Erythromycin

Christine Lachnit
Doreen Könning
Marie Liebig
Patrick Klink

Fachbereich Chemie
TU Darmstadt
1 Introduction
Erythromycin is an antibiotic which belongs to the
group of macrolide antibiotics. The pharmaceu-
tically distributed product consists of three com-
ponents: Erythromycin A, B, and C where Ery-
thromycin A represents the main component. Nat-
urally this antibiotic is synthesized by the gram-
positive bacteria Streptomyces erythreus (Saccha-
ropolyspora erythrea).

2 History
In 1949 Erythromycin was found for the first time
Figure 1: Structure of Erythromycin
in a soil sample in the Philippine region Iloilo. A
research team, led by J. M. McGuire, was able to
Type R1 R2
isolate Erythromycin which was part of the soil
A -OH -C H3
sample.
B -H -C H3
Under the brand name Ilosone the product was
C -OH -H
launched commercially in 1952. They named the
brand after the region where the antibiotic was Table 1: Different types of Erythromycin
found. Analogically the first product name was Ilo-
tycin. Furthermore, in 1953 the U.S. patent was
granted. 4 Chemical and Pharmacokinetic Properties
Since 1957 the structure of Erythromycin is
known and in 1965 the X-ray structure analysis Formula: C37 H67 N O13
gave awareness of the absolute configuration. CAS-Number: 114-07-8
In 1981, almost 30 years after the detection Molar Mass: 733.93g/mol
of Erythromycin, Robert B. Woodward, the Nobel Half Hife 1.5 hours
prize laureate of chemistry in 1965, and his co- pkA: 8,6 - 8,8
workers posthumously reported the first synthesis Melting Point: 411K (hydrat)
of Erythromycin A. 463-466K (anhydrous)

3 Structure Table 2: Chemical and Pharmacokinetic Properties

The structural characteristic of macrolides, to
which Erythromycin affiliates, is a macrocyclic lac-
tone ring of fourteen, fifteen or sixteen mem- 5 Biological Effect
bers. In case of Erythromycin the lactone ring Antibiotics are substances which inhibit the growth
consists of 14-members. Substituents on the main- of microorganisms or even kill them and thus are
chain are cladinose on C-3 and desosamine on C-5. often used in order to combat bacterial infections.
Erythromycin is not a single compound but rep- There are different points of attack, for example:
resents an alloy of structural very similar compo-
nents. The main constituents are Erythromycin A, • cellwall
B and C. As shown in Table 1 and Figure 1 they
• biosynthesis of cellwall
only differ in two rests on the lactone ring or on
the cladinose each case. • transcription
In addition to the variants already mentioned,
further variants, like Erythromycin D and E are • protein biosynthesis
known. They are pre- and post-stages in the • DNA replication
biosynthesis and often do not have antibiotic ef-
fects. • metabolism

1
Erythromycin, as well as all the other macrolides 7 Derivatives
and ketolides (in Section 7), inhibit the protein
biosynthesis - especially of gram-positive bacteria Erythromycin is degradable in acidic environments
- in the ribosomes of the bacterial cells. They due to its structure which is disadvantageous as it
reversibly bind the 50S subunit of the 70S ribo- gets decomposed in the stomach if administered
somes such that the complex consisting of tRNA orally. Furthermore, the antibiotic spectrum does
and aminoacids is fixed at the A binding site. The not involve many gram-negative bacteria, which
outcome is prevention of the transfer to the E often cause diseases. In addition to those debilities,
binding site and thus the following reading of the Erythromycin is less effective than other antibiotics
mRNA. Finally the protein biosynthesis stops pre- like Penicillin. In order to improve the proper-
maturely. ties of Erythromycin, a chemical modification of
its structure was necessary and desirable. Until
6 Resistance the 1990s scientists were not able to modify Ery-
thromycin specifically. The received semisynthetic
Bacteria are able to develop various resistances derivatives contained better pharmacological prop-
against antibiotics, for example, by mutation or erties and, thus, are preferred in the medical appli-
horizontal gene-transfer. The following resistance ance. The advantages over Erythromycin are an
mechanisms are known, possibly in combination: improved stability in acidic environments, avoid-
ing resistances, and a longer elimination half-life
• modification of the target site
in human metabolism. As a result of the deriva-
• enzymatic inactivation of the antibiotic tives’ characteristics, the antimicrobial spectrum
has increased and the minimal inhibitory concen-
• active elimination of the antibiotic tration has been lowered. Hence reduced doses of
antibiotic and less days of consumption are pos-
• inhibited absorption of the antibiotic
sible. Important derivatives of Erythromycin are
Against macrolide antibiotics there are two mech- other macrolide antibiotics and ketolides.
anisms of resistance: active efflux (particularly
streptococcus) or target site modification. They 7.1 Ketolide
are known as M-resistance respectively MLSB- The ketolid antibiotics are derivatives of Ery-
resistance. thromycin A. The structural difference is an ex-
change of the cladinosyle group with a keto goup
M-Resistance at the C3-Atom. This keto group is responsible for
This type of resistance is the more common the diverse mechanism of action and the missing
but less effective mechanism by building an efflux MLSB-resistance of ketolides. Thus, the reason for
pump. The coding gene is the Macrolide-Efflux- the macrolides’ induction of resistance supposably
Gene (mef(A)). In order to get subinhibitory con- is the cladinosyl group. Furthermore, the keto
centration of antibiotics with this pump, bacteria group causes a better stability in acidic environ-
reject the macrolides out of the cells. ments and avoids the induction of resistances men-
tioned above. For the synthesis of ketolides, a pre-
MLSB-Resistance stage of Erythromycin A (a 6-O-methylanalogues)
The letters M,L, S and B stand for Macrolides, is used. In an acidic milieu the cladinosyl group is
Lincosamides and Streptogramin type B. This more removed. Via a 3-hydroxy-intermediate the keto-
effective mechanism of resistance differentiates the group is generated. This skeletal structure can
ribosomes’ receptors for (in this case) macrolides. further be modified, for instance, with a carba-
An adenine of the 23S-RNA gets dimethylated by mate group at C-11 and C-12 as it is the case
methylases such that macrolides cannot bind any with Telithromycin. It holds a better antibiotic ac-
longer. The responsible gene is the Erythromycin- tivity against Erythromycin A-resistant cocci. In
Methylase-Gene (erm(B)). difference to other ketolides, Telithromycin has a
methoxy group on C-6 instead of a hydroxyl group.
This inhibits a ketalization with the keto group on

2
C-3. The ketolides mechanism of action is similar completely be resorbed from the gastrointestinal
to the macrolides one but it differs in the interac- tract. Moreover, it diffuses into most tissues and
tions with the bacterial ribosomes. Based on the it features a longer half-life period. Roxithromycin
keto group on C-3 or the carbamate group on C- can also be used against gram negative bacteria
11 and C-12 (Telithromycin) the molecule binds on such as legionellas and neisserias.
the target site ten-times stronger than macrolides.
Moreover ketolides have a direct impact on the for- Azithromycin
mation of the 30S and 50S ribosomal subunits. In Azithromycin also is a semisynthetic modifica-
other words ketolides use a dual mechanism of ac- tion of the macrolid antibiotic Erythromycin. They
tion. differ structurally in the enlargement of the lacton
ring at the C-9 atom. Azithromycin posseses an
7.2 Macrolide N-methylaminomethylen-group, which causes a 15
membered instead of a 14 membered ring. The ni-
Clarithromycin
trogen atom is responsible for the higher basicity of
Clarithromycin, a semisynthetic macrolide, has
the molecule. To emphasize these differences com-
been developed in the 1970s and is gained from the
pared with Erythromycin, Azithromycin is called
naturally occurring macrolide Erythromycin. Ery-
an azalid antibiotic. Until today, it is the only
thromycin and Clarithromycin differ structurally in
representative of this category of antibiotics. As
a methoxy-group on C-6 (Clarithromycin) instead
all derivatives, Azithromycin reveals a higher acid-
of a hydroxyl-group (Erythromycin). The struc-
stability and the antibacterial effect on bacteria is
tural difference leads to a higher acid-stability of
quite the same as with most macrolids. In con-
Clarithromycin. Due to its acid-stability, it is re-
trast to Erythromycin, it has a considerable effect
sorbed better from the gastrointestinal tract with-
on gram negative bacteria such as Haemophilus in-
out having to be refurbished previously. Therefore,
fluenza and because of its long half-life period it is
it can be taken orally without being protected from
used as three-day antibiotic. Furthermore, it also
the gastric acids. A further advantage over Ery-
differs quickly into most tissues.
thromycin is the longer half-life period which leads
to a possible appliance of lower daily-doses. More-
over, the minimal inhibitory concentration of Clar- 8 Medical Relevance
ithromycin is lower for many gram-positive bacte- Erythromycin and its derivatives are mainly used
ria compared to Erythromycin and its antibacterial for the treatment of diseases caused by gram-
effect on gram-negative bacteria is better. In ad- positive bacteria such as Staphylococcus or Strep-
dition, it affords an improved compatibility. How- tococcus. Some gram-negative bacteria, for in-
ever Erythromycin-resistant streptococci are not af- stance, Chlamydia and Legionella can be medi-
fected by this antibiotic. cated as well.

Roxithromycin Excursion: Gram-positive and -negative Pathogenes

Roxithromycin aswell as Clarithromycin is a Bacteria are divided in two large groups, based
semisynthetic macrolid antibiotic that was first on their behaviour in gram-staining. The dif-
developed in 1991. The structural difference ference lies in the chemical and physical
compared with Erythromycin, is the functional properties of the cell walls. The cell walls of
group at the C-9-atom: an oxim group instead all bacteria consist of a cytoplasma membrane
of a keto group is revealed (Ery-9-[(E)-O-([2- (inner membrane), that seperates the cyto-
methoxyethoxy]methyl)oxim]). The mechanism of plasm inside the cell from the environment.
action is the same as with all macrolides. It inhibits But gram positve and gram negative bacteria
the protein biosynthesis. Besides Roxithromycin clearly differ in the remaining structure of
exhibits a similar antimicrobial spectrum. Gram their cell walls. Gram-positive bacteria own a
positive bacteria are affected by lower concentra- multilayered murein sacculus (cell wall con-
tions of this macrolid antibiotic as it is the case sisting of peptidoglycan) that is responsible
with clarithromycin. Compared with Erythromycin for the cell’s shape but permeable for antibi-
the acid-stability is improved and, therefore, it can otics. Besides a thin murein sacculus (max. 2

3
 layers) gram negative bacteria on the other and Azithromycin. Resistance problems are yet
hand feature an external membrane, that is very rare. In contrast, marolids are less effective
a diffusion barrier to antibiotics. That is the when very dangerous and unusual eye socket in-
reason why gram-positive bacteria are treated fections occur. These can be caused by sinusitits
easier with antibiotics. By means of the gram and, therefore, the macrolid antibiotics have to be
staining, the difference in the structure of the replaced by more efficient antibiotics. As an alter-
cell wall can be made visible. The method is native to Penicillin for instance, macrolids can be
based on the fact that a stain applied on gram used for Otilis Media, Pharyngitis, and Tonsillitis.
positive bacteria and permeated inside the cell However, it is possible that modifications of those
cannot be removed because of the thickness diseases should not be treated with macrolid an-
of the cell wall. This is opposite to the case tibiotics at all.
with gram negative bacteria.
8.2 Lower Respiratory Tract Infections
Erythromycin is of great importance due to the
Pneumonia: Macrolids in particular are very advan-
good compatibility. Only few side effects are
tageous in the treatment of "Community Acquired
known, such as light indigestions. This is caused
Pneumonia" (CAP), a form of pneumonia, that is
by the abilitiy of Erythromycin to act as a motilin-
acquired outside of hospitals. However, the domi-
receptor-agonist. Motilin is a gastrointestinal pep-
nant bacteria have to be inclosed in the macrolid’s
tid hormon, that stimulates the gut motility. The
antimicrobial spectrum. Bacteria based pneumo-
antimicrobial spectrum of Erythromycin is similar
nias can be caused by many different bacteria. In-
to those of some penicillins. Penicillins belong to
dications of Streptococcus pneumoniae, which are
the category of β -lactam antibiotics. Therefore,
sensitive to β -lactam antibiotics, are soft crackles
Erythromycin is often used for people who have
for instance. Since it is very hard to diagnose ex-
an allergy to this type of antibiotics. Another ap-
actly which bacteria are involved, there are many
pliance is the use against penicillin resistant patho-
strategies for the use of antibiotics. One possibilty
genes.
is to start with β -lactam antibiotics and switch to
In general, the effectiveness of Erythromycin is
macrolids, if no clinical success is revealed. More-
slightly decreased compared to Penicillin. Further-
over, a combination of several different antibiotics
more, Erythromycin is a weak alkali and, thus,
is imaginable. Especially babys and toddlers are
forms salts and esters in organic acids. So called
treated with macrolids when pneumonia is diag-
Erythromycin alkalis can be established in the
nosted. Particularly an infection caused by chlamy-
blood. The gastric acid inactivates these alkalis,
dia at the childbirth comes within the limits of
esters, and salts differently.
macrolids. Infection of pseudonoma with cystic fi-
Erythromycin is used for respiratory tract infec-
broses: Although macrolids are not very efficient in
tions, infections of the skin as well as otolaryngo-
the therapy of pseudonomas, addition of macrolids
logical infections.
such as azithromycin showed a positive effect on
patients with cystic fibroses. Combined with the
8.1 Upper Respiratory Tract Infections
standard therapy, which includes the intake of a
Sinusitis (inflammation of the paranasal sinuses): A combination of an amino glycosid with a β -lactam
viral infection is the condition for an acute sinusi- antibiotic, those microorganisms are reached much
tis. As a consequence mainly unspecific defence better. Marolids inhibit the bacteria’s virulence fac-
mechanisms, for instance the viscosity of the mu- tors and additionally feature better immunomod-
cosa slime layer is disturbed. Infected epithel cells ulatory properties. Treatment of pertussis and
are settled at a concentration 100 times higher. bronchitis: After the medication with other antibi-
Combined with the cytokine induced swelling of otics was unsuccessful, babys can be treated with
the mucosa and the resulting stop of secretion a macrolid antibiotics when pertussis or bronchitis
super infection of the paranasal sinuses may occur occur.
with babys and toddlers. The responsible bacte-
ria can be treated with Erythromycin, but essen-
tially Clarithromycin, Josamycin, Roxithromycin

4
 8.3 Infection of the Skin and Soft Tissues in form of carbon dioxide is eliminated. The first
stable and enzyme free intermediate after the cir-
Impetigo: Impetico contagiosa is a skin infection cularization is 6-Deoxyerythronolid B as shown in
caused by β hemolysed streptococci which leads Figure 2.
to a muliplication of the resident germs at the in-
fected area. After an isolation of the pathogens and
a resistance test, marcolids, especially Josamycin
are administered.

Acne: Propionibacterium acnes leads to a de-

composition of the sebaceous gland’s fats in
glycerin and fatty acids, that cause an inflamma-
tion of the sebaceous glands. Staphyolocooci and
other bacteria can spread over the aggrieved skin
unhinderedly. Macrolids have anti-inflammatory
properties and accumulate on the skin 2-5 times
better than in blood. Therefore, they are predesti-
nated for the application of acne and many other
skin infections. Figure 2: Structure of 6-Deoxyerythronolid B

9 Synthesis
Phase 2
Erythromycin can be manufactured in two different Starting with 6-Deoxyerythronolid B, the ac-
ways. Naturally the antibiotic is produced by the tive antibiotic is produced with the aid of sev-
bacterium Streptomyces erythreae. This procedure eral different enzymes. First, a hydroxylation
describes the biosynthesis of Erythromycin. For catalyzed by the C-6 erythronolide hydroxylases
decades chemistrists dealt with the chemical syn- (eryF) at the C-6 occurs. The absolute configu-
thesis of Erythromycin, which only provided very ration of the stereogen center does not change in
small yields. Currently, the fermentative produc- this step. Next, a glycosylation with L-Mycarose
tion via bacteria is the best possibilty to produce at the C-3 via thiamindiphosphat-mycarose gly-
Erythromycin comercially. cosyltransferases follows. In the following step,
a D-desosamine is attached to the C-5 atom
9.1 Biosynthesis via glycosylation as well. The involved en-
The exact mechanism of the biosynthesis of Ery- zyme is thiamindiphosphat-desosamine glycosyl-
thromycin is not clarified yet, especially concern- transferases. The obtained product is Erythromycin
ing the sugar residues. The whole synthesis can D, which is the first intermediate of the biosynthe-
be divided in two phases. During the first phase sis that holds an antibiotic effect. For the further
the circularization, which is catalysed by one sin- synthesis of Erythromycin A, two paths are possible
gle enzym, takes place. In the second phase, six at which one is highly preferred over the other. The
enzymes are needed for the completion the antibi- preferred way begins with a hydroxylation at the C-
otic molecule. 12, which leads to the intermediate Erythromycin
C. The absolute configuration at the carbon atom
Phase 1 is retained during this process. Afterwards an O-
The participating enzyme in Phase 1 is the methylation via O-methyltransferases and SAM (S-
polyketid synthethases, which catalyzes the syn- adenosyl methionine) at the hydroxyl group of the
thesis of the lacton ring. The mechanism of reac- mycarose residue on the C-3 atom takes place. The
tion is similar to the synthesis of fatty acids via the less favored path runs conversely with the same en-
enzyme fatty acid synthases. First of all 6 units zymes and reactions involved. The final product is
of 2-methylmalonyl-CoA (activated thioester) are Erythromycin B.
added to one molecule propionyl CoA. During the
subsiding condensation reaction, a carbon atom

5
 9.2 Chemical Synthesis 5. J. Mulzer, Die Erythromycin-Synthese - eine
unendliche Geschichte?, Angewandte Chemie,
The chemical synthesis of Erythromycin poses a Volume 103, Wiley-VCH, 1991
huge challenge. The molecule contains ten stere-
ogenic centers of which five are arranged consecu- 6. J. Guggenbichler, Makrolid-Antibiotika,
tively. R. B. Woodward and his research team first accessed online June 2010,
succeeded in synthesizing Erythromycin A. The re- http://www.antibiotikamonitor.at
action sequence, however, is so complicated that
the yield was only about 0,02 % and, thus, the 7. S. Pal, A journey across the sequential devel-
synthesis is not utilizable comercially. This is the opment of macrolides and ketolides related to
reason for the preferred use of the biosynthesis of erythromycin, Tetrahedron, Volume 62, Num-
Erythromycin via fermentation of Streptomyces ery- ber 14 , pages 3171–3200, Elsevier, 2006
threus. Other scientists and research teams dealt 8. B. Weisblum, Erythromycin resistance by ribo-
with the synthesis of Erythromycin as well and de- some modification, Antimicrobial Agents and
veloped very similar approaches. Most methods for Chemotherapy, Volume 39, Number 3 , pages
the Erythromycin synthesis are based on the con- 577, American Society for Microbiology, 1995
struction of the aglycon from secoic acid via gly-
cosylation. Indeed the process is also possible in-
versely: first, a glycosylation, then a lactonization
occurs. The yield, however, is considerably less.
While earlier scientist mainly dealt with the pro-
duction of the different secoic acids, the lactoniza-
tion process is the major problem today because
there is no fully developed method for it yet. A
lot of side reactions such as dimerization and poly-
merization appear, because a 14 membered ring is
hard to enclose. Even if the chemical synthesis of
Erythromycin has no importance for the comercial
fabrication of the antibiotic, it is still important for
the development and fabrication of its derivatives.

10 References
1. B. Rawlings, Type I polyketide biosynthesis in
bacteria (Part A—erythromycin biosynthesis),
First published as an Advance Article on the
web March 2001

2. J. Staunton, B. Wilkinson, Biosynthesis of Ery-

thromycin and Rapamycin, Chem. Rev, Vol-
ume 97, Number 7 , pages 2611–2630, Amer-
ican Chemical Society, 1997

3. C. Khosla, Structures and Mechanisms of

Polyketide Synthases, JOCPerspective, Volume
74, Number 17 , pages 6416–6420, American
Chemical Society, 2009

4. D. Evans, A. Kim, Synthesis of 6-

Deoxyerythronolide B. Implementation of
a General Strategy for the Synthesis of
Macrolide Antibiotics, Tetrahedron Lett, 1997

6
A Appendix

Figure 3: Biosynthesis of Erythromycin