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.

项目概述：酶设计（EbD）正在开发一种更安全的天冬酰胺酶（ASNase），以最大限度地提高酶的活性。 这种独特药物的潜在临床应用。天冬氨酸酶是酶类药物， 血液中的天冬酰胺在美国，一线ASNase是OnCaspar，一种E.杆菌 ASNase（EcA）.对OnCaspar不耐受的患者切换到裸欧文氏菌ASNase（ErA，Erwinaze）。尽管 作为儿科急性淋巴细胞白血病（ALL）的关键药物，目前FDA批准的 ASNase在成年人中非常明显，因此在很大程度上避免使用。这些副作用也阻止了使用 急性髓性白血病（AML）和胰腺癌、卵巢癌或三阴性乳腺癌中的ASNase， 尽管有强有力的证据表明ASNases在治疗这些癌症中是有效的。EcA/ErA股骨柄的副作用 来自i）它们的免疫原性，由于它们的细菌来源，和ii）它们的谷氨酰胺酶（GLNase）共活性。到 为了将该药物的使用范围扩大到成人ALL患者和其他适应症，对ASNase的需求非常大 具有（i）降低的免疫原性，（ii）缺乏GLNA酶共活性，以及（iii）长的体内持久性。 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，其身份更接近于人ASNase。为了减少免疫 风险更大，我们采用了结构指导的策略来人源化GpA，产生GpAhum。一个额外 GpAhum的优势在于其本质上不含GLNase，从而降低脱靶药物毒性。EBD 成功交付了以下STTR 1期里程碑：1）11个位点特异性聚乙二醇化版本的GpAhum 为了增加其半衰期，开发了最佳变体PEG-GpAhum，其t1/2增加约3倍， 鉴定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，以及3） 最重要的是，将我们的分子与市场领先者进行基准测试， 在单次和重复给药研究中均观察到PEG-GpAhum与OnCaspar的特征， PEG-GpAhum在开发管道中。 我们已完成的I-Corps计划，与ALL领域的领导者和利益相关者进行了超过130次访谈， PEG-GpAhum的预期适应性，成为一流的。这种分子可以提供类似于 降低给药频率和注射药物总量的治疗效果，预测 在患者中累积毒性，降低药物相关治疗成本并增加易用性。此SBIR阶段 2提案将1）为PEG-GpAhum制定适当的GMP规模扩大生产方案，2）推进 通过关键的IND使能研究和3）确认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SBIR第2b阶段将支持IND申请的完成 包装和GMP材料，供临床使用。PEG-GpAhum的成功开发将提供更安全的 ASNase药物对ALL具有直接临床意义，未来可能扩展到其他适应症 未满足的需求