TAS-103 dihydrochloride

Topoisomerase II; Topoisomerase I
DNA Topoisomerase I (Topo I) (IC50 = 0.15 μM, recombinant Topo I-mediated DNA relaxation assay; stabilizes Topo I-DNA cleavable complexes) [1][3]
DNA Topoisomerase II (Topo II) (IC50 = 0.2 μM, recombinant Topo II-mediated DNA decatenation assay; stabilizes Topo II-DNA cleavable complexes) [1][3]
(Note: Dual inhibitor of Topo I and Topo II, with similar potency against both enzymes; no significant inhibition of other DNA-processing enzymes (e.g., DNA polymerase, helicase) at concentrations up to 10 μM) [3]

体外活性：TAS-103 diHClide（也称为 BMS-247615）是 DNA 拓扑异构酶 I/II 的双重抑制剂。它是一种喹啉衍生物，在小鼠和人类肿瘤模型中显示出抗肿瘤活性。 TAS-103 在 CCRF-CEM 细胞上具有活性，IC50 值为 5 nM。 0.1 μM 的 TAS-103 显着增加单个 CCRF-CEM 细胞中拓扑 IIα FITC 免疫荧光的水平。 TAS-103 抑制 HeLa 细胞的活力，IC50 为 40 nM。 10 μM 的 TAS-103 会破坏信号识别颗粒 (SRP) 复合物的形成，并诱导 SRP14 和 SRP19 不稳定并最终降解。尽管TAS-103已被报道是一种有效的拓扑异构酶II毒物。然而，其他研究表明细胞对 TAS-103 的敏感性与拓扑异构酶 II 表达无关。 TAS-103（30 mg/kg，静脉注射）对携带 Lewis 肺癌 (LLC) 细胞的小鼠产生显着的肿瘤生长抑制，且体重没有明显下降，并且脂质体 TAS-103 比游离 TAS-103 活性更高。细胞测定：CCRF-CEM 人急性淋巴细胞白血病细胞在补充有 3 mM L-谷氨酰胺、10% 胎牛血清、50 U/mL 青霉素和 40 μg/mL 链霉素的 RPMI-1640 中于 37°C 培养。含有 5% CO2 的潮湿气氛。 TAS-103、CPT 和 DACA 溶解在 DMSO 中。将呈指数增长的细胞 (∼5 × 105) 暴露于任一药物中 2 小时。药物暴露后，细胞在冷磷酸盐缓冲盐水中离心（400 × g，3 分钟）洗涤两次

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1. 双抑制Topo I/II并稳定切割复合物：TAS-103 dihydrochloride剂量依赖性抑制重组Topo I（IC50=0.15 μM）和Topo II（IC50=0.2 μM）活性。在HL-60人白血病细胞中，它稳定Topo I-DNA和Topo II-DNA切割复合物，表现为线性DNA片段生成增加（中性琼脂糖凝胶电泳）。0.5 μM剂量下，Topo I介导的DNA切割增强3.2倍，Topo II介导的切割增强2.8倍（vs溶媒组）[1][3]
2. 强效抑制癌细胞增殖：TAS-103 dihydrochloride抑制多种人类癌细胞系增殖，包括白血病（HL-60，IC50=0.08 μM；K562，IC50=0.12 μM）、结肠癌（Colon 26，IC50=0.15 μM）和乳腺癌（MCF-7，IC50=0.2 μM）（72小时MTT实验）；对正常人成纤维细胞（WI-38）影响极小（IC50>10 μM）[1][2]
3. 诱导DNA损伤和凋亡：TAS-103 dihydrochloride（0.1-1 μM）剂量依赖性诱导HL-60细胞DNA双链断裂（γ-H2AX焦点形成，免疫荧光）。0.5 μM剂量下，γ-H2AX焦点较溶媒组增加4.5倍；通过内源性途径诱导凋亡：Annexin V-FITC/PI染色显示48小时凋亡率达42%，伴随caspase-3/9激活和PARP切割（Western blot）[1][3]
4. 与经典Topo抑制剂无交叉耐药：TAS-103 dihydrochloride对喜树碱耐药（Topo I突变）和阿霉素耐药（Topo II下调）癌细胞系仍保持抗增殖活性，IC50值与亲本细胞系相近（HL-60/Cpt：IC50=0.1 μM；MCF-7/Dox：IC50=0.25 μM）[3]

TAS-103（30 mg/kg，静脉注射）对携带 Lewis 肺癌 (LLC) 细胞的小鼠产生显着的肿瘤生长抑制，且体重没有明显下降，并且脂质体 TAS-103 比游离 TAS-103 活性更高。

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1. 小鼠异种移植模型肿瘤生长抑制：携带Colon 26结肠癌异种移植瘤的BALB/c裸鼠，静脉注射脂质体TAS-103 dihydrochloride（5 mg/kg、10 mg/kg），每周一次，连续4周。10 mg/kg组平均肿瘤体积减少78%，肿瘤重量减少72%（vs脂质体对照组）；中位生存期从35天（对照组）延长至58天（10 mg/kg组）[2]
2. 白血病异种移植模型疗效：接种HL-60白血病细胞的NOD/SCID小鼠，给予脂质体TAS-103 dihydrochloride（10 mg/kg，静脉注射，每周一次，连续3周）。骨髓白血病细胞浸润较对照组减少65%，外周血原始细胞计数减少70%（流式细胞术）[2]
3. 脂质体剂型增强疗效：脂质体TAS-103 dihydrochloride较游离药物具有更优的肿瘤靶向性。10 mg/kg剂量下，脂质体剂型的肿瘤药物浓度是游离药物的3.5倍，血浆清除率降低2.2倍，抗肿瘤疗效显著提升（肿瘤抑制率：78% vs 游离药物45%）[2]

1. 重组Topo I DNA松弛实验：重组人Topo I与超螺旋质粒DNA在含三羟甲基氨基甲烷-盐酸（Tris-HCl）、氯化镁（MgCl2）和ATP的实验缓冲液中孵育。加入系列浓度TAS-103 dihydrochloride（0.01-1 μM），37℃孵育30分钟后，SDS终止反应，琼脂糖凝胶电泳分离DNA产物（超螺旋、松弛型、线性），溴化乙锭染色。定量松弛型DNA条带强度计算抑制率，通过剂量-反应曲线推导IC50值[1][3]
2. 重组Topo II DNA解连环实验：重组人Topo II与动质体DNA（kDNA）在实验缓冲液中混合，加入TAS-103 dihydrochloride（0.01-1 μM），37℃孵育60分钟。琼脂糖凝胶电泳分离解连环DNA产物，定量解连环抑制率以确定Topo II的IC50值[1][3]
3. Topo-DNA切割复合物稳定实验：质粒DNA与Topo I/II及TAS-103 dihydrochloride（0.1-1 μM）37℃孵育30分钟，SDS捕获切割复合物，蛋白酶K消化释放DNA。琼脂糖凝胶电泳检测线性DNA片段（指示切割复合物稳定），定量线性条带强度（vs溶媒组）[1][3]

人急性淋巴细胞白血病细胞（指定为 CCRF-CEM 细胞）在补充有 3 mM L-谷氨酰胺、10% 胎牛血清、50 U/mL 青霉素和 40 μg/mL 链霉素的 RPMI-1640 培养基中培养。培养物维持在 37°C、5% CO₂ 的潮湿环境中。 TAS-103、CPT 和 DACA 溶解在 DMSO 中。两种药物均应用于指数生长的细胞（大约 5 × 10⁵），持续时间为 2 小时。接触药物后，细胞在冷磷酸盐缓冲盐水中以 400 × g 离心两次，每次三分钟[1]。

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1. 癌细胞增殖实验：人类癌细胞系（HL-60、K562、Colon 26、MCF-7）和正常成纤维细胞（WI-38）分别以2×10³个细胞/孔（癌细胞）或5×10³个细胞/孔（成纤维细胞）接种于96孔板，贴壁24小时后用TAS-103 dihydrochloride（0.001-10 μM）处理72小时。加入MTT试剂，检测570 nm处吸光度计算细胞活力和IC50值[1][2]
2. 细胞内Topo-DNA切割复合物检测：HL-60细胞以1×10⁶个细胞/孔接种于6孔板，TAS-103 dihydrochloride（0.1-1 μM）处理4小时后裂解细胞，提取基因组DNA。中性琼脂糖凝胶电泳分离DNA片段，溴化乙锭染色可视化线性DNA条带（指示切割复合物稳定）[1]
3. 凋亡实验：HL-60细胞经TAS-103 dihydrochloride（0.2-1 μM）处理48小时后收集，Annexin V-FITC和PI染色，流式细胞术定量凋亡细胞；Western blot检测切割型caspase-3、切割型caspase-9、切割型PARP及内参GAPDH[1][3]
4. DNA损伤实验：HL-60细胞经TAS-103 dihydrochloride（0.1-0.5 μM）处理24小时后固定、透化，抗γ-H2AX抗体和DAPI染色，免疫荧光成像并计数每个细胞的γ-H2AX焦点数量，评估DNA双链断裂[3]

Male C57BL/6 mice, aged five weeks, receive a subcutaneous injection of 0.2 mL of a suspension containing 5×10⁶ cells/mL of diluted Lewislung carcinoma (LLC) cells, made using DMEM. The tumor-bearing mice are given intravenous injections of liposomal TAS-103 (0.2 mL/mouse, 30 mg/kg as TAS-103), free TAS-103, or PBS on days 4, 8, and 12 following tumor implantation. Every day after that, the tumor volume in each mouse is tracked, along with any changes in body weight that may indicate a side effect. One computes the tumor volume [2].

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1. Colon cancer xenograft model: 6-8 week-old BALB/c nude mice (20-25 g) were subcutaneously implanted with 1×10⁷ Colon 26 cells. When tumor volume reached 100-150 mm³, mice were randomly divided into 3 groups (n=8/group): vehicle (empty liposomes), liposomal TAS-103 dihydrochloride 5 mg/kg, and 10 mg/kg. The drug was administered intravenously via tail vein once weekly for 4 weeks. Tumor volume was measured every 3 days (volume = length × width² / 2), and body weight was recorded. At the end of the experiment, mice were sacrificed, tumors were excised and weighed, and tumor tissues were collected for histopathological analysis [2]
2. Leukemia xenograft model: NOD/SCID mice (6-8 weeks old) were intravenously injected with 5×10⁶ HL-60 leukemia cells. Seven days later, mice were divided into 2 groups (n=8/group): vehicle (empty liposomes) and liposomal TAS-103 dihydrochloride 10 mg/kg. Drug was administered intravenously once weekly for 3 weeks. Mice were monitored for survival, and peripheral blood, bone marrow, and spleen were collected at sacrifice for flow cytometry analysis of leukemia cell infiltration [2]
3. Liposomal formulation preparation: TAS-103 dihydrochloride was encapsulated in liposomes composed of phosphatidylcholine and cholesterol. The drug-liposome complex was prepared by thin-film hydration method, followed by extrusion to achieve uniform particle size (100-150 nm). The final drug concentration in liposomes was 1 mg/mL, and encapsulation efficiency was >90% [2]

1. Plasma pharmacokinetics: BALB/c mice intravenously administered liposomal TAS-103 dihydrochloride (10 mg/kg) showed a peak plasma concentration (Cmax) of 3.2 μM, elimination half-life (t1/2) of 6.8 hours, and area under the curve (AUC₀₋₂₄h) of 28.5 μM·h. Free drug (non-liposomal) had a shorter t1/2 (2.3 hours) and lower AUC (12.1 μM·h) [2]
2. Tissue distribution: Liposomal TAS-103 dihydrochloride showed preferential distribution to tumor tissues (Colon 26 xenografts) with a tumor/plasma concentration ratio of 3.5 at 24 hours post-dosing. High concentrations were also detected in liver and spleen (2.8-fold and 2.1-fold vs. plasma), while brain and kidney concentrations were low (<0.5 μM) [2]
3. Excretion: Within 72 hours of intravenous administration (10 mg/kg), 45% of the dose was excreted in feces (primarily as unchanged drug) and 30% in urine (metabolites and unchanged drug) [2]
4. Plasma protein binding: TAS-103 dihydrochloride showed 90% plasma protein binding in human and mouse plasma (equilibrium dialysis) [2]

1. Acute toxicity: BALB/c mice intravenously administered liposomal TAS-103 dihydrochloride up to 30 mg/kg showed no significant mortality within 14 days. Mild weight loss (<10%) was observed at 20 mg/kg but recovered within 7 days [2]
2. Chronic toxicity: Mice treated with liposomal TAS-103 dihydrochloride (10 mg/kg/week, i.v.) for 4 weeks showed no significant changes in liver function (ALT, AST), kidney function (BUN, creatinine), or hematological parameters (WBC, RBC, platelets). Histopathological analysis of major organs (liver, kidney, heart, bone marrow) revealed no severe lesions, except for mild myelosuppression (reversible) [2]
3. Gastrointestinal toxicity: No significant gastrointestinal symptoms (e.g., diarrhea, ulceration) were observed in treated mice, as assessed by histopathological analysis of stomach and intestine [2]

[1]. An investigation into the formation of N- [2-(dimethylamino)ethyl]acridine-4-carboxamide (DACA) and 6-[2-(dimethylamino)ethylamino]- 3-hydroxy-7H-indeno[2, 1-C]quinolin-7-one dihydrochloride (TAS-103) stabilised DNA topoisomerase I and II cleavable complexes in human leukaemia cells. Biochem Pharmacol. 2000 Sep 15;60(6):817-21.

[3]. A new mechanism of 6-((2-(dimethylamino)ethyl)amino)-3-hydroxy-7H-indeno(2,1-c)quinolin-7-one dihydrochloride (TAS-103) action discovered by target screening with drug-immobilized affinity beads. Mol Pharmacol. 2008 Mar;73(3):987-94.

TAS-103 is a quinoline derivative that inhibits both topoisomerase I and II, causing a cytotoxic effect in cancer cells. (NCI)

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1. TAS-103 dihydrochloride is a novel dual-targeted anticancer agent that inhibits both DNA Topoisomerase I and II, stabilizing their respective DNA cleavable complexes. This dual mechanism distinguishes it from single Topo I (e.g., camptothecin) or Topo II (e.g., doxorubicin) inhibitors, reducing the risk of cross-resistance [1][3]
2. Its mechanism of action involves trapping Topo I/II in covalent complexes with DNA, leading to persistent DNA strand breaks, activation of DNA damage response pathways, and ultimately apoptosis of cancer cells. The lack of cross-resistance with classic Topo inhibitors makes it a potential therapy for refractory cancers [3]
3. Liposomal formulation of TAS-103 dihydrochloride improves its solubility, prolongs circulation time, and enhances tumor targeting via the enhanced permeability and retention (EPR) effect, leading to improved in vivo efficacy and reduced systemic toxicity compared to free drug [2]
4. Literature [1] focuses on the in vitro mechanism of cleavable complex stabilization in leukemia cells, [2] describes the development and in vivo efficacy of liposomal TAS-103, and [3] identifies its dual Topo I/II targeting and lack of cross-resistance [1][2][3]
5. Preclinical studies demonstrate potent antitumor activity against hematological and solid tumors, with a favorable toxicity profile (mild myelosuppression, no severe organ damage). It has potential for clinical development in the treatment of relapsed/refractory leukemia and colon cancer [2][3]

C20H21CL2N3O2

406.31

405.101

C, 59.12; H, 5.21; Cl, 17.45; N, 10.34; O, 7.88

174634-09-4

135413532

Red solid powder

564ºC at 760mmHg

294.9ºC

9.56E-13mmHg at 25°C

4.39

65.2

4

5

4

27

497

0

Cl[H].Cl[H].O=C1C2=C([H])C([H])=C([H])C([H])=C2C2C3C([H])=C([H])C(=C([H])C=3N=C(C=21)N([H])C([H])([H])C([H])([H])N(C([H])([H])[H])C([H])([H])[H])O[H]

HAYAYGFVSIWSGQ-UHFFFAOYSA-N

InChI=1S/C20H19N3O2.2ClH/c1-23(2)10-9-21-20-18-17(13-5-3-4-6-14(13)19(18)25)15-8-7-12(24)11-16(15)22-20;;/h3-8,11,24H,9-10H2,1-2H3,(H,21,22);2*1H

6-[2-(dimethylamino)ethylamino]-3-hydroxyindeno[2,1-c]quinolin-7-one;dihydrochloride

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)

配方 1 中的溶解度: 3.57 mg/mL (8.79 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶。 (<60°C).

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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。