3. result
The impact of the lymhocyte transformation rate of 3.1 embodiment 61 samples on ConA induction
The results are shown in Table 15: with the comparison of blank group, the middle and high dosage group of embodiment 61 sample mouse spleen lymphocyte transforms OD difference and obviously raises, prompting embodiment 61 samples transform to mouse spleen lymphocyte the effect of being significantly improved.
The impact of the mice delayed allergy of 3.2 embodiment 61 samples on DNFB induction
The results are shown in Table 16: with the comparison of blank group, the middle and high dosage group of embodiment 61 sample mice ear rate obviously raises, prompting embodiment 61 samples have the effect that promotes delayed allergy.
3.3 impacts of embodiment 61 samples on mouse boosting cell antibody forming capacity
The results are shown in Table 17 compared with blank group, the middle and high dosage group of embodiment 61 sample mice hemolysis plaque number obviously increases, and prompting embodiment 61 samples can better improve mouse boosting cell antibody forming capacity.
3.4 impacts of embodiment 61 samples on mice serum Hemolysin formation
The results are shown in Table 18 compared with blank group, the middle and high dosage group of embodiment 61 sample mice serum Hemolysin formation obviously increases.
3.5 impacts of embodiment 61 samples on mice carbon clearance index
The results are shown in Table 19 compared with blank group, the middle and high dosage group of embodiment 61 sample mouse thymus coefficient, spleen index obviously raise, and can improve phagocytic index and engulf coefficient, prompting embodiment 61 samples can better promote the monocytes/macrophages carbon of mice to clean up the effect of function.
3.6 impacts of embodiment 61 samples on Phagocytosis By The Peritoneal Macrophages In Mice
The results are shown in Table 20: compared with blank group, embodiment 61 sample middle and high dosage group mice chicken red blood cell phagocytic rate and phagocytic index are all significantly increased, prompting embodiment 61 samples can better promote Phagocytosis By The Peritoneal Macrophages In Mice.
3.7 impacts of embodiment 61 samples on NK cells in mice activity
The results are shown in Table 21 compared with blank group, the middle and high dosage group of embodiment 61 sample NK cells in mice is active obviously to be increased.
4. conclusion
Can find out from above zoopery, significantly mouse lymphocyte conversion ratio of embodiment 61 samples, strengthen delayed allergy ability, improving mouse boosting cell antibody forms and serum hemolysin generative capacity, also can increase mice carbon clearance ability and peritoneal macrophage and engulf chicken red blood cell ability, also can increase the killing activity of NK cells in mice to target cell simultaneously, therefore, embodiment 61 samples have enhancing body cellular immunization and humoral immune function, can strengthen the cell killing activity of the huge phagocytic function of biting of monocytes/macrophages and abdominal cavity and NK cell simultaneously.
Embodiment 62
Turtle peptide 150kg, soybean peptide 30kg, ocean fish peptide 20kg add turtle peptide, soybean peptide, ocean fish peptide to carry out batch mixing in blender, use aluminum composite membrane bag to pack, 10 grams every bag.
Embodiment 62 has carried out following quality control to embodiment 61 samples:
One, total peptide content is measured
1. method summary
Low-molecular-weight protein hydrolysate (comprising oligopeptide and free amino acid) dissolves in trichloroacetic acid solution; The protein of high molecular easily precipitates in trichloroacetic acid solution.Sample through trichloroacetic acid solution dissolve after, separable removal high molecular weight protein material.In alkaline solution, peptide bond and Cu2+ chelating, form peptide-copper composition, and this complex makes the phosphomolybdic acid reduction of phenol reagent, produces blue compound, and this blueness compound is directly proportional to oligopeptide content at the absorbance at 650nm place.Measure peptide total amount with this.
2. analytical procedure
2.1 reagent
Ultra-pure water; 19% trichloroacetic acid; 4% sodium carbonate; 0.8% sodium hydroxide solution; 1% copper sulfate; 2% potassium sodium tartrate solution; Phenol reagent;
Alkaline copper test solution: get 2% potassium sodium tartrate solution and the each 0.5mL of 1% copper-bath, 4% sodium carbonate liquor and the each 25mL of 0.8% sodium hydroxide solution, shake up, and to obtain final product.This liquid should be prepared before use.
The preparation of 2.2 need testing solutions
Precision takes embodiment 62 sample 1g, put in 25ml volumetric flask, add water appropriate, jolting makes to dissolve, if desired can supersound process, add water and be settled to 25ml, centrifugal 15min(4000r/min), precision measures in the volumetric flask of supernatant 2ml to 10ml, add 19% trichloroacetic acid to scale, shake up, leave standstill 10min, centrifugal 15min(4000r/min).Precision measures in supernatant 1mL to 100ml measuring bottle, is diluted with water to scale, shakes up, and is need testing solution.
The preparation of 2.2 bovine serum albumin standard solution
It is appropriate that precision takes bovine serum albumin standard substance (V component), puts in measuring bottle, with water dissolution, is diluted to scale, shakes up, and making concentration is 0.1mg/mL, before use preparation.
2.3 assay method
Precision pipettes test sample and prepares liquid 1mL, puts in 10mL tool plug test tube, adds alkaline copper solution 5mL, shakes up.Room temperature is placed after 10min, add fast phenol reagent 0.5mL, fully shake up, put in 30 DEG C of water-baths constant temperature 30 minutes, take out, let cool, measure absorbance in wavelength 650nm place, precision pipettes standard protein solution 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL and is placed in respectively test tube, add water to 1mL, from " adding alkaline copper solution 5mL ", with method operation, retinue blank.With the corresponding absorbance of standard substance concentration, ask equation of linear regression, by the test sample absorbance substitution regression equation recording, calculate test sample content.
3. measurement result
Embodiment 62 5 duplicate samples are measured respectively total peptide content, the results are shown in Table 22.
Two, relative molecular mass is less than the protein hydrolysate proportion (high performance gel filtration chromatography) of 10000u
1. method summary
Employing high performance gel filtration chromatography is measured.Taking porous filler as immobile phase, difference according to sample component molecular volume size separates, under the uv absorption wavelength 220nm of peptide bond condition, detect, use gel chromatography to measure the exclusive data process software (being GPC software) that relative molecular mass distributes, chromatogram and data thereof are processed, calculate relative molecular mass size and the distribution of protein hydrolysate, and then obtain the protein hydrolysate proportion that relative molecular mass is less than 10000u.
2. analytical procedure
2.1 reagent
Acetonitrile: chromatographically pure; Trifluoracetic acid: analytical pure; Water: ultra-pure water or redistilled water.
Relative molecular mass calibration trace standard substance used: cytochrome C (cyyochrome, Mw12500); Press down phthalein enzyme (aprotinin, Mw6500); Bacillus enzyme (bacitracin, Mw1450); Aminoacetic acid-aminoacetic acid-tyrosine-arginine (Mw451); Aminoacetic acid-aminoacetic acid-aminoacetic acid (Mw189)
2.2 instrument and equipment
High performance liquid chromatograph: the chromatographic work station of being furnished with UV-detector and containing GPC data processing software; Mobile phase vacuum filtration degasser; Ultrasonic oscillator; Analytical balance: sensibility reciprocal 0.0001g.
2.3 chromatographic conditions and system suitability test
Chromatographic column: TSKgel G2000 SWXL 300mm × 7.8mm or performance therewith close of the same type other are applicable to measure the gel column of proteins and peptides; Mobile phase: acetonitrile: water: trifluoroacetic acid, 20:80:0.1(volume ratio) detection wavelength: UV220nm; Flow velocity: 0.5ml/min; Column temperature: 30 DEG C; Sampling volume: 10 μ l.
For making the requirement of chromatographic system coincidence detection, be defined under above-mentioned chromatographic condition, the post effect of gel chromatographic columns is that theoretical cam curve (N) is calculated and is not less than 5000 by three poly saccharide peptide standard products (aminoacetic acid-aminoacetic acid-aminoacetic acid) peak, and the partition coefficient (Kd) of oligopeptide should be between 0～1.
2.4 relative molecular mass calibration traces are made
Be mixed with respectively the poly saccharide peptide standard product solution of 0.1% above-mentioned different relative molecular masses by mobile phase, with aperture be sample introduction respectively after 0.2 μ m ~ 0.5 μ m politef or nylon filter membrane filter, obtain the chromatogram of series standard product.Logarithm (lgMw) with relative molecular mass is mapped or obtains relative molecular mass calibration trace and equation thereof as linear regression retention time.
2.5 sample preparation
Take sample 20.0mg in 10mL volumetric flask, be settled to scale by mobile phase, sonic oscillation 10min, makes sample fully dissolve and mix, with aperture be after 0.2-0.5 μ m politef or nylon filter membrane filter, upper machine sample introduction.
The calculating of 2.6 relative molecular masses
The sample solution of 2.5 preparations is analyzed under above-mentioned chromatographic condition.Then use GPC data processing software, by calculating in the chromatographic data substitution calibration trace equation of sample, can obtain relative molecular mass and the distribution thereof of protein hydrolysate in sample.Peak area relative percentage sum by areas of peak normalization method calculating relative molecular mass scope below 10000u.
3. measurement result
Get embodiment 62 4 duplicate samples determining molecular weight distributions, measurement result is in table 23.
Above result shows, the molecular weight of embodiment 62 mainly concentrates on below 10000u, and the following material of 10000u molecular weight accounts for more than 95%.
Embodiment 63
Turtle peptide 150Kg, soybean peptide 30kg, ocean fish peptide 20kg, water soluble dietary fiber 15kg, wolfberry fruit extract 2kg, disodiumedetate 0.03kg, citric acid 2.5kg, potassium citrate 1.5kg, maltose alcohol 90kg, sucralose 0.25kg, phosphopeptide caseinate 0.25kg, zinc gluconate 0.04 kg, essence 1.4kg, in preparing tank, add 500L purified water, add successively disodiumedetate, turtle peptide, ocean fish peptide, water soluble dietary fiber, wolfberry fruit extract, citric acid, potassium citrate, maltose alcohol, after stirring is dissolved it, (aperture is 30 μ m to filter, 0.45 μ m) filters, obtain filtrate, by sucralose, phosphopeptide caseinate, after zinc gluconate dissolves by appropriate purified water, join in filtrate, add water to 1000L, stir 15 minutes, add essence, stir again 10 minutes, with filter filtration (sheet material: B-2, aperture: 0.45 μ m), every bottle of 100ml, 105 DEG C of sterilizings 30 minutes, obtain.
The sample of embodiment 63 carries out following quality control experiment:
1. method summary
Low-molecular-weight protein hydrolysate (comprising oligopeptide and free amino acid) dissolves in trichloroacetic acid solution; The protein of high molecular easily precipitates in trichloroacetic acid solution.Sample through trichloroacetic acid solution dissolve after, separable removal high molecular weight protein material.In alkaline solution, peptide bond and Cu2+ chelating, form peptide-copper composition, and this complex makes the phosphomolybdic acid reduction of phenol reagent, produces blue compound, and this blueness compound is directly proportional to oligopeptide content at the absorbance at 650nm place.Measure peptide total amount with this.
2. analytical procedure
2.1 reagent
Ultra-pure water; 19% trichloroacetic acid; 4% sodium carbonate; 0.8% sodium hydroxide solution; 1% copper sulfate; 2% potassium sodium tartrate solution; Phenol reagent;
Alkaline copper test solution: get 2% potassium sodium tartrate solution and the each 0.5mL of 1% copper-bath, 4% sodium carbonate liquor and the each 25mL of 0.8% sodium hydroxide solution, shake up, and to obtain final product.This liquid should be prepared before use.
The preparation of 2.2 need testing solutions
The accurate embodiment 63 sample solution 2ml that draw, put in 25ml volumetric flask, add 19% trichloroacetic acid 8ml, shake up, and leave standstill 10min, add water and are settled to 25ml, centrifugal 15min(4000r/min).Precision measures in supernatant 1mL to 100ml measuring bottle, is diluted with water to scale, shakes up, and is need testing solution.
The preparation of 2.2 bovine serum albumin standard solution
It is appropriate that precision takes bovine serum albumin standard substance (V component), puts in measuring bottle, with water dissolution, is diluted to scale, shakes up, and making concentration is 0.1mg/mL, before use preparation.
2.3 assay method
Precision pipettes test sample and prepares liquid 1mL, puts in 10mL tool plug test tube, adds alkaline copper solution 5mL, shakes up.Room temperature is placed after 10min, add fast phenol reagent 0.5mL, fully shake up, put in 30 DEG C of water-baths constant temperature 30 minutes, take out, let cool, measure absorbance in wavelength 650nm place, precision pipettes standard protein solution 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL and is placed in respectively test tube, add water to 1mL, from " adding alkaline copper solution 5mL ", with method operation, retinue blank.With the corresponding absorbance of standard substance concentration, ask equation of linear regression, by the test sample absorbance substitution regression equation recording, calculate test sample content.
3. measurement result
Embodiment 63 5 duplicate samples are measured respectively total peptide content, the results are shown in Table 24
Embodiment 64
Ocean fish peptide: Hai Shi bio tech ltd, Zhejiang product batch number: 20120318
Turtle peptide: Jiangzhong Pharmaceutical Co., Ltd. provides
The preparation of turtle peptide: Trionyx sinensis Wiegmann, gill and oils and fats, blend, decoct 2h, be cooled to 40 ° of C, add 2.5% ratio neutral protease of the weight of raw material Trionyx sinensis Wiegmann, constant temperature enzymolysis 4h, press again the flavor protease of raw material Trionyx sinensis Wiegmann weight 1%, stir enzymolysis 2h, the enzymolysis solution obtaining is warming up to 100 ° of C, enzyme denaturing 15min, the centrifugal rear filtration of cooling rear enzymolysis solution, supernatant is through thin film concentration, and concentrated solution spraying is dried and obtains turtle peptide;
Take turtle peptide 180kg, ocean fish peptide 50kg, in blender, add turtle peptide, ocean fish peptide to carry out batch mixing, use aluminum aluminum to pack, 10 grams every bag.
The sample of embodiment 64 samples carries out following efficacy test:
One, the experimentation of embodiment 64 samples to radiation damage protective effect
1. experiment material
1.1 animals: with embodiment 61
1.2 medicines and reagent: embodiment, 64 samples are provided by Jiangzhong Pharmaceutical Co., Ltd., are mixed with every milliliter of medicinal liquid 0.329g crude drug that is equivalent to embodiment 64 samples with front with sterile purified water; Sheep red blood cell (SRBC) (SRBC, Guangzhou Qi Yun Bioisystech Co., Ltd); SOD detection kit (Bioengineering Research Institute is built up in Nanjing).
1.3 key instruments are with embodiment 61.
2. method
2.1 animal groupings: be divided at random 5 groups according to body weight, 12 every group.Set up blank group, model control group, the basic, normal, high dosage group of embodiment 64 sample.
2.2 dosage designs: everyone recommended intake every day of embodiment 64 samples is: 23g crude drug/70kg body weight.Extrapolating thus mice daily intake is: low dose group, 1.65g crude drug/kg; Middle dosage group, 3.29g crude drug/kg; High dose group, 6.58g crude drug/kg, is equivalent to respectively 5,10 and 20 times of people's daily intake.
2.3 impacts of embodiment 64 samples on cobalt source radiation murine peripheral leukocytes quantity: before experiment, each group mouse orbit is got blood 20 μ L, and blood analyser calculates every murine interleukin quantity.Blank group and model group gavage distilled water, the each treated animal gavage of embodiment 64 sample gives corresponding test solution 10mL/kg body weight, every day 1 time, continuous 30 days.Except blank group, all animals carried out in the 16th day
60co irradiates, and irradiation dose is 5Gy.Each group mice the 3rd day, the 14th day eye socket after irradiation got blood 20 μ L, and blood analyser calculates every murine interleukin number.
2.4 embodiment 64 samples affect cobalt source radiation murine bone marrow nucleated cell number, Quantity of DNA and bone marrow cell micronucleus number: method is with embodiment 61.
The impact of 2.5 embodiment 64 samples on cobalt source radiation murine serum regulating liver-QI tissue SOD activity: method is with embodiment 61.
2.6 impacts of embodiment 64 samples on cobalt source radiation murine serum hemolysin: method is with embodiment 61.
2.7 impacts of embodiment 64 samples on the cobalt-60 radiosterilize mouse survival time: method is with embodiment 61.
2.8 statistical method: experimental data represents with `X ± S, adopt SPSS12.0 one factor analysis of variance, the difference between more blank group, model control group, embodiment 64 sample sets.
3. result
3.1 impacts of embodiment 64 samples on cobalt source radiation murine peripheral leukocytes quantity
The results are shown in Table 25.With the comparison of blank group, model control group the 3rd, 14 days WBC values after irradiation obviously decline.Compare with model control group, the middle and high dosage group of embodiment 64 sample mice the 3rd, 14 days WBC value values after irradiation obviously raise, and prompting embodiment 64 sample prevention administrations have the effect that the radiation-induced WBC in cobalt source reduces that suppresses.
The impact of 3.2 embodiment 64 samples on cobalt source radiation murine bone marrow nucleated cell number, Quantity of DNA and bone marrow cell micronucleus rate
The results are shown in Table 26: compared with blank group, model control group mouse bone marrow cells number of nucleated cells obviously reduces, and Quantity of DNA obviously reduces, and bone marrow cell micronucleus rate obviously increases; With model control group comparison, the middle and high dosage group of embodiment 64 sample mouse bone marrow cells number of nucleated cells obviously increases, Quantity of DNA obviously raises, and bone marrow cell micronucleus rate obviously declines, and prompting embodiment 64 samples have the bone marrow injury effect due to good antagonism cobalt-60 radiosterilize.
The impact of 3.3 embodiment 64 samples on cobalt source radiation murine serum regulating liver-QI tissue SOD activity
The results are shown in Table 27.Compared with blank group, model group mice serum regulating liver-QI tissue SOD is active obviously to reduce; With model group comparison, the middle and high dosage group of embodiment 64 sample mice serum regulating liver-QI tissue SOD is active obviously to raise, and prompting embodiment 64 samples can resist the impaired of body redox reaction due to irradiation preferably.
3.4 impacts of embodiment 64 samples on hemolysin level in radiation murine blood
Result of the test is in table 28.Compared with blank group, model control group mice serum hemolysin level obviously reduces; With model control group comparison, the middle and high dosage group of embodiment 64 sample mice serum hemolysin level obviously raises, and prompting embodiment 64 samples can resist the damage of the body immune system due to irradiation preferably.
3.5 impacts of embodiment 64 samples on the cobalt-60 radiosterilize mouse survival time
The results are shown in Table 29, mice warp
60after Co Continuous irradiation, occur inappetence, body weight significantly alleviates, and rolls up and movable minimizing, and the symptom such as loose stool and subcutaneous hemorrhage appears in Some Animals.Most animals is in 7～14d death.Giving after embodiment 64 samples, the survival rate of laboratory animal significantly increases, and mean survival time is significant prolongation also.
4. conclusion
Show through animal experiment study: the embodiment 64 samples cobalt-60 radiosterilize mice periphery WBC value that can obviously raise; And mouse bone marrow cells number of nucleated cells and Quantity of DNA after the cobalt-60 radiosterilize that can raise, obviously reduce bone marrow cell micronucleus rate; The embodiment 64 samples cobalt-60 radiosterilize mice serum regulating liver-QI tissue SOD activity that can raise, mice serum hemolysin level after the irradiation that obviously raises.In addition, embodiment 64 samples can significant prolongation irradiation mice survival period and survival ratio.
Therefore think, embodiment 64 samples have significant protective effect to the caused body injury of radiotherapy, have protective effect aspect the infringement of the immune system that especially causes at irradiation, body redox reaction and bone marrow damage.
Two, the experimentation of the anti-chemotherapy damage of embodiment 64 sample
1. experiment material
1.1 animals: Kunming mouse, 18～22g, clean level, purchased from Hunan Si Laike Jing Da laboratory animal company limited, the quality certification number: SCXK(Hunan) 2009-0004.
1.2 medicines and reagent: embodiment, 64 samples are provided by Jiangzhong Pharmaceutical Co., Ltd., are mixed with every milliliter of medicinal liquid 0.329g crude drug that is equivalent to embodiment 64 samples with front with sterile purified water; Ring phosphinylidyne (CTX, Hualian Pharmaceutical Co., Ltd., Shanghai), sheep red blood cell (SRBC) (SRBC, Guangzhou Qi Yun Bioisystech Co., Ltd); SOD detection kit (Bioengineering Research Institute is built up in Nanjing).
1.3 key instrument methods are with embodiment 61.
2. method
2.1 animal groupings: be divided at random 5 groups according to body weight, 12 every group.Set up blank group, model control group, the basic, normal, high dosage group of embodiment 64 sample.
2.2 dosage designs: embodiment 64 everyone recommended intakes every day are: 23g crude drug/70kg body weight.Extrapolating thus mice daily intake is: low dose group, 1.65g crude drug/kg; Middle dosage group, 3.29g crude drug/kg; High dose group, 6.58g crude drug/kg, is equivalent to respectively 5,10 and 20 times of people's daily intake.
2.3 impacts of embodiment 64 samples on CTX induced mice peripheral leukocytes: method is with embodiment 61.
2.4 embodiment 64 samples affect CTX induced mice thymus coefficient, spleen index, bone marrow nucleated cell number, Quantity of DNA and bone marrow cell micronucleus number: method is with embodiment 61.
The impact of 2.5 embodiment 64 samples on CTX induced mice serum regulating liver-QI tissue SOD activity: method is with embodiment 61.
2.6 impacts of embodiment 64 samples on CTX induced mice serum hemolysin: method is with embodiment 61.
3. result
3.1 impacts of embodiment 64 samples on CTX induced mice peripheral blood cells
The results are shown in Table 30.With the comparison of blank group, model control group WBC value obviously declines.Compare with model control group, the basic, normal, high dosage group of embodiment 64 sample mice WBC value obviously raises, the effect that the WBC that prompting embodiment 64 sample prevention administrations have inhibition CTX to cause reduces.
The impact of 3.2 embodiment 64 samples on CTX induced mice bone marrow nucleated cell number, Quantity of DNA and bone marrow cell micronucleus rate
The results are shown in Table 31.Compared with blank group, model control group mouse bone marrow cells number of nucleated cells obviously reduces, and Quantity of DNA obviously reduces, and bone marrow cell micronucleus rate obviously increases; With model control group comparison, the middle and high dosage group of embodiment 64 sample mouse bone marrow cells number of nucleated cells obviously increases, Quantity of DNA obviously raises, and bone marrow cell micronucleus rate obviously declines, and prompting embodiment 64 samples have the bone marrow injury effect due to good antagonism CTX.
The impact of 3.3 embodiment 64 samples on CTX induced mice serum regulating liver-QI tissue SOD activity
The results are shown in Table 32.Compared with blank group, model group mice serum regulating liver-QI tissue SOD is active obviously to reduce; With model group comparison, the middle and high dosage group of embodiment 64 sample mice serum regulating liver-QI tissue SOD is active obviously to raise, and prompting embodiment 64 samples can resist the impaired of body redox reaction due to CTX preferably.
The impact of 3.4 embodiment 64 samples on CTX induced mice thymus coefficient, spleen index and serum hemolysin
Result of the test is in table 33.Compared with blank group, model control group mice serum hemolysin level obviously reduces; With model control group comparison, the middle and high dosage group of embodiment 64 sample mice serum hemolysin level obviously raises, and prompting embodiment 64 samples can resist the damage of the body immune system due to CTX preferably.
4. conclusion
Show through animal experiment study: the embodiment 64 samples CTX chemotherapy induced mice periphery WBC value that can obviously raise; And mouse bone marrow cells number of nucleated cells and Quantity of DNA after the CTX chemotherapy that can raise, obviously reduce bone marrow cell micronucleus rate; The embodiment 64 samples mice serum regulating liver-QI tissue SOD activity after CTX chemotherapy that can raise, mouse thymus coefficient, index and spleen index and serum hemolysin after the CTX chemotherapy that obviously raises.
Therefore think, embodiment 64 samples have significant protective effect to the caused body injury of chemotherapy, have protective effect aspect the infringement of the immune system that especially causes in chemotherapy, body redox reaction and bone marrow damage.
Three, embodiment 64 sample antitumor action animal experiment reports
1. experiment material
1.1 animals: with embodiment 61.
1.2 medicines and reagent: embodiment, 64 samples are provided by Jiangzhong Pharmaceutical Co., Ltd., are mixed with every milliliter of medicinal liquid 0.329g crude drug that is equivalent to embodiment 64 samples with front with sterile purified water.5-fluorouracil injection (5-FU, Shandong Pharmaceutical all together), concanavalin A, Con A (ConA, Sigma company), MTT(Sigma company), IL-2, TNF-α ELISA detection kit be all purchased from ExCell company.
1.3 key instruments are with embodiment 61.
2. method
2.1 dosage designs: the sample of embodiment 64, everyone is at recommended intake every day: 23g crude drug/70kg body weight.Extrapolating thus mice daily intake is: low dose group, 1.65g crude drug/kg; Middle dosage group, 3.29g crude drug/kg; High dose group, 6.58g crude drug/kg, is equivalent to respectively 5,10 and 20 times of people's daily intake.
The tumor-inhibiting action of 2.2 embodiment 64 samples to murine sarcoma S180: the mice cervical vertebra dislocation of 8 days after intraperitoneal inoculation Mouse sarcoma S180 cells is put to death, take out ascites under aseptic condition, with normal saline 1:2 dilution.Mice is divided into 4 groups at random by body weight, 10 every group, sets up model control group, the basic, normal, high dosage group of embodiment 64 sample.Give every mouse armpit subcutaneous vaccination tumor liquid 0.2mL, next day after inoculation, the basic, normal, high dosage of embodiment 64 sample is given the embodiment 64 sample medicinal liquids of pressing respectively 1.65g crude drug/kg body weight, 3.29g crude drug/kg body weight, the basic, normal, high concentration of 6.58g crude drug/kg body weight gavage, every day 1 time, successive administration 30 days.After last administration 24 hours, animal was put to death in cervical vertebra dislocation, and point another name tumor weight, calculates inhibition rate of tumor growth (%).Computing formula is: the flat tumor weight of average tumor weight-treatment group is all organized in inhibition rate of tumor growth %=(contrast) the average tumor of ÷ matched group heavy × 100%.
The tumor-inhibiting action of 2.3 embodiment 64 samples to mice transplanted tumor Lewis pulmonary carcinoma: method is with embodiment 61.
2.4 impacts of embodiment 64 samples on tumor-bearing mice immunologic function
2.4.1 the impact of embodiment 64 samples on tumor-bearing mice lymhocyte transformation rate: method is with embodiment 61.
2.4.2 embodiment 64 samples affect method with embodiment 61 to S180 mice serum TNF-α and IL-2 content.
The impact of the potentiation of 2.5 embodiment 64 samples on the chemotherapy of people's Transplanted Gastric Carcinoma mice: mice is in back intradermal vaccination BGC-823 gastric carcinoma cells 1 × 10
7/ only, when gross tumor volume grows to 1 cm
3, under aseptic condition, take off part tumor piece when the left and right, be cut into 1 mm
3the tumor piece of left and right, is inoculated into mouse back with the trocar subcutaneous, forms subcutaneous transplantation tumor.After inoculating 48 h, be divided at random five groups: 1. blank group, only gives equal-volume solvent; 2. 5-FU treatment group, from after inoculation the 28th day, lumbar injection 5-FU, dosage is 25 mg/kg, every day 1 time, continuous 14 days.3. 5-FU+ low dosage embodiment 64 sample sets started to give 5-FU from after inoculation the 28th day, the same 2. group of dosage, and embodiment 64 sample 1.65g every day crude drug/kg, after inoculation the 7th day starts administration, continuous 35 days, every day 1 time; 4. dose delivery example 64 sample sets in 5-FU+ started to give 5-FU from after inoculation the 28th day, the same 2. group of dosage, and embodiment 64 sample 3.290g every day crude drug/kg, after inoculation the 7th day starts administration, continuous 35 days, every day 1 time; 5. 5-FU+ high dose embodiment 64 sample sets started to give 5-FU from after inoculation the 28th day, the same 2. group of dosage, and embodiment 64 sample 6.58g every day crude drug/kg, after inoculation the 7th day starts administration, continuous 35 days, every day 1 time.Above-mentioned each treated animal finishes experiment after 5 weeks, peel off tumor tissues and weigh and calculate tumor control rate.
The impact of the Attenuation of 2.6 embodiment 64 samples on the chemotherapy of people's Transplanted Gastric Carcinoma mice: method is with embodiment 61.
2.7 statistical method: method is with embodiment 61.
3 results
3.1 embodiment 64 samples are to mice S180 and Lewis pulmonary carcinoma tumor-inhibiting action
The results are shown in Table 34.With model control group comparison, the middle and high dosage of embodiment 64 sample has stronger inhibitory action to murine sarcoma S180, and suppression ratio is respectively 33.15% and 39.22%.The middle and high dosage of embodiment 64 sample has stronger inhibitory action to Mice Bearing Lewis pulmonary carcinoma, and suppression ratio is respectively 26.47% and 36.27%.
3.2 embodiment 64 samples on tumor-bearing mice immunologic function impact
3.2.1 the impact of embodiment 64 samples on transplanted tumor S180 mice, Lewis lung cancer in mice splenocyte conversion ratio
The results are shown in Table 35.Compared with model control group, the middle and high dosage of embodiment 64 sample all has stronger raising effect to mice transplanted tumor S180 mice, Lewis lung cancer in mice splenocyte conversion ratio.
3.2.2 the impact of embodiment 64 samples on transplanted tumor S180 mice serum TNF-α and IL-2 content
The results are shown in Table 36.With model control group comparison, the middle and high dosage of embodiment 64 sample all has stronger raising effect to transplanted tumor S180 mice serum TNF-α and IL-2 content.
The potentiation of 3.3 embodiment 64 samples to people's Transplanted Gastric Carcinoma mice 5-FU chemotherapy
The results are shown in Table 37.After embodiment 64 samples and 5-FU therapeutic alliance, the growth of tumor is subject to obvious inhibition, has statistical significance compared with model control group.Compared with the independent treatment group of 5-FU, the tumor weight average of each therapeutic alliance group has reduction in various degree, and tumor growth is subject to further inhibition, and presents dose dependent, but does not occur statistical significance.
The impact of the Attenuation of 3.4 embodiment 64 samples on people's Transplanted Gastric Carcinoma mice 5-FU chemotherapy
The results are shown in Table 38.Tumor animal is after 5-FU successive administration 14 days; there is obvious bone marrow inhibition in animal subject; leukocyte counts and bone marrow DNA content are all significantly lower than matched group; and animal subject is giving after 5 weeks embodiment 64 samples continuously; leukocyte counts and bone marrow DNA content all have rebound significantly, and embodiment 64 samples have obvious bone marrow protective effect.
4. conclusion:
Animal experiment study shows: the middle and high dosage of embodiment 64 sample has stronger inhibitory action to murine sarcoma S180 and mice transplanted tumor Lewis pulmonary carcinoma, the immunologic function of tumor-bearing mice is had to stronger raising effect simultaneously; To mice people Transplanted Gastric Carcinoma 5-FU, chemotherapy has better potentiation to embodiment 64 samples; Embodiment 64 samples have good protective effect to the caused bone marrow depression of 5-FU chemotherapy.Therefore think that embodiment 64 samples have good antitumor action.
Four, the sample of embodiment 64 strengthens the report of immune effect animal experiment
1. experiment material
1.1 animals: with embodiment 61.
1.2 medicines and reagent: embodiment, 64 samples are provided by Jiangzhong Pharmaceutical Co., Ltd., are mixed with every milliliter of medicinal liquid 0.329g crude drug that is equivalent to embodiment 64 samples with front with sterile purified water.Concanavalin A, Con A (ConA, Sigma company), MTT(Sigma company), dinitrofluorobenzene (DNFB, Chengdu Gracia chemical technology company limited), Mianyang erythrocyte (SRBC, Guangzhou Qi Yun Bioisystech Co., Ltd), hemoglobin detection kit (Bioengineering Research Institute is built up in Nanjing), india ink (Qingdao Hai Bo Bioisystech Co., Ltd).
1.3 key instruments: with embodiment 61.
2. method
2.1 animal groupings: be divided at random 4 groups according to body weight, 12 every group.Set up blank group, the basic, normal, high dosage group of embodiment 64 sample.
2.2 dosage designs: executing example 64 everyone recommended intakes every day is: 23g crude drug/70kg body weight.Extrapolating thus mice daily intake is: low dose group, 1.65g crude drug/kg; Middle dosage group, 3.29g crude drug/kg; High dose group, 6.58g crude drug/kg, is equivalent to respectively 5,10 and 20 times of people's daily intake.The basic, normal, high dosage group of embodiment 64 sample respectively gavage gives the medicinal liquid 10ml/kg of respective concentration, once a day, and successive administration 30 days, the capacity distilled water such as blank group gavage.
2.3 impacts of embodiment 64 samples on mouse lymphocyte conversion ratio: method is with embodiment 61.
The impact of the mice delayed allergy of 2.4 embodiment 64 samples on DNFB induction: method is with embodiment 61.
2.5 impacts of embodiment 64 samples on mouse boosting cell antibody forming capacity: method is with embodiment 61.
2.6 impacts of embodiment 64 samples on mice serum hemolysin: method is with embodiment 61.
2.7 impacts of embodiment 64 samples on mice carbon clearance index: method is with embodiment 61.
2.8 impacts of embodiment 64 samples on Phagocytosis By The Peritoneal Macrophages In Mice: method is with embodiment 61.
The impact of 2.9 embodiment 64 samples on mice natural killer cell (NK) activity: method is with embodiment 61.
2.10 statistical method: experimental data represents with `X ± S, adopt SPSS12.0 one factor analysis of variance, the difference between more blank group, embodiment 64 sample sets.
3. result
The impact of the lymhocyte transformation rate of 3.1 embodiment 64 samples on ConA induction
The results are shown in Table 39.With the comparison of blank group, the middle and high dosage group of embodiment 64 sample mouse spleen lymphocyte transforms OD difference and obviously raises, and prompting embodiment 64 samples transform to mouse spleen lymphocyte the effect of being significantly improved.
The impact of the mice delayed allergy of 3.2 embodiment 64 samples on DNFB induction
The results are shown in Table 40.With the comparison of blank group, the middle and high dosage group of embodiment 64 sample mice ear rate obviously raises, and prompting embodiment 64 samples have the effect that promotes delayed allergy.
3.3 impacts of embodiment 64 samples on mouse boosting cell antibody forming capacity
The results are shown in Table 41.Compared with blank group, the middle and high dosage group of embodiment 64 sample mice hemolysis plaque number obviously increases, and prompting embodiment 64 samples can better improve mouse boosting cell antibody forming capacity.
3.4 impacts of embodiment 64 samples on mice serum Hemolysin formation
The results are shown in Table 42.Compared with blank group, the middle and high dosage group of embodiment 64 sample mice serum Hemolysin formation obviously increases.
3.5 impacts of embodiment 64 samples on mice carbon clearance index
The results are shown in Table 43.Compared with blank group, the middle and high dosage group of embodiment 64 sample mouse thymus coefficient, spleen index obviously raise, and can improve phagocytic index and engulf coefficient, prompting embodiment 64 samples can better promote the monocytes/macrophages carbon of mice to clean up the effect of function.
3.6 impacts of embodiment 64 samples on Phagocytosis By The Peritoneal Macrophages In Mice
The results are shown in Table 44.Compared with blank group, embodiment 64 sample middle and high dosage group mice chicken red blood cell phagocytic rate and phagocytic index are all significantly increased, and prompting embodiment 64 samples can better promote Phagocytosis By The Peritoneal Macrophages In Mice.
3.7 impacts of embodiment 64 samples on NK cells in mice activity
The results are shown in Table 45.Compared with blank group, the middle and high dosage group of embodiment 64 sample NK cells in mice is active obviously to be increased.
4. conclusion
Can find out from above zoopery, significantly mouse lymphocyte conversion ratio of embodiment 64 samples, strengthen delayed allergy ability, improving mouse boosting cell antibody forms and serum hemolysin generative capacity, also can increase mice carbon clearance ability and peritoneal macrophage and engulf chicken red blood cell ability, also can increase the killing activity of NK cells in mice to target cell simultaneously.Therefore, embodiment 64 samples have enhancing body cellular immunization and humoral immune function, can strengthen the cell killing activity of the huge phagocytic function of biting of monocytes/macrophages and abdominal cavity and NK cell simultaneously.
In a word, embodiment 64 samples have the effect that strengthens immune function of mice.
Embodiment 65
Turtle peptide 100kg, ocean fish peptide 100kg add turtle peptide, ocean fish peptide to carry out batch mixing in blender, use aluminum aluminum to pack, 10 grams every bag.
Embodiment 66
Turtle peptide 2000kg, ocean fish peptide 1000kg add turtle peptide, ocean fish peptide to carry out batch mixing in blender, use aluminum aluminum to pack, 10 grams every bag.
Embodiment 67
Turtle peptide 100kg, ocean fish peptide 50kg add turtle peptide, ocean fish peptide to carry out batch mixing in blender, use aluminum aluminum to pack, 10 grams every bag.
Embodiment 68
Turtle peptide 180kg, ocean fish peptide 60kg add turtle peptide, ocean fish peptide to carry out batch mixing in blender, use aluminum aluminum to pack, 10 grams every bag.
Claims (34)
1. a compositions, is characterized in that raw material comprises: turtle peptide, ocean fish peptide.
2. compositions according to claim 1, is characterized in that raw material further comprises: one or combination in any in soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator.
3. compositions according to claim 2, is characterized in that raw material comprises: turtle peptide, ocean fish peptide, soybean peptide.
4. compositions according to claim 2, is characterized in that raw material comprises: turtle peptide, ocean fish peptide, water soluble dietary fiber.
5. compositions according to claim 2, is characterized in that raw material comprises: turtle peptide, ocean fish peptide, phosphopeptide caseinate.
6. compositions according to claim 2, is characterized in that raw material comprises: turtle peptide, ocean fish peptide, soybean peptide, water soluble dietary fiber, zinc gluconate, wolfberry fruit extract, phosphopeptide caseinate, disodiumedetate, mouthfeel regulator.
7. compositions according to claim 1, is characterized in that comprising the raw material of following weight parts: turtle peptide 1-200 part, ocean fish peptide 1-100 part.
8. compositions according to claim 7, is characterized in that comprising the raw material of following weight parts: turtle peptide 10-180 part, ocean fish peptide 5-60 part.
9. compositions according to claim 8, is characterized in that comprising the raw material of following weight parts: 150 parts of turtle peptides, 20 parts of ocean fish peptide.
10. compositions according to claim 2, is characterized in that comprising the raw material of following weight parts: one or combination in any in soybean peptide 1-100 part, water soluble dietary fiber 1-100 part, zinc gluconate 0.01-0.1 part, wolfberry fruit extract 0.1-10 part, phosphopeptide caseinate 0.1-10 part, disodiumedetate 0.01-0.25 part, mouthfeel regulator 1-500 part.
11. compositions according to claim 10, is characterized in that comprising the raw material of following weight parts: one or combination in any in soybean peptide 10-70 part, water soluble dietary fiber 5-30 part, zinc gluconate 0.02-0.08 part, wolfberry fruit extract 0.5-5 part, phosphopeptide caseinate 0.2-2 part, disodiumedetate 0.05-0.22 part, mouthfeel regulator 10-200 part.
12. compositions according to claim 11, is characterized in that comprising the raw material of following weight parts: one or combination in any in 100 parts of 30 parts of soybean peptide, 15 parts of water soluble dietary fibers, 0.04 part of zinc gluconate, 2 parts of wolfberry fruit extracts, 1 part of phosphopeptide caseinate, 0.1 part of disodiumedetate, mouthfeel regulator.
13. compositionss according to claim 3, is characterized in that comprising the raw material of following weight parts: turtle peptide 1-200 part, ocean fish peptide 1-100 part, soybean peptide 1-100 part.
14. compositionss according to claim 13, is characterized in that comprising the raw material of following weight parts: turtle peptide 10-180 part, ocean fish peptide 5-60 part, soybean peptide 10-70 part.
15. compositionss according to claim 14, is characterized in that comprising the raw material of following weight parts: 150 parts of turtle peptides, 20 parts of ocean fish peptide, 30 parts of soybean peptide.
16. compositionss according to claim 4, is characterized in that comprising the raw material of following weight parts: turtle peptide 1-200 part, ocean fish peptide 1-100 part, water soluble dietary fiber 1-100 part.
17. compositionss according to claim 16, is characterized in that comprising the raw material of following weight parts: turtle peptide 10-180 part, ocean fish peptide 5-60 part, water soluble dietary fiber 5-30 part.
18. compositionss according to claim 17, is characterized in that comprising the raw material of following weight parts: 150 parts of turtle peptides, 20 parts of ocean fish peptide, 15 parts of water soluble dietary fibers.
19. compositionss according to claim 5, is characterized in that comprising the raw material of following weight parts: turtle peptide 1-200 part, ocean fish peptide 1-100 part, phosphopeptide caseinate 0.1-10 part.
20. compositionss according to claim 19, is characterized in that comprising the raw material of following weight parts: turtle peptide 10-180 part, ocean fish peptide 5-60 part, phosphopeptide caseinate 0.2-2 part.
21. compositionss according to claim 20, is characterized in that comprising the raw material of following weight parts: 150 parts of turtle peptides, 20 parts of ocean fish peptide, 1 part of phosphopeptide caseinate.
22. compositions according to claim 6, is characterized in that comprising the raw material of following weight parts: turtle peptide 1-200 part, ocean fish peptide 1-100 part, soybean peptide 1-100 part, water soluble dietary fiber 1-100 part, zinc gluconate 0.01-0.1 part, wolfberry fruit extract 0.1-10 part, phosphopeptide caseinate 0.1-10 part, disodiumedetate 0.01-0.25 part, mouthfeel regulator 1-500 part.
23. compositions according to claim 22, is characterized in that comprising the raw material of following weight parts: turtle peptide 10-180 part, ocean fish peptide 5-60 part, soybean peptide 10-70 part, water soluble dietary fiber 5-30 part, zinc gluconate 0.02-0.08 part, wolfberry fruit extract 0.5-5 part, phosphopeptide caseinate 0.2-2 part, disodiumedetate 0.05-0.22 part, mouthfeel regulator 10-200 part.
24. compositions according to claim 23, is characterized in that comprising the raw material of following weight parts: 150 parts of turtle peptides, 20 parts of ocean fish peptide, 30 parts of soybean peptide, 15 parts of water soluble dietary fibers, 0.04 part of zinc gluconate, 2 parts of wolfberry fruit extracts, 1 part of phosphopeptide caseinate, 0.1 part of disodiumedetate, 100 parts of mouthfeel regulators.
25. according to the arbitrary described compositions of claim 1-24, it is characterized in that: peptide molecular weight≤10000Dalton.
26. compositionss according to claim 25, is characterized in that: peptide molecular weight≤1000Dalton.
27. according to the arbitrary described compositions of claim 1-24, it is characterized in that: add adjuvant to make any one in oral liquid, granule, capsule, tablet, powder, dispersible tablet, syrup.
The preparation method of the oral liquid described in 28. claim 27, is characterized in that comprising the following steps:
(1) preparation: take supplementary material, standardize solution adds water;
(2) filter: with 30 μ m-0.2 μ m filter materials filtrations, fill;
(3) sterilizing: 100-125 DEG C, sterilizing 5-60 minute.
The preparation method of 29. oral liquids according to claim 28, is characterized in that: with 10 μ m-0.15 μ m filter materials filtrations; 102-121 DEG C, sterilizing 8-50 minute.
The preparation method of 30. oral liquids according to claim 29, is characterized in that: with 0.2 μ m filter material filtration; 105 DEG C, sterilizing 30 minutes.
The arbitrary described compositions of 31. claim 1-24 is in the purposes of preparing in anti-tumor health care product or medicine or product.
The arbitrary described compositions of 32. claim 1-24 strengthens the purposes in immunologic function health product or medicine or product in preparation.
The arbitrary described compositions of 33. claim 1-24 is improved the purposes in health product or medicine or the product of radiation treatment or chemotherapy untoward reaction in preparation.
34. purposes according to claim 33, wherein untoward reaction is gastrointestinal tract and hemopoietic system symptom.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210498458.5A CN103845721A (en) | 2012-11-29 | 2012-11-29 | Composition for control of radiation damage or chemotherapy damage and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210498458.5A CN103845721A (en) | 2012-11-29 | 2012-11-29 | Composition for control of radiation damage or chemotherapy damage and preparation method thereof |
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| CN103845721A true CN103845721A (en) | 2014-06-11 |
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| CN116036247A (en) * | 2023-02-03 | 2023-05-02 | 江中药业股份有限公司 | Composition for inhibiting inflammatory response, promoting angiogenesis and wound healing and application thereof |
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| CN116036247A (en) * | 2023-02-03 | 2023-05-02 | 江中药业股份有限公司 | Composition for inhibiting inflammatory response, promoting angiogenesis and wound healing and application thereof |
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