Table 2.

Screening of chemical library for therapeutically effective anti-staphylococcal agent.

Category	Parameter	Description
Assay	Type of assay	Inhibition of methicillin-resistant Staphylococcus aureus (MRSA) growth
	Assay strategy	Identification of growth inhibitors of Staphylococcus aureus in vitro
	Assay protocol	Broth microdilution assay according to CLSI (Clinical and Laboratory Standards Institute, 2012)
	Primary measurement	Minimum Inhibitory Concentration (MIC) against MRSA4
Library	Library size	103,873 screened
	Library composition	Synthetic chemical compounds
	Source	Drug Discovery Initiative, The University of Tokyo
Screen	Format	96-well-plates, round bottom
	Concentration(s) tested	100 μM
	Plate controls	Vancomycin (100 μg/mL) and DMSO
	Reagent/ compound dispensing system	Manual
	Detection instrument and software	Manual
	Hit criteria	Compounds inhibiting growth of S. aureus MRSA4 at a concentration of 100 μM
	Hit rate	3,383 (3.25%)
Secondary screen	Additional assay(s)	Screening for therapeutic activity using silkworm infection model infected with S. aureus
	Assay strategy	Identification of compounds that cure silkworms from S. aureus infection
	Assay protocol	According to Hamamoto et al. (2015). Fifty microliters of the compounds were injected to each silkworm.
	Concentration(s) tested	2 mM
	Hit criteria	Compounds allowing survival of 2 of 3 silkworms infected with S. aureus MSSA1
	Hit rate	3 (0.003%)
	Confirmation of hit purity and structure	HPLC, NMR, chemical synthesis, and confirmation of activity of chemically synthesized compound