Cxcr3 ligand

Patent metadata

Field	Value
Publication number	US-2022017585-A1
Application number	US-201917299629-A
Country	US
Kind code	A1
Filing date	Dec 4, 2019
Priority date	Dec 4, 2018
Publication date	Jan 20, 2022
Grant date	—

Abstract

The present disclosure relates to CXCR3 ligands having resistance to DPPIV and having CXCR3-expressing cell migration-inducing activity, and specifically to N-terminal amino acid modifications and N-terminal amino acid sequences that are important for resistance to DPPIV and CXCR3-expressing cell migration-inducing activity.

First claim

1 . A CXCR3 ligand having resistance to DPPIV and having an activity to cause migration of cells expressing CXCR3.

2 . The CXCR3 ligand according to claim 1 , wherein the CXCR3 ligand has a C-X-C motif.

3 . The CXCR3 ligand according to claim 1 or 2 , wherein the CXCR3 ligand has any of the following sequences (a1) to (a7) at the N-terminus:
(a1) V-X1-L (X1 is F, G, I, K, L, M, T, V, W, or Y);
(a2) X2-V-P (X2 is A, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y);
(a3) V-X3-P (X3 is A, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y);
(a4) P-L-S;
(a5) X4-F-P (X4 is A, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, or Y);
(a6) F-X5-M (X5 is A, D, E, G, H, I, M, N, Q, R, S, T, V, W, or Y); and
(a7) F-X6-P (X6 is A, D, E, F, G, H, L, M, N, P, Q, S, T, W, or Y).

4 . The CXCR3 ligand according to any one of claims 1 to 3 , wherein the CXCR3 ligand has any of the following sequences (b1) to (b7) at the N-terminus:
(b1) V-X1-L-S-R-T-V-R (X1 is F, G, I, K, L, M, T, V, W, or Y);
(b2) X2-V-P-L-S-R-T-V-R (X2 is A, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y);
(b3) V-X3-P-L-S-R-T-V-R (X3 is A, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y);
(b4) P-L-S-R-T-V-R;
(b5) X4-F-P-M-F-K-R-G-R (X4 is A, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, or Y);
(b6) F-X5-M-F-K-R-G-R (X5 is A, D, E, G, H, I, M, N, Q, R, S, T, V, W, or Y); and
(b7) F-X6-P-M-F-K-R-G-R (X6 is A, D, E, F, G, H, L, M, N, P, Q, S, T, W, or Y).

5 . A CXCR3 ligand having any of the following sequences (a1) to (a7) at the N-terminus:
(a1) V-X1-L (X1 is F, G, I, K, L, M, T, V, W, or Y); (a2) X2-V-P (X2 is A, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y); (a3) V-X3-P (X3 is A, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y); (a4) P-L-S; (a5) X4-F-P (X4 is A, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, or Y); (a6) F-X5-M (X5 is A, D, E, G, H, I, M, N, Q, R, S, T, V, W, or Y); and (a7) F-X6-P (X6 is A, D, E, F, G, H, L, M, N, P, Q, S, T, W, or Y).

6 . The CXCR3 ligand according to claim 5 , which has any of the following sequences (b1) to (b7) at the N-terminus:
(b1) V-X1-L-S-R-T-V-R (X1 is F, G, I, K, L, M, T, V, W, or Y); (b2) X2-V-P-L-S-R-T-V-R (X2 is A, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y); (b3) V-X3-P-L-S-R-T-V-R (X3 is A, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y); (b4) P-L-S-R-T-V-R; (b5) X4-F-P-M-F-K-R-G-R (X4 is A, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, or Y); (b6) F-X5-M-F-K-R-G-R (X5 is A, D, E, G, H, I, M, N, Q, R, S, T, V, W, or Y); and (b7) F-X6-P-M-F-K-R-G-R (X6 is A, D, E, F, G, H, L, M, N, P, Q, S, T, W, or Y).

7 . The CXCR3 ligand according to claim 4 or 6 , which has a C-X-C motif at the C-terminus of the sequences (b1) to (b7).

8 . The CXCR3 ligand according to any one of claims 2 to 4 and 7 , wherein the CXCR3 ligand further has any of the following (c1) to (c5) at the C-terminus of the C-X-C motif:
(c1) the sequence from the 12th amino acid to the 77th amino acid of SEQ ID NO: 60;
(c2) the sequence from the 12th amino acid to the 73rd amino acid of SEQ ID NO: 61;
(c3) the sequence from the 12th amino acid to the 103rd amino acid of SEQ ID NO: 62;
(c4) the sequence from the 12th amino acid to the 77th amino acid of SEQ ID NO: 1; and
(c5) the sequence from the 12 th amino acid to the 77 th amino acid of SEQ ID NO: 63.

9 . The CXCR3 ligand according to any one of claims 1 to 9 , wherein the sequence of the CXCR3 ligand is any of the following (d1) to (d7):
(d1) a sequence shown by any one of SEQ ID NOs: 2 to 57, 92 to 147, and 149 to 204;
(d2) a sequence showing 90% or more sequence identity to SEQ ID NO: 60;
(d3) a sequence showing 90% or more sequence identity to SEQ ID NO: 61;
(d4) a sequence showing 90% or more sequence identity to SEQ ID NO: 62;
(d5) a sequence showing 90% or more sequence identity to SEQ ID NO: 63;
(d6) a sequence showing 90% or more sequence identity to SEQ ID NO: 1; and
(d7) a sequence comprising 10 or less amino acid substitutions, insertions, or deletions to a sequence selected from SEQ ID NOs: 1 to 57, 60 to 63, 92 to 147, and 149 to 204.

10 . A fusion protein comprising the CXCR3 ligand according to any one of claims 1 to 9 .

11 . A pharmaceutical composition comprising the CXCR3 ligand according to any one of claims 1 to 9 or the fusion protein according to claim 10 .

12 . A method of conferring resistance to DPPIV on a parent CXCR3 ligand in which the 2 nd amino acid from the N-terminus is P or A, wherein the method comprises any of the following:
(1) substituting the 2 nd amino acid from the N-terminus of the parent CXCR3 ligand from P or A to F, G, I, K, L, M, T, V, W, or Y; (2) further adding A, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y to the N-terminus of the parent CXCR3 ligand; (3) inserting A, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y between the 1 st and 2 nd amino acids at the N-terminus of the parent CXCR3 ligand; and (4) deleting V at the N-terminus of the parent CXCR3 ligand.

13 . The method according to claim 12 , wherein the N-terminal sequence of the parent CXCR3 ligand is V-P-L-S-R-T-V-R (SEQ ID NO: 86) or V-A-L-S-R-T-V-R (SEQ ID NO: 87).

14 . A method of producing a CXCR3 ligand having resistance to DPPIV, wherein the method carries out any of the following modifications to a parent CXCR3 ligand in which the 2 nd amino acid from the N-terminus is P or A:
(1) substituting the 2 nd amino acid from the N-terminus of the parent CXCR3 ligand from P or A to F, G, I, K, L, M, T, V, W, or Y; (2) further adding A, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, or Y to the N-terminus of the parent CXCR3 ligand; (3) inserting A, F, G, H, I, K, L, M, P, Q, R, S, T, V, W, or Y between the 1 st and 2 nd amino acids at the N-terminus of the parent CXCR3 ligand; and (4) deleting the V at the N-terminus of the parent CXCR3 ligand.

15 . The method according to claim 14 , wherein the N-terminal sequence of the parent CXCR3 ligand is V-P-L-S-R-T-V-R (SEQ ID NO: 86) or V-A-L-S-R-T-V-R (SEQ ID NO: 87).

16 . A method of conferring resistance to DPPIV on a parent CXCR3 ligand in which the 2 nd amino acid from the N-terminus is P, wherein the method comprises any of the following:
(1) further adding A, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, or Y to the N-terminus of the parent CXCR3 ligand; (2) substituting the 2 nd amino acid from the N-terminus of the parent CXCR3 ligand from P to A, D, E, G, H, I, M, N, Q, R, S, T, V, W, or Y; and (3) inserting A, D, E, F, G, H, L, M, N, P, Q, S, T, W, or Y between the 1 st and 2 nd amino acids at the N-terminus of the parent CXCR3 ligand.

17 . The method according to claim 16 , wherein the N-terminal sequence of the parent CXCR3 ligand is F-P-M-F-K-R-G-R (SEQ ID NO: 91).

18 . A method of producing a CXCR3 ligand having resistance to DPPIV, wherein the method carries out any of the following modifications to a parent CXCR3 ligand in which the 2 nd amino acid from the N-terminus is P:
(1) further adding A, D, E, F, G, H, I, K, L, M, N, P, Q, S, T, V, W, or Y to the N-terminus of the parent CXCR3 ligand; (2) substituting the 2 nd amino acid from the N-terminus of the parent CXCR3 ligand from P to A, D, E, G, H, I, M, N, Q, R, S, T, V, W, or Y; and (3) inserting A, D, E, F, G, H, L, M, N, P, Q, S, T, W, or Y between the 1 st and 2 nd amino acids at the N-terminus of the parent CXCR3 ligand.

19 . The method according to claim 18 , wherein the N-terminal sequence of the parent CXCR3 ligand is F-P-M-F-K-R-G-R (SEQ

Assignees

• Chugai Pharmaceutical Co Ltd

Inventors

• Yamamoto Yohei
• Sakurai Mika
• Haraya Kenta

Classifications

• C07K14/522

  Alpha-chemokines, e.g. NAP-2, ENA-78, GRO-alpha/MGSA/NAP-3, GRO-beta/MIP-2alpha, GRO-gamma/MIP-2beta, IP-10, GCP-2, MIG, PBSF, PF-4, KC · CPC title

• C07K2319/30

  Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto · CPC title

• C12P21/02

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• C07K14/521 Primary

  Chemokines · CPC title