NOVEL PH -SWITCHABLE PEPTIDES FOR MEMBRANE INSERTION AND PORE FORMATION

• US 20150374844A1
• Filed: 02/10/2014
• Published: 12/31/2015
• Est. Priority Date: 02/15/2013
• Status: Abandoned Application

• Current Assignee: Sirna Therapeutics, Inc. (Alnylam Pharmaceuticals Incorporated), University of Pennsylvania
• Original Assignee: Sirna Therapeutics, Inc. (Alnylam Pharmaceuticals Incorporated), University of Pennsylvania
• Inventors: DEGRADO, William, TELLERS, David M., GRIGORYAN, Gevorg, JADHAV, Vasant, ZHANG, Yao
• Application Number:
  US14/768,207
• Publication Number:
  US 20150374844A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 38/00 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 Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: C07K 14/00 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 14/001 by chemical synthesis

Abstract

Disclosed herein is a pH-switchable pore formation (PSPF) peptide comprising one or more amino acids in peptide sequence whose charge state and hydrophobicity are pH-dependent, wherein the peptide can bind to a biological membrane upon contact and form pores on the membrane at pH of less than about 7, and wherein the peptide forms substantially no pores on the biological membrane at pH of greater than about 7. Also disclosed is a modular composition comprising: a) one or more PSPF peptides, which may be the same or different; b) a single stranded or double stranded oligonucleotide; and c) one or more linkers, which may be the same or different.

Claims

1. A pH-switchable pore formation (PSPF) peptide comprising one or more amino acids in peptide sequence whose charge state and hydrophobicity are pH-dependent, wherein the peptide can bind to a biological membrane upon contact and form pores on the membrane at pH of less than about 7, and wherein the peptide forms substantially no pores on the biological membrane at pH of greater than about 7.

2. The PSPF peptide of claim 1, wherein the peptide can bind to a biological membrane upon contact and form pores on the membrane at pH of less than 6.5, and wherein the peptide forms substantially no pores on the biological membrane at pH of greater than 7.0.

3. The PSPF peptide of claim 1, wherein the peptide can bind to a biological membrane upon contact and form pores on the membrane at pH of about 5.5, and wherein the peptide forms substantially no pores on the biological membrane at pH of about 7.4.

4. The PSPF peptide of claim 1, wherein the peptide is water soluble at pH of greater than about 7.

5. The PSPF peptide of claim 1, wherein the amino acid is selected from the group consisting of Asp, Glu and His.

6. The PSPF peptide of claim 1, wherein the pores formed on the membrane serve as channels for transport of appropriately-sized target;

and wherein uptake of the peptide by endocytosis allows endosomal escape of material present in the extracellular environment into the cell.

7. The PSPF peptide of claim 1, wherein the peptide is selected from peptides of SEQ. ID No. 1-24.

8. The PSPF peptide of claim 7, wherein the peptide is selected from peptides of SEQ. ID No. 4, 8 and 12.

9. The PSPF peptide of claim 1 having a heptad repeat structure as shown in FIG. 2, wherein positions “

b”

in water is an amino acid selected from the group consisting of Ser and Thr.

10. The PSPF peptide of claim 1 having a heptad repeat structure as shown in FIG. 2, wherein position “

c”

in water is an amino acid selected from the group consisting of Asp, Glu and His.

11. The PSPF peptide of claim 10, wherein position “

c”

in water is Glu.

12. The PSPF peptide of claim 1 having a heptad repeat structure as shown in FIG. 2, wherein each of positions “

e” and

“

g”

is an amino acid independently selected from the group consisting of Ala, Gly, Ser and Thr.

13. A modular composition comprising:

a) one or more PSPF peptides of claim 1, which may be the same or different;

b) a single stranded or double stranded oligonucleotide;

c) optionally one or more linkers, which may be the same or different;

d) optionally one or more targeting ligands, which may be the same or different;

e) optionally one or more other peptides; and

f) optionally one or more lipids, which may be the same or different.

14. A modular composition comprising:

a) one or more PSPF peptides of claim 1, which may be the same or different;

b) a single stranded or double stranded oligonucleotide; and

c) one or more linkers, which may be the same or different.

15. The modular composition of claim 14, wherein the oligonucleotide is a double stranded siRNA;

and wherein each PSPF peptide is independently selected from peptides of SEQ. ID No. 1-24.

16. The modular composition of claim 14, further comprising:

d) one or more ligands, which may be the same or different.

17. The modular composition of claim 16, wherein each ligand is independently selected from the group consisting of D-galactose, N-acetyl-D-galactosamine (GalNAc), GalNAc2, and GalNAc3, GalNAc4, cholesterol, folate, and derivatives thereof.

18. The modular composition of claim 16 comprising:

a) 1-4 PSPF peptides independently selected from SEQ ID No. 1-24;

b) a double stranded siRNA;

c) 1-4 linkers independently selected from Table 4, which may be the same or different; and

d) 1-4 GalNAc ligands, which may be the same or different;

wherein the GalNAc ligands and/or the peptides are attached to the siRNA optionally via linkers.

19. A pharmaceutical composition comprising the PSPF peptide of claim 1 and a pharmaceutically acceptable excipient.

20. A pharmaceutical composition comprising the modular composition of claim 9 and a pharmaceutically acceptable excipient.