Liver malignant tumors (LMTs) have recently been reported as severe and life-threatening adverse drug events associated with drug-induced liver injury (DILI). DILIs are the most common adverse drug event and can cause the withdrawal of medicinal products or major regulatory action. To reduce the attrition rate and cost of drug discovery, various quantitative structure–toxicity relationship models have been proposed to predict the probability of a DILI based on the chemical structure of a drug. However, there are many unresolved issues regarding the predictors of LMT-inducing drugs, and biologically interpretable prediction models for LMT have not been developed. Here, we constructed prediction models for whether a drug is LMT-inducing based on the activity of molecular initiating events (MIEs), which are biologically interpretable features and are defined as the initial interaction between a molecule and biosystem. We then constructed five machine learning models (i.e., LightGBM, XGBoost, random forest, neural network, and support vector machine) and evaluated their predictive performances. LightGBM achieved the best performance among the tested models. The MIEs making the highest contribution to the model construction for drug-induced LMT were inducement of Enhanced Level of Genome Instability Gene 1 (human ATAD5), nuclear factor-κ B, and activation of thyrotropin-releasing hormone receptor. These results support the previous literature and can be related to the mechanism onset of drug-induced LMT. Our findings may provide useful knowledge for drug development, research, and regulatory decision-making and will contribute to building more accurate and meaningful DILI prediction models by increasing understanding of biological predictors.

In the event of an overdose, the pharmacokinetics of the drug may be altered, resulting in an unexpectedly rapid increase in blood and tissue drug concentrations. Because central nervous system (CNS)-acting drugs are the major cause of hospitalization for overdose, brain concentrations, which are closely related to the development of acute psychotropic symptoms, would be important. However, due to the lack of an appropriate model for overdose, it is difficult to predict the CNS symptoms of patients with acute poisoning. To clarify the toxicokinetics during intoxication with CNS-acting drugs, we investigated the relationship between the dose and concentrations in the blood and brain in mice. Therapeutic or toxic doses of phenobarbital, flunitrazepam, imipramine, and amoxapine were administered intraperitoneally to mice. Serum and extracellular fluid of the brain were collected up to 24 hr after administration and analyzed using LC-MS/MS to determine the pharmacokinetic parameters in the serum and brain. A comparison of the four psychotropic drugs showed that the toxicokinetics of amoxapine in the blood and brain are clearly different from others, with the brain concentrations being specifically highly susceptible to increase during dose escalation. These results are consistent with the CNS-related symptoms observed in amoxapine overdose. Therefore, the methodology of the current study could be useful for predicting CNS toxicity during psychotropic drug poisoning.

A reactive oxygen species (ROS) assay is an in chemico photoreactivity test listed in ICH S10 guideline and OECD Test Guideline No. 495. We currently utilize the ROS assay to assess the photosafety of cosmetic ingredients. We have recently confronted a problem that there was an absorbance increase of blank assessing superoxide anion generation after irradiation, whereas this did not occur in the negative control (sulisobenzone), leading to a dissatisfaction of the acceptance criteria. Therefore, we aimed to investigate the causes and find countermeasures. No significant effects of impurities and manufacturer differences of sodium phosphate and DMSO on blank absorbance increases were observed. In contrast, when Cu²⁺ was added to the buffer, the increase of blank absorbance after irradiation did not occur. We then confirmed the dose-response relationship and found that adding 0.1 μM of Cu²⁺ (corresponding to 6 ppb of Cu²⁺) was sufficient in suppressing the blank absorbance increase, suggesting the need of Cu²⁺ supplementation to the buffer. Finally, we confirmed that the ROS assay using the buffer supplemented with 0.1 μM of Cu²⁺ obtained stable test results by using 17 proficiency chemicals listed in TG 495. Our results suggest that the modified ROS assay protocol would be useful for obtaining stable test results.

Transient Receptor Potential Melastatin 8 (TRPM8) is a calcium-permeable, non-selective cation channel of the transient receptor potential superfamily, required for the transduction of moderate cold temperatures. TRPM8 is also known to regulate proliferation of prostate, pancreatic, breast, and melanoma carcinoma cells. Here, we examined a key factor in the regulation of TRPM8-mediated proliferation of epidermal cells, which are directly affected by cold temperatures. Experiments involving knockdown and ectopic expression of TRPM8 in normal keratinocyte HaCaT and squamous carcinoma SAS cells suggest that TRPM8 inhibits cell proliferation by upregulating the expression of cyclin-dependent inhibitor p21/Cip1. Whereas these findings were observed in the absence of an endogenous agonists, additions of the synthetic TRPM8 agonist icilin reduced DNA synthesis in HaCaT cells but stimulated that in SAS cells by altering p21/Cip1 levels in a TRPM8-independent manner, indicating that icilin poses a risk of stimulating carcinoma cell proliferation. Unexpectedly, the TRPM8 blocker, used for the treatment of overactive bladder and bladder pain, N-(3-aminopropyl)-2-{[(3-methylphenyl) methyl] oxy}-N-(2-thienylmethyl) benzamide hydrochloride salt (AMTB) reduced DNA synthesis by upregulating p21/Cip1 expression. However, another TRPM8 blocker, N-(4-Tertiarybutylphenyl)-4-(3-chloropyridin- 2-yl) tetrahydropyrazine-1 (2H)-carbox-amide (BCTC), stimulated DNA synthesis by downregulating p21/Cip1 expression, indicating that it may pose a risk of carcinogenesis associated with dysregulated cell cycles when used to treat overactive bladder and bladder pain.