RESUMEN
Transfersomes (TFs) are highly deformable hydrophilic lipid vesicles that are able to penetrate the skin barrier spontaneously because of their characteristics. Transfersomes are able to transport noninvasively low- and high-molecular-weight molecules into the body. We describe the formulation and several biologic characteristics of interleukin-2 (IL-2)- and interferon-alpha (IFNalpha )-containing TFs. TFs contain natural phosphatidylcholine and sodium cholate. Recombinant human IL-2 and human hybrid IFNalpha were added to TFs and incubated for 24 hours at 4 degrees C. Immunotransfersomes were isolated from free IL-2 and IFNalpha by filtration (Centrisart, Sartorius). The biologic activity of immunotransfersomes was measured by a cytotoxic lymphoid line assay for IL-2 and by an A549-encephalomyocarditis virus assay for IFN; concentrations of proteins were determined by the enzyme-linked immunosorbent assay (ELISA). It was possible to incorporate a large amount of IL-2 and IFN in TFs (75-80%), and the incorporated IL-2, and IFN were biologically active. The increased lipid/protein ratio (90.9/1.0) led to a growing probability of association. We were thus able to show that IL-2 and IFN are trapped by transfersomes in a biologically active form and in sufficient concentrations for immunotherapy. In upcoming experiments these IL-2- and IFN-containing TFs will be used for a transdermal approach in the murine RENCA cell line model.
Asunto(s)
Portadores de Fármacos , Interferón-alfa/administración & dosificación , Interleucina-2/administración & dosificación , Administración Cutánea , Humanos , Interferón-alfa/análisis , Interleucina-2/análisis , Proteínas Recombinantes/administración & dosificaciónRESUMEN
INTRODUCTION: Transfersomes (TF) are new highly deformable hydrophilic lipid vesicles, which are able to spontaneously penetrate the skin barrier because of their characteristics. Transfersomes are able to transport non-invasively low as well as high molecular weight molecules into the body. We describe the formulation and several biological characteristics of Interleukin-2 and Interferon-a containing TF. MATERIAL AND METHODS: TF contain natural phosphatidylcholine and sodium cholate. Recombinant human IL-2 and human hybrid interferon-alpha A/D were added to TF and incubated for 24 hours at 4 degrees C. Immunotransfersomes were isolated from free IL-2 and IFN by filtration (Centrisart, Sartorius). Biological activity of immunotransfersomes was measured by CTLL-cell-assay for IL-2 and by A549--EMCV-assay for IFN, concentrations of proteins by ELISA. RESULTS: It has been possible to incorporate a high amount of IL-2 and IFN in TF (75-80%). Incorporated IL-2 and IFN were biological active. The increase of the proportion of lipid to protein to 90.9/1 led to growing probability of association. CONCLUSION: We were able to show, that IL-2 as well as IFN is trapped by transfersomes in biological active form and in sufficient concentrations for immunotherapy. In upcoming experiments these IL-2 and IFN-containing TF are used for a transdermal approach in the murine RENCA cell line model.