Sulfasalazine

Drugs for this study were purchased in solid powder form Selleck Chem, Houston, TX, United States (BPTES, Sulfasalazine, Rapamycin A769662, SnMP, P7C3, Etomoxir, BSO, GSK2194069). AGI15280, AGI-519 and GNE-140 were purchased from Adooq Bioscience. All metabolites were purchased in solid powder form from Sigma-Aldrich.
Drugs and metabolites were formulated with 8,000 g/mol molecular weight polyethylene glycol (PEG) (Polysciences, Inc.) into a 25% w/w drug-PEG-8000 composite powder and were loaded into micro-reservoirs as described in (Jonas et al., 2015 (link)). Drugs are chosen based on specific role in the immune and/or cancer metabolism.

Ferrostatin‐1 (S7243), deferoxamine (S5742), and sulfasalazine (S1576) were purchased from Selleck. β‐Mercaptoethanol (07604) was purchased from Sigma‐Aldrich. H2DCFDA (D399) and BODIPY 581/591 C11 (D3861) were purchased from Invitrogen. Dextran (M_n = 250 kDa) and chitosan (low molecular weight, deacetylated chitin) were purchased from Sigma‐Aldrich (Shanghai, P. R. China). Sodium periodate (NaIO₄, 99.5%) was obtained from Rhawn reagent (Shanghai, P. R. China). Hydroxylamine hydrochloric acid and methyl orange were purchased from Aladdin (Shanghai, P. R. China). Sodium hydroxide, diethylene glycol, and acetone were obtained from Xilong Scientific (Shantou, P. R. China). Dimethyl sulfoxide (DMSO) was obtained from Sigma‐Aldrich (Shanghai, P. R. China).

Vero E6 and Calu-3 cells were seeded using 8 × 10⁴ cells in 24-well plates. The following day, cells were treated for 3 hours before infection with the indicated doses of ademetionine (30 μM; Selleckchem), alogliptin (10 μM; Selleckchem), flucytosine (300 μM; Selleckchem), proguanil (5 nM to 500 μM; Selleckchem), sulfasalazine (5 nM to 500 μM; Selleckchem), IFN-A (1000 U/ml), dimethyl sulfoxide (DMSO; Sigma-Aldrich), or mock and infected with SARS-CoV-2 at an MOI of 0.01 in serum-free DMEM at 37°C for 24 hours before RNA or protein lysis. Infection experiments were performed under biosafety level 3 conditions.

Drugs for this study were purchased from Selleck Chemicals (Minocycline, Vorinostat (SAHA), Donepezil, 4-aminobenzoic hydrazide (ABAH), Riluzole, Paclitaxel, Sulfasalazine, SnMP, P7C3, Etomoxir, BSO, and MK2206). AGI-25696 was purchased from Adooq Bioscience (A20250). AD drugs are chosen based on differential target function for Alzheimer’s disease or specific metabolic pathway (Table 1). Drugs were formulated with polyethylene glycol (PEG) 3,400 or 8,000 (Spectrum Chemical) as described in (Jonas et al., 2015 (link)). Briefly, compounds in the ratio of 20% drug were added to 80% PEG-3450 or PEG-8000 and vortexed for 5 min above its melting point (37 °C). For insoluble drugs, a mixture of drug, PEG, and an organic solvent (ethanol or acetone) was heated to ∼37°C until completely dissolved. The solution was placed on a rotary evaporator (Buchi) for ∼30–40 min at below respective vapor pressures to completely evaporate the solvent, leaving a homogeneous solid mixture of drug and PEG.
Drug/polymer mixtures were loaded into micro-reservoirs as described in (Jonas et al., 2015 (link)). Local drug concentration is measured using drug autofluorescence or Maldi imaging mass spectrometry as described in (Jonas et al., 2015 (link); Davidson et al., 2016 (link); Jonas et al., 2016 (link)), and calibrated using homogenized drug-tissue sections of identical thickness.