Nitric Oxide Generation from Endogenous Substrates Using Metal-Organic Frameworks: Inclusion within Poly(vinyl alcohol) Membranes To Investigate Reactivity and Therapeutic Potential

摘要

Cu-BTTri (H(3)BTTri = 1,3,5-tris[1H-1,2,3-triazol-5-yl]benzene) is a water-stable, copper-based metal-organic framework (MOF) that exhibits the ability to generate therapeutic nitric oxide (NO) from S-nitrosothiols (RSNOs) available within the bloodstream. Immobilization of Cu-BTTri within a polymeric membrane may allow for localized NO generation at the blood-material interface. This work demonstrates that Cu-BTTri can be incorporated within hydrophilic membranes prepared from poly(vinyl alcohol) (PVA), a polymer that has been examined for numerous biomedical applications. Following immobilization, the ability of the MOF to produce NO from the endogenous RSNO S-nitrosoglutathione (GSNO) is not significantly inhibited. Poly(vinyl alcohol) membranes containing dispersions of Cu-BTTri were tested for their ability to promote NO release from a 10 mu M initial GSNO concentration at pH 7.4 and 37 degrees C, and NO production was observed at levels associated with antithrombotic therapeutic effects without significant copper leaching (<1%). Over 3.5 +/- 0.4 h, 10 wt % Cu-BTTri/PVA membranes converted 97 +/- 6% of GSNO into NO, with a maximum NO flux of 0.20 +/- 0.02 nmol.cm(-2).min(-1). Furthermore, it was observed for the first time that Cu-BTTri is capable of inducing NO production from GSNO under aerobic conditions. At pH 6.0, the NO-forming reaction of 10 wt % Cu-BTTri/PVA membrane was accelerated by 22%, while an opposite effect was observed in the case of aqueous copper(II) chloride. Reduced temperature (20 degrees C) and the presence of the thiol-blocking reagent N-ethylmaleimide (NEM) impair the NO-forming reaction of Cu-BTTri/PVA with GSNO, with both conditions resulting in a decreased NO yield of 16 +/- 1% over 3.5 h. Collectively, these findings suggest that Cu-BTTri/PVA membranes may have therapeutic utility through their ability to generate NO from endogenous substrates. Moreover, this work provides a more comprehensive analysis of the parameters that influence Cu-BTTri efficacy, permitting optimization for potential medical applications.