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| 25mg |
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Purity: ≥98%
GRA Ex-25 is a novel and potent inhibitor of glucagon receptor with IC50 of 56 and 55 nM for rat and human glucagon receptors, respectively. In our assay conditions, GRA Ex-25 binds a human glucagon receptor (h-GlucRbind) with a Ki of 63 nM and a moderate glucagon-induced adenylate cyclase inhibition (h-GlucRcyclase) with a Ki of 254 nM. GRA Ex-25 exhibits a comparable affinity (IC50=56 and 55 nM, respectively) for the rat and human glucagon receptors. When glucagon is administered exogenously to rats, GRA Ex-25 (3 mg/kg, i.v.) dramatically lowers blood glucose levels. Thanks primarily to its direct inhibition of glucagon-stimulated hepatic glucose output, GRA Ex-25 can block the rise in blood glucose levels caused by exogenous administered glucagon.
| Targets |
rat glucagon receptor ( IC50 = 56 nM ); human glucagon receptor ( IC50 = 55 nM )
Glucagon Receptor [1] Glucagon Receptor (initial hit compound targeting this receptor had an IC50 of 7 μM; optimized derivatives including GRA Ex-25 achieved nanomolar affinity for the receptor) [2] |
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| ln Vitro |
GRA Ex-25 exhibits a 63 nM Ki for the human glucagon receptor (h-GlucRbind) and a 254 nM Ki for the moderate glucagon-induced adenylate cyclase inhibition (h-GlucRcyclase) in our assay[1]. The affinity of GRA Ex-25 for the rat and human glucagon receptors is comparable (IC50=56 and 55 nM, respectively)[2].
1. GRA Ex-25 (a β-alanine derivative) was optimized from an initial weak human glucagon receptor antagonist (IC50 = 7 μM) via structure-activity relationship studies to enhance binding affinity to the glucagon receptor, resulting in nanomolar-level receptor binding affinity; optimization was also performed to improve metabolic stability of the molecule against fast metabolic turnover [2] |
| ln Vivo |
GRA Ex-25 (3 mg/kg, i.v.) dramatically lowers blood glucose brought on by exogenous glucagon administration in rat model. Because GRA Ex-25 directly inhibits glucagon-stimulated hepatic glucose output, it can prevent the rise in blood glucose levels brought on by exogenous glucagon administration[2].
1. GRA Ex-25 (a β-alanine derivative) exhibited 17% oral bioavailability in rats with a plasma half-life of 90 min; the final optimized glucagon antagonist (including GRA Ex-25) was orally available in rats and dogs, and demonstrated efficacy in lowering blood glucose levels in a diabetic animal model [2] |
| Enzyme Assay |
1. Receptor binding assay for glucagon receptor: A library of compounds (initially designed to mimic GLP-1 hormone binding to its receptor) was screened to identify potential glucagon receptor antagonists. The binding affinity of the initial hit compound and optimized derivatives (including GRA Ex-25) to the human glucagon receptor was measured using a receptor binding assay; the assay evaluated the ability of compounds to compete with glucagon for binding to the receptor, and the IC50 value of the initial hit was determined to be 7 μM. Subsequent optimization efforts focused on improving binding affinity, leading to nanomolar affinity for GRA Ex-25 [2]
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| Animal Protocol |
Male Sprague Dawley rats weighing 200 grams who have not fasted are kept anesthetized throughout the test by intraperitoneal injection of a 1:1 combination of Hypnorm (fentanyl, 0.05 mg/mL and fluanizone, 2.5 mg/mL) and Dormicum (midazolam, 1.25 mg/mL). In order to administer compounds, an catheter is placed in the jugular vein. Test compounds (0, 1, 3, 10, and 30 mg/kg) and glucagon (3 μg/kg) are given in intervals of 5 minutes, approximately 60 minutes after the anesthesia is started. Samples for determination of blood glucose concentrations are taken from the tail tip 25 and 5 min prior to administration of the compound to represent average basal values and again 10 min after administration of glucagon (time for peak response of glucagon). The values obtained 10 minutes after alucagon administration are subtracted from the average of the two basal values to express the results as delta values.
1. Oral administration protocol for GRA Ex-25 in rats: GRA Ex-25 (formulated in a suitable oral dosage form, specific formulation not detailed) was administered orally to rats; plasma samples were collected at predetermined time points to measure plasma concentration of the drug, and pharmacokinetic parameters (oral bioavailability 17%, plasma half-life 90 min) were calculated. For the diabetic animal model study: GRA Ex-25 (final optimized formulation, specific dosage and frequency not detailed) was administered orally to diabetic animals (type not specified), and blood glucose levels were monitored at regular intervals to evaluate the hypoglycemic efficacy of the drug. The optimized glucagon antagonist (including GRA Ex-25) was also administered orally to dogs (dosage and frequency not specified) to assess oral availability [2] |
| ADME/Pharmacokinetics |
1. The oral bioavailability of GRA Ex-25 (a β-alanine derivative) in rats was 17%.
The plasma half-life of GRA Ex-25 in rats was 90 minutes. The major metabolites of GRA Ex-25 have been identified, and the pharmacokinetic properties of a series of β-alanine derivatives have been further optimized using this information [2]. |
| References | |
| Additional Infomation |
1. GRA Ex-25 is a β-alanine derivative that was developed as a glucagon receptor antagonist. The initial lead compound was discovered by screening a library of compounds that mimic the binding of GLP-1 hormone to its receptor, and its glucagon receptor antagonism was weak (IC50 = 7 μM). The optimization of the initial lead compound focused on improving its binding affinity and metabolic stability to the glucagon receptor, and finally GRA Ex-25 with nanomolar receptor affinity and better pharmacokinetic properties was obtained. The final optimized glucagon antagonists (including GRA Ex-25) have oral bioavailability in rats and dogs and can effectively reduce blood glucose in diabetic animal models, indicating their potential for treating type 2 diabetes [2].
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| Molecular Formula |
C29H36F3N3O5
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|---|---|
| Molecular Weight |
563.60844
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| Exact Mass |
563.261
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| Elemental Analysis |
C, 61.80; H, 6.44; F, 10.11; N, 7.46; O, 14.19
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| CAS # |
307983-31-9
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| PubChem CID |
16100296
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| Appearance |
Solid powder
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| LogP |
6.892
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
40
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| Complexity |
839
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(O)CCNC(C1=CC=C(CN([C@H]2CC[C@H](C(C)(C)C)CC2)C(NC3=CC=C(OC(F)(F)F)C=C3)=O)C=C1)=O
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| InChi Key |
BZXMLCVDKDXRQY-LOSWNTGBSA-N
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| InChi Code |
InChI=1S/C29H36F3N3O5/c1-28(2,3)21-8-12-23(13-9-21)35(27(39)34-22-10-14-24(15-11-22)40-29(30,31)32)18-19-4-6-20(7-5-19)26(38)33-17-16-25(36)37/h4-7,10-11,14-15,21,23H,8-9,12-13,16-18H2,1-3H3,(H,33,38)(H,34,39)(H,36,37)/t21?,23-
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| Chemical Name |
3-(4-((1-(4-(tert-butyl)cyclohexyl)-3-(4-(trifluoromethoxy)phenyl)ureido)methyl)benzamido)propanoic acid
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| Synonyms |
GRA Ex25; GRA Ex-25; GRAEx 25; GRA Ex 25; GRAEx-25; GRAEx25
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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) |
DMSO: ≥ 32 mg/mL (~56.8 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.44 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: ≥ 2.08 mg/mL (3.69 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 20.8 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: ≥ 2.08 mg/mL (3.69 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 | 1.7743 mL | 8.8714 mL | 17.7428 mL | |
| 5 mM | 0.3549 mL | 1.7743 mL | 3.5486 mL | |
| 10 mM | 0.1774 mL | 0.8871 mL | 1.7743 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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