A multi-purpose polylysine fluorescent self-assembled nano-microsphere carrier and its preparation method and application

A technology of polylysine and nano microspheres, which is applied in the field of biomedicine to achieve good colloidal properties, avoid interference, and the effect of simple and easy preparation process

Active Publication Date: 2017-01-18
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it has not been seen that polylysine itself is used as the backbone. On the one hand, fluorescent molecules (rhodamine) are covalently linked through amide bonds, and nanoparticles are prepared by self-assembly technology. On the other hand, drug molecules can be released by covalently combining with click chemistry technology. At the same time, it can perform in vivo and in vitro tracer imaging analysis of modified drugs, target location and target protein capture and separation, and report on polylysine nanomaterials that integrate the above functions and uses

Method used

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  • A multi-purpose polylysine fluorescent self-assembled nano-microsphere carrier and its preparation method and application
  • A multi-purpose polylysine fluorescent self-assembled nano-microsphere carrier and its preparation method and application
  • A multi-purpose polylysine fluorescent self-assembled nano-microsphere carrier and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Preparation and azide modification of highly fluorescent polylysine modified nanospheres

[0035] Dissolve 5 g of polylysine hydrochloride in 500 mL of pure water and load it on activated 001×7 cation exchange resin (2×80 cm) for desalting at a rate of 1 mL / min. Equilibrate with 1500 mL of pure water until the effluent is neutral, elute with 5% ammonia water, collect the eluate, distill under reduced pressure, concentrate and dry to obtain 1.9 g of white to light yellow solid.

[0036] Take 200 mg of the above-mentioned desalted polylysine in 10 mL of pure water, stir to dissolve completely to obtain an aqueous phase solution; at the same time, mix 13 mL of gasoline, 12 mL of carbon tetrachloride and 1.25 mL of Span-80, and stir The organic phase is obtained homogeneously. Add 20 mg of NHS-Rhodamine (NHS-Rhodamine, Pierce 46406) to the organic phase, mix well, and set aside. Quickly pour the above water phase into the organic phase, stir at 12,000 rpm for 3...

Embodiment 2

[0038] Example 2, Preparation and Azide Modification of Polylysine Fluorescent Nanospheres

[0039] Dissolve 50 mg of the desalted polylysine in Example 1 above in 10 mL of pure water to obtain an aqueous phase solution; simultaneously mix 5 mL of gasoline, 15 mL of carbon tetrachloride and 0.5 mL of Span-80, and stir evenly to obtain an organic Mutually. Add 0.1 mg of NHS-Rhodamine (NHS-Rhodamine, Pierce 46406) to the organic phase, stir to dissolve, and set aside for later use. Pour the above water phase into the organic phase quickly, stir at 20,000 rpm for 15 seconds at high speed, repeat 3 times to obtain a uniform emulsion, and transfer it to a separatory funnel. Add 20 mL of petroleum ether, stir for 5 minutes, then add 10 mL of pure water, continue stirring for 5 minutes, let stand to separate layers, collect the aqueous phase, and obtain crude fluorescent nanospheres. The crude product was centrifuged at 12,000 rpm for 10 minutes, the precipitate was discarded, and ...

Embodiment 3

[0040] Example 3. Preparation and highly azide modification of polylysine fluorescent nanospheres

[0041] Dissolve 20 mg of the desalted polylysine in Example 1 above in 10 mL of pure water to obtain an aqueous phase solution; at the same time, mix 15 mL of gasoline, 5 mL of carbon tetrachloride and 0.2 mL of Span-80, and stir evenly The organic phase is obtained. Add 2 mg of NHS-Rhodamine (NHS-Rhodamine, Pierce 46406) (0.002 mmoL) to the organic phase, stir to dissolve, and set aside for later use. Pour the above water phase into the organic phase quickly, stir at 10,000 rpm for 20 seconds at high speed, repeat 3 times to obtain a uniform emulsion, and transfer it to a separatory funnel. Add 10 mL of petroleum ether, stir for 5 minutes, then add 10 mL of pure water, continue stirring for 5 minutes, let stand to separate layers, collect the aqueous phase, and obtain crude fluorescent nanospheres. The crude product was centrifuged at 12,000 rpm for 10 minutes, the precipitat...

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Abstract

The invention discloses a multipurpose polylysine fluorescence self-assembly nano microsphere carrier and a preparation method and application thereof. By nano self-assembly technique, rhodamine marked polylysine fluorescence self-assembly nano microspheres can be prepared, azide group side chains with disulfide bonds can be modified, and by use of a click chemistry method, an alkynyl modified drug is connected with the fluorescent nano microspheres. The polylysine self-assembly nano microspheres linked with the drug can be used for tracing imaging analysis of the modified drug in animal and cell level. At the same time, the polylysine self-assembly nano microspheres can also be used to capture binding proteins of the modified drug, can release target binding proteins and associated proteins by cleavage of the disulfide bonds, and can be used for the study of drug effect targets. The invention provides a simple and fast tool and method for the study of the drug effect targets, and the method integrates drug tracing, target positioning and target protein capture and separation functions, is simple and practical, and has good application prospect in the study of drug action mechanisms and target discovery.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a preparation process of a polylysine nanometer self-assembled microsphere carrier connected with a fluorescent tracer group, and a method for covalently coupling modified drugs by click chemistry technology. And using the drug-loaded microspheres prepared by the present invention, the whole animal and cell-level tracer imaging analysis of the modified drug, drug target positioning, and application of target protein capture and separation. Background technique [0002] ε-Poly-L-lysine (Poly-L-lysine, PL) is a linear polymer of the homotype monomer formed by sequentially linking 25 to 30 lysine residues through amide bonds. Polylysine is white or light yellow powder, strong hygroscopicity, slightly bitter taste, not affected by pH value. ε-polylysine is a polypeptide with antibacterial effect. It has the advantages of broad antibacterial spectrum, high water solubility, good th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/16A61K47/48A61K49/00B82Y5/00C07K1/34
Inventor 白钢崔庆新侯媛媛姜民
Owner NANKAI UNIV
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