Palmitoyl derivatives of interferon alpha: potential for cutaneous delivery.
Palmitoyl derivatives of interferon alpha2b (p-IFNalpha) were prepared by covalent attachment of the fatty acid to lysine residues in the protein through a reaction with N-hydroxysuccinimide palmitate ester. The p-IFNalpha was characterized by capillary electrophoresis (CE), mass spectrometry (MS), SDS-PAGE, and antiviral assay. Flow-through diffusion cells and human breast skins were used to measure cutaneous and percutaneous absorption. Formation of p-IFNalpha derivatives was demonstrated by CE to be dependent on reaction time and reagent: protein ratio. Electrospray MS of the crude p-IFNalpha mixture indicated three populations of IFNalpha derivatives with 10, 11, and 12 palmitoyl substitutions. The addition of palmitoyl residues to IFNalpha under the conditions described reduced the antiviral specific activity by 50%. However, the cutaneous absorption of p-IFNalpha was about 5-6 times greater than the parent protein. The amount of p-IFNalpha and IFN alpha in whole skin after 24 h of treatment was 2.106 +/- 1.216 microg/cm2 and 0.407 +/- 0.108 microg/cm2, respectively. Approximately two times higher flux was detected for p-IFNalpha compared to the nonfatty acylated IFNalpha. The total amount of drug diffused in 24 h was also approximately two times higher for the p-IFNalpha. The results indicate a potential for using fatty acylated derivatives of IFN alpha for dermal and transdermal delivery.
College of Pharmacy and Nutrition, and Veterinary Infectious Disease Organization, University of Saskatchewan, Saskatoon, Saskatchewan, S7N 5C9. foldvari@duke.usask.ca
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Foldvari
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Attah-Poku
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Hu
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Li
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Hughes
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Babiuk
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Kruger
D000215:Acylation; D000279:Administration, Cutaneous; D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000008:administration & dosage* / Q000737:chemistry; D004337:Drug Carriers; D019075:Electrophoresis, Capillary; D006801:Humans; D066298:In Vitro Techniques; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000008:administration & dosage* / Q000737:chemistry; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D019308:Palmitic Acid / Q000737:chemistry*; D011994:Recombinant Proteins
D000998:Antiviral Agents; D004337:Drug Carriers; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D019308:Palmitic Acid