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A kind of natural biological nano-targeted drug complex material and its preparation method and application

A bio-nano-composite technology, applied in the field of medical materials, can solve the problems of yet to be elucidated interaction laws and effects, lack of functional nano-carrier materials, and difficulty in large-scale industrial production. Good compatibility, size controllable effect

Active Publication Date: 2021-06-08
ZHUKANG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is a basic research issue. After nanoparticles enter the human body, their interactions with human organs, tissues, cells, and molecules, especially the rules and effects of interactions with cells have yet to be elucidated.
[0006] 2) Lack of functional nanocarrier materials for intravascular use
At present, except for albumin, phospholipids and their analogs, and PEGylated polylactic acid, there are no other synthetic polymer materials suitable for intravascular use. The main reason is the lack of complete safety evaluation data.
[0007] 3) Production cost and process issues
At present, carrier materials such as PLGA with relatively good performance and little side effects on the human body have to be processed by complex chemical methods, the production cost is high, and the output is low, so they cannot be widely used in clinical practice.
[0008] 4) The quality evaluation system of the nano drug delivery system needs to be established, and its industrial engineering technology needs to be developed
Because chitosan, sodium alginate, hyaluronic acid, etc. have to go through cumbersome chemical processing or use microbial fermentation, the production cost is high, and it is difficult to implement large-scale industrial production
However, protein products are mainly derived from human blood, so the source is limited, and the large-scale preparation of carriers is subject to great constraints. The production process is complicated, the production cost is high, and allergic reactions are prone to occur.

Method used

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  • A kind of natural biological nano-targeted drug complex material and its preparation method and application
  • A kind of natural biological nano-targeted drug complex material and its preparation method and application
  • A kind of natural biological nano-targeted drug complex material and its preparation method and application

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preparation example Construction

[0041] The embodiment of the present invention also provides a preparation method of a natural biological nano-targeted drug complex material, comprising the following steps:

[0042] S1. Take 200 μL of Bacillus amyloliquefaciens strain stored at -20°C with 50% glycerol, add it to sterilized 5mL LB liquid medium, place it in a temperature-controlled shaker at 37°C at 180rpm and culture overnight to complete the activation of the strain;

[0043] S2. Take 200 μL of the activated bacterial solution and add it to the cooled LB solid medium plate. After spreading it evenly with a glass rod, place the culture plate upside down in a constant temperature incubator at 37°C and cultivate it for 7 days. Bacillus amyloliquefaciens begins to grow and transform into spores; collect the spores of Bacillus amyloliquefaciens grown on the plate, wash the spore suspension 3 times with double distilled water, and ultrasonicate 5 times under ice bath conditions, each time for 3 minutes, and each u...

Embodiment 1

[0050] In this example, the multifunctional targeting natural bionano drug complex is prepared by the following method, which specifically includes the following steps:

[0051] Step 1. Using the carbodiimide method, redissolve the prepared BA (Bacillus amyloliquefaciens cell wall peptidoglycan) (6 million units) in 1 mL of MES buffer (0.1 mol / mL, pH=5.5), Add excess EDC and NHS to it successively (the molar ratio of substances is 1.5:1); place it on a shaker and activate it at 4° C. for 90 minutes to activate free carboxyl groups.

[0052] Step 2, wash with double distilled water, centrifuge (12000rpm, 3min), repeat 3 times. Redissolve the activated spores in 1ml of PBS buffer (0.1mol / L, pH=7.2), add 20,000 units of UK to it, place it on a shaker, react at room temperature for 20 minutes, then add 0.05mg of RGD, and activate at room temperature For 100 minutes, the primary amino group was reacted with the activated carboxyl group to form an amide bond linking UK, RGD and BA....

Embodiment 2

[0057] In this embodiment, the evaluation of the in vitro thrombolytic ability of the multifunctional targeted natural bionano drug complex (targeted thrombolytic drug) specifically includes the following steps:

[0058] Step 1. Select 300-350g of healthy Wistar rat heart to collect blood, and bathe in 37°C water for 2 hours to inactivate plasminogen and form cellulose thrombus.

[0059] Step 2. Divide the formed cellulose thrombus into thrombi with a weight of about 0.1g and uniform shape, and divide them randomly into five groups: PBS group, high-dose UK group, low-dose UK group, high-dose UK-BA-RGDs group, Low-dose UK-BA-RGDs group. 10 in each group, filter paper to dry the surface moisture, mark 1 to 10, weigh and record.

[0060] Step 3. Add 1ml of PBS, 2000IU UK, 5000IU UK, 2000IU UK-BA-RGDs, 5000IU UK-BA-RGDs according to the corresponding groups and incubate in a 37°C water bath for 2 hours to fully react.

[0061] Step 4. Take out the incubated emboli, blot the surf...

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Abstract

The invention discloses a natural biological nano-targeted drug composite material and its preparation method and application. The composite material includes a carrier and an active ingredient loaded on the carrier, and the carrier and the active ingredient are connected by a covalent bond to form a Complex, the size of the complex is 1-1000 nanometers, the carrier is cell wall peptidoglycan, the size is 1-1000 nanometers; 2 Or ‑OH or ‑COOH drugs and ‑NH-containing drugs with affinity for the lesion area 2 or ‑OH or ‑COOH targeting substances. The material obtained by the invention has good applicability, high safety, small particle size, uniform particle size, controllable size, long circulation effect in the body, good biocompatibility, no cytotoxicity, and immune activation effect , can improve the body's immunity. It can be loaded with a variety of drugs and targeted substances, so as to achieve the purpose of precise treatment.

Description

technical field [0001] The invention relates to the field of medical materials, in particular to a natural biological nano-targeted drug complex material and its preparation method and application. Background technique [0002] Targeted drug delivery system (TDDS) refers to a new drug delivery system that can selectively concentrate drugs in target tissues, target cells or specific organelles in cells. It is the fourth stage in the development of pharmaceutical dosage forms, mainly for the treatment of diseases such as tumors. Targeted drug delivery methods include carrier-mediated, receptor-mediated, prodrug, and chemical delivery systems. Among them, carrier-mediated is the most commonly used method, which can be divided into liposomes, nanoparticles, microspheres, etc. according to different carriers. Mainly through passive and active targeted drug delivery. The advantages of targeted drug delivery are obvious. It can concentrate the drug in the target area and leave l...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/61C08B37/00C12N3/00C12N13/00C12R1/07
CPCC08B37/00C08B37/0003C12N3/00C12N13/00
Inventor 齐锦生胡涌刚巩文建高雪涵栗彦宁刘昆毛若男苗大壮
Owner ZHUKANG BIOTECHNOLOGY CO LTD
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