Identification of human dopamine D1-like receptor agonist using a cell-based functional assay1

Jiang, Nan; Ouyang, Keqing; Cai, Shaoxi; Hu, Yinghe; Zhou, Xu

doi:10.1111/j.1745-7254.2005.00199.x

Cited by 6 publications

(5 citation statements)

References 27 publications

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“…The number of hits as a percentage of the library (1.8%) was comparable to other published screens [36], [37], [38], suggesting that the assay was specific with a low number of false positives. Nevertheless, we had set the initial screen threshold of 20% upregulation low enough to avoid false negatives, in the expectation that some of the initial hits would be false positives due to ‘statistical noise’.…”

Section: Discussionsupporting

confidence: 78%

Drug Discovery for Duchenne Muscular Dystrophy via Utrophin Promoter Activation Screening

et al. 2011

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BackgroundDuchenne muscular dystrophy (DMD) is a devastating muscle wasting disease caused by mutations in dystrophin, a muscle cytoskeletal protein. Utrophin is a homologue of dystrophin that can functionally compensate for its absence when expressed at increased levels in the myofibre, as shown by studies in dystrophin-deficient mice. Utrophin upregulation is therefore a promising therapeutic approach for DMD. The use of a small, drug-like molecule to achieve utrophin upregulation offers obvious advantages in terms of delivery and bioavailability. Furthermore, much of the time and expense involved in the development of a new drug can be eliminated by screening molecules that are already approved for clinical use.Methodology/Principal FindingsWe developed and validated a cell-based, high-throughput screening assay for utrophin promoter activation, and used it to screen the Prestwick Chemical Library of marketed drugs and natural compounds. Initial screening produced 20 hit molecules, 14 of which exhibited dose-dependent activation of the utrophin promoter and were confirmed as hits. Independent validation demonstrated that one of these compounds, nabumetone, is able to upregulate endogenous utrophin mRNA and protein, in C2C12 muscle cells.Conclusions/SignificanceWe have developed a cell-based, high-throughput screening utrophin promoter assay. Using this assay, we identified and validated a utrophin promoter-activating drug, nabumetone, for which pharmacokinetics and safety in humans are already well described, and which represents a lead compound for utrophin upregulation as a therapy for DMD.

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Section: Discussionsupporting

confidence: 78%

Drug Discovery for Duchenne Muscular Dystrophy via Utrophin Promoter Activation Screening

et al. 2011

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“…The D 5 receptor appears to exhibit higher affinity for dopamine than the D 1 receptor when transiently or stably expressed in COS-7 or Ltk- cells (Jarvie et al, 1993). More recently, Jiang and colleagues (Jiang et al, 2005) reported on a number of compounds isolated from nature that exhibit different affinities for the D 1 and D 5 dopamine receptors. For instance, SBG492, one of the phytochemicals screened, exhibited an EC50 of 343 μg/ml at the D 1 receptor while the value for the D 5 receptor was 32 μg/ml.…”

Section: Introductionmentioning

confidence: 99%

Pharmacology of signaling induced by dopamine D1-like receptor activation

Undieh

2010

Pharmacology & Therapeutics

107

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Dopamine D1-like receptors consisting of D1 and D5 subtypes are intimately implicated in dopaminergic regulation of fundamental neurophysiologic processes such as mood, motivation, cognitive function, and motor activity. Upon stimulation, D1-like receptors initiate signal transduction cascades that are mediated through adenylyl cyclase or phosphoinositide metabolism, with subsequent enhancement of multiple downstream kinase cascades. The latter actions propagate and further amplify the receptor signals, thus predisposing D1-like receptors to multifaceted interactions with various other mediators and receptor systems. The adenylyl cyclase response to dopamine or selective D1-like receptor agonists is reliably associated with the D1 subtype, while emerging evidence indicates that the phosphoinositide responses in native brain tissues may be preferentially mediated through stimulation of the D5 receptor. Besides classic coupling of each receptor subtype to specific G proteins, additional biophysical models are advanced in attempts to account for differential subcellular distribution, heteromolecular oligomerization, and activity-dependent selectivity of the receptors. It is expected that significant advances in understanding of dopamine neurobiology will emerge from current and anticipated studies directed at uncovering the molecular mechanisms of D5 coupling to phosphoinositide signaling, the structural features that might enhance pharmacological selectivity for D5 versus D1 subtypes, the mechanism by which dopamine may modulate phosphoinositide synthesis, the contributions of the various responsive signal mediators to D1 or D5 interactions with D2-like receptors, and the spectrum of dopaminergic functions that may be attributed to each receptor subtype and signaling pathway.

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“…10−12 Dysregulation of the catecholamine dopamine plays diverse roles in pathogenesis and therapy for different diseases such as schizophrenia, Parkinson's disease (PD), drug addiction, and bipolar disorders. 13−15 In the preceding few decades, several test systems have been developed to characterize binding properties or functionality of ligands concerning either the D 1 R or D 2 R. 7,16,17 Unfortunately, the other subtypes have not been as much of a focus.…”

mentioning

confidence: 99%

“…G protein-coupled receptors (GPCRs) are the most studied targets in drug discovery and are the most common transmembrane receptors in the mammalian genome. − GPCRs are connected to heterotrimeric proteins containing α, β, and γ subunits and can be classified according to their primary coupling behavior to the Gα subunit, which is divided into α s -, α i/o -, α q/11 -, and α 12/13 families. , The activation of a GPCR subsequently regulates intracellular signaling by engaging heterotrimeric G proteins or β-arrestins, which transfer the signal to further downstream effector proteins or lead to internalization and desensitization of the receptor. − …”

mentioning

confidence: 99%

“…In the preceding few decades, several test systems have been developed to characterize binding properties or functionality of ligands concerning either the D 1 R or D 2 R. ,, Unfortunately, the other subtypes have not been as much of a focus. Selectivity studies between all five dopamine receptors, coupling studies contrary to primary coupling of dopamine receptors, and the monitoring of their functional response all remain under-researched .…”

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confidence: 99%

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Activation of Multiple G Protein Pathways to Characterize the Five Dopamine Receptor Subtypes Using Bioluminescence Technology

Mönnich,

Humphrys,

Höring

et al. 2024

ACS Pharmacol. Transl. Sci.

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G protein-coupled receptors show preference for G protein subtypes but can recruit multiple G proteins with various downstream signaling cascades. This functional selection can guide drug design. Dopamine receptors are both stimulatory (D 1 -like) and inhibitory (D 2 -like) with diffuse expression across the central nervous system. Functional selectivity of G protein subunits may help with dopamine receptor targeting and their downstream effects. Three bioluminescence-based assays were used to characterize G protein coupling and function with the five dopamine receptors. Most proximal to ligand binding was the miniG protein assay with split luciferase technology used to measure recruitment. For endogenous and selective ligands, the G-CASE bioluminescence resonance energy transfer (BRET) assay measured G protein activation and receptor selectivity. Downstream, the BRET-based CAMYEN assay quantified cyclic adenosine monophosphate (cAMP) changes. Several dopamine receptor agonists and antagonists were characterized for their G protein recruitment and cAMP effects. G protein selectivity with dopamine revealed potential G q coupling at all five receptors, as well as the ability to activate subtypes with the "opposite" effects to canonical signaling. D 1 -like receptor agonist (+)-SKF-81297 and D 2 -like receptor agonist pramipexole showed selectivity at all receptors toward G s or G i/o/z activation, respectively. The five dopamine receptors show a wide range of potentials for G protein coupling and activation, reflected in their downstream cAMP signaling. Targeting these interactions can be achieved through drug design. This opens the door to pharmacological treatment with more selectivity options for inducing the correct physiological events.

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Identification of human dopamine D1-like receptor agonist using a cell-based functional assay1

Cited by 6 publications

References 27 publications

Drug Discovery for Duchenne Muscular Dystrophy via Utrophin Promoter Activation Screening

Drug Discovery for Duchenne Muscular Dystrophy via Utrophin Promoter Activation Screening

Pharmacology of signaling induced by dopamine D1-like receptor activation

Activation of Multiple G Protein Pathways to Characterize the Five Dopamine Receptor Subtypes Using Bioluminescence Technology

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