Implications for the therapeutic treatment of impulsivity and executive dysfunction with dopaminergics - Stereoselective D1 and D2 DA.

R ligands
Federico dOleire
1, 2 Uquillas 1. Helen Wills Neuroscience Institute, 2. Molecular and Cell Biology Department - University of California, Berkeley
Review of the D1 DAR
The D1 receptor is an abundant metabotropic G-protein-coupled receptor that activates excitatory processes in the intracellular space of the postsynaptic neuron. It stimulates adenylyl cyclase, producing cAMP and protein kinases in the process, both of which are involved in gene transcription and modulation of the D2 receptor complex (1). Can be found in the hypothalamus, cerebral cortex, hippocampus, striatum, olfactory bulb, retina, the entire eye, trachea, and white adipose tissue (2). D1 receptors play a role in working memory and prefrontal cortex (PFC) impairment (3), and there have been approximately 300 binding assays conducted on the D1 receptor within the past four years, with more than 2,000 ligand assays with associated bioactivity (4). This new wave of research has revealed lots of potential for D1 ligands.

Impulsivity & Decision Making

Impulsivity, a feature of human behavior that can both be helpful and problematic, has been correlated with several psychopathologies characterized by poor inhibitory control, such as substance use disorders and anxiety disorders (12).
HUMAN D1 RECEPTOR The Human Protein Atlas

Review of the D2 DAR

A G-protein coupled receptor that inhibits activity via the Gi cascade (19). Found in the cerebellum, brainstem, hypothalamus, cerebral cortex, hippocampus, striatum, olfactory bulb and epithelium, retina, the entire eye, the vena cava, heart atrium and ventricle, testes, and in robust densities in the pituitary gland (20). Its localization in the striatum is denser than in the cortex as well, and it has been particularly implicated in the indirect striato-pallido-cortical circuitry thought to mediate inhibitory control and appropriate decision-making (21). D2 receptor abnormalities in the dorsal striatum, mPFC, and orbitofrontal cortex (OFC) also correlate with increased risk-taking behavior (22). HUMAN D2 RECEPTOR The Human Protein Atlas

Custom tailoring for the D1 DAR

D1 receptor density in the caudate nucleus, is positively correlated with dorsololateral PFC (DL-PFC) connectivity to the right parietal cortex, a network implicated in maintenance of information in working memory processes (5). On the other hand, an abundance of D1 mRNA expression in the nucleus accumbens is correlated with high impulsive action in animal models, and D1 expression in the medial PFC (mPFC) has been correlated with decision-making where too much or too little D1 expression can cause dysfunction in the system (6,7). This over- or under-stimulation of the PFC can cause impairment in an inverted u-shape curve fashion (8). D1 receptor antagonists like SCH-23390 seem to increase impulsive decision-making in rats during delayed-reward tasks (9). A D1 agonist like SKF 38393 on the other hand has showed improved accuracy and decreased impulsivity in rats when engaged in a five-choice serial reaction time task (5CSRT) (10). Findings like these also corroborate an upside down u-shape curve that governs DAs effects on working memory, where high doses of D1 DAR agonists induce impulsive decision-making behavior.

With more and more evidence that working memory performance is correlated with the ability to make thoughtful and deliberate decisions, pharmacological interventions that could enhance decision-making and working memory performance would greatly benefit many. In particular, those suffering from addictions (13,14).

Fig.1 - Adapted by Winstanley, from Ainslie, 1975. Used with permission of the author.

Custom tailoring for the D2 DAR

In monkeys, D2 receptor expression in the brain rapidly decreases by about 15-20% after cocaine selfadministration, and remains underexpressed by about 20% even after one year following exposure (23). This overall decrease in D2 activity seen in monkeys has also been demonstrated in the striatum [Fig.2] and PFC of human cocaine abusers recently by Goldstein, et al. Hypometabolism in areas known to be involved with inhibitory control might contribute to poor decision-making, and could more easily lead to addicted states (24). Stereoselectivity for the D2 receptor in particular, was first examined in the 1990s: Studies on the enantiomers of 11-hydroxy-N-allyl and 10, 11-dihydroxy-N-allyl revealed enantiomeric affinity for the D2 receptor in the striatum of rat brain tissue (25). By discovering an effective base for the development of radio-labeled ligands, this study spurred much further research into DAR subtype stereoselective agonists and antagonists.

For example, a recent study on impulsivity showed that the emotionality and immediacy components of a reward influence decision-making more so in adolescents who binge drink during adolescence (15). The group of adolescents showed heightened BOLD responses in the amygdala and insula as well, and had a worse score on the Iowa Gambling Task than control groups. Strengthening functionality in regions like these could translate into positiive behavioral outcomes
Identifying brain regions associated with impulsivity and cognitive control may allows us to fine-tune our research for experimental therapeutics, but it is only the first step in understanding the pharmacokinetic profile of drugs seen to enhance executive function.

Pfeiffers Rule - Eutomers & Distomers

Enantiomers of doxanthrine (DOX), a potent and selective D1 agonist synthesized by David Nichols at Purdue University and used as a therapeutic agent for treating Parkinsons, were examined for pharmacological potency (16). They found that the eutomer - the more potent stereoisomer - of the racemic DOX, (+)-DOX, had not only greater intrinsic potency for the D1 receptor than its distomer, (-)-DOX, but (-)-DOX actually decreased the efficacy of both dopamine and (+)-DOX. In cloned human D1 receptors, (+)-DOX has an EC50 (effective concentration for 50% of the sample to experience effects) for enhancing cAMP accumulation of 29nM whereas (-)-DOX has a much larger EC50 (1,000nM) and is found to not even be an exclusive agonist at D1 receptors (17). In fact, (-)-DOX had comparable potencies as Clonidine at 2c-adrenergic receptors. Some in vivo findings have found however that while intraperitoneal administration of (+)-DOX has longer lasting effects than (-)-DOX, the latter has greater activity after oral administration in rats (18). In comparison, these favor (-)-DOX as a candidate for orally administered D1 ligand research. Lastly, a known obstacle with most D1 DAR therapeutics is that they appear to not be as effective with oral routes of administration in humans, so this in vivo finding must be further explored.

Insight into improving ligand binding

The David Nichols laboratory recently discovered that there are a number of transmembrane receptor domains on the D1 receptor, which engage its catechol agonists via polar interactions (11). The bioactive transmembrane domains on the D1 receptor could very well be an effective new target for D1 ligands if we can better understand more about their implications for ligand binding. These may be further studied using already characterized ligands, and might prove helpful for the development of better pharmacological treatments for the D1 DAR system.
DA pharmacotherapy improves with stereo-chemical insight
The discovery of dopamine stabilizers was one of the many innovations that came from the development of tritated dopamine bases for binding and displacement assays. (4-[3-(methylsulfonyl) phenyl]-1-propylpiperidine, a member of the pridopidine family and a D2 antagonist, was found to have a relatively high Ki (inhibition constant) when displacing characterized DA ligands (30). The low affinity profile of 4-[3-(methylsulfonyl) phenyl]-1-propylpiperidine allows it to bind to the D2 DAR only when in its open conformation, allowing the receptor to quickly regain responsiveness. This feature may promote its dopaminergic stabilizing and agonist-like qualities.
Acknowledgements
The authors would like to acknowledge the Molecular and Cell Biology Depatment at the University of California, Berkeley, and the support of Dr. David Presti for his continued support with this work. The content presented here is solely the responsibility of the authors and does not necessarily represent the official views of the University of California.

D2 antagonists like raclopride have been shown to increase reaction times and decrease overall cognitive function in rat models, an effect reversed by the indirect DA agonist, d-amphetamine (26). Raclopride may also increase feeding bouts in rats upon its administration to the nucleus accumbens.

[Fig.2] Lower Striatal Dopamine D2 Receptor Binding in Drug Users During Withdrawal From Cocaine, Meth-amphetamine, and Alcohol Than in Normal Comparison Subjects (Goldstein, 2002). Used with permission from the author.

A recent comparative study between D1 and D2 modulation showed that activation of the D2 DAR complex may attenuate risk-taking in rats, in comparison with activation of the D1 DAR (27; 28). The battery of studies conducted on the D2 DAR complex and particularly the striatum show great potential for the development of effective therapeutics geared towards improving poor decision-making (29).

References
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Take Home Message The future for addiction treatment and cognitive enhancers will not only be driven by the continued development of stereo-selective ligands tailored for desired efficacies for different receptor subtypes and localities implicated in poor inhibitory control, but also by the elucidation of the mechanisms in which these receptors modulate and interact with each other. Some obstacles facing effective enantiomeric affinity include the interactions between DAR subtype densities in different localities in the cortex, along with single nucleotide polymorphisms to receptor structures, known to influence the efficacy of dopaminergic drugs. In addition, the effect of dopaminergic ligands may be governed by an inverted u-shape curve, where DA baseline levels predict working memory performance upon administration of a dopaminergic. Lastly, pharmacokinetic profiles of these and other therapeutics may be explored by examining their Ki (inhibition constant) values to reveal affinity profiles for its own, and other receptors, reducing side effects, and improving desired outcomes.