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720 The extract of Deschampsia antarctica (EDA) protects fibroblasts viability from the effects of environmental oxidants and pollutants
ABSTRACTS | Pharmacology and Drug Development 720
The extract of Deschampsia antarctica (EDA) protects fibroblasts viability from the effects of environmental oxidants and pollutants A Ortiz-Espı´n1, A Delgado Rubı´n de Ce´lix2, A Brieva2, A Guerrero2, S Gonza´lez2 and F Sevilla1 1 CEBAS-CSIC, Murcia, Spain and 2 Industria Farmaceutica Cantabria, Madrid, Spain Introduction: Deschampsia Antarctica (DA) is a polyextremophile plant, able to live under high solar irradiation, salinity and oxygen concentration, low temperature and extreme dryness, thanks to special protection mechanisms against environmental aggressions. Plants cultured in a controlled environment outside their natural habitat, since the Antarctic Continent is subject to strict regulations for its protection, were used in this study. Its aim was to investigate whether an aqueous extract of the plant (EDA) preserves some of these defensive properties against common external pollutants.Materials and methods: Primary human dermal fibroblasts (HDF) were cultured in the presence or absence of EDA for 24 hours, and subsequently exposed to several environmental pollutants such as oxidants (200 mM hydrogen peroxide), tobacco smoke (5%) or heavy metals (6 mM chromium (III) and (VI), 3 mM cadmium (II) and 9 mM sodium arsenite (III)) at different concentrations to decrease cell viability but avoiding lethality. Cell viability was assessed by crystal violet. Results: Treatment of primary HDFs with hydrogen peroxide, tobacco smoke, Cr, Cd and as significantly reduced cell viability to approximately 20-50% compared to control cultures. When cultures were pretreated for 24 hours with EDA before exposition to environmental pollutants, cell viability was significantly increased. Conclusions: The aqueous EDA exerts protective effects on cell viability against the deleteri80.02ous action of several environmental pollutants common in urban areas. Further studies to deepen our knowledge on the mechanisms promoted by EDA and involved in cell defense against these agents are warranted.
722
Repurposing the clinical anti-malarial quinacrine for chaperone-mediated autophagy (CMA)-directed anti-melanoma intervention T Steinfass1, S Park1, R Justiniano1, A Hua1 and G Wondrak2 1 College of Pharmacy & UA Cancer Center, University of Arizona, Tucson, AZ and 2 University of Arizona Cancer Center, University of Arizona Dept. of Pharmacology, Tucson, AZ Malignant melanoma causes the majority of patient fatalities attributed to skin cancer, and an urgent need exists for improved therapeutic interventions. Based on cumulative evidence indicating that autophagy plays a causative role in malignant melanoma, a compound library of lysosomotropic FDA-approved anti-malarials was screened for in vitro anti-melanoma activity. The anti-malarial quinacrine (QU) was identified as a potent apoptogenic inhibitor of autophagic-lysosomal flux observable in A375 malignant melanoma cells, a finding confirmed by flow cytometry and immuno-detection of autophagy-related protein markers (LC3-I/II, SQSTM1/p62, LAMP1). Fluorescence microscopy revealed the rapid lysosomal uptake of the QU-fluorophore, antagonized by the vacuolar ATPase inhibitor bafilomycin A1. QU-induced melanoma cell death was substantiated by immuno-detection of specific apoptotis regulators (p53, p-p53-Ser15, EGR1, MCL1, PUMA, BAX), and similar cytotoxic effects were observed in BRAF-kinase inhibitor-resistant melanoma cells (A375R). Expression array analysis identified a number of QU-modulated heat shock response target genes displaying significant downregulation at the mRNA level (HSPA8, DNAJA1), an observation consistent with the previously identified role of QU as a pharmacological heat shock factor (HSF1)-inhibitor and inducer of the proteotoxic stress response. QU-induced HSPA8 downregulation, a key factor in chaperone-mediated autophagy (CMA), was confirmed by immuno-detection of lysosomal HSC70 depletion. In a subsequent murine A375 xenograft model, systemic administration of QU displayed significant anti-melanoma activity associated with CMA-inhibition and apoptogenicity in tumor tissue. Based on these QU-induced CMA-suppressive and apoptogenic pharmacodynamic effects achievable at clinically relevant dose regimens, our data suggest that this FDA-approved anti-malarial may serve as a promising drug candidate for melanoma-directed repurposing.
S124 Journal of Investigative Dermatology (2017), Volume 137
721
Efinaconazole 10% or tavaborole 5% penetrate across poly-ureaurethane, 16%: results of in vitro release testing and clinical implications of onychodystrophy in onychomycosis D Ward1, C Adigun2, T Vlahovic3, M McClellan4, K Thakker4, R Klein4 and T Elstrom1 1 Cipher Pharmaceuticals US, LLC, Charleston, SC, 2 Dermatology & Laser Center of Chapel Hill, Chapel Hill, NC, 3 Temple University School of Podiatry, Philadelphia, PA and 4 Tergus Pharma, LLC, Durham, NC Background: Poly-ureaurethane has been described for treating dry, brittle, and in general, dystrophic nails. The polymer yields a waterproof, breathable barrier to protect the nail plate. A barrier that protects but also allows a topical antifungal to permeate its shield is potentially advantageous. Protection against wetting/drying, fungal reservoirs, and microtrauma, followed by the addition of oral or topical antifungals after laboratory fungal confirmation may optimize outcomes in the treatment of onychomycosis. Methods: A vertical diffusion cell system was used to evaluate the ability of efinaconazole 10% and tavaborole 5% to penetrate across poly-ureaurethane 16%. Poly-ureaurethane 16% was applied to a 0.45mm nylon membrane and allowed to dry. Efinaconazole 10% or tavaborole 5% was then applied, and samples were pulled from the receptor chamber at various times. Reverse phase chromatography was then used to assess the penetration of each active ingredient. Results: The flux and permeability of efinaconazole were determined to be 503.9 +/- 31.9 mg/cm2/hr and 14.0 +/- 0.9 nm/sec. The flux and permeability of tavaborole were determined to be 755.5 +/290.4 mg/cm2/hr and 42.0 +/- 16.1 nm/sec. Conclusion: Flux values across the nail are reported to be 1.4 mg/cm2/day for efinaconazole and 204 mg/cm2/day for tavaborole. These values are substantially lower than the herein flux for both molecules across polyureaurethane 16%, suggesting that poly-ureaurethane 16%, if used prior to efinaconazole or tavaborole, would not limit either active ingredient to access the nail.
The extract of Deschampsia antarctica (EDA) protects fibroblasts viability from the effects of environmental oxidants and pollutants A Ortiz-Espı´n1, A Delgado Rubı´n de Ce´lix2, A Brieva2, A Guerrero2, S Gonza´lez2 and F Sevilla1 1 CEBAS-CSIC, Murcia, Spain and 2 Industria Farmaceutica Cantabria, Madrid, Spain Introduction: Deschampsia Antarctica (DA) is a polyextremophile plant, able to live under high solar irradiation, salinity and oxygen concentration, low temperature and extreme dryness, thanks to special protection mechanisms against environmental aggressions. Plants cultured in a controlled environment outside their natural habitat, since the Antarctic Continent is subject to strict regulations for its protection, were used in this study. Its aim was to investigate whether an aqueous extract of the plant (EDA) preserves some of these defensive properties against common external pollutants.Materials and methods: Primary human dermal fibroblasts (HDF) were cultured in the presence or absence of EDA for 24 hours, and subsequently exposed to several environmental pollutants such as oxidants (200 mM hydrogen peroxide), tobacco smoke (5%) or heavy metals (6 mM chromium (III) and (VI), 3 mM cadmium (II) and 9 mM sodium arsenite (III)) at different concentrations to decrease cell viability but avoiding lethality. Cell viability was assessed by crystal violet. Results: Treatment of primary HDFs with hydrogen peroxide, tobacco smoke, Cr, Cd and as significantly reduced cell viability to approximately 20-50% compared to control cultures. When cultures were pretreated for 24 hours with EDA before exposition to environmental pollutants, cell viability was significantly increased. Conclusions: The aqueous EDA exerts protective effects on cell viability against the deleteri80.02ous action of several environmental pollutants common in urban areas. Further studies to deepen our knowledge on the mechanisms promoted by EDA and involved in cell defense against these agents are warranted.
722
Repurposing the clinical anti-malarial quinacrine for chaperone-mediated autophagy (CMA)-directed anti-melanoma intervention T Steinfass1, S Park1, R Justiniano1, A Hua1 and G Wondrak2 1 College of Pharmacy & UA Cancer Center, University of Arizona, Tucson, AZ and 2 University of Arizona Cancer Center, University of Arizona Dept. of Pharmacology, Tucson, AZ Malignant melanoma causes the majority of patient fatalities attributed to skin cancer, and an urgent need exists for improved therapeutic interventions. Based on cumulative evidence indicating that autophagy plays a causative role in malignant melanoma, a compound library of lysosomotropic FDA-approved anti-malarials was screened for in vitro anti-melanoma activity. The anti-malarial quinacrine (QU) was identified as a potent apoptogenic inhibitor of autophagic-lysosomal flux observable in A375 malignant melanoma cells, a finding confirmed by flow cytometry and immuno-detection of autophagy-related protein markers (LC3-I/II, SQSTM1/p62, LAMP1). Fluorescence microscopy revealed the rapid lysosomal uptake of the QU-fluorophore, antagonized by the vacuolar ATPase inhibitor bafilomycin A1. QU-induced melanoma cell death was substantiated by immuno-detection of specific apoptotis regulators (p53, p-p53-Ser15, EGR1, MCL1, PUMA, BAX), and similar cytotoxic effects were observed in BRAF-kinase inhibitor-resistant melanoma cells (A375R). Expression array analysis identified a number of QU-modulated heat shock response target genes displaying significant downregulation at the mRNA level (HSPA8, DNAJA1), an observation consistent with the previously identified role of QU as a pharmacological heat shock factor (HSF1)-inhibitor and inducer of the proteotoxic stress response. QU-induced HSPA8 downregulation, a key factor in chaperone-mediated autophagy (CMA), was confirmed by immuno-detection of lysosomal HSC70 depletion. In a subsequent murine A375 xenograft model, systemic administration of QU displayed significant anti-melanoma activity associated with CMA-inhibition and apoptogenicity in tumor tissue. Based on these QU-induced CMA-suppressive and apoptogenic pharmacodynamic effects achievable at clinically relevant dose regimens, our data suggest that this FDA-approved anti-malarial may serve as a promising drug candidate for melanoma-directed repurposing.
S124 Journal of Investigative Dermatology (2017), Volume 137
721
Efinaconazole 10% or tavaborole 5% penetrate across poly-ureaurethane, 16%: results of in vitro release testing and clinical implications of onychodystrophy in onychomycosis D Ward1, C Adigun2, T Vlahovic3, M McClellan4, K Thakker4, R Klein4 and T Elstrom1 1 Cipher Pharmaceuticals US, LLC, Charleston, SC, 2 Dermatology & Laser Center of Chapel Hill, Chapel Hill, NC, 3 Temple University School of Podiatry, Philadelphia, PA and 4 Tergus Pharma, LLC, Durham, NC Background: Poly-ureaurethane has been described for treating dry, brittle, and in general, dystrophic nails. The polymer yields a waterproof, breathable barrier to protect the nail plate. A barrier that protects but also allows a topical antifungal to permeate its shield is potentially advantageous. Protection against wetting/drying, fungal reservoirs, and microtrauma, followed by the addition of oral or topical antifungals after laboratory fungal confirmation may optimize outcomes in the treatment of onychomycosis. Methods: A vertical diffusion cell system was used to evaluate the ability of efinaconazole 10% and tavaborole 5% to penetrate across poly-ureaurethane 16%. Poly-ureaurethane 16% was applied to a 0.45mm nylon membrane and allowed to dry. Efinaconazole 10% or tavaborole 5% was then applied, and samples were pulled from the receptor chamber at various times. Reverse phase chromatography was then used to assess the penetration of each active ingredient. Results: The flux and permeability of efinaconazole were determined to be 503.9 +/- 31.9 mg/cm2/hr and 14.0 +/- 0.9 nm/sec. The flux and permeability of tavaborole were determined to be 755.5 +/290.4 mg/cm2/hr and 42.0 +/- 16.1 nm/sec. Conclusion: Flux values across the nail are reported to be 1.4 mg/cm2/day for efinaconazole and 204 mg/cm2/day for tavaborole. These values are substantially lower than the herein flux for both molecules across polyureaurethane 16%, suggesting that poly-ureaurethane 16%, if used prior to efinaconazole or tavaborole, would not limit either active ingredient to access the nail.