Dr. SubbaRow was instrumental in setting up a
number of graduate fellowships in pharmacy schools
and different schools throughout the country, and
one of these was established at the University of
Minnesota through Professor Charles 0. Wilson who
was my professor. I was the first graduate student
at Minnesota to have this Lederle fellowship, and
from that developed the contacts with Dr.
SubbaRow, which resulted in my coming here for an
interview for a job.
I worked
on the fellowship for 1 year - this was for a
Ph.D. The only other one from Minnesota is Dr.
Martel who is in my group quite new. My wife was
hired by Dr. SubbaRow too about the same time I
was - she's a Ph.D. bacteriologist. She's from
Yale. Her maiden name is Jane Hill. I was
acquainted with SubbaRow's work only because I was
working on a Lederle fellowship. I never did any
work that was connected with the work that he had
done. The impression I got from my interview with
him was sort of an odd one.
He
asked me very few questions and made up his mind
in a few minutes, and made me an offer of a job
right away. I think I embarrassed him a little
bit. He asked me how much money I expected and I
named a figure, and I think it was less than he
expected to offer me. He explained that it wasn't
going to cost me more to live here than I think
and he said we're going to offer you such and
such, which was more than the figure I'd
mentioned. His office was exactly where
I am now - the blackboard was his. I have
a little shelf where the telephone is, which I
think was his also. His secretary was Barbara
Merriam. The men working with me then were Dr.
Bohonos, Hutchings, Stokstad - those 3 were the
main ones I was actually working with. Stokstad
was working on folic acid. Jukes was here, but he
hadn't been here very long. He was in Sales
Education. I worked for actually 2 months on a
separate problem called the sprue factor which was
related to folic acid and later became part of it.
There was a material that Dr.
Bohonos had isolated from liver which was known as
the sprue factor and it was thought at the time to
have something to do with sprue - perhaps the lack
of it causes sprue. The compound was identified in
a mater of a few months - it was identified as a
known compound and then we no longer worked on
that. It was identified as one of the B vitamins
which had nothing to do with sprue and nothing to
do with folic acid either. After that I started
working on folic acid problem. The first few
months I was helping in isolation - the isolation
had already been worked out by other people -
Hutchings, Bohonos and Stokstad but there was a
problem of continual supply and I was doing this.
Yes, it was made from fermentation. I was doing
the last few steps in the isolation. I was also
doing chemical work on the pteridines at the time
� we thought pteridines were somehow related to
folic acid - there were some known pteridines in
the literature at the time and I was spending some
time making these - they are related as there is a
pteridine ring in folic acid.
The work I was doing in the first year was mainly
these two things - doing the last steps in the
isolation fermentation of folic acid, and making a
few known pteridines for practise and seeing what
they were like in preparation for determining the
structure of folic acid. After 6 or 8 months we
started to study the degradation and try to
determine the structure of folic acid. Most of the
degradation work was done by Hutchings and
Stokstad. It was at about this time Dr. Mowat came
on the problem. He and I began to work sort of
together in synthesizing things that we thought
were closely related to degradation products. In
other words, Hutchings and Stokstad were doing the
actual degradation work on folic acid and Mowat
and Boothe were doing the synthesis of the
degradation product that they were getting out.
Waller came after about 1 year and Mowat and I had
already done some work on synthesizing these
pteridines. Waller also began to work in this area
and Angier came about that time also. I think
Waller made the first successful synthesis.
Of course, SubbaRow was overseeing all
of this. He wasn't doing any laboratory work
himself. He was constantly in the laboratories and
my laboratory was right next to his and SubbaRow
was in and out all the time. He knew day to day
what we were doing. This was a natural product
problem that he could see a solution coming up and
he was very interested in finishing this problem.
This was a new vitamin and new growth factor and
he wanted us to be first in this. He would walk in
and pace around and talk and he would ask us how
are things.
He would often
suggest, why don't we try this, or how did this go
- he vas intimately interested in the results each
day. He was a good supervisor - sometimes it did
seem he came too often - I don't think anyone
resented this at the time. There was a feeling
that this was a very important problem and there
was a feeling that you were being pushed some.
There was a feeling that Dr. SubbaRow was anxious
and that the people above him were anxious too -
there was a feeling of urgency, of getting the
thing done. If we weren't first in this there was
something wrong because we had a good head start -
we had a lot of people working on it and we felt
we should be first. It gave us all very good
experience on working on many things - natural
products, the difficulties involved, good
experience in how to attack other natural product
problems.
The name of the
degradation product was
2-amino-4-hydroxypteridine-6-carboxilic acid. This
was always known in our laboratory as fraction l
B. That was the first degradation product that
gave a clue to the structure of the whole thing.
And there was also 2-amino-4-hydoxy-6
methylpteridine. Xanthopterin does show some
activity and I think it must be that the species -
fish or animals - converted it somehow
biologically. Maybe they can use it as a precursor
to make folic acid.
Dr. Waller
was the first one to synthesize folic acid. Later,
I synthesized it also by a couple of other methods
and later synthesized the fermentation folic acid.
I was the first one to synthesize that - that has
3 glutamic acids in place of 1 and the main
problem there was that it added on the 2 extra
glutamics. Lederle did sell for a number of years
the fermentation folic acid that was known as
Teropterin. We thought we were hot on the trail
for a cure for cancer at the time and that was why
were trying so hard to synthesize the fermentation
folic acid so that we would have enough of it to
really try. We didn�t have enough from
fermentation sources and the synthesis did provide
rather large quantities. We made quite a number of
kilos in the very early years. That was clinically
tried in cancer. Liver folic acid has been
synthesized by a large number of methods. Each of
us in the group - there were 5 of us - each of us
synthesized it by a different method.
I don't remember the day Dr. Waller synthesized
folic acid - he was not in my lab at the time and
I'm a little hazy about that. No one knew it until
it was assayed because the material that he got
out was not pure. You couldn't even tell that it
had folic acid in it until it was biologically
assayed - this took a few days and then no one
believed it so it had to be repeated a few times.
The discoveries never come all in one day - it's
sort of a gradual thing.
It
was made on a large scale by that process for
quite a long time. Several years later I think
there was a different one substituted rather than
the first original shotgun method but that was
used for quite a number of years and was as cheap
as any. The way this interest in cancer developed
there was a group in a hospital in New York City
that had a cancer testing program in which they
transplanted cancer into mice and then tested
compounds to see which ones prevented the growth.
This group tested fermentation folic acid and they
claimed it was active in cancer - they claimed it
prevented the growth of these transplanted tumours
in the mice and this vas the first connection we
had between folic acid and cancer. This then
stimulated the work toward synthesizing the
fermentation folic so that we could have enough to
test it.
I can vaguely recall
Dr. SubbaRow coming and talking to Mowat and
myself. Mowat was the first one who started to
work on this and then I joined him a few months
later. There are several ways you can put 3
glutamic acids together and it was by chance that
Mowat selected certain ones that were the wrong
ones and I selected the ones that were the right
ones. This took a year or year-and-a-half. Jukes
was sort of on the sidelines - an advisor to
Stokstad but he was not directly connected with
the synthesis of folic acid at all. Stokstad also
worked on the mode of action and the citrovarum
factor - but this came later. I was not involved
with citrovarum at all - the synthesis of the
pteridin-6-carboxilic acid was a key to the good
bit of the structure. We worked on the synthesis
of that for quite some time and this was first
synthesized by Mowat.
The
problem involved with pteridine was there was no
good method for purification. When we synthesized
the product - it was very difficult to purify. I
think this WAS the main stumbling block in the
folic acid work and pteridine was difficulty in
purification. We had meetings but I never remember
any minutes or records kept of meetings. We had
many verbal meetings during which we discussed
these things and used blackboards, but I don't
know of any records of meetings. I think SubbaRow
often wrote on the blackboard, but I don't
remember that he ever wrote on a sheet of paper.
The biggest problem of solving the problem before
I came was the discovery of the way to ferment.
Liver folic acid was extremely hard to
isolate - this was Dr. Hutchings discovery - the
method of micro organism that would make a folic
acid that we could isolate. The fermentation
method was a practical way whereby we could get a
half a gram a week whereas from liver it was much
less than that but I can't quote any amounts.
Before the folic acid was synthesized someone did
a cost estimate by making by fermentation and it
was possible that you could make and sell this by
the fermentation procedure. Of course once it was
synthesized it was much cheaper to make it that
way.
We cooperated with the
group from Bound Brook and I'm not too sure when
it started - in 1944 or sometime. There were 8
people from Bound Brook and 8 from Pearl River
that were cooperating and we did have meetings
quite regularly. We had meetings quite often in
New York City - we'd meet in the New York office
of Cyanamid. I remember one time we were scheduled
to meet there for a meeting. Dr. SubbaRow always
drove us and we got there and no one came from
Bound Brook. Dr. SubbaRow called up and they said
they didn't know there was a meeting so we had
dinner and drove back home. He'd always take us to
dinner when we had one of these meetings.
Dr. Northey was the supervisor of these
people from Bound Brook. I would say that Dr.
SubbaRow contributed more than Dr. Northey. Dr.
Northey was at his best in planning the
manufacture of something. He was very practical
minded - he knew how big a plant it would take to
manufacture so many kilos of folic acid per month.
In the ideas for research purposes Dr. SubbaRow
had a better grasp of that.
I
don't know of any real disagreement among the
group. I think for the number of people involved
and the number of individualists involved it was
remarkable that we didn't have more disagreements
than we had. In addition to these meetings we also
telephoned quite regularly and we had meetings
about once a month. I consider that Dr. SubbaRow
was a direct contributor to the research. He
contributed as much perhaps more than others. I
don't think anyone of us considered him a
supervisor but an actual contributor as much as
anyone to the problem. Folic acid is certainly one
of the required vitamins and I think it's probably
as important as most of the others. It has this
one difficulty of disguising the symptoms without
curing pernicious anaemia but this doesn't make it
any less important on the basis of nutrition.
Nutritionally it's still a very important vitamin.
SubbaRow wasn't a very good
driver and we were always a little worried when
there was 6 of us in his car. The trip we made to
Bound Brook and brought back the first bottle of
folic acid they made - a large quantity and this
was not too long after the first synthesis. This
was very impressive to see perhaps a kilo of folic
acid. They had scaled it whereby they could make a
kilo. SubbaRow was more of a biochemist than an
organic chemist although he had a good
understanding of organic chemistry. I think this
cooperation with Bound Brook began because
SubbaRow was short of chemists and they wanted to
put a lot of effort in it and this was the best
way to do it quickly.
But I
don't think he intended to amalgamate the two labs
together. In the period just after the war until
1948 he had continually added chemists and the
number increased but I don�t know how they
compared with Bound Brook and Stamford. I think he
felt that the person with background and training
was not really the important thing - they could
learn any of these. If an organic chemist had an
interest in biochemistry he could learn any of
this material. I don't think he felt if an organic
chemist came he didn't have to stay at that. I
don't think he felt you had to stay at the thing
you majored in college.
I
think there was a change in SubbaRow's personality
even before I came - he was much more demanding.
Everyone felt they must work long hours. Even
before I came here in 1943 most people worked,
some at night, and in the years later they told me
everyone felt they must do this. But this was sort
of sloughing off when I came, and after I came
people no longer felt they were compelled to work
at night. He felt that this wasn't the only thing
you had to do.
At the time I
came he was investigating many hobbies. He was
looking for things for his spare time, and this
was an indication that he no longer felt his work
was the only thing. He had taken up horseback
riding and golf and airplane. There were many
hobbies he took up - some he discarded very
quickly, some he continued with. I felt that he no
longer had this urge to work continually so people
felt that they didn't have to work as hard. He was
interested in many things in regard to medical
research - almost any subject he was interested in
and would have liked to work on. I can remember
discussing with him the problem of polio before it
was ever worked on here at all and he was
interested in cancer research. He was the one who
really got it started here - our connection
between folic acid and cancer and his interest in
it. I think his interest stemmed from . . . from
New York. I think his interest stemmed from that.
I don't think there was any fundamental concept. I
think it was just a chance observation by another
group and his taking advantage of this.
I think he had a broad interest in any
medical, biological problem but I don't think - I
wouldn't give him credit for having fundamental
idea of testing anti-metabolites in cancer. I
think this was sort of a gradual thing that came
along. I don't think his interest in sprue was an
obsession but he had an abiding interest in it. He
did have more of a personal interest in that than
the rest of us. I only knew it by name - I didn't
even know the disease existed until I came here. I
think his interest was as a medical man who had
seen examples of this - seen the effects of it on
people - maybe his family, I don't know.
Merck announced the structure of B12
before his death. I was never exposed to the B12
work but I knew very well the man who was doing
the work - Otto Wieland. He worked for years on
fractionating liver and having it tested in
pernicious anaemia patients. But that was hopeless
because there just weren't enough patients and the
test wasn't good enough. Otto used to complain
about the fact that he couldn't tell what he was
doing. One test would be positive and the next
would be negative, but I never heard him complain
that Dr. SubbaRow wasn't supporting his program.
Dr. SubbaRow wanted to be known as an
inventive man in the world of medicine. I had the
impression that he certainly wanted his name
known. He wanted to be remembered as an important
man in medicine - his contributions to be known.
But I don't known of a particular incidence where
he indicated this. He wasn�t considered in the
same class as a Nobel prize winner - maybe someday
he would have been. I think he had the normal urge
to be known by his fellow scientists as much as
many of us - perhaps more than many of us. I don't
think it was his choice if he was not given proper
recognition. Had he lived another 5 years, I think
he would have gotten the publicity that Dr. Duggar
did get. I think his name would have been
connected with Aureomycin in place of Duggar.
I didn't work on Aureomycin until 1950.
During the structure work I synthesized
degradation products. I was the first one here to
make tetracycline from Aureomycin - you remove the
chlorine from Aureomycin and make tetracycline. I
was the first one to do this here at Pearl River.
We later found that the Pfizer people had done it
before we did and the Pfizer people got the patent
on tetracycline. Lederle was the first to market
tetracycline and we were in a good competitive
position. For about a year or so I worked on
making analogues for folic acid and other
compounds to be tested. It was sort of a cancer
chemotherapy program at the time and I worked on
this until 1950 when I started to work on the
Aureomycin program.
I first
learned of his death in San Francisco 2 days
after. My home was on the West Coast in the state
of Washington. I was going to a meeting of the
American Pharmaceutical Association in San
Francisco to talk about Teropterin. I went to my
home in Washington first and then went from there
to San Francisco. It was there that I learned that
he had died.
I met Dr. Waller
there and he was the first one to tell me. It was
a shock. I probably saw him a week or two before I
left. I lived in the same building he did. I lived
in the apartment house where he lived and I used
to see him there quite often - coming and going. I
don't think I ever was in his apartment - maybe
once I was in his apartment I guess. I can
remember the people living under him complaining
he did a lot of pacing - he used to pace up and
down at night. I remember he used to buy crates of
oranges and eat them. This was a large part of his
diet - he ate tremendous amounts of oranges. The
majority of people who lived there were Lederle
people.
He was a complex
person - he had a much wide range of interests. He
was a nice person to work for - I enjoyed working
for him. He tended to be moody and did things
spontaneously - sometimes without too much
thought. He sometimes spoke out without giving
thought to the consequences and later sort of
regretted this. I think he did have likes and
dislikes - this is choosing people you expect to
run your research program. I think he did decide
certain people were better than other people. He
did have people he liked and I think he liked them
because they did a good job.
I
don't think he was unfair. He chose people as his
favourites but I don't think personalities had
anything to do with it. He was judging their
ability to do what he wanted. I often wonder how
he would have run a research group as it grew much
bigger. Years after he died it increased quite
rapidly and he was never one who could delegate
responsibility easily. He liked to keep everything
in his hands - whether he would have learned to
delegate responsibility to other people. If he had
tried to keep too tight a hold as the thing grew
he would have lost control entirely because no
person could keep the control he'd like on several
hundred people. He would have had to designate
people to be heads of departments and this was
something he had no experience with and didn't do
easily. He didn't like to delegate authority to
other people. He would delegate work to other
people but the person was responsible to him. He
wanted to know about it personally. He had a
prodigious memory for details - he would have
reached a point where he couldn't have done this
and I don't know how he would have reacted. I
think he had years of scientific usefulness yet. I
think if he would have learned to delegate
authority I think he would have done very well.
There were people who didn't like his methods and
quite a number of them left. Dr. Bohonos left a
year after I came - I don't think it was any great
feeling against SubbaRow though.
(Participating: Dr Edgar L Milford. Venue: Pearl
River, New York, USA. Date: 7 April 1965)
