CN108524925A - A kind of inactive yeast bacterium injection - Google Patents
A kind of inactive yeast bacterium injection Download PDFInfo
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- A61K36/06—Fungi, e.g. yeasts
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Abstract
The present invention relates to a kind of enhancing immune function, antibacterial and antiviral injections, belong to biomedicine field.The main component of this drug is inactive yeast bacterium, by the way that the saccharomycete lived inactivation to be prepared.Inactive yeast bacterium drug administration by injection can enhance body's immunity, and with antibacterial, antivirus action, can be used for a variety of diseases such as body's immunity caused by various bacteriosis, viral disease and a variety of causes of human or animal is low prevent, treat or auxiliary treatment, have broad application prospects.
Description
Technical field
The invention belongs to biomedicine field more particularly to a kind of enhancing immune function, antibacterial and antiviral injections.
Background technology
It has now been found that saccharomycete has enteron aisle balance is adjusted, promotes food conversion and improves body's immunity etc.
Good probiotic properties are used as feed addictive to be used for livestock and poultry cultivation more.There is research using yeast as feed addictive for increasing
Immune function (the patent publication No. of strong pig：CN106387398A).In addition, by yeast extract or its active ingredient (such as yeast
Polysaccharide, yeast manna oligosacchride, yeast beta-dextran) it is used to improve the more (bibliography [1] of research of body's immunity：
Work(is immunized to weanling pig in Molist F, Eerden E V, Parmentier H K, Vuorenmaa J. hydrolyzed yeast extracts
It can be read extensively with the influence feeds of growth performance, 2014,12：32. bibliography [2]：Ding Peng, Hu Guili, Liu Yongqiang, Song Zehe,
Fan Zhiyong, influence .2017 of the yeast manna oligosacchride of congratulating somebody on a happy occasion to pig chicken production performance and immune function, 5：24-26. bibliography
[3]：Zhu Yamin, Zhang Haibo, summer Changhong, Zhang Yan, research of the Zhang little Li yeast beta-dextrans to mouse immunity humidification
Food and fermentation science and technology, 2016,52 (2)：1-2.).In addition, sulfuric acid is made after being chemically modified yeast dextran in somebody
Change or carboxymethyl yeast dextran injection, the immune function (patent publication No. for enhancing animal：CN102028704A,
CN102048686A)。
In the past we have discovered that the lactic acid bacteria of inactivation has the administration of the animals ivs such as mouse, ox, pig, dog good
Immune-enhancing effect, antibacterial and it is antiviral the effects that, and declared the multinomial patent of invention of international and national (number of patent application be as follows：
PCT/CN2016/09844,201610809584.6,201610811028.2,201610810757.6,201610810758.0,
201610809345.0 201610809582.7,201610809583.1,201610809676.4,201610811285.6,
201610811770.3).Based on above research, it is presumed that injection is made to animal drug administration by injection in saccharomycete inactivation
There should be immunological enhancement, be consulted and found by document and patent database, there is no saccharomycete directly goes out so at present
Report living as injection, we have discovered that being administered saccharomycete inactivation animal with good immune as injection
Enhancing and antibacterial, antivirus action.The drug also has the advantages such as at low cost, drug residue free to animal-use drug, especially to family
Poultry control and prevention of disease has broad application prospects.
Invention content
The main component of a kind of enhancing immune function, antibacterial and antiviral injection, the injection is inactive yeast bacterium,
Gram's staining is carried out to the inactive yeast bacterium in injection, in oily microscopic observation, inactive yeast bacterium keeps complete thalli morphology,
Thalline profile of viable bacteria and form are consistent before being inactivated with it.
It is 10 that the injection, which includes inactive yeast bacterium number amount per ml injections,5—1012It is a.
The saccharomycete is selected from saccharomyces cerevisiae category, torulaspora delbrueckii category, candida, Brunswick Durham yeast
Category, pichia, cloth Laplace saccharomyces, Torulopsis candida category, Xue watt saccharomyces, rhodothece rubra category, schizosaccharomyces pombe category,
Boydii saccharomyces.
Further, the ablation method of saccharomycete is selected from high-temperature inactivation, high temperature and pressure inactivation, ultraviolet inactivation, chemical reagent
Any one of inactivation or radiological inactivation.
Further, the injection also includes pharmaceutically acceptable adjuvant, contains enough salt or list in adjuvant
Sugar is same or similar to ensure injection suspension and colloidal osmotic pressure.
The dosage form of preparation inactive yeast bacterium injection for the present invention includes：Powder-injection, mixed suspension injection etc..Described
Powder-injection is made by being spray-dried or being freeze-dried, and suspension is made in when application.
The preparation is injection.
Preferably, the inactive yeast bacterium is single bacterium, carries out Gram's staining to inactive yeast bacterium, sees under the microscope
It examines, inactive yeast bacterium keeps complete thalli morphology, and thalline profile of viable bacteria and form are consistent before being inactivated with it, and drug gives prescription
Formula is drug administration by injection.
Further, the inactive yeast bacterium is two or more inactive yeast bacterium mixtures, and it is blue to carry out leather to inactive yeast bacterium
Albert'stain Albert is observed under the microscope, predominantly keeps the inactive yeast bacterium of complete thalli morphology, the administering mode of drug is injection
Administration.It is obtained using any one of high temperature and pressure inactivation, ultraviolet inactivation, chemical reagent inactivation or radiological inactivation ablation method
The inactive yeast bacterium.The saccharomycete is selected from saccharomyces cerevisiae category, torulaspora delbrueckii category, candida, Brunswick
Durham saccharomyces, pichia, cloth Laplace saccharomyces, Torulopsis candida category, Xue watt saccharomyces, rhodothece rubra category, grain wine are split
Grow saccharomyces, Boydii saccharomyces.
The adjuvant of the injection is selected from pungent phenoxy polyethoxyethanol, alevaire, sorbitol anhydride polyethyleneglycol
Oleate, polyoxyl 40 stearate, polyoxyethylene deriv, Tween-80, polyoxyethylene alkylphenyl sodium sulfonate, lauryl sulfate ester
Sodium, polysaccharide sulfate alkali metal salt, dextran sulfate sodium, sulfosuccinic acid double monooctyl esters, Arabic latex, gather acacin
Vinylpyrrolidone, ethyl polysilicate, ethyl alcohol, glycerine, sorbierite, honey, agar, starch, dextrose, fructose, malt extract,
Cocoa power, tartaric acid, citric acid, sodium citrate, angle fork acid, alginic acid, sodium alginate, tannic acid, cyclohexane sulfamic acid, mineral
Oil, eucalin, saccharin sodium, ghatti gum, karaya gum, tragacanth, pectin, antler glue, gelatin, carboxymethyl cellulose
Element, sulfate cellulose, methylcellulose, sodium carboxymethylcellulose, acetic acid sodium cellulose sulfate, sodium hydroxyethyl cellulose, methyl
Poly- silicon, potassium sorbate, kaolin, diatomite, bentonite, alumina silicate, aluminium hydroxide, colloidal aluminium hydroxide, aluminium-magnesium silicate, illiteracy are de-
Native magnesium, trimerization magnesium silicate, aluminum magnesium silicate sodium, sodium bicarbonate, sodium carbonate, methyl p-hydroxybenzoate, propylparaben,
Ethyl vanillin, lemon oil, orange oil, vanillic aldehyde, casein etc..
Preferably, saccharomycete of the present invention is selected from：Saccharomyces cerevisiae (latin name：Saccharomyces
Cerevisiae is purchased from Chinese industrial Microbiological Culture Collection administrative center；Deposit number：CICC1012), Dare is furnished with spore circle
Yeast (latin name：Torulaspora delbrueckii are purchased from Chinese industrial Microbiological Culture Collection administrative center；Preservation
Number：CICC1004).Respectively to above 2 Yeasts carry out conventional method inactivation, then to mouse vein be administered, find with
Upper 2 kinds of inactive yeast bacterium intravenously administrable can enhance mouse body's immunity, and have antibacterial and antivirus action.
Inactive yeast bacterium injection of the present invention can be used for the various bacteriosis of human or animal, viral disease
A variety of diseases such as body's immunity caused by disease, fungal disease, parasitic disease, cancer and a variety of causes is low it is pre-
Anti-, treatment or auxiliary treatment.
Nobody attempts with inactive yeast bacterium drug administration by injection for improving body's immunity, antibacterial and antiviral before this
Research, may mainly considering inactive yeast bacterium drug administration by injection, there are safety issues, with water-soluble intravenous injection used in everyday
Agent compares, and inactive yeast bacterium injection is in graininess suspension, such particulate matter is carried out drug administration by injection, it is believed that wind
Danger is higher.But current people doctor has clinically had the drug that the thalline of harmful bacteria is carried out to drug administration by injection, and such as BCG vaccine is tuberculosis
Live vaccine can be used to prevent the auxiliary treatment of tuberculosis and tumour by oral, subcutaneous, abdominal cavity or intratumor injection.It is short and small rodlike
Bacillus preparation is the dead bacteria suspension of corynebacterium, through in subcutaneous, muscle, tumor or intravenous drip etc. is for some tumours
Auxiliary treatment.In addition, also A group streptococcus preparation, P. aeruginosa bacteria preparation, Mycobacterium graminis preparation etc..In addition, somebody
By the complete thin of Rhod, Gordona, Nocardia, enlightening thatch Bordetella, tomb village Bordetella and class Nocardia
The administration of bacterium cell infusion is used as immunomodulator (patent publication No.：CN1735431A).
And the present invention pass through the study found that inactive yeast bacterium intravenous injection have improve immune function and prevention bacterial disease,
The effect of virosis.Saccharomycete is inactivated drug administration by injection as a kind of probiotics has safer characteristic.Injection inactivation ferment
The mechanism that female bacterium can improve body's immunity may be after inactive yeast bacterium drug administration by injection, and body is by inactive yeast bacterium
Thalline is identified as foreign substance, in order to be disposed, to enhance body's immunity；It is also possible to be to maintain complete
The inactive yeast bacterium of thalli morphology contacts with immunocyte, acts on Toll-like receptor on immunocyte, to activate siberian crabapple
System.Inactive yeast bacterium drug administration by injection have antibacterial and antivirus action may be enhance body's immunity as a result, also having
It may be the effect that the substance generated after body catabolism after inactive yeast bacterium drug administration by injection plays.
Description of the drawings
The saccharomyces cerevisiae Gram's staining oil mirror that Fig. 1 lives observes photo
Fig. 2 inactivates saccharomyces cerevisiae Gram's staining oil mirror and observes photo
Specific implementation mode
Embodiment 1
Saccharomyces cerevisiae (is purchased from Chinese industrial Microbiological Culture Collection administrative center；Latin name：Saccharomyces
Cerevisiae, deposit number：CICC1012) be inoculated in culture medium (weigh 20g peptones, 20g glucose, 10g yeast powders,
1000mL is added water to, prepared by 121 DEG C of sterilizing 20min), it is small with the rotating speed culture 24 of 160r/min in 37 DEG C of constant temperature oscillators
When, then 3000 leave the heart 5 minutes, removal culture supernatants retain precipitation, and sterile saline cleaning precipitation, centrifugation 5 is added
Minute, after repeated washing 3 times, sterile saline is added, with precipitation mixing, suspension is made.Take a certain amount of saccharomyces cerevisiae
Bacterium normal saline suspension carries out count of bacteria by THOMA bateria chambers, every 1ml suspensions is made to contain 109A wine brewing ferment
Female thalline.By prepared saccharomyces cerevisiae normal saline suspension in 121 DEG C, pressure 0.12MPa, 15min is inactivated, is inactivated
Saccharomyces cerevisiae injection.Saccharomyces cerevisiae living and the saccharomyces cerevisiae of inactivation are subjected to Gram's staining respectively, in oily microscopic observation
Thalli morphology (referring to attached drawing 1 and attached drawing 2), comparison discovery, the thalline profile and form that inactivated bacteria is consistent with viable bacteria, bacterium
It counts and finds that significant change does not occur for the front and back thalline quantity of inactivation.Inactivation saccharomyces cerevisiae normal saline suspension is centrifuged,
It abandons supernatant and stays precipitation, extract DNA, using PCR amplification 16S rDNA, saccharomycete type discriminating can also be carried out by sequencing.
Embodiment 2
By torulaspora delbrueckii, (torulaspora delbrueckii is purchased from the management of Chinese industrial Microbiological Culture Collection
The heart；Latin name：Torulaspora delbrueckii, deposit number：CICC1004 it) is inoculated in culture medium and (weighs 20g eggs
White peptone, 20g glucose, 10g yeast powders add water to 1000mL, prepared by 121 DEG C of sterilizing 20min), in 37 DEG C of constant temperature oscillators
With the rotating speed culture 24 hours of 160r/min, then 3000 the heart is left 5 minutes, removal culture supernatants retain precipitation, and nothing is added
Bacterium physiological saline cleaning precipitation, centrifuges 5 minutes, and after repeated washing 3 times, sterile saline is added, and with precipitation mixing, is made mixed
Suspension.A certain amount of torulaspora delbrueckii bacterium normal saline suspension is taken, bacterium meter is carried out by THOMA bateria chambers
Number, makes every ml suspensions contain 109A torulaspora delbrueckii thalline.By prepared torulaspora delbrueckii bacterium physiology
Salt aqueous suspension inactivates 15min in 121 DEG C, pressure 0.12MPa, obtains inactivation torulaspora delbrueckii bacterium injection.
Embodiment 3
Detect the inactivation saccharomyces cerevisiae injection and embodiment 2 of the preparation of embodiment 1 respectively using carbon particle clearance test method
Influence of the inactivation torulaspora delbrueckii injection of preparation to normal mouse monocytes/macrophages phagocytic function.By weight
Cleaning grade Kun Ming mice for 18-22g, which is divided into Normal group, inactivation saccharomyces cerevisiae injection group and inactivates Dare, to be furnished with
Spore torula injection group.Every group of 10 mouse, half male and half female.Normal group mouse tail vein injection sterile saline,
It inactivates saccharomyces cerevisiae injection group mouse tail vein injection and inactivates saccharomyces cerevisiae suspension, inactivation torulaspora delbrueckii injection
Agent group mouse tail vein injection inactivates torulaspora delbrueckii suspension.The above each group administered volume is 0.1mL/10g, even
Continuous tail vein injection 5 days, 1 time a day.After last dose 2 hours, mouse tail vein injection india ink 0.05mL/10g, in
1min and 10min from orbital venous plexus 40 μ L of blood sampling, is added to 4mL 0.1%Na respectively2CO3It is shaken up in solution, uses spectrophotometer
The colorimetric under 680nm wavelength measures optical density and (uses OD individually below1And OD10To indicate the light of 1min and 10min institutes blood sampling
Density), carbonic clearance index K values are calculated as follows.Carbonic clearance index K=(lgOD1-lgOD10)/(t10-t1).Using SPSS
11.5 softwares carry out significance test to experimental data, the results are shown in Table 1.As shown in Table 1, compared with Normal group, inactivation
Saccharomyces cerevisiae injection group and inactivation torulaspora delbrueckii injection group significantly significantly improve carbonic clearance and refer to pole respectively
Number K values.Described above, will inactivate saccharomyces cerevisiae and torulaspora delbrueckii intravenously administrable can improve the list of normal mouse
Core-macrophage phagocytic function improves the non-specific immune function of mouse.
1 inactive yeast bacterium of table is injected intravenously the influence result to normal mouse monocytes/macrophages phagocytic function
Note：* indicates to indicate and Normal group comparing difference with the poor heteropolar notable P ＜ 0.01 of Normal group, *
Notable P ＜ 0.05.
Embodiment 4
Be respectively adopted iodine [125I] tumor necrosis factor-alpha radioimmunoassay kit and iodine [125I] interferon-r radiation
Inactivation saccharomyces cerevisiae injection prepared by immunoassay kits detection embodiment 1 contains TNF-α in mice serum and IFN-γ
The influence of amount.Significance test is carried out to experimental data using 11.5 softwares of SPSS, the results are shown in Table 2.As shown in Table 2, with it is normal
Control group compares, and inactivation saccharomyces cerevisiae injection group pole significantly improves TNF-α and IFN-γ content in mice serum.
2 inactive yeast bacterium of table is injected intravenously the influence result to TNF-α in mice serum and IFN-γ content
Note：* indicates to indicate and Normal group comparing difference with the poor heteropolar notable P ＜ 0.01 of Normal group, *
Notable P ＜ 0.05.
Embodiment 5
Inactivation Dare prepared by inactivation saccharomyces cerevisiae injection and embodiment 2 prepared by detection embodiment 1 is furnished with spore circle ferment
Preventive and therapeutic effect of female injection to salmonella infection mouse.The cleaning grade Kun Ming mice that weight is 18-22g is divided into 4
Group, i.e. Normal group, salmonella group, inactivation saccharomyces cerevisiae injection group and inactivation torulaspora delbrueckii injection
Group, every group of 30 mouse, half male and half female.The mouse tail vein injection sterile saline of Normal group and salmonella group,
It is 0.1mL/ to inactivate saccharomyces cerevisiae injection group and inactivation torulaspora delbrueckii injection group difference tail vein injection dosage
The corresponding injections of 10g.It is spaced 24 hours after administration, salmonella group, inactivation saccharomyces cerevisiae injection group and inactivation Dare are furnished with
The mouse difference tail vein injection Bacterium enteritidis of spore torula injection group (is purchased from Chinese veterinary microorganism culture presevation pipe
Reason center, number CVCC 3377).It is observed continuously 20 days, the daily death toll of record mouse.Using SPSS11.5 softwares to reality
It tests data and carries out Chi-square Test, the results are shown in Table 3.As shown in Table 3, compared with salmonella control group, inactivation saccharomyces cerevisiae injection
Agent group and inactivation torulaspora delbrueckii injection group pole significantly reduce the death toll of mouse.Illustrate inactivation wine brewing ferment
Female injection and inactivation torulaspora delbrueckii injection intravenously administrable have preventive and therapeutic effect to the mouse for infecting salmonella.
The result that 3 inactive yeast bacterium of table intravenous injection prevention salmonella acts on mouse lethal
Note：* indicates often to organize with normal control poor with the poor heteropolar notable P ＜ 0.01 of Normal group, * expressions
Different notable P ＜ 0.05；△ △ indicate to indicate and salmonella group ratio with the poor heteropolar notable P ＜ 0.01 of salmonella group, △
Compared with significant difference P ＜ 0.05.
Embodiment 6
Inactivation Dare prepared by inactivation saccharomyces cerevisiae injection and embodiment 2 prepared by detection embodiment 1 is furnished with spore circle ferment
The influence of female PR8 plants of infecting mouse death toll of yeast injection agent infected by influenza.Experiment selects 18-22g cleaning grades Kunming kind small
White mouse is divided into Normal group, PR8 plant model group (i.e. influenza virus PR8 plants of infecting mouse model groups), inactivation saccharomyces cerevisiae note
Penetrate agent group and inactivation torulaspora delbrueckii injection group.Every group of 30 mouse inactivate saccharomyces cerevisiae injection group and inactivation
Torulaspora delbrueckii injection group distinguishes the corresponding injection that tail vein injection volume is 0.1ml/10g；Normal group
With PR8 plants of model group tail vein injection sterile salines, each group administered volume is 0.1mL/10g.Each group mouse is in administration 1
Interval 24 hours after secondary, in addition to Normal group, other each group mouse contain influenza under ether light anesthesia in intranasal vaccination
Viral PR8 plants of chick embryo allantoic liquid, every 0.05mL make PR8 plants of infecting mouses of influenza virus.Hereafter it observes and records each in 10 days
Group dead mouse number.Chi-square Test is carried out to experimental data using 11.5 softwares of SPSS, the results are shown in Table 4.As shown in Table 4, it inactivates
Saccharomyces cerevisiae injection group and inactivation torulaspora delbrueckii injection group with PR8 plant model groups compared with, respectively it is extremely notable with
Significantly reduce dead mouse number.Inactivation saccharomyces cerevisiae described above and inactivation torulaspora delbrueckii intravenously administrable can be used
It is prevented in viral disease.
4 inactive yeast bacterium of table is injected intravenously the influence result of PR8 plants of infecting mouses of infected by influenza
Note：* indicates often to organize with normal control poor with the poor heteropolar notable P ＜ 0.01 of Normal group, * expressions
Different notable P ＜ 0.05；△ △ indicate to indicate and PR8 plants of model groups with the poor heteropolar notable P ＜ 0.01 of PR8 plants of model groups, △
The notable P ＜ of comparing difference 0.05.
Claims (9)
1. the main component of a kind of enhancing immune function, antibacterial and antiviral injection, the injection is inactive yeast bacterium,
It is characterized in that, carry out Gram's staining to the inactive yeast bacterium, in oily microscopic observation, inactive yeast bacterium keeps complete thalline
It is 10 that form, which includes inactive yeast bacterium number amount per ml injections,5—1012A, the injection is suspension, and administering mode is note
Penetrate administration.
2. injection as described in claim 1, which is characterized in that the saccharomycete is saccharomyces cerevisiae (latin name：
Saccharomyces cerevisiae, deposit number：CICC1012).
3. injection as described in claim 1, which is characterized in that the saccharomycete is torulaspora delbrueckii (Latin
Title：Torulaspora delbrueckii, deposit number：CICC1004).
4. injection as described in claim 1, which is characterized in that the saccharomycete is selected from saccharomyces cerevisiae category, Dare is furnished with
Spore torulopsis, candida, Brunswick Durham saccharomyces, pichia, cloth Laplace saccharomyces, Torulopsis candida category, Xue
It is one or more in watt saccharomyces, rhodothece rubra category, schizosaccharomyces pombe category, Boydii saccharomyces.
5. injection according to claim 1, which is characterized in that ablation method is selected from high-temperature inactivation, high temperature and pressure inactivates,
Any one of ultraviolet inactivation, chemical reagent inactivation or radiological inactivation.
6. injection according to claim 1, which is characterized in that the injection also includes pharmaceutically acceptable assistant
Agent, containing enough salt or monosaccharide to ensure that injection suspension and colloidal osmotic pressure are same or similar in adjuvant.
7. injection according to claim 1, which is characterized in that the injection is by being spray-dried or being freeze-dried
It is fabricated to powder-injection, when application is configured to suspension.
8. according to claim 1-7 any one of them injections, which is characterized in that the injection can be with intravenously administrable, skin
Lower injection, intracutaneous injection, intramuscular injection, intraperitoneal injection, intratumor injection administration.
9. purposes of the injection according to claims 1-8 in preparing enhancing immune function, antibacterial and antiviral drugs.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112076312A (en) * | 2019-06-13 | 2020-12-15 | 石小和 | Vaccine with immunoregulation function, multiple antiserum injection and preparation method |
| WO2022068924A1 (en) | 2020-09-30 | 2022-04-07 | 成都夸常奥普医疗科技有限公司 | Use of probiotic component and pharmaceutical composition containing probiotic component |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103966111A (en) * | 2009-01-27 | 2014-08-06 | 乐斯福公司 | Saccharomyces cerevisiae strains with phytosanitary capabilities |
| CN105131146A (en) * | 2015-08-18 | 2015-12-09 | 陈莉 | Combined extraction of beta-glucan and beta-mannan in yeast cell walls |
-
2018
- 2018-04-20 CN CN201810359624.0A patent/CN108524925B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103966111A (en) * | 2009-01-27 | 2014-08-06 | 乐斯福公司 | Saccharomyces cerevisiae strains with phytosanitary capabilities |
| CN105131146A (en) * | 2015-08-18 | 2015-12-09 | 陈莉 | Combined extraction of beta-glucan and beta-mannan in yeast cell walls |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112076312A (en) * | 2019-06-13 | 2020-12-15 | 石小和 | Vaccine with immunoregulation function, multiple antiserum injection and preparation method |
| WO2022068924A1 (en) | 2020-09-30 | 2022-04-07 | 成都夸常奥普医疗科技有限公司 | Use of probiotic component and pharmaceutical composition containing probiotic component |
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| CN108524925B (en) | 2020-12-01 |
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