IP Library › Granted Patent US 12,529,165
Granted Patent B2
US 12,529,165 · App. 17/867,434 · Granted Jan 20, 2026

Stabilized fibronectin based scaffold molecules

Inventor: Dasa Lipovsek (Pepperell, MA)
Assignee: Bristol-Myers Squibb Company
C40B40/10C07K2318/20
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Quick Facts
Patent No.
US 12,529,165
App. No.
17/867,434
Granted
Jan 20, 2026
Kind
B2
Abstract

Provided herein are proteins comprising a fibronectin based scaffold (FBS) domain, e.g., 10 Fn3 molecules, that bind specifically to a target, and wherein the FBS domain is linked at its C-terminus to a region consisting of PmXn, wherein P is proline, X is any amino acid and wherein n is 0 or an integer that is at least 1 and m is an integer that is at least 1, and wherein the PmXn moiety provides an enhanced property to the FBS domain, e.g., enhanced stability, relative to the protein that is not linked to the PmXn moiety.

Claims (16)

1 . An isolated multivalent fibronectin based scaffold (FBS) protein comprising two or more 10 Fn3 domains, wherein each of the 10 Fn3 domains comprises an amino acid sequence that is at least 70% identical to SEQ ID NO: 1, and wherein

(i) the C-terminal amino acid at the position corresponding to position 94 of SEQ ID NO: 1 of at least one 10 Fn3 domain is covalently linked through a peptide bond to a moiety consisting of the amino acid sequence PmXn, wherein P is proline, X is any amino acid, m is an integer that is at least 1 and n is 0 or an integer that is at least 1; and

(ii) the PmXn moiety provides an enhanced property to the multivalent FBS protein relative to the FBS protein that is not linked to the PmXn moiety.

2 . The isolated multivalent FBS protein of claim 1 , wherein the moiety covalently linked to the at least one 10 Fn3 domain consists of PmXn, wherein n is 1-10.

3 . The isolated multivalent FBS protein of claim 1 , wherein the moiety covalently linked to the at least one 10 Fn3 domain consists of PmXn, wherein n is 1-5.

4 . The isolated multivalent FBS protein of claim 1 , wherein the PmXn moiety covalently linked to the at least one 10 Fn3 domain consists of PI, PC, PID, PIE, PIDK (SEQ ID NO: 61), PIEK (SEQ ID NO: 63), PIDKP (SEQ ID NO: 65), PIEKP (SEQ ID NO: 67), PIDKPS (SEQ ID NO: 69), PIEKPS (SEQ ID NO: 71), PIDKPC (SEQ ID NO: 73), PIEKPC (SEQ ID NO: 75), PIDKPSQ (SEQ ID NO: 77), PIEKPSQ (SEQ ID NO: 79), PIDKPCQ (SEQ ID NO: 81), PIEKPCQ (SEQ ID NO: 83), PHHHHHH (SEQ ID NO: 87) or PCHHHHHH (SEQ ID NO: 86).

5 . The isolated FBS protein of claim 1 , wherein each of the least 10 Fn3 domains comprises the amino acid sequence LEVVAA(X)uLLISW(X)vYRITY(X)wFTV(X)xATISGL(X)yYTITVY A(X)ZISINYRT (SEQ ID NO: 16),

wherein (X)u, (X)v, (X)w, (X)x, (X)y and (X)z consist of the wild-type amino acid sequence (SEQ ID NO: 1) or comprise at least one amino acid difference with the corresponding wild-type sequence.

6 . The isolated multivalent FBS protein of claim 1 , wherein PmXn is P or PC.

7 . The isolated multivalent FBS protein of claim 1 , wherein at least one X of PmXn is a cysteine.

8 . The isolated multivalent FBS protein of claim 7 , wherein the cysteine is conjugated to a heterologous moiety.

9 . The isolated multivalent FBS protein of claim 8 , wherein the heterologous molecule is a detectable moiety.

10 . The isolated multivalent FBS protein of claim 8 , wherein the heterologous molecule is a drug moiety and the drug moiety and the multivalent FBS protein form an FBS-drug conjugate.

11 . The isolated FBS protein of claim 1 , wherein the enhanced property conferred by the PmXn moiety is enhanced stability.

12 . The isolated FBS protein of claim 11 , wherein enhanced stability is an increase in Tm of at least 1° C., 2° C., 3° C., 4° C., 5° C. or more.

13 . The isolated multivalent FBS protein of claim 6 , wherein PmXn is PC.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 22, 2022
From: LIPOVSEK, DASA
To: BRISTOL-MYERS SQUIBB COMPANY
Reel/Frame 060597/0922 →
Continuity (5)
Division 16560521 · Sep 4, 2019
Continuation 15127183
Provisional Application 62084270 · Nov 25, 2014
Provisional Application 61955975 · Mar 20, 2014
Related Publication 20230192812A1 · Jun 22, 2023
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