Chapter 20

Antimicrobial Drugs
 Antimicrobial Drugs

• Chemotherapy
– The use of drugs to treat a disease
• Antimicrobial drugs
– Interfere with the growth of microbes within a host
– Must exhibit selective toxicity
• Selective toxicity
– A drug that kills harmful microbes without damaging
the host
• Antibiotic
– A substance produced by a microbe that, in small
amounts, inhibits another microbe

 Antimicrobial
Drugs
• Fleming discovered
penicillin, produced
by Penicillium
– S. aureus growth
inhibited by mold
• Many species of soil
bacteria are known
inhibit growth of
other bacteria
– Streptomyces

Sources of Antibiotics
 The Spectrum of
Antimicrobial Activity

• Human Bacterial Infections

– Easier to treat because bacteria are prokaryotic (and
humans are eukaryotic)
– Fungi and protozoa more difficult (similar cells)
– Virus most difficult (using host cell to make viruses)
• Broad spectrum
– Treat many types of organisms
• Narrow spectrum
– Treat few types of microorganisms
• Superinfection
– Pathogen that develops antibiotic resistance

Examples of Effective Antibiotics

 The Action of
Antimicrobial Drugs

• Bactericidal
– Kill microbes directly

• Bacteriostatic
– Prevent microbes from growing

The Action of Antimicrobial Drugs

 Cell Wall Synthesis Inhibitors

• Cell Wall Rupture

– One mechanism for some antibiotics
• Penicillin
– Incorporated into cell wall
• Interferes with peptidoglycan synthesis and assembly
– Cell wall becomes weak and unstable
– Bacterium ruptures and dies
– Only effective if bacteria are multiplying
• i.e. manufacturing cell walls
• Slow growing bacteria are problematic
• Why don’t antibiotics lyse human cells?
– (Hint: What do bacteria have that human cells don’t?)

Cell Wall Damage

Cell Wall Synthesis Inhibitors: Natural Penicillins
• Extracted from cultures of Penicillium
• Penicillin
– Good for use on staphylococci, streptococci,
spirochaetes
– Narrow spectrum
– Susceptible to penicillinase (many bacteria produce)
• Also called β-lactamase (because penicillin contains a β-lactam
ring)
• Penicillin G
– Base compound
– Diminished activity if taken orally
– Excreted in 3-6 hours
• Penicillin V
– Stable in stomach acid

Cell Wall Synthesis Inhibitors: Other Penicillins

• Semisynthetic penicillins
– Natural penicillin is collected
– Modified chemically to achieve better characteristics
• Penicillinase Resistant Penicillins
– Methicillin
– Oxacillin
• Extended-spectrum penicillins
– Wider range of target species (developed in lab)
– Aminopenicillins
• Ampicillin, amoxicillin (until resistance developed then...)
– Carboxypenicillins
• Carbenicillin, and ticarcillin
– Ureidopenicillins (newest)
• Mezlocillin and azlocillin

The Structure of Penicillins

The Structure of Penicillins
The Effect of Penicillinase on Penicillins
Other Notable Cell Wall Inhibitors
• Cephalosporins
• Polypeptide antibiotics
– Bacitracin
• Derived from a Bacillus
• Topical application against gram-positives
– Vancomycin
• Derived from Streptomyces
• Very narrow spectrum
• Important "last line" against antibiotic-resistant
S. aureus (MRSA)
• Antimycobacterial antibiotics
– Isoniazid
• Inhibits mycolic acid synthesis
• Good to treat Mycobacterium tuberculosis (slow growers)
– Ethambutol
• Inhibits incorporation of mycolic acid

Protein Synthesis Inhibition by Antibiotics

 Inhibitors of Protein Synthesis
• Chloramphenicol
– Broad spectrum
• Binds 50S subunit; inhibits peptide bond formation
– Inexpensive to manufacture
• Low cost to consumer
– Suppresses bone marrow activity
• Sometimes causes severe anemia

 Inhibitors of Protein Synthesis

• Aminoglycosides
– Streptomycin, neomycin, gentamicin
• Broad spectrum
– Changes shape of 30S subunit
– First antibiotics to affect gram-negatives
– Possible side effects
• Permanent damage to auditory nerve
• Kidney damage
– Gentamicin
• Particularly good against Pseudomonas (cystic fibrosis patients)

 Inhibitors of Protein Synthesis

• Tetracyclines
– Broad spectrum
• Gram neg and Gram pos
• Interferes with tRNA attachment to ribosome
• Do not interfere with mammalian ribosomes
– Produced by Streptomyces
• Penetrate body tissues
• Often treat rickettsias and chlamydias (intracellular
parasites), UTIs, syphilis, and gonorrhea
– Doxycycline
• Longer retention in body

 Inhibitors of Protein Synthesis

• Macrolides
– Gram-positives
• Binds 50S; prevents translocation
– Erythromycin
• What does erythro- mean?
– Clarithromycin (Biaxin®)
• Newer, broader range, penetrates tissues better
• Diarrhea is a common symptom
– Mega Colon Cleanse

 Injury to the Plasma Membrane

• Drugs block fatty acid synthesis
• Triclosan
– In some toothpaste and antibacterial soap
• Polymyxin B
– Topical
– Combined with bacitracin and neomycin in
over-the-counter preparation



Inhibitors of Nucleic Acid Synthesis
• Rifamycin
– Inhibits RNA synthesis
– Antituberculosis
• Fluoroquinolones
– Inhibits DNA gyrase
• Gyrase is needed to unwind DNA to make gene
products
– Ciprofloxacin (Cipro)
• Especially effective against anthrax infections
– Good for urinary tract infections

 Competitive Inhibitors

• Sulfonamides (sulfa drugs)

– Bacteria must synthesize PABA to make DNA
– Inhibit folic acid synthesis
• Folic acid is needed to make purine and pyrimidine bases of
nucleic acids
– Humans eat folic acid
• The drug doesn’t effect humans
– Broad spectrum
– Bacteriostatic
– Silver sulfadiazine
• Burn wounds

Competitive Inhibitors
 Antifungal Drugs
• Inhibition of ergosterol synthesis
– Primary sterol in fungal cell walls
– (animals use cholesterol, not ergosterol)
• Miconazole
– Inhibits ergosterol synthesis
• Tolnaftate
– Action unknown
• Tea Tree Oil
– Australian-type

 The Disk-Diffusion Method

• Kirby-Bauer test
– Confluent lawn of bacterial growth
– Drug-impregnated disc placed on surface of agar plate
• Drugs diffuse out over medium into agar
– The farther it diffuses, the lower the concentration
• A zone of inhibition is formed
– Diameter measured
– Generally, the bigger the zone, the more susceptible
– Recorded: sensitive, intermediate, resistant
• HOWEVER, some drugs have poor solubility in
water
– Give false result of resistance

The Disk-Diffusion Method

 Antibiotic Synergism

Synergism occurs when the effect of two drugs

together is greater than the effect of either alone
 The E Test
• Determines Minimal Inhibitory Concentration
– Lowest concentration that prevents visible
bacterial growth
• Plastic strips are coated with concentration
gradients
– Like disc-diffusion, but with many concentrations

The E Test
 Broth Dilution Test
• Make a series of decreasing concentrations of
broths
• Broths are then inoculated with bacteria
• If bacteria don’t grow, then concentration is
effective
• Minimal bactericidal concentration
– Wells not showing growth are cultured without presence
of drug
• If bacteria grow, then drug was bacteriostatic
• If no bacteria grow, then drug was bactericidal

Broth Dilution Test

 Antibiotic Resistance
• A variety of mutations can lead to antibiotic
resistance
• Mechanisms of antibiotic resistance
– Enzymatic destruction of drug
– Prevention of penetration of drug
– Alteration of drug's target site
– Rapid ejection of the drug
• Resistance genes are often on plasmids that can be
transferred between bacteria

Antibiotic Resistance
 Antibiotic Resistance

• Misuse of antibiotics selects for resistance mutants. Misuse

includes:
• Using outdated or weakened antibiotics
• Using antibiotics for the common cold (viral)
• Using antibiotics in animal feed
• Failing to complete the prescribed regimen
• Using someone else's leftover prescription
• Using hand sanitizer every minute like a tool
• Using antibacterial soap every minute like a tool
– Especially if you just soap up your hands and don’t SCRUB
– SCRUBBING IS WHAT WORKS TO CLEAN HANDS
– ANTIBACTERIAL SOAP IS UNNECESSARY UNLESS
• You are a healthcare worker or lab researcher

 Future of Chemotherapeutic Agents

• siRNA
– Complementary
RNA that binds
mRNA to inhibit
translation
– Promising for
viral treatments