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Medicine as well as preparation method and application thereof

A drug and reaction technology, applied in drug combinations, pharmaceutical formulations, anti-tumor drugs, etc., can solve the problem that anti-tumor DNA or peptide vaccines cannot truly enter the clinic, low antigen capture and uptake ability, and unsatisfactory anti-tumor vaccine immune effects, etc. question

Active Publication Date: 2017-08-18
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Studies have shown that the main reason for the unsatisfactory immune effect of anti-tumor vaccines in vivo is that plasmid DNA or polypeptides are degraded in large quantities before entering cells, and immature dendritic cells (Dendritic cells, DC) of tumor patients The uptake ability of antigen capture is low, which is one of the main reasons why many existing anti-tumor DNA or peptide vaccines cannot really enter the clinic

Method used

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  • Medicine as well as preparation method and application thereof
  • Medicine as well as preparation method and application thereof
  • Medicine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] The preparation of embodiment 1 carrier

[0070] Taking polyurethane compound as an example, its molecular structure is:

[0071]

[0072] The specific method of polymer synthesis is: (1) Weigh 1.2mmol of 1,6-hexanediol diacrylate, 0.9mmol of N,N-dibutylpropylenediamine and 0.1mmol of NH2-mPEG2000, and dissolve them all Disperse in 2ml of dimethyl sulfoxide (DMSO), pass through N after completely dissolving 2 15 minutes, then heated to 50°C and reacted in the dark for 7 days; (2) Take out the reaction product after 7 days, use a dialysis bag with a cut-off of 3500 to dialyze in water for 6-9 hours, and collect the product in the dialysis bag; (3) Freeze-dried, the product was collected after freeze-drying for subsequent experiments.

[0073] The NMR structure of the carrier is as figure 2 As shown, the position of the dotted box indicates a double bond, and it can be seen that the vector was successfully constructed.

Embodiment 2

[0074] The preparation of embodiment 2 medicines

[0075] Dissolve the reaction product in Example 1 in DMSO, add excess autophagy-inducing peptide Beclin1, and tumor antigen peptide OVA, and react in the dark at 37°C for 3 days; (2) Take out the reaction product after 3 days, and use a cut-off of The dialysis bag of 3500 was dialyzed in water for 6-9 hours, and the product in the dialysis bag was collected; (3) freeze-dried, and the product was collected after freeze-drying for subsequent experiments. The product is a molecule with autophagy induction function and tumor antigen loading.

[0076] The obtained modified polypeptide molecule, that is, the chemical structure of the drug molecule 3 is as follows:

[0077]

[0078] Its schematic diagram is as follows figure 1 As shown, the NMR spectrum is as image 3 As shown, it can be seen from the figure that the double bond at both ends of the polymer in the original carrier molecule disappears (the dotted box indicates th...

Embodiment 3

[0081] Drugs described in Example 3 improve the induction of autophagy

[0082] First, use the DC2.4 cell line as the object to study the effect of molecularly induced autophagy. Divide DC2.4 cells into 5×10 4 Inoculate confocal dishes at 37 °C and 5% CO 2 Incubate for 12 hours under the conditions; use lipo3000 to transfect the cells, transfect the GFP-LC3 plasmid, and after 6 hours of transfection, replace the medium containing the transfection reagent with complete medium, and culture for another 12-16 hours;

[0083] Add autophagy inducers, inhibitors and the corresponding molecule 3 or control molecule 2 to the transfected cells respectively. After incubation for 12 hours, wash the cells with PBS and observe the GFP fluorescence of the cells using a confocal microscope. The results are as follows: Figure 4 , the level of molecularly induced autophagy can be preliminarily judged by GFP-LC3 fluorescence.

[0084] When the cells were only treated with the carrier molecul...

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Abstract

The invention relates to a medicine as well as a preparation method and application thereof and in particular relates to a medicine with an antigen processing improving function as well as a preparation method and application thereof. The medicine is prepared from a cell autophagy induction molecule, a tumor antigen molecule and a carrier, wherein the autophagy induction molecule and the tumor antigen molecule are connected onto the carrier. According to the medicine, the cell autophagy induction molecule and the tumor antigen molecule are connected onto the same carrier and have the synergistic interaction effect; compared with the effect of a manner of independently connecting the molecules with the carrier and entering cells respectively, the effect of the two molecules is remarkably enhanced; autophagy is effectively induced so that antigen presentation is enhanced; finally, the aim of improving the effect of treating tumors is realized; moreover, autophagy is enhanced by the medicine so that the antigen presentation is improved and the medicine provides a new thought for development of tumor immunotherapy.

Description

technical field [0001] The invention belongs to the field of tumor immunotherapy and relates to a medicine and its preparation method and application, in particular to a medicine with the function of improving antigen processing and its preparation method and application. Background technique [0002] In recent years, tumor immunotherapy has attracted much attention because of its significant curative effect on conventionally ineffective advanced tumors. It specifically eliminates cancerous cells by activating immune cells in the body. This treatment method has the advantages of strong specificity, long duration of action and few side effects, and has always been considered as the ultimate means of curing tumors. After Tumor Immunotherapy was rated as the top ten technological breakthroughs of the year by Science in 2013, the development of Tumor Immunotherapy has received more attention. [0003] Tumor immunotherapy is a new tumor treatment method following surgery, drug t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/59A61K38/10A61K38/08A61K39/385A61P35/00C08G18/28C08G18/50
CPCA61K38/08A61K38/10A61K39/385A61K2039/62C08G18/2865C08G18/5021A61K2300/00
Inventor 王浩王羿林耀新
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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