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患者占所有肺癌的约80%，其5年生存率仅为约15%。在之前的资助期内，我们发现不仅NAD(P)H：醌氧化还原酶1 （NQO1）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在正常组织与肿瘤组织中相反表达。因此，NQO1是治疗性消除非小细胞肺癌的理想靶点。使用nq01“生物激活”药物，如ß-lapachone （ß-lap）和脱氧波醌（DNQ），它们产生过氧化氢，作为一种超激活PARP1和选择性杀死肿瘤的机制，是实现这种策略的非常有吸引力的药物。在此之前，我们制作了纳米颗粒胶束，有效地将ß-lap或ß-lap前药递送到非小细胞肺癌而不是正常组织，在原位非小细胞肺癌和其他癌症模型中产生了显著的“明显治愈”的抗肿瘤反应。我们通过PARP1超激活机制证明了NSCLC以及其他实体癌的显著放射增敏，该机制允许在原位NSCLC异种移植物中使用6- 10倍降低ß-lap剂量与无毒剂量的电离辐射（IR）联合使用。了解由NQO1生物激活药物引起的DNA损伤和细胞死亡的确切机制，允许以下新的假设和方法：我们假设亚致死的ß-lap或DNQ87剂量可用于引发肿瘤选择性的NQO1依赖性DNA损伤。抑制特异性DNA碱基切除（BER）或双链断裂（DSB）修复过程将选择性地抑制NQO1+ NSCLC细胞的修复/恢复反应，从而产生协同抗肿瘤作用。肿瘤对DNA修复抑制剂的特异性将导致显著的NAD+/ATP损失和葡萄糖代谢和DNA修复的抑制。该理论将通过完成以下特定目标（SAs）来验证：SA1：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DNQ87剂量（1-5年）后NQO1+ NSCLC细胞恢复（抗性）中的作用。SA2：测定ß-lap- dc3胶束或HPßCD-DNQ87在加或不加DNA修复抑制剂和/或加或不加IR治疗时的抗肿瘤疗效（1- 5年）。两种不同可行的抗肿瘤方法将在体外（Aim 1）和体内（Aim 2）进行测试。第一种方法将使用无毒剂量的ß-lap或DNQ87（一种由我们通过抑制BER或DSB修复而开发的新型DNQ衍生物）来增强PARP1过度活跃的特定DNA病变。在第二种方法中，PARP1将被临床相关抑制剂阻断，这些抑制剂可以阻止DNA修复，并增加NQO1生物活化药物的致死率。这两种方法都利用了NQO1生物激活药物引发特异性肿瘤选择性DNA损伤的能力，但应该导致两种完全不同的细胞死亡机制：第一种策略是程序性坏死，而第二种策略应该导致经典的细胞凋亡/衰老。在这两种情况下，使用NQO1生物激活药物将使DNA修复抑制剂具有肿瘤选择性，由于缺乏特异性，其疗效受到限制。

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.