WO2022120943A1 - 抗cll1抗体及其应用 - Google Patents
抗cll1抗体及其应用 Download PDFInfo
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- WO2022120943A1 WO2022120943A1 PCT/CN2020/138286 CN2020138286W WO2022120943A1 WO 2022120943 A1 WO2022120943 A1 WO 2022120943A1 CN 2020138286 W CN2020138286 W CN 2020138286W WO 2022120943 A1 WO2022120943 A1 WO 2022120943A1
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- variable region
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- antibody
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Classifications
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Definitions
- the present application belongs to the technical field of biomedicine, and relates to anti-CLL1 antibodies and applications thereof.
- CLL1 C-type lectin-like molecule 1
- CLEC12A C-type lectin domain family 12 member A
- DCAL-2 C-type lectin domain family 12 member A
- CLL1 C-type lectin domain family 12 member A
- the identification codes of CLL1 gene in some mainstream databases are as follows: Q5QGZ9 (UniprotKB), 31713 (HGNC), 160364 (Entrez Gene), ENSG00000172322 (Ensembl) and (612088) OMIM.
- CLL1 is restricted to hematopoietic cells, including mainly myeloid-derived cells in peripheral blood and bone marrow, such as monocytes, dendritic cells, granulocytes, and most acute myeloid leukemia (AML) cells. It is worth noting that although CLL1 is abundantly expressed on myeloid cells in peripheral blood and bone marrow, it is not expressed on myeloid-derived cells in peripheral tissues, such as tissue macrophages and tissue dendritic cells. .
- CLL1 is expressed on AML stem cells (CD34+/CD38-) and a small subset of hematopoietic progenitor cells (CD34+/CD38+ or CD34+/CD33+), but not on normal hematopoietic stem cells (CD34+/CD38- or CD34+/CD33-) not express. Due to this special expression pattern, CLL1 is expected to be a potential diagnostic and therapeutic target for AML.
- the currently known human CLL1 gene includes 7 transcripts, 5 of which encode proteins.
- Human CLL1 protein is a membrane receptor.
- the extracellular segment of the classical structure has a C-type lectin domain, and the intracellular segment has an immune receptor.
- Tyrosine motif (ITIM) phosphorylated ITIM binds to SH2 domain-containing phosphatase and negatively regulates the function of granulocytes and monocytes.
- the application provides anti-CLL1 antibodies and uses thereof for the treatment of cancer and autoimmune diseases, alone and/or in combination with other drugs.
- the application provides an anti-CLL1 antibody comprising a heavy chain variable region and a light chain variable region; wherein
- the heavy chain variable region comprises the CDR3 shown in SEQ ID NO:3, SEQ ID NO:9 or SEQ ID NO:14;
- the light chain variable region includes the CDR3 shown in SEQ ID NO:6, SEQ ID NO:11 or SEQ ID NO:17.
- the heavy chain variable region further comprises CDR1 as set forth in SEQ ID NO:1, SEQ ID NO:7 or SEQ ID NO:12.
- the heavy chain variable region further comprises the CDR2 set forth in SEQ ID NO:2, SEQ ID NO:8 or SEQ ID NO:13.
- the light chain variable region further comprises CDR1 as set forth in SEQ ID NO:4, SEQ ID NO:10 or SEQ ID NO:15.
- the light chain variable region further comprises the CDR2 set forth in SEQ ID NO:5 or SEQ ID NO:16.
- CDR1-3 of the heavy chain variable region of the antibody and CDR1-3 of the light chain variable region jointly determine the specific recognition and binding ability of the antibody to the antigen, including SEQ ID NO: 1-6, SEQ ID NO: 5 and the CDRs of SEQ ID NO: 7-11 or SEQ ID NO: 12-17 have significant binding ability to CLL1 protein.
- the heavy chain variable region of the anti-CLL1 antibody 23D7 includes CDR1 shown in SEQ ID NO:1, CDR2 shown in SEQ ID NO:2, and CDR3 shown in SEQ ID NO:3;
- the light chain variable region of the anti-CLL1 antibody 23D7 includes CDR1 shown in SEQ ID NO:4, CDR2 shown in SEQ ID NO:5, and CDR3 shown in SEQ ID NO:6; wherein
- the anti-CLL1 antibody 23D7 comprising the heavy chain variable region CDRs of SEQ ID NOs: 1 to 3 and the light chain variable region CDRs of SEQ ID NOs: 4 to 6 has CLL1 protein binding activity.
- the heavy chain variable region of the anti-CLL1 antibody 23D7 comprises the amino acid sequence set forth in SEQ ID NO: 18, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 19; wherein
- the heavy chain variable region of the anti-CLL1 antibody 19C1 includes CDR1 shown in SEQ ID NO:7, CDR2 shown in SEQ ID NO:8, and CDR3 shown in SEQ ID NO:9;
- the light chain variable region of the anti-CLL1 antibody 19C1 includes CDR1 shown in SEQ ID NO:10, CDR2 shown in SEQ ID NO:5, and CDR3 shown in SEQ ID NO:11; wherein
- the anti-CLL1 antibody 19C1 comprising the heavy chain variable region CDRs of SEQ ID NOs: 7 to 9 and the light chain variable region CDRs of SEQ ID NOs: 5 and 10 to 11 has CLL1 protein binding activity.
- the heavy chain variable region of the anti-CLL1 antibody 19C1 comprises the amino acid sequence set forth in SEQ ID NO: 20, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 21; wherein
- the heavy chain variable region of the anti-CLL1 antibody 27H4 includes CDR1 shown in SEQ ID NO:12, CDR2 shown in SEQ ID NO:13, and CDR3 shown in SEQ ID NO:14;
- the light chain variable region of the anti-CLL1 antibody 27H4 includes CDR1 shown in SEQ ID NO: 15, CDR2 shown in SEQ ID NO: 16, and CDR3 shown in SEQ ID NO: 17; wherein
- the anti-CLL1 antibody 27H4 comprising the heavy chain variable region CDRs of SEQ ID NOs: 12 to 14 and the light chain variable region CDRs of SEQ ID NOs: 15 to 17 has CLL1 protein binding activity.
- the heavy chain variable region of the anti-CLL1 antibody 27H4 comprises the amino acid sequence set forth in SEQ ID NO: 22, and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 23; wherein
- the anti-CLL1 antibody 27H4 is humanized, and the framework region of 27H4 is optimized to obtain a humanized H27H4 antibody that has a stronger affinity with CLL1, and the heavy chain variable region of the anti-CLL1 antibody H27H4 includes SEQ ID NO:
- the amino acid sequence shown in 24, the light chain variable region includes the amino acid sequence shown in SEQ ID NO: 25, SEQ ID NO: 26 or SEQ ID NO: 27; wherein
- the anti-CLL1 antibody is a monomer, formed by a set of heavy and light chains, and the variable regions of the heavy and light chains are linked by interchain disulfide bonds.
- the anti-CLL1 antibody is a multimer, formed by multiple groups of heavy chains and light chains, and the variable regions of the heavy chain and the light chain are connected by interchain disulfide bonds, and different heavy chains The variable regions are linked by interchain disulfide bonds.
- the anti-CLL1 antibody further comprises a constant region.
- the anti-CLL1 antibody is modified with a glycosylation group.
- the present application provides a nucleic acid molecule comprising a DNA fragment encoding the anti-CLL1 antibody of the first aspect.
- the present application provides an expression vector comprising the nucleic acid molecule of the second aspect.
- the present application provides a recombinant cell expressing the anti-CLL1 antibody of the first aspect.
- the nucleic acid molecule of the second aspect is integrated into the genome of the recombinant cell.
- the recombinant cell comprises the expression vector of the third aspect.
- the application provides a preparation method of the anti-CLL1 antibody described in the first aspect, the preparation method comprising the following steps:
- nucleic acid encoding the anti-CLL1 antibody is linked into a plasmid, transferred into competent cells, and cultured and then picked out of monoclonal cells for screening;
- the present application provides a pharmaceutical composition comprising the anti-CLL1 antibody of the first aspect.
- the pharmaceutical composition further includes an antitumor drug.
- the pharmaceutical composition further comprises any one or a combination of at least two pharmaceutically acceptable carriers, diluents or excipients.
- the present application provides the anti-CLL1 antibody of the first aspect, the nucleic acid molecule of the second aspect, the expression vector of the third aspect, the recombinant cell of the fourth aspect, or the sixth aspect.
- the disease comprises acute myeloid leukemia.
- the present application provides a method of treating cancer, the method comprising administering to a patient an effective dose of the anti-CLL1 antibody of the first aspect.
- the method further comprises administering one or more anti-tumor drugs simultaneously, separately or sequentially with the anti-CLL1 antibody.
- the cancer comprises acute myeloid leukemia.
- the anti-CLL1 antibodies 23D7, 27H4 and 19C1 of the present application have significant binding ability to CLL1, and the affinities of ch23D7, ch27H4 and ch19C1 for binding to the antigen CLL1 are 2.19nM, 3.83nM and 10.9nM respectively, which are comparable to the control antibody 1075.7;
- the anti-CLL1 antibodies 23D7, 27H4 and 19C1 of the present application can bind to the CLL1 protein on the cell surface, and the binding force increases with the increase of the antibody concentration;
- the antibody of the present application and its humanized modified antibody have important application prospects in the treatment of CLL1 positive tumors.
- Fig. 1 is the ForteBIO evaluation result of chimeric antibodies ch23D7, ch27H4 and ch19C1;
- Figure 2 shows the binding of chimeric antibodies ch23D7, ch27H4 and ch19C1 to CLL1 antigen by flow cytometry
- Figure 3 is a flow cytometry detection of the binding of humanized antibody hz27H4 to CLL1 antigen
- Figure 4 shows the ability of the antibody to bind to CLL1 transient 293T cells by flow cytometry
- FIG. 5A shows the results of MPA detection of the binding ability of 23D7-scFv-hFc to CLL1 antigen
- Figure 5B shows the results of MPA detection of the binding ability of 27H4-scFv-hFc to CLL1 antigen
- Figure 5C shows the results of MPA detection of the binding ability of 19C1-scFv-hFc to CLL1 antigen
- FIG. 5D is the MPA detection result of the binding ability of Hz27H4-scFv-hFc and CLL1 antigen
- FIG. 6A is a graph of the binding kinetics of recombinant human CLL-1
- FIG. 6B is a graph of the binding kinetics of recombinant cynomolgus monkey CLL-1.
- mice 10 BALB/C healthy female mice aged 7-8 weeks were selected, and the immunogen CLL1-mFc (fusion protein of CLL1 extracellular segment and mouse IgG1 Fc segment was used, and the amino acid sequence of CLL1 extracellular segment was as SEQ ID NO: 28) for immunization; two weeks after the second immunization, blood was collected from the tail vein of mice to separate serum, and ELISA was used to detect the antibody titer; two mice whose antibody titers reached the fusion requirement were selected three days before fusion.
- CLL1-mFc fusion protein of CLL1 extracellular segment and mouse IgG1 Fc segment was used, and the amino acid sequence of CLL1 extracellular segment was as SEQ ID NO: 28
- Blood was collected from the shock-immunized mice by cutting the neck. After sterilizing with 75% alcohol for 10 minutes, the spleen was removed, and the connective tissue was removed to prepare a spleen cell suspension. 5min, discard the supernatant, add RPMI1640 to 30mL, and count the cells; transfer the myeloma cells with good growth status (the number of viable cells>95%) to a 50mL centrifuge tube, add RPMI1640 to 30mL, centrifuge at 1000rpm for 5min, discard the supernatant , add RPMI1640 to 30mL, and count the cells;
- the splenocytes and myeloma cells were mixed in a ratio of 4:1, centrifuged at 1000 rpm for 5 min, the supernatant was discarded, the precipitated cell mass was bounced into a paste and placed in a 37°C water bath, 1 mL of fusion agent was added within 1 min and stirred well. Place in a 37°C water bath for 45-60s, add RPMI1640 within 1min to stop the fusion of the fusion agent, centrifuge at 1000rpm for 5min, and discard the supernatant;
- Antigen coating After diluting the pure antigen human CLL1 ECD-His (human CLL1 extracellular region with His tag) at a concentration of 50 ng/mL with the coating solution, take 100 ⁇ L and add it to the polystyrene enzyme-linked detection plate. In each well, overnight at 4°C;
- Reading measure the OD value of each well with a single wavelength of 450 nm, and select multiple clones with high reading values according to the principle of high to low for the next functional screening.
- the clones were sequenced to obtain the amino acid sequences of clones 23D7, 19C1, and 27H4, wherein the heavy chain variable region of 23D7 was shown in SEQ ID NO: 18, the light chain variable region was shown in SEQ ID NO: 19, and the variable region of 19C1 was shown in SEQ ID NO: 19.
- the heavy chain variable region is shown in SEQ ID NO:20, the light chain variable region is shown in SEQ ID NO:21, the heavy chain variable region of 27H4 is shown in SEQ ID NO:22, and the light chain variable region is shown in SEQ ID NO:22. shown in SEQ ID NO:23.
- the human-mouse chimeric antibody expression plasmid was transiently transfected into 293F cells, and chimeric antibodies ch23D7, ch19C1 and ch27H4 with 23D7, 19C1 and 27H4 as parents were obtained by transient expression and affinity purification.
- the ForteBio affinity measurement method (P.Estep et al., High throughput solution-based measurement of antibody-antigen affinity and epitope binning. MAbs, 2013.5(2):270-278.) was used to measure the chimeric antibodies ch23D7, ch19C1, ch27H4 Affinity testing was performed and a control antibody (Anti-CLL1-Ref) selected the VL and VL chains (SEQ ID NO: 29) of anti-human CLL-1 antibody 1075.7 (patent US8536310B2).
- the antibody was loaded onto an anti-human IgG capture (AHC) biosensor, the sensor was equilibrated offline in assay buffer for 30 min, and monitored online for 60 s to establish a baseline; the antibody-loaded sensor was combined with 100 nM of the antigen human CLL1 ECD- His was incubated for 5 min, then transferred to assay buffer, and the dissociation rate was measured after 5 min; kinetic analysis was performed using a 1:1 binding model.
- AHC anti-human IgG capture
- flow cytometry was used to detect the binding of antibodies ch23D7, ch27H4, and ch19C1 to CLL1 on HEK293 cells, and the steps were as follows:
- Control groups CLL1-RefAb (ch1075.7), Cell+secondary antibody and Blank were set.
- 27H4 was humanized.
- the gene sequence of the 27H4 antibody was compared with the human antibody Germline database to find sequences with high homology, while avoiding uncommon or small Germlines; after selecting a humanized template, rearranged
- the frequency of the amino acid occurrence of the antibody at a specific FR site, and CDR transplantation is performed to avoid the introduction of protein modification sites such as glycosylation and sites that are prone to chemical degradation; if the affinity of the sequence decreases after CDR transplantation, perform back mutation.
- the mutation principle is: Step-by-step single-point mutation is performed on the FR region; if there is a modification site or chemical degradation site in the CDR region, and the protein control is affected, the site is subjected to step-by-step single-point mutation; after each round of mutation, affinity (KD/Kon /Koff) detection; the finally obtained humanized sequence has the best performance in terms of affinity (within 3 times of the parental difference), stability, and protein quality (one-step PA>90%).
- one heavy chain variable region hz27H4H1 (SEQ ID NO:24) and three light chain variable regions hz27H4L1 (SEQ ID NO:25), hz27H4L2 (SEQ ID NO:26) and hz27H4L3 (SEQ ID NO:27).
- flow cytometry was used to detect the binding of antibodies hz27H4H1L1 and hz27H4H1L2 to CLL1 on HEK293 cells, and the steps were as follows:
- Control groups ch27H4, CLL1-RefAb (ch1075.7) and NC-huIgG1 were set.
- membrane proteome array (Membrane Proteome Array, MPA) was used to verify the non-target binding interaction of the antibody.
- MPA Membrane proteome array
- MPA is a platform for the analysis of specific antibodies and other ligands targeting human membrane proteins, which can be used to determine the specificity of antibody targets.
- Plasmids containing about 6000 membrane protein clones were transfected into HEK-293T cells (ATCC, CRL-3216) or QT6 cells (ATCC, CRL-1708), respectively, according to 18000 The density of cells/well was seeded into 384-well cell culture plates (Corning, 3764); after 36 hours of incubation, the test antibodies were added to the membrane proteome array matrix plates at predetermined concentrations, and the antibody scFv was directly detected by flow cytometry Binding to cells expressing about 6000 membrane proteins. All target proteins have native conformation and appropriate post-translational modifications.
- the mammalian-expressed single-chain antibody has a VL-(G4S) 3 -VH structure, and the C-terminal fusion expresses human hIgG1-Fc.
- the specific information is shown in Table 4.
- FCGR1A, FCGR2B, FCGR3B are IgG Fc receptors (IgG Fc receptors).
- CLL-1 antigens recombinant human CLL-1, Acro, product number: CLA-H5245, batch number: 3413a-9B8F1-SQ; recombinant Cynomolgus monkey CLL-1, Acro, Cat. No.: CLA-H5263, Lot No.: 3765-2079F1-SS
- scFv single chain antibody
- scFv VL-(G4S) 3 -VH structure
- C-terminal fusion Human hIgG1-Fc is expressed, and the sample information table is shown in Table 5.
- Antibody Diluent (Ligand): Dilute the antibody to 5 ⁇ g/mL with 1 ⁇ HBS-EP+running buffer;
- Recombinant human CLL-1 dilution solution (analyte 1): take recombinant human CLL-1 (250 ⁇ g/mL) and dilute it to 50nM with running buffer, 2-fold serial dilution to obtain 50nM, 25nM, 12.5nM, 6.25nM, 3.125 nM, 0nM recombinant human CLL-1 dilution;
- Recombinant cynomolgus monkey CLL-1 dilution (analyte 2): take recombinant cynomolgus monkey CLL-1 (250 ⁇ g/mL) and dilute it to 50nM with running buffer, and obtain 50nM, 25nM, 12.5nM, 6.25nM by 2-fold serial dilution , 3.125nM, 0nM recombinant cynomolgus monkey CLL-1 dilution;
- the protein A chip was used for detection. 5 ⁇ g/mL antibody diluent was passed through the experimental flow path (Fc2, Fc4) at a flow rate of 10 ⁇ L/min, and the capture volume was about 454RU for 20s. After that, the flow rate was adjusted to 30 ⁇ L/min.
- Concentrations of recombinant human CLL-1 dilution (0, 3.125nM, 6.25nM, 12.5nM, 25nM, 50nM) passed through the surface of the experimental flow path (Fc2, Fc4) and the reference flow path (Fc1, Fc3) at the same time, binding The time was 85s, the dissociation time was 70s, and finally glycine solution (Glycine, pH 1.5) was added for 60s to regenerate the chip and enter the next cycle.
- the protein A chip was used for detection. 5 ⁇ g/mL antibody diluent was passed through the experimental flow path (Fc2, Fc4) at a flow rate of 10 ⁇ L/min. After capturing for 20s, the capture volume was about 454RU, and then the flow rate was adjusted to 30 ⁇ L/min.
- Concentrations of recombinant cynomolgus monkey CLL-1 dilution (0, 3.125nM, 6.25nM, 12.5nM, 25nM, 50nM) passed through the surface of the experimental flow path (Fc2, Fc4) and the reference flow path (Fc1, Fc3) at the same time, The binding time was 85s, the dissociation time was 70s, and finally glycine solution (Glycine, pH 1.5) was added for 60s to regenerate the chip and enter the next cycle.
- the data analysis software Evaluation Software 3.1 was used to analyze the test results, and the sensing signal collected from the sample test flow path was double-deducted from the reference flow path and the sample blank, and the kinetic "1:1" model was used for fitting.
- the kinetic parameters (ka: association rate; kd: dissociation rate; kD: binding dissociation equilibrium constant) of each batch of samples with shTNF- ⁇ were obtained.
- the kinetic fitting results of the binding of 6 antibodies to recombinant human CLL-1 are shown in Table 6 and Figure 6A.
- the kinetic fitting results of binding to recombinant cynomolgus CLL-1 are shown in Figure 6B.
- the anti-CLL1 antibodies 23D7, 27H4, and 19C1 of the present application have significant binding ability to CLL1, and after humanization transformation, the affinity of the antibody and CLL1 is further improved, which is useful in the clinical diagnosis and/or treatment of tumors. It has important application prospects.
- the present application illustrates the detailed method of the present application through the above-mentioned embodiments, but the present application is not limited to the above-mentioned detailed method, which does not mean that the present application must rely on the above-mentioned detailed method for implementation.
- Those skilled in the art should understand that any improvement to the application, the equivalent replacement of each raw material of the product of the application, the addition of auxiliary components, the selection of specific methods, etc., all fall within the scope of protection and disclosure of the application.
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Abstract
Description
样品编号 | 浓度(nM) | Response | KD(M) | Kon(1/Ms) | Kdis(1/s) | RMax |
ch23D7 | 100 | 0.6536 | 2.19E-09 | 1.30E+05 | 2.84E-04 | 0.6693 |
ch27H4 | 100 | 0.6341 | 3.83E-09 | 2.65E+05 | 1.01E-03 | 0.6237 |
ch19C1 | 100 | 0.6194 | 1.09E-08 | 1.80E+05 | 1.97E-03 | 0.6574 |
ch1075.7 | 100 | 0.2361 | 4.14E-10 | 1.08E+05 | 4.48E-05 | 0.2384 |
样品编号 | 浓度(nM) | Response | KD(M) | Kon(1/Ms) | Kdis(1/s) |
hz27H4H1L1 | 60 | 0.6123 | 2.29E-09 | 3.77E+05 | 8.62E-04 |
hz27H4H1L2 | 60 | 0.7255 | 2.21E-09 | 4.41E+05 | 9.76E-04 |
hz27H4H1L3 | 60 | 0.7224 | 2.49E-09 | 4.38E+05 | 1.09E-03 |
ch27H4 | 60 | 0.5972 | 3.88E-09 | 3.20E+05 | 1.24E-03 |
Anti-CLL1 Ref Ab | 60 | 0.2189 | 6.76E-09 | 1.48E+05 | 9.97E-04 |
- | ch27H4 | hz27H4H1L1 | hz27H4H1L2 | CLL1-refAb |
EC50(μg/mL) | 0.3583 | 0.2618 | 0.3214 | 0.2446 |
抗体 | 基本信息 | 靶抗原 | Uniprot |
23D7-scFv-hFc | Anti-CLL1 scFv-hIgG1 | CLL1 | Q5QGZ9 |
27H4-scFv-hFc | Anti-CLL1 scFv-hIgG1 | CLL1 | Q5QGZ9 |
19C1-scFv-hFc | Anti-CLL1 scFv-hIgG1 | CLL1 | Q5QGZ9 |
Hz27H4-scFv-hFc | Anti-CLL1 scFv-hIgG1 | CLL1 | Q5QGZ9 |
样品 | ka(1/Ms) | kd(1/s) | KD(M) | Rmax(RU) | Chi 2(RU 2) |
27H4-scFv-hFc | 4.11E+06 | 1.26E-02 | 3.06E-09 | 249.8 | 0.839 |
19C1-scFv-hFc | 8.86E+05 | 5.09E-03 | 5.75E-09 | 273.8 | 0.11 |
23D7-scFv-hFc | 2.93E+05 | 3.22E-02 | 1.10E-07 | 272.1 | 0.232 |
Hz27H4-scFv-hFc | 4.12E+06 | 6.25E-03 | 1.52E-09 | 273.9 | 0.638 |
h27H4H1L1(全抗) | 4.91E+06 | 5.49E-03 | 1.12E-09 | 256.3 | 0.67 |
h27H4H1L2(全抗) | 5.04E+06 | 5.44E-03 | 1.08E-09 | 207.1 | 0.342 |
Claims (15)
- 抗CLL1抗体,其包括重链可变区和轻链可变区;其中所述重链可变区包括SEQ ID NO:3、SEQ ID NO:9或SEQ ID NO:14所示的CDR3;并且所述轻链可变区包括SEQ ID NO:6、SEQ ID NO:11或SEQ ID NO:17所示的CDR3。
- 根据权利要求1所述的抗CLL1抗体,其中,所述重链可变区还包括SEQ ID NO:1、SEQ ID NO:7或SEQ ID NO:12所示的CDR1。
- 根据权利要求1或2所述的抗CLL1抗体,其中,所述重链可变区还包括SEQ ID NO:2、SEQ ID NO:8或SEQ ID NO:13所示的CDR2。
- 根据权利要求1-3任一项所述的抗CLL1抗体,其中,所述轻链可变区还包括SEQ ID NO:4、SEQ ID NO:10或SEQ ID NO:15所示的CDR1。
- 根据权利要求1-4任一项所述的抗CLL1抗体,其中,所述轻链可变区还包括SEQ ID NO:5或SEQ ID NO:16所示的CDR2。
- 根据权利要求1-5任一项所述的抗CLL1抗体,其中,所述抗CLL1抗体的重链可变区包括SEQ ID NO:1所示的CDR1、SEQ ID NO:2所示的CDR2、SEQ ID NO:3所示的CDR3;并且所述抗CLL1抗体的轻链可变区包括SEQ ID NO:4所示的CDR1、SEQ ID NO:5所示的CDR2、SEQ ID NO:6所示的CDR3;或者所述抗CLL1抗体的重链可变区包括SEQ ID NO:7所示的CDR1、SEQ ID NO:8所示的CDR2、SEQ ID NO:9所示的CDR3;并且所述抗CLL1抗体的轻链可变区包括SEQ ID NO:10所示的CDR1、SEQ ID NO:5所示的CDR2、SEQ ID NO:11所示的CDR3;或者所述抗CLL1抗体的重链可变区包括SEQ ID NO:12所示的CDR1、SEQ ID NO:13所示的CDR2、SEQ ID NO:14所示的CDR3;并且所述抗CLL1抗体的轻链可变区包括SEQ ID NO:15所示的CDR1、SEQ ID NO:16所示的CDR2、SEQ ID NO:17所示的CDR3。
- 根据权利要求1-7任一项所述的抗CLL1抗体,其中,所述抗CLL1抗体的重链可变区包括SEQ ID NO:18所示的氨基酸序列,轻链可变区包括SEQ ID NO:19所示的氨基酸序列;或者所述抗CLL1抗体的重链可变区包括SEQ ID NO:20所示的氨基酸序列,轻链可变区包括SEQ ID NO:21所示的氨基酸序列;或者所述抗CLL1抗体的重链可变区包括SEQ ID NO:22所示的氨基酸序列,轻链可变区包括SEQ ID NO:23所示的氨基酸序列;或者所述抗CLL1抗体的重链可变区包括SEQ ID NO:24所示的氨基酸序列,轻链可变区包括SEQ ID NO:25、SEQ ID NO:26或SEQ ID NO:27所示的氨基酸序列。
- 根据权利要求1-7任一项所述的抗CLL1抗体,其中,所述抗CLL1抗体的重链可变区和轻链可变区之间通过链间二硫键连接;任选地,所述抗CLL1抗体的重链可变区之间通过链间二硫键连接;任选地,所述抗CLL1抗体还包括恒定区;任选地,所述抗CLL1抗体修饰有糖基化基团。
- 核酸分子,其包括编码权利要求1-8任一项所述的抗CLL1抗体的DNA片段。
- 表达载体,其包括权利要求9所述的核酸分子。
- 重组细胞,其表达权利要求1-8任一项所述的抗CLL1抗体。
- 根据权利要求11所述的重组细胞,其中,所述重组细胞的基因组中整合有权利要求6所述的核酸分子。
- 根据权利要求11所述的重组细胞,其中,所述重组细胞包括权利要求7所述的表达载体。
- 药物组合物,其包括权利要求1-8任一项所述的抗CLL1抗体;任选地,所述药物组合物还包括抗肿瘤药物;任选地,所述药物组合物还包括药学上可接受的载体、稀释剂或赋形剂中的任意一种或至少两种的组合。
- 权利要求1-8任一项所述的抗CLL1抗体、权利要求9所述的核酸分子、权利要求10所述的表达载体、权利要求11-13任一项所述的重组细胞或权利要求14所述的药物组合物在制备疾病检测试剂和/或疾病治疗药物中的应用;任选地,所述疾病包括急性髓系白血病。
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WO2024032247A1 (zh) * | 2022-08-09 | 2024-02-15 | 合源康华医药科技(北京)有限公司 | 一种cll1抗体及其应用 |
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WO2023201238A1 (en) * | 2022-04-11 | 2023-10-19 | Vor Biopharma Inc. | Binding agents and methods of use thereof |
CN115850476B (zh) * | 2022-08-09 | 2023-09-05 | 合源康华医药科技(北京)有限公司 | 一种cll1抗体及其应用 |
WO2024141063A1 (zh) * | 2022-12-30 | 2024-07-04 | 甘李药业股份有限公司 | 抗rsv病毒抗体、组合物、制剂及其应用 |
WO2024149225A1 (zh) * | 2023-01-10 | 2024-07-18 | 合源康华医药科技(北京)有限公司 | 一种人源化cll1抗体、嵌合抗原受体及其应用 |
CN116640211A (zh) * | 2023-02-28 | 2023-08-25 | 浙江康佰裕生物科技有限公司 | 特异性结合cll1蛋白的单域抗体及其应用 |
CN116462761B (zh) * | 2023-06-14 | 2023-09-08 | 浙江时迈药业有限公司 | 针对cll1的抗体及其用途 |
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