Liquid chromatography-mass spectrometry (LC-MS) bioanalysis technology plays a vital role in early drug development and can promote the process of new drug development. Before the introduction of combinatorial chemistry, many drug candidates from natural products and active compounds were extracted and separated, and their chemical structures needed to be analyzed by chemical process test platforms such as nuclear magnetic resonance, mass spectrometry, infrared, and derivatization or selective chemical degradation. After the introduction of combinatorial chemistry, early drug discovery requires the development of efficient LC-MS bioanalytical technology to support quantitative analysis of bioanalysis.
The drug development process has undergone rapid changes with the emergence of combinatorial chemistry, genomics, proteomics, and bioinformatics, and new drug therapeutic targets have also increased; at the same time, the application of high-throughput screening and ultra-high-throughput screening technologies Greatly increase the number of new chemical entities (NCEs). However, few compounds have finally successfully become drugs through preclinical and clinical trials. Many pharmaceutical developers try to use the results of in vitro experiments to predict the behavior of drugs in the human body by conducting in vitro experiments in the early stages of new drug development to determine whether there is value for further research.
The role of LC-MS bioanalysis technology in the screening of hit compounds
Mass spectrometry is one of the inherent characteristics of compounds. Different compounds have different mass spectra. Except for some isomers, it can provide more information for structural qualitativeness. It is an ideal chromatographic detector and a powerful structural analysis tool. The multi-channel monitoring function of mass spectrometry technology and the excellent separation ability of liquid chromatography technology make LC-MS technology significantly reduce the concentration and purity requirements of detection samples, which is very suitable for the research of drugs with complex active ingredients and low content. Medicilon Bioanalysis Service Department has a professional scientific research team, the analysis laboratory is equipped with advanced equipment, implements comprehensive information management, experimental research meets the requirements of FDA/CFDA GLP standards, and the service content involves pharmacokinetics and pharmacodynamics , Immunogenicity and bioequivalence, etc., to provide customers with the screening and development of small molecule drugs, biological agents, vaccines and biomarkers, as well as preclinical and clinical research.
The use of high-throughput LC-MS bioanalysis technology can test the solubility, membrane permeability or transport, protein binding rate, chemical and metabolic stability of those compounds that have been identified as seedlings. These in vitro experiments can not only test the reliability of virtual screening before synthesis, but also become the basis for selecting compounds worthy of further development. Studies have used LC-MS technology to screen Cardiogenol C-producing strains from 51 Bacillus strains with high throughput .
Cardiogenol C is a diaminopyrimidine compound that induces the differentiation of MHC- (myosin heavy chain)-positive cardiomyocytes derived from embryonic stem cells with an EC 50 value of 0.1 μM. Cardiogenol C hydrochloride is a cell-permeable pyrimidine compound that effectively induces embryonic stem cells to differentiate into cardiac muscle. Researchers use MassHunter software to extract molecular features, and quickly compare them in the Metlin metabolic database based on retention time and precise molecular weight information, and identify metabolites with high throughput. A strain of Bacillus amyloliquefaciens FJAT-17934 Bacillus amyloliquefaciens produced Cardiogenol C was screened and identified. The relative content was 1.02%, the matching degree reached 92.08%, and the retention time was 3.651 7min. This lays a theoretical foundation for the development of Cardiogenol C from microbial sources.
The role of LC-MS in drug efficacy evaluation and candidate drug selection
The compounds selected by high-throughput screening are then evaluated by pharmacological models. If target biochemistry is applied to LC-MS analysis, any targeted pathway or metabolomics method can be used in pharmacological research. High-throughput screening of potential biomarkers. If successful, the selected biomarkers will play a great role in preclinical and clinical research.
LC-MS bioanalysis technology plays a fundamental role in many successful cases of drug discovery. If properly designed, early in vitro studies can determine the compound’s inherent clearance rate in multiple species, and in vitro evaluation improves the use of inherent clearance to predict system clearance efficiency. Through in vitro data, the in vivo metabolic clearance rate of the drug can be estimated, and this pharmacokinetic parameter can be used to further calculate the bioavailability and half-life, and the above parameters are very useful in determining its in vivo pharmacology, toxicological effects, and patient compliance. It is important and has a very important reference value for determining the first dose and frequency of administration in the clinic.
The use of combinatorial chemistry such as cassette administration, that is, the simultaneous administration of multiple compounds, is one of the means to quickly assess the penetration of the drug to the target location. Usually about 20 compounds can be administered simultaneously, but some researchers have tried to administer as many as 100 compounds simultaneously. The specificity of mass spectrometry detection allows simultaneous measurement of many compounds in body fluids and tissues, and rapid screening of drug candidates that can penetrate the site of action. Moreover, the sample pretreatment of LC-MS is simple and generally does not require derivatization or hydrolysis, making the LC-MS combined technology has broad application prospects in drug development.
 High-throughput screening of Cardiogenol C-producing Bacillus based on LC-MS technology [J].