Entering a New

Prontosil first came into the French pharmaceutical market on May 8, 1935, in the guise of a copy with the trade name Rubiazol, manufactured by the Roussel Laboratories in Paris. Information on the preparation of Prontosil had been available in France nearly a year earlier. On December 24, 1933, a year after the filing of the German patent application, I.G. Farben had registered a corresponding French patent. This was issued on April 9, 1934, and published on June 21, more than six months before publication of the German patent and almost eight months before Domagk's first publication on Prontosil. Since at this time French patents protecting chemical products did not extend to medicines, Roussel was unobstructed in its efforts to reproduce Prontosil. This accomplished, the compound was turned over to two medical researchers, Constantin Levaditi and Aron Vaisman, at the Institut Alfred Fournier in Paris, for animal trials. Levaditi and Vaisman's paper, published on May 13, 1935, just five days after Rubiazol went on the market, was the first confirmation of Domagk's laboratory results to appear in France. By July, the first reports of clinical trials began to appear, confirming earlier German claims about the effectiveness of Prontosil/Rubiazol in treatment of erysipelas and puerperal fever.1

Roussel Laboratories, Levaditi, and Vaisman were not alone in their rapid response to the news of Prontosil. Within ten days of publication of Domagk's article in February 1935, a copy was in the hands of Ernest Fourneau, director of the Pasteur Institute's section of therapeutic chemistry. Fourneau had already taken note of the French patent eight months earlier, and at that time had written to Edmond Blaise, the scientific director of the chemical and pharmaceutical firm Rhône-Poulenc. Now, with Domagk's paper before him, he acted quickly, speaking to two of his own research staff, Jacques Tréfouël and Daniel Bovet, the same day and soon thereafter conferring with Blaise on a joint course of action. Fourneau and Blaise agreed to pursue a coordinated program of research on compounds related to Prontosil at the Pasteur Institute and at Rhône-Poulenc's laboratories near its factory at Vitry.2

In so doing, Fourneau and Blaise were continuing and extending a commitment to research in chemical therapy and its support by French industry that had its origins in the early years of the century and in which Fourneau had been closely involved. The Pasteur Institute had set up a section of therapeutic chemistry in 1901 as part of an expansion directed by Pasteur's successor, Émile Duclaux. Work on chemotherapy, notably that of Charles Laveran and Félix Mes-nil on trypanosome infections, figured in the institute's Annales even before Paul Ehrlich coined the term in 1906. Fourneau began as a student of pharmacy in Paris, learning organic chemistry from Charles Moureu. A chance meeting with

Emil Fischer, who was staying at his parents' hotel in Biarritz, led Fourneau to a three-year sojourn in Germany, where he continued the study of organic chemistry with Fischer and L. Gatterman in Berlin and with Richard Willstatter in Munich.3

Fourneau returned from Germany convinced that new chemical therapies could best be sought in collaboration with the chemical industry, which was in a position to supply the number and variety of compounds necessary for testing. He entered into an arrangement with the Établissements Poulenc Frères, one of the two parent companies of Rhône-Poulenc, to conduct research with the company's support. Out of this laboratory, which Fourneau headed from 1903 to 1911, came one of the first synthetic local anesthetics, named Stovaine in a pun on the English translation of Fourneau (le fourneau, stove).4

In 1910, the director of the Pasteur Institute, Émile Roux, prompted by Ehrlich's recent breakthrough with Salvarsan and by Fourneau's growing reputation, offered Fourneau the directorship of the institute's section of therapeutic chemistry. Fourneau agreed on the condition that his section maintain the ties he had previously established with Établissements Poulenc Frères. The connection meant both financial support and a continuing flow of intermediates and new compounds. In return, Fourneau agreed with Roux that all his research would be published, contrary to the usual industrial practices of the day. In addition, the company did not attempt to impose research problems, and Fourneau had a free hand to do as he pleased. In practice, this meant not only the search for new synthetic medicines in competition with leading German pharmaceutical research, but also outright copying of existing German products, which were not covered by patents in France, so as to free French pharmaceutical manufacturing from dependence on Germany. The urgency of the latter task increased dramatically with the outbreak of war in 1914.5

Fourneau's service scored early successes, including an oral treatment for syphilis called 190F or Stovarsol, and a sleeping sickness medicine, 270F or Or-sanine. When the Bayer Company kept secret the formula of its sleeping sickness remedy, Bayer 205 or Germanin, Fourneau's team succeeded in synthesizing it independently. Under the name 309F, the compound was distributed to doctors in the French colonies through the overseas Pasteur Institutes and was received enthusiastically.6

The successes of Fourneau's service flowed not only from his leadership and the special combination of steady support and freedom of action built into the arrangement with Établissements Poulenc Frères—what Fourneau proudly called "the mechanism that I created"—but also from the close teamwork he imposed between chemists, medical researchers and clinicians.7 In all of these respects, Fourneau may be compared with Heinrich Horlein, with the notable difference that Horlein acted as executive of the industrial corporation that supported his research enterprise.

In 1928, Établissements Poulenc Frères was absorbed by the Société chimique des Usines de Rhône to form a new and larger entity, Rhône-Poulenc. Both parent companies had their origins in the first half of the nineteenth century, and both had begun to manufacture synthetic organic chemicals in the second half of the century. During or after the World War, Les Usines de Rhône began to manufacture aspirin, and it was their common interest in the aspirin market that brought the two companies together. The leadership of Les Usines de Rhône was composed mainly of men of finance, while that of Les Établissements Poulenc included a majority of pharmacists and doctors of pharmacy. The merger opened new possibilities for pharmaceutical research and development. In addition to its continuing support of Fourneau's service at the Pasteur Institute, where in 1935 Rhône-Poulenc supplied almost half of the budget of what was one of the largest sections of the institute, the company also maintained at the factory at Vitry a staff of about ten chemists and two medical researchers. One of the latter was R.L. Mayer, a German bacteriologist of Alsatian origin, and the other was Bernard Halpern, a Polish doctor who would later occupy the chair of experimental medicine at the Collège de France.8

In February 1935, Fourneau and Blaise initiated a research program that aimed to prepare and test azo compounds that might match or improve upon Pron-tosil's therapeutic action. Chemical and animal experimental work was to proceed simultaneously at Vitry and the Pasteur Institute. They assigned the first stage of the chemical work, the preparation of p-aminobenzenesulfonamide as intermediate, to the Rhône-Poulenc laboratories at Vitry because of its proximity to the factory and greater potential for quantity production. Marcel Delépine, the director of the Vitry laboratories, was a professor at the Collège de France and a long-time friend of both Fourneau and Blaise. A chemist at Vitry delivered the first sample on March 30, and by April 15 had accumulated 200 g. By June, the intermediate was in the hands of chemists P. Goissedet at Vitry and Jacques Tréfouël at the Pasteur Institute.9

Meanwhile, Bovet at the Pasteur Institute strove to replicate Domagk's animal experimental model and to reproduce the results of Domagk's animal trials of Prontosil. To do so, he enlisted the aid of Frédéric Nitti, a bacteriologist working with pathogenic microbes in A. Salimbeni's vaccine service one floor above Bovet's laboratory. Swiss by origin, trained in zoology at Geneva, Bovet had come to Paris in 1929 to work in Rhône-Poulenc's laboratories at Vitry. Instead, a temporary assignment to Fourneau's service turned into a permanent position. Bovet learned to work with laboratory animals and collaborated with Pasteur Institute chemists on a variety of problems, including hypnotics, the chemotherapy of malaria, local anesthetics, and antiseptics. Nitti had interrupted medical studies in his native Italy to join his father in political exile from fascism. Completing his medical studies in Paris with a thesis on use of autovaccines in treatment of bronchial asthma, he entered the Pasteur Institute on a fellowship to continue his research and stayed on in the vaccine service.10

At first, Bovet and Nitti were unable to reproduce Domagk's results. Animal trials carried out with the Pasteur Institute's good collection of bacterial cultures were negative. The cultures were not sufficiently virulent, and Prontosil was without effect. Success came with a fresh culture from a human streptococcal infection, obtained through a friend at the Hôtel-Dieu. The new culture killed mice with the necessary efficiency, and infected mice treated with Prontosil survived. After three months of trials, the collaborators were confident that they had replicated Domagk's animal model and that they could now proceed to screen new compounds.11

By June 1935, conditions were in place for Fourneau's service to proceed with the synthesis and screening of new compounds. The chemical work was done by Jacques and ThereseTrefouel, who had before them the model of Fritz Mietzsch and Joseph Klarer's patent application for Prontosil. By July 13, the Trefouels had produced the first azo dye other than Prontosil, and on July 25 Fourneau wrote to Blaise that he had in hand a compound labeled 1095F that surpassed Prontosil in activity.

Now the pace quickened as the work became routinized. Between July and early November, the Trefouels synthesized forty-four azo derivatives, of which eighteen were sulfonamides. Bovet and Nitti conducted animal trials on some eighty compounds, and Bovet tested each for toxicity after the example of Domagk's February 1935 paper. The urgency of the work was underlined by positive clinical reports on Prontosil and its French homologue, Rubiazol, that arrived in the summer and early fall from both French and German clinicians.12

Prontosil and its copy, Rubiazol, were dyes, and so were many of the compounds synthesized and tested in Fourneau's service between July and November. The Pasteur Institute team saw little reason to depart from a research path that was accessible, clearly marked, and rich in possibilities. That they did so was the result of a discovery that came upon them suddenly and unexpectedly on November 6-8.

The story has been told in detail by Bovet. On the morning of November 6, Bovet and Nitti began with forty mice. Dividing them into groups of four, they gave each an intraperitoneal injection of the same dose of a highly virulent culture of streptococcus. They left one group untreated and gave a second group what they judged to be a curative dose of a Prontosil clone. The remaining eight groups were to be treated with oral doses of new compounds. Only seven such compounds were available, however, leaving an extra group of mice. Bovet recalled that "I thought, why not simply try p-aminophenylsulfonamide, the molecule common to all the compounds being tested?" The large brown glass flask from Vitry containing the intermediate stood before him on the table. He quickly prepared the solution and administered it to the last four mice.

Checking their animals the next day, Bovet and Nitti found the controls dead as expected, while those mice treated with the Prontosil clone were still alive. Only one of the seven new compounds prepared by the Trefouels showed activity, confirming an earlier observation. Arriving finally at the last cage containing the four mice treated with the intermediate, Nitti and Bovet were surprised to find its occupants in good condition. Bovet recalled that his and Nitti's first response was disappointment, since the activity of the intermediate seemed to render all the work to prepare and test dyes of the previous four months "suddenly and completely useless." The let-down was short-lived. "A moment of reflection," Bovet wrote, "was sufficient for us to appreciate the results before us. The last V [indicating survival] was not yet written [on the laboratory record] before Nitti and I were convinced that the future belonged to 'colorless products'. From this moment on the patents of the German chemists would be without value."13

Bovet and Nitti conducted more trials of the intermediate the following week, on larger numbers of animals, with positive results. Once p-aminobenzenesulfonamide's powers had been demonstrated empirically, explanations for them were soon in coming. The Pasteur Institute team noted analogies in the structure of p-aminobenzenesulfonamide (hereafter PABS) and two other chemotherapeutic agents, atoxyl and stibamine. Perhaps the more complex dye molecules of which PABS was a constituent were broken down in the animal organism, releasing the intermediate to act on its own. If so, all of the French and German laboratory and clinical investigators preparing and testing different compounds over the past several months had, unknowingly, merely confirmed the efficacy of a single simpler substance. There was general recognition on Fourneau's service that a real breakthrough had occurred. Jacques Trefouel saw an explanation for what to him had been a puzzling similarity in therapeutic action of very different compounds. Bovet found himself overcome with emotion when he observed even animals infected intravenously recover after treatment with PABS. Fourneau began immediately to plan studies of derivatives belonging to neighboring chemical series.14

Once they had confirmed the action of PABS, its role as the common factor in the activity of a variety of azo dyes seemed obvious to the Pasteur Institute workers, and the experiments that revealed that role were open to anyone. To ensure priority in the discovery, they sought the quickest vehicle of publication. For French researchers in the life sciences, the path of choice was a short note in the

Figure 6.1. Ernest Fourneau (1872-1949) c. 1930 (© Institut Pasteur, reprinted with permission).

Figure 6.2. Daniel Bovet (1907-1992) in his laboratory c. 1955. Swiss by origin, Bovet worked at the Pasteur Institute in Paris from 1924 to 1947. In 1957 he received the Nobel Prize in Physiology or Medicine for his work on antiallergenics and synthetic curares (© Institut Pasteur, reprinted with permission).

Figure 6.3. Jacques (1897-1977) and Thérèse (1892-1978) Tréfouël in their laboratory of therapeutic chemistry at the Pasteur Institute, Paris, c. 1945 (© Institut Pasteur, reprinted with permission).

Figure 6.2. Daniel Bovet (1907-1992) in his laboratory c. 1955. Swiss by origin, Bovet worked at the Pasteur Institute in Paris from 1924 to 1947. In 1957 he received the Nobel Prize in Physiology or Medicine for his work on antiallergenics and synthetic curares (© Institut Pasteur, reprinted with permission).

Figure 6.4. Frédéric Nitti (1905-1947) in his laboratory at the Pasteur Institute, Paris, c. 1945 (© Institut Pasteur, reprinted with permission).

Comptes Rendus des Séances de la Société de Biologie. The Société de biologie, founded in 1848 and counting among its early presidents Pierre Rayer, Claude Bernard, and Paul Bert, continued to thrive in the 1930s in a dual role as testing site for candidates for the Académie de médecine and Académie des sciences and as receptacle for a large scientific correspondence. Since 1929, it had published three volumes of Comptes Rendus a year. In 1935 alone, these volumes comprised 4,756 pages and some 2,000 notes or short communications. Not known for selectivity—no editorial committee reviewed its notes for content, and it was sometimes referred to with affectionate humor as the "Comptes des mille et une nuits" (in a play on Contes des mille et une nuits, Tales of the Thousand and One Nights)—the Comptes Rendus did have the virtue of efficiency, its weekly installments appearing like clockwork. Neither Nitti nor Bovet was a member of the Société, but both knew the secretary and had published other notes in the Comptes Rendus during 1935. The Pasteur Institute team's note was read to the Société on November 23, 1935, and printed on November 28.15

The published note is remarkable in several respects. Although the discovery of the therapeutic efficacy of PABS was made in Fourneau's service, and the collaborators referred to the compound as 1162F in his honor, Fourneau declined to include his name as co-author. Bovet surmised that his chief did so either to advance the careers of his younger colleagues or as a gesture of respect to Horlein and the Elberfeld scientific staff whose prestige might be damaged by the publication. Because notes in the Comptes Rendus were held to strict limits of length, the Pasteur Institute team, facing a week's delay in publication, withheld from the

Figure 6.4. Frédéric Nitti (1905-1947) in his laboratory at the Pasteur Institute, Paris, c. 1945 (© Institut Pasteur, reprinted with permission).

text the protocol of their initial experimental finding. Most striking, in light of Bovet's later narrative of events, is the rational reconstruction that the note imposes on the discovery. The note first reports that the Pasteur Institute team had found that derivatives of Prontosil differing in their physical and chemical properties had an analogous antistreptococcal power. It then continues, in order for such different compounds to act in an almost identical manner, is it not because they undergo in the organism a series of modifications of which the first result will be the cutting of the double bond with formation of aminophenylsulfonamide? This hypothesis led us to study the activity of the hydrochloride of P-aminophenylsulfonamide or 1162F.

In the published note, therefore, the chance conjunction of an extra set of mice and Bovet's casual, impromptu decision to try the intermediate that stood before him on the table is effaced in favor of an experiment proceeding from prior reasoning and a clearly framed hypothesis.16

Assuming that Bovet's later reconstruction was accurate, why did the Pasteur Institute team represent their discovery in 1935 in a way that diverged from the actual sequence of events? A clue may be found in Bovet's remark that once PABS's activity was confirmed, its role as common source of the activity of a variety of azo dyes seemed obvious. So obvious, we may surmise, that retrospective explanation was easily converted into prior hypothesis when the time came to present the discovery to the medical and scientific communities. The Comptes Rendus note of November 1935 appears to be a case not of willful misrepresentation but of the priority frequently ceded to logical order over historical reconstruction in scientific publications.17

The November 1935 note's masking of the element of chance involved in the discovery of PABS's activity, and the representation of the discovery in hypo-theticodeductive terms, may help to account for later speculations such as those of bacteriologist Ronald Hare that attributed a similar discovery to the Elberfeld group. If the reasoning was so clear to the French, so the argument would go, it must have been equally clear to the Germans, who suppressed it only to protect their profits from Prontosil. Bovet's revelation that the reasoning was not obvious to the Pasteur Institute team until after it had made its chance discovery thus undermines the Hare thesis and provides additional reason to suppose that the Elberfeld team had not come to the same conclusions independently.18

Given the importance attached to their discovery by the Pasteur Institute team, and the role that it did eventually have as starting point for a new generation of sulfa drugs, the reception of the discovery in France is surprising and at first sight, baffling. Within the Pasteur Institute itself, response to the discovery outside of Fourneau's own service ranged from indifference to hostility. No other head of service expressed curiosity or desired a part in the follow-up research. What was worse, the finding appears to have created displeasure on the part of the institute's directors. In a letter of March 23, 1936, to director Louis Martin, Fourneau wondered what the institute expected from his laboratory and whether the council clearly perceived the importance of chemotherapy:

It appears that you are very unhappy with what has just happened for an-tistreptococcal products, but I admit that I do not understand at all the reasons for your unhappiness. We can only be very proud to confirm that in a domain where the Germans are past masters, in which they dispose of a considerable number of workers and of raw materials in, so to speak infinite quantities, the French have been able to create for themselves a personal place.19

Two reasons may be suggested for this unfriendly reception. One has to do with the threat posed by PABS, and by bacterial chemotherapy generally, for older biological products on which the Pasteur Institute relied for a part of its financial support. Since the time of Pasteur the institute had derived prestige and income from the development and manufacture of vaccines and serums. By early 1936, Prontosil/Rubiazol had already begun to render antistreptococcal serums obsolete, and the prospect that other serums, for meningococcal and pneumococ-cal infections, dysentery, and plague, would suffer similar fates was clearly discernible. Bovet recalled that he and Jacques Tréfouël had jokingly accused themselves of cutting off the branch on which they sat. It is unlikely that the institute's executives found the idea so amusing. Competition for resources within the institute may have been another ground for the cool response of the other services. Fourneau pushed for an expansion of his unit in light of the increased importance of the chemotherapy of infectious disease. His requests were finally approved by the institute council in November 1938, three years after the PABS discovery, possibly over the resistance of other researchers who resented the already well-supported section of therapeutic chemistry.20

Meanwhile, PABS fared no better in the hands of French manufacturers. Rhône-Poulenc, which might have been expected to move 1162F quickly to clinical trials and to market, instead sought a closely related derivative that could be patented. The result was the synthesis in early 1936 by Rhône-Poulenc chemists of n-benzylsulfonamide. After bacteriological and toxicological tests by company employees Mayer and Halpern and clinical trials by H. Bloch-Michel, M. Conte, and P. Durel, the new compound was marketed in March 1936 by the Société parisienne d'expansion chimique (Spécia) under the trade name Septazine. Bovet and his colleagues were not impressed. Septazine was almost entirely insoluble— a "plaster," in Bovet's and Nitti's terms—and was only moderately effective in Pasteur Institute animal trials; it had no demonstrated clinical advantage over PABS. PABS was kept off the French market until the spring of 1937, well after it had been made available to doctors and patients in Germany and Britain in 1936, and even after its introduction in the United States earlier in 1937. Bovet could only conclude that Rhône-Poulenc and Spécia did not want competition for their specialty in the French market.21

Finally, in March 1937, F. Albert-Buisson, the new managing director of Rhône-Poulenc, visited Fourneau's unit and was persuaded of the importance of PABS. In May 1937, eighteen months after discovery of its therapeutic action at the Pasteur Institute, 1162F (by now called sulfanilamide in the United States) went on the French market under the trade name Septaplix, manufactured by the new pharmaceutical company Théraplix, created by Albert-Buisson and Rhône-Poulenc. It was an immediate success.22

The same month witnessed the opening of the Exposition internationales des arts et des techniques in Paris. The exposition's displays illustrating the material progress of civilization were housed in a new Palais de la découverte (Palace of Discovery) exhibition. Among the booths was one devoted to therapeutic chemistry, in the care of Fourneau, Lecomte de Nouy, and Jacques Tréfouël. Bovet recalled with pride that here "for the first time in history," the most recent results with "synthetic bactericides," the sulfonamides, were presented on large panels alongside the more familiar research on antiparasitic medicines. Jean Perrin, who had conceived the idea for the Palais de la découverte, wrote after its opening that in spite of wars, economic crisis, and other troubles, the development of civilization becomes more and more rapid, thanks to techniques that are ever more supple and effective, in domains more and more extensive. The exposition .. . expresses our legitimate pride in the material progress that we owe to [those techniques]. Almost all have appeared in the last century, developing or applying inventions . . . which seem to have realized or surpassed the desires expressed in our old fairy tales.

Thirty-seven million people attended the exposition, of whom 2,250,000 entered the Palais de la découverte. On the last day, November 25, the organizers awarded a large number of prizes, including a gold medal to Domagk. Bovet recalled that when Perrin spoke of "the domination of matter," and of "the techniques already victorious over smallpox, diphtheria, typhus, and perhaps soon all diseases," he and his colleagues in Fourneau's service almost imagined that Perrin was speaking for them. Perhaps they also shared Perrin's sense that "suddenly, by a sort of glory, we have entered into a new age."23

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