‫بسم ال الرحمن الرحيم‬

Mechanisms of actions of antimicrobial drugs

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Introduction
• Strategies to combat microbes:
• • • • • Public health measures Screening procedures, early detection Vaccines Drugs Drugs therapy targets selective differences between microbes and host cells
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Selective targeting of microorganisms
• Targets unique to microbes & lack in host cells: • Bacterial cell wall contains peptidoglycan strands e.g. Penicillins ↓ cross linking of peptidoglycans • Fungal cell membrane contains ergosterol e.g. Azole antifungals ↓ ergoserol synthesis • Targets similar to microbes & have quantitative differences from the host cells: • Different enzyme or receptor isoform e.g.trimethoprim inhibits bacterial DHFR & pyrimethamine inhibits malarial DHFR • For selective targeting important to understand mechanisms of actions of drugs
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Mechanism of action of antimicrobials (overview)
Inhibitors of cell wall synthesis Inhibitors of cell membrane synthesis
Pteridine Folinic acid Folic acid Pyrimidines

Ribosome
50S & 30S

PABA

Purines

mRNA

DNA Inhibitors of protein synthesis Inhibitors of DNA & cell division
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Inhibitors of cell wall synthesis
• A. Inhibitors of synthesis of peptidoglycan: • Bacterial cell wall contains peptidoglycan strands • Peptidoglycan is composed of UDP-N-acetyl-muramic acid, UDP-N-acetyl-glucosamine & a pentapeptide • Cycloserine, resembels to alanine & inhibits addition of alanine into peptide chain • Vancomycin inhibits transglycosidase enzyme & prevents peptidoglycan chain elongation

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Inhibitors of cell wall synthesis (Cont.)
• B. Inhibitors of cross linking of peptidoglycan strands: • Beta-lactam antibiotics: e.g. Penicillins & cephalosporins • Inhibit transpeptidase enzyme, involved in cross linking of peptidoglycan strands • Also called as transpeptidation reaction & strengthens cell wall. • Defects in the synthesis of cell wall cause wholes in cell wall & chnages in permeability • Leading to bacterial swelling & lysis • Hence cell wall synthesis inhibitors are bactericidal
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Inhibitors of cell wall synthesis (Cont.)
• Inhibitors of mycobacterial cell wall: • Mycobacterial cell wall is composed of arabinogalactan chains • Ethambutol inhibits arabinosyl-transferase enzyme involved in addition of arabinose in arabino-galactan chains • Isoniazid & pyrazinamide inhibit synthesis of mycolic acid for mycobacterial cell wall by inhibiting fatty-acid synthase enzyme (FAS)

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Inhibitors of cell membranes
• Inhibitors of fungal cell membrane • A. Inhibitors of ergosterol synthesis • Azole anti-fungals: e.g. fluconazole inhibit fungal P-450 enzyme (14α- demethylase) • Defects in synthesis causes wholes in cell membrane • B. Ergosterol binding compounds • Polyene anti-fungals: e.g. amphotericin B Bind to ergosterol in the cell membrane & increase membrane permeability • Leakage of essential elements, cell lysis, fungicidal
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Inhibitors of protein synthesis
• Protein synthesis takes place in ribosomes • Bacterial ribosome consists of 30S & 50S sub-units whereas, mammalian ribosomes have 40S & 60S • Tetracyclines ↓ 30S ribosomal subunit & are bacteriostatic • Macrolides & chloramphenicol ↓ 50S ribosomal subunit & are bacteriostatic • Aminoglycosides ↓ 30S ribosomal subunit, but some abnormal proteins insert in cell membrane & form pores, thus become bactericidal • Rifampin ↓ DNA dependent RNA polymerase & is bactericidal

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Inhibitors of DNA & cell division
• Antimicrobial drugs can affect microbial DNA & cell division in following ways: • Inhibit DNA synthesis Inhibitors of folate synthesis Inhibitors of thymidylate synthesis • Inhibit DNA replication • Inhibit microtubules & mitosis
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Inhibitors of DNA synthesis
Folate precursors (PABA, pteridine, glutamic acid) - Sulfonamides & sulfones Trimethoprim Folic acid Folinic acid Purines Pyrimidines (adenine & guanine) (cytosine & thymine) Ribonucleotides Deoxy-ribonucleotides DNA

-

Flucytosine

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Inhibitors of DNA synthesis (Cont.)
• A. Inhibitors of folate metabolism • Folic acid (dihydro-folate) synthesis inhibitors: Sulfonamides & sulfones inhibit dihydro-pteroate synthase (DHPS) enzyme • Folinic acid (tetrahydro-folate) synthesis inhibitors: Trimethoprim inhibits dihydro-folate reductase (DHFR) enzyme (bacterial DHFR more than human)

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Inhibitors of DNA synthesis (Cont.)
• Synergism of folic acid (dihydrofolate) & folinic acid (tetrahydro-folate) synthesis inhibitors: • Sulfamethoxazole + PABA + pteridine Trimethoprim Glutamic acid Dihydro-pteroate (anti-bacterial) Sulfamethoxazole ↓ synthase
Folic acid Dihydro-folate Trimethoprim ↓ reductase Folinic acid DNA
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Inhibitors of DNA synthesis (Cont.)
• • • • B. Inhibitors of thymidylate synthesis Flucytosine: is a cytosine analogue Converted in fungi & GI flora to 5-fluorouracil (5-FU) 5-FU converted to 5-FdUMP
dUMP TS dTMP DNA

• 5-FdUMP incorporated to DNA & inhibits thymidylate synthase (TS) • ↓ DNA synthesis

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Inhibitors of DNA replication & mitosis
• During cell division chromosomes line up in the equator of cell • Each divides into two (DNA- replication) by topoisomerases (topoisomerase I to IV) • Each part is pulled to opposite pole of the cell by mitotic spindles made up of microtubules • Quinolones inhibit bacterial topisomerase-II [ in gram (-) bacteria] & topoisomerase IV [in gram (+) bacteria], inhibit DNA replication & are bactericidal • Griseofulvin inhibits microtubule assembly, mitotic spindle formation, cell division & is bacteriostatic
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