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: 10474449
• 负责人: Amanda M Schalk
• 金额: $ 83.68万
• 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 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10474449
• 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; 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：设计酶（EBD）正在开发一种更安全的天冬酰胺酶（ASNase），以最大化 这种独特药物的潜在临床应用。 ASNases是全身耗尽的酶药物 从血液中天冬酰胺。在美国，第一线ASNase是Oncaspar，这是大肠杆菌 ASNase（ECA）。 Oncaspar不耐受的患者切换到Naked Erwinia asnase（ERA，Erwinaze）。尽管 作为小儿急性淋巴细胞白血病（全）的关键药物，当前FDA批准的副作用 在成年人中，ASNase是如此明显，以至于他们的使用得到了很大的避免。这些副作用也阻止了使用 急性髓样白血病（AML）和胰腺，卵巢或三阴性乳腺癌中的ASN酶 尽管有强有力的证据表明ASNase将有效地治疗这些癌症。 ECA/ERA STEM的副作用 从i）它们的免疫原性，由于它们的细菌起源和ii）其谷氨酰胺酶（GLNase）共同活性。到 将这种药物的使用扩展到成人所有患者和其他适应症，非常需要ASNase （i）免疫原性降低，（ii）缺乏GLNase共同活性，并结合了（iii）长的体内持久性。 为了减轻免疫原性，EBD正在从豚鼠（GPA）中开发出第一个哺乳动物的心酶（GPA） 与细菌ECA/ERA相比，与人类ASNase的身份更加接近。减少免疫学 更大的风险，我们采用了一种结构引导的策略来人性化GPA并产生GPAHUM。一个添加 Gpahum的优势在于它本质上是无GLN酶的，从而降低了脱靶药物毒性。 EBD 成功地交付了以下STTR 1阶段里程碑：1）11个网站特异性的Gpahum版本 开发以增加其半衰期和最好的变体PEG-GPAHUM，T1/2增加了约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，将我们的分子对抗市场领导者进行基准测试，这是一种非常不可否认的优势毒性 观察到单一和重复给药研究中PEG-GPAHUM与Oncaspar的概况，大大降级风险 开发管道中的PEG-GPAHUM。 我们完成的I-Corps计划，在所有领域中对领导者和利益相关者进行了> 130次访谈。 PEG-GPAHUM的预期可采用性，成为一流的一流。该分子会提供相似的 治疗功效，剂量频率降低和注射药物的总量，预测较少 患者的积累毒性，降低了与药物相关的治疗成本和易用性。这个SBIR阶段 2提案将为PEG-GPAHUM制定适当的GMP扩大制造协议，2） 通过关键的辅助研究和3）使用AML作为患者分层的可行生物标志物，以AML为 扩展指示的第一个示例。未来的SBIR阶段2B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药物具有直接临床意义，对所有人以及将来可能扩展到其他指示 未满足的需求。