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700 ITK and RLK inhibitor improves skin disease in a psoriatic mouse model
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Characteristics of patients (pts) with epidermolysis bullosa (EB) in the phase 3 ESSENCE study of SD-101 J Browning1, A Bruckner2, R Cornwall3, A Lugo-Somolinos4, H Lagast5, A Reha5, J Gault6, W Lenon6, L Reklis6, R Lazauskas7 and R Nardi6 1 Texas Dermatology & Laser Specialists, San Antonio, TX, 2 University of Colorado Denver, Aurora, CO, 3 Cincinnati Children’s Hospital, Cincinnati, OH, 4 University of North Carolina School of Medicine, Chapel Hill, NC, 5 Amicus, Cranbury, NJ, 6 Scioderm, Durham, NC and 7 Amicus Therapeutics, Inc., Cranbury, NJ EB is a group of genetic disorders defined by blistering of the skin, mucosa, and epithelial lining of organs. Currently, there are no approved treatments specific to EB. ESSENCE (NCT02384460) is an ongoing phase 3, global, multicenter, randomized, double-blind, vehicle-controlled study assessing the efficacy and safety of SD-101 in pts diagnosed with EB. While baseline characteristics for all randomized pts will be available at the time of presentation, here we report baseline characteristics of enrolled pts (n¼106) as of Dec. 2, 2016. Pts were randomized 1:1 to receive SD-101 (6.0% allantoin) cream or vehicle (0% allantoin), applied topically once daily to the entire body for 90 days. Eligible pts were aged 1 month and had a target wound (10-50 cm2) present for 21 days. Pts completing ESSENCE were eligible to enter a separate open-label study (NCT02670330) to assess the long-term efficacy and safety of SD-101. Proportions of randomized pts with EB subtype were: 11.3 % simplex; 63.2% recessive dystrophic; and 25.5% junctional non-Herlitz. Demographics were as follows: 56.6% female; 77.4% white; 5.7% African American; and 6.6% Asian. Mean (SD) age was 16.1 (15.0) yrs (range 0-67 yrs) and body mass index was 17.6 (4.4) (range 9.9-31). Randomized pts had a mean (SD) body surface area index for a total body wound burden of 9.4% (8.5%). One hundred pts (94.3%) were taking 1 concomitant medication at baseline. This study is the largest clinical trial of an investigational drug for EB. Results are expected in mid-2017.
Atopic dermatitis treatment: New insight through 3D in vitro models P Rouaud-Tinguely1, D Boudier1, C Mainzer2, V Barruche1, L Marchand1, S Bordes1 and B Closs1 1 R&D Department SILAB, Brive, France, Brive, Limousin, France and 2 SILAB Inc., Hazlet, NJ Inflammation, deterioration of the barrier function and microbiota imbalance are the three main characteristics of atopic skin. Because only dermocorticoids are used to manage skin lesions during crises, it is crucial to offer a suitable answer to the daily care of atopic skin. For this purpose, we investigated pharmacological activities of oligofructans (OF) from Ophiopogon japonicus tubers on secured in vitro 3D models mimicking atopic skin characteristics (Rouaud-Tinguely et al., 2015). Hence, tested systemically on reconstructed epidermis exposed to an inflammatory cocktail (compromised RE), OF restores the expression of 10 genes modulated in AD (>1.5 fold change). These genes are involved in the pathways of inflammation, water transport and epidermal differentiation. Topically tested on the previous model also subjected to a solution of sodium lauryl sulfate (SLS) that alters the barrier function, OF improves significantly (P < 0.05) the epidermal cohesion (synthesis of claudin-1) and differentiation (synthesis of filaggrin and loricrin), the integrity (epidermal morphology and penetration of Lucifer yellow) and the resistance (transepithelial electric resistance) of the barrier function. It also significantly (P < 0.05) restores hydration (carbonic anhydrase II expression) and down-regulates inflammation (TSLP and IL-8 synthesis). Finally, applied on RE subjected to bacterial aggression (S. aureus), OF reduces microbial adhesion and biofilm formation, thus correcting microbiota imbalance. All these results validate OF as potential therapeutic molecule also supported by in vivo and clinical assessments on young and adult atopic patients.
Chemical characterization and in vitro antioxidant activity of products from Copaifera langsdorffii and Eugenia caryophyllata M Andreassi1, G Tamasi1, C Bonechi1, A Byelyakova1, D Giustarini2, R Rossi3 and C Rossi1 1 Department of Biotechnologies Chemistry and Pharmacy, University of Siena, Siena, Toscana, Italy, 2 Department of Life Sciences, University of Siena, Italy (2), Siena, Toscana, Italy and 3 Department of Life Sciences, University of Siena, Siena, Toscana, Italy This work was aimed to investigate the chemical characterization and in vitro antioxidant activity (AA) of copaiba oleoresin and essential oil, from Copaifera langsdorffii, and clove oil from Eugenia caryophyllata. A GCMS method was validated, and applied to the above commercial products and two new formulated food supplements, having as ingredients copaiba oleoresin and docosaesaenoic acid. The quantification of b-caryophyllene (bCar) and a-humulene (aHum), was carried out via external calibration method and internal standard octhylacetate. The two copaiba products showed contents of bCar of 303.221.2 and 227.115.9 mM (35.02.5 and 30.32.1%), and aHum of 15.80.8 and 17.20.9 mM (5.50.3 and 5.20.3%). Other sesquiterpene components were found in both samples: acopaene, g-elemene and d-cadinene (3-8%). Good contents of a-himalachene and b-cubebene (6.00.4 and 9.70.6%) were found in oleoresin, respect to essential oil (1.50.1 and 1.90.1%). As regards the clove oil, the contents of bCar and aHum resulted 81.65.7 mM (13.60.9%) and 11.00.6 mM (1.50.1%), respectively, the major components being eugenol and acethyleugenol, (52.73.7 and 26.51.8% of total components). The AA was tested on HaCat cell cultures, that were incubated in PBS containing different concentrations of the five samples and irradiated with scalar UV-dose (12-90 mJ/cm2). The AA was evaluated after 24 h by MTT assay, obtaining preliminary results very promising, showing an higher protecting activity on HaCat cells from UV radiation damages, respect to control.
Dual IL-17A and IL-17F inhibition with bimekizumab provides evidence for IL-17F contribution to immune-mediated inflammatory skin response A Maroof1, T Smallie2, S Archer1, C Simpson1, M Griffiths1, D Baeten3 and S Shaw1 1 UCB, Slough, United Kingdom, 2 UCB, Slough, United Kingdom and 3 UCB, Amsterdam, Noord-Holland, Netherlands IL-17A and IL-17F are proinflammatory cytokines that share similar biologic function and structural homology1. Both are known to be upregulated in several human immune-mediated inflammatory diseases1-3, however, compared with IL-17A, the role of IL-17F is less clear. We hypothesized that IL-17F also contributes to chronic tissue inflammation within lesional skin from patients with PsA. To test this, we developed bimekizumab, a humanized monoclonal IgG1 antibody that potently and selectively neutralizes both IL-17A and IL-17F. Bimekizumab is currently in clinical development for psoriasis and PsA. Immunohistologic staining detected expression of IL-17F protein in lesional skin tissue from patients with PsA. Stimulation of normal dermal fibroblasts with recombinant IL-17F, in the presence of TNFa, induced production of proinflammatory mediators (e.g. IL-8 & IL-6), though to a lesser extent than stimulation with recombinant IL-17A. To assess the contribution of IL-17A and IL-17F in skin inflammation we developed a complex model using normal dermal fibroblasts stimulated with Th17-derived supernatant. Dual neutralization of IL-17A and IL-17F with bimekizumab demonstrated greater inhibition of IL-8 (57% lower p < 0.0001) production vs IL-17A alone. Gene transcriptional analysis also revealed lower levels of expression of 27 inflammationlinked genes including IL-6, IL-8, CXCL1 and CXCL2 with bimekizumab (dual inhibition of IL-17A & IL-17F) vs IL-17A inhibition alone, in Th17-stimulated normal dermal fibroblasts. These preclinical data indicate that dual neutralization of IL-17A and IL-17F more profoundly suppresses the inflammatory response of normal dermal fibroblasts, than IL-17A blockade alone. Dual inhibition of IL-17A and IL-17F may therefore be effective in the treatment of immune-mediated inflammatory diseases, including psoriasis and PsA.
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ITK and RLK inhibitor improves skin disease in a psoriatic mouse model JM Fuhriman1, MCG Winge1, H Haberstock-Debic2, J Oliver Funk2, M Bradshaw2 and M Marinkovich3 1 Stanford University, Stanford, CA, 2 Principia Biopharma Inc., South San Francisco, CA and 3 Stanford University School of Medicine, Stanford, CA Differentiated T lymphocytes play a key pathogenic role in the highly prevalent immunoproliferative skin disorder human psoriasis, and while a number of biologics are available which target this process, the need for improved therapies persists. PRN694, a small molecule, targets ITK and RLK, two TEC kinases activated downstream of T lymphocyte activation, which are upregulated in lesional psoriasis skin. These TEC kinases mediate signaling cascades controlling T lymphocyte proliferation, differentiation, migration, and proinflammatory cytokine production. In vitro analysis showed that PRN694 effectively inhibited IL-17A production from murine Th17 differentiated T lymphocytes. Additionally, we administered PRN694 intraperitoneally daily for 40 days in a recently described murine psoriasis model, developed in our lab (J Clin Invest 126:2661-77). PRN694 was well tolerated in treated mice and produced both marked clinical improvement as well as reduced epidermal proliferation and thickness. PRN694 also inhibited aCD3+ cell infiltration into dermal regions and reduced epidermal Stat3 activation. Inhibition of T-cell differentiation and pro-inflammatory cytokine secretion, the downstream inhibition of key epidermal psoriasis signaling pathways, the inhibition of epidermal proliferation, as well as in vivo clinical effectiveness in a well characterized mouse psoriasis model implicate PRN694 as a novel and effective psoriasis therapeutic.
S120 Journal of Investigative Dermatology (2017), Volume 137
Investigation of the absorption of allantoin in vitro skin models to support wound healing A Paller1, R Nardi2, H Do3, A Reha3, C Viereck3, H Lagast3, J Gault2, J Castelli3 and J Barth3 1 Northwestern University, Chicago, IL, 2 Scioderm, Durham, NC and 3 Amicus, Cranbury, NJ Allantoin has been investigated in wound healing in formulations with minimal or unknown dermal penetration properties. We investigated the percutaneous absorption of the novel proprietary SD-101 cream (containing 0.5%-9.0% allantoin) and vehicle in 5 in vitro models: (1) barrier-free to simulate delivery directly to the capillary bed; (2) unabraded porcine; (3) abraded porcine to simulate compromised skin; (4) intact (full thickness) human; and (5) dermis-only human to mimic loss of skin barrier function due to broken skin. Creams were tested on human cadavers and porcine donors mounted in chambers designed to maintain skin in typical in vivo conditions. Samples were collected from the solution bathing the inner surface of the skin about 2, 4, 8, 12, 24, 32, and 48h after application and analyzed using HPLC with UV and MS detection. There was evidence of allantoin absorption in all models. Absorption was lowest in intact human skin. Absorption increased with higher concentrations of allantoin in the dermis-only human model, and uptake between the barrier-free and dermis-only human model was similar. The meanSE absorption (mg) of 6% allantoin after 48h was 41.578.43 in the intact human model, 109.5239.31 in the abraded porcine model, and 1990.40167.34 in the dermis-only human model, and after 24h was 1348.30201.39 in the barrier-free model. Allantoin absorption in the dermis-only human model was slow (z8h), suggesting a long skin exposure time. Separate preclinical studies (with up to 9.0% allantoin) indicated no systemic absorption. In summary, allantoin is absorbed by the intact stratum corneum which may reduce wound formation. When damaged, as in wound models, skin absorption of allantoin increased significantly. Such findings support further investigation of SD-101 for the potential to treat wounds in a clinical setting.
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Characteristics of patients (pts) with epidermolysis bullosa (EB) in the phase 3 ESSENCE study of SD-101 J Browning1, A Bruckner2, R Cornwall3, A Lugo-Somolinos4, H Lagast5, A Reha5, J Gault6, W Lenon6, L Reklis6, R Lazauskas7 and R Nardi6 1 Texas Dermatology & Laser Specialists, San Antonio, TX, 2 University of Colorado Denver, Aurora, CO, 3 Cincinnati Children’s Hospital, Cincinnati, OH, 4 University of North Carolina School of Medicine, Chapel Hill, NC, 5 Amicus, Cranbury, NJ, 6 Scioderm, Durham, NC and 7 Amicus Therapeutics, Inc., Cranbury, NJ EB is a group of genetic disorders defined by blistering of the skin, mucosa, and epithelial lining of organs. Currently, there are no approved treatments specific to EB. ESSENCE (NCT02384460) is an ongoing phase 3, global, multicenter, randomized, double-blind, vehicle-controlled study assessing the efficacy and safety of SD-101 in pts diagnosed with EB. While baseline characteristics for all randomized pts will be available at the time of presentation, here we report baseline characteristics of enrolled pts (n¼106) as of Dec. 2, 2016. Pts were randomized 1:1 to receive SD-101 (6.0% allantoin) cream or vehicle (0% allantoin), applied topically once daily to the entire body for 90 days. Eligible pts were aged 1 month and had a target wound (10-50 cm2) present for 21 days. Pts completing ESSENCE were eligible to enter a separate open-label study (NCT02670330) to assess the long-term efficacy and safety of SD-101. Proportions of randomized pts with EB subtype were: 11.3 % simplex; 63.2% recessive dystrophic; and 25.5% junctional non-Herlitz. Demographics were as follows: 56.6% female; 77.4% white; 5.7% African American; and 6.6% Asian. Mean (SD) age was 16.1 (15.0) yrs (range 0-67 yrs) and body mass index was 17.6 (4.4) (range 9.9-31). Randomized pts had a mean (SD) body surface area index for a total body wound burden of 9.4% (8.5%). One hundred pts (94.3%) were taking 1 concomitant medication at baseline. This study is the largest clinical trial of an investigational drug for EB. Results are expected in mid-2017.
Atopic dermatitis treatment: New insight through 3D in vitro models P Rouaud-Tinguely1, D Boudier1, C Mainzer2, V Barruche1, L Marchand1, S Bordes1 and B Closs1 1 R&D Department SILAB, Brive, France, Brive, Limousin, France and 2 SILAB Inc., Hazlet, NJ Inflammation, deterioration of the barrier function and microbiota imbalance are the three main characteristics of atopic skin. Because only dermocorticoids are used to manage skin lesions during crises, it is crucial to offer a suitable answer to the daily care of atopic skin. For this purpose, we investigated pharmacological activities of oligofructans (OF) from Ophiopogon japonicus tubers on secured in vitro 3D models mimicking atopic skin characteristics (Rouaud-Tinguely et al., 2015). Hence, tested systemically on reconstructed epidermis exposed to an inflammatory cocktail (compromised RE), OF restores the expression of 10 genes modulated in AD (>1.5 fold change). These genes are involved in the pathways of inflammation, water transport and epidermal differentiation. Topically tested on the previous model also subjected to a solution of sodium lauryl sulfate (SLS) that alters the barrier function, OF improves significantly (P < 0.05) the epidermal cohesion (synthesis of claudin-1) and differentiation (synthesis of filaggrin and loricrin), the integrity (epidermal morphology and penetration of Lucifer yellow) and the resistance (transepithelial electric resistance) of the barrier function. It also significantly (P < 0.05) restores hydration (carbonic anhydrase II expression) and down-regulates inflammation (TSLP and IL-8 synthesis). Finally, applied on RE subjected to bacterial aggression (S. aureus), OF reduces microbial adhesion and biofilm formation, thus correcting microbiota imbalance. All these results validate OF as potential therapeutic molecule also supported by in vivo and clinical assessments on young and adult atopic patients.
Chemical characterization and in vitro antioxidant activity of products from Copaifera langsdorffii and Eugenia caryophyllata M Andreassi1, G Tamasi1, C Bonechi1, A Byelyakova1, D Giustarini2, R Rossi3 and C Rossi1 1 Department of Biotechnologies Chemistry and Pharmacy, University of Siena, Siena, Toscana, Italy, 2 Department of Life Sciences, University of Siena, Italy (2), Siena, Toscana, Italy and 3 Department of Life Sciences, University of Siena, Siena, Toscana, Italy This work was aimed to investigate the chemical characterization and in vitro antioxidant activity (AA) of copaiba oleoresin and essential oil, from Copaifera langsdorffii, and clove oil from Eugenia caryophyllata. A GCMS method was validated, and applied to the above commercial products and two new formulated food supplements, having as ingredients copaiba oleoresin and docosaesaenoic acid. The quantification of b-caryophyllene (bCar) and a-humulene (aHum), was carried out via external calibration method and internal standard octhylacetate. The two copaiba products showed contents of bCar of 303.221.2 and 227.115.9 mM (35.02.5 and 30.32.1%), and aHum of 15.80.8 and 17.20.9 mM (5.50.3 and 5.20.3%). Other sesquiterpene components were found in both samples: acopaene, g-elemene and d-cadinene (3-8%). Good contents of a-himalachene and b-cubebene (6.00.4 and 9.70.6%) were found in oleoresin, respect to essential oil (1.50.1 and 1.90.1%). As regards the clove oil, the contents of bCar and aHum resulted 81.65.7 mM (13.60.9%) and 11.00.6 mM (1.50.1%), respectively, the major components being eugenol and acethyleugenol, (52.73.7 and 26.51.8% of total components). The AA was tested on HaCat cell cultures, that were incubated in PBS containing different concentrations of the five samples and irradiated with scalar UV-dose (12-90 mJ/cm2). The AA was evaluated after 24 h by MTT assay, obtaining preliminary results very promising, showing an higher protecting activity on HaCat cells from UV radiation damages, respect to control.
Dual IL-17A and IL-17F inhibition with bimekizumab provides evidence for IL-17F contribution to immune-mediated inflammatory skin response A Maroof1, T Smallie2, S Archer1, C Simpson1, M Griffiths1, D Baeten3 and S Shaw1 1 UCB, Slough, United Kingdom, 2 UCB, Slough, United Kingdom and 3 UCB, Amsterdam, Noord-Holland, Netherlands IL-17A and IL-17F are proinflammatory cytokines that share similar biologic function and structural homology1. Both are known to be upregulated in several human immune-mediated inflammatory diseases1-3, however, compared with IL-17A, the role of IL-17F is less clear. We hypothesized that IL-17F also contributes to chronic tissue inflammation within lesional skin from patients with PsA. To test this, we developed bimekizumab, a humanized monoclonal IgG1 antibody that potently and selectively neutralizes both IL-17A and IL-17F. Bimekizumab is currently in clinical development for psoriasis and PsA. Immunohistologic staining detected expression of IL-17F protein in lesional skin tissue from patients with PsA. Stimulation of normal dermal fibroblasts with recombinant IL-17F, in the presence of TNFa, induced production of proinflammatory mediators (e.g. IL-8 & IL-6), though to a lesser extent than stimulation with recombinant IL-17A. To assess the contribution of IL-17A and IL-17F in skin inflammation we developed a complex model using normal dermal fibroblasts stimulated with Th17-derived supernatant. Dual neutralization of IL-17A and IL-17F with bimekizumab demonstrated greater inhibition of IL-8 (57% lower p < 0.0001) production vs IL-17A alone. Gene transcriptional analysis also revealed lower levels of expression of 27 inflammationlinked genes including IL-6, IL-8, CXCL1 and CXCL2 with bimekizumab (dual inhibition of IL-17A & IL-17F) vs IL-17A inhibition alone, in Th17-stimulated normal dermal fibroblasts. These preclinical data indicate that dual neutralization of IL-17A and IL-17F more profoundly suppresses the inflammatory response of normal dermal fibroblasts, than IL-17A blockade alone. Dual inhibition of IL-17A and IL-17F may therefore be effective in the treatment of immune-mediated inflammatory diseases, including psoriasis and PsA.
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ITK and RLK inhibitor improves skin disease in a psoriatic mouse model JM Fuhriman1, MCG Winge1, H Haberstock-Debic2, J Oliver Funk2, M Bradshaw2 and M Marinkovich3 1 Stanford University, Stanford, CA, 2 Principia Biopharma Inc., South San Francisco, CA and 3 Stanford University School of Medicine, Stanford, CA Differentiated T lymphocytes play a key pathogenic role in the highly prevalent immunoproliferative skin disorder human psoriasis, and while a number of biologics are available which target this process, the need for improved therapies persists. PRN694, a small molecule, targets ITK and RLK, two TEC kinases activated downstream of T lymphocyte activation, which are upregulated in lesional psoriasis skin. These TEC kinases mediate signaling cascades controlling T lymphocyte proliferation, differentiation, migration, and proinflammatory cytokine production. In vitro analysis showed that PRN694 effectively inhibited IL-17A production from murine Th17 differentiated T lymphocytes. Additionally, we administered PRN694 intraperitoneally daily for 40 days in a recently described murine psoriasis model, developed in our lab (J Clin Invest 126:2661-77). PRN694 was well tolerated in treated mice and produced both marked clinical improvement as well as reduced epidermal proliferation and thickness. PRN694 also inhibited aCD3+ cell infiltration into dermal regions and reduced epidermal Stat3 activation. Inhibition of T-cell differentiation and pro-inflammatory cytokine secretion, the downstream inhibition of key epidermal psoriasis signaling pathways, the inhibition of epidermal proliferation, as well as in vivo clinical effectiveness in a well characterized mouse psoriasis model implicate PRN694 as a novel and effective psoriasis therapeutic.
S120 Journal of Investigative Dermatology (2017), Volume 137
Investigation of the absorption of allantoin in vitro skin models to support wound healing A Paller1, R Nardi2, H Do3, A Reha3, C Viereck3, H Lagast3, J Gault2, J Castelli3 and J Barth3 1 Northwestern University, Chicago, IL, 2 Scioderm, Durham, NC and 3 Amicus, Cranbury, NJ Allantoin has been investigated in wound healing in formulations with minimal or unknown dermal penetration properties. We investigated the percutaneous absorption of the novel proprietary SD-101 cream (containing 0.5%-9.0% allantoin) and vehicle in 5 in vitro models: (1) barrier-free to simulate delivery directly to the capillary bed; (2) unabraded porcine; (3) abraded porcine to simulate compromised skin; (4) intact (full thickness) human; and (5) dermis-only human to mimic loss of skin barrier function due to broken skin. Creams were tested on human cadavers and porcine donors mounted in chambers designed to maintain skin in typical in vivo conditions. Samples were collected from the solution bathing the inner surface of the skin about 2, 4, 8, 12, 24, 32, and 48h after application and analyzed using HPLC with UV and MS detection. There was evidence of allantoin absorption in all models. Absorption was lowest in intact human skin. Absorption increased with higher concentrations of allantoin in the dermis-only human model, and uptake between the barrier-free and dermis-only human model was similar. The meanSE absorption (mg) of 6% allantoin after 48h was 41.578.43 in the intact human model, 109.5239.31 in the abraded porcine model, and 1990.40167.34 in the dermis-only human model, and after 24h was 1348.30201.39 in the barrier-free model. Allantoin absorption in the dermis-only human model was slow (z8h), suggesting a long skin exposure time. Separate preclinical studies (with up to 9.0% allantoin) indicated no systemic absorption. In summary, allantoin is absorbed by the intact stratum corneum which may reduce wound formation. When damaged, as in wound models, skin absorption of allantoin increased significantly. Such findings support further investigation of SD-101 for the potential to treat wounds in a clinical setting.