Acyclic nucleoside phosphonates with adenine nucleobase inhibit Trypanosoma brucei adenine phosphoribosyltransferase in vitro.
Acyclic nucleoside phosphonates with adenine nucleobase inhibit Trypanosoma brucei adenine phosphoribosyltransferase in vitro.
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DOI：
10.1038/s41598-021-91747-6
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发表时间：
2021-06-25
影响因子：
4.6
通讯作者：
Zíková A
中科院分区：
文献类型：
--
作者：
Doleželová E;Klejch T;Špaček P;Slapničková M;Guddat L;Hocková D;Zíková A
所有医学上重要的单细胞原生动物都不能从头合成嘌呤，它们完全依赖嘌呤补救途径（PSP）生成核苷酸。因此，嘌呤衍生物被认为是抗寄生虫化合物的有希望的来源，因为它们可以作为PSP酶的抑制剂或在细胞内活化时作为毒性产物。在这里，我们的特点是布氏锥虫酶参与腺嘌呤，腺嘌呤磷酸核糖转移酶（APRT）的补救。我们发现，它的两种亚型（APRT 1和APRT 2）定位部分在胞质溶胶和部分在糖体的血流形式（BSF）的寄生虫。除非在以腺嘌呤作为唯一嘌呤来源的人工培养基中生长，否则两种APRT酶的RNA干扰沉默对BSF寄生虫的生长没有重大影响。为了添加到各种PSP酶的抑制剂组合中，我们设计了三种类型的无环核苷酸类似物作为潜在的APRT抑制剂。在15种抑制剂中，4种化合物抑制重组APRT 1的活性，Ki值为1 µM。ANP氨基磷酸酯膜可渗透的前药在基于细胞的测定中显示出显著的抗锥虫活性，尽管事实上APRT酶对于T.布氏杆菌体外生长虽然这表明所测试的ANP前药通过其他方式在T.布鲁氏菌，新设计的抑制剂可以进一步改进和探索以鉴定它们的实际靶点。
All medically important unicellular protozoans cannot synthesize purines de novo and they entirely rely on the purine salvage pathway (PSP) for their nucleotide generation. Therefore, purine derivatives have been considered as a promising source of anti-parasitic compounds since they can act as inhibitors of the PSP enzymes or as toxic products upon their activation inside of the cell. Here, we characterized a Trypanosoma brucei enzyme involved in the salvage of adenine, the adenine phosphoribosyl transferase (APRT). We showed that its two isoforms (APRT1 and APRT2) localize partly in the cytosol and partly in the glycosomes of the bloodstream form (BSF) of the parasite. RNAi silencing of both APRT enzymes showed no major effect on the growth of BSF parasites unless grown in artificial medium with adenine as sole purine source. To add into the portfolio of inhibitors for various PSP enzymes, we designed three types of acyclic nucleotide analogs as potential APRT inhibitors. Out of fifteen inhibitors, four compounds inhibited the activity of the recombinant APRT1 with Ki in single µM values. The ANP phosphoramidate membrane-permeable prodrugs showed pronounced anti-trypanosomal activity in a cell-based assay, despite the fact that APRT enzymes are dispensable for T. brucei growth in vitro. While this suggests that the tested ANP prodrugs exert their toxicity by other means in T. brucei, the newly designed inhibitors can be further improved and explored to identify their actual target(s).
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影响因子：
9.6
作者：
Boitz JM;Ullman B;Jardim A;Carter NS
通讯作者：
Carter NS
影响因子：
6.7
作者：
Klejch, Tomas;Keough, Dianne T.;Hockova, Dana
通讯作者：
Hockova, Dana
影响因子：
4.6
作者：
Benz C;Dondelinger F;McKean PG;Urbaniak MD
通讯作者：
Urbaniak MD
影响因子：
3.8
作者：
Dolezelova, Eva;Teran, David;Zikova, Alena
通讯作者：
Zikova, Alena
影响因子：
2.9
作者：
Bashor, C;Denu, JM;Ullman, B
通讯作者：
Ullman, B