Granted Patent
B2
US 12,534,511 · App. 17/527,238 · Granted Jan 27, 2026
Serum albumin-binding fibronectin type III domains
Inventors:
Tracy S. Mitchell (Andover, MA); Michael L. Gosselin (Boston, MA); Dasa Lipovsek (Pepperell, MA); Rex Parker (Titusville, NJ); Ray Camphausen (Wayland, MA); Jonathan H. Davis (Madison, WI); David Fabrizio (South Hamilton, MA)
Assignee:
Bristol-Myers Squibb Company
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Abstract
The present invention relates to polypeptides which include tenth fibronectin type III domains ( 10 Fn3) that binds to serum albumin, with south pole loop substitutions. The invention further relates to fusion molecules comprising a serum albumin-binding 10 Fn3 joined to a heterologous protein for use in diagnostic and therapeutic applications.
Claims (25)
1 . A fusion polypeptide comprising
(a) polypeptide comprising a fibronectin type III tenth domain ( 10 Fn3) wherein the 10 Fn3 domain comprises AB, BC, CD, DE, EF, and FG loops, wherein the amino acid sequence of the CD loop of the wild-type 10 Fn3 domain set forth in SEQ ID NO: 1 is replaced by an amino acid sequence selected from the group consisting of SEQ ID NOs: 101-125, and wherein the polypeptide binds to human serum albumin, and to one or more of rhesus serum albumin, cynomolgus serum albumin, mouse serum albumin, and rat serum albumin; and
(b) at least one heterologous protein, wherein the at least one heterologous protein is linked to the polypeptide with (i) a polypeptide linker having a protease site or (ii) a biocompatible polymer.
2 . The fusion polypeptide of claim 1 , wherein the 10 Fn3 domain of the polypeptide binds to:
a) rhesus serum albumin and cynomolgus serum albumin,
b) mouse serum albumin and rat serum albumin, or
c) rhesus serum albumin, cynomolgus serum albumin, mouse serum albumin, and rat serum albumin.
3 . A fusion polypeptide comprising
(a) polypeptide comprising a fibronectin type III tenth domain ( 10 Fn3) wherein the 10 Fn3 domain comprises AB, BC, CD, DE, EF, and FG loops, wherein the amino acid sequence of the CD loop of the wild-type 10 Fn3 domain set forth in SEQ ID NO: 1 is replaced by an amino acid sequence selected from the group consisting of SEQ ID NOs: 101-125, and wherein the polypeptide binds to human serum albumin, and to one or more of rhesus serum albumin, cynomolgus serum albumin, mouse serum albumin, and rat serum albumin; and
(b) at least one heterologous protein, wherein the at least one heterologous protein is linked to the polypeptide with (i) a polypeptide linker having a protease site or (ii) a biocompatible polymer;
wherein the polypeptide comprises an amino acid sequence at least 95% identical to the non-CD loop regions of SEQ ID NOs: 81, 88, 24-48, 50-74, 76-80, 82-87, 89-100, 185-209 and 236-260.
4 . A composition comprising the fusion polypeptide of claim 1 and a carrier.
5 . The fusion polypeptide of claim 1 , wherein the biocompatible polymer is a polymeric sugar.
6 . The fusion polypeptide of claim 1 , wherein the at least one heterologous protein further comprises a detectable moiety.
7 . The fusion polypeptide of claim 6 , wherein the detectable moiety is a radioisotope, a fluorescent compound, or a chemiluminescent compound.
8 . A fusion polypeptide comprising
(a) polypeptide comprising a fibronectin type III tenth domain ( 10 Fn3) wherein the 10 Fn3 domain comprises AB, BC, CD, DE, EF, and FG loops, wherein the amino acid sequence of the CD loop of the wild-type 10 Fn3 domain set forth in SEQ ID NO: 1 is replaced by an amino acid sequence selected from the group consisting of SEQ ID NOs: 101-125, and wherein the polypeptide binds to human serum albumin, and to one or more of rhesus serum albumin, cynomolgus serum albumin, mouse serum albumin, and rat serum albumin; and
(b) at least one heterologous protein, wherein the at least one heterologous protein is linked to the polypeptide with (i) a polypeptide linker having a protease site or (ii) a biocompatible polymer;
wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 24-30, 32-40, 42-48, 50-74, 76-100, 168-169, 185-191, 193-201, 203-209, 236-242, 243-252 and 254-260.
9 . The fusion polypeptide of claim 1 , wherein the biocompatible polymer is a polymeric sugar.
10 . The fusion polypeptide of claim 1 , wherein the heterologous protein is a therapeutic moiety.
11 . The fusion polypeptide of claim 1 , wherein the heterologous protein is a polypeptide comprising a 10 Fn3 domain.
12 . The fusion polypeptide of claim 11 , wherein the 10 Fn3 domain of the heterologous protein binds to a target protein other than serum albumin.
13 . The fusion polypeptide of claim 7 , wherein the radioisotope is 43 K, 52 Fe, 57 Co, 67 Cu, 67 Ga, 68 Ga, 123 I, 125 I, 131 I, 132 I, or 99 Tc.
14 . The fusion polypeptide of claim 7 wherein the fluorescent or chemiluminescent compound is fluorescein isothiocyanate, rhodamine, luciferin, alkaline phosphatase, beta-galactosidase or horseradish peroxidase.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jan 13, 2022
From: MITCHELL, TRACY S.; GOSSELIN, MICHAEL L.; LIPOVSEK, DASA; PARKER, REX; CAMPHAUSEN, RAY; DAVIS, JONATHAN; FABRIZIO, DAVID
To: BRISTOL-MYERS SQUIBB COMPANY
Reel/Frame 058651/0939 →
Continuity (4)
Division
15127166
Provisional Application
61968181
· Mar 20, 2014
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