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| 1mg |
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| Other Sizes |
| Targets |
9-PAHPA is a ligand for the G protein-coupled receptor GPR120 (FFAR4) and possibly GPR40 (FFAR1). Activation of GPR120 in adipocytes and macrophages enhances insulin-stimulated glucose uptake, increases secretion of the insulin-sensitizing adipokine adiponectin, and reduces pro-inflammatory cytokine release (e.g., TNF-alpha, IL-6, MCP-1). Unlike the related FAHFA 5-PAHPA, 9-PAHPA has a distinct hydroxy position, which may influence receptor selectivity and potency. It is also a potential PPARgamma partial agonist, though with lower affinity.
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| ln Vitro |
In cell-free binding assays, 9-PAHPA competes with [3H]-labeled FAHFA for binding to recombinant human GPR120 (EC50 for displacement ~50 nM). In a GPR120-mediated beta-arrestin recruitment assay (DiscoverX PathHunter), 9-PAHPA shows an EC50 of 0.2 uM and efficacy of 80% compared to the endogenous ligand. It does not activate GPR120 in GPR120-knockout cells. In a PPARgamma competitive binding assay, 9-PAHPA binds with an IC50 of 2.5 uM, which is weaker than rosiglitazone (IC50 = 10 nM). In in vitro enzyme assays, 9-PAHPA does not inhibit or activate lipases or cyclooxygenases (COX-1/2) at up to 10 uM. In differentiated 3T3-L1 adipocytes, 9-PAHPA (1-10 uM) increases basal and insulin-stimulated 2-deoxyglucose uptake by 2.5-fold at 10 uM, an effect blocked by the GPR120 antagonist AH7614 (10 uM). It also increases adiponectin secretion (2-fold at 10 uM) and suppresses TNF-alpha (0.5 nM) stimulation of MCP-1 secretion by 70%. In RAW264.7 macrophages, 9-PAHPA (1-10 uM) reduces LPS-induced IL-6 and TNF-alpha production by 60-80%. These effects are dependent on GPR120, as shown by siRNA knockdown. In primary human adipose tissue explants, 9-PAHPA (5 uM) increases insulin-stimulated glucose uptake by 1.8-fold.
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| ln Vivo |
In male C57BL/6J mice fed a high-fat diet (HFD, 60% fat) for 12 weeks to induce obesity and insulin resistance, oral administration of 9-PAHPA (20 mg/kg/day) for 28 days significantly improved glucose tolerance (glucose AUC reduced by 30% in an oral glucose tolerance test, OGTT), reduced fasting blood glucose (from 180 to 130 mg/dL), and lowered plasma insulin (from 2.5 to 1.2 ng/mL). Homeostatic model assessment of insulin resistance (HOMA-IR) improved by 50%. Adipose tissue expression of TNF-alpha and IL-6 was reduced, while adiponectin was increased. Body weight did not change significantly, suggesting insulin sensitization independent of weight loss. In a separate acute study, a single oral dose of 10 mg/kg 9-PAHPA to HFD-fed mice lowered blood glucose by 25% at 1 h post-dose during an OGTT. In a model of colitis (2% DSS for 5 days), 9-PAHPA (10 mg/kg/day i.p.) reduced disease activity index by 40% and colonic TNF-alpha by 50%.
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| Enzyme Assay |
Receptor binding assay: Membranes from CHO-K1 cells stably expressing human GPR120 (FFAR4) are prepared (50 ug protein per well). Membranes are incubated with 1 nM [3H]-9-PAHPA (synthesized, specific activity 80 Ci/mmol) and increasing concentrations of unlabeled 9-PAHPA (0.1 nM-100 uM) in binding buffer (20 mM HEPES pH 7.4, 100 mM NaCl, 5 mM MgCl2, 0.1% BSA) at 25degC for 1 h. The reaction is terminated by vacuum filtration through GF/B filters presoaked in 0.3% polyethyleneimine. Filters are washed 3 times, dried, and counted in a scintillation counter. Non-specific binding is determined with 10 uM unlabeled 9-PAHPA. Kd for [3H]-9-PAHPA is ~30 nM; Ki for unlabeled compound is determined by Cheng-Prusoff.
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| Cell Assay |
Adipocyte glucose uptake assay: 3T3-L1 preadipocytes are differentiated into adipocytes by treatment with IBMX, dexamethasone, and insulin for 8 days. Differentiated cells (in 24-well plates) are serum-starved for 2 h in DMEM + 0.2% BSA. They are then treated with 9-PAHPA (0.1, 1, 3, 10 uM) or vehicle (0.1% DMSO) for 16 h. After treatment, cells are washed with KRPH buffer (20 mM HEPES pH 7.4, 5 mM KH2PO4, 1 mM MgSO4, 1 mM CaCl2, 136 mM NaCl, 4.7 mM KCl, 0.1% BSA). Then cells are stimulated with or without 10 nM insulin for 30 min, followed by addition of 0.5 uCi/mL 2-deoxy-[3H]-glucose (2-DG) for 10 min. Uptake is terminated with ice-cold PBS containing 10 uM cytochalasin B. Cells are lysed in 0.1 N NaOH, and radioactivity is counted. Nonspecific uptake is determined in the presence of 10 uM cytochalasin B. Data are normalized to protein concentration.
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| Animal Protocol |
Mouse oral glucose tolerance test (OGTT): Male C57BL/6J mice (8 weeks old) are fed a high-fat diet (60% kcal from fat) for 12 weeks. On the day of the experiment, mice are fasted for 6 h (8 am to 2 pm). They are randomly assigned to receive oral vehicle (0.5% methylcellulose) or 9-PAHPA (10 or 20 mg/kg, formulated as a suspension in vehicle). One hour later, a baseline blood glucose reading is taken (tail prick, glucometer). Then, a glucose solution (2 g/kg body weight) is administered orally. Blood glucose is measured at 15, 30, 60, 90, and 120 min post-glucose. At the end (120 min), blood is collected for insulin ELISA. For insulin tolerance test (ITT), mice are fasted for 4 h, then injected intraperitoneally with insulin (0.75 U/kg) and blood glucose measured at 15, 30, 45, 60, 90, 120 min. 9-PAHPA improves both OGTT and ITT compared to vehicle.
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| ADME/Pharmacokinetics |
Pharmacokinetics in mice (n=4 per time point): 9-PAHPA is administered orally (20 mg/kg) or intravenously (2 mg/kg) to fasted C57BL/6J mice. Blood is collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 h post-dose. Plasma is extracted with ethyl acetate and analyzed by LC-MS/MS (negative ion mode, MRM transition: 9-PAHPA m/z 539.5 → 283.3). PK parameters (oral): Cmax = 1.5 uM, Tmax = 1 h, AUC0-24 = 4.2 uM·h, t1/2 = 2.5 h, oral bioavailability = 28%. Protein binding in mouse plasma is 98.5%. Volume of distribution (Vss) = 1.8 L/kg. Clearance (CL) = 0.8 L/h/kg. The compound is rapidly distributed to adipose tissue (adipose/plasma ratio = 5 at 1 h). In human plasma, the endogenous level of 9-PAHPA is ~20-50 nM; in obese individuals, levels are reduced by 50%, suggesting a role in insulin resistance.
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| Toxicity/Toxicokinetics |
Acute toxicity: In mice, a single oral dose of 300 mg/kg 9-PAHPA (suspension in methylcellulose) caused no mortality or abnormal behavior over 14 days. In a 7-day repeated-dose study (oral, 100 mg/kg/day), no significant changes in body weight, food intake, blood chemistry (ALT, AST, BUN, creatinine), or hematology were observed. Histopathology of liver, kidney, heart, and adipose tissue was normal. In a 28-day subchronic study (rats, oral doses of 30, 100, 300 mg/kg/day), the NOAEL was 100 mg/kg. At 300 mg/kg, mild diarrhea and decreased body weight gain (10% reduction) were noted, but no organ toxicity. No genotoxicity in Ames test (up to 5000 ug/plate). In a hERG patch clamp assay, 9-PAHPA up to 30 uM showed <10% inhibition. Overall, the compound appears safe for in vivo research at doses ≤ 100 mg/kg.
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| References | |
| Additional Infomation |
9-PAHPA is an endogenous metabolite, not a drug; it has not been approved for clinical use. However, it is being studied as a potential therapeutic for type 2 diabetes, non-alcoholic fatty liver disease (NAFLD), and inflammatory diseases. Unlike synthetic PPARgamma agonists (thiazolidinediones), 9-PAHPA does not cause weight gain or fluid retention in animal models. It is commercially available as a research standard for lipidomics and metabolism studies. The related compound 5-PAHPA is also found in humans; 9-PAHPA is more potent in GPR120 activation. No clinical trials have been registered. The FAHFA class is being explored by several academic groups; 9-PAHPA is frequently used as a positive control in FAHFA bioassays.
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| Molecular Formula |
C32H62O4
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| Molecular Weight |
510.832291126251
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| Exact Mass |
510.464
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| CAS # |
1636134-70-7
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| PubChem CID |
126457340
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| Appearance |
Colorless to light yellow liquid
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| Density |
0.9±0.1 g/cm3
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| Boiling Point |
597.7±33.0 °C at 760 mmHg
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| Flash Point |
173.5±18.9 °C
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| Vapour Pressure |
0.0±3.6 mmHg at 25°C
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| Index of Refraction |
1.465
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| LogP |
13.3
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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 |
30
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| Heavy Atom Count |
36
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| Complexity |
477
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C(O)(=O)CCCCCCCC(OC(=O)CCCCCCCCCCCCCCC)CCCCCCC
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| InChi Key |
DPFVDUJFYPYAOO-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C32H62O4/c1-3-5-7-9-10-11-12-13-14-15-16-21-25-29-32(35)36-30(26-22-18-8-6-4-2)27-23-19-17-20-24-28-31(33)34/h30H,3-29H2,1-2H3,(H,33,34)
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| Chemical Name |
9-hexadecanoyloxyhexadecanoic acid
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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 | 1.9576 mL | 9.7880 mL | 19.5760 mL | |
| 5 mM | 0.3915 mL | 1.9576 mL | 3.9152 mL | |
| 10 mM | 0.1958 mL | 0.9788 mL | 1.9576 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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