MK-3903

AMPK (EC50 = 8 nM)
MK-3903 (compound 42) is a potent and selective AMP-activated protein kinase (AMPK) activator with an EC50 of 8 nM. MK-3903 activates 10 of the 12 phosphorylated AMPK (pAMPK) complexes with EC50 values in the range of 8 to 40 nM and maximal activation >50%. MK-3903 does not activate pAMPK6 and only partially activates pAMPK5 (up to 36% maximum). In LLC-PK1 cells, MK-3903 exhibits low permeability (Papp=6 10-6 cm/s), and it is a substrate for the organic anion transporter proteins OATP1B1 and OATP1B3 that are found in human liver uptake transporters. According to the findings, MK-3903 moderately binds to the prostanoid DP2 (CRTH2) receptor (binding IC50=1.8 μM), but not when 10% human serum is present (binding IC50>86 μM)[1].

MK-3903 (compound 42) 是一种有效的选择性 AMP 激活蛋白激酶 (AMPK) 激活剂，EC50 为 8 nM。 MK-3903 可激活 12 种磷酸化 AMPK (pAMPK) 复合物中的 10 种，EC50 值在 8 至 40 nM 范围内，最大激活 >50%。 MK-3903 不会激活 pAMPK6，仅部分激活 pAMPK5（最多 36%）。在 LLC-PK1 细胞中，MK-3903 表现出低渗透性 (Papp=6 10-6 cm/s)，并且它是人肝脏摄取转运蛋白中发现的有机阴离子转运蛋白 OATP1B1 和 OATP1B3 的底物。根据研究结果，MK-3903 适度结合前列腺素 DP2 (CRTH2) 受体（结合 IC50=1.8 μM），但当存在 10% 人血清时则不会结合（结合 IC50>86 μM）[1]。

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MK-3903 在双向LLC-PK1细胞模型中表现出低渗透性 (Papp = 6 x 10⁻⁶ cm/s)。
它是人肝摄取转运蛋白OATP1B1和OATP1B3的底物。
它未表现出对CYP3A4活性的时间依赖性抑制。
在体外人孕烷X受体 (hPXR) 实验中，其EC₅₀ > 30 µM，表明它不是强效的PXR激动剂。[1]

MK-3903（化合物 42）具有中等的全身血浆清除率（5.0 至 13 mL/min/kg），稳态分布容积为 0.6 至 1.1 L/kg，在 C57BL/6 中的终末半衰期约为 2 小时小鼠、斯普拉格大鼠、道利大鼠和比格犬。当给高果糖喂养的 db/+ 小鼠急性口服给药时，MK-3903（3、10 和 30 mg/kg）会导致肝脏脂肪酸合成 (FAS) 显着减少[1]。

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在饮食诱导肥胖 (DIO) 小鼠中，口服MK-3903 (30 mg/kg, 每日两次，BID) 治疗15天后，末次给药后2小时，作为AMPK激活标志物的磷酸化ACC与总ACC比值 (pACC/ACC) 在肝脏中显著增加 (3.1倍)，在骨骼肌中显著增加 (1.6倍)。[1]
在高果糖喂养的db/+小鼠中，急性口服MK-3903 (3、10和30 mg/kg) 在所有剂量下均显著抑制肝脏脂肪酸合成 (FAS)，抑制效果在给药后2小时和8小时测定。在2小时，3、10和30 mg/kg剂量的抑制率分别为60%、61%和66%。在8小时，相应剂量的抑制率分别为29%、37%和49%。[1]
在DIO小鼠中，慢性口服MK-3903 (3-30 mg/kg BID 和 30 mg/kg QD) 治疗12天，导致口服葡萄糖耐量试验 (OGTT) 期间胰岛素抵抗指数 (IRI) 呈剂量依赖性降低，这主要是由于胰岛素分泌受到抑制。在10 mg/kg BID和30 mg/kg QD剂量下观察到对IRI的显著影响，且无显著体重变化。在30 mg/kg BID剂量下观察到轻微的体重减轻 (3.1%)。[1]
腹腔注射早期类似物化合物36 (30 mg/kg) 导致小鼠肝脏新生脂肪生成 (DNL) 抑制约40%。[1]

进行酶促反应。简而言之，为了产生 pAMPK，将目标 AMPK 复合物在 AMPK 反应缓冲液中适当稀释，并在室温下孵育 30 分钟。将 DMSO 中适当稀释的 MK-3903（化合物 42）（总共 1.2 L）添加到含有 pAMPK（每孔 15 L）的反应缓冲液中后，将板短暂涡旋，然后在室温下孵育 30分钟。室温孵育60分钟后，将板密封，并添加淬灭缓冲液以终止反应。产品与活化剂浓度图用于计算 EC50 和活化参数[1]。

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通过测量重组人AMPK复合物对合成肽底物 (SAMS) 的磷酸化来评估AMPK活性。特异性复合物pAMPK7 (α2β1γ1) 通过催化α亚基的磷酸化进行预激活。变构激活剂AMP作为最大激活的阳性对照。将测试化合物与酶和底物一起孵育，并测量活性。使用10点滴定法生成剂量反应曲线，以确定相对于AMP的EC₅₀值和最大激活百分比。[1]
使用类似的实验条件将激活谱分析扩展到所有12种重组人pAMPK复合物，以确定亚型选择性。[1]

In this study, DIO mice that are 17 weeks old are used. Mice are trained to receive vehicle at a dose of 5 mL/kg BID for 5 days (vehicle: 5% Tween 80, 0.25% methylcellulose, 0.02% SDS). At that point, mice are bled, their blood sugar and insulin levels are measured, and the mice are then divided into treatment groups according to body weight, blood sugar, and insulin levels. For a 12-day BID period, MK-3903 (compound 42) is administered orally to each group of animals at doses of 3 mg/kg, 10 mg/kg, 30 mg/kg, or just the vehicle. A second group of mice that received MK-3903 with vehicle at 30 mg/kg for 12 days is also included. Every day, both food intake and body weight are recorded[1].

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For chronic target engagement studies in diet-induced obese (DIO) mice, MK-3903 was administered orally at 30 mg/kg twice daily (BID) for 15 days. Animals were fasted for 4 hours prior to tissue collection. Liver and skeletal muscle were harvested 2 hours after the final dose for analysis of pACC/ACC ratios. [1]
For acute effects on hepatic fatty acid synthesis, high-fructose fed db/+ mice were administered MK-3903 orally at single doses of 3, 10, or 30 mg/kg. Hepatic fatty acid synthesis was measured 2 and 8 hours after dosing. [1]
For chronic insulin sensitization studies, DIO mice were treated with MK-3903 orally at doses of 3, 10, or 30 mg/kg BID, or 30 mg/kg once daily (QD), for 12 days. Following a 4-hour fast on day 12, an oral glucose tolerance test (OGTT) was performed. Blood glucose and insulin levels were measured 20 minutes after glucose administration to calculate an insulin resistance index (IRI). [1]
Pharmacokinetic studies were conducted in C57BL/6 mice, Sprague-Dawley rats, and beagle dogs. For mice, MK-3903 was administered intravenously at 2 mg/kg and orally at 10 mg/kg. Blood samples were collected at various time points to determine plasma concentration. [1]

In C57BL/6 mice, MK-3903 exhibited moderate plasma clearance (CLp = 6.8 mL/min/kg), a volume of distribution at steady state (Vdss) of 0.7 L/kg, and a terminal half-life (t1/2) of 2.1 hours. Oral bioavailability (F) was 8.4% with a Cmax of 2.0 µM at 1 hour (tmax). [1]
In Sprague-Dawley rats, CLp was 5.0 mL/min/kg, Vdss was 0.6 L/kg, t1/2 was 1.7 hours, and oral F was 25% (4 mg/kg dose, Cmax = 4.8 µM, tmax = 0.5 hr). [1]
In beagle dogs, CLp was 13 mL/min/kg, Vdss was 1.1 L/kg, t1/2 was 2.4 hours, and oral F was 78% (1 mg/kg dose, Cmax = 1.1 µM, tmax = 0.5 hr). [1]
An earlier analog, compound 36, had high plasma clearance (60 mL/min/kg) and low oral bioavailability (7%) in wild-type mice. In BCRP (ABCG2) knock-out mice, clearance was markedly reduced (~30-fold to 2.9 mL/min/kg) and bioavailability increased dramatically (~10-fold to 77%), identifying it as a BCRP substrate. [1]

MK-3903 is a weak reversible inhibitor of CYP3A4 and CYP2D6 (apparent IC₅₀ > 50 µM) and does not cause time-dependent inhibition of CYP3A4. [1]
It shows moderate binding to the prostanoid DP2 receptor (IC₅₀ = 1.8 µM) but this binding is negligible in the presence of serum. [1]
It inhibited p38-regulated/activated protein kinase (PRAK) with IC₅₀ < 10 µM in a kinase panel screen. [1]
No overt toxicity or lethality was reported in the described mouse studies at the tested doses (up to 30 mg/kg BID for 15 days). A modest body weight loss (3.1%) was observed at the highest dosing frequency (30 mg/kg BID). [1]

[1]. Hit-to-Lead Optimization and Discovery of 5-((5-([1,1'-Biphenyl]-4-yl)-6-chloro-1H-benzo[d]imidazol-2-yl)oxy)-2-methylbenzoic Acid (MK-3903): A Novel Class of Benzimidazole-Based Activators of AMP-Activated Protein Kinase. J Med C

MK-3903 was discovered through a hit-to-lead optimization campaign starting from a benzimidazole-based fragment hit (compound 4, EC₅₀ = 36 µM) identified from a biochemical informer library screen. Key optimizations included introducing a thioaryl acid linker at the 2-position and a hydrophobic biphenyl group at the 5-position of the benzimidazole core, followed by replacing sulfur with oxygen in the linker, which collectively improved potency by ~4 orders of magnitude. [1]
AMPK activation is anticipated to improve metabolic parameters in type 2 diabetes by inhibiting hepatic fatty acid synthesis and gluconeogenesis while stimulating fatty acid oxidation and glucose uptake in muscle. [1]
Based on promising preclinical results showing target engagement, improved lipid metabolism, and insulin sensitization in mouse models, MK-3903 was designated as a development candidate for further research on this class of AMPK activators. [1]

C27H19CLN2O3

454.904365777969

454.108

C, 71.29; H, 4.21; Cl, 7.79; N, 6.16; O, 10.55

1219737-12-8

45256689

White to off-white solid powder

7.1

75.2

2

4

5

33

664

0

FIKQZQDYGXAUHC-UHFFFAOYSA-N

InChI=1S/C27H19ClN2O3/c1-16-7-12-20(13-21(16)26(31)32)33-27-29-24-14-22(23(28)15-25(24)30-27)19-10-8-18(9-11-19)17-5-3-2-4-6-17/h2-15H,1H3,(H,29,30)(H,31,32)

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ≥90 mg/mL
Water: <1mg/mL
Ethanol: <1mg/mL

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.50 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (5.50 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
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6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。