Granted Patent B2

US 12,517,136 · App. 16/827,001 · Granted Jan 6, 2026

Inventors: James R. Heath (Pasadena, CA); Heather Agnew (Culver City, CA); Blake Farrow (Pasadena, CA); David Bunck (Pasadena, CA); Jingxin Liang (Pasadena, CA); Arundhati Nag (Pasadena, CA); Samir Das (Pasadena, CA); Bert Tsunyin Lai (Culver City, CA); Suresh Mark Pitram (Culver City, CA)

Assignees: California Institute of Technology; Regeneron

G01N33/6869C07K7/06C07K7/64C07K14/54C07K2319/00G01N2333/54

Abstract

Claims (29)

1 . A method of producing a synthetic capture agent that binds to Interleukin 17F (IL-17F), the method comprising:

(a) selecting a first ligand that specifically binds to a first epitope on IL-17F, wherein the first epitope comprises the amino acid sequence FFQKPES (SEQ ID NO: 1), wherein the first ligand comprises the amino acid sequence selected from the group consisting of YFLLK (SEQ ID NO:5), FYKQH (SEQ ID NO:6), FYLTH (SEQ ID NO: 7), FYLQH (SEQ ID NO:8), RRATS (SEQ ID NO:9), and RRAQS (SEQ ID NO: 10), and wherein each of the first ligand is cyclic comprising 1,4-substituted-1,2,3-triazole residue (Tz4) or 1,5-substituted-1,2,3-triazole residue (Tz5);

(b) selecting a second ligand that specifically binds to a second epitope on IL-17F, wherein the second epitope comprises the amino acid sequence NENQRVS (SEQ ID NO:3), wherein the second ligand comprises the amino acid sequence selected from the group consisting of KYGEV (SEQ ID NO:11), LYGEV (SEQ ID NO: 12), VHKSG (SEQ ID NO:13), VHLSG (SEQ ID NO:14), QKHGP (SEQ ID NO: 15), TKHGP (SEQ ID NO:16), QLHGP (SEQ ID NO:17), TKHGP (SEQ ID NO: 18), YDLQR (SEQ ID NO:19), YDLTR (SEQ ID NO:20), YDKQR (SEQ ID NO: 21), YDKTR (SEQ ID NO:22), KKGWP (SEQ ID NO:23), KLGWP (SEQ ID NO: 24), LKGWP (SEQ ID NO:25), LLGWP (SEQ ID NO:26), RSYNL (SEQ ID NO: 27), or RSYNK (SEQ ID NO:28), and wherein each of the second ligand is cyclic comprising 1,4-substituted-1,2,3-triazole residue (Tz4) or 1,5-substituted-1,2,3-triazole residue (Tz5); and

(c) covalently linking the first ligand and the second ligand through a linker, thereby producing the synthetic capture agent that binds to IL-17F, wherein the linker has a length that allows both the first ligand and the second ligand to bind specifically to the first epitope and second epitope on IL-17F.

2 . The method of claim 1 , wherein the linker is selected by estimating the linker length by determining to which side of the first epitope the first ligand binds, to which side of the second epitope the second ligand binds, and measuring the distance between the bound sides of the first epitope and the second epitope on a folded structure of IL-17F.

3 . The method of claim 1 , wherein the linker is selected by screening a library of candidate linker molecules linking the first ligand and the second ligand for binding to IL-17F.

4 . The method of claim 1 , wherein the linker is selected by testing a candidate capture agent linked by a candidate linker for binding to IL-17F.

5 . The method of claim 1 , wherein the first epitope and the second epitope are from about 4.4 Å to about 26.4 Å, from about 8.8 Å to about 26.4 Å, from about 7 Å to about 15 Å, or about 15 Å distant from each other.

6 . The method of claim 1 , wherein the linker is 10-50% longer than the distance between the first epitope and the second epitope.

7 . The method of claim 1 , wherein the linker is 5-25% longer than the distance between the first epitope and the second epitope.

8 . The method of claim 1 , wherein the linker is 1-10% longer than the distance between the first epitope and the second epitope.

9 . The method of claim 1 , wherein the synthetic capture agent has a greater binding affinity for IL-17F than either of the first ligand or the second ligand.

10 . The method of claim 9 , wherein the synthetic capture agent has a binding affinity that is at least 50% of the binding affinity of a full cooperative binder.

11 . The method of claim 9 , wherein the synthetic capture agent has a binding affinity that is at least 75% of the binding affinity of a full cooperative binder.

12 . The method of claim 9 , wherein the synthetic capture agent has a binding affinity that is at least 90% of the binding affinity of a full cooperative binder.

13 . The method of claim 1 , wherein the first epitope and the second epitope are synthetic epitopes, wherein the synthetic epitopes each independently comprise at least a 20 amino acid sequence from a full length IL-17F, wherein at least one amino acid of each of the synthetic epitopes comprise an azide group or an acetylene group.

14 . The method of claim 13 , wherein the full length IL-17F is naturally occurring protein.

15 . The method of claim 13 , wherein the synthetic epitopes each independently comprise at least a 50 amino acid sequence from a full length IL-17F.

16 . The method of claim 13 , wherein the synthetic capture agent binds the synthetic epitopes and the full length IL-17F with a binding affinity that is at least 50% of the binding affinity of a full cooperative binder.

17 . The method of claim 13 , wherein the synthetic capture agent binds the synthetic epitopes and the full length IL-17F with a binding affinity that is at least 75% of the binding affinity of a full cooperative binder.

18 . The method of claim 13 , wherein the synthetic capture agent binds the synthetic epitopes and the full length IL-17F with a binding affinity that is at least 90% of the binding affinity of a full cooperative binder.

19 . The method of claim 1 , wherein the linker comprises one or more repeat units of ethylene glycol.

20 . The method of claim 19 , wherein the linker is PEG 1 , PEG 2 , PEG 3 , PEG 4 , or PEG 5 .

21 . The method of claim 1 , wherein the linker comprises a peptide.

22 . The method of claim 1 , wherein the linker comprises an amino acid.

23 . The method of claim 22 , wherein the amino acid is glycine.

24 . The method of claim 1 , wherein the linker comprises an alkylene moiety.

25 . The method of claim 1 , wherein selecting the first ligand is accomplished by screening a peptide library against a first synthetic epitope and identifying a first library peptide that is coupled to the first synthetic epitope during the screening, wherein the first synthetic epitope is a peptide comprising the sequence of the first epitope and a click handle, wherein the peptides of the first peptide library each comprise a click handle, wherein the click handle of the first synthetic epitope and the click handle of the first library peptides are (1) an azide click handle and an acetylene click handle, respectively, or (2) an acetylene click handle and an azide click handle, respectively.

26 . The method of claim 25 , wherein selecting the second ligand is accomplished by screening a second peptide library against a second synthetic epitope and identifying a second library peptide that is coupled to the second synthetic epitope during the screening, wherein the second synthetic epitope is a peptide comprising the sequence of the second epitope and a click handle, wherein the peptides of the second peptide library each comprise a click handle, wherein the click handle of the second synthetic epitope and the click handle of the second library peptides are (1) an azide click handle and an acetylene click handle, respectively, or (2) an acetylene click handle and an azide click handle, respectively.

Assignments (3)

Continuity (5)

References Cited (233)