CN101874896A - Immune potentiator and recombinant IFN-alpha combined medicament - Google Patents
Immune potentiator and recombinant IFN-alpha combined medicament Download PDFInfo
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- CN101874896A CN101874896A CN2009100686975A CN200910068697A CN101874896A CN 101874896 A CN101874896 A CN 101874896A CN 2009100686975 A CN2009100686975 A CN 2009100686975A CN 200910068697 A CN200910068697 A CN 200910068697A CN 101874896 A CN101874896 A CN 101874896A
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Abstract
The invention relates to an immune potentiator and recombinant IFN-alpha combined medicament. In the recombinant IFN-alpha and a compound preparation having synergism with the recombinant IFN-alpha, an immune potentiator may be one or the combination of several of vitamin C, poly IC, lactobacillus and pig spleen transfer factors. While treating diseases, the medicament can stimulate the organisms to continuously generate interferon so as to form a continuous treating process and achieve an aim of thoroughly curing diseases; and in addition, the medicament can also strengthen the resisting power of the organisms and have the effects of preventing diseases and avoiding secondary infection of diseases.
Description
Technical field
The invention belongs to medical technical field, relate to a kind of recombinant IFN-alpha and recombinant IFN-alpha is had the composite reagent thing of potentiation.
Background technology
Porcine epizootic diarrhea (PED) and transmissible gastroenteritis of swine (TGE) are the viral infectious of two kinds of serious threat pig industrys, its cause of disease Porcine epidemic diarrhea virus (PEDV) and transmissible gastro-enteritis virus (TGEV) belong to coronavirus, plesiomorphism, the target organ of breeding is identical.These two kinds of sick outbreak of epidemic usually cause large quantities of death of newborn piglet.Existing general solution is vaccinated exactly, yet for various reasons, can cause immuning failure unavoidably sometimes, and the outburst disease brings certain difficulty for diagnosis and treatment.Behind immuning failure, often rely on antibiotic therapy, caused the increase of Resistant strain and the generation of cross infection.Therefore find a kind of better treatment preventive drug very important.
Interferon (English abbreviation: IFN) be a kind of widely used clinically biologics with broad-spectrum antiviral, antitumor and immunoloregulation function, be divided into α, β and γ three classes by its antigenicity difference, and alpha-interferon has the hypotype more than 20, as Intederon Alpha-2a, Alfacon-1 b and Interferon Alpha-2b etc., beta-interferon has 4 hypotypes, and gamma interferon has 1 hypotype.
Pig interferon is to be studied one of animal interferon the earliest, and the relevant report of pig interferon is just arranged the eighties in 20th century.In recent years, the inductive condition and the physics and chemistry activity of pig interferon there has been further research, simultaneously leukocyte interferon of pig carried out a large amount of clinical trials.Studies show that, it is to many viral infectious of pig, as epidemic diarrhea, swine fever, transmissible gastroenteritis etc., and to bovine viral diarrhoea, gosling plague, lamb diarrhoea waits all has good curative effect, and test confirms to exist between the animals such as pig interferon and cattle and sheep and intersects activity.Recent years, the molecular cloning of pig interferon α, β, γ gene and sequence analysis are succeedd.Thank to the IFN-α gene that employing pcr clones such as Potiria pectinifera (Mukller et Tro Sehel) obtain and form 166 aminoacid of encoding altogether by 501 nucleotide; The porcine beta interferon IFN-beta gene fragment that clones such as Xia Chun obtain has 668 nucleotide, 186 aminoacid of encoding.Cao Ruibing etc. are from amplifying pig IFN-γ gene through the pig peripheral blood leucocyte of ConA inducing culture, after transforming, insert prokaryotic expression carrier PRLG, and realized efficiently expressing in escherichia coli, expression product exists with the inclusion body form, handle through degeneration, renaturation, desalination, gel chromatography, reorganization pig IFN-γ has higher interferon activity.Simultaneously; Wan Jianqing, Chen Tao etc. successfully give expression to the recombinant interferon gene respectively in pichia yeast expression system and escherichia expression system; expression product accounts for the ratio of bacterial protein between 20%～35%; the antiviral activity that expression product has, for the large-scale production of genetic engineering interferon and application provide may.
Yet, in with interferon therapy lysis, can stimulate body produce interferon antibody (anti--IFN), make therapeutic effect descend or state of an illness bounce-back, there is clinical research to show, the use interferon of long period can make probability that body produces interferon antibody more than 40%, makes that cure rate and effective percentage are not that the treatment rate of very high interferon significantly reduces.In order to address the above problem, the invention provides a kind of compound preparation of recombinant IFN-alpha, treat than independent use porcine alpha-interferon and have better therapeutic effect.
Summary of the invention
The compound preparation that the purpose of this invention is to provide a kind of recombinant IFN-alpha.
A kind of compound preparation of recombinant IFN-alpha is characterized in that: its main component is porcine alpha-interferon and immunostimulant.
A kind of compound preparation of recombinant IFN-alpha is characterized in that: immunostimulant is selected one or more in vitamin C, polyinosini, lactobacillus and the transfer factor of pig usefulness for use.
A kind of compound preparation of recombinant IFN-alpha is characterized in that their combination matching is: the vitamin C combination of per 1,000,000 IU recombinant IFN-alphas and 200～500 milligrams; The polyinosini combination of per 1,000,000 IU recombinant IFN-alphas and 0.01～1 milligram; The lactobacillus combination of per 1,000,000 IU recombinant IFN-alphas and 40～100 milligrams; The Lien Sus domestica transfer factor combination of per 1,000,000 IU recombinant IFN-alphas and 40～100 milligrams;
Wherein be preferable over: the vitamin C combination of per 1,000,000 IU recombinant IFN-alphas and 200 milligrams; The polyinosini combination of per 1,000,000 IU recombinant IFN-alphas and 1 milligram; The lactobacillus combination of per 1,000,000 IU recombinant IFN-alphas and 40 milligrams; The Lien Sus domestica transfer factor combination of per 1,000,000 IU recombinant IFN-alphas and 40 milligrams.
Vitamin C can improve leukocyte and phagocytic function, promotes reticuloendothelial system and antibody to form, and strengthens the ability of anti-stress, safeguards liver detoxification, improves cardiovascular function.
Polyinosini is called polyinosinic acid, Poly I:C again, combine by polymolecular nucleotide, in vivo can inducement interferon, it is a kind of efficient interferon inducers, the disease that multiple virus is caused has curative effect preferably, and can strengthen antibody formation and stimulate the macrophage phagocytic effect.
The lactobacillus goods can produce the nonspecific immunity regulatory factor, improve the immunity of animal.The lactobacillus goods of directly feeding can improve the antibody horizontal of body, produce interferon, improve immunoglobulin concentration and huge cytophilic activity, enhancing human body immunity function and premunition.
The Lien Sus domestica transfer factor is a class low molecular peptide and a nucleotide complex that extracts from Lien Sus domestica, have the immunologic cellular activity of exciting, regulate effects such as immunologic function, enhancing body non-specific cell immunologic function, be described as the triggering agent of T cytoactive, the reinforcing agent of cellular immunization, cell immunomodulator and interferon produce and start agent.The transfer factor molecular weight is little, nontoxic, no antigen, do not play anaphylaxis, and antibody and can to surmount kind be advantage such as boundary application does not create antagonism.Since the famous immunology expert Lawrence of the U.S. found and studies transfer factor, many theoretical researches had not only been carried out in countries in the world, and have carried out a large amount of clinical application researchs, make transfer factor become present widely used enhance immunity preparation.
First purpose of the present invention provides a kind of compound preparation of recombinant IFN-alpha, it is characterized in that being made up of recombinant IFN-alpha and immunostimulant.Wherein immunostimulant is selected one or more in vitamin C, polyinosini, lactobacillus and the transfer factor of pig usefulness for use.
Second purpose of the present invention is that medicine of the present invention can make various dosage forms as required, and in specific embodiments, wherein said dosage form comprises dosage forms such as spray, injection, lyophilized powder, oral powder.
Be that example is carried out following explanation with recombinant IFN-alpha and immunostimulant injection below:
1. recombinant IFN-alpha and ascorbic combination aqueous injection
Prescription:
Recombinant IFN-alpha 3 * 10
4Ten thousand IU
Mannitol 15ml
Vitamin C 60.0g
The about 7.2g of sodium bicarbonate (transferring pH5.0-7.0)
EDTA 0.015g
Sodium pyrosulfite 0.6g
Water for injection adds to 300ml
Recombinant IFN-alpha and ascorbic combination aqueous injection technological process:
In recombinant IFN-alpha provided by the invention and the ascorbic composite reagent, every milliliter of vitamin C that contains 1,000,000 recombinant IFN-alpha IU and 200 milligrams.
2. the combination injection of recombinant IFN-alpha and polyinosini
The preparation technology of polyinosini injection:
(1) sodium chloride-phosphate buffer accurately takes by weighing NaH
2PO
42H
2O:0.262g, Na
2HPO
412H
2O:1.546g, NaCl:8.768g adds an amount of water for injection and makes dissolving, and is settled to 1000ml with water for injection.Get about 500ml, 45 ℃ are incubated 10 minutes;
(2) accurately take by weighing polyinosinic acid 0.5g, poly 0.5g adds the sodium chloride of good heat insulation-phosphate buffer 60ml respectively, stirs to make dissolving.Sealing was in 45 ℃ of insulations 20 minutes;
(3) polyinosinic acid, the poly solution with above-mentioned good heat insulation mixes, stir, and sealing, 45 ℃ are incubated 10 minutes;
(4) mixed solution of above-mentioned good heat insulation adds sodium chloride-phosphate buffered solution and is settled to 1000ml, fill, and sterilization, promptly;
Prescription:
Recombinant IFN-alpha 1 * 10
5Ten thousand IU
Mannitol 50ml
Polyinosini injection 1000ml
Glycerol 5%
Chloroform 0.3%
Zinc stearate 1%
NaHCO
3Buffer agent an amount of (transferring pH6.0-7.0)
The combination of recombinant IFN-alpha and polyinosini aqueous injection technological process:
In the composite reagent of recombinant IFN-alpha provided by the invention and polyinosini, every milliliter of polyinosini that contains 1,000,000 recombinant IFN-alpha IU and 1 milligram.
3. the combination injection of recombinant IFN-alpha and lactobacillus
Prescription:
Recombinant IFN-alpha 1 * 10
4Ten thousand IU
Lactobacillus 4.0g
Mannitol 5ml
Glycerol 5%
Chloroform 0.3%
Zinc stearate 1%
NaHCO
3Buffer agent an amount of (transferring pH5.0-6.5)
Water for injection adds to 100ml
The combination of recombinant IFN-alpha and lactobacillus injection technological process:
In the composite reagent of recombinant IFN-alpha provided by the invention and lactobacillus, the combination of the lactobacillus of per 1,000,000 recombinant IFN-alpha IU and 40 milligrams.
4. the combination aqueous injection of recombinant IFN-alpha and Lien Sus domestica transfer factor
The preparation technology of Lien Sus domestica transfer factor:
(1) with refrigerated pig spleen 1kg, remove fat, tunicle and big blood vessel, spend and be cut into small pieces after the pyrogen distilled water is cleaned, add the 1L physiological saline solution;
(2) carry out cell breakage with the even system machine of high pressure, 1% chitosan solution that adds 200mL is simultaneously made homogenate as flocculating agent behind the mixing;
(3) put-20 ℃ of freezing and room temperature thawings, multigelation 10 times;
(4) then at 4 ℃, centrifugal and get supernatant with 10000rpm, after 3 times, supernatant is placed bag filter and bag filter is put into the container that fills physiological saline solution repeatedly, place 0-4 ℃ of dialysis 48h;
(5) use 10%NaHCO
3Adjust the dialysis solution pH value to neutral, and add 0.3% chloroform give antiseptic;
(6) dialysis solution is filtered by microporous filter membrane, carry out ultrafiltration with ultrafilter membrane then, be the stock solution that contains transfer factor;
(7) detecting stock solution Lien Sus domestica transfer factor content is 40mg/mL.
Prescription:
Recombinant IFN-alpha 1 * 10
5Ten thousand IU
Lien Sus domestica transfer factor solution 1000mL
Mannitol 50ml
Glycerol 5%
Chloroform 0.3%
Zinc stearate 1%
NaHCO
3Buffer agent an amount of (transferring pH6.0～7.0)
The combination of recombinant IFN-alpha and Lien Sus domestica transfer factor aqueous injection technological process:
In the composite reagent of recombinant IFN-alpha provided by the invention and Lien Sus domestica transfer factor, every milliliter of Lien Sus domestica transfer factor that contains 1,000,000 recombinant IFN-alpha IU and 40 milligrams.
Be example with recombinant IFN-alpha and Lien Sus domestica transfer factor combination with aqueous injection below, use description of test medicine of the present invention therapeutic effect clinically:
The veterinary biologics clinical experimental study data requirement of announcing No. 442 issue according to the Ministry of Agriculture carries out following research:
1, experimental animal: 28 of the healthy piglets of 3 ages in days, be divided into 4 groups, 7 every group, blank group, medication group of the present invention, single with recombinant IFN-alpha medicine matched group, the positive group of the not administration of falling ill.Neutralization test turns out to be the transmissible gastroenteritis of swine negative antibody, and usual manner is fed.See the following form
Table 1 test grouping and processing
2, counteracting toxic substances: 21 piglets are attacked 100TCID
50Transmissible gastro-enteritis virus all causes morbidity; Blank is organized not counteracting toxic substances;
3, test method: treat that symptom occurs, carry out the clinical treatment test immediately.Medication group intramuscular injection transfer factor of the present invention and recombinant IFN-alpha compound preparation, the 0.5ml/ head; Medicine matched group intramuscular injection recombinant IFN-alpha 500,000 units/head; Once a day, logotype is 3.
4, result of the test: medicine matched group symptom behind logotype medicine on the 3rd alleviates slightly, lassitude, and appetite reduces, and more the back is bad; Sick pig mental restoration is good after the medication group of the present invention medication, and it is normal that appetite is recovered, favourable prognosis.
Beneficial effect of the present invention: the object of the present invention is to provide a kind of compound medicine by recombinant IFN-alpha and immunostimulant combination.The present invention adopts one or several immunostimulants as recombinant IFN-alpha in vitamin C, polyinosini, lactobacillus and the transfer factor of pig usefulness, and adds suitable pharmaceutic adjuvant and make compound preparation through certain technology.Adopt medicine of the present invention to treat, the immunostimulant when using the interferon therapy disease in the medicine can induce body to produce interferon, thereby forms the therapeutic process that continues, and reaches the purpose of thorough cure diseases.Interact between the medicine, impel the body persistence to produce interferon, effectively treat disease, the resistance of enhancing body plays prophylactic generation and prevents the effect of secondary infection, is suitable for applying in animal husbandry.
The specific embodiment
Embodiment one: recombinant IFN-alpha and ascorbic combination aqueous injection
Get recombinant IFN-alpha 3 * 10
5Ten thousand IU, vitamin C 60.0 grams, pharmaceutic adjuvant selects for use EDTA, sodium pyrosulfite and sodium bicarbonate buffer agent an amount of, add the suitable quantity of water dissolving, regulate pH value between 5.0～7.0, filter sterilization, be distributed into 10 bottles, every bottle of 30mL seals and promptly makes every bottle of compound recipe aqueous injection that contains 200 milligrams of recombinant IFN-alpha 1,000,000 IU and vitamin Cs.
Use: be mainly used in treatment transmissible gastroenteritis of swine disease, Porcine Epidemic Diarrhea.
Usage and consumption: intramuscular injection, amount once, piglets (30kg following) this product 0.5mL, in (30-60kg) this product 1.0mL, big pig (more than the 60kg) this product 1.5mL, every day 1 time, logotype 2-3 days.
Embodiment two: the combination aqueous injection of recombinant IFN-alpha and polyinosini
Get recombinant IFN-alpha 1 * 10
5Ten thousand IU, polyinosini injection 1000ml (polyinosini content 1mg/ml), pharmaceutic adjuvant selects for use chloroform, zinc stearate and sodium carbonate salt buffer agent an amount of, regulate pH value between 6.0～7.0, filter sterilization, be distributed into 100 bottles, every bottle of 10mL seals and promptly makes every bottle of compound recipe aqueous injection that contains 1 milligram of recombinant IFN-alpha 1,000,000 IU and polyinosini.
Use: broad-spectrum antiviral drug.Mainly be applicable to the various zoosis toxicity diseases of treatment, the especially treatment of the viral disease of bringing out after the fowl poultry immune failure is as swine fever, SVD, foot and mouth disease, transmissible gastroenteritis of swine etc. and immunodeficiency or hypoimmunity disease.This product also has good killing action to some antibacterial, rickettsia, chlamydia and some protozoon of infecting in the host cell.
Usage and consumption: intramuscular injection, once measure, every 1kg body weight, this product 0.02ml, every injection on the 3rd once.
Embodiment three: the combination aqueous injection of recombinant IFN-alpha and lactobacillus
Get recombinant IFN-alpha 1 * 10
4Ten thousand IU, lactobacillus 4 grams, pharmaceutic adjuvant selects for use glycerol, chloroform, zinc stearate and phosphate buffer an amount of, add the injection water dissolving, regulate pH value between 5.0～6.5, filter sterilization, be distributed into 100 bottles, every bottle of 1mL seals and promptly makes every bottle of compound recipe aqueous injection that contains 40 milligrams of recombinant IFN-alpha 1,000,000 IU and lactobacilluss.
Use: be used for the treatment of yellow scour of piglet, Hakuri, diarrhea of weaned piglets, transmissible gastroenteritis of swine disease, Porcine Epidemic Diarrhea.
Usage and consumption: intramuscular injection, amount once, piglets (30kg following) this product 0.5mL, in (30-60kg) this product 1.0mL, big pig (more than the 60kg) this product 1.5mL, every day 1 time, logotype 2-3 days.
Embodiment four: recombinant IFN-alpha and pig are with the combination aqueous injection of transfer factor
Get recombinant IFN-alpha 1 * 10
4Ten thousand IU, Lien Sus domestica transfer factor solution 1000ml (Lien Sus domestica transfer factor content 40mg/ml), pharmaceutic adjuvant selects for use chloroform, zinc stearate, glycerol and carbonate buffer agent an amount of, regulate pH value between 6.0～7.0, filter sterilization, be distributed into 100 bottles, every bottle of 10mL seals and promptly makes every bottle of compound recipe aqueous injection that contains recombinant IFN-alpha 1,000,000 IU and pig with 40 milligrams of transfer factors.
Use: each viroid disease of treatment pig, as diseases such as swine fever, transmissible gastroenteritis of swine disease, Porcine Epidemic Diarrhea.
Usage and consumption: intramuscular injection, amount once, piglets (30kg following) this product 0.5mL, in (30-60kg) this product 1.0mL, big pig (more than the 60kg) this product 1.5mL, every day 1 time, logotype 2-3 days.
Claims (4)
1. the compound preparation of a recombinant IFN-alpha is characterized in that being grouped into by following one-tenth: recombinant IFN-alpha and immunostimulant.
2. the compound preparation of a kind of recombinant IFN-alpha according to claim 1 and immunostimulant is characterized in that immunostimulant comprises one or more combinations in vitamin C, polyinosini, lactobacillus and the Lien Sus domestica transfer factor.
3. the composite reagent thing that comprises recombinant IFN-alpha and its immunostimulant according to claim 1 and 2 is characterized in that their combination matching is: per 1,000,000 IU recombinant IFN-alphas make up with 200～500 milligrams vitamin C; The polyinosini combination of per 1,000,000 IU recombinant IFN-alphas and 0.01～1 milligram; The lactobacillus combination of per 1,000,000 IU recombinant IFN-alphas and 40～100 milligrams; The Lien Sus domestica transfer factor combination of per 1,000,000 IU recombinant IFN-alphas and 40～100 milligrams;
Wherein be preferable over: the vitamin C combination of per 1,000,000 IU recombinant IFN-alphas and 200 milligrams; The polyinosini combination of per 1,000,000 IU recombinant IFN-alphas and 1 milligram; The lactobacillus combination of per 1,000,000 IU recombinant IFN-alphas and 40 milligrams; The Lien Sus domestica transfer factor combination of per 1,000,000 IU recombinant IFN-alphas and 40 milligrams.
4. according to the compound preparation of claim 1,2 or 3 described a kind of recombinant IFN-alphas and immunostimulant, it is characterized in that to make dosage forms such as spray, injection, lyophilized powder, oral powder as required.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104208674A (en) * | 2013-09-30 | 2014-12-17 | 郑州后羿制药有限公司 | Mixed freeze-dried powder for preventing pig viral diseases and preparation method thereof |
| CN104208679A (en) * | 2013-09-30 | 2014-12-17 | 郑州后羿制药有限公司 | Composition for preventing porcine circovirus disease, mixed freeze-dried powder, and preparation method of the composition |
| CN104686803A (en) * | 2015-02-11 | 2015-06-10 | 岳道友 | Compound micro-ecological preparation for promoting growth of animals and improving production performance of animals |
| WO2022087471A1 (en) * | 2020-10-22 | 2022-04-28 | Health Research, Inc. | Synergistic induction of immunity against rna viruses |
-
2009
- 2009-04-29 CN CN2009100686975A patent/CN101874896A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104208674A (en) * | 2013-09-30 | 2014-12-17 | 郑州后羿制药有限公司 | Mixed freeze-dried powder for preventing pig viral diseases and preparation method thereof |
| CN104208679A (en) * | 2013-09-30 | 2014-12-17 | 郑州后羿制药有限公司 | Composition for preventing porcine circovirus disease, mixed freeze-dried powder, and preparation method of the composition |
| CN104208674B (en) * | 2013-09-30 | 2016-08-17 | 郑州后羿制药有限公司 | A kind of freeze-dried mixed powder of anti-porcine viral diseases and preparation method thereof |
| CN104208679B (en) * | 2013-09-30 | 2016-08-17 | 郑州后羿制药有限公司 | Compositions, freeze-dried mixed powder and the preparation method that a kind of resisting porcine circovirus is sick |
| CN104686803A (en) * | 2015-02-11 | 2015-06-10 | 岳道友 | Compound micro-ecological preparation for promoting growth of animals and improving production performance of animals |
| WO2022087471A1 (en) * | 2020-10-22 | 2022-04-28 | Health Research, Inc. | Synergistic induction of immunity against rna viruses |
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Application publication date: 20101103 |