Zolav^®: a new antibiotic for the treatment of acne

Figures & data

Table 1 MIC (µg/mL) of Zolav^® and clindamycin (control) against P. acnes

Test strain	Strain ID	Zolav^®	Clindamycin
P. acnes	ATCC 11827	2	0.063
P. acnes	ATCC 29399	2	0.063
P. acnes	ATCC 6919	2	0.063

Abbreviations: P. acnes, Propionibacterium acnes; MIC, minimum inhibitory concentration.

Table 2 Cytotoxicity of Zolav^® and staurosporin (control) in cell lines

	Cell line	Species	GI50	TGI	LC50
Zolav^®	HUVEC	Human	>30 µM	>30 µM	>30 µM
	Lung A549	Human	>30 µM	>30 µM	>30 µM
	Lung NCI-H460	Human	>30 µM	>30 µM	>30 µM
Staurosporin	HUVEC	Human	<1.0 nM	11.0 nM	0.023 µM
	Lung A549	Human	2.0 nM	0.052 µM	0.17 µM
	Lung NCI-H460	Human	8.0 nM	0.3 µM	0.61 µM

Abbreviations: GI₅₀, growth inhibition in 50% concentration; TGI, total growth inhibition; LC₅₀, lethal concentration of 50%; HUVEC, human umbilical vein endothelial cells.

Table 3 Activity of Zolav^® on revertant colonies of Escherichia coli in the presence of S9

Treatment	Concentration	S9	Revertant colonies/plate (n=3)
			Escherichia coli (ATCC 49979)
			Individual			Mean ± SD
Spontaneous revertant	–	(+)	33	24	36	31±6
DMSO (solvent)	100 µg/plate	(+)	34	33	32	33±1
2-Anthramine	30 µg/plate	(+)	752	751	758	754±4*
Zolav^®	1,500 µg/plate	(+)	27	35	23	28±6
	150 µg/plate	(+)	29	29	27	28±1
	15 µg/plate	(+)	32	30	33	32±1
	1.5 µg/plate	(+)	29	28	34	30±3

Notes: Significant mutagenic activity (≥3× of control) denoted as (*). Number of revertant colonies/plate is shown for each individual plate. S9 – homogenate from the rat liver containing the cytosol and microsomes. Mean ± SD value for triplicate plates of each treatment was calculated and shown after the individual plate counts.

Abbreviations: ATCC, American Type Culture Collection; DMSO, dimethyl sulfoxide; SD, standard deviation.

Figure 1 Propionibacterium acnes CFU per gram of skin tissue in mice.

Notes: Bars = mean + SEM; n=3 for “vehicle control 2 hours (baseline)” (dashed line) and n=5 for “vehicle control – 96 hours” and “Zolav^® – 96 hours 50 µg/mL (q8h)”.
Abbreviations: CFU, colony-forming unit; SEM, standard error of the mean; h, hours; q8h, every 8 hours.

Figure S1 Comparison of infection area between treated and untreated mice.

Notes: Pictures of mice from the untreated group (A and B) and Zolav^® at 50 µg/mL q8h (C and D) treated group at 26 hours (A and C) and at 50 hours (B and D) after challenge.

Table S1 Activity of Zolav^® on revertant colonies of Escherichia coli in the absence of S9

Treatment	Concentration	S9	Revertant colonies/plate (n=3)
			Escherichia coli (ATCC 49979)
			Individual			Mean ± SD
Spontaneous revertant	–	(−)	32	33	28	31±3
DMSO (solvent)	100 µg/plate	(−)	33	30	26	30±3
K2Cr2O7	30 µg/plate	(−)	146	147	142	145±3*
Zolav^®	1,500 µg/plate	(−)	25	29	29	28±3
	150 µg/plate	(−)	35	29	21	28±7
	15 µg/plate	(−)	23	32	30	28±4
	1.5 µg/plate	(−)	34	33	36	34±2

Notes: Significant mutagenic activity (≥3× of control) denoted as (*). Number of revertant colonies/plate is shown for each individual plate. S9 – homogenate from the rat liver containing the cytosol and microsomes. Mean ± SD value for triplicate plates of each treatment was calculated and shown after the individual plate counts.

Abbreviations: ATCC, American Type Culture Collection; DMSO, dimethyl sulfoxide; SD, standard deviation.

Table S2 Summary of Propionibacterium acnes intradermal infection model in mice in CFU/g (log10)

Mouse	Vehicle control – 2 hours (baseline)	Vehicle control – 96 hours	Zolav^® 50 μg/mL (q8h) – 96 hours
1	4.56	5.51	4.46
2	4.60	5.46	4.87
3	6.23	6.54	5.00
4		6.78	4.88
5		7.81	5.50
Mean	5.78	7.17	5.07
SD	0.956	0.979	0.374
SEM	0.552	0.438	0.167

Abbreviations: SEM, standard error of the mean; SD, standard deviation; q8h, every 8 hours.