| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| Other Sizes |
Purity: ≥98%
PF-05180999 is a phosphodiesterase 2A (PDE2A) inhibitor, with an IC50 of 1.6 nM. PF-05180999 was identified as a preclinical candidate targeting cognitive impairment associated with schizophrenia. PF-05180999 demonstrated potent binding to PDE2A in brain tissue, dose responsive mouse brain cGMP increases, and reversal of N-methyl-d-aspartate (NMDA) antagonist-induced (MK-801, ketamine) effects in electrophysiology and working memory models in rats. Preclinical pharmacokinetics revealed unbound brain/unbound plasma levels approaching unity and good oral bioavailability resulting in an average concentration at steady state (Cav,ss) predicted human dose of 30 mg once daily (q.d.). Modeling of a modified release formulation suggested that 25 mg twice daily (b.i.d.) could maintain plasma levels of PF-05180999 at or above targeted efficacious plasma levels for 24 h, which became part of the human clinical plan.
| Targets |
Phosphodiesterase 2A (PDE2A) (IC50 = 1.6 nM)
Highly selective over other PDE families (PDE1B1, 3A1, 4D3, 5A1, 6, 7B, 8B, 9A1, 10A1, 11A4). Displaces radioligand 2 from native PDE2A in rat, dog, and monkey striatal tissue with Ki values of 4.2 nM, 8.4 nM, and 5.5 nM, respectively. [1] |
|---|---|
| ln Vitro |
The phosphodiesterase 2A (PDE2A) inhibitor PF-05180999 has an IC50 of 1.6 nM. With Kis of 4.2, 8.4, and 5.5 nM and IC50 of 2.6, 5.2, and 3.4 nM, respectively, PF-05180999 binds to PDE2A in rats, dogs, and monkeys. With an IC50 of 2.03 μM (PDE10A1), 26.969 μM (PDE7B), 50.09 μM (PDE11A4), and >56.25 μM (PDE1B1, PDE3A1, PDE4D3, PDE5A1, PDE6 (bovine), PDE8B, and PDE9A1), respectively, PF-05180999 shows limited activity against PDEs. In addition to not directly inhibiting human recombinant cytochrome P450 (CYP) enzymes (1A2, 2B6, 2C8, 2C9, 2C19, 2D6, and 3A), PF-05180999 is a modest inducer of CYP3A4 and does not stimulate CYP1A2 [1].
In human liver microsomes (HLM), PF-05180999 shows an intrinsic clearance (Clint) of 52 mL/min/kg. [1] Exhibits low efflux ratio (MDR BA/AB = 1.4) in MDCK-MDR1 cells, indicating good brain penetration potential. [1] Shows high passive permeability in RRCK cells (26 x 10⁻⁶ cm/s). [1] Metabolite identification studies in HLM identified oxidation at the azetidine ring, yielding an acid metabolite and a 3-hydroxy azetidine metabolite, both significantly less active. [1] Demonstrated minimal activity across a panel of kinases and other targets, confirming high selectivity for PDE2A. [1] No direct or time-dependent inhibition of major human CYP enzymes (1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 3A) was observed. It was identified as a weak inducer of CYP3A4, but the risk for drug-drug interactions at clinically relevant exposures was predicted to be low. Primary metabolism is via CYP3A4. [1] |
| ln Vivo |
PF-05180999 (Compound 30; 0.032-0.32 mg/kg mg/kg, subcutaneous injection) significantly lowers ketamine-produced working memory mistakes in the rat radial arm maze (RAM) model of working memory. PF-05180999 induces an acute and exposure-dependent rise in the accumulation of global levels of cGMP in the cortex, striatum, and hippocampus, but without changes in cAMP and the related downstream phosphorylated cAMP response element binding protein (p-CREB). mice[1].
Administration to mice led to an acute, exposure-dependent increase in cGMP levels in cortex, striatum, and hippocampus, but no changes in cAMP or p-CREB. The relationship between unbound brain concentration (Cbu) and cGMP increase was linear, with Cbu E50 values estimated at 79.4 nM (cortex), 98.0 nM (striatum), and 116.3 nM (hippocampus). [1] In rats, it reversed the disruption of cortical low-frequency delta oscillations induced by the NMDA antagonist MK-801 (0.07 mg/kg, IV) in an EEG model. The Cbu associated with 80% effect reversal (IC80) was 9 nM. [1] In a rat radial arm maze working memory model disrupted by ketamine, PF-05180999 (0.032 to 0.32 mg/kg, SC) significantly attenuated the errors, with efficacious Cbu ranging from 3.1 to 12.5 nM. A U-shaped dose-response was observed, with attenuation of effect at 1 mg/kg. [1] No effects on spontaneous or habituated locomotor activity in rats (SC, 0.32-3.2 mg/kg). [1] No effects in the conditioned avoidance responding assay, and did not alter the efficacy of risperidone when co-administered. [1 A positron emission tomography study in non-human primates showed 50% PDE2A occupancy at Cbu = 57 nM. [1] |
| Enzyme Assay |
The PDE2A enzyme inhibition assay was performed as a scintillation proximity assay (SPA). Human PDE2A3 enzyme was used. Test compounds were diluted in DMSO/water and tested in triplicate across 8 concentrations. The reaction mixture contained the enzyme, the substrate [³H]cGMP at a sub-Km concentration, and the test compound in assay buffer. After incubation at room temperature for a determined period (time set for 10-20% substrate hydrolysis), the reaction was terminated by adding SPA beads that preferentially bind the hydrolyzed product, [³H]GMP. Radioactivity was measured using a scintillation counter. IC50 values were determined from concentration-effect curves using non-linear regression. [1]
PDE selectivity assays for other PDE families followed a similar SPA format. For cAMP-hydrolyzing PDEs, [³H]cAMP was used as substrate; for cGMP-hydrolyzing PDEs, [³H]cGMP was used. The assay buffer composition varied slightly (e.g., PDE1B1 assay included CaCl₂ and calmodulin). The reaction was started by adding the radiolabeled nucleotide, incubated, stopped with SPA beads, and counted. [1] |
| Cell Assay |
Metabolite identification studies were conducted in liver microsomes or hepatocytes from rat, dog, and human. Microsomal/hepatocyte suspensions were incubated with PF-05180999 (1-50 µM) in phosphate buffer with an NADPH-regenerating system at 37°C for 1 hour. Reactions were quenched with acetonitrile, centrifuged, and the supernatant was evaporated. The residue was reconstituted and analyzed by LC-MS/MS to identify metabolites. [1]
The MDR BA/AB efflux ratio was determined using Madin-Darby Canine Kidney cells transfected with the human MDR1 gene. The ratio of basolateral-to-apical flux to apical-to-basolateral flux was measured to assess P-glycoprotein-mediated efflux. [1] |
| Animal Protocol |
Mouse brain cGMP study: CD-1 mice were administered PF-05180999 subcutaneously (SC) as a solution in 20% sulfobutylether-β-cyclodextrin acidified with HCl. At designated times, animals were euthanized by focused microwave irradiation. Brain regions (cortex, striatum, hippocampus) were dissected, homogenized in HCl, and centrifuged. cGMP levels in the supernatant were quantified using an enzyme immunoassay kit. Satellite groups were used for pharmacokinetic exposure assessment. [1]
Rat cortical EEG study: Male Sprague-Dawley rats under urethane anesthesia were placed in a stereotaxic frame. EEG electrodes were implanted in the medial prefrontal cortex. The NMDA antagonist MK-801 (0.07 mg/kg) was administered intravenously (IV) to disrupt delta oscillations. PF-05180999 was administered IV, and EEG signals were recorded and analyzed for power in the low-frequency delta band (0.5-1.8 Hz). [1] Rat radial arm maze study: Male Long-Evans rats were food-restricted and trained on an 8-arm radial maze task. After meeting a performance criterion, they were tested. On test days, PF-05180999 (or vehicle) was administered SC, followed by ketamine (or saline). The number of working memory errors (re-entries into previously visited arms) was recorded. [1] Neuropharmacokinetic studies: PF-05180999 was administered to mice (SC), rats (IV/SC), dogs (oral), and monkeys (SC) via various formulations (e.g., solution in cyclodextrin, suspension in methylcellulose). Blood (for plasma), cerebrospinal fluid (CSF) via cisterna magna puncture, and whole brain were collected at designated time points. Tissues were processed (brain homogenized), and all samples were stored at -80°C before bioanalysis. [1] Toxicology studies: Compounds were evaluated in rat oral exploratory toxicity studies for up to 15 days at doses of 5, 50, and 250 mg/kg/day. Subsequent single and repeat-dose studies (up to 1 month) were conducted in rats and dogs with PF-05180999. Animals were monitored for clinical signs, and tissues were examined histopathologically. [1] |
| ADME/Pharmacokinetics |
PF-05180999 showed stable brain permeability in different species, with an average brain tissue to plasma free drug concentration ratio (Cbu/Cpu) of 0.65 and an average cerebrospinal fluid to plasma free drug concentration ratio (CSF/Cpu) of 0.98. [1]
In rats, after intravenous injection of 1 mg/kg, the plasma clearance rate (CLp) was high (122 mL/min/kg), the steady state distribution volume (Vss) was 3.9 L/kg, and the terminal half-life (t1/2) was 1.4 hours. The oral bioavailability (F%) was 44% at a dose of 5 mg/kg. [1] In dogs, after intravenous injection of 0.3 mg/kg, the CLp was low (12.3 mL/min/kg), the Vss was 5.0 L/kg, and the t1/2 was 4.1 hours. The oral bioavailability was 57% at 5 mg/kg. [1] The predicted human pharmacokinetic parameters were: CLp = 8.8 mL/min/kg, Vss = 1.9 L/kg, t1/2 = 2.5 hours, and bioavailability = 32%. [1] The mean steady-state plasma concentration (Cav,ss) to achieve an effective therapeutic effect was estimated to be 11 ng/mL. The predicted human dose for an immediate-release formulation to achieve this Cav,ss was 30 mg once daily (QD). [1] The model indicated that a sustained-release formulation of 25 mg twice daily (BID) could maintain plasma concentrations above the effective threshold for 24 hours. [1] |
| Toxicity/Toxicokinetics |
In a 15-day exploratory toxicity study in rats, PF-05180999 was generally well tolerated at doses up to 250 mg/kg/day, and no evidence of cardiac or vascular damage was observed despite the high exposure. [1]
In subsequent 1-month studies in rats and dogs, the target organs of PF-05180999 were identified as skeletal muscle (rat) and central nervous system (rat and dog). No effects on the heart or blood vessels were observed. [1] In a single-dose canine study, the tolerated dose of PF-05180999 was up to 30 mg/kg. In a 14-day canine study, no microscopic lesions were observed at doses up to 15 mg/kg/day. [1] The human plasma protein binding rate (fractional free protein, fup) was 0.341. [1] |
| References | |
| Additional Infomation |
PF-05180999 is currently undergoing clinical trial NCT01530529 (a study evaluating the relative bioavailability of a sustained-release formulation of PF-05180999). PF-05180999 has been identified as a preclinical candidate for schizophrenia-associated cognitive impairment (CIAS). [1] Its mechanism of action involves potent and selective inhibition of PDE2A, thereby increasing cGMP levels in key cognitive regions of the brain (frontal cortex, striatum, hippocampus). In electrophysiological and working memory models, it has been able to reverse cognitive deficits induced by NMDA receptor antagonists (MK-801, ketamine), suggesting its potential to treat insufficient glutamatergic signaling in CIAS. [1] A sustained-release (MR) formulation has been developed to maintain stable plasma concentrations, and a 25 mg twice-daily (BID) sustained-release formulation is planned for evaluation in human clinical trials. [1]
Due to the improved lipophilic ligand efficiency (LipE), the expected human dose is significantly lower compared to the initial lead compound 1 (108 mg/day). [1] |
| Molecular Formula |
C19H17F3N8
|
|---|---|
| Molecular Weight |
414.387092351913
|
| Exact Mass |
414.152
|
| CAS # |
1394033-54-5
|
| PubChem CID |
60143346
|
| Appearance |
White to yellow solid powder
|
| Density |
1.6±0.1 g/cm3
|
| Index of Refraction |
1.720
|
| LogP |
0.28
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
9
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
30
|
| Complexity |
617
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
CLGCHUKGBICQTE-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C19H17F3N8/c1-11-27-15(17-18(29-6-3-7-29)24-10-26-30(11)17)13-9-25-28(2)16(13)14-5-4-12(8-23-14)19(20,21)22/h4-5,8-10H,3,6-7H2,1-2H3
|
| Chemical Name |
4-(1-Azetidinyl)-7-methyl-5-[1-methyl-5-[5-(trifluoromethyl)-2-pyridinyl]-1H-pyrazol-4-yl]-imidazo[5,1-f][1,2,4]triazine
|
| Synonyms |
PF 5180999; PF999; PF-999; PF 999; PF05180999; PF-05180999; PF 5180999; PF5180999; PF-5180999;
|
| HS Tariff Code |
2934.99.9001
|
| 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 : ~50 mg/mL (~120.66 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.03 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 25.0 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: ≥ 2.5 mg/mL (6.03 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (6.03 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4132 mL | 12.0659 mL | 24.1319 mL | |
| 5 mM | 0.4826 mL | 2.4132 mL | 4.8264 mL | |
| 10 mM | 0.2413 mL | 1.2066 mL | 2.4132 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA