When does MIST study need to be carried out？

When a metabolite (especially Phase I metabolite) in human steady state plasma accounting for more than 10% AUC, it is necessary to compare the exposure of the major metabolite in steady state plasma in human and in toxicological species to evaluate whether the safety of the major metabolite has been adequately assessed or not.

When is the suitable time for MIST?

MIST studies are conducted during Phase I clinical study MAD stage.

What samples are used for MIST?

Plasma samples at steady state under clinical therapeutic dose are selected to conduct MIST study. Steady state plasma samples in toxicological species under NOEAL are selected to compare with that in humans.

Metabolites in Safety Testing (MIST)

Compare the exposure of the major metabolites in steady state plasma in human and in toxicological species when a metabolite (especially Phase I metabolite) in human plasma accounting for more than 10% AUC to evaluate whether the safety of the major metabolites has been adequately assessed or not.

REQUEST A QUOTE

Overview
Assays
Study Strategies
Case Study
Regulatory Guidance
Experience
Instruments and Software
FAQs
Related Resources
Related Services

MetID

Connect with us

Overview

WuXi AppTec DMPK can provide MIST research services and guide analytical decisions in a progressive manner based on experimental results. Our team has over 15 years of industry experience and includes dozens of experienced metabolic researchers. We have established a state-of-the-art platform equipped with various models of high-resolution mass spectrometry, including the Orbitrap Eclipse™ Tribrid™, Orbitrap Exploris™ 480, Q Exactive™ (QE), Q Exactive™ PLUS, Q Exactive™ HF, and VION™ IMS Qtof and other models of high-resolution mass spectrometry. Our research services cover the drug discovery stage from lead compound optimization selection to drug clinical research stage, as well as metabolic product analysis and identification using radioactive isotope tracing. We have supported the completion of over a thousand global application projects, with nearly a hundred clinical metabolite identification projects. We can complete thousands of metabolite identification studies annually.

Learn More

Assays

MetID in toxicology animals
- Steady-state phase in toxicology multiple ascending dosed animals
- AUC pooled plasma
- Analysis by LC-UV-HRMS
- Metabolite profiling and identification
- Determination of the relative abundance of parent and metabolites in plasma
MetID in human
- Clinical MAD (multiple ascending doses) in steady state
- AUC pooled plasma
- Analysis by LC-UV-HRMS
- Metabolite profiling and identification
- Determination of the relative abundance of parent and metabolites in plasma
Compare the exposure of metabolites
- Major metabolites with relative abundance ≥10% from human
- Compare the peak area of the major metabolites between human and animal
- Calculate the exposure ratio
- Assess whether the exposure of the metabolites in animal can cover human

Start your study

Study Strategies

After the drug enters the human body, the produced metabolites are all exogenous substances for the human body. When the exposure to a metabolite at a steady state exceeds 10% of the total exposures of all drug-related components in human, the safety of this metabolite should be evaluated in toxicological studies. Drug metabolites in plasma can be searched and identified using high-resolution mass spectrometry combined with relevant data processing software, as well as the relative abundances of the detected metabolites. For major metabolites, further comparison of its exposure to human and toxicological species should be determined. Results from the study can help to evaluate if the selected toxicological species are appropriate.

Case Study

A pooled plasma sample from human after repeated administration of a drug candidate is prepared by an AUC method, and then analyzed by LC-UV/HRMS. As shown in Figure 1 and Table 1, five metabolites are detected, identified and semi-quantitatively determined. Both M4 and M5 account for more than 10% of the exposure of the total drug-related components in human at steady state. Due to no metabolite standards, the peak areas of the human metabolites in the AUC-pooled human and animal plasma samples are compared (Figure 2). Results from the analysis can confirm whether the exposure of high proportion metabolites in toxicological animal species is higher than in human. To avoid matrix interference, blank human and animal plasma were added to real animal and human plasma (Figure 2). The formula in Figure 2 was used to evaluate the exposure of metabolites in human and animal plasma.

Learn More
- Metabolite profiles of parent drug and its metabolites in human plasma at steady state
  
  Figure 1
- Relative abundance of parent drug and its metabolites in human plasma at steady state
  
  Table 1
- A) Mixed method to overcome matrix effect
  
  Figure 2
- B) Formula for calculation of exposure multiple
  
  Figure 2

Regulatory Guidance

Safety Testing of Drug Metabolites Guidance for Industry

Experience

10+

Years of experience
80%

Successfully supported NDA submissions for FDA and NMPA
10+

MIST studies

Instruments and Software

Instruments
Software

- Thermo Orbitrap Eclipse™ Tribrid™
- Thermo Orbitrap Exploris™ 480
- Thermo Q-Exactive™ HF
- Waters VION™ IMS QTof
- Thermo Q-Exactive™
- Thermo Q-Exactive™ Plus
- Therno LTQ Orbitrap XL
- Waters Xevo®G2 QTof
- Solid scintillation counter: off-line detection of radioactivity
- Liquid scintillation counter: total radioactivity detection
- On-line detection of radioactivity
- Thermo Scientific™ Compound Discoverer™
- Thermo Scientific™ Mass Frontier™
- Thermo Scientific™ Metworks™
- Waters MetaboLynx^TM
- Mass Analytical Mass-MetaSite
- Waters UNIFI^®
- Thermo Scientific™ Biopharma Finder™

FAQs

When does MIST study need to be carried out？

When a metabolite (especially Phase I metabolite) in human steady state plasma accounting for more than 10% AUC, it is necessary to compare the exposure of the major metabolite in steady state plasma in human and in toxicological species to evaluate whether the safety of the major metabolite has been adequately assessed or not.
When is the suitable time for MIST?

MIST studies are conducted during Phase I clinical study MAD stage.
What samples are used for MIST?

Plasma samples at steady state under clinical therapeutic dose are selected to conduct MIST study. Steady state plasma samples in toxicological species under NOEAL are selected to compare with that in humans.

Related Resources

Browse All Metabolites in Safety Testing (MIST) Resources

Related Services

Stay Connected

Keep up with the latest news and insights.

Email address*

* Please check the filled content
First name*

* Please check the filled content
Last name*

* Please check the filled content
Company*

* Please check the filled content

By clicking submit, you consent to WuXi AppTec DMPK collecting and processing the information you provide for our internal purposes, in accordance with our privacy policy.

* Please agree to the Privacy Policy

Submit

Metabolites in Safety Testing (MIST)

Overview

Assays

MetID in toxicology animals

MetID in human

Compare the exposure of metabolites

Study Strategies

Case Study

Regulatory Guidance

Experience

Instruments and Software

FAQs

Related Resources

Optimisation of an In Vitro Plated Monkey Hepatocyte Model and Comparative Metabolite Profiling of a GalNAc-Conjugated siRNA, siRNA01, in Monkey Liver Homogenate and Hepatocytes

Probing Drug and Metabolite-Covalent Binding with a Non-Radiolabeled Workflow: Osimertinib as an Example

Precision Metabolite Identification & Biotransformation for Drug R&D-WuXi AppTec DMPK

New Book Release: Drug Metabolism and Pharmacokinetics: Frontiers, Strategies, and Applications | WuXi AppTec DMPK

Rapid Screening and Characterization of Payload-Related Catabolites Derived from ADCs in In Vitro Models by UPLC-UV-HRMS

Decoding DMPK Frontiers for Novel Therapeutics: WuXi AppTec DMPK’s New Book Release

Drug Metabolism and Pharmacokinetics: Frontiers, Strategies, and Applications

Unlocking Reliable In Vitro Metabolic Models for Oligonucleotide Therapeutics with the Latest Publication on DMD

High-Performance, Professional, Innovative: WuXi AppTec DMPK Accelerates Breakthroughs

Comparative Metabolism of a GalNAc-Conjugated siRNA, Inclisiran, Among Various In Vitro Systems and Correlations with In Vivo Metabolism in Rats

Detection and Characterization of In Vitro Payload-Containing Catabolites of Noncleavable Antibody-Drug Conjugates by High-Resolution Mass Spectrometry and Multiple Data Mining Tools

Covalent Drugs DMPK Services

Unlocking Precision in Peptide Metabolite Identification for Next-Gen Drug Development

Insights into Chiral Drug Metabolism and Inversion

Methodologies and Strategies for ADC Biotransformation Studies

WuXi AppTec DMPK: Your Trustworthy Partner

Metabolite Identification in Peptide Drugs and Its Challenges

Comparative Metabolite Profiling and Identification of a GalNAc-Conjugated siRNA, siRNA01, in Plasma Prepared with Various Anticoagulants, Serum, and In Vivo Plasma Using LC-UV-HRMS

Overcoming Challenges in Precise Structural Identification of Target Metabolites

Involvement of Aldehyde Oxidase (AOXs) in Drug Metabolism: Early Prediction and Coping Strategies

Prodrug Approaches for Improved DMPK Characteristics and Their Preclinical Study Considerations

Navigating the Path from IND to NDA

An Analytical Workflow for Polymer Metabolism: PEGylated Lipids Biotransformation with LC-HRMS

Development of a Convenient In Vitro Method for Predicting Metabolism and Disposition of Acrylamide Covalent Drugs in Humans

Effect of pH on Metabolite Profiling and Identification of GalNAc Conjugated siRNA in In Vitro Metabolic System

Metabolite Profiling and Identification of Oligonucleotide in In Vitro Metabolic System

DAR Distribution Determination for Antibody-drug-conjugates (ADCs) by LC-HRMS in In Vitro and In Vivo Studies

Drug Metabolism and Pharmacokinetics (DMPK) Service

One-stop Metabolite Biosynthesis and Structural Characterization

Overcoming the Challenges of Metabolite Identification and Profiling for Developing Oligonucleotides

Metabolite Profiling & Identification (MetID) Services

Oligonucleotide Drugs: Strategies for Metabolism and Metabolite Profiling and Identification

In Vitro-In Vivo Metabolite Profiling and Identification of Oligonucleotide

A New Method to Improve Identification of the Payload-Containing Catabolites of ADCs

How to Address the Challenges of PROTAC Metabolism

How MetID Studies can Improve Safety and Efficacy in PROTAC Drugs

Related Services

Stay Connected