genotoxicity and carcinogenicity testing of pharmaceuticals: correlations between induction of dna...

Mutation Research 705 (2010) 20–39

Review

Genotoxicity and carcinogenicity testing of pharmaceuticals: Correlationsbetween induction of DNA lesions and carcinogenic activity

Giovanni Brambilla *, Francesca Mattioli, Luigi Robbiano, Antonietta Martelli

Department of Internal Medicine, Division of Clinical Pharmacology and Toxicology, University of Genoa, Viale Benedetto XV, 2, I-16132 Genoa, Italy

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

2. Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

3. Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36

References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

A R T I C L E I N F O

Article history:

Received 22 January 2010

Accepted 6 February 2010

Available online 3 March 2010

Keywords:

Pharmaceuticals

DNA adducts

DNA strand breaks

DNA repair synthesis

Carcinogenicity

A B S T R A C T

This survey is a compendium of the results of DNA lesions assays (DNA adducts, DNA strand breaks, DNA

repair synthesis) and of the results of carcinogenicity assays of 146 pharmaceuticals. Of these drugs, 55

(37.7%) tested negative in both DNA lesions assay(s) and in carcinogenicity assay(s); 65 (44.5%) tested

negative in DNA lesions assay(s), but gave a positive response in at least one carcinogenicity assay; 6

(4.1%) tested positive in at least one DNA lesions assay, but negative in carcinogenicity assay(s); 20

(13.7%) tested positive in at least one DNA lesions assay and in at least one carcinogenicity assay.

Concerning the predictivity of DNA lesions assays findings for the results of long-term carcinogenesis

assays performed in mice, rats or other species, concordance was found to exist for the 46.2% of

pharmaceuticals in the case of DNA adducts, for 63.1% in the case of DNA strand breaks, and for 47.3% in

the case of DNA repair synthesis (UDS).

� 2010 Elsevier B.V. All rights reserved.

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1. Introduction

Present guidelines for genotoxicity studies of pharmaceuticals[1–3] suggest that, in addition to the standard 3-test battery, it isuseful to perform also assays on the possible occurrence of DNAlesions, i.e. tests for assessing the induction of DNA strand breaksand cross-links, of DNA repair synthesis (UDS), and of theformation of DNA adducts. Therefore we deemed useful toexamine to what extent the results provided by DNA lesionsassays are positively correlated with the results of long-termcarcinogenicity assays. We have found 146 marketed pharmaceu-ticals that have been tested for both DNA-damaging andcarcinogenic activity. This review is a compendium of all the datathat have been found in an extensive search. For each drugconsidered the search terms were DNA lesions and carcinogenic

* Corresponding author at: DI.M.I., Sezione di Farmacologia, Universita di Genova,

Viale Benedetto XV, 2, I-16132 Genova, Italy. Fax: +39 010 353 8232.

E-mail address: [email protected] (G. Brambilla).

1383-5742/$ – see front matter � 2010 Elsevier B.V. All rights reserved.

doi:10.1016/j.mrrev.2010.02.004

activity. The search was conducted primarily in peer-reviewedjournals using Medline, Toxline, and the Registry of Toxic Effects ofChemicals Substances [4]. Additional unpublished data wereobtained from Micromedex and from the following websites:http://www.toxnet.nlm.nih.gov, http://www.ntp.server.niehs.nih.gov, http://www.potency.berkeley.edu, http://www.fda.gov/cder,http://www.scirus.com, http://www.inchem.org, http://www.updateusa.com, http://www.osha.gov. Concerning data that arenot published in peer-reviewed journals, in some cases the testswere conducted under the oversight of authoritative bodies, such asthe U.S. National Toxicology Program; in the other cases the data arethose reported by the Physician’s Desk Reference [5] or in the finalpackage insert approved by the Center for Drug Evaluation andResearch of the Food and Drug Administration [6]. Unfortunately,this additional unpublished information is often incomplete; inparticular, the results of DNA-damage assays are usually reportedwithout any information of the doses that have been tested.Moreover, often no information is given whether the in vitro assayswere performed in both the presence and the absence of anexogenous metabolic system; in these cases, taking into account that

http://www.toxnet.nlm.nih.gov/

http://www.ntp.server.niehs.nih.gov/

http://www.ntp.server.niehs.nih.gov/

http://www.potency.berkeley.edu/

http://www.fda.gov/cder

http://www.scirus.com/

http://www.inchem.org/

http://www.updateusa.com/

http://www.updateusa.com/

http://www.osha.gov/

mailto:[email protected]

http://www.sciencedirect.com/science/journal/13835742

http://dx.doi.org/10.1016/j.mrrev.2010.02.004

G. Brambilla et al. / Mutation Research 705 (2010) 20–39 21

this procedure is required by the guidelines, in the absence of aspecific indication the result is reported in the tables as obtained inboth these experimental conditions.

2. Results

Information on both DNA-damaging and carcinogenic activitywere retrieved for 146 marketed pharmaceuticals (Table 1)

Table 1DNA-damaging and carcinogenic activity of marketed pharmaceuticals.

Test system references Resultsa

Without e.m

1. Acarbose (56180-94-0)

Binding (covalent) to DNA in vitro �Long-term carcinogenesis assay, Sprague–Dawley rats + (kidney an

cell tumours

Long-term carcinogenesis assay, Sprague–Dawley rats �Long-term carcinogenesis assay, Wistar rats �Long-term carcinogenesis assay, hamsters �

2. Acemetacin (53164-05-9)

UDS, human lymphocyes in vitro �UDS, mouse spleen cells in vivo �Long-term carcinogenesis assay, rats �

3. Acitretin (55079-83-9)

UDS, rat primary hepatocytes �UDS, human fibroblasts in vitro �Long-term carcinogenesis assay, male mice + (vessels tu

Long-term carcinogenesis assay, female mice � (x 2.8)c

Long-term carcinogenesis assay, Wistar rats � (x 0.2)

4. Alendronate (66376-36-1)

DNA strand breaks, rat primary hepatocytes �Long-term carcinogenesis assay, male mice � (x 1.2)

Long-term carcinogenesis assay, female mice + (harderian

Long-term carcinogenesis assay, male rats + (thyroid ad

5. Alfuzosin (81403-80-7)

UDS, human cells in vitro �Long-term carcinogenesis assay, mice �Long-term carcinogenesis assay, rats �

6. Allopurinol (315-30-0)

Binding (covalent) to rat intestine DNA in vivo �Long-term carcinogenesis assay, mice � (x 0.1)

Long-term carcinogenesis assay, rats � (x 0.2)

7. Alosetron (122852-42-0)

UDS, rat hepatocytes in vivo �Long-term carcinogenesis assay, mice �Long-term carcinogeneis assay, rats �

8. Alprazolam (28981-97-7)

DNA strand breaks, animal cells in vitro �DNA strand breaks, rat hepatocytes in vivo �DNA strand breaks, rat hepatocytes in vivo �Long-term carcinogenesis assay, mice � (x 12)

Long-term carcinogenesis assay, rats �

9. Aripiprazole (129722-12-9)

UDS, rat hepatocytes in vivo �Long-term carcinogenesis assay, male ICR mice � (x 5.6)

Long-term carcinogenesis assay, female ICR mice + (pituitary a

tumours)

Long-term carcinogenesis assay, male F344 rats � (x 3.8)

Long-term carcinogeneis assay, female F344 rats + (mammary

Long-term carcinogenesis assay, male Sprague–Dawley rats � (x 22.8)

Long-term carcinogeneis assay,female Sprague–Dawley rats + (adrenal gl

10. Aspirin (50-78-2)

UDS, rat primary hepatocytes �Long-term carcinogenesis assay, CB6 mice � (x 0.5)



Long-term carcinogenesis assay, F334 rats � (x 0.5)

Long-term carcinogenesis assay, F344 rats � (x 0.04)

Long-term carcinogenesis assay, Wistar rats � (x 1.0)

[7–118]. As indicated by Table 2, the majority of drugs (55.5%)were examined only for the induction of DNA repair synthesis(UDS), in lower number (21.2%) only for the induction of DNAstrand breaks, and four (2.7%) only for the capability of formingDNA adducts. Taken as a whole only the 20.6% were examined bymore than one of the above mentioned assays. Concerningcarcinogenicity, the 82.9% of drugs were tested in both mice andrats and 8 of them also in other species, the 8.9% only in rats, the

Doseb

.s. With

e.m.s.

LED or HID

NT NR 5

d Leydig

)

500 mg/kg/day 5

NR 5

NR 5

NR 5

NT 10 ppm 8

4 mg/kg/day�6 months 9

120 mg/kg/day�3 months 9

NT NR 5

NT NR 5

mours) 0.5 MRHD/kg/day 5

0.5 MRHD/kg/day 5

2 mg/kg/day 5

NT NR 5

10 mg/kg/day 5

gl. tumours) 5 mg/kg/day 5

enomas) 3.75 mg/kg/day 5

NT NR 5, 6

100 mg/kg/day 5, 6

100 mg/kg/day 5, 6

50 mg/kg iv 5

20 mg/kg/day 5

20 mg/kg/day 5

NR 5

30 mg/kg/day 5

40 mg/kg/day 5

NT NR 5, 6

309 mg/kg po 10

65 mg/kg po�15 d 10

10 mg/kg/day 5, 6

30 mg/kg/day 5, 6

NR 6

30 mg/kg/day 6

nd mammary 3 mg/kg/day 6

10 mg/kg/day 6

tumours) 10 mg/kg/day 6

60 mg/kg/day 6

. tumours) 60 mg/kg/day 6

NT 1 mM 11

382 mg/kg/day 12

0.5% in diet 5

26.9 mg/kg/day 12

200 mg/kg/day 12

16 mg/kg/day 12

400 mg/kg/day 12

Table 1 (Continued )

Test system references Resultsa Doseb

Without e.m.s. With

e.m.s.

LED or HID

11. Atenolol (29122-68-7)

DNA strand breaks, Chinese hamster lung V79 cells in vitro � NT 799 mg/ml 13

DNA strand breaks, rat primary hepatocytes � NT 7990 mg/ml 14


UDS, rat primary hepatocytes � NT 799 mg/ml 13

DNA strand breaks, human primary hepatocytes � NT 7 99 mg/ml 13

UDS, human primary hepatocytes � NT 799 mg/ml 13

DNA strand breaks, rat hepatocytes in vivo � 75 mg/kg ip 14

Long-term carcinogenesis assay, C57BL/10J mice � (x 7.3) 300 mg/kg/day 15

Long-term carcinogenesis assay, Alderly Park rat � 300 mg/kg/day 16

Long-term carcinogenesis assay, male rats + (adrenal, pituitary,

and thyroid tumours)

500 mg/kg/day 16

Long-term carcinogenesis assay, female rats + (adrenal and

mammary tumours)

500 mg/kg/day 6

12. Atomoxetine (83015-26-3)

UDS, rat primary hepatocytes � NT NR 5

Long-term carcinogenesis assay, mice � 458 mg/kg/day 5

Long-term carcinogenesis assay, rats � (x 5.3) 47 mg/kg/day 5

13. Azelastine (58581-89-8)


Long-term carcinogenesis assay, mice � (x 15) 25 mg/kg/day 6

Long-term carcinogenesis assay, rats � 30 mg/kg/day 6

14. Bisoprolol (66722-44-9)




15. Bosentan (147536-97-8)

UDS, animal cells in vitro � NT NR 5

Long-term carcinogenesis assay, mice + (liver tumours) 450 mg/kg/day 5

Long-term carcinogenesis assay, rats + 500 mg/kg/day 5

16. Brotizolam (57801-81-7)

UDS, human embryonic fibroblasts in vitro � NT 300 mg/ml 17



17. Buspirone (36505-84-7)

DNA strand breaks, human Wi-38 cells in vitro � � NR 6

Long-term carcinogenesis assay, mice � 167 MRHD/kg/day 6

Long-term carcinogenesis assay, rats � 133 MRHD/kg/day 6

18. Butorphanol (42408-88-2)

UDS, human fibroblasts in vitro � � N R 6



19. Caffeine (58-08-2)

DNA strand breaks, Chinese hamster ovary cells in vitro + NT 388 mg/ml 18

DNA strand breaks, Chinese hamster lung

V79 cells in vitro

� NT 5800 mg/ml 18

DNA strand breaks, Syrian hamster embryo cells in vitro � NT 1000 mg/ml 18

UDS, Syrian hamster embryo cells in vitro � NT 1000 mg/ml 18

UDS, human lymphocytes in vitro � NT 388 mg/ml 18

DNA strand breaks, mouse liver and kidney in vivo + 100 mg/kg ip 18

Long-term carcinogenesis assay, female C3H mice + (mammary tumours) 500 mg/lt in dr.wt. 18

Long-term carcinogenesis assay, C57BL/6 mice � (x 0.4) 55 mg/kg/day 12

Long-term carcinogenesis assay, Wistar rats � (x 6.5) 0.2% i n dr.wt. 7, 18

Long-term carcinogenesis assay, Sprague–Dawley rats � (x 6.5) 0.2% in dr.wt. 7, 18

Long-term carcinogenesis assay, female Wistar rats + (pituitary adenomas) 0.2% in dr.wt 18

Long-term carcinogenesis assay, male

Sprague–Dawley rats

� (x 1.6) 0.102% in diet 18

Long-term carcinogenesis assay, Sprague–Dawley rats � (x 1.6) 100 mg/kg/day 18

20. Candesartan (139481-59-7)


Long-term carcinogenesis assay, mice � 100 mg/kg/day 5, 6

Long-term carcinogenesis assay, rats � 1000 mg/kg/day 5, 6

21. Carbidopa (28860-95-9)

UDS, rat primary hepatocytes � NT 0.5–50 mM 10

Long-term carcinogenesis assay, rats � (x 0.3) 2 MRHD/kg/day 5

22. Celiprolol (56980-93-9)

UDS, Chinese hamster lung V79 cells in vitro � � 3200 mg/ml 20

G. Brambilla et al. / Mutation Research 705 (2010) 20–3922



Without e.m.s. With

e.m.s.

LED or HID

Long-term carcinogenesis assay, CD-1 mice � (x 10.8) 900 mg/kg in diet 21

Long-term carcinogenesis assay, Sprague–Dawley rats � (x 19.5) 8000 ppm in diet 20

Long-term carcinogenesis assay, Sprague–Dawley rats � (x 21.9) 900 mg/kg in diet 20

Long-term carcinogenesis assay, beagle dogs � 300 mg/kg/dayn 20

23. Cerivastatin (143201-11-0)


Long-term carcinogenesis assay, mice + (liver tumours) 9.1 mg/kg/day 6

Long-term carcinogenesis assay, rats � (x 5.1) 0.158 mg/kg/day 6

24. Chlormadinone (1961-77-9)

UDS, rat primary hepatocytes � NT 50 mg/ml 22, 23

UDS, male rat primary hepatocytes � NT 20 mM 22, 24

UDS, female rat primary hepatocytes + NT 2 mM 22, 24

UDS, human primary hepatocytes + NT 5 mM 22, 24

Binding (covalent) to male rat hepatocytes DNA in vitro (+) NT 10 mM 22, 23

Binding (covalent) to female rat hepatocytes DNA in vitro + NT 10 mM 22, 23

Binding (covalent) to female rat hepatocytes DNA in vivo � 100 mg/kg po 22, 23

Long-term carcinogenesis assay, various strains of mice ? (x 1.9) 8 mg/kg in diet 25

Long-term carcinogenesis assay, CF-LP mice � 200–400 MRHD/kg/day 25

Long term carcinogenesis assay, rats � 400 MRHD/kg/day 25

Long term carcinogenesis assay, female beagle dogs + NR 25

25. Chlorphenamine (132-22-9)

DNA strand breaks, rat primary hepatocytes + NT 0.5 mM 26

UDS, rat primary hepatocytes � NT 0.5 mM 27, 28

Long-term carcinogenesis assay, male B6C3F1 mice � (x 23) 50 mg/kg/day 29

Long-term carcinogenesis assay, female B6C3F1 mice ? 200 mg/kg/day 29

Long-term carcinogenesis assay, male F344 rats � (x 21) 30 mg/kg/day 29

Long-term carcinogenesis assay, female F344 rats � 60 mg/kg/day 29

Long-term carcinogenesis assay, F344 rats � 1000 ppm in diet 30

26. Chlorpromazine (50-53-3)

DNA strand breaks, L5178Y cells in vitro � NT 2.5 mg/ml 31

DNA strand breaks, rat hepatocytes in vivo � 70 mg/kg po 32

Long-term carcinogenesis assay, male Syrian hamsters ? Dermal 33

27. Cimetidine (51481-61-9)

DNA strand breaks, transformed mouse eithelial cells in vitro � NT 1260 mg/ml 34

DNA strand breaks, rat primary hepatocytes + NT 756 mg/ml 34

UDS, rat primary hepatocytes + NT 83 mg/ml 34


UDS, rat primary hepatocytes + NT 25.2 mg/ml 34

DNA strand breaks, human primary hepatocytes � NT 2268 mg/ml 34


DNA strand breaks, rat hepatocytes in vivo � 250 mg/kg�1–20 d po 34

DNA strand breaks, rat gastric mucosa in vivo � 250 mg/ kg�1 po 34

Long term carcinogenesis assay, female C576BL/6 mice � (x 0.5) 226 mg/kg/day 34

Long-term carcinogenesis assay, CDR rats + (Leydig cell tumours) 378 mg/kg/day 5, 12

28. Ciprofibrate (52214-84-3)



Long-term carcinogenesis assay, male F344 rats + (liver tumours) 10 mg/kg in diet 12, 36

29. Citalopram (59729-32-7)

UDS, rat hepatocytes in vivo � NR 5

Long-term carcinogenesis assay, NMRI/BOM mice � (x 19) 240 mg/kg/day 5

Long-term carcinogenesis assay, COBS Wistar rats + (small intestine carcinomas) 8 mg/kg/day 5

30. Clobazam /22316-47-8)


DNA strand breaks, rat hepatocytes in vivo � 60 mg/kg/day po�15 days 10

Long-term carcinogenesis assay, rats � 300 mg/kg/day�9 months 37

Long-term carcinogenesis assay, rats + (thyroid tumours) 100 mg/kg/day 38

31. Clofibrate (637-07-0)

DNA strand breaks, L1210 cells in vitro � � 3 mg/ml 39


DNA strand breaks, rat hepatocytes in vivo � 200 mg/kg/day�6 days 39

DNA strand breaks, rat hepatocytes in vivo � 750 mg/kg in diet 39

DNA strans breaks, Fisher rat hepatocytes in vivo � 100 mg/kg in diet 39

UDS, rat hepatocytes in vivo � 750 mg/kg in diet�14 days 39

Binding (covalent) to DNA, male F344 rat hepatocytes in vitro � NT 243 mg/ml 39

Binding (covalent) to DNA, male F344 rat hepatocytes in vivo � 250 mg/kg/day�3 days 39

Long-term carcinogenesis assay, mice � (x 1.2) 5000 ppm in diet 39

Long term carcinogenesis assay, mice � (x 0.8) 350 mg/kg in diet 39

Long term carcinogenesis assay, F344 rats + (hepatocellular carcinomas) 2 50 mg/kg/day 39




Without e.m.s. With

e.m.s.

LED or HID

Long-term carcinogenesis assay, F344 rats + (hepatocellular carcinomas) 5000 ppm in diet 39

Long-term carcinogenesis assay, Sprague–Dawley rats + (liver nodules) 400 mg/kg/day 39

Long-term carcinogenesis assay, marmosets � 263 mg/kg/day 39

32. Clonazepam


DNA strand breaks, rat hepatocytes in vivo � 63 mg/kg/day�15 days 10

Long-term carcinogenesis assay, Wistar rat � 300 mg/kg/day 40

33. Clonidine (4205-90-7)

UDS, rat hepatocytes in vivo � 6 mg/kg 41



34. Clopidogrel (113665-84-2)

UDS, rat primary hepatocytes � NT NR 5, 7

Long-term carcinogenesis assay, mice � (x 4.9) 77 mg/kg/day 5, 7

Long-term carcinogenesis assay, rats � (x 10.1) 77 mg/kg/day 5, 7

35. Cloral hydrate (302-17-0)

DNA-protein cross-links, rat hepatocytes nuclei in vitro � NT 41250 mg/ml 42

DNA strand breaks, rat primary hepatocytes in vitro � NT 1650 mg/ml 42

DNA strand breaks, humam lymphoblastoid cells in vitro � NT 1650 mg/ml 42

DNA strand breaks, male Sprague–Dawley rat hepatocytes in vivo + 3000 mg/kg po�1 42

DNA strand breaks, male F344 rat hepatocytes in vivo � 1650 mg/kg po�1 42

DNA strand breaks, male B6C3F1 mouse hepatocytes in vivo + 100 mg/kg po 42

DNA strand breaks, male B6C3F1 mouse hepatocytes in vivo � 825 mg/kg po 42

Long-term carcinogenesis assay, male B6C3F1 mice + (liver tumours) 10 mg/kg po�1 42

Long-term carcinogenesis assay, male B6C3F1 mice + (liver tumours) 166 mg/kg/day po 42

Long-term carcinogenesis assay, male B6C3F1 mice + (liver tumours) 13.5 mg/kg/day po 42

Long-term carcinogenesis assay, female B6C3F1 mice + (lymphomas and

pituitary tumours)

100 mg/kg/day po 42

Long-term carcinogenesis assay, male B6C3F1 mice + (liver tumours) 25 mg/kg/day po 42

Long-term carcinogenesis assay, male B6C3F1 mice + (liver tumours) 0.165 mg ip in neonatal

animals

42

Long-term carcinogenesis assay, female B6C3F1 mice � 0.165 mg ip in neonatal

animals

42

Long-term carcinogenesis assay, Sprague–Dawley rats � (x 0.8) 135 mg/kg/day po 42

Long-term carcinogenesis assay, male F344 rats � (x 0.9) 162.6 mg/kg/day po 42

36. Clorazepate (5991-71-9)


DNA strand breaks, rat hepatocytes in vivo � 63.5 mg/kg/day po�15 d 10

Long-term carcinogenesis assay, dogs � 75 mg/kg/day 5

Long-term carcinogenesis assay, monkeys � 36 mg/kg/day 5

37. Cyclosporin (59865-13-3)

UDS, mouse sperm in vivo � NR 5, 34

Long-term carcinogenesis assay, mice + (lymphomas and liver tumours) 1–16 mg/kg/day 5

Long-term carcinogenesis assay, OF1 mice � (x 0.02) 16 mg/kg in diet 34

Long-term carcinogenesis assay, male AKR mice + (thymic lymphomas) 150 mg/kg in diet 34

Long-term carcinogenesis assay, rats + (pancreas and liver tumours) 0.5–8 mg/kg/day 5

Long-term carcinogenesis assay, OFA rats � (x 0.02) 8 mg/kg in diet 34

Long term carcinogenesis assay, beagle dogs � (x 4.3) 45 mg/kg/day 43

38. Cyproterone (2098-66-0)

DNA strand breaks, female rat primary hepatocytes + NT 20–50 mM 22, 44


UDS, female rat primary hepatocytes + NT 1–10 mM 22, 44




UDS, female rat primary hepatocytes + NT 3.16 mM 22, 45

UDS, female rat primary hepatocytes + NT 0.83 mM 22

DNA strand breaks, human males primary hepatocytes � NT 50 mM 22, 44

DNA strand breaks, human females primary hepatocytes + NT 50 mM 22, 44

UDS, human primary hepatocytes + NT 1–10 mM 22, 44

UDS, human males primary hepatocytes + NT 1–2 mM 22, 24

UDS, human females primary hepatocytes + NT 5 mM 22, 24

Binding (covalent) to male rat hepatocytes DNA in vitro (+) NT 3 mM 22, 23

Binding (covalent) to female rat hepatocytes DNA in vitro + NT 0. 3 mM 22, 23

Binding (covalent) to female rat hepatocytes in vivo + 1 mg/kg po 22, 23


Binding (covalent) to male rat hepatocytes in vivo + 100 mg/kg po 22, 47


Binding (covalent) to male C57BL/6 hepatocytes DNA in vivo � 35 mg/kg po 22

Long-term carcinogenesis assay, mice + (liver and stomach tumours) 96–104 mg/kg/day 12




Without e.m.s. With

e.m.s.

LED or HID

39. Danthron (117-10-2)

DNA strand breaks, L5178Ycells in vitro + NT 50 mM 48

UDS, mouse primary hepatocytes + NT 20 mM 49

UDS, rat primary hepatocytes + NT 20 mM 49

Long-term carcinogenesis assay, male C3H/HeN mice + (liver tumours) 0.2% in diet 34, 50

Long-term carcinogenesis assay, male rats + (large intestine tumours) 1% in diet 34

40. Diazepam (439-14-2)


DNA strand breaks, rat hepatocytes in vivo � 285 mg/kg po 10, 39

DNA strand breaks, rat hepatocytes in vivo � 57 mg/kg/day po�15 d 10, 39

Long-term carcinogenesis assay, CF-1 mice + (liver tumours) 75 mg/kg/day in diet 39

Long-term carcinogenesis assay, Wistar rats � (x 18.3) 75 mg/kg/day in diet 39

Long-term carcinogenesis assay, hamsters � 120 mg/kg/day in diet 39

Long-term carcinogenesis assay, gerbils � 10 mg/animal po weekly 39

41. Diphenhydramine (58-73-1)

DNA strand breaks, rat primary hepatocytes (+) NT 0.1 mM 51

UDS, rat primary hepatocytes � NT 0.1 mM 51


Long-term carcinogenesis assay, B6C3F1 mice � (x 2.9) 313 ppm in diet 7, 12, 27

Long-term carcinogenesis assay, F344 rats � 2000 ppm in diet 7, 12, 27

Long-term carcinogenesis assay, maleF344 rats ? (x 12.2) 635 ppm in diet 12, 27

Long-term carcinogenesis assay, female F344 rats ? (x 6.1) 313 ppm in diet 12, 27

42. Donepezil (120014-06-4)


Long-term carcinogenesis assay, CD-1 mice � 180 mg/kg/day 5

Long-term carcinogenesis assay, Sprague–Dawley rats � 30 mg/kg/day 5

43. Doxefazepam



Long-term carcinogenesis assay, Sprague–Dawley rats + (liver tumours) 30 mg/kg in diet 39, 52

44. Doxylamine (469-21-6)

UDS, rat primary hepatocytes (+) NT 0.5 mM 27, 53

Long-term carcinogenesis assay, B6C3F1 mice + (liver and thyroid tumours) 375–750 ppm in diet 12, 54

Long-term carcinogenesis assay, male F344 rats + (liver tumours) 115 mg/kg/day 12, 55

Long-term carcinogenesis assay, female F344 rats � 144 mg/kg/day 12, 55

45. Dronabinol (1972-08-3)

UDS, human fibroblast in vitro � NT NR 7

Long-term carcinogenesis assay, B6C3F1 mice ? 125 mg/kg/day 7, 56

Long-term carcinogenesis assay, F344 rats � (x 24) 50 mg/kg/day 7, 56

46. Duloxetine (116539-59-4)

UDS, rat primary hepatrocytes � NT NR 5, 6

Long-term carcinogenesis assay, male mice � (x 7.0) 100 mg/kg/day 5, 6

Long-term carcinogenesis assay, female mice + (liver tumours) 140 mg/kg/day 5, 6

Long-term carcinogenesis assay, rats � (x 5.1) 27–36 mg/kg/day 5, 6

47. Epinastine (80012-43-7)

UDS, rat hepatocytes in vivo � NR 5, 6



48. Eplerenone (107724-20-9)


Long-term carcinogenesis assay, p53 deficient mice � 1000 mg/kg/day 5

Long-term carcinogenesis assay, male rats + (thyroid tumours) 75 mg/kg/day 5

Long-term carcinogenesis assay, female rats + (thyroid tumours) 250 mg/kg/day 5

49. Estazolam (29975-16-4)


DNA strand breaks, rat hepatocytes in vivo � 57 mg/kg/day po�15 d 39

Long-term carcinogenesis assay, B6C3F1 mice � (x 24) 10 mg/kg in diet 5, 39, 57

Long-term carcinogenesis assay, Sprague–Dawley CD rats � 10 mg/kg in diet 5, 39, 57

50. Ethinylestradiol (57-63-6)

Binding (covalent) to female rat DNA of liver,

pancreas and kidney in vivo

+ 75 mg/day po�12 months 22

Binding (covalent to Syrian hamster kidney DNA in vivo + 22 mg impl�2 22

Long-term carcinogenesis assay, CF-LP mice + (pituitary tumours) 200–400 MRHD/kg/day 25

Long-term carcinogenesis assay, female Mead-Johnson rat � (x 16.9) 53 mg/kg/day 25

Long-term carcinogenesis assay, rats + (liver tumours) 200–400 MRHD/kg/day 25




Without e.m.s. With

e.m.s.

LED or HID

51. Fenofibrate (49562-28-9)



Long-term carcinogenesis assay, rats + (liver, pancreas and

testes tumours)

200 mg/kg/day 5

52. Finasteride (98319-26-7)

DNA strand breaks, animal cells in vitro � NT NR 5

Long-term carcinogenesis assay, male CD-1 mice 5, 12 + (Leydig cells tumours) 250 mg/kg/day 5, 12

Long-term carcinogenesis assay, male Sprague–Dawley rats � 160–320 mg/kg/day 5

53. Fluoxetine (54910-89-3)


Long-term carcinogenesis assay, B6C3F1 mice � (x 0.7) 12 mg/kg/day 7, 58

Long-term carcinogenesis assay, Sprague–Dawley rats � (x 1.2) 10 mg/kg/day 7, 58

54. Fluvastatin (93957-54-1)


Long-term carcinogenesis assay, male CD-1 mice � 350 mg/kg/day 7, 59

Long-term carcinogenesis assay, female CD-1 mice + (forestomach papillomas) 30 mg/kg/day 7, 59

Long-term carcinogenesis assay, CD-1 mice � 350 mg/kg/day 7, 59

Long-term carcinogenesis assay, mice + (gastric tumours) 15 mg/kg/day 6

Long-term carcinogenesis assay, CD rats � (x 5.8) 24 mg/kg/day 7, 59

Long-term carcinogenesis assay, male rats + (thyroid tumours) 24 mg/kg/day 7, 59

Long-term carcinogenesis assay, female rats � (x 5.8) 24 mg/kg/day 7, 59

55. Formoterol (75573-87-2)


UDS, human fibroblasts in vitro � � NR 5

Long-term carcinogenesis assay, mice + (adrenal tumours) 69 mg/kg/day in dr.wt 5

Long-term carcinogenesis assay, male mice + (hepatocarcinomas) 50 mg/kg/day in diet 5

Long-term carcinogenesis assay, female mice + (hepatocarcinomas) 20 mg/kg/day in diet 5

Long-term carcinogenesis assay, female mice + (uterine tumours) 2 mg/kg/day in diet 5

Long-term carcinogenesis assay, female rats + (ovarian tumours) 15 mg/kg/day in diet 5

Long-term carcinogenesis assay, female rats + (ovarian tumours) 0.5 mg/kg/day in diet 5

56. Frovatriptan (158747-02-05)



Long-term carcinogenesis assay, p53 transgenic mice ? 400 mg/kg/day 6

Long-term carcinogenesis assay, male rats + (pituitary tumours) 85 mg/kg/day 6

Long-term carcinogenesis assay, female rast � 85 mg/kg/day 6

57. Gabapentin (60142-96-3)


Long-term carcinogenesis assay, mice � (x 8.0) 2000 mg/kg/day 5

Long-term carcinogenesis assay, male Wistar rats + (pancreas tumours) 2000 mg/kg/day 5, 60

Long-term carcinogenesis assay, female Wistar rats � (x 16.2) 2000 mg/kg/day 5, 60

58. Glibenclamide (10238-21-8)



59. Glimepiride (93479-97-1)


Long-term carcinogenesis assay, mice + (pancreas adenomas) >320 ppm in diet 5

Long-term carcinogenesis assay, rats � 5000 ppm in diet 5

60. Hydralazine (86-54-4)

DNA strand breaks, Chinese hamster lung V79 cells in vitro + NT 90 mg/ml 61




UDS, human primary hepatocytes (+) NT 160 mg/ml 61

DNA strand breaks, Swiss mice liver in vivo + 67.3 mg/kg 63

DNA strand breaks, Swiss mice lung in vivo � 67.3 mg/kg 63

DNA strand breaks, Swiss mice liver, kidney, spleen in vivo + 83 mg/kg 64

DNA strand breaks, Swiss mice lung in vivo � 83 mg/kg 64

UDS, RAIf rat liver in vivo � 125 mg/kg 65

UDS, RAIf rat liver in vivo � 60 mg/kg/day�7 days 65

Long-term carcinogenesis assay, Swiss mice + (lung tumours) 0.125% in dr.wt. 66

Long-term carcinogenesis assay, Sprague–Dawley rats � 0.035% in diet 67

Long-term carcinogenesis assy, male rats + (testes tumours) 60 mg/kg/day 68

Long-term carcinogenesis assay, female rats + (liver nodules) 30 mg/kg/day 68

61. Irbesartan (138402-11-6)


Long-term carcinogenesis assay, mice � (x 16) 1000 mg/kg/day 5, 6




Without e.m.s. With

e.m.s.

LED or HID

Long-term carcinogenesis assay, male rats � (x 16.2) 500 mg/kg/day 5, 6

Long-term carcinogenesis assay, female rats � 1000 mg/kg/day 5, 6

62. Isotretinoin (4759-48-2)


Long-term carcinogenesis assay, F344 rats + (pheochromocytomas) 8–32 mg/kg/day 5

63. Isradipine (75695-93-1)


Long-term carcinogenesis assay, male CD-1 mice ? 80 mg/kg/day 5, 6

Long-term carcinogenesis assay, female CD-1 mice � 80 mg/kg/day 5, 6

Long-term carcinogenesis assay, male F344 rats + (testes tumours) 12.5 mg/kg/day 5, 6

Long-term carcinogenesis assay, female F344 rats � 62.5 mg/kg/day 5, 6

64. Lansoprazole (103577-45-3)


Long-term carcinogenesis assay, male CD-1 mice + (liver and testes tumours) 75–300 mg/kg/day 5, 7

Long-term carcinogenesis assay, female CD-1 mice + (liver tumours) 150 mg/kg/day 5

Long-term carcinogenesis assay, male Sprague–Dawley rats + (stomach and testes tumours) 15–150 mg/kg/day 5

Long-term carcinogenesis assay, femaleSprague –Dawley rats + (stomach tumours) 15–150 mg/kg/day 5

Long-term carcinogenesis assay, Wistar rats + (stomach tumours) NR 5

65. Leflunomide (75706-12-6)


Long-term carcinogenesis assay, mice + (lymphomas) 15 mg/kg/day 5, 7


66. Lisinopril (76547-98-3)

DNA strand breaks, rat primary hepatocytes � NT NR 5, 6



67. Loratadine (79794-75-5)


Long-term carcinogenesis assay, male mice + (liver tumours) 40 mg/kg/day 7

Long-term carcinogenesis assay, female mice � (x 19.2) 40 mg/kg/day 7

Long-term carcinogenesis assay, male rats + (liver tumours) 10 mg/kg/day 7

Long-term carcinogenesis assay, female rats + (liver tumours) 25 mg/kg/day 7

68. Lorazepam (846-49-1)



Long-term carcinogenesis assay, CD rats � 0.48% in diet 69

69. Losartan (114798-26-4)

DNA strand breaks, animal cells in vitro � NT NR 5, 6


Long-term carcinogenesis assay, male rats � 270 mg/kg/day 5, 6

Long-term carcinogenesis assay, female rats ? 270 mg/kg/day 5, 6

70. Lovastatin (75330-75-5)

DNA strand breaks, mouse primary hepatocytes � NT 121 mg/ml 7


Long-term carcinogenesis assay, mice + (liver, lung and stomach tumours) 500 mg/kg/day 7, 70

Long-term carcinogenesis assay, rats + (liver and thyroid tumours) 30 mg/kg/day 7, 70

71. Mepyramine

UDS, rat primary hepatocytes + NT 10 mM 27, 53


Long-term carcinogenesis assay, B6C3F1 mice � (x 9.6) 1500 ppm in diet 71

Long-term carcinogenesis assay, female Sprague–Dawley rats � (x 13) 1000 ppm in dr.wt. 7, 72

Long-term carcinogenesis assay, F344 rats � 3000 ppm in diet 73

Long-etrm carcinogenesis assay, F344 rats � 2000 ppm in dr.wt. 74

Long-term carcinogenesis assay, F344 rat � 2000 ppm in diet 74

Long-term carcinogenesis assay, rats + (liver tumours) NR 27

72. Methapyrilene (91-80-5)

DNA strand breaks, rat primary hepatocytes + NT 0.5 mM 75

UDS, rat primary hepatocytes � NT 20 mM 53



UDS, rat primary hepatocytes � + 1 mM 28

DNA (covalent binding, rat liver in vivo � 2 g/kg 77

DNA (covalent) binding in vitro � + 500 mg/ml 27, 78

DNA (covalent) binding in vitro NT � 500 mg/ml 77, 79

DNA (covalent) binding E.coli in vitro ? NT 30 mM 78

Long-term carcinogenesis assay, F344 rats + (liver tumours) 125 ppm in diet 80




Without e.m.s. With

e.m.s.

LED or HID

73. Metoprolol (54163-88-1)






DNA strand breaks, Swiss mice liver and kidney in vivo � 1150 mg/kg 81

DNA strand breaks, Sprague–Dawley rats,liver

and gastric mucosa in vivo

� 1000 mg/kg 81

Long-term carcinogenesis assay, male Swiss mice � (x 9.0) 750 mg/kg/day 5, 6

Long-term carcinogenesis assay, female Swiss mice + (lung adenomas) 750 mg/kg/day 5, 6

Long-term carcinogenesis assay, CD-1 mice � (x 9.0) 750 mg/kg/day 5, 6


74. Mibefradil (116644-53-2)



Long-term carcinogenesis assay, rats ? (x 3.4) 35 mg/kg/day 6

75. Midazolam (59467-70-8)




Long-term carcinogenesis assay, rats + (thyroid tumours) 80 mg/kg/day 7

76. Minoxidil (38304-91-5)



Long-term carcinogenesis assay, male mice + (liver nodules) 63 mg/kg/day po 6

Long-term carcinogenesis assay, female mice + (lymphomas) 10 mg/kg/day po 6

Long-term carcinogenesis assay, male mice � 80 mg/kg/day (skin) 6

Long-term carcinogenesis assay, female mice + (mammary tumours) 8 mg/kg/day (skin) 6


Long-term carcinogenesis assay, rats + (pheochromocytomas and

preputial gland tumours)

NR (dermal study) 6

77. Mirtazapine (61337-67-5)

UDS, HeLa cells in vitro � NT NR 6


Long-term carcinogenesis assay, female mice � (x 21) 200 mg/kg/day 6

Long-term carcinogenesis assay, male rats + (liver and thyroid tumours) 60 mg/kg/day 6

Long term carcinogenesis assay, female rats + (liver tumours) 20 mg/kg/day 6

78. Modafinil (68693-11-8)


Long-term carcinogenesis assay, mice ? (x 1.4) 60 mg/kg/day 5


79. Montelukast (158966-92-8)

DNA strand breaks, rat primary hepatocytes � NT NR 5



80. Morphine (57-27-2)

DNA strand breaks, B6C3F1 mice thymocytes in vitro � NT 28.5 mg/ml 82

DNA strand breaks, human T cells in vitro + NT NR 5

DNA strand breaks, B6C3F1 mice thymocytes in vivo + 75 mg/animal 82

Long-term carcinogenesis assay, C57BL/6 mice � (x 0.07) 10 mg/kg/week po 83

Long-term carcinogenesis assay, CBA mice ? (x 0.6) 0.3 mg/animal sc 83

Long-term carcinogenesis assay, Swiss mice � (x 0.04) 0.06 mg/animal (skin) 83

Long-term carcinogenesis assay, Syrian golden hamsters ? 0.284 mg/animal

intratracheal

83

81. Nadolol (42200-33-9)






Long-term carcinogenesis assay, CD-1 mice � (x 3.7) 500 mg/kg/day 84

Long-term carcinogenesis assay, CD rats �(x 15.2) 1000 mg/kg/day 84

82. Naltrexone (16676-29-2)

-UDS, human diploid fibroblast in vitro + NT NR 7

-Long-term carcinogenesis assay, rats + (mesotheliomas and

vascular tumours)

NR 6

83. Nefazodone (83366-66-9)





Without e.m.s. With

e.m.s.

LED or HID


Long-term carcinogenesis assay,rats � (x 3.2) 200 mg/kg/day 6

84. Norethisterone (68-22-4)

UDS, female rat primary hepatocytes � NT 150 mg/ml 22

UDS, male rat primary hepatocytes (+) NT 15 mg/ml 22

Long-term carcinogenesis assay, male CF-LP mice + (liver tumours) 200–400 MRHD/kg/day 25

Long-term carcinogenesis assay, female CF-LP mice + (pituitary tumours) 200–400 MRHD/kg/day 25

Long-term carcinogenesis assay, male rats + (liver and mammary tumours) 200–400 MRHD/kg/day 25

85. Olanzapine (132539-06-1)


Long-term carcinogenesis assay, male mice � (x 5.6) 20 mg/kg/day 5

Long-term carcinogenesis assay, female mice + (liver tumours) 3 mg/kg/day 5

Long-term carcinogenesis assay, male mice � (x 2.2) 8 mg/kg/day 5

Long-term carcinogenesis assay, female mice + (mammary tumours) >2 mg/kg/day 5

Long-term carcinogenesis assay, male rats � (x 2.3) 4 mg/kg/day 5

Long-term carcinogenesis assay, female rats + (mammary tumours) 8 mg/kg/day 5

86. Olmesartan (144689-24-7)

DNA strand breaks, rat kidney in vivo � NR 6, 68

Long-term carcinogenesis assay, p53 knockout mice � 1000 mg/kg/day 5, 6

Long-term carcinogenesis assay, Hras 2 transgenic mice � 1000 mg/kg/day 5, 6


87. Omeprazole (73590-58-6)

UDS, rat primary hepatocytes (+) NT 10 mg/ml 7, 85

UDS, human primary hepatocytes (+) NT 10 mg/ml 7, 85

DNA strand breaks, Sprague–Dawley rats liver


� 100 mg/kg�3 d. 86

Long-term carcinogenesis assay, CD-1 mice ? NR 5, 12

Long-term carcinogenesis assay, p53 transgenic mice � NR 5

Long-term carcinogenesis assay, rats + (gastric carcinoids) 1.7–140.8 mg/kg/day 5

Long-term carcinogenesis assay, female rats � (x 1.1) 13.8 mg/kg/day 5

Long-term carcinogenesis assay, Sprague–Dawley rats � (x 11.4) 140.8 mg/kg/day 5, 12

Long-term carcinogenesis assay, male Sprague–Dawley rats (+) 0.4–16 mg/kg/day 5

Long-term carcinogenesis assay, female Sprague–Dawley rats � (x 1.3) 0.4–16 mg/kg/day 5

88. Oxaprozin (21256-18-8)

UDS, Chinese hamster ovary cells in vitro � NT NR 6

Long-term carcinogenesis assay, male CD-1 mice + (liver tumours) NR 6

Long-term carcinogenesis assay, female CD-1 mice � NR 6

Long-term carcinogenesis assay, rats � NR 6

89. Oxazepam (604-75-1)

DNA strand breaks, human myeloid leukaemia K562 cells + NT 287 mg/ml 87

UDS, rat primary hepatocytes � NT 0.5 mg/ml 39

DNA strand breaks, rat liver in vivo � 287 mg/kg po 10, 39

DNA strand breaks, rat liver in vivo � 57 mg/kg/day po�15 d. 10, 39

Long-term carcinogenesis assay, Swiss mice + (liver tumours) 2500 ppm in diet 39, 88, 89

Long-term carcinogenesis assay, B6C3F1 mice + (liver tumours) 2500 ppm in diet 88, 89

Long-term carcinogenesis assay, F344/N rats ? (renal adenomas) 2500 ppm in diet 90, 91

90. Oxprenolol (6452-71-7)









� 107 mg/kg 13

Long-term carcinogenesis assay, CF-1 mice � (x 1.5) 150 mg/kg/day 92


91. Pantoprazole (138786-67-1)


Binding (covalent) to rat liver DNA in vivo ? NR 5, 7

Long-term carcinogenesis assay, p53 transgenic mice � NR 5, 7

Long-term carcinogenesis assay, male B6C3F1 mice � (x 18) 150 mg/kg/day 5, 7

Long-term carcinogenesis assay, female B6C3F1mice + (liver tumours) 150 mg/kg/day 5, 7

Long-term carcinogenesis assay, Sprague–Dawley rats + (stomach, liver and

thyroid tumours)

50–200 mg/kg/day 5, 7

Long-term carcinogenesis assay, F344 rats + (gastric tumours) 5–50 mg/kg/day 7

92. Paracetamol (103-90-2)

DNA strand breaks, Reuber H4-II-E rat hepatoma cells in vitro � NT 1510 mg/ml 93

DNA strand breaks, Chinese hamster lung V79 cells in vitro (+) NT 1510 mg/ml 93




Without e.m.s. With

e.m.s.

LED or HID

UDS, rat primary hepatocytes � NT 1 mM 11

DNA strand breaks, mouse liver cells in vivo + 600 mg/kg ip�1 93

DNA strand breaks, mouse kidney cells in vivo � 600 mg/kg ip�1 93

DNA strand breaks, rat liver and kidney cells in vivo � 600 mg/kg ip�1 93

Binding (covalent) to ICR mouse liver DNA in vivo + 300 mg/kg�1 ip 93

Long-term carcinogenesis assay, IF mice + (liver tumours) 1000 mg/kg of diet 93

Long-term carcinogenesis assay, B6C3F1 mice � (x 1.0) 6000 mg/kg of diet 93

Long-term carcinogenesis assay, B6C3F1 mice � (x 1.6) 10,000 mg/kg of diet 34

Long-term carcinogenesis assay, Sprague–Dawley rats ? 5300 mg/kg of diet 34

Long-term carcinogenesis assay, F344 rats � (x 4.2) 13000 mg/kg of diet 34

Long-term carcinogenesis assay, Leeds rats + (liver and bladder tumours) 5000/10,000 mg/kg of diet 34

Long-term carcinogenesis assay, male F344 rats � (x 2.0) 6000 mg/kg of diet 93

Long-term carcinogenesis assay, female F344 rats + (leukaemias) 6000 mg/kg of diet 93

93. Paroxetine 61869-08-7)

UDS, animal cells in vitro _ NT NR 5

Long-term carcinogenesis assay, mice ? (x 2.4) 25 mg/kg/day 5

Long-term carcinogenesis assay, male rats + (reticulum cell sarcomas) 20 mg/kg/day 5


94. Penbutolol (38363-40-5)

DNA strand breaks, human fibroblasts in vitro + NT 40 mg/ml 94

DNA strand breaks, A549 human lung tumour cells in vitro � � 30 mg/ml 95

UDS, A549 human lung tumour cells in vitro � � 30 mg/ml 95

DNA strand breaks, Wistar rats liver in vivo � 100 mg/kg 95



95. Pentoxyfylline (6493-05-6)

UDS, animal cells in vitro � � NR 5



96. Pergolide (66104-22-1)


Long-term carcinogenesis assay, male mice � 36.4 mg/kg/day 5

Long-term carcinogenesis assay, female mice + (uterine tumours) 40.8 mg/kg/day 5

Long-term carcinogenesis assay, male rats � (x 2.9) 0.88 mg/kg/day 5

Long-term carcinogenesis assay, female rats + (uterine tumours) 1.42 mg/kg/day 5

97. Phenazopyridine (94-78-0)

UDS, rat primary hepatocytes + NT 0.1 mM 96

DNA strand breaks, ddy male mice stomach, colon, liver in vivo � 400 mg/kg po 97

DNA strand breaks, ddy male mice kidney, bladder, lung,

brain and bone-marrow in vivo

� 400 mg/kg po 97

Long-term carcinogenesis assay, B6C3F1 mice + 600 mg/kg in diet 98

Long-term carcinogenesis assay, A/He mice � 1550 mg/kg weekly ip 98

Long-term carcinogenesis assay, femaleF344 rats + (colon and rectum tumours) 3700 mg/kg in diet 98

98. Phenelzine (51-71-8)

DNA strand breaks, Swiss mice liver in vivo + 312 mg/kg po 99, 100

DNA strand breaks, Swiss mice lung in vivo + 56 mg/kg�3 ip 99, 100

Long-term carcinogenesis assay, mice + (lung and blood

vessels tumours)

0.015% in dr.wt. 98

Long-term carcinogenesis assay, Sprague–Dawley rats � (x 2.5) 0.02% in diet 67

99. Phenobarbital (50-06-6)

DNA strand breaks, Chinese hamster lung V79 cells in vitro � � 2320 mg/ml 101



DNA strand breaks, Chinese hamster ovary cells in vitro � � 1160 mg/ml 101



DNA strand breaks, male CD-1 mice liver in vivo + 140 mg/kg 101

DNA strand breaks, male CD-1 mice lung, spleen, kidney

and bone-marrow in vivo

� 140 mg/kg 101

Binding (covalent) to B6C3F1 mouse liver DNA in vivo � 200 mg/kg po�1 101

Binding (covalent) to B6C3F1 mouse liver DNA in vivo � 1000 ppm in diet�2 wk 101

Long-term carcinogenesis assay, C3H mice + (liver tumours) 0.05% in diet 101

Long-term carcinogenesis assay, CF1 mice + (liver tumours) 0.05% in diet 101

Long-term carcinogenesis assay, CF1 mice + (liver tumours) 0.05% in dr.wt. 101

Long-term carcinogenesis assay, BALB/c mice � (x 3.7) 0 .05% in diet 101

Long-term carcinogenesis assay, male Avy/A mice + (liver tumours) 0.05% in diet 101

Long-term carcinogenesis assay, C3H mice + (liver tumours) 0.02% in diet 101

Long-term carcinogenesis assay, C3H mice ? (x 3.1) 85 mg/kg/day 101

Long-term carcinogenesis assay, C57BL/6 mice ? (x 3.1) 85 mg/kg/day 101

Long-term carcinogenesis assay, C-myc mice + (liver tumours) 0.05% in diet 101




Without e.m.s. With

e.m.s.

LED or HID

Long-term carcinogenesis assay, C57BL/6 p53 heterozygous mice � (x 3.7) 0.1% in diet 101

Long-term carcinogenesis assay, MT42 transgenic mice + (liver tumours) 0.05% in diet 101

Long-term carcinogenesis assay, Wistar rats + (liver tumours) 0.05% in dr.wt. 101

Long-term carcinogenesis assay, F344 rats ? (x 3.8) 0.05% in diet 101

Long-term carcinogenesis assay, F344 rats ? (x 7.5) 0.05% in dr.wt. 101

100. Phenolphthalein (77-09-8)

DNA strand breaks, blood leukocytes of transgenic ? 2074 mg/kg in diet 102

TSG-p53 mice in vivo x 6 months

Binding (covalent) to Syrian hamster embryo cells DNA in vitro � NT 12.8 mg/ml 102

Long-term carcinogenesis assay, p53 (+/�) mice + (lymphomas) 3000 mg/kg in diet 102

Long-term carcinogenesis assay, B6C3F1 mice + (lymphomas and sarcomas) 3000–6000 mg/kg in diet 102

Long-term carcinogenesis assay, F344 rats + (pheochromocytomas

and lymphomas)

12,500 mg/kg in diet 102

101. Pilocarpine (92-13-7)



Long-term carcinogenesis assay, Sprague–Dawley rats + (pheochromocytomas

and liver adenomas)

18 mg/kg/day 5

102. Pindolol (13523-86-9)


DNA strand breaks, Sprague–Dawley rats liver and

gastric mucosa in vivo

� 132 mg/kg 81



103. Pioglitazone (111025-46-8)

UDS, rat primary hepatocytes � NT NR 5, 6, 7

Long-term carcinogenesis assay mice � (x 12.4) 100 mg/kg/day 5, 6, 7

Long-term carcinogenesis assay, male rats + (urinary bladder tumours) 4 mg/kg/day 5, 6, 7

Long-term carcinogenesis assay, female rats � (x 15.9) 63 mg/kg/day 5, 6, 7

104. Potassium canrenoate (4138-96-9)



DNA strand breaks, human lymphocytes in vitro � NT 36 mg/ml 103

DNA strand breaks, primary hepatocytes + NT 4 mg/ml 103

UDS, human primary hepatocytes + NT 4 mg/ml 103

DNA strand breaks, male Sprague–Dawley rat liver in vivo (+) 325 mg/kg 104

DNA strand breaks, male Sprague–Dawley rat

thyroid and bone-marrow in vivo

� 325 mg/kg 104

DNA strand breaks, female Sprague–Dawley rat liver in vivo � 325 mg/kg 104

DNA strand breaks, female Sprague–Dawley rat

thyroid and bone-marrow in vivo

(+) 325 mg/kg 104

DNA strand breaks, Sprague–Dawley rats testes

and ovary in vivo

+ 81 mg/kg 104

UDS, rat liver in vivo � 325 mg/kg 104

Long-term carcinogenesis assay, Sprague–Dawley rats + (myelocytic leukaemia) 20 mg/kg 6

105. Prazepam (2955-38-6)

DNA strand breaks, rat liver in vivo � 325 mg/kg po 10, 39

DNA strand breaks, rat liver in vivo � 65 mg/kg/day po�15 d 10, 39

Long-term carcinogenesis assay, CF-1 mice � (x 6.0) 75 mg/kg/day in diet 39

Long-term carcinogenesis assay, Wistar rats � (x 6.0) 75 mg/kg/day in diet 39

106. Promethazine (60-87-7)

DNA strand breaks, rat primary hepatocytes � NT 0.1 mM 105


UDS, rat liver in vivo � 400 mg/kg 106

Long-term carcinogenesis assay, male B6C3F1 mice � (x 2.2) 45 mg/kg/day 107

Long-term carcinogenesis assay, female B6C3F1 mice � (x 0.7) 15 mg/kg/day 107

Long-term carcinogenesis assay, F344/N rats � (x 3.2) 33.3 mg/kg/day 107

107. Propranolol (525-66-6)





DNA strand breaks, human fibroblasts in vitro � NT 65 mg/ml 94



DNA strand breaks, rat liver in vivo � 75 mg/kg 13



and gastrixc mucosa in vivo

� 233 mg/kg 81




Without e.m.s. With

e.m.s.

LED or HID



Long-term carcinogenesis assay, Long-Evans rats � (x 0.6) 37.5 mg/kg/day 108


108. Propylthiouracil (51-52-5)

DNA strand breaks, rat primary hepatocytes � NT 953 mg/m 101

DNA strand breaks, human primary hepatocytes � NT 953 mg//ml 101

Long-term carcinogenesis assay, male A mice + (thyroid tumours) 0.8% in diet 101

Long-term carcinogenesis assay, C57BL mice + (pituitary adenomas) 10 g/kg in diet 101

Long-term carcinogenesis assay, rats + (thyroid tumours) 0.1% in dr.wt. 101

Long-term carcinogenesis assay, Wistar rats + (thyroid tumours) 0.025–0.2% in dr.wt. 101

Long-term carcinogenesis assay, Long Evans rats + (thyroid tumours) 0.1% in diet 101

Long-term carcinogenesis assay, rats + (thyroid tumours) 0.006% in dr.wt. 101

Long-term carcinogenesis assay, Syrian golden hamster + (thyroid tumours) 0.2% in dr.wt. 101

Long-term carcinogenesis assay, male guinea-pigs + (thyroid tumours) 0.03% in dr.wt. 101

109. Quazepam (36735-22-5)

DNA strand breaks, rat liver in vivo � 325 mg/kg po 10

DNA strand breaks, rat liver in vivo � 65 mg/kg/day po�15 d 10

Long-term carcinogenesis assay, CF-1 mice � (x 6.0) 75 mg/kg/day in diet 109

Long-term carcinogenesis assay, Wistar rats � (x 12) 75 mg/kg/day in diet 109

110. Rabeprazole (117976-89-3)

UDS, rat liver in vivo � NR 5, 7

Long-term carcinogenesis assay, CD-1 mice � (x 4.7) 100 mg/kg/day 5, 7

Long-term carcinogenesis assay, male Sprague–Dawley rats � (x 5.7) 60 mg/kg/day 5, 7

Long-term carcinogenesis assay, female Sprague–Dawley rats + (gastric tumours) 5 mg/kg/day 5, 7

111. Raloxifene (84449-90-1)


Long-term carcinogenesis assay, male mice + (testes and prostate tumours) 41–210 mg/kg/day 5

Long-term carcinogenesis assay, female mice + (ovarian tumours) 9–242 mg/kg/day 5

Long-term carcinogenesis assay, rats + (ovarian tumours) 279 mg/kg/day 5

112. Ramipril (87333-19-5)

UDS, human cells in vitro � � NR 5, 6



113. Ranitidine (66357-35-5)

DNA strand breaks, Chinese hamster ovary

cells (CHO-K1) cells in vitro

� NT 3 mM 110


DNA strand breaks, rat hepatocytes in vivo � 350 mg/kg/day�5 d 112

DNA strand breaks, rat gastric mucosa in vivo � 350 mg/kg po 112

Long-term carcinogenesis assay, mice � (x 14) 2000 mg/kg/day 5, 7

Long-term carcinogenesis assay, rats � (x 28) 2000 mg/kg/day 5, 7

114. Rasagiline (136236-51-6)

UDS, animal cells in vivo � NR 6

Long-term carcinogenesis assay, CD-1 mice + (lung tumours) 15 mg/kg/day 6

vLong-term carcinogenesis assay, Sprague–Dawley rats � 3–17 mg/kg/day 6

115. Repaglinide (135062-02-1)



Long-term carcinogenesis assay, male rats + (liver and thyroid tumours) 30–60 mg/kg/day 5

Long-term carcinogenesis assay, female rats � 120 mg/kg/day 5

116. Reserpine (50-55-5)

UDS, rat primary hepatocytes � NT 6.1 mg/ml 113


UDS, Fisher rat liver in vivo � 400 mg/kg 114

Binding (covalent) to Syrian hamster embryo cells DNA in vitro � 8 mg/ml 115

Long-term carcinogenesis assay, male B6C3F1 mice + (carcinomas of

seminal vesicles)

10 ppm in diet 116

Long-term carcinogenesis assay, female B6C3F1 mice + (mammary tumours) 5 ppm in diet 116

Long-term carcinogenesis assay, male F344 rats + (pheochromocytomas) 5 ppm in diet 116

Long-term carcinogenesis assay, female F 344 rats � 10 ppm in diet 116

117. Rifampicin (13292-46-1)

Binding (covalent) to DNA in vitro � NT NR 98

Long-term carcinogenesis assay, male C3Hf mice � (x 0.5) 0.06% in dr.wt 7, 98

Long-term carcinogenesis assay, female C3Hf mice + (liver tumours) 0.03% in dr.wt. 98

Long-term carcinogenesis assay, BALB/c mice � (x 0.5) 0.06% in dr.wt 98

Long-term carcinogenesis assay, BALB/c mice � (x 0..05) 0.3 mg/mouse sc 98

Long-term carcinogenesis assay, Wistar rats � (x 1.0) 0.06% in dr.wt. 98




Without e.m.s. With

e.m.s.

LED or HID

118. Risedronate (105462-24-6)




119. Risperidone (106266-06-2)


Long-term carcinogenesis assay, male Swiss mice � (x 3.5) 10 mg/kg/day 5

Long-term carcinogenesis assay, female Swiss mice + (pituitary and

mammary tumours)

0.63 mg/kg/day 5

Long-term carcinogenesis assay, male Wistar rats + (pancreas and

mammary tumours)

2.5 mg/kg/day 5

Long-term carcinogenesis assay, female Wistar rats + (mammary tumours) 0.63 mg/kg/day 5

120. Rivastigmine (123441-03-2)


Long-term carcinogenesis assay, mice � (x 0.6) 1.6 mg/kg/day 5


121. Rizatriptan (144034-80-0)




122. Rofecoxib (162011-90-7)

DNA strand breaks, animal cells in vitro � NT NR 5, 7

DNA strand breaks, animal cells in vivo � NR 5, 7

Long-term carcinogenesis assay, mice � (x 5.8) 30–60 mg/kg/day 5, 7


123. Rosiglitazone (122320-73-4)



Long-term carcinogenesis assay, Sprague–Dawley rats + (lipomas) 30.3 mg/kg/day 5

124. Sibutramine (106650-56-0)

UDS, HeLa cells in vitro � � NR 5



Long-term carcinogenesis assay, male rats + (testes tumours) 9 mg/kg/day 5

Long-term carcinogenesis assay, female rats � (x 5.8) 9 mg/kg/day 5

125. Simvastatin (79902-63-9)


Long-term carcinogenesis assay, mice + (liver and lung tumours) 100 mg/kg/day 5

Long-term carcinogenesis assay, female rats + (thyroid adenomas) 25 mg/kg/day 5


Long-term carcinogenesis assay, rats + (liver and thyroid tumours) 50 mg/kg/day 5

126. Sotalol (3930-20-9)







DNA strand breaks, Sprague–Dawley rats

liver and gastric mucosa in vivo

� 1725 mg/kg 81

Long-term carcinogenesis assay, CD-1 mice � 600 mg/kg/day 108


Long-term carcinogenesis assay, Long Evans rats � (x 8.4) 275 mg/kg/day 108

127. Tacrolimus

UDS, rodents primary hepatocytes � NT NR 5

Long-term carcinogenesis assay, mice � (x 0.2) 0.5 mg/kg/day 5


128. Tamsulosin


Long-term carcinogenesis assay, male B6C3F1 mice � 127 mg/kg/day 6

Long-term carcinogenesis assay, female B6C3F1 mice + (mammary tumours) 45 mg/kg/day 5, 6

Long-term carcinogenesis assay, male F344 rats � 43 mg/kg/day 5, 6

Long-term carcinogenesis assay, female F344 rats + (mammary tumours) 5.4 mg/kg/day 5, 6

129. Tegaserod (145158-71-0)


Long-term carcinogenesis assay, mice + (small intestine

adenocarcinomas)

600 mg/kg/day 5, 6





Without e.m.s. With

e.m.s.

LED or HID

130. Temazepam (846-50-4)



Long-term carcinogenesis assay, CD-1 mice ? (x 19.2) 1600 mg/kg in diet 39, 117

Long-term carcinogenesis assay, CD rats � 1600 mg/kg in diet 39, 117

131. Terbinafine


Long-term carcinogenesis assay, male rats + (liver tumours) 69 mg/kg/day 5

132. Thenyldiamine (91-79-2)

UDS, rat primary hepatocytes (+) NT 1 mM 53

Long-term carcinogenesis assay, Sprague–Dawley rats � 1000 ppm in diet 73

Long-term carcinogenesis assay, male F344 rats � 17.6 mg/kg/day 12

Long-term carcinogenesis assay, female F344 rats � 25.1 mg/kg/day 12

133. Tiagabine (115103-54-3)



Long-term carcinogenesis assay, rats + (liver and Leydig cell

tumours)

200 mg/kg/day 5

134. Timolol (26839-75-8)

DNA strand breaks, human fibroblasts in vitro � NT 125 mg/ml 94

Long-term carcinogenesis assay, male mice � 500 mg/kg/day 5, 6

Long-term carcinogenesis assay, female mice + (lung, uterus and

mammary tumours)

500 mg/kg/day 5, 6

Long-term carcinogenesis assay, male rats + (pheochromocytomas) 300 mg/kg/day 5, 6

Long-term carcinogenesis assay, female rats � 300 mg/kg/day 5, 6

135. Tiotropium (186691-13-4)


Long-term carcinogenesis assay, male mice � (x 0.5) 0.02 mg/kg/day 5

Long-term carcinogenesis assay, female mice � (x 3.6) 0.145 mg/kg/day 5

Long-term carcinogenesis assay, rats � 0.059 mg/kg/day 5

136. Tizanidine (51322-75-9)

UDS, mouse cells in vivo � NR 6



137. Tolcapone (134308-13-7)



Long-term carcinogenesis assay, rats + (uterus and kidney

tumours)

250 mg/kg/day 5

138. Topiramate (97240-79-4)


Long-term carcinogenesis assay, mice + (urinary bladder tumours) 300 mg/kg/day 5


139. Torasemide (56211-40-6)


UDS, mouse primary hepatocytes � NT NR 5


Long-term carcinogenesis assay, male rats � 9 mg/kg/day 5

Long-term carcinogenesis assay, female rats (+) (kidney tumours) 9 mg/kg/day 5

140. Triazolam (28911-01-5)


DNA strand breaks, rat hepatocytes in vivo � 69 mg/kg/day�15 d 10

Long-term carcinogenesis assay, mice � 4000MRHD/kg/day 7

141. Troglitazone


Long-term carcinogenesis assay, male mice + (hemangiosarcomas) 800 mg/kg/day 6

Long-term carcinogenesis assay, female mice + (hepatocarcinomas and

hemangiosarcomas)

400 mg/kg/day 6

Long-term carcinogenesis assay, male rats � 800 mg/kg/day 6

Long-term carcinogenesis assay, female rats � (x 12.0) 200 mg/kg/day 6

142. Zaleplon (151319-34-5)


Long-term carcinogenesis assay, mice + (liver tumours) 200 mg/kg/day in diet 5

Long-term carcinogenesis assay, mice � NR 118

Long-term carcinogenesis assay, rats � NR 118

Long-term carcinogenesis assay, rats � (x 9.8) 20 mg/kg/day in diet 5




Without e.m.s. With

e.m.s.

LED or HID

143. Zileuton (111406-87-2)


UDS, mouse kidney cells in vitro � NT NR 6

UDS, rat kidney cells in vitro � NT NR 6


Long-term carcinogenesis assay, female mice + (liver, kidney and

vessels tumours)

450 mg/kg/day 6

Long-term carcinogenesis assay, rats + (kidney tumours) 170 mg/kg/day 6

144. Zolmitriptan (139264-17-8)

UDS, mammalian cells uin vitro � NT NR 5


Long-term carcinogenesis assay, male rats + (thyroid tumours) 400 mg/kg/day 5

Long-term carcinogenesis assay, female rats � 400 mg/kg/day 5

145. Zolpidem (82626-48-0)


Long-term carcinogenesis assay, mice � 80 mg/kg/day in diet 5

Long-term carcinogenesis assay, rats � 80 mg/kg/day in diet 5

146. Zopiclone (43200-80-2)


Long-term carcinogenesis assay, B6C3F1 mice + (lung and skin tumours) 100 mg/kg/day po 6

Long-term carcinogenesis assay, Sprague–Dawley rats + (mammary and thyroid

tumours)

100 mg/kg/day po 6

Note. The name of each drug is followed by the corresponding CAS no.a ‘+’, Positive; (+), weakly positive;�, negative; ?, inconclusive; NT, not tested; without e.m.s., without exogenous metabolic system; with e.m.s., with exogenous metabolic

system.b LED, Lowest effective dose; HID, highest ineffective dose; NR, not reported; po, oral; ip, intraperitoneal; sc, subcutaneous.c The number in parentheses indicates the ratio [high animal dose (mg/m2)/maximum recommended human dose (mg/m2)].


1.4% only in mice, and the 1.4% only in hamsters and in dogs andmonkeys, respectively.

Table 3 provides for each assay type the number of pharma-ceuiticals with positive, negative and discordant results. It is worthnoting the low number of consistently positive drugs in all thethree types of DNA lesions assays, whereas 20 drugs tested positivein all the carcinogenesis assays performed, and 64 werecarcinogenic in at least one sex of mice or rats or other speciesbut gave discordant results.

Concerning the correlation among the results of the three typesof DNA lesions assays, results were discordant for the 26.7% ofdrugs between DNA strand breaks and UDS, and. constantlyconcordant between DNA adducts and DNA strand breaks andbetween DNA adducts and DNA repair synthesis but in these twocases the data available were only 2 and 4, respectively.

The results of an analysis of the correlation between the resultsof the three types of DNA lesions assays and the results ofcarcinogenicity assays in mice and rats are reported in Table 4. Thepharmaceuticals included in this analysis are those that in the DNA

Table 2Overview of DNA-damaging and carcinogenic activity of 146 marketed pharma-

ceuticals.

Drugs tested only for the formation of DNA adducts 4 (2.7%)

Drugs tested only for the formation of DNA strand breaks 31 (21.2%)

Drugs tested only for the induction of UDS 81 (55.5%)

Drugs tested for both DNA strand breaks and DNA adducts 1 (0.7%)

Drugs tested for both UDS and DNA adducts 3 (2.1%)

Drugs tested for both DNA strand breaks and UDS 21 (14.4%)

Drugs tested for DNA adducts, DNA strand breaks and UDS 5 (3.4%)

Drugs tested for carcinogenicity only in mice 2 (1.4%)

Drugs tested for carcinogenicity only in rats 13 (8.9%)

Drugs tested for carcinogenicity in both mice and rats 121 (82.9%)

Drugs tested for carcinogenicity in mice, rats and

other species

8 (5.4%)

Drugs tested for carcinogenicity only in other species 2 (1.4%)

Numbers in parentheses indicate the percentage of the 146 pharmaceuticals

considered.

lesions assays considered gave only positive result(s) or onlynegative or equivocal result(s), and in carcinogenicity assays testedpositive in at least one sex of mice or rats or gave only negative orequivocal result(s). It is evident that the percentages of discordantresults are lower for DNA strand breaks and UDS, ranging from the35.9% to the 43.4% of pharmaceuticals, than for those of DNAadducts that range from 55.6% to 66.7%, but this latter case shouldbe considered not much indicative due to the few available data.However, these percentages should be considered with cautionsince often long-term carcinogenesis assays were performed atdaily doses to various extent lower than the daily doserecommended by present guidelines for carcinogenicity testingof pharmaceuticals [119,120], that should be at least 25-fold higheron a mg/m2 basis than the maximum recommended human dailydose. If the daily doses used in long-term carcinogenesis assays areexamined, it becomes evident that they were lower than therecommended dose for 54 and 41 drugs that tested negative forDNA lesions in mice and rats, respectively.

Finally, Table 5 indicates the number of pharmaceuticals thataccording to the results obtained in the three DNA lesions assaysmay be classified as non-genotoxic non-carcinogens, genotoxicnon-carcinogens, non-genotoxic carcinogens, and genotoxic

Table 3Summary per assay type of pharmaceuticals with positive, negative and discordant

results.

Assay type Positive Negative Discordant

DNA adducts 2 (15.4%) 8 (61.5%) 3 (23.1%)

DNA strand breaks 5 (8.6%) 41 (70.7%) 12 (20.7%)

DNA repair synthesis (UDS) 7 (6.4%) 96 (87.2%) 7 (6.4%)

Carcinogenesis 20 (13.7%) 61 (41.8%) 65 (44.5%)

Pharmaceuticals considered as positive are those that gave only positive result(s).

Pharmaceuticals considered as negative are those that gave only negative or

equivocal results. Discordant indicates the number of pharmaceuticals that gave

both positive and negative or equivocal results in DNA lesions assays, and in

carcinogenesis assays performed in the same species or in different species.

Table 4Correlation between the results of DNA lesions assays and the results of carcinogenicity assays.

Couples of assays considered No. of pharmaceuticals with

Concordant results Discordant results

DNA adducts—carcinogenicity in mice 4 (44.4%) 5 (55.6%)

DNA adducts—carcinogenicity in rats 4 (40.0%) 6 (60.0%)

DNA adducts—carcinogenicity on both species 3 (33.3%) 6 (66.7%)

DNA strand breaks—carcinogenicity in mice 25 (62.5%) 15 (37.5%)

DNA strand breaks—carcinogenicity in rats 26 (59.1%) 18 (40.9%)

DNA strand breaks—carcinogenicity in both species 19 (63.3%) 11 (36.7%)

UDS—carcinogenicity in mice 60 (61.2%) 38 (38.8%)

UDS—carcinogenicity in rats 60 (56.6%) 46 (43.4%)

UDS—carcinogenicity in both species 41 (64.1%) 23 (35.9%)

The pharmaceuticals included in these comparisons are those that in DNA lesions assays considered gave only positive result(s) or only negative-equivocal result(s), and in

carcinogenicity assays tested positive in at least one sex of mice or rats or gave negative-equivocal result(s) in the species considered.

Table 5Number of non-carcinogens and of carcinogens that tested negative and positive in the different types of DNA lesions assays.

Assay type No. of non-genotoxic

non-carcinogens

No. of genotoxic

non-carcinogens

No. of non-genotoxic

carcinogens

No. of genotoxic

carcinogens

DNA adducts 1 (7.7%) 0 7 (53.8%) 5 (38.5%)

DNA strand breaks 25 (43.1%) 5 (8.6%) 16 (27.6%) 12 (20.7%)

UDS 39 (35.5%) 1 (0.9%) 57 (51.8%) 13 (11.8%)

The data indicate the number of non-carcinogens and of carcinogens examined in each DNA lesions assay that tested negative or equivocal (non-genotoxic) and positive

(genotoxic) in the same assay. In this analysis were considered: (i) as non-carcinogens the pharmaceuticals that did not increase tumour incidence in mice and/or rats and/or

other species and as carcinogens those pharmaceuticals that increased tumour incidence in at least one sex of mice or rats or other species; (ii) as non-genotoxic the

pharmaceuticals that gave a single negative-equivocal result or concordant negative-equivocal results, and as genotoxic the pharmaceuticals that gave at least one positive

result in the indicated assay.


carcinogens. It is evident that the percentage of non-genotoxicnon-carcinogens and of non-genotoxic carcinogens is dependenton the DNA lesions assay considered. For instance, are classified asnon-genotoxic carcinogens the 27.6% of pharmaceuticals thattested negative in the DNA strand breaks assay, and the 51.8% ofthose that tested negative in the UDS assay. Also in this case thefew data available for DNA adducts are not much indicative.

If we presumed that a pharmaceutical might be consideredgenotoxic if tested positive in at least one DNA lesions assay and tobe a carcinogen if tested positive in at least one carcinogenicityassay, 20 drugs (13.7% of the 146 drugs examined) might beclassified as genotoxic carcinogen. Fifty-four drugs (37.0%) thattested negative in DNA lesions assay(s) and in carcinogenicityassay(s) might be classified as non-genotoxic non-carcinogen.Sixty-six drugs (45.2%) that tested negative in DNA lesions assay(s)but were carcinogenic in at least one sex of mice or rats or otherspecies might be classified as non-genotoxic carcinogens. Six drugs(4.1%) that tested positive in at least one DNA lesions assay butnegative in carcinogenicity assay(s) might be classified asgenotoxic non-carcinogens.

3. Discussion

The guidelines for genotoxicity studies on pharmaceuticals [1–3] suggest that in addition to the standard 3-test battery it may beuseful to perform the tests on the formation of DNA adducts and onthe induction of DNA strand breaks and DNA repair synthesis.Moreover, recommendations for the performance of UDS tests invitro and in vivo have been given by the Workshop onStandardization of Genotoxicity Test Procedures held in February1993 [106]. Taking into account these suggestions, we haveexamined 146 marketed pharmaceuticals in order to establish towhat extent the results provided by DNA lesions assays arepredictive of carcinogenic activity in rodents. A positive correlationwith the presence or absence of carcinogenic activity in both miceand rats was found to be present for the 63.3% of pharmaceuticalsconsidered in the case of the DNA strand breaks, for the 64.1% in

the case of DNA repair synthesis, and for the 33.3% in the case ofDNA adducts, but the last finding is of limited significance due tothe few data available. Taking as a whole the three types of DNAlesions assays, an even higher degree of positive correlation withcarcinogenicity in mice and rats was found to be present in aprevious study on benzodiazepines [121], and in another previousstudy on analgesics, anti-inflammatory drugs and antipyretics[122], the percentage of drugs with concordant results rangingfrom 66.7% to 75.0% and from 75.0% to 100%, respectively. Sasakiet al. [123] have found that for rodent genotoxic carcinogens theresults of the DNA damage comet assay on multiple mouse organsare positively correlated with carcinogenic activity for the 94% ofthe compounds examined. According to Probst et al. [113], a surveyon 218 compounds reveals that recognized carcinogens are readilydetected by the HPC-DNA repair test whereas structurally relatednon-carcinogens produce negative results.

Concerning the comparison of the capability of DNA lesionsassays of predicting the occurrence or absence of carcinogenicitywith that of other types of assays, 136 of 472 drugs considered in aprevious review [124] were tested for genotoxicity in the standard3-test battery, for DNA lesions and for carcinogenicity. The resultsobtained are shown in Table 6. If only the drugs which gaveconcordant results in the types of assays considered are examined,for 51 drugs a correct prediction of the results of carcinogenesisassays was given by the results of DNA lesions assays, while for 25drugs the prediction was wrong; with respect to the standard 3-test battery, a correct prediction was given for 41 drugs, while theprediction was wrong for 16 drugs. Two carcinogenic drugs whichtested negative in the assays of the standard 3-test battery gavepositive results in DNA lesions assays. If also the drugs which gavediscordant results in one or both types of the two assaysconsidered are examined, since it is reasonable to assume thatalso a single positive result might be indicative of a possiblegenotoxic, DNA-damaging or carcinogenic activity, for 14 drugs acorrect prediction of the results of carcinogenesis assays was givenby the results of DNA lesions assays, while for 46 drugs theprediction was wrong; with respect to the standard 3-test battery,

Table 6Results obtained with 136 drugs examined for genotoxicity in the standard 3-test

battery, for DNA lesions, and for carcinogenicity.

No. of drugs Standard 3-test battery DNA lesions Carcinogenicity

41 � � �8 +/� +/� +/�8 +/� � �5 +/� +/� �3 +/� +/� +

15 +/� � +/�26 � � +/�14 � � +

1 � +/� +/�10 +/� � +

2 � + +

2 +/� + +/�1 +/� + �

Note: (�) negative result(s); (+) positive result(s); (+/�) discordant results.


a correct prediction was given for 38 drugs, while the predictionwas wrong for 41 drugs. One carcinogenic drug that testednegative in the standard 3-test battery tested positive for DNAlesions, but 10 carcinogenic drugs which tested negative for DNAlesions tested positive in the standard 3-test battery. If carcino-genic activities in mice and rats are considered separately, thepercentages of concordant results are the following ones: for DNAlesions, 62.6% and 56.6%; for bacterial mutagenicity, 68.6% and63.0%; for in vitro cytogenetics, 61.6% and 56.0%; for the mouselymphoma assay, 60.9% and 57.4%; for gene mutation at the hprt

locus, 59.8% and 58.9%, and for in vivo cytogenetics, 68.3 and 61.9%.Considered as a whole, the above mentioned data suggest that withthe DNA lesions assays the occurrence of a correct prediction of theresults of carcinogenesis assays is similar to that of the standard 3-test battery. At the same time they indicate that in a few cases theresults of DNA lesions assays can correctly predict the carcinogenicactivity of a drug that tested negative in the standard 3-testbattery.

The potential usefulness of DNA lesions assays is furthersuggested by the following considerations. In several cases resultsprovided by the standard 3-test battery are discordant, thediscordance occurring either among the results of a given typeof assay or among the different types of assays. Of the 472 drugsconsidered in a previous review [124], 42 gave discordant results inthe same type of assay, and 102 in the different types of assays.Moreover, 8 non-carcinogenic drugs negative in DNA lesionsassays tested positive in one or more than one assay of thestandard 3-test battery, and 9 carcinogenic drugs that testedpositive in DNA lesions assays gave negative responses in one ormore than one assay of the standard 3-test battery.

According to IARC [125], an agent is classified as probablycarcinogenic to humans (Group 2A) when there is limited evidenceof carcinogenicity in humans and sufficient evidence of carcino-genicity in experimental animals. Exceptionally, an agent may beclassified in this category solely on the basis of limited evidence ofcarcinogenicity in humans or sufficient evidence of carcinogenicityin experimental animals strengthened by supporting evidencefrom relevant genotoxicity data. An agent is classified as possiblycarcinogenic to humans (Group 2B) when there is limited evidencein humans in the absence of sufficient evidence in experimentalanimals. It may be also classified in this category when there isinadequate evidence of carcinogenicity in humans or when humandata are non-existent, but there is sufficient evidence ofcarcinogenicity in experimental animals. In some instances anagent for which there is inadequate evidence or no data in humansbut limited evidence of carcinogenicity in experimental animalstogether with supporting evidence from other relevant data maybe placed in Group 2B. On the basis of these indications, danthron,

that gave consistent positive results in the DNA lesions assays, wasclassified by IARC as possibly carcinogenic to humans [34]. Now itmight be classified in the same group also hydralazine previouslyjudged by IARC not classifiable as to its carcinogenicity to humans[125].

In conclusion, the information reported by this survey can beconsidered as indicating that for the assessment of the potentialgenotoxic-carcinogenic risks to humans the DNA lesions assaysmight be a useful addition to the standard 3-test battery.

Conflict of interest

None.

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