‫معهد اإلسراء لمعادلة الشهادة‬

ANTI-BIOTICS
Chemical substance produced by M.O (Micro-organism) that has the capacity, in low concentration to
inhibit selectively or even to destroy bacteria and other M.O. through antimetabolic mechanism.

M.O. producing A.D. called (Actinomycetes)

In searching for new A.B. , relatively simple and rapid methods have seen develeped for screening M.O. ,
for A.B. producing ability soil sample are commonly employed in the screen because they are a rich source
of A.B. producing organisms. A general method for screening first involves treating the soil sample with
chemical that inhibit the growth of interfering bacteria and fungi but do not affect actinomycetes,
cycloheximide is an as antifungal often employed for this purpose and 1 dilution of phenol is used as anti-
bacterial agent.

Varying dilution of the treated soil sample are streaked on agar plates containing medium that supports the
growth of actinomycetes. After incubation for 3-7 days at25-30C • the plates are examined for
characteristic colonies of actinomycetes, these colonies then transferred on to fresh medium contain
pathogenic M.O. for indication of the potential usefulness of the A,B. for example activity against G+ve
bacteria can be determined with Staphylococcus aureus or Bacillus subtilis, activity against G-ve bacteria
can be determined with E. coil or Salmonella typhi and antifungal with Neurospora crassa .

The next step in the screening is to determine whether the chemical substance that produced the inhibition
is a new A.B. or a known compound , a rapid method that has been developed for this determination is
termed (Bio auto-graphy assay)

 This assay employs paper or thin layer chromatography TLC and biologic assay. Extract containing
the newly discovered A-B. is chromatographed along with reference in different solvent systems .
Because each A.B. would possess a characteristic mobility on the chromatogram in a given solvent
system, a comparison of the mobilities of the unknown A.B. with those of known one in several
solvent system would indicate whether the newly discovered A.B. was a known compound.

 The detection methods of the A.B. on the developed chromatogram :

Chemical methods for detection is impossible and difficult because the A.B. are widely diverse chemically,
consequently a biologic method is used to detect the AB. by placing the developed chromatogram on an
agar medium that has been seeded with an appropriate test organism, the anti-biotics diffuse from the
chromatogram in to the agar and after incubation , clear zone on the agar owing to inhibition of growth of
the test organism indicate the position of the AB. on the chromatogram.

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 Commercial production of anti-biotics

When a new A.B. has been discovered , investigation in to the chemical, physical, biologic properties of
the A-B are required , the most important requirement for commercial production of A.B. is:

 The organism must excrete the A.B. in to culture medium, however some anti-biotics such as
those of the polyene group are retained in the cells of organism and required special extraction
procedures for recovery which is very difficult and expensive ‫👈مهم‬

 Phases of anti-biotics production:

 In the production of A.B. there are two distinct phases in the fermentation process:
 The growth phase of the organism which is termed (Tropho phase) & anti-biotics
To production phase ( Idio phase)
 Example: in a course of typical penicillin fermentation carried out in a culture medium containing
glucose and lactose as a sources of carbon nutrition , corn steep liquor for nitrogen sources and
phosphate buffer. ‫👈 مهم‬

 During the growth phase, the culture becomes thick owing to the formation of aggregates of
fungal cells called ( Mycelium) , so the first phase ( tropho phase) last from the beginning of the
culture period to approximately one day later ( 0-24 hr) , during the growth phase glucose rather
than lactose is preferentially utilized because it can be used directly as a source of carbon. In the
growth phase process, ammonia is liberated by deamination of amino acid of the com steep liquor,
this liberation raise the PH of the medium to 7 which is the optimum PH for penicillin stability and
buffer in the medium maintain the PH close to neutrality.

 Penicillin production increase rapidly between 24-80 hr , at the start of A.m production phase
glucose has been used up and the fungus then uses lactose for carbon source but lactose cannot
be utilized until its hydrolyzed to glucose and galactose so:

 The decreased availability of carbon source is thought to be the triggering mechanism for penicillin
production ‫👈 مهم‬

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 Factors that are often observed to have qualitative and quantitative

importance for A.B. production:

 Sources of nutritional carbon and nitrogen Ratio of carbon to nitrogen nutrients (C/N)

 Mineral composition of medium

 Incubation temperature

 Initial PH & control of PH during the fermentation period

 Rate & method of aeration

 Addition & timing of addition of special growth and A.B. promoting

substance

For example : some strain of Bacillus yield Bacitracin when C/N ratio is 15 , at lower ratio the yield
is less& when the ratio is reduced to 6 another undesired A.B. is produced.

Another example : the use of mercaptothiazole in the culture of streptomyces which give
tetracycline will favor chlortetracycline.

 Classification of A.B.

1. According to the general mode of A.B. action:

inhibition of protein synthesis example : chloramphenicol, clindamycin, erythromycin, gentamycin,

lincomycin, neomycin, tetracycline, streptomycin.
alteration in cellular membrane function exp. Amphotericin, nystatin, polymyxin.
inhibition of cell wall formation : penicillin , cephalosporin, vancomycin, bacitracin, cycloserine
disruption of deoxy-ribonucleic acid metabolism : exp. Actinomycin , doxorubicin, mitomycin,
rifampin, bleomycin, novobiocin.

2. According to the biosynthetic pathways:

 A.B. derived from amino acid metabolism

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 A.B. derived from acetate metabolism

 A.B. derived from CHO metabolism

A.B. derived from amino acids

Penicillin

Penicillin is a group of antibiotics derived from Penicillium fungi.They include penicillin G, procaine
penicillin, benzathine penicillin, and penicillin V. Penicillin antibiotics are historically significant
because they are the first drugs that were effective against many previously serious diseases, such
as syphilis, and infections caused by staphylococci and streptococci. Penicillins are still widely used
today, though many types of bacteria are now resistant.
 All penicillins are ß-lactam antibiotics and are used in the treatment of bacterial infections caused
by susceptible, usually Gram-positive, organisms

🙄.Beta-lactam ring and Thiazolidine ring ‫مهم‬

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 Biosynthesis

There are three main and important steps in the biosynthesis of penicillin G (benzylpenicillin).

 The first step is the condensation of three amino acids :

 amino-adipic acid, L-cysteine, L-valine into a tripeptide🙁 Before condensing into the
tripeptide, the amino acid L-valine must undergo epimerization to become D-valine. The
condensed tripeptide is named -(L-aaminoadipyl)-L-cysteine-D-vaIine (ACV).

 Second step in the biosynthesis of penicillin G is the oxidative conversion of linear ACV
into the bicyclic intermediate isopenicillin N by isopenicillin N synthase (IPNS),
 Isopenicillin N is a very weak intermediate, because it does not show strong antibiotic
activity

 The final step is a transamidation by isopenicillin N Nacyltransferase, in which the a-

aminoadipyl side-chain of isopenicillin N is removed and exchanged for a phenylacetyl
sidechain.
 This reaction is encoded by the gene penDE, which is unique in the process of obtaining
penicillins.[ysteine-D-vaIine synthetase (ACVS)

So…

 First ➡️(ACV).
 Second ➡️(IPNS)
 Final ➡️(ACVS)

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ß-lactam ring of penicillin is susceptible to cleavage by a variety of reagents as well as some enzymes.

The labile "-lactam ring of penicillins

and other b-lactam antibiotics is characterized by its pronounced susceptibility to various nucleophiles,
acid-base reagents, metal ions, oxidizing agents.

The instability of Penicillin in acidic & alkaline media observed to be a major hurdle in the development of
penicillin and other useful b-lactam antibiotics. Therefore, degradation or stability study of b-lactam
antibiotics has been of paramount importance not only for their market availability, but also to evaluate
their pharmacokinetic properties and adverse reactions.

‫👈انتبه‬

 Penicilloic acid

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 Penicillic acid

 Aminobenzyl p. (ampicillin)
acid-stable but inactivated by penicillinase ez

 p-hydroxy aminobenzyl penicillin(AmoxiciIlin)

acid-stable but inactivated by penicillinase ez.

 Carbenicillin: carboxybenzyl-penicillin:
extended spectrum against G-ve Bacteria. Specially Pseu. & E. coli

 Cloxacillin, dicloxacillin, oxacillin, methicillin & nafcillin :

 are semisynthetic P. that are not inactivated by penicillinas ez. ‫👈👈 انتبه‬

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Clavulanic acid: is a fermentation product of Streptomyces clavuligerus , structurally related to p.

very weak A.B. but is capable of irreversible inactivating bacterial ß- lactamase responsible for A.B.
resistance ------------ Synergistic action

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 cephalothin is a first-generation cephalosporin antibiotic.

 Cefoxitin is second—generation Its activity spectrum includes a broad range of gram-negative
and gram-positive bacteria including anaerobes. It is inactive in vitro to most strains of
Pseudomonas aeruginosa and many strains Enterobacter
 Cefotaxime
is a third-generation cephalosporin antibiotic. Like other third-generation cephalosporins, it has
broad spectrum activity against Gram positive and Gram negative bacteria.

 Cefepime is a fourth-generation cephalosporin antibiotic. Cefepime has an extended spectrum of

activity against Gram-positive and Gramne gative bacteria, with greater activity against both
types of organism than third-generation agents.

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Chloramphenicol

 Steps involved in the synthesis of chloramphenicol

1- Trans-amination + first step
2- Hydroxylation
3- Acylation
4- Reduction of carbox group (COOH) R-OH JS
5- Terminal oxidation of Anuno group final step

 obtained from culture of Streptomyces venezuelae,its derived biosynthetically from

a.a. phenyl alanine / or tyrosine.

 P-aminophenylpyruvic acid has been suggested as an early metabolite in the pathway &
subsequent steps involving transamination, hydroxylation, acylation, reduction of carboxyl
group and terminal oxidation of the amino group are suspected

 Chloramphenicol act by inhibiting protein synthesis at the ribosome level. However ,

occasional development of a plastic anemia and related blood dyscrasias which may be irreversible
and fatal introduces a serious limitation on the therapeutic utility of this anti-biotic, although
chloramphenicol may still the drug of choice for acute typhoid fever and other sever Salmonella
infection

 Chloramphenicol is stable but esters of the A.B. are employed in certain pharmaceutics formulation
for solubility purposes. These esters are hydrolyzed in the body to release the physiologically active
molecule. The insoluble palmitate ester is used in some oral formulation to avoid the bitter taste of
A.B. , and the mono-sodium succinate ester is used for greater water solubility in the preparation
for intra-venous use. ‫👈👈 مهم‬

1. Cephalothin

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2. Cefoxitin

 B-lactam ring , Di-hydrothiazine ring and Thiophene ring

1- 3rd Cefotaxime
2- 4th Cefepime
 B-lactam ring ,Di-hydrothiazine ring and Thiazole ring

1- Oxacillin
2- Cloxacillin
3- Dicloxacillin
 Thiazolidine ring,B-lactan ring and Oxazole ring

A.B derived trom acetate metabolism

Tetracycline:

 -a broad-spectrum antibiotics obtained from Streptomyces aureofaciens

 -It is commonly used to treat acne' infection, and other infections caused by bacteria.
 -The first of these compounds was chlortetracycline followed by oxytetracycline and
tetracycline.

 Tetracycline divided into:➡️

 Naturally occurring:

1 -tetracycline 2-chlortetracycline

3-oxytetracycline 4-demeclocycline

 Semisynthetic occurring:

1 -doxycycline 2-minocycline

3-meclocycline 4-lymecycline

5-methacycline 6-rolitetracycline

 Total Synthesis of the Tetracyclines:

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8 malonate of acetate units condensation + addition of C2+ decarboxylation give us

polyketide consist of 19 C atomatom‫ه‬

 Structure activity of tetracycline

 The amide hydrogen may be replaced with a methyl group,

but larger groups have a deleterious effect except for those which are elilminated
spontaneously in water .
 The dimethyl amino group may be replaced by a primary amino group without loss of in vitro
activity but all other changes so far lead to decreased bacteriostatic action

 Dehydrogenation to form a double bond between C-5a and C-1 la markedly decreases activity

 Polar substituents at C-5 and C-6 contribute decreased lipid versus water solubility to the
tetracycline

 The drugs are amphoteric,🙂 meaning they will form salts with both strong acids and bases.
Thus, they may exist as salts of sodium or chloride.

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Total Synthesis of Tetracycline

 Carbonyl-methylene condensations yield: the tetracyclic pretetramide nucleus.

 Methylation of the c-6 position of the pretetramide is an early step in the

biosynthesis of most tetracyclines, but this step is omitted in tie formation of the
naturally occurring de-methyl-tetracyclines
 Hydroxylation of the c-4 position and dearomatization to yield:
4-keto intermediate appears to precede 7-chlorination
Halogenation must precede introduction of the 4-amino group
 Terminal reactions in the biosynthetic sequence are hydroxylation at c-6

Spectrum:

a. The tetracyclines are broad-spectrum antibiotics.

2- They are active against the following microorganisms:

3- gram-positive and gram-negative bacteria

4- spirochetes

5- mycoplasmas,

6- rickettsiae,

7- Candida albicans

 Chlortetracycline:

Uses:

-used as antibacterial and antiprotozoal agent.

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 -Oxytetracycline:

Uses:

 -treat Spirochaetal infection.

 -treating Non-Specific-Urethritis.
 -treating Clostridial wound infection and Anthrax.

Side effects:

1- Local irritation after intramuscular injection

2- Gastrointestinal:-anorexia, nausea, vomiting.
3- Renal toxicity.
4- Hypersensitivity reactions: Urticaria
5- -Blood: Hemolytic anemia, thrombocytopenia, neutropenia

 -tetracycline:

Uses:

1- Tetracycline's primary use is for the treatment of acne vulqaris and rosacea.
2- -It is also used to treat a very wide range of infections.

 Side effects:
1- Gastrointestinal: anorexia, nausea, vomiting, diarrhea'
2- -Skin: rashes, dermatitis.
3- -Renal Toxicity
4- -Hepatic Cholestasis
5- -Hypersensitivity
6- Reactions:Anaphylaxi5
7- Miscellaneous: Dizziness and headache

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 Minocycline:

Uses:

1. typhus fever and Q fever.

2. -Psittacosis Trachoma
3. -Nongonococcal
4. urethritis.
5. -Brucellosis.

 -doxycycline

Uses:

1- used in the treatment of chronic adult periodontitis.

 Other A.B. derived from acetate pathway

Griseofulvin is an antifungal drug that is administered orally. It is used to treat fungal infections of the
skin (commonly known as ringworm) and nails. It is produced by culture of some strains of the mold

Penicillium griseofulvum, from which it was isolated in 1939

The drug binds to tubulin, interfering with microtubule function, thus inhibiting mitosis.

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 Tetracycline is derived from acetate pathway we can say that tetracycline is the major drug
and from it we can drive different derivatives due to limitation in the use of tetracycline.
 Tetracycline is the drug of choice for acne infection.
 Tetracycline and chloro-tetracycline are natural and they are the most drugs that arise from micro-
organism.

 The four functional group which are the Hydroxyl and Carbonyl (came from acetate ) all of them
is carbonyl in origin so they undergo oxidation reduction reaction that will give 2 hydroxyl
group in the structure of tetracycline.

 The amide group at carbon No.2 is very important for the activity of the drug ( any change in it
will lead to loss the activity of the drug ).
 Amide group always take No.2 in numbering .
 7-chlortetracycline is the chemical name of chlortetracycline

Examples

about preparation of the tetracycline derivative:

1. Q\Prepare demeclocycline from tetracycline ?

 Demeclocycline chemical name is (6demethyl-7-chlortetracycline )

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Ans\ tetracycline undergoes demethylation at carbon No.6 and chlorination at carbon No.7

2. Q\Prepare doxycycline from tetracycline ?

 Doxycycline chemical name is (5-hydroxy-6deoxy,tetracycline).

Ans\ Tetracycline undergoes hydroxylation at carbon No.5 and dehydroxylation at carbon No.6.

3. Q\ Prepare minocycline from chlortetracycline?

 Minocycline chemical name is ( 6-deoxy-6demethyl-7-dimethyl amino tetracycline ).

Ans\ Chlortetracycline undergoes dechlorination at carbon No.7 , demethylation and dehydroxylation at

carbon No.6 , amination at carbon No. 7 and dimethylation on amine group at carbon No. 7.

Q\ Prepare minocycline from 7-nitro tetracycline?

Ans\ 7-nitro tetracycline undergoes reduction of the amine group at carbon No. 7 then dimethylation at
the amine group at carbon No. 7 and dehydroxylation and demethylation at carbon No.6.

Homework\ Prepare 5 — hydroxyl -6- deoxy -6methylene -7-nitro from chlortetracycline ?

Ans\Chlortetracycline undergoes hydroxylation at carbon No.5 , dehydration at carbon No.6 dechlorination

at carbon No.7 then amination at carbon No.7 and oxidation of the amine group at carbon No. 7.

A.B. derived from Carbohydrate metabolism

CHO provide the basic metabolic substance for the formation of essentially all microbial products but this
category of A.B. substances is restricted to compounds that are derived directly from CHO.

 These A.B. include amino-glycoside A.B. (Like streptomycin, gentamicin, amikacin,

neomycin & others), and Macrolides
 so aminoglycoside A.B defined as: any antibiotic containing amino-modified sugars
originally isolated from various Streptomyces and Micromonospora species, they are bind
to the16S RNA of the bacterial 30S ribosomal subunit, inhibitin translation and
proteinsynthesis
 . Aminoglycoside use is associated with ototoxicity, neurotxicity and nephrotoxicity.;
they inhibit protein synthesis by binding with the 30S ribosomal subunit. The chemical &
biological properties of these A.B. are similar.
 Common chemical properties include:
water solubility, strong basic character and stability.

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 These A.B. have uncommon sugars linked glycosidically to an amino-substituted aglycone.

 These A.B. have a wide spectrum of activity including many G+ve & G-ve bacteria,
 these A.B. are not absorbed following oral administration & their systemic use is limited by
nephro & oto-toxicities especially with gentamicin & streptomycin, symptoms include
nausea, vertigo, vomiting but recovery is usually complete when therapy is discontinued.
Damage of the auditory branch results in irreversible loss of hearing, auditory toxicity
appears to be more common with amikacin, kanamycin & neomycin.

Streptomycin

Streptomycin is an antibiotic (antimycobacterial) drug, of a class of aminoglycosides and it was the first
effective treatment for tuberculosis. It is isolated from the Streptomvces griseous. Streptomycin is a
bactericidal antibiotic Adverse effects of this medicine are ototoxicity, nephrotoxicity, fetal auditory
toxicity, and neuromuscular paralysis.

 Biosynthesis of Streptomycin

Streptidine and the disaccharide streptobiosamine,which contains the sugar residues,

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2-deoxy-2-methylamino(all these 3 components form the basic structure for streptomycin ) & all these
components are derived from D-glucose as follows :

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Gentamicin

 A broad-spectrum aminoglycoside antibiotic produced by fermentation of

Micromonospora purpurea or M. echinospora.
Gentamicin is an antibiotic complex consisting of three major components (garosamine,
purpurosamine, 2-deoxy streptamine) .
This agent irreversibly binds to the bacterial 30S ribosomal subunit. Specifically, this antibiotic is
lodged between 16S rRNA and S12 protein within the 30S subunit.
This leads to interference with translational initiation complex, misreading of mRNA, thereby
hampering protein synthesis and resulting in bactericidal effect.
 Aminoglycosides are mostly ineffective against anaerobic bacteria, and viruses.👈‫مهم‬

This A.B.derived from CHO by condensation of 2-amino sugar (garosamine &

purpurosamine A) with 2-deoxy streptamine which is also obtained from glucose by
transamination.

Macrolides

Macrolides are a class of natural products that consist of a large macrocyclic lactone
ring to which one or more deoxy sugars, usually cladinose and desosamine, may be
attached.
The lactone rings are usually 14-, 15-, or 16-membered. Macrolides belong to the
polyketide class of natural products.
 Some macrolides have antibiotic or antifungal activity and are used as pharmaceutical
drugs.
Macrolides belong to one of the most commonly used families of clinically important
antibiotics used to treat infections caused by Gram-positive bacteria such as
Staphylococcus aureus, Streptococcus pneumoniae and Streptococcus pyogenes.
Chemically, macrolides are represented by a 14-, 15- or 16-membered lactone ring
carrying one or more sugar moieties and additional substitutions linked to various atoms of
the lactone ring.
Erythromycin, the first macrolide discwered, has been used since the early 1950s for
the treatment of upper respiratory tract and skin and soft tissue infectiors caused by
susceptible organisms, especially in the penicillin-allergic patient.
The soil-dwelling bacterium Saccharopdyspora eMhraea natively produces erythromycin A.
Additionally, erythromycin is effective for the treatment of infections caused by some
intracellular pathogens, induding species of Legionella, Mycoplasma, and Chlamydia.
Several drawbacks, however, have limited the use of erythromycin, including

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frequent gastrointestinal intokrance and a shortserum half-lift.

 Advanced macrolide antimicrobials have been synthesized by altering the emhromycin

base resulting in compounds➡️
 with extended spectrum of activity (clarithromycin and azithromvcin)

Macrolides action summarize by Binding to 50s subunit of ribosomes causing them to

dissociate from the mRNA resulting in premature termination of the amino acid chain &
cessation of protein synthesis

Other A.B.:

Lincomycin& Clindamycin: derived by combination of amino-acid and CHO metabolism.

Dactinomycin: anti —uneo plastic agent or A. B derived from a. a. metabolism (tryptophan)

Cycloserine: derived from a. a. metabolism

Vidarabine: derived from a. a. metabolism

Bictracin: derived from a. a. metabolism

Doxorubicin: derived from acetate metabolism (anti-cancer A.B.)

Mitomycin: anti — neo plastic A.B derived from acetate metabolism

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daunorubicin: anti — neo plastic A.B derived acetate metabolism

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