Novel Chemotherapeutics for Brain Tumors

脑肿瘤的新型化疗药物

基本信息

批准号：
8523398
负责人：
Bernd Jandeleit
金额：
$ 28.81万
依托单位：
QUADRIGA BIOSCIENCES, 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：
2013-06-01 至 2014-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8523398
关键词：
Address Affect Affinity Alkylating Agents Amino Acid Transport System L Amino Acid Transporter Amino Acids Antineoplastic Agents Applications Grants Binding Biological Assay Blood - brain barrier anatomy Blood Circulation Bone Marrow Bone Marrow Suppression Brain Brain Neoplasms Cancer cell line Cancerous Capillary Endothelial Cell Cell Line Cell Proliferation Cell membrane Cells Coupled Cytotoxic agent Cytotoxin Data Detection Development Diagnosis Diffuse Diffusion Disease Dose-Limiting Drug Delivery Systems Elements Endothelial Cells Engineering Erythroid Progenitor Cells Essential Amino Acids Evaluation Exhibits Funding Goals Hybrids In Vitro Incidence Lethal Dose 50 Leucine Lipids Lomustine Malignant Neoplasms Malignant neoplasm of brain Mean Survival Times Mediating Melphalan Membrane Methods Mission Modeling Myelogenous N Domain Neuraxis Nitrosourea Compounds Normal tissue morphology Nutrient Organ Patients Penetration Permeability Pharmaceutical Preparations Phase Phenylalanine Property Reagent Research Side Small Business Innovation Research Grant Specificity Stem cells Testing Therapeutic Tight Junctions Tissues Toxic effect Transmembrane Transport United States National Institutes of Health Xenograft Model base cancer cell cerebrovascular chemotherapeutic agent chemotherapy cytotoxic cytotoxicity gabapentin gamma-Aminobutyric Acid improved in vitro testing in vivo inhibitor/antagonist liquid chromatography mass spectrometry monolayer neoplastic cell novel outcome forecast phase 2 study pregabalin public health relevance receptor research study solute stereochemistry success tumor tumor growth uptake

项目摘要

DESCRIPTION (provided by applicant): Following systemic administration, most anticancer drugs exhibit very limited uptake into the brain. The blood- brain barrier (BBB) is the single most important factor for restricting drug delivery to the brain. The BBB is formed by a continuous monolayer of cerebrovascular endothelial cells joined together by tight junctions and lacks fenestrations. To cross the BBB, solutes must diffuse across the lipid endothelial cell membrane or be carried across either by receptor-mediated transport or by membrane transport. The capillary endothelial cells of the BBB express an array of uptake and efflux transporters. Regulated expression and function of these transporters govern access of essential nutrients and therapeutic drugs to the central nervous system (CNS). Successful drug delivery to brain tumors includes not only penetration of the BBB, but also delivery to and uptake into tumor cells. The Large Amino Acid Transporter 1 (LAT1) fulfills all necessary requirements - expression at both sides of the BBB, high expression in almost all tumor cells regardless of the tissue of origin, and very low endogenous expression in non-proliferative cells. Furthermore, it is characterized by an appreciable capacity, which can be exploited to deliver therapeutically meaningful amounts of chemotherapeutic agents. LAT1 activity correlates with cell proliferation, tumor growth, and is inversely related to disease prognosis and patient survival. Quadriga BioSciences will use NIH/NCI funding to engineer novel anti-cancer agents by combining the properties of cytotoxic agents with LAT1-mediated delivery to cancerous cells. The aims of Phase 1 of the proposed SBIR research are geared to determine whether LAT1-transported anticancer agents are a viable option for the development of chemotherapeutics to treat brain tumors. Success of our strategy is defined as activity and high specificity for LAT1. In Aim 1 a set of compounds will be synthesized that are "hybrid" compounds in which a known cytotoxic moiety is chemically fused to a LAT1- specific recognition element. These compounds will be tested for LAT1 activity and specificity in Aim 2. LAT1 transport will be determined in vitro using [3H]-gabapentin uptake competition in HEK cells and direct uptake coupled with LC/MS/MS detection. The specificity for LAT1 vs. other amino acid transporters will be tested by [3H]-phenylalanine uptake competition in cells expressing those transporters. Possible BBB transport of the compounds will be assessed by and in vitro transwell transport assay. In Aim 3, cytotoxicity (LD50 and LD100) of compounds toward cancer cell lines and bone marrow progenitor cells is evaluated. To exclude that test compounds enter the cell via means other than LAT1, the effect of LAT1 inhibitors on the cytotoxicity will be determined. A successful compound will be cytotoxic to LAT1-expressing cancer cell lines through selective accumulation and exhibit no cytotoxicity to bone marrow progenitor cells. In projected Phase 2 studies, successful compounds will be tested in vivo in validated xenograft models for various brain tumors.

描述（由申请人提供）：全身给药后，大多数抗癌药物在大脑中的吸收非常有限。血脑屏障（BBB）是唯一最限制药物向大脑输送的重要因素。 BBB 由连续的单层脑血管内皮细胞通过紧密连接连接在一起形成，并且没有开窗。为了穿过血脑屏障，溶质必须扩散穿过脂质内皮细胞膜，或者通过受体介导的转运或膜转运被携带穿过。 BBB 的毛细血管内皮细胞表达一系列摄取和流出转运蛋白。这些转运蛋白的表达和功能的调节控制着必需营养素和治疗药物进入中枢神经系统（CNS）的途径。成功地将药物递送至脑肿瘤不仅包括穿透血脑屏障，还包括递送至肿瘤细胞并被肿瘤细胞吸收。大氨基酸转运蛋白 1 (LAT1) 满足所有必要的要求 - 在 BBB 两侧表达、在几乎所有肿瘤细胞（无论来源组织）中高表达、以及在非增殖细胞中极低的内源表达。此外，它的特点是具有相当大的能力，可用于输送有治疗意义的量的化疗剂。 LAT1活性与细胞增殖、肿瘤生长相关，并且与疾病预后和患者存活率呈负相关。 Quadriga BioSciences 将利用 NIH/NCI 资金，将细胞毒性药物的特性与 LAT1 介导的癌细胞递送结合起来，设计新型抗癌药物。拟议的 SBIR 研究第一阶段的目标是确定 LAT1 转运的抗癌药物是否是开发治疗脑肿瘤的化疗药物的可行选择。我们策略的成功被定义为 LAT1 的活性和高度特异性。在目标 1 中，将合成一组化合物，它们是“混合”化合物，其中已知的细胞毒性部分与 LAT1 特异性识别元件进行化学融合。这些化合物将在目标 2 中测试 LAT1 活性和特异性。LAT1 转运将在体外使用测定 HEK 细胞中的 [3H]-加巴喷丁摄取竞争以及直接摄取与 LC/MS/MS 检测相结合。 LAT1 相对于其他氨基酸转运蛋白的特异性将通过表达这些转运蛋白的细胞中的[3H]-苯丙氨酸摄取竞争来测试。将通过体外跨孔转运测定来评估化合物可能的BBB转运。在目标 3 中，评估了化合物对癌细胞系和骨髓祖细胞的细胞毒性（LD50 和 LD100）。为了排除测试化合物通过LAT1以外的方式进入细胞，将确定LAT1抑制剂对细胞毒性的影响。成功的化合物将通过选择性积累对表达 LAT1 的癌细胞系产生细胞毒性，并且对骨髓祖细胞没有细胞毒性。在预计的第二阶段研究中，成功的化合物将在经过验证的各种脑肿瘤异种移植模型中进行体内测试。