| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| Other Sizes |
| Targets |
GLP-1 receptor (GLP-1R). Exendin-4 (sequence: GTFTSDVSKQMEEEAVRLFIEWLKNGGPSSGAPPPS) is a 39-amino acid peptide that acts as a potent and selective agonist of the glucagon-like peptide-1 receptor (GLP-1R), a class B G protein-coupled receptor (GPCR). Upon binding to GLP-1R, exendin-4 activates Gs protein, leading to increased intracellular cAMP levels. This in turn activates PKA and Epac2, leading to glucose-stimulated insulin secretion (GSIS) from pancreatic beta-cells. Exendin-4 also suppresses glucagon secretion from pancreatic alpha-cells, delays gastric emptying, promotes satiety, and induces beta-cell proliferation and survival. Unlike native GLP-1, exendin-4 is resistant to dipeptidyl peptidase-4 (DPP-4) cleavage, providing a longer plasma half-life (2-4 hours in humans vs. 1-2 minutes for GLP-1). This makes exendin-4 a clinically useful GLP-1R agonist for the treatment of type 2 diabetes and obesity.
|
|---|---|
| ln Vitro |
Exendin-4 is a pure agonist of the GLP-1 receptor. Structurally generated from exendin-4, exendin-4 peptide derivatives may have a connection to dual GLP-1/glucagon receptor agonists. Their usage in medicine, for instance, includes reducing excessive food consumption and treating metabolic syndrome illnesses like diabetes and obesity. In order to lower the risk of hypoglycemia, these dual GLP-1/glucagon receptor agonists exhibit decreased action on the GIP receptor[1].
In vitro, exendin-4 (GTFTSDVSKQMEEEAVRLFIEWLKNGGPSSGAPPPS) is a potent GLP-1R agonist. In HEK293 cells stably expressing human GLP-1R, exendin-4 (0.001-100 nM) induces cAMP accumulation with an EC50 in the low pM to low nM range (typically 0.1-1 nM), similar to or slightly more potent than GLP-1(7-36) amide. In isolated rodent and human pancreatic islets, exendin-4 (0.1-100 nM) stimulates glucose-stimulated insulin secretion (GSIS) in a concentration-dependent manner, with maximal effect at 10-100 nM. It also protects beta-cells from apoptosis induced by cytokines (IL-1beta, TNF-alpha, IFN-gamma) or ER stress, and promotes beta-cell proliferation. In INS-1 or MIN6 beta-cell lines, exendin-4 (1-100 nM) increases PDX-1 expression, enhances insulin gene transcription, and activates PKA and PI3K/AKT pathways. In rodent and human neuronal cell lines, exendin-4 has neuroprotective effects (reducing oxidative stress, inflammation). In cell viability assays, exendin-4 is not cytotoxic at concentrations up to 100 nM. The compound is used as a positive control in GLP-1R signaling assays. Exendin-4 is the active pharmaceutical ingredient of the diabetes drug exenatide (Byetta, Bydureon). |
| ln Vivo |
In vivo, exendin-4 (GTFTSDVSKQMEEEAVRLFIEWLKNGGPSSGAPPPS) is the active ingredient of exenatide, an FDA-approved drug for type 2 diabetes and obesity. In animal models (e.g., diabetic db/db mice, STZ-induced diabetic rats, high-fat diet-fed mice), subcutaneous administration of exendin-4 (0.1-10 ug/kg, twice daily) reduces fasting and postprandial blood glucose levels, improves glucose tolerance, and lowers HbA1c. It also reduces body weight and food intake, and improves lipid profiles (triglycerides, cholesterol). In models of non-alcoholic fatty liver disease (NAFLD), exendin-4 reduces hepatic steatosis and inflammation. In models of neurodegenerative diseases (e.g., Parkinson's disease, Alzheimer's disease), exendin-4 (0.1-1 ug/kg/day, i.p. or i.c.v.) shows neuroprotective effects, reducing alpha-synuclein aggregation, amyloid-beta burden, and neuroinflammation, and improving cognitive function. In clinical studies in humans, exenatide (the synthetic version of exendin-4) improves glycemic control, reduces body weight, and has been shown to have cardiovascular benefits. It is approved for type 2 diabetes as an adjunct to diet and exercise. The research-grade peptide is used for preclinical studies. The TFA salt is used for solubility; in clinical formulations, the acetate salt is used.
|
| Enzyme Assay |
For non-cellular binding assays, surface plasmon resonance (SPR) can be used to measure the binding affinity of exendin-4 to the GLP-1 receptor extracellular domain. Immobilize recombinant human GLP-1R extracellular domain (ECD) on a CM5 sensor chip via amine coupling (EDC/NHS). Dissolve exendin-4 TFA in running buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.005% P20, 1 mM DTT) at concentrations of 0.1-100 nM. Flow over the immobilized ECD at 25degC at a flow rate of 30 uL/min. Record association (2-3 min) and dissociation (5-10 min) phases. Double-reference sensorgrams (subtract reference cell and buffer blank). Calculate the KD by fitting to a 1:1 Langmuir binding model. The KD for exendin-4 binding to GLP-1R is in the low nM range (e.g., 0.1-1 nM). For a competitive radioligand binding assay, incubate membranes from GLP-1R-expressing cells (e.g., HEK293-GLP-1R) with 0.05-0.1 nM 125I-GLP-1(7-36) amide and varying concentrations (0.01-1000 nM) of unlabeled exendin-4 in binding buffer (50 mM HEPES pH 7.4, 1 mM CaCl2, 5 mM MgCl2, 0.5% BSA, 0.1% bacitracin) for 60-90 minutes at 25degC. Separate bound and free by filtration through GF/B filters presoaked in 0.3% PEI. Count bound radioactivity. IC50 is determined, and Ki is calculated. The Ki for exendin-4 is typically 0.1-1 nM. For a functional cell-free assay, measure cAMP accumulation in membrane preparations: incubate GLP-1R-containing membranes with exendin-4 (0.1-100 nM) and GTPgammaS, then measure cAMP by ELISA. This is less common; cell-based assays are preferred.
|
| Cell Assay |
For cellular assays, use HEK293 cells stably expressing human GLP-1R. Seed cells in 96-well plates (2-4 × 10^4 cells/well) in DMEM with 10% FBS and culture overnight at 37degC, 5% CO2. On the assay day, replace medium with serum-free DMEM containing 0.5 mM IBMX (phosphodiesterase inhibitor) and pre-incubate for 20 minutes at 37degC. Add exendin-4 TFA at varying concentrations (0.0001-1000 nM) and incubate for 30 minutes at 37degC. Lyse cells and measure cAMP levels using an HTRF or chemiluminescence-based cAMP detection kit. The EC50 is calculated from a four-parameter logistic fit (typically 0.1-1 nM). For insulin secretion assays, use INS-1 rat insulinoma cells or isolated mouse islets. Seed cells in 24-well plates (2-4 × 10^5 cells/well) and culture in RPMI-1640 with 10% FBS. Starve cells in low-glucose (2.8 mM) KRB buffer for 1-2 hours, then incubate with exendin-4 (0.1-100 nM) in KRB buffer containing 2.8 or 16.7 mM glucose for 1 hour at 37degC. Collect supernatants and measure insulin by ELISA. Exendin-4 should increase insulin secretion only at the high glucose concentration. For proliferation assays, culture INS-1 cells for 48-72 hours in the presence of exendin-4 (1-100 nM). Measure cell number by MTT or CellTiter-Glo. For Western blot analysis of signaling, treat cells with exendin-4 (1-100 nM) for 5-60 minutes, lyse, and blot for p-AKT (Ser473), p-ERK (Thr202/Tyr204), p-CREB (Ser133). Control: vehicle (water or 0.1% DMSO). Positive control: GLP-1(7-36) amide (10 nM). The TFA salt is soluble in water or PBS. Prepare 0.1-1 mM stock and store at -80degC. Avoid repeated freeze-thaw cycles. All experiments should be performed in triplicate with at least three independent experiments.
|
| Animal Protocol |
For in vivo studies, use male C57BL/6J mice or db/db diabetic mice (8-12 weeks old). For acute glucose-lowering studies, fast mice for 6-16 hours. Dissolve exendin-4 TFA in sterile saline or PBS. Administer subcutaneously (s.c.) at doses of 0.1, 1, and 10 ug/kg (volume 5-10 mL/kg) 15-30 minutes before an intraperitoneal glucose tolerance test (IPGTT) (glucose 1-2 g/kg). Measure blood glucose at 0, 15, 30, 60, 90, 120 min. Exendin-4 should reduce glucose excursions. For chronic studies (2-4 weeks), administer exendin-4 (0.1-1 ug/kg, s.c., twice daily) to db/db mice or high-fat diet-fed mice. Monitor body weight, food intake, and blood glucose weekly. At termination, measure plasma insulin, HbA1c, triglycerides, and cholesterol. For neuroprotection studies (e.g., MPTP model of Parkinson's disease), administer exendin-4 (1-5 ug/kg, i.p., daily) for 1-2 weeks. Assess motor function (rotarod, pole test) and dopamine neuron counts (tyrosine hydroxylase staining). All animal procedures require IACUC approval.
|
| ADME/Pharmacokinetics |
Exendin-4 (exenatide) is a GLP-1 receptor agonist with a longer half-life than native GLP-1 due to its resistance to DPP-4 degradation. After subcutaneous administration in humans, the plasma half-life is 2-4 hours, enabling twice-daily (Byetta) or once-weekly (Bydureon) dosing. In rodents, the half-life is shorter (30-60 minutes). The compound is absorbed slowly from the subcutaneous injection site, with peak plasma concentrations (Cmax) reached at 1-2 hours (Tmax). It is cleared by the kidneys (glomerular filtration) and by proteolytic degradation. Exendin-4 is primarily excreted in urine. The TFA salt is used for research; the clinical formulation uses acetate salt. For PK studies in mice, administer exendin-4 (1-10 ug/kg, s.c.), collect blood at 0, 0.5, 1, 2, 4, 6, 8, 12 hours, and quantify by ELISA (specific for exendin-4) or LC-MS/MS. PK parameters (Cmax, Tmax, AUC, t1/2) are calculated. Exendin-4 has a large volume of distribution (Vd) due to tissue binding.
|
| Toxicity/Toxicokinetics |
Exenatide (synthetic exendin-4) has been extensively studied in preclinical and clinical toxicology. The most common adverse effects are gastrointestinal (nausea, vomiting, diarrhea), which are dose-dependent and mechanism-based. In preclinical studies in rodents and monkeys, exenatide at high doses (≥10 ug/kg/day for 2 weeks) may cause mild GI effects and transient weight loss. No genotoxicity, carcinogenicity (except thyroid C-cell tumors in rodents at high doses), or reproductive toxicity has been reported. The TFA salt is not used in clinical formulations; the acetate salt is used. For research, the TFA salt is acceptable for in vivo use but should be neutralized to pH 7.0-7.5. Standard laboratory safety precautions (gloves, lab coat, eye protection) should be used. The product is for research use only; clinical use requires a licensed formulation of exenatide.
|
| References |
[1]. US20150315260 A1
|
| Additional Infomation |
Exendin-4 is a 39-amino acid peptide isolated from the saliva of the Gila monster (Heloderma suspectum). It shares 53% homology with human GLP-1 but is resistant to DPP-4 degradation, making it a longer-acting GLP-1R agonist. Exendin-4 (exenatide) was the first GLP-1 receptor agonist approved by the FDA (2005) for the treatment of type 2 diabetes. It is also approved for chronic weight management (by reducing appetite) in combination with other agents. Exenatide is available as a twice-daily injection (Byetta) and a once-weekly extended-release formulation (Bydureon). The product is indicated as an adjunct to diet and exercise to improve glycemic control in adults with type 2 diabetes. It is not a first-line therapy but is used in patients who cannot achieve glycemic targets with metformin or other oral agents. Common side effects include nausea, vomiting, diarrhea, and injection site reactions. It is contraindicated in patients with a personal or family history of medullary thyroid carcinoma. This product is a research-grade peptide and is not for human use without a prescription. The TFA salt is for research only; clinical exenatide is the acetate salt.
|
| Molecular Weight |
3850.31
|
|---|---|
| Appearance |
Typically exists as solid at room temperature
|
| 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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
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
|
|---|---|
| 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 | 0.2597 mL | 1.2986 mL | 2.5972 mL | |
| 5 mM | 0.0519 mL | 0.2597 mL | 0.5194 mL | |
| 10 mM | 0.0260 mL | 0.1299 mL | 0.2597 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
