title: Synthetic stigmastanes with dual antiherpetic and immunomodulating activities inhibit ERK and Akt signaling pathways without binding to glucocorticoid receptors.
abstract: BACKGROUND
  We have previously shown that some synthetic hydroxylated stigmastanes derived from plant sterols inhibit in vitro HSV-1 replication in ocular cell lines and decrease cytokine production in stimulated macrophages, suggesting that these steroids might combine antiviral and immunomodulating properties. In this paper we report the synthesis of some analogs fluorinated at C-6 in order to study the effect of this modification on bioactivity.
  
  
  METHODS
  The following methods were used: organic synthesis of fluorinated analogs, cytotoxicity determination with MTT assays, cytokine production quantification with ELISAs, glucocorticoid activity determination by displacement assays, immunofluorescence and transcriptional activity assays, studies of the activation of signaling pathways by Western blot, antiviral activity evaluation through virus yield reduction assays.
  
  
  RESULTS
  We report the chemical synthesis of new fluorinated stigmastanes and show that this family of steroidal compounds exerts its immunomodulating activity by inhibiting ERK and Akt signaling pathways, but do not act as glucocorticoids. We also demonstrate that fluorination enhances the antiviral activity.
  
  
  CONCLUSIONS
  Fluorination on C-6 did not enhance the anti-inflammatory effect, however, an increase in the in vitro antiviral activity was observed. Thus, our results suggest that it is possible to introduce chemical modifications on the parent steroids in order to selectively modulate one of the effects.
  
  
  CONCLUSIONS
  This family of steroids could allow the development of an alternative treatment for ocular immunopathologies triggered by HSV-1, without the undesirable side effects of the currently used drugs.
authors.affiliation: Laboratorio de Virología: agentes antivirales y citoprotectores, Departamento de Química Biológica - IQUIBICEN, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Buenos Aires C1428EGA, Argentina.
authors.forename: Flavia M
authors.initials: FM
authors.lastname: Michelini
authors.affiliation: Laboratorio de Virología: agentes antivirales y citoprotectores, Departamento de Química Biológica - IQUIBICEN, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Buenos Aires C1428EGA, Argentina.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Bueno
authors.affiliation: IBYME (CONICET) and Departamento de Química Biológica - IQUIBICEN, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Buenos Aires C1428EGA, Argentina.
authors.forename: Alejandro M
authors.initials: AM
authors.lastname: Molinari
authors.affiliation: IBYME (CONICET) and Departamento de Química Biológica - IQUIBICEN, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Buenos Aires C1428EGA, Argentina.
authors.forename: Mario D
authors.initials: MD
authors.lastname: Galigniana
authors.affiliation: Departamento de Química Orgánica and UMYMFOR (CONICET - Facultad de Ciencias Exactas y Naturales), Universidad de Buenos Aires, Ciudad Universitaria, Buenos Aires C1428EGA, Argentina.
authors.forename: Lydia R
authors.initials: LR
authors.lastname: Galagovsky
authors.affiliation: Laboratorio de Virología: agentes antivirales y citoprotectores, Departamento de Química Biológica - IQUIBICEN, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Buenos Aires C1428EGA, Argentina.
authors.forename: Laura E
authors.initials: LE
authors.lastname: Alché
authors.affiliation: Departamento de Química Orgánica and UMYMFOR (CONICET - Facultad de Ciencias Exactas y Naturales), Universidad de Buenos Aires, Ciudad Universitaria, Buenos Aires C1428EGA, Argentina. Electronic address: jar@qo.fcen.uba.ar.
authors.forename: Javier A
authors.initials: JA
authors.lastname: Ramírez
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002522:Chlorocebus aethiops; D016207:Cytokines / Q000096:biosynthesis; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors*; D057809:HEK293 Cells; D018259:Herpesvirus 1, Human / Q000187:drug effects*; D006801:Humans; D007155:Immunologic Factors / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects / Q000276:immunology; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors* / Q000502:physiology; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D012855:Sitosterols / Q000494:pharmacology*; D013265:Stigmasterol / Q000494:pharmacology*; D014709:Vero Cells
chemical_list: D000998:Antiviral Agents; D016207:Cytokines; D007155:Immunologic Factors; D011965:Receptors, Glucocorticoid; D012855:Sitosterols; D013265:Stigmasterol; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: Antiherpetic; Cytokines; Fluorinated steroids; Signaling pathways; Synthetic steroids