Inhibition of the ALK Receptor Kinase

描述（由申请人提供）：分泌的生长因子多效生长因子（PTN）作用于内皮细胞以及上皮细胞和成纤维细胞，并且它可以驱动肿瘤生长、血管生成、侵袭以及转移。 最近，我们鉴定了PTN的受体，孤儿酪氨酸激酶间变性淋巴瘤激酶，ALK，其激酶结构域与胰岛素受体家族具有同源性。 ALK和PTN在大多数人类癌症样本中过表达，而一系列正常组织显示未检测到受体表达。 核酶介导的胶质母细胞瘤和其他肿瘤细胞中ALK的减少导致小鼠异种移植肿瘤生长的“基因剂量”依赖性减少，并支持ALK作为药物发现的有效靶点。 最近，我们确定了在纳摩尔浓度下抑制培养细胞中PTN刺激的ALK激酶的小分子药物，并且我们使用同源建模来生成具有这些候选药物的蛋白质的3D模型。 我们假设有可能鉴定新型选择性抑制剂，并提出以下目标：目标1，先导化合物鉴定：我们将使用从头药物设计、文库筛选和虚拟组合文库生成，通过计算机模拟鉴定新型ALK激酶抑制剂。 将合成最佳候选物用于生物测定。 目的2，研究这些候选抑制剂对PTN刺激的ALK激酶活性的抑制作用，并与其他生长因子诱导的受体激酶活性进行比较。 使用这些数据的其他分子建模将用于设计和优化抑制剂。 目标3，铅优化：通过进一步完善药物/蛋白质相互作用的分子建模，改善生物测定中产生的先导化合物。 然后将在完整细胞中测定这些新抑制剂对PTN刺激的ALK激酶活性的影响。 目的4：研究最佳细胞生长抑制剂的效果和效力。 将检测选定的PTN刺激的ALK激酶抑制剂与其阻断其他生长因子相比阻断PTN诱导的细胞增殖、抗凋亡和软琼脂集落形成的能力。 总之，我们提出了基于结构的PTN/ALK信号传导抑制剂的设计和研究，作为生物学和分子建模/化学实验室之间的密切合作，目的是发现新的抗癌药物。

DESCRIPTION (provided by applicant): The secreted growth factor pleiotrophin (PTN) acts on endothelial as well as epithelial cells and fibroblasts and it can drive tumor growth, angiogenesis, invasion as well as metastasis. Recently, we identified a receptor for PTN, the orphan tyrosine kinase anaplastic lymphoma kinase, ALK that shares homology of its kinase domain with the insulin receptor family. ALK and PTN are overexpressed in a majority of human cancers samples whilst a series of normal tissues showed no detectable receptor expression. Ribozyme-mediated reduction of ALK in glioblastoma and other tumor cells resulted in a "gene dose"-dependent reduction of xenograft tumor growth in mice and supports ALK as a valid target for drug discovery. Recently we identified small-molecule drugs that inhibit the PTN-stimulated ALK kinase in cultured cells at nanomolar concentrations and we used homology modeling to generate a 3D model of the protein with these drug candidates. We hypothesize that it will be possible to identify novel selective inhibitors and propose the following aims: Aim 1, Lead identification: We will identify in silico novel ALK kinase inhibitors using de novo drug design, library screening and virtual combinatorial library generation. The best candidates will be synthesized for the biologic assays. Aim 2, to study the inhibition of PTN-stimulated ALK kinase activity in comparison to other growth factor-induced receptor kinase activity by these candidate inhibitors. Additional molecular modeling using these data will then be used to design and optimize inhibitors. Aim 3, Lead optimization: To improve lead compounds emanating from the biologic assays by further refinement of the molecular modeling of drug / protein interaction. These new inhibitors will then be assayed against PTN-stimulated ALK kinase activity in intact cells. Aim 4: To study the efficacy and potency of the best inhibitors on cell growth. Selected inhibitors of the PTN-stimulated ALK kinase will be tested for their ability to block PTN-induced cell proliferation, anti-apoptosis and soft agar colony formation in comparison to their blockade of other growth factors. In summary, we propose the design and study of structure-based inhibitors of the PTN/ALK signaling as a close collaboration between a biology and a molecular modeling/chemistry laboratory with the purpose of discovering novel anticancer drugs.