CN107149674A - The purposes of IL-12 and flagellin in treatment chronic hepatitis B medicine is prepared - Google Patents
The purposes of IL-12 and flagellin in treatment chronic hepatitis B medicine is prepared Download PDFInfo
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- CN107149674A CN107149674A CN201610136820.2A CN201610136820A CN107149674A CN 107149674 A CN107149674 A CN 107149674A CN 201610136820 A CN201610136820 A CN 201610136820A CN 107149674 A CN107149674 A CN 107149674A
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- flagellin
- chronic hepatitis
- hbsag
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/208—IL-12
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/164—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
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- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
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Abstract
The invention discloses a kind of purposes of interleukin 12 and flagellin in treatment chronic hepatitis B medicine is prepared；It is the problem of HBsAg (+) (titre ＜ 1000IU/ml), HBeAg (), HBV DNA () chronic hepatitis B HBsAg are difficult to clean off to be intended to provide a kind of Testing index that solves, and new medicine is provided for chronic hepatitis B HBsAg；The purposes of technical points, interleukin 12 and flagellin in treatment chronic hepatitis B medicine is prepared, the Testing index of described chronic hepatitis B is：HBsAg+, titre ＜ 1000IU/ml, HBeAg, HBV DNA, the Testing index of described chronic hepatitis B is：HBsAg+, titre ＜ 1000IU/ml, HBeAg, HBV DNA.
Description
Technical field
It is IL-12 specifically the invention discloses a kind of IL-12 and the new application of flagellin
With purposes of the flagellin in treatment chronic hepatitis B medicine is prepared.
Background technology
There are 2,000,000,000 population infection hepatitis Bs in the whole world, wherein 2.4 hundred million be the viral surface antigen of chronic hepatitis B
(HBsAg) carrier.Chronic hepatitis B (CHB) some people is sluggish, but some people
Hepatic sclerosis, liver cancer can be caused.
In recent years, using interferon and nucleoside analogue treatment CHB, remarkable progress, Most patients are obtained
After treatment, the Cyklokapren of serum paddy third (ALT), HBVDNA, HBeAg can recover in peripheral blood
It is normal or switch to HBsAg still lasting masculins in feminine gender, but peripheral blood, it is extremely difficult to serological conversion and (is presented anti-
- HBs is positive).In these years, domestic and foreign scholars have proposed the concept that chronic hepatitis B feature is cured,
That is, chronic hepatitis B is treated in addition to requiring that ALT, HBVDNA, HBeAg are normal steadily in the long term,
Still require that peripheral blood HBsAg is eliminated, and it is positive by HBsAg hepatitis B surface antibody, i.e. serum occur
Be converted to feminine gender, and Anti-HBs antibody occur this is also terminal or target to treating chronic hepatitis B.Reach this mesh
Mark, is exactly that CHB features are cured.Chronic hepatitis B patient is due to the reduction of HBV specific immune functions
With CD8+T cell depletions, prevent its own from fully erased HBV, but it is produced to HBV antigens in vivo
The mainly Th1 type cells of raw response, therefore, Chronic Hepatitis B are improved using effective immunologic adjuvant
Cellular immune function, induces and maintains the cellullar immunologic response based on HBV specific CTLs, for
HBV thoroughly removing is particularly significant.
Flagellin (Piliated Pseudomonas aeruginosa, PPA) can adjust human body fluid it is immune and
The activity of the non-equilibrium state of cellular immunity, increase macrophage and NK cells, maintains the quantity of T cell
With ratio.Flagellin contains I, II, IV type pili and flagellin, the elder generation that flagellin starts simultaneously
Its innate immune response is mediated by TLR5, is produced various biologicallies, is congenital innate immune response
With the key protein in day after tomorrow Acquired immune response.TLR5 has been demonstrate,proved as one of member of TLR families
It is bright main in lungs and liver expression.
Interleukin 12 (IL-12), as the core cell factor in immunological network, is that connection is congenital intrinsic
The immune bridge with the acquired immunity day after tomorrow, can significantly induce Th1 type cellular immunities, and enhancing CD8+T is thin
The cytotoxicity of born of the same parents, and suppress Th2 type cellular immunities.These characteristics make it suppress the mistake of hbv replication
Played a significant role in journey.IL-12, which has, promotes T cell activation, propagation, the function of cracking effect, has
Hoping turns into effective immunopotentiator, to aid in the immune response of CD8+T cells.
In present treatment of chronic, HBsAg (+) (titre ＜ 1000IU/ml), HBeAg (-),
HBV-DNA (-) is difficult to clean off surface antigen.
The content of the invention
In view of the above-mentioned problems, it is HBsAg (+) (drops to solve Testing index it is an object of the invention to provide one kind
Degree ＜ 1000IU/ml), HBeAg (-), HBV-DNA (-) chronic hepatitis B HBsAg be difficult to
The problem of removing, is used as the use prepared in treatment chronic hepatitis B medicine there is provided IL-12 and flagellin
On the way.
In order to solve the above technical problems, the technical scheme that the present invention is provided is such：
The purposes of a kind of IL-12 and flagellin in treatment chronic hepatitis B medicine is prepared.
Further, above-mentioned IL-12 and flagellin are in treatment chronic hepatitis B medicine is prepared
Purposes, the Testing index of described chronic hepatitis B is：HBsAg+, titre ＜ 1000IU/ml, HBeAg-,
BV-DNA-。
Further, above-mentioned IL-12 and flagellin are in treatment chronic hepatitis B medicine is prepared
Purposes, described flagellin dosage contains bacterium 1.0 × 10 for every dose9-3.0×109；Described interleukins
- 12 dosage are 5-10 μ g/ agent.
Compared with existing, the present invention is provided color technical scheme and joined using interleukin 12 with flagellin preparation
Conjunction is used as treatment HBsAg (+) (titre ＜ 1000IU/ml), HBeAg (-), HBV-DNA
The medicine of the chronic hepatitis B of (-)；Wherein, flagellin joint rhIL-12 can significantly improve Healthy People
The level of IFN-γ is produced with chronic hepatitis B patient PBMCs, strengthens its cellullar immunologic response, is promoted
The removing of chronic hepatitis B surface antigen.
Brief description of the drawings
Fig. 1 is the level that various concentrations flagellin joint rhIL-12 induces IFN-γ；
Fig. 2 is the level that flagellin joint various concentrations rhIL-12 induces IFN-γ；
Fig. 3 flagellins combine IFN-γ in rhIL-12 inducing chronics hepatitis B patient and Healthy People PBMCs
Level；
Fig. 4 flagellins combine effects of the rmIL-12 to hepatitis B hydrodynamic model mice serum HBsAg.
Embodiment
With reference to embodiment, the claim to the present invention is described in further detail, but not
Any limitation of the invention is constituted, any limited number of time done in the claims in the present invention protection domain
Modification, still within the claims of the present invention.
Embodiment 1
The interleukin 12 that the present invention is provided is used for HBsAg (+) (titre ＜ as a kind of medicine
1000IU/ml), HBeAg (-), the treatment of HBV-DNA (-) chronic hepatitis B, when using and whip
Hairless protein agents are used, and flagellin joint rhIL-12 can significantly improve Healthy People and chronic type b liver
Scorching patient PBMCs produces the level of IFN-γ, strengthens its cellullar immunologic response, promotes chronic hepatitis B
The removing of surface antigen.
Specific experiment example is as follows：
The selection of flagellin dose,optimum：Flagellin group (0.0075~14ng/ml), rhIL-12
(1ng/ml), various concentrations flagellin+rhIL-12, negative control group (cell culture fluid), the positive are right
According to group (the anti-human CD3 of 0.2ug/ml), respectively with PMNC (PBMCs), cell concentration
For 2 × 106/ ml, 37 DEG C, 5%CO2It is incubated IFN-γ in 72h, ELISA method detection cells and supernatant
Content.
As a result：The generation of IFN-γ in flagellin joint rhIL-12 induction Healthy Peoples PBMCs, certain
In the range of increase with the rise of flagellin concentration, optimal dilution scope be 1.75-14ng/ml, see Fig. 1.
Independent flagellin group can not induce the generation of IFN-γ substantially, similar to negative control group；It is used alone
RhIL-12 (1ng/ml) can also produce certain stimulation.
The selection of rhIL-12 dose,optimums：RhIL-12 (concentration is respectively 0.1,0.5,1.0 and 2.0ng/ml),
Flagellin (3.5ng/ml), flagellin+various concentrations rhIL-12 groups, negative control group (cell culture
Liquid), positive controls (the anti-human CD3 of 0.2ug/ml), respectively with PMNC (PBMCs),
Cell concentration is 2 × 106/ ml, 37 DEG C, 5%CO2It is incubated in 72h, ELISA method detection cells and supernatant
The content of IFN-γ.
As a result：The selection of rhIL-12 dose,optimums：Flagellin joint rhIL-12 induction Healthy Peoples PBMCs
The generation of middle IFN-γ, increases, optimum concentration scope is with the rise of rhIL-12 concentration within the specific limits
0.1-1.0ng/ml；RhIL-12, which is used alone, can also produce stimulation.See Fig. 2.
Flagellin combines the generation of IFN-γ in rhIL-12 inducing chronics hepatitis B patient and Healthy People PBMCs：
RhIL-12 (concentration is respectively 0.1,0.5,1.0 and 2.0ng/ml), flagellin (3.5ng/ml), flagellum
Albumen+various concentrations rhIL-12 groups, negative control group (cell culture fluid), positive controls (0.2ug/ml
Anti-human CD3), respectively with PMNC (PBMCs), cell concentration is 2 × 106/ ml, 37 DEG C,
5%CO2It is incubated the content of IFN-γ in 72h, ELISA method detection cells and supernatant.
As a result：Flagellin combines IFN-γ in rhIL-12 inducing chronics hepatitis B patient and Healthy People PBMCs
Generation：Flagellin group only induces the low-level IFN-γ of generation, and the induction of rhIL-12 groups produces IFN-γ
Level is significantly improved compared with negative control group；And the level that flagellin+rhIL-12 group inductions produce IFN-γ shows
Write and be higher than flagellin and rhIL-12 groups, difference is statistically significant.See Fig. 3.
Flagellin combines influences of the rmIL-12 to hepatitis B hydrodynamic model mice serum HBsAg titres:32
Hepatitis B hydrodynamic model mouse is randomized into physiological saline group, rmIL-12 (100ng/ is only), flagellin
(μ g/ are only) and flagellin+rmIL-12 groups, every group 8.RmIL-12 weekly administrations twice, for the first time
It is administered once again after administration 72h, successive administration three weeks；Flagellin weekly administration once, successive administration three
Week.Injection site is that mouse nape part is subcutaneous.Respectively at blood sampling is surveyed in serum weekly before administration, after administration
HBsAg titre, continues to observe three weeks, the titre of HBsAg in serum is surveyed in blood sampling weekly after drug withdrawal.
As a result：Flagellin combines effects of the rmIL-12 to hepatitis B hydrodynamic model mice serum HBsAg：
Flagellin group model mice serum HBsAg declines with being compared before administration in low-level；RmIL-12 group moulds
Type mice serum HBsAg is remarkably decreased with being compared before administration, and HBsAg starts knock-on after drug withdrawal；And flagellum
Albumen+rmIL-12 group model mice serum HBsAg before administration with being compared less than flagellin and rmIL-12
Group, difference is statistically significant, and HBsAg continuous decreases after drug withdrawal, has no knock-on.See Fig. 4.
As a result show：HBsAg specificity P peptides joint IL-12, which has, significantly promotes HBsAg serological conversions to act on.
The present invention provides a kind of interleukin 12 joint flagellin treatment HBsAg (+) (titre ＜
1000IU/ml), HBeAg (-), HBV-DNA (-) chronic hepatitis B, it can effectively be opened
Move the immune CD8+T cells exhausted with the acquired immunity day after tomorrow, recovery of congenital inherency, the single core of peripheral blood
Cell (PBMCs) produces the level of IFN γ, removes HBsAg.
Above-described is only presently preferred embodiments of the present invention, all institutes in the range of the spirit and principles in the present invention
Any modifications, equivalent substitutions and improvements made etc., should be included in the scope of the protection.
Claims (3)
1. a kind of purposes of IL-12 and flagellin in treatment chronic hepatitis B medicine is prepared.
2. IL-12 according to claim 1 and flagellin are preparing treatment chronic hepatitis B
Purposes in medicine, it is characterised in that the Testing index of described chronic hepatitis B is：HBsAg+, drop
Spend ＜ 1000IU/ml, HBeAg-, HBV-DNA-.
3. IL-12 according to claim 1 and flagellin are preparing treatment chronic hepatitis B
Purposes in medicine, it is characterised in that the dosage of described flagellin is every dose and contains bacterium 1.0 × 109-3.0
×109；The dosage of described interleukin 12 is 5-10 μ g/ agent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610136820.2A CN107149674A (en) | 2016-03-10 | 2016-03-10 | The purposes of IL-12 and flagellin in treatment chronic hepatitis B medicine is prepared |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610136820.2A CN107149674A (en) | 2016-03-10 | 2016-03-10 | The purposes of IL-12 and flagellin in treatment chronic hepatitis B medicine is prepared |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107149674A true CN107149674A (en) | 2017-09-12 |
Family
ID=59791990
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610136820.2A Pending CN107149674A (en) | 2016-03-10 | 2016-03-10 | The purposes of IL-12 and flagellin in treatment chronic hepatitis B medicine is prepared |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107149674A (en) |
-
2016
- 2016-03-10 CN CN201610136820.2A patent/CN107149674A/en active Pending
Non-Patent Citations (4)
| Title |
|---|
| RISINI D. WEERATNA: "TLR agonists as vaccine adjuvants_ comparison of CpG ODN and Resiquimod (R-848)", 《VACCINE》 * |
| 傅泳航: "绿脓杆菌及其鞭毛蛋白联合rhIL-12体外对慢性乙型肝炎患者细胞免疫功能的影响", 《中国生物制品学杂志》 * |
| 张辉: "嵌合鞭毛蛋白fliC esat佐剂效应的研究", 《细胞与分子免疫学杂志》 * |
| 李琴: "鞭毛蛋白与IL-12协同诱导人NK细胞产生IFN-γ机制的探讨", 《中国免疫学杂质》 * |
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Application publication date: 20170912 |