Erse agonist; liquid chromatography andem mass spectrometry; metabolite identification; metabolomicsPharmaceutics 2021, 13, 776. https://doi.org/10.3390/pharmaceuticshttps://www.mdpi.com/journal/pharmaceuticsPharmaceutics 2021, 13,2 of1. Introduction Estrogen-related receptors (ERRs) are orphan nuclear receptors consisting of subunits ERR, ERR, and ERR. ERRs are primarily expressed inside the brain, heart, kidney, and liver, and they play a essential function in regulating cellular metabolism and power homeostasis . ERR is especially involved in metabolic illnesses for example type 2 diabetes mellitus and alcohol-induced oxidative liver injury  triggered by impaired hepatic gluconeogenesis  and insulin signaling . ERR has also been reported to mediate the transcriptional response in cancer . Thus, it is at present thought of a possible therapeutic target for treating metabolic diseases including form 2 diabetes mellitus and cancer [3,6]. ERR features a generic function in regulating power metabolism like other subtypes; having said that, it plays a certain part in embryonic improvement, cell replication and differentiation [7,8]. In unique, ERR expression inside the breast cancer patient tissues was down-regulated in comparison with standard breast tissues . It suggests that ERR may also be a possible therapeutic target. Various ERR-selective inverse agonists have already been created, including GSK5182 , DN200434 , and DY40 . GSK5182 and DN200434 improve sodium iodide symporter function and radioiodine activity in anaplastic thyroid cancer cells in vitro, but only DN200434 (which has far better in vivo pharmacokinetic profiles and biocompatibility than GSK5182) was effective in an in vivo anaplastic thyroid cancer model. DN203368 ((E)-3-[1(4-[4-isopropylpiperazine-1-yl]phenyl)3-methyl-2-phenylbut-1-en-1-yl]phenol, Figure 1) is a 4-hydroxy tamoxifen analog that is definitely an inverse agonist of estrogen-related receptor / (ERR/). Improvement of compound DY40 as an inverse agonist of ERR/ (IC50 , 0.01 ) and DY181 as an inverse agonist of ERR (IC50 , 0.05 ) was reported with only preliminary in vitro data . Using the aim of discovering a novel dual ERR/ inverse agonist, a lead compound, DN203368, was developed. Additional research on examining anticancer effects on many in vitro and in vivo cancer models employing DN203368 as ERR/ ligand are ongoing. Drug metabolism could be the biotransformation method that tends to make xenobiotics additional polar to facilitate their elimination from the physique. The items of metabolism are commonly inactive, but active metabolites may also be generated. In some circumstances, metabolism results in the formation of reactive or toxic metabolites that can have an effect on drug safety . Drugs such as troglitazone, trovafloxacin, bromfenac, and lumiracoxib have already been withdrawn in the market place CCR3 list resulting from adverse effects brought on by toxic metabolites . It is actually thus incredibly significant to understand the metabolism of new drug candidates within the early stages of drug discovery and improvement, and it is also crucial for regulatory agencies to confirm the safety of your drug.Figure 1. Chemical structure of DN203368.Generally, metabolite identification is carried out by comparing the parent drug’s mass fragmentation pattern with 4-1BB web prospective metabolites immediately after liquid chromatography andem mass spectrometry (LC-MS/MS) . This traditional method mostly relies on person understanding and practical experience of drug metabolism and interpretation of massPharmaceutics 2021, 13,3 offragment.