Binding of the beta-blockers timolol and H 216/44 to ocular melanin

The eyes from pigmented rabbits were instilled with the beta-adrenoceptor antagonists H 216/44 or timolol. After a single instillation (1.9 mumol), the iris and ciliary body contained H 216/44, which decreased with a half-life of approx. 43 days. Daily instillation caused a gradual increase in the content of H 216/44 and timolol in the iris and ciliary body, the steady-state concentration of timolol being 10 times higher than that of H 216/44. The concentrations of H 216/44 were seven times higher in the iris and ciliary body of pigmented rabbits than in albino animals. H 216/44 was reversibly bound to the melanosomes from the iris and ciliary body and not metabolized in this tissue. In vitro binding of timolol and H 216/44 to bovine melanosomes showed comparable multi-site binding curves. The binding of chlorpromazine was substantially higher. The beta-blockers could be more readily released from the melanosomes with aqueous solutions of salt and ethanol than with distilled water. It is concluded that both H 216/44 and timolol bind reversibly to ocular melanin. The differences in binding characteristics in vitro may only partly explain the differences in the in vivo binding to ocular melanin in the rabbit eye.