1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEMRK-016Cat. No.: HY-100370CAS No.: 342652-67-9分式: CHNO分量: 368.39作靶點(diǎn): GABA Receptor作通路: Membrane Transporter/Ion Channel; Neuronal Signaling儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 50

2、 mg/mL (135.73 mM; Need ultrasonic)Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 2.7145 mL 13.5726 mL 27.1451 mL5 mM 0.5429 mL 2.7145 mL 5.4290 mL10 mM 0.2715 mL 1.3573 mL 2.7145 mL請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲(chǔ)備液，并請(qǐng)注意儲(chǔ)備液的保存式和期限。體內(nèi)實(shí)驗(yàn) 請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)?shù)娜芙獍福渲魄罢?qǐng)先配制澄清的儲(chǔ)備液，再依次添加助溶劑(為保證實(shí)驗(yàn)結(jié)果的可靠性，體內(nèi)實(shí)驗(yàn)的作液，

3、建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄清的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；以下溶劑前的百分 指該溶劑在您配制終溶液中的體積占)：1. 請(qǐng)依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.75 mg/mL (7.46 mM); Clear solution2. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.75 mg/mL (7.46 mM); Clear solution3. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubi

4、lity: 2.75 mg/mL (7.46 mM); Clear solution1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEBIOLOGICAL ACTIVITY物活性 MRK-016種選擇性的，可服的，可透過(guò)腦屏障的 GABAA 5 receptor 拮抗劑，對(duì) GABAA 5 的EC50 值為 3 nM，對(duì) GABAA 132，GABAA 232，GABAA 332 和 GABAA 532 的Ki 值分別為 0.83，0.85，0.77 和 1.4 nM。IC50 & Target EC50: 3 nM (GABAA 5) 1Ki:

5、 0.83nM (HumanGABAA132), 0.85nM (HumanGABAA232), 0.77nM (HumanGABAA332),1.4nM (HumanGABAA532) 1體外研究 MRK-016 is a selective, orally bioavailable inverse agonist of GABAA 5 receptor, with an EC50 of 3 nM forGABAA 5, and Kis of 0.83, 0.85, 0.77and 1.4nM for humanGABAA132, GABAA232, GABAA332, and GABAA5

6、32, respectively 1 2. MRK-016 is a full inverse agonist at the 5-subtype, showsvery weak affinity at the GABAA 432-subtype (Ki 395 173 nM) and is essentially inactive at the GABAA632 receptor (Ki 4000 nM) 1. MRK-016 shows a weak effect on GABAA432 with a Ki of 400 nM.MRK-016 (100 nM) alao increases

7、long-term potentiation in mouse hippocampal slices 2.體內(nèi)研究 MRK-016 does not enhance pentylenetetrazole-induced convulsions at 10 mg/kg via ip, or cause seizures at30 mg/kg, via po for 20 days in mice. MRK-016 shows no obvious anxiogenic-like effects in rats at doses thatoccupy 95% of benzodiazepine (

8、BZ) binding sites. MRK-016 (0.3, 1, and 3 mg/kg, p.o.) dose-dependentlyimproves performance of rats hippocampal-dependent memory task 1. MRK-016 (0.3-30 mg/kg, p.o.)causes good receptor occupancy in rats. MRK-016 (0.3, 1, or 3 mg/kg p.o.) shows cognition-enhancingactivity in the delayed matching-to-

9、position version of the Morris water maze. MRK-016 (1, 3, or 10 mg/kgi.p.) does not produce kindling in mice 2. MRK-016 (3 mg/kg, i.p.) protects against LPS-inducedlearning/memory decrements in mice 3.PROTOCOLKinase Assay 1 L(tk-) cells expressing human recombinant GABAA receptors containing 3- and

10、2-subunits in combinatioonwith various -subunits are harvested and binding performed. The displacement of 3HRo 15-1788 bindingby the test compounds (MRK-016, etc.) is measured in GABAA receptors containing eigher an 1-, 2-, 3-,and 5-subunit and from the IC50 and the Ki is calculated assuming respect

11、ive Kd values of 3HRo 15-1788binding of 0.92, 1.05, 0.58 and 0.45 nM at the 1-, 2-, 3-, and 5-subtypes. Nonspecific binding is definedby the inclusion of 10 M flunitrazepam for the 1-, 2-, 3-, and 5-subtypes. The percentage inhibition of3HRo 15-1788, the IC50 and the Ki values are calculated using A

12、ctivityBase 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 1Administration 1 The proconvulsant potential MRK-016 is determined in this assay. The convulsant is pentylenetetrazolewhich is dosed at 15 mg/mL with an infusion rate of 0.2 mL/m

13、in. This rate is chosen to ensure that drug-naivemince reach the terminal convulsion sign within 1 min. MRK-016 is dosed intraperitoneally (ip) at aconcentration of 1, 3, and 10 mg/kg in 70% PEG 300 1.2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemEMCE has not independently confirmed the accuracy o

14、f these methods. They are for reference only.REFERENCES1. Chambers MS, et al. An orally bioavailable, functionally selective inverse agonist at the benzodiazepine site of GABAA alpha5 receptorswith cognition enhancing properties. J Med Chem. 2004 Nov 18;47(24):5829-32.2. Atack JR, et al. In vitro an

15、d in vivo properties of 3-tert-butyl-7-(5-methylisoxazol-3-yl)-2-(1-methyl-1H-1,2,4-triazol-5-ylmethoxy)-pyrazolo1,5-d-1,2,4triazine (MRK-016), a GABAA receptor alpha5 subtype-selective inverse agonist. J Pharmacol Exp Ther. 2009Nov;331(2):470-84.3. Eimerbrink MJ, et al. Administration of the inverse benzodiazepine agonist MRK-016 rescues acquisition and memory consolidationfollowing peripheral administration of bacterial endotoxin. Behav