IN VITRO


ADME | Drug-Drug Interactions | 3D model | Histopathology | In Vitro Toxicity Research:

In Vitro preclinical toxicology studies. The use of tissue culture provides a valuable solution to address issues of clinical relevance, especially those related to screening, ADME (absorption, distribution, metabolism and excretion), studies of cellular toxicity mechanisms. 3D model in vitro tests for toxicity are beneficial for drug development because of reduced cost, ability to study of mechanism of action, and studies utilizing human cells.

 

We are proud to introduce the in vitro organotypic slice culture (also known as organ culture) model for determination of toxicity of compounds. The advantages of the model is that the system represents

 

Presence of all cell types without any disturbance of natural milieu; Duration-dose dependent changes can be studied (ability to study changes for 14 days or more; not possible with cell lines); Ability to track the cellular changes in real time and derive a dose-response curve.

 

Organ culture is a step closer to use of the actual living animal, as it consists of multicellular tissue supported by stromal elements. At present, we do offer organotypic slice cultures derived from rat pups (post-natal day 7-10) and also from young adult rats. The slices are viable for more than a month following a 7-14 day maturation period. End points to be studied will be custom designed depending on the client requirements.

 

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Most common studies in slice cultures we provide include

  • Viability and Cytotoxicity
  • Cell signaling and communication
  • Invasion and motility
  • Tissue regeneration

 


Quality and accuracy of early in vitro and in vivo studies occupy an essential role in drug discovery and development processes. While the primary goal of ADME (absorption, distribution, metabolism and excretion) study is to discover high-affinity ligands against the target, monitoring of drug-like properties is also accomplished. The foremost reasons for the failure of many new drug candidates are poor Pharmacokinetic (PK), ADME and Toxicological characteristics. The above factors alone account for up to 50% of the attrition rate of new drug candidates during the drug development process. The high cost of R&D and fast pace with which modern medicinal chemists can synthesize new compounds places a high demand on early ADME/PK groups.

 
To optimize the pharmacokinetic characteristics of a drug candidate ADME studies are performed. It helps in identification of the metabolic pathways and provides valuable input for the design of in vivo studies. We offer cost-efficient standardized and customized assays as per the client’s requirement for ADME parameters. ADME capabilities of DRIK include the following:
 

Physicochemical Studies

  • Partition coefficient (LogD/LogP)
  • Solubility (UV/Visible spectroscopy)
  • Chemical stability at different pH
  • Biological matrix stability (serum/plasma/blood/microsomal/hepatocytes/tissue homogenates /simulating fluids)
  • Reversible and time dependent CYP induction
  • 3D Slices

Absorption/Distribution Assays

  • PAMPA (parallel artificial membrane permeability assay)
  • Caco-2 permeability study
  • Tissue permeability studies – Franz Diffusion Assay

Metabolism

  • CYP inhibition/Induction (CYP1A2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4)
  • Drug–Drug Interaction (DDI) studies
  • Pathway determination (Phase I and Phase II)
  • Half-life/clearance determination using Microsomes / Hepatocytes / S9 fractions / CYP across species (Human/Rat/Mouse/Dog)
  • Metabolite identification with Microsomes/Hepatocytes and S9 fraction across species (Rat/Mouse/Dog/Human)

          Note:

          CYP Inhibitor: Furafylline, Pilocarpine, Quercetin, Fluconazole, Ticlopidine,

          CYP Inducer: Omeprazole, Rifampicin, Phenytoin, Quinidine, Ketoconazole

Bio-analysis

We have the capabilities to undertake medium to high throughput Bio-analysis and method development through our partners.

 


Drug screening is a long and costly process. It is big challenge in using animals as model to confirm with low productivity. It limits the discovery of new drugs. To reduce the number of animals in tests and to accelerate drug discovery process, recent efforts have been dedicated to developing cell-based high throughput screening. It helps to improve and fasten the drug screening process. It provides more relevant to in vivo biological information. The data obtained from tissue culture help pharma industry to determine potential efficacy and safety (e.g., predictive toxicity) of drug candidates. In vitro cell culture also provides information about morphology and cellular functions such as cell-cycle status, health and viability, and cell signaling pathways in drug discovery. We provide these services as per the industry requirement.

 

Most common service we provide in the cell lines are

  • Cell viability
  • Cell cytotoxicity
  • Apotosis assay (Caspase 2,3,6,7,8 and 9 assay)
  • Cancer research

 


Stem cells are distinct self-replenishing cell population whose primary function is to generate progeny that develop into terminally differentiated cell types, such as a cardiomyocytes, neurons or receptors. This ability of cells known as “pluripotency” is a hallmark of embryonic stem cells (ESCs). Stem cells belong to one of two major categories according to their potency of differentiation: organ-specific stem cells and pluripotent stem cells. Organ-specific stem cells generally have limited potential for growth and differentiation. In contrast, pluripotent stem cells, such as ESCs and induced pluripotent stem cells (iPSCs) replicate in culture dishes and are theoretically capable of giving rise to any of the cell types found in the body.

 

Most of the drug fails at preclinical stage because of liver toxicity and alterations of hepatic physiology. In addition, drug-induced liver injury is the most common reason for market withdrawal of approved drugs due to safety concerns. It is really difficult to predict liver toxicity because lack of biologically relevant and predictive model systems. Stem cells of hepatocytes will overcome these limitations and provide well-characterized, highly reproducible, and readily available human hepatocytes for preclinical drug development and safety testing.

 

Drik provide services for human induced pluripotent stem cell-derived cardiomyocytes and hepatocytes. These human cardiac and hepatocytes cells are specifically designed to aid drug discovery and improve the predictability against structurally diverse drugs. They help researchers to better understand and optimize the predictive algorithm for applications in discovery toxicology, cardiac and liver safety screening. We obtained these iPSC from reliable source of well-characterized, highly reproducible, and readily available for preclinical drug development and safety testing.

 


To understand mechanism of action, the following assay end points are offered by Drik:

Cell viability

  • MTT
  • Neutral red
  • LDH
  • Cell death
  • Propidium iodide
  • Fluorojade B
  • CYP inhibition; DDI
  • Mitochondrail toxicity
  • Phospholipidosis; Steatosis
  • Apoptosis (Casp3/7/8/9), BrDU
  • Autophagy
  • Necrosis
  • GSH; Oxygen radical absorbance
  • Screening panel


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