Transcript Alexander Flemming (presented by Sri Ram)

Course: Scientific Discovery
Instructor: Dr. Alexandra Vankley
Presentation By: Sri Ram
10/21/03
STORY OF PENCILLIN
Interesting Facts
 The
wonder drug
 The first antibiotic popularly known
 The first antibiotic produced and used on a
massive scale
 The pioneer of chemotherapy
The 2 parts of the story
 a-the
discovery of penicillin as an
antibacterial and Alexander Fleming
 b-the discovery and realization of its
chemotherapeutic potential
Discovery of Penicillin
Background
 Works
of Louis Pasteur and Robert Koch,
helped elucidate the connection between
infectious diseases and the invasion of the
body by bacteria and other microorganisms .
Microbial Discoveries
Year
1880-82
Microbe
discovered
Typhoid
1883
Cholera
1884
Tetanus
1886
Pneumonia
1894
Plague
Related Events

In 1871, Joseph Lister ( antiseptic practice
surgery) - a mold in a sample of urine - inhibiting
bacterial growth.
 In 1875 John Tyndall - a species of Penicillium
caused some of his bacteria to burst.
 In 1877 Louis Pasteur and Jules Joubert airborne microorganisms could inhibit the growth
of anthrax bacilli .
Related Events contd.

Ernest Duchesne in 1897 focused on the
interaction between E. coli and Penicillium
glaucum .
 Inoculating mold and typhoid bacilli -prevented
contraction of typhoid in animals. But, he died of
T.B before he could complete his research.
 Experiments carried out by Emmerich and Loew
(1899) and later by Gratia and Dath and others
did not give any favorable results.
Related Events contd.
 Instead
various forms of vaccination and
serum treatment were evolved.
 The first Nobel Prize for Physiology or
Medicine in 1901 was given for serum
therapy for diphtheria.
 Human and animal bodies - produce
protective substances in the fight against
the invaders.
Sir Alexander Fleming
Sir Alexander Fleming
on August 6th,1881 at Lochfield,
Scotland in a farmer’s family .
 He had his early days spent more in the
farms but was sound in his fundamental
education.
 1895-The untimely death of his father and
the success of the medical practice of his
step-brother Tom had him relocated to
London.
 Born
Sir Alexander Fleming
–He attended the Polytechnic School
in Regent Street .
 1896-1900- He worked as a clerk in a
shipping firm.
 1900- In 1900, when the Boer War broke
out joined the Scottish regiment but
never saw the combat.
 1895
Sir Alexander Fleming
 Later
encouraged by his brother and
supported by the money he received from
an Uncle’s demise he looked towards a
medical career.
 1901-He won a scholarship to St. Mary's
Hospital Medical School, London
University, Paddington and joined it.
 He qualified with distinction and received
his degree in 1906
Sir Alexander Fleming

His switch to bacteriology was even more
surprising: if he took a position as a surgeon, he
would have to leave St. Mary's.
 The captain of St. Mary's rifle club knew that and
was desperate to improve his team. Knowing
that Fleming was a great shot he did all he could
to keep him at St. Mary's.
 He worked in the Inoculation Service and he
convinced Fleming to join his department in
order to work with its brilliant director -- and to
join the rifle club.
Sir Alexander Fleming

1906- Fleming joined the Inoculation Department as
medical bacteriologist under the direction of Sir
Almroth Wright.

He made the St. Mary's Hospital Medical School his
professional home for the rest of his life.

1914-Flemming became a lecturer at St.Mary’s.

1914-1918- Served as Captain in W W 1
Sir Alexander Fleming
 1918
-He returned to St.Mary's.
 1928- He was elected Professor of the
School in 1928
 1943-He was elected Fellow of Royal
Society.
 1944-Flemming was knighted.
 1945-Received his Nobel Prize.
 1948-Elected
the Emeritus Professor of
Bacteriology, University of London.
 1951-54-He was Rector of Edinburgh
University.
 Died on March 11th,1955
Fleming’s Ideology
 Wright
believed, as did Fleming, there
were substances in the human body that
could be used to fight infection, and
strengthening the immune system was
key.
 Hence, they believed more on vaccine
therapy rather than chemotherapy.
Ehrlich’s “Salvarsan”

In 1909 German chemist-physician Paul Ehrlich
developed a chemical treatment for syphilis.
 salvarsan - "that which saves by arsenic".
 Ehrlich’s samples were given to Fleming by
Wright and became efficient in its administration.
 He did so with the new and difficult technique of
intravenous injection. He soon developed such a
busy practice he got the nickname "Private 606.“
Antiseptics of that period

During World War I- wound-research
laboratory in Boulogne, France.
 Chemical antiseptics like carbolic acid
(phenol) do not sterilize jagged wounds;
rather, pus has its own antibacterial powers.
 He was able to show that chemical
antiseptics in dilutions harmless to bacteria
actually damage white blood corpuscles
(leukocytes)—the body's first line of defense.
Lysozyme Discovery

Fleming looked for -a chemical like salvarsan,
that could help fight microbe infections.
 After war in 1920, back in St.Mary’s Fleming
searched for an affective antiseptic.
 He discovered Lysozyme, in nasal mucus . Its
an enzyme found in many body fluid, like tears,
etc. It is a natural antibacterial not effective
against the stronger infectious agents.
Lysozyme’s Activity
Lysozyme
 Lysozyme,
in its natural state, seemed to
be more effective against harmless
airborne bacteria than against diseasecausing bacteria.
 And attempts to concentrate it, proved
unsuccessful.
 Fleming continued his research of finding
a better and less toxic antiseptic and
antibacterial.
The Chanced Discovery
 Fleming's
legendary discovery of penicillin
occurred in 1928, while he was
investigating staphylococcus, a common
bacteria then caused diseases ranging
from boils to disastrous infections.
Fleming at his usual work
The Halo of Pencillin
He left a culture plate smeared with
Staphylococcus bacteria on his lab
bench while he went on a two-week
holiday.
 When he returned, he noticed a clear
halo surrounding the yellow-green
growth of a mold that had accidentally
contaminated the plate.

The Halo of Pencillin
The culture-plate
How This Happened
 Luckily,
Fleming had not stored his
culture in a warm incubator. London
was then hit by a cold spell, giving the
mold a chance to grow.
 Later, as the temperature rose, the
Staphylococcus bacteria grew,
covering the entire plate--except for the
area surrounding the moldy
contaminant.
Fleming’s Deduction
 Fleming
correctly deduced -mold must
have released a substance - inhibited
the growth of the bacteria.
 He was never clear on his
observations. The evidence of the first
culture, which he photographed
indicated that Fleming observed lysis,
the weakening and destruction of
bacteria—as in his lysozyme studies.
Penicillin and Lysozyme Activity
Fleming’s Deduction
 But
sometimes Fleming described it
inhibition, or prevention of bacterial evidenced by a clear zone surrounding
the mold.
 Although these two effects occur under
quite different conditions, Fleming
probably forgot which observation
came first.
Penicillin Identified

He discovered that the antibacterial
substance was not produced by all molds,
only by strain of Penicillium notatum).
 Although he could not isolate it, he named
the active substance “penicillin.”
 He studied methods of producing the impure
product and determined its stability at
different temperatures and over various
lengths of time.
Penicillin is Non-Toxic
 Fleming
found that penicillin was not toxic
to animals and that it did not harm white
blood cells (leucocytes) by injecting
healthy mice extract from the mold growth.
 Previous, observations of such injections
proved toxic and penicillin was an
exception.
Penicillin Properties
 Penicillin
would not be absorbed if
taken orally.
 Penicillin taken by injection - excreted
in the urine in a matter of hours-well
before it could have its effects.
 Moreover, the unstable penicillin was
never available for clinical testing.
Fleming’s ways of using Penicillin
 For
Fleming, penicillin's therapeutic
potential - topical antisepsis.
 Fleming did continue to use in
bacteriology. Penicillin suppressed the
growth of certain bacterial species,
allowing one to selectively culture
certain others (such as those causing
influenza, acne and whooping cough).
Selective Inhibition
Penicillin in vaccine production
 Penicillin
became a valuable in
manufacture of vaccines.
 The penicillin was crude--good enough
for Fleming's purpose, but hardly
strong enough to destroy a serious
human infection.
Paper on Penicillin
Little notice was taken by the scientific
community of his paper published in
the British Journal of Experimental
Pathology (June 1929).
Explanation for Failure
 His
belief - cure comes from within the
body itself rather than from an external
chemical agent.
 Difficulties -he had experienced in
isolating and stabilizing penicillin,
producing sufficient quantities for
clinical trials- prevented him from
realizing the full fruits of his research.
Fleming-After Penicillin

Fleming had turned his research to other
chemical antibacterials, the sulphonamides
 He kept producing and supplying his sample
of penicillin to other labs and researchers for
various studies and experiments.
 For more than a decade or so no progress
was made in the discovery of penicillin.
End of the 1st half
The 2nd half
The discovery and realization of the
chemotherapeutic potential of Penicillin
Sir Howard Walter Florey
Sir Howard Walter Florey
 Born
on September 24, 1898, at
Adelaide, South Australia .
 He graduated in M.B., B.S. in 1921from
Adelaide University.
 He was awarded a Rhodes Scholarship
to Magdalene College, Oxford, leading
to the degrees of B.Sc. and M.A. (1924).
Sir Howard Walter Florey

In 1925 he visited the United States on a
Rockefeller Travelling Fellowship for a year,
 Returned in 1926 to a Fellowship at Gonville
and Caius College, Cambridge, received his
Ph.D. in 1927.
 In 1931 he succeeded to the Joseph Hunter
Chair of Pathology at the University of
Sheffield.
Sir Howard Walter Florey
 Leaving
Sheffield in 1935 he became
Professor of Pathology and a Fellow of
Lincoln College, Oxford.
 1936-He was made Director to the Sir
William Dunn School of Pathology.
 He was made an Honorary Fellow of
Gonville and Caius College, Cambridge
in 1946
Sir Howard Walter Florey
 Honorary
Fellow of Magdalen College,
Oxford in 1952.
 In 1962 he was made Provost of The
Queen's College, Oxford
 In 1944 he was created a Knight
Bachelor.
 In 1945 was awarded the Nobel Prize.
 Dr. Florey died in 1968.
Ernst Boris Chain
Ernst Boris Chain

Born on June 19, 1906, in Berlin.
 He next attended the Friedrich-Wilhelm
University, Berlin, where he graduated in
chemistry in 1930.
 He worked for three years at the Charité
Hospital, Berlin, on enzyme research.
 In 1933, after the access to power of the Nazi
regime in Germany, he emigrated to England.
Ernst Boris Chain


He first worked on phospholipids at the
School of Biochemistry, Cambridge, under
the direction of Sir Frederick Gowland
Hopkins .
In 1935 he was invited to Oxford University
where he worked in the Sir William Dunn
School of Pathology.
 in 1936, he was made demonstrator and
lecturer in chemical pathology.
Ernst Boris Chain
 In
1948 he was appointed Scientific
Director of the International Research
Centre for Chemical Microbiology at the
Istituto Superiore di Sanita, Rome.
 He became Professor of Biochemistry
at Imperial College, University of
London, in 1961
 1945-Awarded Nobel prize
 Dr.Chain died in 1979
Florey’s Team
 Florey
recruited -a interdisciplinary
group of scientists- to study
pathological evidence of disease and
physiological processes by which
those symptoms arose, traced to the
chemical and even the molecular level.
Florey joined by Chain
 Among
his first hires was the
biochemist Ernst Boris Chain.
 With Chain , one of the projects
pursued was the crystallization of
lysozyme and the characterization of its
substrate—the location on bacteria to
which it usually attaches.
Future Plans
 In
1938, while the lysozyme research
was concluding, Florey and Chain
decided to study selected antibacterial
substances produced by certain
microorganisms. They thought these
substances were all enzymes like
lysozyme.
Chain’s suggestion
 This
process was greatly facilitated by
Chain's near-photographic recall of the
many scientific papers he had read,
including Fleming's 1929 paper on
penicillin.
Work on Penicillin Begins
 They
originally chose substances from
three organisms: pyocyanase, a topical
antibacterial, from Bacillus
pyocyaneus; extracts from certain
organisms in the soil called
actinomycetes; and penicillin
Work on Penicillin
 While
Florey and Chain were
assembling grants and funds, work was
begun on penicillin.
 Fortuitously, there was already a
penicillin culture at the William Dunn
School
 The research program rapidly narrowed
its focus to penicillin alone.
Isolating Pure Penicillin
 Chain,
along with another chemist,
E.P.Abraham -technique for purifying
and concentrating penicillin.
 The key- pH of the “juice,” the
sample's temperature, freeze-drying it.
 Later improved on by Norman Heatley
and other scientists.
Purification Difficulty
 Gallons
and gallons of mold broth were
used to produce an amount just large
enough to cover a fingernail!
First Toxicity Test
 In
March 1940 Chain injected mice with
a sample of the penicillin extracted.
 Far higher dosage than Fleming's - the
mice survived apparently unharmed.
 The more-concentrated penicillin had
passed its first toxicity test.
Florey Impressed
 Florey
directed that the antibacterial
properties of penicillin in mice be
tested—the step that Fleming had not
taken.
Heatley’s Contributions
 Norman
Heatley -technical inventions
to produce penicillin on a larger scale.
 Heatley- contributed a lot to the
purification process and different
methods of growing mould in various
containers.
First Therapeutic Test
 In
May 1940, the team had been able to
produce enough penicillin to test on
infected animals .
 Eight mice were infected with a deadly
dose of streptococci bacteria each. One
hour later, four of them were injected
with penicillin and four mice were left
without treatment.
First Therapeutic Test
Mice Injected with Bacteria
with Penicillin
without Penicillin
The Miracle
 The
four mice that hadn't been injected
with penicillin started to show signs of
illness and later died.
 The four mice treated with penicillin
remained fine!
 Hearing this Howard Florey exclaimed,
"It looks like a miracle!"
The Miracle
The Mice injected with Penicillin survive !
Publication
 On
24 August 1940 Florey and Chain
reported their findings in the Lancet;
 By
then World War II had already
engulfed Europe, calling for finding
means of combating the diseases and
infections of war to hold the advantage.
The First Trial on Humans

The first patient was a young woman
volunteer with non-treatable cancer (January
1941) .
 She showed an alarming reaction—trembling
and sharply rising fever.
 With paper chromatography- Abraham was
able to separate out the impurities -showed
that they caused the adverse reaction.
The First Clinical Trial
 On
12 February 1941 a policeman with
an invasive infection - first patient with
an infection to be treated with
penicillin.
 The dosages and the length of
treatment required were being worked
out by just trials.
The First Clinical Trial
 First
improved and then relapsed.
 The penicillin supply ran out- even
retrieving penicillin from the man's own
urine- failed to save him.
 Florey vowed that from then on he
would always have enough penicillin to
complete a treatment.
Efforts to Mass Produce
 Increasing
production - overriding
importance.
 Penicillium mold requires air to growsurface-cultured in regular laboratory
flasks. All manner of vessels usedhospital bedpans, made-to-order
ceramic pots,etc
Industrial Production
 Florey
approached various British
pharmaceutical firms.
 British pharmaceutical firms-committed
to manufacturing other drugs needed
for military and civilian populationsdevastated by enemy bombardment.
Entry of USA
 To
obtain the assistance of the United
States- noncombatant- in increasing
production and furthering research,
Florey and Heatley flew across the
Atlantic in the beginning of July 1941.
Joint Action
 Florey’s
contacts helped him get to
have the production taken up by the
U.S Firms and Government agencies.
 Soon it was decided to have an
expedite unified action on penicillin .
Joint Action
 At
the height of the program- the
British-American penicillin effortthousands of people and some thirtyfive institutions: university chemistry
and physics departments, government
agencies, research foundations, and
pharmaceutical companies.
Only Life Saver in War
By D-day there was enough penicillin
on hand to treat every soldier who
needed it. By the end of World War II, it
had saved millions of lives.
Efforts to Study Penicillin
 Some
chemists -synthesize penicillin
from a few organic chemicals.
 Efforts to understand the molecular
structure of the penicillin.
 Reacting the substance with various
chemical reagents, which resulted in
products of known structure.
Efforts to Study Penicillin
 From
these bits of structural
information -deduce how the original
molecule was organized.
 At Oxford the problem of determining
penicillin's structure was given to
Chain, Abraham, and Robert Robinson,
a senior organic chemist .
Penicillin Structure
 By
fall 1943, groups working at Oxford
and at Merck had proposed two
different structures penicillin molecule.
 Chain and Abraham as well as by
Robert Burns Woodward at Harvard—
four-membered beta- lactam ring lay at
the heart of the penicillin molecule
 Robinson instead proposed a structure
based on oxazalone
Proposed Structures
Beta-lactam ring structure for
penicillin. The beta-lactam ring
is shown in red.
Proposed oxazalone structure for
penicillin. The oxazalone ring
is shown in red.
Newer Techniques to Study
 New
techniques for analyzing the
structure of organic molecules - X-ray
crystallography- practiced by Dorothy
Hodgkin
 In 1945 she was able confirmed Chain
and Abraham's deduction. This
evidence ran counter to Robinson's
proposed structure for penicillin.
Production by Fermentation only
 In
1957 John Sheehan created such a
synthesis, but fermentation - the
commercial production of penicillin and
related antibiotics.
 But the structural knowledge gained developing penicillin-like antibiotics
that were more effective, convenient to
give and had fewer side effects.
Awards time
 World
War II ended and the Nobel
Prizes in physiology or medicine
distributed to Fleming, Florey, and
Chain for their work on penicillin.
The Nobel Prize in Physiology or Medicine 1945
"for the discovery of penicillin and its curative effect in various infectious diseases"
Penicillin-The master Drug
 Pneumonia,
syphilis, gonorrhea,
diphtheria, scarlet fever and many
wound and childbirth infections that
once killed indiscriminately suddenly
became treatable.
 Deaths caused by bacterial infections
plummeted
Words of Caution
 By
this time, even Fleming was aware
that penicillin had an Achilles' heel.
 He wrote in 1946 that "the
administration of too small doses ...
leads to the production of resistant
strains of bacteria." It's a problem that
plagues us to this day.
References
 www.nobel.se
 www.chemheritage.com
 www.hisortylearningsite.co.uk
 www.pbs.org
 www.time.com