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Fusion protein SZ1 for targeted therapy of colorectal cancer caused by excessive activation of Rspo, and coupled nano-drug thereof

A fusion protein and over-activation technology, applied in the field of biomedicine, can solve the problems of ineffectiveness of colorectal cancer, and achieve the effect of increasing stability, good therapeutic effect and enhancing targeting.

Active Publication Date: 2021-08-27
NORTHEASTERN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the drug is not effective against colorectal cancer caused by other Rspo

Method used

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  • Fusion protein SZ1 for targeted therapy of colorectal cancer caused by excessive activation of Rspo, and coupled nano-drug thereof
  • Fusion protein SZ1 for targeted therapy of colorectal cancer caused by excessive activation of Rspo, and coupled nano-drug thereof
  • Fusion protein SZ1 for targeted therapy of colorectal cancer caused by excessive activation of Rspo, and coupled nano-drug thereof

Examples

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Effect test

Embodiment 1

[0057] A method for preparing the fusion protein SZ1 for targeted treatment of colorectal cancer caused by Rspo overactivation, specifically comprising:

[0058] In this embodiment, the expression vector of the fusion protein SZ1 for the targeted treatment of Rspo overactivated colorectal cancer is a plasmid expression vector, and the plasmid expression vector integrates the DNA sequence corresponding to the human FZD5 secretion signal peptide, V5 The DNA sequence corresponding to the tag peptide, the DNA sequence corresponding to the partial ectodomain of human LGR4, the DNA sequence corresponding to the part ectodomain of human ZNRF3, the DNA sequence corresponding to the amino acid of the linker domain connecting the two, and mouse IgG2α / human IgGH3 Fc The DNA sequence corresponding to the domain; the DNA sequence corresponding to the human FZD5 secretion signal peptide is the region from amino acid No. 1 to amino acid No. 26, the DNA sequence corresponding to the V5 tag pep...

Embodiment 2

[0103] The preparation method of gold nanorod SZ1-cRGD-AuNRs modified by SZ1 and RGD short peptide comprises the following steps:

[0104] Step 1: preparing gold nanorods;

[0105] ① Seed solution preparation: a certain volume of chloroauric acid solution (5mmol L -1 ) and an equal volume of CTABr (0.1mol·L -1 ) after mixing evenly, add sodium borohydride solution (0.5mL, 0.005mol·L -1 ), kept stirring at room temperature for 2 hours. ②Growth solution preparation: add 5mL 0.2mol·L to the reaction vessel successively - 1 CTABr, 5mL 1mmol L -1 HAuCl 4 Solution, 0.5mL5mmol·L -1 AgNO 3 Solution, 0.07mL 0.1mol L -1 Ascorbic acid solution, stirred for 3 minutes. ③Preparation of gold nanorods (Au NRs): Add 0.1mL ①seed solution to ② growth solution, stir for 2 minutes, and leave at room temperature to obtain fully grown Au NRs. like Figure 5 As shown, the prepared gold nanorods have significant absorption bands belonging to nano-gold at 755nm and 515nm, respectively, corr...

Embodiment 3

[0108] Silica-coated gold nanorods modified with SZ1 and RGD short peptide SZ1-cRGD-SiO 2 The preparation method of @AuNRs includes the following steps:

[0109] Step 1: Preparation of silica-coated gold nanorods: redisperse 50 mg of purified Au NRs in 25 mL of deionized water, add 50 μL of ammonia water (mass fraction 25%), and slowly add 1 mL of it to a concentration of 10 mmol L -1 tetraethyl orthosilicate / ethanol solution, and reacted at 40°C for 24 hours. After centrifuging to remove the supernatant, add ethanol to disperse, then centrifuge again to remove the supernatant, then disperse the obtained precipitate in water, and centrifuge to remove the supernatant to obtain purified silica-coated gold nanorods (SiO 2 @Au NRs). Prepared SiO 2 @Au NRs has significant absorption bands belonging to nano-gold at 720nm and 515nm, and compared with Au NRs, the absorption band shows a significant blue shift. Prepared SiO 2 The absorption spectrum of @Au NRs is as Image 6 C sh...

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Abstract

The invention relates to a fusion protein SZ1 for targeted therapy of colorectal cancer caused by excessive activation of Rspo, and a coupled nano-drug thereof, and belongs to the field of biological medicine. The efficient fusion protein SZ1 can effectively inhibit all four Rspo subtypes and can comprehensively characterize theSZ1 to inhibit Rspo-mediated key cell signal channels for promoting the occurrence and development of colorectal cancer and the growth of corresponding tumors at the levels of cells, organoids, experimental animals and the like. Meanwhile, a nano-drug loading technology and a tumor cell targeting technology are combined to prepare a coupled nano-drug of the fusion protein SZ1 of different materials, and it is confirmed that the coupled nano-drug of the fusion protein SZ1 can more effectively inhibit the activity of all the four Rspo subtypes, so that the stability of the fusion protein SZ1 is further improved, the targeting property of the coupled nano-drug of the fusion protein SZ1 to tumors is enhanced, tumor tissues can be more effectively inhibited and killed in vivo, and a better treatment effect is achieved.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a fusion protein SZ1 for targeted treatment of colorectal cancer caused by excessive activation of Rspo and a nano-medicine coupled thereto. Background technique [0002] Targeted therapy as an emerging anti-tumor therapy has received extensive attention in recent years. This treatment method is different from traditional surgery, radiotherapy and chemotherapy. It can accurately and effectively kill tumor tissues with specific targets, while causing less damage to normal tissues. The theory and concept of targeted therapy originate from the continuous deepening of people's research on the etiology of malignant tumors. Malignant tumors, as major diseases that endanger human beings, are all generated from uncontrolled excessive cell proliferation caused by gene mutations. However, tumors of different organs, tumors of different types in the same organoid, and even tumors of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C07K1/14C12N9/00C12N15/62C12N15/867A61K38/53A61K38/17A61K33/242A61K47/52A61K47/64A61K47/69A61P35/00
CPCC12N9/93C07K14/723C12N15/86C12Y603/02019A61K38/53A61K38/1796A61K33/242A61K47/64A61K47/52A61K47/6923A61P35/00C07K2319/02C07K2319/30C07K2319/20C07K2319/00C12N2740/15043C12N2800/107A61K2300/00
Inventor 盛韧郑少钦刘思宇张曦
Owner NORTHEASTERN UNIV
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