UNIT 3 PHARMACEUTICAL MICROBIAL TECHNOLOGY

ANTIMICROBIAL DRUGS
An antimicrobial is an agent that kills microorganisms or stops their growth. Antimicrobial
medicines can be grouped according to the microorganisms they act primarily against. For example,
antibiotics are used against bacteria and antifungals are used against fungi. An antimicrobial is a
substance that kills or inhibits the growth of microorganisms such as bacteria, fungi, or
protozoans. Antimicrobial drugs either kill microbes (microbiocidal) or prevent the growth of
microbes
DISCOVERY OF ANTIMICROBIAL DRUGS
The history of antimicrobials begins with the observations of Pasteur and Koch, who discovered
that one type of bacteria could prevent the growth of another. They did not know at that time
that the reason one bacterium failed to grow was that the other bacterium was producing an
antibiotic. Technically, antibiotics are only those substances that are produced by one
microorganism that kill, or prevent the growth, of another microorganism. In 1928 Alexander
Fleming observed antibiosis against bacteria by a fungus of the genus Penicillium and postulated
the effect was mediated by an antibacterial compound, penicillin, and that its antibacterial
properties could be exploited for chemotherapy. The discovery of antimicrobials like penicillin by
Alexander Fleming and tetracycline paved the way for better health for millions around the
world. Before penicillin became a viable medical treatment in the early 1940s, no true cure for
gonorrhea, strep throat, or pneumonia existed. Patients with infected wounds often had to have a
wounded limb removed, or face death from infection. Now, most of these infections can be cured
easily with a short course of antimicrobials. The term antibiotic was first used in 1942 by
Selman Waksman and his collaborators in journal articles to describe any substance produced by
a microorganism that is antagonistic to the growth of other microorganisms in high dilution. This
definition excluded substances that kill bacteria, but are not produced by microorganisms (such as
gastric juices and hydrogen peroxide). It also excluded synthetic antibacterial compounds such as
the sulfonamides. Many antibacterial compounds are relatively small molecules with a molecular
weight of less than 2000 atomic mass units. With advances in medicinal chemistry, most of
today’s antibacterials chemically are semisynthetic modifications of various natural compounds.
Before the early 20 century, treatments for infections were based primarily on medicinal folklore.
Mixtures with antimicrobial properties that were used in treatments of infections were described
over 2000 years ago. Many ancient cultures, including the ancient Egyptians and ancient Greeks,
used specially selected mold and plant materials and extracts to treat infections. More recent
observations made in the laboratory of antibiosis between micro- organisms led to the discovery
of natural antibacterials produced by microorganisms. Louis Pasteur observed, “if we could
intervene in the antagonism observed between some bacteria, it would offer perhaps the greatest
hopes for therapeutics”. The term ‘antibiosis’, meaning “against life,” was introduced by the
French bacteriologist Vuillemin as a descriptive name of the phenomenon exhibited by these early
antibacterial drugs. Antibiosis was first described in 1877 in bacteria when Louis Pasteur and
Robert Koch observed that an airborne bacillus could inhibit the growth of Bacillus anthracis.
These drugs were later renamed antibiotics by Selman Waksman, an American microbiologist, in
1942.Louis Pasteur was a French microbiologist and chemist best known for their experiments
supporting the Germ theory of disease, and for his vaccinations, most notably the first vaccine
against rabies. John Tyndall first described antagonistic activities by fungi against bacteria in
England in 1875. Synthetic antibiotic chemotherapy as a science and development of antibacterials
began in Germany with Paul Ehrlich in the late 1880s. Ehrlich noted certain dyes would color
human, animal, or bacterial cells, while others did not. He then proposed the idea that it might
be possible to create chemicals that would act as a selective drug that would bind to and kill
bacteria without harming the human host. After screening hundreds of dyes against various
organisms, he discovered a medicinally useful drug, the synthetic antibacterial Salvarsan now
called arsphenamine. In 1895, Vincenzo Tiberio, physician of the University of Naples discovered
that a mold (Penicillium) in a water well has an antibacterial action. After this initial
chemotherapeutic compound proved effective, others pursued similar lines of inquiry, but it was
not until in 1928 that Alexander Fleming observed antibiosis against bacteria by a fungus of the
genus Penicillium. Fleming postulated the effect was mediated by an antibacterial compound
named penicillin, and that its antibacterial properties could be exploited for chemotherapy. He
initially characterized some of its biological properties, but he did not pursue its further
development.
TYPES OF ANTIMICROBIAL DRUGS
Antimicrobial drugs can be divided into two classes based on their mechanism of action.

 Bactericidal drugs kill bacteria;

 Bacteriostatic drugs inhibit their growth or reproduction.
Bactericidal drugs
One way that bactericidal drugs kill bacteria is by inhibiting cell wall synthesis. Examples include the
Beta-lactam antibiotics (penicillin derivatives (penams) ), cephalosporins (cephems), monobactams, and
carbapenems) and vancomycin. Other ways that bactericidal drugs kill bacteria include inhibiting
bacterial enzymes or protein translation. Other batericidal agents include daptomycin, fluoroquinolones,
metronidazole, nitrofurantoin, co-trimoxazole and telithromycin.
Bacteriostatic drugs
Bacteriostatic drugs limit the growth of bacteria by interfering with bacterial protein production, DNA
replication, or other aspects of bacterial cellular metabolism. This group includes: tetracyclines,
sulfonamides, spectinomycin, trimethoprim, chloramphenicol, macrolides and lincosamides. They
must work together with the immune system to remove the microorganisms from the body.
However, there is not always a precise distinction between them and bactericidal antibiotics. High
concentrations of some bacteriostatic agents are also bactericidal.
Antimicrobial drugs are classified further based on their host organisms.

 Antibiotics – drugs against bacteria.

Examples: Penicillin,kanamycin,etc.
 Antifungal – drugs against fungi.
Examples: Amphotericin B, cephalosporins,etc.
 Antiviral – drugs against viruses.
Examples: zanamivir, Ramdesivir,etc.
 Antiprotozoal – drugs against protozoa
Examples: tetracyclin, etc.

Antibiotics.
Antibiotics are a type of antimicrobial medicine that works by suppressing the growth of bacteria and is
used to treat and prevent bacterial illnesses. When used to treat viral infections, such as the common cold
or flu, they are ineffective. Alexander Fleming is credited with the development of antibiotics after
discovering the natural antibiotic penicillin.
Antibiotics function in two ways, despite the fact that there are many different varieties.

1. Certain antibiotics get rid of the bacteria by killing it, like penicillin. They generally disrupt the
formation of the cell wall or cell contents of the bacteria.
2. The other type of bacterium prevents the bacteria from multiplying.

Antifungal drugs. 
Several antifungal antibiotics are currently available for treating infectious disease. One example
is griseofulvin, which is used against the fungi of ringworm and athlete's foot. Other examples
are nystatin, clotrimazole, ketoconazole, and miconazole, all of which are used against vaginal
infections due toCandida albicans. For systemic fungal infections, the antibiotic amphotericin B is
available, although it has serious side effects.
Antiviral drugs. 
Antiviral drugs are not widely available because viruses have few functions or structures with which
drugs can interfere. Nevertheless, certain drugs are available to interfere with viral replication. One
example is azidothymidine (AZT), which is used to interrupt the replication of human
immunodeficiency virus. Other examples are acyclovir, which is used against herpes viruses and
chickenpox viruses;ganciclovir, which is used against cytomega-lovirus; amantadine, which is
prescribed against influenza viruses; and interferon, which has been used against rabies viruses and
certain cancer viruses.
Antiprotozoal drugs. 
Many antibiotics used against bacteria, for example, tetracycline, are also useful against protozoa. Among
the drugs used widely as antiprotozoal agents are metronidazole (Flagyl), which is used
against Trichomonas vaginalis; quinine, which is used against malaria; and pentamidine isethionate,
which is valuable againstPneumocystis carinii.

They are further classified based on their sources as

 Natural – these are naturally produced antimicrobial drugs, mostly derived from other
microorganisms.
Examples: Penicillin, tetracyclins,etc.
 Synthetic – these are chemically synthesized or modified natural antimicrobial drugs.
Examples: Clotrimazole,Sulphanamides.

DRUG RESISTANCE
Antimicrobial resistance is not a new phenomenon. In nature, microbes are constantly evolving in order to
overcome the antimicrobial compounds produced by other microorganisms. Human development of
antimicrobial drugs and their widespread clinical use has simply provided another selective pressure that
promotes further evolution. Several important factors can accelerate the evolution of drug resistance.
These include the overuse and misuse of antimicrobials, inappropriate use of antimicrobials,
subtherapeutic dosing, and patient noncompliance with the recommended course of treatment.
Exposure of a pathogen to an antimicrobial compound can select for chromosomal mutations
conferring resistance, which can be transferred vertically to subsequent microbial generations
and eventually become predominant in a microbial population that is repeatedly exposed to the
antimicrobial. Alternatively, many genes responsible for drug resistance are found
on plasmids or in transposons that can be transferred easily between microbes
through horizontal gene transfer. Transposons also have the ability to move resistance genes
between plasmids and chromosomes to further promote the spread of resistance.

MECHANISMS FOR DRUG RESISTANCE

There are several common mechanisms for drug resistance. These mechanisms include
enzymatic modification of the drug, modification of the antimicrobial target, and prevention of
drug penetration or accumulation.

Drug Modification or Inactivation

Resistance genes may code for enzymes that chemically modify an antimicrobial, thereby
inactivating it, or destroy an antimicrobial through hydrolysis. Resistance to many types of
antimicrobials occurs through this mechanism. For example, aminoglycoside resistance can
occur through enzymatic transfer of chemical groups to the drug molecule, impairing the binding
of the drug to its bacterial target. For β-lactams, bacterial resistance can involve the enzymatic
hydrolysis of the β-lactam bond within the β-lactam ring of the drug molecule. Once the β-
lactam bond is broken, the drug loses its antibacterial activity. This mechanism of resistance is
mediated by β-lactamases, which are the most common mechanism of β-lactam resistance.
Inactivation of rifampin commonly occurs through glycosylation, phosphorylation, or
adenosine diphosphate (ADP) ribosylation, and resistance to macrolides and lincosamides can
also occur due to enzymatic inactivation of the drug or modification.

Prevention of Cellular Uptake or Efflux

Microbes may develop resistance mechanisms that involve inhibiting the accumulation of an
antimicrobial drug, which then prevents the drug from reaching its cellular target. This strategy is
common among gram-negative pathogens and can involve changes in outer membrane lipid
composition, porin channel selectivity, and/or porin channel concentrations. For example, a
common mechanism of carbapenem resistance among Pseudomonas aeruginosa is to decrease
the amount of its OprD porin, which is the primary portal of entry for carbapenems through the
outer membrane of this pathogen. Additionally, many gram-positive and gram-negative
pathogenic bacteria produce efflux pumps that actively transport an antimicrobial drug out of
the cell and prevent the accumulation of drug to a level that would be antibacterial. For example,
resistance to β-lactams, tetracyclines, and fluoroquinolones commonly occurs through active
efflux out of the cell, and it is rather common for a single efflux pump to have the ability to
translocate multiple types of antimicrobials.

Target Modification

Because antimicrobial drugs have very specific targets, structural changes to those targets can
prevent drug binding, rendering the drug ineffective. Through spontaneous mutations in the
genes encoding antibacterial drug targets, bacteria have an evolutionary advantage that allows
them to develop resistance to drugs. This mechanism of resistance development is quite
common. Genetic changes impacting the active site of penicillin-binding proteins (PBPs) can
inhibit the binding of β-lactam drugs and provide resistance to multiple drugs within this class.
This mechanism is very common among strains of Streptococcus pneumoniae, which alter their
own PBPs through genetic mechanisms. In contrast, strains of Staphylococcus aureus develop
resistance to methicillin (MRSA) through the acquisition of a new low-affinity PBP, rather than
structurally alter their existing PBPs. Not only does this new low-affinity PBP provide resistance
to methicillin but it provides resistance to virtually all β-lactam drugs, with the exception of the
newer fifth-generation cephalosporins designed specifically to kill MRSA. Other examples of
this resistance strategy include alterations in

 ribosome subunits, providing resistance to macrolides, tetracyclines, and

aminoglycosides;
 lipopolysaccharide (LPS) structure, providing resistance to polymyxins;
 RNA polymerase, providing resistance to rifampin;
 DNA gyrase, providing resistance to fluoroquinolones;
 metabolic enzymes, providing resistance to sulfa drugs, sulfones,
and trimethoprim; and
 peptidoglycan subunit peptide chains, providing resistance to glycopeptides.

Target Overproduction or Enzymatic Bypass

When an antimicrobial drug functions as an antimetabolite, targeting a specific enzyme to inhibit

its activity, there are additional ways that microbial resistance may occur. First, the microbe may
overproduce the target enzyme such that there is a sufficient amount of antimicrobial-free
enzyme to carry out the proper enzymatic reaction. Second, the bacterial cell may develop a
bypass that circumvents the need for the functional target enzyme. Both of these strategies have
been found as mechanisms of sulfonamide resistance. Vancomycin resistance among S.
aureus has been shown to involve the decreased cross-linkage of peptide chains in the bacterial
cell wall, which provides an increase in targets for vancomycin to bind to in the outer cell wall.
Increased binding of vancomycin in the outer cell wall provides a blockage that prevents free
drug molecules from penetrating to where they can block new cell wall synthesis.

Target Mimicry

A recently discovered mechanism of resistance called target mimicry involves the production of

proteins that bind and sequester drugs, preventing the drugs from binding to their target. For
example, Mycobacterium tuberculosis produces a protein with regular pentapeptide repeats that
appears to mimic the structure of DNA. This protein binds fluoroquinolones, sequestering them
and keeping them from binding to DNA, providing M. tuberculosis resistance to
fluoroquinolones. Proteins that mimic the A-site of the bacterial ribosome have been found to
contribute to aminoglycoside resistance as well.

Multidrug-Resistant Microbes and Cross Resistance

From a clinical perspective, our greatest concerns are multidrug-resistant microbes
(MDRs) and cross resistance. MDRs are colloquially known as “superbugs” and carry one or
more resistance mechanism(s), making them resistant to multiple antimicrobials. In cross-
resistance, a single resistance mechanism confers resistance to multiple antimicrobial drugs. For
example, having an efflux pump that can export multiple antimicrobial drugs is a common way
for microbes to be resistant to multiple drugs by using a single resistance mechanism. Several of
the superbugs discussed in the following sections have been dubbed the ESKAPE pathogens.
This acronym refers to the names of the pathogens (Enterococcus faecium, Staphylococcus
aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas
aeruginosa and Enterobacter spp.) but it is also fitting in that these pathogens are able to
“escape” many conventional forms of antimicrobial therapy. As such, infections by ESKAPE
pathogens can be difficult to treat and they cause a large number of nosocomial infections.