(+)-Aphidicolin

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].

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.

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]。