Fig 1.
EP012 and EP055 interaction with Eppin.
(A) AlphaScreen assay of a-EAb binding to EPPIN, closed symbols are AlphaScreen assay, open symbols are for non-specific interactions of compound with assay components. EP012 = box/blue line, EP055 = circle/red line. (B) CASA Assay showing the relative motility index (RMI = straight-line velocity (VSL) x % normal motility normalized to DMSO vehicle control). EP012 = box/blue line, EP055 = circle/red line. Curves represent the mean ± SEM of 4 human sperm donors.
Fig 2.
EP055 time course and tissue levels in Macaca fascicularis after a single i.v. injection.
(A) Plasma half-life determination after infusion of a single dose (n = 3 males; 1mg/kg). t1/2 = 10.6 ± 2 minutes. (B) Testis tissue levels (n = 2 males per time point; 63.25 mg/kg) at 2 hr are statistically different from 2 hr epididymis levels (p < 0.05) by One-way ANOVA. The differences at 6 hr were not significant. There was no difference in the ratio of E/T at either time point. (C) Total μg of compound per tissue shows no significant difference between 2 hr levels and 6 hr levels (p < 0.05) by One-way ANOVA.
Fig 3.
Levels of EP055 in individual Macaca mulatta males after a single i.v. dose.
Each animal was given a low dose of EP055 (75–80 mg/kg). After sperm motility returned to normal, the same animal was given the high dose (125–130 mg/kg). Blue is male 28187, Orange is male 28043, Green is male 25854, Purple is male 28106. (A) Plasma and (C) Semen levels after the low dose of EP055. (B) Plasma and (D) Semen levels after the high dose of EP055.
Table 1.
EP055 levels in rhesus macaque (Macaca mulatta) plasma and semen after infusion.
Fig 4.
Effect of EP055 on rhesus macaque (Macaca mulatta) sperm parameters.
Closed circles represent percent normal motility and bars represent semen levels of EP055 in A and B. (A) Correlation between EP055 semen levels and normal sperm motility after a single low dose (75–80 mg/kg) # show the significant increase in EP055 semen levels compared to untreated (Kruskal-Wallis analysis, p < 0.05). (B) Correlation between EP055 semen levels and normal sperm motility after a single high dose (125–130 mg/kg) # show the significant increase in EP055 semen levels and * show the significant decreases in motility by Kruskal-Wallis analysis (p< 0.05, compared to untreated). (C) Photograph of sperm from male 28043 at 28–30 hrs after the high dose showing clumping. See S1 Movie. (D) Photograph of sperm from male 28043 at high dose recovery (18 days post-infusion) showing normal spacing at the same sperm density. See S2 Movie. Results shown are for the mean ± SEM of n = 4 animals. Fertile and sub-fertile indices of motility are based on human clinical parameters [20].