Development of (R,S')-MNF as a dual-targeted therapy for pancreatic cancer

总结 胰腺导管腺癌（PDAC）是美国癌症死亡的第三大原因。 5年生存率<9%预后不良部分是由于对标准 包括吉西他滨（Gem）和Gem+ nab-紫杉醇（n-PTX）的护理治疗。的 转录因子HIF-1α和c-MYC是Gem产生机制的中心， n-PTX抗性，并且是关键的治疗靶标。（R，S′）-4′-甲氧基-1-萘基非诺特罗（R，S ′）- MNF是一种双功能抗癌剂，其作为GPR 55的竞争性抑制剂和GPR 55的竞争性抑制剂。 β2-肾上腺素能受体的偏性激动剂。在PANC-1异种移植肿瘤模型（R，S '）-MNF中， 显著抑制肿瘤生长，抑制75%（p<0.01），并下调HIF-1α和c-MYC 表情我们的总体假设是，（R，S′）-MNF将减少PDAC肿瘤生长， 并且与标准护理剂产生积极的协同作用。整体 目的是确定（R，S′）-MNF与GEM+n-PTX联合治疗的治疗潜力， PDAC模型。实验方案将利用我们对（R，S′）-MNF的了解 药物动力学和毒性以及在PDAC模型中使用治疗剂的经验。 目的1：测定（R，S′）-MNF单独和联合应用的抗肿瘤活性。 PDAC患者来源的异种移植物（PDX）模型中的联合给药：初始步骤将是给药 寻找研究，以确定（R，S′）-MNF单独和联合的最大耐受剂量 GEM（70 mg/kg，i.p.，每周一次，持续3周）+ n-PTX（30 mg/kg，每周一次，持续3周）。 将在源自PDAC患者肿瘤的2个PDX模型中使用最佳剂量和方案 表达高水平的GPR 55和β2-AR。每项研究将包括4个治疗组，每组18人 小鼠/组：溶媒、单独的（R，S′）-MNF、GEM+n-PTX和（R，S′）-MNF + GEM+n-PTX。血液和 将从6只小鼠/组收集主要器官组织，用于目的2中提出的分析。的 监测剩余的小鼠（12只/组）以确定对肿瘤生长和存活的影响。 目的2：确定治疗生物标志物并确定（R，S′）-MNF生物分布：血浆 将使用LC-MS/MS分析治疗前后采集的样本， 分子如溶血磷脂酰胆碱（14：0和16：0）和乳酸盐，以及用于 PDAC生物标志物，如CA 19 -9和CYR 61。代谢物和蛋白质浓度将 与肿瘤生长和存活数据以及GPR 55和β2-AR的相对表达相比。 将使用LC-MS/MS分析从（R，S′）-MNF处理组采集的主要器官， 确定药物生物分布。I期研究的数据将支持I期IND研究 II申请，包括GLP PK/PD、代谢和毒性研究。

Summary Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of cancer death in the U.S. with a 5-year survival rate of <9%. The poor prognosis is partially due to resistance to standard of care treatments including gemcitabine (Gem) and Gem+nab-paclitaxel (n-PTX). The transcription factors HIF-1α and c-MYC are at the center of the mechanisms producing Gem and n-PTX resistance and are key therapeutic targets. (R,S′)-4′-Methoxy-1-naphthylfenoterol (R,S’)- MNF is a bi-functional anti-cancer agent that acts as a competitive inhibitor of GPR55 and a biased-agonist of the β2-adrenergic receptor. In a PANC-1 xenograft tumor model (R,S’)-MNF significantly dampens tumor growth, ∼75% (p<0.01), and downregulates HIF-1α and c-MYC expression. Our overarching hypothesis is that (R,S′)-MNF will reduce PDAC tumor growth as a single agent and produce positive synergistic effects with standard of care agents. The overall goal is to determine the therapeutic potential of (R,S′)-MNF in combination with GEM+n-PTX in PDAC models. The experimental protocols will utilize our knowledge of (R,S′)-MNF pharmacokinetics and toxicity and experience with use of therapeutic agents in PDAC models. Specific aims are: Aim 1: to determine the antitumor activity of (R,S′)-MNF alone and in combination in PDAC patient derived xenograft (PDX) models: The initial step will be a dosing finding study to determine maximal tolerated dose of (R,S′)-MNF alone and in combination with GEM (70 mg/kg, i.p., once a week for 3 weeks) + n-PTX (30 mg/kg, once a week for 3 weeks). Optimal dose and schedule will be used in 2 PDX models derived from PDAC patients’ tumors expressing high levels of GPR55 and β2-AR. Each study will include 4 treatment groups of 18 mice/group: vehicle, (R,S′)-MNF alone, GEM+n-PTX, and (R,S′)-MNF + GEM+n-PTX. Blood and major organ tissues will be collected from 6 mice/group for analyses proposed in Aim 2. The remaining mice (12/group) will be monitored to determine the effect on tumor growth and survival. Aim 2: to identify treatment biomarkers and determine (R,S′)-MNF biodistributions: Plasma samples collected before and after treatment will be analyzed using LC-MS/MS to quantify small molecules such as lysophosphatidylcholines (14:0 and16:0) and lactate and ELISA assays for PDAC biomarkers such as CA19-9 and CYR61. Metabolite and protein concentrations will be compared to tumor growth and survival data and to the relative expression of GPR55 and β2-AR. Major organs collected from the (R,S′)-MNF treatment group will be analyzed using LC-MS/MS to determine drug biodistribution. Data from the Phase I study will support IND studies in a Phase II application, which will include GLP PK/PD, metabolism and toxicity studies.