New Stereoregular Functionalized Fullerenes as Nanomedicines for PDT

描述(由申请人提供)：本申请的目标是(I)设计和合成光敏性[60]富勒烯对映体和相关纳米结构作为光敏PDT应用，以及(Ii)开展两亲性多功能[60]富勒烯对映体和发色团纳米结构作为1PA-PDT和2PA-PDT(2-3吸收)纳米药物的体外和体内生物研究。我们建议设计和合成新型阳离子立体规则1-吡咯烷基[60]富勒烯对映体PF-PhOEGn和阳离子C60(&GT；CPAF-EG6)x作为光动力治疗药物，具有以下特点：(1)只需一次给药即可对多种PDT治疗具有高的光稳定性；(2)利用C60天线结构增强光捕获能力，用于细胞毒性ROS的产生；(3)C60天线的分子自组装能力，形成稳定的囊泡，用于PDT药物的大容量输送；(4)双光子吸收截面[&GT；4500 GM用于C60-(天线)2]用于有效的2PA-PDT治疗。我们已经确定了富勒烯的一些分子结构特征，它们介导了对微生物细胞和癌细胞的广谱光杀灭。这种对所需结构基序的洞察已被应用于富勒烯纳米药物的分子设计。我们将通过体外和体内实验来研究PDT的效率：(1)我们已经组装了一组病原微生物，旨在涵盖大多数引起局部感染的微生物，例如伤口和烧伤，这些微生物表现出内在和获得性多重抗生素耐药性。聚阳离子富勒烯将作为PDT试剂进行测试，以对抗这些病原体。(2)对于富勒烯介导的PDT治疗局部感染，我们将使用基因工程生物发光细菌结合光学成像来研究创面感染的小鼠模型，以实时无创地跟踪感染的进展。(3)我们将在体外对一组小鼠癌细胞进行光动力疗法，预计这些细胞系对富勒烯的摄取不同，对光动力疗法产生的活性氧物种的细胞反应也不同。药物和光的剂量效应研究将在单光子和双光子激发下进行，并研究细胞死亡的机制。(4)为了抗癌测试，我们将使用小鼠皮下肿瘤，并比较瘤内注射和静脉注射富勒烯的效果。我们将比较KTP激光的单光子激发和飞秒Ti：Sapph激光的双光子激发。 公共卫生相关性：本申请中描述的富勒烯纳米药物有可能中介对当代两种主要致命疾病的光动力疗法。通过适当的富勒烯分子设计和连续波或飞秒脉冲光的照射相结合，多药耐药细菌感染和癌症肿瘤可以被消除。本文提出的这些化合物的特殊化学和光物理特征将改善这些疾病的光动力疗法，使之超越目前可能的水平。

DESCRIPTION (provided by applicant): The objective of this application is to (I) design and synthesize photoresponsive [60]fullerenyl enantiomers and related nanostructures as photosensitizers for PDT applications and (ii) carry out in vitro and in vivo biostudies of amphiphilic multifunctional [60]fullerenyl enantiomers and chromophore nanostructures as 1PA-PDT and 2PA-PDT (2-3 absorptive) nanomedicines. We propose to design and synthesize novel cationic stereoregular 1-pyrrolino[60]fullerenyl enantiomers PF-PhOEGn and cationic C60(>CPAF-EG6)x for the use as PDT agents with the following characteristics: (1) high photo stability for multiple PDT treatments with only a single-dose of the drug; (2) enhancement of light-harvesting capability using the C60-antenna construction, for cytotoxic ROS production; (3) capability of molecular self-assembly of C60-antenna forming stable vesicles for high volume delivery of PDT drug; (4) large enhancement of two-photon absorption cross-sections [>4500 GM for C60- (antenna)2] for effective 2PA-PDT treatments. We have established some of the molecular structural features of fullerenes that mediate broad-spectrum photokilling of microbial cells and cancer cells. This insight into required structural motifs has been applied to the molecular design of fullerene nanomedicines. We will study PDT efficiency by both in vitro and in vivo experiments: (1) we have assembled a panel of pathogenic microorganisms designed to encompass most of the microbes that cause localized infections e.g. in wounds and burns, and which demonstrate both intrinsic and acquired multi-antibiotic resistance. Polycationic fullerenes will be tested as PDT agents against these pathogens. (2) For fullerene-mediated PDT treatment against localized infections we will study a mouse model of wound infection using genetically engineered bioluminescent bacteria combined with optical imaging to follow the progress of the infection non-invasively in real time. (3) We will carry out in vitro PDT against a panel of mouse cancer cell lines that are expected to have different uptakes of fullerenes and different cellular responses to reactive oxygen species produced by PDT. Dose response studies of both drug and light will be carried out for both 1-photon and 2-photon excitation and mechanisms of cell death studied. (4) For anti-cancer testing we will use subcutaneous mouse tumors and compare intratumoral injection with IV delivery of the fullerene. We will compare both 1-photon excitation using a KTP laser and 2-photon excitation using a femtosecond Ti:sapph laser. PUBLIC HEALTH RELEVANCE: The fullerene nanomedicines described in this application have the potential to mediate photodynamic therapy of two of the major killer diseases of the present age. Multi-drug resistant bacterial infections and cancerous tumors could be eliminated by a combination of the appropriate molecular design of fullerene and illumination with CW or femtosecond pulsed light. Particular chemical and photophysical features of these compounds proposed herein will improve PDT of these diseases beyond what is presently possible.