Methods of promoting uptake and nuclear accumulation of polyamides in eukaryotic cells

• US 20030109448A1
• Filed: 11/07/2002
• Published: 06/12/2003
• Est. Priority Date: 11/07/2001
• Status: Abandoned Application

• Current Assignee: Pharmacia Corporation (Pfizer Inc.)
• Original Assignee: Pharmacia Corporation (Pfizer Inc.)
• Inventors: Crowley, Kathleen S., Schweitzer, Barbara A., Phillion, Dennis P., Bashkin, James K.
• Application Number:
  US10/289,677
• Publication Number:
  US 20030109448A1
• Time in Patent Office: Days
• US Class Current: 514/12
• CPC Class Codes: A61K 31/275 Nitriles; Isonitriles
• CPC Class Codes: A61K 31/40 having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
• CPC Class Codes: A61K 31/495 having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazinesA61K31/48 takes precedence ; with three nitrogen atoms A61K31/53
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/63 Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression

Abstract

A method for enhancing cellular uptake and redistributing gene regulating polyamides from the extranuclear areas in eukaryotic cells to the nucleus thereof. The method consists of administering to the eukaryotic cells a molecular trafficking compound. The administered molecular trafficking compound affects cellular pathways and mechanisms that accumulate polyamides in cytoplasmic vessicles and/or efflux polyamides from the intracellular regions of cell. The method also includes modifying polyamides to contain negatively-charged or acidic moieties wherein the moieties inhibit the accumulation of polyamides in cellular lysosomes. By affecting the cellular pathways and mechanisms, the polyamides are redistributed throughout the cell, whereby nuclear accumulation of the polyamides is enhanced.

Claims

1. A method for modulating the distribution of a polyamide within eukaryotic cells, the method comprising:

administering to the eukaryotic cells a molecular trafficking compound selected from the group consisting of P-glycoprotein inhibitors, ATPase affecting chemicals, pH or proton gradient disrupters, calcium channel blockers, ATP depleting chemicals, sodium/potassium channel blockers, MRP inhibitors, protein kinase inhibitors, Multidrug Resistance Compounds and combinations thereof, and

administering the polyamide to the eukaryotic cells.

2. The method of claim 1, wherein the eukaryotic cell is a mammalian cell.

3. A method of claim 1, wherein the polyamide is distributed from the extranuclear region of the cell to the nucleus of the cell.

4. The method of claim 1, further comprising administering, simultaneously or sequentially, the molecular trafficking compound and the polyamide.

5. The method of claim 4, wherein the molecular trafficking compound and the polyamide are simultaneously administered as a mixture.

6. The method of claim 1, wherein more than one polyamide and one molecular trafficking compound are administered.

7. The method of claim 1, wherein one polyamide and more than one molecular trafficking compound are administered.

8. The method of claim 1, wherein more than one polyamide and more than one molecular trafficking compound are administered.

9. The method of claim 1, wherein the molecular trafficking compound is a P-glycoprotein inhibitor.

10. The method of claim 1, wherein the molecular trafficking compound is an ATPase affecting chemical.

11. The method of claim 1, wherein the molecular trafficking compound is a pH or proton gradient disrupter.

12. The method of claim 1, wherein the molecular trafficking compound is a calcium channel blocker.

13. The method of claim 1, wherein the molecular trafficking compound is an ATP depleting chemical.

14. The method of claim 1, wherein the molecular trafficking compound is a sodium/potassium channel blocker.

15. The method of claim 1, wherein the molecular trafficking compound is a MRP inhibitor.

16. The method of claim 1, wherein the molecular trafficking compound is a protein kinase inhibitor.

17. The method of claim 1, wherein the molecular trafficking compound is a Multidrug Resistance Compound.

18. A method for modulating the expression of a gene in cells of a eukaryotic organism comprising:

administering a molecular trafficking compound selected from the group consisting of P-glycoprotein inhibitors, ATPase affecting chemicals, pH or proton gradient disrupters, calcium channel blockers, ATP depleting chemicals, sodium/potassium channel blockers, MRP inhibitors, protein kinase inhibitors, Multidrug Resistance Compounds and combinations thereof, and

administering one or more polyamides to the eukaryotic cells.

19. The method of claim 18, wherein the eukaryotic organism is a mammal.

20. The method of claim 18, wherein the polyamide is a polyamide that contains an acidic moiety.

21. A method for modulating the distribution of a polyamide within eukaryotic cells, the method comprising, conjugating an acidic moiety to the polyamide to produce a modified polyamide and administering the modified polyamide to eukaryotic cells.

23. The method of claim 21 wherein the the acidic moiety is conjugated to an amine present on the polyamide.

24. The method of claim 23, wherein the amine is located on an amine tail of the polyamide.

25. The method of claim 24, wherein the amine is a primary amine.

26. The method of claim 24 wherein the amine tail of the polyamide comprises N-methylamino, di-propylamine.

27. The method of claim 21 wherein the acidic moiety is conjugated to the polyamide with a linking group wherein the linking group is selected from the group consisting of acryl, aromatic, alkyl, allyl, and polyester groups.

22. The method of 21 wherein the acidic moiety is selected from the group consisting of fluorescien, phenol, carboxylic acid, HSO₃, and H_nPO₄ wherein n=1 to 3.

28. A composition for modulating the expression of a gene in a eukaryotic cell, the composition comprising a polyamide and a molecular trafficking compound wherein the molecular trafficking compound is selected from the group consisting of P-glycoprotein inhibitors, ATPase affecting chemicals, pH or proton gradient disrupters, calcium channel blockers, ATP depleting chemicals, sodium/potassium channel blockers, MRP inhibitors, protein kinase inhibitors, Multidrug Resistance Compounds and combinations thereof.

29. The composition of claim 28, wherein the composition comprises more than one polyamide.

30. The composition of claim 28, wherein the polyamide is a polyamide that contains an acidic moiety.