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Cell membrane coated nano bait for removing RANKL, preparation and application thereof

A cell membrane and nanotechnology, applied in the field of biomaterials and medicine, can solve problems such as complex manufacturing process, unsatisfactory biological distribution, and limited application of antibody resistance

Pending Publication Date: 2022-01-14
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, existing RANKL inhibitors (mainly protein drugs) often have limited applications due to shortcomings such as short blood circulation time, suboptimal biodistribution, complex manufacturing process, and antibody resistance.

Method used

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  • Cell membrane coated nano bait for removing RANKL, preparation and application thereof
  • Cell membrane coated nano bait for removing RANKL, preparation and application thereof
  • Cell membrane coated nano bait for removing RANKL, preparation and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0065] The preparation method of nano-bait

[0066] The application also provides a method for preparing the nanobait of the application, the method comprising:

[0067] (A) providing nanocores;

[0068] (B) providing cell membranes of osteoclast precursor cells;

[0069] (C) wrapping the cell membrane on the nano-core to form the nano-bait.

[0070] The nano-cores of the present application can be prepared from raw materials by various methods known in the art (such as using nano-precipitation method), and can also be purchased directly from various suppliers. The nano-core can have an opposite potential to the cell membrane to form a charge attraction to further stabilize the nano-bait.

[0071] Osteoclast precursor cell membranes can be obtained by cell lysis and separation, for example, the lysis includes: ultrasonic lysis, enzymatic lysis, chemical lysis, homogenate lysis and / or hypotonic swelling lysis; the separation includes: centrifugation (such as stepwise Cent...

Embodiment

[0104] The present application will be further elaborated below in conjunction with specific embodiments. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. Those skilled in the art can make appropriate modifications and changes to this application, and these modifications and changes are all within the scope of this application.

[0105] For the experimental methods that do not indicate specific conditions in the following examples, conventional methods in the art can be used, for example, with reference to "Molecular Cloning Experiment Guide" (third edition, New York, Cold Spring Harbor Laboratory Press, New York: Cold Spring Harbor Laboratory Press, 1989), "Animal Cell Culture" (Animal Cell Culture, edited by R.I. Freshney, 1987) or according to the conditions suggested by the supplier. The DNA sequencing method is a routine method in the art, and commercial com...

Embodiment 1

[0108] Embodiment 1, preparation and characterization of RAW-PLGA nano decoy (nanodecoy)

[0109] according to figure 1The procedure described in A prepares the cell membrane-coated nanocomposite of the present application——RAW-PLGA nanobait. Concrete preparation steps are as follows:

[0110] (1) Preparation of membrane material: mouse monocyte / macrophage-like cells RAW 264.7 (purchased from the Cell Bank of the Chinese Academy of Sciences, catalog number SCSP-5036, the medium is DMEM containing 10% FBS, 37 ° C, 5% CO 2 ) suspended in 20mM Tris·HCl (pH 7.5), 10mM KCl, 75mM sucrose, 2mM MgCl 2 and protease / phosphatase inhibitors (purchased from Pierce, Cat. No. A32953, each tablet dissolved in 10 mL of solution) in homogenization buffer. The cells in the suspension were disrupted with a JY 92-IIN homogenizer (75W), and then the supernatant was collected by centrifugation at 20000g for 25 minutes, and the cell membrane was collected by centrifugation of the supernatant at 1...

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Abstract

The invention provides a cell membrane coated nano bait for removing a nuclear factor kappa B receptor activation factor ligand (RANKL), preparation and application thereof. Specifically, the invention provides a nano bait, which comprises: (a) a nano core; and (b) a osteoclast precursor cell membrane coated with the nano core. The nano bait can be RAW-PLGA, wherein the RAW cell membrane is a cell membrane of osteoclast precursor cells, and PLGA is a nano core. The invention also provides a preparation method and application of the nano bait, especially application in treatment of osteoclast excess or hyperfunction related diseases (like osteoporosis). The nano bait provided by the invention can effectively remove highly expressed RANKL in osteoporosis, at the same time can escape from macrophage capture and has long-term blood circulation after systemic application, thus having great potential in clinical treatment of osteoporosis.

Description

technical field [0001] This application belongs to the field of biomaterials and medicine. Specifically, the present application relates to cell membrane coating technology, and more specifically relates to a nanocomplex coated with osteoclast membrane, which can effectively bind and clear highly expressed RANKL. Background technique [0002] Osteoporosis is a progressive bone disease characterized by loss of bone mineral density and quality, disruption of bone microarchitecture, and increased bone fragility. For women, postmenopausal osteoporosis caused by estrogen deficiency is the most common type, with approximately 50% of women experiencing at least one fracture after menopause. In postmenopausal women, estrogen deficiency leads to upregulation of RANKL, which further activates the nuclear factor-κB (NF-κB) pathway by binding to RANK in osteoclast precursor cells, thereby upregulating c-Fos gene expression. In addition, RANKL activates the mitogen-activated protein ki...

Claims

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Application Information

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IPC IPC(8): A61K9/16A61K47/02A61K47/34A61K47/36A61K47/42A61K35/26A61K35/28A61K35/32A61P1/02A61P19/08A61P19/10A61P35/00
CPCA61K9/1676A61K9/1647A61K9/1658A61K9/1652A61K9/1611A61K35/28A61K35/26A61K35/32A61P19/08A61P19/10A61P1/02A61P35/00
Inventor 殷黎晨周炀
Owner SUZHOU UNIV
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