阅读说明：本技术 抗结缔组织生长因子抗体及其应用 (Anti-connective tissue growth factor antibody and application thereof ) 是由 付雅媛 马晓丽 葛虎 陶维康 于 2020-06-03 设计创作，主要内容包括：提供一种包含轻链和重链可变区的抗CTGF抗体及其作为药物的用途,该抗体可用于制备治疗CTGF相关疾病或病症的药物。(An anti-CTGF antibody comprising light and heavy chain variable regions useful for the preparation of a medicament for the treatment of a CTGF-associated disease or disorder and its use as a medicament are provided.)

An anti-CTGF antibody comprising a heavy chain variable region and a light chain variable region, wherein:

i) the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 having the same sequence as the heavy chain variable region shown in SEQ ID NO.6, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3 having the same sequence as the light chain variable region shown in SEQ ID NO. 7;

ii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 having the same sequence as the heavy chain variable region shown in SEQ ID NO:8 and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 having the same sequence as the light chain variable region shown in SEQ ID NO: 9;

ii-i) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 having the same sequence as the heavy chain variable region shown in SEQ ID NO:69 and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 having the same sequence as the light chain variable region shown in SEQ ID NO: 70; or

ii-ii) the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 having the same sequence as the heavy chain variable region shown in SEQ ID NO:85 and the light chain variable region comprises LCDR1, LCDR2, and LCDR3 having the same sequence as the light chain variable region shown in SEQ ID NO: 70.

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

iii) the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO 10, SEQ ID NO 11, and SEQ ID NO 12, respectively, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO 13, SEQ ID NO 14, and SEQ ID NO 15, respectively;

iv) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO 16, SEQ ID NO 17 and SEQ ID NO 18, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO 19, SEQ ID NO 20 and SEQ ID NO 21, respectively;

iv-i) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO 71, SEQ ID NO 72 and SEQ ID NO 73, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO 74, SEQ ID NO 75 and SEQ ID NO 76, respectively; or

iv-ii) the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO 102, SEQ ID NO 103 and SEQ ID NO 104, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO 74, SEQ ID NO 75 and SEQ ID NO 76, respectively.

The anti-CTGF antibody according to claim 1 or 2, wherein the anti-CTGF antibody is a murine antibody, a chimeric antibody, or a humanized antibody.

The anti-CTGF antibody according to any one 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 an amino acid sequence with at least 90% sequence identity to SEQ ID No.6, 27, 28 or 29 and the light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID No. 7, 22, 23, 24, 25 or 26;

(vi) the heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID No. 8, 33, 34, 35 or 36 and the light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID No.9, 30, 31 or 32; or

(vi-i) the heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO:69, 81, 82, 83, 84 or 85 and the light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO:70, 77, 78, 79 or 80.

The anti-CTGF antibody according to claim 2, wherein the anti-CTGF antibody is a humanized antibody comprising a framework region of a human antibody or a framework region variant thereof having up to 10 back mutations on the light chain framework region and/or the heavy chain framework region, respectively, of a human antibody;

preferably, the humanized antibody comprises a light chain variable region and a heavy chain variable region selected from (a), (b) or (b-i) below:

(a) a light chain variable region comprising:

LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO 13, SEQ ID NO 14 and SEQ ID NO 15, respectively, and one or more back mutations selected from 4L, 36F, 43S, 45K, 47W, 58V or 71Y, and

a heavy chain variable region comprising:

HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO 10, SEQ ID NO 11 and SEQ ID NO 12, respectively, and one or more back mutations selected from 28S, 30N, 49A, 75E, 76S, 93V, 94E or 104D;

(b) a light chain variable region comprising:

LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO 19, SEQ ID NO 20 and SEQ ID NO 21, respectively, and one or more back mutations selected from 36V, 44F, 46G or 49G, and

a heavy chain variable region comprising:

HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO 16, SEQ ID NO 17 and SEQ ID NO 18, respectively, and a heavy chain variable region back mutated with one or more amino acids selected from 44G, 49G, 27F, 48L, 67L, 71K, 78V or 80F;

(b-i) a light chain variable region comprising:

LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO 74, SEQ ID NO 75 and SEQ ID NO 76, respectively, and one or more back mutations selected from 45P, 46W, 48Y, 69S or 70Y, and

a heavy chain variable region comprising:

HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO 71, SEQ ID NO 72 and SEQ ID NO 73, respectively, and one or more back mutations selected from 27F, 38K, 48I, 67K, 68A, 70L or 72F: or

(b-ii) a light chain variable region comprising:

LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO 74, SEQ ID NO 75 and SEQ ID NO 76, respectively, and one or more back mutations selected from 45P, 46W, 48Y, 69S or 70Y, and

a heavy chain variable region comprising:

HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO 102, SEQ ID NO 103 and SEQ ID NO 104, respectively, and one or more back mutations selected from 27F, 38K, 48I, 67K, 68A, 70L or 72F.

The anti-CTGF antibody of claim 5 comprising a heavy chain variable region and a light chain variable region, wherein:

(vii) the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 6; and the variable region of the light chain, the amino acid sequence of which is shown as SEQ ID NO 7;

(viii) the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO 27, 28 or 29; and the variable region of the light chain, the amino acid sequence of which is shown as SEQ ID NO 22, 23, 24, 25 or 26;

(ix) the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 8; and the variable region of the light chain, the amino acid sequence of which is shown as SEQ ID NO 9;

(x) The amino acid sequence of the heavy chain variable region is shown as SEQ ID NO 33, 34, 35 or 36; and the variable region of the light chain, the amino acid sequence of which is shown as SEQ ID NO 30, 31 or 32;

(xi) The amino acid sequence of the heavy chain variable region is shown as SEQ ID NO: 69; and the variable region of the light chain, the amino acid sequence of which is shown as SEQ ID NO: 70; or

(xii) The amino acid sequence of the heavy chain variable region is shown as SEQ ID NO 81, 82, 83, 84 or 85; and the variable region of the light chain, the amino acid sequence of which is shown as SEQ ID NO 77, 78, 79 or 80;

preferably, it comprises a heavy chain variable region and a light chain variable region as shown below:

(xiii) The amino acid sequence of the heavy chain variable region is shown as SEQ ID NO: 27; and the variable region of the light chain, the amino acid sequence of which is shown in SEQ ID NO: 22;

(xiv) The amino acid sequence of the heavy chain variable region is shown as SEQ ID NO: 34; and the variable region of the light chain, the amino acid sequence of which is shown in SEQ ID NO: 30; or

(xv) The amino acid sequence of the heavy chain variable region is shown as SEQ ID NO: 85; and the amino acid sequence of the light chain variable region is shown as SEQ ID NO: 77.

The anti-CTGF antibody according to any one of claims 1 to 6, wherein the antibody further comprises antibody heavy chain constant regions and light chain constant regions; preferably, the heavy chain constant region is selected from the group consisting of constant regions of human IgG1, IgG2, IgG3, and IgG4, and variants thereof, and the light chain constant region is selected from the group consisting of constant regions of human kappa and lambda chains, and variants thereof; more preferably, the antibody comprises a heavy chain constant region as set forth in SEQ ID NO 37 or 38 and a light chain constant region as set forth in SEQ ID NO 39 or 40.

The anti-CTGF antibody according to any one of claims 1 to 7, comprising:

(c) a heavy chain having at least 85% sequence identity to SEQ ID NO 41, 43, 44, 45, 46, 47 or 48, and a light chain having at least 85% sequence identity to SEQ ID NO 42, 49, 50, 51, 52 or 53;

(d) a heavy chain having at least 85% sequence identity to SEQ ID NO 54, 56, 57, 58, 59, 60, 61, 62 or 63, and a light chain having at least 85% identity to SEQ ID NO 55, 64, 65 or 66;

(e) a heavy chain as set forth in SEQ ID NO 41, 43, 44, 45, 46, 47 or 48 and a light chain as set forth in SEQ ID NO 42, 49, 50, 51, 52 or 53;

(f) a heavy chain as set forth in SEQ ID NO 54, 56, 57, 58, 59, 60, 61, 62 or 63 and a light chain as set forth in SEQ ID NO 55, 64, 65 or 66;

(g) a heavy chain having at least 85% sequence identity to SEQ ID NO 86, 88, 89, 90, 91, 92, 93, 94, 95, 96 or 97 and a light chain having at least 85% sequence identity to SEQ ID NO 87, 98, 99, 100 or 101; or

(h) 86, 88, 89, 90, 91, 92, 93, 94, 95, 96 or 97 and 87, 98, 99, 100 or 101;

preferably, it comprises:

(j) the heavy chain shown as SEQ ID NO. 46 and the light chain shown as SEQ ID NO. 49;

(k) the heavy chain shown as SEQ ID NO. 61 and the light chain shown as SEQ ID NO. 64; or

(l) The heavy chain shown as SEQ ID NO. 97 and the light chain shown as SEQ ID NO. 98.

An isolated anti-CTGF antibody, wherein the isolated anti-CTGF antibody competes for binding to human CTGF with the anti-CTGF antibody of any one of claims 1 to 8.

A nucleic acid molecule encoding the anti-CTGF antibody of any one of claims 1 to 9.

A host cell comprising the nucleic acid molecule of claim 10.

A pharmaceutical composition comprising a therapeutically effective amount of an anti-CTGF antibody according to any one of claims 1 to 9, or a nucleic acid molecule according to claim 10, and one or more pharmaceutically acceptable carriers, diluents, buffers, or excipients.

A method for immunodetection or assay of CTGF, the method comprising the step of contacting the anti-CTGF antibody of any one of claims 1 to 9 with a subject or sample thereof.

A kit comprising an anti-CTGF antibody according to any one of claims 1 to 9.

A method for treating or preventing a CTGF-associated disease, the method comprising administering to a subject a therapeutically effective amount or a prophylactically effective amount of the anti-CTGF antibody of any one of claims 1 to 9, or the nucleic acid molecule of claim 10, or the pharmaceutical composition of claim 12, wherein the CTGF-associated disease is preferably a fibrous disease, hypertension, diabetes, myocardial infarction, arthritis, a CTGF-associated cell proliferative disease, atherosclerosis, glaucoma, or cancer;

preferably, the fibrotic disease is selected from: idiopathic pulmonary fibrosis, diabetic nephropathy, diabetic retinopathy osteoarthritis, scleroderma, chronic heart failure, cirrhosis of the liver, or renal fibrosis;

preferably, the cancer is selected from: acute lymphoblastic leukemia, cutaneous fibroids, breast cancer, angiolipoma, vascular leiomyoma, desmoplastic cancer, prostate cancer, ovarian cancer, colorectal cancer, pancreatic cancer, gastrointestinal cancer or liver cancer.