DNA sequence-selective adenine alkylation, mechanism of adduct repair, and in vivo antitumor activity of the novel achiral seco-amino-cyclopropylbenz[e]indolone analogue of duocarmycin AS-I-145
DNA sequence-selective adenine alkylation, mechanism of adduct repair, and in vivo antitumor activity of the novel achiral seco-amino-cyclopropylbenz[e]indolone analogue of duocarmycin AS-I-145
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DOI：
10.1158/1535-7163.mct-07-0294
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发表时间：
2007-10-01
影响因子：
5.7
通讯作者：
Hartley, John A.
中科院分区：
文献类型：
--
作者：
Kiakos, Konstantinos;Sato, Atsushi;Hartley, John A.
AS-I-145是一种新的倍癌霉素的非手性开环-氨基-环丙基苯-z [e]吲哚酮（开环-氨基-CBI）类似物，其是从设计CC-1065/倍癌霉素药物的替代策略发展而来的，缺乏天然药物的特征手性中心。通过Taq聚合酶终止试验评估该化合物的序列特异性，鉴定质粒DNA上的共价修饰位点。腺嘌呤-N3加合物在AT-丰富的序列，使用热诱导链切割试验确认。这些研究表明，该化合物保留了相关天然化合物的固有序列选择性。评估了缺失切除和复制后修复（PRR）途径的酵母突变体的ASI-145敏感性。敏感性曲线表明，序列特异性腺嘌呤-N3加合物是核苷酸切除修复（NER）的底物，而不是碱基切除修复（BER）。采用单链连接PCR跟踪酵母细胞中核苷酸解析时损伤的诱导和修复。序列特异性保存在完整的细胞，和加合物消除发生在转录偶联的方式，并依赖于一个功能性NER途径和Rad 18。在哺乳动物DNA修复突变细胞中证实了NER作为主要切除途径的参与。AS-I-145在国家癌症研究所标准中空纤维测定中显示出良好的体内抗肿瘤活性，并且当静脉内或口服给药时，对人乳腺MDA-MD-435异种移植物具有活性。其新颖的结构和体内活性使得AS-I-145成为设计CC-1065和倍癌霉素的新型非手性类似物的新范例。
AS-I-145 is a novel achiral seco-amino-cyclopropylben-z[e]indolone (seco-amino-CBI) analogue of duocarmycin that has evolved from an alternative strategy of designing CC-1065/duocarmycin agents lacking the characteristic chiral center of the natural agents. The sequence specificity of this compound was assessed by a Taq polymerase stop assay, identifying the sites of covalent modification on plasmid DNA. The adenine-N3 adducts were confirmed at AT-rich sequences using a thermally induced strand cleavage assay. These studies reveal that this compound retains the inherent sequence selectivity of the related natural compounds. The ASI-145 sensitivity of yeast mutants deficient in excision and post-replication repair (PRR) pathways was assessed. The sensitivity profile suggests that the sequence-specific adenine-N3 adducts are substrates for nucleotide excision repair (NER) but not base excision repair (BER). Single-strand ligation PCR was employed to follow the induction and repair of the lesions at nucleotide resolution in yeast cells. Sequence specificity was preserved in intact cells, and adduct elimination occurred in a transcription-coupled manner and was dependent on a functional NER pathway and Rad18. The involvement of NER as the predominant excision pathway was confirmed in mammalian DNA repair mutant cells. AS-I-145 showed good in vivo antitumor activity in the National Cancer Institute standard hollow fiber assay and was active against the human breast MDA-MD-435 xenograft when administered i.v. or p.o. Its novel structure and in vivo activity renders AS-I-145 a new paradigm in the design of novel achiral analogues of CC-1065 and the duocarmycins.