PLK1 (IC50 = 9 nM); PLK2 (IC50 = 260 nM); PDGFR (IC50 = 18 nM); Src (IC50 = 155 nM); BCR-ABL (IC50 = 32 nM); Cdk1 (IC50 = 260 nM); Flt1 (IC50 = 42 nM); Fyn (IC50 = 182 nM)
Rigosertib, which has an IC50 of 9 nM, is a non-ATP-competitive inhibitor of PLK1. Moreover, rigosertib shows inhibition with an IC50 of 18-260 nM against PLK2, PDGFR, Flt1, BCR-ABL, Fyn, Src, and CDK1. 94 distinct tumor cell lines, including BT27, MCF-7, DU145, PC3, U87, A549, H187, RF1, HCT15, SW480, and KB cells, exhibit cell killing activity in response to rigosertib, with an IC50 of 50–250 nM. Rigosertib, however, has little to no effect in normal cells, such as HFL, PrEC, HMEC, and HUVEC, unless the concentration is higher than 5–10 μM. Rigosertib (100-250 nM) causes spindle abnormalities and apoptosis in HeLa cells.[1] With an IC50 of 50-100 nM, rigorsertib also inhibits a number of multidrug-resistant tumor cell lines, such as MES-SA, MES-SA/DX5a, CEM, and CEM/C2a. Rigosertib (0.25–5 μM) causes an accumulation of cells with subG1 DNA content, initiates apoptotic pathways, and inhibits cell cycle progression in G2/M phase in DU145 cells. Rigosertib (50 nM-0.5 μM) causes caspase 3/7 activation and viability loss in A549 cells.[2] According to a recent study, rigosertib causes CLL (chronic lymphocytic leukemia) cells to undergo apoptosis without harming healthy B-cells or T-cells. Additionally, rigorsertib inhibits the migration of leukemic cells induced by SDF-1 and nullifies the pro-survival effect of follicular dendritic cells on CLL cells.[3]

体外活性：Rigosertib 是 PLK1 的非 ATP 竞争性抑制剂，IC50 为 9 nM。 Rigosertib 还表现出对 PLK2、PDGFR、Flt1、BCR-ABL、Fyn、Src 和 CDK1 的抑制作用，IC50 为 18-260 nM。 Rigosertib 对 94 种不同的肿瘤细胞系显示出细胞杀伤活性，IC50 为 50-250 nM，包括 BT27、MCF-7、DU145、PC3、U87、A549、H187、RF1、HCT15、SW480 和 KB 细胞。而在正常细胞（例如 HFL、PrEC、HMEC 和 HUVEC）中，Rigosertib 的作用很小或没有作用，除非其浓度大于 5-10 μM。在 HeLa 细胞中，Rigosertib (100-250 nM) 会诱导纺锤体异常和细胞凋亡。 Rigosertib 还抑制多种耐药肿瘤细胞系，包括 MES-SA、MES-SA/DX5a、CEM 和 CEM/C2a，IC50 为 50-100 nM。在 DU145 细胞中，Rigosertib (0.25-5 μM) 可阻断 G2/M 期的细胞周期进展，导致含有 subG1 DNA 含量的细胞积累，并激活细胞凋亡途径。在 A549 细胞中，Rigosertib (50 nM-0.5 μM) 会导致活力丧失和 caspase 3/7 激活。在最近的一项研究中，Rigosertib 可诱导慢性淋巴细胞白血病 (CLL) 细胞凋亡，但对 T 细胞或正常 B 细胞没有毒性。 Rigosertib 还消除滤泡树突状细胞对 CLL 细胞的促生存作用，并减少 SDF-1 诱导的白血病细胞迁移。激酶测定：将重组 PLK1 (10 ng) 与不同浓度的 Rigosertib 在 15 µL 反应混合物（50 mM HEPES、10 mM MgCl2、1 mM EDTA、2 mM 二硫苏糖醇、0.01% NP-40 [pH 7.5]）中孵育，室温下 30 分钟。激酶反应在 30 °C 下进行 20 分钟，体积为 20 µL（15 µL 酶 + 抑制剂、2 µL 1 mM ATP）、2 µL γ32P-ATP (40 µCi) 和 1 µL 重组 Cdc25C（100 ng）或酪蛋白（1 μg）底物。在 20 µL 2× Laemmli 缓冲液中煮沸 2 分钟终止反应。磷酸化底物通过 18% SDS-PAGE 分离。将凝胶干燥并暴露于 X 射线胶片 3-10 分钟。细胞测定：细胞（许多肿瘤细胞系，包括 BT20、MCF-7、DU145、PC3、U87、A549、H187、RF1、HCT15、HeLa 和 Raji 细胞）在补充有 10% 的 DMEM 或 RPMI 中生长胎牛血清和1单位/mL青霉素-链霉素溶液。将肿瘤细胞以 1 × 105 个细胞/mL/孔的密度接种到六孔培养皿中，24 小时后添加不同浓度的 Rigosertib。处理 96 小时后，从重复孔中测定细胞计数。活细胞总数通过台盼蓝排除法测定。

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Rigosertib 在所测试的94种不同肿瘤细胞系中均能诱导细胞凋亡，其GI50范围在50至200 nM之间。[1]
在美国国家癌症研究所（NCI）开发治疗计划（DTP）筛选的57种人类癌细胞系面板中，Rigosertib 在纳摩尔浓度下显示出广谱的生长抑制活性，包括对多药耐药（MDR）细胞系。[1]
对多种化疗药物耐药的MDR阳性细胞系MES-SA/DX5和CEM/C2，对Rigosertib 的敏感性与亲本细胞系相同，表明不存在交叉耐药性。[1]
用阿非迪霉素阻滞于G1/S期的HeLa细胞释放后，用Rigosertib (250 nM) 处理，导致免疫沉淀物中Plk1激酶活性在所有细胞周期阶段急剧降低。这种抑制并非由于Plk1蛋白降解或合成减少，因为其稳态水平与对照组相似。[1]
流式细胞术分析显示，Rigosertib 处理导致HeLa细胞在从阿非迪霉素阻滞释放后8至16小时期间积聚在细胞周期的G2/M期，随后细胞活力迅速丧失。[1]
用Rigosertib (250 nM) 处理的HeLa细胞的共聚焦显微镜检查显示有丝分裂纺锤体形成存在严重异常，包括多极纺锤体、染色体排列错乱以及染色体-纺锤体连接不稳定。[1]
Rigosertib 处理的HeLa细胞中γ-微管蛋白的免疫荧光染色显示中心体定位异常和碎片化，存在多个随机分布的成对中心粒。[1]
膜联蛋白V/PI染色和流式细胞术表明，Rigosertib 处理可诱导HeLa细胞凋亡，细胞在24小时内变为膜联蛋白V阳性，并在48小时内变为双阳性（膜联蛋白V/PI）。[1]
Western印迹分析证实，Rigosertib 处理的HeLa细胞中PARP在24小时即发生切割，48小时时变得非常明显。Caspase-3活性也显著上调。[1]
几种正常人细胞系（成纤维细胞HFL、前列腺上皮细胞PrEC、乳腺上皮细胞HMEC、脐静脉内皮细胞HUVEC）对Rigosertib 的凋亡效应具有高度抗性，仅在5–10 µM浓度下才发生细胞死亡。流式细胞术显示Rigosertib 处理的HFL细胞周期进展正常。[1]
在微管蛋白聚合实验中，Rigosertib (5 µM) 不抑制纯化的牛脑微管蛋白的体外聚合，这与紫杉醇（稳定微管）和长春碱（抑制聚合）不同。这表明该化合物不直接与β-微管蛋白结合以破坏微管动力学。[1]

在 Bel-7402、MCF-7 和 MIA-PaCa 细胞的小鼠异种移植模型中，Rigosertib (250 mg/kg) 显着抑制肿瘤生长。 Rigosertib (200 mg/kg) 在 BT20 细胞的小鼠异种移植模型中显示出对肿瘤生长的抑制作用。

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为了确定体内疗效，我们利用了裸鼠模型系统。将高度侵袭性的人雌激素阴性乳腺癌细胞系（BT20）异种移植到无胸腺裸鼠体内。采用Q2D×20方案，用200mg/kg的28（Rigosertib）治疗动物。当肿瘤大小约为70mm3时，对动物进行治疗。图5A显示，腹腔内注射Rigosertib能够显著抑制肿瘤的生长。在22天的时间里，载体治疗小鼠的肿瘤体积平均增加了5倍（70 mm3-480 mm3），而Q2D Rigosertib治疗的肿瘤体积仅增加了2.5倍（70 mm 3-180 mm3）。根据体重和身体观察结果，Rigosertib在这些剂量下耐受良好（图5B）。这些研究表明，Rigosertib对人类肿瘤异种移植物有效，同时在本研究测试的时间表中没有显示出毒性迹象。[1]
在体内，Rigosertib没有表现出血液毒性、肝损伤或神经毒性，并且是各种异种移植物裸鼠模型中肿瘤生长的强效抑制剂。Rigosertib/ON01910与几种化疗药物显示出很强的协同作用，通常会导致肿瘤完全消退[1]。

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在携带皮下Bel-7402（人肝癌）异种移植瘤的雌性无胸腺裸鼠中，腹腔注射Rigosertib（250 mg/kg，隔日给药）可有效抑制肿瘤生长。与奥沙利铂（100 mg/kg）联用可导致肿瘤块完全消失。根据稳定的体重判断，未观察到毒性迹象。[1]
在携带MCF-7（人乳腺癌）异种移植瘤的裸鼠中，腹腔注射Rigosertib（250 mg/kg，隔日给药）可抑制肿瘤生长。与多柔比星（2.5 mg/kg）联用可导致肿瘤完全缓解。体重保持稳定。[1]
在携带MIA-PaCa（人胰腺癌）异种移植瘤的裸鼠中，腹腔注射Rigosertib（250 mg/kg，隔日给药）可抑制肿瘤生长。与吉西他滨（100 mg/kg）联用可导致肿瘤块显著减小。体重保持稳定。[1]
对来自Rigosertib 治疗小鼠的肿瘤裂解液分析显示，Plk1激酶活性极少或没有，而CDK1活性降低但未完全消除。Western印迹分析证实Plk1和Cyclin B1蛋白的稳态水平与对照组相似。[1]

将重组 PLK1 (10 ng) 与不同浓度的 rigosertib 在 15 µL 反应混合物（50 mM HEPES、10 mM MgCl₂、1 mM EDTA、2 mM 二硫苏糖醇）中在室温下孵育 30 分钟，0.01% NP-40 [pH 7.5]）。 20 µL（15 µL 酶 + 抑制剂、2 µL 1 mM ATP）、2 µL γ³²P-ATP (40 µCi) 和 1 µL 重组 Cdc25C (100 ng) 或酪蛋白 (1 μg）底物用于激酶反应，在 30°C 下进行 20 分钟。在 20 µL 2× Laemmli 缓冲液中煮沸 2 分钟，结束反应。 18% SDS-PAGE 用于分离磷酸化底物。干燥后，将凝胶暴露在 X 射线胶片下三到十分钟。

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对于Plk1抑制实验，将10 ng重组Plk1与不同浓度的Rigosertib 或DMSO（对照）在反应缓冲液（50 mM HEPES, 10 mM MgCl2, 1 mM EDTA, 2 mM DTT, 0.01% NP-40，pH 7.5）中混合，并在室温下孵育30分钟。然后通过添加含有γ-[32P]-ATP和重组Cdc25C或酪蛋白底物的反应混合物来启动激酶反应，并在30°C下孵育20分钟。通过煮沸Laemmli缓冲液终止反应，磷酸化的底物通过SDS-PAGE分离，然后进行放射自显影。[1]
对于定量IC50测定，激酶实验操作如上。孵育后，将反应混合物等分点样到P81磷酸纤维素纸上。滤纸用磷酸和丙酮洗涤，并使用液体闪烁计数器测量掺入的放射性。绘制剂量反应曲线以计算IC50值。[1]
进行稳态动力学分析以确定抑制机制。将重组Plk1与Rigosertib 孵育，并在不同浓度ATP或底物（Cdc25C）存在下进行激酶实验。测量反应速度，并使用米氏方程分析数据以确定Km和Vmax。分析表明，增加ATP浓度不改变IC50，而增加底物浓度会增加表现Km而不影响Vmax，这是相对于底物的竞争性抑制的特征。[1]
为了评估细胞来源的Plk1活性，裂解处于有丝分裂期的同步化HeLa细胞。使用特异性抗体从澄清的裂解液中免疫沉淀Plk1。洗涤免疫沉淀物，并在存在或不存在Rigosertib 的情况下，以Cdc25C或酪蛋白为底物进行激酶实验。[1]

通过流式细胞术对用膜联蛋白V和CD19染色的细胞进行体外测定，确定了Rigosertib（ON 01910.Na）对34名患者CLL细胞的细胞毒性。使用Affymetrix微阵列、流式细胞术、蛋白质印迹和与基质细胞共培养的全球基因表达谱来描绘on 01910.Na的作用机制。[3]

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细胞毒性试验[2]
研究人员使用剂量反应终点测定系统测试了许多肿瘤细胞系。细胞在添加了10%胎牛血清和1单位/mL青霉素-链霉素溶液的DMEM或RPMI中生长。将肿瘤细胞以1.0×105个细胞/mL/孔的细胞密度铺入6孔培养皿中，24小时后加入不同浓度的化合物。处理96小时后，从重复孔中测定细胞计数。通过台盼蓝排斥法测定活细胞总数。 流式细胞术人前列腺肿瘤细胞DU145细胞和正常二倍体人肺成纤维细胞HFL-1细胞在添加了10%胎牛血清和1单位/mL青霉素-链霉素的DMEM中生长。将细胞以1.0×106个细胞/皿的细胞密度铺在100mm2的培养皿上，24小时后，用2.5μM的化合物处理。处理后24小时收获细胞。通过胰蛋白酶消化将细胞从培养皿中取出，并与培养基中发现的未附着细胞结合。将细胞沉淀在磷酸盐缓冲盐水（PBS）中洗涤，并在冰冷的70%乙醇中固定至少24小时。然后用室温PBS洗涤固定的细胞，并在37°C下用碘化丙啶（50mg/mL）和RNase A（0.5mg）染色30分钟。然后在Becton Dickinson流式细胞仪上分析染色细胞，并通过细胞周期分析软件分析数据。

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PARP Wester Blot[2]
DU145和HFL-1细胞以每150mm2板3.0×106个细胞的密度铺板，24小时后用DMSO或28处理。处理48小时后收集细胞，冷冻细胞颗粒。冷冻细胞颗粒在含有蛋白酶抑制剂的1%NP40/PBS裂解缓冲液中裂解。然后在10%-SDS聚丙烯酰胺凝胶上溶解等量的总细胞蛋白。将凝胶转移到硝化纤维纸（S/S）上，与抗PARP抗体（BD）杂交，并使用ECL溶液显影。

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细胞活力和半胱氨酸天冬氨酸蛋白酶3/7活性[2]
将指数增长的A549细胞以3600个细胞/孔的密度接种在100μl含有10%FBS和1%Pen/Strep的DMEM中的白色壁96孔板中。然后让细胞在37°C的培养箱中粘附过夜。第二天，用不同浓度的28或DMSO处理细胞，然后将其放回培养箱中。24小时后，从培养箱中取出培养板，按照制造商的说明，将20μL CellTiter Blue®试剂单独添加到每个孔中。将板缓慢摇动0.5分钟，然后放回培养箱中。3小时后，使用Glomax 96孔板读数器读取荧光。接下来，按照制造商的说明向每个孔中加入120μL Caspase-Glo®3/7试剂。将板缓慢摇动0.5分钟，并在室温下显影2小时。在此期间结束时，使用Glomax 96孔板读数器读取发光。

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一天后，将不同浓度的 Rigosertib 添加到含有 1×10⁵ 细胞/mL/孔的已铺板肿瘤细胞的六孔培养皿中。处理96小时后，从重复的孔中获得细胞计数。通过使用台盼蓝排除法，可以找到活细胞的总数。

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通过将细胞与Rigosertib 孵育96小时进行细胞活力实验，并通过台盼蓝排除法确定活细胞数量。根据剂量反应曲线计算GI50/IC50值。[1]
对于细胞周期分析，使用阿非迪霉素（1 µg/mL，14小时）将HeLa细胞同步在G1/S边界。解除阻滞后，将细胞在有或无Rigosertib 存在下孵育。在指定时间点收集细胞，用乙醇固定，用碘化丙啶和RNase A染色，并通过流式细胞术分析DNA含量。[1]
根据制造商的方案，用FITC偶联的膜联蛋白V和碘化丙啶对细胞进行染色，然后进行流式细胞术分析，以评估细胞凋亡。[1]
通过Western印迹分析PARP切割。将来自处理细胞的总细胞蛋白等量通过SDS-PAGE分离，转移到膜上，并用抗PARP抗体探测以检测切割片段。[1]
使用商业检测试剂盒测量Caspase-3活性。将细胞裂解液与Caspase-3底物Ac-DEVD-pNA孵育，并通过分光光度法在405 nm处测量酶解导致的pNA释放。[1]
对于纺锤体和中心体形态的共聚焦显微镜分析，将生长在盖玻片上的HeLa细胞用Rigosertib 处理，用多聚甲醛固定，透化，并用FITC偶联的单克隆抗α-微管蛋白或抗γ-微管蛋白抗体染色，分别显示微管和中心体。用碘化丙啶对染色体DNA进行复染。使用共聚焦显微镜分析染色的细胞。[1]

Bel-7402 tumor models: subcutaneous injection of 1 × 10⁷ Bel-7402 tumor cells is given to twenty female athymic (NCR-nu/nu) nude mice. Ten to fourteen days later, when the tumor volumes reach 200–250 mm, the mice are divided into four groups, each of which has tumors of the same volume. Alternatively, NSC 266046 (100 mg/kg) and rigosertib (ON01910, 250 mg/kg) can be given intraperitonially on different days. Using traceable digital vernier calipers, tumor measurements are performed twice a week. During every measurement, body weight is calculated. Signs of toxicity in the animals are monitored.

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Nude mouse assay [2]
Female athymic (NCR-nu/nu, Taconic) nude mice were injected with 0.5–1.0 x107 BT20 cells subcutaneously in the hind leg using a 1 mL tuberculin syringe equipped with a 271/2 gauge needle. Approximately 14 days later, mice were paired (N=9) and injected with 200 mg/Kg 28 or Phosphate buffered saline as the vehicle control. The intravenous injections were performed in the mouse tail vein using a 1 mL tuberculin syringe equipped with a 30 gauge needle. The animals were injected following a Q2D X 20 schedule. Tumor measurements (two dimensions) were done three times per week using traceable digital vernier calipers. Tumor volume was calculated using the following equation: V= (Lx(S2)p/6, where L is the longer and S is the shorter of the two dimensions. Body weight was determined during each measurement. The animals were observed for signs of toxicity. The time of tumor volume doubling was calculated and the T-C value (difference in the average times post treatment for tumors of the treated groups to attain a doubling in volume compared to the average of the control group) was determined. We did not observe body weight loss of more than 10% in any group nor were there any animal deaths.

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For the Bel-7402 tumor model, female athymic (NCR-nu/nu) nude mice were injected subcutaneously with 1 × 10^7 Bel-7402 human liver cancer cells. After 10-14 days, when tumor volumes reached 200–250 mm³, mice were divided into groups with similar average tumor volumes. Rigosertib (250 mg/kg) dissolved in PBS was administered intraperitoneally alone or in combination with oxaliplatin (100 mg/kg) on alternate days. Tumor dimensions were measured twice weekly using calipers, and body weight was monitored. [1]
For the MCF-7 tumor model, female athymic nude mice were implanted with a 1 mg estradiol pellet subcutaneously 24 hours before tumor inoculation. Mice were then injected with 1 × 10^7 MCF-7 human breast cancer cells subcutaneously. After tumors reached 200–250 mm³ (10-15 days), mice were divided into groups. Rigosertib (250 mg/kg in PBS) was administered intraperitoneally alone or in combination with doxorubicin (2.5 mg/kg) on alternate days. Tumor size and body weight were monitored. [1]
For the MIA-PaCa tumor model, female athymic nude mice were injected subcutaneously with 1 × 10^7 MIA-PaCa human pancreatic cancer cells. After tumors reached 200–250 mm³ (10-15 days), mice were divided into groups. Rigosertib (250 mg/kg in PBS) was administered intraperitoneally alone or in combination with gemcitabine (100 mg/kg) on alternate days. Tumor size and body weight were monitored. Tumor volume was calculated using the formula: V = L × S² × π/6, where L is the longer and S is the shorter of the two perpendicular dimensions. [1]
At the end of the studies, tumors were excised from selected mice, homogenized in modified RIPA buffer, and clarified by centrifugation. The supernatants were used for immunoprecipitation and kinase assays to assess Plk1 and CDK1 activity, as well as for Western blot analysis. [1]

Rigosertib (compound 28) exhibits good pharmacokinetic and pharmacodynamic properties [2]
It has high water solubility (28 mg/mL), suitable for intravenous, intraperitoneal, and oral administration [2]

In vivo, this compound did not exhibit hematotoxicity, liver damage, or neurotoxicity, and was a potent inhibitor of tumor growth in a variety of xenograft nude mouse models. ON01910 showed strong synergy with several chemotherapeutic agents, often inducing complete regression of tumors.

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In rats, single intravenous bolus doses of 300 and 600 mg/m² of Rigosertib produced no toxicity. A dose of 1200 mg/m² caused only slight toxicity (anogenital staining). The severely toxic dose (STD/LD10) was below 3000 mg/m² (9 of 11 animals died). In 28-day repeat dosing studies, fixed daily doses of 180 mg/m² and 450 mg/m² were tolerated. A dose of 900 mg/m² given twice per week for 4 weeks was well tolerated. [1]
In dogs, single intravenous doses of 2000 and 4000 mg/m² produced gastrointestinal effects (diarrhea, flatulence), with other signs of discomfort at 4000 mg/m². The highest non-severely toxic dose (HNSTD) for a single dose was 2000 mg/m². In 7-day repeat intravenous dosing, 1000 mg/m² daily was well tolerated. In 28-day repeat dosing, daily intravenous doses of 200 and 500 mg/m² were well tolerated. The HNSTD for the 28-day schedule was estimated at 1500-2000 mg/m². Dogs treated at 1000 mg/m² twice per week for 4 weeks tolerated the compound well. [1]
There was no evidence of significant myelotoxicity, neuropathy, or cardiotoxicity in these toxicology studies in rats and dogs. [1]
In nude mouse xenograft efficacy studies, administration of Rigosertib at 250 mg/kg on alternate days did not cause body weight loss or overt signs of toxicity. Bone marrow colony assays from treated animals showed no decrease in colony formation. [1]

[1]. Cancer Cell . 2005 Mar;7(3):275-86.

[2]. J Med Chem . 2011 Sep 22;54(18):6254-76.

[3]. Clin Cancer Res . 2012 Apr 1;18(7):1979-91.

Rigosertib is an N-[2-methoxy-5-({[2-(2,4,6-trimethoxyphenyl)ethenyl]sulfonyl}methyl)phenyl]glycine in which the double bond has E-configuration. It is a non-ATP-competitive inhibitor of PLK1 with an IC50 of 9 nM and exhibits anti-cancer properties. It has a role as a microtubule-destabilising agent, an EC 2.7.11.21 (polo kinase) inhibitor, an apoptosis inducer and an antineoplastic agent. It is a conjugate acid of a rigosertib(1-).
Rigosertib has been used in trials studying the treatment and basic science of MDS, RAEB, Cancer, Hepatoma, and Neoplasms, among others.
Rigosertib is a synthetic benzyl styryl sulfone analogue and Ras mimetic, with potential antineoplastic activity. Upon administration, rigosertib targets and binds to Ras-binding domain (RBD) found in many Ras effector proteins, including Raf kinase and phosphatidylinositol 3-kinase (PI3K). This prevents Ras from binding to its targets and inhibits Ras-mediated signaling pathways, including Ras/Raf/Erk, Ras/CRAF/polo-like kinase1 (Plk1), and Ras/ PI3K/Akt signaling pathways. This induces cell cycle arrest and apoptosis and inhibits proliferation in a variety of susceptible tumor cells.

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Rigosertib (ON01910) is a novel small molecule inhibitor of Plk1 that is non-competitive with ATP and likely binds at or near the substrate binding domain. [1]
It induces mitotic arrest in a wide variety of human tumor cells, characterized by spindle abnormalities and centrosome fragmentation, leading to apoptotic death. [1]
It shows potent activity against multi-drug resistant (MDR) tumor cell lines, and attempts to select for resistance to Rigosertib in tumor cells were unsuccessful. [1]
It exhibits a high therapeutic index, with potent antitumor activity in vivo and low toxicity in normal cells and animals. Based on this profile, Rigosertib had entered Phase I clinical trials for cancer therapy at the time of the study. [1]

C21H25NO8S

451.4901

451.13

C, 55.87; H, 5.58; N, 3.10; O, 28.35; S, 7.10

592542-59-1

Rigosertib sodium;592542-60-4;(E/Z)-Rigosertib sodium;1225497-78-8

6918736

White to light yellow solid powder

1.332±0.06 g/cm3

172-174 ºC (acetone )

3.957

128.77

2

9

11

31

678

0

O=S(/C=C/C(C(OC)=C1)=C(OC)C=C1OC)(CC2=CC=C(OC)C(NCC(O)=O)=C2)=O

OWBFCJROIKNMGD-BQYQJAHWSA-N

InChI=1S/C21H25NO8S/c1-27-15-10-19(29-3)16(20(11-15)30-4)7-8-31(25,26)13-14-5-6-18(28-2)17(9-14)22-12-21(23)24/h5-11,22H,12-13H2,1-4H3,(H,23,24)/b8-7+

2-[2-methoxy-5-[[(E)-2-(2,4,6-trimethoxyphenyl)ethenyl]sulfonylmethyl]anilino]acetic acid

ON-01910; ON01910; ON 01910; Rigosertib; 592542-59-1; UNII-67DOW7F9GL; ON-01910; Rigosertib [USAN:INN]; 67DOW7F9GL; Rigosertib [USAN];brand name: Estybon

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： (1). 请将本产品存放在密封且受保护的环境中（例如氮气保护），避免吸湿/受潮。  (2). 该产品在溶液状态不稳定，请现配现用。

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

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.54 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 (5.54 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
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900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.5 mg/mL (5.54 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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配方 4 中的溶解度: Saline: 30 mg/mL

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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。