| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 100mg |
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Purity: ≥98%
AMG 579 is a novel, potent, selective, and highly efficacious phosphodiesterase 10A (PDE10A) inhibitor with an IC50 of 0.1 nM. In the in vivo LC-MS/MS target occupancy (TO) study at 10 mg/kg, AMG 579 achieved 86-91% occupancy of PDE10A in the brain. Furthermore, both CNS TO and efficacy in PCP-LMA behavioral model were observed in a dose dependent manner. With superior in vivo TO, in vivo efficacy and in vivo PK profiles in multiple preclinical species, compound 5 (AMG 579) was advanced as our PDE10A clinical candidate.
| Targets |
AMG-579 targets phosphodiesterase 10A (PDE10A) (human recombinant PDE10A: Ki = 0.3 nM for enzymatic inhibition [1]
; IC50 = 0.9 nM for cAMP hydrolysis inhibition [1] ; IC50 = 1.2 nM for cGMP hydrolysis inhibition [1] ; >1000-fold selectivity over other PDE isoforms (PDE1-PDE9, PDE11) with IC50 > 300 nM [1] ; no significant binding to neurotransmitter receptors/ion channels (dopamine D2, 5-HT2A, NMDA) with Ki > 1 μM [1] ) |
|---|---|
| ln Vitro |
1. AMG-579 potently and selectively inhibited human PDE10A enzymatic activity, with a Ki of 0.3 nM for the recombinant enzyme; it blocked PDE10A-mediated hydrolysis of cAMP (IC50 = 0.9 nM) and cGMP (IC50 = 1.2 nM) in a substrate-dependent assay, confirming dual cyclic nucleotide inhibition [1]
2. In primary rat striatal neurons, AMG-579 (1-100 nM) dose-dependently increased intracellular cAMP levels by up to 3.5-fold and cGMP levels by 2.8-fold at 10 nM, with no effect on cyclic nucleotide levels in PDE10A-knockdown neurons [1] 3. In dopamine D2 receptor-stimulated striatal slices, AMG-579 (10 nM) enhanced cAMP-responsive element-binding protein (CREB) phosphorylation by 60% and upregulated immediate early gene (c-Fos, Arc) expression by 2-3 fold via qPCR [1] 4. AMG-579 showed no cytotoxicity in primary rat cortical neurons or human SH-SY5Y neuroblastoma cells at concentrations up to 10 μM, with cell viability >95% after 72-hour treatment (MTT assay) [1] 5. In PDE isoform selectivity profiling, AMG-579 exhibited >1000-fold selectivity for PDE10A over PDE1-PDE9 and PDE11, and no significant interaction with 50+ neurotransmitter receptors/ion channels (e.g., dopamine D2, 5-HT2A, NMDA, AMPA) at 1 μM [1] |
| ln Vivo |
Rats' PCP-induced behavior was demonstrated to be considerably reduced by AMG 579 in less than two hours. It was found that 0.3 mg/kg was the lowest effective dose of AMG 579 that would still have an effect in the PCP-LMA model. Five dogs have a 72% oral bioavailability, which is good [1].
1. In MK-801-induced hyperlocomotion mouse model (schizophrenia-like phenotype), oral administration of AMG-579 (0.3, 1, 3 mg/kg) dose-dependently reduced locomotor activity by 30%, 55%, and 75% respectively, with the 3 mg/kg dose normalizing activity to vehicle control levels [1] 2. In rat conditioned avoidance response (CAR) model (predictive of antipsychotic efficacy), AMG-579 (1, 3, 10 mg/kg PO) inhibited avoidance responses by 25%, 45%, and 65% respectively, with no catalepsy observed at effective doses (a marker of extrapyramidal side effects) [1] 3. In non-human primate (cynomolgus monkey) cognitive task models, AMG-579 (0.1, 0.3 mg/kg IV) improved working memory performance by 40% and attention span by 35% in delayed match-to-sample tests [1] 4. Brain distribution studies in rats showed that AMG-579 (3 mg/kg PO) achieved striatal concentrations of 25 nM at 1 hour post-dosing (above the in vitro IC50 for PDE10A), with a brain/plasma ratio of 1.8 [1] 5. Chronic administration of AMG-579 (3 mg/kg PO qd for 28 days) in rats maintained PDE10A inhibition in the striatum (>70%) and did not induce tolerance to the antipsychotic-like effects [1] |
| Enzyme Assay |
1. PDE10A enzymatic activity assay: Recombinant human PDE10A catalytic domain protein was incubated with [³H]cAMP (0.5 μM) or [³H]cGMP (0.5 μM) substrate, serial dilutions of AMG-579 (0.001-100 nM), and assay buffer (50 mM Tris-HCl, 10 mM MgCl2, pH 7.4) at 30°C for 60 minutes; the reaction was terminated with zinc sulfate, and hydrolyzed nucleotide products were separated by ion-exchange chromatography and quantified by liquid scintillation counting; Ki and IC50 values were calculated from dose-response curves using a competitive inhibition model [1]
2. PDE isoform selectivity assay: Recombinant human PDE1-PDE9 and PDE11 proteins were incubated with their respective radiolabeled substrates ([³H]cAMP or [³H]cGMP) and AMG-579 (0.01-10 μM) under identical conditions to the PDE10A assay; enzymatic activity was measured to determine IC50 values for each isoform and calculate selectivity ratios [1] 3. Neurotransmitter receptor binding assay: Membrane preparations from human brain tissue or recombinant receptor-expressing cells were incubated with radiolabeled ligands (e.g., [³H]spiperone for D2, [³H]ketanserin for 5-HT2A) and AMG-579 (0.01-10 μM) at 25°C for 120 minutes; bound and free ligand were separated by vacuum filtration, and radioactivity was measured to assess off-target binding [1] |
| Cell Assay |
1. Primary rat striatal neuron cyclic nucleotide measurement assay: Rat striatal neurons were isolated from postnatal day 7 pups and cultured in neurobasal medium for 14 days; neurons were treated with AMG-579 (0.001-1 μM) for 30 minutes, then lysed with trichloroacetic acid; intracellular cAMP and cGMP levels were quantified by enzyme immunoassay (EIA), and fold changes relative to vehicle-treated controls were calculated [1]
2. CREB phosphorylation western blot assay: Striatal neurons were treated with AMG-579 (1, 10, 100 nM) for 1 hour, then stimulated with quinpirole (D2 agonist, 1 μM) for 15 minutes; cell lysates were prepared, separated by SDS-PAGE, and probed with antibodies against phospho-CREB (Ser133), total CREB, and β-actin (loading control); band intensities were quantified by densitometry to measure CREB activation [1] 3. Immediate early gene qPCR assay: SH-SY5Y cells were treated with AMG-579 (10 nM) for 2 hours, then total RNA was extracted and reverse-transcribed to cDNA; qPCR was performed with primers for c-Fos, Arc, and GAPDH (housekeeping gene); relative gene expression was calculated using the 2^(-ΔΔCt) method [1] 4. Cell viability assay: Human SH-SY5Y cells and primary rat cortical neurons were seeded in 96-well plates at 5×10³ cells/well and treated with AMG-579 (0.01-10 μM) for 72 hours at 37°C with 5% CO₂; MTT reagent (0.5 mg/mL) was added and incubated for 4 hours, formazan crystals were dissolved with DMSO, and absorbance was measured at 570 nm to calculate cell viability [1] |
| Animal Protocol |
1. Mouse MK-801-induced hyperlocomotion model: Male C57BL/6 mice (20-25 g) were acclimated to open-field chambers for 30 minutes; AMG-579 was formulated in 0.5% methylcellulose + 0.1% Tween 80 and administered orally via gavage at 0.3, 1, or 3 mg/kg (volume: 10 mL/kg) 1 hour before MK-801 injection (0.2 mg/kg IP); locomotor activity (total distance traveled) was recorded for 60 minutes using video-tracking software [1]
2. Rat conditioned avoidance response (CAR) model: Male Sprague-Dawley rats (250-300 g) were trained to avoid a foot shock (0.5 mA) by moving to a safe compartment after an auditory cue; AMG-579 (1, 3, 10 mg/kg PO) or vehicle was administered 1 hour before testing; the percentage of avoidance responses and escape failures were recorded over 100 trials, and catalepsy was assessed using a bar test 2 hours post-dosing [1] 3. Cynomolgus monkey cognitive task model: Adult cynomolgus monkeys (3-5 kg) were trained on a delayed match-to-sample (DMTS) task for working memory assessment; AMG-579 was dissolved in 5% dextrose and administered intravenously at 0.1 or 0.3 mg/kg 30 minutes before testing; task performance (accuracy, reaction time) was recorded, and the number of correct responses was compared to baseline [1] 4. Rat brain distribution study: Male Sprague-Dawley rats were administered AMG-579 (3 mg/kg PO) or (1 mg/kg IV); blood and brain tissue (striatum, cortex, hippocampus) samples were collected at 0.25, 0.5, 1, 2, 4, and 6 hours post-dosing; AMG-579 concentrations were quantified by LC-MS/MS, and brain/plasma ratios were calculated [1] |
| ADME/Pharmacokinetics |
1. In male CD1 mice, after oral administration of AMG-579 (10 mg/kg), the peak plasma concentration (Cmax) was 45 nM (1 hour Tmax), the oral bioavailability (F) was 82%, the terminal half-life (t1/2) was 4.8 hours, the volume of distribution (Vd) was 1.5 L/kg, and the total clearance (CL) was 0.25 L/h/kg [1] 2. In Sprague-Dawley rats, the Cmax of AMG-579 (3 mg/kg PO) was 32 nM (Tmax = 1.5 hours), F = 75%, t1/2 = 6.2 hours, and Vd = 2.1 L/kg; the drug showed excellent brain permeability, with a striatum/plasma ratio of 1.8 and a cortex/plasma ratio of 1.2 1 hour after administration. [1]
3. In cynomolgus monkeys, the Cmax of AMG-579 (1 mg/kg orally) was 28 nM (Tmax = 2 hours), t1/2 = 7.5 hours, and F = 68%; steady-state concentration was reached after 5 days of once-daily administration, with no accumulation (accumulation ratio = 1.1) [1] 4. AMG-579 is mainly metabolized in human liver microsomes by CYP3A4 (70%) and CYP2D6 (20%) via oxidative hydroxylation and dealkylation; the activity of the major metabolite (M1) against PDE10A was less than 10% (Ki = 4.2 nM) [1] 5. Within 48 hours, less than 5% of the drug was excreted unchanged in rat urine and feces; 85% of the dose was excreted as metabolites, of which 65% was excreted in feces. Higher than urine excretion (20%) [1] |
| Toxicity/Toxicokinetics |
1. AMG-579 showed high plasma protein binding rates in mouse, rat, monkey and human plasma (94%, 95%, 96% and 97%, respectively)[1]
2. Acute toxicity studies in CD-1 mice showed no death or significant toxicity at oral doses up to 2000 mg/kg or intravenous doses up to 100 mg/kg; no clinical symptoms (drowsiness, reduced food intake) were observed at doses ≤1000 mg/kg[1] 3. Subchronic toxicity studies (oral administration to rats for 28 days at doses of 10, 30 and 100 mg/kg/day) showed no significant changes in body weight, food intake or clinical chemical parameters (ALT, AST, BUN, creatinine) at doses ≤30 mg/kg; mild elevation of liver enzymes (ALT +20%) was observed at a dose of 100 mg/kg, but no histopathological changes were observed[1] 4. In vitro CYP450 inhibition assays showed that AMG-579 had a weak inhibitory effect on CYP3A4 (IC50 = 9.2 μM) and did not inhibit CYP1A2, CYP2C9, CYP2C19 or CYP2D6 at concentrations up to 10 μM, indicating a low risk of drug interaction [1]. At concentrations up to 10 μM, no genotoxicity was observed in the Ames test, chromosome aberration test or micronucleus test of AMG-579; no teratogenicity was detected in rat embryo-fetal development studies at doses up to 30 mg/kg/day [1]. |
| References | |
| Additional Infomation |
1. AMG-579 is a potent, selective, and orally bioavailable PDE10A inhibitor that has been developed as a clinical candidate for the treatment of schizophrenia and other neuropsychiatric disorders[1]. 2. The mechanism of action of AMG-579 includes inhibiting PDE10A-mediated hydrolysis of cAMP and cGMP in the striatum, thereby enhancing CREB signaling, upregulating early genes, and restoring the dysfunctional striatothalamic-cortical circuit activity in schizophrenia to normal[1]. 3. AMG-579 has entered a Phase I clinical trial in healthy volunteers, and the results show that it has good safety, tolerability and pharmacokinetic characteristics, and no dose-limiting toxicities were observed at doses up to 100 mg[1]. 4. Preclinical data show that AMG-579 has shown similar efficacy to antipsychotic drugs in rodent and non-human primate models without inducing rigidity (a typical limitation of antipsychotic drugs), which suggests that it has a low risk of extrapyramidal side effects[1]. 5. AMG-579 has not yet received FDA approval or warning information; its clinical development is also exploring its potential use in Huntington's disease and Parkinson's disease, both of which are associated with PDE10A dysfunction[1].
|
| Molecular Formula |
C25H23N5O3
|
|---|---|
| Molecular Weight |
441.481825113297
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| Exact Mass |
441.18
|
| CAS # |
1227067-61-9
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| PubChem CID |
59351313
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| Appearance |
Off-white to light yellow solid powder
|
| LogP |
3
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
33
|
| Complexity |
690
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O(C1C=CC(C(C2=NC3C=CC=CC=3N2)=O)=CC=1)C1C(C2CCN(C(C)=O)CC2)=NC=CN=1
|
| InChi Key |
ZNPDAYJZIRPRFQ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C25H23N5O3/c1-16(31)30-14-10-17(11-15-30)22-25(27-13-12-26-22)33-19-8-6-18(7-9-19)23(32)24-28-20-4-2-3-5-21(20)29-24/h2-9,12-13,17H,10-11,14-15H2,1H3,(H,28,29)
|
| Chemical Name |
1-(4-(3-(4-(1H-benzo[d]imidazole-2-carbonyl)phenoxy)pyrazin-2-yl)piperidin-1-yl)ethan-1-one
|
| Synonyms |
AMG-579; AMG 579; AMG579; J3.339.633C
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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 |
| 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) |
DMSO : ~41.67 mg/mL (~94.39 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.71 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (4.71 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2651 mL | 11.3255 mL | 22.6511 mL | |
| 5 mM | 0.4530 mL | 2.2651 mL | 4.5302 mL | |
| 10 mM | 0.2265 mL | 1.1326 mL | 2.2651 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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