Multivalent heteromultimer scaffold design and constructs

Patent metadata

Field	Value
Publication number	US-10358479-B2
Application number	US-201615001078-A
Country	US
Kind code	B2
Filing date	Jan 19, 2016
Priority date	Jul 13, 2012
Publication date	Jul 23, 2019
Grant date	Jul 23, 2019

Abstract

Provided herein are multifunctional heteromultimer proteins. In specific embodiments is a heteromultimer comprising: at least two polypeptide constructs, each polypeptide construct comprising at least one cargo polypeptide attached to a transporter polypeptide, said transporter polypeptides derived from a monomeric native protein such that said monomeric constructs associate to form the heteromultimer and said transporter polypeptides associate to form a quasi-native structure of the monomeric native protein or analog thereof. These therapeutically novel molecules encompass heteromultimers comprising constructs that function as scaffolds for the conjugation or fusion of therapeutic molecular entities (cargo polypeptides) resulting in the creation of bispecific or multivalent molecular species. Provided herein is a method for creation of bispecific or multivalent molecular species.

First claim

What is claimed:

1. A heteromultimer comprising:
a first polypeptide construct comprising (i) a first transporter polypeptide comprising a first segment of albumin, and (ii) at least one first cargo molecule; and
a second polypeptide construct comprising (iii) a second transporter polypeptide comprising a second segment of albumin,
wherein a) said first segment of albumin and said second segment of albumin are obtained by segmentation of an albumin polypeptide at a segmentation site to delete zero to 3 amino acid residues at the segmentation site, b) said first transporter polypeptide and said second transporter polypeptide are different from each other, and c) said first transporter polypeptide and said second transporter polypeptide self-assemble to form a quasi-native structure of albumin, and
wherein:

a) said first transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:39 without the signal peptide sequence, and wherein said second transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:40 without the signal peptide sequence;
b) said first transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:41 without the signal peptide sequence, and wherein said second transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:42 without the signal peptide sequence;
c. said first transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:43 without the signal peptide sequence, and wherein said second transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:44 without the signal peptide sequence;
d) said first transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:45 without the signal peptide sequence, and wherein said second transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:46 without the signal peptide sequence;
e) said first transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:47 without the signal peptide sequence, and wherein said second transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:48 without the signal peptide sequence;
f) said first transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:49 without the signal peptide sequence, and wherein said second transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:50 without the signal peptide sequence;
g) said first transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:51 without the signal peptide sequence, and wherein said second transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:52 without the signal peptide sequence; or
h) said first transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:53 without the signal peptide sequence, and wherein said second transporter polypeptide comprises a sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO:54 without the signal peptide sequence.

2. The heteromultimer of claim 1 , wherein the activity of the cargo molecule is altered on fusion to the transporter polypeptide relative to its unfused format, or wherein the activity of the cargo molecule is attenuated on fusion to the transporter polypeptide relative to its unfused format.

3. The heteromultimer of claim 1 , wherein said heteromultimer binds to FcRn.

4. A composition comprising the heteromultimer of claim 1 and a pharmaceutically acceptable carrier.

5. One or more nucleic acids encoding the heteromultimer according to claim 1 .

6. One or more vectors comprising the one or more nucleic acids according to claim 5 .

7. A host cell comprising the one or more nucleic acids according to claim 5 or the one or more vectors according to claim 6 .

8. The heteromultimer of claim 1 , wherein said second polypeptide construct further comprises at least one second cargo molecule.

9. The heteromultimer of claim 1 , wherein
a) said first transporter polypeptide comprises the sequence set forth in SEQ ID NO:39 without the signal peptide sequence, and wherein said second transporter polypeptide comprises the sequence set forth in SEQ ID NO:40 without the signal peptide sequence;
b) said first transporter polypeptide comprises the sequence set forth in SEQ ID NO:41 without the signal peptide sequence, and wherein said second transporter polypeptide comprises the sequence set forth in SEQ ID NO:42 without the signal peptide sequence;
c. said first transporter polypeptide comprises the sequence set forth in SEQ ID NO:43 without the signal peptide sequence, and wherein said second transporter polypeptide comprises the sequence set forth in SEQ ID NO:44 without the signal peptide sequence;
d) said first transporter polypeptide comprises the sequence set forth in SEQ ID NO:45 without the signal peptide sequence, and wherein said second transporter polypeptide comprises the sequence set forth in SEQ ID NO:46 without the signal peptide sequence;
e) said first transporter polypeptide comprises the sequence set forth in SEQ ID NO:47 without the signal peptide sequence, and wherein said second transporter polypeptide comprises the sequence set forth in SEQ ID NO:48 without the signal peptide sequence;
f) said first transporter polypeptide comprises the sequence set forth in SEQ ID NO:49 without the signal peptide sequence, and wherein said second transporter polypeptide comprises the sequence set forth in SEQ ID NO:50 without the signal peptide sequence;
g) said first transporter polypeptide comprises the sequence set forth in SEQ ID NO:51 without the signal peptide sequence, and wherein said second transporter polypeptide comprises the sequence set forth in SEQ ID NO:52 without the signal peptide sequence; or
h) said first transporter polypeptide comprises the sequence set forth in SEQ ID NO:53 without the signal peptide sequence, and wherein said second transporter polypeptide comprises the sequence set forth in SEQ ID NO:54 without the signal peptide sequence.

10. The heteromultimer of claim 1 , wherein the first transporter polypeptide or the second transporter polypeptide further comprises a mutation that improves stability or half-life of the heteromultimer.

11. The heteromultimer of claim 8 , wherein the at least one first cargo molecule or at least one second cargo molecule is a therapeutic agent, a toxin, a natural product, a prodrug, a cargo polypeptide, or a nucleic acid.

12. The heteromultimer of claim 8 , wherein the first polypeptide construct comprises one first cargo molecule or two first cargo molecules wherein the first cargo molecules are cargo polypeptides, and/or the second polypeptide construct comprises one second cargo molecule or two second cargo molecules wherein the second cargo molecules are cargo polypeptides.

13. The heteromultimer of claim 12 , wherein at least one of the first cargo polypeptides or at least one of the second cargo

Assignees

• Zymeworks Inc

Inventors

• Dixit Surjit Bhimarao
• D'Angelo Igor Edmondo Paolo
• Sanches Mario
• Ng Gordon Yiu Kon

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• C07K2317/35

  Valency · CPC title

• C07K16/2809

  against the T-cell receptor (TcR)-CD3 complex · CPC title

• C07K16/40

  against enzymes · CPC title