A novel glutaminase-free mammalian asparaginase with minimized immunogenicity to enable expanded use in cancer therapy

一种新型无谷氨酰胺酶的哺乳动物天冬酰胺酶，具有最小化的免疫原性，可扩大在癌症治疗中的应用

• 批准号: 10262960
• 负责人: Amanda M Schalk
• 金额: --
• 依托单位: ENZYME BY DESIGN, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10262960
• 关键词: Acute; Acute Lymphocytic Leukemia; Acute Myelocytic Leukemia; Adopted; Adoption; Adult; Adult Acute Lymphocytic Leukemia; Asparagine; Benchmarking; Biological; Biological Markers; Biological Process; Blood; Blood Circulation; Breast; Canis familiaris; Cardiopulmonary; Cavia; Cell Line; Childhood Acute Lymphocytic Leukemia; Chimera organism; Clinical; Clinical Trials; Cytarabine; Cytogenetics; Data; Development; Dose; Doxorubicin; Drug Kinetics; Drug Targeting; Drug toxicity; Drug usage; Endotoxins; Engineering; Enzymes; Erwinia; Escherichia coli; FDA approved; Formulation; Frequencies; Future; Glutaminase; Half-Life; Human; Immunologics; In Vitro; Innovation Corps; Interview; Laboratories; Lead; Malignant Neoplasms; Methods; Monosomy 7; Mus; Myelogenous; Nebraska; Outcome; Ovarian; Pancreas; Patients; Pegaspargase; Pharmaceutical Preparations; Pharmacology; Pharmacology and Toxicology; Phase; Preparation; Probability; Process; Production; Property; Protocols documentation; Quality Control; Rattus; Reporting; Rights; Risk; Rivers; Site; Small Business Innovation Research Grant; Small Business Technology Transfer Research; Structure; Testing; Therapeutic; Time; Toxic effect; Toxicology; Treatment Efficacy; Universities; Variant; Xenograft Model; acute myeloid leukemia cell; arm; asparaginase; cancer therapy; cell bank; clinical application; clinical efficacy; clinical material; clinical practice; clinically relevant; commercialization; cost; design; first-in-human; immunogenic; immunogenicity; in vivo; manufacturing process; manufacturing scale-up; novel; patient biomarkers; patient derived xenograft model; patient stratification; prevent; programs; response; scale up; side effect; standard of care; stem; triple-negative invasive breast carcinoma

Project Summary: Enzyme by Design (EbD) is developing a safer asparaginase (ASNase) to maximize the potential clinical applications of this unique drug. ASNases are enzyme drugs that systemically deplete asparagine from the blood. In the US, the 1st-line ASNase is Oncaspar, a PEGylated version of the E. coli ASNase (EcA). Patients intolerant of Oncaspar switch to the naked Erwinia ASNase (ErA, Erwinaze). Despite being key drugs in pediatric acute lymphoblastic leukemia (ALL), the side effects of current FDA-approved ASNases are so pronounced in adults that their use is largely avoided. These side effects also prevent the use of ASNases in acute myeloid leukemia (AML) and in pancreatic, ovarian or triple-negative breast cancers, despite strong evidence that ASNases would be effective in treating those cancers. Side effects of EcA/ErA stem from i) their immunogenicity, due to their bacterial origin and ii) their glutaminase (GLNase) co-activity. To expand the use of this drug to adult ALL patients and to other indications, there is a strong need for an ASNase with (i) reduced immunogenicity, (ii) lack of GLNase co-activity, combined with (iii) long in vivo persistence. To mitigate the immunogenicity, EbD is developing the first mammalian ASNase from the guinea pig (GpA) that is much closer in identity to the human ASNase compared to the bacterial EcA/ErA. To reduce the immunological risk even more, we employed a structure-guided strategy to humanize GpA, generating GpAhum. An added advantage of GpAhum is that it is intrinsically GLNase-free, thereby reducing off-target drug toxicity. EbD successfully delivered the following STTR Phase 1 milestones: 1) 11 site-specific PEGylated versions of GpAhum were developed to increase its half-life and the best variant, PEG-GpAhum with ~3-fold increase in t1/2 was identified. 2) in T-ALL mouse xenograft models, PEG-GpAhum q.wk was more potent than GpAhum t.i.w, and 3) most importantly, benchmarking our molecule against the market leader, a strikingly undeniable superior toxicity profile of PEG-GpAhum vs. Oncaspar in both single and repeat dosing studies was observed, greatly de-risking PEG-GpAhum in the development pipeline. Our completed I-Corps programs with >130 interviews with leaders and stakeholders in the ALL field validated the expected adoptability of PEG-GpAhum, becoming best-in-class. This molecule would deliver similar therapeutic efficacy with reduced dosing frequency and total amount of drug injected, predicting less accumulated toxicity in patients, lowered drug-related therapy cost and increased ease of use. This SBIR Phase 2 proposal will 1) develop appropriate GMP scale-up manufacturing protocols for PEG-GpAhum, 2) advance it through key IND-enabling studies and 3) confirm a viable biomarker for patient stratification, using AML as the first example of expanded indication. Future SBIR Phase 2b will support the completion of the IND application package and GMP material for clinical use. Successful development of PEG-GpAhum will supply a much safer ASNase drug with immediate clinical implications for ALL and likely future expansion to additional indications of unmet need.

项目摘要：Enzyme by Design (EbD) 正在开发一种更安全的天冬酰胺酶 (ASNase)，以最大限度地提高 这种独特药物的潜在临床应用。 ASNases 是一种酶药物，可以系统性地消耗 从血液中提取天冬酰胺。在美国，第一线 ASNase 是 Oncaspar，它是大肠杆菌的 PEG 化版本 ASNase (EcA)。不耐受 Oncaspar 的患者改用裸欧文氏菌 ASNase（ErA、Erwinaze）。尽管 作为小儿急性淋巴细胞白血病 (ALL) 的关键药物，目前 FDA 批准的副作用 ASNases 在成人中如此明显，以至于人们基本上避免使用它们。这些副作用也阻碍了使用 急性髓系白血病 (AML) 和胰腺癌、卵巢癌或三阴性乳腺癌中的 ASNases， 尽管有强有力的证据表明 ASNASes 可有效治疗这些癌症。 EcA/ErA 干的副作用 i) 由于细菌来源而具有免疫原性，以及 ii) 谷氨酰胺酶 (GLNase) 协同活性。到 将这种药物的使用扩大到成人 ALL 患者和其他适应症，强烈需要 ASNase 具有 (i) 免疫原性降低，(ii) 缺乏 GLNase 共活性，以及​​ (iii) 体内持久性长。 为了减轻免疫原性，EbD 正在开发第一种来自豚鼠的哺乳动物 ASNase (GpA)， 与细菌 EcA/ErA 相比，它在同一性上更接近人类 ASNase。为了降低免疫 风险更大，我们采用了结构引导策略来人性化 GpA，生成 GpAhum。添加了一个 GpAhum 的优势在于它本质上不含 GLNase，从而减少了脱靶药物毒性。乙二胺 成功实现了以下 STTR 第 1 阶段里程碑：1) 11 个特定位点的 PEG 化版本的 GpAhum 开发是为了延长其半衰期，最佳变体 PEG-GpAhum 的 t1/2 增加约 3 倍 确定。 2) 在 T-ALL 小鼠异种移植模型中，PEG-GpAhum q.wk 比 GpAhum t.i.w 更有效，3) 最重要的是，将我们的分子与市场领导者进行基准比较，其具有不可否认的卓越毒性 在单次和重复给药研究中观察到 PEG-GpAhum 与 Oncaspar 的比较，大大降低了风险 PEG-GpAhum 正在开发中。 我们已完成 I-Corps 计划，对 ALL 领域的领导者和利益相关者进行了超过 130 次采访，并经过验证 PEG-GpAhum 的预期采用率，成为同类最佳。该分子将提供类似的 通过减少给药频率和注射药物总量来提高治疗效果，预测更少 减少患者体内累积的毒性，降低药物相关的治疗成本并提高使用的便利性。此 SBIR 阶段 2 提案将 1) 为 PEG-GpAhum 制定适当的 GMP 放大生产方案，2) 推进它 通过关键的 IND 支持研究，以及 3) 确认用于患者分层的可行生物标志物，使用 AML 作为 扩展指示的第一个示例。未来的 SBIR 2b 阶段将支持 IND 申请的完成 临床使用的包装和 GMP 材料。 PEG-GpAhum的成功开发将提供更安全的 ASNase 药物对 ALL 具有直接的临床意义，并且未来可能扩展到其他适应症 未满足的需求。