Aryl Phosphate Derivatives of Bromo-Methoxy-Azidothymidine: MATERIALS AND METHODS(3)
Sperm Immobilization Assay (SIA)
To evaluate the spermicidal effects of AZT and WHI compounds 01, 02, 03, 04, 05, 06, 07, 08, 09, 10, 11, and 12 in comparison with the detergent spermicide N-9 (IG-EPAL CO-630; Rhone Poulenc, Cranbury, NJ), a highly motile fraction of pooled donor sperm (n = 10) was prepared by discontinuous (90-45%) Percoll gradient (Conception Technologies, San Diego, CA) centrifugation and the ‘‘swim-up’’ method as previously described. All donor specimens were obtained after informed consent and in compliance with the guidelines of the Wayne Hughes Institute Institutional Review Board. Motile sperm (> 10 X 106/ml) were suspended in 0.5 ml of Biggers, Whitten, and Whittingam’s medium (BWW) containing 25 mM He-pes (Irvine Scientific, Santa Ana, CA) and 0.3% BSA (fraction V; Sigma Chemical Co., St. Louis, MO) in the presence and absence of serial 2-fold dilutions of test substance (300-4.6 ^M) in 1% dimethyl sulfoxide (DMSO). buy levaquin online
The stock solutions of synthetic WHI compounds were prepared in DMSO (10 mg/ml) and diluted in DMSO to yield the desired concentrations. A corresponding volume of DMSO (1%) was added to control tubes. N-9 was diluted in BWW-0.3% BSA (pH 7.4) to yield the desired concentrations (300-4.6 ^M). After 3 h of incubation at 37°C, the percentage of motile sperm was evaluated by computer-assisted sperm motion analysis (CASA) as described previously. The percentage motilities were compared with those of sham-treated control suspensions of motile sperm. The spermicidal activity of test compounds was expressed as the EC50 values (the final concentration of the compound in medium that decreases the proportion of motile sperm by 50%).