PHAR1101 Week7 L1 – Penicillin (由青黴菌中提煉出的抗生素, 可以干擾細菌的細胞壁的合成- 有殺菌作用, 用於治療尿道炎、中耳炎、鼻竇

炎、支氣管炎、口腔感染和肺炎) & the Golden Age of Antibiotics

Aim of the lecture:
 Describe the development, significance, and
limitations of Salvarsan and sulfonamides 磺胺類
藥物(人工合成的抗菌藥物, 一類用於預防和治療
細菌感染性疾病的化學治療藥物) as antibiotics
prior to penicillin
 Describe key events and the roles of Fleming and
Florey in the discovery of penicillin
 List important scientists and their contribution to
the understanding of penicillin and the
development of improved penicillin derivatives
 Explain how penicillin works to kill sensitive
bacteria
 List three common mechanisms of drug resistance
 Describe the serious problems raised by the
emergence of drug-resistant microbes and the
challenges of addressing these problems
The Beginning of the antibiotics era
Two innovative synthetic drugs 合成藥物 found via animal
testing of “compound libraries”:
 1911 - Salvarsan 灑爾佛散 (Paul Ehrlich)
o Arsenic 砷 drug for treating syphilis
infectious 梅毒 (透過人類性行為傳播。
該疾病也可由母親在懷孕或分娩時傳染
給胎兒)
 1935 - Prontosil 百浪多息 (Gerhard Domagk)  Toxicity to patient with salvarsan (arsenic)
o Azo drug for streptococcal infectious A 型  Skin stained with Prontosil (not other
鏈球菌感染 sulfonamides)
How Sulfonamides 磺胺類藥物 (人工合成的抗菌藥物)  Bacterial resistance a problem with Sulfa drugs
paved 鋪好的 the way? The Penicillium Mould (A Goldmine for Drugs?)
 Proved that fatal infectious disease were  Antiquity 古代: Use of mouldy bread to treat
“manageable” with drugs e.g. pneumonia 肺炎 infected wounds (Greece, China, Egypt, etc)
 Fostered 培養 hospital microbiology infrastructure: o Anglo-Saxon recipe (1,000 yrs) recently
o Well equipped labs shown to kill MRSA (methicillin-resistant
o Handling lots of patient samples “Golden Staph”)
o Sample analysis protocols In 1870
o Dosing guidelines  Burdon-Sanderson, UK, attracted by Pasteur’s germ
o Trained medical, nursing and scientific theory
staff In 1871
Limitations of Salvarsan and “Sulfa Drugs” (Sulfonamides)  reports ability of Penicillium mould (from fruit &
 Narrow “spectrum 範圍 of action” e.g. didn’t kill a jam) to stop bacterial growth
big enough range of bacterial species  Joseph Lister [1827-1912] (found antiseptic 防腐
劑 properties of phenol)
o Observed curative 有療效的 properties of
Penicillium-soaked dressings on infected
wounds
Alexander Fleming and the Discovery of Penicillin
 Born in 1881 in Scotland
 Watched soldiers die of infected wounds in
Medical Corps 美國陸軍醫療部 in WW1
 Noted failure of antiseptics to cure internal
infections
In 1928
 Director, Inoculation Lab, St Mary’s Hospital,
London
 Worked on antibacterial properties of human nasal
secretions (lysozyme)
A Fluke 偶然 Observation by Fleming
In July 1928
 Fleming took 2-week vacation
PHAR1101 Week7 L1 – Penicillin (由青黴菌中提煉出的抗生素, 可以干擾細菌的細胞壁的合成- 有殺菌作用, 用於治療尿道炎、中耳炎、鼻竇
炎、支氣管炎、口腔感染和肺炎) & the Golden Age of Antibiotics
 Left used agar plates (Streptococcal cultures) on
lab bench
 Unusual cold snap (growth advantage to
Penicillium mould contaminant)
 Returned to work and noticed inhibitory effect of
mould on bacterial growth
Fleming’s “Half-Hearted” Follow-Up
 Fleming made bright yellow filtered broth from
Penicillium notatum mould
o Very active against growing Staph cultures
as well as against other bacterial species
 Non-irritating if applied directly to tissue
o Safe if injected into healthy mice (not
Staph-infected mice)
 Couldn’t purify active chemical (penicillin unstable)
o No ones know how to do so this discovery
sit for13 years
The Abrasive 不友好的 Australian who had more effect
upon the world than any other
Howard Florey
 University of Adelaide-trained pathologist
 Professor of Pathology, Oxford
In 1937
 He hired Ernst Chain (talented biochemist)
 Chain overcame penicillin instability & extraction
problems
o Prepared penicillin as a stable lyophilised
salt at pH 5-8
In March 1940
 Florey has stunning 令人驚嘆的 success in first
animal tests
In February 1941 - The First Human Tests
 43 years old policeman with invasive Strep 鏈球菌
and Staph 金黃色葡萄球菌 infections
o 200 mg penicillin (drip) + 100 mg every 3
hrs
 24 hr, strong recovery but drug ran out after 3 days
 Administered recycled penicillin (from urine)
o Good response but death after drug ran
out
 Subsequent patient also died (girl) before success
with 2 patients
In 1940-41 – Uncle Sam to the Rescue
 Two papers published in The Lancet
 British drug companies unable to help (war
pressures)
 US$5000 grant from Rockefeller foundation
allowed travel to US
 Links with US Dept Agriculture researchers
(isolated high yield Penicillium strain)
 Consortium with US companies e.g. Pfizer, Squibb,
Eli Lilly, Abbott, Merck, etc.
o Perfected large scale deep vat growth of
Penicillium mould
In November 1942 – An Unplanned Test (USA)
 Over 500 deaths in tragic night club fire (Boston)
 220 survivors secretly treated with penicillin
 US Military amazed by drug’s effectiveness
 By 1944, monthly US production > 130 billion units
 Sufficient for Allied troops at Normandy invasion
(1944)
The Race to Make Penicillin
In 1943
 Both US and UK teams made drug crystals but
found they were working with different
“penicillins” (wrong structures)
 Many variants soon isolated from Penicillium
broths
 WW2: > 1000 scientists, 39 unis & companies tried
to make drug synthetically (most gave up)
In 1945
 Dorothy Hodgkin (UK) solved unusual β–lactam
structure (X-ray crystallography)
In 1957
 John Sheehan (USA) – first “total synthesis”
The Semi-Synthetic Penicillins
 Chemists soon made a wide range of penicillin
family members
 Some common examples include:
PHAR1101 Week7 L1 – Penicillin (由青黴菌中提煉出的抗生素, 可以干擾細菌的細胞壁的合成- 有殺菌作用, 用於治療尿道炎、中耳炎、鼻竇
炎、支氣管炎、口腔感染和肺炎) & the Golden Age of Antibiotics

 In pharmaceutical science, people realised the

phenomenon of interspecies competition
between soil microbes the origins of many
antibiotics (protection and predation)
How Does It Work? o Because soil contains very high densities
Jack Strominger of bacteria, then perhaps we should start
 Professor of Pharmacology, Uni of Wisconsin screening soil bacteria to see if we can
(USA) identify any promising drugs that are by-
 Studied bacterial cell wall + around 30 enzymes products of this chemical warfare
needed to make it competition
Origins of some classic antibiotics
 Identified peptidoglycan– chains of aminosugars
cross-linked by small peptides
In 1965
 Famous paper on inhibitory effect of penicillin on
cell wall synthesis in bacteria
 Penicillin = CWSI – cell wall synthesis inhibitor
 Cell wall - semi-rigid but dynamic structure which
helps bacteria maintain their shape
o Identified transpeptidase as main target
Penicillin blocks a key “Crosslinking” step in Bacterial
Peptidoglycan Production for cell wall Antibiotics: The Big Picture
 It has been suggested that during the 20th
century, antibiotics have been responsible for a
ten-year increase in lifespan because of their
ability to diminish the threat of premature death
through bacterial infection
 This is compared to a two-year increase in lifespan
if all cancers were curable
Drug Resistance
 Loss of effectiveness common for most if not all
antibiotics
 Bacteria can develop resistance spontaneously
 Or acquire via plasmids
The “Golden Age” of Antibiotic Discovery  Often share multiple resistance mechanisms via
 After WW2, need for secrecy concerning penicillin plasmids
ended
 Allowed commercial development of many new
drug classes
 Widespread, global screening of microbes for new
drugs e.g. derived from soil samples
o Of all antibiotics discovered from 1945 to
1978, 55% came from the genus
Streptomyces (soil)
 Many lethal diseases receded e.g. pneumonia,
syphilis, gonorrhoea, diphtheria, scarlet fever,
childbirth infections, etc.
PHAR1101 Week7 L1 – Penicillin (由青黴菌中提煉出的抗生素, 可以干擾細菌的細胞壁的合成- 有殺菌作用, 用於治療尿道炎、中耳炎、鼻竇
炎、支氣管炎、口腔感染和肺炎) & the Golden Age of Antibiotics

The Economic Dimension of the Problem

 Drug companies need to receive adequate returns
on their antibiotic investments
o Recover huge discovery and development
costs
o Can’t do this is drugs quickly lose
effectiveness due to resistance
The Visible Impact of Multiple Drug Resistance (MDR)
 We need cheaper ways of discovering and testing
antibiotics in humans

 Clear zones around drug-containing disks on

bacterial cultures grown on agar plates are
indicative of sensitivity to the drug - drug stops
growth or kills bugs
 Lack of clear zones shows MDR bacteria are
resistant to the respective antibiotics
Consequences of Drug Resistance for Humanity
 The Golden Age is over
 More antibiotics are losing effectiveness
o ADR: Antibiotic Drug Resistance
o O’Neill Report (UK, 2016) – predicted 10
million deaths p.a. due to ADR by 2050
 Increases cost of treating patients with resistant
strains c.f. sensitive strains
 Resorting to nastier drugs
 “Multidrug resistant [MDR] strains” are growing
problem (hospitals) A Shrinking toolbox of new antibiotics since 1980
o Especially in low- and middle-income
countries (LMICs)
 Need to be smarter with existing drugs
Complexities of Antibiotic Drug Resistance in LMICs
PHAR1101 Week7 L1 – Penicillin (由青黴菌中提煉出的抗生素, 可以干擾細菌的細胞壁的合成- 有殺菌作用, 用於治療尿道炎、中耳炎、鼻竇
炎、支氣管炎、口腔感染和肺炎) & the Golden Age of Antibiotics

Good news 1 : European resurgence 復興 with improved

antibiotic stewardship 抗生素使用 in hospitals

Good news 2: Glimmers of hope 一线希望 on research

front from microbial genomics
 Modern high-throughput drug discovery methods
haven’t worked well for antibiotic discovery e.g.
GSK, A-Z, etc
 Microbe genome sequencing studies on drug
making bugs
o Only a low fraction (<25%) of drug-
making gene pathways are turned on in
lab-cultured soil microbes
o Can we switch dormant genes on to make
novel antibiotics?
 Ongoing research endeavour in many
microbiology labs
o Exploration of novel hard to reach
microbial niches