I5OYEA1SAGO paratus. Van Potter had a photoelectric colorimeter I could use in his impressive lab. Harold Rusch, head of the new McArdle Institute, let me use his Soxhlet extractor to measure fat in liver. Professor Homer Adkins, head of the Chemistry Department across the street,was a great help, and so were his faculty who understood diazonium chemistry or physical organic chemistry, which I loved. They had a huge spectrophotometer they let me use in the dead of Vitamin night (2); it wasn’t shielded. The libraries in Chemistry, Physics, and those on the Agriculture Campus had the information that was important to me and that one would not expect to find in our Medical Library. Just now, it would give Karl H. Beyer, Jr. me as much pleasure to write about these and many other friends I made in thisway, whom I still cherish,as about my research. When I received an invitationto write about the abstract I I’m not sure from where the idea of what to do next came. published for the FASEB Meeting in 1942, entitled“The proIt wasn’t very elegant,but I decided to use carbon tetrachlotective action of vitamin C against experimental liver ride to produce damage to the liver (3), perhaps destroying damage,” my first thought was to wish that I had submitted whatever was responsible for getting rid of (deaminating) the an abstract about something else I was doing at the time. amphetamine that was not excreted. (This can be done Something different, but related; something of more obvious without damage to the kidneys.) The dogs that received carbenefit to medicine. bon tetrachloride excreted all (100%) of the dose of amphetaIt does occur to me that the use of the term vitamin C as mine they were given (1). This helped to assure that some bioan essentialnutriment in thiscontext was proper. I used the logical process was metabolizing about half the dose of this chemical equivalent of vitamin C, ascorbic acid, in the title drug. But what could it be? and text of articles that pertained to hydroxylation or oxidaI knew amphetamine (/3-phenylisopropylamine) was not tive deamination of a group of related compounds that deaminated by monoamine oxidase. Blaschko, Richter, and mimicked in some ways the action of the sympathetic Schlossmann (4) had reported that the amine oxidase deami(adrenergic) division of the nervous system. I am not sure nated such compounds (phenylalkylamines) only if the whether this distinctive use of equivalent terms was deliberamino group was on the terminal carbon atom (i.e., comate or intuitive on my part 50 years ago. (I was still a year pounds like13-phenylethylamineand y-phenyl-n-propylamine). or two away from completion of the combined M.D., Ph.D. We confirmed this in the course of related structure/activity programs at Wisconsin.) Much of the time was spent in the studies (5). laboratory. This reasearch with vitamin C derived fortuiBernheim (6) reported that phenylhydrazine would comtously from my work that made use of ascorbic acid as a reapletely inhibit monoamine oxidase in vitro at a concentragent, as follows. tion of 4.6 x 1O M. That order of activity was not impresMy professor, Dr. Walter J. Meek, head of the Departsive, but we tried phenylhydrazine to see whether it would ment of Physiology, introduced me to research by an assignblock the deamination of f3-phenyl-n-propylamine by amine ment to study the effects of a sympathomimetic amine on oxidase in the dog as it did in vitro. It did not block the engastrointestinal motility. That compound, fl-phenylisoprozyme even at hemolytic doses (7). Wells (8) reported that pylamine, is better recognized by all as the amphetamine hydrazine (not phenyihydrazine) produced an almost specific Benzedrine. I enjoyed working with dogs and fellow students central zonal paranchymatous degeneration of the liver in as subjects, and continued to work part of the time with dogs. We found that hydrazine increased excretion of both /3whole animal physiology during my 7 years there and ever and ‘y-phenylpropylamines, like carbon tetrachloride did (7), since. but these experiments gave us no insight into what might be My background in chemistry was the basis for wondering deaminating the phenylisopropylamines. what happened to this orally active compound, the effects of Our lead to the deamination of amphetamine came from which lasted as long. To find out, I needed an analytical the amino acid literature.Euler et al. (9) reported in 1934 method for amphetamine. There was none. With the help of that dehydroascorbic acid was capable of deaminating leua good friend, my former professor of organic chemistry, cine with the formation of aldehydes and ketones. Stotz et al. B. F. Skinner, we worked out a useful analyticalmethod for (10) showed that dehydroascorbic acid could be generated Benzedrine and found that about 43% of a dose was excreted from ascorbic acid in guinea pig liver brei. Keiin and Harin 48 h, whether it was given to dogs orally or subcutanetree (11) reported that the cytochrome oxidase system could ously (1, 2). oxidize ascorbic acid. And so we turned to the ascorbicWhat happened to the other 50% or so? My compulsion dehydroascorbic system with our amines. to know, the lack of biochemically useful equipment available In vitro, the ascorbic-dehydroascorbic acid system deamito me in the Physiology Department, and the concurrence of nated our phenylpropylamines from 30 to 54% of theoretimy broad-minded professor induced me to make friends cal, whether the amino group was on the terminal carbon wherever the equipment I needed could be found. Actually, thiswas a tremendous good fortune: there was no lack of useful equipment to be found here and there on campus. Moreover, I learned a lot of chemistry and biochemistry just Dr. Beyer (APS, ASPET) isVisitingProfessorof Pharmacology, by listening to my friends talk about their research. For inDepartment of Pharmacology, The Milton S. Hershey Medical stance, it was either Al Lehninger or Phil Cohen in PhysioCenter, Pennsylvania State UniversitySchool of Medicine, Herlogical Chemistry who taught me to use their Warburg apshey,PA 17033,USA. This is the twentieth in a series of articles recalling scientific events in FASEB 50 years ago, including reminiscences of the 1942 Annual Meeting.

C and Serendipity,

Fifty

Years

Ago

3316 50 YEARS AGO Vol. 6 November 1992 w.fasebj.org by Iowa State University Serials Acquisitions Dept (129.186.138.35) on January 17, 2019. The FASEB Journal Vol. ${article.issue.getVolume()}, No. ${article.issue.getIssueNum

atom or adjacent thereto (isopropylamines), so long as the compounds contained no hydroxyl group in the ring or on the side chain (12).It was not feasible to try to increase amphetamine excretion by making dogs ascorbic acid-deficient (ascorbutic), so I did the opposite. I established control amphetamine excretion with dogs, then saturated them with ascorbic acid. This reduced amphetamine excretion to 35% or less of the amount given (13). Now I had an answer to my question about amphetamine excretion and inactivation. I should have stopped that line of research right there, but I didn’t. There is always one more experiment thatjust has to be done, or so it seems at the time. In thiscase I wanted to findout what would happen to

amphetamine excretion if we saturated our dogs with ascorbic acid and then gave them the usual dose of carbon tetrachloride. Would the toxin destroy all deamination of the compound under the circumstance of ascorbic acid facilitation? In effect, nothing happened; the toxin did not increase the excretion of am-

phetamine

as it always

had

Karl H. Beyer, Jr. prior to giving vitamin C supplementation. There had to be a reason. We killed and autopsied the dogs as we had done in previous experiments in which the hepatotoxic agents were given. It was evident that vitamin C protected the dogs from liver damage. You could see this protective effect of the vitamin grossly and histomorphically, as well as in the chemistry. If you have read thismuch of my story,you know that the dog experiments just described anticipated the abstract (14) for FASEB of 50 years ago. At this point my research proceeded in two directions.The work that demonstrated in guinea pigs and dogs the protective effectof vitamin C on liver damage and in anesthesiology was published in the clinical literature I have cited (15, 16). On occasion this clinical knowledge can be important and helpful. The ascorbic-dehydroascorbic acid work contributed to the importance of introducing an aliphatic hydroxyl group adjacent to the primary isopropylamine in the compound, metahydroxy-/3-phenylisopropanolarnine. I arrived at, syn-

thesized, and studied this compound at Wisconsin. The laevo form of that compound is stillmarketed by Merck Sharp & Dohme as metaraminol (ARAMINE), used prin-

cipallyto sustain blood pressure in major surgery when itis

needed. Serendipity, such as that which formed the basis for the FASEB abstract in 1942, has added so much of value to the pleasure of my research career these 50 years and more.

REFERENCES 1.Beyer,K. H., and Skinner,J. T. (1940) The detoxication and excretion of phenylisopropylamine (Benzedrine). j PharinacoL Exp. The: 68, 419-432 2.Beyer, K. H. (1942)The color reactionsof sympathomimetic amines with diazonium compounds. j Am. Che,n. Soc. 64, 1318-1322 3. Lainson, P. D., Gardner, G. H., Gustafson, R. K., Marie, E. D., McLean, A. J., and Wells, H. S. (1923) The pharmacology and toxicology of carbon tetrachloride. J. Pharmacol. Exp. Therp. 22, 215-288 4. Blaschko,H., Richter,D.,and Schlossmann, dation of adrenaline and other amines.

H. (1937) The oxiBiochem. j 31,

2187-2196 5. Beyer,

K.

H. (1941)

The

enzymatic

inactivation

of substituted

phenylpropyl-(sympathomimetic)-amines. j PharmacoL Exp. The:. 71, 151-163 6. Bernheim, F. (1940)Note on the actionof copper and phenylhydrazine on certain dehydrogenases.j BioL Chem. 133, 485-489 7.Beyer,K. H., and Lee, W. L. (1942)The fateof certainsympathomimetic amines in the body.j PharmacoL Exp. The: 74, 155-162

8. Wells,

H. 0.

(1908) The

pathological

poisoning. J. Exp. Med 10, 457-465 9. Euler, H. V, Karrer, P., and Zehender,

anatomy F. (1934)

of hydrazine Das

verhalten

von Vitamin C (Ascorbinsaure)und anderer reduktone gegen katheptische und andere enzyme. Helv. Chim. Acta 17, 157-162 10. Stotz,

E., Harrer,

C.J.,

Schultze,

M. 0.,

and King,

C. G. (1938)

The oxidationof ascorbicacid in the presence of guinea pig liver. J. Biol. Chein. 122, 407-418 11.Keilin,D.,and Hartree, E. F.(1938)Cytochrome oxidase.Proc. R. Soc. (Land.) Set. B, 125, 171-186 12.Beyer,K. H. (1942)The ascorbicdehydroascorbicacid system in the synthesis and inactivation of sympathomimetic amines. j Pharinacol. Exp. The: 76, 149-155 13.Beyer,K. H. (1941)The actionofvitamin C and phenol oxidase in the inactivation of (3-phenylisopropylamine (amphetamine). j PhannacoL Exp. The: 71, 394-401 14.Beyer,K. H. (1942)The protectiveactionof vitamin C against experimental liver damage. Federation Proc. 1, 7 (abstr.) (This is the Abstractto which the articlerefers.) 15.Beyer,K. H. (1943)Protectiveactionof vitamin C againstexperimentalhepaticdamage. Arch. Inst. Med. 71, 315-324 16. Beyer, K. H., Stutzman, J. W., and Hafford,B. (1944)The relationofvitamin C to anesthesia. Surg. GynecoL Obstet. 79, 49-56

50 YEARS AGO 3317 Vol. 6 November 1992 w.fasebj.org by Iowa State University Serials Acquisitions Dept (129.186.138.35) on January 17, 2019. The FASEB Journal Vol. ${article.issue.getVolume()}, No. ${article.issue.getIssueNum

Vitamin C and serendipity, fifty years ago.

I5OYEA1SAGO paratus. Van Potter had a photoelectric colorimeter I could use in his impressive lab. Harold Rusch, head of the new McArdle Institute, le...
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