Soft Spot Identification Service

Soft Spot Identification

The successful development and marketing of a new drug requires a complex process, including the selection of drug targets, the screening of lead compounds, the determination of clinical candidate compounds, the confirmation of clinical efficacy, and the safety evaluation of drugs. When the target screening obtains a molecular structure with in vitro pharmacodynamic activity, it is hoped that it can have the appropriate exposure amount and duration of action in vivo, and the rapid metabolic clearance of the drug tends to reduce the exposure of the drug. To improve the properties of rapid metabolism, it is important to be able to identify the metabolic sites of compound molecules and protect them. The chemical groups or individual atoms of compounds that are prone to metabolic reactions in vivo/in vitro are called "soft spots of metabolism".

High-throughput Metabolic Soft Spot Identification Platform

Our Service

The traditional metabolite identification process not only requires long-term LC-MS analysis to ensure chromatographic separation between metabolites and biological matrix components, but also requires a lot of manual data interpretation. Creative Biolabs provides a rapid, high-throughput method for identifying metabolic soft spots to meet throughput requirements in the early stages of drug discovery.

Fig.1 Our soft spot identification service process.

Analytical Techniques:

LC-MS methods can shorten the time to test samples.
UPLC ensures the retention and separation of compounds and their metabolites in a short period.
High-resolution mass spectrometry (HRMS) for sample detection ensures more accurate mass-to-charge ratio information.
Multiple data acquisition modes (Full MS/dd-MS2/MSE) and rich ion filtration are used to find metabolites and provide accurate fragmentation information for attribution of metabolic soft spots.

Published Data

Objectives

In this study, a simple, effective, and high-throughput metabolic soft-spot identification assay was developed and validated. Determine the metabolic soft spots of test compounds in liver microsomes by using univariate incubation time and LC/UV/MS.

Method

Fig.2 Experimental flow. (Zhu et al., 2022) Fig.2 Experimental flow of this study.1

Results

Metabolic Soft-Spot Determination Using LC/UV/Q-TOF
Fig.3 Metabolic soft-spot identification of four metabolites using LC/UV/Q-TOF.¹	Quantitative and qualitative analysis of four model compound samples in HLM incubation was performed using LC/UV/Q-TOF to determine their metabolic soft spots. As shown in Fig.3 A, C, and E, they all present a single dominant component.
Fig.4 MS/MS spectra of the major metabolites of the two compounds acquired by LC/Q-TOF.¹	Fig.4 shows the MS/MS spectra of the major metabolites of the two compounds obtained by LC/Q-TOF. Based on the results of full-scan MS and MS/MS spectroscopy data, the metabolic soft spots of four model compounds in the HLM incubation were identified.

Metabolic Soft Spot Determination by LC/UV/Qtrap
Fig.5 Metabolic soft spot identification of a compound in HLM by LC/UV/Qtrap.¹	In this study, an LC/UV/Qtrap method was developed for the identification of metabolic soft spots for these four compounds. The results in Fig.5 show the LC/UV curve for one of the compounds.

In the face of the rapid screening needs in the early stage of drug development, our metabolic soft spot identification platform is based on ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-UV-HRMS) technology, professional data processing software and rich research experience, and continuously improves speed and throughput on the premise of ensuring quality.

If you have any soft spot identification needs, please contact us!

Reference

Zhu, Yanlin et al. "High-throughput metabolic soft-spot identification in liver microsomes by LC/UV/MS: application of a single variable incubation time approach." Molecules. (2022) 27,22 8058.

For Research Use Only | Not For Clinical Use