CN-121974910-A - Active small molecule with bio-orthogonal group and application thereof

Abstract

The invention discloses an active small molecule with a biological orthogonal group and application thereof, belonging to the technical field of biological medicine. The active small molecule is a small molecule hapten aiming at KRAS G12C mutant protein, and is formed by sequentially and covalently connecting tumor targeting molecules (AMG 510), PEG LINKER and bio-orthogonal groups (DBCO), wherein the chain length of PEG is 4, 10 or 16, the small molecule hapten can be specifically combined with the KRAS G12C protein inside and outside a cell, an artificial new antigen is formed by presenting an MHC-I molecule on the surface of a tumor cell after antigen processing, and the artificial new antigen is recruited and activated in situ by carrying out bio-orthogonal reaction with an Anti-human-CD3-N 3 antibody carrying an azide group, so that the accurate and efficient killing of the KRAS G12C mutant tumor cell is realized.

Inventors

• LIANG HONGWEN
• MA XUEDAN
• DANG YONGJUN
• WANG XIAOBO
• ZHANG FENGZHEN

Assignees

• 重庆医科大学

Dates

Publication Date

20260505

Application Date

20260123

Claims (2)

1. 1. An active small molecule with bio-orthogonal groups, characterized by the following formula: ; where n=4 or 10 or 16.
2. 2. The use of a small active molecule with bio-orthogonal groups according to claim 1 for the preparation of a medicament for the treatment of KRAS G12C mutation-related tumors.

Description

Active small molecule with bio-orthogonal group and application thereof Technical Field The invention relates to the technical field of biological medicine, in particular to an active small molecule with a biological orthogonal group and application thereof. Background The RAS gene is the first human proto-oncogene discovered, which encodes the RAS protein that plays a key role in cell signaling. Of these, KRAS G12C is the most common RAS mutation type, accounting for 44% of all RAS mutations. However, since RAS muteins are located inside cells, antibody drugs are difficult to target, and small molecule inhibitors are also prone to primary and secondary drug resistance, there is a need to develop novel therapeutic strategies against KRAS G12C. In recent years, neoantigen-mediated immunotherapy has provided a new approach to targeting intracellular muteins. The new antigen is derived from tumor specific mutation and is not expressed in normal tissues, so that the new antigen has high specificity and is an ideal accurate treatment target. By means of second generation sequencing and bioinformatics technology, researchers can utilize covalent binding of small molecule hapten and target protein to construct new antigen on the surface of tumor cells so as to activate immune response and overcome the drug resistance problem of the traditional inhibitor. However, a difficulty with neoantigen immunotherapy is how to accurately distinguish mutant peptides from wild-type peptides. Bio-orthogonal reactions offer the potential to achieve this goal as a highly efficient chemical and biological tool. Disclosure of Invention The invention aims to provide an active small molecule with a bio-orthogonal group and application thereof, and aims to solve the technical problems of drug resistance and difficulty in targeting intracellular proteins for immunotherapy existing in the existing KRAS G12C covalent inhibitor. In order to achieve the above purpose, the invention provides an active small molecule with bio-orthogonal groups, the structure of which is shown as the following formula: ; where n=4 or 10 or 16. On the other hand, the invention also provides application of the active small molecule with the bio-orthogonal group in preparing a medicament for treating KRAS G12C mutation related tumors. The invention designs a small molecule hapten which can be specifically combined with intracellular target protein KRAS G12C and carries the bio-orthogonal group, and the small molecule hapten can be used for marking target protein in cells, so that immune effector cells are recruited on the surfaces of tumor cells in situ by utilizing bio-orthogonal chemical reaction, thereby realizing accurate and efficient specific immune killing. The technical scheme of the invention is further described in detail through the drawings and the embodiments. Drawings In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the description of the embodiments of the present invention will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art. FIG. 1 is a coomassie brilliant blue staining chart of binding of small molecule haptens to KRAS G12C muteins inside and outside cells; FIG. 2 shows the results of western immunoblotting of the intracellular and extracellular binding of small molecule haptens to the KRAS G12C mutein; FIG. 3 shows the results of a cell membrane surface antigen presentation experiment; FIG. 4 shows the results of a T2 MHC stability test experiment; FIG. 5 shows the results of cell viability assay of CellTiter-Glo luminescence; FIG. 6 shows the result of crystal violet staining of small molecule hapten P4 in combination with Anti-human-CD3-N 3 after co-culture with MIAPaCa-2 cells; FIG. 7 is a schematic diagram of the structure and action of a small molecule hapten. Detailed Description The technical scheme of the invention is further described below through the attached drawings and the embodiments. In order to make the objects, technical solutions and advantages of the present application more clear, thorough and complete, the technical solutions of the present application will be clearly and completely described below through the accompanying drawings and examples. The following detailed description is of embodiments, and is intended to provide further details of the application. Unless defined otherwise, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The instrumentation and reagent materials used in the examples are all commercially available. The embodiment relates to small molecule haptens P4, P10 and P16 with different PEG chain lengths