Antimicrobial agents and

vaccinations
Cecilia Chan
Antimicrobial control
Agents that kill microbes = ‐cidal (e.g. bacteriocidal, fungicidal, virucidal)
Agents that inhibit microbial growth = ‐static (e.g. bacteriostatic, fungistatic, virustatic)

agent added agent added

Number of bacteria

Number of bacteria
存活的

Bacteriostatic Bacteriocidal
抑菌 殺菌
Antimicrobial control

Antimicrobial agents
• natural or synthetic chemicals that kill or suppress growth of microorganisms

Antibiotics
• Originally refer to those naturally occurring microbial (mainly antibacterial) products
• With the presence of semi-synthetic antibiotics, the definition is now less well-defined
Antimicrobial agents for external use
Sterilants
• Kill all microbes including spores
• e.g. radiation, steam over 100oC

Disinfectants
• Kill microbes but not necessarily spores
• e.g. bleach to disinfect floors & table

Antiseptics
• Kill microbes or inhibit microbial growth, nontoxic to living tissues.
• e.g. 70% alcohol, hydrogen peroxide (雙氧水) & PVP-iodine (聚维酮碘)

Sanitizers
• Reduce microbial numbers to safe levels.
• e.g. sanitizers for hands, dishes, and utensils
“microbes are educated to resist penicillin”
—Alexander Fleming, 1945
Development of Antibiotic Resistance in S . aureus
Discover
streptomycin
1944
Synthetic
sulfonamides
1935

First
vancomycin-
resistant
Enterococcus
faecium
1988 2002
1959

• Due to the uptake of β-lactamase gene, 40% of the clinical isolates were penicillin-resistant by 1950, and went up
to 80% by 1960
• In 1959, methicillin was introduced to treat penicillin-resistant S. aureus infections
• Consequentially vancomycin served as a “last line of defense” against MRSA for the next 40 years
• Linezolid, daptomycin are used to treat vancomycin-resistant bacteria
https://web.mit.edu/Angles/Oran_Payne.htm
Percentage (%)
Anti-microbial drugs

➢ Anti-bacterial drugs
➢ Anti-viral drugs
➢ Anti-fungal drugs
Anti–bacterial drugs

• Naturally occurring antimicrobial drugs: antibiotics

• Semi-synthetic antibiotics
• Synthetic drugs
Naturally occurring drugs: antibiotics

• < 1% are useful clinically in humans because of high toxicity

and poor cellular uptake

Penicillin Tetracycline Actinomycin

Synthetic drugs
• Mainly growth factor analogs
• structurally similar to growth factors but non‐functional in
bacterial cells → inhibit bacterial growth
• 3 types: amino acid analog, nucleic acid analog, sulfa drugs Amino
acid
• Not commonly used nowadays because of drug resistance analog

Sulfa drug

nucleic
acid
analog
DNA
Folate
synthesis
Mode of action of anti-bacterial drugs
• Non‐toxic chemical compounds that can be used in vivo
• Classified according to mechanism of action
DNA gyrase
(DNA replication) Inhibit DNA repair
Cell Wall Synthesis DNA-directed RNA polymerase
(antitumor)
RNA elongation
Cycloserine Nalidixic acid Rifampin
Quinolones Actinomycin
Vancomycin
Bacitracin * Ciprofloxacin
Novobiocin
* Streptovaricins

*
Penicillins
β-
lactam * Cephalosporins
Monobactams
Protein synthesis
(50s inhibitors)
Carbapenems
* Erythromycin (macrolides)
Chloramphenicol
Clindamycin *
Folic acid metabolism Lincomycin
Trimethoprim THF
Linezolid
*
Sulfonamides
Protein synthesis
(30s inhibitors)
DHF
Tetracyclines
Spectinomycin*
Streptomycin
Gentamicin *
Cytoplasmic membrane Kanamycin Aminoglycosides
structure & function Amikacin
Polymyxins Lipid Nitrofurans
*
Daptomycin
biosynthesis
Protein synthesis
Platensimycin
PABA (tRNA)
Cytoplasmic Cell wall
Mupirocin
membrane Puromycin

Madigan et al., Brock Biology of Microorganisms (2003)

Site of action – cell wall

• A good target because only bacteria, but not human cells, possess cell wall
• Allow selective toxicity without harming the host
• Usually regarded as narrow-spectrum antibiotics, because only effective on a selected
group of bacteria
• Includes
o β-lactam agents: penicillin, cephalosporin, methicillin
o Glycopeptides: vancomycin
β–lactam antibiotics: Penicillins
β–lactam antibiotics
• Inhibitors of bacterial cell wall synthesis

• Characterized by the presence of β-lactam ring

• DD-transpeptidase is
needed for cross-linking the
polysaccharide chain
• Penicillins inhibit the (PBP)
(PBP)
reaction and block cross-
linking of the cell wall

DD-transpeptidase = penicillin-binding protein (PBP)

β–lactam antibiotics: Penicillins
β–lactam antibiotics
• Bacterial cell wall synthesis inhibitors

• Characterized by the presence of β-lactam ring

• Penicillin was isolated from fungus Penicillium chrysogenum

• Natural penicillin:
• Effective against Gram +ve and a few Gram ‐ve bacteria
• Render ineffective to bacteria expressing the resistance gene β-lactamase
(penicillinase)

• Semi‐synthetic penicillin:
• can be effective against some Gram ‐ve bacteria
• Many are β-lactamase-resistant
• Not bound and hydrolysed by penicillinase
• E.g. methicillin, oxacillin, ampicillin
β–lactam antibiotics: Penicillins
β–lactam antibiotics
• Bacterial cell wall synthesis inhibitors

• Characterized by the presence of β-lactam ring

• Resistance to methicillin is conferred by the macA gene

• macA gene encodes penicillin-binding protein 2a (PBP2a)

• PBP2a has low binding affinity towards β–lactam antibiotics

• Thus, β–lactam antibiotics is not working for macA-

expressing bacteria, e.g. MRSA
• MRSA cannot be effectively treated with antibiotics
such as methicillin, nafcillin, cephalosporin or
penicillin.
β–lactam antibiotics: Cephalosporin & Carbapenem
Cephalosporins
• Isolated from fungus Cephalosporium species
• β-lactam ring is fused to a six-membered ring
• Broader spectrum (Gram +ve and Gram –ve)
• Used to treat penicillin‐resistant infections (e.g. gonorrhea, E. coli)
• Cephalosporin is susceptible to β-lactamase (extended-spectrum beta-lactamases)

• Semi‐synthetic cephalosporins, e.g. ceftriaxone, is resistant to β-lactamase

• The new generation can treat MRSA

Carbapenems
• Broad spectrum, for treating both Gram +ve and Gram –ve bacteria
• Often reserved for serious infections caused by multidrug-resistant bacteria
 Glycopeptides: Vancomycin

• Last-resort antibiotics against serious infection, including MRSA

• Only active against gram-positive bacteria
• It is effective for streptococci, enterococci, and methicillin-susceptible
Staphylococcus aureus (MSSA) infections.

Daptomycin and linezolid could be

used to treat vancomycin-resistant
diseases.

Daptomycin – target bacterial cell membrane

Linezolid – target protein synthesis
Site of action – protein synthesis

* *

*
* *
Aminoglycosides
Target protein synthesis

• Produced by the bacteria, Streptomyces species

• Streptomycin, the first antibiotic in the group
• For treating enterobacteria and M. tuberculosis

• Not effective against Streptococci

• Binds to the 30S ribosomal subunit irreversibly → interfering the formation of
ribosomal complex → inhibit protein synthesis
Tetracyclines
Target protein synthesis

• Produced by the bacteria, Streptomyces species

• have a four‐ring structure
四環素
• Broad-spectrum
• For treating both Gram +ve and Gram –ve bacteria, mycoplasma, malaria and others
• Prevent tRNA binding to ribosome → inhibit protein synthesis
• Impair bone & teeth development in infants & children (<= aged 8)
• EXCEPT doxycycline if in short course, for which binds poorly to Ca2+
Site of action – nucleic acid synthesis

• Quinolones target DNA gyrase to inhibit DNA replication

• E.g. ciprofloxacin
• Broad spectrum
Site of action – cell membrane

Daptomycin
• Against Gram +ve bacteria
• Mainly used for vancomycin- and methicillin-resistant Staphylococcus aureus
Anti-microbial drugs

➢ Anti-bacterial drugs
➢ Anti-viral drugs
➢ Anti-fungal drugs
Anti-viral drugs

• Since virus “hijacked” the human cell for the production of viral particles, most antiviral
drugs also target human cells → potentially high toxicity
• Design of compounds that are selectively toxic for viruses (but not the host cells) is
crucial
Types of antiviral drugs
• Nucleoside analog
• Reverse transcription (RT) inhibitor
• Protease inhibitor
• Fusion inhibitor/ Entry inhibitor
• Adamantane
• Neuraminidase inhibitor
• Interferon
Nucleoside analogs &
Reverse transcription (RT) inhibitors
• Nucleoside analog
• The most successful class of anti‐viral drugs
• Its structure mimics the building block of DNA, i.e. A/T/C/G

• Reverse transcription (RT) inhibitors

• Reverse transcriptase does not exist in human cells
• Reverse transcriptase expression is only limited to viral infected cells
• The inhibitors bind to reverse transcriptase to inhibit its activity
Nucleoside analogs &
Reverse transcription (RT) inhibitors
• Azidothymidine (AZT)
• AZT is a thymidine analogue (nucleoside analog)
• works by selectively inhibiting HIV's reverse transcriptase (RT inhibitor)
• Prescribed for HIV

• Nucleoside analog – Acyclovir

• inhibits and inactivates the viral DNA polymerase
• Prescribed for herpes simplex virus, shingles

• Non‐nucleoside RT inhibitor – Nevirapine

• bind directly to reverse transcriptase
• Prescribed for HIV
Protease inhibitor

• inhibit viral maturation by preventing protease (e.g. HIV protease) to break down large
viral proteins into functional viral proteins
• Lopinavir
• Prescribed for HIV

Viruses 2023, 15(3), 712

Fusion inhibitor/ Entry inhibitor

• Block virus penetration into host cells (e.g. HIV penetration into T lymphocyte)
• Enfuvirtide
• Prescribed for HIV
• used in treating experienced patients that usually failed previous antiretroviral combinations
Influenza antiviral agent

• Adamantane
• synthetic amine
• intervene with viral uncoating & replication

• Neuraminidase inhibitor
• blocks active site of neuraminidase
• inhibit release of influenza virus
• e.g. Tamiflu, Zanamivir
Interferons (INFs)

• Cytokines release by the immune system in response to viral infection

• Interferes viral replication by inducing production of antiviral proteins from uninfected
cells
• Type I INFs are prescribed in supressing HCV and HBV infection, in combination with
other drugs

Cytokine Growth Factor Rev. 2014 Aug; 25(4): 369–376

List of common anti-viral drugs (I)

(Tamiflu)
List of common anti-viral drugs (II)
Anti-microbial drugs

➢ Anti-bacterial drugs
➢ Anti-viral drugs
➢ Anti-fungal drugs
Anti–fungal agents

• Fungi are eukaryotes. Therefore, it is challenging to develop antifungal drugs that

selectively target fungal cells without harming human cells.
• Most anti-fungal agents are used topically (i.e. on body surfaces)
• Effective anti-fungal drugs are those target structures or metabolic pathways specific
to fungi
• Classes:
• inhibit the synthesis of ergosterol, a vital component of the fungal cell membrane
• bind to ergosterol in the fungal cell membrane
• target the fungal cell wall by inhibiting the synthesis of beta-glucan, a key component of the
cell wall
• Others: interfere nucleic acid synthesis, interrupt microtubule formation
Anti-fungal drugs – mechanism of action

• Inhibit ergosterol synthesis

• Azoles
• Allylamines

• Bind to ergosterol
• Polyenes, e.g. amphotericin B

• Inhibit cell wall synthesis

• Echinocandins (target β-glucan)
• Polyoxin (target chitin)

Amphotericin B is often used to treat

severe or systemic fungal infections.
Drug effectiveness and resistance

Ineffective
• Lack of target structure of an antibiotic. For example, mycoplasma do not have
bacterial cell wall, so resistant to penicillin
• Impermeability to antibiotic (e.g. Gram –ve bacteria are impermeable to penicillin)

Resistant to drug
• enzyme inactivation (e.g. β‐lactamase secreted by Staphylococcus can cleave the
β‐lactam ring of penicillin)
• active efflux (pump out) to prevent cell entry (e.g. tetracycline efflux by E. coli,
Salmonella, Shigella)
Antimicrobial drug resistance

The resistance genes may

come from R plasmids,
transposons etc.

Lade, H. et al., Bacterial Targets of Antibiotics in Methicillin-Resistant Staphylococcus aureus. Antibiotics (2021)
Antimicrobial drug resistance
What kind of people are vulnerable to MDRO ?

People are vulnerable to multi-drug resistant organisms when :

1. there is an open wound (self-inflicted or by surgery)

2. there is an invasive device (intravenous catheter, urinary catheter) in place
3. they have compromised or weakened immunity
Emergence of antibiotic‐resistant pathogens

Due to improper or excessive use of antibiotics

Possible solutions:
1. only use drugs to treat susceptible diseases
2. administer drugs in sufficiently high / optimized doses & duration, so resistant strains
cannot develop
3. combine mechanistically unrelated antimicrobials agents
4. re-introduce drugs which were previously withdrawn
• Strengthen knowledge through surveillance and research;

• Optimise use of antimicrobials in humans and animals;

• Reduce incidence of infection through effective sanitation, hygiene and preventive measures;

• Improve awareness and understanding of AMR through effective communication, education and training;

• Promote research on AMR; and

• Strengthen partnerships and foster engagement of relevant stakeholders.

Topmost dispensed antimicrobial in non-
inpatient service in HK (2018)
A story of success

• In the southwestern U.S. and Mexico, brown dog ticks can carry a germ that
causes Rocky Mountain spotted fever in people and dogs.
• In Arizona, free-roaming dogs were spreading infected ticks. Many people got
sick and some died from Rocky Mountain spotted fever.
• Public health and animal health officials used long-lasting tick collars on dogs,
regular pesticide applications around homes, community education
• After only 4 months, 99% of dogs were tick-free in the community. People
infected decreased in number.

Doxycycline is used to treat

Rocky Mountain spotted fever
Vaccination
Vaccination

• Vaccination is an active immunization induces long-lasting immunity by stimulation of

antibody responses

An ideal vaccine
➢ Lifelong immunity
➢ Safety: no side effects
➢ Efficacy: check antibodies level
Herd immunity

• When most people in a community are immune to a particular infection → the natural
spread is limited
• Works only for infections transmitted from person to person
• E.g. does not work for tetanus
• Effective herd immunity: vaccine uptake rates >90%
• For highly transmissible infections: vaccine uptake rates >95%
Types of vaccine

• Killed or inactivated vaccines

• Live or attenuated (weakened) vaccines
• Toxoids vaccines
• Subunit / recombinant vaccines (purified
antigens)
• Viral vector vaccines
• mRNA vaccines Using a whole virus Protein that trigger A DNA/RNA that
encodes the
or bacteria the immune system
immunogenic protein
Killed or inactivated vaccines

• Use the killed version of the germ that causes a disease

• Microbes are treated with gamma radiation or a chemical (e.g. formaldehyde)
☺ Pros: much safer than live vaccines
 Cons: shorter durations than live vaccines; less effective so may need booster
• E.g. HepA, rabies, flu, polio
Live or attenuated vaccines

• For some microbes, inactivation may destroy the antigenic component, then live
vaccine is used
• Live microbes with reduced/weakened virulence
• Can still replicate in the recipient
☺ Pros: very effective
 Cons: risk of infection, should not be used on pregnant women or
immunocompromised individuals
• E.g. chickenpox vaccine (水痘疫苗), measles, smallpox
Toxoids vaccines

• Use a modified or inactivated toxin (toxoid)

• Toxoid preserve the antigenicity, but lost the toxicity
• For diseases that the bacterial toxin is causative agent
☺ Pros: more safe than live vaccine
 Cons: only effective if the disease is entirely attributed to toxin
• E.g. tetanus, diphtheria
Subunit / recombinant vaccines

• Antigen is expressed (in eggs / in cultured cells) and purified

• Antigens can be peptides or polysaccharides

☺ Pros: more safe than live vaccine ; can also be used on almost everyone, including
people with weakened immune systems and long-term health problems
 Cons: least effective ; need repeated doses

• E.g. Pneumococcal disease, HPV, Hepatitis B, Haemophilus influenzae B

Viral vector vaccines
• Developed based on molecular biology advances
• Use a modified, harmless virus (e.g. adenovirus in common cold) as a vector
• Replace the disease-causing genes in a virus with the gene encoding the antigen
• The viral vector infect human cells and instruct them to make large amounts of antigen
• Two main types of viral vector-based vaccines: non-replicating and replicating
• E.g. COVID-19 (Johnson & Johnson ; Oxford/AstraZeneca), Ebola
☺ Pros:
• Easy to produce
• Can trigger strong immune response
• No need to freeze, only refrigerate
 Cons:
• If a viral vector vaccine uses a virus that one has already been exposed to, the vaccine won’t be as
effective
• Side effects
mRNA vaccines

• mRNA strands are injected instead of pathogens (treated) or proteins

• mRNA makes proteins, which is responsible for triggering an immune response
☺ Pros:
• shorter manufacturing times
• because they do not contain a live virus, so no risk of causing disease
 Cons:
• Genetic sequence of the pathogen must be known
• Must be stored in freezer
• E.g. COVID-19 (Pfizer-BioNTech ; Moderna)
• This technology was under research until COVID-19 that was brought to application for the
first time
Booster shots

• Vaccines works to trigger the adaptive immunity to produce antibodies

• Over time, antibody levels can naturally decrease, and the immune response can fade
• Booster shots are required to restored the protection
• Antibody levels can be measured to check if booster shot is needed

• The reason why acquired immunity lasts longer for some vaccines than others is not
fully understood
• For pathogens that mutate frequently
• E.g. Flu, COVID-19
 Booster shots
• Influenza virus • SARS-CoV-2
• Mutations of HA and NA • Mutations of spike protein

Mutations caused by antigenic drift and antigenic shift

https://www.drugoffice.gov.hk/eps/do/en/consumer/news_informations/dm_38.html
THANK YOU!
Questions welcome