Small molecule inhibitors pantothenate synthesis against M. tuberculosis

描述（由申请人提供）：本R21探索性提案在合理开发可能有效缩短结核病治疗持续时间和/或改善耐多药结核病治疗的药物中使用了综合靶向方法。在结核病研究界存在广泛的共识，即缩短药物治疗持续时间而不增加复发率的关键在于特异性靶向对结核分枝杆菌的非复制持久性（NRP）至关重要的蛋白质。一种这样的靶标是泛酸合成酶（PS）。研究表明，PS对于小鼠的持续生长至关重要，并且在NRP的体外模型中过表达。最近成功的PS蛋白的结构阐明和PS的功能测定的可用性开辟了一条道路，应用计算机辅助药物设计方法与药物化学的努力，以发现新的治疗剂NRP TB。我们假设，这将是可能的，以确定和设计抑制剂的PS使用相结合的计算机辅助药物设计和药物化学。将在全细胞水平上筛选在低微摩尔范围内显示出抗PS活性的那些化合物的杀死复制和NRP M的能力。在基于韦恩低氧模型的微孔板测定中，以及在哺乳动物细胞毒性方面的选择性，测定了结核病。这种反复的过程有望产生几种低微摩尔的、铅或药物样的PS抑制剂。这些线索将是进一步开发的候选者，包括更广泛的药物化学工作和后续项目中NRP动物模型的评价。为实现这一目标，我们的具体目标如下：（1）通过寻找PS天然底物的类似物、对接商业化合物数据库、从头/合理药物设计、虚拟聚焦组合库等方法，筛选出PS的先导或药物样低分子量抑制剂;合成最佳的先导化合物和先导化合物的类似物（如果它们不能商购获得）;（2）测定候选药物对PS功能活性的抑制。(3)鉴定对活跃生长和/或非复制型持久性（NRP）结核分枝杆菌显示出显著选择活性的化合物。根据活性、细胞毒性和产生的选择性指数进行优先排序。

DESCRIPTION (provided by applicant): This R21 exploratory proposal uses an integrated target-oriented approach in the rational development of drugs that may effectively shorten the duration of therapy in tuberculosis and/or improve the treatment of multi-drug resistant TB. There exists a broad consensus in the TB research community that the key to shortening the duration of drug therapy without an increase in relapse rates lies in the specific targeting of proteins that are essential to the non-replicating persistence (NRP) of Mycobacterium tuberculosis. One such target is Pantothenate synthetase (PS). It has been shown that PS is crucial for persistent growth in mice and is over-expressed in in vitro models of NRP. Recent success in the structure elucidation of the PS protein and the availability of functional assays for PS opens a way to apply computer-aided drug design methods in concert with medicinal chemistry efforts to discover new therapeutic agents for NRP TB. We hypothesize that it would be possible to identify and design inhibitors for PS using a combination of computer-aided drug design and medicinal chemistry. Those compounds demonstrating activity at low micromolar range against PS will be screened at the whole cell level for the ability to kill replicating and NRP M. tuberculosis in a microplate assay based on the Wayne low oxygen model and for selectivity with respect to mammalian cell toxicity. This iterative process is expected to yield several low micromolar, lead- or drug- like inhibitors of PS. Such leads would be candidates for further development including more extensive medicinal chemistry efforts and the evaluation in animal models of NRP in subsequent projects. To achieve this goal, our specific aims are as follow: (1) Identify lead- or drug-like low molecular weight inhibitors of PS using a search for close analogs of the natural substrates of PS, docking of the databases of commercially available chemical compounds, de novo/rational drug design, virtual focused combinatorial libraries approaches; to synthesize the best lead candidates and analogs of the lead compounds if they are not commercially available; (2) Assay candidate drugs for their inhibition of PS functional activity. (3) Identify compounds demonstrating significant selective activity against actively growing and/or non-replicating, persistent (NRP) Mycobacterium tuberculosis. Prioritize on the basis of activity, cytotoxicity and resultant selectivity indices.