Pharmaceutical composition and uses thereof

A composition and drug technology, applied in drug combinations, antitumor drugs, pharmaceutical formulations, etc., can solve problems such as unsatisfactory, rare, and negative effects, and achieve the prevention of tumor occurrence, less toxic and side effects, and a wide antigen spectrum. Effect

Inactive Publication Date: 2014-06-18
FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although there are many related studies on tumor vaccines at home and abroad, so far few breakthroughs have been made. The main problems are: the technology of introducing DNA into specific cells for expression is not yet mature, and the use of exogenous The safety of DNA has not yet been resolved; due to the high heterogeneity of tumor cells, multiple tumor cells belonging to the same type of tumor can express different antigens, and T cells activated by one tumor antigen can only kill a part of tumor cells , tumor cells that do not express this antigen cannot be killed; although tumor cell vaccines can contain almost all tumor antigens, current studies have shown that their role in activating T cells is limited, and the effect of using them as a vaccine is not ideal
[0009] However, in recent years, some related experiments have shown that some tumor-derived exosomes can inhibit or even destroy immune cells that play a role in tumors, such as down-regulating the expression of some NK receptors, affecting the activation of some innate immune cells in tumor immunity , and some can significantly inhibit IL-2 to inhibit the proliferation of human lymphocytes, thus playing some negative roles in tumor immunotherapy
These tumor-derived exosomes may be the key factor for tumor tissue to escape the body's immune system clearance, which brings many difficulties and challenges to tumor immunotherapy

Method used

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  • Pharmaceutical composition and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Cell Culture

[0039] Experimental materials: H22 cell line, 10% fetal bovine serum, penicillin and streptomycin mixture (Beijing Tigemei Company).

[0040] Experimental method: culture H22 cell line in the culture medium containing 10% fetal bovine serum, penicillin 100IU / mL, streptomycin 100×μg / mL, at 37°C and 5% CO 2 Culture in the incubator, when the cells grow to the logarithmic phase, press 3×10 6 / 100ml inoculation.

Embodiment 2

[0041] Embodiment 2 drug treatment

[0042] Experimental materials: procaine, MS-275 (both purchased from sigma company).

[0043] Experimental grouping: the volume of cell culture medium is strictly consistent in each group, blank control group, exosomes control group (no drug addition group), exosomes experimental group 1 (adding procaine), exosomes experimental group 2 (adding drug MS-275) , exosomes experimental group 3 (combined administration of procaine and MS-275).

[0044] Experimental method: in the control group, no drug was added after inoculation, and 150ml of the supernatant was collected as a control after 24 hours; -6 mol / L of procaine, respectively collected 150ml of culture supernatant 24h after dosing; dosing group 2, 24h after inoculation with a concentration of 1 × 10 -6 mol / L MS-275 treatment, respectively collected 150ml of culture supernatant 24h after dosing; dosing group 3, 24h after inoculation with 1 × 10 -6 mol / L procaine and 1×10 -6 mol / L MS-2...

Embodiment 3

[0045] The separation and purification of embodiment 3 exosomes

[0046] Experimental equipment: HMAC-CP70G low temperature ultra-high speed centrifuge, 100KU MW CO Millipore Amicon high recovery rate high flow rate tangential flow ultrafiltration centrifuge tube (Millipore Company).

[0047] Experimental method: Centrifuge the collected cell culture supernatants of the experimental group and the control group at 300g for 10min to remove the cells, and take the supernatant; centrifuge at 1500g for 30min to remove the cell debris, collect the supernatant, and concentrate it through a 100kUMWCO Centriplus centrifugal ultrafiltration tube Ultrafiltration, centrifugation at 1500g for 30min to obtain 6ml of concentrated solution, transfer the separated and purified concentrated solution to a 1.5ml centrifuge tube, and ultracentrifuge at 100kg for 60min at 4°C with a horizontal rotation angle to obtain exosomes.

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Abstract

The invention discloses a pharmaceutical composition and uses thereof. The pharmaceutical composition comprises a DNA methyltransferase inhibitor and a histone deacetylase inhibitor. The invention also discloses new uses of the pharmaceutical composition. The pharmaceutical composition is used to treat tumor cells together to obtain an exosomes tumor vaccine. The invention also discloses a method for preparing a tumor vaccine by using the pharmaceutical composition. The method comprises the following steps of using procaine and MS-275 to treat the tumor cells, and separating and purifying exosomes secreted by the tumor cells. The pharmaceutical composition disclosed by the invention improves the therapeutic effect of the exosomes tumor vaccine, and has an important clinical application value.

Description

technical field [0001] The present invention relates to a pharmaceutical composition and its application, in particular to a pharmaceutical composition and its new application in the preparation of tumor vaccines, more specifically to a pharmaceutical composition comprising procaine and MS-275 and its application Use in the preparation of exosomes tumor vaccines. Background technique [0002] Procaine, chemical name 2-(diethylamino)ethyl 4-aminobenzoate, is a common local anesthetic that has been used clinically for nearly a hundred years. The drug can stabilize the cell membrane and reduce its The permeability of ions prevents sodium and potassium ions from entering and leaving the cell membrane to generate depolarization and action potential when the nerve impulse arrives, thereby producing local anesthesia. In addition, procaine is also widely used in the treatment of many diseases in various clinical departments, such as the clinical treatment of pertussis in pediatrics...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/06A61K38/55A61K39/00A61K39/39A61P35/00A61K31/245
Inventor 肖文华董伟伟
Owner FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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