- Email: [email protected]
Role of Hydrazine in Carcinogenesis
ADVANCES I N CANCER RESEARCH, VOL. 30
ROLE OF HYDRAZINE IN CARCINOGENESIS Joseph Ba16 Department of Pathological Anatomy, Semmelweis Medical University, Budapest. Hungary
I. Introduction
..........................................................
............ 11. Toxic Effect of Hydrazine ............................. ........................ 111. Hydrazine-Induced Alteration in Rat Liver
........... IV. Occurrence of Hydrazines in the Environm V. The Oncogenicity of Isonicotinylhydrazide ............................... VI. Experiments of H. Druckrey in the Production of Tumors with Hydrazine Compounds ............................... VII. Production o VIII. Hydrazine-Caused Cancer .............................................. IX. Does IN H Produce Tumors in Humans? ................................. X. Methylhydrazine Derivatives, a New Class of Cytotoxic Agents . . . . . . . . . . . . XI. Hydrazine Therapy in Hodgkin's Disease ................................ XII. Summary ............................................................. References ...........................................................
1.51
1.52 152
1.53 1.53 15.5 158 1.58 1.59 160 161 161
I62
I. Introduction
Hydrazine NH,-NH,, as a diamide, is produced when sodium hypochlorite acts in the presence of excess ammonia:
+ NaOCl = NH,CI + NaOH NH&I + NH, = NH,-NH,*HCl
NH,
The hydrochloride of hydrazine, NH,-NH,.HCl is a colorless, oily liquid, which fumes in air, is slightly more viscous than water, and has a penetrating ammoniacal odor. It boils at 113.5"C, freezes at 2"C, and can be stored for years if sealed in glass and kept in a cool, dark place. It is miscible with water, methanol, and ethanol, but is insoluble in ether, chloroform, and benzene. It is caustic to skin and mucous membranes, and is carcinogenic. It produces hydrazine hydrate with water and hydrazine salts with acids. Hydrazine reacts with aldehydes and ketones, forming hydrazones. Metal hydrazides are combinations of metals with hydrazine. Its alkylated derivative, dimethylhydrazine, together with hydrogen peroxide, nitrogen oxide, and saltpeter is used as rocket fuel. Prior to 1875 only hydrazo compounds, derivatives of hydrazine, were known. In that same year Emil Fischer suggested the name hydrazine. 1.51
Copyright @ 1979 by Academic Preps. Inc. All rights of reproduction in any form reserved ISBN n-12-m63oo
I52
JOSEPH B A L 6
Isonicotinylhydrazide or isoniazid (INH) is used in the treatment of tuberculosis. Maleic hydrazide is employed to control suckering of tobacco. Hydrazine has military application in rocket propellants, hydrazine insecticides are common environmental pollutants, and pharmacological effects of hydrazine compounds are being exploited in a group of antidepressive agents which have monoamine oxidase-inhibiting effects (Toth, 1976). II. Toxic Effect of Hydrazine
Borissow (18941, Poduschka (l900), and Pohl (1902) established that hydrazine exerts toxic effects in experimental animals. In 1908 Underhill and Kleiner reported that hydrazine produced alterations in the liver of bitches. Following injection of doses of 0.1 gm, restlessness was observed, with augmentation of the heart beat, followed by respiratory difficulty and general paresis. Coincident with these symptoms, variable quantities of protein, bile pigment, and allantoin crystals were observed in the urine. Wells (1908) found that hydrazine seems to be almost specific to the cytoplasm of the liver parenchyma. However, surprisingly, hydrazine was not at all toxic for the myocardium, kidney, and red blood cells. After hydrazine intoxication, recovery is ,very rapid causing no anatomical and histological changes, unlike that found with other hepatic toxins. In experimental intoxication there is accumulation of fat in the periportal and midzonal regions, sparing the pericentral region of the liver cells, 24 hours after injection, with loss of glycogen. In fasting animals neither lipid nor glycogen can be found. Parenteral feeding showed that the liver can store glycogen, although its quantity after 24 hours was only 40% of the controls. Gideon Wells, professor of Pathology at the University of Chicago in 1908, published a paper on the pathohistology of hydrazine in the J . Exp. M c ~ d .10, 457. He also poisoned and then examined the tissues of a few other dogs, as well as of cats and guinea pigs. Ill. Hydrazine-Induced Alteration in Rat Liver
Much of the current knowledge of hydrazine toxicity is based on studies of the rat. Amenta and Johnston (1962)found that animals became relatively quiet within 2 hours after injection. Four hours after adminis-
ROLE OF HYDRAZINE IN CARCINOGENESIS
153
tration of hydrazine the liver was pale, and upon microscopic examination droplets of fat could be found in the periportal and midzonal regions. The lipid concentration reached maximal value at 24 hours and then rapidly decreased. Lewis and Izume (1926) showed that parenteral glucose could not be converted into liver glycogen in hydrazine-treated rabbits. Amenta and Johnston (1962) emphasized that hydrazine sulfate intoxication does not produce either liver necrosis or inflammation. IV. Occurrence of Hydrazines in the Environment
In nature hydrazine occurs in tobacco and mushrooms. In tobacco, hydrazine is present in the leaves and in the smoke. Liu and Hoffmann (1973) and Liu et al. (1974) found that tobacco smoke from a single cigarette contained 30 ng of hydrazine. Burning of amino acids or protein also liberates hydrazine. Levenberg (1964) found that the common edible mushroom, Agaricrrs bisporirs found in the United States contains a hydrazine named agaritin. The y-glutamyltransferase dissociates agaritin into L-glutamate and 4hydroxymethylphenylhydrazine. In Europe an indigenous mushroom, Agaricirs liortensis, produces a substance similar to agaritin. A wild mushroom, Gyrornitra escirlenta, contains methylhydrazine. If this mushroom is cooked in an open pot, methylhydrazine evaporates. However, if the lid is placed on the pot the methylhydrazine cannot vaporize, with the consequent effect that the mushroom becomes poisonous (List and Luft, 1968, 1969). Gowing and Leeper (1955) observed that pineapple seedlings sprinkled with ethylhydrazine flowered earlier than usual. Hydrazines find application not only in industry and horticulture but also in medical therapy. INH, though routinely used in the therapy of tuberculosis, is not used for prevention because of its tumorigenicity. Among the diseases of blood-forming organs, in the therapy of polycytemia Vera the use of phenylhydrazine has been introduced. Math6 e t al. (1963) applied methylhydrazine in the therapy of Hodgkin's disease. V. The Oncogenicity of lsonicotinylhydrazide
Domagk ef a!. (1952) in 1951 synthesized the drug Neoteben [isonicotinylhydrazide or isoniazid (INH)], a most effective remedy for tuberculosis. Balo (1959, 1965) and Balo et al. (1961) noted that following the intraperitoneal administration of Neoteben, tumors of the lung arise in
I54
JOSEPH B A L 6
mice. The authors employed INH produced by a pharmaceutical firm in Budapest. In a footnote from the paper by Juhhsz el al. (1957), Hackmann and Hecht, representing the Bayer Company, stated that Neoteben was nontumorigenic. Following a request from the author, however, Domagk provided us with Neoteben from the Bayer Company; it was proved that Bayer INH was equally tumorigenic. The establishment of the carcinogenicity of INH caused surprise in the scientific community, but was confirmed in several laboratories. Mori and Yasuno (1959) produced lung tumors in dd strain mice by oral administration of INH. Mori el al. (1960) found that in five groups of 15-20 mice each, 50%- 100% of mice treated with INH developed lung tumors, whereas in the two control groups, lung tumors were found in only 13%. The authors held that the carbamyl group was responsible for tumorigenicity. Matsumoto et al. (1960) also observed tumors after the oral administration of INH. Wolfart (1960) at the Robert Koch Clinic in Freiburg concluded on the basis of the data at that time that the tumorigenicity of INH could not be denied. On the other hand, Viallier and Casanova (1960) were not in agreement. Schwan (1961, 1962), on the contrary, supported the fact that INH produced tumors. Wagner and Moritz (1962) found that in animal experiments or in tissue culture application of small doses of INH promoted tumor growth, whereas large doses had inhibitory effects. In 1962 Weinstein and Kinoshita using BALB and C5, black mice obtained mainly lung tumors. Biancifiori et al. (1964) experimented with the CBA/Cb/Se strain of mice with the same result. Beer and Schaffner (1959) treated a 65-year-old woman for hypertension. In her last six weeks she was given 600 mg of P-phenylhydrazine and died with symptoms of hepatitis. Engbaek et al. (1965) compared respective dosages and concluded that the tumorigenic doses used in animal experiments did not surpass those commonly used in the treatment of human beings. According to Pompe (1956) lupus vulgaris seldom leads to carcinoma (only 0.5%) but in lupus treated with INH, carcinoma occurred in 4.6%. Biancifiori and Ribacchi (1962) and Biancifiori et al. (1964) endeavored to ascertain which part of the INH compound was carcinogenic. Using mice of the BALB/c strain, they administered sodium isonicotinate to one group, to another, isonicotinic acid hydrazine, and to a third group, hydrazine sulfate. In animals given isonicotinic acid hydrazide or hydrazine sulfate the development of lung tumors occurred. In animals that received sodium isonicotinate few tumors developed. On the basis of these findings it is concluded that the hydrazine moiety of isonicotinic
ROLE OF HYDRAZINE IN CARCINOGENESIS
155
acid hydrazine is primarily responsible for the turnongenic action. According to JuhAsz et al. (1967) hydrazine hydrate dissolved in physiological saline and all compounds from which hydrazine is liberated were carcinogenic. Seven and Biancifiori (1968) also emphasized the carcinogenic effect of hydrazine. Presently there are no data to confirm or deny that the administration of INH is a human cancer hazard. In Haddow’s department, Roe et al. (1965) already stressed that it is necessary to make long-term, repeated INH trials to learn whether INH therapy predisposes to lung cancer or to cancer of other organs. VI. Experiments of H. Druckrey in the Production of Tumors with Hydrazine Compounds
Nitrosamines and their derivatives are generally carcinogenic, with a wide spectrum. The first observations in this field were made by Barnes and Magee (1954) who showed that N-dimethylnitrosamines were potent poison to liver in both man and experimental animals. Moreover, oral administration to rats has resulted in liver adenomas or carcinomas (Magee and Barnes, 1956). Since nitrosamines are very simple aliphatic compounds, with many chemical variations, Druckrey et al. (1961) have used them as models for studies of the relationship between chemical structure and carcinogenicity. They scrutinized systematically the homologous series of dialkylnitrosamines and acyl alkylnitrosamides (Druckrey et af. 1964, 1966, 1967a), as well as many derivatives of N-nitroso compounds, among them the dimethyl- and diethylhydrazine. Among the more than 65 compounds examined, they found that nearly all symmetrically substituted dialkylnitrosamines have produced carcinomas of the liver (Schmahl et a l . 1960). Nonsymmetrical dialkylnitrosamines selectively induced tumors of the esophagus (Druckrey et al. 1964). After subcutaneous injection of cyclic nitrosamines, tumors of the nasal cavity (ethmoturbinalia) have been encountered. Methylnitrosourea was effective in selectively inducing brain tumors in rats (Druckrey and Preussmann, 1964), and treatment with N-methyl, N-nitrosourea has resulted in malignant growth of the brain and the spinal cord (Druckrey and Preussmann, 1965; Thomas, 1965; Thomas and Kersting, 1964). The selective carcinogenic effects of different nitroso compounds provoking tumors in specific organs provided Druckrey et al. (1967a) with the impetus to construct the so-called “diazoalkane theory.” According to this theory, the different alkyl compounds of nitrosamines were transformed to the respective azoalkanes. This proceeded in different organs
156
JOSEPH B A L 6
by the action of organ-specific hydroxylases forming the respective azoxyalkanes. This important step would actually determine organotropy. The proposed biochemical mechanism was supported by the observed carcinogenic effects of azoethane and azoxyethane (Druckrey et a l . , 1965). Assuming that all aliphatic azoderivatives would be dealkylated in higher organisms through oxidation, producing diazoalkanes, the carcinogenic effect of dialkylhydrazine seemed to be evident. The following mechanism was proposed in the degradation of alkylhydrazines: H n
R-CH,-N
H-N H
Qli
4
symmetrical dialkylhydrazine
-R'
I\
lH
specific hydroxylase
1/ d H
a a
R-CH,-NH-NH--C
-R'
I
h ydroxydialkylhydrazine
\ oxidation
I
R--CH,-NAN-C
\
-R' OH
-H,O
diazoalkane
R-CH,-N=N-CH,-R'
J
oxidation R-CH,-N
\
N-N+-CH,--RR'
hydroxydiazoalkane
reduction
= N-CH,-R'
i
diazoxyalkane
/
heterolysis
+ C+H,-RR'
+ N,
end products
From the completed first step onward all following reactions through the alkylating diazoalkane occur spontaneously. The initial a-hydroxylation is considered, thus, the clue for the observed remarkable organotropic effects. In view of the use of 1,2-dialkylhydrazines as rocket fuels, Druckrey et a l . (1965) started their comprehensive experimental work on hydrazines with 1,2-diethyIhydrazine and 1,2-dimethyIhydrazine. Druckrey et
ROLE OF HYDRAZINE IN CARCINOGENESIS
157
al. (1966) performed the following experiment. Forty-five rats of the BD strain, in 3 groups, obtained weekly 1,2-diethylhydrazine, given subcutaneously in dosages of 100, 50, and 25 mg/kg body weight, respectively, in neutralized aqueous solution. After 126, 191, and 296 days, 43 of the total of 45, animals developed tumors. There were 8 reticulosarcomas, 5 stem cell and blast cell leukemias, 15 esthesioneuromas, ethmoturbinalias, 6 gliomas or gliomas with mesodermal mixed tumors of the brain, 2 adventitial sarcomas of the brain, 1 malignant neuronoma of the cauda equina, 1 subcutaneous sarcoma, and 15 adenocarcinomas of the breast out of 20 female rats. The latter corresponded closely to human breast cancer and could be transplanted as reticulosis and leukemia to animals of the same strain. These results showed remarkable similarities with the carcinogenic effects of azoethane. All the above tumors, except for one did not occur at the subcutaneous site of injection but in remote organs. From this fact it was clear that the carcinogenic effect of diethylhydrazine is exerted not directly but is due rather to a transformed product which arises during its metabolic activation. Druckrey et af. (1967) carried out further experiments with 1,2-dirnethylhydrazine using rats of BD strain, injecting 21 mg/kg, in water (pH 6.5) subcutaneously. Following a mean induction period of 184 ? 30 days, all animals died with multiple tumors of the intestine, as follows: duodenum, 6; small intestine, 5; colon, 10; rectum, 8. The histological diagnosis was adenocarcinoma, which infiltrated the whole thickness of the aforementioned organs. In these experiments, Druckrey et af. (1967) selectively induced intestinal cancer in rats using dimethylhydrazine. It was concluded that neither 1,2-diethylhydrazine nor 1,2-dimethylhydrazine have direct carcinogenic effects, but require metabolic activation through organ-specific hydroxylases as do the nitroso compounds. Finally, a conversion by oxidative dealkylation to diazoalkanes occurs. The organotropy of the hydrazines was further confirmed by circumstantial evidence presented by Druckrey et af. (1967a). They discovered that the symmetrical 1 ,2-dimethylhydrazine (CH,-NH-NH-CH, 1 was one hundred times more carcinogenic, producing tumors selectively in the colon, than the nonsymmetrical analog, 1 , I-N-dimethylhydrazine
158
JOSEPH B A L 6
VII. Production of Polyps and Tumors in the Intestinal Canal
Introduction of dimethylbenzanthracene or benzpyrene into the intestine rectally does not cause polyps or tumors in the bowel. Many years ago pathologists investigated the genesis of papillomas and adenomata of the bowel. Dukes (1947) studied the origin of rectal adenomata and papillomas. Cole and McKalen (1963) attempted to produce polyps of the large intestine experimentally. Bertalanffy ( 1960) examined the occurrence of mitoses in the epithelium of the alimentary canal of rats, and Bertalanffy and Nagy (1958) observed mitoses in human duodenum. In rats, Druckrey et al. (1966) brought about the development of intestinal cancer with 1,2-dimethylhydrazine. In similar investigations by Wiebecke et a l . (1969) with mice and rats subcutaneous injections of dimethylhydrazine produced adenomatous polyps and adenocarcinoma in the intestinal canal. With weekly subcutaneous hydrazine administration, Thurnherr et u1. (1973) produced hyperplasia, polyps, and adenoma of the epithelium of mucous membrane. CFI strain mice received weekly 20 mg/kg of 1,2dimethylhydrazine. In the early stage, focal hyperplasia was observed in the crypts of glands. Troncale et ul. (1971) and Lipkin et a / . (1963) observed the regeneration of rectal epithelial cells. Recently, Tan et al. (1976) reviewed methods of selective in t i f r o cultivation of adenocarcinomas. Thus, subcutaneous administration of hydrazine derivatives produces polyps and tumors of the intestinal canal. VIII. Hydrazine-Caused Cancer
Cancer caused by hydrazine differs considerably from other cancers in that it does not develop at the site of administration but is absorbed from the circulation, and its carcinogenic effect is exerted in the organs where its metabolism occurs. Targets frequently are the cells of intestinal mucosa, lung, or parenchymal organs. On occasion, tumors arise simultaneously with or following another tumor either in the same or other organs. Schmahl et al. (1976) administered 30 mg/kg I ,2-dimethylhydrazine (DMH) to newborn Sprague-Dawley rats monthly beginning from the second day after birth through 10 months. After 330 days, 72% of the animals developed adenocarcinomas of the large intestine, 17% developed squamous cell carcinomas of the external auditory canal, 13% had adenocarcinomas of the kidney, and I I % had liver carcinomas. Simultaneous administration of immune-depressant or enzyme-activating
ROLE OF HYDRAZINE IN CARCINOGENESIS
159
agents did not influence the development of these different types of cancer. However, on a vegetarian diet the occurrence of liver and kidney tumors diminished significantly and cancers of the intestine and ear duct increased. Druckrey et al. (1966, 1967) stated that DMH selectively produces intestinal cancer. Schmahl et al. (1976) did not agree, however. Their difference of opinion may be explained by the fact that the experiments of Druckrey et al. (1966, 1967) were carried out on 3-month-old animals, whereas Schmahl et al. (1976) used newborn rats. The application of different hydrazine compounds, the age of animals, as well as their state of nutrition, and probably other factors influence differences in the target organ. Druckrey (1966, 1967) and his co-workers found in a comparison of I ,2-diethylhydrazine and I ,2-dimethylhydrazine that this small difference in the structure of the hydrazines brings about considerable disparity in the sites and structures of the tumors. Toth and Wilson (1971) succeeded in producing tumors of the blood vessels in Swiss mice. If they applied symmetrical dimethylhydrazine within 7 days after birth, blood vessel tumors occurred in 95% of the females and 92% of the males, whereas occurrence in control females was only 3% and in males 1%. The blood vessel tumors were mostly angiosarcomas localized in the muscles, perirenal fat, liver, and pararenal, paradidymal tissue. On the other hand, nonsymmetrical dimethylhydrazines yielded mainly lung tumors. The investigation of Reddy et al. (1974) on germ-free rodents indicated that the intestinal microflora plays a modifying role in colon carcinogenesis not only by liberating an active metabolite but also by supplying promoters or accelerators to act on colon mucosa.
IX. Does INH Produce Tumors in Humans? At present it is not yet decided whether the administration of INH to patients with pulmonary tuberculosis increases their risk of developing cancer. Every tuberculous patient who dies is by no means examined at necropsy and no systemic comparison has been made of the incidence of cancer in tuberculous patients treated with INH. Peacock and Peacock (1966) concluded that INH is an acceptable risk when used as a curative drug for tuberculosis, but should not be used prophylactically in healthy infants. It should be regarded as a potential carcinogen, and it would be prudent to assume there is some degree of risk involved in its use. I t
160
JOSEPH B A L 6
should be used only for therapy and, in any case, should be limited, as far as possible, in the duration of its application. X. Methylhydrazine Derivatives, a New Class of Cytotoxic Agents
Zeller rt ul. (1963) when testing a series of hydrazines for another purpose found that I-methyl-2-benzylhydrazine had a pronounced tumorinhibiting effect. Screening of several hundred compounds revealed some forty to be efficient tumor inhibitors, among which I-methyl-2-p-(isopropylcarbamoy1)benzylhydrazine hydrochloride and I-methyl-2-allophanoylhydrobromide were chosen for extended biological and clinical trials. The tumor-inhibiting effects of this new class of cytostatic methyl derivatives are as follows. The growth of the Ehrlich carcinoma in solid ascitic form, the Crocker-sarcoma 256, and the epithelioma of the uterus T 8 are definitely inhibited. I-Methyl-2-p-(isopropylcarbamoyl)benzylhydrazine hydrochloride (I) and 1-methyl-2-p-allophanoylbenzylhydrazine hydrobromide (11) distinguish themselves especially through their strong cytostatic activity. CH, ONH-CH-NCI-HCI CHa
CHa-N H-N H--CH, I
The cytostatically effective methylhydrazine derivative Natulan is described in a series of chemical, experimental, and clinical papers. The metabolism of this tumor-inhibiting methylhydrazine derivative proceeds according to a similar pattern in man, dog, and rat. Initially, a very rapid oxidation of the hydrazine group occurs with the formation of an azo compound, then cleavage and further oxidative degradation follows. The major portion of the drug is excreted in the urine as N-isopropylterephthalamic acid. Rutishauser and Bollag (1967) in their earlier experiments found that methylbenzylhydrazine (MBH) hinders mitosis: later they proposed that this procarbazine impedes the synthesis of DNA.
ROLE OF HYDRAZINE IN CARCINOGENESIS
161
XI. Hydrazine Therapy in Hodgkin’s Disease
According to Math6 et al. (1963) l-methyl-2-p-(isopropylcarbamoyl) benzylhydrazine chlorhydrate, a radiomimetic agent, has proved of considerable value in the treatment of Hodgkin’s disease. This drug was administered to 22 patients and resulted in 7 apparently complete remissions and 8 incomplete remissions, partial failure in 5 patients, and complete failure of disease which had been treated before and was resistant to radiotherapy, alkylating agents, or vinblastine and also in patients in early stages of the disease which have not been treated previously. Some incomplete remissions were obtained in cases of reticulosarcoma and lymphosarcoma. XII. Summary
The author and his pupils in 1957, and the author later, in 1962, in Perugia (Ba16, 1965)established that isoniazid (INH), a compound used in the treatment of tuberculosis, stimulates the growth of tumors. Juhasz et al. (1966)confirmed this in 1967 and subsequent studies found that INH produces tumors in many species of animals. Roe ef al. (1965) stressed the necessity of ascertaining whether INH therapy may also induce tumors in human beings. On the other hand, many researchers are currently working to find, among the great variety of hydrazine compounds, some that possess therapeutic effects against cancer, including a study on ethylhydrazine being carried out by the Hoffmann La Roche Co. Cancer caused by hydrazine differs from other cancers, such as those caused by tar, which develop at the site of application. Cancer due to hydrazine does not develop at the site of application but rather in the organs in which metabolism occurs. Frequent targets are intestinal mucosa, lungs, parenchymal organs, or even the nervous system. Following development of cancer in one organ, tumors may occur in different organs Weitzel ef al. (1963).Oswald and Kriiger (1969)point out that cancer induction by hydrazines injected into the subcutaneous tissue is influenced by its absorption and modification by local tissues, by age, and by the nutritional state. Toth and Wilson (1971) observed that both symmetry and nonsymmetry of hydrazines influence the development of hydrazine tumors. The hydrazine tumor cannot be attributed to the effect of hydrazine alone or its derivatives, but it is the result of the mutual effects of hydrazine and its products of metabolism. From a variety of studies the conclusion is inescapable that all hydrazines are carcinogenic with the
162
JOSEPH BALO
possible exception of methylhydrazine (Natulan) (Roe r t ul., 1967; Toth, 1975; d’Allesandri et al., 1963). However the studies of Gold (1966, 1973) who observed that hydrazine sulfate and various hydrazides inhibit growth of Walker 256 intravascular carcinoma, B- 16 melanoma, Murphy-Sturm lymphosarcoma, and L- I2 10 solid leukemia, suggest that hydrazines may find a place in tumor therapy (Issekutz, 1968). REFERENCES Amenta, J. S., and Johnston, E. H. (1962). Lab. Inivst. 11, 956-962. Balo, J . (1959). “Berliner Symposium uber Fragen der Carcinogenese.” Akad. Verlag, Berlin. Ba16, J. (1962). Miinch. M i d . Wschr. 104, 1424-1428. Balo, J. (1965). Qrrrrdr. Cot$ Cancer 3, 623-636. Univ. Perugia, Italy. Balo, J., Juhasz, J., and Kendrey, G. (1961). Z . Kri~bsfbrsch.59, 561-567. Barnes, J . M . , and Magee, P. N. (1954). Br. J . fndrrstr. Med. 11, 167. Beer, D. T.. and Schaffner, F. (1959). J . A m . Med. Assoc. 171, 887-889. Bertalanffy, F. D. (1960). Acta Anat. 40, 130- 148. Bertalanffy, F. D., and Nagy, K. (1958). Anat. Rec. 130, 271-272. Biancifiori, C., and Ribacchi, R. (1962). Narwe (London) 194, 488-489. Biancifiori, C.. Bucciarelli, E., Clayson, D. B., and Santilli, F. E. (1964). Br. J . Cancer 18, 543-550. Borissow, P. (1894). Z. Pliysiol. Chrm. 19, 499-510. Cole, J . W., and McKalen, A. (1963). Cancer 16, 998-1002. d’Alessandri, A., Keel, H. J., Bollag, W., and Martz. G. (1963). Sc/iw. Mrd. Wschr. 93, 1018-1024. Domagk. G.. Offe, R. A., and Siefken. W. (1952). Dtscli. Med. Wschr. 77, 573-578. Druckrey, H., and Preussmann, R . ( I 964). Nntrrrioissi.nsc/iqfifn 51, 144. Druckrey, H . , and Preussmann, R. (1965). Z. KrebsJi,rsch. 66, 389. Druckrey. H., Ivankovic, S . , Mennel, H. D., and Preussmann. R. (1964). Z. Krrbsforscli. 66, 138- 150. Druckrey, H., Preussmann, R.,Schmahl, D., and Muller, N . (I961 ). Naturii,issensc/iurffPn 48, 134-135. Druckrey, H., Preussmann, R., Matzkies, F., and Ivankovic, S. (1966). NatrrrM~issi.tisr./iafL ten 53, 557-558. Druckrey, H., Preussmann, R., Matzkies, F.. and Ivankovic, S. ( I 967). Natririi,issc,nsL./ia/~ I P N 54, 285-286. Druckrey. H . . Preussmann, R.. Ivankovic, S., and Schmahl, D. (l967a). Z. Kreh.~fhrsc/i. 69, 103-201. Druckrey, H., Preussmann, R., Ivankovic, S., Schmidr, C. H., So, B. T., and Thomas, C. (1965). Z . Krebsforsch. 67, 31-45. Dukes, C. E. (1947). Proc. R . Soc. Med. 40, 829-830. Engbaeck, H. C., Bentzon, M. W.,Heegaard. H., and Christensen, 0. (1965). Acta Path. Microbiol. S c a d . 65, 69-83. Gold, J. (1966). Cancer Res. 26, 695-705. Gold, J . (1973). Oncologv 27, 69-80.
ROLE O F HYDRAZINE IN CARCINOGENESIS
163
Gowing, D. P., and Leeper, R. W. (1955). Science 122, 1267. Issekutr, B., Sr. (1968). Onco/ogy 22, 173-184. Juhasz, J., Ba16, J., and Kendrey, G. (1957). Z. Krehsforsch. 62, 188-197. JuhBsz, J., Balo, J., and Szende, B. (1966). Nciture (Loridon) 210, 1377. Juhasz, J., Balo, J., and Szende, B. (1967). Z. Krehsforsch. 70, 150-156. Levenberg, B. (1964). J. B i d . Chein. 239, 2267-2273. Lewis, H. B., and Izume, S. (1926). J . B i d . Chern. 71, 33. Lipkin, M., Bell, B., and Sherlock, P. (1963). J . Clin. Invest. 42, 767-776. List, P. H., and Luft, P. (1968). Arch. Phurm. 301, 294-308. List, P. H., and Luft, P. (1969). Arch. Pharin. 302, 143-146. Liu, Y.Y., and Hoffmann, D. (1973). A n d . Chem. 45, 2270. Liu, Y. Y., Schmeltz, I., and Hoffmann, D. (1974). Anal. Chein. 46, 885-889. Magee, P. N., and Barnes, J. M. (1956). Br. J . Cancer 10, 114. Matht, D., Schweisguth, O., Schneider, M., Amiel, J . L., Berumen, Z., Brule, G., Cattan, A., and Schwarzenberg, L. (1963). Lnncet 2, 1077-1080. Matsumoto, K., Mori, K., and Jasuno, A. (1960). Gann 51, 91. Mori, K., and Jasuno, A. (1959). Gann 50, 107- 110. Mori, K., Jasuno, A., and Matsumoto, K. (1960). Gan/i 51, 83-89. Oswald, H., and Kriiger, F. W. (1969). Arzneimittel Forsch. 19, 1891-1892. Peacock, A., and Peacock, P. R. (1966). Br. J . Cancer 20, 307-325. Poduschka, R. (1900). Arch. Exp. Puthol. Phunn. 44, 59-67. Pohl, J. (1902). Arch. Exp. Pathol. Pharm. 48, 367-375. Pompe, K. (1956). Derin. Wschr. 133, 105-108. Reddy, B. S . , Weisberger, J . N., Narisawa, T., and Wynder, E. E. (1974). Cancer R r s . 34, 2368-2372. Roe, F. T . C., Boyland, E., and Haddow, A. (1965). Br. Med. J . 1, 1550. Roe, F. T. C., Grant, G. A., and Milican, D. M. (1967). Ncrfure (Lo,idon) 216, 375-376. Rutishauser, A., and Bollag, W. (1967). Experientiu 23, 222-223. Schmahl, D., Preussmann, R., and Hamperl, H. (1960). Naturwis.senschrrfte/i 47, 89- 196. Schmahl, D., Danisman, A., Habs, M., and Diehl, B. (1976). Z. Krehsforsch. Klin. Onkol. 86, 89-94. Schwan, S. (1961). Prrthol. Pol. 12, 53. Schwan, S. (1962). Pfithol. Pol. 13, 185. Severi, L., and Biancifiori, C. (1968). J . N u t / . Caricer Inst. 41, 331-340. Tan, M. H., Holyoke, E. D., and Goldrosen, M. H. (1976). J. Nor/. Cancer Inst. 56, 871873. Thomas, C. (1965). Z. Krehsforsch. 67, 1-30. Thomas, C., and Kersting, G. (1964). Nuturwissenschajien 51, 144- 145. Thurnherr, N., Deschner, E. E., Stonehill, E. H., and Lipkin, M. (1973). Cuncer Res. 33, 940- 945. Toth, B. (1975). Cancer Res. 35, 3693-3697. Toth, B. (1976). Cancer R e s . 36, 917-921. Toth, B., and Wilson, R. B. (1971). A m . J. Puth. 64, 585-600. Troncale, F., Hertz, R., and Lipkin, M. (1971). Cancer Res. 31, 463-467. Underhill, F., and Kleiner, I. S. (1908). J . B i d . Chern. 4, 165. Viallier, J., and Casanova, F. (1960). C . R . S . B i d . (Paris) 154, 985-987. Wagner, H., and Moritz, R. (1962). Arch. Geschwulstforsch. 19, 123- 129. Weinstein, H. J., and Kinosita, R. (1962). J. Lab. Clirz. Med. 60, 1025. Weitzel, G., Schneider, F., Fretzdorf, A. M., Seynsche, K., and Finger, H. (1963). Z. Physiol. Chern. 334, 1-25.
164
JOSEPH BAL6
Wells, H. G . (1908). J . B i d . Chem. 4, 165. Wells, H . G . (1908). J . E.rp. Med. 10, 457-464. Wiebecke, B., Lohrs, V., Girnmy, .I. and , Eder, M. (1969). Z. Ges. Exp. Meci. 149, 277278. Wolfart, W. (1960). Dtsch. Mecl. Wschr. 85, 1655- 1657. Zeller, P., Gutmann. H . , Hegedus, B., Kaiser, A . , Langemann, A., and Muller, M. (1963). Experientiri 19, 129.
ROLE OF HYDRAZINE IN CARCINOGENESIS Joseph Ba16 Department of Pathological Anatomy, Semmelweis Medical University, Budapest. Hungary
I. Introduction
..........................................................
............ 11. Toxic Effect of Hydrazine ............................. ........................ 111. Hydrazine-Induced Alteration in Rat Liver
........... IV. Occurrence of Hydrazines in the Environm V. The Oncogenicity of Isonicotinylhydrazide ............................... VI. Experiments of H. Druckrey in the Production of Tumors with Hydrazine Compounds ............................... VII. Production o VIII. Hydrazine-Caused Cancer .............................................. IX. Does IN H Produce Tumors in Humans? ................................. X. Methylhydrazine Derivatives, a New Class of Cytotoxic Agents . . . . . . . . . . . . XI. Hydrazine Therapy in Hodgkin's Disease ................................ XII. Summary ............................................................. References ...........................................................
1.51
1.52 152
1.53 1.53 15.5 158 1.58 1.59 160 161 161
I62
I. Introduction
Hydrazine NH,-NH,, as a diamide, is produced when sodium hypochlorite acts in the presence of excess ammonia:
+ NaOCl = NH,CI + NaOH NH&I + NH, = NH,-NH,*HCl
NH,
The hydrochloride of hydrazine, NH,-NH,.HCl is a colorless, oily liquid, which fumes in air, is slightly more viscous than water, and has a penetrating ammoniacal odor. It boils at 113.5"C, freezes at 2"C, and can be stored for years if sealed in glass and kept in a cool, dark place. It is miscible with water, methanol, and ethanol, but is insoluble in ether, chloroform, and benzene. It is caustic to skin and mucous membranes, and is carcinogenic. It produces hydrazine hydrate with water and hydrazine salts with acids. Hydrazine reacts with aldehydes and ketones, forming hydrazones. Metal hydrazides are combinations of metals with hydrazine. Its alkylated derivative, dimethylhydrazine, together with hydrogen peroxide, nitrogen oxide, and saltpeter is used as rocket fuel. Prior to 1875 only hydrazo compounds, derivatives of hydrazine, were known. In that same year Emil Fischer suggested the name hydrazine. 1.51
Copyright @ 1979 by Academic Preps. Inc. All rights of reproduction in any form reserved ISBN n-12-m63oo
I52
JOSEPH B A L 6
Isonicotinylhydrazide or isoniazid (INH) is used in the treatment of tuberculosis. Maleic hydrazide is employed to control suckering of tobacco. Hydrazine has military application in rocket propellants, hydrazine insecticides are common environmental pollutants, and pharmacological effects of hydrazine compounds are being exploited in a group of antidepressive agents which have monoamine oxidase-inhibiting effects (Toth, 1976). II. Toxic Effect of Hydrazine
Borissow (18941, Poduschka (l900), and Pohl (1902) established that hydrazine exerts toxic effects in experimental animals. In 1908 Underhill and Kleiner reported that hydrazine produced alterations in the liver of bitches. Following injection of doses of 0.1 gm, restlessness was observed, with augmentation of the heart beat, followed by respiratory difficulty and general paresis. Coincident with these symptoms, variable quantities of protein, bile pigment, and allantoin crystals were observed in the urine. Wells (1908) found that hydrazine seems to be almost specific to the cytoplasm of the liver parenchyma. However, surprisingly, hydrazine was not at all toxic for the myocardium, kidney, and red blood cells. After hydrazine intoxication, recovery is ,very rapid causing no anatomical and histological changes, unlike that found with other hepatic toxins. In experimental intoxication there is accumulation of fat in the periportal and midzonal regions, sparing the pericentral region of the liver cells, 24 hours after injection, with loss of glycogen. In fasting animals neither lipid nor glycogen can be found. Parenteral feeding showed that the liver can store glycogen, although its quantity after 24 hours was only 40% of the controls. Gideon Wells, professor of Pathology at the University of Chicago in 1908, published a paper on the pathohistology of hydrazine in the J . Exp. M c ~ d .10, 457. He also poisoned and then examined the tissues of a few other dogs, as well as of cats and guinea pigs. Ill. Hydrazine-Induced Alteration in Rat Liver
Much of the current knowledge of hydrazine toxicity is based on studies of the rat. Amenta and Johnston (1962)found that animals became relatively quiet within 2 hours after injection. Four hours after adminis-
ROLE OF HYDRAZINE IN CARCINOGENESIS
153
tration of hydrazine the liver was pale, and upon microscopic examination droplets of fat could be found in the periportal and midzonal regions. The lipid concentration reached maximal value at 24 hours and then rapidly decreased. Lewis and Izume (1926) showed that parenteral glucose could not be converted into liver glycogen in hydrazine-treated rabbits. Amenta and Johnston (1962) emphasized that hydrazine sulfate intoxication does not produce either liver necrosis or inflammation. IV. Occurrence of Hydrazines in the Environment
In nature hydrazine occurs in tobacco and mushrooms. In tobacco, hydrazine is present in the leaves and in the smoke. Liu and Hoffmann (1973) and Liu et al. (1974) found that tobacco smoke from a single cigarette contained 30 ng of hydrazine. Burning of amino acids or protein also liberates hydrazine. Levenberg (1964) found that the common edible mushroom, Agaricrrs bisporirs found in the United States contains a hydrazine named agaritin. The y-glutamyltransferase dissociates agaritin into L-glutamate and 4hydroxymethylphenylhydrazine. In Europe an indigenous mushroom, Agaricirs liortensis, produces a substance similar to agaritin. A wild mushroom, Gyrornitra escirlenta, contains methylhydrazine. If this mushroom is cooked in an open pot, methylhydrazine evaporates. However, if the lid is placed on the pot the methylhydrazine cannot vaporize, with the consequent effect that the mushroom becomes poisonous (List and Luft, 1968, 1969). Gowing and Leeper (1955) observed that pineapple seedlings sprinkled with ethylhydrazine flowered earlier than usual. Hydrazines find application not only in industry and horticulture but also in medical therapy. INH, though routinely used in the therapy of tuberculosis, is not used for prevention because of its tumorigenicity. Among the diseases of blood-forming organs, in the therapy of polycytemia Vera the use of phenylhydrazine has been introduced. Math6 e t al. (1963) applied methylhydrazine in the therapy of Hodgkin's disease. V. The Oncogenicity of lsonicotinylhydrazide
Domagk ef a!. (1952) in 1951 synthesized the drug Neoteben [isonicotinylhydrazide or isoniazid (INH)], a most effective remedy for tuberculosis. Balo (1959, 1965) and Balo et al. (1961) noted that following the intraperitoneal administration of Neoteben, tumors of the lung arise in
I54
JOSEPH B A L 6
mice. The authors employed INH produced by a pharmaceutical firm in Budapest. In a footnote from the paper by Juhhsz el al. (1957), Hackmann and Hecht, representing the Bayer Company, stated that Neoteben was nontumorigenic. Following a request from the author, however, Domagk provided us with Neoteben from the Bayer Company; it was proved that Bayer INH was equally tumorigenic. The establishment of the carcinogenicity of INH caused surprise in the scientific community, but was confirmed in several laboratories. Mori and Yasuno (1959) produced lung tumors in dd strain mice by oral administration of INH. Mori el al. (1960) found that in five groups of 15-20 mice each, 50%- 100% of mice treated with INH developed lung tumors, whereas in the two control groups, lung tumors were found in only 13%. The authors held that the carbamyl group was responsible for tumorigenicity. Matsumoto et al. (1960) also observed tumors after the oral administration of INH. Wolfart (1960) at the Robert Koch Clinic in Freiburg concluded on the basis of the data at that time that the tumorigenicity of INH could not be denied. On the other hand, Viallier and Casanova (1960) were not in agreement. Schwan (1961, 1962), on the contrary, supported the fact that INH produced tumors. Wagner and Moritz (1962) found that in animal experiments or in tissue culture application of small doses of INH promoted tumor growth, whereas large doses had inhibitory effects. In 1962 Weinstein and Kinoshita using BALB and C5, black mice obtained mainly lung tumors. Biancifiori et al. (1964) experimented with the CBA/Cb/Se strain of mice with the same result. Beer and Schaffner (1959) treated a 65-year-old woman for hypertension. In her last six weeks she was given 600 mg of P-phenylhydrazine and died with symptoms of hepatitis. Engbaek et al. (1965) compared respective dosages and concluded that the tumorigenic doses used in animal experiments did not surpass those commonly used in the treatment of human beings. According to Pompe (1956) lupus vulgaris seldom leads to carcinoma (only 0.5%) but in lupus treated with INH, carcinoma occurred in 4.6%. Biancifiori and Ribacchi (1962) and Biancifiori et al. (1964) endeavored to ascertain which part of the INH compound was carcinogenic. Using mice of the BALB/c strain, they administered sodium isonicotinate to one group, to another, isonicotinic acid hydrazine, and to a third group, hydrazine sulfate. In animals given isonicotinic acid hydrazide or hydrazine sulfate the development of lung tumors occurred. In animals that received sodium isonicotinate few tumors developed. On the basis of these findings it is concluded that the hydrazine moiety of isonicotinic
ROLE OF HYDRAZINE IN CARCINOGENESIS
155
acid hydrazine is primarily responsible for the turnongenic action. According to JuhAsz et al. (1967) hydrazine hydrate dissolved in physiological saline and all compounds from which hydrazine is liberated were carcinogenic. Seven and Biancifiori (1968) also emphasized the carcinogenic effect of hydrazine. Presently there are no data to confirm or deny that the administration of INH is a human cancer hazard. In Haddow’s department, Roe et al. (1965) already stressed that it is necessary to make long-term, repeated INH trials to learn whether INH therapy predisposes to lung cancer or to cancer of other organs. VI. Experiments of H. Druckrey in the Production of Tumors with Hydrazine Compounds
Nitrosamines and their derivatives are generally carcinogenic, with a wide spectrum. The first observations in this field were made by Barnes and Magee (1954) who showed that N-dimethylnitrosamines were potent poison to liver in both man and experimental animals. Moreover, oral administration to rats has resulted in liver adenomas or carcinomas (Magee and Barnes, 1956). Since nitrosamines are very simple aliphatic compounds, with many chemical variations, Druckrey et al. (1961) have used them as models for studies of the relationship between chemical structure and carcinogenicity. They scrutinized systematically the homologous series of dialkylnitrosamines and acyl alkylnitrosamides (Druckrey et af. 1964, 1966, 1967a), as well as many derivatives of N-nitroso compounds, among them the dimethyl- and diethylhydrazine. Among the more than 65 compounds examined, they found that nearly all symmetrically substituted dialkylnitrosamines have produced carcinomas of the liver (Schmahl et a l . 1960). Nonsymmetrical dialkylnitrosamines selectively induced tumors of the esophagus (Druckrey et al. 1964). After subcutaneous injection of cyclic nitrosamines, tumors of the nasal cavity (ethmoturbinalia) have been encountered. Methylnitrosourea was effective in selectively inducing brain tumors in rats (Druckrey and Preussmann, 1964), and treatment with N-methyl, N-nitrosourea has resulted in malignant growth of the brain and the spinal cord (Druckrey and Preussmann, 1965; Thomas, 1965; Thomas and Kersting, 1964). The selective carcinogenic effects of different nitroso compounds provoking tumors in specific organs provided Druckrey et al. (1967a) with the impetus to construct the so-called “diazoalkane theory.” According to this theory, the different alkyl compounds of nitrosamines were transformed to the respective azoalkanes. This proceeded in different organs
156
JOSEPH B A L 6
by the action of organ-specific hydroxylases forming the respective azoxyalkanes. This important step would actually determine organotropy. The proposed biochemical mechanism was supported by the observed carcinogenic effects of azoethane and azoxyethane (Druckrey et a l . , 1965). Assuming that all aliphatic azoderivatives would be dealkylated in higher organisms through oxidation, producing diazoalkanes, the carcinogenic effect of dialkylhydrazine seemed to be evident. The following mechanism was proposed in the degradation of alkylhydrazines: H n
R-CH,-N
H-N H
Qli
4
symmetrical dialkylhydrazine
-R'
I\
lH
specific hydroxylase
1/ d H
a a
R-CH,-NH-NH--C
-R'
I
h ydroxydialkylhydrazine
\ oxidation
I
R--CH,-NAN-C
\
-R' OH
-H,O
diazoalkane
R-CH,-N=N-CH,-R'
J
oxidation R-CH,-N
\
N-N+-CH,--RR'
hydroxydiazoalkane
reduction
= N-CH,-R'
i
diazoxyalkane
/
heterolysis
+ C+H,-RR'
+ N,
end products
From the completed first step onward all following reactions through the alkylating diazoalkane occur spontaneously. The initial a-hydroxylation is considered, thus, the clue for the observed remarkable organotropic effects. In view of the use of 1,2-dialkylhydrazines as rocket fuels, Druckrey et a l . (1965) started their comprehensive experimental work on hydrazines with 1,2-diethyIhydrazine and 1,2-dimethyIhydrazine. Druckrey et
ROLE OF HYDRAZINE IN CARCINOGENESIS
157
al. (1966) performed the following experiment. Forty-five rats of the BD strain, in 3 groups, obtained weekly 1,2-diethylhydrazine, given subcutaneously in dosages of 100, 50, and 25 mg/kg body weight, respectively, in neutralized aqueous solution. After 126, 191, and 296 days, 43 of the total of 45, animals developed tumors. There were 8 reticulosarcomas, 5 stem cell and blast cell leukemias, 15 esthesioneuromas, ethmoturbinalias, 6 gliomas or gliomas with mesodermal mixed tumors of the brain, 2 adventitial sarcomas of the brain, 1 malignant neuronoma of the cauda equina, 1 subcutaneous sarcoma, and 15 adenocarcinomas of the breast out of 20 female rats. The latter corresponded closely to human breast cancer and could be transplanted as reticulosis and leukemia to animals of the same strain. These results showed remarkable similarities with the carcinogenic effects of azoethane. All the above tumors, except for one did not occur at the subcutaneous site of injection but in remote organs. From this fact it was clear that the carcinogenic effect of diethylhydrazine is exerted not directly but is due rather to a transformed product which arises during its metabolic activation. Druckrey et af. (1967) carried out further experiments with 1,2-dirnethylhydrazine using rats of BD strain, injecting 21 mg/kg, in water (pH 6.5) subcutaneously. Following a mean induction period of 184 ? 30 days, all animals died with multiple tumors of the intestine, as follows: duodenum, 6; small intestine, 5; colon, 10; rectum, 8. The histological diagnosis was adenocarcinoma, which infiltrated the whole thickness of the aforementioned organs. In these experiments, Druckrey et af. (1967) selectively induced intestinal cancer in rats using dimethylhydrazine. It was concluded that neither 1,2-diethylhydrazine nor 1,2-dimethylhydrazine have direct carcinogenic effects, but require metabolic activation through organ-specific hydroxylases as do the nitroso compounds. Finally, a conversion by oxidative dealkylation to diazoalkanes occurs. The organotropy of the hydrazines was further confirmed by circumstantial evidence presented by Druckrey et af. (1967a). They discovered that the symmetrical 1 ,2-dimethylhydrazine (CH,-NH-NH-CH, 1 was one hundred times more carcinogenic, producing tumors selectively in the colon, than the nonsymmetrical analog, 1 , I-N-dimethylhydrazine
158
JOSEPH B A L 6
VII. Production of Polyps and Tumors in the Intestinal Canal
Introduction of dimethylbenzanthracene or benzpyrene into the intestine rectally does not cause polyps or tumors in the bowel. Many years ago pathologists investigated the genesis of papillomas and adenomata of the bowel. Dukes (1947) studied the origin of rectal adenomata and papillomas. Cole and McKalen (1963) attempted to produce polyps of the large intestine experimentally. Bertalanffy ( 1960) examined the occurrence of mitoses in the epithelium of the alimentary canal of rats, and Bertalanffy and Nagy (1958) observed mitoses in human duodenum. In rats, Druckrey et al. (1966) brought about the development of intestinal cancer with 1,2-dimethylhydrazine. In similar investigations by Wiebecke et a l . (1969) with mice and rats subcutaneous injections of dimethylhydrazine produced adenomatous polyps and adenocarcinoma in the intestinal canal. With weekly subcutaneous hydrazine administration, Thurnherr et u1. (1973) produced hyperplasia, polyps, and adenoma of the epithelium of mucous membrane. CFI strain mice received weekly 20 mg/kg of 1,2dimethylhydrazine. In the early stage, focal hyperplasia was observed in the crypts of glands. Troncale et ul. (1971) and Lipkin et a / . (1963) observed the regeneration of rectal epithelial cells. Recently, Tan et al. (1976) reviewed methods of selective in t i f r o cultivation of adenocarcinomas. Thus, subcutaneous administration of hydrazine derivatives produces polyps and tumors of the intestinal canal. VIII. Hydrazine-Caused Cancer
Cancer caused by hydrazine differs considerably from other cancers in that it does not develop at the site of administration but is absorbed from the circulation, and its carcinogenic effect is exerted in the organs where its metabolism occurs. Targets frequently are the cells of intestinal mucosa, lung, or parenchymal organs. On occasion, tumors arise simultaneously with or following another tumor either in the same or other organs. Schmahl et al. (1976) administered 30 mg/kg I ,2-dimethylhydrazine (DMH) to newborn Sprague-Dawley rats monthly beginning from the second day after birth through 10 months. After 330 days, 72% of the animals developed adenocarcinomas of the large intestine, 17% developed squamous cell carcinomas of the external auditory canal, 13% had adenocarcinomas of the kidney, and I I % had liver carcinomas. Simultaneous administration of immune-depressant or enzyme-activating
ROLE OF HYDRAZINE IN CARCINOGENESIS
159
agents did not influence the development of these different types of cancer. However, on a vegetarian diet the occurrence of liver and kidney tumors diminished significantly and cancers of the intestine and ear duct increased. Druckrey et al. (1966, 1967) stated that DMH selectively produces intestinal cancer. Schmahl et al. (1976) did not agree, however. Their difference of opinion may be explained by the fact that the experiments of Druckrey et al. (1966, 1967) were carried out on 3-month-old animals, whereas Schmahl et al. (1976) used newborn rats. The application of different hydrazine compounds, the age of animals, as well as their state of nutrition, and probably other factors influence differences in the target organ. Druckrey (1966, 1967) and his co-workers found in a comparison of I ,2-diethylhydrazine and I ,2-dimethylhydrazine that this small difference in the structure of the hydrazines brings about considerable disparity in the sites and structures of the tumors. Toth and Wilson (1971) succeeded in producing tumors of the blood vessels in Swiss mice. If they applied symmetrical dimethylhydrazine within 7 days after birth, blood vessel tumors occurred in 95% of the females and 92% of the males, whereas occurrence in control females was only 3% and in males 1%. The blood vessel tumors were mostly angiosarcomas localized in the muscles, perirenal fat, liver, and pararenal, paradidymal tissue. On the other hand, nonsymmetrical dimethylhydrazines yielded mainly lung tumors. The investigation of Reddy et al. (1974) on germ-free rodents indicated that the intestinal microflora plays a modifying role in colon carcinogenesis not only by liberating an active metabolite but also by supplying promoters or accelerators to act on colon mucosa.
IX. Does INH Produce Tumors in Humans? At present it is not yet decided whether the administration of INH to patients with pulmonary tuberculosis increases their risk of developing cancer. Every tuberculous patient who dies is by no means examined at necropsy and no systemic comparison has been made of the incidence of cancer in tuberculous patients treated with INH. Peacock and Peacock (1966) concluded that INH is an acceptable risk when used as a curative drug for tuberculosis, but should not be used prophylactically in healthy infants. It should be regarded as a potential carcinogen, and it would be prudent to assume there is some degree of risk involved in its use. I t
160
JOSEPH B A L 6
should be used only for therapy and, in any case, should be limited, as far as possible, in the duration of its application. X. Methylhydrazine Derivatives, a New Class of Cytotoxic Agents
Zeller rt ul. (1963) when testing a series of hydrazines for another purpose found that I-methyl-2-benzylhydrazine had a pronounced tumorinhibiting effect. Screening of several hundred compounds revealed some forty to be efficient tumor inhibitors, among which I-methyl-2-p-(isopropylcarbamoy1)benzylhydrazine hydrochloride and I-methyl-2-allophanoylhydrobromide were chosen for extended biological and clinical trials. The tumor-inhibiting effects of this new class of cytostatic methyl derivatives are as follows. The growth of the Ehrlich carcinoma in solid ascitic form, the Crocker-sarcoma 256, and the epithelioma of the uterus T 8 are definitely inhibited. I-Methyl-2-p-(isopropylcarbamoyl)benzylhydrazine hydrochloride (I) and 1-methyl-2-p-allophanoylbenzylhydrazine hydrobromide (11) distinguish themselves especially through their strong cytostatic activity. CH, ONH-CH-NCI-HCI CHa
CHa-N H-N H--CH, I
The cytostatically effective methylhydrazine derivative Natulan is described in a series of chemical, experimental, and clinical papers. The metabolism of this tumor-inhibiting methylhydrazine derivative proceeds according to a similar pattern in man, dog, and rat. Initially, a very rapid oxidation of the hydrazine group occurs with the formation of an azo compound, then cleavage and further oxidative degradation follows. The major portion of the drug is excreted in the urine as N-isopropylterephthalamic acid. Rutishauser and Bollag (1967) in their earlier experiments found that methylbenzylhydrazine (MBH) hinders mitosis: later they proposed that this procarbazine impedes the synthesis of DNA.
ROLE OF HYDRAZINE IN CARCINOGENESIS
161
XI. Hydrazine Therapy in Hodgkin’s Disease
According to Math6 et al. (1963) l-methyl-2-p-(isopropylcarbamoyl) benzylhydrazine chlorhydrate, a radiomimetic agent, has proved of considerable value in the treatment of Hodgkin’s disease. This drug was administered to 22 patients and resulted in 7 apparently complete remissions and 8 incomplete remissions, partial failure in 5 patients, and complete failure of disease which had been treated before and was resistant to radiotherapy, alkylating agents, or vinblastine and also in patients in early stages of the disease which have not been treated previously. Some incomplete remissions were obtained in cases of reticulosarcoma and lymphosarcoma. XII. Summary
The author and his pupils in 1957, and the author later, in 1962, in Perugia (Ba16, 1965)established that isoniazid (INH), a compound used in the treatment of tuberculosis, stimulates the growth of tumors. Juhasz et al. (1966)confirmed this in 1967 and subsequent studies found that INH produces tumors in many species of animals. Roe ef al. (1965) stressed the necessity of ascertaining whether INH therapy may also induce tumors in human beings. On the other hand, many researchers are currently working to find, among the great variety of hydrazine compounds, some that possess therapeutic effects against cancer, including a study on ethylhydrazine being carried out by the Hoffmann La Roche Co. Cancer caused by hydrazine differs from other cancers, such as those caused by tar, which develop at the site of application. Cancer due to hydrazine does not develop at the site of application but rather in the organs in which metabolism occurs. Frequent targets are intestinal mucosa, lungs, parenchymal organs, or even the nervous system. Following development of cancer in one organ, tumors may occur in different organs Weitzel ef al. (1963).Oswald and Kriiger (1969)point out that cancer induction by hydrazines injected into the subcutaneous tissue is influenced by its absorption and modification by local tissues, by age, and by the nutritional state. Toth and Wilson (1971) observed that both symmetry and nonsymmetry of hydrazines influence the development of hydrazine tumors. The hydrazine tumor cannot be attributed to the effect of hydrazine alone or its derivatives, but it is the result of the mutual effects of hydrazine and its products of metabolism. From a variety of studies the conclusion is inescapable that all hydrazines are carcinogenic with the
162
JOSEPH BALO
possible exception of methylhydrazine (Natulan) (Roe r t ul., 1967; Toth, 1975; d’Allesandri et al., 1963). However the studies of Gold (1966, 1973) who observed that hydrazine sulfate and various hydrazides inhibit growth of Walker 256 intravascular carcinoma, B- 16 melanoma, Murphy-Sturm lymphosarcoma, and L- I2 10 solid leukemia, suggest that hydrazines may find a place in tumor therapy (Issekutz, 1968). REFERENCES Amenta, J. S., and Johnston, E. H. (1962). Lab. Inivst. 11, 956-962. Balo, J . (1959). “Berliner Symposium uber Fragen der Carcinogenese.” Akad. Verlag, Berlin. Ba16, J. (1962). Miinch. M i d . Wschr. 104, 1424-1428. Balo, J. (1965). Qrrrrdr. Cot$ Cancer 3, 623-636. Univ. Perugia, Italy. Balo, J., Juhasz, J., and Kendrey, G. (1961). Z . Kri~bsfbrsch.59, 561-567. Barnes, J . M . , and Magee, P. N. (1954). Br. J . fndrrstr. Med. 11, 167. Beer, D. T.. and Schaffner, F. (1959). J . A m . Med. Assoc. 171, 887-889. Bertalanffy, F. D. (1960). Acta Anat. 40, 130- 148. Bertalanffy, F. D., and Nagy, K. (1958). Anat. Rec. 130, 271-272. Biancifiori, C., and Ribacchi, R. (1962). Narwe (London) 194, 488-489. Biancifiori, C.. Bucciarelli, E., Clayson, D. B., and Santilli, F. E. (1964). Br. J . Cancer 18, 543-550. Borissow, P. (1894). Z. Pliysiol. Chrm. 19, 499-510. Cole, J . W., and McKalen, A. (1963). Cancer 16, 998-1002. d’Alessandri, A., Keel, H. J., Bollag, W., and Martz. G. (1963). Sc/iw. Mrd. Wschr. 93, 1018-1024. Domagk. G.. Offe, R. A., and Siefken. W. (1952). Dtscli. Med. Wschr. 77, 573-578. Druckrey, H., and Preussmann, R . ( I 964). Nntrrrioissi.nsc/iqfifn 51, 144. Druckrey, H . , and Preussmann, R. (1965). Z. KrebsJi,rsch. 66, 389. Druckrey. H., Ivankovic, S . , Mennel, H. D., and Preussmann. R. (1964). Z. Krrbsforscli. 66, 138- 150. Druckrey, H., Preussmann, R.,Schmahl, D., and Muller, N . (I961 ). Naturii,issensc/iurffPn 48, 134-135. Druckrey, H., Preussmann, R., Matzkies, F., and Ivankovic, S. (1966). NatrrrM~issi.tisr./iafL ten 53, 557-558. Druckrey, H., Preussmann, R., Matzkies, F.. and Ivankovic, S. ( I 967). Natririi,issc,nsL./ia/~ I P N 54, 285-286. Druckrey. H . . Preussmann, R.. Ivankovic, S., and Schmahl, D. (l967a). Z. Kreh.~fhrsc/i. 69, 103-201. Druckrey, H., Preussmann, R., Ivankovic, S., Schmidr, C. H., So, B. T., and Thomas, C. (1965). Z . Krebsforsch. 67, 31-45. Dukes, C. E. (1947). Proc. R . Soc. Med. 40, 829-830. Engbaeck, H. C., Bentzon, M. W.,Heegaard. H., and Christensen, 0. (1965). Acta Path. Microbiol. S c a d . 65, 69-83. Gold, J. (1966). Cancer Res. 26, 695-705. Gold, J . (1973). Oncologv 27, 69-80.
ROLE O F HYDRAZINE IN CARCINOGENESIS
163
Gowing, D. P., and Leeper, R. W. (1955). Science 122, 1267. Issekutr, B., Sr. (1968). Onco/ogy 22, 173-184. Juhasz, J., Ba16, J., and Kendrey, G. (1957). Z. Krehsforsch. 62, 188-197. JuhBsz, J., Balo, J., and Szende, B. (1966). Nciture (Loridon) 210, 1377. Juhasz, J., Balo, J., and Szende, B. (1967). Z. Krehsforsch. 70, 150-156. Levenberg, B. (1964). J. B i d . Chein. 239, 2267-2273. Lewis, H. B., and Izume, S. (1926). J . B i d . Chern. 71, 33. Lipkin, M., Bell, B., and Sherlock, P. (1963). J . Clin. Invest. 42, 767-776. List, P. H., and Luft, P. (1968). Arch. Phurm. 301, 294-308. List, P. H., and Luft, P. (1969). Arch. Pharin. 302, 143-146. Liu, Y.Y., and Hoffmann, D. (1973). A n d . Chem. 45, 2270. Liu, Y. Y., Schmeltz, I., and Hoffmann, D. (1974). Anal. Chein. 46, 885-889. Magee, P. N., and Barnes, J. M. (1956). Br. J . Cancer 10, 114. Matht, D., Schweisguth, O., Schneider, M., Amiel, J . L., Berumen, Z., Brule, G., Cattan, A., and Schwarzenberg, L. (1963). Lnncet 2, 1077-1080. Matsumoto, K., Mori, K., and Jasuno, A. (1960). Gann 51, 91. Mori, K., and Jasuno, A. (1959). Gann 50, 107- 110. Mori, K., Jasuno, A., and Matsumoto, K. (1960). Gan/i 51, 83-89. Oswald, H., and Kriiger, F. W. (1969). Arzneimittel Forsch. 19, 1891-1892. Peacock, A., and Peacock, P. R. (1966). Br. J . Cancer 20, 307-325. Poduschka, R. (1900). Arch. Exp. Puthol. Phunn. 44, 59-67. Pohl, J. (1902). Arch. Exp. Pathol. Pharm. 48, 367-375. Pompe, K. (1956). Derin. Wschr. 133, 105-108. Reddy, B. S . , Weisberger, J . N., Narisawa, T., and Wynder, E. E. (1974). Cancer R r s . 34, 2368-2372. Roe, F. T . C., Boyland, E., and Haddow, A. (1965). Br. Med. J . 1, 1550. Roe, F. T. C., Grant, G. A., and Milican, D. M. (1967). Ncrfure (Lo,idon) 216, 375-376. Rutishauser, A., and Bollag, W. (1967). Experientiu 23, 222-223. Schmahl, D., Preussmann, R., and Hamperl, H. (1960). Naturwis.senschrrfte/i 47, 89- 196. Schmahl, D., Danisman, A., Habs, M., and Diehl, B. (1976). Z. Krehsforsch. Klin. Onkol. 86, 89-94. Schwan, S. (1961). Prrthol. Pol. 12, 53. Schwan, S. (1962). Pfithol. Pol. 13, 185. Severi, L., and Biancifiori, C. (1968). J . N u t / . Caricer Inst. 41, 331-340. Tan, M. H., Holyoke, E. D., and Goldrosen, M. H. (1976). J. Nor/. Cancer Inst. 56, 871873. Thomas, C. (1965). Z. Krehsforsch. 67, 1-30. Thomas, C., and Kersting, G. (1964). Nuturwissenschajien 51, 144- 145. Thurnherr, N., Deschner, E. E., Stonehill, E. H., and Lipkin, M. (1973). Cuncer Res. 33, 940- 945. Toth, B. (1975). Cancer Res. 35, 3693-3697. Toth, B. (1976). Cancer R e s . 36, 917-921. Toth, B., and Wilson, R. B. (1971). A m . J. Puth. 64, 585-600. Troncale, F., Hertz, R., and Lipkin, M. (1971). Cancer Res. 31, 463-467. Underhill, F., and Kleiner, I. S. (1908). J . B i d . Chern. 4, 165. Viallier, J., and Casanova, F. (1960). C . R . S . B i d . (Paris) 154, 985-987. Wagner, H., and Moritz, R. (1962). Arch. Geschwulstforsch. 19, 123- 129. Weinstein, H. J., and Kinosita, R. (1962). J. Lab. Clirz. Med. 60, 1025. Weitzel, G., Schneider, F., Fretzdorf, A. M., Seynsche, K., and Finger, H. (1963). Z. Physiol. Chern. 334, 1-25.
164
JOSEPH BAL6
Wells, H. G . (1908). J . B i d . Chem. 4, 165. Wells, H . G . (1908). J . E.rp. Med. 10, 457-464. Wiebecke, B., Lohrs, V., Girnmy, .I. and , Eder, M. (1969). Z. Ges. Exp. Meci. 149, 277278. Wolfart, W. (1960). Dtsch. Mecl. Wschr. 85, 1655- 1657. Zeller, P., Gutmann. H . , Hegedus, B., Kaiser, A . , Langemann, A., and Muller, M. (1963). Experientiri 19, 129.