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CN-122012512-A - Oligonucleotide aptamer specifically binding EGFR protein and application thereof

CN122012512ACN 122012512 ACN122012512 ACN 122012512ACN-122012512-A

Abstract

The invention provides an oligonucleotide aptamer specifically binding to EGFR protein and application thereof, belonging to the technical field of biological medicine. The invention provides a group of oligonucleotide aptamer sequences for specifically recognizing EGFR protein intracellular segments, which comprises EG2 and EG3 oligonucleotide aptamer sequences and modified sequences thereof, wherein the oligonucleotide aptamer can specifically recognize the EGFR protein intracellular segments, especially combine with recombinant proteins consisting of 668-1210 amino acids of EGFR protein, but not combine with other unrelated proteins, has potential closed target effect, can be applied to the field of research and development of new drugs based on EGFR pathway activation principle, and can be applied to research and development of related drugs for targeting EGFR.

Inventors

  • BAI CHENJUN
  • GU YONGQING
  • ZHOU PINGKUN
  • CHEN SHI
  • ZHANG TINGHUI

Assignees

  • 中国人民解放军军事科学院军事医学研究院

Dates

Publication Date
20260512
Application Date
20260226

Claims (10)

  1. 1. An oligonucleotide aptamer that specifically binds to an EGFR protein, wherein the oligonucleotide aptamer comprises at least one of EG2, EG3, an allosteric of EG2, and an allosteric of EG 3; the nucleotide sequence of EG2 is shown as SEQ ID No.1, and the nucleotide sequence of EG3 is shown as SEQ ID No. 2.
  2. 2. The oligonucleotide aptamer of claim 1, wherein the allosteric for EG2 comprises EG2S, EG M and EG2SF; the nucleotide sequence of EG2S is shown as SEQ ID No.3, the nucleotide sequence of EG2M is shown as SEQ ID No.4, and the nucleotide sequence of EG2SF is shown as SEQ ID No. 5.
  3. 3. The oligonucleotide aptamer of claim 1, wherein the allosteric for EG3 comprises EG3S; the nucleotide sequence of EG3S is shown in SEQ ID NO. 6.
  4. 4. The oligonucleotide aptamer of any one of claims 1-3, further comprising a modification of the oligonucleotide aptamer.
  5. 5. The oligonucleotide aptamer of claim 4, wherein the modification comprises a biotin modification at the 5' end.
  6. 6. Use of the oligonucleotide aptamer of any one of claims 1-5 in the preparation of a medicament for treating a disease associated with targeted intervention EGFR.
  7. 7. The use of claim 6, wherein the effect of the agent comprises at least one of inhibiting EGFR signaling pathway activation, mediating targeted drug delivery and enhancing therapeutic efficacy in combination with immunotherapy.
  8. 8. Use of the oligonucleotide aptamer of any one of claims 1-5 in the preparation of an EGFR diagnostic reagent.
  9. 9. Use of the oligonucleotide aptamer of any one of claims 1-5 in the preparation of an EGFR-targeting kit.
  10. 10. The use according to claim 9, wherein the kit type comprises at least one of an aptamer microplate detection kit, immunoprecipitation detection kit, an immunofluorescence detection kit, an aptamer-to-EIS electrical impedance sensor detection kit or an aptamer-to-nanogold chromogenic detection kit, an anti-tumor drug or a tumor therapeutic drug.

Description

Oligonucleotide aptamer specifically binding EGFR protein and application thereof Technical Field The invention belongs to the technical field of biological medicine, and particularly relates to an oligonucleotide aptamer specifically binding to EGFR protein and application thereof. Background The Aptamer (Aptamer) is a single-stranded DNA/RNA molecule screened by an exponential enrichment ligand system evolution technology (SELEX), can be combined with a target (such as a protein, a cell and the like) in a high specificity manner, and has the advantages of accurate targeting, low immunogenicity, easiness in modification and the like in tumor treatment. The method has long-term application direction and technical prospect in tumor treatment, and comprises the steps of (1) taking an aptamer as a guide head, and coupling a chemotherapeutic drug/a nucleic acid drug to realize accurate enrichment of tumor parts. It has been reported that AS1411 aptamer (targeting nucleolin) coupled with doxorubicin (Dox) has 3 times improved therapeutic effect and 60% reduced cardiotoxicity in breast cancer model. (2) Directly inhibit tumor signaling pathways, blocking key receptor-ligand interactions. ARC-01 aptamer has been reported to block PD-1/PD-L1 binding (IC 50 =2 nM), activate T cell killing, and treat melanoma ORR by 40% in clinical phase I. (3) regulating and controlling Tumor Microenvironment (TME), and relieving immunosuppression. It has been reported that CTLA-4 aptamer selectively binds to Treg cell CTLA-4, reversing immune tolerance (5-fold improvement over antibody penetration). EGFR (epidermal growth factor receptor) is used as a core target of tumor treatment, and the drug development of EGFR is expanded from a primary small molecule inhibitor to a multi-technology race track of double antibody, ADC, cell treatment and the like, and breakthrough progress is made in overcoming drug resistance and expanding indications. However, due to the problem of EGFR target mutation of tumor cells, the drug off-target effect still exists. The modified aptamer has high structural variability and stable combination, and meanwhile, the combination characteristic of the aptamer in a space three-dimensional structure mode is expected to be a breakthrough of a new generation of EGFR anti-tumor drugs. The aptamer medicine developed aiming at EGFR becomes a new strategy for overcoming the traditional treatment drug resistance by virtue of high affinity, low immunogenicity, easy tumor penetration and the like. Disclosure of Invention The invention provides an oligonucleotide aptamer specifically combined with EGFR protein and application thereof, wherein the oligonucleotide aptamer can specifically identify an EGFR protein intracellular segment and influence activation of a plurality of signal paths taking EGFR as an initial site, and can be widely applied to research of tumor therapeutic drug targets. The invention provides an oligonucleotide aptamer specifically binding to EGFR protein, wherein the oligonucleotide aptamer comprises at least one of EG2, EG3, an allosteric body of EG2 and an allosteric body of EG 3; the nucleotide sequence of EG2 is shown as SEQ ID No.1, and the nucleotide sequence of EG3 is shown as SEQ ID No. 2. In one embodiment of the invention, the allosteric bulk of EG2 comprises EG2S, EG M and EG2SF; the nucleotide sequence of EG2S is shown as SEQ ID No.3, the nucleotide sequence of EG2M is shown as SEQ ID No.4, and the nucleotide sequence of EG2SF is shown as SEQ ID No. 5. In one embodiment of the invention, the allosteric bulk of EG3 comprises EG3S; the nucleotide sequence of EG3S is shown in SEQ ID NO. 6. In one embodiment of the invention, the method further comprises modifying the oligonucleotide aptamer. In one embodiment of the invention, the modification comprises biotin modification at the 5' end. The invention also provides application of the oligonucleotide aptamer in preparing a medicament for treating EGFR related diseases in targeted intervention. In one embodiment of the invention, the effect of the agent comprises at least one of inhibiting EGFR signaling pathway activation, mediating targeted drug delivery and enhancing therapeutic efficacy in combination with immunotherapy. The invention also provides application of the oligonucleotide aptamer in preparation of EGFR diagnostic reagents. The invention also provides application of the oligonucleotide aptamer in preparation of a kit for targeting EGFR. In one specific embodiment of the invention, the kit comprises at least one of an aptamer micro-pore plate detection kit, a Immunoprecipitation detection kit, an immunofluorescence detection kit, an aptamer-combined EIS (electronic impedance system) electrical impedance sensor detection kit or an aptamer-combined nanogold chromogenic detection kit, an anti-tumor drug or a tumor therapeutic drug. The oligonucleotide aptamer has the beneficial effects that a group of oligonucleotide aptamer sequen