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A combined medicine capable of improving the therapeutic effect of polypeptide vaccine on HPV infected tumor and application thereof

A peptide vaccine and tumor treatment technology, applied in the field of biomedicine, can solve the problem of not improving the survival time of mice, and achieve the effect of extensive immunogenicity and easy production

Active Publication Date: 2019-01-15
ZHONG AO BIOMEDICAL TECH (GUANGDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] However, neither the F1 and F3 polypeptide compositions disclosed in Chinese invention patent 20161122304.8 nor the therapeutic vaccine immunization disclosed in Chinese invention patent 201810560842.0 can improve the survival time of tumor-bearing mice alone

Method used

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  • A combined medicine capable of improving the therapeutic effect of polypeptide vaccine on HPV infected tumor and application thereof
  • A combined medicine capable of improving the therapeutic effect of polypeptide vaccine on HPV infected tumor and application thereof
  • A combined medicine capable of improving the therapeutic effect of polypeptide vaccine on HPV infected tumor and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0052] The preparation method of F1, F3 polypeptide cream of the present invention is as follows: Poloxamer 407 (molecular weight 12600, pH 6-7.4, batch number WPAK592B, BASF, Germany), poloxamer 188 (molecular weight 8400, pH 6-7, batch number WPAK539B, BASF, Germany) ;

[0053] Blank gel preparation: Add 46 grams of poloxamer 407 and 10 grams of poloxamer 188 into 200ml of distilled water, place at 4 degrees Celsius until poloxamer 407 and 188 are completely dissolved; stir well until the gel is formed. Add 10mg of F1 or F3 to 10ml blank gel. After complete dissolution, filter with a 0.22um filter.

[0054] The four polypeptides in the polypeptide composition of the present invention (hereinafter referred to as EX) are artificially synthesized by a biotechnology company using the Fmoc-Glu solid-phase method, and the crude peptides are purified by high performance liquid chromatography with a purity of >95% and freeze-dried at -20 ℃ and save for future use.

Embodiment 1

[0055] Example 1: Polypeptide composition combined with different Toll-like receptor ligands, or combined with aIL10R immunization

[0056] Experimental procedure

[0057] C57BL / 6 mice were randomly divided into 4 groups, group 1 was EX+MPLA; group 2 was EX+MPLA+Rat IgG; group 3 was EX+MPLA+aIL10R; group 4 was PBS control group.

[0058] The four groups of mice were subcutaneously immunized on the left abdomen, once a week, and immunized twice in total. The mice were sacrificed on the 6th day after the last immunization, the spleens of the mice were removed, and the antigen-specific CD8+ T cell response induced by the vaccine was detected by ELISPOT method.

[0059] the result shows:

[0060] 1. The 4 E7 polypeptide / MPLA / anti-interleukin 10 receptor antibody vaccine containing the full length of HPVE7 induces more antigen-specific CD8+ T cells than the vaccine without the interleukin 10 receptor antibody vaccine. See results figure 1 As shown in A.

[0061] 2. C57BL / 6 mic...

Embodiment 2

[0063] Example 2 F1 and F3 compositions can increase the effect of polypeptide vaccines with specific immune function in inhibiting the growth of TC-1 tumors

[0064] Experimental procedure

[0065] 1. Cultured in 100 microliters of 5X10 in RPMI medium containing 10% calf serum 3 TC-1 cells were added to a 96-well cell culture plate, and 5 micrograms per milliliter of F1 or / and F3 and control polypeptide P3 were added, and heated at 37°C with 5% CO 2 After culturing overnight, the supernatant was taken, and MCP-1 was detected by ELISA. F1 and F3 compositions can stimulate TC-1 cells to secrete more MCP1, the results can be found in figure 2 .

[0066] 2. Inoculate TC-1 tumor subcutaneously into C57BL / 6 mice or nude mice. After the tumor can be touched, inject F1 and F3 compositions into the tumor 7 times, 30 μg / time, Imiquimod 50 μg / time, and kill the mice two days later. Rats were isolated and weighed. Compositions F1 and F3 can inhibit the growth of TC-1 tumor in mice....

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Abstract

The invention discloses a combined medicine for improving the therapeutic effect of a polypeptide vaccine on benign and malignant tumors caused by human papillomavirus (HPV) infection and applicationthereof. The combined medicine comprises at least (i) a polypeptide vaccine having a specific immune function and (ii) a polypeptide composition comprising F1, F3. The F1, F3 polypeptide composition can stimulate tumor cell to secrete inflammatory factor MCP-1, attracting inflammatory cells such as T cells and NK cells into tumor tissues. The combined medicine can inhibit TC-1 tumor cell growth inmice. The F1, F3 polypeptide composition can improve that efficiency of therapeutic vaccines in the present application and can prolong the survival time of tumor-bearing mice immunize with therapeutic vaccines. The combination of the polypeptide vaccine and the F1, F3 polypeptide composition according to the effective dosage ratio is more effective than the combination of the polypeptide vaccineand the F1, F3 polypeptide composition alone, and has synergistic effect.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a combined drug capable of improving the therapeutic effect of a polypeptide vaccine on tumors related to HPV infection and its application. Background technique [0002] Chronic human papillomavirus (human papillomavirus, HPV) infection can cause a variety of diseases. Cancers of the cervix, vulva, penis, anus, and head and neck cancers, including oral cancer, can all be caused by infection with HPV. Cervical cancer is one of the common malignant tumors, and its incidence rate is the second among female tumors. Its occurrence is closely related to the persistent infection of HPV, especially HPV types 16 and 18. Although vaccines for the prevention of HPV infection have been used since 2006, such vaccines are only suitable for healthy individuals who have never been infected, and are ineffective against current HPV infection and the diseases it causes. At presen...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61K39/12A61K39/39A61K39/395A61P31/20A61P35/00
CPCA61K38/1703A61K39/12A61K39/39A61K39/3955A61P31/20A61P35/00A61K2039/54A61K2039/55511A61K2039/55516A61K2039/55522A61K2039/55561A61K2039/55572A61K2039/55544A61K2039/585C12N2710/20034A61K2300/00
Inventor 刘晓松王天放倪国颖
Owner ZHONG AO BIOMEDICAL TECH (GUANGDONG) CO LTD
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