Pharmacokinetics
Studies

Pharmacokinetics Studies

Rodent (different strains of mice and rats) Pharmacokinetics (in –life) studies are conducted in GVK BIO’s vivarium complying with the following - Certification, Compliances and Quality Systems:

Certification and Compliances:

  • AAALAC accredited rodent vivarium facility
  • In house facility for rodent equipped IVC/ Conventional cages
  • OHSAS (Occupational Health & Safety Assessment series) compliant
  • Procedures compliant to Institutional Animal Ethics Committee (IAEC) constituting of External members and Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) nominee
  • Protocols of study experimentation approved by IAEC and reviewed by the committee
  • Institutional Bio Safety Committee (IBSC) registered under Department of Biotechnology, India
  • Bio-waste Disposal Management registered under State Pollution Board
  • EMS (Environment Management System as per ISO 14001)

Quality Systems

  • Procurement of animals from certified vendors: Charles River and Taconic local subsidiaries
  • Documentation of animal health status at receipt 
  • Subjecting all newly arrived animals to quarantine 
  • Physical examination of animals by veterinarian 
  • Subjecting animals for clinical tests:
  • Parasitological examination 
  • Haematological
  • Biochemical 
  • Microbiological examination of faeces for pathogenic organisms on blood agar
  • Feed, water and bedding material analysis
  • Ethical efforts to avoid pain and distress to animals subjected to experimentation
  • Anaesthesia and euthanasia as per CPCSEA guidelines

Ninety SOPs have been established for regular functioning of the vivarium and the various studies being conducted

Cannulations conducted in mice and rats, in house by experienced scientists, to avoid stress to the animals as well as ensure cost-effectiveness

  • Jugular vein 
  • Portal vein 
  • Bile duct 
  • Femoral vein 
  • Double cannulation

Sampling can be done through Portal vein/ tail vein/ saphenous vein/submandibular vein. Wherever possible we employ the principle of 3Rs. In case of mice also, serial sampling is preferred reducing the number of animals and generate quality, robust data.

Routes of Administration

  • Oral
  • Intra-venous 
  • Intra-peritoneal
  • Subcutaneous
  • Intra-muscular
  • Intra-tracheal
  • Sublingual
  • Buccal
  • Intra-duodenal
  • Intracolonic
  • Intranasal
  • Intra-articular

Type of Studies

  • Single/ multiple Doses
  • Dose linearity
  • Tissue Distribution (49 different tissues can be extracted after perfusion): Cold compound
  • Alternate Dosing
  • Cassette Dosing (n=5, inclusive of one reference compound)
  • Broncho-alveolar lavage study
  • Drug-drug Interactions
  • Fed/fasted state
  • Blood Brain Barrier & Cerebrospinal fluid (even from mice)
  • Excretion studies (bile, urine and feces: collected in temperature-maintained metabolic cages)
  • PK/PD
  • Cytochrome P450 Inhibition by 3-Amino-benzotriazole
  • Mechanistic Studies

Strains generally used:

Rats: Wistar, Han Wistar and Sprague Dawley 

Mice: Swiss Albino; CD1; Balb/c; C57BL6

Rabbits, Mini pigs, Guinea pigs

Large Animals: Beagle Dogs (In Collaboration) Large animal PK studies (in life) conducted with external collaborator; which is only Beagle dog breeder in Asia, having a colony of more than 400 dogs at a given time.

Formulation screening is conducted based upon BCS classification and solubility and permeability properties: 

Following steps are employed for preclinical formulation preparation

IV dosing:

  • The solubility of the test compound is first evaluated in phosphate-buffered saline, pH 7.4 (PBS) by visual inspection
  • Other vehicles that are compatible with IV dosing evaluated if the compound is not completely soluble in PBS viz. DMSO, Solutol® HS 15, Captisol and Cremophor EL among others
  • Customized formulation can be accommodated as well. IM, SC, or IN dosing uses the same formulation protocol as IV dosing

PO dosing:

  • The solubility of the test compound is first evaluated in PBS
  • DMSO/ Solutol® HS 15/PBS (5/5/90, v/v/v), or DMSO/1% methylcellulose (5/95, v/v) may be used as the vehicle if the test compound is not completely soluble in PBS at the target concentration
  • Customized formulation can be accommodated as well. IP dosing uses the same formulation protocol as PO dosing

Deliverables as applicable:

  • Plasma or blood concentrations (ng/mL or microM) and tissue concentrations (ng/g tissue) 
  • The ratio of brain concentration to plasma concentration for BBB study
  • Plots of plasma/blood concentration of compound versus time are constructed for PK study
  • Employing protein binding component: unbound, bound and total concentrations can be elucidated
  • Fundamental pharmacokinetic parameters such as AUClast, AUCINF, T½, CL, Vz, Vss, Tmax and Cmax are obtained from the non-compartmental analysis of the plasma/blood data using WinNonlin. Bioavailability (F), if applicable, is also calculated and reported
  • Generally, NCA method is employed, we are also equipped to deliver Compartmental analyses as well as First in Human dose
  • The plasma/blood sample and tissue homogenate (when applicable) preparation method and the HPLC-MS/MS method for each compound are reported
  • Formulation, animal body weight and dosing records are also reported

Typical Compound Requirement for PK study:

Study type TAT (days) Compound Requirement (mg)
Rat PK 5 25
Mouse PK: serial sampling 5 10
Mouse PK: parallel sampling 5 20
Rat BBB * 5 15
Mouse BBB* 5 15
Cassette dosing 5 25 in rats

Assumptions

2 routes: 1 dose: 1 mg/kg iv and 5 mg/kg po, n=3

*1 route:2 routes: 1 dose: 1 mg/kg iv and 5 mg/kg po, n=3

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