Deeken John F, Robey Robert W, Shukla Suneet, Steadman Kenneth, Chakraborty Arup R, Poonkuzhali Balasubramanian, Schuetz Erin G, Holbeck Susan, Ambudkar Suresh V, Bates Susan E
Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Washington, DC 20007, USA.
Mol Pharmacol. 2009 Nov;76(5):946-56. doi: 10.1124/mol.109.056192. Epub 2009 Jul 24.
ABCG2 is an ATP-binding cassette transporter that counts multiple anticancer compounds among its substrates and is believed to regulate oral bioavailability as well as serve a protective role in the blood-brain barrier, the maternal-fetal barrier, and hematopoietic stem cells. We sought to determine whether novel compounds that interact with the transporter could be identified through analysis of cytotoxicity profiles recorded in the NCI Anticancer Drug Screen database. A flow cytometric assay was used to measure ABCG2 function in the 60 cell lines and generate a molecular profile for COMPARE analysis. This strategy identified >70 compounds with Pearson correlation coefficients (PCCs) >0.4, where reduced drug sensitivity correlated with ABCG2 expression, as well as >120 compounds with PCCs < -0.4, indicating compounds to which ABCG2 expression conferred greater sensitivity. Despite identification of known single nucleotide polymorphisms in the ABCG2 gene in a number of the cell lines, omission of these lines from the COMPARE analysis did not affect PCCs. Available compounds were subjected to validation studies to confirm interaction with the transporter, including flow cytometry, [(125)I]IAAP binding, and cytotoxicity assays, and interaction was documented in 20 of the 27 compounds studied. Although known substrates of ABCG2 such as mitoxantrone or topotecan were not identified, we characterized three novel substrates-5-hydroxypicolinaldehyde thiosemicarbazone (NSC107392), (E)-N-(1-decylsulfanyl-3-hydroxypropan-2-yl)-3-(6-methyl-2,4-dioxo-1H-pyrimidin-5-yl)prop-2-enamide (NSC265473), and 1,2,3,4,7-pentahydroxy-1,3,4,4a,5,11b-hexahydro[1,3]dioxolo[4,5-j]phenanthridin-6(2H)-one [NSC349156 (pancratistatin)]-and four compounds that inhibited transporter function-2-[methyl(2-pyridin-2-ylethyl)-amino]fluoren-9-one hydroiodide (NSC24048), 5-amino-6-(7-amino-5,8-dihydro-6-methoxy-5,8-dioxo-2-quinolinyl)-4-(2-hydroxy-3,4-dimethoxyphenyl)-3-methyl-2-pyridinecarboxylic acid, methyl ester (NSC45384), (17beta)-2,4-dibromo-estra-1,3,5(10)-triene-3,17-diol (NSC103054), and methyl N-(pyridine-4-carbonylamino)carbamodithioate (NSC636795). In summary, COMPARE analysis of the NCI drug screen database using the ABCG2 functional profile was able to identify novel substrates and transporter-interacting compounds.
ABCG2是一种ATP结合盒转运蛋白，其底物包括多种抗癌化合物，被认为可调节口服生物利用度，并在血脑屏障、母胎屏障和造血干细胞中发挥保护作用。我们试图通过分析美国国立癌症研究所（NCI）抗癌药物筛选数据库中记录的细胞毒性谱，来确定是否能鉴定出与该转运蛋白相互作用的新型化合物。采用流式细胞术检测60种细胞系中的ABCG2功能，并生成用于COMPARE分析的分子谱。该策略鉴定出70多种皮尔逊相关系数（PCC）>0.4的化合物，其中药物敏感性降低与ABCG2表达相关，还有120多种PCC<-0.4的化合物，表明ABCG2表达使这些化合物具有更高的敏感性。尽管在许多细胞系中鉴定出了ABCG2基因中已知的单核苷酸多态性，但在COMPARE分析中省略这些细胞系并不影响PCC。对现有化合物进行验证研究，以确认其与转运蛋白的相互作用，包括流式细胞术、[（125）I]IAAP结合和细胞毒性测定，在所研究的27种化合物中有20种记录到了相互作用。虽然未鉴定出ABCG2的已知底物如米托蒽醌或拓扑替康，但我们鉴定出了三种新型底物——5-羟基吡啶甲醛硫代半卡巴腙（NSC107392）、（E）-N-（1-癸基硫烷基-3-羟基丙-2-基）-3-（6-甲基-2,4-二氧代-1H-嘧啶-5-基）丙-2-烯酰胺（NSC265473）和1,2,3,4,7-五羟基-1,3,4,4a,5,11b-六氢[1,3]二氧杂环戊烯并[4,5-j]菲啶-6（2H）-酮[NSC349156（ Pancratistatin）]——以及四种抑制转运蛋白功能的化合物——2-[甲基（2-吡啶-2-基乙基）-氨基]芴-9-酮氢碘酸盐（NSC24048）、5-氨基-6-（7-氨基-5,8-二氢-6-甲氧基-5,8-二氧代-2-喹啉基）-4-（2-羟基-3,4-二甲氧基苯基）-3-甲基-2-吡啶羧酸甲酯（NSC45384）、（17β）-2,4-二溴-雌甾-1,3,5（10）-三烯-3,17-二醇（NSC103054）和N-（吡啶-4-羰基氨基）氨基二硫代甲酸甲酯（NSC636795）。总之，使用ABCG2功能谱对NCI药物筛选数据库进行COMPARE分析，能够鉴定出新型底物和与转运蛋白相互作用的化合物。
