Finafloxacin was supplied by MerLion Pharmaceuticals Pte Ltd. For the in vitro assays, 100 mg of Finafloxacin or ciprofloxacin (Sigma Aldrich Ltd., United Kingdom) was added to 9 ml of sterile water and 1 ml of 1 M sodium hydroxide to make a working stock of 10 mg/ml. The equivalent concentration of sodium hydroxide, used to prepare the antibiotics, was included as a control for growth of bacteria. This control was included in all in vitro assays.
For the in vivo studies, a 15 mg/ml solution of Finafloxacin was prepared by adding 2.1 ml of 0.01 M Tris buffer to 44 mg of Finafloxacin (containing 37.5 mg of active ingredient). 200 μl of 1 M sodium hydroxide was added to dissolve the antibiotic, followed by 200 μl of 0.01 M hydrochloric acid. The pH of the resulting solution was pH 8. An intravenous preparation of ciprofloxacin (Ciproxin 2 mg/ml) was purchased from Bayer (Basingstoke, United Kingdom). Dosing regimens were determined using pharmacokinetic data generated from previous studies (Barnes et al., 2017 (link); Hamblin et al., 2017 (link)).
For the in vivo studies, a 15 mg/ml solution of Finafloxacin was prepared by adding 2.1 ml of 0.01 M Tris buffer to 44 mg of Finafloxacin (containing 37.5 mg of active ingredient). 200 μl of 1 M sodium hydroxide was added to dissolve the antibiotic, followed by 200 μl of 0.01 M hydrochloric acid. The pH of the resulting solution was pH 8. An intravenous preparation of ciprofloxacin (Ciproxin 2 mg/ml) was purchased from Bayer (Basingstoke, United Kingdom). Dosing regimens were determined using pharmacokinetic data generated from previous studies (Barnes et al., 2017 (link); Hamblin et al., 2017 (link)).
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