3. INTRODUCTION
• Macrolides:
• The macrolides are a class
of antibiotics that consist of a
large macrocyclic lactone ring to
which one or more deoxy sugar
attached.
• It includes erythromycin,
roxithromycin, azithromycin and
clarithromycin.
Erythromycin
4. MACROCYCLIC LACTONE
• Macrocycles are often
described as molecules containing
twelve or more membered ring.
• Lactone are cyclic carboxylic
esters.
• Deoxy sugars are sugars that
have had a hydroxyl group
replaced with a hydrogen atom.
DEOXY SUGAR
INTRODUCTION
5. MECHANISM
OF ACTION
• Macrolides work by
binding to a specific
subunit of ribosomes
(sites of protein
synthesis) in
susceptible bacteria,
thereby inhibiting the
formation of bacterial
proteins.
• The macrolides bind to
the 50S ribosomal
subunit.
6. INTRODUCTION
• Streptomyces caniferus:
• It is a bacterium species from the genus of Streptomyces.
• It is gram-positive bacteria.
• It is a filamentous bacteria.
• It is an aerobic bacteria.
• It has high CG content in their genome.
7. PM100117
• It is 36-membered macrolide with
a side chain containing 3-deoxy
ribose sugars and a 1,4-
naphthoquinone chromophore.
• It is reddish in color.
• Molecular formula is C82H130O29.
• It is 36-membered macrolide with
a side chain containing 3-deoxy
ribose sugars and a 1,4-
naphthoquinone chromophore.
• It is yellowish in color.
• Molecular formula is C82H130O28.
PM100118
DISCOVERED MACROLIDES
8.
9. METHODOLOGY
Isolation of Bacterial strain
• Streptomyces caniferus was isolated by spreading the homogenized marine polychaete, on plates of
Bennett′s agar medium.
• Medium is supplemented with nalidixic acid (0.02%) and cycloheximide (0.02%).
• The plates were incubated at 28 °C for 30 days.
• Phylogenetic analysis of the strain based on 16S rRNA gene sequence comparisons with databases.
Fermentation processes
• For the production of macrolides,12.5 ml of inoculum was transferred into flasks containing 250ml of
fermentation medium .
10. METHODOLOGY
• Fermentation medium containing yeast extract (0.5%), soya peptone (0.1%), dextrose (0.5%), soya flour
(0.3%), Glucidex (Roquette, France) (2%), sodium chloride (0.53%), potassium chloride (0.02%),
magnesium chloride 6.H2O (0.24%), sodium sulfate (0.75%), manganese sulfate 4.H2O (0.00076%),
cobalt chloride 6.H2O (0.0001%), di-potassium phosphate (0.05%) and calcium carbonate (0.4%).
• The culture was grown at 28 °C using an orbital shaker at 220 rpm for 5 days.
Extraction
• The fermentation broth subjected to centrifugation.
• The mycelial cake extracted by using isopropyl alcohol and ethyl acetate.
• The solvent is subjected to semi-preparative chromatography.
11. METHODOLOGY
• The fractions that eluting yields PM100117 (P1) and PM100118 (P2).
Assay of anti-proliferative activity
• Lung carcinoma, colorectal carcinoma, breast adenocarcinoma cell lines were obtained.
• Cell lines were maintained in RPMI medium supplemented with 10% fetal calf serum (FCS), 2mM L-
glutamine and 100 U ml− 1 penicillin and streptomycin, at 37 °C and 5% CO2.
• Triplicate cultures were incubated for 72 h.
• The cells were washed twice with PBS.
• Cells were then rinsed several times with 1% acetic acid solution and air-dried.
12. METHODOLOGY
• Three reference parameters were calculated by automatic interpolation:
• GI50= compound concentration that produces 50% cell growth inhibition, as compared with control
cultures;
• TGI= total cell growth inhibition (cytostatic effect), as compared with control cultures.
• LC50= compound concentration that produces 50% net cell killing (cytotoxic effect).
Assay of primary mechanism of action
• Lung cancer and breast cancer cells were exposed for 24 h to a 10 μM concentration of 1 and 2,
respectively. Representative phase contrast images of the experiment with both 1 and 2 showed
necrotic death with the features of plasma membrane permeabilization, including the formation of
13. RESULTS
• Both PM100117 (1) and PM100118 (2) have shown
potent antitumor activity against human lung
carcinoma cells, human breast adenocarcinoma cells
and human colorectal carcinoma.
• Permeabilize the plasma membrane.
• Also have shown slight antifungal activity against
Candida albicans.
• Researcher declares no conflict of interest.
14. REFERENCES
• Marta Pérez, Carmen Schleissner, Rogelio Fernández, Pilar
Rodríguez, Fernando Reyes1, Paz Zuñiga, Fernando de la Calle
and Carmen Cuevas, The Journal of Antibiotics, December,
2015.
• Omura, S. Macrolide Antibiotics: Chemistry, Biology, and
Practice 2nd Ed. (Academic Press, Boston, MA, USA, 2002)
• Papazisis, K. T., Geromichalos, G. D., Dimitriadis, K. A. & Kortsaris,
A. H. Optimization of the sulforhodamine B colorimetric assay. J.
Immunol. Methods 208, 151–158 (1997).
• Kim, O. S. et al. Introducing EzTaxon-e: a prokaryotic 16S rRNA
Gene sequence
• database with phylotypes that represent uncultured species. Int.
