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EXPERIMENT NO:-01 DATE

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AIM: - To study introduction to in-vitro pharmacology and physiological salt solutions.

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THEORY:-
IN-VITRO PHARMACOLOGY

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What Is In Vitro Pharmacology?
 In In-vitro pharmacology is, in vitro means studies or experiments conducted on

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microorganisms and cells outside of their normal biological environment; and

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pharmacology is the study of the effects of drugs and pharmaceutical products on living
organisms.
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 In pharmacology means research of the biological effects of drugs and pharmaceutical
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products, conducted outside of living organisms.
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 In vitro studies are conducted using components of an organism that have been isolated
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from their usual biological surroundings, such as microorganisms, cells, or biological

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molecules. For example, microorganisms or cells can be studied in artificial culture

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media, and proteins can be examined in solutions.

 Colloquially called "test-tube experiments", these studies in biology, medicine, and their
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sub disciplines are traditionally done in test tubes, flasks, Petri dishes, etc.
In Vitro Pharmacology in Drug Discovery
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 Before drug development comes drug discovery, the process by which new candidate
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drugs are identified.

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 Modern science helps researchers to understand how disease and infections are controlled
at the molecular and physiological levels and to use that knowledge to create compounds
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that will be effective against the chosen targets.

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 The purpose of drug discovery is to identify a drug candidate: a molecule that targets the
chosen disease selectively and effectively, and has favorable medicine-like properties.
 To find such a drug candidate, pharmaceutical researchers screen large libraries of
chemicals for molecules with the necessary properties. Usually, a pool of potentially
useful compounds (“hits”) is identified and narrowed down to identify more promising
compounds (“leads”).

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 The next stage is called lead optimization, where the most promising molecules undergo

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further screening.
 At this stage, in vitro pharmacology assays are used to extensively optimize the

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biological activity and properties of the leads.
 In vitro pharmacology at the lead optimization stage of drug discovery generates high-

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quality data in a precise and timely manner, allowing researchers to identify the best

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compound for their chosen purpose.
 That compound will be the drug candidate that will go on to pre-clinical trials.

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In Vitro Pharmacology in Pre-Clinical Trials

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 When a drug candidate enters the pre-clinical trials phase, researchers already know it is
effective against a certain disease or infection. However, At the pre-clinical trial stage, all
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these parameters must be assessed, and the dose for the first use in a human clinical trial
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must be determined. On average, only one in 5,000 compounds that enter pre-clinical
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trials becomes an approved drug.

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 Its purpose is to assess any potential undesirable effects of the drug on the body’s major
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systems.
At the pre-clinical trial stage, in vitro pharmacology can be used to:
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 Obtain high-quality data on the safety and toxicity of your drug candidate;
 Identify potential adverse effects early in the drug development process;
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 Assess the potency and efficacy of your drug candidate against the targeted disease;
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 Gather data on pharmacokinetics and pharmacodynamics;

 Evaluate the activity of biosimilar compounds.
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Advantages:
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 In vitro pharmacological investigations are significant, in that they permit increasingly

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quick advancement of new medicines - numerous medications can be learned at once and
just those that seem, by all accounts, to be strong go on to human examinations.
Disadvantage:
 A non-attendance of pharmacokinetics, in medicinal phrasing, is one of the critical
downsides of in vitro pharmacological investigations.
 An absence of pharmacokinetics, just as a few different variables, can make it hard to
extrapolate the outcomes to what may be normal when the medication is utilized in vivo.

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PHYSIOLOGICAL SALT SOLUTION (PSS)

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 The ionic requirements and nutritional supply can be provided by using the suitable
solutions, commonly known as PSS.

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 Its composition is such that it provide artificial media resembling the inorganic
composition of blood plasma together with buffer mechanism to maintain the optimum

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pH about 7.0 to 7.2 and glucose to facilitate tissue metabolism.
 Commonly used PSS are frog ringer, Tyrode, De Jalon and Kreb’s solution.

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List and Purpose of each ingredient of various types of PSS :

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Sr. no. Ingredients Concentration in gms/l
Frog
Ringer
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Tyrode Kreb’s De
Jalon
Functions of the
ingredient
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1. Sodium Chloride 6.5 8.0 5.9 9.0 To maintain isotonisity,
(NaCl) isomolarity, contractility
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and excitability
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2. Potassium Chloride 0.14 0.2 0.35 0.350 To provide ionic balance

(KCl)
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3. Calcium Chloride 0.12 0.18 0.28 0.003 To provide contractility

(CaCl2)
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4. Sodium Dihydrogen 0.05 0.1 - - Act as buffer

phosphate (NaH2PO4)
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5. Potassium dihydrogen - - 0.11 - Act as buffer

phosphate (KH2PO4)
6. Magnesium Chloride - 0.1 - - To stabilize the
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(MgCl2) preparation
7. Glucose 1.5 1.0 2.0 0.5 To provide energy
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8. Sodium Bicarbonate 0.4 1.0 2.1 0.5 To provide alkaline

NaHCO3 medium
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QUESTIONS:

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1. What Is In Vitro Pharmacology?
2. Describe used of In Vitro Pharmacology in pre-clinical studies.

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3. What is PSS? Enlist the types PSS solutions.
4. Why sodium bicarbonate is added at the time of setting up of the experiment? Give

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reason.

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5. Which physiological salt solution used for rat/ chicken ileum preparation?
6. What is the function of each composition in Tyrode solution?

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7. What is contractility?

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8. What is Isotonicity? Why there is need to providing isotonic solution?
9. What is the need to provide buffer in PSS?
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