Pharmaceutical composition containing rhIL-12 for treating inactive hepatitis B

A composition and active technology, applied in the field of biological preparations, can solve the problems of unable to form immune memory, unable to clear HBV well, unable to completely clear, etc., to improve acquired immune function, improve cellular immunity and humoral immune function , the effect of strong cellular immune enhancement

Active Publication Date: 2016-12-21
GUANGDONG COOWAY BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of acquired immunity, due to the persistent presence of HBV, the programmed death-1 receptor (PD1) and its ligand (PDL-1), which negatively regulate the immune response, are highly expressed in CD4+ and CD8+ T cells , causing T cells to transform into exhausted T cells (T cellexhausted), that is, non-functional T cells, so that they cannot clear HBV well, let alone form immune memory, so that they cannot monitor the production of HBsAg, resulting in the persistence of HBsAg and the persistent presence of HBsAg in serum. positive
Therefore, under current treatment methods, HBsAg persists and cannot be completely cleared

Method used

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  • Pharmaceutical composition containing rhIL-12 for treating inactive hepatitis B
  • Pharmaceutical composition containing rhIL-12 for treating inactive hepatitis B
  • Pharmaceutical composition containing rhIL-12 for treating inactive hepatitis B

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 The effect of rhIL-12 on the induction of IFN-γ by PBMCs of healthy people and CHB patients

[0021] 1. Experimental data: rhIL-12 in this example was purchased from Guangzhou Kaitai Biotechnology Co., Ltd.; human IFN-γELISA kit was purchased from BD / Pharmingen (Becton Dickinson / Phar Mingen USA); RPMI-1640 culture medium , glutamine and Hanks solution were purchased from GIBCO (USA); dextran Ficoll-Hypaque was purchased from Shanghai Huajing Biotechnology Company; fetal bovine serum was purchased from GIBCO.

[0022] 20 cases of healthy people and 62 cases of CHB patients were screened (including 21 cases in the immune tolerance phase, 22 cases in the immune clearance phase, and 19 cases in the inactive HBsAg carrier phase). Among them, the selection criteria for healthy people: HBsAg negative, HBsAb positive. All hepatitis B patients were not combined with hepatitis C infection, and all subjects were aged 15-55, regardless of gender.

[0023] 2. Test method...

Embodiment 2

[0043] Example 2 Effects of combined application of rhIL-12 and specific antigens on the production of IFN-γ in BALB / c mice

[0044] 1. Experimental data: rhIL-12 in this example was purchased from Guangzhou Kaitai Biotechnology Co., Ltd.; RPMI1640 basal culture medium and FBS were purchased from GIBCO (USA); Ficoll lymphocyte separation medium was purchased from Shanghai Huajing Biotechnology company.

[0045] 40 clean-grade BALB / c mice, half male and half female, weighing 20 g, were purchased from the Experimental Animal Center of Southern Medical University.

[0046] 2. Test method:

[0047] 40 mice were randomly divided into normal saline group (normal saline 150 μl / mouse), rHBsAg group (10 μg / mouse), Mycobacterium vaccae group (10 μg / mouse), rhIL-12 group (0.3 μg / mouse), rHBsAg (10 μg / monkey)+Mycobacterium vaccae (10 μg / bird)+rhIL-12 (0.3 μg / bird) group, a total of five groups, 8 rats in each group, half male and half male. Dosing twice, and dosing again after 72 hours...

Embodiment 3

[0055] Example 3 The combined application of rhIL-12 and specific antigen on the production of IFN-γ by PBMCs of healthy people and CHB patients

[0056] 1. Experimental data: rhIL-12 in this example was purchased from Guangzhou Kaitai Biotechnology Co., Ltd.; rHBsAg was purchased from Shenzhen Kangtai Biotechnology Co., Ltd.; Mycobacterium vaccae for injection was purchased from Anhui Zhifeilong Kema Biopharmaceutical Co., Ltd. Ltd.; human IFN-γELISA kits were purchased from BD Company.

[0057] Screening 15 cases of chronic hepatitis B patients, aged 17-49 years, with an average of 31.17 years old. Acute and chronic diseases caused by hepatitis A virus (HAV), hepatitis C virus (HCV), hepatitis D virus (HDV), hepatitis E virus (HEV) infection and other causes (drugs, alcohol, poisoning) were excluded for all patients Liver damage. Another 9 healthy volunteers were selected as controls, all of whom had been vaccinated with recombinant HBsAg vaccine, except for HBsAb positive...

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Abstract

The invention relates to the field of biological preparations, in particular to a pharmaceutical composition containing rhIL-12 for treating inactive hepatitis B. The pharmaceutical composition, in terms of each milliliter, consists of 10-20 [mu]g of recombinant human interleukin-12, 10-60 [mu]g of a recombinant HBsAg protein vaccine, 5-20 [mu]g of a cow mycobacteria vaccine and the balance of a water solution. The pharmaceutical composition is capable of activating a T achroacyte subgroup, promoting CD3+CD4+ and CD3+CD8+ cells to secrete IFN-[gamma] and inhibiting PD-1 expression of the CD3+CD4+ and CD3+CD8+ cells; in addition, the pharmaceutical composition can also activate mononuclear cells and dendritic cells, promote CD14+ cells and CD1a+ cells to express Toll-like receptors and enhance specific immunity and non-specific immunity functions, so as to break through an immune tolerance state and to better cure HBV infection; and in addition, the pharmaceutical composition can promote HBsAg seronegative conversion.

Description

technical field [0001] The invention relates to the field of biological preparations, in particular to a pharmaceutical composition containing rhIL-12 for treating inactive hepatitis B. Background technique [0002] Hepatitis B (hepatitis B) is caused by hepatitis B virus (HBV) infection. It is a common disease in my country and has certain infectivity. Among them, hepatitis B patients and HBV carriers are the main sources of infection for this disease. The clinical manifestations are fatigue, fear of eating, nausea, abdominal distension, pain in the liver area and other symptoms. In severe cases, it can develop into liver cirrhosis and liver cancer, which seriously affects the health of our people. [0003] Currently, hepatitis B treatment mainly includes antiviral, immune regulation, antifibrosis and symptomatic treatment. Drugs such as nucleoside analogues, interferon-α (IFN-α) and long-acting interferon (PEG IFN-α) are often used alone or in combination. This treatment ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/29A61K38/20A61K39/39A61K9/08A61K9/19A61P31/20
CPCA61K9/0019A61K9/08A61K9/19A61K38/208A61K39/12A61K39/39A61K2039/55594C12N2730/10134A61K2300/00
Inventor 陈松彬倪彦艳李小翠
Owner GUANGDONG COOWAY BIOTECH CO LTD
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