Concanavalin a (cona)

Whole lace plant cultures were aseptically propagated as described in Gunawardena et al. (2020) [48 (link)]. Newly obtained corms of lace plants [A. madagascariensis (Mirbel) H. Bruggen] were obtained from The PlantGuy (Alberta, Canada) and cultured in GA-7 Magenta boxes, embedded fully in solid MS media [100 ml of 1.5% plant tissue culture agar (w/v, Phytotech Laboratories) in liquid MS (3% sucrose (w/v), 0.01% Myo-inositol (w/v), 0.215% MS basal salts (w/v) Phytotech Laboratories), 0.0025% thiamine-HCl (v/v), pH 5.7] and then submerged in 150 ml of liquid MS. Whole plant cultures were grown at 24°C and exposed to 12 h light: 12 h dark cycles with levels of 125 μmol m^-2 s^-1 daylight deluxe fluorescent light bulbs (Philips). Cultured plants were only selected for pharmacological experimentation after 30 d of growth and the production of 3 perforated mature leaves to control for variation in plant growth.
Plants selected for pharmacological experiments were treated for 1 week with either (i) 5 μM rapamycin (Enzo Scientific, BML-275), (ii) 5 μM wortmannin (Cayman Chemical, 10010591), or (iii) 1 μM ConA (Santa Cruz Biotechnology, sc-202111). Control plants received an equal volume of dimethyl sulfoxide DMSO (<0.1% v/v; BioShop Canada, DMS666). A minimum of six replicates were performed for each group (four groups total).

Pentoxifylline (PTX), dimethylsulfoxide (DMSO), polyethylene glycol (PEG) 400, LPS (E. coli 055:B5 serotype), incomplete Freund’s adjuvant (IFA), and RPMI 1640 medium were purchased from Sigma-Aldrich (Germany). ConA was purchased from Santa Cruz Biotechnology (USA). Porcine collagen type II (2 mg mL⁻¹ in 0.05 M acetic acid) was obtained from Chondrex, Inc. (USA). Vinpocetine, EHNA, milrinone, zaprinast, and papaverine were purchased from Cayman Chemical (USA). (±)-LSF was obtained from the Department of Organic Chemistry, Faculty of Chemistry, Jagiellonian University (Krakow, Poland). GRMS-55 and 4-(8-((Furan-2-ylmethyl)amino)-1,3-dimethyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-7-yl)-N′-(2-hydroxybenzylidene)butanehydrazide were obtained from the Department of Medicinal Chemistry and rolipram from the Department of Organic Chemistry, Faculty of Pharmacy, Jagiellonian University Medical College (Krakow, Poland). Temazepam was a gift from Polfa (Poland). Other reagents and solvents were of HPLC or analytical reagent grade and were purchased from Merck (Germany).

Compound 34 was designed and synthetized in the Department of Medicinal Chemistry, Faculty of Pharmacy, Jagiellonian University Medical College, Kraków, Poland [25 (link)]. ConA was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). PEG400 and DMSO were obtained from Sigma-Aldrich (Steinheim, Germany). All other reagents, chemicals, and solvents were of HPLC or analytical grade and were purchased from Merck (Darmstadt, Germany).