Mutant Selection Windows of Azalomycin F_5a in Combination with Vitamin K₃ against Methicillin-Resistant Staphylococcus aureus

Azalomycin F5a, a 36-membered macrocyclic lactone isolated from several streptomyces strains, presented remarkable anti-methicillin-resistant Staphylococcus aureus (MRSA) activities. To improve its anti-MRSA potential and to evaluate the probability of MRSA resistant to it before development, the anti-MRSA activities of azalomycin F5a in combination with vitamin K3 were first evaluated using checkerboard assay. Then the minimal concentration inhibiting colony formation by 99% (MIC99) and mutant prevention concentration (MPC) of azalomycin F5a alone and in combination with vitamin K3 against MRSA were determined using agar plates with linear antimicrobial concentration decrease. The fractional inhibitory concentration indexes (FICIs) of 0.25 - 0.50 showed the synergistic activity of azalomycin F5a in combination with vitamin K3. The mutant selection windows (MSWs, MIC99-MPC) of azalomycin F5a alone against MRSA tested were 2.07 - 6.40 μg/mL, and the MPCs of azalomycin F5a in combination with vitamin K3 against MRSA tested were 1.60 - 3.20 μg/mL. These indicated that the MPCs of azalomycin F5a in combination could drop down to below its MIC99 alone. According to the hypothesis of MSW, the narrower MSWs of azalomycin F5a alone, even closed MSWs in combination with vitamin K3, together with their synergistic anti-MRSA activities, indicated that azalomycin F5a had a good potential to develop as a new antimicrobial agent.