Hydrophobic modified peptides and their use for liver specific targeting

Patent metadata

Field	Value
Publication number	US-10513539-B2
Application number	US-201213984694-A
Country	US
Kind code	B2
Filing date	Feb 10, 2012
Priority date	Feb 10, 2011
Publication date	Dec 24, 2019
Grant date	Dec 24, 2019

Abstract

The present invention relates to hydrophobic modified peptides for the specific delivery of compounds to the liver, preferably to hepatocytes, in vitro as well as in vivo. The present invention relates to pharmaceutical compositions comprising said hydrophobic modified peptide(s) and the compound(s) to be specifically delivered to the liver. The present invention furthermore relates to the use of the inventive hydrophobic modified peptides as well as to a method for the prevention and/or treatment of liver diseases or disorders.

First claim

The invention claimed is:

1. A hydrophobic modified peptide of the formula
[X—P—Y—R o ]A p5

wherein P is a peptide having the amino acid sequence NPLGFXaaP (SEQ ID NO: 1), wherein Xaa is F;
X is an amino acid sequence having a length of m amino acids,
wherein one or more of the amino acids of X carry one or more groups for hydrophobic modification selected from acylation comprising myristoyl (C 14) or stearoyl (C 18);
m is at least 4;
Y is an amino sequence having a length of n amino acids, wherein n is 0 or at least 1;
m+n≥11
R is a C-terminal modification of said hydrophobic modified peptide with an amide and o is 0 or at least 1,
A is an anchor group selected from ester, ether, disulfide, amide, thiol, thioester, p is 0 or at least 1,
wherein one or more drug(s) is/are coupled or linked to one or more amino acid(s) of X.

2. The hydrophobic modified peptide according to claim 1 , wherein m is 4 to 19 and/or n is 0 to 78.

3. The hydrophobic modified peptide according to claim 1 , wherein the one or more drug(s) is/are linked to said peptide via a linker or spacer.

4. The hydrophobic modified peptide according to claim 3 , wherein the linker or spacer is cleaved by a hepatocellular proteolytic enzyme.

5. The hydrophobic modified peptide according to claim 4 , wherein the enzyme is selected from the group consisting of cytochrome P450, a proteases of the endocytic pathway, a lyase of the endocytic pathway, matrix-metallo-protease 1 (MMP1), matrix-metallo-protease 2 (MMP2), matrix-metallo-protease 7 (MMP7), matrix-metallo-protease 9 (MMP9), matrix-metallo-protease 12 (MMP12), and matrix-metallo-protease 7 (MMP7).

6. The hydrophobic modified peptide according to claim 4 , wherein the linker or spacer comprises the peptide sequences GCHAK (SEQ ID NO: 19) or RPLALWRS (SEQ ID NO:20).

7. The hydrophobic modified peptide according to claim 1 , wherein one or more drug(s) is/are coupled to one or more amino acid(s) of X having an amino group in a side chain.

8. The hydrophobic modified peptide according to claim 7 , wherein the amino acid(s) having an amino group in a side chain is/are lysine.

9. The hydrophobic modified peptide according to claim 1 , wherein the one or more amino acid(s) having an amino group in a side chain are located at the N-terminus of X.

10. The hydrophobic modified peptide according to claim 9 , wherein 1 to 11 amino acids having an amino group in a side chain are located at the N-terminus of X.

11. The hydrophobic modified peptide according to claim 10 , wherein the drug(s) is/are coupled to one or more of the 1 to 11 amino acid(s) having an amino group in a side chain via an activated ester.

12. The hydrophobic modified peptide according to claim 1 , wherein the drug(s) is/are coupled to the amino acid(s) of X by using one or more methods selected from
formation of amides by the reaction of an amine and activated carboxylic acids,
disulfide linkage using two thiols or one thiol that specifically reacts with pyridyl disulfides;
thioether formation using maleimides or haloacetyls and a thiol component;
amidine formation using an imidoester and an amine;
hydrazide linkage using carbonyls and hydrazides;
amine linkage using carbonyls and amines under reductive conditions;
triazol formation using nitriles and azides;
thiourea formation using isothiocyanates and amines;
formation of esters by the reaction of an alcohol and activated carboxylic acids; and
formation of ethers by the reaction of an alcohol and alkyl halides.

13. The hydrophobic modified peptide according to claim 1 , wherein the hydrophobic modification is by acylation with myristoyl (C 14).

14. The hydrophobic modified peptide according to claim 1 , wherein the hydrophobic modification is by acylation with stearoyl (C 18).

15. The hydrophobic modified peptide according to claim 1 , wherein said peptide comprises of SEQ ID NOs: 2 to 13 with an amino acid sequence of different viral species, strains or subtypes selected from the genotypes consisting of HBC, wooly monkey HBV, or variants thereof.

16. The hydrophobic modified peptide according to claim 1 , wherein the drug(s) coupled or linked to one or more amino acid(s) of X is a combination of two or more drugs.

17. The hydrophobic modified peptide according to claim 16 , wherein the drug is selected from doxorubicin, a gyrase inhibitors of the fluoroquinilone class, primaquine, clindamycin, azithromycin, ciprofloxacin, doxocycline, atovaquone, rifampicin, and dapsone.

18. The hydrophobic modified peptide according to claim 16 , wherein the drug is selected from doxorubicin, oxaliplatin, irinotecan, fluorouracil, -bortezomib, sorafenib, botezomib, erlotinib, interferon-α, cisplatin, and lencovorin.

19. The hydrophobic modified peptide according to claim 17 , wherein the drug is selected from the group consisting of primaquine, doxorubicin and a gyrase inhibitors.

20. The hydrophobic modified peptide according to claim 18 , wherein the combination of two or more drugs is selected from a combination of bortezumib and doxorubicin; sorafenib and doxorubicin; botezomib and sorafenib; erlotinib and fluorouracil; erlotinib, fluorouracil and interferon-α; cisplatin and doxorubicin; cisplatin, doxorubicin and erlotinib; irinotecan and fluorouracil; irinotecan and erotinib; oxaliplatin and fluorouracil; and FOLFOX (oxaliplatin, fluorouracil, lencovorin).

21. The hydrophobic modified peptide according to claim 1 , which is selected from the group consisting of
stearoyl-[K(primaquine)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
stearoyl-[K(GCHAK(SEQ ID NO: 19) primaquine)]-NLSTSNPLGFFPDHQLDP (SEQ ID NO:21), myristoyl-[K(Doxocyclin)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO: 15),
myristoyl-[K(GCHAK(SEQ ID NO: 19) Doxocylin)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
myristoyl-[K(Penicillin)]3-NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
myristoyl-[K(GCHAK(SEQ ID NO: 19)-Penicillin)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
myristoyl-[K(Cyclosporine)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
myristoyl-[K(GCHAK(SEQ ID NO: 19)-Cyclosporine)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
myristoyl-[K(sirolimus)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
myristoyl-[K(GCHAK sirolimus)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
stearoyl-[K(RPLALWRS(SEQ ID NO: 20) bortezomib)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
stearoyl-[K(sorafenib)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
stearoyl-[K(RPLALWRS(SEQ ID NO: 20) sorafenib)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
stearoyl-[K(Thiazolidinedione)] 3 -NLSTSNPLGFFPDHQLDP (SEQ ID NO:15),
myristoyl-[K-Levofloxacin]-NLSTSNPLGFFPDHQLDP (SEQ ID NO:21), and
myristoyl-[K-Doxorubicin]-NLSTSNPLGFFPDHQLDP (SEQ ID NO:21).

22. A pharmaceutical composition comprising: at least one hydrophobic modified peptide according to claim 1 and a pharmaceutically acceptable carrier and/or excipient.

23. The hydrophobic modified peptide according to claim 1 or a pharmaceutical composition comprising said peptide for use in the specific delivery of a drug to the liver.

24. The hydrophobic modified peptide or pharmaceutical composition according to claim 23 for use in the treatment of a liver disease or disorder selected from the group consisting of virus-induced hepatitis or cirrhosis, malaria, and a liver tumor.

25. The hydrophobic modified peptide or pharmaceutical composition according to claim 23 , wherein the hydrophobic modified peptide is administered to a patient in a dosage rangi

Assignees

• Mier Walter
• Urban Stephan
• Mehrle Stefan
• Haberkorn Uwe
• Univ Heidelberg Ruprecht Karls

Inventors

• Mier Walter
• Urban Stephan
• Mehrle Stefan
• Haberkorn Uwe

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P33/12

  Schistosomicides · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P33/06

  Antimalarials · CPC title