ANTIBIOTIC PRODUCTION

INTRODUCTION

• Chemical subst. that inhibit growth and even destroy microorganisms

• Are the secondary metabolites of microorganisms
• Discovered using a screening process or a rational design process.
• Delivered into body thro. Absorption into bloodstream
• Destroy bacteria by affecting their structure
 HOW ANTIBIOTICS DESTROY BACTERIA

• Weakens the cell wall of bacteria causing them to burst

• Damage cell membrane, causing content of of bacteria to leak out
• Some interfere with bacteria metabolism
• Some interfere with protein synthesis e.g. tetracycline
• Some inhibit nucleic acid synthesis e.g. rifampin
 IDENTIFYING USEFUL BACTERIA
• Despite wide variety of known antibiotics, less than 1% antimicrobial agents have medical
or commercial value
• Some affect human cells, others lack advantage over those already in use, hence
eliminated
• Useful antibiotics discovered by screening
• Isolates of different microorganisms are cultured, tested for production of diffusible
products inhibiting growth of test organisms
• Remainders are tested for their selective toxicities and therapeutic activities
• Best candidate examined, modified. Grown in large scale, compound responsible for
antibiotic effect isolated
 RATIONAL DESIGN

• Modern approach for antibiotic identification

• Entails screening aimed at finding new natural products that inhibit a specific target e.g.
enzyme only found in target pathogen ,instead of testing a general inhibition
• Its more accurate since there are low chances of cross contamination, clear zones of
inhibition, efficient for minimum inhibitory concentration
• Scientist may use this technique to make desired changes based on knowledge of
structure and function of protein
• Drawback-Detailed structural knowledge of protein is unavailable,making it hard to predict
 FURTHER STUDIES

• In vivo studies done, using animal models like mice

• Pre clinical studies done using apes
• Clinical studies done using humans
• N/B: Ethical Clearance must be obtained prior to the beginning of in vivo studies
• Safety studies are also conducted: impact of the antibiotic on the human body, its
side effects, body reaction to the drug etc.
• If antibiotic is promising, the FDA approves antibiotic as new product
 INDUSTRIAL PRODUCTION

• 3 Techniques:
 Fermentation
 Semi-synthetic
 Synthetic
 SYNTHETIC TECHNIQUE

• Not all antibiotics produced by bacteria

• Antibiotics made in the lab, chemically synthesized based on the structure of the natural
products., antimicrobials
• Example is quinolones class, nalidixic acid, sulphonamides, cotrimoxazole
• Nalidixic acid was discovered by accident in lab during synthesis of chloroquine
• Advantages include: a broad spectrum of activity, rapid bactericidal activity, no cross-
resistance with the existing antibiotics and a low probability for developing resistance.
 SEMI-SYNTHETIC ANTIBIOTICS

• Combination of of natural fermentation and lab work to maximize the antibiotic

• Maximization occurs through efficacy of drug ,amt of abtc produced, potency (effectiveness
measure) of abtc being produced
• Example is Ampicillin. Was developed by addition of amino grp to the R group of penicillin.
• The addition gave ampicillin broader spectrum than penicillin
• Methicillin another example; is a derivative of penicillin. Addition of two methoxy groups to the
phenyl grp.
• Methoxy groups allow methicillin to be used against penicillinase producing bacteria which is
resistant to penicillin
 FERMENTATION ANTIBIOTICS

• For large scale production, scale up

• Source microbe grown in large containers/tanks (10000-150000) litres ,in presence of
liquid medium.
• Growth condition of microbes maintained/controlled; oxygen concentration, temperature,
pH, nutrients
• Population size of secondary metabolites must be controlled before cells die
• For new antibiotic to be feasible, manufacturers must get high yields of drug from
fermentation to easily isolate it
 FERMENTATION ANTIBIOTICS
• Large quantities of desired antibiotic excreted
• Tanks cooled to keep the temperature btn 23 to 27.2 degrees Celsius
• Constant agitation and continuous stream of sterilized air pumped in for homogenous
environment, sterility
• Antifoaming agents periodically added, after agitation
• Acids, bases added to control Ph (Must not interfere with tank contents,)
• Examples include: cephalosporins, chloramphenicol, Nisin, Erythromycin,Bacitracins
 RAW MATERIALS USED

• Compnds making the broth

• Broth contains all ingredients for microorganisms proliferation
• Carbon sources e.g. molasses, soya meal
• Lactose and glucose sugars
• Nitrogen sources ammonia salt used
• Trace elements-fastens growth of microbes
• Microelements e.g. phosphorus, sulfur, zinc-introduce through water soluble salts
• Antifoaming agents e.g. lard oil, octadecanoyls
 DOWNSTREAM PROCESS

• Isolation and purification of fermentation broth

• Compounds separated from waste organic material
• Water soluble compounds then obtained
• Water soluble antibiotics may be obtained thro. ion exchange technique,
• oil soluble antibiotic solvent extraction method used-broth treated with organic solvents
e.g. butyl acetate that dissolves antibiotics. Dissolved antibiotic recovered by various
organic chemical means
• Chromatography(HPLC) MAY BE USED,
 DOWNSTREAM PROCESS
• Desired products must be obtained using various downstream processes ion exchange,
adsorption, chemical precipitation, centrifugation
• Antibiotic must be extracted and purified to a crystalline product
• If antibiotics soluble in organic solvents it’s a bit easy to separate
• Extensive research conducted later for efficiency, potency
• Antibiotic modified into various biopharmaceutical formulations: tablets, capsules, injectable
formulations, liquid form
• Pharmaceutical excipients added e.g. fillers-add volume/mass o drug, binders-hold ingredients
of drug, mechanical strength of drug, coating agents –ease of
swallowing,protection,identification,preservatives-prolong shelf life, sweeteners, coloring
agents, buffering agents
 PACKAGING AND STORAGE

• Packaging done in containers, labelling done, manufacturer instructions attached

• Storage at specific temperatures, maintaining efficiency of the drug
• Transport
DIAGRAM EXPLANATION