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.

项目概述：酶设计（EbD）正在开发一种更安全的L-天冬酰胺酶（ASNase）， 这种独特药物的潜在临床应用。天冬氨酸转氨酶是一种酶药物，系统地消耗L- 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 其他血液恶性肿瘤（如急性髓性白血病）和实体瘤（如骨髓瘤）中的ASNase 胰腺癌、卵巢癌或三阴性乳腺癌），尽管有强有力的证据表明ASNases是有效的 治疗这些癌症。EcA/ErA的副作用源于i）它们的免疫原性，由于它们的细菌毒性， 来源和ii）它们的L-丙氨酸氨基转移酶共活性。将该药物的使用扩展到成人ALL患者和其他 在适应症中，需要具有（i）降低的免疫原性，（ii）缺乏L-丙氨酸氨基转移酶辅酶A的ASNase。 活性，结合（iii）长的体内持久性。为了减轻免疫原性，EbD正在开发 第一种哺乳动物ASNase。与细菌EcA/ErA形成鲜明对比，它们与人类非常遥远。 EbD是一种同源物，EbD正在开发豚鼠ASNase（GpA），其在身份上与人类ASNase更接近。 此外，通过遵循人源化过程，EbD增加了GpA与人GpA的%序列同一性。 同源物总之，这种人源化酶，称为GpAhum，预计免疫原性要低得多 与细菌的天冬氨酸酶相比。这种酶的另一个优点是它本质上不含GLNase， 从而降低脱靶药物毒性。我们有大量的数据表明， GpAhum，毒性小。然而，GpAhum的半衰期（t1/2）对于临床使用不是最佳的。而PEG化 提供了一种增加t1/2的解决方案，如OnCaspar所示，最近的临床数据报告了抗PEG抗体 在以前用OnCaspar治疗的患者中，明确表明PEG化不是临床治疗的可行途径。 新的ASNase因此，为了实现t1/2增加，我们转向人血清白蛋白（HSA）结合 通过将我们的ASNase融合到免疫中性的人血清白蛋白结合结构域（ABD）的技术。 该产品ABD-GpAhum的血液循环时间比亲本GpAhum的血液循环时间增加了一倍多， 结合亲和力在小鼠异种移植模型中的体内疗效证实了其上级效力：1剂ABD-GpAhum 相当于3次剂量的GpAhum，允许相似的治疗效果，降低给药频率和总剂量 注射的药物量。这预示着患者的累积毒性较小，药物相关治疗费用降低 以及医院相关人员的易用性。该提案将进一步评估ABD-GpAhum的抗ALL疗效 并提供概念验证毒性数据，以确认ABD-GpAhum确实是一种可行的，安全的和有竞争力的药物。 治疗候选药物，从而为通过IND使能研究推进其提供了依据。