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Pharmaceutical composition and application thereof

A composition and drug technology, applied in the field of chickenpox or post-herpetic neuralgia, prevention of herpes zoster, can solve problems such as huge differences in effects, and achieve the effects of excellent effects, superior immune stimulation, and high antibody levels

Pending Publication Date: 2021-06-18
JIANGSU THERAVAC BIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the structural diversity of CpG oligodeoxynucleotides, the effects of CpG adjuvants of different sequences vary greatly

Method used

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  • Pharmaceutical composition and application thereof
  • Pharmaceutical composition and application thereof
  • Pharmaceutical composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Preparation method of recombinant herpes zoster vaccine composition

[0056] 1.1 Preparation of herpes gE protein: the amino acid sequence is shown in SEQ ID NO:1.

[0057] References Thomsson E, Persson L et al. report in "Journal of Virological Methods" 2011, volume 175, phase 1, pages 53-59, the specific steps are as follows:

[0058] Optimize the nucleic acid sequence according to the target protein sequence so that its codons conform to the mammalian expression system, and synthesize the target gene; connect the synthesized target gene to the pcDNA3.1(+) plasmid by enzyme digestion and ligation, and transform Top 10 competent, pick positive single clones, and perform sequencing verification on positive single clones; a large number of single clones are amplified, and a large number of plasmids that meet cell transfection are extracted with an endotoxin-free plasmid extraction kit; Transfection method: Transfect CHO suspension cells with the plasmid. Wh...

Embodiment 2

[0064] Example 2 Screening experiment of CPG oligodeoxynucleotides

[0065] 2.1 Experimental animals:

[0066] C57BL / 6(N) mice, 42 females, 5 weeks old, were purchased from Shanghai Slack Experimental Animal Co., Ltd.

[0067] 2.2 Reagent materials:

[0068] The herpes gE stock solution obtained in Example 1 was diluted to 50 μg / mL with PBS solution (purchased from Hyclone), and the CpG obtained in Example 1 was diluted to 100 μg / mL with PBS solution.

[0069] 2.3 Experimental grouping:

[0070] See Table 2, the injection volume of each mouse was 100 μL / mouse, and the control group was injected with PBS solution 100 μL / mouse.

[0071] Table 2 Grouping of experimental animals and the amount injected in each group

[0072]

[0073] 2.4 Animal immunity:

[0074] All groups were injected intramuscularly every 2 weeks, and the inoculation site was the right rear thigh, and the mice were administered twice in a row, that is, at week 0 and week 2, respectively, and all mice...

Embodiment 3

[0084] Example 3 Effects of different doses of immunostimulatory compositions on the efficacy of recombinant herpes zoster vaccine compositions

[0085] 3.1 Experimental animals and model establishment:

[0086] C57BL / 6(N) mice, 72 female, 5 weeks old, were purchased from Shanghai Slack Experimental Animal Co., Ltd.

[0087] 3.2 Reagent materials:

[0088] 1) Herpes gE protein, CpG T1, and CpG 7909 were all obtained from Example 1;

[0089] 2) QS-21 (CAS.NO.A010-023, purchased from BRENNTAG);

[0090] 3) Use PBS solution (purchased from Hyclone) to dilute herpes gE stock solution to 50 μg / mL, use PBS solution to dilute QS-21 to 5 μg / mL, 50 μg / mL, and 100 μg / mL, and use PBS solution to dissolve CpG T1 And dilute to 50μg / mL, 100μg / mL, 2mg / mL respectively, use PBS solution to dissolve CPG7909 and dilute to 100μg / mL for the next step.

[0091] 3.3 Experimental grouping:

[0092] See Table 4, the volume of each injection is 100 μL / monkey. The control group was injected with...

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Abstract

The present invention provides a pharmaceutical composition. The pharmaceutical composition comprises i) a herpes gE protein, an active fragment of the protein, a variant of the protein, or a mixture of at least two of the herpes gE protein, the active fragment of the protein and the variant of the protein; Ii) an immunostimulatory composition, wherein the immunostimulatory composition comprises a saponin and a CpG oligodeoxynucleotide, or the immunostimulatory composition comprises an adjuvant comprising the saponin, and the CpG oligodeoxynucleotide. The invention provides application of the pharmaceutical composition in preparation of a medicine for preventing and / or treating chicken pox-herpes zoster virus infection and / or chicken pox-herpes zoster virus mediated diseases. The pharmaceutical composition provided by the invention achieves an unexpected technical effect and can mediate stronger immune response.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals. Specifically, the present invention relates to a pharmaceutical composition comprising herpes gE protein and an immunostimulatory composition. Wherein, the immunostimulatory composition comprises saponin and CpG oligodeoxynucleotide, or the immunostimulatory composition is composed of an adjuvant comprising saponin and CpG oligodeoxynucleotide, wherein the CpG oligodeoxynucleotide The nucleotide sequence has two or more copies of the 5'-TTCGTT-3' motif or the 5'-TCGTCGTCG-3' motif. The present invention also relates to the use of the pharmaceutical composition for preventing herpes zoster, chickenpox or postherpetic neuralgia. Background technique [0002] Herpes zoster is a viral skin disease caused by varicella-zoster virus (VZV). During primary infection, VZV enters the local lymph nodes through the respiratory mucosal epithelium and replicates. The virus-infected lymphocytes then enter th...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K39/00A61P31/00A61P33/00A61P35/00A61P37/04
CPCA61K39/39A61K39/00A61P31/00A61P33/00A61P35/00A61P37/04A61K2039/55561A61K2039/55577Y02A50/30A61P31/20A61K39/12A61K2039/572A61K2039/575A61K2039/70C12N2710/16734A61P31/12A61P31/04A61K39/245A61K39/29A61P31/22A61K31/711C07K16/2803A61K39/292
Inventor 葛君李建强孙娇娇任苏林周童王世伟王晓东黄精俸陈悦
Owner JIANGSU THERAVAC BIO PHARMA
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