WO2012024817A1 - 全蛋蛋白肽及其制备方法和应用 - Google Patents
全蛋蛋白肽及其制备方法和应用 Download PDFInfo
- Publication number
- WO2012024817A1 WO2012024817A1 PCT/CN2010/001287 CN2010001287W WO2012024817A1 WO 2012024817 A1 WO2012024817 A1 WO 2012024817A1 CN 2010001287 W CN2010001287 W CN 2010001287W WO 2012024817 A1 WO2012024817 A1 WO 2012024817A1
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- WIPO (PCT)
- Prior art keywords
- whole egg
- egg protein
- product
- peptide
- protein peptide
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/02—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from eggs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/57—Birds; Materials from birds, e.g. eggs, feathers, egg white, egg yolk or endothelium corneum gigeriae galli
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/012—Hydrolysed proteins; Derivatives thereof from animals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/465—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from birds
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
Definitions
- the invention relates to a preparation method and application of a novel peptide immunological whole egg protein peptide.
- peptide immunostimulants belong to polypeptide hormones, mainly drugs such as thymosin, thymopentin, immunoglobulin, gamma globulin, human serum albumin, interferon, tumor necrosis factor, interleukin I, and leukocyte mediator. Prime II, interleukin III, etc.
- drugs such as thymosin, thymopentin, immunoglobulin, gamma globulin, human serum albumin, interferon, tumor necrosis factor, interleukin I, and leukocyte mediator.
- These peptide immunizing agents are isolated from the gland, tissue or blood of an animal. Most of these peptide drugs are injections, which can only be used in certain patients. During use, they are prone to rejection, allergies and side effects. They must be used with caution and must be used under strict supervision by a doctor.
- the object of the present invention is to provide a peptide-based immunizing agent whole egg protein peptide and a preparation method thereof.
- the whole egg protein peptide provided by the present invention is prepared according to the following method: Enzymatic degradation of whole egg protein powder by a composite plant protease to obtain the whole egg protein peptide.
- the composite plant protease may be composed of papain, ficin and pineapple protease, and the enzyme activity ratio thereof is (100 to 1.2 million) U: (30 to 400,000) U: (560 to 7 million) U.
- the whole egg protein powder can be prepared from at least one of the following bird eggs: eggs, duck eggs, goose eggs, quail eggs, sparrow eggs, pigeon eggs, spotted eggs and ostrich eggs.
- the mass fraction of whole egg protein powder to water is 1: 8-10, and the amount of each enzyme required to digest the whole egg protein powder is: papain 5 ⁇ 6 Million U, fig protease 1. 5 ⁇ 2 million U, bromelain 28 ⁇ 3 million U. 5 ⁇
- the enzyme is catalyzed by a temperature of 48 to 50 ° C, a time of 3 to 4 hours, a pH of 7. 5 ⁇ 8.
- the enzymatic hydrolysate is also subjected to sterilization and enzyme inactivation treatment, and then the enzymatic hydrolysate is cooled and frozen to obtain the final whole egg protein peptide.
- the freezing temperature was 4 to 8 ° C for 48 hours; the sterilization and enzyme inactivation conditions were: temperature 100 ° C, time 10 minutes.
- the enzyme activity unit U refers to the amount of enzyme required to convert 1 micromolar substrate per minute or to convert 1 micromolar of the relevant group under specific conditions (25 ° C, pH 7.0).
- Enzyme The whole egg protein peptide obtained by the invention contains a polypeptide, an oligopeptide, 20 kinds of amino acids (8 of which are essential amino acids which cannot be synthesized), and a plurality of vitamins (VA, VC, VE, Bl, B2, B6, B12), calcium and a variety of organic and natural trace elements (CPPS, zinc, selenium, magnesium, copper, iron, manganese, etc., which are naturally chelated with proteins).
- vitamins VA, VC, VE, Bl, B2, B6, B12
- CPPS organic and natural trace elements
- the polypeptides and oligopeptides have a relative molecular mass of less than 100 ODA and consist of 2-6 amino acids.
- the whole egg protein peptide may be present in the form of a concentrate, a lyophilized powder, a spray dried dry powder, a liquid or a solid.
- Another object of the invention is to provide the use of the whole egg protein peptide.
- the use of the whole egg protein peptide provided by the present invention is its use in the preparation of products for enhancing immunity.
- the product for enhancing immunity provided by the present invention the active ingredient thereof comprises the whole egg protein peptide provided by the present invention.
- the product can be a pharmaceutical or health supplement.
- the mass ratio of whole egg protein peptide, jujube polysaccharide and anthraquinone polysaccharide may be (900-1200): (8-30): (3-15).
- An immunopotentiating drug or nutraceutical prepared by using whole egg protein peptide as an active ingredient, and one or more pharmaceutically acceptable carriers may be added to the above-mentioned drug or health care product as needed.
- the carrier includes conventional diluents, excipients, fillers, binders, humectants, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants and the like in the pharmaceutical field.
- the immunopotentiating drug can be formulated into various forms such as an oral solution, a tablet, a granule, a capsule, a paste, a film, and the like.
- the above various dosage forms of the drug can be prepared according to a conventional method in the pharmaceutical field, and the obtained drug has no bitter taste.
- Enzymatic hydrolysis using whole egg protein powder as a raw material is a remarkable feature of the present invention.
- the protein scores of the above eggs are close to 100 points.
- the protein is close to human protein and is easily absorbed and utilized by the body.
- the choice of whole eggs is a scientific choice. If only egg white protein is selected, the protein and many nutrients in the egg yolk will be lost. If only egg yolk is selected, the protein in the egg white is also lost. Choosing whole eggs means choosing a complete nutrition system. Not only is nutrition comprehensive, but each nutrient can exert synergy with each other.
- the invention adopts compound plant proteases, namely papain, ficin and pineapple eggs.
- the white enzyme, the three enzyme scientific formula forms a composite enzyme preparation, and the whole egg protein powder is catalytically degraded to obtain a whole egg protein peptide.
- the whole egg protein peptide can replace the polypeptide hormone immunosuppressive agent and function as an immunomodulatory agent for the polypeptide hormone immunosuppressive agent.
- the polypeptide hormone immunosuppressive agent is a medicine and can only be used for a patient, and the product of the invention can be made into an oral polypeptide nutritional immunizing agent, which not only can treat the patient, but also can enhance the immunity to the disease-free person. The role of force, disease prevention and health.
- This product is an oral preparation that is easier to carry and safe to use than the injection. It does not cause rejection, allergies or other side effects during use, and does not need to be used under the supervision of a doctor. It can be used for many diseases and sub-health states caused by low immune function, immune dysfunction, decreased immune function, impaired immune organs, etc. It can also be used for protein-deficient people, people with many diseases caused by protein deficiency and sub-health status.
- It can be used to prevent colds, can be used after exercise, postpartum, post-ill, post-operative population, promote "negative nitrogen balance", postpartum recovery, post-mortem recovery, wound healing, liver protection, hangover, appetite recovery, sleep promotion It has a good effect on restoring kidney function, stimulating the phagocytic ability of macrophages, inhibiting the growth of tumor cells, and raising white blood cells.
- Figure 1 is a flow chart showing the production process of the whole egg protein peptide of the present invention.
- Example 1 Preparation of whole egg protein peptide oral liquid for enhancing immunity
- the whole egg protein powder (freeze-dried dry powder or spray-dried dry powder) into the fermenter, add water equivalent to 10 times the mass of the whole egg protein powder, and gradually increase the temperature to 5 CTC, and then enzymatically digest the whole egg protein per gram.
- the amount of various enzymes required for the powder is: 50,000 U of papain, 20,000 U of fig protease, and 350,000 U of Bromelain.
- the complex protease is added to the fermenter for enzymatic hydrolysis. The temperature is always maintained at 50 V. After 4 hours of enzymatic hydrolysis, it is sterilized and inactivated by enzyme, then cooled to 60 ° C for barreling, then cooled to room temperature and placed in a freezer.
- Example 2 Preparation of whole egg protein peptide oral liquid for enhancing immunity containing jujube polysaccharide and lycium polysaccharide
- Example 3 Preparation of an immune-enhancing whole egg protein peptide oral solution containing jujube polysaccharide and lycium polysaccharide
- the formulated functional ingredients are composed of: 1000 mg ⁇ 10% of whole egg protein peptide per 100 ml of oral liquid, 15 mg of jujube polysaccharide, 10 mg of medlar polysaccharide, and after filling and sterilizing at 10 CTC for 10 minutes, it is enhanced.
- Immune oral solution 1000 mg ⁇ 10% of whole egg protein peptide per 100 ml of oral liquid, 15 mg of jujube polysaccharide, 10 mg of medlar polysaccharide, and after filling and sterilizing at 10 CTC for 10 minutes, it is enhanced.
- Immune oral solution 1000 mg ⁇ 10% of whole egg protein peptide per 100 ml of oral liquid, 15 mg of jujube polysaccharide, 10 mg of medlar polysaccharide, and after filling and sterilizing at 10 CTC for 10 minutes, it is enhanced.
- Immune oral solution 1000 mg ⁇ 10% of whole egg protein peptide per 100 ml of oral liquid, 15 mg of jujube poly
- Test sample Oral solution prepared in Example 1
- the organic nitrogen (%) represents the peptide and amino acid content
- the determination Refer to GB5009 for the law.
- mice Kunming mice 18-20g, male and female, a total of 20; animals were provided by the Hubei Medical Laboratory Animal Center.
- Test drug The oral solution for enhancing immunity prepared in Example 1.
- test animals were orally administered with 0.3 ml/10 g.b.w. Fasting for 12 hours before gavage, observe for one week.
- test article is a non-toxic substance.
- Test method The test drug was administered twice by intragastric administration. After the second gavage for 6 hours, the animals were sacrificed. The thoracic bone marrow was diluted with calf serum and stained with Gimsa. 1000 polychromatic red blood cells were observed under microscope. The number of micronucleated cell formation was recorded and expressed in parts per thousand. The results are shown in Table 4. Among them, the CP group was a cyclophosphamide positive control group.
- mice Kunming mice 23-25g.
- Test method After 5 days of continuous gavage, continue feeding for 30 days. The animals were sacrificed and the lateral testicular slices were taken. Each animal counted 1000 structurally intact sperm and calculated the incidence of sperm abnormalities. The results are shown in Table 5. Among them, the CP group was a cyclophosphamide positive control group.
- PCB Polychlorinated biphenyl
- the -S 9 positive control group was 3-nitro 9-fluorenone (2.4.7-TNFone), and the +S 9 positive control group was 2-aminopurine (2-AF).
- each experimental group was returned. The number of colonies did not exceed twice the number of spontaneous reversions; and there was no dose-response relationship, and the test result was negative.
- Example 6 Detection of immunomodulatory effects of whole egg protein peptide
- Test animals Kunming mice 18-22g, male and female. (provided by Hubei Medical Laboratory Animal Center)
- the dose grouping is: control group, digestive distilled water; the concentration of "oral solution prepared in 1" is 1% (ie, lOOOO/lOOml), and the low, medium and high dose groups are respectively lOOmg/kg. b. w, 300mg/kg. b. w, 900mg/kg. b. w mouth sputum sputum; the low, medium and high dose groups are equivalent to 6, 18, 54 times the recommended intake.
- the stomach was administered for one month, and then various experiments were performed.
- RESULTS MTT assay (ConA-induced spleen lymphocyte transformation test in mice)
- One-way ANOVA was performed on the mean values of absorbance differences between ConA wells and ConA wells.
- ConA induces spleen lymphocyte proliferation
- DNFB induces DTH group
- DNFB Dinitrofluorobenzene induces delayed type hypersensitivity
- Ear swelling method After sensitizing the mice with DNFB, the right ear was attacked with DNFB on the fifth day. After 24 hours, the animals were sacrificed and the left and right ear shells were cut out. The ear pieces with a diameter of 8 mm were removed with a puncher, and weighed, and the degree of DTH was expressed by the difference between the weights of the left and right ears.
- Blood coagulation method Blood coagulation method.
- the anti-volume number was calculated according to the level of serum coagulation, and the mean value of each group of antibody was analyzed by one-way ANOVA.
- Method Semi-in vivo method. Prepare 20% chicken red blood cell suspension; each mouse was intraperitoneally injected with 1 mL of the suspension. After 30 minutes, the animals were sacrificed, laparotomy, 2 mL of normal saline was intraperitoneally injected, and the average was divided into 2 slides, and incubated at 37 °C for 30 min. The cells were rinsed with physiological saline, air-dried, fixed in 1:1 acetone in methanol, stained with 4% Giemsa-phosphate buffer for 3 min, and rinsed with distilled water to dry. 100 macrophages were counted under an oil microscope, and the phagocytosis rate and phagocytic index were calculated as follows:
- Phagocytosis rate ( %) number of macrophages that phagocytose chicken red blood cells
- the phagocytic index of each group of mice was calculated according to the carbon concentration in the blood after the injection of ink for 2 min and lOmin.
- Control group 5 6.609 ⁇ 0.755
- the whole oral liquid of egg protein peptide 1) can enhance the proliferation ability of mouse spleen lymphocytes induced by ConA; 2) enhance the induction of dinitrofluorobenzene Delayed hair allergy in mice; 3) Increase serum hemolysin content in mice; 4) Enhance the ability of mouse peritoneal macrophages to engulf chicken red blood cell function and carbon clearance.
- the whole egg protein peptide prepared by the present invention can be considered to have an immunomodulatory effect.
- the whole egg protein powder is used as a substrate, and a composite plant protease: papain, ficin and bromelain is used as a substrate to form a composite enzyme preparation, and the whole egg protein powder is catalyzed and degraded to obtain a whole egg protein peptide.
- This product has high biological activity and is a A new type of immunomodulator and immunopotentiator.
- the whole egg protein peptide of the present invention can be used for the development of immunopharmaceuticals, health foods, functional foods, anti-aging and infant foods, and nutritional supplements for seriously ill patients.
- the invention can be made into a lyophilized powder by freeze-drying, thereby being made into a capsule or a tablet, and can deeply develop a tumor adjuvant therapeutic drug (enhancing immune function, enhancing radiation resistance of a patient undergoing radiotherapy and chemotherapy, increasing red and white blood cells), and anti-inflammatory drugs.
- a tumor adjuvant therapeutic drug enhancing immune function, enhancing radiation resistance of a patient undergoing radiotherapy and chemotherapy, increasing red and white blood cells
- anti-inflammatory drugs enhancing immune function, enhancing radiation resistance of a patient undergoing radiotherapy and chemotherapy, increasing red and white blood cells
- hepatitis drugs drugs for the prevention and treatment of atypical pneumonia, can also be used as other raw materials or as food additives. No matter what kind of drugs, health products, and functional foods are developed, they will have huge economic and social benefits, and their prospects are very broad.
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- Biochemistry (AREA)
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- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
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- Coloring Foods And Improving Nutritive Qualities (AREA)
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Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201080068668.XA CN103108957B (zh) | 2010-08-24 | 2010-08-24 | 全蛋蛋白肽及其制备方法和应用 |
JP2013525103A JP5927191B2 (ja) | 2010-08-24 | 2010-08-24 | 全卵タンパク質ペプチド、その調製方法及び使用 |
PCT/CN2010/001287 WO2012024817A1 (zh) | 2010-08-24 | 2010-08-24 | 全蛋蛋白肽及其制备方法和应用 |
US13/818,642 US9657086B2 (en) | 2010-08-24 | 2010-08-24 | Whole egg protein peptides, preparation method and use thereof |
EP10856270.3A EP2610349A4 (en) | 2010-08-24 | 2010-08-24 | FULL-PIPES, PREPARATION AND USE |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/CN2010/001287 WO2012024817A1 (zh) | 2010-08-24 | 2010-08-24 | 全蛋蛋白肽及其制备方法和应用 |
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WO2012024817A1 true WO2012024817A1 (zh) | 2012-03-01 |
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ID=45722802
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PCT/CN2010/001287 WO2012024817A1 (zh) | 2010-08-24 | 2010-08-24 | 全蛋蛋白肽及其制备方法和应用 |
Country Status (5)
Country | Link |
---|---|
US (1) | US9657086B2 (zh) |
EP (1) | EP2610349A4 (zh) |
JP (1) | JP5927191B2 (zh) |
CN (1) | CN103108957B (zh) |
WO (1) | WO2012024817A1 (zh) |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107164448A (zh) * | 2017-07-12 | 2017-09-15 | 邹远东 | 一种铁棍怀山药多肽及其制备方法 |
CN108244031B (zh) * | 2017-12-15 | 2020-08-28 | 中国水产科学研究院东海水产研究所 | 一种提高免疫低下模型小鼠非特异性免疫力的方法 |
CN109744492A (zh) * | 2019-01-10 | 2019-05-14 | 蕴彤本草(北京)生物科技有限公司 | 高生物活性全蛋体发酵物的生产方法及其产品与应用 |
CN109673809A (zh) * | 2019-01-10 | 2019-04-26 | 蕴彤本草(北京)生物科技有限公司 | 活血调经化淤祛斑的花优素复合多肽产品及其制备方法 |
CN113995071A (zh) * | 2021-10-29 | 2022-02-01 | 杭州倍欣生物科技有限公司 | 一种白蛋白肽饮品和制备方法以及在提升血浆白蛋白指标上的应用 |
CN118685483A (zh) * | 2024-08-29 | 2024-09-24 | 江中药业股份有限公司 | 一种具有补血功能的乌鸡肽的制备方法及应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1483830A (zh) * | 2003-07-18 | 2004-03-24 | 上海复旦新杨生物科技有限责任公司 | 一种酶法水解鸡蛋蛋清蛋白制备蛋清肽的方法 |
CN101675938A (zh) * | 2008-12-24 | 2010-03-24 | 邹远东 | 一种新型肽类免疫剂全蛋蛋白肽的制备方法和用途 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB9403935D0 (en) * | 1994-03-01 | 1994-04-20 | Sandoz Nutrition Ltd | Improvements in or relating to organic compounds |
KR101331777B1 (ko) * | 2004-12-23 | 2013-11-21 | 캄피나 네덜란드 홀딩 베.붸. | Dpp-iv를 억제하는 펩타이드가 풍부한 단백질가수분해물 및 이의 용도 |
EP1982604A1 (de) * | 2007-04-20 | 2008-10-22 | Naturheilzentrum Allgäu | Nahrungsergänzungsmittel zum Ausgleich von Nährstoffmangel |
-
2010
- 2010-08-24 CN CN201080068668.XA patent/CN103108957B/zh active Active
- 2010-08-24 WO PCT/CN2010/001287 patent/WO2012024817A1/zh active Application Filing
- 2010-08-24 JP JP2013525103A patent/JP5927191B2/ja active Active
- 2010-08-24 US US13/818,642 patent/US9657086B2/en active Active
- 2010-08-24 EP EP10856270.3A patent/EP2610349A4/en not_active Withdrawn
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1483830A (zh) * | 2003-07-18 | 2004-03-24 | 上海复旦新杨生物科技有限责任公司 | 一种酶法水解鸡蛋蛋清蛋白制备蛋清肽的方法 |
CN101675938A (zh) * | 2008-12-24 | 2010-03-24 | 邹远东 | 一种新型肽类免疫剂全蛋蛋白肽的制备方法和用途 |
Non-Patent Citations (3)
Title |
---|
MINE, Y. ET AL.: "New insights in biologically active proteins and peptides derived from hen egg", WORLD'S POULTRY SCIENCE JOURNAL, vol. 62, no. 1, March 2006 (2006-03-01), pages 87 - 95, XP008167793 * |
See also references of EP2610349A4 * |
TIAN, G. ET AL.: "Effects of mixtures of small peptides from enzymic hydrolysates of egg white immune functions of mice", CHINESE JOURNAL OF ANIMAL SCIENCE, vol. 41, no. 5, 2005, pages 14 - 17, XP008167828 * |
Also Published As
Publication number | Publication date |
---|---|
EP2610349A1 (en) | 2013-07-03 |
CN103108957A (zh) | 2013-05-15 |
CN103108957B (zh) | 2015-07-01 |
US20130209497A1 (en) | 2013-08-15 |
EP2610349A4 (en) | 2015-12-30 |
US9657086B2 (en) | 2017-05-23 |
JP5927191B2 (ja) | 2016-06-01 |
JP2013535976A (ja) | 2013-09-19 |
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