Patent EA-004802-B1 (EA)
Title: MODIFIED HIV PEPTIDES BASED ON gag p24 PROTEIN, ANTIGENS, VACCINE COMPOSITIONS, IMMUNOASSAY KITS AND METHOD OF DETECTING HIV-INDUCED ANTIBODIES
Inventor: SOERENSEN BIRGER
IPC: A61P31/18
Filed: 2000-03-02
Abstract: 1. Peptide derived from HIV gag p24 protein, characterized in that the peptide comprises at least nine consecutive amino acids modified compared to the native amino acid sequence SEQ ID NO: 1: Xaa1 Xaa2 Xaa3 Xaa4 Xaa5 Xaa6 Ala Xaa8 Xaa9 Gln Thr Pro Trp Xaa14 Xaa15 Xaa16 Xaa17 Xaa18 Val Xaa20 (SEQ ID NO:1) wherein: Xaa in position 1 of the peptide derivate is Lys or Arg, Xaa in position 2 is Ala, Gly, Ser or Arg, Xaa in position 3 is Leu or Met, Xaa in position 4 is Gly or Arg, Xaa in position 5 is Pro, Thr, Val, Ser, Gln or Ala, Xaa in position 6 is Gly, Ale, Lys, Arg, Gln or Glu, Xaa in position 8 is Thr or Ser, Xaa in position 9 is Leu or Ile, Xaa in position 14 is Thr, Ser or Val, Xaa in position 15 is Ala or Ser, Xaa in position 16 is Cys or Ser, Xaa in position 17 is Gln or Leu, Xaa in position 18 is Gly, Glu ar Arg, Xaa in position 20 is Gly or Arg, wherein: the terminal ends of the sequences may be free carboxyl- or amino groups, amides, acyls, acetyls or salts thereof, and two or more of the Cys residues may form part of art intrachain- of interchain disulphide binding, a -S-(CH2)p-S- or a -(CH2)p-bridge wherein p(1-8 optionally intervened by one or more heteroatoms such as O, N or S. 2. Peptide according to claim 1, characterized in that the peptide having the amino acid sequence SEQ ID NO:2 or SEQ ID NO:3. 3. Peptide derived from HIV gag p24 protein, characterized in that the peptide comprises at least six consecutive amino acids modified compared to the native amino acid sequence SEQ ID NO: 4: Xaa1 Xaa2 Xaa3 Xaa4 Xaa5 Gly Leu Asn Pro Leu Val [Gly]n Xaa12 Xaa13 Tyr Xaa15 Pro Xaa17 Xaa18 Ile Leu Xaa21 Xaa22 (SEQ ID NO:4) wherein: Xaa in position 1 is Arg, Lys, Asp or none, Xaa in position 2 is Trp, Gly, Lys or Arg, Xaa in position 3 is Ile, Leu, Val or Met, Xaa in position 4 is Ile, Val or Leu, Xaa in position 5 is Leu, Met, Val or Pro, Xaa in position 12 is Arg, Lys, Xaa in position 13 is Met or Leu, Xaa in position 15 is Ser, Cys or Gln, Xaa in position 17 is Thr, Val, Ile, Ser or Ala, Xaa in position 18 is Ser, Gly ar Thr, Xaa in position 21 is Asp, Glu, Cys or Gly, Xaa in position 22 is Gly or none, and n (0, 1 or 3; wherein: the terminal ends of the sequences may be free carboxyl- or amino groups, amides, acyls, acetyls or salts thereof, and two or more of the Cys residues may form part of art intrachain- of interchain disulphide binding, a -S-(CH2)p-S- or a -(CH2)p-bridge wherein p(1-8 optionally intervened by one or more heteroatoms such as O, N or S. 4. Peptide according to claim 3, characterized in that the peptide having the amino acid sequence SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7 and SEQ ID NO:8. 5. Peptide derived from HIV gag p24 protein, characterized in that the peptide comprises at least six consecutive amino acids modified compared to the native amino acid sequence SEQ ID NO: 9: Xaa1 Xaa2 Xaa3 Pro Ile Pro Xaa7 Xaa8 Xaa9 Xaa10 Xaa11 Xaa12 [Gly]n Xaa13 Xaa14 Xaa15 Xaa16 Xaa17 Xaa18 Xaa19 Xaa20 Xaa21 Xaa22 Xaa23 Xaa24 (SEQ ID NO:9) wherein: Xaa in position 1 is Asn, Ser, Gly, His, Ala, Pro, Arg or none, Xaa in position 2 is Asn, Ala or Lys, Xaa in position 3 is Pro, Gln, Gly, Ile or Leu, Xaa in position 7 is Val or Ala, Xaa in position 8 is Gly or Lys, Xaa in position 9 is Glu, Asp, Lys, Phe or Thr, Xaa in position 10 is Ile, Met, Val or Leu, Xaa in position 11 is Tyr, Leu or none, Xaa in position 12 is Ser or none, Xaa in position 13 is Arg or none, Xaa in position 14 is Asp, Arg, Trp, Ala or none, Xaa in position 15 is Ile or none, Xaa in position 16 is Tyr or none, Xaa in position 17 is Lys or Arg, Xaa in position 18 is Arg, Lys or Asp, Xaa in position 19 is Trp or Gly, Xaa in position 20 is Ile, Met, Val, Gln or Ala, Xaa in position 21 is Ile, Val or Ala, Xaa in position 22 is Leu, Met or Val, Xaa in position 23 is Gly or Cys Xaa in position 24 is Leu or none, and n (0, 1 or 3; wherein: the terminal ends of the sequences may be free carboxyl- or amino groups, amides, acyls, acetyls or salts thereof, and two or more of the Cys residues may form part of art intrachain- of interchain disulphide binding, a -S-(CH2)p-S- or a -(CH2)p-bridge wherein p(1-8 optionally intervened by one or more heteroatoms such as O, N or S. 6. Peptide according to claim 1, characterized in that that the peptide having the amino acid sequence SEQ ID NO 10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13 or SEQ ID NO:14. 7. Peptide derived from HIV gag p24 protein, characterized in that the peptide comprises at least six consecutive amino acids modified compared to the native amino acid sequence SEQ ID NO: 15: Xaa1 Xaa2 Ile Ile Xaa5 Xaa6 Xaa7 Xaa8 Xaa9 Leu Xaa11 [Gly]n, [Arg]m Xaa12 Xaa13 Xaa14 Xaa15 Xaa16 Xaa17 Xaa18 Xaa19 Xaa20 Xaa21 Xaa22 Xaa23 Xaa24 Xaa25 (SEQ ID NO:15) wherein: Xaa in position 1 is Pro, Lys, Arg or none, Xaa in position 2 is Glu, Arg, Phe or Lys, Xaa in position 5 is Pro or Thr, Xaa in position 6 is Met, Thr or Nleu, Xaa in position 7 is Phe or Leu, Xaa in position 8 is Ser, Thr, Ala or Met, Xaa in position 9 is Ala, Glu or Leu, Xaa in position 11 is Ser or none, Xaa in position 12 is Ala, Arg or none, Xaa in position 13 is Ile, Leu or none, Xaa in position 14 is Ser, Ale, Leu or none, Xaa in position 15 is Tyr, Glu ar Asp, Xaa in position 16 is Gly or Asp, Xaa in position 17 is Ala or Leu, Xaa in position 18 is Thr, Ile, Val, Leu or Asn, Xaa in position 19 is Pro, Thr or Ser, Xaa in position 20 is Tyr, Phe, Nleu, His or Gln, Xaa in position 21 is Asp, Asn, Leu or Ala, Xaa in position 22 is Leu, Ile, Val or Asn, Xaa in position 23 is Asn, Tyr, Cys or Gly, Xaa in position 24 is Thr, Met, Ile, Ale, Val or none, Xaa in position 25 is Gly or none and n =1, 2 or 3 and m= 0, 1, 2 or 3 independent of each other wherein: the terminal ends of the sequences may be free carboxyl- or amino groups, amides, acyls, acetyls or salts thereof, and two or more of the Cys residues may form part of art intrachain- of interchain disulphide binding, a -S-(CH2)p-S- or a -(CH2)p-bridge wherein p(1-8 optionally intervened by one or more heteroatoms such as O, N or S. 8. Peptide according to claim 7, characterized in that that the peptide having the amino acid sequence SEQ ID NO 16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19 or SEQ ID NO:20. 9. HIV-antigen, characterized in that it comprises at least one peptide according to any one of claims 1-8. 10. Antigen according to claim 9 comprising at least one peptide of claim 1, at least one peptide of claim 3, at least one peptide of claim 5 and at least one peptide of claim 7. 11. Vaccine composition, characterized in that it comprises an antigen according to claim 9 with a pharmaceutically acceptable diluent and optionally an adjuvant, carrier and/or vehicle and optionally additional immunostimulatory compound(s). 12. Vaccine composition according to claim 11, characterized in that comprises at least four peptides, wherein at least one is selected from at least one peptide of claim 1, at least one peptide of claim 3, at least one peptide of claim 5 and at least one peptide of claim 7. 13. Vaccine composition according to claim 8, characterized in that it comprises the peptides of the SEQ ID NO:3 of claim 2, SEQ ID NO:6 of claim 4, SEQ ID NO:11 of claim 6 and SEQ ID NO:18 of claim 8. 14. Vaccine composition according to any one of claims 12-13, characterized in that the peptides are dissolved in a saline water solution and the optional immunostimulatory compound is a granulocyte macrophage growth factor. 15. Vaccine composition according to any one of claims 12-14, characterized in that the composition comprises an adjuvant selected from the group Monophosphoryl Lipid A (MPL(RTM)), Freund's complete or incomplete adjuvant or aluminum hydroxyl. 16. A method of detecting antibodies, induced by a HIV or HIV-specific peptides or proteins, in a sample of body fluid, characterized in that subjecting the said sample to an immunoassay, wherein the antigen(s) its/are selected from the peptides according to any one of claims 9-10. 17. An immunoassay kit for the detection of antibodies, induced by a HIV or HIV specific peptides or proteins, in a sample of body fluid, characterized in that the diagnostic antigen is an antigen according to any one of claims 9-10. 18. Antibody, characterized in that it is capable of selectively reacting with the antigen of claims 9 and 10.