Experiment Eleven (5 page)

By the beginning of World War Two, Merck was also producing vitamins. First came vitamin B1. Until the Merck chemists figured out how to synthesize the compound, tons of rice bran went into one end of Merck's Rahway plant, and fractions of an ounce of vitamin B1 came out the other end. Soon there was vitamin B2 for pellagra, vitamin B12 for anemia, vitamin C for colds, and vitamin A for eyesight. George Merck was also keeping a watchful eye on the Rutgers Department of Soil Microbiology. Like many other microbe researchers, Selman Waksman was experimenting with ways to use fungus fermentations to make citric acid, used in foods and soft drinks, and fumaric acid, used in dry cleaning.

At the beginning of 1939, Merck engaged Waksman as a consultant on
microbial fermentation, first at $100 a month and then $150. In the summer of that year, Merck funded a $150-a-month student fellowship specifically to find new ways of making citric acid. The program was successful, and the consultancy and the fellowship were renewed on an annual basis.

None of Waksman's students knew about his consultancies with Merck,
not even his deputy
, Bob Starkey. Some of the funds went into fellowships and stipends for his graduate students, some for “collaboration” between Merck and his laboratory, and some for “private consulting” between himself and Merck.

Toward the end of 1939, Merck expressed interest in hiring Waksman as a consultant on antibiotics, although in those days the word
antibiotic
was not in common usage. Merck called them “antibacterial
chemotherapeutic agents
.” One such agent was penicillin. Although Alexander Fleming had discovered penicillin's antibacterial powers in London in 1928, he had never found a method of storing, concentrating, or purifying it, and it had remained a laboratory curiosity.

American scientists had read about Fleming's discovery, and in 1933 a graduate student at Pennsylvania State College had studied Fleming's microbe for his doctoral thesis. He confirmed Fleming's claims about the instability of the drug and, being unable to extract it himself, made no further investigations. Still, two other American drug companies, Eli Lilly and E. R. Squibb, looked at penicillin's potential. Squibb researchers carried out their own literature search and produced a well-reasoned statement, now a classic in the history of penicillin, concluding that “in view of the slow development, lack of stability and slowness of bacterial action shown by penicillin, its production and marketing as a bactericide
does not appear practicable
.” Penicillin was sidelined in favor of the readily available sulfa drugs.

In 1936, a chemist at Merck was shown a culture of Fleming's
Penicillium notatum
by a physician from New York's Beth Israel Hospital who predicted that more interesting antibiotics were on the way. Three years later, Merck's research director, Randolph Major, asked Dr. Waksman's advice, and he suggested taking penicillin seriously. Other similar agents would probably soon be found, he forecast. Merck immediately hired three new staff members to “study isolation of therapeutic substances from micro-organisms.”

In Britain, the start of the world war had revived interest in Fleming's penicillin. At Oxford University, Howard Florey, an Australian pathologist,
and Ernst Chain, a German Jewish chemist who had fled the Nazis in 1933, began work on purifying penicillin.

In the fall of 1939, Merck returned to Waksman with another proposal. This time the company offered him a second consultancy—of another $150 a month—for information about “chemotherapeutic agents.” “I informed them of my own interest in antibiotics,” Waksman noted later. “They placed another fellowship in my laboratory and engaged me to help Merck in this field of research.” Merck agreed to carry out “chemical, bacteriological and biological tests for the production, purification, plus identification and evaluation and to arrange for clinical trials.” These were the kinds of tests that could not be carried out at Rutgers because of a lack of facilities. In exchange, Merck would have the
exclusive right
to develop any new drug that resulted from the research. The company would pay Rutgers a royalty of 2.5 percent of net sales.

In August 1940, the Oxford team published their first promising results of the use of penicillin on ten patients, and the team was eager to start development. But British industry was overstretched, and under constant air attack. Florey and a colleague, Norman Heatley, brought penicillin to America and found a government not yet at war, and drug companies eager to be the first in the antibiotic market. Merck, Squibb, Lederle Laboratories, and Pfizer & Co., in the East; Abbott, Parke, Davis, and the Up-john Company in the Midwest. Merck agreed to be part of a massive, U.S. government–sponsored war effort to produce penicillin. George Merck sent a telegram to Vannevar Bush, director of the Office of Scientific Research and Development: “
Command me
and my associates ... if you think we can help you.” The Roosevelt administration launched an astonishingly successful example of government-science-industry cooperation, second only in wartime to the atomic bomb project. It would eventually involve ten American and five British firms, combining efforts to make the drug for Allied troops.

George Merck of Merck & Co. with vial. (The Merck Archives, 2011
)

WAKSMAN'S DEAL WITH
Merck caused quite a stir in the offices of the Rutgers administration. They wanted to make sure the university got its share. Like many universities of the day, Rutgers had no policy for dealing with faculty who made patentable discoveries. The most recent case, in 1933, concerned a
professor of pomology
named M. A. Blake, a well-known breeder of peaches who was called the father of the New Jersey peach industry. He was especially proud of his latest nectarine crosses and wanted to apply for a patent.

Whether Blake himself had the right to take out a patent depended on his contract, the Rutgers lawyers advised. If he had been employed specifically to breed nectarines, he would have to assign the patent to Rutgers, but if he was a “general employee” in the fruit-breeding department, and he had bred this spectacular new nectarine in the course of other work, then he would be entitled as an individual to apply for a patent and collect royalties. The lawyers noted, however, in view of Rutgers's duty to make agricultural discoveries available for free to the public, that Professor Blake “might be embarrassed” if he started to profit from the patent. In that case, there was a third way: He could assign it to the nonprofit Rutgers Endowment Foundation, a body originally set up to receive alumni donations. A percentage of the royalties could be paid to the professor, the lawyers said, in line with similar arrangements at other universities.

This quickly became Rutgers's policy; the only question was what percentage, if any, of the royalties to allow the discoverer. In 1937, Rutgers agreed to a
50-50 split
—until it found out that it was being overgenerous compared with other universities, or, as the Rutgers comptroller,
A. S. Johnson, observed, that it had been “decidedly
off on the wrong foot
.” Rutgers reduced the discoverer's share to 25 percent, but even that was above what other institutions were paying; Purdue's was fixed at 20 percent, the University of Wisconsin's at 15 percent. Wisconsin's Alumni Research Foundation director, A. L. Russell, advised Johnson to “keep in mind” that university patents are “to be taken out primarily in the interest of the public rather than for the inventor.”

While the Merck deal with Waksman was being worked out, the first of
Waksman's graduate students
to work on the antibiotic project arrived at the Department of Soil Microbiology. Boyd Woodruff, a tall, confident, genial farmer's son from South Jersey, joined Waksman's laboratory in July 1939. His parents were determined that he should have a university education, but all they could afford was the state-supported agricultural and engineering course at Rutgers. He lived with other students
above the chicken house
, which at that time accommodated 125 white Leghorns. Woodruff earned pocket money selling farm eggs.

Woodruff had found the college experience exciting and sometimes a little overwhelming. He had gone to a concert for the first time, and had celebrated with his fellow students all night in 1937 when the Rutgers football team scored its first victory, 29 to 27, over Princeton since 1889. He graduated in soil chemistry, and Waksman offered him a college fellowship of $900 a year—20 percent more than fellowships elsewhere. The money came from Merck's generous contributions to Waksman, now totaling $3,600 a year.

As a result of his European tours, Waksman attracted students worldwide. Eleven graduate students crammed into the two upstairs laboratories of the Department of Soil Microbiology. They came from China, South America, Europe, and across the United States. At first Woodruff was “terribly discouraged” when Waksman put him to work on composts and gave him little direction. He relied heavily on Waksman's deputy, Robert Starkey, as had most of Waksman's graduate students over the years. As one of them recalled, Starkey was their “
great provider
of materials and receiver of complaints—the equivalent of an assistant in a steel mill. He remembered; he got things done. He told us how to make our cases to Dr. Waksman. Modest to a fault, totally loyal to the Department—it's inconceivable that Dr. Waksman could have operated without him.” Woodruff worked alongside a visiting student from China who was trying to find
out the minimum temperature needed to kill all the harmful bacteria in human feces so that it could be used for compost. Since human feces were not used for compost in America, Waksman suggested that Woodruff should study horse dung, horse urine, and straw, and see how the combination worked. After that, Woodruff moved on to potato scab disease, a serious problem in New Jersey, caused by an actinomycete. The microbe does not grow under acid conditions and is controlled by farmers by adding sulfur to the soils. A bacterium oxidizes the sulfur to sulfuric acid, thus producing the desired soil acidity.

One day, toward the end of 1939, Waksman received word from Merck that the Oxford team was successful in isolating and purifying penicillin. Highly agitated, Waksman appeared in the laboratory and told Woodruff to “
drop everything
. See what these Englishmen have discovered a mold can do. I know the actinomycetes can do better.” Certainly, the Russians had already suggested that the actinomycetes were worth testing.

Waksman took Woodruff to see René Dubos at the Rockefeller Institute so that he could learn the so-called soil-enrichment method that Dubos had used when he discovered tyrothricin. In this method, the researcher adds a disease-producing bacterium to pots of soil over two to three months, hoping this will favor growth of the species of actinomycetes that can kill and feed on that particular bacterium. Woodruff chose
E. coli
as his disease bacterium, adding billions of cells to pots of Rutgers college farm soil he knew was rich in actinomycetes. Each week he counted the surviving
E. coli
by taking a sample of the soil in the pots and growing it on nutrient jellied agar in a petri dish. It was easy to spot the
E. coli
, which appeared as clumps of mold with a distinctive metallic luster. Each time he counted them, the number was reduced until at the end of the three months Woodruff's pots of enriched soil had, as he put it, “become highly efficient
E. coli
killing machines
.” He isolated the sturdy-looking cultures of
Actinomyces antibioticus
, which produced a red chemical substance apparently responsible for the killings. He had found a new antibiotic. Woodruff and Waksman named it actinomycin.

Antibiotics are often difficult to extract and purify, as Fleming had found with penicillin, but actinomycin was relatively easy. Woodruff and Waksman hoped they had struck lucky, but now they needed to make enough actinomycin to study its effect on disease in small animals.

Waksman took a sample to Merck, where the new drug could be produced
on a larger scale than in his laboratory. Then he and Woodruff wrote up their results in a paper that was published in the spring of 1941. Merck's researchers purified and tested actinomycin in animals. The results were shocking. It certainly killed disease bacteria, but it was so toxic it also killed laboratory mice in twenty-four to forty-eight hours. There was no question of testing it on humans. Rat poison was about all it was good for.

Waksman was “
truly excited
” about the discovery, however. “Once I had an actinomycete in my hand which produced antibiotic activity,
everything changed
,” Woodruff recalled. “Waksman started coming into the lab immediately after lunch each day.”

At this early stage, these microbial chemical weapons were so new they had
no specific name
. They were known as “antibiotic substances.” The term came from “antibiosis,” meaning “against life,” a word first used by Louis Pasteur's pupil Jean Paul Vuillemin in 1889 to describe the antagonistic effects of microorganisms. By the 1930s, the use of “antibiotic” as an adjective, as in “antibiotic substance,” was fairly common in the scientific literature, especially among European researchers. Waksman himself used it. But now a proper noun was needed.