Engineered polypeptides for antigen delivery

Patent metadata

Field	Value
Publication number	US-10450352-B2
Application number	US-201716087215-A
Country	US
Kind code	B2
Filing date	Mar 23, 2017
Priority date	Mar 23, 2016
Publication date	Oct 22, 2019
Grant date	Oct 22, 2019

Abstract

Recombinant high affinity Invasin polypeptide are provided herein. Further provided are methods of delivering therapeutics such as vaccines by conjugation to the engineered recombinant Invasin polypeptides.

First claim

What is claimed is:

1. A recombinant polypeptide comprising an Invasin D5 domain coding sequence at least 80% identical to amino acid residues 887-986 of SEQ ID NO: 1, wherein the Invasin D5 domain coding sequence comprises an amino acid substitution at the position corresponding to Gly 909 and/or an amino acid substitution at the position corresponding to Ser 910.

2. The polypeptide of claim 1 , wherein the polypeptide comprises an amino acid substitution at the position corresponding to Gly 909 and an amino acid substitution at the position corresponding to Ser 910.

3. The polypeptide of claim 1 , wherein the polypeptide comprises an amino acid substitution for a residue having a positively charged side chain at the position corresponding to Gly 909.

4. The polypeptide of claim 3 , wherein the polypeptide comprises an Arg substitution at the position corresponding to Gly 909.

5. The polypeptide of claim 1 , wherein the polypeptide comprises an amino acid substitution for an aliphatic residue at the position corresponding to Ser 910.

6. The polypeptide of claim 5 , wherein the polypeptide comprises a Gly substitution at the position corresponding to Ser 910.

7. The polypeptide of claim 2 , wherein the polypeptide comprises an Arg substitution at the position corresponding to Gly 909 and a Gly substitution at the position corresponding to Ser 910.

8. The polypeptide of claim 7 , wherein the polypeptide comprises a 909 RGD 911 motif.

9. The polypeptide of claim 8 , wherein the 909 RGD 911 motif increases integrin binding affinity by at least 10 fold as compared to wild-type Invasin.

10. The polypeptide of claim 8 , wherein the 909 RGD 911 motif increases integrin binding affinity by at least 50 fold as compared to wild-type Invasin.

11. The polypeptide of claim 8 , wherein the 909 RGD 911 motif increases integrin binding affinity by at least 1000 fold as compared to wild-type Invasin.

12. The polypeptide of claim 1 , further comprising an Invasin D4 coding sequence at least 80% identical to amino acid residues 795-886 of SEQ ID NO: 1.

13. The polypeptide of claim 12 , wherein the Invasin D4 coding sequence comprises one or more of the following features:
(a) an amino acid substitution at the position corresponding to Phe 844;
(b) an amino acid substitution at the position corresponding to Tyr 878; or
(c) an amino acid substitution at the position corresponding to Tyr 885.

14. The polypeptide of claim 12 , wherein the polypeptide comprises an amino acid substitution for a residue having a hydrophobic side chain at the position corresponding to Phe 844.

15. The polypeptide of claim 14 , wherein the polypeptide comprises a Tyr substitution at the position corresponding to Phe 844.

16. A composition comprising a polypeptide of claim 1 in a pharmaceutically acceptable carrier.

17. A method for the delivery of a therapeutic agent to a subject comprising administering a therapeutic agent and a polypeptide of claim 1 to the subject.

18. A method of eliciting an immune response in a subject comprising administering at least one antigen and an Invasin polypeptide according to claim 1 to the subject.

19. The method of claim 18 , wherein the antigen is associated with a disease.

20. The method of claim 19 , wherein the disease is an infectious diseases, autoimmune disease, allergy, or cancer.

Assignees

• Univ Texas

Inventors

• Maynard Jennifer
• Wang Xianzhe
• Khan Tarik Ali

Classifications

• C07K14/47

  from mammals · CPC title

• C07K14/24 Primary

  from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia · CPC title

• A61K35/66

  Microorganisms or materials therefrom (fungi, yeasts or candida A61K36/06) · CPC title

• A61K47/66

  the modifying agent being a pre-targeting system involving a peptide or protein for targeting specific cells · CPC title

• A61K2039/6068

  Other bacterial proteins, e.g. OMP · CPC title