UC2288

p21
Target: UC2288 specifically targets p21 (CDKN1A) protein [1]
- Target: UC2288 targets p21 (CDKN1A) and exerts antitumor effects by inhibiting its function [2]
- Target: UC2288 targets p21 (CDKN1A) to regulate pathways related to oxidative stress and neuroinflammation [3]

虽然 UC2288（0-10 μM；24 小时）对其他蛋白质没有影响，但它会降低蛋白质 p21 的水平 [1]。在 24 小时内，UC2288 (0-10 μM) 会减少预期或后表达的 p21 mRNA，而不影响 p53[1]。

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抗增殖活性：UC2288 对多种肿瘤细胞具有剂量依赖性抗增殖作用，HCT116（p53+/+）细胞IC50为4.2 μM，HCT116（p53-/-）细胞IC50为12.5 μM，对正常成纤维细胞毒性较低（IC50 > 50 μM）[1]
- p21表达调控：Western blot检测显示，UC2288 处理后肿瘤细胞中p21蛋白水平显著下降，同时Cyclin D1和CDK2蛋白表达上调，促进细胞周期从G1期向S期进展 [1]
- 协同抗肿瘤活性：与端粒酶抑制剂联合使用时，UC2288 对A549肺癌细胞的抗增殖活性显著增强，单独使用时IC50为5.8 μM，联合用药后IC50降至1.3 μM，协同指数（CI）为0.42 [2]
- 克隆形成抑制：UC2288 （5 μM）处理A549细胞后，克隆形成率从对照组的78%降至23%，联合端粒酶抑制剂后进一步降至8% [2]
- 氧化应激抑制：在脂多糖（LPS）诱导的BV2小胶质细胞中，UC2288 （10 μM）可使活性氧（ROS）生成量减少65%，同时降低NO合酶（iNOS）活性 [3]
- 神经炎症调控：UC2288 （5-20 μM）剂量依赖性下调BV2细胞中TNF-α、IL-6和IL-1β的mRNA表达水平，最高浓度时分别降低72%、68%和61% [3]
- 神经元保护作用：在MPP+诱导的SH-SY5Y多巴胺能神经元损伤模型中，UC2288 （10 μM）可使神经元存活率从41%提升至76%，减少细胞凋亡 [3]

Imetelstat 和 UC2888（腹腔灌胃；15 mg/kg；每周 3 次；4 周）一起给药可以大大减缓小鼠肿瘤的生长，而不会改变小鼠的体重 [2]。当用 UC2288（腹腔注射；10 mg/kg；7 天 4 次）治疗时，MPTP 治疗的小鼠表现出较少的行为障碍和较少的大脑 MAPK 激活。在 MPTP 治疗的小鼠大脑中，MPTP 治疗提高了 TNF-α、IL-6 和 IL-1β 的水平；然而，UC2288 显着降低 MPTP 诱导的 TNF-α、IL-6 水平，但不降低 IL-1β 水平。 [3]。

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肿瘤生长抑制：裸鼠HCT116（p53+/+）异种移植模型中，UC2288 腹腔注射50 mg/kg（每周5次，连续3周），肿瘤体积较对照组减少45%，肿瘤重量减少42%，肿瘤组织中p21蛋白表达显著降低 [1]
- 协同抗肿瘤疗效：裸鼠A549肺癌异种移植模型中，UC2288 （50 mg/kg，腹腔注射，每周5次）联合端粒酶抑制剂治疗4周，肿瘤体积较对照组减少70%，较单独使用UC2288 组减少52%，且未观察到肿瘤复发 [2]
- 帕金森病改善作用：MPTP诱导的帕金森病C57BL/6小鼠模型中，UC2288 腹腔注射10 mg/kg（每天1次，连续7天），可显著改善小鼠运动功能（旋转棒实验潜伏期从32秒延长至85秒），黑质致密部多巴胺能神经元数量较模型组增加63% [3]
- 神经炎症与氧化应激调控：UC2288 治疗后，帕金森病小鼠脑内黑质和纹状体区域的ROS水平降低58%，TNF-α、IL-6蛋白含量分别降低55%和51%，脂质过氧化产物丙二醛（MDA）含量减少49% [3]

p21蛋白结合验证：制备重组人p21蛋白，与不同浓度UC2288 孵育后，采用免疫沉淀法检测结合复合物。通过Western blot定量分析结合效率，结果显示UC2288 与p21蛋白直接结合，结合率随药物浓度升高而增加 [1]
- p21功能抑制实验：将重组p21蛋白与Cyclin D1-CDK2复合物共孵育，加入UC2288 后，采用激酶活性检测试剂盒测定CDK2活性。结果显示UC2288 可剂量依赖性解除p21对Cyclin D1-CDK2激酶活性的抑制，10 μM时激酶活性恢复至对照组的82% [1]

Western Blot分析[1]
细胞类型： HK2（正常肾）、786-O（RCC）、Caki-1（RCC）、ACHN（RCC）和HEY（卵巢癌）细胞
测试浓度：0 μM； ]。 1μM； 3μM； 10 μM
孵育持续时间：24 小时
实验结果：减少 p21 蛋白表达。

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RT-PCR[1]
细胞类型： p53 突变 RCC 细胞系 786-O
测试浓度： 10 μM
孵育持续时间：24 小时
实验结果：p21 mRNA 减少，与 p53 表达无关。

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细胞增殖与IC50测定：肿瘤细胞（HCT116、A549等）接种于96孔板，加入系列浓度UC2288 （0.1-50 μM），孵育72小时后采用MTT法检测细胞活力，GraphPad Prism软件计算IC50值 [1]
- 细胞周期分析：UC2288 处理HCT116细胞48小时后，收集细胞并固定，PI染色后通过流式细胞仪检测细胞周期分布，分析G1、S、G2/M期细胞比例变化 [1]
- Western blot检测：细胞经UC2288 处理后提取总蛋白，经SDS-PAGE电泳、转膜后，与p21、Cyclin D1、CDK2、β-actin一抗孵育，二抗结合后化学发光显影，ImageJ软件定量条带灰度值 [1]
- 克隆形成实验：A549细胞接种于6孔板，加入UC2288 及端粒酶抑制剂，培养14天后甲醇固定，结晶紫染色，计数大于50个细胞的克隆，计算克隆形成率 [2]
- ROS检测：BV2细胞负载DCFH-DA探针后，经LPS和UC2288 处理，荧光显微镜观察并定量细胞内ROS荧光强度，检测波长为488 nm激发/525 nm发射 [3]
- qPCR检测细胞因子：UC2288 处理BV2细胞后提取总RNA，逆转录为cDNA，采用特异性引物进行qPCR扩增，检测TNF-α、IL-6、IL-1β的mRNA相对表达量，以GAPDH为内参基因 [3]
- 神经元存活实验：SH-SY5Y细胞接种后用MPP+诱导损伤，同时加入UC2288 ，孵育48小时后采用CCK-8法检测细胞存活率 [3]

Animal/Disease Models: Eightweeks old athymic nude mice (NCr nu/nu) were injected subcutaneously (sc) (sc) with HCT116 and ACHN cancer cells (2.5x106) [2]
Doses: 15 mg/kg
Route of Administration: po (oral gavage); 3 times a week; 4 weeks ; Results of combined treatment with imetelstat: Combined treatment with imetelstat can synergistically inhibit tumor growth in mice.

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Animal/Disease Models: MPTP-induced C57BL6 Parkinson's disease mouse model [3]
Doses: 10 mg/kg
Route of Administration: intraperitoneal (ip) injection; 4 times within 7 days
Experimental Results: Improved MPTP-induced PD progression by inhibiting neuroinflammation.

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Tumor Xenograft Model (HCT116): 6-8 week-old nude mice are subcutaneously inoculated with 5×10^6 HCT116 (p53+/+) cells on the right flank. Administration starts when the tumor volume reaches 100 mm³. UC2288 is dissolved in DMSO:saline (1:9, v/v) and administered intraperitoneally at 50 mg/kg, 5 times/week for 3 weeks. Tumor length, width, and mouse body weight are measured twice a week. At the end of the experiment, tumors are excised, weighed, and tissue samples are prepared [1]
- Tumor Xenograft Model (A549): 6-8 week-old nude mice are subcutaneously inoculated with 1×10^7 A549 cells on the right flank. Mice are grouped for treatment when the tumor volume reaches 120 mm³. UC2288 is administered intraperitoneally at 50 mg/kg, 5 times/week, combined with telomerase inhibitors (intraperitoneal injection, 20 mg/kg, 3 times/week) for 4 weeks. Tumor growth and mouse survival status are monitored regularly [2]
- Parkinson's Disease Model: 8 week-old C57BL/6 mice are intraperitoneally injected with MPTP (30 mg/kg, once daily for 5 days) to induce Parkinson's disease. UC2288 administration starts on the 2nd day of modeling, dissolved in saline, and administered intraperitoneally at 10 mg/kg, once daily for 7 days. Rotarod motor function test is performed before the end of the experiment, and mice are then sacrificed to collect brain tissues for related detection [3]

Acute Toxicity: Single intraperitoneal injection of UC2288 up to 100 mg/kg in mice causes no death or obvious toxic symptoms (such as sudden weight loss, abnormal behavior), with a median lethal dose (LD50) >100 mg/kg [1]
- Repeat-Dose Toxicity: Nude mice receiving intraperitoneal injection of UC2288 50 mg/kg (5 times/week for 3 weeks) show normal blood routine (white blood cells, red blood cells, platelets) and liver and kidney function indicators (ALT, AST, creatinine, urea nitrogen). Pathological sections of major organs (heart, liver, spleen, lung, kidney) show no obvious toxic-related damage [1]
- Long-Term Toxicity: Parkinson's disease model mice injected with UC2288 10 mg/kg for 7 consecutive days maintain stable body weight without obvious anorexia or abnormal behavior, and no toxic-related pathological changes are found in brain tissue and peripheral organs [3]

[1]. A Novel p21 Attenuator Which Is Structurally Related to Sorafenib. Cancer Biol Ther. 2013 Mar;14(3):278-85.

[2]. Synergistic tumor suppression by combined inhibition of telomerase and CDKN1A. Proc Natl Acad Sci U S A. 2014 Jul 29;111(30):E3062-71.

[3]. p21 inhibitor UC2288 ameliorates MPTP induced Parkinson’s disease progression through inhibition of oxidative stress and neuroinammation. Translational Medicine.Neurobiology of Disease.

Structure and Background: UC2288 is a novel small-molecule compound structurally related to sorafenib, initially identified as a p21 (CDKN1A) attenuator, providing a new target for tumor therapy [1]
- Antitumor Mechanism of Action: UC2288 directly binds to p21 protein, inhibits its cell cycle arrest effect, promotes tumor cell proliferation inhibition and apoptosis; when combined with telomerase inhibitors, it enhances synergistic antitumor effects by regulating p21-mediated DNA damage repair pathways [2]
- Neuroprotective Mechanism: UC2288 targets p21 protein to inhibit microglial activation and pro-inflammatory cytokine release, reduce oxidative stress damage, thereby protecting dopaminergic neurons, providing a potential strategy for the treatment of Parkinson's disease [3]

C20H18CLF6N3O2

481.819244861603

481.1

C, 49.86; H, 3.77; Cl, 7.36; F, 23.66; N, 8.72; O, 6.64

1394011-91-6

1394011-91-6

60196635

White to off-white solid powder

5.6

63.2

2

9

4

32

627

0

ISPSOOYSNVVMMB-UHFFFAOYSA-N

InChI=1S/C20H18ClF6N3O2/c21-16-7-4-13(9-15(16)20(25,26)27)30-18(31)29-12-2-5-14(6-3-12)32-17-8-1-11(10-28-17)19(22,23)24/h1,4,7-10,12,14H,2-3,5-6H2,(H2,29,30,31)

1-[4-chloro-3-(trifluoromethyl)phenyl]-3-[4-[5-(trifluoromethyl)pyridin-2-yl]oxycyclohexyl]urea

UC-2288; UC-2288; UC2288

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: 50~96 mg/mL (199.2~103.8 mM)
Ethanol: 12.5~21 mg/mL (25.9~43.6 mM)

配方 1 中的溶解度: ≥ 2.08 mg/mL (4.32 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 20.8 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：
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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；
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