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CN-120943960-B - Specific monoclonal antibody targeting human CD180 and application thereof

CN120943960BCN 120943960 BCN120943960 BCN 120943960BCN-120943960-B

Abstract

The invention provides a specific monoclonal antibody targeting human CD180 and application thereof, and relates to the technical field of biological medicine. The targeted human CD180 antibody can realize specific binding with CD180 antigen and protein, can realize specific detection of CD180 in various tumor cell lines, and provides conditions for preparing a kit for detecting CD180 antigen. A CD180 chimeric antigen receptor (abbreviated as CD 180-CAR) was constructed based on the above antibodies, and CD180-CAR-T cells for tumor immune cell therapy were successfully constructed. The CD180-CAR-T cell provided by the invention has the capability of killing CD180 positive acute myeloid leukemia cells with high efficiency and specificity, can secrete high-level IFN gamma and IL-2 to resist CD180 positive cancer cells, can also remarkably eliminate tumor cells in mice, and has strong anti-tumor capability and life cycle prolonging capability. The specific monoclonal antibody targeting human CD180, the chimeric antigen receptor fusion protein targeting CD180 and the engineering immune cell targeting CD180 provide a new strategy for treating tumor of blood system.

Inventors

  • LI FENG
  • PENG YAJING
  • CHU YI
  • ZHANG YI

Assignees

  • 郑州大学第一附属医院

Dates

Publication Date
20260508
Application Date
20250804

Claims (7)

  1. 1. A specific monoclonal antibody targeting human CD180 is characterized in that the heavy chain variable region of the antibody is 28A7-VH, the 28A7-VH amino acid sequence is shown as SEQ ID NO. 1, and the light chain variable region of the antibody is 28A7-VL, and the 28A7-VL amino acid sequence is shown as SEQ ID NO. 4.
  2. 2. A specific monoclonal antibody targeting human CD180 is characterized in that the heavy chain of the antibody is 28A7-HEAVY CHAIN, the amino acid sequence of 28A7-HEAVY CHAIN is shown as SEQ ID NO. 7, and the light chain of the antibody is 28A7-LIGHT CHAIN, and the amino acid sequence of 28A7-LIGHT CHAIN is shown as SEQ ID NO. 10.
  3. 3. The specific monoclonal antibody targeting human CD180 according to claim 1, wherein the 28A7-scFv single chain variable fragment consists of 28A7-VH of the amino acid sequence shown in SEQ ID NO. 1 and 28A7-VL of the amino acid sequence shown in SEQ ID NO. 4, and the amino acid sequence of the 28A7-scFv is shown in SEQ ID NO. 13.
  4. 4. Use of a monoclonal antibody of any one of claims 1 to 3 that targets human CD180 for the preparation of a reagent for detecting CD180 antigen.
  5. 5. A chimeric antigen receptor that targets CD180, wherein the chimeric antigen receptor comprises from N-terminus to C-terminus: (i) 28A7-scFv targeting CD 180; (ii) A transmembrane domain; (iii) At least one co-stimulatory domain; (iv) An activation domain; the nucleotide sequence of the chimeric antigen receptor is shown as any one of SEQ ID NO 34 or SEQ ID NO 37.
  6. 6. A CD 180-targeting engineered immune cell expressing the CD 180-targeting chimeric antigen receptor of claim 5.
  7. 7. Use of a CD 180-targeting monoclonal antibody according to claim 1, a CD 180-targeting chimeric antigen receptor according to claim 5, a CD 180-targeting engineered immune cell according to claim 6 for the preparation of a medicament for the treatment of a CD 180-related cancer, wherein the cancer is acute myeloid leukemia.

Description

Specific monoclonal antibody targeting human CD180 and application thereof Technical Field The invention relates to the technical field of biological medicine, in particular to a specific monoclonal antibody targeting human CD180 and application thereof. Background Acute Myeloid Leukemia (AML) is the most common acute leukemia in adults and there is still no good treatment regimen. Standard induced chemotherapy can achieve complete remission in some AML patients, but eventually more than half AML patients will relapse and survival is poor. Chemotherapy resistance and short-term relapse remain major contributors to patient survival. Chimeric antigen receptor T cell (CAR-T) immunotherapy is a technology for realizing specific killing of tumor cells by expressing a chimeric antigen receptor molecule specifically recognizing and binding to a tumor antigen on a T cell membrane to induce T cell activation, and is one of the most promising tumor immunotherapy. In recent years, CAR-T cell therapy techniques have been widely used in clinical research as a promising therapeutic approach and have achieved good results. The approach to treating AML continues to evolve, however existing targets are poorly therapeutic, poorly released or can produce severe extra-tumor toxicity. There is therefore an urgent need to develop safer new targets for AML to achieve objective long-term relief. CD180 (also known as LY64 or RP 105) is a Toll-like receptor (TLR) family member, and is expressed mainly on the surface of B cells, dendritic cells, and various hematological tumor cells. Unlike other TLR members, the functional realization of CD180 is highly dependent on its heterodimeric complex with MD1 (Myeloid Differentiation Protein 1). Studies have shown that the interaction of CD180 with MD1 is critical for its stable expression and signaling on cell membranes. The data show that CD180 is highly expressed in various blood tumors such as acute myelogenous leukemia, diffuse large B cell lymphoma and the like, and is closely related to tumor cell proliferation, drug resistance and immune escape. At present, no treatment means aiming at CD180 exists, and development of high-specificity antibodies and immunotherapy based on the antibodies have great clinical value. CAR-T cell therapies have shown significant efficacy in hematological tumor treatment by genetically engineering T cells to express chimeric receptors targeting tumor antigens. However, existing CAR-T therapy targets (e.g., CD19, CD123, CD33, etc.) suffer from problems of antigen escape, high toxicity, or limited applicable population. The development of antibodies and CAR-T cells targeting CD180 can fill the gap, CD180 is used as a novel target, the indication range of CAR-T treatment can be expanded, and the kit is particularly suitable for patients with CD180 positive refractory blood tumor. Disclosure of Invention (One) solving the technical problems Aiming at the defects of the prior art, the invention provides a specific monoclonal antibody targeting human CD180 and application thereof in resisting tumor. (II) technical scheme In order to achieve the above purpose, the invention is realized by the following technical scheme: In a first aspect, the invention provides a specific monoclonal antibody (mAb) targeting human CD180, said antibody being abbreviated as antibody 28A7 or antibody 30A1 or antibody 38C8, respectively, said antibody being a mouse IgG1 subtype monoclonal antibody, the heavy chain variable region (VH) of said antibody comprising 28A7-VH or 30A1-VH or 38C8-VH, the light chain variable region (VL) of said antibody comprising 28A7-VL or 30A1-VL or 38C8-VL, the amino acid sequences of said 28A7-VH, 30A1-VH, 38C8-VH, 28A7-VL, 30A1-VL, 38C8-VL being shown in sequence in SEQ ID NOS 1-6. Specifically, the heavy chain of the antibody comprises 28A7-HEAVY CHAIN or 30A1-HEAVY CHAIN or 38C8-HEAVY CHAIN, the light chain of the antibody comprises 28A7-LIGHT CHAIN or 30A1-LIGHT CHAIN or 38C8-LIGHT CHAIN, and the amino acid sequence of 28A7-Heavy chain、30A1-Heavy chain、38C8-Heavy chain、28A7-Light chain、30A1-Light chain、38C8-Light chain is shown as SEQ ID NO 7-12 in sequence. Specifically, a 28A7-scFv single-chain variable fragment consists of 28A7-VH of the amino acid sequence shown in SEQ ID NO. 1 and 28A7-VL of the amino acid sequence shown in SEQ ID NO. 4, and the amino acid sequence of the 28A7-scFv is shown in SEQ ID NO. 13. Specifically, a 30A1-scFv single-chain variable fragment consists of 30A1-VH of an amino acid sequence shown in SEQ ID NO. 2 and 30A1-VL of an amino acid sequence shown in SEQ ID NO. 5, and the amino acid sequence of the 30A1-scFv is shown in SEQ ID NO. 14. Specifically, the 38C8-scFv single-chain variable fragment consists of 38C8-VH of the amino acid sequence shown in SEQ ID NO. 3 and 38C8-VL of the amino acid sequence shown in SEQ ID NO. 6, and the amino acid sequence of the 38C8-scFv is shown in SEQ ID NO. 15. Specifically, the complement