Pharmacological and toxicological testing of a novel L-asparaginase

新型L-天冬酰胺酶的药理和毒理测试

• 批准号: 10265351
• 负责人: ARNON LAVIE
• 金额: --
• 依托单位: JESSE BROWN VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10265351
• 关键词: Acute Lymphocytic Leukemia; Acute Myelocytic Leukemia; Address; Adult; Adult Acute Lymphocytic Leukemia; Affect; Amino Acids; Antineoplastic Agents; Apoptosis; Asparagine; Biological; Blood; Blood Circulation; C-terminal; Cancer Patient; Canis familiaris; Catalytic Domain; Cavia; Cells; Childhood Acute Lymphocytic Leukemia; Clinic; Clinical; Clinical Data; Crystallization; Data; Disease; Drug Kinetics; Drug usage; Enzymes; Escherichia coli; Exhibits; FDA approved; Glutaminase; Glutamine; Goals; Hematologic Neoplasms; Homologous Gene; Human; Hydrolysis; Immune system; Immunologics; Investigational Drugs; Investigational New Drug Application; Kinetics; Lead; Legal patent; Leukocytes; Life; Malignant Neoplasms; Malignant neoplasm of pancreas; Mus; Pancreas; Patients; Pectobacterium chrysanthemi; Pegaspargase; Pharmaceutical Preparations; Pharmacology and Toxicology; Property; Protocols documentation; Publishing; Rattus; Reaction; Reporting; Risk; Safety; Side; Site; Solid; Solid Neoplasm; Source; Structure; Testing; Therapeutic; Time; Toxic effect; Variant; Work; acute lymphoblastic leukemia cell; anti-cancer; asparaginase; base; cell killing; clinical efficacy; drug development; drug efficacy; experimental study; humanized mouse; immunogenic; immunogenicity; improved; in vivo; leukemia treatment; mortality; mouse model; neoplastic cell; novel; novel therapeutics; pancreatic cancer patients; patient population; pre-clinical; preclinical efficacy; preclinical study; preclinical toxicity; prevent; reconstitution; side effect; standard of care; success

Project Summary: The goal of this proposal is to perform IND-enabling studies of a significantly safer variant of the anti-cancer biologic drug L-asparaginase. L-asparaginases are enzyme drugs that act to deplete the amino acid asparagine from the blood. Due to toxicity, which is especially pronounced in adults, L- asparaginase treatment is limited to acute lymphoblastic leukemia (ALL), a cancer of white blood cells. One source of toxicity of L-asparaginases is due to their bacterial origin (either from E. coli (Elspar) or Erwinia chrysanthemi (Erwinaze)), making the naked drugs highly immunogenic. The current standard of care is a PEGylated version of Elspar called Oncaspar. While PEGylation reduces, but does not eliminate the immunological challenge of using these drugs, the other toxicity-causing factor remains - this being their L- glutaminase coactivity. Therefore, Oncaspar is limited to ALL, where even its use to treat adult ALL patients is highly limited. Of note, L-asparaginase-associated side effects prevent the use of this unique cancer drug in other hematological malignancies (e.g. acute myeloid leukemia) and in solid tumors (e.g. pancreatic cancer), despite strong evidence that L-asparaginases would be effective in treating those cancers. Hence, there is a clear unmet need for an L-asparaginase with reduced immunogenicity and that lacks L-glutaminase coactivity. Recently, we characterized a guinea pig L-asparaginase (GpA) that possesses the required low KM property for clinical efficacy and that exhibits in vivo tumor cell-killing. Notably, we also discovered that GpA is devoid of the toxicity-causing L-glutaminase co-activity. With ~70% sequence identity to human L- asparaginase, GpA should be less immunogenic compared to the bacterial enzymes that share only ~25% sequence identity with the human enzyme. We recently identified the lead biologic GpA369 which is a stable and active C-terminal truncation of GpA comprising the catalytic domain. Here we will perform the required studies required to bring this novel enzyme drug to patients. In Aim 1 we will increase its sequence identity to the human homolog using a structure-guided approach and identify the optimal PEGylation strategy. Aim 2 will determine the pharmacokinetic properties of the top 3 optimized leads from Aim 1, as well as confirm their anti- cancer efficacy in a human ALL mouse model. In Aim 3, the top variant (optimal combination of PK and anti- cancer efficacy) will proceed to toxicity studies, first in mice, followed by more extensive studies in rats and dogs. Finally, Aim 4 will evaluate the immunogenicity of our enzyme drug in a novel mouse model that has a reconstituted human immune system, and compare our drug to Oncaspar. Together, these studies will bring us to the cusp of submitting an IND application for testing this novel L-asparaginase in patients. Impact is predicted to extend beyond ALL, since the improved safety profile of our L-asparaginase variant would enable its use in multiple cancers for which effective options are sorely lacking and which strong data suggests that an L-asparaginase drug would be effective but is not used due to the unacceptable toxicity profile of current L- asparaginase options.

项目摘要：该提案的目标是对一种明显更安全的变体进行 IND 研究 抗癌生物药物L-天冬酰胺酶的研究。 L-天冬酰胺酶是一种酶药物，可消耗 来自血液的氨基酸天冬酰胺。由于毒性，L-在成人中尤其明显 天冬酰胺酶治疗仅限于急性淋巴细胞白血病 (ALL)，这是一种白细胞癌症。一 L-天冬酰胺酶的毒性来源是其细菌来源（来自大肠杆菌 (Elspar) 或欧文氏菌） 菊花（Erwinaze）），使得裸露药物具有高度免疫原性。目前的护理标准是 Elspar 的聚乙二醇化版本称为 Oncaspar。虽然聚乙二醇化减少但并没有消除 使用这些药物的免疫挑战，另一个引起毒性的因素仍然存在 - 这就是它们的 L- 谷氨酰胺酶协同活性。因此，Oncaspar 仅限于 ALL，甚至用于治疗成人 ALL 患者也很有限。 非常有限。值得注意的是，L-天冬酰胺酶相关的副作用阻止了这种独特的癌症药物在 其他血液恶性肿瘤（例如急性髓系白血病）和实体瘤（例如胰腺癌）， 尽管有强有力的证据表明 L-天冬酰胺酶可有效治疗这些癌症。因此，有一个 对免疫原性降低且缺乏 L-谷氨酰胺酶的 L-天冬酰胺酶的需求明显未得到满足 协同性。最近，我们鉴定了豚鼠 L-天冬酰胺酶 (GpA)，它具有所需的低 KM 具有临床疗效和体内肿瘤细胞杀伤作用。值得注意的是，我们还发现 GpA 缺乏引起毒性的L-谷氨酰胺酶共活性。与人类 L- 具有约 70% 的序列同一性 与仅占约 25% 的细菌酶相比，天冬酰胺酶、GpA 的免疫原性应较低 与人类酶的序列同一性。我们最近鉴定了领先的生物制剂 GpA369，它是一种稳定的 以及包含催化结构域的 GpA 的活性 C 端截短。在这里我们将执行所需的操作 将这种新型酶药物带给患者所需的研究。在目标 1 中，我们将增加其序列同一性至 使用结构引导的方法分析人类同源物并确定最佳的聚乙二醇化策略。目标2将 确定目标 1 中最优化的 3 个先导化合物的药代动力学特性，并确认它们的抗 人类 ALL 小鼠模型中的癌症功效。在目标 3 中，顶级变体（PK 和抗病毒的最佳组合） 癌症功效）将进行毒性研究，首先在小鼠中进行，然后在大鼠和小鼠中进行更广泛的研究 狗。最后，Aim 4 将在一种新型小鼠模型中评估我们的酶药物的免疫原性，该模型具有 重建人体免疫系统，并将我们的药物与 Oncaspar 进行比较。这些研究共同将为我们带来 即将提交 IND 申请以在患者中测试这种新型 L-天冬酰胺酶。影响是 预计将超越 ALL，因为我们的 L-天冬酰胺酶变体的安全性得到改善，将能够 它在多种癌症中的应用，这些癌症严重缺乏有效的选择，并且强有力的数据表明， L-天冬酰胺酶药物可能有效，但由于目前 L-天冬酰胺酶的毒性不可接受而未使用 天冬酰胺酶选项。