`
`Reviewer: Lilliam A. Rosario PhD. NDA No.21-344
`
`Study title:
`
`1C] 182,780: One Month Intramuscular Toxicity Study in Rats.
`
`Key study findings:
`0
`Females exhibit histological changes in the ovaries (absent or reduced corpora lutea, multiple
`follicular cysts, luteal cysts, hemorrhagic Graaftan follicles and hemorrhagic corpora lutea),
`uterus (atrophy), cervix (atrophy) and vagina (atrophy).
`0 There are no differences in the incidence or severity of the histological changes observed in
`rats administered ICI 182,780 alone or in combination with sulphone. These changes appear
`to be related to the anti-estrogenic activity of the compound.
`
`Study no:
`TAR/2972
`Volume #, and page #: N_000\2001-03-28\pharmtox\tox\Dose\ TAR/2972
`
`Conducting laboratory and location: AstraZeneca UK Limited Safety Assessment Alderley
`Alderley Park Macclesfield Cheshire SK10 4TG England
`Date of study initiation: April 21, 1999
`GLP compliance: Yes
`)
`QA report: yes (x) no (
`Drug, lot #, radiolabel, and % purity: Batch #. P/ 1465/28, P/ 1465/26, P/ 1465/29, 999% w/w
`Formulation/vehicle:
`
`
`Ingredients
`
`1031 182780
`ICI |82780 sulphone
`Poloxamer 407 USNF
`Ethanol 96% 85'
`Water for injectitm Ph Eur
`Propylmeglycol
`Formulation hatch reference
`
`Strength
`placelm
`% w’v
`
`:
`1.0
`100
`3.0
`w 100 %
`
`”465/28
`
`Strength
`2 % wa'v
`% w‘v
`
`2.0
`-
`I ’0
`'0'0
`o
`8-0
`‘°‘°°/°
`szsxzs
`
`Strength
`1.90 "A. Mi;
`% w‘v
`
`1.96
`0.04
`1.0
`10.0
`8.0
`moon/a
`P/l 465/29
`
`Methods: This study compares the toxicity of lCI 182,780 2% SA (short acting) formulation and
`a ICI 182,780 1.96% (with 0.04% sulphone) SA formulation administered
`intramuscularly to the rat once daily for 28 days. Note: The sulphone derivative is a
`degradation product of ICI 182,780 formulations and is a minor metabolite in the rat after
`N administration. The results of this study are intended to support the final specification
`for sulphone content in ICI 182,780 parenteral formulations. The dose level selected for
`the study of5 mg/kg/day (30 mg/m2) x 28 days = 140 mg/kg (840 mg/mz) is ~ 30x-(5x)
`the proposed human dose of 250 mg/30 days (4.17 mg/kg or 154.3 mg/mz).
`Dosing:
`Species/strain:
`#/sex/group or time point (main study):
`Satellite groups used for toxicokinetics:
`Age:
`Weight:
`Doses in administered units:
`
`Alpk: APfSD -Wistar derived rats
`10/sex/group
`3/sex/group
`37-41 days old
`127-217 g
`0, 5 mg/kg/d, and 5 mg/kg/d ICI
`18270 + 0.4 mg/ml sulphone. Daily
`x 28 days
`
`42
`
`

`

`NDA No.21-344
`Reviewer: Lilliam A. Rosario PhD.
`
`Route and volume:
`
`Observations and times:
`
`Clinical signs:
`Body weights:
`
`Food consumption:
`Ophthalmoscopy:
`Hematology:
`
`Clinical chemistry:
`Urinalysis:
`Gross pathology:
`Organs weighed:
`
`Histopathology:
`Toxicokinetics:
`
`Results:
`
`Mortality:
`
`Clinical signs:
`
`Body weights:
`
`Food consumption:
`Ophthalmoscopy:
`Hematology:
`
`IM; 0. 025 ml/100 g
`
`Twice daily
`Day -7, pre- study, first day of dosing and then weekly for the
`remainder of the study.
`Daily
`Pre-study and week 4
`At scheduled necropsy. Blood samples for coagulation analyses
`were taken from designated 5/animals/group at scheduled
`necropsy (day 29)
`At scheduled necropsy (day 29)
`At scheduled necropsy (day 29)
`Day 29
`Adrenal glands, brain, heart, kidneys, liver, lungs, ovaries,
`pituitary gland, prostate gland, spleen, testes (including
`epididymides), thymus and uterus.
`Day 29
`Pre-dose and 1, 3, 6, 12, and 24 hours on day 28. The AUC0_24
`of ICI 182,780 and the AUCo..2 of the circulating metabolites
`(ZM208,917 sulphone and ZM366,472 17— ketone) for the male
`and female rats were compared within and between each dose
`group.
`
`One 0‘ animal dosed 5 mg kg ICI 182,780 with Sulphone, from
`the pharmacokinetic sub- group, died (day 5) as the result of an
`accident. The Sponsor did not indicate the nature of the
`accident. Necropsy findings included minimal focal acute
`myositis and minimal multifocal adjacent tissue acute
`inflammatory cell infiltration at the injection site. Also, mild
`unilateral hydronephrosis and minimal focal cortical tubular
`basophilia
`
`Clinical observations included swelling of the hind limbs with
`associated transient limping in several animals in all groups
`including controls from day 3 to 24. The Sponsor considers this
`effect a consequence of exposure to the excipients and not ICI
`182,780.
`
`Males from both groups dosed with ICI 182,780 showed an
`approximate 10% body weight gain reduction when compared
`to control group.
`i
`All females gained weight at a similar rate throughout the
`dosing period.
`Unremarkable (UR)
`UR
`
`Values represent percent change from control.
`
`43
`
`

`

`
`
`Reviewer: Lilliam A. Rosario PhD. NDA No.21-344
`
`Hemoglobin (g/dl)
`RBC (x 10‘2/1)
`Reticulocytes (%)
`Hematocrit (l/l)
`MCV (n)
`MCH (pg)
`Platelets (109/1)
`WBC (109/1)
`
`T9'
`T14.
`T35‘
`T19
`Group Il-drug + sulphone; Group Ill-drug alone
`‘P<0.05
`
`.
`
`Values represent ercent Chan e fiom control.
`
`
`
`
`
`
`Glucose (mmoll)
`Urea (mmol/l)
`'
`
`T. calcium (mmol/l)
`ALP (TU/l)
`Triglycerides
`mmon
`
`IIIA
`
`Control
`
`11
`
`Ill
`
`llA
`ilo
`
`Control
`14.6
`6.3
`30
`l
`
`3.28
`344
`1.05
`
`
`Group ll-drug + sulphone; Group Ill—drug alone
`‘P<0.05
`
`Clinical chemistry:
`
`Urinalysis:
`Organ weights:
`
`UR
`At necropsy, there was a decrease in median body weight of
`male rats in Group Il(11%) and in Group III (16%) given ICI
`182,780. Thus, some absolute organ weights for male rats in
`these groups showed significant differences from the control
`values. The Sponsor considers these changes to be of no
`toxicological importance.
`Median relative uterine weight was decreased by 79% for
`females in Group H and 1]] given ICI 182,780.
`
`Gross pathology:
`
`
`
`
`Group ll-drug + sulphone; Group Ill-drug alone
`
`
`
`
`Histopathology:
`
`
`
`
`——-I_-E_
`
`
`__——
`
`
`Ovaries
`
`
`
`No corporea lutea
`Reduced corporea lutea
`Follicular Cysts
`
`
`Luteal Cysts
`
`
`Hemorrhagic Corporea Lutea
`
`
`Hemorrha_'c Graafian follicles
`Uterus: Severe Atrophy
`
`Vagina: Epithelial Severe Atrophy
`Estrus
`
`
`
`
`
`_
`0/10
`5/10
`
`10/ l 0
`0/10
`
`
`10/10
`9/9
`
`0/9
`
`
`
`

`

`
`Reviewer: Lilliam A. Rosario PhD.
`NDA No.21-344
`
`Injection site-acute myositis
`
`Adjacent tissue acute inflammatory cell
`infiltration
`
`Sciatic nerve-adjacent acue inflammatory
`cell infiltration
`
`5/10
`
`5/10
`
`5/10
`
`
`
`
`
`
`
`
`_—-_-E_
`
`Injection site-acute myositis
`Adjacent tissue acute inflammatory cell
`infiltration
`
`3/10
`1/10
`
`4/10
`4/10
`
`5/10
`5/10
`
`cell infiltration
`
`Group ll-drug + sulphone; Group Ill-drug alone
`
`
`
`
`
`
`
`Toxicokinetics:
`
`ZM 36 ,472'
`Group
`Grou
`11‘
`ll]
`
`Group
`11]
`
`.
`-IEIlm-IEI 3-0
`.
`
`‘with 0.04% sulphone; ” 1C1 182.780 was measured 0-24 and ZM 208.917 and ZM 366,472
`were measured 0-12
`
`" ZM208,917 is a sulphone metabolite
`B ZM366,472 is a 17-ketone metabolite only demonstrated in females.
`
`0 There was no difference in exposure to ICI 182,780 between the sexes or between the groups
`given lCI 182,780 alone or in combination with sulphone.
`0 Absorption appeared faster in animals given ICI 182,780 and sulphone compared to
`administration of the drug alone. That is, on day 28, Cmax for ICI 182,780 occurred at 1 h
`post—dose (tmax) for Group H (drug + sulphone) compared to 3 h post-dose for Group 1]] (drug
`alone).
`0 Exposure to ZM208,9] 7 (sulphone metabolite) was slightly greater in females compared to
`males (Group II and HI : 62% and 59%, respectively).
`0 Exposure to ZM208,917 (sulphone metabolite) was approximately 70% greater in animals
`given ICI 182,780 in combination with sulphone than in animals given ICI 182,780 alone.
`0 There was a significant difference between the sexes in the AUsz of ZM366,472 (ketone
`metabolite) with male rats showing no systemic exposure. However, there was no difference
`between the AUCO- .2 of ZM366,472 of Groups H and III females.
`
`Summary of individual study findings:
`Study TAR/2972 compares the toxicity of the ICI 182,780 formulation (2% ICI 182,780-SA) and
`a ICI 182,780 formulation (SA) spiked with sulphone (1.96% ICI 182,780 with 0.04% sulphone),
`when administered intramuscularly to the rat once daily for 28 days. The sulphone derivative is
`the main degradation product of ICI 182,780 formulations and is a minor metabolite in the rat.
`
`Pharrnacokinetic monitoring demonstrated no difference in exposure to ICI 182,780 between the
`sexes or between the groups given ICl 182,780 alone or in combination with sulphone. Exposure
`to ZM208,917 (sulphone metabolite) was greater in females compared to males and ~ 70%
`
`45
`
`

`

`
`
`Reviewer: Lilliam A. Rosario PhD. NDA No.21-344
`
`greater in animals given ICI 182,780 in combination with sulphone than in animals given ICI
`182,780 alone (the increase in exposure probably due to administered ZM208,917). Exposure to
`ZM366,472 (ketone matebolite) was observed only in females.
`
`Changes seen in animals given ICI 182,780 (alone or in combination with sulphone) were related
`to the anti-estrogenic activity of the compound. The changes seen included a 10% reduction in
`body weight gain in males and histological changes in the ovaries (absent or reduced corpora
`lutea, multiple follicular cysts, luteal cysts, hemorrhagic Graafian follicles and hemorrhagic
`corpora lutea), uterus (atrophy), cervix (atrophy) and vagina (atrophy). There were no differences
`in the incidence or severity of histological changes between the two groups.
`
`Study title:
`RATS.
`
`ICI 182,780 : SIX MONTH INTRAMUSCULAR TOXICITY STUDY IN
`'
`
`Key study findings:
`0 Atrophy of the female reproductive tract; a specific constellation of ovarian alterations with
`increased late stage and cystic Graafian follicles, loss of mature corpora lutea and reduced
`vacuolation of the interstitial cells.
`
`0 A loss of spermatozoa from the seminiferous tubules with an accompanying dilatation,
`seminiferous tubular atrophy with some associated degenerative changes in the epididymides.
`0 Conversion of specific tissues in females to the morphology normally seen in males including
`mammary gland structure, splenic hemosiderosis, pituitary gonadotroph vacuolation and
`reduced hair loss and an earlier appearance of adrenal cortical congestion with hemocyst
`formation.
`
`TPR/2042.
`Study no:
`Volume #, and page #: N_000\2001-03-28\pharrntox\tox\Dose\ TAR/2972\TPR/2042.
`Conducting laboratory and location: ICI Pharmaceuticals Safety of Medicines Department
`Alderley Park Macclesfield Cheshire England
`Date of study initiation: April 2, 1992
`GLP compliance: Yes
`QA report: yes (x) no ( )
`Drug, lot #, and % purity: ICI 182,780; ADM 44010/89; 98.8%
`Formulation/vehicle: Sustained release LA formulation
`
`Ingredients
`1C1 182,780
`Ethanol 96% v/v
`
`Benzyl Alcohol
`Benzyl Benzoate
`Castor Oil
`Batch number
`
`Strength w/v%
`5.0
`10.0
`
`Placebo w/v"/.
`0
`10.0
`
`10.0
`15.0
`To 100
`PH/6731/41
`
`10.0
`15.0
`To 100
`PH/6731/40
`
`ADM 48026/90
`
`Analytical Number
`
`ADM 48027/90
`
`Methods:
`
`Dosing:
`Species/strain:
`
`CR] :(WDBR Wistar rats
`
`46
`
`

`

`Reviewer Lilliam A Rosario Ph D
`.
`.
`,
`.
`
`.
`
`NDANo.2l-344
`
`#/sex/group or time point (main study):
`Satellite groups used for recovery:
`
`20/sex/group
`lO/sex/vehicle control or 10
`
`mg/rat/30 days ICI 182,780
`retained for a 19/20 week
`
`withdrawal period after the 6th
`dose.
`
`Satellite groups used for PK:
`Age:
`Weight:
`Doses:
`
`5/sex/time point
`36—43 days old
`110-199 g
`0 (saline and vehicle), 15 mg/kg/3O
`days, 10 mg/rat/30 days or 10
`mg/rat/15 days.
`0 All main test animals received 7 doses at 30 day intervals of ICI 182,780 or control
`formulation, except Group V which received 13 doses at 15 day intervals. Animals
`were observed for at least 7 days prior to necropsy.
`0 Animals in withdrawal groups received 6 doses of ICI 182,780 or control formulation,
`at 30 day intervals and were observed for 19 or 20 weeks after cessation of
`dosing/before necropsy.
`0 On day 61, 5 male rats from Group III were erroneously dosed with the vehicle
`control. These animals were allowed to recover and then correctly dosed on day 64,
`giving inter-dose periods of 33 days before and 27 days after this dose. This error is
`not considered to have compromised the study.
`Route, form, volume, and infusion rate:
`
`IM using a plastic syringe and a
`sterile needle 25G x 16 mm; 0.2
`ml/rat for Groups I, 11, IV and V
`and 0.03 ml/l 00 g body weight for
`Group III.
`
`Dose Levels
`1C! 162, 780
`
`0 lg/rat/JO days
`vehicle control
`
`0 I‘lrlz/BO days
`saline control
`
`Hain test
`
`Vithdr-vcl
`
`20 H, 20 F
`
`30 H, 30 F
`
`20 H, 20 F 10 H, 10 F
`
`15 Ig/kg/BO days
`
`20 H, 20 F
`
`0 H,
`
`O F
`
`10 .g/nt/ao days
`
`30 n. so 1:
`
`10 lg/xnt/IS days
`
`IO H, )0 F
`
`0 H,
`
`0 T
`
`Note: The dose of 15 mg/kg/30 d was chosen as a dose which could be administered once a month for 6 months within the
`constraint of the maximum permitted dose volume (0.1 ml/sitetn 2 sites). The 10 mg/rat dose was set to maximize exposure of
`the rats with doses at the limit volume every 30 or 15 days
`
`Observations and times:
`
`Clinical signs:
`Body weights:
`
`Food consumption:
`
`Twice daily
`Day -7 and -2 pre-study, on the first day of dosing, then weekly
`to week 13 and then monthly for the remainder of the study.
`Recorded 7 days pre—study, then weekly to week 13, and
`monthly for the remainder of the study.
`
`47
`
`

`

`NDA No.2 l -344
`Reviewer: Lilliam A. Rosario PhD.
`
`Ophthalmoscopy:
`
`Hematology:
`
`Clinical chemistry:
`
`Urinalysis:
`
`Gross pathology:
`Organs weighed:
`
`Histopathology:
`Toxicokinetics:
`
`Results:
`
`Mortality:
`
`Clinical signs:
`
`Body weights:
`
`Pre-study and all group I, l], and V animals in the main study
`were also examined during weeks 14 and 26 of the study
`Weeks 13 and 24 of dosing and from the withdrawal animals
`on week 40.
`
`Weeks 13 and 24 of dosing and from the withdrawal animals
`on week 40.
`
`Weeks 13 and 24 of dosing and from the withdrawal animals
`on week 40.
`
`Main group: week 27-29; withdrawal: week 46-47
`adrenal glands, brain, heart, kidneys, liver, ovaries, pituitary
`gland, prostate gland, spleen, testes (including epididymides)
`and uterus.
`
`Main group: week 27-29; withdrawal: week 46-47
`Samples were taken 3, 7 and 24 h after the first dose and on
`days 4, 7, 11, 16, 21, 25 and 30. In addition, samples were
`taken 3 and 24 hours after the second dose from rats in Group
`V. Samples were also taken after the sixth (Groups HI and IV)
`and eleventh/twelfth (Group V) dose at the same time intervals
`given for the first and second dose above.
`
`There were two premature sacrifices during the study, M
`400826 (vehicle control) and F 400775 (10 mg/rat/30 days) due
`to broken teeth and body weight loss.
`Bruising around the injection site was observed in one o'
`(vehicle control).
`
`
`Cumulative Bod
`Wt. Gains ;
`
`
` - Females
`
`Week
`Vehicle
`15
`10
`10
`Vehicle
`15
`10
`10
`- rug/kg!
`mg/rat/
`mg/rat/ - m g!
`
`mg/rat/
`
`days
`30da s
`lSda s
`dn 5
`30 da 5
`lSda s
`__-__—_mm
`-_—__—_I_IIE-
`"WE-IE!!-
`_—M
`—-_-_-E_-—
`_m-—IMI__-_
`MEN-___-
`_Im-m-m—___
`mum—__—
`“__EEIIEEI_—__
`Il-——__—-_—
`”Em—__-
`-—_-_____
`mum-___—
`__m——__—_
`
`“MIMI!“ 417
`412
`* p<0.05, "‘p<0.01,‘”‘“‘p<0.00l
`
`
`
`
`
`
`
`
`
`
`
`
`mg/rat/
`
`
`
`
`
`
`
`
`
`
`48
`
`

`

`
`
`Reviewer: Lilliam A. Rosario PhD. NDA No.21-344
`
`0 Female rats given 15 mg/kg/30 days showed impaired growth throughout the study with
`a cumulative gain over the 25 week measurement period of 81% of control value.
`Females given 10 mg/rat, at 30 or 15 day intervals, showed impaired growth over the
`study (cumulative gains of 87 and 92% of controls).
`0 Male rats given ICI 182,780 showed a slight impairment of growth up to 4 weeks after
`the first dose compared to controls.
`0 During the withdrawal phase male rats previously given 10 mg/rat/30 days gained less
`weight than the vehicle controls (gain between weeks 25—41 ~ 85% of control) while
`females from this group showed similar growth to vehicle controls.
`
`Food consumption:
`0 Male rats given ICI 182,780 ate similar amounts of food to controls during the dosing
`period. On weeks 2 and 3, male rats given 10 mg/rat (at both 15 and 30 day intervals)
`had significantly lower (4-6%) group mean food consumption values compared to
`controls.
`
`0 Female rats given ICI 182,780 ate less food (9-15%) than controls during the dosing
`period. For rats given 15 mg/kg/30 days and 10 mg/rat/30 days, this effect began in
`week one and continued throughout the dosing period. Females given 10 mg/rat/15 days
`also had lower (9%) group mean food consumption from week 2 to week 8, after which
`time it was comparable to the control animals.
`0 During the withdrawal phase the food consumption of animals previously given 10
`mg/rat/30 days (males and females) was lower than that of the control animals.
`Ophthalmosc0py:
`UR
`
`10m.m115d
`10mm/30a
`1—-m-m=m-
`— “—-_ F “u“
`Hemoglobin (g/dl)
`13
`15.46
`14.25
`14.91.
`l5.06“‘
`14.55m
`15.24”.
`14.73”
`15.27m
`24
`15.13
`14.24
`14.09m
`14.97"
`14.25m
`15.82‘”
`I4.58'
`15"
`13
`8.94
`7.32
`13.57.
`23.40m
`8.41‘”
`24
`9.14
`8.04
`8.58"
`8.]79
`8.16
`13
`0.4480
`0.411
`0.4296"
`24
`0.4695
`0.429
`0.4360'“
`
`13.51m
`
`8.64‘"
`
`8.63”
`
`0.4307
`
`0.4100m
`
`0.4534“
`
`0.4220m
`
`0.4300
`
`
`
`
`
`
`
`Hematolo 3 :
`
`RBC (x10 l1)
`
`Hemalocn't (V1)
`
`Recov
`
`
`
`
`
`
`
`8
`
`
`
`
`
`
`
`
`
`Hemoglobin (g/dl)
`
`RBC(x10 ll)
`
`
` Hematocril (V1)
`
`
`
`- p<0.05, "p<0.01,"'p<0.001
`
`“u“
`15.472
`14.102
`14.55m
`15.240m
`15.300
`14.383
`14.25m
`15.269
`8.902
`7.83
`8.48"
`8.48“
`9.146
`8.229
`8.51‘”
`8.486
`.4070
`.44 94
`0.4150‘”
`.4320“
`.4629
`.4343
`0.4160‘”
`.4629
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`13
`24
`13
`24
`13
`24
`
`0 Males given ICI 182,780 showed lower hemoglobin (4-7%), hematocn't (4-11%), and
`RBC (4-7%) values compared to vehicle controls. Decreases in RBC counts may be
`suggestive of hemolysis unrelated to vehicle.
`0 Female rats given lCI 182,780 showed higher hemoglobin (5-1 1%) concentrations
`compared to controls at both time-points in all dose groups. At week 13, this was
`associated with an increase (~8%) in red blood cell count however, at week 24, this
`increase was only observed in rats given 10 mg/rat/30 days (17%).
`
`
`
`
`
`
`
`49
`
`

`

`
`
`Reviewer: Lilliam A. Rosario PhD. NDA No.21-344
`
`0 The same pattern of changes was observed in male and female rats in the recovery group.
`However, it is noteworthy that the values for 6‘s in the main test (10 mg/rat/30 days) and
`withdrawal group (10 mg/rat/30 days) are identical.
`
`Clinical chemist
`
`15m k 30d
`
`roman/30d
`
`10m rat/15d
`
`Total protein (g/l)
`
`Albumin (g/l)
`
`Alkaline
`hos hatase (lU/l
`Calcium (mmol/l)
`
`
`
`-_—__—_-I--l-
`13
`72
`70.10
`69.60
`6360’”
`68.5'
`65.30‘“
`71
`65.6"
`24
`76.34
`75.90
`73.50‘
`69‘“
`73.80
`69.80‘”
`74.7
`68.4‘“&
`13
`35.8
`33.90
`32.6‘
`32’”
`32.8
`33.1‘”
`33.7
`32.7‘”
`24
`35.24
`38.1
`34.1
`3390‘”
`34.2
`34.6‘”
`35
`33.5‘”&
`13
`245.9
`194.6
`236.4
`234.8‘
`285.7'
`247.4"
`250.2
`266.9‘“
`24
`248.6
`166.6
`233.6
`210.9‘
`265.6
`200.3
`231.2
`230.3“&
`13
`2.644
`2.669
`2.676
`2.548"
`2.643
`2.584’
`2.558‘
`2.508'”
`24
`2.773
`2.775
`2.727
`2.624‘”
`2.725
`2.596‘”
`2.767
`2.60"‘&
`13
`2.34
`1.97
`2.07
`1.95
`1.80“
`1.91
`1.66‘”
`1.97
`24
`2.908
`2.43
`2.38‘
`2.03
`2.13"
`1.92‘
`2.140”
`2.05
`
`
`
`
`
`
`
`
`
`0.986
`0.905”
`
`1.262
`1.542
`
`
`
`0.837'
`0.990‘
`
`
`
`
`
`1.476
`1.510
`
`0.813‘
`0.819“&
`
`
`
`
`
`
`
`
`
`Cholesterol
`(mmol/l
`1.537
`1.218
`1.624
`13
`Triglycerides
`1.355
`1.480
`1.651
`24
`(mm/1)
`p<0.05, “p<0.01,”‘p<0.001, &=continues into the withdrawal period
`0 Female rats given ICI 182,780 had lower plasma total protein (16-10%) and albumin
`concentrations (14-12%) and higher alkaline phosphatase activities (TZl-38%) compared to
`controls. These differences were still apparent at the end of the withdrawal period suggesting
`that full recovery had not occurred. According to the Sponsor, the lower plasma total protein
`and albumin concentrations may be attributed to the anti-estrogenic effect of the drug in the
`liver since protein synthesis is partially regulated by estrogens. The higher alkaline
`phosphatase activities may be a result of the removal of estrogen suppression allowing
`elevation to levels associated with males.
`
`0 Lower (5-7%) total calcium values than controls were observed in all females given ICI
`182,780 and these small differences are probably related to the reduction in plasma
`albumin/total protein, which serve as carrier proteins for the calcium.
`0 Males given 1C1 182,780 exhibited lower (1 8-29%) cholesterol concentrations than controls.
`At week 24, the values were similar to those observed in control females.
`
`0 Females given ICI 182,780 also showed lower (19-45%) triglyceride values than controls in
`weeks 13 and 24 and at the end of the withdrawal period this difference was still apparent
`between control and rats previously given 10 mg/rat/30 days.
`
`Urinal sis:
`
`
`
`-- Vehicle
`15 m
`. 30 d
`_-III-———-Z--I--_
`77.4‘(sz)
`37.5
`43.2( 43%)
`93.6'(T3x)
`70.3(T2x)
`102.4‘(T3x)
`43.2(T43%)
`
`
`15.3
`36.2
`94.9” T6x
`60 T4):
`38.4 T3):
`32
`52.2. T3x
`
`
`
`
`50
`
`

`

`
`
`Reviewer: Lilliam A. Rosario PhD. NDA No.21-344
`
`Spleen (l)
`
`0.95“
`
`C
`0.56”
`
`0.963"
`
`0.629‘
`
`0.976‘
`
`0.625“
`
`1.068
`
`.745
`
`1.028
`
`.685
`
`Liver (l)
`
`22.82
`
`12.39
`
`21.63
`
`22.16
`
`21.21
`
`10.83“
`.‘
`
`23.80
`
`12.39
`
`20.48
`
`12.59
`
`
`
`
`
` Utems (i)
`
`0.1205
`0.086’
`0.148‘
`I’
`.l&
`.‘
`
`Adrenals
`005
`.087
`.053
`.087
`054
`.095
`.052
`.097
`.0721
`.0782
`.0620
`.1 121'
`
`Pituitary (i)
`0.013
`0.015
`0.0113
`0.0112
`0.012
`0.0106
`0.0112
`0.0103
`.0136
`.0174
`.0129
`.0148
`
`
`
`fit.
`ti.
`3’
`t..
`ti!
`’0‘
`‘p<0.05, "p<0.0| ,"“p<0.001 , &= trend continues into the recovery period
`
`
`
`
`
`0.539
`
`53 9a
`
`
`
`Gross pathology:
`
`Compared to controls, changes included a reduction in size of
`the female reproductive tract, changes in the size, constituency
`or color of the ovaries and testes, and a lower incidence of hair
`loss in the females that approximated to that observed in the
`males. The majority of these changes were also observed at
`termination of the withdrawal stud .
`
`_ Males n=20
`Females n=20
`vehicle
`15
`10
`10
`15
`10
`10
`
`mg/kg/
`mg/rat/
`mg/rat/
`mg/kg/
`myrat/
`mg/rat/
`30 min
`30 da
`15 da
`30 min
`30 da
`15 da
`
`___—___nn
`___—_—-_-_m-—
`——————m--_-_
`___—_“__—
`___-___“—
`”nu-___-
`”___-___—
`mun-"___—
`mum—__—
`
`
`
`
`
`
`
`
`
`___-_u—_-—
`
`
`
`a8 IOG.
`
`
`—----
`
`mg/rat/
`30d
`
`mg/rat/
`30d
`
`comosition
`
`___-n
`___—l-
`___" 3
`_——n-E-
`___-‘-
`Emu—m
`-—_-__—
`___--
`___-__—
`___—_—
`
`
`F
`2/20
`
`20/20
`
`10m_ rat/15d
`
`5/20
`
`I!
`
`18/20
`
`5]
`
`

`

`
`
`Reviewer: Lilliam A. Rosario PhD. NDA No.21—344
`
`11/20
`
`6/20
`
`2/20
`
`0/20
`
`0/14
`
`0/20
`
`6/20
`
`4/20
`
`- - -
`0/20 - 20/20 - 20/20 - 20/20
`
`
`
`0/20
`
`mild
`
`9/20
`
`
`
`1/20
`
`mild
`
`4/20
`
`0/1 1
`
`4/20
`
`mild
`
`
`
`epididymides
`ductal eith. microc stic
`reduced/
`loss of s-ermatozoa
`
`male like mammary
`
`1m
`ovaryreduced
`
`interstitial cell
`vacuolation
`
`follicles
`
`co oralutea
`
`pituitary pars distalis
`
`vacuolation/foaminess
`
`of cells
`
`testes seminiferous
`tubular atrophy
`
`loss of spermatozoa
`with tubular dilation
`
`uterus atrophy
`
`vagina atrophy
`
`injection site cysts
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`15/20
`
`0/19
`
`0/20
`
`0/20
`
`1/20
`
`4/20
`
`13/20
`
`5/20
`
`- - -
`
`5/20 (3
`severe, 2
`min-mild
`6/20
`(2 mod, 4
`min—mild
`
`9/20
`(8 severe,
`1 mod
`6/20
`(3 mod, 3
`min-mild
`
`5/20
`(4 severe,
`1 mod
`l0/20
`(5 mod, 5
`min-mild
`
`0/20 - 20/20 - 20/20 - 20/20
`oo.
`38
`3388
`20/20
`20/20
`mod
`
`20/20
`
`0/20
`
`3/20 mild
`
`2/20 min
`5/20 mild
`
`5/20 mild
`1 1/20
`mod
`
`
`
`3/20 min
`‘
`2/20 min
`myocyte necrosis
`
`
`inflammation
`14/20
`18/20
`14/20
`
`
`
`'
`'
`mild
`'
`'
`mild
`mild
`
`
`
`2/20 mod
`1/20 mod
`
`
`
`Histohatholo in Withdrawal Grou.
`
`
`
`
`
`
`Recove
`adrenal cortical
`con_estion/hemoc sts
`reduction}
`loss of s ermatozoa
`cervix atroh
`
`mammary gland
`_ladular vauolation
`ovary reduced
`interstitial cell
`vacuolation
`
`ovary reduced/loss of
`mature co nora lutea
`inc. later sta_e follicles
`pituitary pars distalis
`vacuolation
`seminiferous tubular
`mineralization
`seminiferous tubular
`atrophy
`
`loss of s-ennatozoa
`
`1/10
`
`1/9
`
`
`
`6/10
`
`3/9
`
`0/10
`
`0/10
`
`0/10
`O/IO
`
`
`
`
`
`10m rat/30 d
`F
`7/ l 0
`
`0/10
`
`3/10
`
`0/10
`
`6/10 mild
`6/10
`
`9/10
`
`5/10
`
`3/l0
`2/10
`
`3/10
`
`5/10
`(4 severe,
`1 mild
`2/10
`
`
`
`
`
`
`52
`
`

`

`
`Reviewer: Lilliam A. Rosario PhD.
`NDA No.21-344
`
`0/10
`
`0/10
`2/10
`
`10/10
`mild
`8/10 mild
`8/10
`
`6/10
`
`utuerus atrophy
`
`va_inal atroh
`
`site
`
`
`
`O
`
`with some dilation of
`
`seminiferous tubules
`
`
`
`
`
`
`
`
`9/10 mild
`6/10mi1d
`5/10mi1d
`4/10mi1d
`2/10mod
`2/20mod
`l/lOmod
`
`In animals of all groups, changes at the injection sites included moderate cyst formation
`around the deposited material with mild inflammation. Rats given 10 mg/rat/ 15 days, by
`subjective assessment, appeared to show a greater number of identical cysts as a result of
`more frequent intramuscular administration. Following the withdrawal period, cysts were still
`present at the injection sites but the cysts appeared slightly more fibrous with evidence of
`resolution of the inflammation.
`
`0
`
`In all females given ICI 182,780, there was a diffuse atrophy of the uterus, cervix, and vagina
`with a loss of normal cyclical estrous activity. In the ovaries, there was an increase in the
`number of later stage Graafian follicles (some cystic and others showed an apparent
`anovulatory leuteinization). A reduction in the number, or loss of mature corpora lutea and a
`loss of the normal ovarian interstitial cell vacuolation also occurred. These changes persisted
`to termination of the withdrawal study, although there was a decrease in the number of
`animals with increased numbers of later stage/cystic follicles.
`0 A proportion of males from all groups given ICI 182,780 showed a unilateral or bilateral
`reduction in numbers of mature spermatozoa within the seminiferous tubules with an
`accompanying dilatation. Also, a significant increase in incidence of multifocal or diffuse
`atrophy of the seminiferous tubules occurred. Microcystic degeneration of the epithelium of
`epididymal ducts was common in these rats. Changes in the testes and epididymides also
`persisted to termination of the withdrawal study.
`0 Conversion of specific tissues in females to the morphology seen in males (loss of sexual
`dimorphism) including mammary gland structure, splenic hemosiderosis, pituitary
`gonadotrOph vacuolation and reduced hair loss, and an earlier appearance of adrenal cortical
`congestion with hemocyst formation was observed in animals receiving ICI 182,780.
`0 At the end of the withdrawal period, drug—treated females showed a higher incidence of
`microvacuolation of the mammary glandular epithelium even though general mammary gland
`morphology had returned to the normal state, Also, the increased incidence of pituitary
`gonadotroph vacuolation persisted in the withdrawal animals. Although there were no
`histological differences between the control and closed withdrawal group adrenal glands, the
`increased mean adrenal weight for the high dose group may indicate a persistent increased
`severity of adrenal cortical congesion with hemocyst. The lowering incidence of splenic
`haemosiderin deposition (approximating males) did not appear to persist to termination of the
`withdrawal study.
`
`53
`
`

`

`
`
`Reviewer: Lilliam A. Rosario PhD. NDA No.21—344
`
`Toxicokinetics:
`
`The following table shows the ~ actual dose (mg/kg) administered to Groups IV (10
`mg/rat/30days) and Groua V 10 m rat/15 da 5
`
`
`Female
`
`
`w2 A09V
`
`dose
`
`dose
`
`dose
`
`10
`
`10
`
`10
`
`10
`
`
`
`wwmnwwmm
`
`N
`_N\lN
`
`dose
`
`lSda s
`
`-_
`
`_- \lM
`
`_‘ \J\l
`U! \J
`....
`>— 1..)
`
`
`
`
`30:11! 5 30da'515davslSda s 30da 5 30112 5
`lSda s
`m— m —-
`\l
`
`4 —m
`_ as
`256
`MNv.0
`33#0
`
`
`
`0
`
`0
`
`It is noteworthy that as body weight increases (~2-3 fold) with time, the actual dose
`administered decreases in both male and female rats.
`
`Since females have lower (~30%) body weights compared to male rats, the actual dose
`administered to female rats is thus greater than males.
`
`Serum Concentration 11 ;
`mm man-.-
`-I1E_———-_—
`l-—— 15.501152
`_ 49.91513
`
`
`
`
`
`
`
`l-_—
`16.4:2.69
`I--——— _—_
`I-__
`_——_ l3.4:t2.37 I_——
`n-——
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`Group
`
`m I
`
`momma/sod)
`I-l-I-
`
`
`
`m—nnnnnnnnnnnnnn II
`n
`372 88.3 208
`56.3 60.2 94.5 125194 70.2 129 94.8 132
`105
`
`ME“!!! . .3 H“ m
`
`AUC‘(ng‘d/ml)
`326
`719
`638 1235-1352-1317-1931
`1684
`—n 5.4 III-1m 5-63 9-86 man-I-
`-
`
`# 1.13 = 3 h afler dosing; 1.30 = 7 h after dosing
`‘AUC in the dosing interval: 0-30 d for groups Ill and IV; 0.16 d for group V.
`0 There was a significant difference between the sexes in serum ICI 182,780
`concentrations, Cm“, and AUC. The Sponsor attributes these differences to the lower
`body weight of female rats compared to male rats, given a fixed dose (10 myrat).
`
`
`m
`
`
`
`
`
`
`
`
`54
`
`

`

`
`
`Reviewer: Lilliam A. Rosario PhD. NDA No.21-344
`
`0
`
`When body weight was taken into account, however, there was still a trend towards
`higher exposure in female rats.
`Tmax showed considerable variability occurring between 3 h (day 1.13) and 11 days
`after dosing. Following the peak, the serum ICI 182,780 concentrations declined
`slowly following the first and sixth doses for rats given 15 mg/kg/30 days and 10
`mg/rat/3O days and the first and twelfth doses for rats given 10 mg/rat/ 15 days. The
`decline was slower following the higher dose and more frequent dosing interval
`(Groups IV and V respectively)
`0 The group mean AUC in the dosing interval 0 — 30 days showed a proportional
`increase with dose.
`
`0 Following the 6th (or 12th) dose, AUC values were higher in both males and females
`compared to the first dose for Groups 1]] and V. This trend was, however, reversed in
`Group IV, where AUC values following the sixth dose were lower than those
`following the first dose. The reason for this difference is unclear.
`
`Summary of individual study findings:
`Impairment of growth in females at all doses accompanied by lower food intake. Slightly higher
`red blood cell counts and mean cell hemoglobin concentrations in females at all doses
`(reversible). Lower plasma total protein, albumin, triglycerides and calcium, and higher alkaline
`phosphatase activity in females at all doses.
`
`Histologically in rats at all doses: changes at the injection sites included moderate cyst formation
`around the deposited material with mild inflammation still present by the end of the recovery.
`Also, atrophy of the female reproductive tract; a constellation of ovarian alterations with
`increased late stage and cystic Graafian follicles, loss of mature corpora lutea and reduced
`vacuolation of the interstitial cells. Conversion of specific tissues in females to the morphology
`normally seen in males including mammary gland structure, splenic hemosiderosis, pituitary
`gonadotroph vacuolation and reduced hair loss and an earlier appearance of adrenal cortical
`congestion with hemocyst formation. A loss of spermatozoa from the seminiferous tubules with
`an accompanying dilatation, seminiferous tubular atrophy with some associated degenerative
`changes in the epididymides. Decreased weights of spleens, pituitary glands and, in females only,
`livers, which were unaccompanied by overt toxicological changes. Atrophy of the cervix, uterus,
`vagina, as well as, ovarian changes showed evidence of reversibility but not complete recovery at
`the end of the withdrawal period. Similarly, reduction/loss of spermatozoa and seminiferous
`tubular mineralization and atrophy were still present in male rats.
`
`55
`
`

`

`
`
` Reviewer: Lilliam A. Rosario PhD. NDA No.21-344
`
`Study title:
`
`ICI 182,780 : ONE MONTH TOXICITY STUDY IN DOGS.
`
`Key study findings:
`0 Repeated daily dosing over 4 weeks resulted in non-proportional increases in systemic
`exposure and an ~3-fold accumulation in AUC0-24. The majority of the accumulation took
`place during the first two weeks of closing.
`0 Drug-induced histopathological changes comprised testicular Leydig cell hyperplasia and
`multiple ovarian Graafian follicles. These effects are likely a result of the antiestrogenic
`action of ICI 182,780.
`
`0 Chronic myositis was observed at injection sites.
`
`TAD/583
`Study no:
`Volume #, and page #: N_OOO\2001-O3-28\pharmtox\tox\Dose\ TAR/2972\TAD/5 83
`Conducting laboratory and location: ICI Pharmaceuticals Safety of Medicines Department
`Alderley Park Macclesfield Cheshire England
`Date of study initiation: August 22, 1989
`GLP compliance: Yes
`)
`QA report: yes (x) no (
`Drug, lot #, and % purity: ICI 182,780 analytical reference number ADM 44026/88; 99.5%
`Formulation/vehicle:
`
`."'\
`
`Ingredients
`
`lCl l82,780
`Placebo
`m %w/v m_2%w/v
`
`———
`—-a--ru
`_-m--nu
`—mm
`
`49033/89
`
`—-—reference number
`44026/88
`
`Reference Number
`
`6124/l44
`
`6124/l45
`
`Reference Number
`
`49028/89
`
`Methods:
`
`Dosing:
`Species/strain:
`#/sex/group or time point (main study):
`Satellite groups used for recovery:
`Age:
`Weight:
`Doses:
`
`Alderley Park Beagle dogs
`3/sex group
`3/sex/group for 0 and 2.5 mg/kg/d
`49—61 weeks old
`11.5-21.2 kg
`0, l, 2.5, and 4 mg/kg/d; daily x 4
`weeks
`
`Route, form, volume:
`
`1M and SC once daily
`
`56
`
`

`

`
`
`Reviewer: Lilliam A. Rosario Ph.D. NDA No.21-344
`
`
`
`o0125
`
`0. 025
`
`0.0375
`
`0.1
`2 sites
`0.15
`2 sites
`
`Total
`volume
`ml/k d
`
`0.125
`
`0.2
`
`Observations and times:
`
`Clinical signs:
`Body weights:
`Food consumption:
`Ophthalmoscopy:
`
`EKG:
`
`Hematology:
`
`Clinical chemistry:
`Urinalysis:
`Gross pathology:
`Organs weighed:
`
`Histopathology:
`Toxicokinetics:
`
`Other:
`
`Results:
`
`Mortality:
`Clinical signs:
`
`The main test animals received 28- 31 daily doses. Withdrawal
`animals received 28 doses and were then left undosed for 42 — 43
`
`days prior to necropsy.
`
`Twice daily
`Weekly
`Daily
`Pre-study period and on day 28 of dosing. Wi