CN101919878A - Application of coryebactrium parvum in preparing medicament for treating novel A H1N1 influenza viruses - Google Patents
Application of coryebactrium parvum in preparing medicament for treating novel A H1N1 influenza viruses Download PDFInfo
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Abstract
The invention relates to application of a coryebactrium parvum preparation (CPP) or a noncellular coryebactrium parvum preparation (NCP) in the field of pharmacy, in particular to application in preparing a medicament for treating novel A H1N1 influenza viruses. The coryebactrium parvum preparation is a preparation prepared by inactivating coryebactrium parvum through formaldehyde; and the noncellular coryebactrium parvum preparation is of a nanometer grade and has uniform grain size, enhanced absorbance and diffusance and low heat source without formaldehyde residuals. The CPP plays a certain role of resisting the novel A H1N1 influenza viruses, and especially, the effect of the NCP is more obvious.
Description
Technical field
The present invention relates to a kind of short corynebacteria preparation (CPP) or the application of acellular short corynebacteria preparation (NCP) in pharmaceutical field, relate in particular to the application in the novel influenza A H1N1 influenza virus medicament of preparation treatment, belong to drug world.
Background technology
Influenza be a kind of by influenza virus cause, infectious disease by respiratory infectious.Being very popular of influenza once caused very serious consequence to the mankind in history, " spanish influenza " that cause by H1N1 subtype influenza virus that for example broke out in 1918, be exactly the catastrophic influenzas of tool of 20th century, infect half population of the whole world in less than one-year age, caused tens of millions of people's death.Flu outbreak is normally by a kind of emerging or cause with the virus that forefathers had not infected, human body does not have immunity to this new virus, add that its spread scope is wide, speed is fast, and the development of vaccine and production are difficult to catch up with immediately, so cause many people to infect easily even death.In recent years, continuous intensification along with the integrated degree of global economy development, people-to-people contacts between the countries in the world and mobile frequent day by day and close, thereby influenza virus is further accelerated in the speed of sending out and spreading in the whole world, in case flu outbreak takes place in certain country, epidemic situation will be sent out rapidly to all over the world at short notice.The Type A Influenza H1N1 epidemic situation that took place in 2009 just so.
In April, 2009, the first Type A Influenza H1N1 case has been made a definite diagnosis by Mexico.During April to May subsequently, the Type A Influenza H1N1 epidemic situation is rapid spread worldwide, and only in the time of a wheat harvesting period, the Type A Influenza H1N1 epidemic situation has all taken place in global a plurality of countries and regions.Mexico and the U.S. the most serious with epidemic situation are example, and by on May 14th, 2009, Mexico made a definite diagnosis in the whole nation influenza A H1N1 case and rises to 2720 examples, and wherein death toll rises to 64 people; And in 50 states of the U.S., the influenza A H1N1 case is announced to find in existing 47 states, and patient's sum also rises to 4298 people, increases by 946 people than the previous day.The first influenza A H1N1 case was also made a definite diagnosis on ground in China May 11, by on May 19th, 2009, occurred 4 routine influenza A H1N1 cases altogether, was Introduced cases influenza case.World Health Organization (WHO) announces that on April 29th, 2009 global flu outbreak warning level is 5 grades, and along with influenza A H1N1 epidemic situation constantly spreading worldwide, the risk of breaking out global flu outbreak promotes day by day, World Health Organization (WHO) has clearly represented not get rid of at the beginning of 5 months warning level is risen to 6 grades, promptly announces the probability that flu outbreak arrives.
After this time the Type A Influenza H1N1 epidemic situation took place, the scientists of countries in the world had been launched relevant scientific research immediately.Up-to-date investigation and result of study show that each Mexico patient who infects novel influenza A H1N1 can cause increasing by 1.2 to 1.6 new cases, and the interpersonal communication who proves influenza virus is in continuous generation.Up to the present the infectious rate of this influenza virus is 1/3, may have the infection that 2,000,000,000 people are subjected to this influenza virus in the global range, and global big influenza is first meeting clue now.But owing to still can not determine at present the origin of toxicity, propagability and the virus of novel influenza A H1N1 influenza virus, and in should virus incubation period and infective stage, neither be very clear and definite, so scientists also can't be judged the degree that it worldwide spreads.
Summary of the invention
The object of the present invention is to provide a kind of short corynebacteria preparation (CPP) or the new application of acellular short corynebacteria preparation (NCP) in pharmacy, i.e. application in the novel H 1 N 1 influenza A virus infection of preparation treatment, described short corynebacteria preparation is the preparation of being made behind formalin-inactivated by short corynebacteria (CP), described NCP is a nanoscale, the granular size homogeneous, trap and diffusance improve, low pyrogen, formaldehydeless residual, have identical spleen activation with the whole cell preparation and reach biologic activity such as pressing down tumor, can reduce the fever of CPP, chest pain, the injection site redness, scleroma, side effect such as Liver and kidney toxicity; Short corynebacteria preparation (CPP) or acellular short corynebacteria preparation (NCP) all have tangible antiviral effect as a kind of nonspecific immunity regulator, can be used for the treatment or the auxiliary treatment of novel H 1 N 1 influenza A virus infection.
To achieve these goals, the technical solution used in the present invention is: the application of a kind of short corynebacteria preparation in the novel influenza A H1N1 influenza virus medicament of preparation treatment.
The application of a kind of acellular short corynebacteria preparation in the novel influenza A H1N1 influenza virus medicament of preparation treatment.
Short corynebacteria preparation of the present invention (CPP, 2000 editions " Chinese biological goods rules ") is a kind of nonspecific immunity regulator.It is the bacterin of being made behind formalin-inactivated by short corynebacteria (CP), and its preparation process is disclosed in 1995 editions and 2000 editions " Chinese biological goods rules ".The CPP preparation of using also has the inactivated vaccine that French Merleux institute is produced at present; The inactivated vaccine that Britain Wellcome pharmaceutical factory produces etc.
CPP has the ability of activated mononuclear phagocyte system.CPP mainly shows immune effect: the activated mononuclear macrophage, make the NK cytoactive strengthen, promote that body produces IgG and IgM, lures dirt interferon, interleukin II etc. multiple antigen, therefore have antitumor and anti-infectious function.
The antitumor action of CPP is confirmed by many experiments and clinical research.To using CPP before and after the laboratory animal inoculated tumour, can significantly suppress some solid tumor (as breast carcinoma, melanoma etc.), ascitic type tumor and leukemic growth.The animal of tumor regression has specific resistance for the tumor cell of attacking once more.CPP also has inhibitory action to neoplasm metastasis.
CPP antitumor and anti-infective core are by activating macrophage, and its quality and quantity is improved greatly, and activated macrophage can kill the microorganism of tumor cell and infection.
CPP is used for the history of the existing many decades of tumor and other treatment of diseases as immunomodulator, and it is generally acknowledged by activated mononuclear macrophage system antitumor, anti-infective effect.
02123571.6,02123570.8 and 02123572.4 applied for a kind of acellular short corynebacteria preparation, its preparation method and the purposes on preparation treatment tuberculosis medicine.
Acellular short corynebacteria preparation of the present invention (NCP) is that granularity is less than 100nm by the cell-free preparation of short corynebacteria (CP) through the fragmentation preparation.
Wherein said CP, formal name used at school are that propionibacterium acne (propionibacterium acnes) bacterium number is 65101 (76-27 or 7627), 65102 (H-84) or 65103 (77-1 or 771).
The pyrogen of described NCP is less than 160EU/ml, protein content 10～40% (g/g), and nucleic acid content 3～25% (g/g), all the other are polysaccharide.
The pyrogen of preferred NCP of the present invention is less than 60EU/ml, protein content 20～30% (g/g), and nucleic acid content 3～10% (g/g), all the other are polysaccharide.
The granularity of NCP of the present invention is preferably 50～80nm.
NCP of the present invention prepares by following step:
1) CP work seed lot strain is continuous is inoculated in big bottle or nutrition jar after going down to posterity for 2～4 times, cultivates in culture medium 5～7 days:
2) thalline of the no living contaminants of collection, heated and boiled obtained bacterium liquid in 15～60 minutes:
3) the bacterium liquid that sterility test is qualified is after washing, with the broken thalline of breaker;
4) collecting precipitation after centrifugal of the suspension behind the bacterial cell disruption, suspension is made in washing precipitation:
5) with the sterilization of suspension packing post-heating, obtain acellular short corynebacteria preparation.
Wherein the CP formal name used at school described in the step 1) is a propionibacterium acne, and bacterium number is 65101 (76-27 or 7627), 65102 (H-84) or 65103 (77-1 or 771).
Culture medium is to be selected from a kind of in THIOGLYCOLLIC ACID salt, peptone, liver or the yeast dialysis solution culture medium, and is low pyrogen culture medium.
Condition of culture is 36～37 ℃ of cultivations in the step 1).
Preferred described culture medium is not for containing the sulphur glycollate culture medium of agar.
Wherein the thalline described in the step 3) is an inactivated bacteria, and cell concentration is 50～50,000,000,000/ml.
Described breaker is that bacterial cell disruption is arrived less than 100nm, and preferred size is the superhigh pressure water jet collider of 50～80nm.
Wherein said bacterial cell disruption is to carry out under aseptic condition.
Wherein the washing described in the step 3) be with precipitate with physiological saline solution centrifuge washing at least 2 times: the washing described in the step 4) is with physiological saline solution centrifuge washing 0～5 time with precipitate.
The centrifugal condition of suspension is centrifugal 30～150 minutes of 4 ℃～room temperature, 6000～12000rpm behind the step 4) bacterial cell disruption.
Being heated to be described in the step 5) 60～65 ℃ of heating 0.5～2 hour.
Culture medium described in the present invention is preferably the low pyrogen culture medium through 0.22 μ m or 0.45 μ m membrane filtration.
In the preparation method of the present invention, also comprise the NCP lyophilization that step 5) is obtained, obtain the lyophilized formulations of NCP.
The NCP pyrogen of method for preparing is less than 160EU/ml, protein content 10～40% (g/g), and nucleic acid content 3～25% (g/g), all the other are polysaccharide.
The NCP pyrogen of preferred method for preparing is less than 60EU/ml, protein content 20～30% (g/g), and nucleic acid content 3～10% (g/g), all the other are polysaccharide.
The NCP granularity of method for preparing is less than 100nm, and preferred size is 10-50nm.
Steriling test described in the present invention is qualified to be meant must not check according to the method for " Chinese biological goods rules " regulation any bacterial growth.
Hang down pyrogen culture medium, particularly sulphur glycollate culture medium owing to adopting among the present invention, and in conjunction with strict Quality Control, and aseptic low grade fever origin operation in the whole process of production, guaranteed that final products hang down the pollution of pyrogen and the assorted bacterium of nothing.
The present invention is crushed to nanoscale by the superhigh pressure water jet collider with CP, by selecting different centrifugal force (rotating speed), determined centrifugal extraction conditions, the precipitation that obtains partly is NCP, prove through animal experiment repeatedly, said preparation has identical spleen activation with whole cell and reaches biologic activity such as pressing down tumor, and supernatant part then alienia activates and tumor-inhibiting action, shows that NCP of the present invention has good biologic activity.
Removing residue formaldehyde especially to the zest of responsive pleura, adopts heat inactivation to substitute formalin-inactivated to the zest of body among the CPP among the present invention in order to eliminate.Zoopery shows that heat-killed NCP, CPP have identical spleen activation and but biologic activity such as tumor than the CPP of formalin-inactivated.
The operation principle of superhigh pressure water jet collider is that (operating pressure is 10-3000kgf/cm to the liquid that will be mixed with machined object with high-pressure pump
2) be divided into two strands after the pressurization, entering two diameters with high-speed jet is that flow out the head-on collision back in opposite directions in the passage formed of 8mm diamond wafer.Because the speed of liquid is very high, when clashing in opposite directions, fluid produces very strong shock wave, and make diamond wafer produce high frequency, high intense ultrasonic wave.The high-power high-frequency ultrasound wave makes the moment pulverizing of machined object granule, ultramicronising.
In the preparation process of the present invention, being broken thalline better, adopting the superhigh pressure water jet collider, is 50～50,000,000,000/ml at cell concentration, and operating pressure is 800～1500kgf/cm
2Condition under broken, thalline suspension after the fragmentation is under 4 ℃～room temperature, centrifugal 30～150 minutes of 6000～12000rpm, abandon supernatant, precipitation is made suspension with behind the physiological saline solution solution washing 0～5 time, through prepare, packing, heated 0.5～2 hour at 60～65 ℃, the qualified back of sterility test merges, and obtains the good purpose product of high-quality, granularity and homogeneity.
NCP of the present invention is that cell breakage with CP is to nanoscale, through the invalid supernatant part of centrifugal removal, collecting precipitation partly prepares, product has low pyrogen, no living contaminants, formaldehydeless residual, and granule has nanoscale, homogeneous grain diameter, trap and diffusance improve, help absorption of human body and improve drug effect: can reduce fever, chest pain simultaneously, side effect such as injection site redness, scleroma are a kind of promising immune formulations.As required, NCP of the present invention can make the suspension formulation of physiological saline solution, solids content is 1.0mg/ml or 2.0mg/ml, also can make freeze-dried powder or be fit to other medicinal dosage forms: CPP of the present invention can be dosage form commonly used at present, as suspension formulation, freeze-dried powder or suitable other medicinal dosage forms.
Used CPP, the NCP of the present invention is nonspecific immunity strengthening agent, NCP is by the cell-free preparation of CP through the fragmentation preparation, for nanoscale, granular size homogeneous, trap and diffusance improve, low pyrogen, formaldehydeless residual, have identical spleen activation with the whole cell preparation and reach biologic activity such as pressing down tumor, can reduce fever, the chest pain of CPP, injection site redness, scleroma, side effect such as Liver and kidney toxicity, therefore NCP has overcome the some shortcomings of CPP, and the nonspecific immunity strengthening agent that can be used as a kind of excellent performance is applied to treatment of diseases such as antitumor and infection.
CPP of the present invention, NCP are nonspecific immunity strengthening agent, mainly act on macrophage, mononuclear phagocyte system had powerful and persistent stimulation, and can be by promoting that hematopoietic pluripotential stem cell is divided into mononuclear phagocyte, transfer the potentiality that body immune system is contained, bring into play anticancer, microbicidel function.
The anti-novel influenza A H1N1 influenza virus effect of CPP, NCP mainly is by to the powerful and persistent stimulation of mononuclear phagocyte system, causes mononuclear phagocyte hypertrophy, activation, phagocytic function enhancing and secretion inducing interferon (IFN-γ), interleukin (IL-2) active oxygen (H
2O
2, NO) etc. killer factor and strengthen the NK cell killing activity, reach the purpose that suppresses and kill and wound novel influenza A H1N1 influenza virus.
Along with the development to pathogenetic deeply understanding of novel influenza A H1N1 influenza virus and molecular immunology theory, the effect of nonspecific immunity in anti-novel influenza A H1N1 influenza virus more and more paid attention to by people.The M phagocyte not only plays an important role in the body nonspecific immunity, and also plays important effect in specific immunity.Mononuclear phagocyte system is removed has phagocytic function, also can bring into play antigen presentation (antigenpresentation) effect, also belong to antigen-presenting cell (antigen-presentation cell, APC), but helper T lymphocyte activation, and stimulate its function mutually, thereby amplify the specific immunity effect with lymphocyte.Yet have only the anti-novel influenza A H1N1 influenza virus effect of activatory phagocyte competence exertion, and the anti-novel influenza A H1N1 influenza virus of activatory phagocyte is not subjected to novel influenza A H1N1 influenza virus variation and chemicals is easily produced chemical sproof the influence.So give full play to body's immunological function, in the novel influenza A H1N1 influenza virus of treatment, important effect arranged.
A kind of short corynebacteria preparation of the present invention or acellular short corynebacteria preparation are when being used for the novel influenza A H1N1 influenza virus medicament of preparation treatment, its usage is: 1-7 time/week, each 0.5～4mg, intramuscular injection or other suitable route administrations, 1～4 week was a course of treatment or followed the doctor's advice.Can be at interval with or use continuously several courses of treatment.
Because the easy variability of influenza virus, a lot of influenza virus have produced the drug resistance of enantiopathy cytotoxic drug, have limited the effectiveness of chemoprophylaxis and medicine, make the chemicals curative effect of a lot of resisiting influenza virus reduce or inefficacy.Current research is found novel influenza A H1N1 influenza virus to amantadine and rimantadine drug resistance, to Oseltamivir, zanamivir and oseltamivir phosphate capsule sensitivity, but curative effect and uncertain.Short corynebacteria preparation of the present invention or acellular short corynebacteria preparation confirm that through zoopery the infection of novel influenza A H1N1 influenza virus is had good therapeutical effect.
Used CPP, the NCP of the present invention is a kind of immunomodulator, is used for the novel influenza A H1N1 influenza virus medicament of preparation treatment, has following advantage:
1. the invention provides the new medical usage of CPP, NCP, provide new approach treating novel influenza A H1N1 influenza virus.
2. NCP preparation of the present invention has low pyrogen, no living contaminants, formaldehydeless residual, and granule has nanoscale, homogeneous grain diameter, and trap and diffusance improve, and help absorption of human body and improve drug effect; Simultaneously can reduce fever, chest pain, injection site redness, scleroma, side effect such as Liver and kidney toxicity are a kind of promising immune formulations, can be used as to be used for the novel influenza A H1N1 influenza virus medicament of preparation treatment.
3. preparation technology of the present invention is simple, can be made into oral liquid, injection, tablets and other formulations, and is easy to use.
4. NCP preparation of the present invention has overcome the novel influenza A H1N1 influenza virus of chemotherapy and has easily produced chemical sproof shortcoming.
The specific embodiment of preparation method of the present invention, absorbent properties and animal acute toxicity experiment is with patent ZL03157454.8.
The specific embodiment
The following specific embodiment is used to describe in detail the present invention, the present invention is not construed as limiting.
Experimental example 1NCP is to the protective effect of the novel H 1 N 1 influenza A virus infection of mice:
1. material and method
1.1, test sample: NCP (2mg/ml), deposit normal saline preparation during use for 4 ℃.0.25mg/ only, 0.5mg/, 1mg/ only
1.2, virus: novel influenza A H1N1 influenza virus, from WHO, in chick embryo allantoic cavity, go down to posterity, the experiment virus be E22～23 generations, the blood clotting titre is in (10～11 days embryos of Embryo Gallus domesticus age) more than 1/180.
1.3, mice: Kunming mouse, body weight 10～12 gram,
1.4, the protection test of mice influenza virus toxin: the novel influenza A H1N1 influenza virus of different time intravenous injection behind the mouse muscle injection NCP, establish the virus control group, observe deadly, calculate protective rate.
2. experimentation
2.1 animal model: the novel influenza A H1N1 influenza virus 1 * 10 of mouse mainline
5Virus/0.2ml/ only
2.2 dosage, grouping and administration
1) normal control group: 20, intramuscular injection normal saline 0.2ml/ only
2) virus control group: 20
3) high dose group: 1mg/0.2ml/, 20
4) middle dosage group: 0.5mg/0.2ml/, 20
5) low dose group: 0.25mg/0.2ml/, 20
NCP medication: 4 hours administered intramuscular behind the animal contaminated
Measure 2.3 observe
Observe all dead mouse situations, calculate protective rate.
Table 1: mouse muscle injection NCP after 24 hours to infecting the protective effect of novel influenza A H1N1 influenza virus
| Grouping | Death toll/infection number | Dead % | Protection % |
| High dose group | 1/20 | 5 | 95* |
| Middle dosage group | 0/20 | 0 | 100* |
| Low dose group | 2/20 | 10 | 90* |
| The virus control group | 16/20 | 80 | 20 |
| The normal control group | 0/0 | 0 |
*P＜0.01
As seen from Table 1, the NCP group has significant protective effect to the novel influenza A H1N1 influenza virus of 24 hours postoperative infections.Middle dosage group can protect 100%, makes mice avoid death, P＜0.01.The result shows that intramuscular injection NCP can obviously protect the infection of mice to novel influenza A H1N1 influenza virus.
Embodiment 2
1. material and method
1.1, test sample: CPP,
1.2, virus: novel influenza A H1N1 influenza virus, from WHO, in chick embryo allantoic cavity, go down to posterity, the experiment virus be E22～23 generations, the blood clotting titre is in (10～11 days embryos of Embryo Gallus domesticus age) more than 1/180.
1.3, mice: Kunming mouse, body weight 10～12 gram,
1.4, the protection test of mice influenza virus toxin: the novel influenza A H1N1 influenza virus of different time intravenous injection behind the mouse muscle injection CPP, establish the virus control group, observe deadly, calculate protective rate.
2. experimentation
2.1 animal model: the novel influenza A H1N1 influenza virus 1 * 10 of mouse mainline
5Virus/0.2ml/ only
2.2 dosage, grouping and administration
1) normal control group: 20, intramuscular injection normal saline 0.2ml/ only
2) virus control group: 20
3) high dose group: 6,000,000,000 cells/only, 20
4) dosage group in: 3,000,000,000 cells/only, 20
5) low dose group: 1,500,000,000 cells/only, 20
NCP medication: 4 hours administered intramuscular behind the animal contaminated
Measure 2.3 observe
Observe all dead mouse situations, calculate protective rate.
3. result
Table 2: mouse muscle injection CPP after 24 hours to infecting the protective effect of novel influenza A H1N1 influenza virus
| Grouping | Death toll/infection number | Dead % | Protection % |
| High dose group | 10/20 | 50 | 50* |
| Middle dosage group | 8/20 | 40 | 60* |
| Low dose group | 14/20 | 70 | 30* |
| The virus control group | 16/20 | 80 | 20 |
| The normal control group | 0/0 | 0 |
*P＜0.05
As seen from Table 2, the CPP group has the certain protection effect to the novel influenza A H1N1 influenza virus of 24 hours postoperative infections.Middle dosage group can protect 60%, makes mice avoid death, P＜0.05.The result shows, the protection mice that intramuscular injection CPP can be to a certain degree is to the infection of novel influenza A H1N1 influenza virus.
Claims (10)
1. short corynebacteria preparation or the acellular short corynebacteria preparation application in the novel H 1 N 1 influenza A virus infection medicament of preparation treatment.
2. the application in the novel H 1 N 1 influenza A virus infection medicament of preparation treatment of short corynebacteria preparation according to claim 1 or acellular short corynebacteria preparation, it is characterized in that wherein said short corynebacteria preparation is the preparation of being made by short corynebacteria behind formalin-inactivated.
3. the application in the novel H 1 N 1 influenza A virus infection medicament of preparation treatment of short corynebacteria preparation according to claim 1 or acellular short corynebacteria preparation, it is characterized in that, wherein said acellular short corynebacteria preparation is that granularity is less than 100nm by the cell-free preparation of short corynebacteria through the fragmentation preparation.
4. according to claim 1 or 3 described short corynebacteria preparations or the application of acellular short corynebacteria preparation in the novel H 1 N 1 influenza A virus infection medicament of preparation treatment, it is characterized in that, wherein said acellular short corynebacteria preparation pyrogen is less than 160EU/ml, protein content 10～40% (g/g), nucleic acid content 3～25% (g/g), all the other are polysaccharide.
5. according to claim 1 or 3 described short corynebacteria preparations or the application of acellular short corynebacteria preparation in the novel H 1 N 1 influenza A virus infection medicament of preparation treatment, it is characterized in that the granularity of wherein said acellular short corynebacteria preparation is 50～80nm.
6. the application of acellular short corynebacteria preparation according to claim 3 in the novel H 1 N 1 influenza A virus infection medicament of preparation treatment is characterized in that wherein said short corynebacteria is a propionibacterium acne, and bacterium number is 76-27, H-84 or 77-1.
7. the application in the novel H 1 N 1 influenza A virus infection medicament of preparation treatment according to claim 1 or 4 described acellular short corynebacteria preparations is characterized in that wherein said acellular short corynebacteria preparation pyrogen is less than 60EU/ml.Protein content 20～30% (g/g), nucleic acid content 3～10% (g/g), all the other are polysaccharide.
8. the application in the novel H 1 N 1 influenza A virus infection medicament of preparation treatment of short corynebacteria preparation according to claim 1 or acellular short corynebacteria preparation, it is characterized in that wherein said acellular short corynebacteria preparation prepares by following method:
1) short corynebacteria work seed lot strain is continuous is inoculated in big bottle or nutrition jar after going down to posterity for 2～4 times and supports in the base and cultivated 5～7 days:
2) thalline of the no living contaminants of collection, heated and boiled obtained bacterium liquid in 15～60 minutes;
3) the bacterium liquid that sterility test is qualified is used the broken thalline of breaker after washing:
4) collecting precipitation after centrifugal of the suspension behind the bacterial cell disruption, suspension is made in washing precipitation:
5) with the sterilization of suspension packing post-heating, obtain acellular short corynebacteria preparation.
9. the application in the novel H 1 N 1 influenza A virus infection medicament of preparation treatment of short corynebacteria preparation according to claim 8 or acellular short corynebacteria preparation is characterized in that wherein said condition of culture is 36～37 ℃; Described culture medium is not for containing the sulphur glycollate culture medium of agar; Described thalline is an inactivated bacteria, described breaker is that bacterial cell disruption is arrived less than 100nm, preferred size is the superhigh pressure water jet collider of 50～80nm, broken under aseptic condition, carry out: the washing described in the step 3) be with precipitate with physiological saline solution centrifuge washing at least 2 times: the washing described in the step 4) be with precipitate with physiological saline solution centrifuge washing 0-5 time: the centrifugal condition of suspension is under 4 ℃～room temperature behind the bacterial cell disruption, centrifugal 30～150 minutes of 6000～12000rpm; Being heated to be described in the step 5) 60～65 ℃ of heating 0.5～2 hour.
10. the application in the novel H 1 N 1 influenza A virus infection medicament of preparation treatment of short corynebacteria preparation according to claim 8 or acellular short corynebacteria preparation, it is characterized in that, with the acellular short corynebacteria preparation lyophilization that step 5) obtains, obtain the lyophilized formulations of acellular short corynebacteria.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011071134A1 (en) * | 2009-12-10 | 2011-06-16 | 明治乳業株式会社 | Prophylactic composition for influenza infection |
| CN115607508A (en) * | 2022-09-09 | 2023-01-17 | 杭州医学院 | Nasal administration preparation for preventing and treating pneumonia caused by influenza A virus, and preparation method and application thereof |
| WO2024079492A1 (en) * | 2022-10-12 | 2024-04-18 | Semmelweis Egyetem | Corynebacterium strains, combinations, and lyophilized formulations thereof for use in the prevention of a viral infection |
-
2009
- 2009-06-12 CN CN2009100865937A patent/CN101919878A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011071134A1 (en) * | 2009-12-10 | 2011-06-16 | 明治乳業株式会社 | Prophylactic composition for influenza infection |
| CN102665738A (en) * | 2009-12-10 | 2012-09-12 | 株式会社明治 | Prophylactic composition for influenza infection |
| CN115607508A (en) * | 2022-09-09 | 2023-01-17 | 杭州医学院 | Nasal administration preparation for preventing and treating pneumonia caused by influenza A virus, and preparation method and application thereof |
| CN115607508B (en) * | 2022-09-09 | 2024-03-15 | 杭州医学院 | Nasal administration preparation for preventing and treating pneumonia caused by influenza A virus and preparation method and application thereof |
| WO2024079492A1 (en) * | 2022-10-12 | 2024-04-18 | Semmelweis Egyetem | Corynebacterium strains, combinations, and lyophilized formulations thereof for use in the prevention of a viral infection |
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