KM 11060

CFTR/cystic fibrosis transmembrane conductance regulator

像 KM11060 这样的小分子校正剂的药理学工具可能有助于研究 F508del-CFTR 加工问题和开发囊性纤维化的治疗方法。在天然上皮组织和培养细胞中，KM11060 都能恢复 F508del-CFTR 运输。 F508del-CFTR 处理在很大程度上被 KM11060 纠正，这还将表面表达提高到低温培养细胞中所见的 75%。在用 KM11060 处理的细胞中，高达 50% 的 F508del-CFTR 被复合糖基化，表明高尔基体转运。 KM11060 是一种对于推进 CF 治疗具有潜在用途的物质。 [1]

当LPS引起急性肺部炎症时，通过阻断PSGL-1（P-选择素糖蛋白配体-1）或P-选择素、用WEB2086阻断PAF或纠正突变的CFTR，F508del小鼠血浆脂氧素A4水平相对于野生型小鼠可显着升高通过 KM11060 进行贩运。 [2]

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CFTR（囊性纤维化跨膜电导调节因子）由中性粒细胞和血小板共同表达。缺乏功能性CFTR可能导致严重的肺部感染和炎症。在这里，我们发现，在急性大肠杆菌肺炎期间，小鼠CFTR（F508del）的突变或CFTR的抑制导致更严重的血小板减少症、肺泡中性粒细胞增多症和细菌病，BAL（支气管肺泡灌洗）中的脂氧素A4/MIP-2（巨噬细胞抑制蛋白-2）或脂氧素A4/中性粒细胞比率降低。在体外，抑制CFTR可促进LPS刺激的中性粒细胞中MIP-2的产生；然而，脂蛋白A4可以剂量依赖性地抑制这种作用。在LPS诱导的急性肺部炎症中，阻断PSGL-1（P-选择素糖蛋白配体-1）或P-选择素，WEB2086拮抗PAF，或KM11060纠正突变的CFTR运输，都可以显著增加F508del中与野生型小鼠相关的血浆脂蛋白A4水平。同时，在LPS攻击下，F508del小鼠的血浆血小板活化因子（PAF）水平和PAF-AH活性高于野生型。与赋形剂治疗的F508del组相比，特异性PAF-AH（PAF乙酰水解酶）抑制剂MAFP抑制PAF的水解可能会加重LPS诱导的F508del小鼠的肺部炎症。特别是，与对照组F508del小鼠相比，F508del鼠血小板耗竭可显著降低血浆脂蛋白A4和PAF-AH活性，并加重LPS诱导的肺部炎症。综上所述，脂氧蛋白A4和PAF参与了大肠杆菌或LPS诱导的CFTR缺陷小鼠的肺部炎症，表明脂氧蛋白A4和PAF可能是改善CFTR缺陷恶化肺部炎症的治疗靶点。[2]

KM11060 是 F508del-CFTR（囊性纤维化跨膜电导调节因子）运输缺陷的新型校正剂。它纠正 F508del-CFTR 运输，增加质膜上功能性 CFTR 的数量 (~75%) 并抑制 PDE5 活性。

KM11060 等小分子校正剂可作为 F508del-CFTR 加工缺陷研究和囊性纤维化治疗药物开发中有用的药理学工具。 KM11060 拯救培养细胞和天然上皮组织中的 F508del-CFTR 运输。 KM11060 部分纠正了 F508del-CFTR 处理，并将表面表达增加至低温培养细胞中观察到的表达水平的 75%。在用 KM11060 处理的细胞中，高达 50% 的 F508del-CFTR 被复合糖基化，表明通过了高尔基体。 KM11060 是一种有前途的化合物，可用于进一步开发 CF 疗法。

Administration of CFTR Inhibitors[2]
Mice were intraperitoneally injected (ip) with MalH-2 (dissolved in PBS, 3 mg/kg) or CFTRinh-172 (dissolved in DMSO, 3 mg/kg) 15∼20 min before intratracheal challenge with E. coli or LPS to establish lung inflammation mouse models.

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E. coli Pneumonia and LPS-induced Acute Lung Inflammation Models[2]
Eight to ten-week old CD1 wild-type and CF mice (targeted F508del gene replacement, obtained from Professor A. Verkman, University of California San Francisco) were used for these studies. Anesthesia was induced with an ip injection of a mixture of ketamine (90 mg/kg) and xylazine (10 mg/kg). [2]
A previously developed direct visualization instillation (DVI) method was used to instill LPS into the airspaces of the lung. The LPS dosage (5 mg/kg) was selected aiming to induce a robust lung inflammation and injury at 24 h as previously reported and no mice died at this dosage. For establishing E. coli pneumonia, 107 cfu of E. coli were instilled into the airspaces of the lung as reported before. E. coli pneumonia and LPS-induced acute lung inflammation mouse models were followed for 4 and 24 respectively. Vital signs of each mouse were observed timely. At the end of experiment, mice were first anesthetized and then sacrificed by cervical dislocation.

[1]. Structural analog of sildenafil identified as a novel corrector of the F508del-CFTR trafficking defect. Mol Pharmacol. 2008 Feb;73(2):478-89.

[2]. Lipoxin A4 and platelet activating factor are involved in E. coli or LPS-induced lung inflammation in CFTR-deficient mice. PLoS One. 2014 Mar 26;9(3):e93003.

The F508del mutation impairs trafficking of the cystic fibrosis transmembrane conductance regulator (CFTR) to the plasma membrane and results in a partially functional chloride channel that is retained in the endoplasmic reticulum and degraded. We recently used a novel high-throughput screening (HTS) assay to identify small-molecule correctors of F508del CFTR trafficking and found several classes of hits in a screen of 2000 compounds (Carlile et al., 2007). In the present study, we have extended the screen to 42,000 compounds and confirmed sildenafil as a corrector using this assay. We evaluated structural analogs of sildenafil and found that one such molecule called KM11060 (7-chloro-4-{4-[(4-chlorophenyl) sulfonyl] piperazino}quinoline) was surprisingly potent. It partially restored F508del trafficking and increased maturation significantly when baby hamster kidney (BHK) cells were treated with 10 nM for 24 h or 10 muM for 2 h. Partial correction was confirmed by the appearance of mature CFTR in Western blots and by using halide flux, patch-clamp, and short-circuit current measurements in unpolarized BHK cells, monolayers of human airway epithelial cells (CFBE41o(-)), and intestines isolated from F508del-CFTR mice (Cftr(tm1Eur)) treated ex vivo. Small-molecule correctors such as KM11060 may serve as useful pharmacological tools in studies of the F508del-CFTR processing defect and in the development of cystic fibrosis therapeutics.[1]

C19H17CL2N3O2S

422.33

421.041

C, 54.04; H, 4.06; Cl, 16.79; N, 9.95; O, 7.58; S, 7.59

774549-97-2

1241327

White to off-white solid powder

1.5±0.1 g/cm3

607.3±65.0 °C at 760 mmHg

321.1±34.3 °C

0.0±1.7 mmHg at 25°C

1.676

4.19

61.89

0

5

3

27

599

0

ClC1C([H])=C([H])C2C(C=1[H])=NC([H])=C([H])C=2N1C([H])([H])C([H])([H])N(C([H])([H])C1([H])[H])S(C1C([H])=C([H])C(=C([H])C=1[H])Cl)(=O)=O

GIEHIZKCIZLXLF-UHFFFAOYSA-N

InChI=1S/C19H17Cl2N3O2S/c20-14-1-4-16(5-2-14)27(25,26)24-11-9-23(10-12-24)19-7-8-22-18-13-15(21)3-6-17(18)19/h1-8,13H,9-12H2

7-chloro-4-(4-((4-chlorophenyl)sulfonyl)piperazin-1-yl)quinoline

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: ~84 mg/mL ( 198.89 mM）
Water: Insoluble
Ethanol: <2 mg/mL

配方 1 中的溶解度: ≥ 5 mg/mL (11.84 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 50.0 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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配方 2 中的溶解度: 10%DMSO+90%corn oil：≥ 5 mg/mL (11.84 mM); 澄清溶液

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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。