Method for enhancing transmembrane transport of exogenous molecules

• US 5,416,016 A
• Filed: 03/13/1992
• Issued: 05/16/1995
• Est. Priority Date: 04/03/1989
• Status: Expired due to Term

• Current Assignee: Purdue Research Foundation (The Trustees of Purdue University (d/b/a Purdue University))
• Original Assignee: Purdue Research Foundation (The Trustees of Purdue University (d/b/a Purdue University))
• Inventors: Low, Philip S., Heinstein, Peter F., Horn, Mark A.
• Primary Examiner(s): Elliott, George C.
• Application Number:
  US07/851,544
• Time in Patent Office: 1,159 Days
• Field of Search: 435/240.1, 435/240.2, 435/243, 435/240.4, 435/172.3, 514/2, 514/44, 935/52, 424/450
• US Class Current: 435/375
• CPC Class Codes: A61K 47/551 one of the codrug's components being a vitamin, e.g. niacinamide, vitamin B3, cobalamin, vitamin B12, folate, vitamin A or retinoic acid
• CPC Class Codes: A61K 47/557 the modifying agent being biotin
• CPC Class Codes: A61K 47/665 the pre-targeting system, clearing therapy or rescue therapy involving biotin-(strept) avidin systems
• CPC Class Codes: A61K 49/001 Preparation for luminescence or biological staining
• CPC Class Codes: A61K 51/0497 conjugates with a carrier being an organic compounds
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07B 59/004 Acyclic, carbocyclic or heterocyclic compounds containing elements other than carbon, hydrogen, halogen, oxygen, nitrogen, sulfur, selenium or tellurium
• CPC Class Codes: C12N 15/64 General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
• CPC Class Codes: C12N 15/8206 by physical or chemical, i.e. non-biological, means, e.g. electroporation, PEG mediated
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12Q 1/68 involving nucleic acids

Abstract

A method is provided for enhancing transmembrane transport of exogenous molecules. The method comprises contacting a membrane of a living cell with a complex formed between said molecules and ligands selected from biotin, biotin analogs and other biotin receptor-binding ligands, and/or folic acid, folate analogs and other folate receptor-binding ligands to initiate receptor mediated transmembrane transport of the ligand complex. The method is used for the efficient delivery of peptides, proteins, nucleic acids and other compounds capable of modifying cell function into plant, animal, yeast, and bacterial cells.

Claims

1. A method for enhancing transport of an exogenous molecule across a membrane of a living cell, said method comprising the step of contacting the membrane with the exogenous molecule complexed with a ligand selected from the group consisting of folic acid, folate receptor-binding analogs of folic acid, and other folate receptor-binding ligands, for a time sufficient to permit transmembrane transport said ligand complex.

2. The method of claim 1 wherein the ligand is folic acid, folinic acid, pteropolyglutamic acid, folate receptor-binding pteridines and the deaza and dideaza analogs thereof.

3. The method of claim 1 wherein the living cell is a eukaryote.

4. The method of claim 3 wherein the living cell is an plant cell.

5. The method of claim 3 wherein the living cell is an animal cell.

6. The method of claim 5 wherein the living cell is a human cell.

7. The method of claim 5 wherein the living cell is selected from those lining the alimentary canal.

8. The method of claim 5 wherein the living cell forms part of tissue selected from the group consisting of dermis, epidermis, nasal mucosa, vaginal mucosa, placenta, eye, and the tissue comprising the blood/brain barrier.

9. The method of claim 3 wherein the exogenous molecule is a pharmaceutically active compound.

10. The method of claim 1 wherein the living cell is a prokaryote.

11. In a method for modifying cellular function by contacting a cell with a compound capable of modifying said function, the improvement which comprises contacting the cell with a complex of said compound with a ligand selected from the group consisting of folic acid, folate receptor-binding analogs of folic acid, and other folate receptor-binding ligands, for a time sufficient to permit transmembrane transport of said ligand complex.

12. The method of claim 11 wherein the ligand is folic acid, folinic acid, pteropolyglutamic acid, folate receptor-binding pteridines, and the deaza and dideaza analogs thereof.

13. The method of claim 11 wherein the compound is selected from proteins, oligopeptides, polypeptides, nucleic acids, and biologically functional analogs thereof.

14. A method for enhancing the introduction of nucleic acids and nucleic acid analogs into living cells which method comprises the step of contacting said cell with said nucleic acid, or said nucleic acid analog, complexed with a ligand selected from the group consisting of folic acid, folate receptor-binding analogs of folic acid, and other folate receptor-binding ligands, for a time sufficient to permit transmembrane transport said ligand complex.

15. The method of claim 14 wherein the nucleic acid or nucleic acid analog is substantially complementary to at least one endogenous nucleic acid in the living cell.

16. The method of claim 14 wherein the nucleic acid is incorporated into the heritable genome of the cell.

17. The method of claim 16 wherein the nucleic acid is a plasmid.

18. A method for potentiating the transport of an exogenous molecule contained within a liposome across a membrane of a living cell, the method comprising the step of contacting the membrane with a liposome containing the exogenous molecule, said liposome bearing ligands associated with its membrane contacting surface selected from the group consisting of folic acid, folate receptor-binding analogs of folic acid, and other folate receptor-binding ligands, for a time sufficient to permit transmembrane transport of said exogenous molecule contained within the liposome.

19. The method of claim 18 wherein the liposome comprises a phospholipid covalently bound to a folate receptor-binding ligand.

20. The method of claim 19 wherein the liposome comprises a phospholipid having a folate receptor-binding ligand covalently bound through the head group of said phospholipid.

21. The method of claim 19 wherein the folate receptor-binding ligand is covalently bound through a spacer arm or bridging molecule.

22. A liposome composition comprising a liposome containing a compound capable of modifying or modulating cell function, said liposome comprising liposome-forming phospholipids, at least a portion of which are covalently bound through their headgroups to a folate receptor binding ligand.