Nanoparticle carrier containing TLR3 ligand in PRRs (pattern recognition receptors) as well as preparation method and application of nanoparticle carrier

A technology of pattern recognition receptors and nanoparticles, applied in the field of medicine, can solve the problems of lack of preventive or therapeutic vaccines, and achieve the effect of enhancing the production of interferon and pro-inflammatory factors

Active Publication Date: 2018-01-09
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are currently no effective preventive or therapeutic vaccines for many viral infections (such as HIV, hepatitis B virus, and hepatitis C virus infection)

Method used

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  • Nanoparticle carrier containing TLR3 ligand in PRRs (pattern recognition receptors) as well as preparation method and application of nanoparticle carrier
  • Nanoparticle carrier containing TLR3 ligand in PRRs (pattern recognition receptors) as well as preparation method and application of nanoparticle carrier
  • Nanoparticle carrier containing TLR3 ligand in PRRs (pattern recognition receptors) as well as preparation method and application of nanoparticle carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The present embodiment provides a nanoparticle carrier comprising a TLR3 ligand in a pattern recognition receptor, the nanoparticle carrier uses calcium phosphate nanoparticles as a core, the surface of the core is grafted with PEI, and the surface of the core is adsorbed and loaded with Poly(I: C), the outer core is a silica shell, and the silica of the shell is covalently bonded with thiol groups or amino groups.

[0047] The preparation method of the nanoparticle carrier comprising the TLR3 ligand in the pattern recognition receptor includes the following steps:

[0048] The nanoparticles were prepared using the following chemicals: branched polyethylene (PEI), calcium lactate, diammonium phosphate, tetraethoxysilane (TEOS), trimethoxysilane (MPS), triethoxysilane (APTES), cyclohexane-1-carboxylic acid-3-sulfosuccinimide ester (sulfo-SMCC); , ammonia, cross-linked dextran.

[0049] Step 1: Synthesis of stable PEI-calcium phosphate nanoparticles (CaP / PEI)

[0050] I...

Embodiment 2

[0061] The present embodiment provides a nanoparticle carrier comprising a TLR3 ligand in a pattern recognition receptor, the nanoparticle carrier uses calcium phosphate nanoparticles as a core, the surface of the core is grafted with PEI, and the surface of the core is adsorbed and loaded with Poly(I: C), the outer core is a silica shell, and the silica of the shell is covalently bonded with thiol groups.

[0062] The preparation method of the above-mentioned nanoparticle carrier containing the TLR3 ligand in the pattern recognition receptor is basically the same as that of Example 1, the difference is that:

[0063] Step 2: Synthesis of poly(I:C)-loaded CaP / PEI (CaP / PEI / poly(I:C))

[0064] 1 mL of CaP / PEI was mixed with 100 μL of water-soluble poly(I:C)-Cy5 containing fluorescein, and stirred at room temperature for 30 minutes.

[0065] The poly(I:C)-Cy5 used in this example can also be replaced by poly(I:C)-TRITC. Embodiment three

Embodiment 3

[0066] The present embodiment is to CaP / PEI-Cy5 / Poly(I:C) / SiO 2 The cytotoxicity of -SH was tested, and the specific steps included:

[0067] 1.1 Experimental materials:

[0068] 6-well plates were purchased from Thermo NUNC, Denmark

[0069] RPMI1640 (high glucose), fetal bovine serum, penicillin and streptomycin were purchased from Gibco, USA.

[0070] MTT cell proliferation and cytotoxicity kits were purchased from China Biyuntian Company.

[0071] 1.2 Experimental method:

[0072] 1.2.1 Cell culture

[0073] THP-1 cells were mixed with different concentrations of CaP / PEI-FITC / Poly(I:C) / SiO 2 -SH nanoparticles were cultured for 1h / 24h.

[0074] 1.2.2 MTT cytotoxicity test

[0075] Add 10ul of MTT solution (5mg / ml prepared in PBS, pH=7.4) to each well. Continue to incubate for 4h, terminate the culture, and carefully aspirate and discard the culture supernatant in the well. Add 100ul DMSO to each well and shake for 10min to fully dissolve the crystals. Select a wave...

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Abstract

The invention provides a nanoparticle carrier CaP / PEI / poly(I:C) / SiO2-SH containing a TLR3 ligand in PRRs (pattern recognition receptors). The compound is characterized in that calcium phosphate nanoparticles are taken as a core, the surface of the core is grafted with PEI, Poly(I:C) is adsorbed and supported on the surface of the core, a SiO2 shell is located outside the core, and under the actionof trimethoxysilane, the SiO2 shell can carry thio groups to be functionalized and can carry different fluorescent dyes such as FITC, Cy5 and the like. Further, the invention provides a preparation method and a medical application of the compound. Nanoparticles are stable in property and lower in cytotoxicity and can carry different fluorescent dyes, binding of poly(I:C) and TLR3 can be realizedeffectively with the nanoparticles as the carrier, production of interferons and proinflammatory factors is enhanced by activating a TLR3 mediated signaling pathway, and the nanoparticles are expectedto be used as drug in-vivo imaging tools and vaccine carriers.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a nanoparticle carrier containing a TLR3 ligand in a pattern recognition receptor and a preparation method and application thereof. Background technique [0002] Among various nanoparticles, calcium phosphate nanoparticles have the characteristics of high biocompatibility, high biodegradability, small size and high affinity, and can be covalently functionalized with some nucleic acids, because of their considerable application prospects. The nanoparticles have been used in various fields, such as transfection, gene silencing, drug delivery, photodynamic therapy, etc. Some macromolecules are often unable to penetrate the cell membrane, therefore, an effective carrier such as nanoparticles is required. [0003] Calcium phosphate nanoparticles, about 100 nm in size, are easily taken up by cells and subsequently dissolved in lysosomes. Multiple studies have demonstrated that multi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/58A61K47/69A61K31/713A61K49/00A61P31/12G01N33/551
Inventor 吴珺邹诗黄顺梅杜艳芹马赛厄斯·埃普尔维多利亚·索科洛娃
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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