Novel ODC Inhibitor Phaseolotoxin in Neuroblastoma

神经母细胞瘤中的新型 ODC 抑制剂菜豆毒素

• 批准号: 9767726
• 负责人: ANDRE S BACHMANN
• 金额: $ 25.95万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-21 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9767726
• 关键词: Antibody Therapy; Apoptosis; Back; Binding; Biological; Biological Assay; Calorimetry; Cell Cycle; Cell Line; Cell Proliferation; Cells; Characteristics; Chemicals; Chemoprevention; Chemopreventive Agent; Child; Clinic; Clinical; Clinical Research; Colon Carcinoma; Computer Assisted; Core Facility; Crystallization; DL-alpha-Difluoromethylornithine; Disease; Disease-Free Survival; Docking; Dose; Drug Targeting; Entropy; Enzyme Inhibition; Enzyme Kinetics; Enzymes; Exhibits; FDA approved; Future; Gene Amplification; Grant; Half-Life; High Dose Chemotherapy; Homologous Gene; In Vitro; Infant; Investigation; Kidney; Kinetics; Letters; MYCN gene; Malignant Childhood Neoplasm; Malignant Neoplasms; Maps; Measures; Modeling; Mus; Natural Products; Neuroblastoma; Oncogenes; Operative Surgical Procedures; Ornithine Decarboxylase; Ornithine Decarboxylase Inhibitor; Patients; Periodicity; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Polyamines; Prevention; Probability; Process; Radiation therapy; Reaction; Relapse; Resistance; Roentgen Rays; Series; Solid; Solubility; Stem cell transplant; Structure; Survival Rate; Sympathetic Nervous System; Testing; Therapeutic; Therapeutic Agents; Time; Tissues; Titrations; Trypanosomiasis; Tumor Tissue; Xenograft Model; analog; base; bench to bedside; cancer cell; cancer type; cell growth; design; efficacy study; efficacy testing; enthalpy; high risk; improved; in vivo; inhibitor/antagonist; neoplastic cell; neuroblastoma cell; novel; novel therapeutics; oncology; pre-clinical; preclinical development; preclinical evaluation; preclinical study; relapse risk; screening; stoichiometry; success; targeted treatment; treatment planning; treatment response; tumor; tumor growth; tumor progression; tumor xenograft

ABSTRACT Neuroblastoma (NB) is an aggressive childhood cancer in which the MYC homologue MYCN acts as an oncogene typically activated to drive tumor progression. NB is a lethal tumor and the survival rate of children with high-risk (stage IV) disease is dismal despite an intense treatment plan that includes a plethora of high- dose chemotherapies, surgery, stem cell transplant, radiation, and antibody therapy. MYCN and ornithine decarboxylase (ODC) are frequently deregulated in NB, and ODC-dependent polyamines control p27/Rb- regulated tumor progression. MYC(N) is a direct activator of ODC and ODC is a validated drug target in NB and other MYC(N)-driven tumors. Difluoromethylornithine (DFMO) is the only ODC inhibitor in the clinic today. DFMO is a FDA-approved anti-protozoan drug (trypanosomiasis) that is effective in the chemoprevention of colon cancer and that has entered clinical studies for children with relapsed NB. Unfortunately, DFMO is required at very high doses due to its short half-life, high solubility, and fast renal elimination/clearance. Therefore, the search for a new ODC inhibitor with higher potency and superior pharmacological profile is warranted. We have previously discovered that the natural product phaseolotoxin (PT) inhibits ODC and exhibits antiproliferative activity in cancer cells. The central hypothesis of this proposal is that novel PT- inspired PSorn-analogs designed for this study are more potent ODC inhibitors than PT or DFMO and, therefore, more effectively inhibit ODC-dependent NB tumor growth in vivo. The objective of this proposal is to identify the most active PSorn-analog with best PK characteristics and confirm its validity as a novel therapeutic agent against NB. In Aim 1 we will chemically synthesize a series of PSorn-analogs for biological & preclinical investigation. We will prepare 17-20 (linear & cyclic) PSorn-analogs. In Aim 2 we will test the hypothesis that these novel PSorn-analogs are potent ODC inhibitors in vitro. In Aim 3 we will test the efficacy of PSorn-analogs in cell-based oncology models and tumor xenograft studies. Active analogs will be selected through a testing funnel that includes: ODC activity and enzyme kinetic assays, NB cell-based oncology models using a panel of 24 well-characterized (MYCN amplified and MYCN non-amplified) NB cell lines with various ODC expression levels, in vitro ADME, and NB tumor xenograft efficacy studies in mice with DFMO- resistant NB cells. Isothermal calorimetry (ITC) and X-ray crystallization will be employed with most active PSorn-analogs to verify ODC binding domains and to aid further SAR-based analog optimization. In summary, these studies will provide the solid groundwork for further preclinical development of an entirely new class of ODC-targeted therapeutic agents with applications in the treatment and prevention of (DFMO-resistant) NB, colon cancer, and other MYC(N)-driven cancer types.

抽象的 神经母细胞瘤 (NB) 是一种侵袭性儿童癌症，其中 MYC 同源物 MYCN 充当 癌基因通常被激活以驱动肿瘤进展。 NB是一种致死性肿瘤，儿童的生存率 尽管采取了包括大量高风险（IV 期）疾病在内的严格治疗计划，但患有高风险（IV 期）疾病的情况仍然令人沮丧。 剂量化疗、手术、干细胞移植、放射和抗体治疗。 MYCN 和鸟氨酸 脱羧酶 (ODC) 在 NB 中经常失调，ODC 依赖性多胺控制 p27/Rb- 调节肿瘤进展。 MYC(N) 是 ODC 的直接激活剂，ODC 是 NB 中经过验证的药物靶点 和其他 MYC(N) 驱动的肿瘤。二氟甲基鸟氨酸 (DFMO) 是目前临床上唯一的 ODC 抑制剂。 DFMO 是 FDA 批准的抗原虫药物（锥虫病），可有效化学预防 结肠癌，并已进入针对复发性 NB 儿童的临床研究。不幸的是，DFMO 由于其半衰期短、溶解度高和肾脏消除/清除速度快，因此需要非常高的剂量。 因此，寻找一种具有更高效力和更优越药理学特征的新型 ODC 抑制剂是当务之急。 保证。我们之前发现天然产物菜豆毒素 (PT) 可以抑制 ODC 和 在癌细胞中表现出抗增殖活性。该提案的中心假设是新颖的 PT- 受启发，为本研究设计的 PSorn 类似物是比 PT 或 DFMO 更有效的 ODC 抑制剂， 因此，更有效地抑制体内ODC依赖性NB肿瘤的生长。该提案的目的是 确定最活跃且具有最佳 PK 特性的 PSorn 类似物，并确认其作为新型药物的有效性 NB治疗剂。在目标 1 中，我们将化学合成一系列 PSorn 类似物，用于生物和 临床前研究。我们将准备 17-20 个（线性和循环）PSorn 类似物。在目标 2 中，我们将测试 假设这些新型 PSorn 类似物在体外是有效的 ODC 抑制剂。在目标3中我们将测试功效 PSorn 类似物在细胞肿瘤模型和肿瘤异种移植研究中的应用。将选择活性类似物 通过测试漏斗，其中包括：ODC 活性和酶动力学测定、NB 基于细胞的肿瘤学 使用一组 24 个经过充分表征（MYCN 扩增和 MYCN 非扩增）的 NB 细胞系进行模型 DFMO-小鼠中的各种 ODC 表达水平、体外 ADME 和 NB 肿瘤异种移植功效研究 耐药NB细胞。大多数活性物质将采用等温量热法 (ITC) 和 X 射线结晶法 PSorn 类似物可验证 ODC 结合域并帮助进一步基于 SAR 的模拟优化。总之， 这些研究将为进一步临床前开发一类全新的药物奠定坚实的基础 ODC 靶向治疗剂可用于治疗和预防（DFMO 耐药）NB、 结肠癌和其他 MYC(N) 驱动的癌症类型。

项目成果

Structural basis of binding and inhibition of ornithine decarboxylase by 1-amino-oxy-3-aminopropane.

1-氨基-氧基-3-氨基丙烷结合和抑制鸟氨酸脱羧酶的结构基础。

• 发表时间: 2021
• 作者: Zhou, X Edward;Suino;Schultz, Chad R;Aleiwi, Bilal;Brunzelle, Joseph S;Lamp, Jared;Vega, Irving E;Ellsworth, Edmund;Bachmann, André S;Melcher, Karsten
• 通讯作者: Melcher, Karsten

Allicin, a Potent New Ornithine Decarboxylase Inhibitor in Neuroblastoma Cells.

大蒜素，神经母细胞瘤细胞中一种有效的新型鸟氨酸脱羧酶抑制剂。

• DOI: 10.1021/acs.jnatprod.0c00613
• 发表时间: 2020-08-28
• 期刊: Journal of natural products
• 影响因子: 5.1
• 作者: Schultz, Chad R.;Gruhlke, Martin C. H.;Slusarenko, Alan J.;Bachmann, Andre S.
• 通讯作者: Bachmann, Andre S.