COMPOSITIONS FOR ENHANCING TRANSPORT OF MOLECULES INTO CELLS

• US 20090082547A1
• Filed: 11/05/2008
• Published: 03/26/2009
• Est. Priority Date: 04/29/2003
• Status: Active Grant

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: Nelson, Michelle H., Kroeker, Andrew D., Iversen, Patrick L., Moulton, Hong M., Stein, David A.
• Granted Patent: US 9,572,899 B2
• Time in Patent Office: Days
• US Class Current: 530/322
• 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: A61K 49/0043 Fluorescein, used in vivo
• CPC Class Codes: A61K 49/0054 Macromolecular compounds, i.e. oligomers, polymers, dendrimers
• CPC Class Codes: A61K 49/0056 Peptides, proteins, polyamino acids
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/314 Phosphoramidates
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/3513 Protein; Peptide
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific

Abstract

Compositions and methods for enhancing delivery of molecules, e.g. biological agents, into cells are described. The composition is a conjugate of the biological agent, preferably a nucleic acid analog having a substantially uncharged backbone, covalently linked to a peptide transporter moiety as described. Conjugation of the peptide transporter to a substantially uncharged nucleic acid analog, such as a morpholino oligomer, is also shown to enhance binding of the oligomer to its target sequence and enhance antisense activity.

Claims

1. A peptide-oligomer conjugate, comprising:

a carrier peptide attached to a nucleic acid analog having a substantially uncharged backbone and a targeting base sequence,

wherein the carrier peptide has the formula (ArgYArg)₄ or (ArgArgY)₄, where each Y is —

C(O)—

(CH₂)_n—

NH, where n is 2 to 7.

2. The conjugate of claim 1, wherein the carrier peptide has the formula (ArgYArg)₄.

3. The conjugate of claim 2, wherein the nucleic acid analog is attached to the peptide via a linker comprising a further Y subunit.

4. The conjugate of claim 3, wherein the linker comprises a β

-alamine or 6-aminohexanoic acid subunit.

5. The conjugate of claim 2, wherein the nucleic acid analog is attached to the peptide via an Ahx-β

Ala (6-aminohexanoic acid-β

-alamine) linker.

6. The conjugate of claim 1, wherein the nucleic acid analog is a morpholino oligomer, comprising morpholino subunits linked by phosphorus-containing linkages between the morpholino nitrogen of one subunit and an exocyclic carbon at the morpholino 3-position of an adjacent subunit.

7. The conjugate of claim 6, where the morpholino subunits are joined by intersubunit linkages in accordance with the structure:

8. The conjugate of claim 1, wherein the targeting base sequence of the nucleic acid analog is targeted to an antisense target in a polynucleotide, selected from a splice site in a pre-mRNA, a translation start site in an mRNA, and a cis-acting element in a viral genome.