Experiment Eleven

by Peter Pringle

  In those days, Waksman’s students were still not looking for medical applications. “The soil and the microbe,” Waksman wrote, “await the investigator [who] is not looking for practical gains but for explaining the obscure and observing the unknown. The application will doubtless come.” Undoubtedly, Waksman missed a great opportunity. Had he pursued what he had observed with Pasteur’s “prepared mind,” he, not Alexander Fleming, might have been the first to discover an antibiotic.

  But he was not a physician, like Fleming, and Rutgers had no medical department. In his daily life, Waksman was not exposed to faculty discussions about the therapeutic value of “magic bullets” like salvarsan, or the sulfa drugs that followed. In 1932, the German doctor Gerhard Domagk, working at the giant chemical company I. G. Farben, found a bright-red dye that cured mice infected with pathogenic streptococci. This new compound, named Prontosil, was good for fighting a wide range of bacterial infections and later gave rise to the sulfonamides, or sulfa drugs, which had a major impact on the treatment of infectious diseases. (The life of Winston Churchill was saved by a sulfonamide when he developed pneumonia after the Tehran Conference with Stalin and Roosevelt at the end of November 1943.)

  Yet Waksman still lacked funding to expand his research. In America, medical research, like other scientific research, suffered from a lack of public assistance. In the 1920s and ’30s, the National Institutes of Health and the National Science Foundation did not exist. Waksman relied on his wits to attract support. He was a good salesman, a scientist-entrepreneur who never seemed short of industry sponsors.

  He helped tanners find enzymes for defatting hides; brewers, enzymes to clarify beer. He convinced the local mushroom industry that providing funds to investigate a compost mix of alfalfa, peanut shells, and tobacco stalks was a better bet than relying on horse dung from the declining stables of the Philadelphia police department. These links with industry provided rare funding during the depression years, and endeared Waksman’s graduate students to him for providing them with beer and mushroom tastings. There were some unexpected delights. On one famous evening in the college auditorium, female models paraded in then-daring off-the-shoulder evening dresses with fringes of miniature orchids bred in Dr. Waksman’s Department of Soil Microbiology. The event was sponsored by a local businessman hoping to sell the orchids at debutante balls.

  A few independent foundations gave research grants, and in 1932, the National Tuberculosis Association funded Waksman to study the fate of TB germs in people and animals who died of TB and were buried in the soil. He assigned the task to one of his graduate researchers, who found that the tuberculosis bacteria were greatly reduced in some soils. But Waksman did not follow up this interesting result. Similar results were being obtained by other researchers, and Waksman thought they all seemed to lead nowhere. He was not “yet prepared to take advantage of these findings.”

  In late 1935, Fred Beaudette, Rutgers’s director of Poultry Pathology, brought Waksman a test tube containing a TB bacterium specific to poultry that had been destroyed by a fungus that had accidentally contaminated the tube. This was indeed a “happy accident” of the kind that Fleming had encountered seven years earlier with penicillin. Yet Waksman was still not ready to seize the opportunity, this time staring him in the face.

  There was, of course, a perfectly good and understandable reason for not wanting to test a microbe’s ability to destroy pathogenic bacteria: the risk of catching the disease. In his writings, Waksman never mentions this as a factor, but it must have been on his mind. His underfunded laboratories were poorly equipped to protect the workers against infections, or even the hazards of handling dangerous chemicals. In the basement laboratory, protective clothing consisted of worn and torn white lab coats and some “very crusty, black, rubber lab aprons designed to catch splashes of hot acid.” These coats were “hung on spikes driven into the wall” when not in use.

  BUT WAKSMAN COULD not ignore the research coming out of Europe and Russia. In the mid-1930s, the Russians led the world on research into the antagonistic properties of Waksman’s precious actinomycetes. By 1935, Russians had published four papers on the subject; Waksman had published none. A key Russian paper reported that actinomycetes were antagonistic to Bacillus mycoides, one of the standard bacteria tests for antibiotics. The paper concluded, “The question of interrelationships of soil microbes deserves profound research.” Waksman had an enormous advantage over his peers in America in being able to read these papers, not just the English summary that was always included but the whole paper, and some have speculated that the Russian research started to turn his mind toward the possibility of antibiotics, a suggestion he never acknowledged.

  In 1936, at the Second International Congress for Microbiology in London, Alexander Fleming discussed the antibacterial properties of his penicillin, a debate which Waksman later listed as an important event in the evolution of his own thinking. One of Waksman’s graduate students recalled that Fleming’s discussion was when Waksman became “seriously interested” in antibiotic research.

  Later in 1936, Waksman began to study the published papers on warrior microbes and wrote two papers for Soil Science, the journal started at Rutgers by Dr. Lipman. The first paper reviewed the current literature, including the four Russian papers. A measure of Waksman’s absence from basic research in this area is that of the 107 papers he listed, only 2 were written by him.

  In the second paper, also finished in 1936, Waksman and a graduate student, Jackson Foster, tested a fungus, a bacteria, and an actinomycete from a Scottish peat bog. They were all capable of producing “substances which are antagonistic” to other soil microbes when grown in petri dishes containing artificial nutrients. In 1937, another Russian researcher found that antagonistic actinomycetes were “widely distributed” in different soils in the Soviet Union. Of eighty cultures isolated from various soils, forty-seven possessed antagonistic properties, but only twenty-seven were found to be capable of liberating toxic substances into the nutrient agar on a petri dish.

  In 1938, Waksman was especially influenced by the work of one of his former students, René Dubos. A Frenchman who had qualified in agriculture and immigrated to America in 1924 after hearing Lipman speak at the conference on soil science in Rome, Dubos worked for his Ph.D. under Waksman at Rutgers. He discovered a soil microbe that produced an enzyme capable of breaking down cellulose, the key ingredient of plant stalks and tree bark, and turning it into plant food. Similar work was being carried out at the Rockefeller Institute for Medical Research, in New York City, where Dubos later moved. There, he eventually isolated a bacterial enzyme that destroyed the sugary coat of the bacteria that causes pneumonia. Unfortunately, the enzyme was too toxic to be used by humans suffering from pneumonia, but Dubos was sufficiently encouraged to begin the first systematic search for antibiotics in the soil.

  In 1939, he found an antibacterial agent produced by a bacterium, Bacillus brevis, and named it tyrothricin. The Rockefeller biochemist Rollin Hotchkiss helped him recognize that it was made up of two compounds, tyrocidin and gramicidin. Tyrocidin was toxic to mice, but gramicidin cured experimental infections in mice, without side effects. Gramicidin was too toxic to be administered to humans intravenously, but it was effective when used on open wounds. The Russians produced their own version of Dubos’s discovery, known as gramicidin S (for Soviet), and used it throughout World War Two as their main antibiotic.

  That same year, the Russians struck again. Two researchers, N. A. Krassilnikov and A. I. Korenyako, again found that many species of actinomycetes produced antibiotics. The Russians concluded that “one cannot escape the possibility of using the bacterial factor of actinomycetes” to treat bacterial diseases. For the first time, they discovered two that were active, ever so slightly, against Mycobacteria, the group that causes tuberculosis. To anyone searching for a cure for TB, it was a powerful clue that such an antibiotic might be found.

  By late 1939—in the wake of pioneering rese
arch by the Russians, a major discovery by Dubos in New York, and the beginning of the war in Europe—Waksman, the soil microbiologist who had pledged his life to microbes that could be used in plants and industry, was finally ready to change the direction of his research to look for antibiotics to cure human diseases. All he needed was a sponsor.

  4 • The Sponsor

  Twelve miles down the railroad track from the Rutgers campus is Rahway, New Jersey, once an old Indian settlement and a stop on the stagecoach run from New York to Philadelphia. Since 1903, Rahway has been the home of Merck & Co., then and now one of the most important pharmaceutical concerns in the country. Friedrich Jacob Merck opened the original family-owned apothecary, the Engel-Apotheke (Angel Pharmacy), in 1656 in Darmstadt, Germany. In 1827, the Merck company started producing morphine, codeine, and cocaine. By the 1890s, Merck was selling so many products in America that the family dispatched the twenty-four-year-old George Merck to set up shop there. He settled in Manhattan, bought 150 acres of Rahway, and later sent his son, George Jr., a blond, blue-eyed giant at six feet five inches, to Harvard. On George Sr.’s death in 1926, his son took over the business.

  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 payi
ng; 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.