Deoxycoformycin

Mice were euthanized for collecting the BALF. Briefly, lungs were lavaged 4 times with 0.3 ml PBS containing 10 μM dipyridamole (Sigma-Aldrich), 10 μM ADA-inhibitor deoxycoformycin (MedChem Express, USA), and 10 μM adenosine 5′-(α,β-methylene) diphosphate (Sigma-Aldrich). The BALF was centrifuged (3000 g × 5 min) to remove cells and debris. Twenty microliters of BALF supernatant was analyzed by high-performance liquid chromatography (LC-10AT, Japan) using a 5 µm Hypersil ODS column (250 × 4.6 mm, Dalian Elite Analytical Instruments Co. Ltd, Dalian, China) and a gradient of solutions A (methyl alcohol) and B (0.02 M NH₄H₂PO₄, pH 5.1) at a flow rate of 1 ml/min and run. The ADO peaks were identified and quantified by known external standard curves.

BV was obtained from Takeda Pharmaceutical Company Limited (Tokyo, Japan). IgG was purchased from GeneTex (Irvine, CA, USA). Fourteen candidate drugs were assessed as potential synergistic partners of BV. Belinostat, romidepsin, and darinaparsin were purchased from Toronto Research Chemicals Inc. (North York, ON, Canada). Pralatrexate, chidamide, deoxycoformycin, dexamethasone, lenalidomide, gemcitabine, doxorubicin, etoposide, and MK2206 were purchased from MedChemExpress (Monmouth Junction, NJ, USA). Selinexor was purchased from Ark Pharm (Arlington Heights, IL, USA). Nelarabine was purchased from Sigma-Aldrich (St Louis, MO, USA).