Title: Course: Scientific Discovery
1- Course Scientific Discovery
- Instructor Dr. Alexandra Vankley
- Presentation By Sri Ram
- 10/21/03
2STORY OF PENCILLIN
3Interesting Facts
- The wonder drug
- The first antibiotic popularly known
- The first antibiotic produced and used on a
massive scale - The pioneer of chemotherapy
4The 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
5Discovery of Penicillin
6Background
- Works of Louis Pasteur and Robert Koch, helped
elucidate the connection between infectious
diseases and the invasion of the body by bacteria
and other micro-organisms .
7Microbial Discoveries
8Related 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 .
9Related 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.
10Related 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.
11Sir Alexander Fleming
12Sir Alexander Fleming
- Born on August 6th,1881 at Lochfield, Scotland in
a farmers 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.
13Sir Alexander Fleming
- 1895 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.
14Sir Alexander Fleming
- Later encouraged by his brother and supported by
the money he received from an Uncles 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
15Sir 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.
16Sir 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.Marys.
- 1914-1918- Served as Captain in W W 1
-
1906 he joined the staff of the
17Sir 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.
18- 1948-Elected the Emeritus Professor of
Bacteriology, University of London. - 1951-54-He was Rector of Edinburgh University.
- Died on March 11th,1955
19Flemings 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.
20Ehrlichs Salvarsan
- In 1909 German chemist-physician Paul Ehrlich
developed a chemical treatment for syphilis. - salvarsan - "that which saves by arsenic".
- Ehrlichs 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.
21Antiseptics 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.
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23Lysozyme Discovery
- Fleming looked for -a chemical like salvarsan,
that could help fight microbe infections. - After war in 1920, back in St.Marys 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.
24Lysozymes Activity
25Lysozyme
- Lysozyme, in its natural state, seemed to be more
effective against harmless airborne bacteria than
against disease-causing bacteria. - And attempts to concentrate it, proved
unsuccessful. - Fleming continued his research of finding a
better and less toxic antiseptic and
antibacterial.
26The 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.
27Fleming at his usual work
28The 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.
29The Halo of Pencillin
30The culture-plate
31How 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.
32Flemings 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
bacteriaas in his lysozyme studies.
33Penicillin and Lysozyme Activity
34Flemings 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.
35Penicillin 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.
36Penicillin 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.
37Penicillin 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.
38Flemings 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). -
39Selective Inhibition
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41Penicillin 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.
42Paper on Penicillin
- Little notice was taken by the scientific
community of his paper published in the British
Journal of Experimental Pathology (June 1929).
43Explanation 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.
44Fleming-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.
45End of the 1st half
46The 2nd half
-
- The discovery and realization of the
chemotherapeutic potential of Penicillin
47Sir Howard Walter Florey
48Sir 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).
49Sir 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.
50Sir 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
51Sir 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.
52Ernst Boris Chain
53Ernst 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. -
54Ernst 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.
55Ernst 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
56Floreys 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.
57Florey 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 substratethe location on
bacteria to which it usually attaches.
58Future 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.
59Chains 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.
60Work 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
61Work 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.
62Isolating 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.
63Purification Difficulty
- Gallons and gallons of mold broth were used to
produce an amount just large enough to cover a
fingernail!
64First 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.
65Florey Impressed
- Florey directed that the antibacterial
properties of penicillin in mice be testedthe
step that Fleming had not taken.
66Heatleys 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.
67First 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.
68First Therapeutic Test
Mice Injected with Bacteria
with Penicillin
without Penicillin
69The 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!"
70The Miracle
The Mice injected with Penicillin survive !
71Publication
- 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.
72The First Trial on Humans
- The first patient was a young woman volunteer
with non-treatable cancer (January 1941) . - She showed an alarming reactiontrembling and
sharply rising fever. - With paper chromatography- Abraham was able to
separate out the impurities -showed that they
caused the adverse reaction.
73The 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.
74The 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.
75Efforts to Mass Produce
- Increasing production - overriding importance.
- Penicillium mold requires air to grow-
surface-cultured in regular laboratory flasks.
All manner of vessels used- hospital bedpans,
made-to-order ceramic pots,etc
76Industrial Production
- Florey approached various British pharmaceutical
firms. - British pharmaceutical firms-committed to
manufacturing other drugs needed for military and
civilian populations-devastated by enemy
bombardment.
77Entry 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.
78Joint Action
- Floreys 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 .
79Joint Action
- At the height of the program- the
British-American penicillin effort-thousands of
people and some thirty-five institutions
university chemistry and physics departments,
government agencies, research foundations, and
pharmaceutical companies.
80Only 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.
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82Efforts 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.
83Efforts 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 .
84Penicillin 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
85Proposed Structures
Proposed oxazalone structure forpenicillin. The
oxazalone ringis shown in red.
Beta-lactam ring structure forpenicillin. The
beta-lactam ringis shown in red.
86Newer 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.
87Production 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.
88Awards time
- World War II ended and the Nobel Prizes in
physiology or medicine distributed to Fleming,
Florey, and Chain for their work on penicillin.
89The Nobel Prize in Physiology or Medicine 1945
"for the discovery of penicillin and its curative
effect in various infectious diseases"
90Penicillin-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
91Words 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.
92References
- www.nobel.se
- www.chemheritage.com
- www.hisortylearningsite.co.uk
- www.pbs.org
- www.time.com