### BII-0908

Table 14·4 Twenty-eight-day repeated-dose oral toxicity study in rats Summaryofhistopathological examinations Male Female
Findings Grade Vehicle control Vehicle control (R )

ecove!l 25 ISO 1,000 1,000 (Recovery) Vehicle control Vehicle control (Recovery) 25 150 1,000 1,000 (gfkgfd ) (Recovery) m . ay
ta ta ta Ia fa fa fa ta Ia III III fa 51' S 5 j 5 S 5 5 S S 5 S 

### Axillar lymph node

No abnormalities detected 515bl 515 515 515 Mesenteric lymph node

No abnonmalities detected 515 515 515 515 Spleen

No abnormalities detected 5/5 5/5 5/5 5/5 Thymus

No abnormalities detected 515 5f5 515 515 <0 I No abnormalities detected 515 SIS 5/5 515

-----------­ Thyroid

No abnormalities detected 5f5 5f5 515 SIS PlInIlhyroid

No abnonnalities detected 515 5/5 5/5 515 Adrenal

No abnonnalities detected 4/5 5/5 5/5 515

................................................................... _ ....................................... _ ... __ .... __ ...... ___ ...................... #0
...................................................... " ...... - ... _ ..... -- .......................... _ .... F()Cal hypertrophy ofcortical 

### + If5 015 0/5 0/5 

### cells 

### Eyeball

No abnormalities detected 

### Sf' 515 !filS SIS Skin

Scab [onmation 

## + 1/1 ta, tenmioal autopsy. a) Numberofanimals autopsied. b) Numberofanimals affected INumber ofanimals examined. A -,Not examined. W CJ1 +, slight.B11-0908
Addendum 1-1 Twenty-eight-dayrepeated-doseoral toxicity studyinrats Clinical signsofindividual animals (males) Exp. group Animal Administration Period (day) RecoveryPeriod (day) (mglkgfday) No. 

### Sings 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 1920 21 22 23 24 25 26 27 28 I 2 3 4 5 6 7 8 9 10 II 12 13 14

2 Vehicle 

### 3 control 

### 4 

# , 6 Vehicle 7 control 

### 8 (Recovery) 9
I 00 

### o I 10

-: No abnormalitiesdetected.



### w ..j:lo. c::J)BII-0908
Addcndum 1-2 Twenty-eight-dayrepeated-dose oraltoxicity study in rats Clinical 

### signs ofindividual 

### animals (males) Exp. group Animal Administration Period (day) (mglkglqay) No. 

### Sings 2 34 S 6 1 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 21 28 1l 12**---------~-----~-------.-"'-.--,,-.----------** 2S 13 

### 14 

### IS
-:No abnormalitiesdetected.

I 00 I-' I

W -1:>0 --.J

## ----------------------------------------------- B11-0908
Addendum 1·3 Twenty-eight-day repeated-dose oral toxicity study in rats Clinical signs ofindividual animals (males) 

### Exp_ group Animal Administration Period (day) (mglkglday) No. Sings I 2 3 4 

### 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 16 Salivation 

### 15 17 20 21 Decreased spontaneous 16 

### 20 21 22 locomotion 17 

### 150 Salivation 

### 16 18 21 

### 22 24 25 26 28 18 Salivation 10 13 14 15 16 17 

### 19 20 

### 21 22 23 24 25 27 19 Salivation 3 4 

### 8 

### 10 13 14 

### 15 16 

### 17 18 19 22 

### 23 24 26 27 28 1 00 20 N 1 Salivation 13 15 16 17 26 27 28 Decreased spontaneous locomotion 

### 24

.:No abnormalities detected.

(..)

### ~

### B11-0908
Addendum 1-4 Twenty-eight-day repeated-dose oral toxicity study in rats Clinical signs ofindividual animals (males) Exp. group Animal Administration Period (day) 

### (mglkgfday) No. 

### Sings 2 3 

### 4 S 

### 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 

### 21 Salivation 

### 2 3 

### 4 5 6 7 

### 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 Decreasedspontaneous locomotion 6 

### 7 8 9 12 13 14 15 16 17 21 22 24 25 26 27 28
22 Salivation 

### 3 4 5 6 

### 7 8 

### 9 10 II 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 Decreasedspontaneous locomotion 

### 7 9 13 14 16 17 18 19 21 22 23 24 27 28

I 00 

### c.u I Moistening ofmandible Moistening ofmandible and neck 

### 1,000 Loss ofhair(mandible) Muchfood on tray Lacrimation 

### 23 Salivation 

### 2 3 4 :5 

### 6 7 8 

### 9 10 II 13 

### 15 17 18 21

19 20 19 20 21

19 20

12 13 14 15 16 17 18 19 20 21 22 23 26 27 28

25
22 23 24 25 26 27 28 22 25

22 23 24 25 26 27 28 Decreased spontaneous locomotion 

### 7 9 10 12 15 16 17 19 20 21 22 23 Incomplete eyelid opening 7
Moisteningofmandible 

### 19

C.:I ..lIoo 

### co Muchfood on tray
-: No abnormalities detected. 

### 21

### BIl-0908
Addendum 1-5 Twenty-eigbt-day repeated-dose oral toxicity study in rats Clinical signs ofindividual animals (males) Exp. group Animal Administration Period (day) (mglkg/day) No. Sings 

### 2 3 4 5 6 7 8 9 IQ II 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28

I 00 If::o I 

### 24

1,000

25 Salivation Decreased spontaneous locomotion Moistening

### ofmandible
lollS

### ofhair(mandible) Much food on tray

Salivation Decreasedspontaneous locomotion Much food on tray
-: No abnormalities detected. 

### 3



### 3 4



### 4 5



### 5 6



### 6 7



### 7 8



### 8 9
9



### 9 10 Jl

10 11 12 13
13



### 12 

### 13
13 14 15
14

15



### 14 15
14 15 16 17 18 19 20 21 20 21

18 21 

### 19 20 21 21



### 16 17 

### 18 19 20 21
16 17 21 22 23 24 25 26 27 28 24 25

22
22 23 24 25 26 22 23 24 25 26 27 28



### 22 23 24 

### 25 26 27 28
22 23 24 26
23 25 27 28

(,.) t.n 

### oBlI·0908 Addendum 1-6 Twenty-eight-day repeated-dose oral toxicity study in rats Clinical signs ofindividual animals (males) Exp. group Animal Administration Period (day) Recovery Period (day) (llIJIk&Idlly) No. 

### Sings 2 3 4 5 6 7 8 9 10 II 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 2 3 4 5 6 7 8 9 10 II 12 13 14

I 00 t;;"J I 1,000 (Recovery) 

### 26

27 Salivation Decreased spontaneous locomotion Loss afhair(mandible) Much food on tray Moistening ofmandible

Salivation Decreased spontaneous locomotion 

### Loss ofhair(mandible) 

### Loss ofbair(mandible and neck) 

### 3

3 

### 4

4 

### 5

5 

### 6
6

6 

### 7
7

7 

### 8

8 

### 9

9 

### 10 II 11

10 II
11 12
12

12
12
12 

### 13 14 15 16 17 18 

### 13 14 

### 15 16 17 18

13 14 15 16 17 18
13 14
13 14 15 16 17 18 192021 22 23 

### 19 21
19 20 21 22 23 22
23

19 20 21 22 23
19 20 21 22 23
19 20
20 21 22 23 24 25 2627 28
24 20 27 28

24 25 26 27 28
24 26 27 28

24 25 26 27 28 

### I 2 3 4 5 6 7 8 9 10 11
Moistening ofmandible 15 17 18 26 Moistening ofmandible and neck

-:No abnormalities detected. 20 21 22 23 25 27 28

w (J'IB1I-0908 Addendum 1-7 Twenty-eight-day repealed-dose oral toxicity study in rats Clinical signs 

### ofindividual 

### animals (males) Exp. group Animal Administration Period (day) Recovery Period (day) (mg/kglday) No. Sings 2 3 4 .5 6 7 8 9 10 II 12 13 14 15 16 17 18 192021 22 23 24 25 2627 28 2 3 4 5 6 7 8 9 10 II 12 13 14 21 Salivation Decreasedspontaneous locomotion 2 3 4 5 6 7 8 9 

### to II U
12 13 14 15
14 16 17 18 19 20 21
19 20 21 22 23 24 25 26 27 28
22 24
Moistening ofmandible \3 15 17 18 21 22 23

I 00 0':> I 

### 1,000 (Recovery) 

### 29 Moistening ofmandible and neck Much food on tray Loss ofhair(mandible)

Salivation Decreased spontaneous locomotion 3 4 5 6 7 8 9 10
10 11
II 12
12 13
13 14
14 15
IS 16
16 17 18
17 19 20
21

1920 21
19 20 25
23 24 26 22 23

22 23 24 25 2627 28
22 23
Moisteningofmandible 25
30 Salivation J 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 1920 21 2223 24 25 26 27 28 Decreased spontancous locomotion 10 13 14 15 16 17 18 23 24 25 26 27 28
Much food on

### tray 20 21 22 23 25 27 3 -; No abnormalities detected.

(,..) <.n N811-0908
Addendum 1-8 Twenty-eight-dayrepeated-dose oral toxicity study in rots Clinical signs ofindividual animals (females) Exp. group Animal Administration Period (day) Recovery Period (day) (mglkglday)

. No. Sings 2 3 4 5 6 7 8 9 10 II 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 I 2 3 4 5 6 7 8 9 10 II 12 I3 14 

### 31 

### 32 Vehicle



# -- 33 control 34



## - 35 

### 36



# -- 37 Vehicle control 

### 38 (Recovery) 39

- 40 I 00 -: No abnormalities detected. -...l I



### w U1 WBll-0908
Addendum 1-9 Twenty-eight-dayrepeated-dose oral toxicity studyin rats Clinical signs ofindividual !IIlimals (females) Exp. group Animal AdministrationPeriod (day) (mglkglday) No. Sings 

### 2 3 4 

### S 6 7 8 9 10 11 12 13 14 

### IS 16 17 18 19 20 21 22 23 24 

### 2S 26 27 28 41 42 

### 25 43 44 45

-: No abnormalities detected.

I CO CO I

w c.n -1:>0BII-0908
Addendum 1-10 Twenty-eight-dayrepeated-do5e oral toxicity studyinrats Clinicalsigns ofindividual anima.ls (females) Exp. group Animal AdministrationPeriod (day) (mglkgfday) No. Sings 2 3 4 S 6 1 8 9 10 11 11 

### \3 14 IS 16 17 1& 19 20 2J 22 23 2il 25 26 27 28 46 

### Salivation 

## 24 47
150 

### 48 Salivation 3 4 9 10 II 12 13 24 25 2627 28

Salivation 

### 3 13 

### 49 

### so
-: No abnorma.lities detected.
I 00 to I

W 0'1 0'1B11-0908
Addendum I-II Twenty-eigbt-dayrepeated-dose oral toxicity study in rats Clinical signs ofindividualllIlimals(females) Exp. group Animal Administration Period (day) (mg/kg/day) No. Sings 1 2 3 4 .5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 51 Salivation 1 2 3 4 5 6 7 8 9 }O II 12 13 14 }5 16 17 18 19 20 21 22 23 24 25 26 27 28 Decreased spontaneous locomotion 10 11 14 16 17 28 Incompleteeyelid opening 10

Muc:h food ontray 24
52 Salivation 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 I 1,000 1:0 Decreased spontaneous 

### o 6 8 II· 13 14 17 21 22 23 24 25 I locomotion Staining lower abdomen 26 27 28 53 Salivation Staininglowerabdomen Decreased spontaneous locomotion 3 4 4 5 5
5 6 6

6 7 7

7 8 8
8 9

9 10

10 11 12 13

12 J3 14

14 15

15 16

16 17

17 18 18 19 20 21 20
20 21 22

22 24 25

24 25 26 27 28 26 27 28
27

Much food ontray 21 22 23 25 26 27

-: Noabnormalities detected.

(.) 0) 01

### Bll-0908
Addendum 

### 1-12 Twenty-eight-dayrepeated-dose oral toxicity study inrats Clinical signs ofindividual animals (females) Exp. group Animal Administration Period (day) (mglkglday) No. Sings 

### 2 3 4 5 

### 6 7 8 9 

### 10 II 

### 12 13 14 15 16 17 18 19 20 21 22 23 

### 24 25 26 27 28 54 Salivation 

### 2 3 

### 4 :5 6 

### 7 8 

### 9 

### 10 11 12 13 14 

### 15 16 

### 17 18 19 

### 20 21 22 23 24 25 26 27 28 Decreased spontaneous locomotion 5 

### 7 9 

### 10 II 

### 12 J3 

### 14 15 16 17 23 24 2:5 

### 26 27
Moistening ofmandible 13 

### 19

1,000 Much food ontray
Loss ofhair (mandible) 

### 20 21 20 21 22 23 

### 22 23 24 25 

### 26 27



# , (.0



## ...... I 

### 55 Salivation Decreasedspontaneous locomotion 

### 3 4 5 6
6 

### 7
7 

### 8 9 10 11 12 13 14 15 16 17 18 19 20 21
14 15 16 17 

### 22 23 24 25 26 27 28
27 28 Staininglowerabdomen 

### HI 19 20 21 26
Much food on tray 

### 22

-: No abnonnalitiesdetected.

W 01

.......BII"()908
Addendum ]-13 Twenty-eight-day repeated-dose oral toxicitystudy in rats Clinical signsofindividual animals (females) Exp. group Animal Administration Period (day) Recovery Period (day) (mglkgfday) No. Sings 2 3 4 5 6 7 8 9 10 11 12 \3 14 15 16 17 18 1920 21 22 23 24 25 26 27 28 2 3 4 5 6 7 8 9 10 II 12 13 14 56 Salivation Staining lower abdomen 

### Decreased spontaneous locomotion 3 4 4 5

5 6 7



### 7 8 9

9 10

10 11 12 13

13 14

14 15 16 17

16 17 18 19 20 21

19 22 23 24 25 26 27 28 26
24 

### Incomplete 

### eyelid opening 

### Lossofhair

### (mandible) 9 10
11 12 13 14 15 16 \7 18 1920 21 22 23

I <.0**~** I 1,000 (Recovery) Moistening ofmandible



### Muchfood on tray 13 18 19 20 21

20 23 24 25 26

22 23 24 25 26 :I
57 SaJivation Staininglower abdomen 

### Decreased spontaneous locomotion 2 3 4 5 6 7 8 9 10 II 11 12 13

12 14 15 16 17 18 19 20 21 20
21 22 23 22
22 23 24 25

24 25 26 27 28 26 27 28
26 27 28


### Moistening ofmandible 

### 27

Much food on tray 24

-: No abnormalities detected,

(.) CO 01811-0908
Addendum 1-14 Twenty-eight-dayrepeated-dose oral toxicity study in rus Clinical signsofindividualanimals (females) Exp. group Animal Administration Period (day) RecoveryPeriod (day) (mglkgfday) No. Sings 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 2 3 4 5 6 7 8 9 10 II 12 13 14 

### 58 Salivation Decreased spontaneous locomoticm 2 3 4
4 5
5 6 7 8
6 7 9
9 10
10 11
11 12
12 13
13 14
14 15 16
16 17
17 18 19 20 21
20 21 22 23 24 25 26 27 28
22 23 24 26 27 

# - - 

# - .

Incomplete eyelid opening Loss ofhair(mandible) 

### 6
11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 Moisteningofmandible 17

I**~** 

### <;=' Much food on tray 1,000 Staininglower abdomen (Recovery) 59 Salivation Decreased spontaneous locomotion Staininglower abdomen Incomplete eyelid opening 2 3 4
4
4 5

5 678 9 10 II
II

II 12 13
13 14 15 16 17 18
18 20

19 20 21
20 2J 28

26

22 23 24 25 26 27 28
24 25 28 

# . . . 

# . 

# - - -

Moisteningofmandible 15
Loss ofhair (mandible) 16 17
Muchfood on tray 21 22

w CJ1 m -; No abnonnalities detected.BII-0908
Addendum 1-1.5 Twenty-eight-dayrepeated-dose oral toxicity studyinrats Clinical signs ofindividual animals (females) Exp. group Animal Administration Period (day) Recovery Period (day) (mglkglday) No. Sings 2 3 4 S 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 2 3 4 5 6 7 8 9 10 11 12 13 14 60 1,000 (Recovery) Salivation Decreased spontaneous locomotion 2 3 4 S 6 7 8 9 10 11
10 12 13 14
14 15
1.5 16
16 17
17 18 19 20 21 22 23 24 25 26 27 28
24 26 27 Staininglowerabdomen 26
-: No abnormalities detected.

I <0**~** I**~** en 

### oAddendum2-\ Twenty-eight-day repeated-dose oral toxicity study in rats BIl'()908 Detailed clinical observations ofindividual animals (Predosing)

Exp.group Removal from cage Sex AnimalNo. (mglkglday) Ease ofremoval Vocalization 0 0 2 0 0 3 0 0 4 0 0 Vehicle 5 0 0 control 6 0 0 7 0 0 8 0 0 9 0 0 10 0 0 11 0 0 12 0 0 25 13 0 0 14 0 0 Male 15 0 0 16 0 0 17 0 0 150 18 0 0 19 0 0 20 0 0 2] 

# ° 0 22 

### 0 0 23 

# ° ° 24 0 0 1,000 25 26 0 

### 0 0 0 

### 27 0 0 28 

### 0 0 

### 29 0 0 30 

### 0 

### +1



## -95­ 361Addendum 2-2 Twenty-eight-day repeated-dose oral toxicity study in rats BII-0908 Detailed clinical observations ofindividual animals (predosing)

Exp.group Removal from cage 

### Sex Animal No. (mglkg/day) Ease ofremoval Vocalization 31 0 0 32 0 0 33 0 0 34 0 0 Vehicle 35 0 0 control 36 0 0 37 0 0 38 0 0 39 0 0 40 0 0 41 0 +1 42 0 0 25 43 0 0 44 0 0 Female 45 0 0 46 0 +1 47 0 0 150 48 0 0 49 0 0 50 0 0 51 0 0 52 0 0 53 0 0 54 0 0 1,000 55 0 0 56 0 0 57 0 0 58 0 0 59 0 0 60 0 0



## -96­ 362Addendum 2·3 Twenty·eight-day repeated-dose oral toxicity study in rats Bll-0908 Detailed clinical observationsofindividual animals (week I)

Exp.group Removal from Sex AnimalNo. (mglkglday) Ease ofrernoval Vocalization 1 0 

### 0 2 0 0 3 0 0 4 0 0 Vehicle S 

### 0 0 control 6 0 0 7 0 0 8 0 0 9 0 0 10 0 0 11 0 +1 12 0 0 25 13 0 0 14 0 0
Male 15 0 0 16 0 0 17 0 0 150 18 0 0 19 0 0 20 0 0 21 0 0 22 0 +1 23 0 0 24 0 0 1,000 25 26 0 0 0 0 27 0 0 28 0 0 29 0 0 30 0 0



## -97­ 363Addendum 2·4 Twenty-eight-day repeated-dose oral toxicity study in rats 

### 811-0908 Detailed clinical observations ofindividual animals (week t)

Exp.group Removal from cage Sex Animal No. (mglkglday) Ease ofremoval Vocalization 31 0 

### 0 32 

### 0 0 33 0 

### 0 34 0 0 Vehicle 

### 35 0 0 control 

### 36 0 

### 0 37 0 

### 0 38 0 0 

### 39 0 0 

### 40 0 

### 0 41 0 0 42 0 

### 0 25 

### 43 0 

### 0 44 0 

### 0
Female 45 0 

### 0 46 0 +1 47 0 0 

### 150 48 0 0 

### 49 0 

### 0 50 

### 0 0 51 

### 0 0 52 0 0 53 

### 0 0 54 

### 0 0 1,000 

### 55 56 0 0 0 

### 0 57 0 0 

### 58 0 0 59 0 0 60 0 0



## -98­ 364Addendum 2-5 Twenty-eight-dayrepeated-dose oral toxicity study in rats 811-0908 Detailed clinical observations ofindividual animals (week 2)

Exp.group Removalfrom cage Animal No. (mglkglday) Ease ofremoval Vocalization I 0 0 2 0 0 3 0 0 4 0 

# ° Vehicle S 0 0 control 6 0 0 7 

# ° 0 8 0 0 9 0 0 10 0 0 II 0 0 12 0 0 25 13 0 0 14 0 0 IS 0 0 Male 16 0 

# ° 17 

# ° 0 150 18 -I 0 19 0 0 20 

# ° 0 21 0 0 22 0 0 23 0 0 24 0 0 ],000 25 

### 26 0 0 0 0 27 0 0 28 0 0 

### 29 0 0 30 0 +1



## -99­ 365Addendum 2·6 Twenty-eight-dayrepeated-dose oral toxicity study in rats Bll-0908 Detailed clinical observations ofindividual animals (week2)

Exp.group Removal fromcage Animal No. (mglkglday) Ease ofremoval Vocalization 31 0 0 

### 32 0 0 

### 33 0 0 34 0 0 Vehicle 35 0 0 control 36 0 0 37 0 0 38 0 0 39 0 0 40 0 0 41 0 0 42 0 0 25 43 0 0 44 0 

### 0
Female 45 0 0 46 0 0 47 0 0 150 48 0 0 49 0 0 50 0 0 51 0 0 52 0 0 53 0 0 54 0 0 55 0 0 1,000 56 0 0 57 0 0 

### 58 0 0 59 0 0 60 0 +l



## -100­ 366Addendum 2·7 Twenty-eight-day repeated-dose oral toxicity study in rats BI1-0908 Detailedclinical observations ofindividual animals (week 3)

Exp.group Removal from cage Sex AnimalNo. (mglkglday) Ease ofremoval Vocalization 0 0 

### 2 0 0 

### 3 0 0 4 0 0 Vehicle 5 0 0 control 

### 6 0 0 7 0 0 8 0 0 9 0 0 10 0 0 11 0 0 12 0 0 25 13 0 0 14 0 0 

### Male IS 0 0 16 0 0 17 0 0 150 18 0 0 19 0 0 20 0 0 21 0 0 22 0 0 23 0 0 24 0 0 1,000 25 

### 26 0 0 0 0 

### 27 0 0 

### 28 0 +1 

### 29 0 0 

### 30 0 0



## -101­ 367Addendum 2·8 Twenty·eight-dayrepeated-dose oral toxicity studyin rats B1l..()9{)8 Detailed clinical observations ofindividual animals (week3)

Exp.group Removal from cage Animal No. (mglkg/day) Ease ofremoval Vocalization 31 0 0 32 0 0 33 0 0 34 0 0 Vehicle 35 0 0 control 36 0 0 37 0 0 38 0 0 39 0 0 40 0 0 41 -I 0 42 0 0 25 

### 43 0 0 44 0 0
female 45 0 +1 46 0 +1 47 0 0 

### ISO 48 0 0 

### 49 0 0 50 0 0 51 0 0 52 0 0 53 0 0 54 0 0 1,000 55 56 0 0 +l 0 57 0 0 58 0 0 59 0 0 60 0 0



## -102­ 368Addendum 2-9 Twenty-eigbt-day repeated-doseoral toxicity studyin rats Bll·0908 Detailed cUnical observations ofindividual animals (week4)

Exp.group Removal from cage 

### Sex AnimalNo. (mglkglday) Ease of removal Vocalization 0 0 2 0 0 3 0 0 4 0 0 Vehicle 5 0 0 control 6 0 0 7 0 

### +1 8 0 0 9 0 0 10 0 0 11 0 

### +1 12 ·1 0 25 13 0 0 

### 14 0 0 Male 15 0 0 16 0 0 17 0 0 150 

### ]8 0 0 

### 19 0 

### +1 20 0 0 21 0 0 22 0 0 23 0 

### +1 24 0 0 1,000 2S 26 0 

### 0 0 0 

### 27 0 0 

### 28 0 

### 0 

### 29 0 0 30 0 

### +]



## -103­ 369Addendum 2-10 Twenty-eight-day repeated-dose oral toxicity study in rats B11-0908 Detailed clinical observations ofindividual animals (week

### 4)

Exp.group Removal from cage Sex AnimalNo. (mg/kgfday) Easeofremoval Vocalization 

### 31 0 0 

### 32 0 

### 0 33 

### 0 +) 

### 34 0 0 Vehicle 

### 35 

### 0 0 control 36 

### 0 0 37 0 0 

### 38 0 

### +1 

### 39 

### 0 0 

### 40 0 +1 41 

### 0 +1 42 

### 0 +\ 25 

### 43 0 

### 0 44 0 

### 0
Female 

### 45 0 

### 0 46 0 

### 0 47 0 0 150 

### 48 

### 0 0 49 

### 0 0 50 0 0 

### 51 

### 0 0 

### 52 0 0 

### 53 0 +1 54 -\ 0 1,000 

### 55 

### 56 0 0 0 

### +\ 

### 57 0 0 

### 58 0 0 

### 59 

### 0 0 

### 60 0 

### 0



## -104­ 370Addendum 2-11 Twenty-eight-day repeated-dose oral toxicity study in rats BIl-0908 Detailed clinical observations ofindividual animals (Recovery week I)

Exp.group Removal from cage Sex AnimalNo. (mglkglday) Ease

# ofremoval Vocalization 

### 6 0 0
Vehicle control 

### 7 8 9 0 0 0 0 0 0
Male 10 

### 26 0 0 0 0 

### 27 0 

### +) 1,000 

### 28 0 

### +1 29 0 0 30 0 

### +)



## -105­ 311Addendum 2-12 Twenty-eight-day repeated-dose oral toxicity study in rats Bll-0908 Detailed clinical observations ofindividual animals (Recovery week 1)

Exp.group Removal from cage Selt AnimalNo. (mglkg/day) Ease ofremoval Vocaliwion 36 0 0 

### 37 0 

### 0 Vehicle control 

### 38 0 0 

### 39 0 0
Female 

### 40 0 0 

### 56 0 0 

### 57 0 0 1,000 

### 58 0 0 .59 0 0 

### 60 0 0



## -106­ 372Addendum 2-J3 Twenty-eight-day repeated-dose oral toxicity study in**rat~** BII-0908 Detailed clinical observations ofindividual animals (Recovery week 2)
Sex 

### Exp.group (mglkg/day) AnimalNo. Removal Ease ofremoval from cage

6 0 0
Vehicle control 7 8 

### 9 0 0 0 +1 

### 0 0
Male 10 

### 26 0 0 

### 0 0

1,000 

### 27 28 29 0 0 0 0 0 0 

### 30 0 0 VoealiZlltion



## -107­ 

## 373Addendum 2-14 Twenty-eight-dayrepeated-dose oral toxicity study in rats B1I·0908 Detailed clinical observations ofindividual animals (Recovery week 2)

Exp.group Removal from cage Sex AnimalNo. (mglkglday) Easeofremoval Vocalization 36 0 0 37 0 0 Vehicle control 38 0 0 39 0 0 

### Female 40 0 +1 

### 56 0 0 57 0 +1 1,000 

### 58 0 

# ° S9 

# ° 0 60 

# ° 0



## -108­ 374Addendum 2·15 Twenty-eight-day repeated·dose oral toxicity study in rats Bll·0908 Detailed clinical observations ofindividual animals (Predosing)Handling observations 

### Sex I::':xp.group Animal No. Muscle Subnonnal Piloerection Staining Unkempt Paleness Reddening (mg/kglday) tone temperature hair hair 0 2 () 3 0 4 0 Vehicle 5 0 control 

### 6 0 7 0 

### 8 0 9 0 10 0 11 0 12 0 

### 2S 13 0 14 0
Male 

### lS 0 

### 16 0 17 0 150 18 0 

### 19 0 

### 20 0 21 0 22 0 23 0 24 0 25 0 1,000 26 0 27 0 

### 28 0 29 0 30 0



## -109­ 

## 375911-0908 Addendum 2-16 Twenty-eight-dayrepeated-dose oral toxicity study in rats Detailed clinical observations ofindividual animals (Predosing)Handling observations Sex Exp. group Animal No. Muscle Subnormal Piloerection Staining Unkempt Paleness Reddening (mglkgfday) tone temperature hair hair 31 0 32 0 33 0 34 0 Vehicle 35 0 control 36 0 37 0 38 0 39 0 40 0 41 0 42 0 25 43 0 44 0
Female 45 0 46 0 47 0 150 48 0 49 0 50 0 51 0 52 0 53 0 54 0 1,000 55 0 56 0 57 0 58 0 59 0 

### 60 0



## -110­ 376Addendum 2-17 Twenty-eight-day repeated·dose oral toxicity study in**ra~** 

### B11-0908 Detailed clinical observations ofindividual animals (week 1) Handling observations Sex 

### Exp. group Animal No. Muscle Subnormal Piloerection Staining Unkempt Paleness Reddening (mglkglday) tone 

### temperature hair hair 1 0 2 0 3 0 4 0 

### Vehicle 5 0 control 6 0 7 0 

### 8 0 9 0 10 0 11 0

25 12 13 0 0

Male

ISO 14 IS 16 17 18 0 0 0 0 0 19 0 20 0 21 0 22 0 23 0 24 0 1,000 25 26 0 0 27 0 28 0 29 0 30 0



### -111­ 377Addendum 2-18 Twenty-eight-day repeated-dose oral toxicitystudy in rats BI1-0908 Detailed clinical observations ofindividual animals (week 1) Handling observations Sex Exp. group AnimalNo. Muscle Subnormal 

### Piloerection Staining Unkempt Paleness Reddening (mglkglday) tone 

### temperature hair hair 31 0 

### 32 0 

### 33 0 

### 34 0 Vehicle 

### 35 0 control 36 0 

### 37 0 38 0 39 0 

### 40 0 

### 41 0 42 0 

### 25 

### 43 0 

### 44 0 Female 45 0 

### 46 0 

### 47 0 

### ISO 48 0 49 

### 0 50 0 51 0 52 0 53 0 +") 54 0 

### 1,000 

### 5S 56 0 0 +1) 

### 57 0 

### 58 0 59 0 + .) 60 

### 0 a) Lower abdomen



## -112­ 378Addendum 2·19 Twenty-eight-day repeated-dose oral toxicity study in rats B11·0908 Detailed clinical observationsofindividual animals (week2) Handlingobservations Sex 

### Exp. group Animal No. Muscle SubnofTIUIl Piloerection Staining Unkempt Paleness Reddening (mglkglday) tone temperature hair hair 1 0 2 0 3 0 4 0 Vehicle 

### 5 0 control 

### 6 0 7 0 8 0 9 0 10 0



### 25 II 12 13 14 0 0 0 0 Male IS 16 0 0

ISO 17 18 19 0 0 0 20 0

],000 21 22 23 24 

### 25 26 27 28 29 30 0 0 {) 0 0 0 0 0 0 0



## -113­ 379Addendum 2·20 Twenty-eight-day repeated-dose oral toxicity study in rats BII·0908 Detailed clinical observations ofindividual animals (week 2) Handling observations

Exp. group AnimalNo. 

### Muscle Subnormal Piloerection Staining Unkempt Paleness Reddening (mg/kglday) tone temperature hair hair 

### 31 

### 0 32 0 33 

### 0 34 0 Vehicle 

### 35 0 control 36 0 

### 37 0 38 0 39 0 

### 40 0 41 0 42 

### 0 25 43 0 44 0 Female 

### 45 0 

### 46 0 

### 47 0 

### 150 48 0 49 0 50 0 51 0 52 

### 0 53 0 54 0 1,000 55 56 0 0 

### 57 0 +") 

### 58 0 

### 59 0 

### 60 0 II)Lower abdomen



## -114­ 380Addendum 2-21 Twenty-eigbt-day repeated-dose oral toxicity study in rats 811-0908 Detailed clinical observationsofindividual animals (week 3) Handling observations 

### Sex Exp. group Animal No. Muscle Subnonnal Piloerection Staining Unkempt Paleness Reddening (mglkgfday) tone temperature hair hair 0 2 0 3 0 4 0 Vehicle 5 0 control 6 0 

### 1 0 8 0 9 0 10 0 11 0

Male 

### 25

150 12 13 14 15 16 

### 11 18 0 0 0 0 0 0 0 19 

### 20 0 0



### 1,000 

### 21 22 23 24 

### 2S 

### 26 21 

### 28 29 30 0 0 0 0 0 0 0 0 0 0



## -115­ 381Addendum 2-22 Twenty-eight-day·repeated-dose oral toxlcity study in rats 811-0908 Detailed clinical observations ofindividual animals (week 3) Handling observations Sex 

### Exp. group AnimalNo. 

### Muscle SubnonnaJ Piloerection Staining 

### Uulcempt Paleness ReddeI)ing (mg/kglday) tone temperature hair hair 

### 31 0 32 0 33 0 34 0 Vehicle 

### 3S 0 control 36 0 37 0 

### 38 0 39 0 40 0 41 0


### 2S 42 43 0 0 44 0 

### Female 

### 4S 46 0 0

ISO 47 48 0 0 49 0 50 0 51 0 52 0 53 0 54 0 1,000 

### SS 56 0 0 +a)

57 0 58 0 59 0 60 0 a) Lowerabdomen



## -116­ 382Addendum 2-23 Twenty-eighl-dayrepealed.dose oral toxicity study in rats 

### BJ 1·0908 Detailed clinical observations ofindividual animals (week4) Handling observations Sex 

### Exp. group Animal No. Muscle Subnormal Piloerection Staining Unkempt Paleness Reddening (mglkglday) 

### tone temperature hair hair I 0 2 0 3 0 4 

### 0 Vehicle 5 0 control 6 0 7 0 8 0 9 0 10 0 11 0 12 0 

### 25 13 0 14 0
MaJe 

### 15 0 

### 16 0 17 0 

### 150 18 0 19 0 20 0 

### 21 0 22 0 +") 

### 23 0 

### 24 0 

### 25 0 1,000 

### 26 0 

### 27 0 +") 28 0 +t) 29 0 +h) 30 0 a) Moistening ofthemandibule andneck: b) Moistening ofthemandibule



## -117­ 383Addendum 2·24 Twenty-eight-day repealed·dose oral toxicity studyin rats BII-0908 Detailedclinical observationsofindividual animals <week4) Handlingobservations Sex Exp. group Animal No. Muscle Subnonnal Piloerection Staining Unkempt 

### Paleness Reddening (mg/lcglday) tone 

### temperature hair bair 31 

### 0 32 

### 0 33 

### 0 34 0 Vehicle 35 0 control 36 0 37 0 38 0 3!) 0 40 0

25 41 42 43 0 0 0 44 0 Female 45 46 0 0

150 47 48 4!) .50 0 0 0 0 51 0 52 0 + oj 53 0 +a) 54 0 

### 1,000 55 56 0 

### 0 +a)

+ &),0) 57 0 + oj 58 0 + 0) 59 0 60 0 +a) a) Lowerabdomen b)Moisteningoftbemandibule



## -118­ 384Addendum 2-25 Twenty-eight-day repeated-dose oral toxicity study in rats Bll-0908 Detailed clinical observations ofindividual animals (Recovery week 1) Handling observations Sex Exp.group Animal No. Muscle Subnormal Piloerection Staining Unkempt Paleness Reddening (mglkglday) tone temperature hair hair 6 0 7 0 Vehicle control 8 0 9 0
Male 10 0 26 0 27 0 1,000 28 0 29 0 30 0



## -119­ 385Addendum 2·26 Twenty-eight·day repeated·dose oraltoxicity study in rats .B11-0908 Detailed clinical observations ofindividual animals (Recovery week 1) Handling observations Exp. group AnimalNo. Muscle Subnormal Piloerection Staining Unkempt Palel'les5 Reddening (mglkglday) tone temperature hair hair 36 0
Vehicle control 37 38 39 0 0 I)
Female 40 S6 0 0 57 0 1,000 

### 58 0 59 0 60 0



## -120­ 386Addendum 2·27 Twenty·eight-dayrepeated-dose oral toxicity study in rats BlI·Ol108 Detailed clinical observations ofindividualanimals (Recovery week2) Handling observations Exp.group (mglkgfday) Animal No. Muscle tone Subnormal temperature Piloerection Staining hair Unkempt hair 

### Paleness Reddening

6 0
Vehicle control 7 8 9 0 0 0
Male 10 26 t} 0**.~** 27 0 1,000 28 0 29 0 

### 30 0



## -121­ 387Addendum2·28 Twenty-eight-day repeated·dose oral toxicity studyin rats B11-0908 Detailed clinical observations ofindividual animals (Reroveryweek2) Handling observations Exp. group AnimalNo. Muscle Subnonnal Piloerection Staining Unkempt Paleness Reddening 

### (mglkJldal') tone temperature 

### bair hair 36 0 37 0 Vehicle control 38 0 

### 39 0
Female 40 0 56 0 57 0 1,000 58 0 59 0 

### 60 0



## -122­ 388Addendum 2-29 Twenty-eight-day repeated-dose orol toxicity study in rats 811-0908 Detailed clinical observations ofindividual animals (predosing) Handling observations Sex Exp.group Animal No. Cyanosis Lacrimation Exophthalmos Pupillary Salivation Secretion (mglkglday) 

### sill: 0 2 0 3 0 4 0 Vehicle 5 0 control 

### 6 0 7 0 

### 8 0 9 0 10 0 11 0 12 0 25 13 0 14 0 15 0 Male 

### 16 0 17 0 150 

### 18 0 

### 19 

## a 20 0 21 0 22 0 23 0 24 0 1,000 25 

### 26 0 0 

### 27 0 28 0 

### 29 

### 0 30 

### 0



## -123­ 389Addendum 2-30 Twenty-eight-day repeated-doseoral toxicity study inrats BIJ-0908 Detailedclinical observations ofindlvidual animals (Predosing) Handlingobservations Sex Exp. group Animal

## No. Cyanosis Lacrimation Exophthalmos Pupillary Salivation Secretion (mglkglday) size 31 0 32 0 33 0 34 0 Vehicle 35 0 control 36 0 37 

### 0 38 0 39 

### 0 40 0 41 0

Female 25 42 43 44 45 46 0 0 0 

### 0 0

150 47 48 

### 49 50 0 0 0 0 51 0 52 0 53 

# ° 54 

# ° S5 1,000 

# ° 56 0 57 0 58 0 59 0 60 0



# -124­ 390Addendum 2-31 Twenty-eight.day repeated.dose oral toxicity study in rats BlI-0908 Detailed clinical observations ofindividual animals (week J) Handlingobservations Sex. Exp. group Animal No. Cyanosis Lacrimation Exophthalmos Pupillary 

### Salivation Secretion (mglkglday) size 0 2 0 3 0 4 0 Vehicle 5 0 control 6 0 

### 7 0 II 0 9 0 10 0

25 II 12 13 14 0 0 0 0
Male

150 15 16 17 18 19 20 0 0 0 0 0 0 21 0 

### 22 0 23 0 24 0 l,Ooo 25 

### 26 0 0 

### 27 0 28 0 

### 29 

# ° 30 0



## -125­ 391Addendum2-32 Twenty-eight-day repeated-dose oral toxicity study in rats 

### Bll-0908 Detailed clinical observations ofindividual animals (week I) Handling observations Sex Exp. group AnimalNo. Cyanosis Lacrimation Exophthalmos Pupillary Salivation Secretion (mglkglday) size 31 0 32 0 33 0 34 

# - 0 Vehicle 

### 35 0 control 

### 36 0 

### 37 0 

### 38 0 

### 39 0 

### 40 0 

### 41 0 

### 42 0 

### 25 43 0 

### 44 0 Female 

### 45 0 

### 46 0 

### 47 0 

### 150 

### 48 0 

### 49 0 

### SO 0 51 0 

### 52 0 

### 53 0 

### 54 0 1,000 55 

### 56 0 0 

### 57 0 

### 58 0 59 0 60 

### 0



## -126­ 392Addendum 2·33 Twenty·eight-day repeated-dose oral toxicity study in ralS B11·0908 Detailed clinical observations ofindividual animals (week2) Handling observations Sex Exp.group AnImll.INo. Cyanosis Lacrimation Exophthalmos Pupillary Salivation Secretion (mglkglday) size 0 2 0 3 0 4 0 Vehicle 5 0 control 6 0 7 0 8 0 9 0 

### 10 0 11 0 12 0 25 13 0 14 0 Male IS 0 16 0 17 0 150 18 0 19 0 20 0 

### 21 0 

### 22 0 

### 23 0 24 0 1,000 

### 25 

### 26 0 0 27 0 

### 23 0 

### 29 0 30 0



## -127­ 393Addendum 2-34 Twenty-eight-day repeated-dose oral toxicity study in rats B11-0908 Detailed clinical observations ofindividual animals (week2) Handling observations Sex Exp.group Animal No. Cyanosis Lacrimation Exophthalmos Pupillary Salivation Secretion (mglkgfday) size 31 0 32 0 33 0 34 

### Q Vehicle 35 0 control 36 0 37 0 38 0 39 0 40 0 41 0 42 0 25 43 0 44 0
Female 45 0 46 0 47 0 150 48 0 49 0 50 0 51 0 52 0 53 0 54 ·0 1,000 55 0 56 0 57 0 58 0 59 0 60 0



## -128­ 394Addendum 2-35 Twenty-eight-dayrepeated-dose oraltoxicity study in rats BlI-0908 Detailed clinical observations ofindividual animals (week 3) Handling observations 

### Sex Exp. group Animal No. Cyanosis Lacrimation Exophthalmos Pupillary 

### Salivation Secretion (mglkg/day) size 0 2 0 3 0 

### 4 0 Vehicle 5 0 control 6 0 7 0 8 0 9 0 10 0 11 0 12 0 2.5 13 0 14 0 Male IS 0 16 0 17 0 150 18 0 19 0 

### 20 0 21 0 22 + 0 23 

### 0 

### 24 0 1,000 

### 2.5 

### 26 0 0 27 0 28 0 29 0 30 0



## -129­ 395Addendum 2-36 Twenty-eight-day repeated-doseoralloxicity study in rats 

### 811-0908 Detailed clinical observetions ofindividualanimais (week 3) Handling observations Sex Exp. group AnimalNo. Cyanosis Lacrimation Exophthalmos Pupillary Salivation Secretion (mgllcglday) 

### size 31 0 32 

### 0 33 

### 0 34 0 Vehicle 

### 35 0 control 

### 36 0 37 0 38 0 39 0 

### 40 0 41 

### 0 42 0 2:5 43 0 44 0 Female 

### 45 46 0 0



### 150 47 48 

### 49 0 0 0 

### 50 0 

### 51 0 S2 0 

### 53 0 

### 54 0 1,000 

### 55 56 0 0 

### 57 0 

### 58 0 

### 59 0 60 0



## -130­ 396Addendum 2-37 TWCllty-cigbt-dayrepeated-dose oral tox,icity study 

### in rats B11-0908 Detailed clinical observations ofindividual animals (week4) Handling observations Sex Exp. group Animal No. Cyanosis Lacrimation Ex,ophthalmos Pupillary Salivation Secretion**(m~day)** 

### size 0 2 

### 0 3 0 4 0 Vehicle 5 0 

### control 6 0 7 0 8 

### 0 9 0 10 0 11 0 12 0 25 

### 13 0 14 0
Male 15 0 16 0 17 0 150 18 0 

### 19 0 20 0 21 0 22 

### + 0 23 0 24 0 25 0 1,000 26 0 27 0 28 0 

### 29 0 30 0



# -131­ 397Addendum 2-38 Twenty-eight-dayrepeated-dose oral toxicity studyin rats BI1-0908 Detailed clinical observations ofindividual animals (week 4) Handlingobservations Sex Exp. group Animal No. Cyanosis Lacrimation Exophthalmos Pupillwy Salivation Secretion (mg/kg/day) size 31 0 32 0 33 0 34 0 Vehicle 35 0 control 36 0 37 0 38 0 39 0 40 0 41 0 42 0 25 43 0 44 0 45 0 Female 46 0 47 0 150 48 0 49 0 50 0 51 0 52 0 53 0 54 0 1,000 55 0 56 0 57 0 58 0 59 0 60 0



## -132­ 398Addendum 2-39 Twenty-eight-day repeated-dose oral toxicity study in rats 811-0908 Detailed clinical observations ofindividualanimals(Recovery week I) Handlingobservations Sex Exp.group Animal No. Cyanosis Lacrimation Exophthalmos Pupillary Salivation Secretion (mgfkglday) size 6 0 7 0 Vehicle control 8 0 9 Q 10 0 Male 

### 26 0 27 0 1,000 28 0 29 0 30 0



## -133­ 399Addendum 2-40 Twenty-eight-dayrepeated-dose oral toxicitystudy in rats BII.-0908 Detailed clinical observations ofindividual animals (Recovery week: 1) Handling observations Sex Ex.p. group Animal No. Cyanosis Lacrimation Exophthalmos Pupillary Salivation Secretion (mglkg/day) size 36 

### o 37 

### o Vehicle control 38 

## o 39 

### o
Female 

### 40 

## o 56 

### o 57 

### o J,OOO 58 

## o 59 

## o 60 

## o



## 134­ 400Addendum 2-41 Twenty-eight-day repeated-dose oral toxicity studyinrats B11-0908 Detailed clinical observations ofindividual animals (Recovery week2) Handling observations Exp.group 

### AnimalNo. Cyanosis Lacrimation Exophthalmos Pupillary Salivation 

### Secretion (mglkgfday) size 

### 6 0 

### 7 0 Vehicle control 8 0 

### 9 0
Male 10 0 

### 26 0 

### 27 0 1,000 

### 28 0 

### 29 0 30 0



## -135­ 401Addendum 2-42 Twenty-eight-day repeatcd-dosc oral toxicity study in rats BIl-0908 Detailed clinical ooservations ofindividual animals (Recovery week2) Handlingobservations 

### Sex Exp.group Animal No. Cyanosis Lacrimation Exophtbalmos Pupillary Salivation Secretion (rnglkglday) size 36 0 37 0 Vehicle control 38 0 

### 39 0 Female 40 0 56 0 57 0 1,000 58 0 

### 59 0 60 0



## -136­ 402Addendum 2-43 Twenty·eight-day repea1ecl-dose oral toxicity study in rats Bll-0908 Detailed clinical observations ofindividual animals (Preclosing) Observations in arena Sill( Exp.group AnimalNo. Posture Motor Respiration Lid 

### Gail (mglkglday) activity closure 1 0 0 0 2 0 0 0 3 0 0 

### 0 4 0 0 0 Vehicle 

### S 0 0 0 control 6 0 0 0 7 0 

### 0 0 8 0 0 0 9 0 0 0 10 0 +1 0 11 0 0 0 12 0 0 0 25 13 0 0 0 14 0 0 0 

### Male IS 0 

### 0 0 16 0 0 0 17 0 0 0 

### ISO 18 0 0 

### 0 J9 0 0 0 

### 20 0 0 

### 0 21 0 0 0 

### 22 0 0 0 

### 23 0 0 0 

### 24 0 0 0 1,000 

### 25 26 0 0 0 

### 0 0 0 27 0 0 0 

### 28 0 0 0 

### 29 0 0 0 

### 30 0 0 

### 0



## -137­ 403Addendum 2-44 Twenty-eight·dayrepeated-dose oral toxicity study in rats Bll·0908 Detailed clinical observations ofindividual animals (Preclosing) Observations in arena Sex Exp. group AnirnalNo. Posture Motor Respiration 

### Lid Gait (mglkglday) activity closure 31 0 0 0 32 0 0 0 33 0 0 0 

### 34 0 0 0 Vehicle 35 0 Q 0 control 36 0 0 0 37 0 0 0 38 0 0 0 39 0 +1 0 

### 40 0 0 

### 0 41 0 0 0 42 0 0 0 25 

### 43 0 0 0 44 0 0 0 

### Female 45 0 0 0 

### 46 0 0 0 47 0 0 0 150 

### 48 0 0 0 

### 49 0 0 0 50 0 

# ° 0 51 0 0 0 52 0 0 0 53 0 

### 0 0 

### 54 0 

### 0 0 1,000 

### 55 56 0 0 

### 0 0 0 0 57 0 0 0 58 0 0 0 59 

# ° 0 0 60 

# ° 0 0



## -138-­ 404Addendum 2-45 Twenty-eight-dIlYrepeated-dose oral toxicity study in rats Bll-0908 Detailed clinical observations ofindividual Mimals (week 1) Observations in arena Sex Exp. group AnimalNo. Posture Motor Respiration Lid Gait (mglkgfday) activity closure 0 0 0 2 0 -1 0 

### 3 0 0 0 4 0 0 0 Vehicle 5 0 0 0 control 6 0 0 0 7 0 0 0 8 0 +} 0 9 0 0 0 10 0 +1 0 II 0 0 0 12 0 0 0 25 \3 0 0 0 14 0 0 0 Male 15 0 +1 0 16 0 0 0 17 0 0 0 150 18 0 +1 

### 0 19 0 0 0 20 0 

### 0 0 21 0 0 0 22 0 0 0 23 0 0 0 24 0 0 0 1,000 25 26 0 0 

### 0 0 0 0 27 0 0 0 28 0 0 0 29 0 +1 0 

### 30 0 0 0



## -'-139­ 405Addendum 2-46 Twenty-eight-dayrepeated-dose oral toxicitystudy in rats 911-0908 Detailed clinical observations ofindividual animals (week I) Observations in arena Sex. Exp. group Animal No. Posture Motor Respiration Lid Gait (mglkgJday) aetlvity closure 31 0 0 0 32 0 +l 0 33 0 0 0 34 0 0 0 Vehicle 35 0 0 0 control 36 0 +1 0 37 0 0 0 38 0 0 0 39 0 0 0 40 0 +1 0 41 0 0 0


### 2S 42 43 0 0 0 0 0 0

Female

150 44 45 46 47 48 49 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 50 0 0 0 51 0 0 0 52 0 0 0 53 0 0 0 54 0 0 0 1,000 

### 55 56 0 0 0 +1 0 0 57 0 0 0 58 0 +1 0 59 0 0 0 60 0 0 0



## -140­ 406Addendum 2-47 Twenty-eight-da.yrepeated-dose oral toxicity study in rats BII-0908 Detailed clinical observationsofindividuaillnimals (week2) Observationsinarena Sex Exp.group Animal No. Posture Motor Respiration Lid Gait (mglkglday) activity closure 

### 0 Q 0 2 0 0 0 3 0 0 0 4 0 0 0 Vehicle 5 0 0 0 

### control 6 0 0 0 7 0 0 0 8 0 0 0 9 0 Q 0 10 Q 

### +1 0 11 0 0 0 

### 12 0 0 0 

### 25 13 0 0 0 14 0 

### 0 0
Male 15 

### 0 0 Q 16 0 0 0 17 

### 0 0 0 150 18 0 0 0 19 0 0 0 

### 20 0 0 0 21 0 0 0 

### 22 0 0 0 23 

### 0 0 0 24 0 0 0 1,000 25 

### 26 Q 0 0 

### +1 Q 0 27 0 0 0 

### 28 0 0 

### 0 

### 29 0 +1 0 30 0 0 0



## -141­ 407Addendum 2-48 Twenty-eight-day repeated-dose oral toxicity study in rats BII-0908 Detailed clinical observations ofindividual animals (week2) Observations in lU'eTla 

### Sex Ex.p. group AnimalNo. Posture Motor RespinUion Lid Gait (mgIkg{day) activity closure 3] 0 0 

### 0 32 0 0 0 33 0 0 0 34 0 0 .0 Vehicle 35 0 0 0 control 36 0 0 0 

### 37 0 0 0 38 0 0 0 39 0 0 0 40 0 0 0 41 0 0 0 42 0 0 0 25 43 0 0 

### 0 44 0 

### 0 0 Female 45 0 +l 0 46 0 0 0 47 0 0 0 150 48 0 0 0 49 0 0 0 

### SO 0 0 0 51 0 0 0 52 0 0 0 53 0 0 0 54 0 0 0 

### 1,000 55 56 0 0 0 0 0 0 57 

### 0 0 0 58 0 0 0 59 0 0 0 60 0 +1 0



## -142­ 408Addendum 2-49 Twenty-eight-day repeated-dose oral toxicity study in rats Bll-0908 Detailed clinical obse("\lations ofindividual animals (week 3) Ob5el"\iations in arena Sex Exp. group Animal No. Posture Motor Respiration Lid Gait (mglkglday) activity closure I 0 0 0 2 0 0 0 3 0 0 0 4 0 0 0 :' Vehicle 5 0 0 0 control 6 0 0 0 7 0 0 0 8 0 0 0 

### 9 0 0 0 10 0 0 0 11 0 0 0 12 0 0 0 25 13 0 0 0 

### ]4 0 0 0
Male 15 0 0 0 16 0 0 0 17 0 0 0 150 18 0 0 0 19 0 0 0 

### 20 0 0 0 21 0 0 

### 0 22 0 0 0 23 0 0 0 

### 24 0 0 0 1,000 

### 2S 

### 26 0 0 0 0 0 0 27 0 

### 0 0 28 0 0 0 

### 29 0 

### 0 0 30 0 0 0



## -143­ 409Addendum 2·50 Twenty-eight-dayrepeated-dose oral toxicity study in rats BI1·0908 Detailed clinical observations ofindividual animals(week: 3) Observations in arena Sex Exp. group AnimalNo. Posture Motor Respiration Lid Galt (mglkglday) activity closure 31 0 0 0 32 0 0 0 33 0 0 0 34 0 0 0 Vehicle: 35 0 0 0 control 36 0 0 0 37 0 0 0 38 0 0 0 39 0 0 0 40 0 0 0 41 0 0 0 42 0 0 0 25 43 0 0 0 44 0 0 0
Female 45 0 0 0 46 0 0 0 47 0 0 0 150 48 0 0 0 49 0 0 0 50 0 0 0 51 0 0 0 52 0 0 0 53 0 0 0 .54 0 0 0 

### 1,000 55 .56 0 0 0 0 0 0 57 0 0 0 58 0 0 0 59 0 0 0 60 0 0 0



# -144­ 410Addendum 2-51 Twenty-eight-dayrepeated·dose oral toxicity study in rats 811...0908 Detailed clinical observations ofindividual animals (week4) Observations in arena Sex 

### Exp. group Animal No. Posture Motor Respiration Lid 

### Oait (mgtkglday) activity closure 0 0 0 2 

### 0 -1 0 3 0 0 0 

### 4. 0 0 0 Vehicle 5 0 0 0 control 6 0 0 0 7 0 0 0 8 0 0 0 9 0 0 0 10 0 0 0 11 0 0 0 12 0 0 0 25 13 0 0 0 14 0 0 0
Male 15 0 

### -I 0 16 0 0 0 17 0 0 0 150 18 0 0 0 19 0 +1 

### 0 20 0 0 0 21 0 

### 0 0 22 0 -I 0 23 0 0 0 24 0 0 0 1,000 25 26 0 0 0 0 0 0 27 0 -I 0 28 0 

### 0 0 29 0 0 0 30 0 0 0



## -145­ 411Addendum 2·52 Twenty-eight-dayrepeated-dose oral toxieity study in rats Bll-0908 Detailed clinical observations ofindividual animals (week4) Observations in arena Sex 

### Exp. group Animal No. Posture Motor Respiration Lid 

### Gait (mgllc&lday) 

### activity closure 

### 31 0 0 0 32 0 +1 0 

### 33 0 0 0 

### 34 0 0 0 Vehicle 35 0 0 0 conlrol 

### 36 0 0 0 37 0 0 0 

### 38 0 0 0 39 0 0 0 40 0 -I 0 41 0 0 0 

### 42 0 0 0 25 

### 43 0 0 0 44 0 0 0 Female 

### 45 0 0 0 46 0 0 0 

### 47 0 0 0 150 

### 48 0 0 0 

### 49 0 0 0 

### SO 0 0 0 51 0 0 0 52 0 0 0 53 0 0 0 54 0 -1 0 1,000 

### S5 56 0 0 +1 +I 0 0 57 0 +1 0 58 0 0 0 

### 59 0 0 0 60 0 0 0



## -146­ 412Addendum 2-53 Twcnty-eight-day repeated-dose oral toxicity study in rats Bl1-0908 Detailed clinical observations ofindividual animals (Recovery week I) Observations in arena Sex Exp.group Animal No. Posture Motor Respiration Lid Galt (mglkg/day) activity closUre 6 

# ° 0 0 7 0 -1 0 Vehicle control 8 0 0 0 9 0 0 

# ° Male 10 0 0 

# ° 26 0 

# ° ° 27 0 0 0 1,000 28 

# ° ° ° 29 0 

# ° 0 30 0 0 0



## -147­ 413Addendum 2·54 Twenty-eight·day repeated.dose oral toxicity study inrats B11-0908 Detailed clinical observations ofindividual animals (Recovery week 1) Observations in arena Sex Exp. group AnimalNo. Posture Motor Respiration Lid Gait (mg1kglday) activity closure 36 0 0 0 37 0 0 0 Vehicle control 38 0 0 0 39 0 0 0 Female 

### 40 0 0 

### 0 56 

# ° 0 

# ° 57 0 0 0 1,000 58 

# ° 0 0 59 0 0 0 60 0 0 0



## -148­ 414Addendum 2-55 Twenty-eight-day repeated-dose DraltDxicity study in rrlts B11-0908 Detailed clinical observations ofindividual animals (Recovery week2) Observations in arena Sex Exp.group Animal No. Posture Motor Respiration Lid Gail (mglkglday) activity closure 

### 6 0 0 0 7 0 -1 0 Vehicle cantrol 8 0 0 0 9 0 0 0 Male 10 

# ° 0 0 

### 26 0 0 

# ° 27 0 0 0 1,000 28 0 0 0 29 0 0 0 30 

### 0 0 0



## -149­ 415Addendum2·56 Twenty-tight-dayrepeated·dose oral toxicity study in rats Bll·0908 Detailed clinical observations ofindividual animals (Recovery week 2) Observations in arena Sex Exp. group AnimalNo. Posture Motor Respiration Lid Gait (mgfkglday) activity closure 36 0 +1 0 37 0 0 0 Vehicle control 38 0 0 0 3C;? 0 0 0
Female 40 0 0 0 56 0 0 0 57 0 

# ° ° 1,000 

### 58 0 

# ° 0 59 0 0 0 60 

# ° 0 

# °

-150­ 416Addendum 2-57 Twenty-eight-dayrepeated-dose oral toxicity study in rats BII·0908 Detailed clinical observations ofindividual animals (Prcdosing) Observations in arena Sex Exp.group AnimalNo. Tremoritwitchl Defecation Urination Stereotypic Abnormal (mglkglday) convulsion (count/min) (count/min) behavior behavior 0 0 3 2 0 0 0 3 0 1 2 4 0 0 3 Vehicle 5 0 0 0 control 6 0 0 0 7 0 11 8 0 0 9 0 0 0 10 0 0 I 11 0 0 0


### 25 12 13 0 0 0 0 0 21 14 0 0 0 Male 15 16 0 0 0 0 J



### ISO 17 18 0 0 0 0 16 

### 3 J9 0 0 0 20 0 2 21 0 0 I 22 0 0 0 

### 23 0 0 

### 2 24 0 0 S 1,000 

### 25 0 0 0 26 0 0 I 27 0 0 

### 2 28 0 0 0 29 0 1 

### 3 30 0 0



## -151­ 417Addendum 2-58 Twenty-eight-dayrepeated-dose oral toxicity study in rats Bl1-0908 Detailed clinical observations ofindividual animals (Predosing) Observations in arena Sex Exp. group (mglkglday) AnimalNo. Tremor/twitch! convulsion Defecation (countlmin) Urination (count/min) Stereotypic behavior Abnormal behavior 31 0 0 0 

### 32 0 0 I 

### 33 0 0 0 34 0 0, 4 Vehicle 

### 35 0 

### 0 

### 3 control 

### 36 0 

### 0 5 37 0 0 4 38 0 

### 0 39 0 0 2 40 0 0 0 41 0 0 0 42 0 0 0 25 43 0 0 7 44 0 0 13 Female 45 0 0 2 46 0 0 2 47 0 0 9 150 48 0 0 0 

### 49 0 0 0 SO 0 I 0 51 0 0 4 52 0 0 0 53 0 0 0 S4 0 0 0 \,000 

### 55 0 0 3 56 0 1 0 57 0 0 3 58 0 2 

### 59 0 0 2 60 0 0 0



## -152­ 418Addendum 2·59 Twenty-eight·day repeated·dose oral toxicity study in rats BII·0908 Detailed clinical observations ofindividual animals (week I) Observations in arena Sex Exp. group AnimalNo. Tremor/twitch! Defecation Urination Stcreotypic Abnormal (mglkg/day) convulsion (count/min) (count/min) behavior behavior I 0 0 0 2 0 0 0 3 0 0 I 4 0 0 0, Vehicle 5 0 0 0 control 6 0 0 0 7 0 0 4 8 0 0 0 9 0 0 0 10 0 0 0 11 0 0 0 12 0 0 0 25 13 0 0 I 14 0 0 0
Male 15 0 1 16 0 0 0 17 0 0 12 150 18 0 0 2 19 0 0 0 20 0 0 0 21 0 0 0 22 0 0 0 23 0 0 2 24 0 0 I 1,000 25 26 0 0 0 0 0 0 27 0 0 2 28 0 0 0 29 0 0 0 30 0 0 0



## -153­ 419Addendum2-60 Twenty-tight-dayrepeated-doseoral toxicity study in rats BII-0908 Detailed clinical observations ofindividual animals (week 1) Observations in arena Exp. group AnimalNo. Tremorltwitcbl Defecation Urination Stereotypic Abnormal (mglkgldll)') convulsion (count/min) (countlmin) behavior behavior 

### 31 0 0 0 

### 32 0 0 0 

### 33 0 0 0 

### 34 0 0 1 Vehicle 

### 35 0 0 

### 0 control 

### 36 0 

### 0 0 

### 37 0 0 2 

### 38 0 0 

### 2 39 0 0 

### 0 40 0 0 

### 41 0 0 

### 0 42 0 0 0 

### 25 43 0 

### 0 2 

### 44 0 

### 0 7 

### Female 

### 45 0 0 0 

### 46 0 0 8 47 0 0 

### 0 ISO 

### 48 0 0 0 

### 49 0 0 

### 0 50 0 

### 0 0 51 0 

### 0 2 

### 52 0 0 0 53 0 0 

### 0 54 0 0 0 1,000 

### 55 56 0 0 0 0 

### 0 0 57 0 0 

### 0 58 0 

### 0 59 0 0 3 

### 60 0 0



## -154­ 420Addendum 2-61 Twenty-eight·dayrepeated-dose oral toxicity study in rats B11-0908 Detailedclinical observationsofindividual animals (week2) Observations in arena Sex Exp. group Animal No. Tremorltwitchl Defecation Urination Stereotypic Abnonnal (mglkglday) conVUlsion (count/min) (count/min) behavior behavior 0 0 1 2 0 0 0 3 0 0 4 4 0 0 0 Vehicle 5 0 0 0 control 6 0 0 0 

### 7 0 2 2 8 0 0 0 9 0 0 0 10 0 0 0 II 0 0 0 

### 12 0 0 0 25 

### 13 0 0 0 

### 14 0 0 0
Male 15 0 3 16 0 0 

### 2 

### ]7 0 0 5 1'50 18 

### 0 0 

### 0 19 0 

### 0 0 20 0 0 

### 21 0 0 

### 2 22 0 

### 0 0 

### 23 0 4 

### 24 0 1 

### 1.000 25 0 0 1 26 0 0 0 

### 27 0 2 1 28 0 0 0 

### 29 0 0 0 30 0 0 

### 0



## -155­ 421Addendum 2-62 Twenty-eight-dayrepeated-dose oral toxicity study in rats BlJ-0908 Detailed clinical observations ofindividual animals (week2) Observationsin arena Sex Exp. group AnimalNo. Tremorltwitch/ Defecation Urination Stereotypic Abnormal (mglkglday) convulsion (countlmin) (counllmin) behavior behavior 31 0 0 

### Q 32 0 0 0 

### 33 0 0 0 

### 34 0 0 0 Vehicle 

### 35 0 

### 0 0 control 

### 36 0 0 

### 0 37 0 0 0 

### 38 0 0 2 39 0 0 0 

### 40 0 0 I 41 0 0 0 42 0 0 

### 0 25 

### 43 0 0 0 44 0 0
Female 

### 45 0 0 46 0 0 2 

### 47 0 0 

### 0 150 

### 48 0 

### 0 0 

### 49 0 0 0 50 () 0 0 

### 5] 0 0 0 52 0 0 0 

### 53 0 0 0 

### 54 0 0 0 1,000 

### 55 56 0 0 0 0 0 

### 57 0 0 0 58 0 0 4 S9 0 0 

### 0 60 0 0 

### 0



## -156­ 422Addendum 2-63 Twenty-eight-dayrepeated-dose oral toxicity study inrats Bl1-0908 Detailed clinical observations ofindividual animals (week 3) Observations in arena Sex Exp. group AnimalNo. Tremorftwilcb/ Defecation Urination Stereotypic Abnonnal (mglkglday) convulsion (count/min) (count/min) behavior behavior I 

### 0 0 0 2 0 0 0 3 0 0 0 

### 4 0 0 I Vehicle 5 0 0 0 

### control 6 0 0 0 

### 7 0 3 I 8 0 0 0 9 0 0 0 10 0 0 

### 0 11 0 0 0 

### 12 0 0 0 25 13 0 0 0 

### 14 0 0 0
Male 15 

### 0 0 

### 0 16 0 2 

### 17 0 0 2 150 18 0 0 0 19 0 

### 0 0 

### 20 0 

### 0 0 

### 21 0 0 1 22 0 0 0 23 0 0 0 

### 24 0 ) 0 1,000 25 

### 26 0 0 0 0 0 0 

### 27 () 0 

### 4 28 0 

### 0 0 

### 29 0 0 0 30 0 0 0



## -157­ 423Addendum 2·64 Twenty-eight-day repcated-dose oral toxicity studyin rats BlJ-0908 Detailed clinical observations ofindividual animals (week3) Observationsin arena Exp. group AnimalNo. Tremor/twitch! Defecation Urination Stcreotypic Abnormal (mglkglday) convulsion (count/min) (count/min) behavior bellavior 31 0 0 0 32 0 0 0 33 0 0 0 34 0 0 0 Vehicle 35 0 0 0 control 36 0 0 0 37 0 0 3 38 0 0 0 39 0 0 0 40 0 0 2 41 0 0 0 42 0 0 0 25 43 0 0 0 44 0 0 0
Female 45 0 0 0 46 0 0 13 47 0 0 0 

### ISO 48 

### 0 0 0 

### 49 0 0 0 50 0 0 0 51 0 0 0 52 0 0 0 53 0 0 I 54 

### 0 0 0 1,000 

### 55 56 0 0 

### 0 0 0 0 57 0 0 0 S8 0 0 4 

### 59 0 0 0 60 0 0 0



## -158­ 424Addendum 2-65 Twenty-eight-day repeated-dose oral toxicity study in rats BI1-0908 Detailedclinical observationsofindividual animals (week4) Observations in arena Sex Exp. group AnimalNo. Tremorltwitchl Defecation Urination Stercotypic Abnormal (mglkglday) convulsion (count/min) (count/min) behavior behavior I 0 0 0 

### 2 0 0 0 3 

### 0 0 0 4 0 0 0 Vehicle 5 0 0 0 control 6 0 0 0 7 0 I I 8 0 0 0 9 0 0 0 10 0 

### 0 0

25 11 12 13 0 

### 0 0 0 0 0 0 0 0 14 0 0 0 Male 15 16 17 0 0 0 I 2 0 0 1 0 150 18 0 

### 0 0 19 

### 0 0 0 20 0 0 0 

### 21 0 ] 0 

### 22 0 0 0 23 0 0 

### 0 24 0 0 1,000 

### 25 

### 26 0 0 0 0 0 0 27 0 0 

### 6 28 0 0 0 

### 29 0 1 0 30 

### 0 0 0



## -159­ 425Addendum 2-66 Twenty-eight-dayrepeated-doseoral toxicity study in rats Bll-0908 Detailed clinical observations ofindividual animals (week4) Observations in arena Sex Exp. group AnimalNo. Tremorltwitchl Defecation Urination Stereotypic Abnormal (mglkglday) convulsion (count/min) (count/min) behavior behavior 31 0 0 0 32 0 0 0 33 0 0 0 34 0 0 0 Vehicle 35 0 0 0 control 36 0 0 0 37 0 0 0 38 0 0 0 39 0 0 0 40 0 0 0 41 0 0 0 

### 42 0 0 0 25 43 0 0 0 44 0 0 0
Female 45 0 0 0 46 0 0 6 47 0 0 0 150 48 0 0 0 49 0 0 0 50 0 0 0 51 0 0 0 52 0 0 0 53 0 0 0 54 0 0 0 1,000 55 56 0 0 0 0 0 0 57 0 0 0 58 0 I 3 59 

### 0 0 0 

### 60 0 0 0



## -160­ 426Addendum 2-67 Twenty-eight-dayrepeated-dose oral toxicity study in rats Bll-0908 Detailed clinical observations ofindividual animals (Recoveryweek I) Observations in arena Exp.group Animal No. Tremorltwitchl Defecation Urination Stereotypic Abnormal (mglkglday) convulsion (count/min) (count/min) behavior behavior 6 0 0 0
Vehicle control 7 8 9 0 0 Q 2 0 0 0 0
Male 10 26 0 0 0 0 2 0 27 0 1 1,000 28 0 0 0 29 0 0 0 30 0 0 0



## -161­ 427Addendum 2-68 Twenty-eight-day repeated-dosc oral toxicity study inrats BII-0908 Detailed clinical observations ofindividual animals (Recovery week I) Observations inarena Exp. group AnimalNo. Tremor/twitch! Defecation Urination Stereotypic Abnormal (mglkgday) convulsion (count/min) (count/min) behavior behavior 

### 36 0 0 0 37 0 0 

### 3 Vehicle control 38 0 0 0 39 0 0 Female 40 0 0 0 56 0 0 0 57 0 0 0 1,000 

### 58 0 0 0 59 0 0 0 60 0 0 0



## -162­ 428Addendum 2-69 Twenty-eight-day repeated-dose oral toxicity study in rats BII-0908 Detailed clinical observations ofindividual animals (Recovery week2) Observations in arena Sex Exp.group Animal No. Tremor/twitchl Defecation Urination Stereotypic Abnormal (mglkglday) convulsion (count/min) (count/min) behavior behavior 6 0 0 0
Vehicle control 7 8 9 0 0 0 2 0 0 0 0 2
Male 10 26 0 0 0 0 0 0 

### 27 0 0 I 1,000 

### 28 0 0 0 29 0 0 0 30 0 0 0



## -163­ 429Addendum 2·70 Twenty-eight-dayrepeated·dose oral toxicity study in rats B1I·0908 Detailed clinical observations ofindividual animals (Recovery week 2) Observations in arena 

### Sex Exp. group Animal No. Tremor/twitch! Defecation Urination Stereotypic Abnonnal (mglkglday) convulsion (count/min) (count/min) behavior behavior 36 0 0 0 37 0 0 0 Vehicle control 38 0 0 0 

### 39 0 0 0
Female 

### 40 0 0 0 56 0 0 0 57 0 0 0 1,000 58 0 0 0 

### 59 0 0 0 

### 60 0 0 0



## -164­ 430Addendum 3-1 Twenty-eight-dayrepeated·dose oral toxicity study in rats Bll·0908 Reflex ofindividual animals (week4) Sensorimotorfunction

Exp. group Animal No. Approachcontact! Pinna Painresponse Pupillary Airrighting (mglkglday) touch response response (tail pinch) reflex reflex. 0 0 0 + + 2 0 0 0 

### + + 3 0 0 0 

### + + 4 0 0 0 + + Vehicle control 5 0 0 0 + 

### + 6 0 0 0 

### + + 7 0 0 0 

### + + 8 0 0 0 

### + + 9 0 0 0 

### + + 10 0 0 0 + 

### + 11 0 0 0 

### + + 12 0 0 0 

### + + 25 13 0 0 0 + 

### + 14 0 0 0 

### + + Male IS 0 0 0 

### + + 16 0 0 0 + 

### + 17 0 0 0 

### + + 150 18 0 0 0 + + 19 0 0 0 

### + + 20 0 0 0 

### + + 21 0 0 1'1 + 

### + 

### 22 1'1 0 0 

### + + 

### 23 1'1 0 

### !'I + 

### + 24 0 0 0 + 

### + 1,000 25 

### 26 0 0 0 0 1'1 1'1 + + 

### + 

### + 27 0 1'1 1'1 

### + 

### + 28 0 0 

### 0 + + 29 0 0 0 

### + + 30 1'1 0 1'1 

### + +



## -165­ 431Addendum 3-2 Twenty-eight-day repeated-dose oral toxicity study in

### rats Bll-0908 Reflex ofindividual animals (week

### 4) Sensorimotorfunction Sex Exp. group AnimalNo. Approach contact! PiMa Pain 

### response Pupillary Airrighting (mglkglday) touch response response (tail pinch) reflex 

### reflex 31 0 0 0 

### + + 32 0 0 0 

### + + 

### 33 0 0 0 

### + + 

### 34 0 0 0 

### + + Vehicle control 

### 35 0 0 0 

### + + 36 0 0 0 

### + + 

### 37 0 0 0 

### + + 

### 38 0 0 0 

### + + 

### 39 0 0 0 

### + 

### + 40 0 0 0 

### + + 41 0 0 0 

### + + 

### 42 0 0 0 

### + + 25 43 0 0 0 

### + 

### + 44 0 0 0 

### + + Female 

### 45 0 0 0 

### + + 46 0 0 0 

### + + 

### 47 0 0 0 

### + + 

### ISO 48 0 0 0 

### + + 

### 49 0 0 0 

### + + 50 0 0 0 

### + + 51 0 0 0 

### + + 52 0 0 0 

### + + 

### 53 0 0 0 

### + + 54 0 0 0 

### + + 1.000 

### 55 0 0 0 

### + + 56 0 0 0 

### + + 57 0 0 0 

### + 

### + 58 0 0 0 

### + + 59 0 0 0 

### + + 6() 0 0 0 

### + 

### +



## -166­ 432Addendum 

### 4-1 Twenty·eight-day repeated-dose oral toxicity study in rats 

### 811·0908 Grip 

### strength ofindividual animals (week

### 4) Sex Exp.group Animal No. Forelimb Hindlimb (g) (mglkg/day) Trial 

### 1 481 Trial 

### 2 424 Mean 

### 453 Trial 1 448 Trial 2 414 Mean 

### 431 2 413 397 405 458 407 433 3 

### 404 471 438 433 472 453 4 

### 382 472 427 414 435 425 Vehicle control 

### 5 6 403 493 417 388 410 441 414 371 437 379 426 375 7 387 386 387 440 443 442 8 455 424 440 412 385 399 9 429 401 415 442 478 460 10 345 381 363 381 459 420 11 

### 430 377 404 471 

### 484 478 12 

### 375 384 380 465 431 448 25 \3 

### 449 428 439 483 426 455 14 465 375 420 463 448 456
Male 

### 15 457 423 440 455 488 472 16 379 450 415 325 366 346 17 368 449 409 464 429 447 150 18 456 446 451 455 479 467 19 340 424 382 433 418 426 20 467 495 481 439 479 459 21 353 434 394 345 363 354 22 450 397 424 378 423 401 23 459 498 479 420 417 419 24 434 455 445 

### 412 396 404 1,000 25 26 414 43] 400 484 407 458 384 383 412 416 398 400 27 462 458 460 408 382 395 28 326 385 356 433 344 389 29 408 440 424 476 454 465 30 398 412 405 400 454 427



## -167­ 433

### Addendum 4-2 Twenty-eigbt-dayrepeared-doseoraltoxicity study inrats BII·090& Grip strength ofindividual animals (week4) Sex 

### Exp.group Animal No. Forelimb (g) Hindlimb (g) (mglkglday) 31 Trial 1 425 Tria! 2 394 Mean 410 Trial 1 

### 363 Trial 2 394 Mean 379 32 369 327 348 375 345 360 

## n 323 437 380 401 341 371 34 340 352 346 407 386 397 Vehicle 

### control 35 36 367 373 369 397 368 385 403 350 388 

### 375 396 363 37 396 407 402 351 362 357 38 406 382 394 439 417 428 39 318 365 342 332 382 357 40 322 371 347 363 386 375 41 364 424 394 346 327 337 42 321 376 349 393 388 391 25 43 372 372 372 333 360 347 44 335 399 367 367 356 362 Female 45 377 326 352 356 411 384 46 373 336 355 343 371 357 47 374 376 375 357 331 344 150 

### 48 383 343 363 387 373 380 49 369 421 395 404 

### 358 381 50 

### 425 349 387 403 442 423 51 332 

### 327 330 411 374 393 52 361 386 374 318 387 353 53 340 339 340 421 402 412 54 400 359 380 402 366 384 1,000 

### 55 56 312 307 336 348 324 328 

### 324 411 362 363 343 387 57 376 398 387 381 424 403 58 

### 324 357 341 404 425 415 59 397 422 410 392 377 385 60 359 329 344 375 360 368



## -168­ 434

### Addendum5-1 Twenty-eight-dayrepeated-dose oral toxicity study in rats BII-0908 Motoractivity ofindividual animals (week4) Sex Exp. group 

### AnimalNo. 

### Interval (min) (mglkglday) 

### 0-10 10-20 20-30 30·40 40-50 50-60 Total 17 116 134 23 18 2 310 2 

### 166 .55 15 64 

### 51 77 428 3 206 172 138 22 4 

### 0 542 4 249 121 71 57 91 114 703 Vehicle 5' 

### 211 136 

### 128 110 70 41 696 control 

### 6 177 71 

### 4 74 3 70 399 

### 7 9 28 117 39 

### 2 46 241 8 302 140 

### 92 74 118 173 899 9 179 85 50 55 99 49 517 10 227 148 45 50 4 38 512 11 

### 139 48 9 68 18 7 289 12 283 136 107 15 72 

### 46 659 25 13 

### 193 SI 

### 73 29 8 

### 106 490 14 84 48 29 149 173 91 574 Male 15 239 116 80 42 55 48 580 16 9 

### 74 11 

### 21 11 

### 6 132 17 207 135 51 63 67 100 623 150 18 150 170 94 106. 122 52 694 19 126 106 

### ISS 35 

### 87 159 668 

### 20 140 30 30 4 46 39 289 21 173 89 35 70 3 

### 371 22 371 157 197 102 232 154 1213 23 166 54 65 88 59 124 556 24 278 214 160 113 80 166 1011 

### 1,000 25 

### 26 251 295 176 226 

### 166 183 154 82 57 112 40 79 844 977 27 197 78 88 36 116 

### 65 580 28 272 223 91 94 72 

### 14 766 29 265 157 127 139 

### 107 9 804 

### 30 325 115 5 

### 0 6 0 451



## -169­ 435

### Addendum 5-2 Twenty-eight-dayrepeated-doseoral toxicity study inrats B1I-0908 Motor activity ofindividual animals (week4) Sex 

### Exp.group AnimalNo. Interval (min) (mglkglday) ()"10 

### 10·20 20·30 30-40 4()"50 50-60 Total 31 205 108 55 53 8 

### 38 467 32 317 123 106 36 

### 0 15 597 33 188 119 53 60 11 

### 432 34 261 261 152 83 

### 35 0 792 Vehicle 35 254· 56 59 0 17 33 419 control 

### 36 215 133 129 20 138 152 787 37 180 97 102 25 39 74 517 38 137 101 46 88 7 4 383 39 307 129 203 131 

### 20 122 912 

### 40 300 200 76 47 97 145 865 41 234 149 104 13 72 

### 20 592 42 258 177 197 193 144 85 1054 25 43 247 152 84 114 46 3 646 44 250 134 110 

### 0 119 135 748
Female 

### 45 46 317 142 340 84 256 4 80 7 66 0 0 ]0 1059 247 47 215 160 119 70 57 9 630 150 48 198 89 7 18 75 2 

### 389 49 211 134 57 22 33 4 461 50 208 136 46 84 88 26 588 51 268 158 97 93 88 34 738 52 353 268 69 204 

### 221 48 1163 53 294 135 93 92 31 108 753 54 250 214 154 185 8 

### 0 811 1,000 55 56 

### 350 324 203 190 3] ]57 159 15 0 114 

### 0 155 743 955 57 256 111 

### 59 132 81 89 728 58 204 155 206 118 [16 117 916 59 327 211 223 227 157 174 1319 60 278 148 62 0 

### 15 163 666



## -170­ 436811-0908 Addendum 5-3 Twcnly-eight-dayrepeated-dose oral toxicity study in rats Motor activity ofindividual animals (Recoveryweek 2) Sex Exp.group AnimalNo. Interval (min) (mglkglday) 0-10 10-20 20-30 30-40 40·50 50·60 Total 36 157 127 91 7 116 72 570

Vehicle control 37 38 39 128 12 235 6 5 188 25 63 33 9 40 7 2 60 35 0 45 

### 210 53 631
Female 40 56 180 250 51 92 21 2 0 0 8 44 24 261 412 57 204 87 43 34 91 32 491 1,000 58 226 124 3 157 60 102 672 

### 59 282 268 237 8 58 0 853 60 185 143 112 132 91 131 794



## -171­ 4371111-0908 

### Addendllll 6-1 Twenty-eight-day repeated-dose oral tOJicity 

### study in rats Body 

### weights of individual animals(g) Sex Exp.group Administration 

### period (lig/kg/day) AninI No. -2 1 ' 3 8 12 17 21 26 28 

### (days)



### ...... -.J t-:> I lIale Vebicle 

### control

25

150

1,000 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 

### 30 140.7 150.4 155.3 147.5 149.1 152.5 149.8 151.2 145.4 143.3 149.3 154.5 141.7 146.3 151.7 155.2 149.8 151.4 146.5 140.7 145.8 149.3 152.5 150.8 142.1 152.6 141. 6 150.8 143.4 

### 148.8 155.2 163.2 169.2 168.3 169 1 167.8 160.5 173.9 163.3 161.4 169.4 171.2 157.7 184.3 169.0 171.2 164.4 169.1 161.9 155.2 160.7 167.5 172.6 167.5 159.2 171.5 158.3 167.9 164.1**16~L** 5 169.8 182.8 187.2 183.6 188.0 182.8 112.1 193.3 174.5 174.4 187.7 185.7 169.8 180

..2 181.9 185.4 178.3 188.3 170.1 167.3 165.8 169.0 183.5 180.3 164.2 180.4 157.2 168.3 176.3**114.~.** 204.4 232.3 232.9 261,6 231. 8 266.5 226.2 257.5 239.2 279.9 228.3 258.4 201.7 224.4 238 1 273.8 216.2 245.9 208.1 232.7 226.0 250.8 221.4 253.4 204.4 228.7 235.3 266.7 225.5 253.1 225.2 249.5 213.3 243.1 240.8 280.5 201.1 224.2 201.9 229,2 198.4 

### 215.2 201.6 330.7 216.6 

### 1148.9 214.5 245.8 201.7 221.6 209.8 238.9 186 3 208.5 208.3 235.0 227.0 262.3**Z:()L.J._2~.4J** 

### __ 266.7 302.6 305.8 301.2 336.2 301.5 264.0 315.5 287.9 268.5 283.5 297.7 .1 17.8 Z90.B 277.3 279.5 341.3 258.7 31:1.1 

### 236.8 268.6 283.3 277.4 252.4 280.6 241.9 268.7 305.0**~4_** 295.9 331.0 335.5 335.7 359.6 330.8 295.3 356.7 321.7 296.1 310.4 321.5 290.4 349.1 319.0 301.6 306.0 3U.4 270.6**~9i·6** 251.0 

### 292.8 305.6 300.9 272.8 306.9 261.8 288.9 336.5**~14,4.** 322,0 359.5 369 9 374 9 396.2 356.3 332.4 381.9 349.0 319.9 334.6 351. 6 312.6 390.0 340.3 327.8 336.1 429.5 Z89.S**331.~** 264.0 

### 315.5 

### 328.3 322.3 305.9 334.0 284.0 314.2 362.4 301.0 330.4 368.2 380.0 386.2 404.4 368.2 347 0 392.3 357 B 330.9 336.8 361.7 320.4 403 0 354.0 336.3 340.0 445.0 295.6 

### 337.8 269.2 318.4­ 

### 339.4­ 330.4 317.2 342.4 288.9 320.7 373.6 305.1



## t; 00

### AddendUll 6-2 TWenty-eight-dar repeated-dose oral toxicity study in rats Dll-0908 

### Body weights of individual ani.als(g) Sex Exp.group (JIg/kg/day) Ani.al No. -2 1 3 8 12 17 21 

### 26 28 31 129.4 144.5 156.7 182.3 196.8 216.5 228.7 238.4 244.9 32 113.9 125.4 129.9 147.6 159.5 174.3 185.9 199.6 203.4 33 118.1 128.5 137.3 160.1 175.0 194.1 208.2 216.1 217.2 34 133.0 144.8 155.2 177.2 184.7 201.5 216.9 224.1 236.6 

### Vehicle 35 121.7 133.3 140.5 157.4 170.7 189.3 200.7 210.6 217.1 

### control 36 124.3 133.0 142.7 163.2 170.3 190.4 198.6 211.1 209.1 37 117.0 122.7 130.2 143.6 154.5 172 1 182.2 193.5 198.6 38 131. 9 142.4 152.2 169.1 184 4 196.5 209.7 217.1 226.3 39 119.0 132.2 138.0 158.2 168.7 185.8 193.2 195.9 204.5 40 129.4 138.7 146.6 162.6 175.7 195.2 209.6 218.4 219.1 41 129.5 139.2 144.4 171.7 186.3 197.6 209.6 214 .4 223.8 42 119.0 131.7 142.6 156.4 162.4­ 176.5 189.0 200.5 206.2 25 43 117.6 120.2 127.9 139.5 152.8 166.3 174.7 189.2 193.8 44 127.3 135.1 144.2 166.1 180.3 202.1 213.4 223.3 235.7 feJIale 45 122.3 135.3 144.3 153.5 164.7 173.0 183.8 192.1 198.6 46 112.6 122.2 126.3 142.9 155.7 175.9 187.2 197.9 201. 3 47 124.4 132.5 141.5 164.8 178.4 195.8 206.2 217.2 218.6 150 48 128.6 143.7 153.6 191.6 193.0 202.1 212.5 226.0 238.4 49 119.4 132.0 137.6 160.9 177.2 200.6 209 3 218.9 228.3 50 129.8 140.9 147.4 189.3 182.2 205.0 216.0 227.5 226.1



### ....... 143:-6-

----­ -.:j 51 127.1 135.5 173.4 193.0 220.9 --Z35.4 241.3 244.4 (jj 52 121.0 130.2 135.1 160.5 185.5 202.1 216.3 225.0 226.9 I 53 132.9 149.8 151.5 173.3 193.0 220.6 235.4 248.4 255.0 54 117.9 125.3 132.3 150.1 161.2 179.7 191. 2 197.5 197.2 1,000 55 122.4 129.9 136.7 158.8 174.0 197.1 206.0 214.2 225.6 56 117.3 127.4 135.7 156.5 167.8 184.8 193 3 204.7 215.2 57 123.7 136.8 148.8 176.4 192.8 211.1 22B.9 235.4 245.6 58 129.6 140.7 154.9 169.5 186.8 210.7 218.5 228.0 232.3 59 127.1 137.B 147.3 171.2 190.5 206.5 219.5 235.2 230.9 60 

### U:l.2 132.0 138.!L l.61._L.

__ 174.7**:t9~.** 202.3 220.0 222.6

.j::" W <0

### AddendUlD 6-3 Twenty-eight-day repeated-dose oral toxicity study in rats Bll-0908 

### Body weights of individual aniaals(g) Sex 

### Exp.group !tecover,. period (q/kg/day) 

### Ani.al 110. 1 5 10 14 

### (days) 6 7 372 .3 351.8 

### 395.5 375.3 428.6 403.1 444.3 421.6 

### Vehicle control 8 9 398.3 361.3 420.4 380.6 441. 4 400.4 453.7 412 9 lIale 10 334.8 359.2 381.2 401. 6 26 340.3 366.1 399.4 424.0 27 

### 292.6 313.2 339.0 361.2 1,000 28 29 323.0 377.4 346.4 395.4 377.7 418.5 399.2 444.7 30 304.8 311.6 329.7 350.8



### female Vehicle control 36 37 38 39 40 207.5 202.3 226.3 207.5 223.5 218.3 212.5 232.6 214.9 235.1 225.9 215.8 234.9 215.8 243.6 228.7 222.2 243.2 224.8 249.2



## ...... 1.000 56 57 58 59 60 216.3 240.5 232.4 233.8 

### 220.4 211.3 247.9 240 4 237.5 

### 232.8 :laS.2 247.0 242.4 244.5 

### a.35.4­ 229.7 254.6 246.0 248.6 240.2 -.:J**~** I**~**

### ~Addendua 7-1 TweDty-eight-day repeated-dose oral toxicity study in rats 811-0908­ rood intakes of individual aniaa!s(g/rat/day) Sex Exp.group Administration period (llIg,/kg/day)

-_._._- Anillal No. 1 3 

### 8 15 2Z 28 (days) 

### 1 17.7 

### 17.4 18.1 17.9 

### 18.3 17.0 2 19.0 22.1 

### 23.1 21.2 21.3 19.2 3 20.1 21.1 22.0 21.1 20.4 19.2 4 20.0 19.6 21. 2 21.6 22.7 22.5 Vehicle 

### 5 22.1 21.1 24.9 25.3 23 9 21.7 control 6 20.0 19.5 21.4 20.5 19.7 17.8 7 19.1 17.9 18.2 18.0 20 0 20.9 8 21.9 21.9 23.5 23.0 23.3 21.1 9 19.8 18.1 20.1 21. 0 21. 5 20 2 10 11 17.5 21.1 16.9 20.7 18.3 20.2 17.9 18.2 17,

..

### L 17.1 

### lU 16.5 12 21.2 20.7 21.1 21. 4 20.9 19.4 25 13 18.2 17.0 19.4 18.4 18.6 17.3 14 20.9 20.0 24.3 25.1 23.8 23.3 

### Kale 15 20.7 18.9 20.6 20.2 19.3 17.4 16 20.3 20.9 21.7 19.8 18.5 17.6 17 20.5 19.7 20.7 21. 5 20.9 19.0 150 18 22.0 21. 2 26.0 28.0 28.2 28.6 19 21. 3 18.1 20.3 20.7 20.2 17.5



### ......**~** 20 21 18.6 19.2 17.5 13.8 18.8 18.0 20.5 16.8 21.0 17.1 21.0 16.5 01 I 22 23 21.3 20.9 13.6 16.9 17.6 20.2 20.4 22.S 21.4 22.0 19.7 22.6 24 22.1 18.4 22.0 22 9 24.6 27.8 1.000 25 19.6 14.4 18.7 19 2 20.7 24.S 26 20.8 16.6 18.3 22.6 23.2 22.6 27 19.4 13.4 15.2 17.8 17.9 18.2 28 20.9 13.7 18.0 22.0 

### 25.5 23.6 29 20.8 18.5 22.8 23.7 24.4 22.1 

### 30 11L8 16.9 

### 19.5 

### 24.6 35.0 21.6**~ ~** 811-0908 AddenduDl 7-2 Twenty-eight-day repeated-dose oral toxicity study in rats Food intakes of individual aniaa)s(g/rat/day) 

### Sex Exp.group Adllinistration period (lIIg!.kg/day) Aniad No. 1 3 8 15 

### 22 28 (days)



# .... -.:J 0:. ( feaale Vehicle control



### 25

150

1,000 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 

### tilL 17.1 13.8 15.S 17.2 16.1 16.0 13.9 16.7 16.4 17.5 16.1 15.4 12.1 16.1 16.9 14.6 16.1 17.2 16.3 16.6 14.9 15.8 19.3 13.6 15.5 15.6 17.7 17.2 15.1 

### U

..4 17.1 12.1 15.1 15.9 13.6 15.7 12.3 16.2 14.3 14.9 14.4 14.6 12.2 15.4 16.4 13.2 14.5 16.2 13.7 14.4 12.9 11.8 12.9 10.0 12.0 12.2 14.8 16.0 13.1 

### 11.2 16.3 12.6 15.7 15.5 12.9 14 4 12.0 14.5 14.6 14.0 16.4 12.7 11.2 15.5 14.4 13.3 15.9 18.8 15.3 16.1 15.4 14.6 12.1 11.4 14.3 14.1 17.1 14.8 14.0 

### 1::t.4 14.1 11.4 15.3 14.4 13.1 12.2 11.1 13.6 12.3 13.0 14.8 11.8 10.4 14.3 12.9 13.3 13.8 12.5 15.4 14.8 16.1 15.9 16.7 14.3 16.1 16.4 15.5 16.2 15.9 

### 14.5 13.7 11.4 14.2 13.0 13.8 13.9 11 4 12.4 12.2 13.7 13.1 11.0 11.4 14.7 11.6 15.1 14.3 12.9 14.8 15.3 16 7 15.0 18.4 19.3 17.5 17.7 16.6 16.2 16.2 14 7 12.2 11.1 12.4 14.2 13.5 11.9 11. 5 12.3 11.B 12.3 13.3 11.0 11.2 14.5 12.5 13.8 13.2 13.6 12.4 13.8 14.5 14.4 19.0 18.2 17.9 15.8 16.0 15.6 15.0 15.1



# :t N

### Addendll. 7-3 Tytnty-eight-day repeated-dose oral toxicity study in rats 811-0908 

### lood intakes 

### Qf indIvidual anilals(g/rat/day) Sex 

### bp.group Recovery period (q/kg/day) Animal No. 8 

### 14 (days) 

# " 6 21.0 22.6 24.6 7 23.3 26.3 26.3 

### Vehicle 8 23.6 24.0 24.2 

### control 9 24.1 24.5 25.6 

### Kale 10 20.4 22.8 24.0 26 25.7 26.3 28.8 27 22.1 23.6 25.9 1,000 28 22.9 25.0 25.7 

### 29 23.0 23.1 27.2 30 19.9 22.4 25.8

I-' -.;j -.;j 1 

### 'e.ale Vehicle control

1.000 36 37 38 39 40 58 57 58 59 *lQ 17.7 15.6 16.1 15.3 17.3 16.6 16.8 18.3 18.1 

## n.! 16.5 14.6 16.2 14.8 17.7 18.4 17.1 18.1 17.5 

### 18.5 16.7 16.5 16.0 15.9 18.7 16.8 17.2 18.0 

### ULI 18,2**~ ~**  

### wAddendull B-1 Twenty-eight-day repeated-dose oral toxicity study in fats Bll-0908 Hematological data of individual animals Exp.group RBC tlBC lib Ht tlCV MeR MCRe Platelet Reticulo P T APTT Sex (llg/tg/day) Animal No. (xl04 / ,uL) (xlOa/,uL) (g/dL) (%) (fL) (pg) (g/dL) (xl04 / ,uL) (%) (sec) (sec) 1 800 150 15.9 47.9 59.9 19.9 33.2 108.3 2.5 18.1 30.9 2 755 121 15.2 45.5 60.2 20.2 33.5 102.8 2.7 17.3 28.0 3 742 109 15.1 44.1 59.4 20.3 34.2 105.8 2.4 18.3 27.4 4 683 118 13.8 40.8 59.8 20.2 33.8 96.6 3.0 16.7 26.1 Vehicle 5 763**_!~,!** 

### __________ l,.§_.:...L

____4 5 _·j,

_________**?.~_~_~** 

### _______

### J_U

____

### .J!.9 . 7 2.3**_Ul_~_'!** 

### _________**~L·_?** 

### _______ ________**Jl_~_c.!3** control 

## recov;r-;-­ 6 802 128 15.1 44.5 55.4 18.8 33.9 114.3 2.2 24.6 35.3 7 753 92 14.7 43.3 57.5 19.6 34.0 120.0 2.6 24.4 35.3 8 746 127 15.2 44.4 59.5 20.4 34.2 103.2 2.1 18.2 36.9 9 861 125 15.8 46.3 53.7 18.3 34.1 109.2 2.1 18.0 37.9 10 791 104 15.3 44.8 56.6 19.4 34.2 102.8 2.8 15.9 32.1 11 751 103 15.0 44.0 58.5 20.0 34.1 95.4 2.0 14.7 25.9 12 749 136 14.9 43.7 58.4 19 9 34.l 99.4 2.8 15.4 31.3 25 13 724 126 14.4 43.5 60.1 19.9 33.1 99.8 2.4 17.5 32.3 14 742 145 14.3 42.8 57.7 19.3 33.4 103.2 2.5 15.9 27.5 Kale 15 742 91 15.0 44.9 60.5 20.3 33.5 102.9 2.2 16.6 26.8 16 711 154 14.8 44.4 62.5 20.9 33.4 99.9 2.7 16.4 25.7 17 781 78 14.9 45.2 57.9 19.1 32.9 108.1 2.1 14.8 27.1 150 18 730 150 14.7 43.7 59.9 20.1 33.6 107.5 2.8 18.9 28.5 I 19 781 111 15.3 46.0 58.9 19.6 33.3 94.5 1.6 24.3 34.1 f-' 20 741 77 15.0 45.2 61.1 20.2 33.1 87.0 2.5 18.2 28.0 ....;j 21 842 134 16.0 47.9 56.9 19.0 33.3 111.6 ·Z.4 17.0 36.2 I 22 791 142 14.4 43.1 54.5 18.2 33.5 117.7 2.2 16.2 30.5 23 738 83 14.9 44.8 60.6 20.2 33.3 92.5 2.6 18.3 30.3 24 817 142 16.1 48.7 59.6 19.7 33.1 93.6 3.7 18.2 30.3 00

1,000



### __**~?** 

### ______**I§.~**

### _______!.QQ

_________ 

### l,..?__'..Q

_______**H..:..~**

### ___**~?_:_1!.** 

### ________**!_~_L** 

### _____1L.!!.

_______1 0

### l_!_L

___**?..:....~**

### ___________**l,.~--'_!..**

### _______**~_~.:_?**

### _______ Recovery 26 796 141 15.0 44.8 56.3 18.9 33.5 115.2 2.6 28.2 39.1 27 742 126 14.5 43.6 58.7 19.5 33.3 117.2 2.9 14.5 32.7 28 744 146 14.4 41. 6 55.9 19.3 34.5 109.7 2.8 19.4 36.8 29 731 127 14.6 43.5 59.4 20.0 33.7 103.1

. 3.3 17.7 38.1 30 789 

### 116 15.3 45.6 57.8 19.5 33.7 99.1 2.0 17.4 31.7**~ ~** **~**811-0908 

### Addendum 8-2 Twenty-eigbt-day repeated-dose oral toxicity study in rats Heaatoiogical data 

### of individual aniaals Exp.group RBC WIIC Hb Ht KCV MeR IICHC 

### Platelet Reticulo l' T APTT 

### Sex (I,/ltg/day) 

### Anillal No. (xlO'/tlL) (xl0z/,tlL) (g/dL) (%) 

### (fL) (pg) (g/dL) (xl04

### /,tlL) (%) 

### (sec) (sec) 31 743 103 14.8 43.2 58.2 19.9 34.1 106.4 2.0 14.0 27.1 32 774 102 15.1 45.0 58.1 19.5 33.5 99.6 

### 1.7 14.3 21.4 33 

### 771 113 15.1 44.2 57.3 19.6 

### 34.2 100.8 

### 1.9 15.8 22.6 

### Vehicle 34 741 105 14.7 42.9 57.9 19.9 34.3 104.5 1.6 16.1 28.2



### ___**~1** 

### _____7E!.?.

___..
___ ___ 

### I.1..J!..

_____.1..!..·

..L

______

### J?.9_:...?.

____.
__!_!1.c.2

________**~l..2** 

### ________**!.?_e.~1** 

### _____J_:_l:!.

__________**l:.!L~** 

### _____**~_~_._~**

### _______ control Recovery 36 

### 803 81 15.8 46.1 57.4 19.7 34.3 114.3 2.5 14.5 27.7 37 787 98 

### 14.8 42.3 53.7 18.8 35.0 140.2 '1.8 14.8 20.7 38 754 84 14.6 41.7 

### 55.3 19.' 35.1 122.4 

### 1.S 14.4 25.7 39 791 90 14.6 41.7 52.7 

### 18.4 35.0 116.2 

### 1.9 14.0 22.8 40 750 108 14.9 41.6 55.5 

### 19.9 35.&**1~9.9 2.~.**  

### 14.6 24.2 41 774 87 15.4 4S.4 58.7 

### 19.9 33.991.7 1.6 13.6 

### 26.5 42 767 57 15.1 44.8 

### 58.5 19.7**33~8** 

### 107.1 1.5 13.7 24.1 25 43 760 112 14.9 44.1 58.1 19.7 33.8 112.2 1.7 11.2 

### 25.9 44 742 74 14.1 41.9 56.5 

### 19.0 33.7 115.0 

### 2.1 15.3 28.0 Feaale 

### 45 46 

### 47 770 788 760 101 80 92 15.3 15.5 14.9 44.9 45.1 44.3**§8.~** 

### 57.S 58.3 19.8 19.7 

### 19.7 34.0 

### .4.4 33.7 130.3 

### 110;4 111.9 1.6 

### 1.6 

### 1.9 Ui..3 14.8 14.2 26.0 

### 23.9 24.6 150 48 750 92 14.4 42.9 57.2 

### 19.1 33.6 94.' 2.1 15.1 

### 24.7
I-' -l**~** 

### 49 50 51 740 739 737 77 143 103 14.4 15.4 14.5 42.4 44.8 42.9 57.3 60.7 58.2 19.4 2Q.8 19.7 33.9 34.3 33.8 102.2 119.9 117.6 1.7 1

..6 1.9 15.7 15.0 15.0 22.6 19.8 22.7 I 

### 52 759 127 15.0 44.5 5&.7 19.8 33.8 110.9 2.0 14.5 24.8 53 716 134 14.2 41.7 58.2 19.8 .4.1 119.3 2.2 14.3 23.4 54 727 134 14.0 41.1 56.6 19.3 34.1 108.9 1.8 13.5 19.7 1,000 55 749 93 14.9 43.5 58.1 

## .iiieoverY'-----------------------------------­ 19.9



## - 94.2 99.5 2.1 16.7 27.7 

## -----. 56 807 71 14.9 43.1 53.4 18.4 34 5 130.5 1.8 14.7 26.0 57 760 89 14.9 43.0 56.5 19.6 34.7 121.8 1.8 13.9 21.2 58 805 85 15.0 42.6 52.8 18.6 35.2 119.7 1.9 14.7 24.4 59 783 57 14.6 41.9 53.5 18.7 34.9 113.1 1.9 14.4 22.1 60 766 78 14.9 42.7 55.7 19.5 34.9**11~.4** 

### J.9 15.1 25.8**~ ~**  ()15

10

15

20

25

30 AddendUJI 8-3 

### Twenty-eight-day repeated-dose oral toxicity study in rats B11-0908 

### Heaatological data of individual ani.als I!xp.grollP Differentiation of leukocyte (") Sex (ag/kg/day) Animal No. Meutro £osino Baso LYlIPh 110110 LIlC 1 12.7 0.6 0.2 83.4 1.9 1.3 2 11.1 0.4 0.2 84.7 2.5 1.1 3 20.6 0.5 0.2 73.3 4.1 1.4 Vehicle 

### cOlltrol 4

## ie"COVeri15 4 16.4 0.4 0.2 80.3 2.9 O.B Q..:_!L

____**O_~** ____**.1.1_~** 

### _____4.2

___

## -L..1
6 12.0 0.5 0.2 82.B 3.5 1.1 7 26.7 0.8 0.2 68.7 2.3 1.2 8 18.0 0.4 0.3 74.9 4.0 2.4 9 10.7 0.5 0.2 84.7 3.0 1.0 

### 10.3 0.7 0.1 85.8 2.0 1.1 11 20.1 0.7 0.2 75.4 3.1 0.4 13 15.2 0.3 0.1 81.4 2.1 0.9 25 13 15.0 0.5 0.1 79.7 2.B 1.9 14 14.4 0.4 0.2 79.6 3.5 1.9 

### Hale 23.5 0.5 0.1 73.6 1.0 1.2 16 17.9 0.3 0.2 76.7 2.7 2.2 17 25.1 1.3 0.0 69.5 2.7 1.3 150 18 15.6 0.5 0.2 79.7 3.3 0.6 I 19 21. 6 0.6 0.1 72.6 4.1 0.9 I-' 19.9 0.7 0.1 76.0 2.5 0.8 00 21 17.3 0.5 0.2 78.2 2.3 1.4 I 22 28.6 0.9 0.1 65 6 4.0 0.7 23 21.4 0.5 0.2 73.3 3.9 0.6 24 21. 2 0.6 0.2 70.9 5.5 1.6 0

1,000 24.2 0.6



### _______ q..:..t

__**-1Q_~L_.** 

### __ 4.2 

### ••

___...2...:..!.

______ llecovery 

### 26 14.4 0.4 0.1 2.1 27 11.4 0.7 

## o 1 2.5 28 10.2 0.8 0.1 1.5 29 15.5 0.5 0.1 7 11

..5 

### 1.1 0



# t C)811-0908 

### Addendull 8-4 Twenty-eight-day repeated-dose oral toxicity study in rats Hematological data of individual ani.als Exp.group Differentiation of leukocyte (") In 

### (llg/kg/day) Anilal 

### Ho. Neutro Eosino IbO 

### L)"Iph Hono LOC 31 15.0 1.2 0.1 80.6 2.6 0,6 32 14.3 0.7 0.1 82.4 1.5 1.1 33 10.5 1.1 0.1 84.2 3.2 1.0 34 8.5 1.0 0.2 88.0 1.2 1.1 

### Vehicle 35 12.3 0.6 0.1 85.3 0:8 0.9 

## RecoV;;-ry----------------------------------------------------------,---­ control 36 9.8 1.4 0.1 85.4 2.1 1.1 37 16.0 0.6 0.2 78.8 2.9 1.4 38 18.1 1.2 0.2 77.3 :.L 1 1.1 39 10.1 2.3 0.1 83.1 3.6 0.8 40 20.0 1.3 0.3 73.7 3.1 1.6 41 20.4 1.3 0.1 74.8 2.6 0.9 42 14.8 1.0 0.1 82.2 1.3 0.7 25 43 13.9 0.9 0.2 82.4 1.6 1.0 44 20.3 0.7 0.1 75.3 2.6 0.9 Pelale 45 13.6 1.8 0.1 81.0 2.0 1.5 46 16.9 0.7 0.1 79.2 1.9 1.2 47 15.2 0.8 0.1 81.2 1.7 0.9 150 48 17.2 0.4 0.1 80.2 1.3 0.9



# ..... 49 27.6 0.8 0.1 68.9 1.8 0.7 00 50 6.1 0.7 0.1 90.2 1.3 1.6



# ..... 51 18.8 l.Q 0.1 76.6 2.7 0.8 I 52 15.2 0.5 0.2 78.7 4.5 0.8 53 23.0 1.7 0.2 69.8 4.4 1.0 54 11.9 3.2 0.1 80.4 3.7 0.7 1.000 55 18.6 0.7 0.1 74.0 3.6 3.0 

## iiicov'{;j:i-------------------------------------------------------------------­ 56 11.8 1.5 0.1 82.6 2.7 1.3 57 18.3 1.0 0.1 75.3 4.0 1.3 58 9.0 1.8 0.1 83.9 3.6 1.6 59 9.6 1.0 0.1 84 4 3.4 1.4 

### 60 

### 9.6 1.0 

### 0.1 85.9 2

.. 

## a 1.3



# t -....I

### Addendum 9-1 Twenty-eight-day repeated-dose oral toxicity study in rats 8l1-0908 

### Blood cbs.ical data of 

### individual animals Ixp.group AST ALI ALP 

### ChE y -GTP 

### T-Cho TO Glucose T-Protein 

### AlbUllin A/G ratio**s~** 

### (lig/kg/day) AninI 

### No. (1l1/L) (lUlL) (lUlL) (lOlL) (IlllL) 

### (llg/dL) (lig/dL) (mg/dL) (g/dL) (g/dL) 1 73 Z1 709 47 0.6 49 10 155 5.7 2.7 0.90 3 63 

### 18 518 75 0.8 54 47 158 5.8 2.8 0.93 3 63 16 665 46 0.9 52 133 178**~.1** 2.9 0.91 4 55 15 366 71 0.9 48 76 176 6.0 2.7 0.82 Vehide 

### control 5 59 20 537 83 0,7 58 131 169 6.0 2.8 0.88 

## hcovery------------------------------------------------------------------------.-----------------------------------­ 6 7 57 86 19 26 273 387 55 44 0.8 0.5 46 55 87 74 161 145 

### 6.0 6.1 2,8 2.6 0.88 0.74 8 69 26 333 61 0.8 48 67 138 6.0 2.7 0.82 9 58 22 327 47 1.0 43 39 160 6.2 2.7 0.77 

### 10 58 

### lit**_..3~ .~_6**  0.9 6,5 63 1$7 5.6 2.7 0.93 11 58 15 677 67 0.7 58 69 193 5.7 2.7 0.90 12 88 18 495 37 0.8 50 71 151 6.0 2.7 0.82 85 13 63 22 576 55 1.0 50 36 143 6.3 2.8 0.80 14 73 26 568 43 0.7 49 84 143 5.6 2.8 1.00 Hale 15 

### .lt316 555 68 0.9 43 

### 50 169 5.4 a.6 0.93 16 72 27 550 43 0.9 46 69 

### 148 5T 2.8 1.00 17 54 19 477 49 0.6 53 69 160 5.6 2.8 1.00 150 18 57 22 350 85 0.6 58 179 154 5.7 2.8**0~97**

....... 00 t-..:l 19 

### 10 21 58 59 72 24 20 43 632 647 590 42 5Z 56 0.9 1.0 1.0 54 51 71 60 36 29 158 190 165 5.9 5.9 6 3 2.S 3.0 3.1 0.90 

### 1.03 0.97 I 22 77 49 509 67 0 8 48 29 125 5.9 2.9 0.97 23 62 37 452 79 0.8 57 19 142 5.8 3.0 1.07 24 66 39 382 61 0.8 52 107 145 5.8 3 1 1.15 1.000 25 80 51 640 42 0.8 51 34 123 5.8 3.1 1.15**Recover,.-----~-----------------------------------------------------------------.. -----------------------~**  26 63 21 338 52 0.8 52 53 142 6.0 2.8 0.88 27 71 24 311 44 0.6 49 54 109 5.6 2.6 0.87 28 61 22 255 64 0.6 62 44 128 5.8 2.6 0.81 29 66 23 321 45 0.5 49 81 142 5 8 2.5 0.76 30 85 34 332 42 0.8 46 47 136 5.7 2.7 0.90**~ ~**  00

### Addendum 9-3 Tventy-eight-day repeated-dose oral toxicity study in rats BJI-090B 

### Blood chemical data 

### of individual animals Exp.group AS! ALT ALP ehB l'-GTI' 

### T-Cho TG Glucose T-Protein AlbU1llin A/G ratio 

### Su (lIg/kg/day) Auiaal No. (IU/L) (IlJ/L) (Io/L) (lUlL) (Io/L) (llg/dL) (mg/dL)**(~/dL)** (g/dL) (g/dL) 31 66 16 273 275 0.9 60 40 119 6.0 3.1 1.07 32 76 23 779 212 0.9 63 24 143 5.9 2.9 0.97 33 75 21 351 300 0.9 73 22 119 5.8 3.0 1.07 Vehicle 

### control 34 61 29 374 304 0.6 54 31 133 6.1 3.1 ],03 35 68 17 197 183 1.0 63 19 128 6.3 3.1 0.97 

## fecovery------------------------------------------------------------.--------------------------------------------------------­ 36 52 15 211 

### 277 0.6 66 20 114 6.3 2.9 0.85 37 66 20 334 600 1.0 76 78 118 8.1 3.0 0.97 38 77 21 173 484 1.0 49 13 116 6.6 3.3 1.00 39 67 18 161 488 0.8 85 26 112 6.1 3.0 0.97 40 58 18 217 449 0.8 65 44 151 6.5 2.9 0.81 

### 41 59 17 259 166 0.7 45 16 106**~:6** 2.9 1.07 42 62 16 242 224 0.7 48 16 105 5.9 2.8 0.90 25 43 59 21 312 281 0.9 59 20 138 6.2 3.1 1.00 44 57 18 250 328 0.9 43 35 121 5.9 3.0 1.03 felale 45 73 22 297 339 1.0 52 21 116 5.8 3.1 1.15 

### 4'6' 50 26 451 162 0.7 75 28 141 

### 5.9 3.1 1.11 47 57 15 303 370 O.H 45 26 128 5.9 3.1 1.11 150 48 47 12 271 267 1.5 54 22 153 5.9 2.9 0.97

....... 00 

### c.o 49 50 51 53 73 67 16 21 39 456 273 473 266 228 143 0.8 1.1 1.2 83 61 

### 71 25 28 32 143 128 132 5.7 6.3**-~.9** 2.8 3.2 3.0 0.97 1.03 1.03 I 52 92 68 260 119 0.8 13 23 132 6.6 3.2 0.94 53 61 31 344 145 0.8 84 23 125 6.3 3.2 1.03 54 55 38 210 86 1.0 64 28 131 5.8 3.2 1.23 1,000 55 54 23 362 132 0.8 84 23 150 6.3 3.2 1.03 

## Rec-o;;ry--------------------------------------------------------------------------------------

...-----------------------------------------------­ sa 76 16 212 167 1.4 77 44 146 5.8 2.8 0.93 57 60 21 194 212 1.4 98 lOB 139 6.3 3.0 0.91 58 67 20 203 342 0.8 86 34 115 6.4 3 0 0.88 59 69 19 242 470 

### o.B 78 14 140 6.2 3.0 0.94 60 80 23 Z32 286 0.9 68 28 115 5.9 3.0 1.03

..1:10 ..1:10 to1111-0908 

### Addendua 9-3 Twenty-eight-day repeated-dose oral toxicity study in rats Blood chemical data of individual animals I!xp.group BUN 

### Creatinine T-Bil 

### Ca IP 

### Ha K 

### Cl 

## &ex hlg/kg/day) Anilal Ko. (ag/dL) (mg/dL) 

### (ag/dL) (ag/dL) (lig/dL) 

### (aEq/L) (lIt:q/L) (mEq/L) 1 7.5 0.17 0.06 9.9 7.4 143 4.3 105.8 2 7.6 0.19 0.06 10.1 9.0 143 4.1 104.6 3 8.4 O. 

### :n 0.04 10.3 7.8 143 4.2 107.2 4 11.2 0.27 0.04 10.3 7.6 144 4.0 105.7 VehIcle 5 8.1 0.19 0.10 10.3 8.1 145 3.8 106.3 

## Riicoveri----------------------------------------------------------------------------­ control 6 12.2 0.21 0.06 10.0 7.1 142 3.7 103.8 7 13.5 0.23 0.07 10.2 7.4 143 3.6 104.0 8 14.3 0.22 0.06 9.7 6.6 142 4.0 104.5 9 13.7 0.22 0.06 9.9 7.3 144 4.0 106.5 10 16.9 0.23 0.07 9.7 6_.8 141 4.1 103.7 11 10.7 0.22 0.08 9.9 7.6 143 4.3 104.8 12 7.9 0.24 0.04 9.9 7.3 144 4.4 107.8 25 13 8.9 O. Z1 0.07 10.2 7.1 145 4.1 107.6 14 6.7 0.15 0.05 10.2 7.7 144 4.2 106.2 Kale 15 10.7 0.24 0.06 10.3 9.2 145 4.8 107.8 16 8.0 0.17 0.09 9.8 6.8 142 4.3 105. 

### i-­ 17 9.6 0.22 0.06 9.8 7.3 143 4.2 106.7 150 18 9.5 0.21 0.05 10.4 9.1 143 4.3 104.5 I 19 6.8 0.21 0.06 10.3 8.4 144 3.9 106.0



### ...... 20 11.0 0.20 0.07 10.3 9.2 144 4.2 105.0 00 !I>- 21 17.7 0.29 0.14 10.1 9.4 141 4.3 105.2 I 22 19.2 0.31 0.13 9.9 9.3 142 4.3 106.1 23 9.0 0.20 0.07 9.9 9.2 144 4.5 107.9 24 12 .0 0.24 0.08 9.9 9.4 144 4.3 107.4 1,000 25 10.6**0.~4** 0.08 10.3 8.6 144 3.9 108.8 

## Recoveri---------------------------------------------------------------------------------------­ 26 13.9 0.19 0.05 9.7 7.3 144 4.3 105.9 27 17.5 0.21 0.09 9.7 6.8 140 4.4 104.2 28 12.6 0.19 0.08 9.6 6.8 143 4.3 105.7 29 14.3 0.20 0.08 9.7 8.2 143 4.3 103.2 3.0 

### 13.1 

### ILJ.J3 0.06 

### S.l 6 •.6 144 3.9 

### 105.3**~** OJ 

### o

### Addendull. 9-4 Twenty-tight-day repeated-dose oral toxicity study in rats B11-0908 

### Blood cheaical data of individual anilals l!xp.group BUN Creatinine T-Bil 

### Ca JP lIa K Cl S.I 

### (lIIg/kg/day) Anilal No. (mg/dL) (mg/dL) (llg/dL) (ag/dL) (lIg/dL) 

### (IIEq/L) (IliEq/L) 

### (dq/L) 31 10.2 0.22 0,07 9.9 6.7 144 3.6 10B.6 32 9.1 0.22 0.08 9.9 B.9 142 3.9 107.3 33 11.8 0.24 0.04 9.7 6.7 143 3.9 107.3 34 10.6 0.23 0.04 9.8 6.6 144 4.0 111.4 

### Vehicle 35 9.2 0.23 0.07 10.1 7.8 142



### _____ ________l.Q]...:..!L

____ control Recovery 36 19.0 0.27 0.07 9.8 4.8 141 3.8 108.0 37 13.2 0.26 0.09 9.9 5.0 141 3.4 103.9 38 21. 2 0.37 0.06 9.8 5.2 142 37 106.4 39 16.3 0.26 0.13 9.6 5.5 142 3.8 107.0 40 15.4 0.26 0.09 9.9 5.5 142 3.7 106.7 41 8.7 0.19 0.03 9.5 6.4 143 3.6 108.6 42 11.0 0.28 0.05 9.2 6.6 142 4.1 110.0 25 43 13.5 0.25 0.04 10.0 7.5 143 3.9 109.4 44 9.4 0.20 0.04 9.9 6.4 144 3.6 108.6 

### lemale 45 14.4 0.27 0.05 9.9 7.3 144 3.,7 110.3 46 14.2 0.19 0.09 10.8 9.1 142 4.4 ---'103.7 47 9.3 0.18 0.05 10.1 7.3 142 4.0 106.7 150 48 9.8 0.21 0.06 9.8 7.9 143 3.8 108.3 49 7.3 0.21 0.04 9.7 6.4 142 4.3 10B.9 I-' 50 10.5 0.25 0.09 9.9 6.3 143 3.8 108.4 00 01 51 8.0 0.28 0.08 10.1 8.9 143 4.3 106.5 I 52 11.3 0.26 0.08 10.3 8.8 145 3.7 107.0 53 8.5 0.22 0.08 10.1 9.0 144 3.9 107.3 54 15.9 0.30 0.10 9.5 8.3 131 4.3 98.6 1,000 55 11.6 0.26 0.10 10.3 B.8 140 3.8 103.1 

## Il.ecOvery----------------·---·------------------------------------------------------.------------------­ 56 14.6 0.24 0.07 9.5 6.4 143 4.3 109.7 57 21.6 0.26 0.13 10.2 6.8 140 4.7 104.6 58 14.4 0.22 0.08 10.0 5.9 141 3.9 104.6 59 13.6 0.24 0.07 9.5 5.7 144 3.3 107.0 

### 60 

### 18.8 0.30 

### 0.08 9.5 5.9 142 4.0**IJ~.9.** 

### 6**~** 01

### Addendum 10-1 

### TWentY-eight-day repeated-dose oral toxicity study in rats Bll-0908 

### Urinalytic data of indiTidual aniaa18 Exp.group 

### Urine volUie Uosa Sex 

### (a,!kg/day) Ani_I 110. (1111) 

### (lIllsa/L) 1 9 536 2 20 250 3 10 613 4 4 1914 

### Vehicle 5 23 276 

### control Recovery 6 16 569 

### 7 11 637 8 6 1193 9 9 852 10 5**17~2** 11 3 2040 12 5 

### 1211 25 13 3 2138 14 7 876 !lale 15 

### 3 2276 

### ---------_
....

# ­

16 2 2032 17 2 3034 150 18 8 1411 19 9 532



### ...... 20 6 1048 00 21 6 

### 970 Q':J 22 8 615 I 23 12 726 24 20 308 1.000 25 29 217 

## ----------------_......

# _ .....

## ­ Recovery 26 15 633 27 16 656 28 13 653 29 6 1437 30 18 378**~** CJ1 I'.)

### AddendWi 10-2 

### Twenty-eicht-day repeated-dose oral toxicity study in rats Bl1-0908 

### Urinalytic data of individual animals 

### Rxp.group Urine volua8 Uosm Sex (Ig/kg/da,) Anilal 110. (IlL) 

### (IOsa/L)

Vehide 

### control 31 32 33 34 35 

### Recovery 36 37 38 39 40 11 2 2 7 5 8 7 5 2 6 563 2204 2566

694 1017

779 928 1069 2472 la63



### ....... Pellie 25

150 41 43 43 44 4§ 46 47 48 49 50 18 2 1 10 

### zg

'1 4 3 8 5 263 1796 1461

474 

## U3 692 1244 1225

626 1037 00 51 38 85 -l 52 32 168 I 53 14 545 54 13 555 1,000 55 9 886 

# -------_

...

# _-­ Recover, 56 7 862 57 4 1556 58 5 1356 59 2 2302 6Q 3 1490**~** CJ'1 W611-0908 Addendum 10-3 Twenty-eight-dayrepeated-dose oral toxicity study in rab Urinalytic data ofindividual animals Exp.group Color Turbidity pH Protein Glucose Occult blood Sex (mg/kg/day) Animal No. 

### 1 Y NT 6.5 

### 1+ 2 SY NT 7.0 

### ± 3 

### Sy NT 6.5 1+ 4 Y NT 6.0 

## z+ Vehicle 5 SY NT 7.0 

### ± control RecoverY 6 SY NT 7.0 1+ 7 SY NT 7.0 1

### + ± 8 Y NT 6.5 1+ 9 SY NT 6.5 

### ± 10 Y NT 6.5 1+

# n12 

# y Y 

### JIlT NT '6.0 6.5 

### 2+ 1+ 25 13 Y NT 6.0 2+ 14 Y NT 6.5 1+ Male 15 16 Y Y NT NT 6.0 6.0 

## z+ 2+ 17 Y NT 5.0 2+ 150 18 Y NT 6.0 2 19 SY NT 6.5 1 I-' 20 Y NT 6.5 1+ 00 00 I 1,000

SY, Slightly yellow. Y. Yellow. NT. No turbidity. 21 22 23 24 25 Recovery 26 27 28 29 30 Y SY SY SY SY SY SY SY Y SY NT NT NT NT NT
NT NT 11<1 NT NT 6.0 1+ 6.0 1+ 6.0 1+ 6.5 

### ± 6.;:,.5

____

## = 7.0 7.0 7.0 6.5 7.0 ± 

### ± 1+ 1+ 

### ±**~** (J'I**~**611-0908 Addendum 1(}-4 Twenty-eight-day repeated-dose oral toxicity study in rats Urinalytic data ofindividual animals

Sex Exp.group (mg/kg!day) Animal No. Color Turbidity pH Protein Glucose Occult blood

Female Vlit,lic\tt 

### control

25 

# u 32 33 34 

### 35 &iCOV81j'. 36 37 38 39 40 

### 41 42 43 44 45 

# w Y Y SY Y
SY Y Y Y Y $V Y Y SY SY**~** NI' NT NT NT
NT NT NT NT NT 

### NT NT NT NT NT 

## u ± 5.0 2+ 5.0 2+ 6.5 1+ 

### 6.5

__..!.l+.!-

_____________ ,._­ 6.5 

### ± 6.5 1+ 6.5 1+ 6.0 2+ 6.5 1+ 

### 7.0 .::t 6.0 2+ 6.0 Z+ 6.5 1

## + 7.0

........ 00 <.0 I 

### 150

1.000 46 47 48 49 50 

## n**~ ~** **~** H SY Y Y Y Y 

### SY SY SY 

### SY SY NT NT NT NT NT 

### NT NT NT NT NT 6.5 6.0 6.5 6.5 6.0**7~** 

### U U 

### U U 1+ 2+ 1+ 1+ 1+


### ±

1+ Recovery 56 57 

### 58 59 60 SY Y Y Y 'Y NT NT NT NT NT 7.0 6.5 6.5 6.5 6.5 

### ± 1+ 1+ 2+ 1+ SY. SlightlyyeUow. Y, YliUow. NT. NQ turbidity.

oj:>. CJ'l CJ'l811-0908 Addendum 10-5 Twenty-eight-day repeated-dose oral toxicity study in rats Urinaiytic data. ofindividual anima.Js (Urinary sediment)



### ...... (0 

## o I Exp.group (mg/kg/day) 

### Sex

Mala Animal No. -I-­ 

### Z 3 4 Vehicle 5 control Recovery 6 7 8 9 

## to If 12 25 13 14 15 

### 16 17 150 18

1.000 19 20 21 22 23 24 Red blood ceUsal


## a 0 0 0 0 0 0 0 0 Q 

### 0 0 0 0 0 

### U 0 0 0 0 0 0 0 0 White blood cells') Epithelial cells,l

0 0 0 2 0 3 0 0 0 

## -----------_. 0 0 0 2 1 0 0 2 0 0 

### 00--­ 0 1 0 0 0 1 0 0 

### 0 I 0 0 0 0 0 3 0 0 14 18 5 33 11 84 6 11 CMtS')

0 0 0 0 0 0 0 0 0 0 

### 0 0 0 0 0 

### 0 0 0 0 0 0 0 0 0 Crystals')



### + 

## +
± 

## + 

## + ± j::

27 28 

### 29 30 

## o o o 

## o o o o o o 

## o 2 

## o 2 1 

## o o 

## o 

## o o 

## o ±
a) Number ofcells/l0 views (x400). b) Number ofcasts/18X 18 mm2• c) Incidence ofcrystals/I8 X 18 mmz

.

.". 

### c.n C)Addendum 10-6 Twenty-eight-dayrepeated-dose oral toxicity study in rats Bll-090S Urinalytic data ofindividual animals (Urinary sediment) Exp.group Red blood cells·) White blood cells') Epithelial cells.,) Castsbi CrystalsC ) Sex (mg/kg/day) Animal No. -31-­ 32 33 34 Vehicle 35 control 37 38 39 40 4I 42 25 43 44 Female 45 45 47 

### ISO 48



## ...... 49**~** 50



# ..... I -61 52 53 54 1,000 Recovery 56 57 58 59 60 a) Number ofcells/lOviews (X400>' b) Number ofcuts/ISX 18 mm2• cllncidence ofcrystals/ISX 18 mm 2•**~** U1



## ..... 0 0 0 0 0 0 0 0 0 0


### o 0 0 2 0 I 0 0 0 I 0 0­ () 1 0 0 0 1 () () 0 0 0 

### 0 0 1 0 0 

### 0 0 

### ·00 0 0 

### 0 0 0

0 0 0 2 0 0 0 0 0 0 2 1 3 1 3 0 1 1 Z 1**O~~** 0 2 0 1 0 1 0 0 ()**~** 0 0 () ! () ++ 0 () 4 () 9 0 4 12

1 4 3 2 2 

## o o o 

## o

0 0 0 0 :t 0

0 0 () ()

0 0 

### + 0 0 :t 0 +Addendum 11-1 Twenty-eigbt-day repeated-dose oral toxicity study in rats Bll-0908 Absolute organ weights of individual 

### anilals

Sex Exp.group (llg/kg/day) Animal No. Liver (g) Heart (g) Iidncy (g) Testis (g) Epididymis (g) Ovary (lIIg) Brain (g) Spleen (g) Thymus (lIg) Adrenal (mg) Body lIeight (g)

. .. . -----.-...--. 1.85 0.63 466.2 41.3 313.2 2 11.21 1.15 2.32 2.63 0.76 1.82 0.60 802.4 44.2 352.1 3 11.60 1.20 2.48 3.19 0.77 2.08 0.55 540.4 53.3 356.9 4 12.75 1.15 2.31 3.42 0.76 2.04 0.81 640.0 58.0 363.2**hhlc~** control 5 12.28 1.19 2.54 3.24 0.74 2.07 0.67 594.9 53.7 383.4 

## RecoVilj.·i--·---------------------------------·-·

---..------.--------------------.----.---------------------------------------.-------------------­ 6 11.48 1 33 2.69 3.03 1.02 2.14 0.67 663.2 50.7 423.8 7 10.64 1.25 2.73 3.36 1.18 2.27 0.63 513.3 48.3 393.4 8 11.44 1.41 3.02 3.08 1.12 2.10 0.79 557.1 58.0 427.5 9 11.23 1.44 2.89 3.61 1.17 2.13 0.70 414.7 56.1 380.1 10 10.00 1.26 2.75 3.25 1.01 2.02 0.59 499.5 54.9 374.2 11 10.96 1.03 2.343.37 0.82 1.98 0.54 6306' 52.1 323.7 12 11.99 1.25 2.56 3.21 0.83 2.00 0.70 533.2 47.3 344.0 25 13 9.54 1.09 2.17 2.93 0.71 1.95 0.66 508.7 50.0 302 5 14 10.76 1.15 3.00 3.02 0.84 1.95 0.64 733.0 44.2 383.8 Kale 

### 15 lO.19 L15 Z,S7 

### .a.21 Q.S.1. 1.94 0.58 

### 621

..._; 45.8 337.1 16 10.0Z-----1.15 2.72 2.92 0.66 1.90 0.67 570.5 38.0 318.4 17 10.98 1.01 2.24 2.95 0.73 2.00 0 43 282.4 39 

### 6 324.1 150 18 13.84 1.38 2.58 3.04 0.73 2.03 0.67 668.2 52.7 420 7**~** 19 9.24 1.02 1.95 3.03 0.78 1.96 0.48 380.9 42.6 280.4 >.0 20 10.52 1.23 2.30 3.06 0.85 

### Z.09 0.60 636.5 39.4 322.7 tv I 21 22 10.02 10.82 0.98 1.14 2.28 2.85 2.93 3.13 

### 0.66 0.75 1.80 1.BO 0.38 0.44 300.6 362.5 33.4 44.5**24~** 290.7 23 11.73 1.15 2.95 3.33 0.87 1.82 0.S2 369.4 47.1 310.3 24 12.57 1.13 2.69 2.77 0.65 1.92 0.66 420.4 41.7 301.9 1.000 25 10.58 0.97 2.79 3.38 0.78 1.88 0.39 413.2 42.8 282.3 

## RecOvery----------------------------------------------------.-------------------------

..-------------------------.--------------------------------­ 26 11.74 1.37 3.14 3.06 0.86 2.04 0.70 442.6 46.1 386.3 27 9.33 1.13 2.91 3.08 0.91 2.01 0.54 372.7 40.4 333.7 28 10.63 1.17 2.86 3.42 0.94 1.96 0.56 601.0 54.9 371.5 29 12.51 1.50 3.31 3.04 1.01 1.97 0.77 539.2 57.2 414.9 30 8.48. 1..:12 2.69 2.92 0.91 1.99Q.SL 305.9 37.1 3aa.6

.j». CJ'1 CO811-0908 Addendum 11-& Twentr-eight-day repeated-dose oral toxicity study in rats 

### Absolute organ weights of individual animals


### Sex Exp.group Liver Heart Kidney Testis EpididYliis Ovary 

### Brain Spleen Thymus 

### Adrenal Body weight (mg/kg/day) Animal No. (g) (g) (g) (g) (g) (Ilg) (g) (g) (lIg) (mg) (g) 31 7.20 0.96 1.69 57.1 1. 86 0.37 535.1 54.8 234.1 32 5.61 0.68 1. 58 76.4 1. 82 o 39 425.6 61. 9 190.1 33 6.52 0.83 1.48 81.1 1.89 0.53 .537.2 58.8 201.7 34 6.22 0.80 1 64 83.4 1. 90 0.41 555.5 64.0 220.6 Vehicle 35 6.50 0.79 1.43 82.5 1.62 0.40 547.5 54.3 208.8 

# iii-covery

----------------------------------------------------------------------------------------------------------------------------------­ control 36 6.02 0.82 1.43 74.1 1.83 0.49 488.4 57.2 217.3 37 5.67 0.74 1.48 66.7 1.93 0.45 403.3 57.7 209.9 38 6.02 0.77 1. 56 88.7 1.84 0.44 330.4 66.5 232.6 39 5.71 o 84 1. 65 82.3 1.96 0.48 408.0 65.6 212.9 40 6.20 0.83 1.61 72.6 1.85 0.45 436.2 52.0 230.9 41 42 6.42 5.17 0.86 0.65 i-.69 1.39 83.7 66.3 1.95 1. 86 0.46 0.33 655.8 453.3 65.8 64.0 214.1 197.0 135 43 5.85 0.75 1. 35 69.6 1.75 0.32 504.8 59.0 183.2 44 6.79 0.72 1. 65 60.1 1 .97 0.35 431.9 72.9 224.7 Pellale 45 5.40 0.65 1.17 74.6 1.71 0.38 379.8 53.8 189.0



## ..... <:0 150 46 47 48 49 50 6.07 8.02 7.43 7.33 

### 6.92 0.72 0.90 0.83 

### 0.86 0:.81 1. 53 1. 74 1.68 l..67 1.45 65.2 78.7 76.6 80.5 83.4 

## -**1~78** 1.86 1.88 1.93 1. 87 0.37 0.51 0.39 0.48 0.41 414.3 345.9 433.0 408.9 556.1 50.7 88.5 64.7 64.7 74.6 190.2 ZOB.5 219.4 214.3 218.1 CO I 51 52 9.67 9.11 0.89 0.83 2.13 1.81 83.2 65.4 1.82 1. 82 0.48 0.47 410.3 373.9 48.8 70.2 230.7 210.4 53 10.63 0.93 2.49 71.7 1.88 0.38 371.7 72.7 230.3 54 7.57 0.75 1.77 98.8 1.82 0.41 366.9 55.3 189.6 1,000 55 9.52 0.85 2.04

- 78.1 1.86 0.46 343.0 57.1 203.9 

## ie.;-ov8r;-------------------·------------------------------------------------------------------------------------------------------------------------­ 56 6.18 0.81 1.63 83.4 1. 87 0.44 398 4 61.7 210.6 57 7.47 1.05 1. 68 73.3 1. 76 0.55 443.6 61.0 241.0 58 6.88 1.03 2.04 84.4 1.85 0.51 482.3 60.1 233.1 59 6.84 0.90 1.88 94.5 1.91 0.60 455.1 73.5 234.0 60 

### 6

..14 .0.88 

### 1.56

-----­ 

### 81.2 l.U 0.5a 345.0 75.5 224.4

.j:I. CJ1 co811-0908 Addendum 12-1 Twenty-eight-day repeated-dose oral toxicity study In rats 

### Relative organ 

### weights of iodividual animals

Sex Exp.group (Ig/kg/day) Animal No. Liver (g/IOOg) Heart (g/100r;) Kidner (g/lOOg) Testis (g/IOOg) Epididyllis (g/IOOg) Ovary (lg/l00g) Brain (g/100g) Spleen (g/100g) ThYIUS (lg/100g) Adrenal (mg/IOOg) Body weight (g)



### Vellicle cootrol 1 2 3 

### 4 5 Rec'overy 6 7 

### 8 9 10 3.07 3 18 3.25 3.51 3.20 2.71 2.70 2.68 2.95 2.67 0.38 0.33 0.34 0.32 0.31 0.31 0.32 0.33 0.38 0.34 0.75 0.83 0.22 0.59 0.20 148.9 13-:2 313.2 0.66 0.75 0.22 0.53 0 17 227.9 12.6 352.1 0.69 0.89 0.22 0.58 0.15 151.4 14.9 356.9 0.64 0.94 0.21 0.56 0.22 176.2 16.0 363.2 0.66 0.85 0.19 0.54 0.17 155.2 14.0 383.4

--------------------------------------------------------------------------------------------------------­ 0.63 0.71 0.24 0.50 0.16 156.5 12.0 423.8 0 69 0.85 0.30 0.58 0.16 130.5 12.3 393.4 0.71 0.72 0.26 0.49 0 18 130.3 13.6 427.5 0.76 0.95 0.31 0.56 0.18 109.1 14.8 380.1 0.73 0.87 0.27 0.54 0.16 133.5 14.7 374 2



### ......**~** Kale 25

150 11 12 13 14 15 16 17 18 19 20 3.39 3.49 3.15 2.80 3.02 3.15 3.39 3.29 3.30 3.26 0.32 0.36 0.36 0.30 0.34 0.36 0.31 0.33 0.36 0.38 0.72 0.74 0.72 0.78 0.70 0.85 0.69 0.61 0.70 0.71 1.04 0.93 0.97 0.79 0.95 0.92 0.91 0.72 1.08 0.95 0.25 0.24 0.23 0.22 0.24 0.21 0.23 0.17 0.28 0.26 0.61 0.58 0.64 0.51 0.58 0.60 0.62 0.48 0.70 0.65 0.17 0.20 0.22 0.17 0.17 0.21 0.13 0.16 0.17 0.19 194.8

. 155.0

168.2 191.0 184.4 179.2

87.1 158.8 135 

### B 197.2 16.1 13.7 16.5 11.5 13.6 11.9 12.2 12.5 15.2 12.2 323 7 344.0 302.5 383.8 337.1 318.4 324.1 420.7 280.4 322.7 ,p. I
1,000 21 4.08 0.40 0.93 1.19 0.27 0.73 0.15 122.4 13.6 245.5 22 3.72 0.39 0.98 1.08 0.26 0.62 0.15 121.7 15.3 290.7 23 3.78 0.37 0.95 1.07 0.28 0.59 0.17 119.0 15.2 310.3 24 4.16 0.37 0.89 0.92 0.22 0.64 0.22 139.3 13.8 301.9 25 3.75 0.34 0.99 1.20 0,28 0.67 0.14 146.4 15.2 282.3 

## iie-cover-i----------------------------------------------

..-----------------------------------------------------------------------------.-------­ 26 27 28 29 30 3.04 2.80 2.86 3.02 2.58 0.35 0.34 0.31 0.36 0.37 0.81 0.87 0.77 0.80 0.82 0.79 0.92 0,92 0.73 0.89 0.22 0.27 0.25 0.24 0.2.S 0.53 0.60 0.53 0.47 0.61 0.18 0 16 0.15 0.19 0.16 114.6 111.7 161.8 130.0

93.1 11,9 386 3 12.1 333.7 14.8 371,5 13.8 414.9 11.3328.6



### ""­ 0) 

### o811-0908 

### Addendull 12-2 Twenty-ei,ht-day repeated-dose oral toxicity study in rats Relative organ weights of individual ani.als

Sex 

### Exp.group Liver Heart Kidner Testis 

### Epidid)ll!is Ovary Brain Spleen ThYlius Adrenal Body weight (q/kg/day) MiDI 110. (,/IOOg) (g/IOO,) (g/IOOg) (g/IOOg) (g/100g) (llg/IOO,) (g/100g) (g/IOOg) (llg/100g) (llg/100g) (g) 31 3.08 0.41 0.72 24 4 0.79 0.16 228.6 23 4 234.1



### Vehicle control 32 2.95 0.36 0.83 40.2 0.96 0.21 223.9 32.6 190.1 33 3.14 0.40 0.71 39.0 0.91 0.26 258.6 28.3 207.7 34 2.82 0.36 0.74 37.8 0.86 0.19 251.8 29.0 220.6 35 3.11 0.38 0.68 39.5 0.78 0.19 262.2 26.0 208.8 

## iiicovii-ry------------------------------------------------------------------------------

..-.------- ...-------------------------------------­ 36 37 2.77 2.70 0.38 0.35 0.66 0.71 34.1 31.8 

## o 84 0.92 0.23 0.21 224.8 192.1 26.3 27.5 217.3 209.9 38 2.59 0.33 0.67 38.1 0.79 0.19 142.0 28 6 232.6



## ..... <0 Fellale 25

150 39 40 41 42 43 44 45 46 47 48 49 50 2.68 2.69 3.00 2.62 3.19 3.02 2.86 3.19 3.85 3.39 3 42 3.17 0.39 0.36 0.40 0.33 0.41 0.32 0.34 0.38 0.43 0.38 0.40 0.37 0.78 0.70 0.79 0.71 0.74 0.73 0.62 0.80 0.83 0.77 

## o 78 0.66 38.7 31.4 39.1 33.7 38.0 26.7 39.5 34.3 37.7 34.9 37.6 38.2 0.92 0.80 0.91 0.94 0.96 0.88 0.90 0.94 0.89 0.86 0.90 0.86 0.23 0.19 0.21 0.17 0.17 

## o 16 0.20 0.19 0.24 0.18 0.22 0.19 191.6 188.9 306.3 230 1 275.5 192.2 201·.9 217.8 165.9 197.4 190.8 255.0 30.8 22.5 30.7 32.5 32.2 32.4 28.5 26.7 42.4 29.5 30.2 34.2 212.9 230.9 

## ·ZT4-.-i-­

197.0 183.2 224.7 189.0 190.2 208.5 219. '1 214.3 218.1 CJl J
1,000 51 4.19 0.39 0.92 36.1 0.79 0.21 177.9 21. 2 230.7 52 4.33 0.39 0.86 31. 1 0.87 

## o 22 177.7 33.4 210.4­ 53 4.62 0.40 1. 08 31. 1 0.82 0.17 161.4 31. 6 230.3 54 3.99 

## o 40 0.93 52.1 

## o 96 0.22 193.5 29.2 189.6 55 4.67 0.42 1.00

- 38.3 0.91 0.23 168.2 28.0 203.9 

## Reco;e-r-i------------------------------------------··------------------------------------------------------------------------------------------------------------------­ 56 57 58 

### 59 60 2.93 3.10 2.95 2.92 2.74 0.38 0.44 0.44 0.38 0.39 0.77 0.70 0.88 0.80 0.70 39.6 30.4 36.2 40.4 36.2 0.89 0.73 0.79 0.82 0.B2 0.21 0.23 0.22 0.26 0.23 189.2 184.1 206.9 194.5 153.7 29.3 25.3 25.8 31.4 33.6 210.6 241.0 233.1 234.0 224.4**~** (l)

### BII-0908

Addendum



### Sex 13-1 Twenty-eight-dayrepeated-dose oral toxicity study in rats 

### Pathological findings 

### ofindividual animals Animal Exp.group N Fate Macroscopic findings o. Histopathological findings') 

### 1 ta No abnonnalities detected No abnormalities detected

................... a9

................ "'

.......... 00_

.. ____ ....................................................... ___ .... '"

............. "'

.............. __ ......... "'

......... ___ .' 2 ta No abnonnalities detected No abnonnalities detected**~** .. _ ........................... _ ........................................................................... __ ......................... _ .... e
........................................................ , 3 ta No abnonnalitiesdetected Adrenal Male Vehicle control .•.•..•.

_ .......................
_ •..

_ ..••.•.....•.•...•..•••

### X?c.~l.~~~~l?~y.**.':~ P~r:!~~!** **.~:!~.:**

# .... 4 ta Noabnonnalitiesdetected No abnormalities detected

,. ................................................... _ ..........................._ .... _ ......................... _a.
.... _ ........ "'

................... _ .................. ___ .... __ ..........**~** 5 ta No abnonnalities detected No abnormalities detected a) Organs/tissues examined 

### as follows: trachea, lungs. forestomach, glandular stomach, duodenum,jejunum, ileum, cecum,

colon, rectum, liver, heart, kidneys, urinary bladder, testes, epididymides, prostate, seminal vesicle, cerebrum, cerebellum, 

### pons, spinal cord, sciatic nerve, bone marrow, 

### axillarlymph node, mesenteric lymph node, spleen, thymus, pituitary gland, thyroid, parathyroid, adrenals and eye ball. ta, terminal autopsy. +,slight.



## -196­ 462B11-0908

Addendum 13-2 Twenty-eight-day repeated-dose oral toxicity study in rats Pathological

### findings ofindividual animals Animal 

## Sex Exp.group 

### N Fate Macroscopic findings Histopathological findings') 

### o. 6 ta Noabnormalitiesdetected No abnormalities detected



## ....................................................................................................-

## ............................................................... .,

### .. , 

### 7 ta No abnormalities detected No abnormalities detected Vehicle control

...................................................................................................... Male 8 ta Noabnormalities detected No abnormalities detected (Recovery) 9 ta Noabnormalities detected No abnormalities detected



## ...............

_ ...

## ............................................ ""

## ............................................... "'

## ..................

# - .................................................................. 10 ta No abnormalitiesdetected No abnormalities detected a) Organs/tissuesexamined

### as follows: liver, kidneys, epididymides, cerebrum, cerebellum and pons, ta, terminal autopsy.



## -197­ 463

### Bll-0908

Addendum 13-3 Twenty-eight-dayrepeated-dose oral toxicity study in 

### rats 

### Pathological findings 

### ofindividual animals Exp.group Animal F Sex 

### ate Macroscopic fIDdings Histopathological findings') (mglkgldax) 

### No. 11 ta 

### No abnormalities detected No abnormalities detected 12 ta 

### No abnolmalities detected Noabnormalities detected 13 ta 

### No abnormalities detected Liver MaJe 25



## .................
_ ._....
_ ....
_ .......
_ ....
_ ..............._.**!.?!:~l. !'~~~~~~~** **~.~~:p.~!,?!.?'!~.;.**

## _.__._.' 14 ta No abnormalities detected No abnormalities detected

.....................

### __..

## -_...............

## _- ...'"........................................................... 

### ...... "'...........

## -.......... ,.... ,...................................**~** 

### 15 ta 

### No abnormalities detected Liver Microgranuloma

### + a) Organs/tissues

### examined as follows: liver. ta, 

### terminal autopsy. +, slight.



## -198­ 464BII-0908

Addendum 13-4 Twenty-eight-dayrepeated-dose oraltoxicity studyinrats Pathological 

### findings ofindividual animals Exp.group Animal Fate Macroscopic fmdings (mgfkglday) No. Sex Histopathological findings&) 16 ta Jejunum Jejunum Diverticuluin Diverticulum Liver

• _ ••••••••••••
__ ••

_••••••••

__••
_ •••••
_ •••••••••••••••••0.0•••

### ~!~~<?~':!!~!,l!~ 

## :!'.•••....••..

__ '_0'.

__• Male 150 17 ta Noabnormalities detected No abnormalities detected 18 ta Noabnormalities detected No abnormalities detected

................. "'...........................................................

## .... "'
....................................................................

## _-............................



### _ .... 19 ta No abnormalities detected Noabnormalities detected 

### 20 ta No abnormalities detected Noabnormalities detected a) Organs/tissuesexamined 

### as follows: liver and macroscopic lesion. ta, terminal autopsy. +, slight.



## -199­ 465B11-0908

Addendum 13-5 Twenty-eight-dayrepeated-dose oral toxicity study in rats Pathological fIndings ofindividual animals Sex Exp.group Animal Fate Macroscopicfmdings Histopathological findings·) {mg/l<Jyday) No. 21 ta Liver No abnonnalities detected Enlargement Liver Kidney No abnonnalities detected Enlargement(bilateral) Kidney No abnonnalities detected 

### 22 ta Kidney

. Liver Enlargement(bilateral) Microgranuloma 

### + Skin Kidney

Loss ofhair (neck) 

### No abnonnalities detected Skin Male 1,000 Scab fonnation 

### +

..................... ,.

### .. "'

## ....

## -.... ,.

### .............

_ 

### ..

# _. '"'.,

### ....-........................-­



### _ ..............................

## -_...............................



### _ ...... ,.

### ............................ 23 ta Kidney Kidney Enlargement (bilateral) Pelvic dilatation, unilateral 

### +



## ..................**~:~~j.S .~i}~!~!!1?!.l.(~i~~9** ••..•••.•......•••..••.•••••.••.•.••••••••••.•.•••
___.•••.•. 

### 24 ta Liver Liver Enlargement Diffuse fatty cbange ofhapatocytes

### + Kidney Kidney

25 ta Kidney No abnormalities detected Enlargement (bilateral) Kidney 

### No abnonnalitiesdetected 

### a) Organs/tissuesexamined as follows: trachea, lungs, forestomach, glandularstomach, duodenum,jejunum, ileum, cecum,

colon,rectum, liver, heart, kidneys, urinary bladder, testes, epididymides, prostate, seminal vesicle, cerebrum, cerebellum, pons, spinalcord, sciaticnerve, bonemarrow, axillarlymphnode, mesenteric lymph node, spleen, thymus,pituitarygland, thyroid, parathyroid, adrenals, eye ball and macroscopic lesion. ta, tenninaJ autopsy. 

### +, slight.



## -200­ 466l 

### BII-0908



### Addendum 13·6 Twenty-eight-dayrepeated-dose oral toxicity study ill 

### rats Pathological fmdings ofindividual animals 

### Exp.group Animal 

### Sex (!P8!kglday) 

### No. Fate Macroscopicfmdings Histopatbological findings'

1,000 Male (Recovery) 26 ta 

### No abnormalitiesdetected 

### Noabnormalities detected

....................................................... "'

...... '" "'

........ 4>

..... _ ...... __ .......................... __ ....... ................ '

.......... ,. ... ".

...................... . 

### 27 

### ta No abnormalities detected 

### No**abnormall~BS**

### detected



# ..................................""

# .............................................

# - ......................................................... ,

## ..............

# --_....... 28 ta 

### No**abn~lities** 

### detected No 

### abnormalities**de~ted** 

# .......................................................""'

# .............................................

# -..............

# -._-................................,.

# ....,.

## ............ 3.0 ta 

### No

### abnormalities detected No abnormalities detected a) Organs/tiSSUes examined as 

### follows: liver, kidneys, epididymides. cerebrum. cerebellum and 

### pons. ta, 

### terminal 

### autopsy.



# -201­ 467

### 1311-0908



### Addendum 13-7 Twenty-eight-day repeated-dose oral toxicity study in rats Pathological ti.ndings ofindividual animals Animal F Sex Exp.group ate Macroscopic findings Histopathological findings') No. 

### 31 

### ta No abnonnafnie[ldetected Noabne>rmalities detected

.................. ""

...... to

...................................__ ............................ __ .......................-.... '!"

................... "'

......... - .............**'"',.~** 

### 32 ta No abnMnalines detected No abnonnalities derected



# ..............................................

# - ................................................._._

# .................

# - ..........................-_

# .............. Female 

### Yehiclecontrol 33 ta 

### No abnormalities detected No abnormalities detected**~**

"

# .................................................................................................-

## ....-

## ........,.

# ...............
-

# ........................-_.......... - 

### 34 fa 

### No abnormalities detected No abnormalities detected



# ........... "

## ............. "'

# ...................

# ........

-

# ..................-.........

# -_..

# - ............................................................. " "**~** 

### 3S 

### ta**No~bnormaJities** 

### detected 

### Noabnormalities detected a) Organs/tissues examined as follows: trachea, lungs,forestomach, glandular stomach, duodenum,jejunum, ileum, cecum,

colon, rectum, liver, heart, kidneys, urinary bladder, ovaries, uterus, vagina, cerebrum, cerebellum, pons, spinal cord, sciaticnerve, bone marrow, axillar Jymph node, mesenteric lymph node, spleen, thymus, pituitary gland, thyroid, parathyroid, adrenals and eye ball. ta, terminal autopsy.



## -202­ 468

### 811-0908

Addendum 

### 13-8 Twenty-eight-dayrepeated-dose 

### oral toxicity study in

### rats Pathological fmdings ofindividual animals Animal F 

### Sex Exp.group N ate Macroscopicfmdings Histopathologicalfindings') 

### o. 36 ta 

### Noabnormalities detected No abnormalities detected

... __ ................................................................................................ __ ........... OJ

......... _ ............ OM

_ ............................ 10'

.... _ .......... . 

### 37 ta 

### No abnormalities detected No abnormalities detected Female Vehicle control (Recovery) 38 ta Noabnormalities detected No abnormalities detected



### ...................
_ 

## .............................................

# ­ ..............................................................""

## ................................................................. 39 ta 

### No abnormalities detected No abnormalitiesdetected



## .........................................."'_

## .....................

# --........................................

_ 

# ......."'­

_ 

## ........................................................................ 40 ta No abnormalities detected No abnormalitiesdetected a)Organs/tissues examined as follows: liver, heart andkidneys. ta, terminal autopsy.



## -203­ 469

### Bll-0908

Addendum 13-9 Twenty-eight-dayrepeated-dose oraltoxicity study 

### in rats Pathological fmdings ofindividual animals Sex Exp.group Animal Fate Macroscopic fmdings Histopathological findingsa) (mg/kglday) No. 

### 41 ta 

### No abnormalities 

### detected No abnormalities detected



### ...........................................................................

_ ........................................................................



### _ ...................."". 

### 42 ta 

### No abnormalities detected Kidney Female 

## ..............................................•.•...........
.**~)!t:!:?:.~r-:~ ~.~!?:!.l!~."t** 

# ............... 25 

### 43 ta No abnormalities detected No abnormalities detected 44 

### ta No abnormalities detected No abnormalities detected 

### 4S ta No abnormalities detected No abnormalities detected a) Organs/tissues examined 

### as follows: liverand

### kidneys. ta. 

### terminal autopsy. 

### +, slight.



## -204­ 470

### 811-0908

Addendum 13·10 Twenty-eight-dayrepeated-dose oral toxicity studyin**rat~** 

### Pathological findings ofindividual animals Sex Exp.group Animal Fate Macroscopic findings Histopathological findings') (mli!<liday) No. 

### 46 ta No abnormalities detected No abnormalities detected

..-."'.......-

### .......... -

_ ..

### -- -......

_ ...

### ----....................................................................................................................

### __ .-..............

## -_.... 47 

### ta Liver No abnormalities detected Enlargement Liver No abnormalities detected

_ 

### ............................-....



### _ .................................................................................................


## _ ....... '

### ............................... ,.,..



### _ 

### ......**~** 

### Female 150 48 ta 

### Glandular stomach Glandular stomach Blackish region ofrnucosa Necrosis offundic mucosa

### +



# ..................**t~.! .~)**  

# .........
___..................._._..............._ ....................... 49 

### ta No abnormalities detected No abnormalities detected 50 

### ta No abnormalities detected No abnormalities detected a) Organs/tissues examined 

### as follows: liver, kidneys and macroscopic lesion. ta, terminal autopsy. +, 

### slight.



## -205­ 471

### B11-0908

Addendum 

### 13-11 Twenty-eight-dayrepeated-dose oral toxicity study in rats Pathological findings ofindividual animals Exp.group Animal Sex Fate Macroscopic findings Histopathological fmdings') (mg!Jcslday) No. 51 ta Liver Liver Enlargement No abnonnalities detected Kidney Kidney

Enlargement (bilateral) Degeneration and necrosis of tubular epithelium with regenerative

. epithelium 

### + .**-~** 

### ... ....................................

_..

## -......................................

## -_..............................................- ....- ..

### -_............................. 52 ta Liver Liver Enlargement Fatty change ofperiportal hepatocytes

### +

.............

# ----_._.................-.....

## -_..........

## -..

_ .............................................................

_ ...

### __

### ........

## -_........

## -_..



### _ ....

## -..

# -............. 53 ta Liver Liver Enlargement Fatty change ofperiportal Kidney hepatocytes

### +

Enlargement (bilateral) Microgranuloma + Female 

### 1,000 Kidney No abnormalities detected

..........................

## -_.............................................

_ ..

## ---_.-........................-........-............

## -_....

_ ..

## -_....

## -_......-......

_ ..

_

........

### -_....

### -_.. , 54 ta Liver Lung Enlargement Focal inflanunation 

### + Kidney Liver Enlargement (bilateral) No abnormalities detected Kidney

No abnormalities detected 55 ta Liver Liver Enlargement No abnormalities detected Kidney Kidney

Enlargement (bilateral) Degeneration and necrosis of tubular epithelium withregenerative epithelium 

### + a) Organs/tissues examined as follows: trachea, lungs, forestomach, glandular stomach, duodenum,jejunum, ileum, cecum,

colon, rectum, liver, heart, kidneys, urinary bladder, ovaries, uterus, vagina, cerebrum,'cerebellum, pons, spinal cord, sciatic nerve, bone marrow, axillar lymph node, mesenteric lymph node, spleen,thymus, pituitary gland, thyroid, parathyroid, adrenals and eyeball. ta, tenninalautopsy. 

### +, slight.



## -206- 472

### BII-0908

Addendum 13-12 Twenty-eight-dayrepeated-dose oral toxicity study in rats Pathological findings ofindividual animals Sex Exp.group (rng/kglday) Animal No. Fate Macroscopic fmdings Histopathological findings')

Female 1,000 (Recovery) 56 ta Noabnonnalities detected No abnormalities detected

........................................

## -_...

## ---_....---

## --_..

## --_..........................

## -_ ..

_.....

_..........

_..................

_ ..

## --_.....

### -_..

## --_......

### -_....

## -_....... 

### 57 ta No abnonnalities detected No abnormalities detected 58 ta 

### No abnormalities detected No abnormalities detected

..........

_ 

### ..

## ------_....

_ ................

_...................

## -_......................

_ ......................

## --_......-­

### ....

## ----_....

_

### ......

## ---_..

_ ..

### ---­

### _._-. 59 ta 

### No abnormalities detected No abnormalities detected 

### 60 ta 

### No abnormalities detected No abnormalities detected a) Organs/tissues examined as follows: liver, heart and kidneys. ta, 

### terminal autopsy.



## -207­ 473811-0908



## HISTOPATHOLOGICALPHOTOS

474

### BII-0908

Photo. ) Liverofa female ratfrom vehicle control group. 

### NonnaJ. No. 33 animal. HE. )( 180.

Photo. 2 Liverofa female ratfrom 1,000mglkglday group. Fattychange ofperiportaJbepatocytes. No. 53 animal. HE. )()80.

-1­


### 475

### BII..(l908

Photo. 3 Kidneyofafemale rat

### trom vehicle control group. Normal. No. 33 animal. HE. x180.

Photo.4 Kidney

### of.femalerat trom 

### J,000mglkglday

### group. Degeneration and

### necrosis oftubularepitheliumwith

### regenerative epithelium. No. 51 animal. HE. x180.



## -2­
476

### Bll-0908



## PHYSICOCHEMICALREPORT

477

# cERJ

ReceiptNo. 827-08-D-3387

STUDY CODE: XIS-0908



# FINALREPORT

STABILITY ANALYSIS OFKarenzMT (MixtureofPEMB andPE3MB), HOMOGENEITY, STABILITYAND CONCENTRATIONANALYSES OFTHE TEST SUBSTANCEFORMULATION

March 

### 2009



## Hila Laboratory Chemicals Evaluation andResearch Institute, Japan

-1­ 

### 478

### XlS-0908



## STATEMENT

TITLEOF STADY Stability Analysis of KareniMT (Mixture of PEMB and PE3MB), Homogeneity, Stability and Concentration Analyses ofthe Test Substance Formulation (Study Code: 

### X1S-090S)

I, the undersigned, hereby declare that this report provides a correct English translation ofthe final report (StudyCode: 

### X18-0908, issuedon 

### March 

### 16, 2009).

Yuji Kusune, M.S. HitaLaboratory Chemicals Evaluationand Research Institute, Japan



### -ii479

### XlS-0908



### GLP STATEMENT
Hita Laboratory Chemicals Evaluation and Research Institute, Japan**~Q.,-,r:** 

### __-",S=h","ow.:..:.=a""D=e=n=ko",-,,-,K=.K=. Title: Stability Analysis ofKarenzMT (Mixture ofPEMB and_**PE3~B),** 

### Homogeneity, Stability and Concentration Analyses oftheTest Substance Formulation Study Code: X18-0908

I, 

### the undersigned, hereby declare that this study was conducted in compliance with "Concerning Standard of the Testing Facilities Conducting the Test Relating to the New Chemical Substances" on Japanese GLP (Notification No. 1121003 ofthe 

### Phannaceutical and Food Safety Bureau, MHLW, No.3 (November J

### 7, 2003) ofthe Manufacturing Industries Bureau, MET! & 

### No. 031121004 oftheEnvironmental Health Department, MOE (November 21, 2003)).

I 

### also confinned that this

### report accurately reflected the raw data and the test data were valid.

StudyDirector: Sisned in original March 16,2009 Yuji Kusune, M.S.

111­ 

### 480

### XIS-0908

QUALITY**ASSl.JRA.~CE** 

### STATEMENT

Hita Laboratory Chemicals Evaluation and Research Institute, Japan
Sponsor: Shows Denko K.K. Title: Stability Analysis ofKarenzMT (Mixture ofPEMB and**PE3fvtID,-Ho!Il~!tce.ity""** 

### Stabilityand Concentration Analyses ofthe Test SubstanceFormulation Study Code: X 

### 18-0908

This study was inspected by Quality Assurance Unit ofHita Laboratory, Chemicals Evaluation and Research Institute, Japan. The dates inspected and the dates reported 

### these results to the study director and management are as follows

Phasc Date oflnspection DateReportedto Study Director and Management Protocol October 10, 2008 October 10, 2008 IR Spectrum ofTest Substance October]4, 2008 October 14,2008 Homogeneity and Stability Analyses ofTest Substance Formulation October 14, 2008 October 14, 2008
Confirmation ofthe Answer from Study Director (Protocol) October 17,2008 October 17,2008
Amendment to 

### Protocol October 27, 2008 October 27, 2008 Concentration Analysis ofTest Substance Formulation November 

### 6, 2008 November 

### 6, 2008 Amendment to Protocol (No.2) November 10, 2008 November 10, 2008 Raw Data and Draft Final Report February 24, 2009 February 24, 2009 Reinspection ofRaw Data and Draft Final Report February 27, 2009 February 27, 2009 Amendmentto Protocol (No.3) March 12, 2009 March 12, 2009 DraftFinal Report (Second Time) March 12, 2009 March 12, 2009 Final Report March 16, 2009 March 16, 2009

I, 

### the undersigned, hereby declarethat this report provides an accurate description ofthe methods and procedures used in this study, and that the reported results accurately reflect obtained raw data.

Head, QualityAssurance 

### Unit: Signed in original March 16, 2009 Ryuichiro Mizuguchi, D.S.

-iv

### 481

### X18-0908

TABLE 

### 01" CONTENTS 

### Page TITLE

...,............................................................................................................................ 3 SPONSOR

......................................................................................................................... 3 TESTING FACILITY

....................................................................................................... 3 PURPOSE OF STUDy

....·..····....·..................................................................................... 3 GLP COMPLIANCE

........................................................................................................ 3 PERIOD OF STUDY

........................................................................................................ 3 STORAGE AND RETENTION PERIOD OF DATA

........................................................ 4 RETENTION OFORIGINALDOCUMENTS

................................................................ 4 STUDY DIRECTOR AND PERSONS CONCERNED WITHTHE STUDY AND THE OPERATION

.................................................................................................................... 4 APPROVAL BYAUTHOR

.............................................................................................. 4

SUMMARY

...................................................................................................................... 5

MATERIALS

.................. .................................................................................................. 6 J. 

### TEST SUBSTANCE

................................................................................................... 6

METHODS

........................................................................................................................ 7 1. SUBJECT STUDY

..................................................................................................... 7 2. STABILITY ANALYSIS OF THE TEST SUBSTANCE

..................;...................... 7 3. HOMOGENEITY, STABILITY AND CONCENTRATION ANALYSES OF THE TEST SUBSTANCE FORMULATION

.......................................................,............. 8

THE ENVIRONMENTALFACTORS THATMIGHT HAVEAFFECTED THE RELIABILITY OF THE STUDY RESULTS

................................................................. 12

RESULTS AND DISCUSSION

...................................................................................... 12 I. RESULTS

................................................................................................................. 12 2. DISCUSSION

........................................................................................................... 13

-1­ 

### 482

### Xl8-0908

TABLES Homogeneity and stability analyses ofthetest substance formulation 14 2 Concentrationanalysis ofthe doseformulation

........................................................ 14



### FIGURES IRspectrummeasured prior tothe administration period

.............................,........... 15 2 IRspectrum measured afterthe end ofthe administration period

............................ 15



### REFERENCE 1 IRspectrum provided bythe sponsor
....................................................................'" 15



## -2­ 483

### X18-0908

Study Code: X18-0908 Test Substance Code: HR7645 SponsorCode: 

## s-o 170

TITLE Stability Analysis of

### KarenzMT (Mixture of PEMB and PE3MB), Homogeneity, Stability and Concentration Analyses ofthe Test Substance Formulation

SPONSOR ShowaDenko K.K. 13-9, Shiba Daimon l-chome, Minato-ku, Tokyo 105-8518, Japan

TESTINGFACILITY HitaLaboratory Chemicals Evaluation and Research Institute, Japan 822, 3-chome, Ishii-machi, Hita, Oita 877-0061, Japan

PURPOSEOFSTUDY The purpose ofthis study is to determine the stability ofthe test substance during the dosing period, and homogeneity, stability and concentration of the test substance in formulation in "Twenty-Eight-Day Repeated-Dose Oral Toxicity Study ofKarenzMT (Mixture ofPEMB and PE3MB) in

### Rats" (Study Code: B11-0908).

GLP COMPLIANCE This study was conducted in compliance with "Concerning Standard of the Testing Facilities Conducting the Test Relating to the New Chemical Substances" on Japanese GLP (Notification No. 1121003 ofthe Pharmaceutical and Food Safety Bureau, MHLW, No.3 (November 17, 2003) ofthe Manufacturing Industries Bureau, MET! & 

### No. 031121004 ofthe Environmental Health Department, MOE (November21,2003).

PERIOD OFSTUDY CommencementofStudy: October9,2008 Initiation ofExamination (Initiation ofAnalysis); October 14, 2008 Termination ofExamination (Termination of

### Analysis); December26, 2008 Completion ofStudy: March 16, 2009



### -3­ 484

### X18-0908



### STORAGEAND RETENTIONPERIOD OFDATA The raw data, protocol, study contract documents, test substance information, final report and other record documents will beretained in the archive ofthe Hita Laboratory ofour organization for the same period of BII-0908 paper data. After termination of the retention period, any
measures taken win bedone so withthe approval ofthe sponsor.



### RETENTION OF ORIGINALDOCUMENTS An original protocol and an original final report will 

### be retained 

### at HitaLaboratory. The copies oftheir original 

### thatthe study director will be recognized to be accurate copy will be sentto the sponsor.



### STUDY DIRECTOR AND 

### PERSONS CONCERNED WITH THE STUDY 

### AND THE OPERATION Study director: Yuji Kusune, M.S.
Study staff: Masaru Suzuki, B.S. (Analysis ofthetestsubstance)

SaariTakakura, J.L.A.T., JAC.L.C.T. (Preparation ofthetest substance formulation)


## APPROVALBYAUTHOR Studydirector: Signed in oriiinal March 16, 2009 Yuji Kusune, M.S. Analytical Chemistry Section Hita Laboratory



### -4­ 485

### XlS-0908

SUMMARY

The test substance (KarenzMT (Mixture ofPEMB and PE3MB)) was stable during the dosing period ofsubjectstudy (Study Code: B11-0908). The test substance 

### in 10.0 and 0.25 w/v% formulations was stable for 8 days after preparation at 

### cold and dark place and showed good homogeneity. The concentration oftest substance in 10.0, 1.5 and 0.25 w/v% dose formulations for subject study was acceptable level.



### -5 486

### X18-0908

MATERIALS



### 1. TEST SUBSTA.NCE (INFORMATIONPROVIDED BYTHESPONSOR)



### -6­ 487

### XlS-0908



### 1.8 StorageConditions Stored at room temperature (cabinet No. 1 in test substance storage room, tolerance temperature: 10-30°C). Actual temperature between received of test substance to termination of examination was 19.0-22.4°C. 

### It satisfied tolerance. Avoid oxidizing agent. 1.9 

### HandlingPrecaution Glove, mask, cap, goggles and lab coat were worn. The test substance was treated in the draft since the test substance has characteristic odor.



### METHODS

1. SUBJECTSTUDY Twenty-Eight-Day Repeated-Dose Oral Toxicity Study ofKarenzMT (Mixture ofPEMB and PE3MB)

### in Rats (Study Code: B11-0908)



### 2. STABILITYANALYSIS OFmETEST SUBSTANCE The infrared 

### (IR) spectrum was measured by IR spectrophotometer before and after the dosingperiod ofsubject study. 

### 2.1 MeasurementofIR 1) Instrument 

### IR spectrophotometer: FT-720 (HORIBA, Ltd.) 2) Condition Wave number: 4600em']

- 400 cm'l 3) Pre-Treatment Potassium bromide neat 

### 2.2 CriteriaforJudgment IRspectrum ofthe test substance that measured priorto dosing in ourlaboratory should 

### be identical with provided from the sponsor. The test substance was judged to be stable when there are no differences in the

### IR spectrums at beforeand afterdosing period.



### -7 488

### XlS-090S



### 3. HOMOGENEITY, STABILITY AND CONCENTRATION ANALYSES OF THE TESTSUBSTANCEFORMULATION In the homogeneity analysis, the samples were taken (n=l) from the upper, middle and lower layers offormulations immediately after preparation, respectively. These samples

were pretreated and measured (n=l) with high performance liquid chromatography (HPLC). In the stability analysis, the formulations were stored at cold and dark place for 8 days, and the sample was taken (n=l) from the middle layer of the formulations at point of measurement (3 days and 9days after preparation). These samples were pretreated and measured (n=l) with HPLC. In the concentration analysis, the samples were taken (n=1) from the middle layer ofdose formulations immediately after preparation for subject study. These samples were pretreated and measured (n=1) with HPLC. 

### 3.1 TheTest SubstanceFormulation 1) Homogeneity and Stability Analyses (1) Concentration 10.0 and 0.25 

### w/v% (2) Preparation Method Weighed 5.00 g ofthe test substance in a beaker, and mixed with olive oil to make 50 mL, and used as 10.0 

### w/v% formulation. Accurate 2.5 mL of 10.0 

### w/v% formulation was diluted with olive oil to make 100 mL, and used as 0.25 

### w/v% formulation. Vehicle: olive oil (LotNo. 0460YS,Fujimi Pharmaceutical) 2) Concentration Analysis The 10.0, 1.5 and 0.25 w/v% dose formulations at first preparation for subject study
were used. 

### 3.2 Outline ofAnalyticalMethod The analytical method was decided, according to results of validation of the analytical method on non-GLP atthetestfacility. 1) Validation oftheAnalytical Method 

### (1) Preparation for Measurement Sample

a) Standard Stock Solution for Validation oftheAnal)'1ical Method Weighed 0.10187 g of the test substance, dissolved in 2-propanol to make 100 mL, and used this solution as 1

# ° 18.7 J.lg/mL standard stock solution for validation ofthe analytical method. b) Sample for Specificity The standard stock solution for validation ofthe analytical method was diluted with 2-propanol to make 50,9 llg/mL standard solution and 50.9**~lg/mL** 

### -8 489

### Xl8-0908

vehicle-containing standard solution (containing 2 v/v% vehicle (olive oil)). 2-Propanol was used as solvent blank, and vehicle blank (containing 2 v/v% vehicle) was prepared. c) Sample for Linearity The standard stock solution for validation ofthe analytical method was diluted with 2-propanol to make 25.5, 50.9') and 102 IlglmL standard solutions. 

# *I: 

### It was llsed 50.9 J1g/mL standard solution ofsample for specificity. d) Sample for Accuracy and Repeatability Standard solutions (concentration: 25.5, 50.9 and 102 llg/rnL, each concentration: n=3) were prepared in the same way ofsample for linearity. (2) Specificity Samples for specificity were measured with HPLC. The variation of detection value (total peak area) ofthe test substance between standard solution with and without vehicle was -2.0%. Therefore, it was confirmed that the result of variation satisfied criteria for judgment (within ±5%). In the results of HPLC analysis of solvent blank and vehicle blank, it was confirmed there were no background and interferingpeaks at the elution peak position oftest substance. (3) Linearity Samples for linearity were measured with HPLC. The calibration curve was made by the concentration of the test substance in the horizontal line and the detection value of test substance in the vertical line. The coefficient of correlation ofcalibration curvewhich was obtained from least square was R=0.999. Therefore, it was continued that the result of linearity satisfied criteria for judgment(morethan 0.999). (4) Accuracy and Repeatability Samples for accuracy and repeatability were measured with HPLC. The concentrations ofthe test substance were calculated with regression formula that

was obtained at linearity. Accuracy and repeatability were calculated with these values. Accuracy of25.5 llg/mL standard solution was 0.4, -3.1 and -0.4%. Accuracy of50.9 llg/mL standard solution was -0.4, -1.2 and -1.0%. Accuracy of 102 llg/mL standard solution was 0.0, 0.0 and 1.0%. Repeatability of25.5, 50.9 and 102 IlglmL standard solution was 1.9, 0.4 and 0.6%, respectively. It 

### was confirmed that the result ofaccuracy and repeatability satisfied criteria for judgment (accuracy: within 

### ±J0%, repeatability: less than 5%).

9 

## 490

### XJ8-0908

2) Preparation for Standard Solution Weighed 0.10113 g ofthe test substance dissolved in 

### 2-propanol to make 100 mL, and used this solution as J011.3 

### J.l.g/mL standard stock solution. Accurate 1 

### mL of standard stock solution diluted with 2-propanol to make 20 mL, and 50.6 JlglmL standard solution was prepared. 3) Pre-Treatment Formulationswere mixed well usinga magnetic stirrer. (1) 10.0 w/v% Formulation (Homogeneity, Stability and Concentration Analyses) Accurate 0.5 mL of fonnulation was dissolved in 2-propano! to make 50 mL. Accurate 

### I 

### mL ofthis solution was diluted with 2-propanol to make 20 mL, and served as an analytical sample (dilution rate: 2000). (2) 1.5 

### w/v% Formulation (ConcentrationAnalysis) Accurate 0.5 mL of formulation was dissolved 

### in 2-propanol to make 30 mL. Accurate 2 mL ofthis solution was diluted with 2-propano! to make 10 mL, and served as an analytical sample (dilution rate: 300). (3) 

### 0.25 w/v% Formulation (Homogeneity, Stability and Concentration Analyses) Accurate 0.5 mL offormulation was dissolved in 2-propanol to make 25 mL, and served asan analytica.1 sample (dilution rate: 50). 4) 

### Conditions ofHPLC Analysis (1) 

### Instruments (HPLC7) Interface: D-7000IF (Hitachi, Ltd.) Pump: L-7100 (Hitachi, Ltd.) Auto sampler: 

# L-noo (Hitachi, Ltd.) 

### LN-VIS detector: L-7400 (Hitachi, Ltd.) Columnoven: L-7300 (Hitachi, Ltd.) Dataprocessor: D-7000 HPLC System manager(Hitachi, Ltd.) (2) Conditions Column: L-columnODS (4.6 mm LD. x 

### 50 mm,CERl) Column temperature: 40°C Mobile phase: 0.5 v/v% phosphoric acid solution/acetonitrile

### == 45/55 (v/v) 

### Flowrate: 1

### mLimin Wavelength: 210nm Injection volume: 10 J.l.L 

### 3.3 DataProcessing 1) Detection Value A total peak area was used as the detection value. 2) Quantitative Analytical Method In validation ofthe analytical method, the result oflinearity was a straight line range
-10­ 

## 491

### X18-0908

of25.5, 50.9 and J

### 02 iJ.gltnL standard solutions, and it passed through the origin ofthe coordinates. Therefore, the concentrations ofanalytical samples were detel1l1ined by single level calibration. 3) 

### Calculation oftheTest Substance Concentration in FOl1l1ulation Concentration oftest substance in each sample (C: 

### w/v%) was calculated with the equation shown below and rounded offto three significantfigures.

CsxA x 

### D C 

### Asx 10000

Cs: Test substance concentration 

### in standard solution (!lg/mL) As: Detection value oftestsubstance in standard solution A: Detection value oftestsubstance in each analytical sample D: Dilution rate in each analytical sample 3.4 CriteriaforJudgment 1) 

### Homogeneity Analysis Thetest substance wasjudged as homogeneous dispersion in vehicle ifa coefficient of variation (CV) was within 

### 5%. The CVwas calculated using the following equation:

CV(%) =:; 

### Standard deviation for concentration oftestsubstancein each 

### x 100 Mean concentration oftest substancein eachlayer

2) Stability Analysis The test substance was judged as stable state in vehicle if 

### a rate to the nominal concentration for the actual concentration 

### (R.N.) and a rate to the mean concentration immediately afterpreparation for the actual concentration 

### (R.P.) were within the range of

### ]OO±10%. The

### R.N. and 

### R.P. werecalculated using the following equation:



### R.N.(%)

### = 

### Actualconcentration 

### x 100 Nominalconcentration

R.P.(%) = 

### Actualconcentration x100 Mean concentration immediately after preparation

3) Concentration Analysis It 

### was confirmed that R.N. was within the range of 100±1O%. The R.N. was calculated usingthe following equation:

-11 

### 492

### X18-0908

R.N.(%) = 

### concentration x 100 Nominalconcentration

ENVIRONMENTALFACTORSTHATMIGHTHAVEAFFECTED RELIABILITY OF STUDYRESULTS

There were no factors thatmight have affected thereliability ofthe study data.

RESULTSAND DISCUSSION

1. RESULTS 

### 1.1 StabilityAnalysis oftheTest Substance IR spectrum oftest substance provided by the sponsor (Reference 1) was identical with that measuredbefore dosing period for subject study (Figure 1). There were no differences in the IR spectra between before and after dosing period (Figures 1,2). 

### 1.2 Homogeneity, Stability and Concentration Analyses of the Test Substance Fonnulation 1) Homogeneity and Stability Analyses Theresults ofhomogeneity and stability analyses ofthe test substance formulation are shown inTable 1. (1) HomogeneityAnalysis CV of 10.0 and 0.25 

### w/v% formulations were 0.6 and 1.0%, respectively. The results satisfied criteria forjudgment. (2) StabilityAnalysis

a) 10.0

### w/v% Formulation At immediately after preparation, R.N. were 101 to 102%. At 3 days afterpreparation,R.N. was 100%, and R.P. was 98.0%. At 9 days after preparation, R.N. was 101%, and R.P. was 99.0%. Allthe results ofR.N. and R.P. satisfied criteriaforjudgment. b) 0.25 

### w/v% Formulation At immediatelyafterpreparation,R.N. were 102 to 104%. At 3 days after preparation, R.N. was 102%, and R.P. was 99.6%. At9 days afterpreparation, R.N. was 102%, and R.P. was 100%. AU the results ofR.N. and R.P. satisfiedcriteriaforjudgment.

-12 

### 493

### Xl8-0908

2) Concentration Analysis The results ofconcentration analysis ofthe test substance formulation are shown in Table2. R.N. of 10.0, 1.5 and 0.25 

### w/v% dose formulations were 99.1 to 101%. All the results satisfied criteria forjudgment.



### 2. DISCUSSION Thetestsubstance was stable duringthedosingperiod ofsubjectstudy. The test substance in 10.0 and 0.25 

### w/v% formulations was stable for 8 days after preparation at cold and dark place and showed good homogeneity. The concentration of test substance in 10.0, 1.5 and 0.25 wlv'% dose formulations for subject study was acceptable leveL



### -13­ 

## 494

### XIS-0908

Homo eneity and stabIlity analyses ofthetest substance ormu atlOn Table 1 

### £ l'
Nominal Time point Layer of Actual R.N. Mean cone.

cone. of measurement cone. (%) (w/v%) (w/v%) measurement 

### (w/v%)
Immediately Upper lO.2 

### 102 after Middle 10.2 

### 102 10.2 preparation Lower 

### 10.1 101


### 10.0 3 days after Middle 10.0 100



# - preparation 9 days after Middle 10.1 101



# - preparation Immediately Upper 0.259 

### 104 after Middle 0.254 102 0.256 preparation Lower 0.256 102

0.25 

### 3 days after Middle 0.255 102


# . preparation 

### 9 days after Middle 0.256 102



# - preparation 

### R.P. CV 

## (%) (%)



# - 0.6

98.0 

# -

99.0 

# -

- 1.0

99.6



# -

100


# - 

# I

I



### R.N.: Rateto thenominal concentration R.P.: Rate totheconcentration measured immediately afterpreparation 

# cv: Coefficientofvariation



### I . C trf I . 

### f th d fi

### ormu atlon 

### Tabl

### e

### 2 oncen a Ion analYSIS 0 

### e ose

Date ofanalysis Nominal cone. (w/v%) Actual cone. 

### (w/v%) R.N. (%) 10.0 9.91 99.1 November 6, 2008 1.5 1.51 101 0.25 0.250 100 R.N.: Rateto thenominal concentration



### -14­ 495

### X18-0908



## ns.2F

__­
..;



### Figure 1 IRspectrum measured priortothe administrationperiod

111.t**r-------------------~**



# , I 0.1001"" to ••• ,., • Ii • •• 1 • •• Q 4600 40lI0 181. tOOO 10lI0 

### coo.0 

### Figure 2 IRspectrum measured afterthe endofthe administration period

. ' •• L-'_----'

1000 400 

### Reference 1 IR spectrumprovided by

### the sponsor
-15­ 

### 496

# CERi

Receipt number 662-08-E-4746 Studynumber 94746



# FINAL REPORT

A 96-hourAcuteToxicityStudyofKarenz MT(Mixture ofPEMB and PE3MB) withMedaka

April 15, 2009

Kurume Laboratory ChemicalsEvaluation and ResearchInstitute,Japan

497

## 94746



# STATEMENT

Kurume Laboratory Chemicals Evaluation and Research Institute, Japan

Sponsor Showa Denko KK



## Title A 96-hourAcuteToxicity StudyofKarenz MT(Mixture ofPEMB and PE3MB) with Medaka

Study number 94746

I, the undersigned, hereby declare that this report provides a correct English translation ofthe FinalReport (Study No. 94746, issued onApril 15, 2009)



# f<..c I

# 7,**~o**

# r Date StudyDirector 

# :lalcJ.l 1ltri1:uuI.U4 Takeshi Matsuura

498

### 94746


# GLPSTATEMENT

Kurume Laboratory ChemicalsEvaluation and ResearchInstitute, Japan

Sponsor Showa Denko 

## KK

Title A 96·hourAcuteToxicity StudyofKarenz MT(Mixture ofPEMB and PE3MB)with Medaka

Studynumber 94746

The study described in this report was conducted in compliance with the following GLP principles:
(1) "Standard Concerning Testing Facility Relating to New Chemical Substances" (November 21, 2003; No. 1121003, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare; November 17, 2003, No.3, Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry; No. 031121004, Environmental PolicyBureau, Ministryofthe Environment) (2) "OECD Principles of Good Laboratory Practice (November 26, 1997, ENV/MC/CHEM (98)17)"

This final report reflects the raw data accurately and ithasbeen confirmed that the test data
arevalid.

Date April 15, 2009 Study Director Signed inQri&inal Takeshi Matsuura

499

### 94746


# QUALITY ASSURANCE STATEMENT

Kurume Laboratory Chemicals Evaluation and Research Institute, Japan

Sponsor Showa Denko 

## KK

Title A 96-hourAcuteToxicity Study ofKarenz MT (Mixture ofPEMB and PE3MB)with Medaka

Studynumber 94746

I assure that the final report accurately describes the test methods and procedures, and that thereported results accurately reflect the raw data ofthe study. The inspections and audit ofthis study were carried out and the results were reported to the StudyDirectorand the TestFacility Management by Quality Assurance Unit as foHows.

Date ofreportto Item ofinspection

## Jaudit Dateofinspection

## Jaudit StudyDirector and Test Facility Management Studyplan draft February 26, 2009 February 26, 2009 Studyplan February 26, 2009 February 26, 2009 

# "---_._--­ Amendment to studyplan March 6, 2009 March 6, 2009 ,-­ 

### March 2, 2009 March 6, 2009 after the exposure Startofthe exposure and March 6, 2009 March 6, 2009**-,~.--,...** 

### .---. i--'-­ 

### April 14, 2009 April 15, 2009 

## Raw data and final reportdraft 

# -_... 

## -. Final report April 15, 2009 April 15, 2009

Date April 15, 2009

Head ofQuality Assurance Unit Signed in original Keiji Shiraishi

500

### 94746



# CONTENTS Page

Title

...................................................................·..········..·····..···......·····..·..,······..

## ·1 Sponsor

......·..·..·..·..······..·· ..·····......··....···....·..·....··......··..·..·..·......····..·······..····..

## ···1 Test facility

......................................................................................................... 1 Objective···

..············..·······..···.............................................................................. 

## 1 Testmethod

........................................................................................................ 1 Applied GLp·

..····..···········..·..···············..·......·······..·········..·······..·········..··....·..··..

## ·1 Dates··

....·······..··..·············..·..······.. ··....·....···........................................................2 Storageoftest item, raw data, etc.·····..······....·....··....·· ..·· ..··....··..·..·....·....·....·....

## ·2 Personnel

....·..··...................................................................................................3 Approval offinal report

....··....·..···......................................................................

## 3 SUMMARY

............................................................................·........·....·..·..·..····4 1. Testitem

.......,.....................................................................·....····..·..··..··....·....·6 2. Test sample

.....................................................................................····.... ····..·....·7 3. Testmaterials and methods

..............................................................................8 4. Results and discussion

...................................................................................... 13 5. Factors that affected reliability oftestresults

..·....···....·..·..........................·..··.. 14

Tables and Figure Table 1 Cumulative mortality Table 2 Observed abnormal response Table 3-1 Dissolvedoxygen concentration oftest solutions Table 3-2 pHoftest solutions Table 3-3 Temperatureoftest solutions Table 4 LCso and LLso to Medaka Figure 1 Concentration-cumulative mortality curve Appendix 1 Chemical characteristics ofdilution water Appendix 2 Analytical method and measured concentration of test item Appendix 3 Calibrationcurve and chromatogram Additional data 1 Resultsofpreliminary studies Additional data 2 Results ofmeasured concentration ofeach component oftest item Additional data 3 Qualitative analysis oftransformed substance

501

### Title

Sponsor

Test facility

Objective

Test method

Applied GLP 94746

A 96-hour Acute Toxicity Study of Karenz MT (Mixture ofPEMB and PE3MB) with Medaka

Showa Denko 

## KK 13-9, Shiba Daimon l-Chome, Minato-ku, Tokyo 105-8518,Japan

Kurume Laboratory ChemicalsEvaluation and Research Institute,Japan 3-2-7 Miyanojin, Kurume-shi, Fukuoka 839-0801, Japan

The objective of this study is to determine the acute effects of the Karenz MT(Mixture ofPEMB and .PE3MB) on fish.

This study was performed according to the following test methods and guidance document. (1) Fish, Acute Toxicity Test stipulated in the "Testing Methods for New Chemical Substances" (November 21, 2003; No. 1121002, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare; November 13, 2003, No.2, Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry; No. 031121002, Environmental Policy Bureau, Ministry of the Environment) (2) OECD Guidelines for Testing of Chemicals, Section 2 : Effects on Biotic Systems, 203 Fish, Acute Toxicity Tcst (Guideline 203, 1992) (3) OECD Guidance Document 23 "Guidance Document on Aquatic Toxicity Testing of Difficult Substance and Mixtures" (September 2000)

This study was conducted in compliance with the following GLP principles: (1) "Standard Concerning Testing Facility Relating to New Chemical Substances" (November 21, 2003; No. 1121003, Pharmaccutical and Food Safety Bureau, Ministry of Health, Labour and Welfare; November 17, 2003, No.3, Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry; No. 031121004, Environmental Policy Bureau, Ministry ofthe Environment) (2) "OECD Principles of Good Laboratory Practice (November 26, 1997, ENV/MC/CHEM(98)17)"



### 502 -1­

## 94746

Dates Study initiation date February 26, 2009 Experimental starting date March 2, 2009 Experimental completion date March 6, 2009 Study completion date April 15, 2009

Storage oftest item, raw data, etc.

(1)

## Test item The test sample'l will be sealed in a storage vessel and stored in archives in this laboratory for ten years after the receipt ofnotice specified under Clause 1

## or Clause 2 in Article 4, Clause 2 or Clause 3 or Clause 8 in Article 4-2, and Clause 2 in Article 5-4 or Clause 2 in Article 24 or Clause 2 in Article 25-3 of "Law Concerning Examination and Regulation ofManufacture, etc. of Chemical Substances". If

## it is not stable for the storage period, it will be stored as long while it is kept stable. Treatment of the item supplied by the sponsor after the storage period will be discussed with sponsor.



# *1 It 

## will be stored as the common sample for storage ofthese studies (Study Nos. 94744,94745 and 94746).
(2)Raw data and materials Raw data, the study plan, documents concerning the study presented by the sponsor, the final report and necessary materials will be stored in archives in this laboratory for ten years after the receipt of the notice specified under Clause 1 or CJause 2 inArticle 4, CJause 2 or Clause 3 orClause 8 in Article4-2, and Clause 2 in Article 5-4 Or Clause 2 in Article 24 or Clause 2 in Article 25-3 of "Law Concerning Examination and Regulation of Manufacture, etc. of Chemical Substances". Treatment of raw data and materials, etc. after the storage period wiU be discussed with sponsor.

503 

## ·2·

### Personnel Study Director

Studypersonal Biology

Analytical chemistry

Approval offinal report Study Director 94746

Takeshi Matsuura Section 4 (Eco-toxicity test area)

Jun Sugimura, Miyuki Narasaki, MankoTanaka, Hiroyuki Togawa Takeshi Matsuura, Makiko Anai Hiroyuki Togawa, Fumiko Kamiaka, Tomoyo Kagamiyama, MarikoTanaka Hitomi Fukui, HiromiAndoh

Date April 15, 2009 Signature Signed in original Takeshi Matsuura

504 

## -3­

## 94746



# SUMMARY

Title

A 96-hour Acute Toxicity Study of Karenz MT (Mixture of PEMB and PE3MB) with Medaka

Testconditions
(1)Test item (2)Test organism (3)Exposure duration (4)Test concentration

(5)Replicate (6)The number oforganisms (7)Dilutionwater (8)Type oftest (9) Preparation oftest solution

(10)Volume oftest solution (11)Temperatureoftest solutions (12)Irradiation condition
(13)Feeding (14)Aeration Karenz MT (Mixture ofPEMB and PE3MB) Medaka (Oryzias lalipes) 

## 96 hours Five exposure levels of 5.0, 3.1, 2.0, 1.2 and 0.8 mglL (a geometric series with a factor of 1.6) as loading rate [respectively 0.51, 0.65, 0.25,0.14 and 0.12 mgILas measured concentration] and control One replicate/test level Seven fish/test level Dechlorinated tapwater Semi-static (renewal at every 24 hours) Test sample and dilution water were mixed to produce each nominal lording rate, and then they
were stirred for 48 hours. They were filtrated to prepare the test solution. About 4.9 Utest level 23.9 to 24.2°C Artificial light ofwhite fluorescent lamp, 16-hourlight/8-hour dark No feeding No aeration (15)Analysis ofconcentrationoftest item in test solution HPLC analysis 

## (At the start of the exposure, before and after the renewal and the endofthe exposure)

505

### 94746

Results
(1)Solubility in dilution water (24±1°C) 10mgfL (measured value in preliminary study)
(2) Concentration oftest item in test solution (Percentage oftheinitial concentration) At the start ofthe exposure and after the renewal Below the lowerdetection limit (0.019mg/L) to0.98 mgfL At the before the renewal and the endofthe exposure Belowthe lowerdetection limit (0.019mg/L) to 0.92mglL (50to 97%)
(3) 96-hourLC50 

### (MedianLethal Concentration) 0.16mgfL
(4) The lowest concentration causing 100%mortality at 96hours 0.25mgfL
(5)Thehighest concentration causing0% mortality at 96 hours 0.12 mgfL [Reference value: 96-hour 

## U50(MedianLethal Loading); 1.4mgIL(nominal loadingrate)]
[Thevaluesof(3), (4), (5) are based on geometricmeanofthe measuredconcentrations.]



## 506 -5­

### 94746



## 1. Test item



# . -------..... -

### .....

## -_.....-._._--------_....

### ----------.

# ..**~---** 

# ..

# -----..

# ----..**-----~-.---** 

# ....-----.-------.----

# ..**-----.~---------.** 

# .

507 

### -6­

## 94746



## 2. Test sample
:-.-

## ...**~.-------** 

## ..---.-.--.-----.--.----

# ...

# -....

# -----....

## _------.......------

### ......

## _------_..

## _------- ......

### -_._-_._._.---._._---;



# . 

# . . 

# .

508 

### -7­

## 94746

2.5 Storage condition and confirmation ofstability at storage condition Storage condition Dark storage place at room temperature and avoided contacting with oxidizing agent. Confirmation ofstability The stability ofthe test item during the test period was confirmed by no alteration in the 

## IR spectra ofthe test item before the experimental start and after the experimental completion.

3. Testmaterials and methods
3.1 Test organism (1) Species Medaka (Oryzias latipes) 

## (2) Reasonfor selection ofspecies This species is recommended in the test guidelines. (3) Size Totallength 2.3 ± 1.2 cm Size oftest organism was applied the regulated size set to test method (1). (4) Supplier Kurume Laboratory, Chemicals Evaluation and Research Institute,Japan 3-2-7 Miyanojin, Kurume-shi, Fukuoka 839-0801,Japan (5) Acclimation Medaka hatched out on November 26, 2008 were acclimated for 24 days by flow-through condition under the same water quality (dechlorinated tap water), temperature (24±1QC), light and dark period (16-hour light/8-hour dark) as test condition (on February 6 - March 2, 2009). The mortality was 0% during the 7 days before the start of the exposure. The test organism at the start of the exposure was 3-month-old fish. The test organisms were not treated with a medicament for external disinfection. The test organisms were fed the feed mixture for carp (2C), and not fed for 24 hours before the start of the exposure. Dissolved oxygen concentration in breeding water during acclimation was kept not less than 80% of air saturation value. A 96-hour acute toxicity test of CUS04 5Hz 

### O 

## (Reagent chemical, Wako Pure Chemical Industries, Ltd.) to confirm reproducibility ofthe test system was carried out on October20

- October 24, 2008, and the 96-hour LCso was 0.34 mglL. This value was within the stipulated range (mean ± 2S.D. : 0.14

- 0.97 mgIL) [mean ± S.D. : 0.56 ± 0.21 mgIL (n=42)J to background data in this laboratory. AU of the values shown above for the reference substancewere convertedinto CUS04 value,

509 -8­

## 94746

(6) Allocation to the test groups Medaka were allocated at random to each test group.
3.2 Dilution water Dechlorinated tap water, aerated sufficiently and controlled temperature, was used. Some chemical characteristics of the dilution water measured regularly are listed in Appendix 1.



## 3.3 Test apparatus and equipment
(1) Test apparatus Test vessel : 7.8 L Glass tank (diameter: 24ern, depth: 28 em) The test vessels were covered with lid in order to prevent dust, and volatilization ofthe test solution.
(2) Test equipment Waterbath Plastictank Warming/coolingunit (TypeHCA250, Sato craft)

3.4 Testconditions
(1) Conditions ofexposure (a) Type oftest The test organismswereexposedto the test solution containingthe test item. The test solutionswere renewed at every 24 hours as semi-static regime.
(b) Exposure duration 96 hours
(c) Test concentration Five exposure levels of 5.0, 3.1, 2.0, 1.2 and 0.8 mg/L (a geometric series with a factor of 1.6) as loading rate were used in the test. The exposure levels and the factor were decided based on the results of preliminary studies. The results ofthe preliminary studies are shown in Additional data 1.


## (d) Control The dilution water without the test item, which was treated in the same stirring manner as the test solution, wasused as the control.

510 -9­

## 94746

(e) Replicates One replicate/test level 

## (f) The number oforganisms Seven fish/test level (g) Volume oftest solution About 4.9 Utest level
(2) Conditions oftest environment
(a) Watertemperature 24±1°C
(b) Dissolvedoxygen concentration The test was performed in the condition where dissolved oxygen concentration was at least 60% or more of the saturated concentration at the watertemperature. Aerationwas not used duringthe exposure. (c) pHThe test wasperformed without adjustingpH.
(d) Irradiation condition Artificial light ofwhite fluorescent lamp, 16-hourlight/8-hourdark
(e) Feeding Testorganisms were not fed during the exposure.
3.5 Preparation oftest solution The test sample and dilution water were mixed to produce each nominal lordingrate and stirred by magnetic stirrer for about 48 hours. The solution was filtrated with a glass fiber filter (ADVANTEC, GB-l40, 0.4 /1-m filter unit) by suction. Mter the filtration, the filtrate was aerated for about one hour under about 24°C in order to
recover decreased dissolved oxygen concentration by suction and then used for test solution.

511 

### -10­

### 94746

3.6 Observation and measurements
(1) Observation oftest organisms Mortality and visible abnormality were observed at 3, 24, 48, 72 and 96 hours after the start of the exposure. A fish was considered as dead if the observable motion (motion ofmouth and opercula etc.) were not observed and touching ofthe caudal pedunclewith glass rod produced no reaction.
(2) Total length and body weight oftest organism The test organisms in the control group were used for measuring total length and bodyweight afterthe end ofthe exposure. (3) Appearance oftest solution Appearance ofthe test solutions was observed at the start and before renewal oftest solutions (24 hours after).
(4) Qualityoftest solutions Dissolved oxygen concentration, pH and water temperature of the test solutions were measured at the start of the exposure, before and after the renewal, and the end ofthe exposure. They were measured also at the time that confirmed all test organisms dead at the exposure level, where all test organisms died at the exposure level. The dissolved oxygen concentration measurements were carried out with an oxygen meter (YSI Incorporated, YSI MODEL 58). The pH measurements were carried out with a pH meter (DKK-TOA, Model HM-21P). The temperature measurements wer:e carried out with a calibrated red alcohol thermometerofglass stick type.
(5) Concentration oftest item in test solution The concentration of the test item in the test solutions was measured at the start ofthe exposure, before and after the renewal, and at the end of the exposure. The measurement was conducted also at the time that confirmed all test organisms dead at the exposure level, where all test organisms died at the exposure level. The test solution was taken out from the middle layer of the test vessel and was used for the measurement. The concentration of the test item was analyzed by high-performance liquid chromatography (HPLC). Analytical method and measured concentration of test item are shown in Appendix 2, and analytical calibrationcurve and chromatograms are shownin Appendix 3.

512 -11­

### 94746

3.7 Calculatingmethod ofLCso'3 and LLso'4 

### Results of this study were estimated using Computer Program (running on Microsoft software "Excel")developed by our laboratory.
(1) LCsoThe results of this study were estimated by geometric mean of the measured concentrations For 48 and 72 hours, the LCsowas calculated by Probit analysis, and the 95% confidence interval andthe slope ofthe dose-response curve were also calculated. The 96-hourLCsowas calculated by Binomialtest. For 24 hours, the LCso was estimated as 

### "> the highest test concentration (0.65 mg/L)" since the mortality over50% was not observed.
(2) LLso (reference) The results ofthis study were estimated by the nominal loading rate For 48, 72 and 96 hours, the LCso was calculated by Binomial test. For 24 hours, the LLso was estimated as 

### "> the highest test concentration (5.0 mglL)" since the mortality over50%was not observed.

*3 LCso (Median Lethal Concentration) :The test item concentration at which 50% of the test organisms causesmortality duringthe exposure.

*4 LLso (Median Lethal Loading) : The test item nominal concentration at which 50% ofthe test organisms causes mortality duringthe exposure.
3.8 Validity ofthe test
(1) Themortality inthe control should not exceed onefish.
(2) Dissolved oxygen concentration must be at least 60% ofthe air saturation value at the water temperature in the test duringexposure.
3.9 Treatment ofnumerical values Values wererounded offin accordance withJIS Z 8401 rule B, 1999. (JIS ;Japanese Industrial Standards)

513 -12­

## 94746

4. Results and discussion

The test concentrations in the following sentence is expressed using geometric mean ofthe measured concentrations which are 0.51,0.65,0.25,0.14 and 0.12 mg/L, which are respectively5.0, 3.1, 2.0, 1.2 and 0.8 mg/Las the loadingrate.

4.1 Mortality Cumulative mortality of each observation time was shown in Table 1. 

## Concentration-cumulative mortality curve is shown in Figure 1. 

## The lowest concentration causing 100% mortality within 96 hours was 0.25 mg/L, and the highest concentration causing no mortality within 96 hours was 0.12 mg/L. Number ofdead fish in the control at the end ofthe exposure was 0, which meets the criterion for the validity ofthe test (Le. not exceed one fish).

4.2 Observed abnonnal response The abnormal responses observed during the exposure are shown inTable 2. No abnonnal responses were obtained in the controL The following results ofobservation were based on the comparison with the control organisms. Observed abnormal responses during the exposure were swimming at the water surface, light body color and reduced activity.

4.3 Size oftest organism [Mean 

## ±Standard deviation (n=7)] Total length 2.5 ±

## 0.076 em Body weight 0.13 ± 

## 0.011 g
4.4 Observation and measurement oftest solution
(1) Appearance oftestsolution The test solutions were clear and colorless at the start ofthe exposure. The appearance kept until before the renewal.
(2) Quality oftest solutions Qualities ofthe test solutions are shown in Tables 3-1, 3-2 and 3-3. The measured values ofdissolved oxygen concentration, pH and temperature during the exposure ranged from 7.2 to 8.4 mglL, 7.5 to 7.9 and 23.9 to 24.2°C, respectively. The measured values of dissolved oxygen concentration met the criterion for the study validity (at least 60% or more of saturate concentration'5 

## at the watertemperature).

*5 Saturated dissolved oxygen concentration (23 to 25°C): 8.39 to 8.11 mg/L (JIS K 0102, 2008)

514 -13­

## 94746

(3) Concentration oftest item in test solution The results of the measured concentrations of the test item are shown in Appendix 2. The measured concentrations of the test item in the test solution were from below the lower detection limit (0.019 mglL) to 0.98 mglL at the start of the exposure and after the renewal, and from below the lower detection limit (0.019 mglL) to 0.92 mglL (50 to 97% of the initial measured concentration) before the renewal and the end of the exposure containing with the time all 

## test organisms died.

4.5 LC50

## and LLso The LCsos and LLsos at each observation time are shown in Table 4. 

## The 48-hour and 96-hour LCso were respectively 0.49 mgIL (95% confidence interval: 0.012 to 0.60 mglL) and 0.16mglL. As reference value, the 48-hour and 96-hour LLsoestimated based on the loading rate were respectively 2.6 mgIL and 1.4 mgIL.
4.6 Discussion This study was conducted as a test in order to estimate LC50 

## ofthe test item for the test organisms below the solubility ofthe test item in dilution water. As a result, the 96-hour LCso based on the measured concentration was 0.16 mglL, the 96-hour LCso based on thenominal loadingrate was 1.4mglL. The concentration-response ofcumulative mortality was not linear between 3.1 and 5.0 mglL. However, it was confirmed that the measured concentration in the test solution during the exposure was also not linear at the same exposure levels. Therefore, the concentration-response ofmortality was linear. Since the cause ofthe reversal of the test item concentration was the reversal ofthe PEMB 

## concentration, it
was highly likely that PEMB 

## concentration was involved in the effect on organism greatly (reference: Additional data 2). It 

## was considered that expected causes of reversal of the concentration were to be poorly water-soluble for the test item, and to adopt the method to prepare the test solutions each exposure level. In addition, because it was confirmed that the converted products derived from the test item produced in the test solution during exposure, there was a possibility that the test item concentration was affected by productive condition of the converted products (reference: Additional data 3). Since it was confirmed that the maintenance ratio ofthe test item concentration in 

## the test solution varied, the definitive study was conducted the renewal as much as possible. The environmental conditions were within the suitable range, therefore, it is concluded that this studycompliedwith the applied test guidelines.



## 5. Factors that affected reliabilityoftest results
There were no factors which might have affected the reliability ofthe test.
515 

## -14­

### 94746

Table 1 Cumulative mortality

Measured Cumulative mortality (%) concentration*6 

### (mglL) 3 hours 24 hours 48 hours 72 hours 96 hours
Control 0 0 0 0 

## 0

0.12 0 0 0 0 0
0.14 0 0 0 0 29
0.25 0 0 0 86 100
0.65 0 14 86 100 100
0.51 0 0 57 86 100 *6 Geometric mean of measured concentration (also expressed as measured concentration in the following tables and figure)

Table 2 Observed abnormal response

Measured concentration (mglL) Result ofobservation (Left column: Numberofaffected fish 

## ITotal survival number, Right column: Symptom detail) 3 hours 24 hours 48 hours 72 hours 96hours Control 

## on 1 N 

### I 017 [ N 

## on 1 N j 

## on

# I N 

## on

# I N 

### 0.12 I 

## on N 

# on! N 

## on N I 

## on 

## I N 

## on N 

### 0.14 

## on N 

## on N 

## on 

# ::::Ei N

N 0/1 : N 

### ! 0/5 N 

### 0.25 

## on N 

## on, N 

## on


## - 

# - 0.65 

## on ' N 0/6 N III AS(l) 

# - 1 

### ! 

# - 

# - - 0.51 0/7 N 0/7 N 113 A1S(1} LT(I) RA(l) 

## on N 

# - - N: Normal (No abnormal response) Value in parentheses expresses the number ofindividuals shown the symptom.
- :All test organisms died. Abbreviation ofsymptoms AS :Swimming at the surface LT:Light body color RA :Reduced activity

516 

### -15­

### 94746

Table 3-1 Dissolved oxygen concentration oftest solutions

Measured 24hours 48hours 72 hours 

# I concentration At the Atthe slart Beforethe After the Beforethe Afterthe Deforethe After the 

### end (mglL) renewal renewal renewal renewal renewal renewal 

### Control 

## 8.3 7.7 8.4 7.8 8.2 7.8 8.4 7.9

0.12 7.8 7.6 8.4 7.7 8.1 7.8 8.3 7.8 0.14 8.1 7.6 8.4 7.6 

## 8.1 7.6 8.2 7.7 0.25 8.2 7.5 8.3 7.6 8.1 7.7 8.1 8.1* 0.65 8.1 7.4 8.3 7.2 8.2 8.1* 

# - - 0.51 

### 8.2 7.6 8.3 7.5 8.2 8.0 8.2 8.2* Unit: mgIL
- :All test organisms died. 

# * : It

### indicates the measured value at the time that confirmed all test organisms dead.

Table 3-2 pHoftest solutions

Measured concentration (mglL) At the start 24 hours 48 hours 72hours Atthe end Before the renewal After the renewal Beforethe renewal After the renewal Before the renewal After the renewal 

### Control 7.7 7.5 7.7 7.6 7.7 7.5 7.7 7.5

0.12 7.9 7.5 I 

### 7.9 7.5 7.9 7.5 7.9 7.5 0.14 7.9 7.5 7.9 7.5 7.9 7.5 7.9 7.5 0.25 7.9 7.5 7.9 7.5 7.9 7.5 7.9 7.8* 0.65 7.9 7.5 7.9 7.5 7.9 7.7* 

# - 

# - 0.51 7.9 7.5 7.9 7.5 7.9 7.7 7.9 7.8*

- : All test organisms died. 

## * : It

### indicates the measuredvalue at the time that confirmed all test organisms dead.

517 -16­

### 94746

Table

## 3-3 Temperature oftest solutions

Measured 

# 'A h

. I 

## tt e concentratIOn (mglL) : start 24 hQurs 48hours 72 hours Atthe end Before the renewal Aftertbe renewal Defore the renewal Afterlhe renewal Before the renewal Afterlbe renewal 

### Control 23.9 24.0 24.0 24.0 24.1 24.0 24.1 24.0

0.12 23.9 24.0 24.0 24.1 24.2 24.0 24.1 24.0
0.14 24.0 24.0 24.0 24.2 24.2 24.1 24.1 24.0
0.25 24.0 24.0 24.0 24.1 24.2 24.0 24.1 24.0* 0.65 24.0 24.0 24.0 24.2 24.2 24.0·


# - 

# - 0.51 24.0 24.0 24.0 24.1 24.1 24.0 24.1 24.1* Unit: °C
- :All test organismsdied. 

### '" : Itindicates the measured value at the time that confirmed all test organisms dead.

Table4 LCsoand LLso to Medaka

Exposure duration LCso (mgIL) 95% confidence intervals (mgIL) (Slope ofthe dose-response curve) Statistical procedure 

## !

used for determination ofLCso Referencevalues Statistical LLso'7 

### procedure used for (mglL) detennination ofLLso 24 hours >0.65 

# - (



# -) 

# - >5.0 

# -

48hours 0.49 0.012to 0.60 (9.4) Probit analysis 2.6 Binomial test
72hours 0.23 0.16to0.32 (5.1) Probit analysis 

## 1.7 Binomial test
96hours 0.16 Binomial test 1.4 Binomial test
-: Not obtained *7 Based on loadingrate

-17­ 518

## 94746

100

80**~ ~**  '-' >. 

### 60 

# - .,... 

# "3

.... 0 

# e**~** 

# ·5 ..!:l :::I 

### 40 

# e :::I U



### 20



## o 

# .
. .; 

# ....................•



# . 

# .. . 

# .

-G--24hour**~48hour** 

### ...•... 72 hour



## ---.- 96 hour



### 0.1 1 

# Measured concentration (mgfL)

Figure 1 Concentration-cumulativemortality curve

519 

## -18­

## 94746



# Appendix 1

Chemical characteristics ofdilution water



### 520

## 94746

Chemical characteristics ofdilution water (Sampling on January 14,2009) ResulCs Parameter Unit 

### Lowerlimil

## ordeterminalion Total hardness (as CaC03) 

### 1 

### 42 mg/L < ] 

### 1 mg/L Suspended solid



## - 7.9 (22"C) 

### pH



## - Total organiccarbon mg/L 0.1 0.4 Chemical oxygen demand mg/L 0.6 0.5 < 0.02 Residual chlorine mg/L 0.02 Ammonium ion mg/L < 0.01 0.01 Total cyan mg/L < 0.01 0.01 Alkalinity mg/L 1 

### 29 Electric conductivity mS/m 17.1 

# - Organicphospborous mg/L < 0.1 0.1 Alkylmercury < 0.0005 mg/L 0.0005 Mercury mg/L < 0.0005 0.0005 Cadmium mg/L < 0.001 0.001 Chromium 

### (VI) mg/L <0.01 0.01 Lead mg/L < 0.005 0.005 Arsenic < 0.001 mg/L 0.001 BOTOn mg/L 0.08 0.01 Fluorine mg/L < 0.1 0.1 Iron mg/L < 0.01 

# om Copper mglL < 0.005 0.005 Cobalt 

### < 0.001 mg/L 0.001 Manganese mg/L < 0.01 0.01 Zinc mg/L 

### < 0.005 0.005 Aluminum mglL 

### OJ)42 0.001 Nickel mg/L < 0.001 0.001 Silver mg/L < 0.0001 0.0001 Sulfate ion mg/L 20.1 0.1 Chloride ion mg/L 

### 12 1 Sodium mg/L 

### 14.9 0.01 Potassium mg/L 

### 1.8 0.01 Calcium mg/L 

### 11.5 0.01 Magnesium mg/L 

### 3.3 0.01 1,2-dichloropropane mg/L <0.0001 

### O.[)OOl Chlol'Othalonil mg/L < 0.0001 0.0001 Propyzamide mglL 0.0001 <0.0001 Chlornitrofen mg/L 

### < 0.0001 0.0001 Simazine mglL < 0.0001 0.0001 Tbiobencarb mg/L 0.0001 

### < 0.0001 Diazinon mg/L < 0.0001 0.0001 Isoxathion mg/L < 0.0001 0.0001 Fenitrolhioll mg/L < 0.0001 0.0001 EPN mg/L 

### < 0.0001 0.0001 Dichlorvos mg/L < 0.0001 0.0001 Iprobenfos mg/L < 0.0001 0.0001 PCB mg/L < 0.0005 0.0005



### 521 

### -1 Appendix )­

## 94746



# Appendix 2

Analytical method and measuredconcentration oftest item



## 522

### 94746

1. Pretreatment oftest solution

The test solutions sampled were pretreated according to the following flow scheme to prepare the sample for high-performanceliquid chromatography (HPLC) analysis.

Flow scheme 

# I Test solution 100 mL 

# I

+-Chloroform 30mL(graduated cylinder)
• Shaking (10 min) 

# I 

# I Water layer 

# I I Chloroform layer 

# I

• Transferred chloroform layer to the eggplantflask with

a beaker
• Dryness (rotary evaporator, about40°C, nitrogen stream) +-Acetonitrile/ water"l (111 v/v) about 1mL(pasteurpipette)

• Ultrasonic irradiation (ultrasonic cleaner, about 30 sec)
• Fillingup to2 mL [acetonitrile/ water"l (1/1 v/v),

volumetric flask] 

# I Sample for HPLC analysis 

# I

*1 Tap waterwastreated with ultrapure water system.



## 523 -1 Appendix 2­

### 94746

2. Method ofanalysis

The pretreated samples for HPLC analysis were quantitatively analyzed by HPLC under the following conditions to determine the concentration of the test item. The test item was detected as two peaks on the chromatogram. Therefore, these were analyzed as the test item. The concentration ofthe test item in each sample for HPLC analysis was determined on the basis of a comparison of the total peak area on the chromatogram of the sample solution with that of a standard solution. Some chromatograms obtained are shown in Appendix 3. 

### Analytical conditions Instrument High-performance liquid chromatograph SHIMADZU LC-2010A (Equipped with UV-VIS detector) Column L-column ODS 

### (15 cm 

## x 2.1 mm I.D., Chemicals Evaluation and

Research Institute) Column temp. 40°C Eluent A: Acetonitrile I

### phosphoric acid (1000/1 

### v/v) B: 

### Water"l 

## Iphosphoric acid (1000/1 

## v/v)

Gradient condition Time (min) 

## A(%) B(%) 0.00 50 	 50 20.00 

## 100 

# o 40.00 

## 100 

# o Flowrate 0.2 mIJmin Wave length 210nm Injection volume lO.uL 

### Detectoroutput 2V/AU

3. Preparation ofstandard solution

The standard solution to determine the concentration ofthe test item in the sample for HPLC analysis was prepared as follows. Test sample of 100 mg was precisely weighed with an electronic balance and dissolved in acetonitrile to obtain 1000 mgIL solution of the test item. The solution
was diluted with acetonitrile/ water"1 

## (111 v/v) to prepare 50mglLofstandard solution.

524 -2 Appendix 2­

### 94746



## 4. Calibration curve
The standard solutions of 5.0, 25, 50 and 100 mgIL were prepared for 

## HPLC analysis by the same procedure as described in section 3. These soiutions were analyzed according to the quantitative analytical conditions described 

## in section 2. A calibration curve was drawn from the relationship between the concentrations of standard solution and the total peak area on the chromatogram, and the determination
was confirmed. The calibration curve is shown in Appendix 3. The determination limit ofthe test item was the lowest concentration ofthe standard solution (5.0 mglL) within the calibration range. Therefore, the determination limit ofthe test item in the test solution was 0.11 mglLin consideration ofpretreatment.

5. Recovery test and blank test

5.1 Method The recovery test was conducted by adding the test item (prepared with acetonitrile) to dechlorinated tap water according to pretreatment of test solution described in section 1. The blank test was also conducted using dechlorinated tap water (added acetonitrile) without the test item in the same way as the recovery test. The recovery test and the blanktest were performed in duplicate.

Amount ofthe test item added 

## 100Itg

5.2 Result As a result of analysis by the method of section 5.1, no peak of the test item appeared in the chromatogram of the blank test. Two individual recovery rates and their average on the analytical procedure are shown below. The average ofrecovery rate was used as correction factor for the determination ofthe test item concentrations in the test solutions.
Recovery rate for pretreatment 87%, 89% average 88%



## 525 -3 Appendix 2­

### 94746

6. Results ofmeasurement
The results ofthe measured concentrations of the test item in the test solution are shown below. Appendix table 2-1 Measured concentrations oftest item in test solutions

Measured concentration (mg/L) Nominal (Percentage ofmeasured concentration versus that at each preparation %) concentration (mglL) Atthe start
Control n.d.

0.8 0.16

1.2 

### n.d:3



### 2.0 n.d:4



### 3.1 0.69

5.0 0.51 24 hours old new n.d. n.d.

n.d:4 

### 0.17

n.d."3 0.25

n.d."3 0.61
0.59 0.74 (86) 0.39 0.92 (75) 

# G hOUrs 

### 48 hours old new new n.d. n.d. n.d. n.d.

'4 

### 0.22 0.11 0.19 n.d. (53) 0.22 0.41 0.25 0.35 (85) (61) 0.39 0.65 0.37 0.63 (64) (57) 0.54 0.72 0.65'2

- 

### (73) (91) 0.89 0.26 0.13 0.98 (97) (50) At the Geometric end mean
n.d. 

# /

0.12 0.12 (65) 0.21 0.14 (61) 0.46"2 0.25

(74)



# - 0.65
0.92*2 0.51

(94) old : before the renewal
new : afterthe renewal n.d. ; < 

### 0.11 mgIL
- : It 

### shows no measurementbecause all test organisms died. *2 It 

### indicates the measured value at the time that confirmed all test organisms dead. *3 In accordance with DECO Guidance Document No.23, the detection limit (0.019 mgIL)
was adopted for the calculation of the geometric mean, because the test item was not detected. *4 In accordance with 

## DECD Guidance Document No.23, halfthe determination limit of the test item (0.055 mgIL) was adopted for the calculation ofgeometric mean, because the test item was detected but not quantified.

526 -4 Appendix 2­

## 94746



# Appendix 3

Calibration curve and chromatogram



## 527

## 94746

900000

# ~ C\1 

## 450000

## ~**~**

### ~



# oo 50 

## 100 Concentration (mgIL)

y 

### = 

## 8519x r

# =1.00

Concentration 

### Peakarea (mgIL) 

### (}.tV'sec)



## 5.0 41878 25 212165 50 429373 100 850423

Appendix figure 3-1 Calibration curve oftest item for analysis by

## HPLC



## 528 -1 Appendix 3­

## --- 94746

Study No. 94746 

## l!:tt;LILmcLL

___.
__.
_____.. Operating Date: Mar.02,2009 PlCNO Time Area lieI cht Cone. Cain) ().IV·seo) ().I V)**(~)** I 8.68 61153 5309 14.60 2 15.22 361357 39470 85.4Q



### - Total 423110 100.00

I 

# _L_.

__

# ._JJ

.­



# . I 

## • 0.0 10.0 20. 0 30.0 Operator: Hiroyuki Togawa 

### 94746LOh/Control

-

.. Operatinit Date : liar.02.2009 PKNO Time Area Height Cone. (min) ( pV ·sec) ().IV)**~)**



## n.d. n.d.

# ~ 

# i**~- --"~--**  

## .. ,.---_. 0.0 10'.0 20.0 

### 30. Operator: Hlroy"'l Togawa



### 94746/0hLO.8 mg/l Operatinr; Date : liar.02.2009

..**-~"-~-- -",'~~"-<..----.**  PKNO Time Area Height CoIm. (pain) (ltY·sec) ().IV)**~)** 1 8.68 800B 799 13.51 2 Iii. 23 51846 4344 86.49

Total 59943 100.00



## !. I 

### .1



# . , 

# . 110 10.0 20.0 30.. Operator : HIroyuki TOlal!a


### Date: Uar.4.2009



### Appendix figure 3-2-1 HPLC chromatograms at startofexposure



## 529 -2 Appendix3­

## 94746

Study No 94746 Oparatinr Date

. ller.02.2009 

### 94746iOhLl.2 

### mgiL PKNO Tim. Area fieleht Cone. jilln) (,U'l'680) (pV) (\)



### n.d. n.d.

# ~

# ~**-"----"'_~** __...J..**-~** 0.'0 

### ;0:0 20.0 ___**~'** __0

__••
_ ..... 30.0 Operator : Hiroyuki TOlaR 

### 94146/9.bfl.<LM

__________. 

### ----. ..­ 

### PKNO Operatiol Date: Mar. 02, 2009 Time Area Heicht Cone. (min) (J.l V·sec) (IlV) (t.)



### n.d. n.d.

# ~

# ~**---1~** __.
__....,A.



# . 10.0 0.0 

### ,**,~-"** 20.0 

# ---_

.


### , 311.(1 Operator ; Hiroyuki T ogawa**94146/0hLlilJ!&lb...~** 

### ______ ..
___.
__.__ Operating !late: Iler.02.2009 PKNO

1 2 Time (min) 8.10 15.25 Area (jlV'sec) 93175 162954 Height (/IV) 8671 15911 Cone. <") 36.38 63.62 Total 256129 100.00



# 1 

# 1 1..



### -
....**-~----~"-** . 

# . , 

# . C1.0 10.' 

# ao :!fl. Operator : Hiroyuki Toga"a FIg



### Appendix figure 3-2-2 HPLCchromatograms at startofexposure

530 -3 Appendix)­

## 94746



## Study No. 94746 94746/0h/5.0 mgjL Operating Date : liar.02,2009 /'KilO T i.& Area He iIIht Cone, emin) (1/Nee) (I/V) (I) 8.68 104447 9705 54.88**.1.~.** 23 85859 7627 45. 12 Total 1911306 100.00

Operator : HiroyukI 10ga1lll 

### Fig 

### Date: Mar. 4, 2009 N8IIIe:

## H~rQluh**I~Gwg ,~** **~a** 

## ..!;**~.<,/**



## Appendix figure 3-2-3 HPLCchromatogram at startofexposure

531 

## -4Appendix 3­

## 94746

Study No 94746 std. 50 m&Ll

__.._ .·4.

___ ___ Oparatinll Data: Mar.03.2009**~~·** 

### PKNO Time Area 

### Heiiht Cone. Ullin) (SlY,sIIe) (StV) 0') 1 8.72 62228 5300 14.57 2 

## 1!.f8 364889 39552 85.43 Total 427117 100.00

I

........... A



# . 

# . 0.0 10.0



### 94146[Z4hLControllMO I 

### 20.0 3110



# ---- ­**~-** ---.¥.---­ PKNO Operator: HiIOyuki Togawa Operating Date : liar.03.2009 Til1lll Ana Height Cone. (mi n) (I!V'sec) (SIV) (Il



## n.d. n.d. 

# i**~ ~** 



# . 0.0 10.0 20.0 

### 94146/24h/O. 8**m,!.vMllI~** 

## __...._.**---~** 30.0



### - I'lOO Operator: HiroYllki Togawa Operating Dab : Mar.03.2009 Time 

### Area Height Conc. (min) (IIY·sec) (flV) (S)



## n.d. n.d.

# ~**~ --~---------**  ",.**.----~.---** 0.'0 

### Hi.a 

## to.0 

## JOo Operator : HIroyuki Togalla Fig



## Appendix figure 3-3-1 HPLCchromatogramsbeforerenewal at 24 hours



## 532 -5 Appendix 3­

### 94746



## Study 

## No. 94746 Operating Date: Mar.04.2009**94746/24hLLUgf~*1l\L** 

### .. m**~"""~"** ..-- ... PKNO 

### Time Area Height Cone. (min) (IlV·sec) (pV) (I)



### n.d. n.d.

# ~**~** ......A.---.

____•.

___.-A--

...
________.....



## .. I 300 

### 0:0 Hi.0 20.0 Operator : Hiroyuki Togalla Operating Date: Mar. 04.2009 

### 94**H~Lf5bLLQ** 

## ..lJlW.jt!*liI

..

### ---.--­**~--** Pl(NO Time Area Hei,tit Corn;. (min) (IlV·seo) (/IV) (I)



### n.d. n.d.

# ~ w.

_______**~.---""'--.----"--** Co ,0. 0**94746124bn.L!!l&LLj~** 2!UI**.-~** .. 31).

PKNO 1 2 Operator : 111 royuki TogaWi Operating Date: Mar.04. 2009 tillie Area Heieht Cone. (min) (/IY·aee) (IlV) <l) 8.72 87316 7650 39.33 HU8 134617 12606

_ 60.61 Total 221993 100.00

2

### ~_~ 

# __....L<­

__.

I '11.11 U).O**2O~0** 

# - 30.0 Operator: Hiroyukl Togawa FIg



### Appendix figure 3-3-2 HPLCchromatogramsbeforerenewal at 24 hours

533 -6 Appendix 3­

### 94746



## Study No. 94746 Operatinl Date : Mar. ()4, 2009 PKNO Tillie Arell Height Cone. (ml rll (pV-sec) (pV) (" 1 8,73 818.41 7395 56.50 2 15.25 63010 5063 43.50 TotaI '44951 100.00



### 30.0 Operator: Hiroyukl Togawa 

### Fig Date Mar. 4. 2009 Name

### :H\Yop,l~; TOdAWo.**/g..~**

## ~-1,)

......!.\,..'



### Appendix figure 3-3-3 HPLCchromatogrambeforerenewal at 24hours

534 -7Appendix]­

## 94746



# Additional data 1

Results ofpreliminary studies

535

### 94746

1. Solubility oftest item in dilution water

It 

### was expected that the solubility of the test item in dilution water was below 100 mg/L, therefore, the preliminary studyfor measurement ofthe solubility ofthe test item in dilution water was conducted.

1.1 Measurement ofwater solubilityoftest item
(1)Method The test sample and dilution water were mixed to prepare about 100 mgIL and they were stirred for 24 and 48 hours under the test condition (24±1QC). After that the mixture was left at rest for about one hour, the middle layer was collected and filtrated" with a glass fiber filter (ADV ANTEC, GB-140, 0.4 11m 

### filter unit). The filtrate was analyzed with high-performance liquid chromatography (HPLC) as described in Appendix 2. 

# * Test solution was filtered after the filter was washed with about 100mLofthe test solution.
(2)Result Sample name Measuredconcentration (mglL) Arithmetic mean (mglL) Stirringfor 24 hours Stirringfor48hours Sample-l 10 

# -----­

10 Sample-2 

## 10 

# -------­ Sample-3 10 

# -------­ Sample-4 

# ----­ 10 

### Sample-5 

# -------­ 11 

### Sample-6 

# -----­ 9.2

1.2 Summary It was decided that the solubility ofthe test item in dilution water was 10 mg/L, which was the average ofmeasured concentration stirred for 24hours and 48hours.

536 -1 Additional data 1­

### 94746

2. Effectontest organism

2.1 Preliminary study
(1)

### Method The test sample and dilution water were mixed to produce each nominal loading rate and it was stirred by magnetic stirrer for about 48hours. Afterthefiltration, the filtrate was aerated for about one hour under about 242C in orderto recoverdecreased dissolved oxygen concentration by suction filtration, and the filtrate was used for the test solution. The test organisms were exposed to the test solution in order to confirm effect oftest item. The concentration oftest item in the test solutions were measured at the sametime.
(2)Result Nominal concentration (mglL) Left column: Cumulativemortality (%) Right column: Existenceofabnormal response abnormalities: 

# * no abnormalities: 

### N 3 hours 

### 24 hours 

### 48 hours 72 

### hours 96 hours 

### 1.2 0 , 

### N 

# · · 0 

# · N 

# · · 0 

# · N 

# · , 

# · 0 

# · N 

# · • 

# , 0 

# · N 

# · 3.6 0 , 

### N 

# · · 

# · 25 

# · N 

# · 

# · · 50 

### : 

# * 

# · 100 

# !



# - 100 

# i



# . 7.3 0 

# · N 

## 25 

# · N 

# · 

# · · 100 

# ·
- 

# · · 100 

# 1



# - , 100 : 

# - 

# · 12 0 

# · N 

# · · · 1001 

# - 100 

# · : 

# - 

# · 100 : 

# - 

# · 100 : 

# - , 

### Exposure system: semi-static(renewal at 48hours after) Numberoforganisms/volume oftest solution: 4 fish/About 2.5 L Aeration: conducted

- Alltest organisms died.

Nominal concentration Measured concentration (m ;/L) (mglL) At the start After

### 48 hours



### 1.2 0.30 n.d.
12 

### 1.7 1.0 (60) n.d.: <0.11 mgIL

2.2 Summary It 

### was confirmed that the measured concentration of test item decreased at 48 hours after. Therefore, the definitive study would beconducted as semi-staticregime (renewal at every 24hours) in consideration ofmaintenance ofthe concentration.



### 537 -2 Additional data 1­

## 94746

3. Operation ofthe definitive study

Based on the results of the preliminary study, the definitive study was conducted with five concentrations of the stock solution content 5.0, 3.1, 2.0, 1.2 and 0.8% (a geometric series with a factor of 1.6) and the control by semi-static regime (renewal at every 24 hours). Aeration was not conducted during the exposure. The preparation method ofthe test solution was the same as the preHminary study. The concentrations of test item in 

## the test solutions were measured at the start ofthe exposure, before and after the renewal and the end ofthe exposure.

538 -3 Additionaldata 1­

## 94746



# Additional data 2

Calculationresults ofconcentrations ofeach component



## 539

## 94746

1. Calculation results ofconcentrations ofeach component The test item was detected as two peaks in chromatogram. Calling Peak 1 and 2 in orderofelution, each peak corresponded to PE3MB and PEMB (Information supplied by the sponsor). Based on the resu1ts of HPLC analysis in definitive study in each exposure level, the concentrations of each component were calculated. Results of calculation are shown below. The concentrations of each component in each sample for HPLC analysis were determined on the basis of a comparison ofthe each peak area on the chromatogram of the sample solution with that of a standard solution. In addition. concentrationsofeach component in standard solution was calculated based on contentpercentage (PE3MB: 14.l%,PEMB: 81.7%).

Additional figure 2-1 Concentration ofPE3MB (Peak 1)in test solutions

Nominal 

## concentration (mgIL) Measured concentration (mglL) (Percentage ofmeasured concentration versus that at each preparation %) At the

start 24 hours 48 hours 72 hours At the end Geometric mean old new old new old new Control n.d. n.d. n.d. n.d. 

## n.d. n.d. n.d. n.d. 

# V

0.8 0.023 n.d.'3 

## 0.026 n.d:3 

## 0.044 0.029 (65) 0.031 0.025 (81) 0.022

1.2 n.d: 3 

## n.d. '3 

## 0.059 0.057 (96) 0.084 0.085 (101) 0.058 0.068 (118) 0.041

2.0 0.022 n.d. "3 

## 0.16 0.099 (63) 0.15 0.14 (92) 0.15 0.14*2 (89) 0.078

3.1 0.27 0.25 (93) 0.31 0.25 (80) 0.23 0.24*2 (109) 

# - - 0.26

5.0 0.30 0.23 (78) 0.44 0040 

## (91) 0.15 0.095 (65) 0.50 0047*2 

## (94) 0.28
old: before the renewal
new : after the renewal n.d. : <

## 0.018 mglL - : It

## showsno measurement because all test organisms died. *2 It

## indicates the measured value at the time that confirmed all test organisms dead. *3 In accordance with OECD Guidance Document No.23, halfthe determination limit ofthe test item (0.0090 mgIL = 

## 0.018 

## mglU2) was adopted for the calculation of geometric mean, because the test item was not detected and the detection Hmit (0.022 mgIL) was higher than halfthe determination limit ofthe test item.

540 -1 Additional data 2­

### 94746

Additional figure 2-2 Concentration ofPEMB (Peak 2) in test solutions Measured concentration (mglL) Nominal (Percentage ofmeasured concentration versus that at eachpreparation %) concentration (mg/L) At the 24hours start old
Control n.d. n.d.

0.8 0.13 0.043 (33)
1.2 n.d.'3 n.d.'3

2.0 n.d. '4 

### n.d.'3

3.1 0.41 0.34 (82)
5.0 0.22 0.16 (73) new
n.d.

0.14

0.19

0.44

0.42

0.47 48hours old new
n.d. n.d.
0.071 0.16 (52) 0.15 0.31 (81) 0.28 0.47 (64) 0.28 0.46 (67) 0.47 0.099 (101) 72 hours At the old new end

n.d. n.d. n.d.

0.082 0.15 0.094 (51) (62) 0.16 0.28 0.14 (52) (51) 0.22 0.45 0.31'2 (47) (69) 0.40'2



# - .

(86) 0.037 0.48 0.45"2

(38) (94) Geometric mean 

# /

0.10

0.083

0.13

0.38

0.22
old:beforethe renewal
new: afterthe renewal n.d. : < 

### 0.013 mg/L
- : It showsno measurement because all test organisms died. *2 

### Itindicates the measured value at the time that confinned all test organisms dead. *3 In accordance with OEeD Guidance Document No.23, halfthe determination limit of the test item (0.0065 mglL 

## = 0.013 

### mg/U2) was adopted for the calculation of geometric mean, because the test item was not detected and the detection limit (0.022 mglL) was higher than halfthe determination limit ofthe test item. *4 In accordance with OECD Guidance Document No.23, halfthe detennination limit ofthe test item (0.0065 mgIL 

## = 0.013 

### mg/U2) was adopted for the calculation of geometric mean, because the test item was detected butnot quantified.

541 

### -2Additional data 2­

### 94746



# Additional data 3

Qualitative analysis ofconvertedproducts

542

### 94746



### 1. Qualitative analysis ofconvertedproducts

The peaks which were likely converted pr09ucts except the test item were detected on the HPLC chromatogram of test solution in 

### the definitive study. Therefore, the sample 

## was qualitatively analyzed by liquid**chJomatograph~mass** 

### spectrometry**(LC~MS)** 

### and estimated structures.

1.1 Method ofanalysis Standard solution and the pretreated sample of 5.0 mgIL at the time that all test organisms died for HPLC analysis described in appendix 2

### were qualitatively analyzed by**LC~MS** 

### under the following conditions. Analytical conditions Instrument Liquid chromatograph**~** 

### mass spectrometer HPLC system SIDMADZU**LC~20A** 

### Series Mass spectrometer Thermo Electron TSQ Quantum Discovery Pump**(Post~column)** 

### HITACHI**~62oo** 

### Conditions ofliquid chromatograph Column L-column ODS (15 cm x 

### 2.1 mm I.D., Chemicals Evaluation and

Research Institute) Column temp. 40llC 

### Eluent A: Acetonitrile 

## I formic acid 

## (1000/1 v/v) B: 

### Water'! 

## Ifonnic acid 

## (1000/1 v/v)



### Gradient condition Time (min) A(%) B(%) 0.00 

### 50 50 20.00 100 

# o 40.00 

## 100 

# o Flowrate 0.2mllmin Injection volume 5pL Post-column Solution Acetonitrile 

## Iwater'} (1/1 v/v) "2 

### Flowratc 0.1 mUmin

*1 Tapwaterwas treated with ultrapure watersystem. *2 Containing 25 mmol/Lammonium acetate.

543 

## ·1 Additional dala 3·

### 94746

Conditions ofmass spectrometer Ionization mode Electrospray ionization (ESI) Detection ion Positive Measure mode Scan Scan range 

## (mlz) 150-600 Capillary temp. 3()()2C



### 1.2 Results Converted products, Peak 

### A, B, C, D, E and F detected on total ion current (TIC) chromatogram (Additional figure 3-4) were qualitatively analyzed by LC-MS and estimated as foUowing structures. Also, it wasrecognized that Peak 1 and 2 weretest item.

Peak No. Retention time on TIC chromatogram (min) Estimated structure and molecularformula Estimated molecular

weight Additional figure No.

A 

### 7.4

7.6 

# · · · · · · · · 

# · · · · · · · · · 

# · · · 

# · · · · 

# · · · · · · 

# · · · 

# · · · 

# · · 

# · Figure 3-5

Figure 3-13

Figure 3-6 Figure 3-14 

## B

,


# .

.

: 

# ·
:



### 1 

### 8.1 Figure 3-7 Figure 3-15

544 -2 Additionaldata :;­