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Nano-drug carrier, and preparation method and application thereof

A technology for drugs and solid substances, applied in the field of biomedicine, can solve the problems of many defective proteins, severe encapsulation methods, limited types of encapsulated drugs, etc., and achieve a good effect of sustained release.

Pending Publication Date: 2021-02-09
KUNSHAN XINYUNDA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to overcome the problems existing in the existing ferritin entrapment methods, such as severe entrapment method, many defective proteins after entrapment, limited types of entrapped drugs, and changes in drug activity after entrapment.

Method used

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  • Nano-drug carrier, and preparation method and application thereof
  • Nano-drug carrier, and preparation method and application thereof
  • Nano-drug carrier, and preparation method and application thereof

Examples

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preparation example Construction

[0055] In one aspect, the invention provides a kind of preparation method of ammonium salt-cage protein, comprises the steps:

[0056] Step 1: preparing a solution containing caged protein and ammonium salt;

[0057] Step 2: preparing a solution of clathrate protein with ammonium salt in the inner cavity; and

[0058] Optionally, step 3 is also included: removing free ammonium salts outside the cage protein cage.

[0059] In a specific embodiment, wherein, in the solution described in step 1, the concentration of the clathrate protein is 0.5-20 mg / mL, preferably 2-15 mg / mL, more preferably 4-12 mg / mL, further preferably 5-10 mg / mL;

[0060] And / or the ammonium salt concentration is 1-4000mmol / L, preferably 1-2500mmol / L, preferably 10-500mmol / L, more preferably 50-500mmol / L, further preferably 30-300mmol / L, even more preferably 40- 200mmol / L, such as 40-100mmol / L, 150-350mmol / L, 150-250mmol / L, 200mmol / L.

[0061] In one aspect, the invention provides a kind of preparation ...

Embodiment 1

[0109] Example 1 prepares inner chamber entrapment (NH 4 ) 2 SO 4 HFn

[0110] In this example, lumen entrapped (NH 4 ) 2 SO 4 Ferritin 1#.

[0111] 1. Replace the solution outside the cage with (NH 4 ) 2 SO 4 Solution: pH 8.0, HFn solution (protein concentration 13.2 mg / ml) prepared by 50 mM Tris-HCl was used in an ultrafiltration centrifuge tube with a molecular weight cut-off of 100 KDa, and the 4 ) 2 SO 4 The aqueous solution was ultrafiltered and centrifuged 3 times (3700rpm each time, 20min) to obtain (NH 4 ) 2 SO 4 and HFn solution. Then add 200mmol / L(NH 4 ) 2 SO 4 Dilute with aqueous solution, adjust the protein concentration to 10mg / mL, place at 60°C for 4h.

[0112] 2. Remove the cage solution (NH 4 ) 2 SO 4 : use ddH 2 O was replaced by ultrafiltration centrifugation for 3 times, 12000rpm each time, 20min each time, the molecular weight cut-off of the ultrafiltration centrifuge tube was 3KDa, the final filtrate was clear and colorless, and the ...

Embodiment 2

[0113] Example 2 prepares inner chamber entrapment (NH 4 ) 2 SO 4 HFn

[0114]In this example, the inner cavity entrapped (NH 4 ) 2 SO 4 Ferritin 2#.

[0115] 1. Replace the solution outside the cage with (NH 4 ) 2 SO 4 Solution: in On the 150 system, use a Superdex75pg packing self-packing column (XK 50 / 20, 10cm) to carry out desalination and liquid replacement treatment on the sample, and the buffer solution is 200mmol / L (NH 4 ) 2 SO 4 solution. Specifically, 6 mL of HFn solution (protein concentration 16.5 mg / ml) prepared with pH 8.0 and 50 mM Tris-HCl was loaded onto a self-packed column, and 200 mmol / L (NH 4 ) 2 SO 4 Solution elution, collect the eluent of HFn under the condition of A280 ultraviolet absorption, get containing (NH 4 ) 2 SO 4 and HFn solution. Then add 200mmol / L (NH 4 ) 2 SO 4 The solution was diluted to a protein concentration of 4 mg / mL, and placed at 60°C for 4 hours.

[0116] 2. Remove (NH in the solution outside the cage 4 ) 2...

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Abstract

The invention discloses a nano-drug carrier, and a preparation method and application thereof. The nano-drug carrier disclosed by the invention is an ammonium salt-cage protein; ammonium salt is entrapped in an inner cavity of a cage protein; and the ammonium salt-cage protein is prepared by the following steps: step 1, preparing a solution containing the cage protein and the ammonium salt; step 2, preparing a solution of the cage protein with the inner cavity carrying the ammonium salt, and optionally, step 3, removing free ammonium salt in the solution outside a cage protein cage. The process for preparing the ammonium salt-cage protein by the preparation method disclosed by the invention is mild; denaturation and defect of the cage protein cannot be caused; particles are uniform and consistent; and furthermore, the inner cavity of the ammonium salt-cage protein is used for entrapping a drug, so that the drug-loaded ammonium salt-cage protein with an ideal drug entrapping effect anda better slow release effect can be prepared.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular to an ammonium salt-cage protein and its preparation method and application, and an ammonium salt-cage protein-drug formed by contacting the ammonium salt-cage protein with a drug and its preparation method and application. Background technique [0002] Ferritin is a kind of natural iron storage protein that widely exists in animals, plants and microorganisms. Due to its unique spherical shell structure and reversible self-assembly properties, it can be used as a carrier of various drugs. Ferritin is composed of 24 subunits, each two subunits form a group of antiparallel, forming an approximate regular octahedron, forming a hollow spherical molecule with 4-3-2 heavy axis symmetry, and each ferritin molecule forms 12 binary Heavy axis channels, 8 triple axis channels and 6 quad axis channels. Utilizing the reversible self-assembly property of denaturation and renaturation of fer...

Claims

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Application Information

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IPC IPC(8): A61K47/69A61K9/51A61K9/52A61K47/42A61K31/704A61P35/00
CPCA61K47/6949A61K9/5169A61K31/704A61P35/00
Inventor 柯天一温佰刚李彦良檀琳劳芳欧阳芳幸徐菊芳
Owner KUNSHAN XINYUNDA BIOTECH CO LTD
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