Evaluation of amino acid-mustard transport as L-type amino acid transporter 1 (LAT1)-mediated alkylating agents.

The L-type amino acid transporter 1 (LAT1, SLC7A5) is an Na+-independent neutral amino acid transporter the expression of which is located in retinal endothelial cells. Due to its broad substrate selectivity, LAT1 has been proposed to mediate the transport of amino acid-related drugs across the blood-tissue barriers. Here, we have investigated the transport screening of amino acid-mustards using a conditionally immortalized rat retinal capillary endothelial cell line (TR-iBRB2) which expresses LAT1. We synthesized 5 amino acid-mustards: tyrosine-mustard, phenylglycine-mustard, alanine-mustard, ornithine-mustard, and lysine-mustard. LAT1-mediated [3H]L-phenylalanine (Phe) uptake by TR-iBRB2 cells was inhibited in a competitive manner by tyrosine-mustard and phenylglycine-mustard as well as melphalan (phenylalanine-mustard). Phenylglycine-mustard was able to induce the efflux of [3H]Phe preloaded into the TR-iBRB2 cells expressing LAT1 through the obligatory exchange mechanism, although tyrosine-mustard, alanine-mustard, ornithine-mustard, lysine-mustard, and melphalan did not induce any significant efflux. These findings suggest that phenylglycine-mustard is a better substrate for LAT1 than melphalan and other amino acid-mustards.

Comparative study on the combined effects of bunazosin and nipradilol or timolol on intraocular pressure in normotensive rabbits.

PURPOSE: To compare the ocular hypotensive effect of nipradilol and timolol in combination with bunazosin in rabbits. METHODS: The intraocular pressure (IOP) in normal rabbits was measured using an applanation pneumatonograph. Nipradilol, timolol, and bunazosin were instilled, individually or in combination, into the inferior conjunctival sac. RESULTS: Nipradilol (0.25%), timolol (0.5%), and bunazosin (0.01%) individually lowered IOP. The IOP-lowering effects of both nipradilol and timolol were significantly enhanced by the combined application of bunazosin (0.01%). In the presence of 5% timolol or 0.1% bunazosin, IOP was further lowered by the addition of nipradilol. The IOP-lowering effect of nipradilol was partly inhibited by pretreatment with c-PTIO (10 mM), a nitric oxide (NO)-trapping agent. CONCLUSIONS: The present study demonstrated that the IOP-lowering effects of nipradilol are due to beta- and alpha1-blocking and NO-donating actions, and bunazosin has an additive effect on the IOP-lowering effect of nipradilol or timolol.

Potent reduction of intraocular pressure by nipradilol plus latanoprost in ocular hypertensive rabbits.

The present study was performed to evaluate the intraocular pressure (IOP)-lowering effect of nipradilol in combination with latanoprost on ocular normotensive and hypertensive rabbits. IOP was measured using an applanation pneumatonograph under topical application of 0.4% oxybuprocaine hydrochloride for corneal anesthesia. Ocular hypertension was induced by injection of 0.1 ml hypertonic saline (5% NaCl) into the vitreous body. Saline, nipradilol, latanoprost, sodium nitroprusside (SNP) or indomethacin was then instilled just after 5% NaCl injection. All drugs were instilled in the inferior conjunctival sac, using 50 microl drops. If more than two drugs were used, they were applied 5 min apart. Nipradilol lowered IOP in both ocular normotensive rabbits and ocular hypertensive rabbits, whereas latanoprost did not lower IOP in either. When nipradilol was applied in combination with latanoprost, the reduction in ocular hypertension was significantly enhanced, compared to the effect of nipradilol alone. A significantly potent reduction in ocular hypertension was also observed by the SNP-latanoprost combination. The IOP-lowering effects of SNP in combination with latanoprost were abolished by treatment with indomethacin. These results indicate that the IOP-lowering effect of latanoprost was enhanced when applied in combination with nipradilol or SNP, both of which have nitric oxide (NO)-donating actions. Since both combined effects were abolished by treatment with indomethacin, the mechanisms by which nipradilol combined with latanoprost lowered ocular hypertension may be related, at least in part, to the production of prostaglandins via the NO-donating action of nipradilol.