Fungal-specific metalloproteases and uses thereof

Patent metadata

Field	Value
Publication number	US-9493760-B2
Application number	US-201214342857-A
Country	US
Kind code	B2
Filing date	Sep 7, 2012
Priority date	Sep 8, 2011
Publication date	Nov 15, 2016
Grant date	Nov 15, 2016

Abstract

The present invention relates to methods of reducing, delaying, preventing and/or inhibiting the progression of a Cryptococcus infection into the central nervous system (CNS) of a subject by inhibiting the activity of a M36 fungalysin metalloprotease (e.g., MPR1) secreted by the Cryptococcus . The invention further provides methods of increasing, promoting and/or enhancing delivery of a therapeutic agent across the blood-brain-barrier, comprising systemically administering the therapeutic agent in conjunction with a M36 fungalysin metalloprotease (e.g., MPR1), or an enzymatically active fragment thereof.

First claim

What is claimed is:

1. A composition comprising a nanoparticle or a liposome attached to a M36 fungalysin metalloprotease fragment having at least 95% sequence identity to residues 446-722 of SEQ ID NO:1, wherein the fragment retains protease activity and can facilitate, increase and/or enhance delivery of the nanoparticle or the liposome across the blood-brain-barrier, and wherein a therapeutic agent is encapsulated within the nanoparticle or the liposome.

2. The composition of claim 1 , wherein the M36 fungalysin metalloprotease fragment does not comprise a fungalvsin/thermolvsin propeptide (FTP) domain.

3. The composition of claim 1 , wherein the M36 fungalysin metalloprotease fragment does not comprise a N-terminal signal peptide and a propeptide FTP domain.

4. The composition of claim 1 , wherein the M36 fungalysin metalloprotease fragment does not comprise N-terminal amino acid residues 1-300 of SEQ ID NO:1.

5. The composition of claim 1 , wherein the M36 fungalysin metalloprotease fragment does not comprise N-terminal amino acid residues 1-375 of SEQ ID NO:1.

6. The composition of claim 1 , wherein the M36 fungalysin metalloprotease fragment is conjugated or covalently bound to the nanoparticle or liposome.

7. The composition of claim 1 , wherein the composition is formulated in a pharmaceutically acceptable carrier.

8. The composition of claim 1 , wherein the nanoparticle is a silicon nanoparticle.

9. The composition of claim 1 , wherein the nanoparticle is selected from the group consisting of silicon (Si) nanoparticles, polylactide-co-glycolide nanoparticles, polyethyleneimine (PEI)-As(2)O(3)/Mn(0.5)Zn(0.5)Fe(2)O(4) magnetic nanoliposomes, redox-responsive poly(ethylene glycol)-b-poly(lactic acid) (MPEG-SS-PLA) nanoparticles, Thiolated Pluronic (Plu-SH) nanoparticles, mesoporous silica nanoparticles (MSNs) and biocompatible nanomicelles comprised of cholic acid, lysine and polyethylene glycol (PEG).

10. The composition of claim 1 , wherein the nanoparticle is comprised of a material selected from the group consisting of Poly(lactic acid-co-glycolic acid) (PLGA), biodegradable poly(L lactic acid) (PLLA), PEG-based hydrogels, and mixtures thereof.

11. The composition of claim 1 , wherein the M36 fungalysin metalloprotease fragment is integrated into the nanoparticle or the liposome.

12. The composition of claim 1 , wherein the M36 fungalysin metalloprotease fragment does not comprise a N-terminal signal peptide.

13. A method of increasing, promoting and/or enhancing delivery of a therapeutic agent across the blood-brain-barrier, comprising systemically administering to a subject in need thereof a composition comprising a nanoparticle or a liposome attached to a M36 fungalysin metalloprotease fragment having at least 95% sequence identity to residues 446-722 of SEQ ID NO:1, wherein the fragment retains protease activity and can facilitate, increase and/or enhance delivery of the therapeutic agent across the blood-brain-barrier, and wherein the therapeutic agent is encapsulated within the nanoparticle or the liposome.

14. The method of claim 13 , wherein the M36 fungalysin metalloprotease fragment does not comprise a fungalysin/thermolysin propeptide (FTP) domain.

15. The method of claim 13 , wherein the M36 fungalysin metalloprotease fragment, does not comprise a N-terminal signal peptide and propeptide FTP domain.

16. The method of claim 13 , wherein the M36 fungalysin metalloprotease fragment does not comprise N-terminal amino acid residues 1-300 of SEQ ID NO:1.

17. The method of claim 13 , wherein the M36 fungalysin metalloprotease fragment does not comprise N-terminal amino acid residues 1-375 of SEQ ID NO:1.

18. The method of claim 13 , wherein the therapeutic agent is a cell, small organic molecule, small inorganic molecule, a peptide, a polypeptide or a nucleic acid.

19. The method of claim 13 , wherein the therapeutic agent is effective in the treatment, mitigation or prevention of a cancer of the central nervous system or a neurodegenerative disease.

Assignees

• Gelli Angela C
• Univ California

Inventors

• Gelli Angela C

Classifications

• C12N9/58 Primary

  derived from fungi · CPC title

• A61P31/12

  Antivirals · CPC title

• C12Y203/01

  transferring groups other than amino-acyl groups (2.3.1) · CPC title

• C07K16/40 Primary

  against enzymes · CPC title

• C12N9/1029

  transferring groups other than amino-acyl groups (2.3.1) · CPC title