CN1442151A

CN1442151A - Mackefulan immune liquid

Info

Publication number: CN1442151A
Application number: CN03118887A
Authority: CN
Inventors: 张素琴
Original assignee: Individual
Current assignee: Individual
Priority date: 2003-04-04
Filing date: 2003-04-04
Publication date: 2003-09-17
Anticipated expiration: 2023-04-04
Also published as: CN1296059C

Abstract

An immunizing liquid "Maikefulai" used to prpare medicines in the form of injection or capsule for treating dermatopathy, improving immunity and preventing and resisting cancer is prepared from the excellent Mycobaterium phlei through culturing in the culture medium prepared from cane sugar, corn milk, diammonium hydrogen phosphate, bipotassium hydrogen phosphate, yeast and water, fermenting in the culture meium prepared from cane sugar, pepton, diammonium hydrogen phosphate, bipotassium hydrogen phosphate and water, homogenizing, and sterilizing, and its pH value is 7.0.

Description

The Mackefulan immune liquid

Technical field

The present invention relates to biomedical engineering field, particularly a kind of Mackefulan immune liquid that improves body immunity.Mackefulan is by Latin Mycobaterium phlei translation, and it is the transliteration of mycobacterium graminis formal name used at school.The microorganism that this patent is used (strain) sq-1, be mycobacterium graminis (Mycobaterium phlei), by China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) preservation, preservation date is on April 1st, 2003, and preserving number is CGMCC NO.0916.

Background technology

As everyone knows, the immunity of cell plays an important role in many infectious disease therapeutic processes, and in some subhealth states and the individuality of having fallen ill, because immunologic hypofunction makes pure chemoprophylaxis and therapeutic effect not good.20 end of the centurys, it is found that mycobacterium muramyldipeptide (MDP) has very strong immunization therapy effect, grass divides pole polysaccharide MPS (the arabinose galactose gathers polysaccharide) aspect immunomodulating, promotion hemopoietic system, supression tumor and the virus replication very big effect being arranged, and can modify damaged cell.In recent years, to the demethylation cytosine and be that the strong adjuvant effect of the oligodeoxynucleotide sequence C pG of core causes widely and pays close attention to the guanine.Experimental results show that through immunology etc.: the fermented product of mycobacterium graminis bacterial strain can significantly improve the conversion of white mice splenocyte, strengthen quantity and the function of mouse macrophage and NK, promote the secretion of tumor necrosis factor (TNF) and interleukin II (IL-2); Obviously the carbon that strengthens dinitro inductive mice delayed allergy of fluorine phenol and mice is cleaned up ability, significantly improves the content of hemolysin in the serum.Therefore, Chinese scholars does one's utmost to pay close attention to and admit the notion of immunization therapy, thereby impels BCG-polysaccharide, and nucleic acid, mycobacterium vaccine inoculation liquid are used for the upsurge of the immunization therapy of difficult and complicated illness such as bladder cancer, asthma, tuberculosis.Under the inspiration of traditional Chinese medical science Modernization Theory, ecosystem cure is also quietly given birth to.

Although Chinese patent " SHENGTAI PINGHENG YINGYANG JINGHUOSU and production method thereof " (number of patent application 98113617.6, publication number CN1207416A), making good try aspect the physiological mechanism effect of regulating human body and the raising body immunity, but, because the technology that is adopted is immature, do not contain or contain the mycobacterium graminis active ingredient of minute quantity in its product, so do not reach described goal of the invention of this patent documentation and effect.

Summary of the invention

Technical problem to be solved by this invention is: with the microbial strains mycobacterium graminis, by the consummate in addition food technology technology of fermentation engineering, genetic engineering, make a kind of carotene, flavone and V that contains needed by human body _B, V _C, V _EDeng multivitamin, and each seed amino acid, the trace element of needed by human, especially also contain the Mackefulan immune liquid of multiple components such as muramyldipeptide polysaccharide with specifc immunity function.This immune liquid technology is simple, and its product can play the physiological mechanism effect of regulating human body effectively, to improve body immunity, reaches the purpose of diseases such as antitumor, antiviral.

The technical solution adopted for the present invention to solve the technical problems is as follows:

Its preparation process comprises: adopt genetic engineering method, and microorganism (strain) sq-1 that selection-breeding one strain is good, promptly preserving number is the mycobacterium graminis (Mycobaterium phlei) of CGMCC NO.0916, is placed in the seed tank and cultivates; In fermentation tank, cultivate and ferment, obtain to contain the biological product of carotene, flavone multivitamin and muramyldipeptide polysaccharide; By sparing matter and sterilization, operation, make Mackefulan immune liquid product again.Wherein, in seed tank, the culture medium prescription of mycobacterium graminis is (weight %): sucrose 1.5～2.8, and Semen Maydis pulp 0.4～0.8, diammonium phosphate 0.15～0.4, dipotassium hydrogen phosphate 0.05～0.1, yeast extract 0.1～0.35, water surplus, pH value are 7.0.The culture medium prescription of mycobacterium graminis is (weight %) in fermentation tank: sucrose 2～3, and peptone 0.5～0.8, diammonium phosphate 0.15～0.4, dipotassium hydrogen phosphate 0.05～0.1, water surplus, pH value are 7.0.

The present invention attempts to produce various nutrient elements from the fermentation of microorganism mycobacterium graminis, improves people's health level by ecosystem cure, thereby following advantage is arranged:

One. by the synergism of above-mentioned multiple components, promote human body greatly, and strengthen physiologically active for its absorption rate.Lipid-antioxidant activity experimental results show that: V _CMix use than using its oxidation resistance to increase more than ten times separately with flavone.

They are two years old. and this immune liquid contains mycobacterium muramyldipeptide polysaccharide MDP, it is an immunocompetence unit minimum in the cell wall, and the poly-polysaccharide of arabinose galactose is an immunizing antigen, very promote the generation and the release of cells in vivo active factors with producing effect, as: the generation of tumor necrosis factor, interferon, interleukin etc., obviously improve SOD enzymatic activity in the liver homogenate, reduce the content of LPO in the serum; Activating macrophage and NK cell improve the immunity of human body greatly, life cycle and the survival rate of the disposable bigger metering irradiation mice of significant prolongation, and numeration of leukocyte is also apparently higher than matched group.

They are three years old. played diseases prevention, anticancer effect: obvious inhibitory action is arranged for herpesvirus; Gastrointestinal function disorder and ulcer, diabetes, asthma all there is positive effect; Even in various degree alleviation and healing effect are also arranged for multiple cancer.The leukopenic patient who causes by chemicotherapy, take this biological product, 20ml * 2/ day or 1 gram/sky (powder) are through in a fortnight, leukocyte rises to 6000 by 2000, early stage osteocarcinoma and gastric cancer are all had the healing effect, and even late period, liver, pulmonary carcinoma also there was the case that delays vital stage of 50%.

They are four years old. with V _C, V _E, carotene, flavone four big antioxidant system concentrates in the system, thereby avoided the murder by poisoning of free radical and singlet oxygen effectively.

They are five years old. and the multiple effective active factor in this product exists with the state of trace or ultramicron, and is not only effective, and without any side effects.

They are six years old. and technology is simple, and is easy to operate.

Description of drawings

Accompanying drawing is a technological process block diagram of the present invention.

The specific embodiment

One. the preparation of Mackefulan immune liquid

Comprise: adopt genetic engineering method, microorganism (strain) sq-1 that selection-breeding one strain is good, promptly preserving number is the mycobacterium graminis (Mycobaterium phlei) of CGMCCNO.0916, is placed in the seed tank and cultivates; In fermentation tank, cultivate and ferment, obtain to contain the biological product of carotene, flavone multivitamin and muramyldipeptide polysaccharide; By sparing matter and sterilization, operation, make Mackefulan immune liquid product again.

In seed tank, the culture medium prescription of mycobacterium graminis is (weight %): sucrose 1.5～2.8, and Semen Maydis pulp 0.4～0.8, diammonium phosphate 0.15～0.4, dipotassium hydrogen phosphate 0.05～0.1, yeast extract 0.1～0.35, water surplus, pH value are 7.0.The culture medium prescription of mycobacterium graminis is (weight %) in fermentation tank: sucrose 2～3, and peptone 0.5～0.8, diammonium phosphate 0.15～0.4, dipotassium hydrogen phosphate 0.05～0.1, water surplus, pH value are 7.0.

The pulverizing of above-mentioned biological product can be carried out the high pressure draught cutting by high pressure homogenizer.The product that high pressure homogenizer adopts DenmarkAPV company to produce.

Be described further below in conjunction with the technology of accompanying drawing the preparation of Mackefulan immune liquid.

1. seed culture: adopt genetic engineering to cultivate the mycobacterium graminis bacterial strain, with its microscopy and sampling chemical examination, therefrom good microorganism (strain) sq-1 of selection-breeding one strain, i.e. mycobacterium graminis (Mycobaterium phlei) CGMCC NO.0916.

2. prepare burden: press table one, the listed composition of raw materials of table two, under filtrated air, steam and defoamer condition, allot culture medium used in seed tank, the fermentation tank.Wherein, adopt the water of handling through water correction plant.When seed tank, ferment tank, should remain under filtrated air, cooling water and the steam condition.

3. seed: raw materials mixed is put into seed tank together with the mycobacterium graminis seed liquor of being cultivated cultivate.

Process conditions are: 28～30 ℃ of temperature, 15～20 hours time, pH value is 7.0.

4. fermentation: the mycobacterium graminis bacterial strain of institute's selection-breeding is put into fermentation tank cultivate and ferment.Process conditions are: 28～30 ℃ of temperature, through 40 hours aerobic culture, pH value was 7.0 at least.

Goods in the above-mentioned operation 3,4, before flowing into next procedure, should be with its microscopy and sampling chemical examination.

5. even matter: with the product of fermentation tank, carry out the high pressure draught cutting, get nanoscale homogenate goods by high pressure homogenizer.

6. aseptic subpackaged: that goods are put into by the bottle after wash bottle, the sterilization.

7. pack: comprise labeling, box-packed, outer package operation.

8. through check, finished product is put in storage or is dispatched from the factory.

Two. make the dermopathic medicine of treatment with the Mackefulan immune liquid

First homogenate product dried with above-mentioned Mackefulan immune liquid, reuse edible oil lixiviate at least 1 hour, and make medicine for external use such as treatment herpes and skin injury.

Three. making the medicine of diseases such as treatment gastric ulcer, diabetes, asthma and multiple cancer with the Mackefulan immune liquid, can be dosage forms such as injection or capsule.

Four. subordinate list

1. culture medium prescription

Table one. the mycobacterium graminis culture medium prescription (weight %) in seed tank

Table two. in fermentation tank, the culture medium prescription of mycobacterium graminis (weight %)

2. mice is drunk the experimental data behind the Mackefulan immune liquid

Table three. significantly strengthen the inductive spleen lymphocyte proliferation ability of mice CONA (OD value)

Table four. significantly strengthen mice is brought out the DTH reaction to DNFB left and right sides ear weight difference (mg)

Table five. the serum hemolysin that obviously raises (antibody product)

Table six. obviously strengthen the huge cell of nibbling of mice and gulp down and nibble function

(1) gulps down and nibble percentage rate

(2) gulp down and nibble index

Table seven. significantly strengthen mice carbon and clean up function (gulp down and nibble index)

Table eight. significantly increase mice serum and 1% liver homogenate SOD enzymatic activity (Nu/me)

(1) serum index

(2) 1% liver homogenate SOD enzymatic activitys (Nu/me)

Table nine. significantly reduce mice LPO content

(1) serum (n mol)

(2) 1% liver homogenate (MDA/me)

Table ten. radiation-resisting functional

With ⁶⁰The even irradiation of the disposable whole body of Co gamma-rays (30 days)

(1) survival natural law

As seen from the above table, the survival rate of matched group and experimental group (%) is respectively 60,100.

(2) be subjected to according to lencocyte count after 24 days (* 109/L)

Claims

1. Mackefulan immune liquid, it is characterized in that its preparation process comprises: adopt genetic engineering method, microorganism (strain) sq-1 that selection-breeding one strain is good, promptly preserving number is the mycobacterium graminis (Mycobateriumphlei) of CGMCC NO.0916, is placed in the seed tank and cultivates; In fermentation tank, cultivate and ferment, obtain to contain the biological product of carotene, flavone multivitamin and muramyldipeptide polysaccharide; By sparing matter and sterilization, operation, make Mackefulan immune liquid product again,

In seed tank, the culture medium prescription of mycobacterium graminis is (weight %): sucrose 1.5～2.8, and Semen Maydis pulp 0.4～0.8, diammonium phosphate 0.15～0.4, dipotassium hydrogen phosphate 0.05～0.1, yeast extract 0.1～0.35, water surplus, pH value are 7.0,

The culture medium prescription of mycobacterium graminis is (weight %) in fermentation tank: sucrose 2～3, and peptone 0.5～0.8, diammonium phosphate 0.15～0.4, dipotassium hydrogen phosphate 0.05～0.1, water surplus, pH value are 7.0.

2. Mackefulan immune liquid according to claim 1 is characterized in that: cultivated in seed tank 15～20 hours.

3. Mackefulan immune liquid according to claim 1 is characterized in that: in fermentation tank at least through 40 hours aerobic culture.

4. Mackefulan immune liquid according to claim 1 is characterized in that the even matter of biological product, carries out the high pressure draught cutting by high pressure homogenizer.

5. Mackefulan immune liquid according to claim 4 is characterized in that adopting the high pressure homogenizer of Denmark APV company.

6. make the dermopathic medicine of treatment with the described Mackefulan immune liquid of claim 1, it is characterized in that: earlier biological product are spared matter, get the homogenate goods of Mackefulan immune liquid, drying, reuse edible oil lixiviate at least 1 hour forms the medicine for the treatment of herpes and skin injury.

7. make the medicine of treatment disease with the described Mackefulan immune liquid of claim 1, it is characterized in that: pharmaceutical formulation is injection or capsule.