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Polypeptide molecular probe for targeting CD19 antibody and application of polypeptide molecular probe

A molecular probe and antibody technology, applied in the field of biomedicine, can solve the problems of in vivo transport process, tumor targeting ability, unknown proliferation and persistence in vivo, and difficult to control in vivo activity.

Pending Publication Date: 2022-08-05
JIANGSU INST OF NUCLEAR MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although CAR-modified T cell therapy targeting CD19 has achieved great success, it still faces many challenges, such as in vivo transport process, tumor targeting ability, in vivo proliferation and persistence, etc. are unknown, and in vivo activity is difficult to control

Method used

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  • Polypeptide molecular probe for targeting CD19 antibody and application of polypeptide molecular probe
  • Polypeptide molecular probe for targeting CD19 antibody and application of polypeptide molecular probe
  • Polypeptide molecular probe for targeting CD19 antibody and application of polypeptide molecular probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Preparation of GCGLKNRSSE-FITC

[0035] 1. The GCGLKNRSSE peptide was synthesized by solid-phase peptide synthesis. 2. Polypeptide GCGLKNRSSE is prepared by N-terminal modification of ACP linking group and then reacted with FITC to obtain GCGLKNRSSE-FITC. Its chemical structure is as follows figure 1 shown. like figure 2 shown, its molecular weight is 1552.68.

Embodiment 2

[0036] Example 2: In vitro targeting ability of polypeptide molecular probes

[0037] Reference Example 1 for the preparation of FITC-modified polypeptide molecular probes SPSGKLMSPK-FITC, RWNVSDLGGLGCGLKNRSSE-FITC, KDRPEIWEGE-FITC, GCGLKNRSSEGPSSPSGKLM-FITC, RWNVSDLGGLGCGLKNRSSEGPSSPSGKLM-FITC. Take the CART cells expressing antiCD19 scFv (provided by Shanghai Ucardi Biomedical Technology Co., Ltd.), count 10^6 cells in a 6-well plate with a cell counter, and add cells containing 50 micrograms of different polypeptide molecular probes. Medium solution 1mL. Co-incubate for 1 hour. The cells were aspirated and placed in a centrifuge tube and centrifuged at 1000 rpm for 5 minutes, the supernatant was discarded, fresh medium was added to resuspend, and this centrifugation and resuspension operation was repeated once. This cell suspension was analyzed with a BD flow cytometer. like image 3As shown, the positive rates of SEQ No. 2, 3, 5, and 6 in the CART cells expressing anti...

Embodiment 3

[0038] Example 3: RWNVSDLGGLGCGLKNRSSE-FITC-NOTA- 68 Preparation of Ga

[0039] 1. The preparation method of RWNVSDLGGLGCGLKNRSSE-FITC is the same as that of GCGLKNRSSE-FITC;

[0040] 2. RWNVSDLGGLGCGLKNRSSE-FITC was reacted with NOTA-MAL for 6 hours, and purified by semi-preparative high performance liquid chromatography to obtain RWNVSDLGGLGCGLKNRSSE-FITC-NOTA;

[0041] 3. 68 Ga labeling: Dissolve RWNVSDLGGLGCGLKNRSSE-FITC-NOTA in sodium acetate buffer (0.5mL, pH 6.8), add 2mL 68 GaCl 3 0.05M hydrochloric acid solution, the mixture was heated and reacted at 100 degrees Celsius for 10 minutes, purified by C18 solid phase extraction cartridge, rinsed with 1 mL of 50% ethanol, passed through a sterile filter into the product bottle, and diluted with 4 mL of physiological saline to obtain RWNVSDLGGLGCGLKNRSSE-FITC-NOTA- 68 Ga. Its chemical structure can be found in Figure 4 .

[0042] Radiochemical purity was determined by radio-HPLC. like Figure 5 As shown, the radi...

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Abstract

The invention discloses a polypeptide molecular probe of a targeted CD19 antibody and application of the polypeptide molecular probe. The polypeptide molecular probe is one or more of compounds with the following chemical structures, or pharmaceutically acceptable salts of the compounds, namely P1-Rn, P2-Rn, P3-Rn, P4-Rn, P5-Rn and P6-Rn. The molecular probe of the targeted CD19 antibody developed by the invention can trace the in-vivo transport process, tumor targeting ability, in-vivo proliferation and persistence of a cell therapy, is beneficial to monitoring the effectiveness and safety of the cell therapy in the whole process, and has important clinical value in the aspects of curative effect evaluation and therapeutic schedule guidance.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and particularly relates to a polypeptide molecular probe targeting CD19 antibody and its application. Background technique [0002] Relapsed and refractory acute lymphoblastic leukemia (ALL) remains a difficult disease to treat, and despite advances in upfront treatment and improved survival in de novo ALL, little improvement in outcomes has been seen for more than 20 years. Adoptive transfer of chimeric antigen receptor (CAR)-expressing T cells has emerged as a powerful targeted immunotherapy, showing striking responses in highly refractory populations. In children and adults with relapsed and refractory ALL, complete remission rates as high as 90% have been reported using CAR-modified T cells targeting the B-cell-specific antigen CD19. Although CAR-modified T-cell therapy targeting CD19 has achieved great success, it still faces many challenges, such as the in vivo transport process, tumo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K51/08A61K51/04A61K49/04A61K49/14A61K49/10G01N21/64A61K103/00
CPCA61K51/08A61K51/0482A61K49/04A61K49/14A61K49/106G01N21/6428
Inventor 王辛宇杨敏王广基潘栋辉严骏杰徐宇平王立振赵富宽张波
Owner JIANGSU INST OF NUCLEAR MEDICINE
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