FLAP/5-lipoxygenase-activating protein (IC50 = 1.6 nM)

SB203580 和 Quiflaponodium (MK591) 在暴露于 SEB 时可防止人 PBMC 细胞增殖。 SEB 增加的三个基因——组织蛋白酶 L、IL-17 和鸟苷酸结合蛋白 (GBP)-2——被 quilaponodium (MK591) 下调[1]。用 quiflapon 钠 (MK591) 治疗可在数小时内引起细胞凋亡。此外，quilaponodium 还会导致应激激酶 c-Jun N 末端激酶 (JNK) 快速激活。 JNK 对细胞凋亡过程至关重要。当前列腺癌细胞接触奎拉朋钠时会发生凋亡，但 PI3K-Akt 或 ERK 不会受到抑制。此外，Quifflapon 钠和 LY294002 协同作用，导致前列腺癌细胞凋亡 [2]。 Sp1 不受 quilapon 钠 (MK591) 的影响，但 cAMP 反应元件结合蛋白却受影响[4]。

与未治疗的高氧组相比，给予 Quiflapon 钠 (MK591) (20、40 mg/kg) 的高氧组小鼠表现出异常的肺泡化，而治疗组的肺泡化与室内空气对照组相似。然而，不存在炎症[3]。当比较安慰剂和 Quiflapon 钠 (MK591) (320 mg/kg) 治疗组的 Aβ 免疫阳性面积时，治疗组小鼠的淀粉样蛋白负荷在统计学上显着降低。此外，quilapon 钠可显着降低大脑中 IL-1β 的水平。在给予 Quiflapon 钠的小鼠中，总体 CREB 及其 Ser133 磷酸化版本的稳态水平在统计学上显着降低 [4]。

MK591是一种特异性抑制5-Lox活性的合成化合物，目前正在开发用于治疗哮喘。我们观察到用MK591治疗的人前列腺癌细胞在治疗数小时内发生凋亡。细胞凋亡包括严重的形态学改变、磷脂酰丝氨酸外化、PARP的裂解和染色质dna的降解。MK591还诱导应激激酶c-Jun n -末端激酶(JNK)的快速激活，该激酶在细胞凋亡过程中起重要作用。磷脂酰肌醇3′-激酶-Akt/蛋白激酶B (PI3K/Akt)轴是一个众所周知的促生存通路，它通过明确的抗凋亡机制在多种癌细胞中阻止细胞凋亡。有趣的是，我们观察到MK591在不抑制PI3K-Akt或ERK的情况下触发前列腺癌细胞凋亡。此外，MK591与PI3K抑制剂LY294002在诱导前列腺癌细胞凋亡方面具有协同作用。总之，这些发现表明5-Lox抑制诱导的前列腺癌细胞凋亡在不抑制PI3K-Akt或ERK的情况下发生，并表明在这些癌细胞中存在不依赖Akt和ERK的生存机制，通过5-Lox代谢物产生的信号维持生存。[2]

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一系列已完成的抑制剂研究发现了一种特异性的5-LO抑制剂(MK591)，它能够阻断JNK、MAPK、p38kinase和5-LO信号级联，并显著降低促炎细胞因子tnf - α的活性。利用pbmc细胞增殖试验、人近端小管细胞和体内研究(猴子)对MK591进行进一步评估，发现细胞增殖减少。通过利用一组SEB特异性基因，MK591的抑制作用在遗传水平上得到了再次证实。信号活动、抑制剂研究、细胞分析和基因表达分析一致地说明了通路互连物如5-LO以及抑制这种互连物(使用MK591)在SEB诱导的人pbmc中的重要性。[1]

在每个实验中，等量的细胞被镀在六孔板中;24 h后取出培养液，加入含有MK-591 (1 μM, 10 μM或25 μM)或载体的新鲜培养液。孵育24小时后，收集上清液用于测定Aβ和LDH，并在裂解缓冲液中收集细胞颗粒用于免疫印迹分析，如前几段所述。 在转染研究中，使用Lipofectamine 2000 (Invitrogen)将1 μg myc标记的mΔE-Notch-1互补DNA转染N2A-APPswe细胞过夜。除去培养基，加入含有MK-591、L685,458或车辆的新鲜培养基。孵育24小时后，收集细胞裂解液，用western blot分析NICD表达水平。[4]

Starting at 7 months of age, mice were randomized to receive MK-591 (40 mg/kg weight) (n = 11) or vehicle (n = 9) in their chow diet for 8 months until they were 15 months old. Considering that each mouse eats on average 5 g/day of chow diet and the diet is formulated for 320 mg MK-591 per kg diet (Harlan Teklad, WI, USA), the final dose of the active drug was approximately 40 mg/kg weight/day. During the study, mice in both groups gained weight regularly, and no significant difference in weight was detected between the two groups. No macroscopic effect on the overall general health was observed in the animals receiving the active treatment. Post-mortem examination showed no sign of macroscopic pathology in any of the organs considered (spleen, liver, thymus, ileum).[4]

[1]. Effect of 5-lipoxygenase inhibitor MK591 on early molecular and signaling events induced by staphylococcal enterotoxin B in human peripheral blood mononuclear cells. FEBS J. 2008 Jun;275(12):3088-98.

[2]. MK591, a leukotriene biosynthesis inhibitor, induces apoptosis in prostate cancer cells: synergistic action with LY294002, an inhibitor of phosphatidylinositol 3'-kinase. Cancer Lett. 2010 May 28;291(2):167-76.

[3]. 5-Lipoxygenase-activating protein (FLAP) inhibitor MK-0591 prevents aberrant alveolarization in newborn mice exposed to 85% oxygen in a dose- and time-dependent manner. Lung. 2011 Feb;189(1):43-50.

[4]. Involvement of 5-lipoxygenase activating protein in the amyloidotic phenotype of an Alzheimer's disease mouse model. J Neuroinflammation. 2012 Jun 14;9:127.

MK-0591 (3-[1-(4-chlorobenzyl)-3-(t-butylthio)-5-(quinolin-2-yl-methoxy)- indol-2-yl]-2,2-dimethyl propanoic acid, previously L-686,708) is a potent inhibitor of leukotriene (LT) biosynthesis in intact human and elicited rat polymorphonuclear leukocytes (PMNLs) (IC50 values 3.1 and 6.1 nM, respectively) and in human, squirrel monkey, and rat whole blood (IC50 values 510, 69, and 9 nM, respectively). MK-0591 had no effect on rat 5-lipoxygenase. MK-0591 has a high affinity for 5-lipoxygenase activating protein (FLAP) as evidenced by an IC50 value of 1.6 nM in a FLAP binding assay and inhibition of the photoaffinity labelling of FLAP by two different photoaffinity ligands. Inhibition of activation of 5-lipoxygenase was shown through inhibition of the translocation of the enzyme from the cytosol to the membrane in human PMNLs. MK-0591 was a potent inhibitor of LT biosynthesis in vivo, first, following ex vivo challenge of blood obtained from treated rats and squirrel monkeys, second, in a rat pleurisy model, and, third, as monitored by inhibition of the urinary excretion of LTE4 in antigen-challenged allergic sheep. Inhibition of antigen-induced bronchoconstriction by MK-0591 was observed in inbred rats pretreated with methysergide, Ascaris-challenged squirrel monkeys, and Ascaris-challenged sheep (early and late phase response). These results indicate that MK-0591 is a potent inhibitor of LT biosynthesis both in vitro and in vivo indicating that the compound will be suitable for assessing the role of leukotrienes in pathological situations.Can J Physiol Pharmacol. 1992 Jun;70(6):799-807.

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Bronchopulmonary dysplasia is characterized by prolonged oxygen dependency due to compromised gas-exchange capability. This is attributable mainly to inadequate and aberrant alveolarization resulting from insults like hyperoxia. Leukotrienes are associated with hyperoxia-induced inhibition of alveolarization. We hypothesized that a 5-lipoxygenase-activating protein (FLAP) inhibitor given while newborn mice were exposed to 85% oxygen would prevent aberrant alveolarization in a dose- and time-dependent manner. Newborn mice were exposed to either room air or hyperoxia for 14 days. Pups were treated with either vehicle or MK-0591 10, 20, or 40 mg/kg subcutaneously daily for days 1-4, 5-9, or 10-14. On day 14, the lungs were inflated, fixed, and stained for histopathological and morphometric analyses. Hyperoxia groups treated with MK-0591 20 or 40 mg/kg during days P1-P4 or P10-P14 showed alveolarization that resembled that of room air controls while untreated hyperoxia groups showed definite evidence of aberrant alveolarization but no inflammation. In a hyperoxia-exposed newborn mice model, a FLAP inhibitor given during critical window periods may prevent aberration of alveolarization in a dose- and time-dependent manner.Lung. 2011 Feb;189(1):43-50.

C34H34N2O3SCL-.NA+

609.15316

608.188

C, 67.04; H, 5.63; Cl, 5.82; N, 4.60; Na, 3.77; O, 7.88; S, 5.26

147030-01-1

Quiflapon;136668-42-3

23672584

Typically exists as White to off-white solids at room temperature

751.3ºC at 760mmHg

408.2ºC

1.03E-23mmHg at 25°C

7.679

92.48

0

5

10

42

879

0

[O-]C(C(C)(C)CC(N1CC2=CC=C(Cl)C=C2)=C(SC(C)(C)C)C3=C1C=CC(OCC4=NC5=CC=CC=C5C=C4)=C3)=O.[Na+]

YPURUCMVRRNPHJ-UHFFFAOYSA-M

InChI=1S/C34H35ClN2O3S.Na/c1-33(2,3)41-31-27-18-26(40-21-25-15-12-23-8-6-7-9-28(23)36-25)16-17-29(27)37(20-22-10-13-24(35)14-11-22)30(31)19-34(4,5)32(38)39;/h6-18H,19-21H2,1-5H3,(H,38,39);/q;+1/p-1

sodium 3-(3-(tert-butylthio)-1-(4-chlorobenzyl)-5-(quinolin-2-ylmethoxy)-1H-indol-2-yl)-2,2-dimethylpropanoate

MK 0591 sodium; MK-0591; MK0591; L 686708; L-686,708; MK 591; MK591; QUIFLAPON SODIUM; 147030-01-1; MK591; Quiflapon (sodium); MK-591; Quiflapon Sodium [USAN]; MK-591 (sodium); MK 591; MK-591

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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

DMSO : ~50 mg/mL (~82.08 mM)
H2O : < 0.1 mg/mL

配方 1 中的溶解度: ≥ 2.75 mg/mL (4.51 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 27.5 mg/mL澄清DMSO储备液加入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: ≥ 2.75 mg/mL (4.51 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 27.5 mg/mL 澄清 DMSO 储备液加入900 μL 玉米油中，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;
5、为保证最佳实验结果，工作液请现配现用！
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。