Tetracycline

PHRM 304

The tetracyclines were the first major group of antimicrobial agents for which the term broad spectrum was used- namely, they exhibit activity against both gram-positive and gram-negative bacteria. Although they bind both the eukaryotic 80S, and the bacterial 70S units of Ribosome, the latter is more sensitive, making tetracyclines effective and selective antimicrobial agents. Contraindicated to young and pregnant as it form chelate with calcium and thus causes problem in development of bones.

Tetracyclines enter through porins in Gram-negative bacteria, and through their lipophilicity in Grampositive bacteria. They pass through the cytoplasmic membrane via active transport, where they are mistaken for food. Available agents:

Tetracycline

Minocycline

Doxycycline

Antibacterial synthesis:

agents

which

impair

protein

Inhibition of protein synthesis by an effect on ribosome: Drugs acting on 30S subunits: - Aminoglycoside - Tetracyclines Drugs acting on 50S subunit: - Chloramphericol - Erythromycin

Ribosomes are complexes of RNA and protein that are found in all cells. The ribosome functions in the expression of the genetic code from nucleic acid into protein, in a process called translation. Ribosomes do this by catalyzing the assembly of individual amino acids into polypeptide chains; this involves binding a messenger RNA (mRNA) and then using this as a template to join together the correct sequence of amino acids. The ribosomal subunits of prokaryotes and eukaryotes are quite similar. The unit of measurement is the Svedberg unit, a measure of the rate of sedimentation in centrifugation rather than size and accounts for why fragment names do not add up.

Prokaryotes (are a group of organisms that lack a cell nucleus) have 70S ribosomes, each consisting of a small (30S) and a large (50S) subunit. Eukaryotes have 80S ribosomes, each consisting of a small (40S) and large (60S) subunit.

Tetracycline
Tetracyclines are named for their four (tetra-) hydrocarbon rings (-cycl-) derivation (-ine).

Mode of action of tetracycline

The tetracyclines of clinical importance interfere with protein synthesis at the ribosomal level. They bind to 30S subunit of the bacterial ribosome. Tetracycline antibiotics inhibit protein synthesis by inhibiting the binding of aminoacyl-tRNA to the mRNAribosome complex. Tetracycline inhibits cell growth by inhibiting translation. The binding is reversible in nature.

Drugs enter bacteria in part by passive diffusion, in part by an energy-dependent process of active transport, result in susceptible cells concentrate the drug, the intracellular drug concentration is much higher than the extracelluar one. Once inside the cell, tetracyclines bind reversibly to receptors on the 30S subunit of the bacterial ribosome in a position that blocks the binding of the aminoacyl-tRNA to the accepter site on the mRNAribosome complex.

Antibacterial activities
The tetracyclines as a whole have a broad spectrum of activity and are the most widely prescribed after penicillins. Bacteriostatic for many gram-positive and gramnegative bacteria. Bacteriostatic for chlamydiae, spirochetes amoebae). mycoplasma, rectettsia, & some protozoa (e.g.

Structure: Tetracycline

General characteristics of tetracyclines

Four rings present in the tetracycline nucleus. The tetracyclines are amphoteric compounds, i.e., forming salts with either acids or bases. In neutral solutions these substances exist mainly as Zwitterion. The acid salts of the tetracyclines are formed through protonation of the dimethylamino group of C4. The corresponding hydrochloride salts are used most commonly for oral administration and are usually encapsulated owing to their bitter taste.

 The tetracyclines form stable chelate complexes with many metals, e.g., Ca++, Mg++, Fe++, etc. Chelation is an important feature of the chemical and clinical properties tetracyclines. The acidic functions of the tetracyclines are capable of forming salts through forming chelation with metal ions. Deposition of tetracyclines in the bone and teeth occurs during calcification in growing children, which causes discoloration and hypoplasia of the teeth.

 Tetracyclines attack protein synthesis in mammalian cells as well as in bacterial cells. Fortunately, bacterial cells accumulate the drug far more efficiently than mammalian cells and are therefore more susceptible. Unfortunately, it does have side-effects due to the fact that it kills the intestinal flora (such as Escherichia coli) that make vitamin K (mainly vitamin K2)- a vitamin which is needed as part of the clotting process. Thus tetracyclines delay blood coagulation process.

Commercially available tetracyclines

First Generation (Dose Chlortetracycline Oxytetracycline Tetracycline Demeclocycline Second Generation (Dose interval longer) Minocycline Methacycline Doxycycline Third Generation (Derivative of Second Gen) Glycylcycline (Derivative of Minocycline) intervals shorter)

Structure: Tetracycline

SAR of tetracyclines
A. Linear arrangement of four rings is essential for activity. Derivatives having fewer than 4 rings are inactive. B. Opening of rings or addition of ring produces inactive compounds. Tetracyclines contain four fused rings (A-D), one of which, D, is aromatic; rings A and B contain sites of unsaturation. Essentially all alterations to the general tetracycline ring skeleton are deleterious; breaking any ring, disrupting aromaticity in ring D, or aromatization of ring A or C destroys all tetracycline activity.

SAR of tetracyclines
C. The presence of 4-dimethylamino is essential for activity and for optimum distribution. Removal of the 4-dimethylamino group affords a loss of about 75% of the antibiotic effect of the parent tetracyclines. Removal of the dimethylamino group at C4 diminishes in vitro activity and abolishes in vivo activity.

D. Modifications to the functional groups on ring A and the bottoms of rings B, C, and D are generally not tolerated, but extensive modification is possible elsewhere. From C-5 to C-9 can be altered in various ways: a. Removal of 5-OH (present in oxytetracycline) to give 5-deoxycompounds does not change the activity. b. Neither 6-methyl nor 6-hydroxy is essential. Neither the hydroxyl nor the methyl at C6 is essential for activity; no direct ribosome interaction is observed for the C6 substituents. c. Electron withdrawing groups (-Cl, -NO2, protonated dimethyl amino group) at 7 enhance the activity.

10

E. The conjugation between C-10 and C-12 is essential for activity. F. In the 2-amide moiety, one amide hydrogen can be replaced without loss of activity.

Tetracycline side effects

The side effects of tetracyclines are as a result of imperfections in distinguishing the 70S and 80S subunits. This is what causes liver and kidney damage at high doses or toxicity during pregnancy. Tetracyclines are teratogens due to the likelihood of causing teeth discoloration in the fetus as they develop in infancy. For this same reason, tetracyclines are contraindicated for use in children under 12 years of age. They are however safe to use in the first 18 weeks of pregnancy.

Tetracyclines were the first broad-spectrum antibiotics and have been used successfully for decades to treat both gram-positive and gram negative bacterial infections. Chlortetracycline was the first tetracycline to be isolated, in 1948, from Streptomyces aureofaciens. Biochemical probing identified multiple tetracycline binding sites within 30S, one of which resides in the decoding A-site and seems to be responsible for the antibiotic effect of tetracycline. It has been accepted for some time that tetracycline interferes with proper tRNA binding to the A-site.

Molecular interaction between Tetracycline and 30S

How many asymmetric carbons does tetracycline have?

First, locate all the SP3 hybridized carbons in tetracycline. Only SP3 hybridized carbons can be asymmetric carbons, because an asymmetric carbon must have four different groups attached to it. Tetracycline has nine hybridized carbons.

How many asymmetric carbons does tetracycline have?

Four of them (#1, #2, #5, and #8) are not asymmetric carbons because they are not bonded to four different groups. Tetracycline, therefore, has five asymmetric carbons.