General Characteristics of Antimicrobial Drugs

1
• As Ehrlich so clearly saw, to be successful a chemotherapeutic
agent must have selective toxicity: it must kill or inhibit the
microbial pathogen while damaging the host as little as possible.
• The degree of selective toxicity may be expressed in terms of
• (1) the therapeutic dose, the drug level required for clinical
treatment of a particular infection, and
• (2) the toxic dose, the drug level at which the agent becomes
too toxic for the host.
• The therapeutic index is the ratio of the toxic dose to the
therapeutic dose.
• The larger the therapeutic index, the better the
chemotherapeutic agent (all other things being equal).
• A drug that disrupts a microbial function not found in
eucaryotic animal cells often has a greater selective toxicity and
a higher therapeutic index.
• For example, penicillin inhibits bacterial cell wall peptidoglycan
synthesis but has little effect on host cells because they lack cell
walls; therefore penicillin’s therapeutic index is high.
4
5
• A drug may have a low therapeutic index because it inhibits
the same process in host cells or damages the host in other
ways.
• These undesirable effects on the host, called side effects, are
of many kinds and may involve almost any organ system.
• Because side effects can be severe, chemotherapeutic agents
should be administered with great care.
• Drugs vary considerably in their range of effectiveness.
• Many are narrow-spectrum drugs—that is, they are effective
only against a limited variety of pathogens.
• Others are broad spectrum drugs and attack many different
kinds of pathogens.
• Drugs may also be classified based on the general microbial
group they act against: antibacterial, antifungal, antiprotozoan,
and antiviral.
• Some agents can be used against more than one group; for
example, sulfonamides are active against bacteria and some
protozoa.
• Chemotherapeutic agents can be synthesized by microorganisms
or manufactured by chemical procedures independent of
microbial activity.
• A number of the most commonly employed antibiotics are
natural—that is, totally synthesized by one of a few bacteria or
fungi.
• In contrast, several important chemotherapeutic agents are
completely synthetic.
• Many antiviral and antiprotozoan drugs are synthetic.
• An increasing number of antibiotics are semisynthetic.
• Semisynthetic antibiotics are natural antibiotics that have been
chemically modified by the addition of extra chemical groups
to make them less susceptible to inactivation by pathogens.
• Ampicillin, carbenicillin, and methicillin are good examples.
• Chemotherapeutic agents, like disinfectants, can be either cidal
or static.
• Static agents reversibly inhibit growth; if the agent is removed,
the microorganisms will recover and grow again.
• Although a cidal agent kills the target pathogen, its activity is
concentration dependent and the agent may be only static at
low levels.
• The effect of an agent also varies with the target species: an
agent may be cidal for one species and static for another.
• Because static agents do not directly destroy the pathogen,
elimination of the infection depends on the host’s own
resistance mechanisms.
• A static agent may not be effective if the host’s resistance is too
low.
• Some idea of the effectiveness of a chemotherapeutic agent
against a pathogen can be obtained from the minimal inhibitory
concentration (MIC).
• The MIC is the lowest concentration of a drug that prevents
growth of a particular pathogen.
• The minimal lethal concentration (MLC) is the lowest drug
concentration that kills the pathogen.
• A cidal drug kills pathogens at levels only two to four times the
MIC, whereas a static agent kills at much higher concentration.
13
 Determining the Level of Antimicrobial Activity

• Determination of antimicrobial effectiveness against specific

pathogens is essential to proper therapy.
• Testing can show which agents are most effective against a
pathogen and give an estimate of the proper therapeutic dose.
 Dilution Susceptibility Tests

• Dilution susceptibility tests can be used to determine MIC

and MLC values.
• In the broth dilution test, a series of broth tubes (usually
Mueller-Hinton broth) containing antibiotic concentrations in
the range of 0.1 to 128 g/ml is prepared and inoculated with
standard numbers of the test organism.
• The lowest concentration of the antibiotic resulting in no
growth after 16 to 20 hours of incubation is the MIC.
• The MLC can be ascertained if the tubes showing no
growth are subcultured into fresh medium lacking
antibiotic.
• The lowest antibiotic concentration from which the
microorganisms do not recover and grow when
transferred to fresh medium is the MLC.
• The agar dilution test is very similar to the broth dilution
test.
• Plates containing Mueller-Hinton agar and various amounts
of antibiotic are inoculated and examined for growth.
• Recently several automated systems for susceptibility testing
and MIC determination with broth or agar cultures have been
developed.

17
 Disk Diffusion Tests

• If a rapidly growing aerobic or facultative pathogen like

Staphylococcus or Pseudomonas is being tested, a disk
diffusion technique may be used to save time and media.
• The principle behind the assay technique is fairly simple.
• When an antibiotic-impregnated disk is placed on agar
previously inoculated with the test bacterium, the disk picks
up moisture and the antibiotic diffuses radially outward
through the agar, producing an antibiotic concentration
gradient.
• The antibiotic is present at high concentrations near the disk
and affects even minimally susceptible microorganisms
resistant organisms will grow up to the disk).
• As the distance from the disk increases, the antibiotic
concentration drops and only more susceptible
pathogens are harmed.
• A clear zone or ring is present around an antibiotic disk
after incubation if the agent inhibits bacterial growth.
• The wider the zone surrounding a disk, the more
susceptible the pathogen is. Zone width also is a
function of the antibiotic’s initial concentration, its
solubility, and its diffusion rate through agar.
• Thus zone width cannot be used to compare directly the
effectiveness of two different antibiotics.
• Currently the disk diffusion test most often used is the
Kirby-Bauer method, which was developed in the early
1960s at the University of Washington Medical School by
William Kirby, A.W. Bauer, and their colleagues.
• An inoculating loop or needle is touched to four or five
isolated colonies of the pathogen growing on agar and then
used to inoculate a tube of culture broth.
• The culture is incubated for a few hours at 35°C until it
becomes slightly turbid and is diluted to match a turbidity
standard.
• A sterile cotton swab is dipped into the standardized bacterial
test suspension and used to evenly inoculate the entire surface
of a Mueller-Hinton agar plate.
• After the agar surface has dried for about 5 minutes, the
appropriate antibiotic test disks are placed on it, either with
sterilized forceps or with a multiple applicator device.
• The plate is immediately placed in a 35°C incubator.
• After 16 to 18 hours of incubation, the diameters of the zones
of inhibition are measured to the nearest mm.
• Kirby-Bauer test results are interpreted using a table that
relates zone diameter to the degree of microbial resistance.
• A plot of MIC (on a logarithmic scale) versus zone inhibition
diameter (arithmetic scale) is prepared for each antibiotic.
• These plots are then used to find the zone diameters
corresponding to the drug concentrations actually reached in
the body.
• If the zone diameter for the lowest level reached in the body
is smaller than that seen with the test pathogen, the
pathogen should have an MIC value low enough to be
destroyed by the drug.
• A pathogen with too high an MIC value (too small a zone
diameter) is resistant to the agent at normal body
concentrations.
 Measurement of Drug Concentrations in the Blood

• A drug must reach a concentration at the site of infection

above the pathogen’s MIC to be effective.
• In cases of severe, life threatening disease, it often is
necessary to monitor the concentration of drugs in the blood
and other body fluids.
• This may be achieved by microbiological, chemical,
immunologic, enzymatic, or chromatographic assays.
Antibacterial Drugs
 Sulfonamides or Sulfa Drugs
• A good way to inhibit or kill pathogens is by use of
compounds that are structural analogues, molecules
structurally similar to metabolic intermediates.
• These analogues compete with metabolites in metabolic
processes because of their similarity, but are just different
enough so that they cannot function normally in cellular
metabolism.
• The first antimetabolites to be used successfully as
chemotherapeutic agents were the sulfonamides, discovered
by G. Domagk.
• Sulfonamides or sulfa drugs are structurally related to
sulfanilamide, an analogue of p-aminobenzoic acid.
• The latter substance is used in the synthesis of the cofactor
folic acid.
• sulfanilamide or another sulfonamide enters a bacterial cell, it
competes with p-aminobenzoic acid for the active site of an
enzyme involved in folic acid synthesis, and the folate
concentration decreases.
• The decline in folic acid is detrimental to the bacterium
because folic acid is essential to the synthesis of purines and
pyrimidines, the bases used in the construction of DNA, RNA,
and other important cell constituents.
• The resulting inhibition of purine and pyrimidine synthesis leads to
cessation of bacterial growth or death of the pathogen.
• Sulfonamides are selectively toxic for many pathogens because
these bacteria manufacture their own folate and cannot effectively
take up the cofactor.
• In contrast, humans cannot synthesize folate and must obtain it in
the diet; therefore sulphonamides will not affect the host.
• The effectiveness of sulfonamides is limited by the increasing
sulfonamide resistance of many bacteria.
• Furthermore, as many as 5% of the patients receiving sulfa drugs
experience adverse side effects, chiefly in the form of allergic
responses (fever, hives, and rashes).
 Quinolones

• A second group of synthetic antimicrobial agents are

increasingly used to treat a wide variety of infections.
• The quinolones are synthetic drugs that contain the 4-
quinolone ring.
• The first quinolone, nalidixic acid, was synthesized in 1962.
• Three of these—ciprofloxacin, norfloxacin, and ofloxacin—are
currently used in the United States, and more fluoroquinolones
are being synthesized and tested.
• Quinolones are effective when administered orally.
• They sometimes cause adverse side effects, particularly
gastrointestinal upset.
• Quinolones act by inhibiting the bacterial DNA gyrase or
topoisomerase II, probably by hindering to the DNA gyrase
complex.
• This enzyme introduces negative twists in DNA and helps
separate its strands.
• DNA gyrase inhibition disrupts DNA replication and repair,
transcription, bacterial chromosome separation during division,
and other cell processes involving DNA.
• Fluoroquinolones also inhibit topoisomerase IV, another
enzyme that untangles DNA during replication.
• The quinolones are broad-spectrum drugs. They are highly
effective against enteric bacteria such as E. coli and Klebsiella
pneumoniae.
• They can be used with Haemophilus, Neisseria, Pseudomonas
aeruginosa, and other gram-negative pathogens.
• The quinolones also are active against gram-positive bacteria
such as Staphylococcus aureus, Streptococcus pyogenes, and
Mycobacterium tuberculosis.
• Currently they are used in treating urinary tract infections,
sexually transmitted diseases caused by Neisseria and
Chlamydia, gastrointestinal infections, respiratory tract
infections, skin infections, and osteomyelitis.
 Penicillins
• Penicillin G or benzylpenicillin, the first antibiotic to be widely used
in medicine, has the structural properties characteristic of the
penicillin family.
• Most penicillins are derivatives of 6-aminopenicillanic acid and
differ from one another only with respect to the side chain attached
to its amino group.
• The most crucial feature of the molecule is the β-lactam ring, which
appears to be essential for activity.
• Penicillinase, the enzyme synthesized by many penicillin-resistant
bacteria, destroys penicillin activity by hydrolyzing a bond in this
ring.
• Their structures do resemble that of the terminal D-alanyl-D-
alanine found on the peptide side chain of the peptidoglycan
subunit.
• It has been proposed that penicillins inhibit the enzyme
catalyzing the transpeptidation reaction because of their
structural similarity, which would block the synthesis of a
complete, fully cross-linked peptidoglycan and lead to osmotic
lysis.
• The mechanism is consistent with the observation that
penicillins act only on growing bacteria that are synthesizing
new peptidoglycan.
• However, more recently it has been discovered that penicillins
bind to several penicillin-binding proteins and may destroy
bacteria by activating their own autolytic enzymes.
• There now is some evidence that penicillin kills bacteria even in
the absence of autolysins or murein hydrolases.
• Lysis could occur after bacterial viability has already been lost.
• Penicillin may stimulate special proteins called bacterial holins
to form holes or lesions in the plasma membrane.
• This would directly lead to membrane leakage and death;
murein hydrolases also could move through the holes, disrupt
the peptidoglycan, and lyse the cell.
• Penicillins differ from each other in several ways.
• Penicillin G is effective against gonococci, meningococci, and
several grampositive pathogens such as streptococci and
staphylococci, but it must be administered parenterally
because it is destroyed by stomach acid.
• Penicillin V is similar to penicillin G, but it is more acid resistant
and can be given orally.
• Ampicillin can be administered orally and has a broader
spectrum of activity as it is effectiveagainst gram-negative
bacteria such as Haemophilus, Salmonella, and Shigella.
• Carbenicillin and ticarcillin also are broad spectrum and
particularly potent against Pseudomonas and Proteus.
• An increasing number of bacteria are penicillin resistant.
• Penicillinase-resistant penicillins such as methicillin, nafcillin,
and oxacillin are frequently employed against these bacterial
pathogens.
• Although penicillins are the least toxic of the antibiotics, about
1 to 5% of the adults in the United States are allergic to them.
 Cephalosporins

• Cephalosporins are a family of antibiotics originally isolated in

1948 from the fungus Cephalosporium, and their β-lactam
structure is very similar to that of the penicillins.
• As might be expected from their structural similarities,
cephalosporins resemble penicillins in inhibiting the
transpeptidation reaction during peptidoglycan synthesis.
• They are broad-spectrum drugs frequently given to patients
with penicillin allergies.
• Many cephalosporins are in use. There are three groups or
generations of these drugs that differ in their spectrum of
activity.
• First-generation cephalosporins are more effective against
gram-positive than gram-negative pathogens.
• Second generation drugs act against many gram-negative as
well as grampositive pathogens.
• Third-generation drugs are particularly effective against gram-
negative pathogens, and often also reach the central nervous
system.
• Most cephalosporins (including cephalothin, cefoxitin,
ceftriaxone, and cefoperazone) are administered parenterally.
• Cefoperazone is resistant to destruction by β- lactamases and
effective against many gram-negative bacteria, including
Pseudomonas aeruginosa.
• Cephalexine and cefixime are given orally rather than by
injection.
 The Tetracyclines

• The tetracyclines are a family of antibiotics with a common four-

ring structure to which a variety of side chains are attached.
• Oxytetracycline and chlortetracycline are naturally produced by
some species of the actinomycete genus Streptomyces; others
are semisynthetic drugs.
• These antibiotics inhibit protein synthesis by combining with the
small (30S) subunit of the ribosome and inhibiting the binding of
aminoacyltRNA molecules to the ribosomal A site.
• Because their action is only bacteriostatic, the effectiveness of
treatment depends on active host resistance to the pathogen.
• Tetracyclines are broad-spectrum antibiotics active against gram-
negative bacteria, gram-positive bacteria, rickettsias,
chlamydiae, and mycoplasmas.
• High doses may result in nausea, diarrhea, yellowing of teeth in
children, and damage to the liver and kidneys.
 Aminoglycoside Antibiotics
• There are several important aminoglycoside antibiotics.
• Streptomycin, kanamycin, neomycin, and tobramycin are
synthesized by Streptomyces, whereas gentamicin comes from a
related bacterium, Micromonospora purpurea.
• Although there is considerablevariation in structure among the
different aminoglycosides, all contain a cyclohexane ring and
amino sugars.
• Aminoglycosides bind to the small ribosomal subunit and
interfere with protein synthesis in at least two ways.
• They directly inhibit protein synthesis and also cause
misreading of the genetic message carried by mRNA.
• The aminoglycosides are bactericidal and tend to be most
active against gram-negative pathogens.
• Streptomycin’s usefulness has decreased greatly due to
widespread drug resistance, but it is still effective against
tuberculosis and plague.
• Gentamicin is used to treat Proteus, Escherichia, Klebsiella,
and Serratia infections.
• Aminoglycosides are quite toxic and can cause deafness, renal
damage, loss of balance, nausea, and allergic responses
 Erythromycin and Other Macrolides
• Erythromycin, the most frequently used macrolide antibiotic,
is synthesized by Streptomyces erythraeus.
• The macrolides contain a 12- to 22-carbon lactone ring linked
to one or more sugars.
• Erythromycin is usually bacteriostatic and binds with the 23S
rRNA of the 50S ribosomal subunit to inhibit peptide chain
elongation during protein synthesis.
• Erythromycin is a relatively broad-spectrum antibiotic
effective against gram-positive bacteria, mycoplasmas, and a
few gram-negative bacteria.
• It is used with patients allergic to penicillins and in the
treatment of whooping cough, diphtheria, diarrhea
caused by Campylobacter, and pneumonia from
Legionella or Mycoplasma infections.
• Newer macrolides are now in use.
• Clindamycin is effective against a variety of bacteria
including staphylococci and anaerobes such as
Bacteroides.
• Azithromycin is particularly effective against Chlamydia
trachomatis.
 Vancomycin and Teicoplanin
• Vancomycin is a glycopeptide antibiotic produced by
Streptomyces orientalis.
• It is a cup-shaped molecule composed of a peptide linked to a
disaccharide.
• The antibiotic blocks peptidoglycan synthesis by inhibiting the
transpeptidation step that cross-links adjacent peptidoglycan
strands.
• The resulting peptidoglycan is mechanically weak and the cells
osmotically lyse.
• Vancomycin’s peptide portion binds specifically to the D-
alanine-D-alanine terminal sequence on the pentapeptide
portion of peptidoglycan.
• This complex blocks transpeptidase action.
• The antibiotic is bactericidal for Staphylococcus and some
members of the genera Clostridium, Bacillus, Streptococcus,
and Enterococcus.
• It is given both orally and intravenously, and has been
particularly important in the treatment of antibiotic resistant
staphylococcal and enterococcal infections.
• Vancomycin-resistant strains of Enterococcus have become
widespread and recently a few cases of resistant Staphylococcus
aureus have appeared.
• Teicoplanin is a glycopeptide antibiotic from Actinoplanes
teichomyceticus that is similar in structure and mechanism of
action to vancomycin.
• It is active against staphylococci, enterococci, streptococci,
clostridia, Listeria, and many other grampositive pathogens.
 Chloramphenicol
• Although chloramphenicol was first produced from cultures of
Streptomyces venezuelae, it is now made through chemical
synthesis.
• Like erythromycin, chloramphenicol binds to 23S rRNA on the
50S ribosomal subunit.
• It inhibits the peptidyl transferase and is bacteriostatic.
• This antibiotic has a very broad spectrum of activity but
unfortunately is quite toxic.
• One may see allergic responses or neurotoxic reactions.
• The most common side effect is a temporary or permanent
depression of bone marrow function, leading to aplastic anemia
and a decreased number of blood leukocytes.
Antifungal Drugs
• Treatment of fungal infections generally has been less
successful than that of bacterial infections largely
because Eucaryotic fungal cells are much more similar to
human cells than are bacteria.
• Many drugs that inhibit or kill fungi are therefore quite
toxic for humans.
• In addition, most fungi have a detoxification system that
modifies many antibiotics, probably by hydroxylation.
• Despite their relatively low therapeutic index, a few drugs
are useful intreating many major fungal diseases.
• Effective antifungal agents frequently either extract
membrane sterols or prevent their synthesis.
• Similarly, because animal cells do not have cell walls, the
enzyme chitin synthase is the target for fungal-active
antibiotics such as polyoxin D and nikkomycin.
• Fungal infections are often subdivided into infections of
superficialtissues or superficial mycoses and systemic
mycoses.
• Treatment for these two types of disease is very different.
Several drugs are used to treat superficial mycoses.
• Three drugs containing imidazole—miconazole,
ketoconazole, and clotrimazole—are broad-spectrum
agents available as creams and solutions for the
treatment of dermatophyte infections such as athlete’s
foot, and oral and vaginal candidiasis .
• They are thought to disrupt fungal membrane permeability
and inhibit sterol synthesis.
• Tolnaftate is used topically for the treatment of cutaneous
infections, but is not as effective against infections of the
skin and hair.
• Nystatin, a polyene antibiotic from Streptomyces, is used to
control Candida infections of the skin, vagina, or alimentary
tract.

• Griseofulvin, an antibiotic formed by Penicillium, is given

orally to treat chronic dermatophyte infections.
• It is thought to disrupt the mitotic spindle and inhibit cell
division; it also may inhibit protein and nucleic acid
synthesis.
• Side effects of griseofulvin include headaches,
gastrointestinal upset and allergic reactions.
• Superficial and systemic mycoses Systemic infections are
very difficult to control and can be fatal.
• Three drugs commonly used against systemic mycoses
are amphotericin B, 5-flucytosine, and fluconazole.
• Amphotericin B from Streptomyces spp. binds to the
sterols in fungal membranes, disrupting membrane
permeability and causing leakage of cell constituents.
• It is quite toxic and used only for serious, life-threatening
infections.
• The synthetic oral antimycotic agent 5-flucytosine (5-
fluorocytosine) is effective against most systemic fungi,
although drug resistance often develops rapidly.
• The drug is converted to 5-fluorouracil by the fungi,
incorporated into RNA in place of uracil, and disrupts
RNA function.
• Its side effects include skin rashes, diarrhea, nausea,
aplastic anemia, and liver damage.
• Fluconazole is used in the treatment of candidiasis,
cryptococcal meningitis, and coccidioidal meningitis.
• Adverse effects are relatively uncommon.
• Just as with antibacterial drugs, overuse of
antifungal agents leads to an increase in drug
resistance.
• For example, Candida infections are becoming
more frequent and drug resistant.
Antiviral Drugs
• For many years the possibility of treating viral infections
with drugs appeared remote because viruses enter host
cells and make use of host cell enzymes and constituents
to a large extent.
• A drug that would block virus reproduction also was
thought to be toxic for the host.
• Inhibitors of virus-specific enzymes and life cycle
processes have now been discovered, and several drugs
are used therapeutically.
• Most antiviral drugs disrupt either critical stages in the virus
life cycle or the synthesis of virus-specific nucleic acids.
• Amantadine and rimantadine can be used to prevent
influenza A infections.
• When given in time, it will reduce the incidence of influenza
by 50 to 70% in an exposed population.
• Amantadine blocks the penetration and uncoating of
influenza virus particles.

73
• Adenine arabinoside or vidarabine disrupts the activity of
DNA polymerase and several other enzymes involved in
DNA and RNA synthesis and function.
• It is given intravenously or applied as an ointment to treat
herpes infections.
• A third drug, acyclovir, is also used in the treatment of
herpes infections.
• Upon phosphorylation, acyclovir resembles deoxy-GTP
and inhibits the virus DNA polymerase.
• Unfortunately acyclovirresistant strains of herpes are
already developing.
• Effective acyclovir derivatives and relatives are now
available.
• Valacyclovir is an orally administered prodrug form of
acyclovir.
• Ganciclovir, penciclovir, and its oral form famciclovir are
effective in treatment of herpesviruses.
• Another kind of drug, foscarnet, inhibits the virus DNA
polymerase in a different way.
• Foscarnet is an organic analogue of pyrophosphate that
binds to the polymerase active site and blocks the
cleavage of pyrophosphate from nucleoside triphosphate
substrates.
• It is used in treating herpes and cytomegalovirus
infections.
• Several broad-spectrum anti-DNA virus drugs have been
developed.
• A good example is the drug HPMPC or cidofovir.
• It is effective against papovaviruses, adenoviruses,
herpesviruses, iridoviruses, and poxviruses.
• The drug acts on the viral DNA polymerase as a
competitive inhibitor and alternative substrate of dCTP.
• It has been used primarily against cytomegalovirus but
also against herpes simplex and human papillomavirus
infections Research on anti-HIV drugs has been
particularly active.
• Many of the first drugs to be developed were reverse
transcriptase inhibitors such as azidothymidine (AZT) or
zidovudine, lamivudine (3TC), didanosine (ddI),
zalcitabine (ddC), and stavudine (d4T).
• These interfere with reverse transcriptase activity and
therefore block HIV reproduction.
• More recently HIV protease inhibitors have been
developed.
• Three of the most used are saquinvir, indinavir, and
ritonavir.
• These mimic the peptide bond that is normally attacked by
the protease.
• The most successful treatment regimen involves a cocktail
of agents given at high dosages to prevent the development
of drug resistance.
• For example, the combination of AZT, 3TC, and ritonavir is
very effective in reducing HIV plasma concentrations almost
to zero.
• However, the treatment does not seem able to eliminate
latent proviral HIV DNA that still resides in memory T cells,
and possibly elsewhere.
• Probably the most publicized antiviral agents are
interferons.
• These small proteins, produced by the host, inhibit virus
replication and may be clinically useful in the treatment of
influenza, hepatitis, herpes, and colds.
 Mechanisms of Action of
antibiotics/Antimicrobial Agents
Antimicrobial Agents

85
86
87
• It is important to know something about the
mechanisms of drug action because such
knowledge helps explain the nature and
degree of selective toxicity of individual drugs
and sometimes aids in the design of new
chemotherapeutic agents.
• The most selective antibiotics are those that interfere with the
synthesis of bacterial cell walls (e.g., penicillins, cephalosporins,
vancomycin, and bacitracin).
• These drugs have a high therapeutic index because bacterial
cell walls have a unique structure not found in eucaryotic cells.
• Streptomycin, gentamicin, spectinomycin, clindamycin,
chloramphenicol, tetracyclines, erythromycin, and many other
antibiotics inhibit protein synthesis by binding with the
prokaryotic ribosome.
• Because these drugs discriminate between prokaryotic and
eucaryotic ribosomes, their therapeutic index is fairly high,
but not as favorable as that of cell wall synthesis inhibitors.
• Some drugs bind to the 30S (small) subunit, while others
attach to the 50S (large) ribosomal subunit.
• Several different steps in the protein synthesis mechanism can
be affected: aminoacyl-tRNA binding, peptide bond
formation, mRNA reading, and translocation.
• For example, fusidic acid binds to EF-G and blocks
translocation, whereas mucopirocin inhibits isoleucyl-tRNA
synthetase.
• The antibacterial drugs that inhibit nucleic acid synthesis or
damage cell membranes often are not as selectively toxic as
other antibiotics.
• This is because procaryotes and eucaryotes do not differ greatly
with respect to nucleic acid synthetic mechanisms or cell
membrane structure.
• Good examples of drugs that affect nucleic acid synthesis or
membrane structure are quinolones and polymyxins.
• Quinolones inhibit the DNA gyrase and thus interfere with DNA
replication, repair, and transcription.
• Polymyxins act as detergents or surfactants and disrupt the
bacterial plasma membrane.
• Several valuable drugs act as antimetabolites: they block the
functioning of metabolic pathways by competitively inhibiting the
use of metabolites by key enzymes.
• Sulfonamides and several other drugs inhibit folic acid
metabolism. Sulfonamides (e.g., sulfanilamide, sulfamethoxazole,
and sulfacetamide) have a high therapeutic index because
humans cannot synthesize folic acid and must obtain it in their
diet.
• Most bacterial pathogens synthesize their own folic acid and are
therefore susceptible to inhibitors of folate metabolism.
• Antimetabolite drugs also can inhibit other pathways.
• For example, isoniazid interferes with either pyridoxal or NAD
metabolism.
 Factors Influencing the Effectiveness of
Antimicrobial Drugs

• It is crucial to recognize that drug therapy is not a simple

matter.
• Drugs may be administered in several different ways, and they
do not always spread rapidly throughout the body or
immediately kill all invading pathogens.
• A complex array of factors influence the effectiveness of drugs.
• First, the drug must actually be able to reach the site of
infection.
• The mode of administration plays an important role.
• A drug such as penicillin G is not suitable for oral
administration because it is relatively unstable in stomach
acid.
• Some antibiotics—for example, gentamicin and other
aminoglycosides—are not well absorbed from the intestinal
tract and must be injected intramuscularly or given
intravenously.
• Other antibiotics (neomycin, bacitracin) are applied topically
to skin lesions.
• Non oral routes of administration often are called parenteral
routes.
• Even when an agent is administered properly, it may be
excluded from the site of infection.
• For example, blood clots or necrotic tissue can protect
bacteria from a drug, either because body fluids containing
the agent may not easily reach the pathogens or because the
agent is absorbed by materials surrounding it.
• Second, the pathogen must be susceptible to the drug.
• Bacteria in abscesses may be dormant and therefore resistant
to chemotherapy, because penicillins and many other agents
affect pathogens only if they are actively growing and
dividing.
• A pathogen, even though growing, may simply not be
susceptible to a particular agent.
• For example, penicillins and cephalosporins, which inhibit cell
wall synthesis, do not harm mycoplasmas, which lack cell
walls.
• Third, the chemotherapeutic agent must exceed the
pathogen’s MIC value if it is going to be effective.
• The concentration reached will depend on the amount of drug
administered, the route of administration and speed of
uptake, and the rate at which the drug is cleared or eliminated
from the body.
• It makes sense that a drug will remain at high concentrations
longer if it is absorbed over a long period and excreted slowly.
• Finally, chemotherapy has been rendered a less effective and
much more complex matter by the spread of drug resistance
plasmids.
 Resistance Factors

• Plasmids often confer antibiotic resistance on the bacteria

that contain them.
• R factors or plasmids typically have genes that code for
enzymes capable of destroying or modifying antibiotics.
• They are not usually integrated into the host chromosome.
• Genes coding for resistance to antibiotics such as ampicillin,
chloramphenicol, and kanamycin have been found in
plasmids.
• Some R plasmids have only a single resistance gene, whereas
others can have as many as eight.
• Often the resistance genes are within a transposon, and thus
it is possible for bacterial strains to rapidly develop multiple
resistance plasmids.
• Because many R factors also are conjugative plasmids, they
can spread throughout a population, although not as rapidly
as the F factor.
• Often, non conjugative R factors also move between bacteria
during plasmid promoted conjugation. Thus a whole
population can become resistant to antibiotics.
• The fact that some of these plasmids are readily
transferred between species further promotes the
spread of resistance.
• When the host consumes large quantities of antibiotics,
E. coli and other bacteria with R factors are selected for
and become more prevalent.
• The R factors can then be transferred to more
pathogenic genera such as Salmonella or Shigella,
causing even greater public health problems.
 Col Plasmids

• Bacteria also harbor plasmids with genes that may give

them a competitive advantage in the microbial world.
• Bacteriocins are bacterial proteins that destroy other
bacteria.
• They usually act only against closely related strains.
• Bacteriocins often kill cells by forming channels in the
plasma membrane, thus increasing its permeability.
• They also may degrade DNA and RNA or attack
peptidoglycan and weaken the cell wall.
• Col plasmids contain genes for the synthesis of
bacteriocins known as colicins, which are directed against
E. coli. Similar plasmids carry genes for bacteriocins
against other species.
• For example, Col plasmids produce cloacins that kill
Enterobacter species.
• Clearly the host is unaffected by the bacteriocin it
produces.
• Some Col plasmids are conjugative and also can carry
resistance genes.
 Other Types of Plasmids

• Several other important types of plasmids have been

discovered.
• Some plasmids, called virulence plasmids, make their
hosts more pathogenic because the bacterium is better
able to resist host defense or to produce toxins.
• For example, enterotoxigenic strains of E. coli cause
traveler’s diarrhea because of a plasmid that codes for an
enterotoxin.
• Metabolic plasmids carry genes for enzymes that
degrade substances such as aromatic compounds
(toluene), pesticides (2,4-dichlorophenoxyacetic acid),
and sugars (lactose).
• Metabolic plasmids even carry the genes required for
some strains of Rhizobium to induce legume
nodulation and carry out nitrogen fixation.