Preparation of antibody or an antibody fragment-targeted immunoliposomes for systemic administration of therapeutic or diagnostic agents and uses thereof

1. A method of preparing an antibody- or antibody fragment-targeted cationic immunoliposome complex comprising:

(a) preparing an antibody or antibody fragment;

(b) mixing said antibody or antibody fragment with a cationic liposome to form a cationic immunoliposome, wherein said antibody or antibody fragment is complexed with said cationic liposome, but is not chemically conjugated to said cationic liposome, and wherein said antibody or antibody fragment does not comprise a lipid tag; and

(c) mixing said cationic immunoliposome with a small molecule inhibitor to form said antibody- or antibody fragment-targeted-cationic immunoliposome complex, wherein said antibody- or antibody fragment-targeted-cationic immunoliposome is about 50-500 nm in size.

2. The method of claim 1 , wherein an antibody is mixed with said cationic liposome.

3. The method of claim 1 , wherein an antibody fragment is mixed with said cationic liposome.

4. The method of claim 3 , wherein said antibody fragment is a single chain Fv fragment.

5. The method of claim 4 , wherein said antibody fragment is an anti-transferrin receptor single chain Fv (TfRscFv).

6. The method of claim 1 , wherein said antibody or antibody fragment is an anti-HER-2 antibody or antibody fragment.

7. The method of claim 1 , wherein said antibody fragment comprises a cysteine moiety at a carboxy terminus prior to being mixed with said cationic liposome.

8. The method of claim 1 , wherein said cationic liposome comprises a mixture of one or more cationic lipids and one or more neutral or helper lipids.

9. The method of claim 1 , wherein said antibody or antibody fragment is mixed with said cationic liposome at a ratio in the range of about 1:20 to about 1:40 (w:v).

10. The method of claim 1 , wherein said cationic liposome comprises a mixture of dioleoyltrimethylammonium phosphate with dioleoylphosphatidylethanolamine and cholesterol, a mixture of dioleoyltrimethylammonium phosphate with cholesterol, a mixture of dimethyldioctadecylammonium bromide with dioleolphosphatidylethanolamine and cholesterol, a mixture of dimethyldioctadecylammonium bromide with dioleoylphosphatidylethanolamine, a mixture of dimethyldioctadecylammonium bromide with cholesterol, a mixture of dioleiyltrimethylammonium phosphate with dioleoylphosphatidylethanolamine.

11. The method of claim 1 , wherein said cationic immunoliposome is mixed with said small molecule inhibitor at a molar ratio in the range of about 0.2:7 to about 14:7 (small molecule inhibitor:immunoliposome).

12. The method of claim 1 , wherein said cationic immunoliposome is mixed with said small molecule inhibitor at a molar ratio of about 7:7 (small molecule inhibitor:immunoliposome).

13. The method of claim 1 , wherein said small molecule inhibitor has a molecular weight of less than about 5000 Daltons.

14. The method of claim 13 , wherein said small molecule inhibitor has a molecular weight of less than about 1000 Daltons.

15. The method of claim 14 , wherein said small molecule inhibitor has a molecular weight of about 300 to about 700 Daltons.

16. The method of claim 1 , wherein said small molecule inhibitor has at least one pKa in the range of about 2 to about 9.

17. The method of claim 1 , wherein said small molecule inhibitor is an anticancer small molecule inhibitor.

18. The method of claim 1 , wherein said small molecule inhibitor is selected from the group consisting of GMC-5-193, YK-3-250, imatinib mesylate, erlotinib hydrochloride, sunitinib malate, gefitinib, and analogs and derivatives thereof.

19. A cationic immunoliposome complex prepared by the method of claim 1 .

20. An antibody- or antibody fragment-targeted cationic immunoliposome complex comprising a cationic liposome, an antibody or antibody fragment, and a small molecule inhibitor, wherein said antibody or antibody fragment is complexed with said cationic liposome, but is not chemically conjugated to said cationic liposome, wherein said antibody or antibody fragment does not comprise a lipid tag, and wherein said antibody- or antibody fragment-targeted cationic immunoliposome is about 50-500 nm in size.

21. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor is encapsulated within said cationic liposome.

22. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor is contained within a hydrocarbon chain region of said cationic liposome.

23. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor is associated with an inner or outer monolayer of said cationic liposome.

24. The cationic immunoliposome complex of claim 20 , wherein said antibody fragment is a single chain Fv fragment.

25. The cationic immunoliposome complex of claim 20 , wherein said antibody fragment is an anti-transferrin receptor single chain Fv (TfRscFv).

26. The cationic immunoliposome complex of claim 20 , wherein said antibody or antibody fragment is an anti-HER-2 antibody or antibody fragment.

27. The cationic immunoliposome complex of claim 20 , wherein said cationic liposome comprises a mixture of one or more cationic lipids and one or more neutral or helper lipids.

28. The cationic immunoliposome complex of claim 20 , wherein said antibody or antibody fragment and said cationic liposome are present at a ratio in the range of about 1:20 to about 1:40 (w:w).

29. The cationic immunoliposome complex of claim 20 , wherein said cationic liposome comprises a mixture of dioleoyltrimethylammonium phosphate with dioleoylphosphatidylethanolamine and cholesterol, a mixture of dioleoyltrimethylammonium phosphate with cholesterol, a mixture of dimethyldioctadecylammonium bromide with dioleoylphosphatidylethanolamine and cholesterol, a mixture of dimethyldioctadecylammonium bromide with dioleoylphosphatidylethanolamine, a mixture of dimethyldioctadecylammonium bromide with cholesterol, or a mixture of dioleoyltrimethylammonium phosphate with dioleoylphosphatidylethanolamine.

30. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor and said cationic immunoliposome are present at a molar ratio in the range of about 0.2:7 to about 14:7 (small molecule inhibitor:immunoliposome).

31. The cationic immunoliposome complex of claim 20 , wherein small molecule inhibitor and said cationic immunoliposome are present at a molar ratio of about 7:7 (small molecule inhibitor:immunoliposome).

32. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor has a molecular weight of less than about 5000 Daltons.

33. The cationic immunoliposome complex of claim 32 , wherein said small molecule inhibitor has a molecular weight of less than about 1000 Daltons.

34. The cationic immunoliposome complex of claim 33 , wherein said small molecule inhibitor has a molecular weight of about 300 to about 700 Daltons.

35. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor has al least one pKa the range of about 2 to about 9.

36. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor is an anticancer small molecule inhibitor.

37. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor is selected from the group consisting of GMC-5-193, YK-3-250, imatinib mesylate, erlotinib hydrochloride, sunitinib malate, gefitinib, and analogs and derivatives thereof.

38. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor and said cationic immunoliposome are present at a molar ratio in the range of about 2:7 to about 14:7 (small molecule inhibitor:immunoliposome).

39. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor and said cationic immunoliposome are present at a molar ratio in the range of about 4:7 to about 10:7 (small molecule inhibitor:immunoliposome).

40. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor and said cationic immunoliposome are present at a molar ratio in the range of about 5:7 to about 8:7 (small molecule inhibitor:immunoliposome).

41. The cationic immunoliposome complex of claim 20 , wherein said small molecule inhibitor and said cationic immunoliposome are present at a molar ratio in the range of about 2.8:7 (small molecule inhibitor:immunoliposome).

42. The cationic immunoliposome complex of claim 30 , wherein said small molecule inhibitor is selected from the group consisting of GMC-5-193, YK-3-250, imatinib mesylate, erlotinib hydrochloride, sunitinib malate, gefitinib, and analogs and derivatives thereof.

43. The cationic immunoliposome complex of claim 31 , wherein said small molecule inhibitor is selected from the group consisting of GMC-5-193, YK-3-250, imatinib mesylate, erlotinib hydochloride, sunitinib malate, gefitinib, and analogs and derivatives thereof.

44. The cationic immunoliposome complex of claim 38 , wherein said small molecule inhibitor is selected from the group consisting of GMC-5-193, YK-3-250, imatinib mesylate, erlotinib hydochloride, sunitinib malate, gefitinib, and analogs and derivatives thereof.

45. The cationic immunoliposome complex of claim 39 , wherein said small molecule inhibitor is selected from the group consisting of GMC-5-193, YK-3-250, imatinib mesylate, erlotinib hydochloride, sunitinib malate, gefitinib, and analogs and derivatives thereof.

46. The cationic immunoliposome complex of claim 40 , wherein said small molecule inhibitor is selected from the group consisting of GMC-5-193, YK-3-250, imatinib mesylate, erlotinib hydochloride, sunitinib malate, gefitinib, and analogs and derivatives thereof.

47. The cationic immunoliposome complex of claim 41 , wherein said small molecule inhibitor is selected from the group consisting of GMC-5-193, YK-3-250, imatinib mesylate, erlotinib hydochloride, sunitinib malate, gefitinib, and analogs and derivatives thereof.