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Targeted adhesion chitosan material and application thereof

A chitosan and targeting technology, applied in medical preparations of non-active ingredients, gene therapy, anti-tumor drugs, etc., can solve problems affecting gene capacity, etc., to reduce chemical toxicity, reduce steric hindrance, and function enhanced effect

Active Publication Date: 2010-06-23
ZHANGJIAGANG IND TECH RES INST CO LTD DALIAN INST OF CHEM PHYSICS CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the amino group is usually more reactive than the hydroxyl group, and is often used as a grafting reaction site. At the expense of some amino groups on the chitosan molecular chain, the unique biological activity of chitosan is sacrificed, which also affects its gene capacity as carrier

Method used

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  • Targeted adhesion chitosan material and application thereof
  • Targeted adhesion chitosan material and application thereof
  • Targeted adhesion chitosan material and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0046] 1. At room temperature, 0.1g chitosan (Mw=1500kDa, DD=40%) was dissolved in 100mL of aqueous acetic acid with a molar concentration of 0.05M, and the pH of the solution was adjusted to pH5.0 to form a transparent 0.1% (w / v) chitosan solution;

[0047] 2. Take the photosensitive cross-linking agent SANPAH (according to the ratio of the amount of substances of the photosensitive cross-linking agent and the glucosamine residue in chitosan is 1: 1 metering), join in the ethanol, then add in the photosensitive cross-linking agent solution The ratio of SANPAH to RGD cyclic peptide is 1:2 in RGD cyclic peptide aqueous solution, and the final concentration of photosensitive cross-linking agent is 10 μM. React in the dark for 90 minutes at room temperature to form a mixture of RGD cyclic peptide-SANPAH and unreacted raw materials;

[0048] 3. According to the amount of 1: 1 in the ratio of the amount of substances of glucosamine residues in the photosensitive cross-linking age...

preparation example 2

[0052] 1. At room temperature, dissolve 4g of chitosan (Mw=10kDa, DD=60%) in 100mL of acetic acid aqueous solution with a molar concentration of 0.1M, and adjust the pH of the solution to pH5.5 to form a transparent 4% (w / v ) chitosan solution;

[0053] 2. Take the photosensitive cross-linking agent SANPAH (according to the ratio of the amount of substances of the photosensitive cross-linking agent and the glucosamine residue in chitosan is 1: 6 metering), join in the ethanol, then add in the photosensitive cross-linking agent solution The ratio of SANPAH to GRGD is 1:20 in GRGD aqueous solution, and the final concentration of photosensitive cross-linking agent is 0.1 mM. React in the dark for 120 minutes at room temperature to form a mixture of GRGD-SANPAH and unreacted raw materials;

[0054] 3. Add chitosan solution to GRGD-SANPAH and its raw material mixture according to the ratio of photosensitive cross-linking agent and glucosamine residue in chitosan as 1:6, mix well, ...

preparation example 3

[0058] 1. At room temperature, dissolve 1 g of chitosan (Mw=148kDa, DD=80%) in 100 mL of aqueous acetic acid with a molar concentration of 0.05 M, adjust the pH of the solution to pH 4.5, and form a transparent 1% (w / v ) chitosan solution;

[0059] 2. The ratio of sulfo-SANPAH and GRGDY is 1:1, respectively added to water, the final concentration of photosensitive cross-linking agent is 2M, GRGDY and sulfo-SANPAH are reacted at room temperature in the dark for 120min to form GRGDY-SANPAH and its untreated The mixed solution of raw materials for the reaction;

[0060] 3. Add chitosan solution to GRGDY-SANPAH and its raw material mixture according to the ratio of photosensitive cross-linking agent and glucosamine residue in chitosan as 1:2, mix well, and then UV light 15min.

[0061] 4. Add the above reaction mixture solution into a dialysis bag with a molecular weight cut-off=14kDa, and dialyze until the dialyzed aqueous solution has no detectable change in ultraviolet absorp...

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Abstract

The invention relates to a targeted adhesion chitosan material and application thereof. Under the action of dual-functional (N-hydroxysuccinimide / azide) light-sensitive cross-linking agent, through grafting reaction by UV photochemical activation, cell adhesion peptide is grafted on the chitosan to obtain the safe (non-toxic and degradable) targeted adhesion chitosan material which is characterized in that the biocompatibility is good, the biological function is good and the dissolubility under faintly acid condition is obviously enhanced. Through self assembling method, the targeted adhesion chitosan material can form stable nano particles with grain size of 100mm-500mm and surface positive charge of 10mV-30mV and the stable nano particles can be used as the carriers of DNA (20bp-10kb) or RNA [dsRNA (500-700bp), small RNA (siRNA, microRNA or shRNA)] and other nucleic acid drugs. When the carried DNA or RNA is protected from being degraded by the corresponding enzyme, the carried DNA or RNA can target at the overexpressed integrin receptors on the surface of tumor cells, so cell transfection and release in cells are facilitated to cure tumors.

Description

technical field [0001] The invention relates to a nucleic acid drug for tumor treatment. Specifically, a natural polysaccharide material with targeted adhesion is used to prepare a nano-carrier, which can effectively protect and target the nucleic acid drug into cells and realize release. Background technique [0002] Gene therapy has promising applications in replacing dysfunctional genes and in tumor therapy. Gene therapy has three basic elements: target gene, expression vector and delivery vector. RNA interference (RNAinterference) mediated by RNA [dsRNA or small RNA (siRNA, microRNA or shRNA)] is to specifically bind to the messenger RNA (mRNA) complementary to its sequence, and degrade the mRNA by nuclease to block its coding The expression of protein can realize gene silencing, so it has important scientific significance and application value in the field of treatment of cancer, influenza, hepatitis and other viral infection diseases. However, naked small RNA has poo...

Claims

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Application Information

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IPC IPC(8): C08B37/08A61K47/48A61K48/00A61P35/00A61K47/61
Inventor 马小军杨艳刘袖洞于炜婷
Owner ZHANGJIAGANG IND TECH RES INST CO LTD DALIAN INST OF CHEM PHYSICS CHINESE ACADEMY OF SCI
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