Nonclinical safety evaluation and pharmacokinetics of Ab612, a novel anti-L1CAM (CD171) therapeutic antibody candidate against solid cancer

Alpha-synuclein is a 15 kDa protein associated with neurodegenerative diseases such as Parkinson disease and multiple-system atrophy where pathological forms of alpha-synuclein aggregate and become neurotoxic. Here we describe the nonclinical program to support a first-in-human (FIH) single ascending dose (SAD) study for Lu AF82422, a human recombinant, anti-alpha-synuclein monoclonal antibody (mAb) in development for treatment of synucleinopathies. Alpha-synuclein is primarily expressed in brain, peripheral nerves and in blood cells. A tissue cross-reactivity assessment showed that Lu AF82422 binding was generally restricted to nervous tissues. Flow cytometry analysis did not show extracellular surface binding of Lu AF82422 to human platelets, erythrocytes, granulocytes, or lymphocytes, but to a low fraction of monocytes, without any functional consequences on activation or phagocytic capacity. A single dose pharmacokinetic (PK) study in cynomolgus monkeys with dose levels of 1–30 mg/kg confirmed PK properties in the expected range for a mAb with a soluble target, and target engagement was shown as a decrease in free alpha-synuclein in plasma. Four-week repeat-dose toxicity studies were conducted in rats and cynomolgus monkeys at doses up to 600 mg/kg administered intravenously every 10 days. Results showed no treatment-related adverse findings and the no-observed-adverse-effect-level was the highest dose tested. Target engagement was shown in plasma and cerebrospinal fluid. Taken together, the nonclinical data indicated no safety signal of concern and provided adequate safety margins between observed safe doses in animals and the planned dose levels in the FIH SAD study.

Interleukin-13 receptor alpha 2 (IL-13Rα2) is overexpressed in various types of cancers including glioma and melanoma and is known to be associated with increased tumor invasiveness and metastases. Here, we present the preclinical efficacy and toxicity results for a novel, anti-IL-13Rα2 ADC-LM-306. In vitro binding activity and internalization of LM-306 and LM-106 (unconjugated antibody of LM-306) were assessed in human IL-13Rα2 overexpressing and endogenous cell lines using flow cytometry. The antibody-dependent cell-mediated cytotoxicity (ADCC) and direct cytotoxicity were assessed using viability assays. Cross-species reactivity of drug was assessed in cells expressing cynomolgus, rat and mouse IL-13Rα2. In vivo A375 CDX model was used to assess the effect of LM-306 anti-tumor activity. The toxicity of LM-306 was evaluated in repeated-dose intravenous dose range-finding (DRF) toxicity and single intravenous maximum tolerated dose (MTD) study in cynomolgus monkeys. Both LM-106 and LM-306 showed concentration dependent binding affinity to IL-13Rα2 in various cell lines (Table 1). In addition, LM-106 also showed a dose dependent internalization and ADCC and LM-306 showed significant cytotoxicity in all cell lines (Table 1). Furthermore, LM-306 showed comparable binding affinity to human IL-13Rα2 to its unconjugated counterpart LM-106 (KD:7.43E-10 vs 1.77E-9). LM-306 was also found to be cross-reactive toward cynomolgus IL-13Rα2. In vivo, LM-306 showed tumor growth inhibition at a dose of 1, 3, 10 mg/kg given intravenously once weekly, indicating its strong anti-tumor activity. The highest non-severely toxic dose of LM-306 was determined to be 9 mg/kg in the DRF study and MTD was determined to be 18mg/kg in MTD study for male monkey. In summary, LM-306 showed high binding affinity to human IL13Rα2 expressing cells with potent cytotoxicity in vitro and significant in vivo inhibition of tumor growth, suggesting its therapeutic benefit in immuno-oncology. Summary of parameters of LM-106 and LM-306 binding affinity, internalization, ADCC and cytotoxicity Assay Antibody EC50 (nM) HEK293/H_IL-13Rα2 U251 A375 SKMEL5 Binding affinity LM-106 0.99 0.21 0.57 0.1 LM-306 1.47 0.33 0.8 0.12 Internalization LM-106 2.22 - 0.43 - ADCC LM-106 0.16 - 0.03 - LM-306 0.31 - 0.03 - Cytotoxicity (IC50) LM-306 0.03 0.05 0.01 0.005 Citation Format: Wentao Huang, Zhifang Liu, Yifan Li, Heng Pan, Yuan Li, Xia Qin, Da Fei, Runsheng Li. Pre-clinical evaluation of a novel antibody drug conjugate (ADC) LM-306 targeting IL-13Rα2 in immuno-oncology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1881.

Lbl-043, a Novel LILRB4xCD3 T Cell Engager, for the Treatment of Relapsed/Refractory Multiple Myeloma

Background: Multiple myeloma (MM) is a clonal plasma cell proliferative disorder characterized by abnormal increase of monoclonal immunoglobulins. Despite newly developed novel therapeutic agents have greatly improved the outcome of MM patients in the past decades, MM is still an incurable hematological malignancy. Several CD3 bispecific immunotherapies that harness T cells against MM are at various stages of preclinical and clinical development, but most of them are hampered by the refractory and resistant mechanisms. Leukocyte immunoglobulin-like receptor B4 (LILRB4) is an immune checkpoint inhibitory receptor, which is highly expressed on monocytic blasts of AML and myeloid-derived suppressive cells. Most recently it is discovered also highly expressed in MM, in particular rrMM(relapsed/refractory MM). It does not express on hematopoietic stem cells and progenitor cells, therefore make it a promising target for hematological malignancies. Herein a novel T cell engager targeting LILRB4, LBL-043, is developed to specifically kill LILRB4 expressing MM cells. Methods: LBL-043 is a bispecific antibody targeting LILRB4 and CD3, designed using proprietary LeadsBodyTM platform, which is a 2:1 format with two VHH arms targeting LILRB4 with high affinity and one scFv arm targeting CD3 with fine-tuned affinity. The binding affinity of LBL-043 was determined with Fortebio and flow cytometry, and the function activity was measured using reporter gene and TDCC assays on MM cell lines. The anti-tumor activity was investigated in B-NDG/hu-PBMC reconstructed mice implanted with NCI-H929. In addition, the potential toxicity profile of LBL-043 was examined in a non-GLP dose range finding (DRF) study in cynomolgus monkeys via IV infusion, once a week for a total of 4 doses. Results: The binding affinity of LBL-043 to LILRB4 and CD3 protein was 0.724 nM and 35.7 nM, respectively. In CD3 reporter gene assays, LBL-043 could activate the NFAT signaling only in the presence of LILRB4+ cells. And LBL-043 was shown a potent TDCC activity in different LILRB4 expression level MM cells. In NCI-H929 CDX mouse model, LBL-043 was shown significant anti-tumor activity. DRF Study in Drug Toxicology Assessments demonstrated that LBL-043 has good safety profile in cynomolgus monkeys. There were no LBL-043 related changes of cardiac parameters, clinical chemistry, hematology, urinalysis, organ weight and gross pathology (except for decreased size of thymus). Conclusion: LBL-043, a novel bispecific antibody targeting CD3 and LILRB4, redirecs T cells to effectively kill MM cells by simultaneously binding MM cells and T cells. It is also shown a great anti-tumor efficacy in NCI-H929 CDX mouse model. DRF of LBL-043 (0.2∼10 mg/kg) in cynomolgus monkeys was well tolerated, and HNSTD was 10 mg/kg. These data for the first time show LBL-043 as a promising therapeutic bispecific antibody for the treatment of MM patients. Citation Format: Yujia Dang, Xiao Huang, Yurong Qin, Xiaoya Liu, Min Chen, Duqing Jiang, Guojin Wu, Mi Ye, Jianming Sun, Hong Ling, Shoupeng Lai, Xiaoqiang Kang, Jordan Zhu. LBL-043, a novel LILRB4xCD3 T cell engager, for the treatment of relapsed/refractory multiple myeloma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 7323.

To develop a rapidly dissolving porous particle formulation of paclitaxel without Cremophor EL that is appropriate for quick intravenous administration. A rapidly dissolving porous particle formulation of paclitaxel (AI-850) was created using spray drying. AI-850 was compared to Taxol following intravenous administration in a rat pharmacokinetic study, a rat tissue distribution study, and a human xenograft mammary tumor (MDA-MB-435) model in nude mice. The volume of distribution and clearance for paclitaxel following intravenous bolus administration of AI-850 were 7-fold and 4-fold greater, respectively, than following intravenous bolus administration of Taxol. There were no significant differences between AI-850 and Taxol in tissue concentrations and tissue area under the curve (AUC) for the tissues examined. Nude mice implanted with mammary tumors showed improved tolerance of AI-850, enabling higher administrable does of paclitaxel, which resulted in improved efficacy as compared to Taxol administered at its maximum tolerated dose (MTD). The pharmacokinetic data indicate that paclitaxel in AI-850 has more rapid partitioning from the bloodstream into the tissue compartments than paclitaxel in Taxol. AI-850, administered as an intravenous injection, has been shown to have improved tolerance in rats and mice and improved efficacy in a tumor model in mice when compared to Taxol.

Pharmacokinetics of 40 kDa Polyethylene glycol (PEG) in mice, rats, cynomolgus monkeys and predicted pharmacokinetics in humans

BOF-4272, (+/-)-8-(3-methoxy-4-phenylsulfinylphenyl) pyrazolo[1,5-a]-1,3,5-triazine-4(1H)-one), is a new drug intended for the treatment of hyperuricemia. This report describes the pharmacokinetics and detailed metabolic pathways of BOF-4272 in the cynomolgus monkey, which were investigated using the metabolites found in plasma, urine, and faeces after intravenous and oral administration. M-4 was the main metabolite in plasma after intravenous administration. M-3 and M-4 were the main metabolites in plasma after oral administration. The Cmax and AUC(0-t) of M-4 were the highest of all the metabolites after intravenous administration. The Cmax and AUC(0-t) of M-3 were the highest of all the metabolites, and those of M-4 were the second highest, after oral administration. M-4 and M-3 were the main metabolites detected in urine and faeces, respectively, after intravenous administration, with M-4 and M-3 at 47.2% in urine and 19.1% in faeces, respectively, within 120 h after administration. M-4 was the only metabolite detected in urine after oral administration, at about 5% within 120 h after administration. M-3 was detected in faeces at 17.0% within 120 h after oral administration. These results suggest that, in the cynomolgus monkey, BOF-4272 is rapidly biotransformed to a main metabolite (M-4, a sulphoxide-containing metabolite of BOF-4272) and that M-4 is mainly excreted in urine and possibly also in bile, with subsequent conversion to M-3 by the intestinal flora. It is expected that the biotransformation of BOF-4272 would be similar in healthy human volunteers.

Nemvaleukin alfa (nemvaleukin, ALKS 4230) is a novel cytokine created by the fusion of circularly permuted interleukin-2 (IL-2) to the IL-2Rα subunit of the IL-2 receptor (IL-2R) complex that confers selectivity for the intermediate-affinity IL-2R expressed on CD8+ T cells and natural killer (NK) cells. The pharmacokinetics and selective pharmacodynamic properties of nemvaleukin have been demonstrated using in vitro and in vivo mouse models. The pharmacokinetic/pharmacodynamic effects of nemvaleukin on immune cell subtypes were evaluated in cynomolgus monkeys after intravenous and subcutaneous administration to inform dose selection and predict pharmacodynamic effects in humans. Male drug-naïve cynomolgus monkeys (N = 15) were administered either single-dose (0.3 mg/kg i.v.; 0.3 mg/kg or 1.0 mg/kg s.c.) or repeated doses (0.1 mg/kg i.v. on days 1-5 or 0.5 mg/kg s.c. on days 1 and 4) of nemvaleukin. Serial blood samples were collected for pharmacokinetic assessment, immunophenotyping by flow cytometry, and profiling of serum cytokines. Repeat-dose subcutaneous administration of nemvaleukin with less frequent dosing resulted in total systemic exposure and trough serum concentrations comparable to those seen with intravenous administration, with lower peak serum concentrations. Transient elevation of interferon-γ and IL-6 peaked at 2 and 8 hours after intravenous and subcutaneous administration, respectively. Selective expansion of immunoprotective central memory, effector memory, terminal effector CD8+ T cells, and CD56+ NK cells, and minimal expansion of immunosuppressive CD4+CD25+FoxP3+ regulatory T cells was observed after both intravenous and subcutaneous administration. These data support the ongoing clinical evaluation of intravenous and subcutaneous nemvaleukin. SIGNIFICANCE STATEMENT: Administration of the novel interleukin-2 receptor agonist nemvaleukin alfa to cynomolgus monkeys resulted in selective expansion of immune effector cells, including CD8+ T and natural killer cells with minimal effects on immunosuppressive CD4+ regulatory T cells, confirming the design of nemvaleukin and highlighting its potential as a cancer immunotherapy. Subcutaneous administration of nemvaleukin achieved systemic exposure and immunostimulatory effects similar to those observed after more frequent intravenous dosing and may represent a practical alternative in a clinical setting.

BackgroundCPGJ701 is a recombinant humanized anti-human epidermal growth factor receptor-2 (HER2) monoclonal antibody-derivative of the cytotoxic agent maytansine (DM1) conjugate for the treatment of HER2-positive metastatic breast cancer. Tissue cross-reactivity (TCR) studies of CPGJ701 in a complete panel of normal human, cynomolgus monkey and Sprague-Dawley were performed to provide evidence for selecting animal species for use in preclinical toxicity studies and predicting primary target organs and clinical adverse drug reactions (ADRs).MethodsTCR studies were carried out to evaluate the distribution of reactivity and the TCR of CPGJ701 in paraffin sections of 32 tissues and/or organs (such as the heart, lung, liver, and kidney) from at least three unrelated normal human, cynomolgus monkey and Sprague-Dawley rat donors. The TCR of CPGJ701was detected by one-step immunohistochemical method using 50 µg/mL biotin-labeled CPGJ701 as the primary antibody. Moreover, a negative biotin-labeled human IgG control group, a blank phosphate-buffered saline (PBS) control group, and a positive human breast cancer tissue control group were also used to exclude false positive and false negative results. The specific positive binding and distribution of reactivity of CPGJ701 were detected in the human breast cancer tissue and 32 tissues from normal humans, cynomolgus monkeys and Sprague-Dawley rats under a microscope.ResultsThe TCR of CPGJ701 in humans and cynomolgus monkeys was highly consistent but showed some differences compared to the TCR of CPGJ701 in Sprague-Dawley rats. The binding of CPGJ701 to target tissues, such as the liver, adrenal gland, thyroid, fallopian tube, spinal cord and skin, was observed in humans and cynomolgus monkeys but not in Sprague-Dawley rats. Specific binding to the placenta was only found in Sprague-Dawley rats. The cell types to which CPGJ701 specifically bound, including epithelial cells, cardiomyocytes and nerve cells, were identical in humans, cynomolgus monkeys and rats.ConclusionsThe TCR of CPGJ701 was in accord with the targeting characteristics of the humanized anti-HER2 monoclonal antibody. The consistency of CPGJ701 binding to human and cynomolgus monkey tissues indicated that the cynomolgus monkey is a relevant animal species for evaluating the preclinical safety of CPGJ701. The targeting (binding site) of CPGJ701 in Sprague-Dawley rats indicated that it is also a useful animal species for evaluating antibody-dependent toxicity and non-antibody-dependent toxicity. In conclusion, these TCR studies of CPGJ701 could provide information for selecting relevant animal species for nonclinical studies and predicting clinical ADRs.

Nonclinical safety, tolerance and pharmacodynamics evaluation for meplazumab treating chloroquine-resistant Plasmodium falciparum

BackgroundMSLN is a membrane protein overexpressed by many cancers with limited expression in normal tissues. One challenge in developing MSLN-targeted therapies has been the presence of a shed MSLN sink...

Evaluation and Modification of a Physiologically Based Model of Lead Kinetics Using Data from a Sequential Isotope Study in Cynomolgus Monkeys

Nonclinical safety evaluation of erenumab, a CGRP receptor inhibitor for the prevention of migraine

Evaluation of the Pharmacokinetics, Immunogenicity, and Safety of XmAb®2513 in the Ongoing Study XmAb2513-01: A Phase 1 Study of Every Other Week XmAb2513 to Evaluate the Safety, Tolerability, and Pharmacokinetics in Patients with Hodgkin Lymphoma or Anaplastic Large Cell Lymphoma

The pharmacokinetics of SCH 27899, a novel oligosaccharide compound of the everninomicin class with excellent activity against gram-positive strains, was studied with mice, rats, rabbits, and cynomolgus monkeys following intravenous administration as SCH 27899-N-methylglucamine-hydroxypropyl beta-cyclodextrin. Concentrations of SCH 27899 in mouse serum, rat plasma, and rabbit serum were determined by a high-pressure liquid chromatography method on a poly(styrene-divinyl benzene) column, and those in monkey plasma were determined by a paired-ion chromatographic method. Plasma and serum concentrations of SCH 27899 exhibited a biexponential decline in all species following intravenous administration. The half-lives at beta phase were 3.0 to 7.9 h in mice, rats, and rabbits and 24 h in cynomolgus monkeys. There was a linear relationship between the area under the curve extrapolated to infinity [AUC(I)] in mice and dose. Rabbits also exhibited dose proportionality in AUC(I). However, in rats, increasing the dose from 3 to 60 mg/kg of body weight resulted in a 49-fold increase in AUC(I). When the species was changed from mouse to rat, rabbit, or cynomolgus monkey, AUC(I) increased, whereas clearance (CL) decreased. It was concluded that the pharmacokinetics of SCH 27899 in animals varied with species; CL was the highest in mice and rats, followed by rabbits and cynomolgus monkeys.