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2 (1) (2) (1) (2) (1) Seg (2) Seg (3) Seg (4) Seg (1) (2) (3) ( ) (1) (PCA) (2) Draize (1) Ames in vitro in vivo (1) (2)...138

3 2-AA 9-AC ALP EMS DNCB ENNG OTC LD 50 NF 20-MC CPA MNPCE MCH MCHC PCE S.D. S.E N- -N - -N- ( ) 50% Micronucleated Polychromatic Erythrocytes Mean Corpuscular Hemoglobin Mean Corpuscular Hemoglobin Concentration Polychromatic Erythrocytes ( ) ( ) (+)-(S)-4-[1-(2,3-dimethylphenyl)ethyl]-1 H-imidazole monohydrochloride JAN dexmedetomidine hydrochloride (DEX) INN dexmedetomidine (R)-4-[1-(2,3-dimethylphenyl)ethyl]-1H-i midazole

4 -1 (g/kg/) (g/kg/) : 5 mg/kg mg/kg : 10 mg/kg : 5 mg/kg mg/kg : 5 mg/kg : 2 mg/kg mg/kg : >2 mg/kg GLP ( ) -1 ( ) -2-3 ( ) -4-5 ( ) : < : <10 ( ) : < : <10 ( ) 10 6 Seg (2 ) ( ) ( ) Seg < ( ) Seg ( ) 8 16 ( ) g/kg/infusion ( ) ( ) ( ) g/kg/infusion 1.0 mg/animal -13 (6 ) ( ) 50g/animal -14 (10 ) ( ) 106

5 51500 g/plate ( ) Ames Test g/plate ( ) g/ml ( ) GLP ( ) g/ml S9 mix g/ml -18 ( ) in vitro g/ml -19 S9 mix g/ml ( ) g/kg ( ) NMRI 5.0 mg/kg ( ) -1 in vivo CD-1-21 mg/kg ( ) g/mL -22 g/ml ( ) g/mL in vitro -23 g/ml ( ) g/kg/ : 400 : 400 g/kg/ ( ) mg/kg 2 mg/kg 1 mg/kg g/kg/ 10g/kg/ g/kg/ 250g/kg/ 10g/kg/ 107

6 18g/kg/ 54g/kg/ 6g/kg/ 200g/kg/ 20g/kg/ 96g/kg/ 96g/kg/ 8g/kg/ 2g/kg/ ( ) Ames 5 mg/kg NMRI GLP CD-1 GLP 5 mg/kg L 28 PR 400 g/kg/ 108

7 0.120 mg/kg mg/kg 10 5 mg/kg NMRI 30 g mg/kg 5 5 mg/kg 5 mg/kg : mg/kg 10 mg/kg : 1 20 mg/kg : 1 SD mg/kg 5 5 mg/kg 5 5 mg/kg 5 mg/kg : 4 10 mg/kg : 5 20 mg/kg : 5 5 mg/kg : 4 10 mg/kg : 4 20 mg/kg : 5 109

8 mg/kg 14 ( -3) 2.0 mg/kg 1 1 ( ) mg/kg 2.0 mg/kg mg/kg 2.0 mg/kg mg/kg 2.0 mg/kg 1015kg mg/kg 0.1mL/kg mg/kg 1.0mg/kg 2.0mg/kg ( ) () ( ) ( ) () ( ) ( ) ( ) 2 mg/kg 110

9 1 10 SD g/kg/ g/kg/ 160g/kg/ 40g/kg/ g/kg/ 160g/kg/ 160g/kg/ MCH MCHC 10 g/kg/ 40g/kg/ ALP 160g/kg/ PT PTT 40g/kg/ 1) 40g/kg/ 160g/kg/ ALP ALP ALP 2 2) 160g/kg/ 40g/kg/ 3)-5) 111

10 10g/kg/ 10 /kg/ SD g 150 g 1 ml/kg (g/kg/) RBC Hb Ht Ht MCH MCHC Na + ALP ALP K +, Mg +, Na + ph ph ph ( ) ( ) ( ( ) ) <10g/kg/ Bonferonni t 1) Kaheinen, P.: Research Report , Farmos Group Ltd., Research Center 2) Hirsimaki, P. et al: Research Report , Farmos Group Ltd., Research Center 3) Kamibayashi, T. et al:anesthesiology 93: (2000) 4) Bloor, B.C. et al: Anesthesiology 77: (1992) 5) Schmeling W.T. et al: Anesthesiology 75: (1991) 112

11 g/ g/kg/ 250g/kg/ 50g/kg/ 50g/kg/ 250g/kg/ 10 ( ) 250g/kg/ g/kg/ OTC( ) 250g/kg/ GPT OTC GPT 1) 2),3) 50g/kg/ PAS (Intracytoplasmic eosinophilic inclusion bodies) 250g/kg/ 1 PAS 4)-7) 113

12 10g/kg/ 10g/kg/ kg kg 0.25 ml/kg (g/kg/) OTC OTC GPT ph ph * 10g/kg/ * OTC(Ornithine Transcarbamylase) 1) Amacher, D.E. et al: Food Chem Toxcol 36: (1998) 2) Sanhouri, A.A. et al: Br Vet. J. 146: (1990) 3) Yannai, S. et al: Arch Toxicol 54: (1983) 4) Ghadially, F.N.: Fourth Ed., Boston: Butterworth-Heinemann, pp , (1997) 5) Wills, E.J.: Ultrastructural Pathology 16: (1992) 6) Biava, C. et al: Am J Pathol 46: (1965) 7) Murti, G.S. et al: Am J Vet Res 26:63-67 (1965) 114

13 24 SD g/kg/ g/kg/ 54 / 18g/kg/ 18g/kg/ 6 g/kg/ 18 54g/kg/ 18g/kg/ 20 18g/kg/ 54g/kg/ 2 1), 2) 54g/kg/ g/kg/ 2 3) F1 18g/kg/ (F0) 18g/kg/ 6g/kg/ 54g/kg/ (F1) 18g/kg/ 6g/kg/ 1)Magness, R.R. et al : Am. J. Obstet. Gynecol , )Eisenach, J.C. et al : Am. J. Obstet. Gynecol. 160(2) , )Akatuka, K. et al : J. Toxicol. Sci.(Suppl 2)

14 ) SD 6~7 210 g 10~ g 1 ml/kg g/kg/ (%) 2) 100 (24/24) 91.7 (22/24) 100 (24/24) 100 (24/24) (%) 2) 100 (24/24) 81.8 (18/22) 95.8 (23/24) 95.8 (23/24) ,3,4) ,3,4) (%) ,2,4) (%) ,2,4) ,2,4) ,3,4) ( ) (g) 3,4) ** *** (100 ) (79 ) (77 ) (97 ) (%) 100 (12/12) 100 (8/8) 100 (13/13) 91.7 (11/12) () 4) , , , ** *** (%) (%) * *** (g) 4) *** *** (%) 90 (9/10) 100 (9/9) 100 (13/13) 92.3 (12/13) 1,3, (%) 1,3,4) F 2 4 (%) 2) (g) 4) g/kg/ 54g/kg/ 6g/kg/ (1) 1 2) Fisher 3)Bonferonni t 4) S.D. * p<0.05 ** p<0.01 *** p<

15 25 SD g/kg/ g/ / 200 g/kg/ 20g/kg/ 200g/kg/ g/kg/ 200g/kg/ g/kg/ Seg 2 20g/kg/ 2g/kg/ 200g/kg/ 20g/kg/ 200g/kg/ 20g/kg/ 117

16 SD 3 ( ) g 1 ml/kg (g/kg/) ) 1,3,4) ,3,4) ,3,4) (% *** 2,3,4) ** 2,3,4) ) )1) *** (g) 1,4) *** (g) 1,4) *** ( ) 1) g/kg/ 20g/kg/ 20g/kg/ 1) Bonferroni t 2) 2 Fisher 3) * p< 0.05,** p< 0.01, *** p< ) S.D. 118

17 12 Himalayan g/kg/ g/kg/ 60 24g/kg/ g/kg/ 24g/kg/ g/kg/ g/kg/ 6g/kg/ 6g/kg/ 96g/kg/ 96g/kg/ Himalayan kg ml/kg g/kg (g/kg/) ) 1,3,4) ,3,4) () 2) % ,3,4) ) hr hr ) (g) 1,4) (g) 1,4) ( ) 2) (%) <6g/kg/ 96g/kg/ 96g/kg/ 1)t 2)Fisher 3)1 4) S.D. 119

18 1 22 SD g/kg/ Seg Seg 2g/kg/ 32g/kg/ 8g/kg/ 8g/kg/ 32g/kg/ 16g/kg/ 4 32 g/kg/ 8g/kg/ 7 (16 ) 32g/kg/ 2 32g/kg/ 20 32g/kg/ 8g/kg/ 2g/kg/ 120

19 SD ~240 g 1 ml/kg g/kg/ ) (%) 100 (22/22) 95.5 (21/22) 100 (22/22) 100 (21/21) 4) 1,3,7) (%) ,3,7) (%) (g) ( ) ) (%) 100 (21/21) 100 (21/21) 100 (21/21) 100 (21/21) (%) 95.2 (20/21) 100 (21/21) 100 (21/21) 90.5 (19/21) 1,3,7) ,3,7) ,3,7) ,3,7) ,3,7) F 2 (g) 7) (g) 7) g/kg/ 2g/kg/ 2g/kg/ 1) t 2) 2 Fisher 3) 1 4) 1 5) 6) 7) S.D. * p<0.05 ** p<0.01 *** p<

20 g/kg 416g/kg g/kg 0.25g/kg 4g/kg 8g/kg 4g/kg 8 16g/kg SD 116g/kg g/kg SD 8 16g/kg/ 4 mg/kg/ (168 ) 2 ml/kg/ mg/kg ) g/kg 122

21 p0.05, (Mann-Whitney U ) g/kg ( -10) hr 5hr p0.05 (Dunnett ) 123

22 No. 1hr 5hr S702 S705 S777 2mL/kg/infusion S786 S789 S793 L776 L779 L780 L790 8g/kg/infusion L791 L796 H701 H769 H782 16g/kg/infusion H783 H784 H785 P700 P767 P770 4mg/kg/infusion P792 P794 P797 : : ++ :

23 6 ( kg) 3 mg/kg mg/kg g/kg (16 24 mg/kg) 2 Seevers (A) (B) (1) (%)100 AB A 3 1 No. (%) 0.5 ml/kg 0.25 ml/kg g/kg g/kg mg/kg 24 mg/kg ) ) Seevers, M.H. : J. Pharmacol. Exp. Ther. 56, ,

24 4 ( kg) 10 / g/kg/ 30 / g/kg g/kg 30 / g/kg/ g/kg g/kg * (1 )** (2 )** (g/kg) 125 or or 1.0 No or 3 2 or * 3 ** 1 No g/kg (2 ) 0.25g/kg No

25 PCA Draize Hartley ( g 6 /) 2 ml(t 0 : ) 1.0 mg/2 ml/animal (T 1 : ) 1.0 mg/2 ml/animal (T 2 )1 3 2 (6 ) ml (T 0 T 1 T 2 T 3 T 4 T 5 ) 45 T 3 T mg /ml/animal T mg/ml/animal 0.5% (T 4 ) (T 3 ) (T 5 ) T 0 T 1 1) 2) PCA / s.c. T 3 (0.1 mg/animal) 5) i.v. 06 (1.0 mg/animal) 3) s.c. T 4 (1.0 mg/animal) 6) i.v. 66 T 2 (1.0 mg/animal) 4) s.c. T 5 (0.1 mg/animal) 5) i.v. 06 1) 3 2 (6 ) 2) PCA 1/16 1/64 1/ ) (0.5 mg/ml)2ml # 4) (0.5 mg/ml)2 ml # 5) (0.1 mg/ml)1 ml 6) (1.0 mg/ml)1 ml # 127

26 Hartley ( g 6~10 /) (T 0 : ) 0.05(w/v)DNCB(T 1 : ) 0.05 # (T 2 ) 1 3 (10 ) (1 0.1 ml/animal ml/animal) 2 T DNCB T 0 T ml DNCB (T 1 ) (T 0 T 2 ) ( -14) / No. T 0 T 1 T 2 / / DNCB /DNCB / (24 48 ) (24 48 ) 4 0/6 3/6 0/10 # 128

27 Ames in vitro in vivo TA1535 TA1537 T1538 TA98 TA g/ 5000g/ g/ g/ g/plate 1) 5 5g/plate 6 ( -15) 1500g/ g/ WP2uvrA g/ g/ 7 ( -16) 11000g/plate 1) OECD Guideline for testing of chemicals No. 471: Bacterial reverse mutation test 129

28 / TA1535 TA1537 TA1538 TA98 TA100 (g/ ) S-9mix (-) S-9mix (+) ENNG IL IL IL IL IL IL AC 80.0 X X NF AA IL IL IL IL S.D. N=3 ENNG N- -N - -N- 9-AC 9- NF 2-2AA 2- IL X 130

29 WP2uvrA / (g/ ) S-9mix (-) S-9mix (+) ENNG AA ENNG N- -N - -N- S.D. N=3 2AA 2-131

30 L5178Y TK+/-TK-/- L5178Y S-9mix TK % 200g/mL 10150g/mL S-9mix 2 S-9mix S-9mix 40g/mL 20200g /ml S-9mix 40g/mL 22237/10 6 cells 1) ( -17) (g/ml) () 1 2 () S-9mix () S-9mix () DMSO EMS *** *** DMSO * MC *** *** EMS 20-MC cell *p0.05, ***p ) Mitchell I.deG. et al: Mutagenesis 5: (1990) 132

31 18 32 S-9 mix % 3 S-9mix (1 300g/mL 2 350g/mL 32 ) % ( -18) S-9mix S-9mix S-9mix(-)EMS( ) S-9mix(+)CPA( ) (g/ml) S-9mix g *** 18 () (%) g *** 1 + (g/ml) S-9mix g * 20.0*** () (%) g * 20.5*** + (g/ml) S-9mix g *** 18 () (%) g *** + (g/ml) S-9mix g *** () (%) g *** 2 + (g/ml) S-9mix g *** 32 () (%) g *** + (g/ml) S-9mix g * 37.5*** () (%) g * 38.0*** (g) (+g) ***p0.001, *p

32 S-9mix -18 S-9mix 1 168g/ml 488g/ml g/ml g/ml 3 19 C B1-19 S-9mix 3 1 S-9mix S-9mix (-) C S-9mix (+) B1 S-9mix () S-9mix () 19 S-9mix () 2 3 S-9mix () 100 * p0.05 (g/ml) (%) * + (g/ml) (%) * + (g/ml) (%) * + (g/ml) * (%) + 134

33 NMRI g/kg PCE/NCE 2000 PCE PCE MNPCE g/kg -20 PCE/NCE MNPCE 250g /kg NMRI 5 mg/kg GLP -1 GLP NMRI g ( 24 ) 40( /kg) PCE/NCE MNPCE (%) PCE/NCE ( /kg) 250( /kg) 40(mg/kg) ** ** ** MNPCE (%) MNPCE(Micronucleated Polychromatic Erytrocytes) MNPCE(%)=MNPCE /PCE 100 **p0.01 1) O brien, H. et al: Br J Haematol 51: (1982) 2) Asanami, S. et al.: Mutation Res. 393: (1997)

34 CD mg/kg % 1ml PCE/NEC MNPCE (%) 50 mg/kg PCE/NECMNPCE (%) PCE/NEC MNPCE (%) CD (m /kg) 2.5(m /kg) 5.0(m /kg) 50(mg/kg) PCE/NCE * MNPCE (%) ** 24 PCE/NCE MNPCE (%) PCE/NCE * MNPCE (%) ** 48 PCE/NCE MNPCE (%) MNPCE(Micronucleated Polychromatic Erytrocytes) MNPCE(%)=MNPCE /PCE 100 *p0.05, **p

35 5 SD g/mL( 0.1mL) 5 20 mg/ml g/mL g/mL 1000g/mL 1 10 SD 400g 1 5 ( g/ ml 0.1mL 5 20mg/mL 0.1mL 0.1mL (g/ml) 20mg/mL

36 g/mL in vitro ( ) ( ) 545 nm g/mL 3000g/mL 100% ( ng/mL*) *: DEX W97-249W97-245W97-246W99-302W J-DEX mL 100L nm (g/ml) (%) g/kg/ 10000g/kg g/kg 2000g/kg 1 PR g/kg g/kg g/kg 10000g/kg 400g/kg/ 138

37 kg g (1 :3 :3 ) 400, 2000, 10000g/kg/ g/kg/ 3 2 (g/kg/) ) 2) PR 1) 2) 2) 400g/kg/ 1) 2) 139

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