TetracyclinesAre bacteriostatic antibiotics having broad

spectrum of activity.Isolated from Streptomyces bacteria.First one isolated was chlortetracycline

(1948).

They inhibit protein biosynthesis by binding to 30s ribosomal subunit and prevent aminoacyl tRNA from binding to the A-site.

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OOOH

Cl

OH

OH

O

NH2

OH

NHHHO

4a5a

11a 12a

Chlortetracycline

Mechanism of actionTetracyclines could inhibit protein synthesis in

human, but they normally can not penetrate the mammalian cell membrane.

The transport of tetracycline into the cell (especially the gram –ve bacteria) needs:1. a passive diffusion through porines, this process is

pH dependent and required proton-driven carrier protein. This protein is only present in bacteria not in human cell.

2. Active transport: requires Mg++ and ATP.This is why tetracyclines are quite selective on the

bacterial cell.

Clinical uses of tetracyclinesThey have the broadest spectrum of activity, on both

gram +ve, gram –ve and atypical bacteria.Resistance has been developed rapidly against

tetracyclines, as a result of that penicillins have replaced them in many infections, especially the respiratory infections.

Tetracyclines are still used in rickettsia, Chlamydia, mycoplasma and acne infections.

Some of them have antiparasitic properties such as the use of Doxycycline in the treatment and prophylaxis of malaria.

They have bacteriostatic action, not recommended in life threatening infections such as septicemia, endocarditis and meningitis

Clinical uses of tetracyclinesTetracyclines should be avoided in children and

pregnant women: they bind to the growing teeth and bones…. Lead to tooth discolorations and toxicity in fetus.

Tetracyclines can be divided according to the duration of action into:1. Short acting: chlortetracycline (t1/2 = 7hr).

2. Intermediate acting: tetracycline and demeclocycline (t1/2 10-15hr).

3. Long acting: Doxycycline and minocycline (t1/2 = > 16hrs)

Tetracycline chemical structureThey are derivatives of

octahydronaphthacene which comprise four fused six-membered rings.

The structure have 5 or 6 chiral centres.They have acidic and basic characteristics.

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OOOH

Cl

OH

OH

O

NH2

OH

NHHHO

4a5a

11a 12aABCD

pKa = 9.5

pKa = 7.0

pKa = 3

Chemical instabilityThey have the ability to epimerize at acidic conditions

upon long standing:Epimers are diastereomers that differ in the

configuration at one stereogenic center.

The epitetracycline is much less active compared to the natural isomer.

For this reason tetracyclines should be freshly prepared and used to gain the desired maximum activity.

O

NH2

O

OH

NH

O

NH2

O

OH

NH

Natural form Epitetracycline

Chemical instabilityAcid instability: in presence of acidic conditions, they undergo dehydration at C5,6. this followed by double bond migration from C11a,12 to C11,11a.

OOOH OHOH

O

NH2

OH

NHHHO

H+

OOOH OHOH

O

NH2

OH

NH

spontanuous rearrangement

OOHOH OOH

O

NH2

OH

NH

5,6-anhydrotetracycline (inactive)

Chemical instabilityBasic instability: Bases promotes a reaction between C6 and C11a to form the inactive isotetracycline with a lactone ring.Derivatives have been synthesized with the 6-OH group been removed, these agents were more stable and long lasting than those with 6-OH group.

OOOH OHOH

O

NH2

OH

NHHHO

OH-

OOOH OOH

O

NH2

OH

NH

O

Isotetracycline (inactive)

Chemical instabilityTetracyclines can form stable metal chelates

especially with dicationic such as Ca++ and Mg++ and Fe++.

The chelates are highly water insoluble…. Not orally available.

For this, tetracyclines should not be given along with milk, antacids, or iron salts.

Another consequence of this is the high affinity of tetracycline to chelate with bone calcium… deposited in bone and teeth:Affect bone growth.Tooth discoloration.

SAR in tetracyclinesDerivatives with less than four fused rings were inactive.The simplest structure with retained activity

was 6-demethyl-6-deoxytetracycline.Substituents at C1,2,3,4,10,11,11a,12 can not

be modified for better activity.Slight modifications on ring A found tolerable

without dramatic loss in activity.

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OOOH

R1

OHOH

O

NH2

OH

NHH

ABCD

R4R2 R3

SAR in tetracyclines Regarding Ring A:

The enolized carbonyl system between C1 and C3 is essential for activity and can not

be modified especially the amide group:1. Replacement of the amide with nitro or aldehyde

abolished the activity.2. Monoalkylation of the amide reduced the activity.

Dimethylamino at C4 can be free amine or N-methylamino, but can not accept larger alkyl than methyl.

Dimethylamino must have an α-orientation, the other isomer is much less active.

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OOOH

R1

OHOH

O

NH2

OH

NHH

ABCD

R4R2 R3

SAR in tetracyclines Ring A and B should have cis- fusion with OH at C12a. OH group at C12a must be free,

esterification abolished the activity. Hydrophobic substitution at C5,5a,6,7,8,9

resulted in retention and sometimes improvement in activity.

The presence of 5-OH does not have important role in activity (such as in demeclocycline and oxytetratcycline compared to minocycline and doxycycline)

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OOOH

R1

OHOH

O

NH2

OH

NHH

ABCD

R4R2 R3

SAR in tetracyclines Acid stable scaffold (6-deoxy or 6-demethyl-6-deoxytetracyclines) were used to prepare derivatives with mono

and disubstitutions at C7 (mainly) and C9… either EDG or EWD:

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OOOH

R1

OHOH

O

NH2

OH

NHH

ABCD

R4R2 R3

OOOH OHOH

O

NH2

OH

NHH

N

Minocycline

OOOH OHOH

O

NH2

OH

NHH

Cl OH

Demeclocycline

SAR in tetracyclines Neither 6α nor 6β-OH is essential for activity (Doxycycline and methacycline

are more active than oxytetracycline). These derivatives are also more stable toward

acid and base inactivation. More lipid soluble… better absorbed orally (>90% orally available). High protein binding… have long duration of action.

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OOOH

R1

OHOH

O

NH2

OH

NHH

ABCD

R4R2 R3

OOOH OHOH

O

NH2

OH

NHH

Methacyclinet1/2 = 12 hrsmore potent than 6-oxytetracycline

OH

Metabolic transformations in tetracyclinesMost of them are excreted unchanged in

urine.Sulfate and glucuronide conjugates were

detected in urine especially for Doxycycline and minocycline.

The major metabolite found to be the N-dealkylated at C4, and to a little extent at C7 (for minocycline).

OOOH OHOH

O

NH2

OH

NHH

N

Sulfate conjugationglucuronidation

N-dealkylation

Glucuronidation

Tetracycline new generations The main goal of recent research in

tetracyclines is to discover agents that might be effective against resistant strains.

The target ring for structural modification is ring D, at C7,8 and 9 (Glycylcylines).

Some derivatives were synthesized by adding substituent at the 2-amido group (Rolitetracycline)

GlycylcylinesHave broad spectrum of activity.More active than old tetracyclines against the

resistant strains.Tigecycline:

Long t1/2 of 55.4 hrs.Highly bound to plasma protein.Mainly used as IV for skin infections and intra-abdominal infections.Active against MRSA and S. pneumoniae (resistant to penicillin)

OOOH OHOH

O

NH2

OH

NHH

N

NH

O

N

Glycylcyline

OOOH OHOH

O

NH2

OH

NHH

N

NH

OHN

Tigecycline

Rolitetracycline

is a semi-synthetic tetracycline (pyrrolidinylmethyltetracycline).

Has broad-spectrum activity used especially for parenteral administration.

Mainly used in rickettsia and brucellosis.Recent studies trying to investigate the effect

of its combination with group of β-lactams such as cefotaxime for treating MRSA infections.

OOOH OHOH

O

HN

OH

NHH

N

OH