ADME & Bioanalysis
Red Glead Discovery’s ADME & Bioanalysis function supports drug discovery projects by providing lab services designed to analyze compound characteristics. We provide analytical expertise in order to help our clients determine their drug candidates’ suitability for further development.
Why choose us
We offer a variety of in-house bioanalytical and physicochemical services, with both standardized assays and custom analytical services to suit the client’s needs. With years of experience and state-of-the-art chromatography and mass spectrometry capabilities, we are equipped to assist with a range of services. Our competent team is experienced in both quantitative and qualitative analysis for a wide range of small molecules and peptides. We offer multiple techniques for chromatographic separation and detection, with HPLC-MS/MS as standard technique for identification and quantification.
We value close communication with the client and agility in problem-solving as challenges arise along the way. Being involved from the initial screening of physical properties such as solubility and lipophilicity to in vitro metabolism and study design of pharmacokinetic experiments, we offer knowledge encompassing many facets of the drug discovery process.
ADME & Bioanalysis and Integrated drug discovery
We support drug discovery projects from hit-to-lead with background knowledge from both medicinal chemistry and analytical chemistry fields. We work closely with the peptide and medicinal chemistry teams to assist with assessing the physicochemical properties of a candidate drug as well as offering bioanalytical services such as plasma stability, microsome in vitro metabolism and plasma protein binding.
Case 1
An internal integrated drug discovery project presented very low solubility in their initial lead compound series, with a typical buffer solubility of <0.5 µM. Weekly solubility screens throughout the lead optimization phase gave input into the design of new, more soluble molecules. These test/design cycles resulted in improvements of 10-100 fold for the solubility of the final candidates.
The results from the solubility screen assays were followed up monthly by solid solubility assessments of compounds with known solid state (crystalline compounds). Crystallinity assessments were performed with XRPD by a collaborative partner also located at Medicon Village, Lund, Sweden.
Solubility was confirmed to be up to 200 µM in biorelevant fluid, FaSSIF, without losing potency against the target. The low solubility is not likely to limit absorption in humans (MAD calculations).
Case 2
Lead candidate design process based on observed pharmacokinetic properties. Red Glead Discovery’s ADME & Bioanalysis team assisted a client in performing quantitative bioanalysis for a range of peptides in several in vivo studies. The studies were performed continuously with the aim of finding an optimal half-life for a peptide candidate drug. Based on pharmacokinetic parameters calculated from previously tested candidates, the peptides were modified with the aim of reaching improved half-lives. After a few iterative design cycles, candidates were found in the desired half-life window for weekly subcutaneous administration.
Our Approach
The process begins with close communication between our team and the client in order to assess how we can address the projects needs in the most effective way. We spend a considerable amount of the optimizing analytical parameters for a given compound in order to produce a robust and sensitive method of evaluation. The ADME & Bioanalysis team offers a range of services, both standardized assays and custom study set-ups:
Solubility screen
Solubility assessment up to 100 µM in phosphate buffer pH 7.4 (with 1% DMSO)
Lipophilicity (logD pH 7.4)
Distribution coefficient between octanol and phosphate buffer pH 7.4 is determined in a classical shake-flask experiment.
Solid solubility
Solubility assessment from solid material, preferable crystallin material. Used as follow-up method for screen solubility for selected compounds.
FaSSIF solubility
Solubility assessment in Fasted state simulated intestinal fluid at pH 6.5, +37°C.
Customized solubility
Solubility assessment from solid material. Generally used during development of formulations for early in vivo experiments. Highly adaptable assay in terms of incubation conditions such as media and pH selection, maximum concentration etc.
Chemical stability
Stability assessment for solid material or solutions. Highly adaptable assay in terms of incubation conditions such as media and pH selection, concentration, timepoints, stress factors (temperature, light exposure, humidity) etc.
PAMPA
(Parallel Artificial Membrane Permeability Assay)
Assessment of passive permeability with a filter plate pre-coated with structured layers of phospholipids.
General: Quantitative bioanalysis by LC-MS/MS
Determination of analyte concentrations in biological samples (plasma, whole blood, tissue, urine, faeces). Analysis of one or multiple analytes in the same sample.
Metabolic in vitro stability in microsomes
A standardized assay, run in 96-well format. The metabolic decline is followed at 6 time points, 45 minutes. The relative peak areas are detected by LC-MS/MS (mouse, rat, dog, pig, human).
Metabolic in vitro stability in hepatocytes
A standardized assay, run in 96-well format. The metabolic decline is followed at 6 time points, 60 minutes. The relative peak areas are detected by LC-MS/MS (rat, human).
Plasma Protein Binding
Assessment of Plasma Protein Binding through rapid equilibrium dialysis (mouse, rat, pig, dog, human)
Plasma stability, in vitro
Highly adaptable assay in terms of incubation conditions such as plasma species, compound concentration, timepoints, stress factors (temperature) etc.
Enzyme stability
Custom stability assay set-up of any commercially available enzyme.
We’re here to support during the entire cycle of your pharmacokinetic (PK) study. At Red Glead Discovery, we do not provide in vivo testing ourselves but have close collaborations with several CROs located at Medicon Village in Lund conducting these studies. We can help you with study planning and logistics to avoid unwanted disruptions or delays in your study.
We offer:
- Pre-clinical formulation design: stability and solubility testing in known well-tolerated drug vehicles prior to in vivo study
- Preparation of formulation and delivery to the study subcontractor in close connection to project start
- Reception of plasma or tissue samples from the study subcontractor
- Bioanalysis of the samples
- Pharmacokinetic calculations and interpretation of results
Adsorption investigation
Assessment of potential sources of analyte adsorption
Equipment
Sciex QTRAP 4500 LC-MS/MS system with a Shimadzu UPLC-UV
Sciex TQ 6500+ LC-MS/MS system with Exion UPLC
Sciex QTOF X500-B with Exion UPLC
Agilent 1260 Inf.II UPLC with MSDiQ
Several analytical HPLC systems with UV/DAD and/or MS detection with additional capabilities for chiral analysis and fluorescence detection