Aprotinin polypeptides for transporting a compound across the blood-brain barrier
The invention relates to improvements in the field of drug delivery. More particularly, the invention relates to polypeptides derived from aprotinin and from aprotinin analogs as well as conjugates and pharmaceutical compositions comprising these polypeptides or conjugates. The present invention also relates to the use of these polypeptides for transporting a compound or drug across the blood-brain barrier of a mammal and in the treatment and diagnosis of neurological diseases.
1. A method for treating a mammal having a neurological disease,
the method comprising administering to the mammal a conjugate comprising:
(a) a polypeptide comprising an amino acid sequence having at least 80% identity to the sequence of Angiopep-1 (SEQ ID NO:67), wherein the polypeptide has a substitution at the position corresponding to the cysteine at position 7 of the Angiopep-1 sequence; and
(b) an antibody or a biologically active fragment thereof for treating the neurological disease, wherein the antibody or the biologically active fragment thereof is conjugated to the polypeptide, and the conjugate is capable of crossing the blood-brain barrier or an in vitro model thereof.
2. The method of claim 1 , wherein the neurological disease is a brain cancer, a brain metastasis, schizophrenia, epilepsy, Alzheimer's disease, Parkinson's disease, Huntington's disease, stroke, or obesity.
3. The method of claim 2 , wherein the brain cancer is a glioma.
4. The method of claim 3 , wherein the glioma is a glioblastoma.
5. The method of claim 1 , wherein the polypeptide comprises an amino acid sequence having at least 85% identity to the sequence of Angiopep-1 (SEQ ID NO:67).
6. The method of claim 5 , wherein the polypeptide comprises an amino acid sequence having at least 90% identity to the sequence of Angiopep-1 (SEQ ID NO:67).
7. The method of claim 1 , wherein the substitution is the substitutition of a serine at the position corresponding to the cysteine at position 7.
8. The method of claim 7 , wherein the polypeptide comprises the amino acid sequence of Angiopep-2 (SEQ ID NO:97).
9. The method of claim 7 , wherein the polypeptide consists of the amino acid sequence of Angiopep-2 (SEQ ID NO:97).
10. The method of claim 1 , wherein the antibody or the biologically active fragment thereof is a chemotherapeutic agent.
11. The method of claim 1 , wherein the administration is intra-arterial, intra-nasal, intra-peritoneal, intravenous, intramuscular, subcutaneous, transdermal, or per os.
12. The method of claim 1 , wherein the biologically active fragment of the antibody comprises an Fv fragment, an F(ab')2 fragment, or an Fab fragment.
13. The method of claim 1 , wherein the conjugate further comprises a detectable label.
14. The method of claim 13 , wherein the detectable label comprises a radioimaging agent, an isotope, a fluorescent label, a nuclear magnetic resonance label, a luminescent label, a chromophore, a chemiluminescent label, or an enzymatic label.
15. The method of claim 14 , wherein the detectable label comprises a green fluoresence protein, a histag, β-galactosidase, indium-11, technitium-99, iodine-131, a peroxidase, or a phosphatase.
16. The method of claim 1 , wherein the conjugate further comprises a linker.
17. The method of claim 16 , wherein the linker is chemical in nature.
18. The method of claim 1 , wherein the polypeptide and the antibody or the biologically active fragment thereof are joined in a fusion protein.
19. A method for transporting an antibody or a biologically active fragment thereof across the blood-brain barrier of a mammal, the method comprising administering to the mammal a conjugate comprising:
(a) a polypeptide comprising an amino acid sequence having at least 80% identity to the sequence of Angiopep-1 (SEQ ID NO:67), wherein the polypeptide has a substitution at the position corresponding to the cysteine at position 7 of the Angiopep-1 sequence; and
(b) the antibody or the biologically active fragment thereof, wherein the antibody or the biologically active fragment thereof is conjugated to the polypeptide, and the conjugate is capable of crossing the blood-brain barrier or an in vitro model thereof.
20. The method of claim 19 , wherein the polypeptide comprises an amino acid sequence having at least 85% identity to the sequence of Angiopep-1 (SEQ ID NO:67).
21. The method of claim 19 , wherein the polypeptide comprises an amino acid sequence having at least 90% identity to the sequence of Angiopep-1 (SEQ ID NO:67).
22. The method of claim 19 , wherein the substitution is the substitution of a serine at the position corresponding to the cysteine at position 7.
23. The method of claim 22 , wherein the polypeptide comprises the amino acid sequence of Angiopep-2 (SEQ ID NO:97).
24. The method of claim 23 , wherein the polypeptide consists of the amino acid sequence of Angiopep-2 (SEQ ID NO:97).
25. The method of claim 19 , wherein the conjugate further comprises a detectable label.
26. The method of claim 25 , wherein the detectable label comprises a radioimaging agent, an isotope, a fluorescent label, a nuclear magnetic resonance label, a luminescent label, a chromophore, a chemiluminescent label, or an enzymatic label.
27. The method of claim 26 , wherein the detectable label comprises a green fluoresence protein, a histag, β-galactosidase, indium-11, technitium-99, iodine-131, a peroxidase, or a phosphatase.
28. The method of claim 19 , wherein the administration is intra-arterial, intra-nasal, intra-peritoneal, intravenous, intramuscular, subcutaneous, transdermal, or per os.
29. The method of claim 19 , wherein the biologically active fragment of the antibody comprises an Fv fragment, an F(ab')2 fragment, or an Fab fragment.
30. The method of claim 19 , wherein the conjugate further comprises a linker.
31. The method of claim 30 , wherein the linker is chemical in nature.
32. The method of claim 19 , wherein the polypeptide and the antibody or the biologically active fragment thereof are joined in a fusion protein.
33. A method of treating a patient having brain cancer, the method comprising administering to the patient a conjugate comprising:
(a) a polypeptide comprising Angiopep-2 (SEQ ID NO:97); and
(b) an antibody or a biologically active fragment thereof, wherein the antibody or the biologically active fragment thereof is conjugated to the Angiopep-2.
34. The method of claim 33 , wherein administering the conjugate comprises intravenous administration.
35. The method of claim 33 , wherein the conjugate further comprises a linker.
36. The method of claim 35 , wherein the linker is chemical in nature.
37. The method of claim 33 , wherein the polypeptide and the antibody or the biologically active fragment thereof are joined in a fusion protein.