The Microbial World

Lectures in Microbiology

Antimicrobial Agents Used in the Treatment of Infectious Disease
Introduction Most microbiologists distinguish two groups of antimicrobial agents used in the treatment of infectious disease: antibiotics, which are natural substances produced by certain groups of microorganisms, and chemotherapeutic agents, which are chemically synthesized. A hybrid substance is a semisynthetic antibiotic, wherein a molecular version produced by the microbe is subsequently modified by the chemist to achieve desired properties. Furthermore, some antimicrobial compounds, originally discovered as products of microorganisms, can be synthesized entirely by chemical means. In the medical and parmaceutical worlds, all these antimicrobial agents used in the treatment of disease are referred to as antibiotics, interpreting the word literally. The modern era of antimicrobial chemotherapy began in 1929, with Fleming's discovery of the powerful bactericidal substance, penicillin, and Domagk's discovery in 1935 of synthetic chemicals (sulfonamides) with broad antimicrobial activity. In the early 1940's, spurred partially by the need for antibacterial agents in WW II, penicillin was isolated and purified and injected into experimental animals, where it was found not only to cure infections but also to possess incredibly low toxicity for the animals. This fact ushered into being the age of antibiotic chemotherapy, and an intense search for similar antimicrobial agents of low toxicity to animals that might prove useful in the treatment of infectious disease. The rapid isolation of streptomycin, chloramphenicol and tetracycline soon followed, and by the 1950's, these and several other antibiotics were in clinical usage.

Microorganisms that Produce Antibiotics

The bacterial colonies at 10 o'clock, 2 o'clock and 8 o'clock on this agar plate are producing antibiotics that inhibit encroachment by the mold which is growing out from the center.

Most of the natural antibiotics that are being used in agriculture and medicine are produced by three unrelated groups of microbes, including eucaryotic molds and two types of spore-forming bacteria. However, many culturable, and some non culturable microbes, have been shown to produce various substances that inhibit other organisms that grow in their space. If we consider antibiotics as secondary metabolites of microbes, it narrows the field to the handful of microbes discussed below. 1. Penicillium and Cephalosporium molds produce beta-lactam antibiotics such as penicillin and cephalosporin and their relatives. They also produce the base molecule for development of semisynthetic beta-lactam antibiotics, such as amoxacillin and ampicillin. Beta-lactams are used to treat about one-third of outpatients with bacterial infections. The natural habitat of molds is soil. And although sex is sometimes involved, they reproduce by spore formation. They are foremost in their abilities to degrade organic matter, and they play their most important role in natures in biodegradation and the carbon cycle. Most of us know that molds will grow on nearly anything that is organic and moist, so they are also responsible for a lot food spoilage as well as decomposition of our structural materials and textiles. "Nothing is forever", with molds around.

They come from a phylum of Bacteria. Some of the representative family include such diverse bacteria as Actinomyces. Streptomyces. mainly Streptomyces species. Propionibacter. Corynebacterium. macrolides (erythromycin and its relatives). Actinomycetes are a group of branched bacteria that reproduce by spore formation.Three colonies of a Penicillium mold growing on an agar medium. and they are landed in Order Actinomycetales. Nocardia. by these bacteria . aminoglycosides (streptomycin and its relatives). chloramphenicol. Actinomycetes are the mainstay of the antibiotics industry. rifamycins. called geosmins. Most actinomycetes are inhabitants of the soil. produce tetracyclines. Micromonospora and Frankia. Actinobacteria. ivermectin. The green fuzzy appearance is the asexual spores of the fungus. 2. Actinomycetes. and most other clinically-useful antibiotics that are not beta-lactams. The characteristic odor of damp soil is due to the production of substances.

What could be going on? Courtesy of Jerry Ensign Department of Bacteriology. cereus produces zwittermicin." 3. aerobic bacteria that live in the soil. produce polypeptide antibiotics (e. Bacillus species have the relatively rare ability to form a type of resting cell called an endospore.g.Two different actinomycetes were spotted in the center of the agar plate about two centimeters apart. subtilis. "Chance favors the prepared mind. They play an important ecological role in aerobic decomposition. and B. Bacillus species. . This peculiar pattern of growth was observed after a 10-day incubation period. polymyxin and bacitracin). polymyxa and B. biodegradation and mineral recycling. rod-shaped. Bacilli are Grampositive. such as B.

if not essential. The maintenance of a substantial component of the bacterial genome devoted solely to the synthesis of an antibiotic leads one to conclude that the antibiotic is affords them some nutritional or spatial advantage in their habitat by antagonizing the competition. Most of the microorganisms that produce antibiotics are resistant to the action of their own antibiotic. although the organisms are affected by other antibiotics. but the answer may be in the obvious . how or why bacteria are resistant to their . at least in the Northern Hemisphere where I have seen it. Antibiotics tend to be rather large. Antibiotics are secondary metabolites and they are produced at the same time that the cells begin their sporulation processes. to the survival of these organisms in their natural habitat. and their antibiotic may be effective against closely-related strains.A swirl of Bacillus mycoides colonies growth amidst other bacteria and molds from the soil. It is not known why these microorganisms produce acts as some sort of hormone or signal molecule associated with sporulation or dormancy or germination. These organisms all have in common that they live in soil and they form some sort of a spore or resting structure. In most cases. or it may be in the subtle . complicated organic molecules and may require as many as 30 separate enzymatic steps to synthesize. The swirls are always counterclockwise.

including streptomycin and the tetracyclines. A clinically-useful antibiotic should have as many of these characteristics as possible. In practice.. they are referred to as limited spectrum. Antibiotics such as the flouroqinolones (e. target bacterial protein synthesis because bacterial ribosomes (termed 70S ribosomes) are different from the ribosomes (80S) of humans and other eucaryotic organisms. or bacterial nucleic acid synthesis. ciprofloxacin) inhibit procaryotic (not eucaryotic) DNA replication. The selective toxicity of antibiotics is brought about by finding vulnerable targets for the drug in the microbe that do not exist in the animal (eucaryote) that is given the drug. and rifamycins inhibit bacterial (not eucaryotic) DNA transcription. Other antibiotics. the beta lactam antibiotics (penicillin and its relatives) inhibit peptidoglycan synthesis in the cell wall. Antibiotics must have Selective Toxicity for the Microbe Several hundreds of compounds with antibiotic activity have been isolated from microorganisms over the years. If effective against a single organism or disease. -It should be nontoxic to the host and without undesirable side effects. bacterial protein synthesis. but it may be worth pondering or studying if we are to understand the cellular and molecular basis of drug resistance in pathogens.the ratio of the toxic dose (to the patient) to the therapeutic dose (to eliminate the infection). but only a few of them are clinically-useful. the safer is the drug (antibiotic) for human use. Characteristics of Antibiotics Antibiotics may have a cidal (killing) effect or a static (inhibitory) effect on a range of microbes. this is expressed by a drug's therapeutic index (TI) .own antibiotics is also unknown. -It should have a wide spectrum of activity with the ability to destroy or inhibit many different species of pathogenic organisms. The selective toxicity of antibiotics means that they must be highly effective against the microbe but have minimal or no toxicity to humans. For example. The larger the index. the most important property of an antimicrobial agent is its selective toxicity. that the agent acts in some way that inhibits or kills bacterial pathogens but has little or no toxic effect on the patient. Humans have neither a cell wall nor peptidoglycan and so are unaffected by the action of the drug. The reason for this is that only compounds with selective toxicity can be used clinically. . Antibiotics effective against procaryotes that kill or inhibit a wide range of Gram-positive and Gram-negative bacteria are said to be broad spectrum. The range of bacteria or other microorganisms that is affected by a certain antibiotic is expressed as its spectrum of action. i. which are unique in some ways to bacteria. From a patient point of view.g. If effective mainly against Gram-positive or Gram-negative bacteria. they are narrow spectrum.e. Most antibiotics in clinical usage are directed against bacterial cell wall synthesis.

Cephalothin Semisynthetic beta-lactams Ampicillin. -It should be chemically-stable (have a long shelf-life). -It should not eliminate the normal flora of the host. Amoxicillin Augmentin is Streptomyces clavulanic acid clavuligerus plus Amoxicillin Gram-positive and Gramnegative bacteria Gram-positive and Gramnegative bacteria Clavulanic Acid Monobactams Aztreonam Gram-positive Chromobacterium and Gramviolaceum negative bacteria Streptomyces cattleya Gram-positive and Gramnegative bacteria Carboxypenems Imipenem . -It should be inexpensive and easy to produce. -It should be able to reach the part of the human body where the infection is occurring. spectrum and mode of action.-It should be nonallergenic to the host. Kinds of Antimicrobial Agents and their Primary Modes of Action The table below is a summary of thetypes or classes of antibiotics and their properties including their biological source. -Microbial resistance is uncommon and unlikely to develop. Classes of Antibiotics and their Properties Chemical class Examples Biological source Penicillium notatum and Cephalosporium species Spectrum (effective against) Gram-positive bacteria Mode of action Inhibits steps in cell wall (peptidoglycan) synthesis and murein assembly Inhibits steps in cell wall (peptidoglycan) synthesis and murein assembly Inhibitor of bacterial betalactamases Inhibits steps in cell wall (peptidoglycan) synthesis and murein assembly Inhibits steps in cell wall (peptidoglycan) synthesis and Beta-lactams (penicillins and cephalosporins) Penicillin G.

synthesis) Legionella. (protein erythreus Neisseria. Gramnegative bacteria Inhibit translation Streptomyces not enterics. anaerobic synthesis) Bacteroides Gram-positive bacteria. synthesis) Pseudomonas Amycolatopsis Inhibits steps in Gram-positive orientalis (formerly murein bacteria. Mycoplasma Damages Gram-negative Bacillus polymyxa cytoplasmic bacteria membranes Inhibits steps in murein Gram-positive Bacillus subtilis (peptidoglycan) bacteria biosynthesis and assembly Inactivate Streptomyces Fungi membranes nodosus (Histoplasma) containing sterols Inactivate Streptomyces Fungi (Candida) membranes noursei containing sterols Streptomyces Gram-positive Inhibits mediterranei and Gramtranscription .Aminoglycosides Streptomycin Gentamicin Glycopeptides Vancomycin Lincomycins Clindamycin Macrolides Erythromycin. esp. Azithromycin Polypeptides Polymyxin Bacitracin Polyenes Amphotericin Nystatin Rifamycins Rifampicin murein assembly Inhibits Gram-positive Streptomyces translation and Gramgriseus (protein negative bacteria synthesis) Gram-positive Inhibits and GramMicromonospora translation negative bacteria species (protein esp. designated (peptidoglycan) Staphylococcus Nocardia biosynthesis and aureus orientalis) assembly Gram-positive Inhibits and GramStreptomyces translation negative bacteria lincolnensis (protein esp.

in the patient. Rickettsias Gram-positive and Gramnegative bacteria. Gantrisin.Tetracyclines Tetracycline Streptomyces species Semisynthetic tetracycline Doxycycline negative bacteria. . Recall that the target of an antibiotic should be unique to the bacterium and not found. analog of pyridoxine (Vit B6) Mycobacterium Anti-folate tuberculosis Antimicrobial Agents Used in the Treatment of Infectious Disease Examination of the foregoing table reveals that there are a handful of fundamental ways that antibacterial antibiotics work as therapeutic agents. or not accessible to the antibiotic.replication positive bacteria (Bacillus anthracis) Gram-positive Inhibits folic acid and Grammetabolism (antinegative bacteria folate) Mycobacterium Inhibits mycolic tuberculosis acid synthesis. Borrelia (bacterial RNA polymerase) Inhibit translation (protein synthesis) Inhibit translation (protein synthesis) Chloramphenicol Chloramphenicol Quinolones Nalidixic acid Streptomyces venezuelae synthetic synthetic Fluoroquinolones Ciprofloxacin Growth factor analogs Sulfanilamide. These are the most important targets in bacteria that have been exploited so far. Trimethoprim Isoniazid (INH) synthetic synthetic paraaminosalicylic acid (PAS) synthetic Inhibits Gram-positive translation and Gram(protein negative bacteria synthesis) Mainly GramInhibits DNA negative bacteria replication Gram-negative Inhibits DNA and some Gram. Mycobacterium tuberculosis Gram-positive and Gramnegative bacteria. Rickettsias Ehrlichia.

1. Interfere with protein synthesis. Chemically. which exploits differences between RNA polymerases and DNA replication strategies in bacteria and eucaryotes. 5. Penicillium and Cephalosporium. Attack is almost always ate the level of translation using 70S ribosomes in the translation machinery. This is usually brought about through the use of competitive chemical analogs for bacterial enzymatic reactions. Beta lactam antibiotics. Chemical structures of some beta-lactam antibiotics. Bacteria have murein in their cell walls. The beta lactam antibiotics are stereochemically related to D-alanyl-D-alanine. 2. However. Membrane inhibition or disruption doesn't work too well because of the similarities between eucaryotic and bacterial membranes. and murein (peptidoglycan) is essential to the viability of the bacterium. 3. the final cross-linking between between . which is a substrate for the last step in peptidoglycan synthesis. They exert their selective toxicity against bacteria because humans cells lack cell walls. and are correspondingly represented by the penicillins and cephalosporins. 4. Cell wall synthesis inhibitors Cell wall synthesis inhibitors generally inhibit some step in the synthesis of bacterial peptidoglycan. Interference with nucleic acid synthesis (RNA and DNA). carbapenem. 70S cytoplasmic ribosomes are absent in eucaryotes. Attack bacterial cell wall synthesis. these antibiotics contain a 4-membered beta lactam ring. monobactam. Inhibition of an essential metabolic pathway that exists in the bacterium but does not exist in the host. not found in the host. They are the products of two genera of fungi. the outer membrane of Gram-negative bacteria is a reasonable point of attack and some membrane inhibitors are included in the discussion below. Note the characteristic structure of the beta lactam ring. cephalosporin. Clockwise: penicillin.

Beta lactam antibiotics are bactericidal and require that cells be actively growing in order to exert their toxicity. meningitidis. Penicillins bind to and inhibit the carboxypeptidase and transpeptidase enzymes that are required for this step in peptidoglycan biosynthesis.. rate of clearance from blood. Penicillin G (Benzylpenicillin) is typically given by parenteral administration because it is unstable in the acid of the stomach. amoxicillin and ampicillin have broadened spectra against Gram-negative bacteria and are effective orally. whether they can be taken orally. and susceptibility to bacterial betalactamases. including Streptococcus pyogenes. . It is used against a wide range of Gram-positive bacteria.peptide side chains. A mold produces the main part of the molecule (6-aminopenicillanic acid). The semisynthetic beta-lactam. ability to cross the blood-brain barrier. However. Amoxicillin is usually the drug of choice within the class because it is better absorbed following oral administration than other beta-lactam antibiotics. are produced by fermentation of Penicillium chrysogenum. syphilis. such as penicillin G or penicillin V (benzyl penicillin). Susceptible Gram-negative organisms include non beta-lactamase producing strains of Haemophilus influenzae. They are effective against streptococci. Different beta lactams differ in their spectrum of activity and their effect on Gram-negative rods. amoxicillin. gonococci and staphylococci. as well as their toxicity. resistance to penicillinase. penicillin-sensitive Streptococcus pneumoniae. They are considered narrow spectrum since they are not effective against Gram-negative rods. such as increased spectrum of activity (effectiveness against Gram-negative rods). Semisynthetic penicillins first appeared in 1959. and pneumonia. methicillin is penicillinase-resistant. Natural penicillins. stability in the human body. this achieves higher tissue concentrations than orally-administered penicillins and this increases its antibacterial potential. except where resistance has developed. "PenG" may be used in treatment of bacterial endocarditis. Neisseria gonorrhoeae and N. non beta-lactamase producing strains of Staphylococcus aureus and Enterococcus faecalis. gonorrhea. which can be modified chemically by the addition of side chains. meningitis. etc. effectiveness when administered orally. Many of these compounds have been developed to have distinct benefits or advantages over penicillin G. It is susceptible to degradation by bacterial beta-lactamase enzymes so it may be given with calvulanic acid (below) to decrease its susceptibility.

The clavulanate is not an antimicrobial agent. It is a beta-lactamase inhibitor sometimes combined with semisynthetic beta lactam antibiotics to overcome resistance in bacteria that produce beta-lactamase enzymes. They are produced by species of Cephalosporium molds. there are 300 . Clavulanic acid is not an antibiotic. In the U. amoxicillin plus clavulanate is clavamox or augmentin. In allergic individuals the beta lactam molecule attaches to a serum protein and initiates an IgEmediated inflammatory response. clavamox.Clavulanic acid is a chemical sometimes added to a semisynthetic penicillin preparation. Cephalosporins are beta lactam antibiotics with a similar mode of action to penicillins. They are subject to degradation by some bacterial beta-lactamases. They are often used as penicillin substitutes against Gramnegative bacteria and in surgical prophylaxis.500 deaths annually due to penicillin allergy. which are structurally different so as not to induce allergy. Thus. It inhibits beta lactamase enzymes and has given extended life to penicillinase-sensitive beta lactams. A person who becomes allergic to penicillin usually becomes allergic to the cephalosporins and the carbapenems as well. Such individuals can still be treated with the monobactams. The core structure of cephalosporin. Two other classes of beta lactams are the carbapenems and monobactams.S. The latter are particularly useful for the treatment of allergic individuals. penicillins occasionally cause death when administered to persons who are allergic to them. The have a low toxicity and a somewhat broader spectrum than natural penicillins. aureus. Although nontoxic. but they tend to be resistant to beta-lactamases from S. . The structure of calvulanic acid. Most commonly it is combined with amoxicillin (above) as Augmentin (trade name) or the veterinary preparation. which otherwise inactivate the antibiotic. Substituent groups added at position X on the six-membered ring generates variants of the antibiotic.

is also inhibited. which requires the same carrier. as well as for the prevention of wound infections. Teichoic acid synthesis. and it competitively inhibits the racemase reaction that converts L-alanine to D-alanine and the synthetase reaction that joins two D-alanine molecules. Cycloserine enters bacterial cells by means of an active transport system for glycine and can reach a relatively high intracellular concentration. It is present in many topical antibiotic preparations. neomycin and polymyxin B. Tracy. However. It prevents cell wall growth by inhibiting the release of the muropeptide subunits of peptidoglycan from the lipid carrier molecule that carries the subunit to the outside of the membrane. along with its high affinity for susceptible enzymes. Cycloserine inhibits the early stages of murein synthesis where D-alanyl-D-alanine is added to the growing peptide side chain. The antibiotic resembles D-alanine in spatial structure. Haemophilus and Klebsiella. primarily against Gram-positive bacteria. it is given to "sterilize" the bowel prior to surgery. coli. including Pseudomonas aeruginosa. Bacitracin is a polypeptide antibiotic produced by Bacillus species.Aztreonam is a synthetic monocyclic beta lactam antibiotic (a monobactam) originally isolated from the bacterium Chromobacterium violaceum. . It is effective used topically. it is fairly toxic and has limited use as a secondary drug for tuberculosis. contains bacitracin. and since it is not absorbed by the gut. Bacitracin is a polypeptide antibiotic produced by the licheniformis group of Bacillus subtilis var. Bacitracin has a high toxicity which precludes its systemic use. The affinity of cycloserine for these enzymes is about a hundred times greater than that of D-alanine. It is not useful against Gram-positive bacteria but it has strong activity against a wide range of susceptible Gram-negative bacteria. E. It is used in ointment or cream form for topical treatment of a variety of localized skin and eye infections. A popular brand name Neosporin. This concentrating effect. enables cycloserine to function as a very effective antimicrobial agent.

and compounds that disorganize the membranes rapidly kill the cells. although the emergence of vancomycin-resistant organisms means that it is increasingly being displaced from this role by linezolid and the carbapenems. The only antibacterial antibiotics of clinical importance that act by this mechanism are the polymyxins. Vancomycin is a glycopeptide antibiotic used in the prophylaxis and treatment of infections caused by Grampositive bacteria. due to the similarities in phospholipids in eubacterial and eucaryotic membranes. For the treatment against tuberculosis. Polymyxin is effective mainly against Gram-negative bacteria and is usually limited to topical usage. However. Glycopeptides. inhibit both transglycosylation and transpeptidation reactions during peptidoglycan assembly. used only after treatment with other antibiotics had failed. Polymyxins bind to membrane phospholipids and thereby interfere with membrane . it has become important in clinical usage for treatment of infections by strains of Staphylococcus aureus that are resistant to virtually all other antibiotics (MRSA). Cell membrane inhibitors These antibiotics disorganize the structure or inhibit the function of bacterial membranes. produced by Bacillus polymyxa. such as the antibiotic vancomycin. However.Cycloserine is an oral broad spectrum antibiotic effective against tuberculosis. It has traditionally been reserved as a drug of "last resort". it is classified as a second line drug. The integrity of the cytoplasmic and outer membranes is vital to bacteria. Vancomycin is not effective against Gram-negative bacteria because it cannot penetrate their outer membrane. They bind to the muropeptide subunit as it is transferred out of the cell cytoplasm and inhibit subsequent polymerization reactions. this action is rarely specific enough to permit these compounds to be used systemically. by inhibiting cell wall synthesis of TB bacilli at the early stages of peptidoglycan synthesis.

streptomycin has been shown to prevent the initiation of protein synthesis by blocking the binding of initiator Nformylmethionine tRNA to the ribosome. They disrupt the structure of the bacterial cell membrane by interacting with its phospholipids. These antibiotics exert their activity by binding to bacterial ribosomes and preventing the initiation of protein synthesis. which distinguishes streptomycin and other aminoglycosides from most other protein synthesis inhibitors. Polymyxins are cationic detergent antibiotics. tobramycin and gentamicin. chloramphenicol. The aminoglycosides are products of Streptomyces species and are represented by streptomycin. the macrolides (e. carbenicillin and tobramycin resistant. Polymyxin B. and very poorly absorbed from the gastrointestinal tract. The most important antibiotics with this mode of action are the tetracyclines. with a general structure of a cyclic peptide with a long hydrophobic tail. Polymyxin antibiotics are highly neurotoxic and nephrotoxic. especially on Pseudomonas and coliform bacteria. streptomycin). Streptomycin binds to 30S subunit of the bacterial ribosome. Protein synthesis inhibitors Many therapeutically useful antibiotics owe their action to inhibition of some step in the complex process of protein synthesis. This evidently accounts for its antibacterial activity but does not explain its bactericidal effects. and the drug should only be given under close supervision in the hospital. Polymyxin is occasionally given for urinary tract infections caused by Pseudomonas strains that are gentamicin. Polymyxins also have antifungal activity. leaving them mainly in their 70S form and preventing the formation of polysomes.g. erythromycin) and the aminoglycosides (e.function. The overall effect of streptomycin seems to be one of distorting the ribosome so that it no longer can carry out its normal functions. Most have an affinity or specificity for 70S (as opposed to 80S) ribosomes. and they achieve their selective toxicity in this manner. specifically to the S12 protein which is involved in the initiation of protein synthesis. Polymyxins have a bactericidal effect on Gram-negative bacilli. Their attack is always at one of the events occurring on the ribosome and never at the stage of amino acid activation or attachment to a particular tRNA. Experimentally.g. . kanamycin. The balance between effectiveness and damage to the kidney and other organs is dangerously close. It also prevents the normal dissociation of ribosomes into their subunits.

Gentamicin and Tobramycin are mainstays for treatment of Pseudomonas infections. This prevents initiation of protein synthesis. Streptomycin stops bacterial growth by inhibiting protein synthesis. Specifically. Gentamicin is active against many strains of Gram-positive and Gram-negative bacteria. including actinomycin. interfering with the binding of formyl-methionyl-tRNA to the 30S subunit. Streptomyces griseus. Streptomycin is derived from the bacterium. . Waksman and his laboratory discovered several antibiotics. streptomycin. It was discovered in 1943. which is apparently lethal to the cells. Streptomycin has been used extensively as a primary drug in the treatment of tuberculosis. in the laboratory of Selman Waksman at Rutgers University. and neomycin. it binds to the 16S rRNA of the bacterial ribosome. including penicillin-resistant staphylococci.Streptomycin is the first aminoglycoside antibiotic to be discovered. Aminoglycosides have been used against a wide variety of bacterial infections caused by Grampositive and Gram-negative bacteria. An unfortunate side effect of aminoglycosides has tended to restrict their usage: prolonged use is known to impair kidney function and cause damage to the auditory nerves leading to deafness. They may have a bactericidal effect because this leads to cytoplasmic accumulation of dissociated 30S subunits. and was the first antibiotic to be used in treatment of tuberculosis. Kanamycin is active at low concentrations against many Gram-positive bacteria. Kanamycin and tobramycin have been reported to bind to the ribosomal 30S subunit and to prevent it from joining to the 50S subunit during protein synthesis. including some strains of Pseudomonas aeruginosa.

It is synthesized by Micromonospora. Tetracycline sparked the development of many chemically altered antibiotics and in doing so has proved to be one of the most important discoveries made in the field of antibiotics. Like all aminoglycosides. It is useful in treatment of infections caused by Pseudomonas aeruginosa. not by inhibiting protein synthesis. and in both cases. a genus of Gram-positive bacteria widely distributed in water and soil. Nonetheless. it is not systemically active because it is not absorbed to any appreciable extent from the small intestine. Neisseria meningitidis or Legionella pneumophila infections. The tetracyclines have a remarkably low toxicity and minimal side effects when taken by animals. used mostly to treat Gram-negative infections. chlortetracycline and doxycycline are the best known. The combination of their broad spectrum and low toxicity has led to their overuse and misuse by the medical community and the wide-spread development of resistance has reduced their effectiveness. This greatly enhances its antibacterial effectiveness and accounts for its specificity of action. It is a classic "broad-spectrum antibiotic" used to treat infections caused by Gram-positive and Gram-negative bacteria and some protozoa. most bacteria possess an active transport system for tetracycline that will allow intracellular accumulation of the antibiotic at concentrations 50 times as great as that in the medium. The tetracyclines consist of eight related antibiotics which are all natural products of Streptomyces. . Tetracyclines inhibit protein synthesis on isolated 70S or 80S (eucaryotic) ribosomes. when gentamicin is given orally. tetracyclines still have some important uses. since an effective concentration cannot be accumulated in animal cells.Gentamicin is an aminoglycoside antibiotic. Some newly discovered members of the tetracycline family (e. although some can now be produced semisynthetically or synthetically.g. such as the use of doxycycline in the treatment of Lyme disease. their effect is on the small ribosomal subunit. it is not used for Neisseria gonorrhoeae. The tetracyclines are a large family of antibiotics that were discovered as natural products of Streptomyces bacteria beginning in the late 1940s. The tetracyclines act by blocking the binding of aminoacyl tRNA to the A site on the ribosome. However. However. Pseudomonas aeruginosa is less sensitive but is generally susceptible to tetracycline concentrations that are obtainable in the bladder. The tetracyclines are broad-spectrum antibiotics with a wide range of activity against both Gram-positive and Gramnegative bacteria. Tetracycline. chelocardin) have been shown to act by inserting into the bacterial membrane. Thus a blood level of tetracycline which is harmless to animal tissues can halt protein synthesis in invading bacteria. The tetracycline core structure.

but currently it is produced entirely by chemical synthesis. acne and rosacea. aplastic anemia develops in a small proportion (1/50. sinusitis. Chloramphenicol was originally discovered and purified from the fermentation of a Streptomyces species.Doxycycline is a semisynthetic tetracycline developed in the 1960s. Chloramphenicol is a protein synthesis inhibitor that has a broad spectrum of activity but it exerts a bacteriostatic effect. Chemical structure of chloramphenicol Chloramphenicol is entirely selective for 70S ribosomes and does not affect 80S ribosomes. Unfortunately. they are subject to inhibition by some of the protein synthesis inhibitors including chloramphenicol. Its unfortunate toxicity towards the small proportion of patients who receive it is in no way related to its effect on bacterial protein synthesis. it is indicated even for use in children for this illness. Chloramphenicol was once a highly prescribed antibiotic and a number of deaths from anemia occurred before its use was curtailed. Now it is seldom used in human medicine except in life-threatening situations (e. chlamydia. It is effective against intracellular parasites such as the rickettsiae. it is used in the treatment and prophylaxis of anthrax and in prophylaxis against malaria. Such cells include the blood forming cells of the bone marrow. The eucaryotic cells most likely to be inhibited by chloramphenicol are those undergoing rapid multiplication. thereby rapidly synthesizing mitochondria.000) of patients. typhoid fever). since mitochondria originated from procaryotic cells and have 70S ribosomes. syphilis. the inhibition of which could present as aplastic anemia. Chloramphenicol inhibits the bacterial enzyme peptidyl transferase. . It is frequently used to treat chronic prostatitis. In addition. It is also effective against Yersinia pestis (the infectious agent of bubonic plague) and is prescribed for the treatment of Lyme disease. ehrlichiosis and Rocky Mountain spotted fever. thereby preventing the growth of the polypeptide chain during protein synthesis.g. Because doxycycline is one of the few medications that is effective in treating Rocky Mountain spotted fever (with the next best alternative being chloramphenicol). This likely explains the toxicity of chloramphenicol. pelvic inflammatory disease. However.

Macrolides inhibit bacterial protein synthesis by binding to the 50S ribosomal subunit. most often bacteria causing middle ear infections. thus making the lactone ring 15membered. bronchitis. Bacteroides). .g. shown above. throat infections. such as non-gonococcal urethritis and cervicitis. Lincomycin has activity against Gram-positive bacteria and some Gram-negative bacteria (Neisseria. pneumonia and sinusitis. Lincomycin and clindamycin are a miscellaneous group of protein synthesis inhibitors with activity similar to the macrolides. Macrolides are bacteriostatic for most bacteria but are cidal for a few Grampositive bacteria. It is frequently used as a penicillin substitute and is effective against Gram-negative anaerobes (e. Legionella and Haemophilus. Neisseria. Chemical structure of a macrolide antibiotic. Azithromycin is one of the world's bestselling antibiotics. The most important members of the group are erythromycin and oleandomycin. is a subclass of macrolide antibiotics. erythromycin. Binding inhibits elongation of the protein by peptidyl transferase or prevents translocation of the ribosome or both. but not against the Enterobacteriaceae. influenzae). Clindamycin is a derivative of lincomycin with the same range of antimicrobial activity. It is also effective against certain sexually transmitted diseases.The macrolide family of antibiotics is characterized by structures that contain large lactone rings linked through glycoside bonds with amino sugars. laryngitis. tonsillitis. H. but it is considered more effective. but it differs chemically from erythromycin in that a methyl-substituted nitrogen atom is incorporated into the lactone ring. Azithromycin. Erythromycin is active against most Gram-positive bacteria. It is s derived from erythromycin. Azithromycin is used to treat certain bacterial infections.

Nalidixic acid belongs to a group of compounds called quinolones. K. Binding of the drug inhibits DNA gyrase activity. However. which is caused by a Gram-positive bacillus. The most severe common adverse effect of clindamycin is Clostridium difficile-associated diarrhea (the most frequent cause of pseudomembranous colitis). it is classically linked to clindamycin use. . and affect animal cells and bacterial cells alike and therefore have no therapeutic application. The compound is unusual in that it is effective against several types of Gram-negative bacteria such as E. including beta-lactam antibiotics. Two nucleic acid synthesis inhibitors which have selective activity against procaryotes and some medical utility are the quinolones and rifamycins. (ciprofloxacin) was recently touted as the drug of choice for treatment and prophylaxis of anthrax. Either case. The fluoroquinolone. Bacillus anthracis. The majority of these drugs are unselective. however. Although this side-effect occurs with almost all antibiotics. Nalidixic acid is a synthetic chemotherapeutic agent that has activity mainly against Gramnegative bacteria. coli. Effects on Nucleic Acids Some antibiotics and chemotherapeutic agents affect the synthesis of DNA or RNA. or can bind to DNA or RNA so that their messages cannot be read. pneumoniae and Proteus species which are common causes of UTIs. It is a common topical treatment for acne.Clindamycin is a lincosamide antibiotic. such as malaria. of course. It is not usually effective against Pseudomonas aeruginosa. and can be useful against some methicillin-resistant Staphylococcus aureus (MRSA) infections. Cipro. Some quinolones penetrate macrophages and neutrophils better than most antibiotics and are thus useful in treatment of infections caused by intracellular parasites. Enterobacter aerogenes. and Gram-positive bacteria may be resistant. Nalidixic acid is a bactericidal agent that binds to the DNA gyrase enzyme (topoisomerase) which is essential for DNA replication and allows supercoils to be relaxed and reformed. can block the growth of cells. It is usually used to treat infections with anaerobic bacteria but can also be used to treat some protozoal diseases. the main use of nalidixic acid is in treatment of lower urinary tract infections (UTI). Some quinolones have a broadened spectrum against Gram-positive bacteria.

which is an enzyme necessary to separate replicated DNA. Competitive Inhibitors . Neisseria gonorrhoeae. It is effective orally and penetrates the cerebrospinal fluid so it is useful for treatment of bacterial meningitis. Rifampicin is a semisynthetic derivative of rifamycin that is active against Grampositive bacteria (including Mycobacterium tuberculosis) and some Gram-negative bacteria. especially when isoniazid resistance is indicated. It is also used to treat infection by Listeria monocytogenes. and also has a role in the treatment of methicillin-resistant Staphylococcus aureus (MRSA) in combination with fusidic acid. Haemophilus influenzae and Legionella pneumophila. and thereby inhibits cell division. The rifamycins are a comparatively new group of antibiotics. tuberculosis than other anti-tuberculosis drugs. It has been found to have greater bactericidal effect against M. Rifampicin acts quite specifically on the bacterial RNA polymerase and is inactive towards DNA polymerase or RNA polymerase from animal cells. thereby blocking mRNA synthesis. It is a semisynthetic compound derived from Amycolatopsis rifamycinica (formerly known as Amycolatopsis mediterranei and Streptomyces mediterranei). a fluoroquinolone is a broad-spectrum antimicrobial agent that is active against both Gram-positive and Gram-negative bacteria. It functions by inhibiting DNA gyrase. It is used in prophylactic therapy against Neisseria meningitidis (meningococcal) infection. including tuberculosis and leprosy. a type II topoisomerase. The antibiotic binds to the beta subunit of the polymerase and apparently blocks the entry of the first nucleotide which is necessary to activate the polymerase. and it has largely replaced isoniazid as one of the front-line drugs used to treat the disease. Rifampicin (or rifampin) is a bactericidal antibiotic from the rifamycin group.Ciprofloxacin (cipro). Rifampicin is typically used to treat Mycobacterium infections. also the products of Streptomyces species.

Animal cells do not synthesize their own folic acid but obtain it in a preformed fashion as a vitamin. Their selective toxicity is based on the premise that the bacterial pathway does not occur in the host. The sulfonamides (e. Since animals do not make folic acid. the substrate for the first enzyme in the THF pathway. and they competitively inhibit that step. coli. but they do not fulfill their metabolic function in the cell. Sulfonamides are structurally similar to para aminobenzoic acid (PABA). Sulfonamides were introduced as chemotherapeutic agents by Domagk in 1935. they are not affected by these drugs.Many of the synthetic chemotherapeutic agents (synthetic antibiotics) are competitive inhibitors of essential metabolites or growth factors that are needed in bacterial metabolism. coli. competitive inhibitors are structurally similar to a bacterial growth factor or metabolite. The sulfonamides have been extremely useful in the treatment of uncomplicated UTI caused by E. Bacteria which are almost always sensitive to the sulfonamides include Streptococcus pneumoniae. Trimethoprim is structurally similar to dihydrofolate (DHF) and competitively inhibits the second step in THF synthesis mediated by the DHF reductase. Hence. Gantrisin and Trimethoprim) are inhibitors of the bacterial enzymes required for the synthesis of tetrahydofolic acid (THF). who showed that one of these compounds (prontosil) had the effect of curing mice with infections caused by beta-hemolytic streptococci. At a chemical level. since they are designed to specifically inhibit an essential metabolic pathway in the bacterial pathogen. these types of antimicrobial agents are sometimes referred to as anti-metabolites or growth factor analogs. which achieve their selective toxicity for bacteria . Some are bacteriostatic and some are bactericidal. but differ widely in their pharmacological actions. and in the treatment of meningococcal meningitis (because they cross the blood-brain barrier). The resulting sulfonamides have broadly similar antibacterial activity. Chemical modifications of the compound sulfanilamide gave rise to compounds with even higher and broader antibacterial activity. betahemolytic streptococci and E.g. the vitamin form of folic acid essential for 1-carbon transfer reactions.

Three additional synthetic chemotherapeutic agents have been used in the treatment of tuberculosis: isoniazid (INH). Hence. DHPS. Since the tubercle bacillus rapidly develops resistance to the antibiotic. a key step in folate synthesis. para-aminosalicylic acid (PAS). but now. cells will be unable to divide. similar in activity to the sulfonamides. but now it is a secondary agent. The chemical structures of sulfanilamide and para-aminobenzoic acid (PABA). Ethambutol inhibits incorporation of mycolic acids into the mycobacterial cell wall. rifampicin is given) in conjunction with INH and ethambutol. . Isoniazid has been reported to inhibit mycolic acid synthesis in mycobacteria and since it is an analog of pyridoxine (Vitamin B6) it may inhibit pyridoxine-catalyzed reactions as well. It must also be pointed out that the tubercle bacillus rapidly develops resistance to ethambutol and INH if either drug is used alone. ethambutol and INH are given to prevent outgrowth of a resistant strain. Isoniazid is activated by a mycobacterial peroxidase enzyme and destroys several targets in the cell. sulfanilamide and other sulfonamides exhibit a bacteriostatic rather than bactericidal effect. having been largely replaced by ethambutol. PAS is an anti-folate. The usual strategy in the treatment of tuberculosis has been to administer a single antibiotic (historically streptomycin. Folate is necessary for the cell to synthesize nucleic acids (DNA and RNA).on this basis. PAS was once a primary anti-tuberculosis drug. and ethambutol. and in its absence. most commonly. sulfanilamide acts as a competitive inhibitor of the enzyme dihydropteroate synthetase. In bacteria. which catalyses the conversion of PABA to dihydropteroate.

Isoniazid is never used on its own to treat active tuberculosis because resistance quickly develops. . Isoniazid is a first-line anti-tuberculosis medication used in the prevention and treatment of tuberculosis.Isoniazid is also called isonicotinyl hydrazine or INH.