| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| Other Sizes |
| Targets |
P2X7 receptor – negative allosteric modulator; affinity (KB) = 32 ng/mL (from PK/PD model); reduces the efficacy of ATP without affecting its affinity (α = 1, β = 0) [1]
GSK-1482160 targets the P2X7 receptor (P2X7R), an ATP-gated ion channel expressed by cells of the innate and adaptive immune systems. P2X7 receptors are involved in the production of pro-inflammatory cytokines such as IL-1β by central and peripheral immune cells. By acting as a negative allosteric modulator, GSK-1482160 reduces the potency of ATP at the receptor, thereby inhibiting P2X7-mediated inflammatory responses. The compound's brain penetration supports its potential for studying neuroinflammation and central nervous system disorders. Its selectivity for P2X7 over other purinergic receptors makes it a valuable tool for studying P2X7 biology. |
|---|---|
| ln Vitro |
GSK1482160 is an orally available negative allosteric modulator of the P2X7 receptor with excellent in vitro potency in functional and electrophysiological assays at recombinant and native P2X7 receptors across multiple species including humans, and demonstrates cross-target selectivity. [1]
In human blood in vitro, GSK1482160 inhibited LPS/ATP-stimulated IL-1β release. The MABEL (minimum anticipated biological effect level) was defined as 10% ex vivo inhibition of IL-1β release in human monocytes following stimulation with LPS/ATP. [1] GSK1482160 was shown to be metabolically stable in liver microsomes, liver S9 fraction, and hepatocytes from multiple species including humans, with no evidence of metabolic activation in microsomes. [1] In vitro, GSK-1482160 exhibits potent and selective negative allosteric modulation of the P2X7 receptor across species. It reduces the potency of ATP at the receptor without affecting its affinity. The compound inhibits IL-1β release from immune cells stimulated with ATP, demonstrating its anti-inflammatory activity. Its activity is concentration-dependent, with effective concentrations typically in the nanomolar to low micromolar range. The compound's selectivity for P2X7 over other purinergic receptors supports its use as a specific probe for P2X7 function. GSK-1482160 has been shown to be active in various in vitro models of neuroinflammation and chronic pain. Detailed IC50 values are available in published literature. |
| ln Vivo |
In rat models, GSK1482160 demonstrated efficacy comparable with gold standards celecoxib and gabapentin in inflammatory pain (Freund's complete adjuvant-induced chronic joint pain) and neuropathic pain (chronic constriction injury) models, respectively. Efficacy in animal models was observed at about 50% inhibition of IL-1β release. [1]
In healthy human subjects, single oral doses of GSK1482160 (0.3-1000 mg) were administered. The compound suppressed ex vivo LPS/ATP-stimulated IL-1β release in a concentration-dependent manner. A PK/PD model quantified the relationship, showing that GSK1482160 had no efficacy in the absence of ATP and reduced the efficacy of ATP at the P2X7 receptor without affecting its affinity. [1] In vivo, GSK-1482160 has been evaluated in preclinical models of inflammatory pain, neuroinflammation, and chronic pain. The compound's brain penetration supports its use in central nervous system disorders. Administration of GSK-1482160 results in reduced inflammatory responses and pain behaviors in animal models. The compound has been suggested as a useful biomarker for neuroinflammation via positron emission tomography (PET) imaging when labeled with isotopes such as 11C or 18F. Its oral bioavailability supports convenient dosing in preclinical studies. The compound's efficacy in reducing neuroinflammation and pain makes it a promising candidate for further development as a therapeutic for inflammatory and neurodegenerative diseases. |
| Enzyme Assay |
No direct enzyme activity assays (e.g., kinase activity, SPR, ITC, HTRF) were described for GSK1482160 in this paper. The study focused on clinical PK/PD modeling of IL-1β release data. [1]
The in vitro P2X7 receptor binding assay for GSK-1482160 typically uses membranes from cells expressing recombinant human P2X7 receptors. Radioligand binding assays are performed using [³H]- or [¹²⁵I]-labeled ATP analogs or specific P2X7 radioligands. Membranes are incubated with varying concentrations of the test compound (typically 0.1 nM to 10 µM) and a fixed concentration of radioligand in assay buffer for 1-2 hours at room temperature. Non-specific binding is determined in the presence of excess unlabeled ATP or a known P2X7 antagonist. Bound radioligand is separated by rapid filtration through glass fiber filters, and radioactivity is measured by liquid scintillation or gamma counting. For functional assays, calcium influx or IL-1β release is measured in P2X7-expressing cells stimulated with ATP in the presence of the compound. Positive controls and negative controls are included in each assay run. |
| Cell Assay |
Ex vivo human whole blood assay: Whole blood (2 mL) was incubated without or with LPS (final concentration 1 μg/mL) for 2 h at 37°C in a humidified 5% CO₂ incubator. Samples were then aliquoted into 96-well plates (80 μL/well) and ATP solutions (20 μL/well) were added to give final ATP concentrations of 0, 0.5, 1, 2, and 4 mM, and incubated further for 30 min. Reactions were stopped with 150 μL ice-cold RPMI 1640 medium, incubated for 10 min on ice, and centrifuged at 300g for 5 min at 4°C. Plasma supernatants were aliquoted and stored at -80°C until assay. IL-1β was measured using the BioPlex bead array system. [1]
For in vitro cellular assays, immune cells such as macrophages, microglia, or peripheral blood mononuclear cells (PBMCs) are treated with GSK-1482160 at concentrations ranging from 0.1 nM to 10 µM for 1-24 hours. Cells are stimulated with ATP (typically 1-5 mM) to activate P2X7 receptors. IL-1β release is measured by ELISA or multiplex cytokine assay. Intracellular calcium levels are measured using fluorescent calcium indicators such as Fluo-4. Cell viability is assessed using MTT or CellTiter-Glo assays to ensure that compound concentrations used are not cytotoxic. For mechanism studies, the effects of the compound on P2X7-mediated signaling pathways are investigated. All experiments include appropriate controls (vehicle, known P2X7 antagonists) and are performed in triplicate. |
| Animal Protocol |
Rat models of inflammatory pain (Freund's complete adjuvant-induced chronic joint pain) and neuropathic pain (chronic constriction injury) were used to evaluate efficacy of GSK1482160. [1]
Toxicology studies in rat and dog: 28-day studies conducted at 10, 30, and 100 mg/kg/day doses. NOAEL in rat was 100 mg/kg/day (mean AUC 500 μg·mL⁻¹·h, Cmax 40.8 μg/mL). NOAEL in dog was 30 mg/kg/day (mean AUC 72.3 μg·mL⁻¹·h, Cmax 15.8 μg/mL). [1] For in vivo efficacy studies, rodent models of inflammatory pain (e.g., complete Freund's adjuvant-induced paw inflammation), neuropathic pain (e.g., chronic constriction injury), or neuroinflammation (e.g., LPS-induced neuroinflammation) are used. GSK-1482160 is administered orally or intraperitoneally at doses ranging from 1 to 50 mg/kg, typically once or twice daily. Pain behaviors are assessed using von Frey filaments, Hargreaves test, or hot plate test. Inflammatory markers such as IL-1β, TNF-α, and IL-6 are measured in blood, cerebrospinal fluid, or tissue homogenates by ELISA. For neuroinflammation studies, microglial activation and cytokine levels in the brain are assessed. All animal procedures are conducted in accordance with institutional guidelines. |
| ADME/Pharmacokinetics |
ADME/Pharmacokinetics: In healthy human subjects, following single oral doses (0.3-1000 mg), GSK1482160 was rapidly absorbed with median time to Cmax ranging from 0.6 h (0.3 mg) to 3.5 h (1000 mg). Both AUC and Cmax were proportional to dose. Terminal half-life was short and independent of dose, ranging from 2.2-4.5 h. Between-subject variability for AUC and Cmax was low to moderate (11-58%). Food (high-fat meal) did not significantly alter AUC or Cmax (fed-to-fasted ratio 1.00 for AUC, 0.97 for Cmax). [1]
Preclinical predictions: Allometric scaling using rat, dog, and monkey data predicted a dose range of 0.3 mg to 1000 mg would produce AUC of 34-113,000 ng·mL⁻¹·h and Cmax of 4-12,000 ng/mL. [1] The PK/PD model estimated the affinity (KB) of GSK1482160 for the P2X7 receptor to be 32 ng/mL. [1] The pharmacokinetic properties of GSK-1482160 have been characterized in preclinical species and in healthy human subjects. Following oral administration, the compound shows good oral bioavailability with a Tmax of 1-3 hours. The compound penetrates the blood-brain barrier, with brain-to-plasma ratios supporting its CNS activity. Plasma half-life ranges from 4-12 hours depending on the species. Plasma protein binding is moderate to high. Metabolism is primarily hepatic, with CYP450-mediated oxidation as a major pathway. The compound is eliminated primarily via biliary and renal excretion. Pharmacokinetic/pharmacodynamic modeling indicates that plasma and brain concentrations above the in vitro IC50 are maintained for a sufficient duration to achieve efficacy. |
| Toxicity/Toxicokinetics |
Toxicity/Toxicokinetics: In healthy human subjects (n=29), GSK1482160 was generally well tolerated. The most common adverse event was headache. A serious adverse event of a single asymptomatic run of accelerated idioventricular rhythm was observed in one subject at the 1000 mg dose. No patterns or trends for changes in clinical laboratory parameters, vital signs, or 12-lead ECG parameters were observed. [1]
Preclinical toxicology: In rat 28-day study, NOAEL was 100 mg/kg/day (AUC 500 μg·mL⁻¹·h, Cmax 40.8 μg/mL). In dog 28-day study, lesions in multiple organs were found at 100 mg/kg/day; NOAEL was 30 mg/kg/day (AUC 72.3 μg·mL⁻¹·h, Cmax 15.8 μg/mL). [1] Preclinical and clinical toxicology studies of GSK-1482160 have been conducted. In preclinical studies, the compound shows a favorable safety profile with no significant adverse effects at therapeutic doses. In healthy human subjects, pharmacokinetic and pharmacodynamic profiling has been performed. No significant organ toxicity or hematological abnormalities are reported at pharmacological doses. The compound shows no evidence of genotoxicity in standard in vitro assays. Cardiotoxicity risk appears low based on hERG channel inhibition studies. The safety profile supports further clinical development for inflammatory pain, neuroinflammation, and neurodegenerative diseases. The compound is for research use only and is not approved for human therapeutic use outside of clinical trials. |
| References | |
| Additional Infomation |
GSK1482160 (N-[(2-chloro-3-(trifluoromethyl)phenyl]methyl)-1-methyl-5-oxo-L-prolinamide) is an orally available negative allosteric modulator of the P2X7 receptor. It readily crosses the blood-brain barrier and is not a substrate for known active efflux transporters. The compound was progressed into clinical development for inflammatory and neuropathic pain indications. However, based on PK/PD modeling and simulation, it was determined that it was not possible to achieve the level of pharmacology (>90% inhibition of IL-1β release throughout the entire dosing interval) considered necessary to test the P2X7 mechanism adequately while maintaining a sufficient safety margin. This led to the decision to terminate development of GSK1482160 for chronic inflammatory pain indication. The therapeutic relevance of P2X7 receptor modulation remains to be established. [1]
GSK-1482160 is an orally available, brain-penetrant negative allosteric modulator of the P2X7 receptor. It is being evaluated for the treatment of inflammatory pain, neuroinflammation, and neurodegenerative diseases. The compound reduces ATP potency at the receptor and inhibits IL-1β release. It can be used as a radiolabeled probe for P2X7 receptor imaging. GSK-1482160 has been evaluated in healthy human subjects. It is not approved for general human use and is intended for research and clinical investigation purposes only. |
| Molecular Formula |
C14H14CLF3N2O2
|
|---|---|
| Molecular Weight |
334.721373081207
|
| Exact Mass |
334.07
|
| Elemental Analysis |
C, 50.24; H, 4.22; Cl, 10.59; F, 17.03; N, 8.37; O, 9.56
|
| CAS # |
1001389-72-5
|
| Related CAS # |
GSK-1482160 (isomer);1695551-19-9
|
| PubChem CID |
23649427
|
| Appearance |
White to off-white solid powder
|
| LogP |
2.924
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
22
|
| Complexity |
444
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
CN1[C@@H](CCC1=O)C(=O)NCC2=C(C(=CC=C2)C(F)(F)F)Cl
|
| InChi Key |
BJEMSIVBBUBXMZ-JTQLQIEISA-N
|
| InChi Code |
InChI=1S/C14H14ClF3N2O2/c1-20-10(5-6-11(20)21)13(22)19-7-8-3-2-4-9(12(8)15)14(16,17)18/h2-4,10H,5-7H2,1H3,(H,19,22)/t10-/m0/s1
|
| Chemical Name |
(2S)-N-[[2-chloro-3-(trifluoromethyl)phenyl]methyl]-1-methyl-5-oxopyrrolidine-2-carboxamide
|
| Synonyms |
GSK-1482160; GSK 1482160; GSK1482160; T1V3OH20HG; GSK1482160A;
|
| 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 : ~100 mg/mL (~298.76 mM)
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9876 mL | 14.9379 mL | 29.8757 mL | |
| 5 mM | 0.5975 mL | 2.9876 mL | 5.9751 mL | |
| 10 mM | 0.2988 mL | 1.4938 mL | 2.9876 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
