title: GMP production and characterization of leucine zipper-tagged tumor necrosis factor-related apoptosis-inducing ligand (LZ-TRAIL) for phase I clinical trial.
abstract: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) exhibits potent antitumor activity in a wide range of cancers without deleterious side effects on normal tissues. Several TRAIL derivatives have been developed to improve its pharmacokinetics and therapeutic effects through strategies such as adding a leucine zipper to increase the circulation half-life. To obtain clinical grade LZ-TRAIL for phase I clinical trial, a single batch of 30 L bioreactor culture was performed using the Escherichia coli BL21 (DE3) strain expressing the recombinant LZ-TRAIL. A robust LZ-TRAIL production fermentation process was developed, which could be scaled up from 5L to 50 L, and had a titer of approximately 1.4 g/l. A four-step purification strategy was carried out to obtain a final product with over 95% purity and 45% yield. The final material was filter sterilized, aseptically vialed, and stored at 4°C, and comprehensively characterized using multiple assays (vialed product was sterile, purity was 95%, aggregates were <5%, potency revealed IC50 of 9 nM on MDA-MB-231 cells, and the endotoxin level was <0.25 U/mg). The purity, composition, and functional activities of the molecule were confirmed. in vivo investigations indicated that LZ-TRAIL has better antitumor potency in three Xenograft tumor models compared to TRAIL (95-281). LZ-TRAIL also showed improved pharmacokinetic and safety profiles in cynomolgus monkeys without abnormalities associated with drug exposure. In conclusion, the scalable synthesis of LZ-TRAIL is useful for production of phase I clinical trial material. These preclinical investigations warrant further clinical development of this product for cancer therapy.
authors.affiliation: Department of Pharmacology, Binzhou Medical University, Shandong Province, Yantai 256603, China. Electronic address: jing_jiang1974@sina.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Shandong Simcere-Medgenn Biopharmaceutical Co., Ltd., Shandong Province, Yantai 264006, China.
authors.forename: Xiaobin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Shandong Simcere-Medgenn Biopharmaceutical Co., Ltd., Shandong Province, Yantai 264006, China.
authors.forename: Leixiu
authors.initials: L
authors.lastname: Deng
authors.affiliation: Shandong Simcere-Medgenn Biopharmaceutical Co., Ltd., Shandong Province, Yantai 264006, China.
authors.forename: Peipei
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Shandong Simcere-Medgenn Biopharmaceutical Co., Ltd., Shandong Province, Yantai 264006, China.
authors.forename: Guangjun
authors.initials: G
authors.lastname: Wang
authors.affiliation: Shandong Simcere-Medgenn Biopharmaceutical Co., Ltd., Shandong Province, Yantai 264006, China.
authors.forename: Shifu
authors.initials: S
authors.lastname: Wang
authors.affiliation: Shandong Simcere-Medgenn Biopharmaceutical Co., Ltd., Shandong Province, Yantai 264006, China.
authors.forename: Honghao
authors.initials: H
authors.lastname: Liu
authors.affiliation: Simcere Pharmaceutical R&D Department; Jiangsu Simcere-Medgenn Biopharmaceutical Co., Ltd., Jiangsu Province, Nanjing 210000, China. Electronic address: suyunpeng@simcere.com.
authors.forename: Yunpeng
authors.initials: Y
authors.lastname: Su
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D017321:Clinical Trials, Phase I as Topic; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D022781:Hepatocytes / Q000187:drug effects; D006801:Humans; D008252:Macaca fascicularis; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009369:Neoplasms / Q000188:drug therapy / Q000473:pathology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use; D047368:Tumor Burden / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; C448905:LZ-TRAIL protein, recombinant; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand; D014409:Tumor Necrosis Factor-alpha
keywords: Anticancer; Escherichia coli; Leucine zipper; Preclinical; Safety; TRAIL