Synthetic LDL as targeted drug delivery vehicle
The present invention provides a synthetic LDL nanoparticle comprising a lipid moiety and a synthetic chimeric peptide so as to be capable of binding the LDL receptor. The synthetic LDL nanoparticle of the present invention is capable of incorporating and targeting therapeutics to cells expressing the LDL receptor for diseases associated with the expression of the LDL receptor such as central nervous system diseases. The invention further provides methods of using such synthetic LDL nanoparticles.
1. A synthetic LDL nanoparticle comprising a lipid moiety and a synthetic chimeric peptide, wherein:
the lipid moiety forms a particle of about 10-30 nm in size, and
the synthetic chimeric peptide comprises an amphipathic a-helix and an LDL receptor binding domain, wherein the LDL receptor binding domain consists of the following amino acid sequence:
(R 1 ) x -Arg-Leu-Thr-Arg-Lys-Arg-Gly-Leu-Lys-(R 2 ) y
in which
R 1 is an amino acid sequence from 1 to 40 amino acids wherein each amino acid is independently selected from the group consisting of naturally occurring amino acids and amino acid mimetics;
R 2 is an amino acid sequence from 1 to 40 amino acids wherein each amino acid is independently selected from the group consisting of naturally occuring amino acids and amino acid mimetics; and
x and y are independently selected and are equal to zero or one (SEQ ID NO:9).
2. The synthetic LDL nanoparticle of claim 1 , wherein the mean LDL nanoparticle size is about 10 nm.
3. The synthetic LDL nanoparticle of claim 1 , wherein the lipid moiety comprises a microemulsion of lipids selected from the group consisting of phospholipids, triglyceride, cholesteryl ester, and a combination thereof.
4. The synthetic LDL nanoparticle of claim 3 , wherein the microemulsion comprises phospholipids, triolein and optionally cholesteryl oleate, wherein the molar ratio among the phospholipids, triolein and cholesterol oleate is selected from the group consisting of 3:2:1, 3:1:2, 3:0.5:2 and 3:2:0.
5. The synthetic LDL nanoparticle of claim 1 , wherein the amphipathic α-helix comprises 10 to 22 amino acids.
6. The synthetic LDL nanoparticle of claim 5 , wherein the amphipathic α-helix is selected from the group consisting of Segrest peptide, 18A peptide substituted with F3 and F4, ApoAI helix 9 and ApoAI helix 10.
7. The synthetic LDL nanoparticle of claim 1 , wherein x and y are both zero (SEQ ID NO:1).
8. The synthetic LDL nanoparticle of claim 1 , wherein:
x is one;
R 1 is Asp-Trp-Leu-Lys-Ala-Phe-Tyr-Asp-Lys-Val-Ala-Glu-Lys-Leu-Lys-Glu-Ala-Phe (SEQ ID NO:2);
y is one; and
R 2 is Leu-Ala.
9. The synthetic LDL nanoparticle of claim 1 , wherein:
x is one;
R 1 is Asp-Trp-Phe-Lys-Ala-Phe-Tyr-Asp-Lys-Val-Ala-Glu-Lys-Phe-Lys-Glu-Ala-Phe (SEQ ID NO:10);
y is one; and
R 2 is Leu-Ala.
10. The synthetic LDL nanoparticle of claim 1 , wherein:
x is one;
R 1 is Tyr-Lys-Leu-Glu-Gly-Thr-Thr (SEQ ID NO:11);
y is one; and
R 2 is Leu-Ala-Thr-Ala-Leu-Ser (SEQ ID NO:12).
11. The synthetic LDL nanoparticle of claim 1 , wherein:
x is one;
R 1 is Pro-Ala-Leu-Glu-Asp-Leu-Arg-Gln-Gly-Leu-Leu-Pro (SEQ ID NO:13);
y is one; and
R 2 is Leu.
12. The synthetic LDL nanoparticle of claim 1 , wherein:
x is one;
R 1 is Asp-Trp-Leu-Lys-Ala-Phe-Tyr-Asp-Lys-Val-Ala-Glu-Lys-Leu-Lys-Glu-Ala-Phe-Pro (SEQ ID NO:14);
y is one; and
R 2 is Pro-Asp-Trp-Leu-Lys-Ala-Phe-Tyr-Asp-Lys-Val-Ala-Glu-Lys-Leu-Lys-Glu-Ala-Phe (SEQ ID NO:15).
13. The synthetic LDL nanoparticle of claim 1 , wherein:
x is one;
R 1 is Asp-Trp-Leu-Lys-Ala-Phe-Tyr-Asp-Lys-Val-Ala-Glu-Lys-Leu-Lys-Glu-Ala-Phe-Tyr-Lys-Leu-Glu-Gly-Thr-Thr (SEQ ID NO:16);
y is one; and
R 2 is Leu-Ala-Thr-Ala-Leu-Ser (SEQ ID NO:12).
14. The synthetic LDL nanoparticle of claim 1 , wherein the chimeric peptide comprises two amphipathic α-helices.
15. The synthetic LDL nanoparticle of claim 1 , wherein the synthetic LDL nanoparticle further comprieses one or more therapeutic compounds.
16. The synthetic LDL nanoparticle of claim 15 , wherein the therapeutic compound is selected from the group consisting of small organic molecules, radioisotopes, inorganic molecules, polypeptides, peptides, antibodies, siRNA, nucleic acids, bacterial toxins and a combination thereof.
17. The synthetic LDL nanoparticle of claim 15 , wherein the therapeutic compound is taken up by the lipid moiety of the synthetic LDL nanoparticle or is covalently or non-covalently attached to the amphipathic a-helix of the chimeric peptide.
18. A method for treating or preventing a central nervous system disease in a patient, the method comprising the step of administering a therapeutically effective amount of a synthetic LDL nanoparticle comprising a lipid moiety and a synthetic chimeric peptide to the patient, wherein:
the lipid moiety forms a particle of about 10-30 nm in size, and
the synthetic chimeric peptide comprises an amphipathic α-helix and an LDL receptor binding domain, wherein the LDL receptor binding domain consists of the following amino acid sequence:
(R 1 ) x -Arg-Leu-Thr-Arg-Lys-Arg-Gly-Leu-Lys-(R 2 ) y
in which
R 1 is an amino acid sequence from 1 to 40 amino acids wherein each amino acid is independently selected from the group consisting of naturally occurring amino acids and amino acid mimetics;
R 2 is an amino acid sequence from 1 to 40 amino acids wherein each amino acid is independently selected from the group consisting of naturally occuring amino acids and amino acid mimetics; and
x and y are independently selected and are equal to zero or one (SEQ ID NO:9), and wherein the synthetic LDL nanoparticle further comprises one or more therapeutic compounds,
thereby treating or preventing the disease.
19. The method of claim 18 , wherein the method of administering the synthetic LDL nanoparticle is selected from the group consisting of systemic and local.
20. The method of claim 18 , wherein the disease is cancer that occurs in the brain.
21. The method of claim 18 , wherein the disease is selected from the group consisting of GBM, astrocytoma, ependymoma, medulloblastoma, oligodendrocytoma, meningioma, pituitary adenoma, neurilemmona, metastatic carcinoma, craniopharyngioma, dermoid, epidermoid, teratoma, angiomas, vascular malformations, sarcomas, pinealoma, chordoma, and granuloma.
22. The method of claim 18 , wherein the central nervous system disease is selected from the group consisting of: stroke, epilepsy, head trauma, viral infection, bacterial infection, fungal, rickettsial, protozoan, or helminthic infections, Alzheimer's disease, Parkinson's disease, multiple sclerosis, and hereditary metabolic diseases of the brain.
23. The method of claim 18 , wherein the therapeutic compound is selected from the group consisting of small organic molecules, inorganic molecules, therapeutic peptides and proteins, antibodies, radioisotopes, siRNA and nucleic acids for gene therapy, toxins, and anti-cancer agents.
24. The method of claim 23 , wherein the anti-cancer agent is paclitaxel oleate, paclitaxel, or doxorubicin.
25. The method of claim 22 wherein the central nervous system disease is meningitis caused by picornavirus, togavirus, herpesvirus, paramyxovirus, or arenavirus; or HIV-associated cognitive dysfunction.
26. The method of claim 22 , wherein the central nervous system disease is cryptococcal meningitis, fulminant bacterial meningitis, neurotuberculosis, toxoplasmosis, or neurosyphilis.