Novel High Affinity VEGF Analogs For Targeted Imaging of Thyroid Cancer

用于甲状腺癌靶向成像的新型高亲和力 VEGF 类似物

基本信息

批准号：
8781160
负责人：
MARIUSZ W SZKUDLINSKI
金额：
$ 82.51万
依托单位：
TROPHOGEN, INC.
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-10 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8781160
关键词：
Affect Affinity Arginine Binding Bioreactors Cancer Control Cancer Etiology Cancer cell line Carbohydrates Cell Proliferation Cells Charge Chinese Hamster Ovary Cell Complex Coupled Coupling Data Diagnosis Diagnostic Diagnostic Imaging Diagnostic Procedure Diagnostic tests Drug Kinetics Early Diagnosis Electrostatics Endocrine Engineering Epithelium Experimental Designs Follicular thyroid carcinoma Future Gamma Cameras Glycoproteins Growth Health Hormones Human Human Chorionic Gonadotropin Human Follicle Stimulating Hormone Image In Vitro Incidence Individual Iodine Label Lead Legal patent Ligands Lipids Lysine Malignant Neoplasms Malignant neoplasm of thyroid Mammalian Cell Medical Methods Morbidity - disease rate Mutagenesis Nanotechnology Neoplasm Metastasis Normal Cell Organ Outcome Patients Peripheral Phase Phosphotransferases Plasma Prevalence Property Quality of life Radio Radioisotopes Recombinants Recurrent tumor Reproducibility Residual Tumors Roller Bottle Scanning Sensitivity and Specificity Serum Serum Markers Site Small Business Innovation Research Grant Specificity Staging Thyroglobulin Thyroid Gland Thyrotropin Receptor Time Tissues Tracer Tumor-Derived Undifferentiated Vascular Endothelial Cell Vascular Endothelial Growth Factor A Vascular Endothelial Growth Factor Receptor Vascular Endothelial Growth Factor Receptor-2 Vascular Endothelial Growth Factors Vascular Endothelium Woman Xenograft procedure analog anaplastic thyroid cancer arginyllysine base cancer cell cancer imaging cancer therapy commercialization cost design glycosylation improved in vivo in vivo imaging lysine analog mortality nanoparticle neoplastic cell novel particle receptor receptor binding receptor expression response sialylation symporter tumor tumor xenograft uptake

项目摘要

DESCRIPTION (provided by applicant): Thyroid cancer of follicular origin is the most common malignancy of endocrine tissues, disproportionally affecting women, and is one of the few cancers greatly increasing in incidence & prevalence for unknown reasons not solely attributable to improved diagnosis. Most patients require lifelong diagnostic surveillance with radioiodine imaging to detect residual & recurrent tumor requiring subsequent therapy with 131I. One of the PIs co-invented & co-developed Genzyme's recombinant human TSH (Thyrogen), currently approved for enhancing diagnostic imaging with radioiodine and for stimulation of the thyroid cancer serum marker, thyroglobulin (TG). However there is currently no optimal method to image the greatly increasing number of more aggressive cancers causing major morbidity and mortality, detected by increased serum Tg, but which have lost the ability to concentrate radioiodine because of decreased expression of the Na/I symporter. Nor is there any diagnostic method to predict which tumors may respond to increasingly important anti-angiogenic therapy for these usually highly angiogenic tumors. The two PIs have invented completely novel, patented methods to design and produce much higher affinity and longer acting analogs of VEGF which can be used in novel, targeted imaging of such tumors to the VEGFR2 receptor in both the stromal vascular endothelium and in tumor epithelium, as supported by extensive and compelling preliminary data. Two approaches of targeted imaging will be compared: a direct optimized 99mTcm labeling method of such soluble VEGF analogs and another method using novel pegylated & VEGF analog targeted "stealth" nanoparticles which can selectively transverse the highly fenestrated, leaky tumor vessels of angiogenic tumor vessels but not normal vessels. In Phase 1 of this SBIR fast track the PIs will: (1) Develop higher affinity and longer acting hVEGF analogs by lysine and arginine scanning mutagenesis of selected loops in both poles of the dimeric ligand without and with introduction of a novel neoglycosylation site adding two complex, sialylated carbohydrate chains; assessing binding of each analog to VEGFR2 and bioactivity in the stimulation of human vascular endothelial cell (HUVEC) proliferation; produce and purify 5-10 mg amounts of the final 3 glycosylated and optimally sialylated analogs in CHO cells in roller bottles; (2) Develop novel pegylated polymeric "stealth" nanoparticles targeted by covalently coupled high affinity VEGF ligand and labeled with coupled radionuclides to high specific activity; (3) Assess in vitro binding and internalization of as wellas in vivo pharmacokinetic properties of VEGF analogs directly coupled to 99mTc or 125I compared to VEGF analog-targeted labeled nanoparticles. In Phase 2 the PIs will: (1) Produce and purify large amounts (50-200 mg) of the final 2 selected VEGF analog targeting candidates in a large mammalian cell bioreactor; (2) Greatly extend Phase 1 in vivo and ex vivo imaging results by further optimizing imaging sensitivity and specificity; (3) Perform parallel quantitativ organ uptake studies with labeled VEGF analogs; (4) Make the final selection of the specific analog, radionuclide and direct labeling versus indirect nanoparticle labeling method to bring forward for commercialization based on multiple criteria. These completely novel methods for earlier detection & localization of the increasing numbers of aggressive thyroid cancers should lead to earlier and more personalized therapy predicting which patients will most benefit from increasingly important but potentially toxic anti-angiogenic therapy.

描述（由申请人提供）：滤泡起源的甲状腺癌是内分泌组织最常见的恶性肿瘤，对女性的影响尤为严重，并且是少数几种发病率和患病率大幅增加的癌症之一，原因不明，而不仅仅是由于诊断的改进。大多数患者需要使用放射性碘成像进行终生诊断监测，以检测需要后续 131I 治疗的残留和复发肿瘤。其中一位 PI 共同发明和开发了 Genzyme 的重组人 TSH (Thyrogen)，目前已被批准用于增强放射性碘诊断成像以及刺激甲状腺癌血清标记物甲状腺球蛋白 (TG)。然而，目前还没有最佳的方法来对数量急剧增加的更具侵袭性的癌症进行成像，这些癌症导致了主要的发病率和死亡率，通过血清 Tg 的增加检测到，但由于 Na/I 同向转运蛋白的表达减少，这些癌症失去了浓缩放射性碘的能力。对于这些通常高度血管生成的肿瘤，也没有任何诊断方法来预测哪些肿瘤可能对日益重要的抗血管生成治疗产生反应。两位 PI 发明了全新的专利方法，用于设计和生产亲和力更高、作用时间更长的 VEGF 类似物，可用于此类肿瘤对基质血管内皮和肿瘤上皮中 VEGFR2 受体的新型靶向成像。得到广泛且令人信服的初步数据的支持。将比较两种靶向成像方法：一种直接优化此类可溶性 VEGF 类似物的 99mTcm 标记方法，另一种方法使用新型聚乙二醇化和 VEGF 类似物靶向“隐形”纳米颗粒，该纳米颗粒可以选择性地穿过血管生成肿瘤血管的高度开窗、渗漏的肿瘤血管，但不是正常的血管。在该 SBIR 快速通道的第 1 阶段，PI 将：（1）通过对二聚配体两极中选定环的赖氨酸和精氨酸扫描诱变，开发更高亲和力和更长作用的 hVEGF 类似物，无论是否引入新的新糖基化位点，添加两个复杂的唾液酸化碳水化合物链；评估每种类似物与 VEGFR2 的结合以及刺激人血管内皮细胞 (HUVEC) 增殖的生物活性；在滚瓶中的 CHO 细胞中生产并纯化 5-10 mg 量的最终 3 种糖基化和最佳唾液酸化类似物； (2) 开发新型聚乙二醇化聚合物“隐形”纳米颗粒，以共价偶联的高亲和力 VEGF 配体为靶向，并用偶联的放射性核素标记以实现高比活性； (3) 与 VEGF 类似物靶向标记纳米颗粒相比，评估直接偶联 99mTc 或 125I 的 VEGF 类似物的体外结合和内化以及体内药代动力学特性。在第 2 阶段，PI 将： (1) 在大型哺乳动物细胞生物反应器中生产并纯化大量（50-200 mg）最终选定的 2 种 VEGF 类似物靶向候选物；（2）通过进一步优化成像灵敏度和特异性，大幅扩展一期体内和离体成像结果； (3) 用标记的 VEGF 类似物进行平行定量器官摄取研究；（4）基于多重标准，最终选择特定的类似物、放射性核素以及直接标记与间接纳米颗粒标记方法，以提出商业化。这些用于早期检测和定位越来越多的侵袭性甲状腺癌的全新方法应该会导致更早和更个性化的治疗，预测哪些患者将从日益重要但具有潜在毒性的抗血管生成治疗中最受益。