Compositions and methods for treating B-cell malignancies
The present invention provides a targeted multi-layered drug delivery system for the delivery of cytotoxic agents to B-cells.
1. A targeted multi-layered drug delivery system comprising:
a) a first layer comprising a nanocage made by self-assembly of a plurality of Hepatitis B Virus (HBV) core protein;
b) a second layer comprising a lipid bi-layer made by self-assembly of a plurality of anionic or cationic lipids, wherein the lipid bi-layer is covalently attached to the nanocage;
c) a B-cell targeting moiety anchored into the lipid bi-layer; and
d) at least one synthetic inhibitory RNA molecule encapsulated in the nanocage.
2. The drug delivery system of claim 1 , wherein said B-cell targeting moiety is an anti-CD22 antibody, an anti-CD20 antibody, an anti-CD19 antibody, or an anti-FcR-H1 antibody.
3. The drug delivery system of claim 1 , wherein said B-cell targeting moiety is a combination of antibodies specific for two or more of the B cell surface molecules selected from the group consisting of CD22, CD20, CD19, and FcR-H1.
4. The drug delivery system of claim 1 , wherein said synthetic inhibitory RNA molecule is a dsRNA.
5. The drug delivery system of claim 4 , wherein said dsRNA is a siRNA.
6. The drug delivery system of claim 1 , wherein said synthetic inhibitory RNA molecule is an antisense RNA.
7. The drug delivery system of claim 1 , wherein said HBV core protein comprises the amino acid sequence of SEQ ID NO: 1.
8. The drug delivery system of claim 7 , wherein the glutamic acid at position 77 of SEQ ID NO: 1 is replaced with a cysteine.
9. The drug delivery system of claim 7 , wherein the aspartic acid at position 78 of SEQ ID NO: 1 is replaced with a cysteine.
10. The drug delivery system of claim 7 , wherein the alanine at position 80 of SEQ ID NO: 1 is replaced with a cysteine.
11. The drug delivery system of claim 1 , wherein the lipid bi-layer comprises cholesterol.
12. The drug delivery system of claim 1 , wherein the lipid bi-layer comprises phospholipids.
13. The drug delivery system of claim 12 , wherein the phospholipids are selected from the group consisting of phosphatidyl ethanolamine, phosphatidyl glycerol and hydrogenated soy phosphatidyl choline (HSPC).
14. The drug delivery system of claim 1 , wherein the lipid bi-layer comprises phosphatidyl glycerol, hydrogenated soy phosphatidyl choline (HSPC) and cholesterol.
15. The drug delivery system of claim 1 , wherein the lipid bi-layer is covalently attached to the nanocage through a maleimide intermediate.
16. The drug delivery system of claim 8 , wherein phosphatidyl-ethanolamine maleimide is attached to the cysteine at amino acid 77 of SEQ ID NO:1.
17. A targeted multi-layered drug delivery system comprising:
(a) a first layer comprising a nanocage comprising a plurality of modified Hepatitis B Virus (HBV) core protein;
(b) a second layer comprising a lipid bi-layer comprising a plurality of anionic or cationic lipids, wherein the lipid bi-layer is covalently attached to said first layer through a maleimide intermediate;
(c) a B-cell targeting moiety anchored to the lipid bi-layer; and
(d) at least one synthetic inhibitory RNA molecule encapsulated in the nanocage.
18. The self-assembling nanoparticle drug delivery system of claim 17 , wherein the HBV core protein comprises SEQ ID NO: 1, wherein the glutamic acid at amino acid 77 is changed to a cysteine.
19. The drug delivery system of claim 17 , wherein said B-cell targeting moiety is an anti-CD22 antibody, an anti-CD20 antibody, an anti-CD19 antibody, or an anti-FcR-H1 antibody.
20. The drug delivery system of claim 17 , wherein said synthetic inhibitory RNA molecule is a dsRNA.
21. The drug delivery system of claim 17 , wherein said dsRNA is a siRNA.
22. The drug delivery system of claim 17 , wherein said synthetic inhibitory RNA molecule is an antisense RNA.
23. A targeted multi-layered drug delivery system comprising:
(a) a first layer comprising a nanocage comprising a plurality of modified Hepatitis B Virus (HBV) core protein comprising the amino acid sequence of SEQ ID NO: 1, wherein the glutamic acid at amino acid 77 is changed to a cysteine;
(b) a second layer comprising a lipid bi-layer comprising a plurality of anionic or cationic lipids, wherein the lipid bi-layer is covalently attached to said first layer by a phosphatidyl-ethanolamine maleimide to the cysteine at amino acid 77 of SEQ ID NO: 1;
(c) a B-cell targeting moiety anchored to the lipid bi-layer; and
(d) at least one synthetic inhibitory RNA molecule encapsulated in the nanocage.
24. The drug delivery system of claim 23 , wherein said B-cell targeting moiety is an anti-CD22 antibody, an anti-CD20 antibody, an anti-CD19 antibody, or an anti-FcR-H1 antibody.
25. The drug delivery system of claim 23 , wherein said synthetic inhibitory RNA molecule is a dsRNA.
26. The drug delivery system of claim 25 , wherein said dsRNA is a siRNA.
27. The drug delivery system of claim 23 , wherein said synthetic inhibitory RNA molecule is an antisense RNA.
28. A method for delivering a drug to a B-cell comprising contacting the cell with the drug delivery system any one of claim 1 , 17 or 23 .