Comparison of cell-permeable calpain inhibitors and E64 in reduction of cataract in cultured rat lenses.
E64, an inhibitor of calpain (EC 3.4.22.17) and other cysteine proteases, slows the rate of formation of cataract in cultured rat lenses. The purpose of this study was to determine (1) why E64, a charged compound with little cell permeability, was effective in reducing cataract in cultured lens and (2) whether uncharged more permeable protease inhibitors are more effective than E64 in preventing cataract. Results showed that E64 entered the lens, but only after the lens was treated with the calcium ionophore, A23187, or sodium selenite, both of which cause cataracts. Therefore, the uptake and subsequent effectiveness of E64 may be related to a generalized increase in membrane permeability during induction of cataract in culture. Three protease inhibitors, reported to have improved cell permeability, were compared with E64 for their ability to prevent cataracts in cultured lenses. cBz-ValPheH, calpain inhibitors I and II, are uncharged-aldehyde inhibitors of calpain. Calpain inhibitors I and II even at high concentrations were not effective at reducing lens opacity caused by calcium ionophore and were toxic to the lens. cBz-ValPheH, which is slightly toxic to the lens, was able to significantly reduce lens opacity induced by calcium ionophore. The presented data suggest that while E64 decreases cataract formation in cultured lens, the more cell permeable inhibitor, cBz-ValPheH, may have greater efficacy as an anticataract drug in vivo.
Lampi
K J
KJ
Department of Biochemistry, Oregon Health Sciences University, Portland 97201.
Kadoya
K
K
Azuma
M
M
David
L L
LL
Shearer
T R
TR
D000595:Amino Acid Sequence; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002154:Calpain / Q000037:antagonists & inhibitors*; D002386:Cataract / Q000139:chemically induced / Q000201:enzymology / Q000517:prevention & control*; D002463:Cell Membrane Permeability / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D046508:Culture Techniques; D015853:Cysteine Proteinase Inhibitors / Q000493:pharmacokinetics / Q000627:therapeutic use* / Q000633:toxicity; D004151:Dipeptides / Q000493:pharmacokinetics / Q000627:therapeutic use / Q000633:toxicity; D004347:Drug Interactions; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000493:pharmacokinetics / Q000627:therapeutic use* / Q000633:toxicity; D007908:Lens, Crystalline / Q000187:drug effects / Q000378:metabolism; D007930:Leucine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000627:therapeutic use / Q000633:toxicity; D007976:Leupeptins / Q000493:pharmacokinetics / Q000627:therapeutic use / Q000633:toxicity; D008969:Molecular Sequence Data; D051381:Rats; D017207:Rats, Sprague-Dawley
D015853:Cysteine Proteinase Inhibitors; D004151:Dipeptides; D006023:Glycoproteins; D007976:Leupeptins; C039304:calpain inhibitors; D000001:Calcimycin; D002154:Calpain; D007930:Leucine; C032854:leupeptin; C024974:E 64; C058076:calpain inhibitor III