CN1557340A - Growth factor and immuneglobin preparation - Google Patents
Growth factor and immuneglobin preparation Download PDFInfo
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- CN1557340A CN1557340A CNA2004100158543A CN200410015854A CN1557340A CN 1557340 A CN1557340 A CN 1557340A CN A2004100158543 A CNA2004100158543 A CN A2004100158543A CN 200410015854 A CN200410015854 A CN 200410015854A CN 1557340 A CN1557340 A CN 1557340A
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Abstract
The present invention is the method of preparing high bioactivity growth factor and immue globulin form cow colostrum and the application of the preparation. The present invention features that cow colostrum in 72 hr after parturition is collected, and through centrifugally defatting, regulating pH value to acid, heating to solidify casein, centrifugally filtering or filtering with filtering cloth to eliminate casein, dilution, collecting supernatant, concentration with low molecular weight ultrafiltering membrane, and further processing, various preparations in different forms, including concentrated preparation, dry powder preparation, spray preparation, suppository, plaster, etc. may be prepared. The present invention may be used in the treatment of different bacterial and viral infectious diseases.
Description
Affiliated technical field
The present invention relates to the preparation method and the application thereof of a kind of somatomedin and immunoglobulin preparation, relate to the somatomedin and the method for immunoglobulin preparation and the application of said preparation that from cattle colostrums, prepare high bioactivity with a kind of especially.
The invention technical background
Colostrum is meant the preceding liquid of the excretory breast of parent in 72 hours branch puerperiums.Science and medical research for many years found, cattle colostrums contains the bioactive substance of multiple preciousness, have health care, disease-resistant, the anti-ageing effect of waiting for a long time, its effectiveness can compare favourably with human milk, and some research worker is thought the food of useful health that Lac Bovis seu Bubali is up to now to be found.
The research of cattle colostrums starts from the seventies, the various countries scientist has remarkable progress at the cattle colostrums composition and to the research aspect the human body effect over more than 30 year, comprehensive present international achievement in research, the active substance in first Ruzhong can be divided into two big classes: immune factor and somatomedin.
Immune factor in the cattle colostrums can be resisted virus, antibacterial, yeast, fungus, anaphylactogen and other toxin, just 5 kinds of immunoglobulins are contained in the Ruzhong, be respectively IgG, IgD, IgM, IgA and IgE, wherein IgG content is the highest, in human colostrum, IgG only accounts for about 2%, IgG content can reach 8%-25% in cattle colostrums, and immunoglobulin all has good defense reaction for viral infection, bacterial infection, parasite and yeast, and immunoglobulin should minimumly reach 20% in the high-quality cattle colostrums; At least the antibody that contains 19 kinds of pathogenic bacterium in the cattle colostrums comprises Salmonella, escherichia coli and streptococcic antibody.Studies show that, natural Ruzhong IgG just has than the broad-spectrum fungistatic effect, particularly cause the pathogenic microorganism of intestines and stomach disease to have good inhibition and killing effect to great majority, according to " colostrum functional food " document, IgG can suppress helicobacter pylori, jejunitis Campylobacter, yersinia enterocolitica, escherichia coli, cause Salmonella enteritidis in the cattle colostrums, can prevent and treat gastric ulcer, chronic gastritis, jejunitis, enterocolitis, alimentary toxicosis etc.Also contain rich dried meat amino acid polypeptide (PRP) in the cattle colostrums, can regulate the human thymocyte activity and the enhancing immune function of human body; The breast ferrum dawn is effective to diseases such as cancer, acquired immune deficiency syndrome (AIDS), herpes, confirmed fatigues in vain; There is the fine effect of resisting at the milk surum dawn for cancer and virus in vain; And contain immunostimulant such as cytokine and lysozyme and regulate material.
Also contain abundant somatomedin in the cattle colostrums, medical research shows that the important somatomedin in the cattle colostrums is except helping normal growth, and reparation has important effect to tissue.The abundantest somatomedin of content is quasi-insulin growthing factor I and II (GIF-I and GIF-II) in the cattle colostrums, and they help body to carry out lipid metabolism and glucose metabolism effectively, increase glucose utilization, strengthen carbohydrate tolerance.Also contain transforming growth factor (TGFA﹠amp in the cattle colostrums; B), platelet inductive factor (PDGF), epidermal growth factor, Sargramostim etc.Somatomedin is that a class is the compositions of ultimate unit with the peptide, as the term suggests it can promote the growth of various cell masses in the body, segmentation is got up, and in fact its function is diversified, and its composition and effect are similar, often complement each other.Platelet derived growth factor (PDGF) is a kind of cation of finding in the circulation granule of platelet, thermally-stabilised material, having stimulates the dawn white matter synthetic and stimulate the white effect that generates of collagen dawn by fibroblast, and it also rises fibroblast, smooth muscle cell and neurogliocyte and promotes cell division and chemotaxis.The differentiation of quasi-insulin growthing factor I and II (GIF-I, II) on cell proliferation has facilitation, wound healing there is obvious benefit, epidermal growth factor (EGF) can promote epidermal growth, and undermined skin repair is had facilitation, transforming growth factor A and B (TgFA﹠amp; B) significant to mucosa growth and tissue injury repair process.Fibroblast growth factor can promote to organize normal growth, regeneration, accelerates that old and feeble injured muscle, skin colloid are former, cartilage and nervous tissue repair.Neurotrophic growth factor (NGF) can stimulate the sympathetic ganglion growth.
Many tame foreign study achievements confirm, immunoglobulin is one group and has the simultaneously very sensitive again dawn white matter of high activity, be particularly suitable for the prevention and the treatment of antibacterial and virus disease, there has been researcher to use their treatments and antibacterial or virus-associated diseases at present, as diarrhoea, particularly relevant diarrhoea, traveler's diarrhea, baby's toxic diarrhea, harmonization of the stomach intestinal ulcer, non-ulcerative gastroenteritis, senile chronic bronchitis etc. with acquired immune deficiency syndrome (AIDS) or other immunodeficiency.
There is multinomial technology to be used for extracting bioactive substance in the preparation cattle colostrums at present, United States Patent (USP) 4,526,715 have introduced a kind of method for preparing immunoglobulin from cattle colostrums by several chromatography steps.United States Patent (USP) 3,128,230 have introduced a kind of product and method that can be used for treating various bacterial infections that prepare from cattle colostrums.DRP 2,813,284 and European patent EP-A0,046,909 and EP-A0,064,103 has narrated the method that contains the immunoglobulin product from the preparation of first Ruzhong, and this product can be used as the infection product.
As stated above, begin to prepare effective immunoglobulin product from cattle colostrums or from the process cattle colostrums of immunity inoculation, in order to reach the effect of anti-pathogenic bacterium or anti-Causative virus, must inject a strain or many strains antibacterial to animal earlier, or its antigen, so that can produce an a kind of antibody or a group-specific antibody, the activity of the product that obtains with this method is relevant with the immune kind of use, application is restricted, and product purpose is single.
In the technology that Chinese invention patent (application number 95100461) is announced, the removal cheese dawn is incomplete in vain, and uses white enzyme of stomach dawn and can damage the immunoglobulin activity.
And also be rich in multiple somatomedin and active polypeptide in the cattle colostrums, these biological active matter mass-energy help the local organization reparation, promote the dawn synthetic in vain, alleviate or recover the local organization 26S Proteasome Structure and Function, collaborative with immunoglobulin, can effectively promote the recovery from illness of disease, not have as yet in the present disclosed technology and see similar report.
In addition, present technology only is applied in gastrointestinal tract with immunoglobulin, if prepare antibacterial or viral infection that suitable dosage form also can be applied to hollow organs such as respiratory tract, urinary tract, vagina, because its natural characteristic, nontoxic pair of effect should obtain good effect.
Technical problem to be solved by this invention is, from cattle colostrums, prepare high-purity and high antibody activity immunoglobulin product by simple and economical method, preserve the wherein somatomedin and the active polypeptide of high bioactivity simultaneously, when antagonism, killing local malignant bacteria and Causative virus, the help local organization is repaired, promote the dawn synthetic in vain, fast quick-recovery local organization 26S Proteasome Structure and Function.And use this product to need not to carry out immunity inoculation in advance, and this formulation preparation can be become be fit to the multiple dosage form that a plurality of organs of human body are used, be widely used in the treatment of various bacteria and viral relevant disease.
Summary of the invention
The technical solution adopted for the present invention to solve the technical problems is, collect cattle and divide colostrum in 72 hours puerperiums, centrifugal degreasing, regulate PH to acid state with sour agent, simultaneously, it is white to add the thermocoagulation cheese dawn, remove the white back dilution of cheese dawn with centrifugation or filter cloth filter type, collect supernatant, adopt the low-molecular-weight supermicro filtration membrane to concentrate, prepare different dosage forms such as liquid concentrated formula or dry powder formulations, spray agent, drug suppository, skin patch according to the different purposes of dosage form respectively.
The technical solution adopted for the present invention to solve the technical problems also can realize by following approach, the cattle colostrums centrifugation defat of collecting, the defat cattle colostrums dilutes with distilled water, add acid supplement in the skimmed colostrum of dilution back and adjust PH, heating simultaneously, making the cheese dawn solidify back centrifugation in vain, to remove the cheese dawn white, or filter with filter cloth after the standing over night that to remove the cheese dawn white, concentrates the above-mentioned various dosage forms of back preparation with the low-molecular-weight supermicro filtration membrane after collecting supernatant.
Technical characterictic of the present invention is to collect cattle colostrums, employing centrifugation removal fat, centrifugal speed is 2000～4000 rev/mins, preferred 3000 rev/mins, centrifugation time 15～30 minutes, preferred 20 minutes, adopt frozen centrifugation equipment centrifugal under freezing state, the fat on surface is removed in centrifugal back, adds sour agent, for example 2N HCL or other acidic formulation, adjust concentrate PH to faintly acid, PH3.8～5.2, preferred 4.5～4.8, adjust the concentrate pH value after, it is white to add the heat evolution cheese dawn, 40 ℃～55 ℃ of heating-up temperatures, best 45 ℃～50 ℃, 15 minutes～90 minutes heat time heating time, best 30～45 minutes, preferably be heated to and see that whey obviously separates, then the cheese dawn of separating out with centrifuge centrifugation white, 3000～6000 rev/mins of centrifugal speeds, preferred 5000 rev/mins, collect centrifuged supernatant.
Technical characterictic of the present invention is that technical solution of the present invention also can realize that the defat cattle colostrums dilutes with 1 times～5 times distilled waters from following approach, preferred 2 times～3 times, regulate pH value to 4.6～4.9, preferred 4.7～4.8,30 ℃～40 ℃ of meanwhile heat, continue 15～45 minutes, 4 ℃ of storages of deposit in then and spend the night, with nylon filter cloth or other filter clothes, or it is white to remove the sedimentary cheese dawn with centrifugation, the supernatant molecular weight is that 4000～6000 ultrafilter membrane concentrates, and lyophilization powder process.
Technical characterictic of the present invention is that preparation of the present invention can also be prepared into injection, is used for the treatment of animal infectious disease.
Technical characterictic of the present invention is, the drying means of liquid concentrate in the last process in the above-mentioned preparation method of the present invention, both can adopt above-mentioned freeze-drying method, also can adopt spray drying process, these two kinds of drying meanss can not influence the biological activity of immunoglobulin, but spray-dired method, may influence the biological activity of somatomedin in the preparation or other active polypeptide, therefore, when preserving in the preparation somatomedin and immunoglobulin biological activity simultaneously as hope, should adopt freeze-drying method, reduce cost if wish, should select spray drying process when need not to preserve somatomedin class polypeptide active again, temperature can not surpass 160 °F during spray drying, in order to avoid infringement immunoglobulin activity.
Technical characterictic of the present invention is, adopt the preparation that is rich in somatomedin and immunoglobulin of method for preparing, also can directly use with the form of liquid concentrate, for example be prepared into oral concentrated solution mode and be applied to treat gastroenteropathy, be prepared into the treatment that the dropping liquid form is used for the infectious disease of eyes, oral cavity, ear.Also can be prepared into suppository and be used for the intestinal infection treatment of diseases.Be prepared into the membranous type agent and be used for skin injury and skin infection treatment of diseases.Be prepared into the treatment that spray form is used for respiratory tract infectious disease, also can add in the storage medicine device of spray atomization, the mode that adopts ullrasonic spraying to atomize is treated the infectious disease of respiratory tract, oral cavity, throat.The preparation of above-mentioned preparation adopts those skilled in the art to understand, and ripe dosage form manufacturing technology on top of, adds the adjuvant of various medical-grade, uses the equipment preparation of medical-grade.
Technical characterictic of the present invention is, that adopts method for preparing is rich in somatomedin and immunoglobulin preparation, comprise that liquid condition concentrate or dry powder all can be used for the treatment of multiple gastroenteropathy, multiple and antibacterial or the relevant gastroenteropathys of virus such as these gastroenteropathys include but not limited to suffer from diarrhoea, alimentary toxicosis, infantile diarrhea, traveler's diarrhea, the diarrhoea of being correlated with, gastrointestinal ulceration, chronic gastritis, jejunitis, enterocolitis, yersinia infection with acquired immune deficiency syndrome (AIDS) or other immune deficiency disorders.
The invention has the beneficial effects as follows that compared with prior art, the preparation of the present invention's preparation not only contains a large amount of immunoglobulins, but also contains the raised growth factor, when killing local antibacterial and virus, can help to repair local organization.
Specific embodiments
Embodiment 1
Collect cattle colostrums, centrifugal degreasing, transfer PH to 4.7 with 2N HCL, heat 40 ℃ simultaneously and continue 30 minutes, with centrifugal 20 minutes of 5000 rev/mins of speed, the collection supernatant is used, 0.9% sodium chloride solution is diluted to the twice of original volume, it with molecular weight 5000 supermicro filtration membrane, on the staggered tangent plane fluid ultrafiltration apparatus that U.S. Pall company produces, be concentrated into original volume 10%, send into freeze drying equipment and prepare white powder, include abundant, the somatomedin of high bioactivity and immunoglobulin, adopt para-insulin factor radioimmunity to detect box and measure, contain 0.2 microgram insulin like growth factor in every gram dry powder.
Embodiment 2
Collect cattle colostrums, centrifugal degreasing, with double amount distilled water dilution, transfer PH to 4.8 with 2N HCL, obtain supernatant, in 38 ℃ of heating 40 minutes, leave standstill to store in 4 ℃ then and spend the night, supernatant is removed white coarse granule of sedimentary cheese dawn by the nylon cloth filter, is concentrated into original volume 30% with the supermicro filtration membrane of molecular weight 6000 in the plate and frame ultrafiltration apparatus, uses the spray drying method spray drying, inlet temperature is 160 °F during spray drying, 130 of outlet temperatures, result obtain containing the immunoglobulin dry powder of 80% purity, before the spray drying after the biological activity of preparation and the spray drying biological activity of preparation consistent.
Embodiment 3
Collect and divide 10 hours puerperiums cattle colostrums, with 3000 rev/mins of speed centrifugalize fat, regulate PH to 4.7 (Ruzhong adds about 54m11N HCL at the beginning of every liter) with 1N HCL, heat 42 ℃ simultaneously and continue 35 minutes, white with the cheese dawn that 5000 rev/mins of speed centrifugations are solidified, collect supernatant, normal saline dilution with 2 times of amounts, supernatant after the dilution is just filtered with hollow fiber column, doughnut average pore size 0.4 μ m, surface area 3m
2, fibre diameter 1.2mm, working pressure surpass atmosphere 0.2-0.6 crust.Collect filtrate, carry out the second time by the system that is made up of three hollow fiber columns again with the normal saline dilution of 6 times of amounts and filter, the separation extreme of each post is 60, and 000D, surface area are 1.4m
2, be concentrated to and filter 2 times of preceding initial capacities for the first time.Through measuring, dawn white matter content is 6% in the resulting concentrated solution, carries out filter-sterilized with known method, or adopts pasteurization.
Embodiment 4:
Prepare concentrated solution by embodiment 3 methods, add in the medical grade heating polyglycol solution, mix and stir, pour into and preset suppository mould, room temperature cooling back preparation suppository can be applied to all kinds of infection of woman uterus or vagina.
Embodiment 5
Prepare concentrated solution by embodiment 3 methods, mix stirring with the medical grade aerosol solution, pressurization is filled in the gas cylinder, and the preparation aerosol can be used for oral cavity, throat, upper respiratory tract infection treatment of diseases, can also be used to anti-curing cold and upper respiratory tract infection.
Embodiment 6
Press the dry powder of embodiment 1 method preparation, preparation 200m15% dawn white matter sterile solution, this solution are stirred by 15g dry powder adding 100ml distilled water and are made into, it is composed as follows: total dawn white matter 4.2g/100ml, IgG 3.0g/100ml, IgA0.35g/100ml, IgM 0.96g/100ml.
Embodiment 7
The staphylococcus aureus supernatant that application contains toxin carries out toxin neutralization test, adopt the preparation of embodiment 1 preparation to compare with people's immunoglobulin preparation, for the required people's immunoglobulin consumption of 50% haemolysis (toxin neutralization), be three times of preparation consumption of the present invention.
Embodiment 8
Carry out toxin neutralization test with embodiment 1 preparation and the toxin supernatant that contains gram negative bacteria (pseudomonas aeruginosa), the immunoglobulin of the inventive method preparation can produce 100% toxin neutralization at low concentration very.
Claims (8)
1. the present invention relates to a kind of from cattle colostrums, the prepare method of high bioactivity somatomedin and immunoglobulin and the application of said preparation, it is characterized in that, collect cattle and divide colostrum in 72 hours puerperiums, adopt centrifugal degreasing, regulate PH to acid state with sour agent, simultaneously, it is white to add the thermocoagulation cheese dawn, remove the white back dilution of cheese dawn with centrifugation or filter cloth filter type, collect supernatant, adopt the low-molecular-weight supermicro filtration membrane to concentrate, prepare liquid concentrated formula or dry powder formulations, spray agent respectively according to the different purposes of dosage form, drug suppository, different dosage forms such as skin patch.
2. in accordance with the method for claim 1, it is characterized in that, the technical solution adopted for the present invention to solve the technical problems also can realize by following approach, the cattle colostrums centrifugation defat of collecting, the defat cattle colostrums dilutes with distilled water, add acid supplement in the skimmed colostrum of dilution back and adjust PH, heating simultaneously, it is white that the cheese dawn was removed in centrifugation after the cheese dawn was solidified in vain, or it is white filter to remove the cheese dawn with filter cloth after the standing over night, concentrates the above-mentioned various dosage forms of back preparation with the low-molecular-weight supermicro filtration membrane after collecting supernatant.
3. in accordance with the method for claim 1, it is characterized in that, collect cattle colostrums, adopt centrifugation to remove fat, centrifugal speed is 2000～4000 rev/mins, preferred 3000 rev/mins, centrifugation time 15～30 minutes, preferred 20 minutes, adopt frozen centrifugation equipment centrifugal under freezing state, the fat on surface is removed in centrifugal back, add sour agent, for example 2N HCL or other acidic formulation are adjusted concentrate PH to faintly acid, PH3.8～5.2, preferred 4.5～4.8, after adjusting the concentrate pH value, it is white to add the heat evolution cheese dawn, 40 ℃～55 ℃ of heating-up temperatures, best 45 ℃～50 ℃, 15 minutes～90 minutes heat time heating time, best 30～45 minutes, preferably be heated to and see that whey obviously separates, the cheese dawn of separating out with centrifuge centrifugation is white then, 3000～6000 rev/mins of centrifugal speeds, are collected centrifuged supernatant by preferred 5000 rev/mins.
4. in accordance with the method for claim 1, it is characterized in that, technical solution of the present invention also can realize from following approach, the defat cattle colostrums dilutes with 1 times～5 times distilled waters, preferred 2 times～3 times, regulate pH value to 4.6～4.9, preferred 4.7～4.8,30 ℃～40 ℃ of meanwhile heat, continue 15～45 minutes, 4 ℃ of storages of deposit in then and spend the night, with nylon filter cloth or other filter clothes, or it is white to remove the sedimentary cheese dawn with centrifugation, and the supernatant molecular weight is that 4000～6000 ultrafilter membrane concentrates, and lyophilization powder process.
5. in accordance with the method for claim 1, it is characterized in that, the drying means of liquid concentrate in the last process in the above-mentioned preparation method of the present invention, both can adopt above-mentioned freeze-drying method, also can adopt spray drying process, these two kinds of drying meanss can not influence the biological activity of immunoglobulin, but spray-dired method, may influence the biological activity of somatomedin in the preparation or other active polypeptide, therefore, when preserving in the preparation somatomedin and immunoglobulin biological activity simultaneously, should adopt freeze-drying method as hope, if wish to reduce cost, should select spray drying process when need not to preserve somatomedin class polypeptide active again.
6. in accordance with the method for claim 1, it is characterized in that, adopt the preparation that is rich in somatomedin and immunoglobulin of method for preparing, also can directly use with the form of liquid concentrate, for example be prepared into oral concentrated solution mode and be applied to treat gastroenteropathy, be prepared into the treatment that the dropping liquid form is used for the infectious disease of eyes, oral cavity, ear.Also can be prepared into suppository and be used for the intestinal infection treatment of diseases.Be prepared into the membranous type agent and be used for skin injury and skin infection treatment of diseases.Be prepared into the treatment that spray form is used for respiratory tract infectious disease, also can add in the storage medicine device of spray atomization, the mode that adopts ullrasonic spraying to atomize is treated the infectious disease of respiratory tract, oral cavity, throat.The preparation of above-mentioned preparation adopts those skilled in the art to understand, and ripe dosage form manufacturing technology on top of, adds the adjuvant of various medical-grade, uses the equipment preparation of medical-grade.
7. in accordance with the method for claim 1, it is characterized in that, that adopts method for preparing is rich in somatomedin and immunoglobulin preparation, comprise that liquid condition concentrate or dry powder all can be used for the treatment of multiple gastroenteropathy, multiple and antibacterial or the relevant gastroenteropathys of virus such as these gastroenteropathys include but not limited to suffer from diarrhoea, alimentary toxicosis, infantile diarrhea, traveler's diarrhea, the diarrhoea of being correlated with, gastrointestinal ulceration, chronic gastritis, jejunitis, enterocolitis, yersinia infection with acquired immune deficiency syndrome (AIDS) or other immune deficiency disorders.
In accordance with the method for claim 1 in, preparation of the present invention can also be prepared into injection, is used for the treatment of animal infectious disease.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2004100158543A CN1557340A (en) | 2004-01-15 | 2004-01-15 | Growth factor and immuneglobin preparation |
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| Application Number | Priority Date | Filing Date | Title |
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| CNA2004100158543A CN1557340A (en) | 2004-01-15 | 2004-01-15 | Growth factor and immuneglobin preparation |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8252341B2 (en) | 2004-09-14 | 2012-08-28 | Paul Brazeau | Isolation of growth and differentiating factors from colostrum |
| CN103338778A (en) * | 2010-12-09 | 2013-10-02 | 赞邦股份公司 | Multipurpose gel for vaginal dryness with direct and delayed effect |
| CN111067839A (en) * | 2020-01-16 | 2020-04-28 | 广东丽姿生物科技有限公司 | Conditioning repairing agent and preparation method thereof |
-
2004
- 2004-01-15 CN CNA2004100158543A patent/CN1557340A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8252341B2 (en) | 2004-09-14 | 2012-08-28 | Paul Brazeau | Isolation of growth and differentiating factors from colostrum |
| CN103338778A (en) * | 2010-12-09 | 2013-10-02 | 赞邦股份公司 | Multipurpose gel for vaginal dryness with direct and delayed effect |
| CN111067839A (en) * | 2020-01-16 | 2020-04-28 | 广东丽姿生物科技有限公司 | Conditioning repairing agent and preparation method thereof |
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