Tumorigenicityof Bay-Regionepoxidesand Other Derivativesof Chrysene and Phenanthrenein Newbornmice

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[CANCERRESEARCH39,5063-5068,December1979] 0008-5472/79/0039-0000502.00 Tumorigenicityof Bay-RegionEpoxideSand Other Derivativesof Chrysene and Phenanthrenein NewbornMice

Mildred K. Buening, Wayne Levin,1 Jean M. Karle, Haruhiko Yagi, Donald M. Jerina, and Allan H. Conney

Department of Biochemistry and Drug Metabolism, Hoffmann-La Roche Inc., Nutley, New Jersey 071 10 (M. K. B., W. L., A. H. C.], and Laboratory of Bioorganic Chemistry, National Institute of Arthritis, Metabolism, and Digestive Diseases, NIH, Bethesda, Maryland 20205 [J. M. K., H. Y., D. M. J.j

ABSTRACT nogenic activity (4). Chrysene, which has one more benzene ring than does phenanthrene, is a symmetrical molecule with The tumorigenic activities of several derivatives of chrysene 2 identical bay regions (Chart 1). Chrysene has weak activity and phenanthrene were tested in newborn Swiss-Webster in carcinogenicity tests (4, 11). Quantum mechanical aspects mice. The compounds were administered i.p. in doses of 0.2, of the bay region theory predict (6) that bay-region 1,2-diol 0.4, and 0.8 @tmolonthe first, eighth, and 15th days of life. 3,4-epoxides of chrysene and phenanthrene should have sim Experiments were terminated when the animals were 38 to 42 ilar chemical reactivities. Kinetic studies of their solvolysis in weeks old. The only tumors found in control mice were lung water from pH 2 to 10 have established that this is indeed the adenomas; 15% of the control animals developed these pul case (19). Since the calculations also predict that these 1,2- monary tumors with an average of 0.1 7 tumor/mouse. (Â±)- diol-3,4-epoxides should be the most reactive of the possible 1fi, 2a-Dihydroxy-3a ,4a-epoxy- 1,2,3,4-tetrahydrochmysene isomemicbenzo-ningdiol-epoxides of the 2 hydrocarbons, these (diol epoxide-2) with the benzylic 1-hydroxyl group trans to the diol-epoxides and their dihydrodiol precursors are prime can bay-region epoxide oxygen was the most potent chrysene didates as ultimate and proximate carcinogens, respectively derivative tested. It induced pulmonary tumors in 98% of the (5, 6). mice, with an average of 15.9 tumors/mouse. (Â±)-lfl,2a-Di In a bacterial mutagenicity test, chrysene 1,2-dihydmodiol3 hydroxy-3/3,4fl-epoxy-1 ,2,3,4-tetrahydmochmysene(diol epox was metabolically activated by hepatic enzymes to products ide-i ), in which the bay-region epoxide oxygen is cis to the that were 20 times more mutagenic to strain TA 100 of Sal benzylic 1-hydroxyl group, had little if any tumorigenic activity monella typhimurium than were the metabolites formed from at the dose tested. trans-i ,2-Dihydroxy-i ,2-dihydmochrysene chmysene,chrysene 3,4-dihydrodiol, or chrysene 5,6-dihydro and 1,2-dihydrochrysene, the immediate metabolic precursors diol (24). When the double bond in the 3,4-position of chrysene of a bay-region diol-epoxide and a bay-region tetmahydmoepox 1,2-dihydrodiolwassaturated,theresultingtetrahydrodiol ide, respectively, were the next most active chmysenederiva could not be metabolically activated to mutagenic metabolites. tives tested; they produced pulmonary tumors in 73 to 75% of In a study of the tumor-initiating activities of chrysene and its the mice, with an average of 2. 1 to 2.2 tumors/mouse. 3,4- 3 metabolically possible dihydmodiolson mouse skin, chrysene Epoxy-i ,2,3,4-tetrahydrochrysene (bay-region epoxide) in 3,4- and 5,6-dihydrodiol had no significanttumorigenic activity, duced pulmonary tumors in 71% of the mice, with an average but chrysene 1,2-dihydrodiol had about twice the tumonigenic of 1.26 tumors/mouse. trans-3,4-Dihydroxy-3,4-dihydrochmy activity of the parent hydrocarbon (11). When the double bond sene, 3,4-dihydrochrysene, and trans-5,6-dihydmoxy-5,6-di in the 3,4-positionof chrysene1,2-dihydrodiolwas saturated, hydrochrysene had little or no tumorigenic activity. Chrysene the resulting tetrahydrodiol had less than 25% of the tumomi itself had little or no tumorigenic activity in the lung but pro genic activity of chrysene 1,2-dihydrodiol. These results mdi duced a 25% incidence of hepatic tumors in male mice, with cate that chrysene 1,2-dihydrodiol is a proximate carcinogenic an average of 0.42 tumor/mouse. Chrysene derivatives which metabolite and suggest that the 3,4-position of the molecule is induced a large number of pulmonary tumors also induced a critical site for further metabolism to an ultimate carcinogenic some hepatic tumors and lymphomas. Phenanthrene and its metabolite. derivatives were less tumorigenic than chrysene and its deny atives. Phenanthrene, 1,2-dihydrophenanthrene, and the two diastereomenic bay-region diol-epoxides of phenanthrene had region is the sterically hindered region between the 4- and 5-positions of little or no tumorigenic activity, but the bay-region tetrahydro phenanthrene. The bay regions in chrysene are between carbon atoms 4 and 5 epoxide, 3,4-epoxy-i ,2,3,4-tetrahydrophenanthrene, had and between carbon atoms 10 and 11. Due to the symmetry of chrysene, these 2 bay regions are identical (Chart 1). some activity. This compound induced pulmonary tumors in 3 The abbreviations used are: chrysene 1 ,2-dihydrodiol, trans-i ,2-dihydroxy 45% of the mice, with an average of 0.74 tumor/mouse. It also 1,2-dihydrochrysene; chrysene 3,4- and 5,6-dihydrodiol, other transdihydrodiols of chrysene; 1 2-H2 chrysene, 1,2-dihydrochrysene; 3,4-H2 chrysene, 3,4-dihy induced a few hepatic tumors and lymphomas. drochrysene; chrysene H4-3,4-epoxide, 3,4-epoxy-i ,2,3,4-tetrahydrochrysene; chrysene 1,2-diol-3,4-epoxide-1, (Â±)-lfI,2a-dihydroxy-3f1,4$-epoxy-1,2,3,4-tet INTRODUCTION rahydrochrysene; chrysene 1,2-diol-3,4-epoxide-2, (Â±)-i$,2a-dihydroxy-3a,4a- epoxy-i ,2,3,4-tetrahydrochrysene; i 2-H2 phenanthrene, 1,2-dihydrophenan threne; phenanthrene H4-3,4-epoxide, 3,4-epoxy-i ,2,3,4-tetrahydrophenan Phenanthrene, the simplest polycyclic aromatic hydrocarbon threne; phenanthrene 1,2-dlol-3,4-epoxide-i , (Â±)-i @,2a-dihydroxy-3fl,4fl with a bay region,2 is generally considered to have no carci epoxy-i ,2,3,4-tetrahydrophenanthrene; phenanthrene 1,2-diol-3,4-epoxide-2, (Â±)-ifl,2a-dihydroxy-3a,4a-epoxy-i ,2,3,4-tetrahydrophenanthrene; benzo(a) pyrene 7,8-dlol-9,1 0-epoxide, one or more of the four isomers of the dia ReceivedJuly6, 1979;acceptedSeptember13, 1979 stereomeric 9.10-epoxides derived from (+)- or (â€”)-trans-7,8-dihydroxy-7,8- 1 To whom requests for reprints should be addressed. dihydrobenzo(a)pyrene, in which the epoxide is cis (diol-epoxide-1 ) or trans (diol 2 A bay region is present in a polycyclic aromatic hydrocarbon when an epoxide-2) to the benzylic 7-hydroxyl group; DMSO, dimethyl sulfoxide. Where angular benzo ring is present in the molecule. The simplest example of a bay enantiomers are possible, racemic mixtures were used in the present study.

DECEMBER1979 5063

@ OM..,OH BLU:Ha (ICR) strain were obtained from Blue Spruce Farms, @ 0 0 @Iecl@POH Altamont, N. V., 1 to 9 days before partumition.Within 24 hr of

OH @H birth, 1 0 pups in each litter were given i.p. injections of the first

CHRYSENE CHRYSENE CHRYSENE dose of compound. The mice were given a total dose of 1.4 .2- DIPIYDRODIOL 34. DIHYDRODIOL 5,6-DINYDRODIOI...... Bay @zmolofcompound dIvIded Into 3 InjectIons of 0.2, 0.4, and 0.8 @tmoI,administeredon the first, eighth, and 15th days of life. @ o:9@@ cx9@ ControlmiceweregiveninjectionsofDMSO.Themicewere :@.â€˜ 0 0 0 0 0 housed in plastic cages with corncob bedding and were fed R.g*on ,2.H@ CNRYStNE 3,4'H2 CHRYSENE CHRYSENE Purina laboratory chow (Ralston Purina Co. St. Louis Mo.) ad H4 @3,4-EPOXlDE CHRYSENE libitum. The mice were weaned at 23 days of age, and the OH Oh experiment was terminated by killing the animals when they 0 â€˜@ 0 : were 38 to 42 weeks old. At autopsy, the major organs of each 0 0 0 0 0 -@5 animal were examined grossly, tumors were counted, and

CHRYSENE 1,2-DIOL- CHRYSENE 1,2-DIOL- tissues were fixed in 1 0% buffered formalin. A representative

3,4-EPOXIDE-I 3,4-EPOXIDE-2 number of pulmonary tumors, all hepatic tumors, and all other tissues with suspected pathology were examined histologically. 0 ..- 0 Pathology of the lung tumors was the same as has been 0 0 0 described previously (13).

1,2-H2 PHENANTHR(N( PHENANTHRENE H4'3,4-EPOXIDE RESULTS

Because of the weak tumonigenic activity of chrysene, we PHENANTHRENE @,,OH @1:@oH administered the maximum dose that could be given to newborn mice with our experimental protocol. The dose was limited by PHENANTHRENE 1,2-DIOL- PHENANTHRENE the solubility of chrysene in DMSO and by the toxicity of DMSO 3,4-EPOXIDE-i 3,4-EPOxIDE-2 to the newborn mice. The limit of solubility of chrysene in Chart 1. Structures of chrysene, phenanthrene, and their derivatives, tested DMSO is about 20 mM, and we therefore doubled the volume for tumorigenic activity in newborn mice. Stereochemistry is relative. of the injections from the amount used in previous experiments (2, 7, 21). Although the phenanthrene derivatives are more In recent studies (23), the diastereomenic 1,2-diol-3,4-epox soluble in DMSO, we chose to use a dose equimolamto the ides of chrysene were 5 to 60 times more mutagenic than was chrysene derivatives so that the biological activities of the 2 the K-region chrysene 5,6-oxide in strains TA 98 and TA 100 hydrocarbons could be compared directly. The survival at of S. typhimurium and in Chinese hamster V79 cells. The weaning of control mice given i.p. injections of 10, 20, and 40 diastemeomeric 1,2-diol-3,4-epoxides of phenanthmene ex z1of DMSO on the first, eighth, and 15th days of life was 66%. hibited dose-dependent mutagenic activity in bacterial cells but PhenanthmeneH4-3,4-epoxide had the highest acute toxicity. were less mutagenic than were the corresponding diol-epox From the initial group of 140 mice, composed of 14 litters of ides of chrysene. The bay-region tetrahydroepoxides, chry 10 miceeach,only63%survivedtotheeighthday.However, sene H4-3,4-epoxide and phenanthrene H4-3,4-epoxide, were 1,2-H2 chrysene and 1,2-H2 phenanthrene showed toxicity more mutagenic than were their corresponding bay-region diol equal to phenanthrene H4-3,4-epoxide when the animals were epoxides in both bacterial and mammalian cells. The present weaned at 23 days of age. At the termination of the study (38 study was undertaken to test the tumomigenic activity of the to 42 weeks of age), 1,2-H2 phenanthrene was the most toxic bay-region diol-epoxides and tetmahydroepoxides of chrysene compound investigated, with a mean survival of only 17%. The and phenanthrene in newborn mice. Benzo-ning dihydro deny third injection of chrysene 3,4-dihydrodiol anesthetized all the atives of the hydrocarbons were also tested, since they can be animals, but these animals recovered, and the survival at metabolized to tetmahydroepoxides. weaning in this group was 53%. The tumonigenic activity of chrysene, phenanthrene, and 12 MATERIALS AND METHODS of their derivatives in newborn mice is shown in Table 1. In the Polycyclic Hydrocarbons and Their Derivatives. Chrysene control group, 15% of the mice developed pulmonary tumors, (95% pure; m.p. 250â€”252Â°)and phenanthrene (>98% pure) with an average of 0.1 7 tumor/mouse; and in mice treated with were purchased from Aldrich Chemical Co., Milwaukee, Wis. chrysene, only 17% developed pulmonary tumors, with an Chrysene, after recrystallization 5 times from benzene (m.p. average of 0.34 tumor/mouse. Chrysene 1,2-dihydrodiol in 256Â°), and phenanthmene, as supplied, were homogeneous on duced pulmonary tumors in 73% of the mice, with an average reverse-phase high-pressure liquid chromatography. All chry of 2.07 tumors/mouse, and it was much more active than the sene and phenanthrene derivatives, except the diol-epoxides, other metabolically possible trans-dihydrodiols of chrysene. were prepared as described (8, 9), and all the compounds Less than 25% of mice treated with chrysene 3,4- or 5,6- were free of impurities, as determined by nuclear magnetic dihydrodiol developed pulmonary tumors, with an average of resonance and mass spectrometry. Diol-epoxide 2 isomers less than 0.30 tumor/mouse. 1,2-H2 Chmysenewas equal to were obtained by direct epoxidation and diol-epoxide 1 isomers chrysene 1,2-dihydrodiol in tumonigenic activity; it induced were obtained from intermediate bromotniols as described (19). pulmonary tumors in 75% of the mice, with an average of 2.22 The compounds were stored at â€”90Â°.Forinjections into mice, tumors/mouse. 3,4-H2 chrysene had little or no tumonigenic 20 m@solutions in anhydmousDM50 were prepared. activity. Tumorigenicity Study. Pregnant mice of the Swiss-Webster The diastereomenic chrysene 1,2-diol-3,4-epoxides differed

5064 CANCER RESEARCH VOL. 39

Table miceSwiss-WebsterTumorigenicity of derivatives of chrysene and phenanthrene in newborn mice [BLU:Ha (ICR)] were given i.p. injections on the first, eighth, and 15th day of life with 0.2, 0.4, and 0.8 @.tmolofcompound, respectively. When the experiment was terminated, the major organs of each animal were examined, tumors were counted, and tissues were fixed in formalin. A representative number of pulmonary tumors, all hepatic tumors, and all other tissues with suspected pathology were examinedhistologically.Pulmonary tu No. of mice alive mors Hepatic tumorsa % of mice At termi- % of % of with At wean- nation, mice Av. mice Av. other In- ing, Day 38â€”42 with tu- no.! with tu- no.! neo jectedplasms100 23 wk mors mouse mors mouse Control Female 39 38 18 0.21 0 0 0 Male 27 21 10 0.10 0 0 0 Total 66 59 15 0.17

Chrysene 100 Female 24 11 9 0.18 0 0 0 Male 30 24 21 0.42 25 0.42 Total 54 35 17 0.34

Chrysene 1,2-dihydrodiol 100 Female 32 32 88 2.59 0 0 Male 33 24 54 1.37 46 0.67 Total 65 56 73 2.07

Chrysene 3,4-dihydrodiol 100 Female 19 18 6 0.11 0 0 6â€• Male 34 27 18 0.22 0 0 0 Total 53 45 13 0.18

Chrysene 5,6-dihydrodiol 80 Female 25 22 23 0.28 0 0 Male 31 26 23 0.27 0 0 0 Total 56 48 23 0.27

1,2-H2chrysene 100 Female 13 12 67 3.25 0 0 25e Male 27 20 80 1.60 20 0.35 5'@ Total 40 32 75 2.22

3,4-H2chrysene 100 Female 29 26 31 0.42 0 0 4d Male 36 30 26 0.29 10 0.10 3' Total 65 56 28 0.35

Chrysene H4-3,4-epoxide 100 Female 20 16 56 1.12 0 0 Male 31 26 81 1.31 38 0.58 0 Total 51 42 71 1.24

Chrysene 1,2-diol-3,4-epoxide-1 100 Female 31 31 32 0.32 0 0 0 Male 30 25 24 0.36 8 0.08 0 Total 61 56 29 0.34

Chrysene 1,2-diol-3,4-epoxide-2 100 Female 29 24 96 15.68 4 0.04 17â€• Male 37 23 100 16.78 39 0.87 Total 66 47 98 15.91

Phenanthrene 100 Female 26 18 6 0.11 0 0 6@' Male 24 17 30 0.30 0 0 0 Total So 35 17 0.20

1.2-H2 phenanthrene 100 Female 16 7 14 0.14 0 0 0 Male 25 10 10 0.10 10 0.10 0 Total 41 17 12 0.12

Phenanthrene H,-3,4-epoxide 140 Female 28 20 58 0.95 0 0 10â€• Male 26 27 41 0.60 15 0.19 4b Total 54 47 45 0.74

DECEMBER1979 5065

â€”continuedNo.Table 1 tu of mice alivePulmonary morsHepatic tumorsa% of mice ofAt withAttermi % of% Av.otherIn-wean- nation,mice Av.mice ing, Day 38â€”42with tu- no.!with tu- no.!neo mouseplasmsPhenanthrene1 jected 23 wkmors mousemors 100ide-iFemale21,2-diol-3,4-epox

05dMale39 2020 0.250 00Total60 3010 0.130 0.18Phenanthrene1 5014

100ide-2Female21,2-diol-3,4-epox

07CMale27 1421 0.210 0.094â€•Total48 2313 0.139 3716 0.16

a Except for one reticulum cell sarcoma in a female mouse treated with chrysene 1 ,2-diol-3,4-epoxide-2, all hepatic tumors were type A nodules (18, 20). b Splenic lymphosarcoma; localized. CThymiclymphosarcoma;localized. d Malignant lymphoma; disseminated. e Malignant lymphoma; 1 localized; 2 disseminated. f Duodenal adenocarcinoma. ppLymphosarcoma:g Malignantlymphoma;3localized:1disseminated.disseminated.

Table 2 Statistical analysis of pulmonary and hepatic tumor incidence in newborn mice treated with chrysene derivatives Ten treatment groups were compared for tumor incidence by Fishers exact test for proportions (1). ChryseneChryseneChryseneChryseneChrysene3,4-Dihy Chrysene1 ,2-diol 1,2-diol 1,2-dihy 3,4-dihy 5,6-dihy 1.2-H2drochry H4-3,4-3,4-epox 3,4-epox 2Controlâ€”HaP.TreatmentControlChrysenedrodioldrodioldrodiolChryseneseneepoxideide-iide HChryseneâ€”PHHPPPChrysene HP. HP. HP.

HPChrysene1,2-dihydrodiolâ€”P. HP. HP, HP, HChrysene3,4-dihydrodiolâ€”P. HP. HP. H1 5,6-dihydrodiolâ€”P. HP. HP. .2-H2chryseneâ€”PPP3,4-H2 HChrysenechryseneâ€”P. HP. HPChryseneH,-3,4-epoxideâ€”P. Hepoxide-iChrysene1,2-diol-3,4-â€”P.

1,2-diol-3,4-â€”epoxide-2

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Table3 Statistical analysis of pulmonary and hepatic tumor incidence in newborn micederivativesPhenan- treated with phenanthre Six treatment groups were compared for tumor incidence by Fisher's exact test for proportions (i).ne Phenan Phenan threnethrenePhenan- threne ,2-diol-3,4-Treatment 1.2-H2 phen- H4-3,4- 1,2-diol-3,4-1 epoxide-iepoxide-2Control Control threne anthrene epoxide paPhenanthrene â€” pi â€” pPhenanthrene,2-H2 phenanthrene â€” HPPhenanthreneH4-3,4-epoxide â€” P. â€”epoxide-1Phenanthrenei ,2-diol-3,4-

,2-diol-3,4-â€”epoxlde-21

a A statistically significant incidence of tumors (p < 0.05) is designated by P for pulmonary tumors and H for hepatic tumors. The lack of any notation indicates no significant differences ( p > 0.05) between groups for either pulmonary or hepatic tumors. that chrysene 1,2-dihydrodiol is a proximate carcinogenic me interactions with critical target sites than when chrysene H4- tabolite of chrysene (1 1). The present results also demonstrate 3,4-epoxide itself is administered. that a bay-region diol-epoxide is an ultimate carcinogen of In newborn mice, little or no tumonigenic activity was ob chrysene in newborn mice. Although chrysene 1,2-diol-3,4- served with phenanthrene, the 1,2-dihydro derivative of phen epoxide-2 (epoxide trans to the benzylic 1-hydroxyl group) is anthrene, or the diastereomers of the bay-region diol-epoxide highly tumorigenic in the newborn mouse, chrysene 1,2-diol of phenanthrene. However, significant tumonigenic activity was 3,4-epoxide-1 (epoxide cis to the benzylic 1-hydnoxyl group) observed with the bay-region tetrahydroepoxide, phenanthrene had little or no activity. In previous studies of the tumomigenicity H4-3,4-epoxide (Table 1). These results are consistent with of the diastereomeric bay-region diol-epoxides of both previous results indicating that phenanthrene H4-3,4-epoxide benzo(a)pyrene and benzo(a)anthracene in newborn mice, has significant tumomigenicactivity on mouse skin and muta diol-epoxide-2 isomers induced many more pulmonary tumors genic activity in S. typhimurium and in V79 cells (23). In than did the parent hydrocarbon, whereas diol-epoxide-i iso mutagenicity and tumomigenicitystudies on mouse skin, phen mers had little or no tumonigenic activity (2, 7, 21). In tumoni anthmene H4-3,4-epoxide had 28 to 61% of the activity of genicity studies on mouse skin, diol-epoxide-2 isomers were chrysene H4-3,4-epoxide, and in newborn mice phenanthrene also more active than were corresponding diol-epoxide-1 iso H4-3,4-epoxide was 60% as tumonigenic as was chrysene H4- mers, although benzo(a)pyrene was more active than was its 3,4-epoxide (23). Compared to the corresponding bay-region bay-region diol-epoxide-2 isomer (10, 12, 15, 16). diol-epoxides of chrysene, benzo(a)pyrene (2, 7), and When the average number of pulmonary tumors per mouse benzo(a)anthracene (21), the bay-region diol-epoxides of are used for comparison of tumonigenic activities, chrysene phenanthrene have little or no tumonigenic activity in newborn i ,2-diol-3,4-epoxide-2 was about 13 times more active than mice. Although the diol-epoxides and tetrahydroepoxides of was the corresponding bay-region tetrahydroepoxide, chry phenanthrene have chemical reactivity similar to that of the sene H4-3,4-epoxide. In mutagenicity studies with bacterial and corresponding chrysene derivatives (19), the phenanthrene mammalian cells, chrysene H4-3,4-epoxide was 1.4- to 6-fold derivatives are far less tumonigenic. The inability of the phen more active than was chrysene 1,2-diol-3,4-epoxide-2 (23). anthrene bay-region diol-epoxide or the tetmahydmoepoxideto The tumorigenic activity of the bay-region tetmahydroepoxide reach and react with critical target sites, and/or the inability of of chrysene may be less than that of chrysene 1,2-diol-3,4- the bound target sites to initiate the carcinogenic process, may epoxide-2 because of the action of epoxide hydrase on the contribute to the poor tumorigenic activity of phenanthrene. In tetrahydroepoxide. Benzo-ming diol-epoxides of benzo(a) addition, metabolic factors may also contribute to the lack of pyrene Ci7, 25), benzo(a)anthracene(22), chrysene(23), and carcmnogenicity of phenanthrene. Although phenanthrene is phenanthrene (23) are refractory to hydration by epoxide hy metabolized to the 1,2-dihydrodiol by matliver microsomes (3, drase, but mutagenic arene oxides and tetmahydmoepoxidesof 14), it is not known whether significant amounts of a bay-region these hydrocarbons are readily inactivated by this enzyme. diol-epoxide are formed from the dihydrodiol. When test compounds are administered i.p. to newborn mice, We conclude that the carcinogenicity of chrysene follows the a major portion of the dose passes through the liver, and the prediction of the bay-region theory (5, 6) and that chrysene is compounds are available to metabolizing enzymes in this organ metabolized to chmysene 1,2-dihydrodiol, a proximate carci prior to reaching the lung. As a consequence, it is possible that nogenic metabolite, and then to a bay-region diol-epoxide more of the reactive diol-epoxides than of the tetrahydmoepox which is an ultimate carcinogenic metabolite. Although molec ides will reach critical target sites in the lung. The susceptibility ular orbital calculations indicate similar chemical meactivities of chrysene H4-3,4-epoxide to inactivation by hepatic epoxide for the bay-region epoxides of chrysene (i@Ede@/$ 0.640) hydrase (23) may also explain why 1,2-H2 chrysene, the po and phenanthrene (h@Edeyoc/$ 0.658) (6), the bay-region diol tential metabolic precursor of chrysene H4-3,4-epoxide, in epoxides of phenanthrene had little or no tumonigenic activity duces more pulmonary tumors than does chrysene H4-3,4- in newborn mice. However, the less polar bay-region tetrahy epoxide. When 1,2-H2chrysene is administered, its distribution droepoxide of phenanthrene had significant tumonigenic activ and metabolism may result in the in vivo formation of chrysene ity in the newborn mouse. Phenanthrene itself, the simplest H4-3,4-epoxide at locations which allow a greater number of polycyclic aromatic hydrocarbon with a bay region, lacks sig

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Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 1979 American Association for Cancer Research. M. K. Buening et a!. nificant carcinogenic activity in all tumor models tested. Per ide is an ultimate carcinogen on mouse skin. Cancer Res., 38: 1705â€”1710, haps the small size and resulting increased polarity of the 1978. 11. Levin, W., Wood, A. W., Chang, R. L.. Yagi, H., Mah, H. 0., Jerina, D. M., phenanthrene diol-epoxides and tetmahydroepoxides limit their and Conney, A. H. Evidence for bay region activation of chrysene 1.2- intrinsic activity, particularly if intercalation of the hydrophobic dihydrodiol to an ultimate carcinogen. Cancer Res., 38: 1831 â€”1834,1978. planar portion of the hydrocarbon into the DNA base stack 12. Levin, W., Wood, A. W., Wislocki, P. G., Kapitulnik, J., Yagi, H., Jerina, D. M., and Conney, A. H. Carcinogenicityof benzo-ringderivativesof must accompany covalent binding at the critical target site to benzo(a)pyrene on mouse skin. Cancer Res., 37: 3356â€”3361,1971. initiate the carcinogenic process. 13. Shimkin, M. B., and Stoner, G. D. Lung tumors in mice: application to carcinogenesis bioassay. Adv. Cancer Res., 21: 1â€”58,1975. 14. Sims, P. Qualitative and quantitative studies on the metabolism of a series ACKNOWLEDGMENTS of aromatic hydrocarbons by rat-liver preparations. Biochem. Pharmacol., 19: 795-818, 1970. We thank Nelson Montero for his help in caring for the animals and Ann Marie 15. Slaga, T. J., Bracken, W. M., Gleason, G., Levin, W., Vagi, H., Jerina, 0. M., Willams for her assistance in the preparation of this manuscript. We thank Dr. and Conney, A. H. Marked differences in the skin tumor-initiating activities Gary Williams, C. 0. Wong, and the staff of the Naylor Dana Institute for Disease of the optical enantiomers of the diastereomeric benzo(a)pyrene 7,8-diol Prevention, American Health Foundation, Valhalla, N. V., for the histological 9,10-epoxides. Cancer Res.. 39: 67â€”71,1979. studies. 16. Slaga, T. J., Bracken, W. M., Viaje, A., Levin, W., Yagi, H., Jerina, D. M., and Conney, A. H. Comparison of the tumor-initiating activities of benzo(a] pyrene arene oxides and diol epoxides. Cancer Res., 3 7: 4130â€”4133, 1977. REFERENCES 17. Thakker, D. R., Yagi, H., Lu, A. V. H., Levin, W., Conney, A. H., and Jerina, D. M. Metabolism of benzo(a)pyrene. IV. Conversion of (Â±)-trans-7,8-dihy i . Bradley, J. V. Distribution-Free Statistical Tests, pp. 195â€”203.Englewood droxy-7,8-dihydrobenzo(a)pyrene to the highly mutagenic 7,8-diol-9, 10- Cliffs, N. J.: Prentice-Hall, 1968. epoxides. Proc. NatI. Acad. Sci. U. S. A., 73: 3381 -3385, 1976. 2. Buening. M. K., Wislocki, P. G.. Levin, W., Yagi, H., Thakker, 0. R., Akagi, 18. Walker, A. I. T., Thorpe, E., and Stevenson, D. E. The toxicity of dieldrin H., Koreeda, M., Jerina, D. M., and Conney, A. H. Tumorigenicity of the (HEOD). I. Long-term oral toxicity studies in mice. Food Cosmet. 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Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 1979 American Association for Cancer Research. Tumorigenicity of Bay-Region Epoxides and Other Derivatives of Chrysene and Phenanthrene in Newborn Mice

Mildred K. Buening, Wayne Levin, Jean M. Karle, et al.

Cancer Res 1979;39:5063-5068.

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