A convenient and rapid in vitro model to screen
steroid 5 alpha-reductase inhibitors, which are effective in the treatment of
benign prostatic hyperplasia (BPH), was developed. In the presence of
nicotinamide adenine dinucleotide phosphate (
NADPH),
steroid 5 alpha-reductase converts
testosterone to
dihydrotestosterone (DHT) which is a major etiologic factor of BPH.
NADPH has characteristic absorbance at 340 nm, and the absorbance spectrum may be used to identify
NADPH as a kind of the substrate in this enzymatic reaction. In this paper,
NADPH,
steroid 5 alpha-reductase, series concentration of
testosterone and
finasteride, and 4 ml Tris-HCl
buffer were continuously incubated together at 37 degrees C and the
NADPH OD values were continually measured. The descending rate of
NADPH OD340nm value by linear regression from the beginning to the 10th minute is close to the initial velocity of the enzymatic reaction. The precise activity of the
steroid 5 alpha-reductase was the slope after subtracting that of the blank control. The inhibition constant (Ki) of
steroid 5 alpha-reductase inhibitors could be calculated according to the Lineweaver-Burk plots. Two
drug screening models, the most common
isotope model and the novel model, were compared in this paper. The result showed that the latter one is more economical, quicker and more effective than the former one.