ANTIMICROBIAL DRUGS

Antimicrobial Drugs
I.

Terminology II. History III. Spectrum of antimicrobial activity IV. Action of antimicrobial drugs V. A survey of commonly used antimicrobial drugs VI. Test to determine chemotherapy VII. Effectiveness of Drugs

I. Terminology
Antimicrobial

drugs are chemotherapeutic drugs. Two categories: antibiotics Antimicrobial drugs produced by microorganisms. synthetic drugs Antimicrobial drugs synthesized in the lab. Antibacterial synthetic drugs Antifungal synthetic drugs Antiviral agents

II. History
Paul

Ehrlich (1910)

Knew: some dyes bind better to microorganisms than to animal cells. He bound a poison to a dye. Gave it to rabbits with syphilis. The pathogen (Treponema pallidum) was killed: the host was not harmed. Was a magic bullet

II. History, cont.

Alexander

Fleming (1928)

In England, noticed that S. aureus did not grow around a colony of mold on agar The mold was Penicillium notatum. He isolated the inhibitory substance. Called it penicillin. Penicillin was unstable.

II. History, cont.

Florey

and Chain (1940) In England Resumed study of penicillin Isolated and purified penicillin USA became involved Penicillin used during WWII

II. History, cont.

Penicillin

is an antibiotic. Antibiotic is from antibiosis, meaning against life. antibiotic A substance that is produced by one microorganism (a bacterium or fungus) that kills or inhibits the growth of another microorganism.

II. History, cont.

Major

producers of antibiotics discovered throughout the years: Molds Penicillium Cephalosporium Bacteria Streptomyces Bacillus

III. Spectrum of antimicrobial activity

Principle

of selective toxicity: a drug should selectively kill or prevent growth of a pathogen, but not of host cells. Differences to be considered:
1. Between procaryotic pathogen and eucaryotic host 2. Between eucaryotic pathogen and eucaryotic host 3. Between eucaryotic host and viruses

III. Spectrum of antimicrobial activity, cont.

Narrow

spectrum drugs affect only Grampositive cells or only Gram-negative cells. Broad spectrum drugs affect both Grampositive and Gram-negative cells. The normal flora is affected, too.

Fig. 13-1

IV. Action of antimicrobial drugs

Have

selective toxicity They act at a specific site, unlike disinfectants. Are bactericidal or bacteriostatic Or, are fungicidal or fungistatic

IV. Action of antimicrobial drugs, cont.

A.

Inhibition of cell wall synthesis B. Inhibition of protein synthesis C. Injury to plasma membrane D. Inhibition of nucleic acid synthesis E. Inhibition of synthesis of essential metabolites (F. Antifungal drugs) (G. Antiviral drugs)

IV. Action of antimicrobial drugs, cont.

A.

Inhibition of cell wall synthesis

Penicillins and cephalosporins stops synthesis of wall by preventing cross linking of peptidoglycan units. Bacitracin and vancomycin also interfere here. Excellent selective toxicity

IV. Action of antimicrobial drugs, cont.

B.

Inhibition of protein synthesis Due to differences in ribosomes Eucaryotic cells have 80S (60S + 40S subunits) ribosomes. Procaryotic cells have 70S (50S + 30S subunits) ribosomes. Examples: Chloramphenicol and erythromycin bind to the 50S subunit. Tetracyclines bind to the 30S subunit.

IV. Action of antimicrobial drugs, cont.

C.

Injury to plasma membrane Bind to membrane, alter permeability, and cause leakage Or, antifungal antibiotics bind to sterols in membrane and disrupt membrane. Animal cell membranes have cholesterol. Fungal membranes have ergosterol. Procaryotic cell membranes have no sterols. Example: amphotericin B

IV. Action of antimicrobial drugs, cont.

D.

Inhibition of nucleic acid synthesis Stop DNA replication Many antiviral drugs do this. Example: AZT Or stop RNA synthesis Example: rifampin

IV. Action of antimicrobial drugs, cont.

E.

Inhibition of synthesis of essential metabolites A drug mimics a normal metabolite and acts as a competitive inhibitor. Enzyme of cell recognizes the drug instead of the normal metabolite Pathway stops. Example: sulfa drugs are similar to PABA, para aminobenzoic acid.

IV. Action of antimicrobial drugs, cont.

A.

Inhibition of cell wall synthesis B. Inhibition of protein synthesis C. Injury to plasma membrane D. Inhibition of nucleic acid synthesis E. Inhibition of synthesis of essential metabolites

Fig. 13-2

V. A survey of commonly used antimicrobial drugs

A.

Inhibitors of cell wall synthesis Over 50 penicillins Are bactericidal Some are naturally produced. Some are produced semisynthetically. Each has a unique side chain. Inhibit transpeptidase activity, and peptide crosslinking in wall

1. The penicillins

Penicillins
O S R C NH O CH C CH N C CH3 CH COOH CH3

B-lactam ring

Common nucleus

V. A survey of commonly used antimicrobial drugs, cont.

1. The penicillins, cont. Side chain (R group) affects: spectrum of penicillin pH stability sensitivity to penicillinase, a betalactamase

O R C NH O CH C

Penicillins
S CH N C CH3 CH COOH CH3

penicillinase or low pH

O S R C NH O CH C CH N C

CH3 CH3 CH COOH

OH H

Penicilloic acid

Penicillins
O S CH3 C CH3 O C N CH COOH

Penicillin G

CH2

NH

CH

CH

Penicillin V

CH2

Common nucleus

Ampicillin

CH NH2

Common nucleus

V. A survey of commonly used antimicrobial drugs, cont.

1. The penicillins, cont. Natural penicillins are secreted by the mold Penicillium. Prototype is penicillin G Effective against Gram-positive cells, e.g., staph and strep Is pH sensitive. Therefore not given orally. Penicillin V is acid stable. Both are susceptible to penicillinase

V. A survey of commonly used antimicrobial drugs, cont.

1. The penicillins, cont. Semisynthetic penicillins Produce by growing Penicillium in culture so that only the nucleus is synthesized. Attach R group in lab. Or, grow Penicillium, extract natural penicillin, remove R group, and attach wanted R group. Have broader spectrum. Are effective against Gram-negative cells, too. Are not resistant to penicillinases

V. A survey of commonly used antimicrobial drugs, cont.

A.

Inhibitors of cell wall synthesis 1. Penicillins 2. Monobactams 3. Cephalosporins More than 70 available Similar in structure to the penicillins Are resistant to penicillinases Have broader spectrum than penicillins Not pH sensitive

Penicillins and Cephalosporins

O S R C NH O CH C CH N C CH3 CH COOH CH3

B-lactam ring O

Penicillin nucleus
S

NH O

CH C

C N C

C H2 C CH2 O

O C CH3

Cephalosporin nucleus

V. A survey of commonly used antimicrobial drugs, cont.

A. Inhibitors of cell wall synthesis, cont. 4. Carbapenems, a new group Very broad spectrum Have a beta-lactam ring 5. Bacitracin Is used topically Effective mainly against Gram + cells 6. Vancomycin Used to treat penicillinase-producing staphylococc. Very narrow spectrum

V. A survey of commonly used antimicrobial drugs, cont.

A. Inhibitors of cell wall synthesis, cont. 7. Isoniazid Is not an antibiotic, but a synthetic drug Effective against Mycobacterium tuberculosis. Stops synthesis of mycolic acids. Bacteriostatic 8. Ethambutol Effective against M. tuberculosis Stops synthesis of mycolic acid Bacteriostatic

V. A survey of commonly used antimicrobial drugs, cont.

A.

Inhibitors of cell wall synthesis B. Inhibitors of protein synthesis

V. A survey of commonly used antimicrobial drugs, cont.

B.

Inhibitors of protein synthesis. 1. Aminoglycosides Bactericidal Stop protein synthesis Example: Streptomycin Used for tuberculosis Side effect: damages 8th cranial nerve Other examples: neomycin & gentamicin

V. A survey of commonly used antimicrobial drugs, cont.

B.

Inhibitors of protein synthesis, cont. 2. Tetracyclines Bacteriostatic Very broad spectrum Produced by Streptomyces Used in urinary tract infections Causes tooth discoloration, liver & kidney damage

Tetracycline
H3C H3C OH N CH3 OH OH OH O OH O C ONH2

V. A survey of commonly used antimicrobial drugs, cont.

B. Inhibitors of protein synthesis, cont. 3. Chloramphenicol Bacteriostatic Broad spectrum Stops protein synthesis by binding to 50S subunit. Stops elongation. Side effect is aplastic anemia. Used for some meningitis cases and typhoid fever

V. A survey of commonly used antimicrobial drugs, cont.

B. Inhibitors of protein synthesis, cont. 4. Macrolides Bacteriostatic Have macrocyclic lactone ring Example: erythromycin Inhibits protein synthesis by binding to the 50S subunit Narrow spectrum. Affects mainly Grampositive cells but some Gram- negative cells, too.

V. A survey of commonly used antimicrobial drugs, cont.

A.

Inhibition of cell wall synthesis B. Inhibition of protein synthesis C. Injury to plasma membrane D. Inhibition of nucleic acid synthesis E. Inhibition of synthesis of essential metabolites (F. Antifungal drugs) (G. Antiviral drugs)

V. A survey of commonly used antimicrobial drugs, cont.

C.

Injury to plasma membrane Polymyxin B Bactericidal Effective against Gram-negative bacteria.

V. A survey of commonly used antimicrobial drugs, cont.

D.

Inhibition of nucleic acid synthesis 1. Rifamycins Example: Rifampin Bactericidal Stops transcription Used to treat tuberculosis and leprosy 2. Quinolones and fluoroquinolones Bactericidal

V. A survey of commonly used antimicrobial drugs, cont.

E.

Inhibition of synthesis of essential metabolites Sulfonamides. Also called the sulfa drugs. Are bacteriostatic. Discovered by Domagk in the 1930s He gave animals with a streptococcus infection the dye prontosil. The animal was cured. Put prontosil in test tube with streptococcus, cells were not killed

V. A survey of commonly used antimicrobial drugs, cont.

Sulfonamides, cont. Prontosil breaks down to form sulfanilamide. Sulfanilamide is similar in shape to PABA. PABA is part of folic acid. Sulfanilamide is a competitive inhibitor of enzyme that incorporates PABA into folic acid. Result: folic acid synthesis stops

NH2 H2N N N H2N NH2 NH2

NH2

+
SO2NH2

SO2NH2

Prontosil

Sulfanilamide

NH2

NH2

SO2NH2

COOH

Sulfanilamide

Para-aminobenzoic acid (PABA)

Folic Acid
OH N H2N N N N C H2 H N O C H N COOH CH C H2 C H2

PABA

C OOH

V. A survey of commonly used antimicrobial drugs, cont.

Sulfonamides, cont. Sulfanilamide is an antimetabolite. Is a synthetic drug Humans are not affected because we get folic acid from our diets. Excellent selective toxicity Many sulfa drugs are available.

III. Action of Antimicrobial Drugs

A.

Inhibition of cell wall synthesis B. Inhibition of protein synthesis C. Injury to plasma membrane D. Inhibition of nucleic acid synthesis E. Inhibition of synthesis of essential metabolites (F. Antifungal drugs) (G. Antiviral drugs)

V. A survey of commonly used antimicrobial drugs, cont.

F.

Antifungal drugs 1. Polyenes Amphotericin B and nystatin Secreted by Streptomyces Are fungicide Combines with sterols in membranes Used topically due to its toxicity

V. A survey of commonly used antimicrobial drugs, cont.

F.

Antifungal drugs, cont. 2. Imidazoles and triazoles Interfere with sterol synthesis in fungi Used topically or orally 3. Griseofulvin Produced by Penicillium Taken orally. Accumulates in keratin, and is found in nails, hair, skin. Interferes with mitosis

V. A survey of commonly used antimicrobial drugs, cont.

G.

Antiviral drugs Antibiotics do not act on viruses. Difficult to get good selective toxicity against viruses Relatively few approved for use in USA Three major groups 1. Amantadine 2. Thiosemicarbazones 3. Base analogs

V. A survey of commonly used antimicrobial drugs, cont.

G.

Antiviral drugs, cont. 1. Amantadine Acts against the influenza virus Prevents the virus from entering the cell or from uncoating from capsid once inside the cell Given to elderly

V. A survey of commonly used antimicrobial drugs, cont.

G.

Antiviral drugs, cont. 2. Thiosemicarbazones Example: Methisazone Used for small pox Stops translation of viral mRNA 3. Base analogs or nucleoside analogs a. Idoxuridine is an analog of thymidine. Used in eye for HSV1 infection

O N O HOCH 2 C N C C CH3 O HOCH 2 O N C

O I C

OH

OH

Thymidine

Idoxuridine

V. A survey of commonly used antimicrobial drugs, cont.

3. Base analogs, cont. b. Acyclovir, an analog of guanine Used to treat Herpes virus infections Only viral enzyme acts on acyclovir Stops viral DNA synthesis Is only effective in cells infected with a Herpes virus

V. A survey of commonly used antimicrobial drugs, cont.

3. Base analogs, cont. c. AZT, azidothymidine An analog of thymidine Used to treat AIDS patients Stops RNA dependent DNA polymerase of HIV

O N ' '
5'

O C CH3 O HOCH 2 N C N C CH3

O HOCH 2
4' 3'

O
1' 2'

5'

O
1' 2'

4' 3'

OH

N3

Thymidine Azidothymidine (AZT)

V. A survey of commonly used antimicrobial drugs, cont.

3. Base analogs, cont. Also used to treat AIDS patients ddI, dideoxyinosine ddC, dideoxycytosine Others 4. Enzyme inhibitors Protease inhibitors to stop HIV replication Inhibitors of reverse transcriptase

Antimicrobial Drugs
I.

Terminology II. History III. Spectrum of antimicrobial activity IV. Action of antimicrobial drugs V. A survey of commonly used antimicrobial drugs VI. Test to determine chemotherapy VII. Effectiveness of Drugs

VI. Test to determine chemotherapy

Disk

diffusion method: The Kirby Bauer Technique

Isolate pathogen in pure culture Spread onto agar Put discs containing antibiotic on agar Incubate plates Look for zones of inhibition

Fig. 13-9

VII. Effectiveness of drugs

Bacteria

may become resistant to drugs. Occurs via: 1) Mutation 2) Acquiring a plasmid, an R factor, that contains genes coding for enzymes that make the cell resistant to a specific antibiotic

VII. Effectiveness of drugs, cont.

Bacterial

resistance to drugs occurs by: 1. Inactivation of drug Cell secretes an enzyme that hydrolyzes antibiotic 2. Preventing drug entry into cell 3. Alteration of drug target inside cell

VII. Effectiveness of drugs, cont.

Significance

of drug resistance: May be some cells in a population that are resistant. In absence of drugs, these cells are low in number. In presence of drugs, sensitive cells die, resistant cells replicate. Get huge populations of resistant cells.

Fig. 13-8

Summary
Antimicrobial

drugs include the antibiotics and the synthetic drugs. Antimicrobial drugs exhibit some selective toxicity. Therefore, these drugs act more specifically than do disinfectants and antiseptics.