| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Adiponectin receptor (AdipoR). The compound is an agonist of AdipoR, which leads to the activation of multiple downstream signaling pathways.
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| ln Vitro |
AdipoR agonist 1 activates AMPK (AMP-activated protein kinase), SIRT1 (sirtuin 1), PPAR (peroxisome proliferator-activated receptors), and PGC-1alpha (peroxisome proliferator-activated receptor gamma coactivator 1alpha) through AdipoR signaling. The compound is a tool for studying the effects of AdipoR activation. The specific EC₅0 values for these activations are not provided in the available data. A general cell-free protocol for AdipoR binding is not commonly available due to the difficulty of expressing and purifying the full-length seven-transmembrane domain receptor. Alternative approaches include the use of radioligand binding assays on membrane preparations from cells overexpressing AdipoR. HEK293 cells stably expressing AdipoR1 or AdipoR2 are harvested and homogenized. Cell membranes are prepared by ultracentrifugation. The membrane pellets are resuspended in binding buffer (25 mM HEPES, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.5% BSA). 50 ug of membrane protein is incubated with 1 nM [¹2⁵I]-Adiponectin (or a fluorescently labeled AdipoR ligand) and varying concentrations of AdipoR agonist 1 (0.1 nM to 100 uM) in a total volume of 200 uL at room temperature for 60 minutes. Non-specific binding is determined in the presence of 10 uM unlabeled adiponectin or an excess of the unlabeled test compound. The reaction is terminated by rapid filtration through GF/B filters using a cell harvester. The filters are washed three times with ice-cold wash buffer, and the bound radioactivity is counted using a gamma counter. The IC₅0 is calculated, and the Kᵢ is derived using the Cheng-Prusoff equation.
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| Enzyme Assay |
A general cellular protocol for assessing AdipoR activation and AMPK phosphorylation: C2C12 mouse myoblasts are differentiated into myotubes in DMEM containing 2% horse serum for 5-7 days. The differentiated myotubes are serum-starved for 4 hours and then treated with various concentrations of AdipoR agonist 1 (0.1, 1, 10, 100 uM) for 15, 30, 60, and 120 minutes. The cells are then lysed in RIPA buffer supplemented with protease and phosphatase inhibitors. The lysates are centrifuged, and the supernatant is collected. Protein concentrations are quantified using the BCA assay. Equal amounts of protein (30 ug) are separated by SDS-PAGE and transferred to PVDF membranes. The membranes are probed with antibodies against phospho-AMPKalpha (Thr172), total AMPKalpha, phospho-ACC (Ser79), total ACC, and GAPDH (loading control). The blots are developed using an ECL chemiluminescence kit. The ratio of phospho-AMPK to total AMPK and phospho-ACC to total ACC is quantified by densitometry. For SIRT1 activation, HEK293T cells are transfected with a pGL4.32[luc2P/NF-kappaB-RE/Hygro] reporter plasmid and a SIRT1 expression plasmid. After 24 hours, the cells are treated with AdipoR agonist 1 (1-100 uM) for 24 hours, and luciferase activity is measured.
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| Animal Protocol |
A general animal protocol for evaluating AdipoR activation and energy metabolism: Male C57BL/6J mice (8 weeks old) are fed a high-fat diet (HFD, 60% kcal from fat) for 12 weeks to induce obesity and insulin resistance. The mice are then randomized into treatment groups (n=10/group). AdipoR agonist 1 is formulated in a suitable vehicle (e.g., 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline) and administered via intraperitoneal (IP) injection at doses of 5, 10, and 20 mg/kg once daily for 28 days. A vehicle control group and a positive control group (e.g., metformin at 200 mg/kg) are included. Body weight and food intake are measured weekly. An oral glucose tolerance test (OGTT) is performed after 14 days of treatment. Mice are fasted for 6 hours, and blood glucose is measured at 0, 15, 30, 60, 90, and 120 minutes after oral administration of 2 g/kg glucose. Insulin levels are measured at 0 and 30 minutes by ELISA. At the end of the study, mice are euthanized, and blood is collected for serum chemistry (triglycerides, total cholesterol, NEFA, adiponectin). Tissues (liver, epididymal white adipose tissue, skeletal muscle) are harvested, weighed, and processed for histology (H&E and Oil Red O staining). Protein and RNA are extracted from these tissues for Western blot analysis (AMPK, ACC, SIRT1, PGC-1alpha, PPARgamma) and qRT-PCR (PPARgamma target genes, PGC-1alpha, CPT-1).
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| ADME/Pharmacokinetics |
General pharmacokinetic protocol for AdipoR agonist 1: Male Sprague-Dawley rats are administered AdipoR agonist 1 via oral gavage (PO, 10 mg/kg) and intravenous (IV, 2 mg/kg) injection. The compound is formulated in a vehicle such as 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline. Blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours post-dose. Plasma concentrations of AdipoR agonist 1 are quantified by LC-MS/MS. PK parameters (Cmax, Tmax, AUC, t½, clearance, Vd, and oral bioavailability) are calculated using non-compartmental analysis. The compound has a molecular weight of 437.57 and a formula of C2₆H3₅N3O3. It is soluble in DMSO at 5 mg/mL (11.43 mM).
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| Toxicity/Toxicokinetics |
General toxicity protocol for AdipoR agonist 1: A 14-day repeated-dose toxicity study is performed in ICR mice. AdipoR agonist 1 is administered via intraperitoneal (IP) injection at doses of 10, 30, and 100 mg/kg/day for 14 consecutive days. Clinical signs, body weight, and food consumption are monitored daily. At the end of the study, blood samples are collected for hematology (complete blood count with differential) and serum chemistry (ALT, AST, ALP, BUN, creatinine, total protein, albumin, glucose, triglycerides, cholesterol). Gross necropsy is performed, and the weights of major organs (liver, kidney, spleen, heart, lung, brain, adipose tissue, muscle) are recorded. Histopathological examination of these organs is conducted. Special attention is given to the heart and liver, as AMPK and SIRT1 are important regulators of cardiac and hepatic function.
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| References | |
| Additional Infomation |
AdipoR agonist 1 (Compound 112254) is an agonist of the adiponectin receptor. The molecular formula is C2₆H3₅N3O3, and the molecular weight is 437.57. The IUPAC name is 4-tert-butyl-N-(3-{4-[(4-methoxyphenyl)methyl]piperazin-1-yl}-3-oxopropyl)benzamide. The compound is soluble in DMSO at 5 mg/mL. It is stored at -20degC. AdipoR agonist 1 is utilized in preventive doping research. It is a research tool for studying the adiponectin signaling pathway, which is involved in the regulation of glucose and lipid metabolism, insulin sensitivity, and energy homeostasis.
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| Molecular Formula |
C26H35N3O3
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|---|---|
| Molecular Weight |
437.57
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| Exact Mass |
437.267
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| CAS # |
949745-75-9
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| PubChem CID |
17528991
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| Appearance |
White to off-white solid powder
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| Density |
1.1±0.1 g/cm3
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| Boiling Point |
624.4±55.0 °C at 760 mmHg
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| Flash Point |
331.4±31.5 °C
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| Vapour Pressure |
0.0±1.8 mmHg at 25°C
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| Index of Refraction |
1.564
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| LogP |
3.71
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
32
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| Complexity |
594
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC(C)(C)C1=CC=C(C=C1)C(=O)NCCC(=O)N2CCN(CC2)CC3=CC=C(C=C3)OC
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| InChi Key |
XBEMZEZVTNPLQI-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H35N3O3/c1-26(2,3)22-9-7-21(8-10-22)25(31)27-14-13-24(30)29-17-15-28(16-18-29)19-20-5-11-23(32-4)12-6-20/h5-12H,13-19H2,1-4H3,(H,27,31)
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| Chemical Name |
4-tert-butyl-N-[3-[4-[(4-methoxyphenyl)methyl]piperazin-1-yl]-3-oxopropyl]benzamide
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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)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| 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 | 2.2853 mL | 11.4267 mL | 22.8535 mL | |
| 5 mM | 0.4571 mL | 2.2853 mL | 4.5707 mL | |
| 10 mM | 0.2285 mL | 1.1427 mL | 2.2853 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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