Proteins specific for pyoverdine and pyochelin

Patent metadata

Field	Value
Publication number	US-10065998-B2
Application number	US-201715726577-A
Country	US
Kind code	B2
Filing date	Oct 6, 2017
Priority date	Feb 18, 2015
Publication date	Sep 4, 2018
Grant date	Sep 4, 2018

Abstract

The present disclosure provides hNGAL muteins that bind a pyoverdine family member or pyochelin and can be used in various application including pharmaceutical applications, for example, to inhibit or reduce growth of P. aeruginosa . The present disclosure also concerns methods of making one or more pyoverdine- or pyochelin-binding muteins described herein as well as compositions comprising one or more of such muteins. The present disclosure further relates to nucleic acid molecules encoding such muteins and to methods for generation of such muteins and nucleic acid molecules. In addition, the application discloses therapeutic and/or diagnostic uses of these muteins as well as compositions comprising one or more of such muteins.

First claim

The invention claimed is:

1. A human neutrophil gelatinase-associated lipocalin (hNGAL) mutein polypeptide having binding specificity for pyoverdine type III, wherein the hNGAL mutein comprises 10 or more mutations at positions 36, 40, 41, 42, 49, 52, 65, 68, 70, 73, 77, 79, 81, 87, 96, 100, 103, 105, 106, 127, and 134 of SEQ ID NO: 1.

2. The hNGAL mutein polypeptide of claim 1 , wherein the hNGAL mutein comprises ten or more of the following mutations of SEQ ID NO: 1: L36E, A40L, I41A, L42R, Q49M, Y52M, N65D, S68E, L70R, K73H, D77K, W79D, R81V, C87S, N96L, Y100D, L103Q, S105P, Y106E, S127V, K134W.

3. The hNGAL mutein polypeptide of claim 1 , wherein the hNGAL mutein comprises SEQ ID NO: 53.

4. The hNGAL mutein polypeptide of claim 1 , wherein the hNGAL mutein has binding specificity for pyoverdine type III succinyl, pyoverdine type III succinamid, and/or pyoverdine type III α-ketoglutaryl.

5. The hNGAL mutein polypeptide of claim 1 , wherein the hNGAL mutein is capable of binding pyoverdine type III complexed with iron with a K D of about 200 nM or lower.

6. The hNGAL mutein polypeptide of claim 1 , wherein the hNGAL mutein comprises 12 or more mutations at positions 36, 40, 41, 42, 49, 52, 65, 68, 70, 73, 77, 79, 81, 87, 96, 100, 103, 105, 106, 127, and 134 of SEQ ID NO: 1.

7. A human neutrophil gelatinase-associated lipocalin (hNGAL) mutein polypeptide having binding specificity for pyoverdine type III, wherein the hNGAL mutein comprises 15 or more mutations at positions 36, 40, 41, 42, 49, 52, 65, 68, 70, 73, 77, 79, 81, 87, 96, 100, 103, 105, 106, 127, and 134.

8. The hNGAL mutein polypeptide of claim 7 , wherein the hNGAL mutein comprises 15 or more of the following mutations of SEQ ID NO: 1: L36E, A40L, I41A, L42R, Q49M, Y52M, N65D, S68E, L70R, K73H, D77K, W79D, R81V, C87S, N96L, Y100D, L103Q, S105P, Y106E, S127V, K134W.

9. The hNGAL mutein polypeptide of claim 7 , wherein the hNGAL mutein comprises SEQ ID NO: 53.

10. The hNGAL mutein polypeptide of claim 7 , wherein the hNGAL mutein has binding specificity for pyoverdine type III succinyl, pyoverdine type III succinamid, and/or pyoverdine type III α-ketoglutaryl.

11. The hNGAL mutein polypeptide of claim 7 , wherein the hNGAL mutein is capable of binding pyoverdine type III complexed with iron with a K D of about 200 nM or lower.

12. A human neutrophil gelatinase-associated lipocalin (hNGAL) mutein polypeptide having binding specificity for pyoverdine type III, wherein the hNGAL mutein comprises SEQ ID NO: 53.

13. The hNGAL mutein polypeptide of claim 12 , wherein the hNGAL mutein consists of SEQ ID NO: 53.

14. The hNGAL mutein polypeptide of claim 12 , wherein the hNGAL mutein has binding specificity for pyoverdine type III succinyl, pyoverdine type III succinamid, and/or pyoverdine type III α-ketoglutaryl.

15. The hNGAL mutein polypeptide of claim 12 , wherein the hNGAL mutein is capable of binding pyoverdine type III complexed with iron with a K D of about 200 nM or lower.

16. A composition comprising the hNGAL mutein polypeptide of claim 1 , and at least one pharmaceutically acceptable adjuvant, diluent, or carrier.

17. A composition comprising the hNGAL mutein polypeptide of claim 7 , and at least one pharmaceutically acceptable adjuvant, diluent, or carrier.

18. A composition comprising the hNGAL mutein polypeptide of claim 12 , and at least one pharmaceutically acceptable adjuvant, diluent, or carrier.

Assignees

• Sanofi Sa

Inventors

• Corvey Carsten
• Stump Heike
• Kruip Jochen
• Calandra Bernhard
• Rey Astrid
• Karst Nathalie
• Mourez Michael
• Fraisse Laurent
• Rothe Christine
• Allersdorfer Andrea
• Wiedenmann Alexander
• Hinner Marlon
• Lunde Bradley
• Jensen Kristian
• Hülsmeyer Martin

Classifications

• A61P31/04

  Antibacterial agents · CPC title

• A61P5/00

  Drugs for disorders of the endocrine system · CPC title

• A61K38/005 Primary

  Enzyme inhibitors (protease inhibitors A61K38/55) · CPC title

• A61K38/012

  from animals · CPC title

• C07K14/4703 Primary

  Inhibitors; Suppressors · CPC title