Method and composition for enhancing transport across biological membranes

• US 6,495,663 B1
• Filed: 09/14/1999
• Issued: 12/17/2002
• Est. Priority Date: 05/21/1997
• Status: Expired due to Term

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Inventors: Wender, Paul A., Rothbard, Jonathan B.
• Primary Examiner(s): Low, Christopher S. F.
• Assistant Examiner(s): Lukton, David
• Application Number:
  US09/396,195
• Time in Patent Office: 1,190 Days
• Field of Search: 514/2, 514/12-17, 530/324-329
• US Class Current: 530/329
• CPC Class Codes: A61K 31/785 Polymers containing nitrogen
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• CPC Class Codes: A61K 47/645 Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• CPC Class Codes: C07D 305/14 condensed with carbocyclic rings or ring systems
• CPC Class Codes: C40B 40/00 Libraries per se, e.g. arrays, mixtures

Abstract

Methods and compositions for transporting drugs and macromolecules across biological membranes are disclosed. In one embodiment, the invention includes a method for enhancing transport of a selected compound across a biological membrane, wherein a biological membrane is contacted with a conjugate containing a biologically active agent that is covalently attached to a transport polymer. In one embodiment, the polymer consists of from 6 to 25 subunits, at least 50% of which contain a guanidino or amidino sidechain moiety. The polymer is effective to impart to the attached agent a rate of trans-membrane transport across a biological membrane that is greater than the rate of trans-membrane transport of the agent in non-conjugated form.

Claims

1. A conjugate for administration, to a mammalian subject, of an antimicrobial agent whose efficacy in non-conjugated form is limited by its solubility in aqueous liquid or its inability to cross biological membranes to manifest biological activity, said conjugate comprises a transport peptide covalently attached by a linker to the antimicrobial agent, said linker being cleaved in vivo to release the antimicrobial agent from the linker and the transport peptide, wherein the attached transport peptide:

(a) consists of from 6 to 25 amino acid residues, at least 50% of which contain a guanidino sidechain having the structure

or an amidino sidechain having the structure

wherein n is 2, 3, 4 or 5;

(b) contains at least 6 contiguous amino acid residues containing said guanidino or amidino sidechain; and

(c) increases the delivery of the antimicrobial agent across a biological membrane relative to the delivery of the antimicrobial agent in non-conjugated form.

2. The conjugate of claim 1, wherein at least 70% of the amino acid residues in the transport peptide contain said guanidino sidechain.

3. The conjugate of claim 1, wherein the transport peptide consists of from 7 to 20 amino acid residues.

4. The conjugate of claim 1, wherein each amino acid residue contains said guanidino sidechain.

5. The conjugate of claim 1, wherein all of the amino acid residues are arginine residues.

6. The conjugate of claim 5, wherein at least one of the arginine residues has a D-configuration.

7. The conjugate of claim 6, wherein all of the arginine residues have a D-configuration.

8. The conjugate of claim 1, wherein the transport peptide is attached at least one terminus to a flanking moiety that does not significantly affect the delivery of the antimicrobial agent across biological membranes.

9. The conjugate of claim 8, wherein the transport peptide has a first terminus and a second terminus, with the first terminus attached to a first flanking moiety and the second terminus conjugated to the antimicrobial agent through the linker.

10. The conjugate of claim 9, wherein a second flanking moiety is attached to the second terminus, such that the linker is attached to the second flanking moiety.

11. The conjugate of claim 9, where the first flanking moiety comprises one or more amino acid residues that do not contain said guanidino sidechain or said amidino sidechain.

12. The conjugate of claim 10, where the first flanking moiety and the second flanking moiety comprise one or more amino acid residues that do not contain said guanidino sidechain or said amidino sidechain.

13. The conjugate of claim 9, wherein the first flanking moiety is a blocking group effective to prevent ubiquitination in vivo.

14. The conjugate of claim 13, wherein the blocking group is an acetyl or benzyl group.

15. The conjugate of claim 1, wherein the linker contains a linkage that is chemically, enzymatically or photolytically cleaved in vivo.

16. The conjugate of claim 15, wherein the linkage is a carbamate, ester, thioether, disulfide or hydrazone linkage.

17. The conjugate of claim 1, wherein n is 3.

18. A pharmaceutical composition for administration, to a human subject, of an antimicrobial agent whose efficacy in non-conjugated form is limited by its aqueous solubility, said composition comprising the conjugate of claim 1 and a pharmaceutically acceptable carrier.