| Residential College | false |
| Patent Number | US/63684204 |
| Status | 申請中 Pending |
| ENGINEERED CD9 ANTIBODIES AND THEIR USE | |
| Year Issued | 2024 |
| 2024-08 | |
| Application Number | US/63684204 |
ZHAO QI ; Fu Shengyu
| |
| Country | United States |
| Subtype | 发明专利 Invention |
| Abstract | This disclosure provides compositions of various modified antibody constructs including single-chain variable fragments (scFvs) and humanized monoclonal antibodies derived from two newly characterized anti-CD9 monoclonal antibodies that specifically bind to human CD9 protein with high affinity. Also provided are methods for treating CD9-related diseases using the CD9 antibody construct of this invention to target CD9-expressing cells, including those of CD9-positive cancers, such as blood cancers including leukemia, especially B-cell acute lymphoblastic leukemia (B-ALL), as well as solid tumors. [0005] As such, in a first aspect, this invention provides an anti-CD9 antibody construct that specifically binds CD9 and comprises antibody heavy chain and light chain complementarity determining regions (CDRs) having the amino acid sequences of (1) SEQ ID NOs:5-7 and 8-10; or (2) SEQ ID NOs:11-13 and 14-16, respectively. The anti-CD9 antibody construct of this invention may be a polypeptide in the entirety, or it may be a polypeptide with at least one other polypeptide or a non-polypeptide component conjugated to it. [0006] In some embodiments, the CD9 antibody construct of this invention comprises an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL), each comprising the amino acid sequence set forth in (1) SEQ ID NOs:1 and 2; or (2) SEQ ID NOs:3 and 4, respectively. In some embodiments, the CD9 antibody construct of this invention comprises a VH and a VL, each comprising the amino acid sequence set forth in (1) SEQ ID NOs:20 and 21; or (2) SEQ ID NOs:22 and 23, respectively. In some embodiments, the CD9 antibody construct is an anti-CD9 single chain antibody or single chain variable fragment (scFv), for example, it specifically binds CD9 and comprises an antibody heavy chain and light chain variable regions comprising the CDRs having the amino acid sequences set forth in SEQ ID NOs:5-7 and 8-10 or SEQ ID NOs:11-13 and 14-16, respectively. In some embodiments, the antibody construct of this invention is a chimeric antigen receptor (CAR) construct generated from the anti-CD9 monoclonal antibodies or scFv disclosed above and herein, comprises the amino acid sequences set forth in SEQ ID NOs:1 and 2 or SEQ ID NOs:3 and 4 (humanized heavy chain and light chain variable region amino acid sequences) or SEQ ID NOs:20 and 21 or SEQ ID NOs:22 and 23 (murine antibody VH and VL amino acid sequences). In some embodiments, the CD9 antibody construct of this invention is a full-length IgG, a bispecific T cell engager (BiTE), or an antibody drug conjugate (ADC). In some embodiments, the CD9 antibody construct is a full-length IgG antibody comprising a modified (N297A substitution) antibody heavy chain constant region. For example, the anti-CD9 IgG antibody comprises the amino acid sequences set forth in SEQ ID NOs:5-7 and 8-10 or SEQ ID NOs:11-13 and 14-16 as the antibody heavy chain and light chain CDRs, respectively, preferably the amino acid sequences set forth in SEQ ID NOs:1 and 2 or SEQ ID NOs:3 and 4 as the antibody heavy chain and light chain variable regions, respectively, along with the amino acid sequence set forth in SEQ ID NO:17 as the antibody heavy chain constant region and the amino acid sequence set forth in SEQ ID NO:18 as the antibody light chain constant region. [0007] In a second aspect, the present invention provides a nucleic acid comprising a polynucleotide sequence encoding the anti-CD9 antibody construct, a fusion polypeptide comprising the anti-CD9 antibody construct described above and herein, for example, an anti-CD9 scFv or anti-CD9 CAR construct. The nucleic acid may comprise an expression cassette comprising a promoter operably linked to the polynucleotide sequence encoding the anti-CD9 antibody construct or its fusion polypeptide. In some cases, the nucleic acid is in the form of a vector, such as an expression vector including a plasmid or a viral vector. [0008] In some embodiments, the nucleic acid is contained and present within a host cell, either in the form of a free vector or in the form of DNA sequence permanently integrated into the host cell genome. The presence of such nucleic acid ensures the expression of the anti-CD9 antibody construct of this invention, e.g., an anti-CD9 scFv or anti-CD9 CAR construct. In some embodiments, the host cell is a T cell or natural killer (NK) cell. In some embodiments, the host cell is a T cell expressing a CAR construct comprising the amino acid sequence set forth in SEQ ID NOs:1 and 2 or SEQ ID NOs:3 and 4. [0009] In a third aspect, the present invention provides a conjugate comprising the CD9 antibody construct of the present invention. The conjugate includes a polypeptide component, i.e., the anti-CD9 antibody fusion polypeptide of this invention, and one or more conjugation partners, which may or may not be a protein in nature. For example, the conjugation partner may be a solid support, a detectable moiety, or a therapeutic agent. In some embodiments, the conjugate includes an anti-CD9 antibody construct, such as an anti-CD9 scFv, and a therapeutic agent, such as an anti-cancer agent. In this connection, compositions are also provided, which comprise the CD9 antibody construct of this invention, its encoding nucleic acid, including in the form of an expression cassette or vector, a host cell containing the nucleic acid and/or expressing the anti-CD9 antibody construct, or the conjugate comprising the anti-CD9 antibody fusion polypeptide of this invention joined with a conjugation partner, which may be another polypeptide or of a non-protein chemical nature. [0010] In a fourth aspect, the present invention provides a method for killing CD9-expressing cells. The method includes a step of contacting CD9-expressing cells, e.g., CD9-positive cancer cells, with a composition comprising an effective amount of an anti-CD9 antibody construct, which may or may not be conjugated with a therapeutic agent, e.g., anti-cancer agent, or an adequate number of T cells expressing the CD9 antibody CAR construct or other type of host cells expressing the anti-CD9 humanized antibodies described above and herein, e.g., comprising the amino acid sequence of SEQ ID NOs:1 and 2 or SEQ ID NOs:3 and 4. [0011] In some embodiments, the CD9+ cells being targeted by the claimed methods are cancer cells characterized by their CD9 expression, for example, leukemia (e.g., B-cell acute lymphoblastic leukemia or B-ALL) cells. In some embodiments, the cancer cells are located in a patient’s body. In some embodiments, the method further include these steps, prior to the step of contacting CD9-expressing cells (e.g., CD9-positive cancer cells) with a composition comprising an adequate number of T cells expressing the CD9 antibody CAR construct, (i) isolating T cells from a patient; (ii) transducing the T cells with the nucleic acid encoding the anti-CD9 CAR construct; (iii) cultivating the T cells ex vivo to expanding T cells expressing the anti-CD9 CAR construct comprising, from its N-terminus, an anti-CD9 scFv, a CD8 or CD28 transmembrane domain, a CD28 or 4-1BB costimulatory domain, and a CD3ζ T cell activating domain, and (iv) administering the expanded T cells in sufficient number back into the patient’s body. Exemplary CAR constructs comprise the amino acid sequence set forth in SEQ ID NOs:1 and 2 or SEQ ID NOs:3 and 4. In some embodiments, for the purpose of treating CD9-positive cancer, a patient in need for the treatment is administered an effective amount of an anti-CD9 antibody construct (e.g., optionally conjugated with an anti-cancer therapeutic agent) or T cells expressing the anti-CD9 CAR construct described above and herein, and the administering step comprises injection, e.g., by subcutaneous, intravenous, intramuscular, intraperitoneal, or intratumoral injection. [0012] In a fifth aspect, the present invention provides a kit for treating a disease or condition characterized by CD9 expression, for example, CD9+ cancers. The kit often includes a first container containing a first composition comprising an effective amount of an anti-CD9 antibody construct (e.g., optionally conjugated with a therapeutic agent) or an adequate number of host cells expressing an anti-CD9 antibody construct (such as T cells expressing a CAR comprising, for example, from its N-terminus, an anti-CD9 scFv, a CD8 or CD28 transmembrane domain, a CD28 or 4-1BB costimulatory domain, and a CD3ζ T cell activating domain), and a second container containing a second composition comprising an effective amount of another therapeutic agent known for its efficacy for treating the disease or condition, such as a chemotherapeutic or immunotherapeutic agent. In some embodiments, the CD9+ cancer is leukemia such as B-cell acute lymphoblastic leukemia (B-ALL). In some embodiments, the first or second composition is formulated for injection, e.g., for subcutaneous, intravenous, intramuscular, intraperitoneal, or intratumoral injection. In some cases, the kit may further comprises an instruction manual providing instructions for user to properly use the kit. |
| Document Type | Patent |
| Collection | DEPARTMENT OF BIOMEDICAL SCIENCES |
| Affiliation | University of Macau |
| Recommended Citation GB/T 7714 | ZHAO QI,Fu Shengyu. ENGINEERED CD9 ANTIBODIES AND THEIR USE. US/63684204[P]. 2024-08-01. |
| APA | ZHAO QI., & Fu Shengyu ENGINEERED CD9 ANTIBODIES AND THEIR USE. |
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