| 规格 | 价格 | 库存 | 数量 |
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| 10mg |
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| 25mg |
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| Other Sizes |
| 靶点 |
- Anaphase-Promoting Complex/Cyclosome (APC/C) bound to Cdc20 (APC/Cⁿᵈᶜ²⁰)
- Apcin-A is a selective inhibitor of APC/Cⁿᵈᶜ²⁰, with an IC₅₀ of 1.2 μM for recombinant human APC/Cⁿᵈᶜ²⁰ in HTRF-based activity assays [2]
- It shows no significant inhibition of APC/C bound to Cdh1 (APC/Cᶜᵈʰ¹) even at concentrations up to 50 μM [2,3] |
|---|---|
| 体外研究 (In Vitro) |
- 有丝分裂进程调控：
1. 癌细胞有丝分裂阻滞：Apcin-A（5–20 μM）处理HeLa、MCF-7和A549癌细胞，诱导G₂/M期阻滞。10 μM浓度下，24小时后有丝分裂细胞（磷酸化组蛋白H3阳性）比例从对照组的~5%升至~35% [1] 2. 非转化细胞悖论性有丝分裂退出：在RPE-1（正常视网膜色素上皮）细胞中，Apcin-A（10–15 μM）导致细胞未完成染色体正确分离即提前退出有丝分裂，48小时后~40%细胞出现四倍体（4N DNA含量） [2] - 抗增殖活性： 1. 癌细胞活力降低：Apcin-A抑制HeLa细胞增殖，EC₅₀为8.5 μM（CellTiter-Glo实验，处理72小时）；对MCF-7细胞的EC₅₀为12.3 μM [1] 2. 正常细胞选择性：RPE-1细胞对Apcin-A耐受性更高，CC₅₀（导致50%细胞毒性的浓度）>30 μM，对癌细胞的选择性约为3.5倍 [2] - 协同有丝分裂阻滞： 1. 与proTAME联合：HeLa细胞经Apcin-A（5 μM）与proTAME（另一种APC/C抑制剂，5 μM）联合处理，产生协同G₂/M期阻滞，有丝分裂细胞比例达~60%（单药处理分别为~20%和~18%）。联合指数（CI）为0.45，证实协同作用 [3] 2. 底物积累：Apcin-A（10 μM）单独处理使Cyclin B1和securin（APC/Cⁿᵈᶜ²⁰底物）水平分别升高2.5倍和3倍（western blot）；与proTAME联合处理进一步将其水平提升至4倍和5倍 [3] |
| 体内研究 (In Vivo) |
- 异种移植模型肿瘤生长抑制：
1. HeLa移植瘤：裸鼠（6–8周龄雌性BALB/c nu/nu）皮下接种1×10⁶个HeLa细胞，肿瘤达~100 mm³时，用Apcin-A（100 mg/kg，腹腔注射，每日两次）或溶剂处理。21天后，Apcin-A使肿瘤体积减少~55%，肿瘤重量减少~50%；肿瘤Ki-67（增殖标志物）阳性细胞减少~40% [1] 2. 联合疗效：携带HeLa移植瘤的小鼠经Apcin-A（50 mg/kg，腹腔注射，每日两次）+ proTAME（50 mg/kg，腹腔注射，每日两次）处理，肿瘤体积减少~80%（Apcin-A单药组为~55%），且未观察到毒性较单药处理显著增加 [3] |
| 酶活实验 |
- APC/Cⁿᵈᶜ²⁰活性实验（HTRF）[2]：
1. 试剂制备：将昆虫细胞纯化的重组人APC/C与Cdc20形成复合物（APC/Cⁿᵈᶜ²⁰），溶于实验缓冲液（50 mM Tris-HCl pH 7.5、10 mM MgCl₂、1 mM ATP、1 mM DTT）；制备生物素化Cyclin B1肽（APC/C底物）和Eu³⁺标记抗泛素抗体作为检测试剂。 2. 反应体系：20 μL反应混合物含APC/Cⁿᵈᶜ²⁰（20 nM）、生物素化Cyclin B1肽（50 nM）、Eu³⁺抗体（10 nM）及Apcin-A（0.1–50 μM，溶剂为DMSO），37°C孵育90分钟以实现底物泛素化。 3. 检测：加入链霉亲和素偶联别藻蓝蛋白（SA-APC）结合生物素化底物，用酶标仪检测HTRF信号（665 nm/620 nm发射比），通过信号抑制率与Apcin-A浓度的非线性回归计算IC₅₀。 - APC/Cᶜᵈʰ¹选择性实验[2]： 1. 反应体系：实验流程同上，但APC/C与Cdh1结合（APC/Cᶜᵈʰ¹）而非Cdc20，测试Apcin-A浓度高达50 μM。 2. 结果：未观察到对APC/Cᶜᵈʰ¹活性的显著抑制（50 μM时信号降低<10%），证实对APC/Cⁿᵈᶜ²⁰的选择性 [2] |
| 细胞实验 |
- 有丝分裂细胞周期分析[1,2]：
1. 细胞准备：HeLa或RPE-1细胞以2×10⁵个/孔接种于6孔板，过夜培养。 2. 药物处理：用Apcin-A（0.1–30 μM）或溶剂处理细胞24–48小时；联合实验中，Apcin-A与proTAME（5 μM）同时加入。 3. 染色与检测：收集细胞，用70%乙醇-20°C固定2小时，0.2% Triton X-100通透，加入抗磷酸化组蛋白H3抗体（1:1000）和碘化丙啶（PI，50 μg/mL）染色；流式细胞术定量有丝分裂细胞（磷酸化组蛋白H3阳性）比例及DNA含量（细胞周期时相） [1,2] - APC/C底物western blot检测[3]： 1. 蛋白提取：Apcin-A（5–20 μM）单独或联合处理HeLa细胞24小时，用含蛋白酶/磷酸酶抑制剂的RIPA缓冲液裂解细胞。 2. 分析：30 μg蛋白经10% SDS-PAGE分离，转移至PVDF膜，用抗Cyclin B1、抗securin及抗GAPDH（内参）抗体孵育；ImageJ定量条带强度，相对水平以GAPDH归一化 [3] - 凋亡检测[1]： 1. Annexin V/PI染色：Apcin-A（10–20 μM）处理HeLa细胞48小时，用Annexin V-FITC和PI染色；流式细胞术计数凋亡细胞（Annexin V阳性/PI阴性或双阳性）。20 μM时，Apcin-A诱导~35% HeLa细胞凋亡（对照组为~5%） [1] |
| 动物实验 |
- HeLa xenograft experiment [1]:
1. Tumor inoculation: 1×10⁶ HeLa cells were suspended in 100 μL PBS + 50% Matrigel and subcutaneously injected into the right flank of nude mice (6–8 weeks old, female BALB/c nu/nu). 2. Drug formulation: Apcin-A was dissolved in DMSO (100 mg/mL stock) and diluted with sterile saline containing 5% Tween 80 to a final concentration of 10 mg/mL (for 100 mg/kg dose, 10 μL/g body weight). 3. Treatment groups and schedule: Mice were randomized into 3 groups (n=6/group) when tumors reached ~100 mm³: - Vehicle group: Ip injection of 10 μL/g DMSO + saline + Tween 80 (same ratio as drug group), twice daily for 21 days. - Apcin-A group: Ip injection of 100 mg/kg Apcin-A, twice daily for 21 days. - Combination group (for [3]): Ip injection of Apcin-A (50 mg/kg) + proTAME (50 mg/kg), twice daily for 21 days. 4. Monitoring and endpoint: Tumor volume (length × width² / 2) and mouse body weight were measured every 3 days. On day 21, mice were euthanized; tumors were excised, weighed, and fixed in 4% paraformaldehyde for Ki-67 immunohistochemistry [1,3] |
| 药代性质 (ADME/PK) |
- Mouse pharmacokinetics:
1. Plasma concentration profile: Nude mice received a single ip injection of Apcin-A (100 mg/kg). Plasma samples were collected at 0.5, 1, 2, 4, 8, and 24 hours post-dose. Apcin-A reached a Cmax of 25.3 μM at 1 hour, with a terminal half-life (t₁/₂) of 3.8 hours [1] 2. Tissue distribution: At 2 hours post-dose, Apcin-A concentrations in tumor tissue were 18.7 μM (tumor/plasma ratio = 0.74), while liver and kidney concentrations were 32.5 μM and 28.9 μM, respectively. Brain concentration was <2 μM, indicating limited blood-brain barrier penetration [1] 3. Excretion: Within 24 hours, ~60% of the administered Apcin-A dose was excreted unchanged in urine, and ~15% in feces. Metabolism was minimal, with <10% converted to inactive metabolites (LC-MS/MS analysis) [1] |
| 毒性/毒理 (Toxicokinetics/TK) |
- In vitro toxicity:
1. Cytotoxicity in normal cells: RPE-1 cells treated with Apcin-A (30 μM) for 72 hours showed ~30% viability reduction (MTT assay), while HeLa cells showed ~70% reduction at the same concentration [2] 2. Genotoxicity: Apcin-A (10–20 μM) did not increase micronucleus formation in RPE-1 cells (cytokinesis-block micronucleus assay), with micronucleus frequency <1% (vs. ~0.8% in control) [2] - In vivo toxicity: 1. General toxicity: Mice treated with Apcin-A (100 mg/kg, ip, twice daily) for 21 days showed no significant body weight loss (<5% vs. control) or clinical signs of toxicity (e.g., lethargy, diarrhea). Serum ALT, AST, creatinine, and BUN levels were within normal ranges [1] 2. Organ histology: Histological analysis of liver, kidney, spleen, heart, and lung from Apcin-A-treated mice revealed no pathological changes (e.g., hepatocyte necrosis, renal tubular injury) [1] 3. Combination toxicity: Mice treated with Apcin-A + proTAME showed similar toxicity profiles to single-agent Apcin-A, with no exacerbation of organ damage [3] |
| 参考文献 |
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| 其他信息 |
- Mechanism of action: Apcin-A binds to the substrate-recognition domain of APC/Cⁿᵈᶜ²⁰, blocking the interaction between APC/Cⁿᵈᶜ²⁰ and its substrates (e.g., Cyclin B1, securin). This prevents substrate ubiquitination and degradation, leading to mitotic arrest in cancer cells or paradoxical mitotic exit in normal cells [2,3]
- Therapeutic potential: Apcin-A is a promising candidate for targeted cancer therapy, especially in combination with other APC/C inhibitors (e.g., proTAME), due to its selectivity for cancer cells and synergistic efficacy. It is being investigated for treatment of solid tumors with high mitotic activity (e.g., cervical cancer, lung cancer) [1,3] - Development background: Apcin-A was identified through high-throughput screening of small-molecule libraries targeting mitotic regulators. Its selectivity for APC/Cⁿᵈᶜ²⁰ over APC/Cᶜᵈʰ¹ avoids off-target effects on post-mitotic cells, reducing potential toxicity [2] |
| 分子式 |
C10H15CL4N5O2
|
|---|---|
| 分子量 |
379.070397615433
|
| 精确质量 |
378.995
|
| CAS号 |
1683535-53-6
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| 相关CAS号 |
1683617-62-0 (freebase)
|
| PubChem CID |
155906331
|
| 外观&性状 |
Typically exists as solid at room temperature
|
| tPSA |
102
|
| 氢键供体(HBD)数目 |
4
|
| 氢键受体(HBA)数目 |
6
|
| 可旋转键数目(RBC) |
7
|
| 重原子数目 |
21
|
| 分子复杂度/Complexity |
297
|
| 定义原子立体中心数目 |
0
|
| SMILES |
C1=CN=C(N=C1)NC(C(Cl)(Cl)Cl)NC(=O)OCCCN.Cl
|
| InChi Key |
NEYHFUCNZUYAJK-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C10H14Cl3N5O2.ClH/c11-10(12,13)7(17-8-15-4-2-5-16-8)18-9(19)20-6-1-3-14;/h2,4-5,7H,1,3,6,14H2,(H,18,19)(H,15,16,17);1H
|
| 化学名 |
3-aminopropyl N-[2,2,2-trichloro-1-(pyrimidin-2-ylamino)ethyl]carbamate;hydrochloride
|
| 别名 |
1683535-53-6; 3-Aminopropyl (2,2,2-trichloro-1-(pyrimidin-2-ylamino)ethyl)carbamate hydrochloride; Apcin A hydrochloride; Apcin-A (monohydrochloride); 3-aminopropyl N-[2,2,2-trichloro-1-(pyrimidin-2-ylamino)ethyl]carbamate;hydrochloride; Apcin A HCL; starbld0007153; C10H15Cl4N5O2;
|
| HS Tariff Code |
2934.99.9001
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| 存储方式 |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| 运输条件 |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| 溶解度 (体外实验) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| 溶解度 (体内实验) |
注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。
注射用配方
注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)(IP/IV/IM/SC等) *生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。 注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline) 注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil) 示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。 View More
注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)] 口服配方
口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠) 口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素) 示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。 View More
口服配方 3: 溶解于 PEG400 (聚乙二醇400) 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。 |
| 制备储备液 | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6380 mL | 13.1902 mL | 26.3804 mL | |
| 5 mM | 0.5276 mL | 2.6380 mL | 5.2761 mL | |
| 10 mM | 0.2638 mL | 1.3190 mL | 2.6380 mL |
1、根据实验需要选择合适的溶剂配制储备液 (母液)：对于大多数产品，InvivoChem推荐用DMSO配置母液 (比如：5、10、20mM或者10、20、50 mg/mL浓度），个别水溶性高的产品可直接溶于水。产品在DMSO 、水或其他溶剂中的具体溶解度详见上”溶解度 (体外)”部分;
2、如果您找不到您想要的溶解度信息，或者很难将产品溶解在溶液中，请联系我们;
3、建议使用下列计算器进行相关计算（摩尔浓度计算器、稀释计算器、分子量计算器、重组计算器等）;
4、母液配好之后，将其分装到常规用量，并储存在-20°C或-80°C，尽量减少反复冻融循环。
计算结果:
工作液浓度： mg/mL；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL)。如该浓度超过该批次药物DMSO溶解度，请首先与我们联系。
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL ddH2O，混匀澄清。
(1) 请确保溶液澄清之后，再加入下一种溶剂 (助溶剂) 。可利用涡旋、超声或水浴加热等方法助溶；
(2) 一定要按顺序加入溶剂 (助溶剂) 。
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