Treatment of Glioblastoma: the combination of fluorescence guided surgery and photodynamic therapy

摘要： 先前资助的STTR（快速通道I/II期，临床前研究）项目（R41/R42 CA 173980 - 01 A） 帮助我们开发出所需的NIR PS（Photobac），在癌症成像和治疗方面具有巨大潜力。 在GMP装置中还建立了一种有效的合成Photobac的方法。追随美国 FDA指南、不同剂量Photobac的药代动力学（PK）、药效学（PD）和毒性 还在小鼠、大鼠和狗中进行了研究。组织学结果未显示任何器官毒性。Photobac 即使在比治疗剂量高20倍的剂量（治疗所需的单剂量）下也不是致命的。没有 甚至在高于所需治疗剂量时也观察到代谢物形成和器官毒性。的 还在小鼠和大鼠中测定了C-14 Photobac在不同时间点的生物分布[7，8]。结果 在大鼠中使用C-14 Photobac和在小鼠中的荧光结果表明，Photobac穿过血脑 屏障（BBB）。 美国FDA对我们要求使用Photobac治疗胶质瘤的IND前积极回应是 令人愉悦和兴奋（申请号PIND 136905）。根据FDA审查委员会的建议， 人类患者I期方案已经完成。近红外PS Photobac还收到了“孤儿” 美国FDA授予的胶质母细胞瘤成像和治疗（荧光引导手术+ PDTof复发肿瘤）（# DRU-2017-6153），这进一步鼓励我们将这项技术转移到 I期人体临床试验。为了实现我们的目标，拟议的研究（SBIR：临床前和临床） 分为以下目标： 目的1（a）：研究Photobac在大鼠脑胶质瘤模型中的PDT疗效，并使用Simphotek的专利产品 计算装置FPDosi™（Beeson等人，J. Biomed.光学24（3），035006，2019）比较光剂量与 实验详情见Simphotek的支持信。目标1（B）：编制 Photobac CMC（化学、生产和控制）、药物配制、原料药和药物稳定性 IND的产品、分析细节、PK/PD、毒性、体外和体内数据以及I期临床方案 向美国FDA提交了胶质母细胞瘤的荧光图像引导手术+ PDT。IND的成本 提交将由Photolitec，LLC支付。 目的2：编制PS（Photobac）定制的NIR激光仪（787 nm，与 Biolitec的帮助），并提交批准（合并IND-目标1]。 目的3：使用Photobac-PDT进行I期试验，荧光引导治疗（手术+ PDT） 用于选择最佳治疗参数的单一光剂量和可变药物剂量下的胶质母细胞瘤。 目的4：提供来自胶质母细胞瘤、相应的非肿瘤组织、未治疗的肿瘤的组织样品， Photobac-PDT治疗肿瘤和既往手术、化疗、放疗和化疗后复发的肿瘤。 化疗治疗，使用3D细胞培养系统的Photobac特异性，和STAT 3交联分析。 目的5：建立植入的患者肿瘤（在不同阶段）的细胞摄取之间的相关性。 小鼠，光诱导的STAT 3二聚化和长期PDT反应[7]。把这些发现翻译成 增强胶质母细胞瘤患者的最佳治疗参数。

ABSTRACT: The previously funded STTR (Fast Track Phase I/II, preclinical studies) project (R41/R42 CA173980-01A) helped us in developing desired NIR PS (Photobac) with a great potential for both cancer imaging and therapy. An efficient method for the synthesis of Photobac was also established in a GMP facility. Following the US FDA guidelines, the pharmacokinetic (PK), pharmacodynamic (PD) and toxicity of Photobac in various doses were also investigated in mice, rats and dogs. Histopathology results did not show any organ toxicity. Photobac even at a 20-fold higher than the therapeutic dose (single dose needed for the treatment) was not lethal. No metabolite formation and organ toxicity even at higher than the desired therapeutic dose was observed. The biodistribution of C-14 Photobac at variable time points was also determined in mice and rats [7, 8]. Results with C-14 Photobac in rats and fluorescence results in mice suggest that Photobac crosses the blood brain barrier (BBB). A positive pre-IND response from the US FDA on our request for the use of Photobac to treat Gliomas was pleasing & exciting (Application # PIND 136905). As suggested by the FDA review committee, the revision of human patient Phase I protocol has been accomplished. NIR PS Photobac has also received an “Orphan Drug” status from the US FDA for imaging and therapy of Glioblastoma (fluorescence-guided surgery + PDTof recurrence tumors) (# DRU-2017-6153), which further encouraged us to move this technology to Phase I human clinical trials. To achieve our goal, the proposed study (SBIR: Preclinical and Clinical)) has been divided into following aims: Aim 1 (a): To investigate the PDT efficacy of Photobac in rat glioma model and using Simphotek’s proprietary computational device FPDosi™ (Beeson et al. J. Biomed. Opt. 24(3), 035006, 2019) compare light dose to experiments. Details are presented in the letter of support from Simphotek. Aim 1 (b): To compile the Photobac CMC (chemistry, manufacturing and control), drug-formulation, stability of drug substance and drug product, analytical details, PK/PD, toxicity, in vitro & in vivo data and Phase I clinical protocol for an IND submission to the US FDA for fluorescence image guided surgery + PDT of glioblastoma. The cost of the IND submission will be paid by Photolitec, LLC. Aim 2: To compile the details of PS (Photobac)-customized NIR laser instrument (787 nm, developed with the help of Biolitec) following the US FDA guidelines, and submit it for approval (combined IND-Aim 1]. Aim 3: To perform Phase I trial with Photobac-PDT, fluorescence-guided treatment (surgery + PDT) of glioblastoma at a single light dose and variable drug doses for selecting the best treatment parameters. Aim 4: To provide tissue samples from glioblastoma, corresponding non-tumor tissue, untreated tumors, Photobac-PDT treated tumors and recurrent tumors after previous surgery, chemotherapy, radiation and chemotherapy treatment, for Photobac specificity using 3D cell-culture system, and STAT3 crosslinks analysis. Aim 5: Establish a correlation between the cell-uptake on patients’ tumors (at various stages) implanted in mice, photo-induced STAT3 dimerization and a long-term PDT response [7]. Translate these findings to enhance the optimal treatment parameter(s) of glioblastoma patients.