Extending the in vivo half-life of a novel engineered mammalian asparaginase via its binding to human serum albumin

通过与人血清白蛋白的结合延长新型工程哺乳动物天冬酰胺酶的体内半衰期

• 批准号: 10075744
• 负责人: Amanda M Schalk
• 金额: $ 40万
• 依托单位: ENZYME BY DESIGN, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10075744
• 关键词: Acute; Acute Lymphocytic Leukemia; Acute Myelocytic Leukemia; Adoption; Adult; Adult Acute Lymphocytic Leukemia; Affinity; Albumins; Amino Acids; Amylases; Antibodies; Asparagine; Bilirubin; Binding; Biological; Blood; Blood Chemical Analysis; Blood Circulation; Blood Circulation Time; Body Weight Changes; Bone Marrow; Breast; Cavia; Cell Line; Cells; Childhood Acute Lymphocytic Leukemia; Chimera organism; Clinic; Clinical; Clinical Data; Clinical Trials; Data; Data Reporting; Distant; Dose; Drug Kinetics; Drug Targeting; Drug toxicity; Drug usage; Engineering; Enzymes; Erwinia; Escherichia coli; Eye; FDA approved; Fatty acid glycerol esters; Feedback; Formulation; Frequencies; Glutaminase; Half-Life; Hematologic Neoplasms; Histology; Histopathology; Homologous Gene; Hospitals; Human; Immunologics; Laboratories; Lead; Liver; Malignant Neoplasms; Measurable; Modeling; Mus; Myelogenous; Ovarian; PTPRC gene; Pancreas; Patients; Pegaspargase; Pharmaceutical Preparations; Phase; Plasma Proteins; Posture; Process; Production; Property; Recurrence; Renal function; Rivers; Safety; Serum Albumin; Signal Transduction; Solid Neoplasm; Spleen; Stains; Technology; The Jackson Laboratory; Time; TimeLine; Toxic effect; Toxicology; Treatment Efficacy; Urea Nitrogen; Vacuole; Variant; Xenograft Model; Xenograft procedure; asparaginase; base; bioluminescence imaging; clinical application; clinical efficacy; clinical practice; clinically relevant; commercialization; cost; design; efficacy study; head-to-head comparison; immunogenic; immunogenicity; in vivo; novel; pediatric patients; peripheral blood; phase 1 study; prevent; side effect; standard of care; stem; therapeutic candidate; triple-negative invasive breast carcinoma

PROJECT SUMMARY: Enzyme by Design (EbD) is developing a safer L-asparaginase (ASNase) to maximize the potential clinical applications of this unique drug. ASNases are enzyme drugs that systematically deplete L- 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 other hematological malignancies (e.g. acute myeloid leukemia) and in solid tumors (e.g. 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 L-glutaminase co-activity. To expand the use of this drug to adult ALL patients and to other indications, there is a need for an ASNase with (i) reduced immunogenicity, (ii) lack of L-glutaminase co- activity, combined with (iii) long in vivo persistence. To mitigate the immunogenicity, EbD is developing the first mammalian ASNase. In sharp contrast to the bacterial EcA/ErA, which are very distant from the human homolog, EbD is developing the guinea pig ASNase (GpA), which is much closer in identity to the human ASNase. Moreover, by following a humanization process, EbD increased GpA’s % sequence identity to the human homolog. Together, this humanized enzyme, referred to as GpAhum, is predicted to be much less immunogenic compared to the bacterial ASNases. An added advantage of this enzyme is that it is intrinsically GLNase-free, thereby reducing off-target drug toxicity. We have extensive data that shows strong in vivo anti-ALL potency of GpAhum with little toxicity. However, the half-life (t1/2) of GpAhum is not optimal for clinical use. Whereas PEGylation offers one solution for increasing the t1/2, as seen in Oncaspar, recent clinical data reporting anti-PEG antibodies in patients previously treated with Oncaspar made it clear that PEGylation is not a viable path to the clinic for any new ASNase. Therefore, to achieve an increased t1/2, we pivoted to human serum albumin (HSA)-binding technology via fusing our ASNase to an immunologically neutral human serum albumin binding domain (ABD). The product, ABD-GpAhum, more than doubled the parental GpAhum’s blood circulation time with excellent HSA binding affinity. In vivo efficacy in mouse xenograft models confirms its superior potency: 1 dose of ABD-GpAhum is equivalent to 3 doses of GpAhum, allowing similar therapeutic efficacy with reduced dosing frequency and total amount of drug injected. This predicts less accumulated toxicity in patients, lowered drug-related therapy cost and ease of use for hospital-related staffs. This proposal will further evaluate the anti-ALL efficacy of ABD-GpAhum and supply proof-of-concept toxicity data to confirm that ABD-GpAhum is indeed a viable, safe and competitive therapeutic candidate, thus providing the rationale for advancing it through IND-enabling studies.

项目摘要：Enzyme by Design (EbD) 正在开发一种更安全的 L-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。 ASNase 是一种酶药物，可以系统性地消耗 L- 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 ASNase 在其他血液恶性肿瘤（例如急性髓系白血病）和实体瘤（例如骨髓瘤）中的作用 胰腺癌、卵巢癌或三阴性乳腺癌），尽管有强有力的证据表明 ASNase 有效 治疗这些癌症。 EcA/ErA 的副作用源于 i) 它们的免疫原性，由于其细菌 起源和 ii) 它们的 L-谷氨酰胺酶共同活性。将这种药物的使用范围扩大到成人 ALL 患者和其他患者 适应症，需要一种具有以下特征的 ASNase：(i) 免疫原性降低，(ii) 缺乏 L-谷氨酰胺酶辅酶 活性，与（iii）体内持久性相结合。为了减轻免疫原性，EbD 正在开发 第一个哺乳动物 ASNase。与距离人类很远的细菌EcA/ErA形成鲜明对比 同源物，EbD 正在开发豚鼠 ASNase (GpA)，它与人类 ASNase 的特性更加接近。 此外，通过遵循人源化过程，EbD 增加了 GpA 与人类的序列同一性百分比 同系物。总之，这种被称为 GpAhum 的人源化酶预计免疫原性要低得多 与细菌 ASNase 相比。这种酶的另一个优点是它本质上不含 GLNase， 从而减少药物的脱靶毒性。我们有大量的数据显示出强大的体内抗 ALL 效力 GpAhum毒性很小。然而，GpAhum 的半衰期 (t1/2) 并不适合临床使用。而聚乙二醇化 提供了一种增加 t1/2 的解决方案，如 Oncaspar 中所示，最近报告抗 PEG 抗体的临床数据 先前接受过 Oncaspar 治疗的患者的研究表明，聚乙二醇化并不是进入临床的可行途径 任何新的 ASNase。因此，为了提高 t1/2，我们转向人血清白蛋白 (HSA) 结合 通过将我们的 ASNase 与免疫中性人血清白蛋白结合域 (ABD) 融合的技术。 该产品ABD-GpAhum，具有优异的HSA，使亲本GpAhum的血液循环时间延长了一倍以上 结合亲和力。小鼠异种移植模型的体内功效证实了其卓越功效：1 剂 ABD-GpAhum 相当于 3 剂 GpAhum，可实现相似的治疗效果，同时减少给药频率和总剂量 注射的药物量。这预示着患者的累积毒性会减少，药物相关的治疗成本也会降低 以及医院相关工作人员的易用性。该提案将进一步评估 ABD-GpAhum 的抗 ALL 功效 并提供概念验证毒性数据，以确认 ABD-GpAhum 确实是一种可行、安全且具有竞争力的药物 治疗候选者，从而为通过 IND 支持研究推进它提供了理由。