From Concept to Proof of Concept info@advinus.com   

Absorption, Distribution, Metabolism, Excretion and Toxicity (ADMET)

  • Experienced team with holistic Drug Discovery experience to delineate the druggability issues early in the discovery programs
  • Capability to develop fit for purpose ADME and in vivo pharmacokinetic studies to scientifically support client requirements
  • Expertise in developing quantitative LC-MS/MS methods for complex biomarkers in bio-fluids and tissue samples

Aqueous Solubility

  • Kinetic
  • Thermodynamic
  • Biorelevant media (FaSSIF and FeSSIF)

Lipophilicity (Log D and Log P)

  • Octanol-water/phosphate buffer partitioning

Stability

  • Simulated Gastro Intestinal fluids (SGF, SIF)
  • Biorelevant media (FaSSIF and FeSSIF)
  • Aqueous buffers at different pH
  • Plasma and blood stability
  • Dosing formulation

Pre-formulation

  • Topical formulations
    • Creams/ointment/gel
  • Nanosuspension (bead milling )
  • Microspheres (emulsion)
  1. Metabolic stability/ intrinsic clearance
    • Liver and intestinal microsomes
    • S9 fractions
    • Hepatocytes
    • Direct glucuronidation in liver microsomes using UDPGA
    • Reactive metabolites identification using GSH trapping assay
    • Metabolite finger printing analysis in liver microsomes
  2. Metabolism based drug-drug Interactions
    • CYP inhibition (single point, IC50) using human liver microsomes (HLM)
    • Time dependent CYP inhibition in HLM (IC50 shift, Ki and Kinact)
    • CYP reaction phenotyping (purified rCYPs/microsomes)
    • CYP3A induction assay in HepG2 cell line
    • CYP induction in plateable human hepatocytes
      1. 1A2, 2B6 and 3A4 enzyme activity
      2. mRNA analysis
      3. Cytotoxicity
    • Intrinsic permeability-PAMPA
    • Mono and bi-directional transport: CaCo-2 and MDCK cells
    • In presence and absence of efflux drug transport inhibitors (P-gp, BCRP and MRP)
    • In situ intestinal perfusion with and without mesenteric sampling in rats
    • Definitive plasma protein binding
    • Microsomal  protein binding
    • Brain tissue binding
    • Blood-to-plasma concentration ratio (RBC partitioning)

Cytotoxicity

    • HepG2 cell line
    • Hepatocytes
    • In vitro phototoxicity in 3T3 cell line

Safety

    • 3H radio labeled hERG binding assay

Rodents and non-rodents

    • Rats (multiple strains) : healthy and disease models
    • Mouse (multiple strains) : healthy and disease models
    • Beagle dogs: preferred vendor
    • Various routes of administration
    • Single, multiple and cassette dosing
    • Dose proportionality studies
    • Routes ( urine and feces) of excretion

Tissue exposure kinetics in rodents

    • Exposure kinetics in skin, plasma and other organs  (liver, spleen, thymus etc., ) of choice
    • Partitioning into brain (frontal cortex, striatum, hippocampus and cerebellum)
    • Cerebrospinal fluid collection
    • Partitioning into ocular and related parts (vitreous humor, retina and sclera)-rat

Dermal pharmacokinetics in rodents

    • Healthy and disease models
    • Assistance in selection of  right  animal model

Mechanistic pharmacokinetics models in rodents

  • Chronic bile duct cannulated rats
  • Chronic portal vein cannulated Rats
  • ABT treated mouse and rat pharmacokinetics to delineate involvement of CYP enzymes

PK-PD, modelling and simulations (WinNonlin approach)

    • PK-PD/efficacy correlation – exposure or Cmax based
    • PK modelling  and simulations (fit for purpose)
    • Scaling of  preclinical PK to predict human PK – allometry approach
    • Prediction of starting and efficacious human doses

High-throughput bio-analysis

  • Fit for purpose methods
  • 96 well plate analysis
  • Cassette analysis

Quantitation of biomarkers

  • Quantitative determination for biomarker modulations in plasma and tissues
  • Expertise in quantitation of neurotransmitters and endogenous compounds
  • Expertise in chemical derivatisation techniques to improve chromatographic behaviour and enhance ionization efficiencies

Biotransformation

  • Soft spot analysis to drive structure-activity relationship (SAR) and facilitate in design of metabolically stable analogues
  • Metabolite finger printing analysis
  • Reactive metabolite screening (glutathione conjugation)

Trouble shooting expertise / assessment of matrix effects

  • Evaluation of formulation excipients
  • Evaluation of extraction methods
    • Protein precipitation
    • Liquid-liquid extraction
    • Solid phase extraction
  • Evaluation of ionization polarity   (positive/negative)
  • Evaluation of ionization source (ESI/APCI) technique of analysis (LC-MS/HPLC)

Miscellaneous

LC-MS/MS based in vitro primary and secondary activity screening

  • Biological assays
  • Complex lipid based assays