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Application of nucleic acid aptamer-drug conjugate PTK7-GEMs in preparation of drug for treating bladder cancer

A nucleic acid aptamer, bladder cancer technology, applied in the field of biomedicine

Pending Publication Date: 2022-02-11
THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no relevant research on whether PTK7 is highly expressed in bladder cancer clinical specimens and bladder cell lines, and whether aptamer-drug conjugates (APDC) have specific targets in bladder cancer tissues, thereby It is specific and can deliver drug molecules efficiently, but there is no related report

Method used

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  • Application of nucleic acid aptamer-drug conjugate PTK7-GEMs in preparation of drug for treating bladder cancer
  • Application of nucleic acid aptamer-drug conjugate PTK7-GEMs in preparation of drug for treating bladder cancer
  • Application of nucleic acid aptamer-drug conjugate PTK7-GEMs in preparation of drug for treating bladder cancer

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Embodiment Construction

[0017] experimental method:

[0018] 1. Cell culture

[0019] Human bladder cancer cells (BIU87, 5637, T24, EJ, RT4, J82, UM-UC-3, TCCSUP) and normal bladder urothelial cell line Svhuc were purchased from ATCC; 5637 and Svhuc were cultured with RPMI-1640 and F12K, respectively , the rest of the cells were cultured in DMEM medium containing 10% fetal bovine serum; cells were washed with Dulbecco's phosphate-buffered saline (D-PBS).

[0020] 2. Immunohistochemistry

[0021] Paraffin sections were cleared in xylene and rehydrated in ethanol solution. Then, the sections were heated at 120°C for 3 min in 0.01 citrate buffer (pH 6.0) to retrieve the antigen. Soak in 3% hydrogen peroxide solution for 20 minutes to block endogenous peroxidase activity, and seal the tissue with universal blocking serum for 20 minutes. Tissue sections were incubated with anti-PTK7 antibody (Abcam; 1:300 dilution) for 2 h at room temperature, and then incubated with biotin-labeled secondary antibody ...

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Abstract

The invention discloses an application of a nucleic acid aptamer-drug conjugate PTK7-GEMs in the preparation of a drug for treating bladder cancer. Experimental results show that the PTK7-GEMs are in targeted combination with PTK7 protein highly expressed on the surface of a tumor cell membrane through spatial conformation, and release gemcitabine under the action of intracellular phosphatase in a macropinocytosis mode, so that a cytotoxic effect is played, and a remarkable anti-cancer effect is shown in three animal tumor-bearing models. The invention relates to the first research for verifying PTK7 high expression in bladder cancer tissue specimens and various bladder cancer cells; similarly, the nucleic acid aptamer-drug conjugate of gemcitabine (PTK7-GEMs) is a first nucleic acid aptamer-drug conjugate of gemcitabine, and gemcitabine is accurately transferred to a tumor site in a targeted manner through high affinity and specificity of a target of PTK7 protein, so that the anti-cancer effect is accurately performed.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of nucleic acid aptamer-drug conjugate PTK7-GEMs in the preparation of drugs for treating bladder cancer. Background technique [0002] Bladder cancer is the most common malignant tumor of the urinary system. In the world, the incidence of bladder cancer ranks eleventh among malignant tumors. Its formation process is complex, and many genes are involved. It can be divided into non-muscle invasive bladder cancer NMIBC, muscle invasive bladder cancer (MIBC) and metastatic bladder cancer, of which 20-30% of new cases are MIBC. The treatment of MIBC is based on radical cystectomy and pelvic lymph node dissection. The postoperative recurrence rate is as high as 30-45%, and the 5-year survival rate is 45-66%. Micrometastasis is one of the main reasons for the failure of local treatment of MIBC; 50% of newly diagnosed MIBC patients have developed potentia...

Claims

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Application Information

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IPC IPC(8): A61K47/54A61K31/7068A61P35/00A61P13/10A61P35/04
CPCA61K47/549A61K31/7068A61P35/00A61P13/10A61P35/04Y02P20/55
Inventor 刘建业向伟伍沛刘彪刘镓浩胡兴
Owner THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV
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