CN103877566A - Swine broad-spectrum immune enhancement preparation and preparation method thereof - Google Patents
Swine broad-spectrum immune enhancement preparation and preparation method thereof Download PDFInfo
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- CN103877566A CN103877566A CN201410062985.0A CN201410062985A CN103877566A CN 103877566 A CN103877566 A CN 103877566A CN 201410062985 A CN201410062985 A CN 201410062985A CN 103877566 A CN103877566 A CN 103877566A
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Abstract
The invention relates to the field of veterinary biological pharmacy, and discloses a swine broad-spectrum immune enhancement preparation and a preparation method thereof. The main components of the swine broad-spectrum immune enhancement preparation comprise thymosin-interferon fusion protein and chitosan. The preparation method of the swine broad-spectrum immune enhancement preparation comprises the following basic preparation processes: (1) dissolving chitosan in an acetic acid-sodium acetate buffer solution; (2) dissolving the thymosin-interferon fusion protein in a sodium polyphosphate solution; (3) dropwise adding a solution obtained in the step (2) to a solution obtained in the step (1) under a magnetic stirring condition, and reacting at room temperature, thus obtaining the thymosin-interferon fusion protein-chitosan immune enhancement preparation. The immune enhancement preparation is simple in production technology, strong in production link controllability and beneficial to wide popularization; moreover, compared with a single preparation, the swine broad-spectrum immune enhancement preparation has the advantages that the actual amount of usage is obviously reduced, the effect of the broad-spectrum immune enhancement preparation is more superior to that of the single preparation, and the broad-spectrum immune enhancement preparation can play a positive role in prevention and control of swine diseases.
Description
Technical field
The present invention relates to field of biological pharmacy for animals, particularly relate to boar wide spectrum immunostimulant preparation and preparation method.
Background technology
Immunostimulant refers to the material that while use with antigen individually or simultaneously, energy enhancing human body immunity is replied, and can produce by non-specific approach enhancing body the ability of specific immune response to antigenic substance; The advantage that it is efficient, stable, nontoxic, medically has important value and effect.Because it can activate one or more immunocytes, non-specific and the specific immune function of enhancing body, make cellular immunization and the humoral immune function of hypofunction recover normal, reach the curative effect of enhancing human body immunity power, clinical treatment disease and immunodeficiency and the malignant tumor relevant with immunologic hypofunction of being mainly used in, also for controlling antibacterial, virus and fungal infection.Conventional immunostimulant kind is a lot of clinically, can be divided into immunological adjuvant, biological immunopotentiator and feeding immunostimulant etc. by effect, and wherein, interferon and thymosin are to apply biological immunopotentiator more widely.
Interferon (IFN) is the little skin of emiocytosis, has antiviral, the biologic activity widely such as anti-hypertrophy and immunomodulating.Thymosin (THY) is many skins of the class hormone of organizing epithelial cells by thymus, there is immunoregulation effect, strengthen NK cell function, promote CD3+, CD4+, CD8+ hyperplasia, the apoptosis that anti-CD3+ brings out, suppresses a series of biological functions such as tumor growth.Evidence; IFN and THY are prepared into IFN-THY fusion rotein; not only having certain Cell protection avoids the activity of virus attack and promotes the activity that cell surface receptor is expressed; kept preferably IFN and and THY biological function separately, and the dead application of fusion rotein can significantly improve IFN and the antiviral effect of THY.
Chitosan (Chitosna, CS) is the de-second phthalein product of chitin, is also desacetoxy chitin, relative molecular mass from hundreds thousand of to millions of not etc., water insoluble and aqueous slkali.CS, as the natural polycation marine polysaccharide of one, has the free radical such as amino, light base in molecule, show physics, the chemistry and biology characteristic of many uniquenesses.It has good biological degradability and biocompatibility, can form composite membrane with many kinds of substance, aspect medicine control release vehicle, artificial skin and other biomaterial oneself wooden dipper widely application.For CS weak solution, on the one hand, the ionic strength in state and the solution of CS in weak solution has much relations, in addition, the form that the viscosity of CS weak solution and CS molecule exist in solution also has very large relation, and the ionic strength of solution directly has influence on again the state of CS molecule in solution.At present medicine carrying CS nanoparticle has become the study hotspot in pharmaceutics field, and CS nanoparticle can slow releasing pharmaceutical, and target administration also can increase the stability of the medicine such as albumen, nucleotide, makes its packaging medicine be difficult for being degraded.
At present, using CS as carrier, the pig immunostimulant that coated IFN-THY fusion rotein is prepared into is not also applied to veterinary clinic.This research invention will rely on biotechnology, on the basis of efficient preparation IFN-THY fusion rotein, utilize CS as carrier, be prepared into pig wide spectrum immunostimulant preparation, realize industrialized large-scale production, for the sound development of animal husbandry provides sound assurance.
Summary of the invention
The object of the present invention is to provide boar wide spectrum immunostimulant preparation and preparation method.
For achieving the above object, a boar wide spectrum immunostimulant preparation and preparation method provided by the invention, main component comprises thymosin-interferon fusion protein and chitosan, the basic process of preparation is as follows:
(1) chitosan is dissolved in NaAc_HAc buffer solution;
(2) thymosin-interferon fusion protein is dissolved in polyphosphoric acids sodium solution;
(3), under magnetic agitation, the solution that (2) are obtained is added drop-wise in the solution that (1) obtain, and under room temperature, reacts.
Further, the pH value of NaAc_HAc buffer solution is 4.5 ~ 6.
Further, the quality percent by volume of chitosan and NaAc_HAc buffer solution is 0.5 ~ 5%.
The molecular weight of the chitosan of further, selecting is at 200 ~ 600 kDa.
Further, the quality percent by volume of thymosin-interferon fusion protein and polyphosphoric acids sodium solution is 1 ~ 5%.
Further, the acetic acid-sodium acetate solution of chitosan-containing is 3:1 ~ 10:1 with the volume ratio of the polyphosphoric acids sodium solution containing thymosin-interferon fusion protein.
Further, the acetic acid-sodium acetate solution of chitosan-containing is 2 ~ 5 hours with the time of at room temperature reacting of the polyphosphoric acids sodium solution containing thymosin-interferon fusion protein.
The invention has the beneficial effects as follows:
(1) pig of the present invention is simple with wide spectrum immunostimulant preparation production technique, and production link controllability is strong, is conducive to extensive popularization.
(2) the obvious minimizing of the more single preparation of actual use amount of this immunostimulant system, effect is more better than the effect of single preparation.
The specific embodiment
Below in conjunction with embodiment, the present invention will be further described in detail, and advantage and disadvantage of the present invention will be more clear along with description.But these embodiment are only exemplary, scope of the present invention are not formed to any restriction.It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can the details of technical solution of the present invention and form be modified or be replaced, but these modifications and replacement all fall into protection scope of the present invention.
Embodiment 1
1. the preparation of CS solution
Adopt ion gelling process to prepare chitosan (CS) nanoparticle, it is acetic acid-sodium acetate (HAC-NaAC) buffer (pH 4.5) that first dose volume mark is 1%, again the addition of C S is dissolved in HAC-NaAC buffer, the quality percent by volume that makes CS and HAC-NaAC buffer solution is 1%, spend the night fully swelling, thereby be mixed with CS solution.
2. the preparation of THY-IFN-TPP solution
Compound concentration is 2gL
-1sodium polyphosphate (TPP) solution, get appropriate thymosin-interferon (THY-IFN) fusion rotein and be dissolved in TPP solution, the quality percent by volume that makes THY-IFN fusion rotein and TPP solution is 1%, thereby is mixed with THY-IFN-TPP solution.
3. the preparation of CS-THY-IFN immunostimulant
Under room temperature magnetic agitation, the THY-IFN-TPP solution of preparation is slowly added drop-wise in CS solution, and the volume ratio of CS solution and THY-IFN-TPP solution is 3:1, at room temperature reacts 2 hours, obtain CS-THY-IFN-TPP solution, be CS-THY-IFN immunostimulant preparation.
Embodiment 2
In the present embodiment, prepare in the process of CS-THY-IFN immunostimulant, only change partial condition wherein, preparation procedure is identical with embodiment 1, specific as follows:
1. the preparation of CS solution
Dose volume mark is 1% HAC-NaAC buffer (pH 5), then the addition of C S is dissolved in HAC-NaAC buffer, and making the quality percent by volume of CS and HAC-NaAC buffer solution is 1.5%, spends the night fully swelling, thereby is mixed with CS solution.
2. the preparation of THY-IFN-TPP solution
Compound concentration is 2gL
-1tPP solution, get appropriate THY-IFN fusion rotein and be dissolved in TPP solution, the quality percent by volume that makes THY-IFN fusion rotein and TPP solution is 2%, thereby is mixed with THY-IFN-TPP solution.
3. the preparation of CS-THY-IFN immunostimulant
Under room temperature magnetic agitation, the THY-IFN-TPP solution of preparation is slowly added drop-wise in CS solution, and the volume ratio of CS solution and THY-IFN-TPP solution is 4:1, at room temperature reacts 2 hours, obtain CS-THY-IFN-TPP solution, be CS-THY-IFN immunostimulant preparation.
Embodiment 3
In the present embodiment, prepare in the process of CS-THY-IFN immunostimulant, only change partial condition wherein, preparation procedure is identical with embodiment 1, specific as follows:
1. the preparation of CS solution
Dose volume mark is 1% HAC-NaAC buffer (pH 5), then the addition of C S is dissolved in HAC-NaAC buffer, and making the quality percent by volume of CS and HAC-NaAC buffer solution is 2%, spends the night fully swelling, thereby is mixed with CS solution.
2. the preparation of THY-IFN-TPP solution
Compound concentration is 2gL
-1tPP solution, get appropriate THY-IFN fusion rotein and be dissolved in TPP solution, the quality percent by volume that makes THY-IFN fusion rotein and TPP solution is 3%, thereby is mixed with THY-IFN-TPP solution.
3. the preparation of CS-THY-IFN immunostimulant
Under room temperature magnetic agitation, the THY-IFN-TPP solution of preparation is slowly added drop-wise in CS solution, and the volume ratio of CS solution and THY-IFN-TPP solution is 5:1, at room temperature reacts 3 hours, obtain CS-THY-IFN-TPP solution, be CS-THY-IFN immunostimulant preparation.
Embodiment 4
In the present embodiment, prepare in the process of CS-THY-IFN immunostimulant, only change partial condition wherein, preparation procedure is identical with embodiment 1, specific as follows:
[0027]1. the preparation of CS solution
Dose volume mark is 1% HAC-NaAC buffer (pH 5.5), then the addition of C S is dissolved in HAC-NaAC buffer, and making the quality percent by volume of CS and HAC-NaAC buffer solution is 2%, spends the night fully swelling, thereby is mixed with CS solution.
2. the preparation of THY-IFN-TPP solution
Compound concentration is 2gL
-1tPP solution, get appropriate THY-IFN fusion rotein and be dissolved in TPP solution, the quality percent by volume that makes THY-IFN fusion rotein and TPP solution is 2.5%, thereby is mixed with THY-IFN-TPP solution.
3. the preparation of CS-THY-IFN immunostimulant
Under room temperature magnetic agitation, the THY-IFN-TPP solution of preparation is slowly added drop-wise in CS solution, and the volume ratio of CS solution and THY-IFN-TPP solution is 6:1, at room temperature reacts 4 hours, obtain CS-THY-IFN-TPP solution, be CS-THY-IFN immunostimulant preparation.
Claims (7)
1. wide spectrum immunostimulant preparation and preparation method for a boar, is characterized in that: main component comprises thymosin-interferon fusion protein and chitosan, and the basic process of preparation is as follows:
(1) chitosan is dissolved in NaAc_HAc buffer solution;
(2) thymosin-interferon fusion protein is dissolved in polyphosphoric acids sodium solution;
(3), under magnetic agitation, the solution that (2) are obtained is added drop-wise in the solution that (1) obtain, and under room temperature, reacts.
2. by wide spectrum immunostimulant preparation and preparation method for pig claimed in claim 1, it is characterized in that: the pH value of NaAc_HAc buffer solution is 4.5 ~ 6.
3. by wide spectrum immunostimulant preparation and preparation method for pig claimed in claim 1, it is characterized in that: the quality percent by volume of chitosan and NaAc_HAc buffer solution is 0.5 ~ 5%.
4. by wide spectrum immunostimulant preparation and preparation method for the pig described in claim 1 or 3, it is characterized in that: the molecular weight of the chitosan of selecting is at 200 ~ 600 kDa.
5. by wide spectrum immunostimulant preparation and preparation method for pig claimed in claim 1, it is characterized in that: the quality percent by volume of thymosin-interferon fusion protein and polyphosphoric acids sodium solution is 1 ~ 5%.
6. by wide spectrum immunostimulant preparation and preparation method for pig claimed in claim 1, it is characterized in that: the acetic acid-sodium acetate solution of chitosan-containing is 3:1 ~ 10:1 with the volume ratio of the polyphosphoric acids sodium solution containing thymosin-interferon fusion protein.
7. by wide spectrum immunostimulant preparation and preparation method for pig claimed in claim 1, it is characterized in that: the acetic acid-sodium acetate solution of chitosan-containing is 2 ~ 5 hours with the time of at room temperature reacting of the polyphosphoric acids sodium solution containing thymosin-interferon fusion protein.
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| CN201410062985.0A CN103877566A (en) | 2013-11-13 | 2014-02-25 | Swine broad-spectrum immune enhancement preparation and preparation method thereof |
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| CN201310561139.9A CN103599526A (en) | 2013-11-13 | 2013-11-13 | Pig broad spectrum immune enhancer and preparation method thereof |
| CN201310561139.9 | 2013-11-13 | ||
| CN201410062985.0A CN103877566A (en) | 2013-11-13 | 2014-02-25 | Swine broad-spectrum immune enhancement preparation and preparation method thereof |
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| CN201410062985.0A Pending CN103877566A (en) | 2013-11-13 | 2014-02-25 | Swine broad-spectrum immune enhancement preparation and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111281859A (en) * | 2019-12-23 | 2020-06-16 | 中国科学院海洋研究所 | Chitosan derivative nanoparticles with effect of enhancing antigen presenting cell to present antigen and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101347616A (en) * | 2007-05-18 | 2009-01-21 | 北京金迪克生物技术研究所 | Composing prescription of novel safe JY immunoadjuvant system and uses thereof |
| CN101361936A (en) * | 2008-09-18 | 2009-02-11 | 何进爱 | Traditional Chinese medicine combination of gynaecologic Shiyang washing lotion and preparation method thereof |
| CN101391103A (en) * | 2008-07-31 | 2009-03-25 | 张可 | Medicine composition for preventing and treating AIDS viral infection |
-
2013
- 2013-11-13 CN CN201310561139.9A patent/CN103599526A/en not_active Withdrawn
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- 2014-02-25 CN CN201410062985.0A patent/CN103877566A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101347616A (en) * | 2007-05-18 | 2009-01-21 | 北京金迪克生物技术研究所 | Composing prescription of novel safe JY immunoadjuvant system and uses thereof |
| CN101391103A (en) * | 2008-07-31 | 2009-03-25 | 张可 | Medicine composition for preventing and treating AIDS viral infection |
| CN101361936A (en) * | 2008-09-18 | 2009-02-11 | 何进爱 | Traditional Chinese medicine combination of gynaecologic Shiyang washing lotion and preparation method thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111281859A (en) * | 2019-12-23 | 2020-06-16 | 中国科学院海洋研究所 | Chitosan derivative nanoparticles with effect of enhancing antigen presenting cell to present antigen and preparation method thereof |
| CN111281859B (en) * | 2019-12-23 | 2022-03-22 | 中国科学院海洋研究所 | Chitosan derivative nanoparticles with effect of enhancing antigen presenting cell to present antigen and preparation method thereof |
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| CN103599526A (en) | 2014-02-26 |
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