Discovery of GZ-793A, a novel VMAT2 inhibitor and potential pharmacotherapy for methamphetamine abuse.

摘要

Methamphetamine abuse is a serious public health concern affecting millions of people worldwide, and there are currently no viable pharmacotherapies to treat methamphetamine abuse. Methamphetamine increases extracellular dopamine (DA) concentrations through an interaction with the DA transporter (DAT) and the vesicular monoamine transporter-2 (VMAT2), leading to reward and abuse. While numerous studies have focused on DAT as a target for the discovery of pharmacotherapies to treat psychostimulant abuse, these efforts have been met with limited success. Taking into account the fact that methamphetamine interacts with VMAT2 to increase DA extracellular concentrations; the focus of the current work was to develop novel compounds that interact with VMAT2 to inhibit the effects of methamphetamine. Lobeline, the principal alkaloid found in Lobelia inflata, inhibits VMAT2 binding and function. Inhibition of VMAT2 was hypothesized to be responsible for the observed lobeline-induced inhibition of methamphetamine-evoked DA release in striatal slices and decrease in methamphetamine self-administration in rats. Lobeline has recently completed Phase Ib clinical trials demonstrating safety in methamphetamine abusers. Lobeline is also a potent inhibitor of nicotinic acetylcholine receptors (nAChRs), limiting selectivity for VMAT2. Chemical defunctionalization of the lobeline molecule afforded analogs, meso-transdiene (MTD) and lobelane, which exhibited decreased affinity for nAChRs. MTD, an unsaturated analog of lobeline, exhibited similar affinity for VMAT2 and increased affinity for DAT compared to lobeline. Conformationally-restricted MTD analogs exhibited decreased affinity for DAT compared to MTD, while retaining affinity at VMAT2. One analog, UKMH-106 exhibited high affinity and selectivity for VMAT2 and inhibited METH-evoked DA release from striatal slices. Unfortunately, the MTD analogs exhibited poor water solubility which limited further investigation of these promising analogs. Importantly lobelane, a saturated analog of lobeline, exhibited increased affinity and selectivity for VMAT2 compared to lobeline. To improve water solubility, a N-1,2-dihydroxypropyl (diol) moiety was incorporated into the lobelane molecule. GZ-793A, an N-1,2-diol analog, potently and competitively inhibited VMAT2 function, exhibiting over 50-fold selectivity for VMAT2 over DAT, serotonin transporters and nAChRs. GZ-793A released DA from preloaded synaptic vesicles, fitting a two-site model with the high-affinity site inhibited by tetrabenazine and reserpine (classical VMAT2 inhibitors), suggesting a VMAT2-mediated mechanism of release. Further, low concentrations of GZ-793A that selectively interact with high-affinity sites on VMAT2 to evoke DA release, inhibit methamphetamine-evoked DA release from synaptic vesicles. Results showed that increasing concentrations of GZ-793A produced a rightward shift in the METH concentration response; however, the Schild regression revealed a slope different from unity, consistent with surmountable allosteric inhibition. In addition, GZ-793A specifically inhibited methamphetamine-evoked DA release in striatal slices and methamphetamine self-administration in rats. To examine the possibility that GZ-793A produced DA depletion, the effect of a behaviorally active dose of GZ-793A on DA content in striatal tissue and striatal vesicles was determined. GZ-793A administration did not alter DA content in striatal tissue or vesicles and pretreatment with GZ-793A prior to methamphetamine administration did not exacerbate the DA depleting effects of methamphetamine. Importantly, GZ-793A was shown to protect against methamphetamine-induced striatal DA depletions. Thus, GZ-793A represents an exciting new lead in the development of pharmacotherapies to treat methamphetamine abuse.;KEYWORDS: Lobeline, Methamphetamine, Vesicular Monoamine Tranporter-2, Drug Discovery, GZ-793A.