DNA-modulated release of drug from melanoma targeting NP

描述（由申请人提供）：本研究的总体目标是开发新型DNA调节的药物释放纳米颗粒（DDRNP），其可以以高特异性靶向和治疗黑色素瘤或其他类型的癌症。纳米技术是一个新兴领域，已显示出开发用于各种疾病（包括癌症）的新型诊断、成像和治疗剂的前景。纳米治疗剂或纳米成像剂的有效临床应用的一个主要障碍是缺乏高亲和力和高特异性的靶向配体，其可以在体内高效地将这些纳米药物递送到肿瘤或器官靶位点。目前需要用于未来纳米医学的递送和靶向的替代方法。我们最近报道了（Peng，2006）使用多样的和高度集中的单珠单化合物（OBOC）组合肽模拟物文库结合高严格性筛选方法鉴定针对活化的1421整联蛋白的高亲和力（IC 50 =2 pM）肽模拟物配体（LLP 2A）。我们进一步证明，LLP 2A能够成像1421表达的淋巴瘤，具有高灵敏度和特异性时，共轭近红外荧光染料在小鼠异种移植模型。此外，它还与许多肿瘤类型的生长血管结合。我们还报道了卵巢癌靶向配体（OA 02）的鉴定，其以高特异性结合1321整联蛋白（Aina，2005 a）。体内光学（Aina，2005 b）和PET（Aina，2007）成像研究证实了其癌症靶向潜力。最近，我们能够证明这种配体和相关配体可以以高特异性靶向恶性黑色素瘤（见初步数据部分）。蒽环类药物是一类有效的DNA嵌入药物，用于治疗癌症，包括黑色素瘤，但其治疗效果受到其毒性和缺乏特异性的限制。我们设想，可以利用其有效的dsDNA嵌入，以开发一种新的药物输送模式，通过创建短的蒽环类负载的dsDNA序列作为药物载体，从而它们的热变性触发抗癌药物的局部释放。携带蒽环类药物负载的dsDNA序列和OA 02/LLP 2A靶向配体的磁性纳米颗粒（MNP）将被选择性地递送至癌组织。外部高频电磁场（射频或RF加热）将被MNP转换为局部热输出，导致dsDNA变性和伴随的原位药物释放。我们的假设是，DNA调节的药物释放概念与黑色素瘤靶向配体结合应用，将可用作I、II和III期黑色素瘤患者的辅助治疗，以及作为更晚期疾病患者的姑息治疗。在这项R21应用中，我们将在黑色素瘤异种移植模型中开发这种新型纳米治疗方法。本申请的具体目的如下：目的1：开发、制备和表征用OA 02（黑素瘤靶向配体）和/或LLP 2A（肿瘤血管靶向配体）修饰的DNA调节的药物释放纳米颗粒（DDRNP）。目的2：研究射频（RF）触发DDRNP中阿霉素（DOX）的体外释放及抗癌作用。目的3：研究DDRNPs在小鼠黑色素瘤移植瘤模型中的生物学分布和瘤内分布。职务名称：DNA调节的药物从黑色素瘤靶向NP项目Narrative短的载有蒽环类药物的dsDNA序列可以用作药物载体，由此它们的热变性触发抗癌药物的局部释放。携带蒽环类药物负载的dsDNA序列和OA 02/LLP 2A靶向配体的磁性纳米颗粒（MNP）将被选择性地递送至癌组织。外部高频电磁场（射频或RF加热）将被MNP转换为局部热输出，导致dsDNA变性和伴随的原位药物释放。我们的假设是，DNA调节的药物释放概念与黑色素瘤靶向配体结合应用，将可用作I、II和III期黑色素瘤患者的辅助治疗，以及作为更晚期疾病患者的姑息治疗。在该R21申请中，我们将在黑素瘤异种移植模型中开发这种新的纳米方法。

DESCRIPTION (provided by applicant): The overall goal of this research is to develop novel DNA-modulated drug release nanoparticles (DDRNP) that can target and treat melanoma, or other type of cancer, with high specificity. Nanotechnology is an emerging field that has shown promise for the development of novel diagnostic, imaging and therapeutic agents for a variety of diseases, including cancer. One major obstacle to the effective clinical application of nano-therapeutic or nano-imaging agents is the lack of high affinity and high specificity targeting ligands that can deliver these nanomedicines to the tumor or organ target site with high efficiency in vivo. There are presently needs for alternative methods of delivery and targeting for future nanomedicines. We have recently reported (Peng, 2006) the identification of a high-affinity (IC50=2 pM) peptidomimetic ligand (LLP2A) against activated 1421 integrin using both diverse and highly focused one-bead one-compound (OBOC) combinatorial peptidomimetic libraries in conjunction with a high stringency screening method. We further demonstrated that LLP2A is able to image 1421- expressing lymphomas with high sensitivity and specificity when conjugated to a near infrared fluorescent dye in a murine xenograft model. In addition, it also binds to the growing blood vessels of many tumor types. We have also reported the identification of an ovarian cancer targeting ligand (OA02) that bind to 1321 integrin with high specificity (Aina, 2005a). In vivo optical (Aina, 2005b) and PET (Aina, 2007) imaging studies have confirmed its cancer targeting potential. Very recently, we were able to demonstrate that this and related ligands can target malignant melanoma with high specificity (see Preliminary Data section). Anthracyclines are a class of potent DNA intercalating drugs used for treatment of cancer, including melanoma, but their therapeutic efficacy is limited by their toxicity and lack of specificity. We are envisaging that their efficient dsDNA intercalation can be exploited to develop a new drug delivery paradigm by creating short anthracycline-laden dsDNA sequences as drug carriers whereby their thermal denaturation triggers the localized release of the anticancer drug. Magnetic nanoparticles (MNPs) carrying both anthracycline-loaded dsDNA sequences and OA02/LLP2A targeting ligands will be delivered selectively to cancer tissues. An external high frequency electromagnetic field (radiofrequency or RF heating) will be transduced by the MNP to a localized thermal output causing dsDNA denaturation and concomitant in situ drug release. Our hypothesis is that the DNA-modulated drug release concept applied in conjunction to melanoma targeting ligands, will be useful as adjuvant therapy for stage I, II, and III melanoma patients, and as palliative therapy for patients with more advanced disease. In this R21 application we shall develop this novel nanotherapeutic approach in a melanoma xenograft model. The specific aims of this application are as follows: Aim 1: To develop, prepare, and characterize DNA-modulated drug release nanoparticles (DDRNP) decorated with OA02 (a melanoma targeting ligand), and/or LLP2A (a tumor blood vessel targeting ligand). Aim 2: To evaluate the RF triggered in vitro release and anti-cancer effects of doxorubicin (DOX) from DDRNP. Aim 3: To evaluate the biodistribution and intratumoral distribution of DDRNPs in the murine xenograft model for melanoma. Title: DNA-modulated release of drug from melanoma targeting NP Project Narrative Short anthracycline-laden dsDNA sequences can be used as drug carriers whereby their thermal denaturation triggers the localized release of the anticancer drug. Magnetic nanoparticles (MNPs) carrying both anthracycline-loaded dsDNA sequences and OA02/LLP2A targeting ligands will be delivered selectively to cancer tissues. An external high frequency electromagnetic field (radiofrequency or RF heating) will be transduced by the MNP to a localized thermal output causing dsDNA denaturation and concomitant in situ drug release. Our hypothesis is that the DNA-modulated drug release concept applied in conjunction to melanoma targeting ligands, will be useful as adjuvant therapy for stage I, II, and III melanoma patients, and as palliative therapy for patients with more advanced disease. In this R21 application we shall develop this novel nanotherapeutic approach in a melanoma xenograft model.