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Home Q & A Board Biology-Related Homework Help Cell Biology
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micro micro
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11 years ago
Say a compound has been identified to be antiviral/antimicrobial after determining the IC50, how is it possible to determine the mode of action? e.g. to find out whether the compound changes the membrane structure of the microbe causing the rupture of the cells or damaging/oxidizing the DNA preventing from replication to occurs etc. How about comet assay?
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#1
Valued Member
11 years ago
This is an excellent question. You're asking, how do scientists determine the mechanism of action for new drugs? There are several ways to answer this, because it is not a clear cut procedure. There are a lot of ifs, and depending on where the compound or chemical was found, this could change everything. For example, if the antiviral/antimicrobial is an inorganic chemical (such as an element), it will have a different mode of action than a compound extracted from a live organism. If the compound was extracted from an organism, you'd need to know what elements it is made up of, so you'd take a sample of it and test it use gas chromatography. Gas Chromatography is used to separate volatile compounds in a mixture, so it would tell you the chemical make up of the organic compound. Next, you would chemically probe the compound and feed it to the organism (i.e. human) and see where it concentrates inside the body. If it concentrates in the liver, that could potentially mean that its target location is in the liver. Liver cells, therefore, must contain receptors that bind to the compound. As a result, you'd then isolate liver cells and see what the chemical make-up of the receptors are. Then you can start to piece together how the compound interacts with the receptor. This gets more difficult if we don't know what gene is responsible for creating that receptor (since all receptors are proteins).

Anyway, this would take me a year to answer lol I've attached some valuable resources to help you understand.
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micro Author
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#2
11 years ago
Great reply! What about determining the mode of action in-vitro?
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#3
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11 years ago
Quote from: micro (11 years ago)
What about determining the mode of action in-vitro?

This is all done in vitro:

Quote
As a result, you'd then isolate liver cells and see what the chemical make-up of the receptors are. Then you can start to piece together how the compound interacts with the receptor. This gets more difficult if we don't know what gene is responsible for creating that receptor (since all receptors are proteins).

You mean, what happens after the receptor thing I discussed?
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micro Author
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#4
11 years ago
What I meant is, say you find out that a phytochemical extract (which you determined its composition) is toxic to a virus/bacteria in-vitro; how would you determine if the extract was toxic because it damaged DNA and thus prevented replication or because it disrupted the capsid/membrane bursting the cell etc.
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robertlingrobertling
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11 years ago
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micro Author
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#6
11 years ago
Excellent reply! Cheers buddy. One last question if you don't mind Slight Smile ....so what is meant by "Oxidative damage to DNA in pathology"?
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#7
Valued Member
11 years ago
Thank you. DNA is a very delicate polymer. If its sequence of nucleotides are damaged, the cell could potentially become cancerous because who knows, maybe that nucleotide in that particular gene, was responsible for creating a vital protein for survival. Anyway, cells are constantly producing waste -- essentially they are a micro machine. Some waste products include CO2 and hydrogen peroxide (H2O2). Like the name suggests "Oxidative damage" is when a chemical that contains oxygen, such as hydrogen peroxide, interacts with the nucleotides of the DNA, causing damage to it and consequently wrecks its sequence.
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