Novel Tiancimycins (TNMs) Antibody Drug Conjugates (ADCs) for Anticancer Therapeutics

项目摘要 抗体-药物偶联物（ADC）领域已成为下一代靶向治疗选择 并将高细胞毒性药物递送至肿瘤。这一领域的快速增长体现在60多种ADC中 目前正在进行临床试验，2011年FDA批准了四种药物Adcetris®，2013年批准了Kadcyla®， 2017年，Besponsa®和Mylotarg®。目前，在ADC领域，有两个主要问题需要解决。 克服：（i）随机缀合和变化的药物与抗体比率（DAR），和（ii）小分子药物， 有效载荷几乎所有临床试验中的ADC和两种批准的药物都使用随机偶联来连接ADC。 有效载荷与抗体内存在的赖氨酸或半胱氨酸残基连接。这导致具有DAR的异质ADC 0到8之间。缺乏一致性导致药代动力学和安全性问题。一种克服 这是通过抗体内的特定残基的位点特异性缀合。目前，小 用作ADC有效载荷的分子是有限的，并且是新的、高效的和快速活化的小分子 是必要的。我们的假设是：（i）生产的天蚕霉素（TNM）和TNM类似物和结构-活性 （ii）开发用于TNM的位点特异性缀合的接头化学，（iii） 抗HER和抗ROR 1 thiomab-和DVD-TNM组针对HER 2 +/ROR 1-和HER 2-的评价 /ROR 1+乳腺癌细胞系用于这些新的基于烯二炔的ADC与当前的ADC的直接比较。 基准以及开发用于缺乏当前治疗方法的乳腺癌的下一代ADC 选项.本申请的具体目的是（i）通过以下步骤生产、分离和发酵TNM： 链霉菌CB 03234中TNM生物合成的操纵和化学多样化，（ii）开发 和优化用于TNM与一组抗HER 2和抗ROR 1的位点特异性缀合的接头化学 （iii）评估TNM类似物和TNM缀合物针对以下的效力和选择性： 体外HER 2 +/ROR 1-和HER 2-/ROR 1+乳腺癌细胞。此应用程序的结果包括 ADC领域的进展，包括新的烯二炔有效载荷、新的位点特异性缀合技术，以及 用于开发下一代ADC的新型抗HER 2和抗ROR 1 thiomab和DVD-TNM ADC 乳腺癌的治疗方法这项研究的长期目标是确定创新的细胞毒性天然产物 以用作ADC的小分子有效载荷，从而提供新型抗癌药物。

Project Summary The antibody-drug conjugate (ADC) field has emerged as the next-generation therapeutic option for targeting and delivering highly cytotoxic drugs to tumors. The rapid growth in this area is seen in the more than 60 ADCs currently in clinical trials and the four FDA-approved drugs Adcetris® in 2011, Kadcyla® in 2013, and most recently Besponsa®, and Mylotarg® in 2017. Currently within the ADC field there are two major issues that need to be overcome: (i) random conjugation and varying drug-to-antibody ratios (DAR), and (ii) small molecule drugs as payloads. Nearly all the ADCs in clinical trials and the two approved drugs use random conjugation linking the payload to lysine or cysteine residues present within the antibody. This results in heterogenous ADCs with DARs between 0 and 8. The lack of uniformity leads to issues in pharmacokinetics and safety. One way to overcome this is site-specific conjugation through specific residues within the antibody. Furthermore, the currently small molecules being used as payloads for ADCs is limited and new, highly potent, and rapidly active small molecules are needed. Our hypotheses are (i) production of tiancimycins (TNMs) and TNM analogs and structure-activity relationship (SAR) studies, (ii) development of linker chemistry for site-specific conjugation of the TNMs, (iii) evaluation of the panel of anti-HER and anti-ROR1 thiomab- and DVD-TNMs against HER2+/ROR1- and HER2- /ROR1+ breast cancers cell lines for direct comparison of these novel enediyne-based ADCs to current benchmarks as well as development into next-generation ADCs for breast cancer lacking current therapeutic options. The specific aims for this application are (i) production, isolation, and fermentation of TNMs via manipulation of TNM biosynthesis in Streptomyces sp. CB03234 and chemical diversification, (ii) development and optimization of linker chemistry for site-specific conjugation of TNM to a panel of anti-HER2 and anti-ROR1 thiomabs and DVDs, and (iii) evaluation of potency and selectivity of TNM analogs and TNM conjugates against HER2+/ROR1- and HER2-/ROR1+ breast cancer cells in vitro. The outcomes of this application include advancements in the ADC field including new enediyne payloads, new site-specific conjugation techniques, and new anti-HER2 and anti-ROR1 thiomab- and DVD-TNM ADCs for the development of next-generation ADC therapies for breast cancer. The long-term goal of this research is to identify innovative cytotoxic natural products to utilize as small molecule payloads for ADCs to afford novel anticancer drugs.