DMPK and Metabolism

The study is aimed at evaluation of kinetics and mass balance of absorption of the test item into the test system, distribution among the vital organs/tissues, and biotransformation in the test system and excretion of the test item and its metabolites from the body via exhaled air and excreta (urine, feces and bile).

JRF has extensive experience and expertise to offer ADME to support the clients for Regulatory compliance. The test is prescribed to be undertaken at either for the discovery and developmental phase or re-registration requirements where the product has faced re-investigation.

JRF has highly experienced cross-functional team of scientists involving Toxicologists, Veterinary professionals, Biochemists, and Biostatisticians, for conduct of the experiments, analysis, and interpretation of the experimental data.

The typical DMPK test involves:

• Development of dosing cocktail using 14C&3H tracer-spiked test items (provided by the sponsor).
• Establishing Dose Homogeneity & Stability (LC-MS-MS / LSA)
• Exposure of the mammalian test systems (animals)
• Dosing efficiency
• Collection of the tissues/body fluids, excreta and exhaled gases
• Detection and Quantification of the tracer labeled test item / metabolites in the above
• Total mass balance
• Interpretation of Absorption, Distribution and Excretion Kinetics, including
  • Biliary excretion
  • Protein binding, Blood/plasma partition
  • Metabolite Profiling, Identification, and Quantification
  • Blood Kinetics

1] In vivo Radiolabeled ADME  (Absorption, Distribution, Metabolism & Excretion):

• Animal Models- Rat, Mice
• Radionuclide -14C
• Route of Administration-Oral, Intravenous, Intraperitoneal, Dermal
• Sample Collection and Analysis
  • Specialised glass metabolic cages are used, which facilitate collection of exhaled gases, Urine, Feces, without contact with each other.
  • Blood, Bile, and organs are collected after sacrifice for tracer analysis in the body fluids and tissues.
• Identification and quantification

The studies involve oxidation of the feces and tissues using   Biological Material Oxidiser, followed by the quantification of  the radioactive moieties, by counting the β particles using Liquid Scintillation Counter,  and or HPLC with Flow Scintillation detector, Radio-TLC/Gel plate scanner etc., to achieve mass balance.  Metabolite identification efforts are facilitated by comprehensive interpretations of the analysis conducted using API 4000 Triple Quad Linear Q-Trap LC-MS-MS, coupled with the knowledge of the predictive metabolic pathways. Regulatory Endpoints - Mass Balance, Elimination Kinetics, Route of Elimination, Identification and Quantitation of the metabolite/s.
 

2] Pharmacokinetics / Toxicokinetics / Bioavailability/ Bioequivalence:

• Bioanalysis - Method development and validation using HPLC, HPLC-MS-MS.
• The test substance is detected and quantified for its presence in urine, plasma/serum satisfying USFDA, ICH, EMEA and ENVISA guidance.
• PK/TK Non-compartmental/ Compartmental analysis through WinNonlin software.
• Bioavailability/ Bioequivalence studies in rodents and non-rodents

Services available at following locations:

• JRF India, Gujarat, INDIA, India