High molecular weight, post-translationally modified protein brushes

Patent metadata

Field	Value
Publication number	US-9963492-B2
Application number	US-201615155933-A
Country	US
Kind code	B2
Filing date	May 16, 2016
Priority date	May 14, 2015
Publication date	May 8, 2018
Grant date	May 8, 2018

Abstract

Disclosed are protein brushes that mimic mucin in physical and functional characteristics. The protein brushes have a variable number of tandem repeats similar to natural mucin and are modified at a number of their tyrosine residues to introduce brush substituents that mimic the hydrogel nature of mucin.

First claim

We claim:

1. A polypeptide comprising a plurality of tandem repeats of a sequence comprised by a mucin selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 11 and SEQ ID NO: 17, wherein about 20% to about 50% of the serine and threonine amino acids in the tandem repeats have been replaced by tyrosine residues.

2. The polypeptide of claim 1 , wherein the mucin is SEQ ID NO: 16 or SEQ ID NO: 17.

3. The polypeptide of claim 1 , wherein the N-terminal and C-terminal amino acids are cysteine.

4. The polypeptide of claim 1 , wherein the polypeptide is selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 11.

5. The polypeptide of claim 4 , wherein the polypeptide is selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8.

6. The polypeptide of claim 4 , wherein the polypeptide is SEQ ID NO: 2.

7. The polypeptide of claim 1 , wherein the number of tandem repeat sequences ranges from about 15 to about 70.

8. The polypeptide of claim 1 , wherein a plurality of the tyrosine residues have been modified to include a substituent selected from the group consisting of alkyl, alkynyl, aryl, amino, carboxyl, heteroaryl, nitro, sulfate, and polyethylene oxide polymer.

9. The polypeptide of claim 8 , wherein the plurality is at least 20% of the tyrosine residues.

10. A protein oligomer comprising at least two polypeptide units, wherein the polypeptide units comprise a plurality of tandem repeats of a sequence comprised by a mucin selected from the group consisting of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 11 and SEQ ID NO: 17; and about 20% to about 50% of the serine and threonine amino acids in the protein oligomer have been replaced by tyrosine residues.

11. The protein oligomer of claim 10 , wherein the mucin is SEQ ID NO: 16 or SEQ ID NO: 17.

12. The protein oligomer of claim 10 , wherein the N-terminal and C-terminal amino acids are cysteine.

13. The protein oligomer of claim 10 , wherein the polypeptide is selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 11.

14. The protein oligomer of claim 13 , wherein the polypeptide is selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8.

15. The protein oligomer of claim 13 , wherein the protein is SEQ ID NO: 2.

16. The protein oligomer of claim 10 , wherein the number of tandem repeat sequences ranges from about 15 to about 70.

17. The protein oligomer of claim 10 , wherein the at least two polypeptide units are linked together through a disulfide bond between the N-terminus of one protein and the C-terminus of the other protein.

18. The protein oligomer of claim 17 , wherein 2 to 30 polypeptide units are linked together using disulfide bonds.

19. The protein oligomer of claim 10 , wherein a plurality of the tyrosine residues have been modified to include a substituent selected from the group consisting of alkyl, alkynyl, aryl, amino, carboxyl, heteroaryl, nitro, sulfate, and polyethylene oxide polymer.

20. The protein oligomer of claim 19 , wherein the plurality is at least 20% of the tyrosine residues.

Assignees

• Seifried Brian M
• Olsen Bradley D
• Massachusetts Inst Technology

Inventors

• Seifried Brian M
• Olsen Bradley D

Classifications

• C07K14/4727 Primary

  Mucins, e.g. human intestinal mucin · CPC title

• C07K1/113

  without change of the primary structure · CPC title

• C07K14/47

  from mammals · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• C07K1/107

  by chemical modification of precursor peptides · CPC title