Use of Beta-lapachone for Lung Cancer Chemotherapy

描述(由申请人提供)：目前迫切需要开发一种专门治疗非小细胞肺癌患者的药物，这种患者占所有肺癌的80%，其5年生存率仅为~15%。在之前的资助期间，我们发现，在80%的非小细胞肺癌组织中，NAD(P)H：QO1的水平不仅比相关的正常组织高5-40倍，而且过氧化氢酶的水平相对地相反，在正常组织和肿瘤组织中升高。因此，NQO1是治疗消除非小细胞肺癌的理想靶点。使用NQO1的“生物活性”药物，如生成过氧化氢以激活PARP1和选择性杀死肿瘤的脱氧苯二酚(DNQ)，是实现这一策略的非常有吸引力的药物。此前，我们生成了纳米胶束，与正常组织相比，纳米粒子胶束可以有效地向非小细胞肺癌或非小细胞肺癌的正常组织输送前药，从而在原位非小细胞肺癌和其他癌症模型中产生显著的“明显治愈”的抗肿瘤反应。我们通过PARP1过度激活机制证明了NSCLC以及其他实体肿瘤的显著放射增敏作用，该机制允许使用6-10倍的低剂量和无毒剂量的电离辐射(IR)来治疗原位NSCLC异种移植。了解NQO1生物活性药物造成DNA损伤和细胞死亡的确切机制，可以提出以下新的假设和方法：我们假设亚致死剂量的LAP或DNQ87可以用来诱导肿瘤选择性的、NQO1依赖的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，产生协同抗肿瘤效应。肿瘤对DNA修复抑制剂的特异性将导致NAD+/ATP的显著丢失，以及葡萄糖代谢和DNA修复的抑制。这一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、单个或DSB修复途径以及葡萄糖代谢在亚致死剂量(NQO1+NSCLC)或DNQ87剂量(YRs)后恢复(抵抗)中的作用。1-5)。SA2：用或不用DNA修复抑制剂，和/或用或不用IR治疗(年)，以确定？-LAP-DC3-胶束或HP？CD-DNQ87的抗肿瘤效果。1-5)。两种不同的可行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ç-LAP或DNQ87来增强PARP1过度活跃的特定DNA损伤，DNQ87是我们通过抑制BER或DSB修复而开发的一种新的DNQ衍生物。在第二种方法中，PARP1将被临床相关的抑制剂阻断，这些抑制剂阻止DNA修复，并将增加NQO1生物活性药物的致命性。这两种方法都利用了NQO1生物活性药物诱导特定的肿瘤选择性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。在任何一种情况下，使用NQO1生物活性药物都将使DNA修复抑制剂具有肿瘤选择性，由于缺乏特异性，其疗效一直受到限制。

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.