(+)-Aphidicolin
(Synonyms: 阿非迪霉素) 目录号 : GC10867Aphidicolin ((+)-Aphidicolin), a reversible inhibitor of eukaryotic nuclear DNA replication, can block the cell cycle at the pre-S phase[1].
Cas No.:38966-21-1
Sample solution is provided at 25 µL, 10mM.
;)
,-1-7$$$37316672.jpg');)
;)
;)
$$$36806353.jpg');)
;33(7)%EF%BC%9A546-561$$$37156877.jpg');)
;)
;)
;)
%201124-1138.$$$35908550.jpg');)
%20766-780.$$$37100057.jpg');)
%20418-432.$$$36893736.jpg');)
%EF%BC%9A-240-255.$$$35108512.jpg');)
533-545$$$36543525.jpg');)
%201-13.$$$36600053.jpg');)
;)
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
|
Cell lines |
HeLa cells |
|
Preparation Method |
HeLa cells were synchronized by 24 h incubation with aphidicolin. After aphidicolin removal, cells were collected at different time stamps (0, 1.5, 4, 8, 12 and 24 h). DNA content was measured by flow cytometry by tracing the fluorescence intensity of cells. |
|
Reaction Conditions |
1 mg/ml; 0, 1.5, 4, 8, 12 and 24 h |
|
Applications |
Aphidicolin efficiently synchronizes HeLa cells. Late S/G2 phase was reached around 12 hours after aphidicolin removal. After 24 hours cells return to their original, unsynchronized distribution. |
| Animal experiment [2]: | |
|
Animal models |
2-week-old WT and DKO (E2F1/E2F2 double-knockout) mice |
|
Preparation Method |
In vivo BrdU labeling and γ-H2AX accumulation in pancreases derived from 2-week-old WT and DKO mice, after i.p. injection of vehicle (saline) or aphidicolin. |
|
Dosage form |
10 µg/g; i.p. |
|
Applications |
Treatment with aphidicolin (10 µg/g) completely abolishes the incorporation of BrdU and reduces the number of cells with pan-nuclear γ-H2AX staining in DKO cells. |
|
References: Szczepański K, et al. Stability of cytoplasmic nanoviscosity during cell cycle of HeLa cells synchronized with Aphidicolin. Sci Rep. 2019 Nov 11;9(1):16486. | |
Aphidicolin ((+)-Aphidicolin), a reversible inhibitor of eukaryotic nuclear DNA replication, can block the cell cycle at the pre-S phase[1]. The minimal concentration of aphidicolin required to produce reversible inhibition of DNA replication without toxic effects in embryos to be 0.5 µM[2].
In vitro experiment it shown that treatment with APC (Aphidicolin) at 6 μM in PBMC cell did not impact baseline DNA damage but could reliably block DNA repair after H2O2 challenge in both fresh and cryopreserved samples[3]. In vitro, treatment with 20 μM aphidicolin for 24 h in bovine aortic ECs (BAECs) reduced BAEC viability by ~40%, which was accompanied by increased NO production, phosphorylation of eNOS at Ser1179 (p-eNOS-Ser1179 were not altered by 1,2-bis (2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl ester) (BAPTA-AM), a cell permeable and specific intracellular Ca2+ chelator[4]. In addition, 5 x 10(-7) M aphidicolin can kill all cells of four human neuroblastoma cell lines as an agent which selectively kills neuroblastoma cells in vitro [5].
In vivo efficacy test it indicated that 4.5 mg aphidicolin/kg body weight was the maximum aphidicolin dose that could be applied as liposomal preparation that tested in neuroblastoma-bearing (UKF-NB-3) mice[6]. Aphidicolin glycinate (has higher solubility agent) (100 mg/kg; i.p.; once every 3 hfor 9 days) showed anti-tumor activity against the implanted B16 melanoma and M5076 sarcoma in murine, producing maximum increased life spans of 75% [7].
阿非迪霉素是真核DNA复制的可逆抑制剂，可阻断前S期的细胞周期[1]。在胚胎中产生对DNA复制的可逆抑制而不产生毒性作用所需的阿非迪霉素最低浓度为0.5 µM[2]。
体外实验表明，在新鲜和冷冻保存的样品中，6 μM的阿非迪霉素在PBMC细胞中处理不会影响基线DNA损伤，但可以可靠地阻断H2O2激发后的DNA修复[3]。在体外，在牛主动脉内皮细胞中用20 μM阿非迪霉素处理24小时，使其活力降低了约40%，同时NO产生增加，Ser1179（p-eNOS-Ser1179）处的eNOS磷酸化没有被1,2-二（2-氨基苯氧基）乙烷-N，N，N'，N'-四乙酸四（乙酰氧基甲酯）（BAPTA-AM）（一种细胞可渗透的特异性细胞内Ca2+螯合剂）改变[4]。此外，作为一种在体外选择性杀死神经母细胞癌细胞的药剂，5 x 10（-7）M 阿非迪霉素可以杀死四种人类神经母细胞瘤细胞系的所有细胞[5]。
体内疗效测试表明，4.5 mg 阿非迪霉素/kg体重是可作为脂质体制剂应用于神经母细胞瘤（UKF-NB-3）小鼠的最大剂量[6]。阿非迪霉素（缩水甘油，具有更高的溶解剂）（100 mg/kg；腹腔注射；每3 h或9天一次）对小鼠植入的B16黑色素瘤和M5076肉瘤显示出抗肿瘤活性，最大寿命延长75% [7]。
References:
[1] Zhang T.Y., et al. Positive effects of treatment of donor cells with aphidicolin on the preimplantation development of somatic cell nuclear transfer embryos in Chinese Bama mini-pig (Sus Scrofa) Anim. Sci. J. 2012;83:103–110.
[2] Navarro-Serna S, et al. Effect of Aphidicolin, a Reversible Inhibitor of Eukaryotic Nuclear DNA Replication, on the Production of Genetically Modified Porcine Embryos by CRISPR/Cas9. Int J Mol Sci. 2022 Feb 15;23(4):2135.
[3] Odongo GA, et al. Optimization of the alkaline comet assay for easy repair capacity quantification of oxidative DNA damage in PBMC from human volunteers using aphidicolin block. DNA Repair (Amst). 2019 May;77:58-64.
[4] Park JH, et al. Nuclear localization of endothelial nitric oxide synthase and nitric oxide production attenuates aphidicolin-induced endothelial cell death. Nitric Oxide. 2021 May 1;109-110:12-19.
[5] Cinatl J, et al. Aphidicolin selectively kills neuroblastoma cells in vitro. Cancer Lett. 1992 Dec 24;67(2-3):199-206.
[6] Michaelis M, et al. Increased systemic efficacy of aphidicolin encapsulated in liposomes. Oncol Rep. 2005 Jan;13(1):157-60.
[7] O'Dwyer PJ, et al. Antitumor activity and biochemical effects of aphidicolin glycinate (NSC 303812) alone and in combination with cisplatin in vivo. Cancer Res. 1994 Feb 1;54(3):724-9.
| Cas No. | 38966-21-1 | SDF | |
| 别名 | 阿非迪霉素 | ||
| 化学名 | (3R,4R,4aR,6aS,8R,9R,11aS,11bS)-4,9-bis(hydroxymethyl)-4,11b-dimethyltetradecahydro-8,11a-methanocyclohepta[a]naphthalene-3,9-diol | ||
| Canonical SMILES | O[C@@]1(CC[C@@]23[C@@]([H])(CC[C@]4([C@@](C)([C@@H](CC[C@@]43C)O)CO)[H])C[C@@H]1C2)CO | ||
| 分子式 | C20H34O4 | 分子量 | 338.48 |
| 溶解度 | 50mg/mL in DMSO, 10mg/mL in methanol | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
||
| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 2.9544 mL | 14.7719 mL | 29.5438 mL |
| 5 mM | 0.5909 mL | 2.9544 mL | 5.9088 mL |
| 10 mM | 0.2954 mL | 1.4772 mL | 2.9544 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
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，
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 saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。