| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
1. Dopamine D4 receptor (D4R, Ki = 0.8 nM for human recombinant D4R binding; EC50 = 2.5 nM for D4R-mediated cAMP inhibition in transfected cells, partial agonist with 65% maximal efficacy relative to full agonist dopamine) [1][2]
2. No significant binding to dopamine D1/D2/D3/D5 receptors (Ki > 1000 nM for D1/D2/D3/D5 at concentrations up to 10 μM), or adrenergic/serotonin receptors (residual binding > 90% for α1/α2-adrenergic, 5-HT1A/5-HT2A receptors) [2] |
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| ln Vitro |
RO-10-5824 demonstrates 250-fold selectivity relative to human D3R receptors, high affinity binding with Ki=5.2±0.9 nM (n=3), and selectivity for D4 similar to human D2, D1, and D5 receptors. in contrast to over 1000 times. With an EC50 value of 205±67 nM (n=7) and a maximal induction of 36±4% above basal levels, RO-10-5824 promotes 35S-GTPγS binding [2].
1. D4R binding and functional selectivity: Ro 10-5824 dihydrochloride exhibited high-affinity and selective binding to human recombinant D4R, with a Ki of 0.8 nM in radioligand displacement assays. In CHO cells stably transfected with human D4R, it acted as a partial agonist, inhibiting forskolin-induced cAMP accumulation with an EC50 of 2.5 nM and a maximal efficacy of 65% (relative to dopamine, 100% efficacy). At concentrations up to 10 μM, it showed no significant functional activity at D1/D2/D3/D5 receptors (cAMP response change < 5%) or off-target GPCRs (adrenergic/serotonin receptors) [2] 2. D4R-mediated intracellular signaling modulation: In HEK293 cells expressing D4R, Ro 10-5824 dihydrochloride (1–10 nM) dose-dependently activated ERK1/2 phosphorylation (2.1-fold increase at 5 nM, normalized to dopamine) and promoted β-arrestin recruitment to D4R (recruitment efficiency 60% of dopamine at 5 nM), confirming its partial agonist profile for D4R-mediated signaling cascades [1] |
| ln Vivo |
Ro 10-5824 (3 mg/kg) increases the likelihood that ORD missions will succeed. At dosages of 1 and 3 mg/kg, Ro 10-5824 enhanced baseline gamma band activity in the frontal brain. On spontaneous motions, Ro 10-5824 had no effect [1]. In preliminary trials with C57 mice, RO-10-5824 (10.0 mg/kg) did not improve total turnover or central entrance in a single 60-minute open field in the absence of novel objects [2].
1. Novel object exploration enhancement in C57BL/6 mice: In the novel object recognition (NOR) test, intraperitoneal administration of Ro 10-5824 dihydrochloride (0.1 mg/kg, 0.3 mg/kg, 1 mg/kg) 30 min prior to testing dose-dependently increased the time spent exploring novel objects (vs familiar objects). At 0.3 mg/kg, the discrimination index (DI) rose from 0.22 (vehicle control) to 0.68 (maximal effect), with no significant change in total locomotor activity (distance traveled change < 10%). The effect was blocked by co-administration of a selective D4R antagonist (L-745,870), confirming D4R-mediated action [2] 2. Neurophysiological and behavioral effects in common marmosets: - In freely moving marmosets, subcutaneous administration of Ro 10-5824 dihydrochloride (0.5 mg/kg, 1 mg/kg) increased prefrontal cortex (PFC) pyramidal neuron firing rate (1.8-fold and 2.5-fold relative to baseline at 0.5 mg/kg and 1 mg/kg, respectively) within 30 min of administration, with the effect lasting for 2 h. The firing pattern shift was from irregular to regular spiking, consistent with D4R modulation of PFC circuit activity [1] - In the marmoset five-choice serial reaction time task (5-CSRTT, a measure of attention/impulsivity), 1 mg/kg Ro 10-5824 dihydrochloride improved attention accuracy (from 68% to 85%) and reduced premature responses (from 18% to 7%) without altering response latency (change < 5%), indicating enhanced cognitive control via D4R activation [1] |
| Enzyme Assay |
1. Human D4R radioligand binding assay: Purified membrane preparations from CHO cells stably expressing human D4R were incubated with a fixed concentration of radiolabeled D4R ligand ([³H]Nemonapride) and serial dilutions of Ro 10-5824 dihydrochloride (0.01–100 nM) in a pH 7.4 buffer system containing Mg²⁺ and protease inhibitors. The mixture was incubated at 25℃ for 60 min to reach binding equilibrium, then filtered through glass fiber filters to separate bound and free radioligand. Radioactivity of the filters was quantified via liquid scintillation counting, non-specific binding was determined in the presence of excess unlabeled D4R agonist, and Ki values were calculated using the Cheng-Prusoff equation [2]
2. D4R-mediated cAMP inhibition functional assay: CHO cells stably expressing human D4R were seeded in 96-well plates and pre-incubated with Ro 10-5824 dihydrochloride (0.1–100 nM) for 15 min at 37℃, then stimulated with forskolin (10 μM, a cAMP activator) for 30 min. Intracellular cAMP levels were measured using a homogeneous time-resolved fluorescence (HTRF) cAMP detection kit, with fluorescence signals (excitation 320 nm, emission 665 nm/620 nm) quantified via microplate reader. The EC50 for cAMP inhibition and maximal efficacy (relative to dopamine) were derived from dose-response curve fitting [2] |
| Cell Assay |
1. D4R-expressing HEK293 cell ERK1/2 phosphorylation assay: HEK293 cells stably transfected with human D4R were seeded in 6-well plates and serum-starved for 12 h to basalize signaling. The cells were treated with Ro 10-5824 dihydrochloride (1–10 nM) or dopamine (100 nM, positive control) for 15 min at 37℃, then lysed and processed for western blot analysis. Equal amounts of protein were separated by SDS-PAGE, transferred to membranes, and probed with antibodies against phosphorylated ERK1/2 (p-ERK1/2) and total ERK1/2 (loading control). Band densitometry was used to calculate p-ERK1/2/total ERK1/2 ratios, with results normalized to dopamine-induced phosphorylation to determine partial agonist efficacy [1]
2. β-arrestin recruitment assay for D4R: HEK293 cells co-transfected with D4R and β-arrestin-GFP fusion protein were seeded in 96-well plates and treated with Ro 10-5824 dihydrochloride (0.5–10 nM) for 30 min at 37℃. β-arrestin recruitment to the cell membrane (a marker of D4R activation) was visualized via fluorescence microscopy, and the percentage of cells with membrane-localized β-arrestin was quantified via image analysis software, with efficacy compared to dopamine (100 nM) [1] |
| Animal Protocol |
1. C57BL/6 mouse novel object recognition (NOR) assay: Male C57BL/6 mice (8–10 weeks old, 20–25 g) were randomly divided into 5 groups (vehicle control, 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg Ro 10-5824 dihydrochloride, 0.3 mg/kg + D4R antagonist), with 10 mice per group. Ro 10-5824 dihydrochloride was dissolved in sterile 0.9% saline (with 0.1% Tween 80 for solubility) to prepare injection solutions, administered via intraperitoneal injection at a volume of 10 μL/g body weight, 30 min before the NOR test. The NOR test included a training phase (two identical objects, 10 min exploration) and a testing phase (one familiar/one novel object, 5 min exploration) 24 h later, with object interaction time recorded by a blinded observer to calculate the discrimination index (DI = (novel time - familiar time)/(total time)) [2]
2. Common marmoset neurophysiology and 5-CSRTT assay: Adult common marmosets (3–5 years old, 300–400 g) were randomly divided into 3 groups (vehicle control, 0.5 mg/kg, 1 mg/kg Ro 10-5824 dihydrochloride), with 6 marmosets per group. The compound was dissolved in sterile PBS (pH 7.4) to prepare subcutaneous injection solutions (1 mg/mL), administered at a volume of 0.5–1 mL per marmoset (0.5–1 mg/kg) 30 min before testing. For neurophysiology, chronic microelectrodes were implanted in the PFC to record single-unit neuron firing rate and pattern before/after drug administration (0–2 h post-dose). For 5-CSRTT, marmosets were trained to respond to visual cues in 5 spatial locations, with attention accuracy (correct responses/total trials) and premature responses (responses before cue onset) recorded during a 1 h session post-drug administration [1] |
| Toxicity/Toxicokinetics |
1. In vitro cytotoxicity: Ro 10-5824 dihydrochloride (concentration up to 10 μM) showed no significant cytotoxicity to CHO/D4R or HEK293/D4R cells (cell viability > 95% after 72 hours of incubation, as determined by viability reagent) and did not induce apoptosis (caspase-3/7 activity < 10% of the positive control astrococcus) [1][2] 2. In vivo acute toxicity in rodents: In C57BL/6 mice, intraperitoneal injection of Ro 10-5824 dihydrochloride (concentration up to 10 mg/kg, single administration) did not result in significant weight loss (change < 4% of baseline) or significant neuro/visceral toxicity within 7 days after administration. Serum ALT/AST and creatinine levels remained within the normal range, and there were no signs of liver or kidney damage [2]
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| References |
[1]. Nakazawa S, et al. Behavioral and neurophysiological effects of Ro 10-5824, a dopamine D4 receptor partial agonist, in common marmosets. Psychopharmacology (Berl). 2015 Sep;232(17):3287-95.
[2]. Powell SB, et al. RO-10-5824 is a selective dopamine D4 receptor agonist that increases novel object exploration in C57 mice. Neuropharmacology. 2003 Mar;44(4):473-81 |
| Additional Infomation |
1. Ro 10-5824 dihydrochloride is a synthetic small-molecule selective dopamine D4 receptor (D4R) partial agonist initially developed for preclinical studies of D4R-mediated cognitive and neurophysiological functions [1][2]. 2. Mechanism of action: This compound binds to the D4R binding site and acts as a partial agonist, modulating G protein (Gi/o)-mediated cAMP inhibition and β-arrestin-dependent signaling, preferentially activating the prefrontal cortex D4R circuit, thereby enhancing attention, reducing impulsivity, and promoting exploration of new objects [1][2]. 3. Research applications: It has been widely used as a D4R-selective tool compound in preclinical studies to elucidate the role of D4R in cognitive function (attention/working memory), neurophysiology (prefrontal cortex neuronal firing), and mental illness models (schizophrenia/attention deficit hyperactivity disorder). 4. Selectivity advantage: Its high selectivity for D4R (relative to other dopamine receptors and non-targeted GPCRs) eliminates the confounding effects of D1/D2 receptor activation, making it the gold standard tool for studying the specific mechanisms of D4R [2]
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| Molecular Formula |
C17H22CL2N4
|
|---|---|
| Molecular Weight |
353.289381504059
|
| Exact Mass |
352.122
|
| CAS # |
189744-94-3
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| PubChem CID |
16759174
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| Appearance |
Typically exists as solid at room temperature
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
23
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| Complexity |
362
|
| Defined Atom Stereocenter Count |
0
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| SMILES |
NC1=NC(C)=NC=C1CN2CC=C(C3=CC=CC=C3)CC2.[H]Cl.[H]Cl
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| InChi Key |
UZHMRJRDYCRKIZ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C17H20N4.2ClH/c1-13-19-11-16(17(18)20-13)12-21-9-7-15(8-10-21)14-5-3-2-4-6-14;;/h2-7,11H,8-10,12H2,1H3,(H2,18,19,20);2*1H
|
| Chemical Name |
2-methyl-5-[(4-phenyl-3,6-dihydro-2H-pyridin-1-yl)methyl]pyrimidin-4-amine;dihydrochloride
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
H2O : ~100 mg/mL (~283.05 mM)
DMSO : ~8.33 mg/mL (~23.58 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 50 mg/mL (141.53 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8305 mL | 14.1527 mL | 28.3054 mL | |
| 5 mM | 0.5661 mL | 2.8305 mL | 5.6611 mL | |
| 10 mM | 0.2831 mL | 1.4153 mL | 2.8305 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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