In vitro safety profiling for Medchem Support

Undesirable off-target activity profiles could hinder or halt the development of candidate drugs or even lead to market withdrawal if discovered after a drug is approved. To tackle this issue early in drug development, in vitro pharmacological profiling has been widely used by pharmaceutical companies to identify off-target effects. This low cost “early” strategy has proved to help identify potential ADRs (adverse drug reactions) and increase chances of success for drug approval and safety.

WuXi AppTec offers a in vitro mini-Safety Panel Service to be used for early pharmacological profiling of off-target and adverse effects of lead compounds or PCCs in the drug development stage. Data is also useful for drug candidate SAR design. Assay targets in the panel are selected to form a minimal panel, recommended by scientists from four major pharmaceutical companies (AZ, GSK, Novartis, Pfizer)1, to provide broad and diverse early in vitro safety assessments of testing compounds. Compounds can be screened by radioligand binding, biochemical and/or functional assays.

GPCRs

Receptor (Human)

Main organ class or system

Radioligand Binding

Finction Assay (Ca²⁺ Assay)

Adenosine receptor

Ad2A

CVS, CNS

Adrenergic receptors

Alpha1A

CVS, GI, CNS

alpha2A

CVS, CNS

Beta1

CVS, GI

Beta2

Pulmonary, CVS

Cannabinoid receptor

CB1

CNS

CB2

Immubne

Cholecytokinin receptor

CCKa

GI

Dopamine receptors

D1

CVS, CNS

D2

CVS, CNS, endocrine

Endothelin receptors

ETa

CVS, development

Histamine receptor

H1

CVS, immune

H2

GI, CVS

5-Hydroxytryptamine receptor

5HT1A

CNS, endocrine

5HT1B

CVS, CNS

5HT2A

CVS, CNS

5HT2B

CVS, pulmonary, development

Muscarinic receptor

M1

CNS, GI, CVS

M2

CVS

M3

GI, pulmonary

Opioid receptor

Op-delta

CNS, CVS

Op-kappa

GI, CNS, CVS

Comprehensive introduction brochure: Click for WuXi Early in Vitro Mini Safety Panel 

Assays and representative data summary: Click for WuXi Early in Vitro Mini Safety Panel Assay and Data Summary 


In Vitro Tox Assays for Medchem Support

Toxicity is the major cause of drug candidate failure in the preclinical and clinical development stages as well as the major reason for the withdrawal of approved drugs from the market. While a toxicity test has been traditionally completed in the preclinical phase, in vitro toxicity studies in the early drug discovery stages could significantly reduce failures at a later stage and prevent economic loss.

WuXi AppTec offers a panel of in vitro toxicity assays designed to identify compound toxicity in the early drug discovery stage. By utilizing cutting-edge technologies including conventional and automated patch clamp and high content screening (HCS), we provide the high-quality data quickly and cost-effectively. Applying these assays to your lead ID and optimization strategy helps to make a more thorough analysis of the severity and specificity of toxicity. Then BTO guide candidate compounds through the planning and execution of downstream in vivo toxicity and efficacy tests.

Cardiotoxicity

  • Cardiac ion channel safety panel (CIPA compliant) with manual and automated patch-clamp: including hERG, hCav1.2, hNav1.5 (early and late), hKCNQ1/mink, hKv4.3/KChIP2.2, hKir2.1 channels
  • Human stem cell derived cardiomyocytes cardiotoxicity testing with MEA

General Cytotoxicity

  • Cell viability Assay: CellTiter-Glo® (Promega) or HCS assay
  • Apoptosis assay
  • CellTox™ Green cytotoxicity assay (Promega)
  • ApoTox-Glo™ Triplex Assay (Promega)

Mitochondrial Toxicity

  • Mitochondrial Membrane Potential Assay
  • Mitochondrial Reactive Oxygen Species (ROS) Assay
  • MitoXpress® Xtra OCR Assay (HS method) (Luxcel)
  • MitoXpress® Cellular Energy Flux OCR/ECAR Assay from (Luxcel)
  • Glucose/Galactose Assay
  • MitoBiogenesis Assay

Lipotoxicity

  • Phospholipidosis and steatosis assays

Hepatotoxicity with HepG2 or Primary Human Liver Cell Culture

  • Selected general cytotoxicity and mitochondrial toxicity assays listed above

Nephrotoxicity with HK-2 Cells

  • Selected general cytotoxicity and mitochondrial toxicity assays listed above

Genotoxicity

  • Ames assay
  • In vitro micronucleus assay

Photo Toxicity

  • In vitro 3T3 NRU Phototoxicity test
Fig 1. A hERG current recorded on QPatch, B hiPSC-vCMs recording on MEA system.
Fig 2, Mitochondrial membrane potential assay using HepG2 cells.
Fig 3. Nephrotoxicity assay with HK-2 cells. 
Fig 4. In vitro 3T3 NRU phototoxicity test

Early Stage Safety Assessment

Our safety pharmacology group profiles compounds for potential adverse drug reactions at various R&D stages thereby helping to deliver high-quality drug candidates faster and at lower cost.

Integrated Cardiovascular Safety Assessment

  • Cardiac ion channel safety panel with manual and automated patch-clamp: all CIPA recommended channels including hERG, hCav1.2, hNav1.5 (early and late), hKCNQ1/mink, hKv4.3/KChIP2.2, hKir2.1 channels
  • Human stem cell derived cardiomyocytes cardiotoxicity testing with MEA
  • Action potential recordings in heart muscle and/or Purkinje fibers
  • Telemetry ECG, blood pressure and core body temperature recordings in freely moving animals

In Vivo CNS Safety Assessments

  • Irwin or FOB test
  • Rotarod test
  • Locomotors activity
  • Grip strength

Off-Target Panel Screening

  • >90 Kinase Assay 
  • 100 GPCRs (For GPCR list, please click here)

In Vitro Safety and Selectivity Profiling 

Identify off-target based adverse drug reactions (ADR) at the early stage of drug discovery. Such information may help guide compounds of interest through downstream in vivo tests for potential liabilities and reduce attrition of drug candidates and even the withdrawal of approved drugs.

Led by a group of experienced scientists in drug discovery, our safety panel team offers in vitro mini-Safety Panel Service for early pharmacological profiling of off-target and adverse effects of lead compounds or PCCs in the drug development stage. The data is also useful for drug candidate SAR design. Assay targets in the panel are selected to form a minimal panel, recommended by scientists from four major pharmaceutical companies, to provide broad and diverse early in vitro safety assessments of testing compounds. Compounds can be screened by radioligand binding, biochemical and/or functional assays.

GPCR (Human)

Ion Channel (human or rat*)

Transporter NHR (human)

Enzyme (human)

Ad2A

L-Ca²⁺

DAT

LCK

Alpha1A, 2A; Beta 1, 2

hERG

5HTT

PDE3A

CB1, 2

Kv* or hKCBQ1 + MinK

NET

PDE4D

CCKa

hNav1.5

AR

COX1

D1, 2

5-HT3a

GR

COX2

ETa

nAChR, ⍺7

ACHE

H1, 2

GABAₐ, ⍺1

MAO-A

5HT1A, 2A, 1B, 2B

NMDAR*

M1, 2, 3

Op-delta, Kappa, mu

V1a

Table. Comprehensive targets (44) included in WuXi in vitro mini-safety panel