DNA network hydrogel, preparation method and composition thereof

A hydrogel and composition technology, applied in the field of biopharmaceuticals, can solve the problems of high cost, autoimmune inflammation, etc., and achieve the effect of significant immune curative effect

Pending Publication Date: 2022-03-01
天津大学浙江研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Various macromolecular drugs currently developed, such as anti-PD-L1 antibodies, not only have the problem of high cost, but also may cause side effects such as autoimmune inflammation

Method used

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  • DNA network hydrogel, preparation method and composition thereof
  • DNA network hydrogel, preparation method and composition thereof
  • DNA network hydrogel, preparation method and composition thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A method for preparing a DNA network hydrogel encapsulating T lymphocytes, comprising the steps of:

[0036] DNA network hydrogel preparation

[0037] a. Design the antisense sequence (as shown in SEQIDNO.6) of the PD-1 DNA aptamer on the circular DNA template one, and design a DNA sequence complementary to a part of the circular DNA template on the circular DNA template two;

[0038] b. Proportionally, divide 5×10 -3 nmol of circular DNA template 1, 3U of phi29 DNA polymerase at a final concentration of 1×phi29 DNA polymerase buffer, a final concentration of 2×BSA, 0.05nmol of dNTPs, and a final concentration of 60mmol / L of NaCl in sterile water Make up to 100 μL; shake at 37°C for 4 hours at 350 rpm to obtain RCA product 1;

[0039] c. Proportionally, divide 5×10 -3nmol of circular DNA template II, 3 U of phi29 DNA polymerase at a final concentration of 1× phi29 DNA polymerase buffer, a final concentration of 2× BSA, 0.05 nmol of dNTPs, a final concentration of 60 ...

Embodiment 2

[0072] A method for preparing a DNA network hydrogel encapsulating T lymphocytes, comprising the steps of:

[0073] DNA network hydrogel preparation

[0074] a. Design the antisense sequence of the PD-1 DNA adapter (as shown in SEQ ID NO.6) to the circular DNA template one, and design a DNA sequence complementary to a part of the circular DNA template on the circular DNA template two ;

[0075] b. Proportionally, divide 1×10 -2 nmol of circular DNA template 1, 3.5U of phi 29 DNA polymerase, final concentration of 1×phi 29 DNA polymerase buffer, final concentration of 2×BSA, 0.06nmol of dNTPs, final concentration of 70mmol / L NaCl, with Make up to 100 μL with sterile water; shake at 36°C for 16 hours at 450 rpm to obtain RCA product 1;

[0076] c. Proportionally, divide 5×10 -3 ~1×10 -2 nmol of circular DNA template II, 3.5U of phi 29 DNA polymerase, final concentration of 1×phi 29 DNA polymerase buffer, final concentration of 2×BSA, 0.06nmol of dNTPs, final concentration o...

Embodiment 3

[0106] A method for preparing a DNA network hydrogel encapsulating T lymphocytes, comprising the steps of:

[0107] DNA network hydrogel preparation

[0108] a. Design the antisense sequence of the PD-1 DNA adapter (as shown in SEQ ID NO.6) to the circular DNA template one, and design a DNA sequence complementary to a part of the circular DNA template on the circular DNA template two ;

[0109] b. Proportionally, the 8×10 -3 nmol of circular DNA template 1, 4U of phi 29 DNA polymerase, final concentration of 1×phi 29 DNA polymerase buffer, final concentration of 2×BSA, 0.08nmol of dNTPs, final concentration of 80mmol / L NaCl, with no Make up to 100 μL with bacterial water; shake at 36°C for 8 hours at 400 rpm to obtain RCA product 1;

[0110] c. Proportionally, divide 5×10 -3 nmol of circular DNA template II, 3U of phi29 DNA polymerase at a final concentration of 1×phi 29 DNA polymerase buffer, a final concentration of 2×BSA, 0.05nmol of dNTPs, and a final concentration of ...

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Abstract

The invention discloses a DNA network hydrogel, a preparation method and a composition thereof, the network hydrogel comprises a network hydrogel carrier having a network structure formed by a DNA chain; and the HhaI (at) TGMS nano particles are loaded on the network hydrogel carrier. According to the invention, controllability of DNA network degradation and T cell release is endowed, so that blocking of immune checkpoints on cell surfaces and responsive release of tumor T lymphocytes are realized, and the nano-carrier is successfully applied to tumor immunotherapy and plays a remarkable immune curative effect in a melanoma mouse model.

Description

[0001] Prior filing date: 2021 / 11 / 4 [0002] Prior application number: 2021113016722 technical field [0003] The invention relates to biopharmaceutical technology, in particular to a DNA network hydrogel, a preparation method and a composition thereof. Background technique [0004] Immunotherapy is an emerging cancer treatment that aims to activate a patient's own immune system to attack tumor cells. Compared with traditional surgical therapy and chemotherapy, it has stronger systemic and lower systemic toxicity. In recent years, tumor immunotherapy has achieved major breakthroughs, but solid tumors have a tumor immunosuppressive microenvironment, which greatly reduces the effectiveness of existing immunotherapy techniques. For example, the programmed death ligand-1 (PD-L1) produced by tumor cells can bind to the PD-1 receptor on T cells and inhibit the activation of T cells, thereby reducing the efficacy of immunity. In recent years, researchers are constantly exploring...

Claims

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Application Information

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IPC IPC(8): C12N5/0783C12P19/34C12Q1/6844A61K35/17A61K47/26A61P35/00
CPCC12N5/0636C12P19/34C12Q1/6844A61K35/17A61K47/26A61P35/00C12N2509/00C12Q2525/205C12Q2525/307C12Q2521/101C12Q2531/125C12Q2521/501C12Q2525/131
Inventor 仰大勇姚池郭蕴华
Owner 天津大学浙江研究院
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