ART26.12, A NOVEL FATTY ACID BINDING PROTEIN 5 (FABP5) INHIBITOR, SHOWS EFFICACY IN PRECLINICAL MODELS OF PSORIASIS

Description

Fatty acid-binding protein 5 (FABP5) is upregulated in psoriasis. This study assessed the efficacy of the potent, selective, and orally active FABP5 inhibitor ART26.12 in preclinical psoriasis models. ART26.12 (25 or 100 mg/kg BID) or BMS-986165 (TYK2 inhibitor; 10 mg/kg QD) were given orally for 10 days in the imiquimod mouse model. Imiquimod increased psoriasis-like symptoms. ART26.12 (25 mg/kg BID) and BMS-986165 comparably reduced psoriasis-like symptoms by Day 6 of IMQ treatment. Histopathology showed that ART26.12 reduced symptom severity, e.g., hyperkeratosis, parakeratosis, epidermal acanthosis, and inflammatory infiltrates. Proteomic analysis indicated ART26.12 rescued expression of fillagrin-2, promoted epidermal differentiation complex associated proteins and modulated PPAR, NF-kB, and PKC pathways likely downstream of lipid modulation. Lipidomic analysis showed widespread modulation including ceramides and linoleic acid derivatives. These data suggest ART26.12 may be a potential psoriasis treatment

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Proteomics analysis was done by Creative Proteomics. Skin tissue was analysed by TMT-based quantitative proteomics for healthy control (Vehicle-Vehicle), disease control (IMQ-Vehicle), IMQ-BMS-986165 (10 mg/kg) and IMQ-ART26.12 (25 mg/kg BID). Proteins were extracted from flash-frozen skin tissue by sonication and centrifugation in the presence of lysis buffer and protease inhibitors. From each sample, 100 μg of protein was transferred, reduced (10 mM TCEP, 56 °C, 1 hour), alkylated (20 mM IAA, RT dark, 1 hour), and trypsinised overnight at 37 °C. Extracted peptides were then lyophilized to near dryness and re-dissolved in 50 mM TEAB. Peptides were then labelled using TMT16plex Isobaric Label Reagent Set (Thermofisher Science), combined, and fractionated by HPLC. Peptides were identified by Nano LC-MS/MS using an Ultimate 3000 nano UHPLC system (Thermo Scientific) coupled online to a Q Exactive HF mass spectrometer (Thermo Scientific) equipped with a Nanospray Flex Ion Source (Thermo Scientific). Lipidomics analysis of imiquimod model tissue Lipidomic analysis was done by Creative Proteomics. Skin tissue was analysed by untargeted lipidomics for healthy control (Vehicle-Vehicle), disease control (IMQ-Vehicle), IMQ-BMS-986165 (10 mg/kg) and IMQ-ART26.12 (25 mg/kg BID). UPLC-MS was carried out for each sample in positive mode. To carry out lipid extraction, 50 mg samples were thawed with 1.5 ml Chloroform:MeOH (2:1, v/v), ground for 170 seconds at 65 Hz, 0.5 mL ultrapure water was added, followed by sonication for 30 minutes at 4 °C. Cwith LPC (12:0) was added as the internal standard. Samples were centrifuged (10 minutes, 3,000 rpm, 4 °C) and the lower phase transferred to a new tube dry under nitrogen. This extract was resuspended in 200 μL isopropyl alcohol:MeOH (1:1, v/v) and 5 μL LPC (12:0) was added as the internal standard. Finally, this was centrifuged (10 minutes, 12,000 rpm, 4 °C) and the supernatant transferred for LC-MS analysis. Lipids were separated and identified using UPLC combined with Q exactive MS (thermos) on an ACQUITY UPLC BEH C18 column and MS data acquired.

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