Polypeptides inhibiting CD40L

Patent metadata

Field	Value
Publication number	US-11248055-B2
Application number	US-201615778677-A
Country	US
Kind code	B2
Filing date	Nov 28, 2016
Priority date	Nov 27, 2015
Publication date	Feb 15, 2022
Grant date	Feb 15, 2022

Abstract

The present invention relates to immunoglobulins that specifically bind CD40L and more in particular to polypeptides, nucleic acids encoding such polypeptides; to methods for preparing such polypeptides; to compositions and in particular to pharmaceutical compositions that comprise such polypeptides, for prophylactic, therapeutic or diagnostic purposes. In particular, the immunoglobulins of the present invention inhibit the activity of CD40L and are safe.

First claim

The invention claimed is:

1. A polypeptide comprising at least one immunoglobulin single variable domain (ISVD) that binds CD40L, wherein said ISVD essentially consists of 4 framework regions (FR1 to FR4, respectively) and 3 complementarity determining regions (CDR1 to CDR3 respectively), in which:
(i) CDR1 is selected from the group consisting of SEQ ID NO: 33 and amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 33; CDR2 is selected from the group consisting of SEQ ID NO: 35 and amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 35; and CDR3 is selected from the group consisting of SEQ ID NO: 37 and amino acid sequences that have 1, 2, 3 or 4 amino acid difference(s) with SEQ ID NO: 37; or
(ii) CDR1 is selected from the group consisting of SEQ ID NO: 61 and amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 61; CDR2 is selected from the group consisting of SEQ ID NO: 63 and amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 63; and CDR3 is selected from the group consisting of SEQ ID NO: 65 and amino acid sequences that have 1, 2, 3 or 4 amino acid difference(s) with SEQ ID NO: 65; or
(iii) CDR1 is selected from the group consisting of SEQ ID NO: 40 and amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 40; CDR2 is selected from the group consisting of SEQ ID NO: 42 and amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 42; and CDR3 is selected from the group consisting of SEQ ID NO: 44 and amino acid sequences that have 1, 2, 3 or 4 amino acid difference(s) with SEQ ID NO: 44; or
(iv) CDR1 is selected from the group consisting of SEQ ID NO: 68 and amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 68; CDR2 is selected from the group consisting of SEQ ID NO: 70 and amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 70; and CDR3 is selected from the group consisting of SEQ ID NO: 72 and amino acid sequences that have 1, 2, 3 or 4 amino acid difference(s) with SEQ ID NO: 72.

2. The polypeptide according to claim 1 , in which CDR1 is chosen selected from the group consisting of
(a) SEQ ID NO: 61; and
(b) amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 61, wherein
at position 1 the G has been changed into E or R;
at position 2 the R has been changed into H or G;
at position 3 the T has been changed into I, A, S or P;
at position 4 the P has been changed into S;
at position 5 the L has been changed into P;
at position 6 the N has been changed into S, D or I;
at position 7 the Y has been changed into H;
at position 8 the H has been changed into N;
at position 9 the M has been changed into K, T or V; and/or
at position 10 the A has been changed into G, S or T;

and in which CDR2 is selected from the group consisting of
(a) SEQ ID NO: 63; and
(b) amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 63, wherein
at position 1 the A has been changed into G;
at position 2 the I has been changed into V;
at position 4 the S has been changed into N, R or G;
at position 6 the L has been changed into I;
at position 7 the G has been changed into S or D;
at position 8 the S has been changed into G, I or F; and/or
at position 9 the T has been changed into P or S;

and in which CDR3 is selected from the group consisting of
(a) SEQ ID NO: 65; and
(b) amino acid sequences that have 1, 2, 3 or 4 amino acid difference(s) with SEQ ID NO: 65, wherein
at position 1 the R has been changed into Q or L;
at position 2 the E has been changed into D or K;
at position 3 the T has been changed into S, M, A or K;
at position 4 the T has been changed into I, S, A or R;
at position 5 the H has been changed into Y or N;
at position 6 the Y has been changed into I, H or N;
at position 7 the S has been changed into T, G, N or I;
at position 8 the T has been changed into I or A;
at position 9 the S has been changed into N or R;
at position 10 the D has been changed into A;
at position 11 the R has been changed into S or G;
at position 13 the N has been changed into D, Y or S;
at position 14 the E has been changed into V, A, D or N;
at position 15 the M has been changed into I, V, K or T;
at position 16 the R has been changed into K, S, W, M, G or T;
at position 17 the H has been changed into N, L, Q, R or D;
at position 19 the D has been changed into N; and/or
at position 20 the Y has been changed into H, F or N.

3. The polypeptide according to claim 1 , in which:
CDR1 is SEQ ID NO: 33, CDR2 is SEQ ID NO: 35 and CDR3 is SEQ ID NO: 37; or
CDR1 is SEQ ID NO: 61, CDR2 is SEQ ID NO: 63 and CDR3 is SEQ ID NO: 65.

4. The polypeptide according to claim 1 , in which said ISVD is SEQ ID NO: 8 or SEQ ID NO: 6.

5. The polypeptide according to claim 1 , in which CDR1 is selected from the group consisting of
(a) SEQ ID NO: 40; and
(b) amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 40, wherein
at position 3 the T has been changed into S, N, A or I;
at position 4 the L has been changed into Q, S, M or G;
at position 8 the A has been changed into N or V;
at position 9 the I has been changed into L or V; and/or
at position 10 the G has been changed into A; and

in which CDR2 is selected from the group consisting of
(a) SEQ ID NO: 42; and
(b) amino acid sequences that have 1, 2 or 3 amino acid difference(s) with SEQ ID NO: 42, wherein
at position 2 the I has been changed into V;
at position 3 the S has been changed into G;
at position 5 the E has been changed into G;
at position 6 the G has been changed into S;
at position 7 the S has been changed into G, N, T or I;
at position 8 the T has been changed into A, P, I or S; and/or
at position 9 the S has been changed into I, R or G; and

in which CDR3 is selected from the group consisting of
(a) SEQ ID NO: 44; and
(b) amino acid sequences that have 1, 2, 3 or 4 amino acid difference(s) with SEQ ID NO: 44, wherein
at position 4 the R has been changed into S;
at position 7 the L has been changed into F, M or W;
at position 8 the G has been changed into D, A or S;
at position 9 the S has been changed into G, N or R;
at position 10 the S has been changed into G, N, T or R;
at position 12 the D has been changed into G, N, E or V;
at position 13 the T has been changed into N or A;
at position 14 the Q has been changed into H, K, L or R;
at position 15 the S has been changed into P or T;
at position 16 the H has been changed into N or Y;
at position 17 the Q has been changed into L, R or H;
at position 18 the Y has been changed into F;
at position 19 the D has been changed into G; and/or
at position 20 the Y has been changed into F or N.

6. The polypeptide according to claim 1 , in which:
CDR1 is SEQ ID NO: 40, CDR2 is SEQ ID NO: 42 and CDR3 is SEQ ID NO: 44; or
CDR1 is SEQ ID NO: 68, CDR2 is SEQ ID NO: 70, and CDR3 is SEQ ID NO: 72.

7. The polypeptide according to claim 1 , wherein said polypeptide binds to CD40L:
with a K D between 1E −07 M and 1E −13 M, between 1E −08 M and 1E −12 M, at most 1E −07 M, lower than 1E −08 M or 1E −09 M, or lower than 1E −10 M, 5E −11 M, 4E −11 M, 2E −11 M, 1.7E −11 M, 1E −11 , 5E −12 M, 4E −12 M, 3E −12 M, or 1E −12 M;
with an IC 50 between 1E −07 M and 1E −12 M, between 1E −08 M and 1E −11 M;
with an IC 50 of at most 1E −07 M, 1E −08 M, 1E −09 M, 5E −10 M, 4E −10 M, 3E −10 M, 2E −10 M, or 1E −10 M; and/or
with an off-rate of less than 5E −04 (s −1 ).

8. The polypeptide according to claim 1 , wherein said CD40

Assignees

• Ablynx Nv

Inventors

• Pattyn Els
• Van De Sompel Ariëlla
• Meerts Peter
• Buyse Marie-Ange
• Dewilde Maarten
• Beste Gerald
• Vlach Jaromir
• Hsu Jonathan

Classifications

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title

• C07K2317/569

  Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody® · CPC title

• C07K2317/567

  Framework region [FR] · CPC title

• A61K2039/505

  comprising antibodies · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title