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Nano preparation for combined transportation of nucleic acids and polypeptides and preparation method

A nano-preparation, a variety of polypeptide technology, applied in powder delivery, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc. loop effect

Active Publication Date: 2016-02-17
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few studies on the carriers of peptide drugs. In fact, peptide drugs have small molecular weight and clear targets, and play an important role in blocking or regulating diseased genes. See literature [Ellerby, H.M et al. Antitumor activity of peptides, recorded in Natural Medicine, 1999,5,1032-8]

Method used

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  • Nano preparation for combined transportation of nucleic acids and polypeptides and preparation method
  • Nano preparation for combined transportation of nucleic acids and polypeptides and preparation method
  • Nano preparation for combined transportation of nucleic acids and polypeptides and preparation method

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preparation example Construction

[0042] The embodiment of the present invention relates to a method for preparing nanoparticles co-delivered with polypeptides and nucleic acids, comprising the following steps:

[0043] a. Preparation of cationic liposomes by film dispersion method: In this experiment, cationic lipid (2,3-dioleoxypropyl)trimethylammonium chloride (DOTAP) and neutral lipid 1,2-diole Acyl-sn-glycero-3-phosphoethanolamine (DOPE) in chloroform with a mass ratio of 1:1 was rotovapped to remove the solvent, then hydrated overnight and sonicated for 30-60 minutes.

[0044] b. Using a degradable bond to connect the GGWYRQV sequence as a polypeptide drug with polyarginine (R11) through different sequence ends to construct Pa and Pb two polypeptides that can continuously degrade and release the polypeptide drug GGWYRQV.

[0045]c. Preparation of nano-preparation: use the liposome prepared above to dilute to a certain concentration (0.04mg / mL), and mix it with a certain concentration of polypeptide (0.16...

Embodiment 1 4

[0046] The preparation method of embodiment 1 quaternary nanoparticles

[0047] Such as figure 1 As shown, the system for measuring the physicochemical properties of quaternary nanoparticles was carried out in aqueous solution. After mixing a certain amount of cationic liposomes and polypeptides, add them to the nucleic acid aqueous solution and continue to mix evenly. Incubate at room temperature for 30 minutes to prepare ternary nanoparticles, then add hyaluronic acid into the mixture and mix thoroughly, and let stand for 15 minutes to prepare quaternary nanoparticles. For the carrier system used in cell experiments, serum-free medium can be used instead of ultrapure water.

Embodiment 2 4

[0048] The gel electrophoresis of embodiment 2 quaternary nanoparticles

[0049] Weigh 0.3g of agarose powder, pour it into a conical flask, add 30ml of 1×TAE buffer solution into the conical flask, fully dissolve it under heating conditions, and add Gelredinwarter 3μL after the temperature drops to 65°C, Quickly pour it into the gel-making tank, insert the sample comb, and place it at room temperature for 0.5-1h until the gel becomes solidified. Then pull out the sample comb, and then pour TAE buffer into the sample tank to submerge the gel. Prepare 10 μL of quaternary nanoparticles in different proportions, mix 2 μL of 6× loading buffer with quaternary nanoparticles, load 10 μL of the sample, and load 10 μL of the nakedPGL3 plasmid as a control. Then electrophoresis was performed at a voltage of 110 mV for 45 minutes, the electrophoresis was turned off, the gel was taken out and placed in an ultraviolet gel imaging system for observation and electrophoresis images were re...

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Abstract

The invention aims to overcome defects that bio-macromolecular nucleic acid and polypeptide medicines degrade easily in bodies and can hardly reach target cells and provides a nano preparation for combined transportation of nucleic acids and polypeptides and a preparation method. According to the nano preparation, multifunctional cationic polypeptides and cationic liposomes / nucleic acid non-viral carriers which are safe to human bodies are adopted to coat polypeptide medicines and nucleic acid medicines simultaneously to form nanoparticles, and bio-macromolecular medicine carriers capable of treating diseases through combination of the polypeptides and the nucleic acids are prepared. The cationic liposomes and the polypeptides are mixed and then assembled with the nucleic acid medicines under the electrostatic effect, the nanoparticles with positive charges on surfaces are formed, and nano target particles with negative charges on surfaces are further formed under the electrostatic effect by covering hyaluronic acids with hydrogen bonds. The nano preparation has the advantages that the polypeptide medicines and the nucleic acid medicines are loaded simultaneously, the polypeptide medicines are released continuously through breakage of degradable bonds between the polypeptide medicines and poly-L-arginine, and the nucleic acids are released effectively through disassembly of the carriers.

Description

technical field [0001] The present invention relates to a drug delivery method in the technical field of medicine, in particular to a nano-preparation and preparation method for combined delivery of nucleic acid and polypeptide, especially a nano-preparation that can be used for two or more biomacromolecular drugs Combination biological pharmaceutical preparations for the treatment of diseases. Background technique [0002] Cancer is one of the major diseases that are difficult to cure. The current methods of chemotherapy, radiotherapy and surgery have disadvantages such as high toxicity, lack of targeting, great damage to normal tissues and cells, and unclean resection. The development of new small molecule drugs takes a long time and costs a lot. Biomacromolecular drugs (such as polypeptides, nucleic acids, and proteins) have specific therapeutic targets and low toxicity. In particular, nucleic acid and peptide drugs have clear structures and short preparation time. How...

Claims

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Application Information

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IPC IPC(8): A61K47/36A61K47/48A61K38/08A61K31/7088A61K9/14A61P35/00
Inventor 杜子秀徐宇虹梁瑶瑶
Owner SHANGHAI JIAO TONG UNIV
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