Use of Beta-lapachone for Lung Cancer Chemotherapy

描述（由申请人提供）：迫切需要开发特异性和有效治疗NSCLC患者的药物，NSCLC患者占所有肺癌的>80%，其5年生存率仅为~ 15%。在之前的资助期间，我们发现，不仅NAD（P）H：醌氧化还原酶1（NQO 1）水平在>80%的NSCLC肿瘤中比相关正常组织升高5- 40倍，而且过氧化氢酶水平相对而言呈负向表达，在正常组织中比肿瘤组织中升高。因此，NQO 1代表了用于治疗性消除NSCLC的完美靶标。使用NQO 1“生物可活化”药物，如产生过氧化氢作为过度活化PARP 1和选择性杀死肿瘤的机制的β-拉帕醌（β-lap）和脱氧尼波醌（DNQ），是实现这种策略的非常有吸引力的药物。以前，我们产生了纳米粒子胶束，其有效地将ss-lap或ss-lap前药递送到NSCLC与正常组织，导致在原位NSCLC和其他癌症模型中具有显著“明显治愈”的抗肿瘤反应。我们通过PARP 1超活化机制证明了NSCLC以及其他实体癌的显著放射增敏作用，该机制允许使用6至10倍降低的ss-lap剂量与无毒剂量的电离辐射（IR）组合用于原位NSCLC异种移植物的疗效。了解NQO 1生物活化药物引起的DNA损伤和细胞死亡的确切机制允许以下新的假设和方法：我们假设亚致死ss-lap或DNQ 87剂量可用于引发肿瘤选择性，NQO 1依赖性DNA损伤。特异性DNA碱基切除（BER）或双链断裂（DSB）修复过程的抑制将选择性抑制NQO 1 + NSCLC细胞中的修复/恢复反应，从而产生协同抗肿瘤效应。DNA修复抑制剂的肿瘤特异性将导致显著的NAD+/ATP损失以及葡萄糖代谢和DNA修复的抑制。将通过完成以下特定目的（SA）来检验该理论：SA 1：阐明特定DNA碱基、单一或DSB修复途径和葡萄糖代谢在亚致死ss-lap或DNQ 87给药后NQO 1 + NSCLC细胞恢复（耐药性）中的作用（第1-5年）。SA2：确定ss-lap-dC 3-胶束或HPssCD-DNQ 87在有或没有DNA修复抑制剂和/或有或没有IR处理的情况下的抗肿瘤功效（Yrs.1 - 5）。将在体外（目标1）和体内（目标2）测试两种不同的可行抗肿瘤方法。第一种方法将使用无毒剂量的ss-lap或DNQ 87（我们通过抑制BER或DSB修复开发的一种新型DNQ衍生物）来增强过度激活PARP 1的特定DNA损伤。在第二种方法中，PARP 1将被阻止DNA修复的临床相关抑制剂阻断，并将增加NQO 1生物活化药物的致死率。这两种方法都利用NQO 1生物活化药物引起特异性肿瘤选择性DNA损伤的能力，但应导致两种完全不同的细胞死亡机制：第一种策略中的程序性坏死，而第二种策略应导致经典的凋亡/衰老。在任何一种情况下，使用NQO 1生物可活化药物将赋予DNA修复抑制剂肿瘤选择性，其功效由于缺乏特异性而受到限制。

DESCRIPTION (provided by applicant): There is a desperate need to develop agents that specifically and efficaciously treat NSCLC patients, which represent >80% of all lung cancers and whose 5 year survival rates are only ~15%. In the prior funding period, we discovered that not only were NAD(P)H:quinone oxidoreductase 1 (NQO1) levels elevated 5- to 40-fold in >80% NSCLC tumors vs associated normal tissue, but that catalase levels were inversely expressed comparatively, elevated in normal vs tumor tissue. NQO1, therefore, represents a perfect target to exploit for the therapeutic elimination of NSCLCs. Use of NQO1 'bioactivatable' drugs, such as ss-lapachone (ss-lap) and deoxynyboquinone (DNQ) that generate hydrogen peroxide as a mechanism to hyperactivate PARP1 and selectively kill tumors, are very attractive drugs to enable such a strategy. Previously, we generated nanoparticle micelles that efficaciously delivered ss-lap or ss-lap prodrugs to NSCLCs vs normal tissue, resulting in antitumor responses with significant 'apparent cures' in orthotopic NSCLC and other cancer models. We demonstrated significant radiosensitization of NSCLCs, as well as other solid cancers, via a PARP1 hyperactivation mechanism that allowed use of 6- to 10-fold lowered ss-lap doses in combination with nontoxic doses of ionizing radiation (IR) for curative effects in orthotopic NSCLC xenografts. Understanding the exact mechanism of DNA damage and cell death caused by NQO1 bioactivatable drugs allows the following novel next hypotheses and approaches: We hypothesize that sublethal ss-lap or DNQ87 doses can be used to elicit tumor-selective, NQO1-dependent DNA damage. Inhibition of specific DNA base excision (BER) or double strand break (DSB) repair processes will selectively suppress repair/recovery responses in NQO1+ NSCLC cells, causing synergistic antitumor effects. Tumor-specificity to DNA repair inhibitors will resul in dramatic NAD+/ATP losses and inhibition of glucose metabolism and DNA repair. This theory will be tested by completing the following Specific Aims (SAs): SA1: To elucidate the roles of specific DNA base, single or DSB repair pathways and glucose metabolism in recovery (resistance) of NQO1+ NSCLC cells after sublethal ss-lap or DNQ87 doses (Yrs. 1-5). SA2: To determine the antitumor efficacy of ss-lap- dC3-micelles or HPssCD-DNQ87, with or without DNA repair inhibitors, and/or with or without IR treatments (Yrs. 1- 5). Two distinct viable antitumor approaches will be tested in vitro (Aim 1) and in vivo (Aim 2). The first approach will augment specific DNA lesions that hyperactive PARP1 using nontoxic doses of ss-lap or DNQ87, a novel DNQ derivative developed by us by inhibiting BER or DSB repair. In the second approach, PARP1 will be blocked by clinically-relevant inhibitors that prevent DNA repair and will augment lethality of NQO1 bioactivatable drugs. Both approaches exploit the ability of NQO1 bioactivatable drugs to elicit specific tumor-selective DNA lesions, but should result in two completely different cell death mechanisms: programmed necrosis in the first strategy, while the second should cause classical apoptosis/senescence. In either case, use of NQO1 bioactivatable drugs will lend tumor-selectivity to DNA repair inhibitors, whose efficacy has been limited due to lack of specificity.