阅读说明：本技术 抗Claudin18.2抗体及其应用 (anti-Claudin18.2 antibody and application thereof ) 是由 杨阳 葛虎 陶维康 于 2020-03-31 设计创作，主要内容包括：本公开涉及抗Claudin18.2抗体及其应用。具体地,本发明涉及一种抗Claudin18.2抗体；包含所述抗Claudin18.2抗体CDR的鼠源抗体、嵌合抗体、人源化抗体及其抗原结合片段,以及其作为药物的用途。特别地,本公开涉及一种抗Claudin18.2抗体在制备用于治疗Claudin18.2阳性疾病或病症的药物中的用途。(The present disclosure relates to anti-claudin18.2 antibodies and uses thereof. In particular, the invention relates to an anti-claudin18.2 antibody; murine antibodies, chimeric antibodies, humanized antibodies and antigen binding fragments thereof comprising the anti-claudin 18.2 antibody CDRs and their use as medicaments. In particular, the disclosure relates to the use of an anti-claudin 18.2 antibody in the manufacture of a medicament for the treatment of a claudin18.2 positive disease or disorder.)

An anti-claudin18.2 antibody comprising a heavy chain variable region and a light chain variable region, wherein:

i) the heavy chain variable region comprises a heavy chain variable region and a heavy chain variable region as set forth in SEQ ID NO:3, and a light chain variable region comprising HCDR1, HCDR2, and HCDR3 having the same sequence as the heavy chain variable region set forth in SEQ ID NO:4, LCDR1, LCDR2 and LCDR3 having the same sequence as the light chain variable region; or

ii) the heavy chain variable region comprises a sequence identical to the sequence set forth in SEQ ID NO:5, and a light chain variable region comprising HCDR1, HCDR2, and HCDR3 having the same sequence as the heavy chain variable region set forth in SEQ ID NO:6, LCDR1, LCDR2 and LCDR 3.

The anti-claudin18.2 antibody of claim 1, comprising a heavy chain variable region and a light chain variable region, wherein:

iii) the heavy chain variable region comprises the amino acid sequences as set forth in SEQ ID NOs: 9. SEQ ID NO: 10 and SEQ ID NO: 11 HCDR1, HCDR2, and HCDR3,

the light chain variable region comprises the amino acid sequences shown as SEQ ID NO: 12. SEQ ID NO: 13 and SEQ ID NO: LCDR1, LCDR2 and LCDR3 shown at 14; or

iv) the heavy chain variable region comprises the amino acid sequences as set forth in SEQ ID NOs: 15. SEQ ID NO: 16 and SEQ ID NO: the HCDR1, HCDR2 and HCDR3 shown at 17,

the light chain variable region comprises the amino acid sequences shown as SEQ ID NO: 18. SEQ ID NO: 19 and SEQ ID NO: LCDR1, LCDR2 and LCDR3 shown at 20.

The anti-claudin18.2 antibody of claim 1 or 2, which is a murine, chimeric or humanized antibody.

The anti-claudin18.2 antibody of any of claims 1 to 3, comprising a heavy chain variable region and a light chain variable region, wherein:

v) the heavy chain variable region having at least 90% identity in its amino acid sequence to SEQ ID NO 3 or 24, and

the light chain variable region having at least 90% identity in its amino acid sequence to SEQ ID NO 4 or 21; or

vi) the heavy chain variable region having an amino acid sequence at least 90% identical to SEQ ID NO 5 or 31, and

the light chain variable region having at least 90% identity in its amino acid sequence to SEQ ID NO 6 or 28;

preferably:

1) the heavy chain variable region has an amino acid sequence shown as SEQ ID NO. 3 or at least 90% identity with the heavy chain variable region, and the light chain variable region has an amino acid sequence shown as SEQ ID NO. 4 or at least 90% identity with the light chain variable region;

2) the heavy chain variable region has an amino acid sequence shown as SEQ ID NO. 24 or at least 90% identity with the heavy chain variable region, and the light chain variable region has an amino acid sequence shown as SEQ ID NO. 21 or at least 90% identity with the light chain variable region;

3) the heavy chain variable region has an amino acid sequence shown as SEQ ID NO. 5 or at least 90% identity with the heavy chain variable region, and the light chain variable region has an amino acid sequence shown as SEQ ID NO. 6 or at least 90% identity with the light chain variable region; or

4) The heavy chain variable region has an amino acid sequence shown as SEQ ID NO. 31 or at least 90% identity with the heavy chain variable region, and the light chain variable region has an amino acid sequence shown as SEQ ID NO. 28 or at least 90% identity with the light chain variable region.

The anti-Claudin18.2 antibody of claim 4, which is a humanized antibody comprising framework regions derived from a human antibody or framework region variants thereof having from 1 to 10 back mutations on the light chain framework region and/or the heavy chain framework region, respectively, of a human antibody;

preferably, the framework region variant comprises a mutation selected from a) or b) below:

a) the light chain variable region comprises one or more back mutations selected from 22S, 85I and 87H, and/or the heavy chain variable region comprises one or more back mutations selected from 48I, 82T and 69M; or

b) The light chain variable region comprises one or more back mutations selected from 4L and 22S, and/or the heavy chain variable region comprises one or more back mutations selected from 38K, 40R, 48I, 66K, 67A, 69L, 71L and 73K;

more preferably, the framework region variant comprises a mutation selected from a-1) or b-1) below:

a-1) a back mutation comprising 22S, 85I and 87H in the light chain variable region, and a back mutation comprising 48I and 82T in the heavy chain variable region; or

b-1) a back mutation of 4L in the light chain variable region;

in 82T of the heavy chain variable region, 82 is position 82A according to Kabat regulation.

The anti-claudin18.2 antibody of claim 3, comprising a heavy chain variable region and a light chain variable region as shown below:

vii) the heavy chain variable region sequence is shown in SEQ ID NO 3 and the light chain variable region sequence is shown in SEQ ID NO:4 is shown in the specification; or

viii) the heavy chain variable region sequence is set forth in SEQ ID NOs 24, 25, 26 or 27 and the light chain variable region sequence is set forth in SEQ ID NOs: 21. 22 or 23; or

ix) the heavy chain variable region sequence is shown as SEQ ID NO:5 and the light chain variable region sequence is shown as SEQ ID NO:6 is shown in the specification; or

x) the heavy chain variable region sequence is shown as SEQ ID NO 31, 32, 33 or 34 and the light chain variable region sequence is shown as SEQ ID NO: 28. 29 or 30;

preferably, the anti-claudin18.2 antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region and a light chain variable region as shown below:

xi) the heavy chain variable region sequence is shown as SEQ ID NO. 31 and the light chain variable region sequence is shown as SEQ ID NO. 29; or

xii) the heavy chain variable region sequence is shown as SEQ ID NO. 26 and the light chain variable region sequence is shown as SEQ ID NO. 23.

The anti-Claudin18.2 antibody of any one of claims 1 to 6, further comprising a heavy chain constant region and a light chain constant region;

preferably, the heavy chain constant region is selected from the group consisting of human IgG1, IgG2, IgG3, and IgG4 constant regions, and variants thereof,

preferably, the light chain constant region is selected from human antibody kappa and lambda chain constant regions and variants thereof;

more preferably, the anti-Claudin 18.2 antibody comprises a heavy chain constant region as set forth in SEQ ID NO. 7 and a light chain constant region as set forth in SEQ ID NO. 8;

most preferably, the anti-claudin18.2 antibody comprises: a heavy chain having at least 90% identity to SEQ ID NO 35 or 42 and a light chain having at least 90% identity to SEQ ID NO 36 or 39; or

A heavy chain having at least 90% identity to SEQ ID NO 37 or 49 and a light chain having at least 90% identity to SEQ ID NO 38 or 46.

The anti-claudin18.2 antibody according to any of claims 1 to 7, comprising:

c) 35 and the heavy chain of SEQ ID NO:36, a light chain;

d) 42, 43, 44 or 45 and the heavy chain of SEQ ID NO: 39. 40 or 41;

e) 37 and the heavy chain of SEQ ID NO: 38; or

f) 49, 50, 51 or 52 and the heavy chain of SEQ ID NO: 46. 47 or 48.

The anti-claudin18.2 antibody of any of claims 1 to 8, comprising:

the heavy chain shown as SEQ ID NO. 44 and the light chain shown as SEQ ID NO. 41; or

The heavy chain shown as SEQ ID NO. 49, and the light chain shown as SEQ ID NO. 47.

An isolated anti-claudin 18.2 antibody that competes for binding to human claudin18.2 with the anti-claudin 18.2 antibody of any one of claims 1 to 9.

A nucleic acid molecule encoding the anti-claudin18.2 antibody of any of claims 1 to 10.

A host cell comprising the nucleic acid molecule of claim 11.

An antibody drug conjugate formed by coupling the anti-claudin18.2 antibody according to any one of claims 1 to 10 with a cytotoxic drug.

A pharmaceutical composition comprising a therapeutically effective amount of an anti-claudin 18.2 antibody according to any one of claims 1 to 10, or a nucleic acid molecule according to claim 11, or an antibody drug conjugate according to claim 13, and one or more pharmaceutically acceptable carriers, diluents, buffers or excipients.

A method for immunodetection or assay of claudin18.2, the method comprising:

a step of contacting the anti-claudin18.2 antibody according to any of claims 1 to 10 with a sample to be tested.

A kit comprising an anti-claudin18.2 antibody according to any of claims 1 to 10.

A method of treating a disease associated with claudin18.2, said method comprising administering to a subject a therapeutically effective amount of an anti-claudin 18.2 antibody of any one of claims 1 to 10, or a nucleic acid molecule of claim 11, or an antibody drug conjugate of claim 13 or a pharmaceutical composition of claim 14, preferably said disease is a tumor;

more preferably, the disease is selected from: squamous cell carcinoma of head and neck, cancer of head and neck, brain cancer, glioma, glioblastoma multiforme, neuroblastoma, central nervous system cancer, neuroendocrine tumor, laryngeal cancer, nasopharyngeal cancer, esophageal cancer, thyroid cancer, malignant pleural mesothelioma, lung cancer, breast cancer, liver cancer, hepatoma, hepatocellular carcinoma, hepatobiliary cancer, pancreatic cancer, gastric cancer, gastrointestinal cancer, intestinal cancer, colon cancer, colorectal cancer, renal cancer, clear cell renal cell carcinoma, ovarian cancer, endometrial cancer, cervical cancer, bladder cancer, prostate cancer, testicular cancer, skin cancer, melanoma, leukemia, lymphoma, bone cancer, chondrosarcoma, myeloma, multiple myeloma, myelodysplastic syndrome, kunkenberg's tumor, myeloproliferative tumor, squamous cell carcinoma, ewing's sarcoma, systemic light chain amyloidosis, and merkel cell carcinoma;

more preferably, the lymphoma is selected from: hodgkin's lymphoma, non-hodgkin's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, primary mediastinal large B-cell lymphoma, mantle cell lymphoma, small lymphocytic lymphoma, large B-cell lymphoma rich in T-cells/histiocytes, and lymphoplasmacytic lymphoma;

the lung cancer is selected from: non-small cell lung cancer and small cell lung cancer;

the leukemia is selected from: chronic myeloid leukemia, acute myeloid leukemia, lymphocytic leukemia, lymphoblastic leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, and myeloid leukemia.