NDA 21-372
`
`Page 61
`
`6. Ophthalmic Examination: No treatment-related changes were observed.
`
`7. Organ Weights: Females at the 3 and 10 mg/kg dose levels exhibited higher adrenal gland
`weights (10 and 13% increases) compared to controls. However there were no differences in
`organ/body weight ratios in these groups and no associated gross or microscopic abnormalities
`observed with respect to the adrenal gland in these animals.
`
`8. Gross/Microscopic Pathology: No gross or microscopic changes related to treatment with
`RS-25,259-197 were observed.
`
`In conclusion, the 3 mg/kg dose was the no effect dose for this study, and provides an adequate
`margin of safety for both the 0.3 ug/kg initial clinical dose and for the highest planned clinical
`dose, 120 ug/kg
`
`2.. Palonosetron Hvdrochloride: 28-Dav Subcutaneous Toxicity Studv in Juvenile Rats
`
`Report No: \ Report Number lO63/18-D6154
`
`Conducting laboratory and location: a _
`K
`
`Date Started: November 12, 1999
`
`Date of Study Report: July 2000
`Statutory Instrument 1999
`GLP compliance. Statements of compliance with t
`No 3106, The Good Laboratory Practice Regulations l999 and OECD Principles on Good
`Laboratory Practice (revised 1997, Issued January 1998) were included; however, there were no
`signatures.
`
`
`
`QA- Report: Yes (X) No 0
`
`Methods: In a 28-day subcutaneous toxicity. study, neonatal/juvenile rats received palonosetron
`at doses of 0, 5, 15, and 25 mg/kyday. The vehicle or drug solution was administered by the
`subcutaneous route for 31 days, starting at day 4 postpartum. In the clinical setting, the
`intravenous route of administration will be used; however, given the small size of animals in the
`study, the subcutaneous route was chosen. Rats (i.e., offspring) were killed and necropsied on
`day after the last treatment. Parental female rats received no treatment and were sacrificed
`following weaning of offspring.
`
`Dosing:
`species/strain: 30 time-mated female rats [Crl:CD(SD)lGSBR strain] were obtained from
`. to provide offspring for this study. The parental female rats
`were 8-10 weeks 01 and weighed at least 160 g at mating. Pregnant female rats were delivered
`by day 15 of gestation. These animals were allowed to deliver naturally and the day pups were
`first observed was designated as day 0 postpartum. On day 4 postpartum, litters were culled to 5
`
`

`

`NDA 21-372
`
`Page 62
`
`pups/sex/litter through random selection. Culled pups were discarded. Pups were examined on
`day 4 postpartum prior to allocation to the study. Offspring within each litter were allocated to
`the same treatment group. Litters were randomly allocated to treatment groups. The litter
`performance of parental female rats and the viability of offspring up to day 4 postpartum were
`poorer than expected with the result that less than required number of offspring were available
`for allocation. Accordingly, no satellite animals were allocated to the control group. The sex of
`each offSpring was recorded on days 1 and 4 postpartum prior to allocation to treatment groups.
`Sexes were confirmed at weaning (day 2] postpartum) and prior to necropsy (day 34
`postpartum).
`- #lsex/group or time point: In the main study groups, there were leuvenile
`rats/sex/group.
`- age: Rats were 4 days of age at the start oftreatrnent.
`- weight: On day 4 postpartum, mean body weights ranged from 7.3 to 8.3 g for male
`rats and 6.3 to 7.9 g for female rats.
`- satellite groups used for toxicokinetics or recovery: In toxicokinetic groups that
`received palonosetron at 5, 15, or 25 mg/kg/day, there were 15 or 16juvenile rats/sex/group.
`— dosage groups in administered units: 0, 5, 15, and 25 mg/kg/day
`- route, form, volume, and infusion rate: the subcutaneous route using a dose volume
`of5 ml/kg administered Vehicle or drug solution.
`Drug, lot#, radiolabel, and % purity: Palonosetron, batch number 30893—P105 (Purity 99.4%)
`Formulation/vehicle: The vehicle was sodium chloride and sodium phosphate adjusted to pH
`7.4 with sodium hydroxide or hydrochloric acid in Water for Irrigation.
`Observations and times:
`
`— Clinical signs: Parental animals and offspring were monitored daily for clinical signs
`oftoxicity. During the treatment period, pups were observed immediately and at l and 4 hr after
`dosing. Animals were observed for moribundity/mortality twice per day. Offspring that died or
`were sacrificed in a moribund condition during the treatment period were submitted to a
`macroscopic examination.
`- Body weights: Pup body weights were measured on day 1 postpartum and daily during
`the treatment period (days 4 to 34 postpartum).
`- Food consumption: Food consumption was not measured due to the age of the
`animals.
`
`'- Ophthalmoscopy: Not performed.
`- EKG: Not performed.
`- Hematology: Not performed.
`- Clinical chemistry: Not performed.
`- Urinalysis: Not performw.
`- Gross pathology: On the day following the last dose, surviving offspring were
`sacrificed and submitted to necropsy examination.
`- Organs Weighed: Organ weights were detennined for the adrenal glands, brain, heart,
`kidneys, liver, pituitary gland, prostate gland, spleen, testes + epididymides, and thyroids +
`parathyroids.
`- Histopathology: Gross lesions, principally at injection sites, from all animals and
`tissues from the control and high dose groups were embedded in paraffin wax BP, sectioned at 5
`pm, stained with hematoxylin and eosin, and examined by a pathologist. Deaths and moribund
`
`

`

`NDA 21—372
`
`Page 63
`'
`.
`sacrifices during the treatment period were considered unrelated to treatment, and
`histopathological examination was not extended to these animals. Tissues examined from the
`control and high dose groups were as follows: adrenal glands, brain, cecum, colon, duodenum,
`eyes, femur with bone marrow and articular surfaces, heart, ileum, jejunum, kidneys, liver, lungs
`with mainstem bronchi, mammary (females only), mandibular lymph nodes, mesenteric lymph
`nodes, esophagus, optic nerves, ovaries, pancreas, pituitary, prostate, salivary glands, sciatic
`nerves, skin, spinal cord (cervical, lumbar, and thoracic), spleen, stemum with bone marrow,
`stomach, testes and epididymides, thymus, thyroids and parathyroid, trachea, urinary bladder,
`and uterus.
`
`- Toxicokinetics: Blood for measurement of plasma levels ofpalonosetron and its
`metabolite, RS-l7825-007, was collected on days 1 and 28 (i.e., days 4 and 32 postpartum,
`respectively). Blood was collected from 4 or 5'rats/sex/group at 0.5, 1, 2, and 4 hr after dosing.
`Blood was obtained by decapitation on day 1 and from the orbital sinus on day 28. Samples were
`analyzed for palonosetron and RS—l7825—007 by.
`-_—_
`_
`’ Animals in toxicokinetic groups were discarded without examination following blood
`collection.
`
`- Other: Physical and functional development of rat pups was assessed. Eye opening
`was evaluated staning from day 13 postpartum. Evaluation of eye opening in the majority of
`pups was missed in error on day 16 postpartum. Air righting was evaluated on day 17
`postpartum. Pupillary reflex and auditory startle response were evaluated on day 21 postpartum.
`
`
`Results:
`
`- Clinical signs: For male and female rats at 15 and 25 mg/kg/day, dose-related
`incidences of hair coat thinning, hair loss, and sores were observed at injection sites. For rats at
`15 and 25 mg/kg/day, the percentage of animals with eyes open on day 15 postpartum was
`slightly reduced to 27 and 25%, respectively, as compared to 41% for the controls. By day 17
`postpartum, all rats in control and treatment groups had opened their eyes. The incidences of
`male and female rats at 15 and 25 mg/kg/day with an absent pupillary reflex on day 2]
`postpartum were increased as compared to controls. For male rats at 15 and 25 mg/kg/day, the
`pupillary reflex was absent for 30 and 20% of animals, respectively, as compared to a poor
`response for 10% ofcontrols. For female rats at 15 and 25 mg/kg/day, the pupillary reflex was
`absent for 10 and 30% of animals, respectively, as compared to a normal reflex for 100% of
`female controls. The sponsor reported background control ranges for offspring having no
`pupillary reflex of 0-42% for males and 046% for females. There were no treatment-related
`effects on air righting ability-on postpartum day 17 or auditory response on postpartum day 21.
`
`Pupillary reflex on postpartum day 2] for rats that received palonosetron by the subcutaneous route at doses of O, 5,
`
`
`
`(mean % of u-s showin_ a res-onse).- da15, and 25 m- '
`
`
`
`
`

`

`NDA 21-372
`
`Page 64
`
`- Mortality: There appeared to be no treatment-related mortality. Two female control
`rats (#123 and #124) were found dead, 1 on day 9 and l on day 6. Two female control rats (#121
`and #130) were sacrificed in a moribund condition, 1 on day 12 and 1 on day 27. One female
`control rat (#125) was missing and presumed cannibalized. One female rats at 5 mg/kg/day was
`sacrificed in a moribund condition on day 30, due to the poor condition ofits right eye following
`the orbital sinus bleed. Two male rats at 15 mg/kg/day (#62 and #64) were sacrificed in a
`moribund condition, 1 on day 7 and l on day 5. One male rat at 15 mg/kg/day (#61) was found
`dead on day 8. One male rat and one female rat at 15 mg/kg/day (#65 and #183) were found dead
`on day 6. One male rat and one female rat at 25 mg/kg/day (#91 and #212) died on day 30
`following anesthesia used for the orbital sinus bleed. The study protocol provided by the sponsor
`did not describe collection of blood samples from the orbital sinuses of rats in the main study on
`day 30. Further, the purpose of these blood samples was not described.
`
`- Body weights: Body weight gains were suppressed >10% for male treatment groups
`and female rats at 25 mg/kg/day. Body weights for male controls on days 4 and 34 postpartum
`were 7.3 and 121.0 g, respectively. Body weight gains for male rats at 5, 15, and 25 mg/kg/day
`were 88.6, 87.9, and 81.1% of the control, respectively. Body weights for female controls on
`days 4 and 34 postpartum were 7.2 and 108.6 g, respectively. Body weight gains for female rats
`at 5, 15, and 25 mg/kg/day were 94.3, 101.8, and 87.0% ofthe control, respectively.
`1.
`
`— Organ Weights: Changes in adjusted spleen and liver weights were observed for male
`and female rats at 15 and 25 mg/kg/day; however, there were no corresponding histopathological
`changes.
`
`- Gross pathology: For male and female rats that received palonosetron at 15 or 25
`mg/kg/day, thinning of the hair coat, hair loss, and sores were evident at injection sites (i.e., right
`and left shoulders and right and left hips). The incidence ofpelvic dilatation in the kidneys was
`increased for male and female rats at 25 mg/kg/day. The incidence ofprotruding eyes was
`increased for male and female treatment groups.
`
`
`
`9 8 7
`
`Gross pathological findings for rats that received palonosetron at subcutaneous doses of 0, 5, 15, or 25 mg/kg/day
`from days 4 to 34 .05 arturn.
`
`
`Female rats
`
`
`
`Skin + subcutis
`
`
`-sore
`
`
`-fur loss
`
`-red
`Lefi shoulder
`
`
`
`~fur loss
`
`
`-sore
`
`
`
`-red
`
`
`Right shoulder
`
`
`
`
`
`-fur loss
`-sore
`
`
`
`-red-__1_8_——-3-
`
`
`
`6
`
`9
`
`7
`
`9
`
`

`

`NDA 21-372
`Page 65
`
`-red
`
`-.rotrudin_
`
`
`
`
`-oelvic dilatation
`
`
`--ale
`
`
`
`
`
`.
`
`- Histopathology: Target organs (or tissues) oftoxicity were injection sites, optic
`nerves, and kidneys. For male and female rats at 15 and 25 mg/kg/day, histopathological changes
`at injection sites included mineralization, hemorrhage, acanthosis, dermatitis, and cellulitis.
`Cellulitis was characterized by loss of the squamous cell layer with inflammatory cell infiltration
`ofthe underlying epidermis and demu's with scab formation and infiltration into the underlying
`muscle. Additionally, mineralization of the hair follicles and muscle fibers was observed. For
`male and female rats at 25 mg/kyday, the incidence of neuropathy (slight to minimal, focal) in
`the optic nerves was increased. For male and female rats at 25 mg/kg/day, the incidence of pelvic
`dilatation in the kidneys was increased. The incidences of focal nephropathy and papillitis were
`increased for female rats at 25 mg/kgday.
`Histopathological findings for rats that received palonosetron at subcutaneous doses of O, 5, 15,
`or 25 mg/kgday from days 4 to 34 postpartum.
`
`
`O_,(ION 3
`
`Male rats '
`
`Female rats
`
`Skin + subcutis
`N =
`
`10
`
`2
`
`9
`
`5
`
`2
`
`—_—__—_
`
`—hemorrhage
`dermatitis
`-cellulitis
`
`Left shoulder
`
`
`N =
`
`
`-mineralization
`-hemorrhage
`
`
`-acanthosis
`-dermatitis
`
`-cellulitis
`
`Right shoulder
`
`
`N =
`
`
`-mineralization
`
`
`4
`0
`'0
`0
`5
`2
`0
`O
`-hemorrhage
`
`
`9
`6
`0
`0
`7
`4
`0
`0
`-dermatitis
`‘
`9
`7
`0
`0
`7
`4
`0
`0
`-cellulitis
`
`___-___—
`
`
`l
`2
`l
`
`9
`
`5
`6
`l
`9
`7
`
`9
`4
`
`0
`O
`0
`
`0
`
`0
`O
`0
`' 0
`0
`
`0
`0
`
`I
`\
`
`0
`
`0
`O
`0
`0
`0
`
`0
`0
`
`6
`
`0
`2
`l
`2
`S
`
`6
`0
`
`l
`2
`1
`
`7
`
`0
`0
`0
`5
`4
`
`8
`0
`
`9
`
`6
`2
`1
`8
`8
`
`9
`7
`
`0
`0
`0
`
`0
`
`0
`0
`0
`0
`0
`
`0
`0
`
`.
`
`0
`2
`0
`
`0
`
`0
`0
`0
`0
`0
`
`0
`O
`
`
`
`
`
`
`
`
`
`

`

`NDA 21-372
`Page 66
`N =
`-mineralization
`
`-hemorrhage
`—acanthosis
`-dennatitis
`-cellulitis
`
`-mineralization
`
`—hemorrhage
`-acanthosis
`—dermatitis
`-cel|ulitis
`
`OOOOOO
`OOOOOO
`
`
`
`-pelvic dilatation
`—focal nephropathy
`- . a . illitis
`
`- Toxicokinetics: For male and female rats, plasma AUC values for palonosetron on
`days I and 28 increased with elevating dose, although, observed increases were greater than
`proportional to dose. Plasma AUC values for palonosetron were similar on days 1 and 28. There
`appeared to be no gender-related differences in plasma AUC values for palonosetron. Due to
`limited sample volumes available on day l oftreatment, AUC values for palonosetron in the low
`and high dose group males were not determined and toxicokinetic analysis for RS-17825-OO7
`was not performed at any dose level. The sponsor reported that low concentrations of the
`metabolite, RS-l 7825-007, were detected in the low and intermediate dose groups on day 28 to
`the extent that toxicokinetic analysis could be conducted.
`
`
`
`
`
`
`
`
`Plasma pharmacokinetic parameters for palonosetron in rats that received palonosetron by the subcutaneous route at
`doses of 5, 15, or 25 ma/kelday.
`Dose
` Ba 28
`
`
`
`M g/kg
`Tm, hr
`AUCQHM, ng'hr/ml
`Cm,
`Tm, hr
`AUCOZHHD ng'hr/ml
`Cw,
`
`n- ml
`n_ ml
`Z
`3
`
`~11
`
`
`_F -_—II- —_F —---
`
`IiKl!
`1
`0.
`.5
`.5
`.
`—2.
`.
`38.63 -0
`4707
`646 76
`75210
`776.50
`
`1357.68
`
`558.71
`
`1393.40
`
`655.68
`
`700.90
`
`
`
`-—-——--————-m
`NC = Not Calculated
`
`Key Study Findings: In a 28-day subcutaneous toxicity study, neonatal/juvenile rats received
`palonosetron at doses of 0, 5, 15, and 25 mg/kg/day. The vehicle or drug solution was
`administered by the. subcutaneous route for 3] days, starting at day 4 postpartum. 1n the clinical
`setting, the intravenous route of administration will be used; however, given the small size of
`animals in the present study, the subcutaneous route was chosen. Rats (i.e., offspring) were killed
`and necropsied on day after the last treatment. Parental female rats received no treatment and
`were sacrificed following weaning of offspring. The no effect dose was 5 mg/kg/day. There was
`
`

`

`NDA 21-372
`
`Page 67
`no treatment-related mortality. The incidences ofmale and female rats at 15 and 25 mg/kgday
`with an absent pupillary reflex on day 21 postpartum were increased as compared to controls.
`Eye opening was slightly delayed for rats at 15 and 25 mg/kg/day. Target organs (or tissues) of
`toxicity were injection sites, optic nerves, and kidneys. For male and female rats at 15 and 25
`mg/kg/day, histopathological changes at injection sites included mineralization, hemorrhage,
`acanthosis, dermatitis, and cellulitis. For male and female rats at 25 mg/ngday, the incidence of
`neuropathy in the optic nerves was increased. For male and female rats at 25 mg/kg/day, the
`incidence ofpelvic dilatation in the kidneys was increased. The incidences of focal nephropathy
`and papillitis were increased for female rats at 25 mg/kg/day. Hematology and clinical chemistry
`‘ parameters were not provided, although, it appears that blood was collected on day 30 of
`treatment.
`
`3. Subcutaneous 28-Dav Toxicitv Studv in Juvenile Rats with Palonosetron Hvdrochloride:
`
`(Study #PALO-OZ—OS; PV10001)
`
`Testing Laboratogy:
`
`\
`
`Dates of Stan and Completion of the studv: April 10, 2002 & December 5, 2002
`
`GLP # QAU Reguirements: Statement of compliance to "GLP regulations, 1999, "1999 #
`3106 was attached.
`
`Species and Strain: Cr12CDRBR VAF/Plus SD IGS BR VAF/PLUS strain neonate rats
`
`
`Batch #: TA/2002/163 (Batch #21000565)
`
`Methods: Sixty mated females were allowed to litter and 136 pups/sex were divided in to 4
`main groups (lZ/sex/group) and 4 satellite groups (22/sex/group). These were administered
`subcutaneous injection (between the shoulder blades) 0, 5, 15 or 25 mg/kg/day palonosetron (5
`mng) for 28 days (post-partum day 4 to day 34/35). The following table of sponsor (vol 58.1,
`pp 18 of 422) shows group size, doses and identification # of animals.
`
`

`

`NDA 21-372
`
`Page 68
`
`'
`
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`Sponsor conducted this repeat study on the request ofthe Agency (letter January 31, 01 l) as the
`previous study (t — report #1063/18-D6154) was not complete and it lack the
`measurements of hematology and clinical chemistry parameters. The doses and the methods
`used in the present study were the same as used in the previous 28-day study in juvenile rats
`( ~ ~ report #1063/18-D6154).
`In brief, the study included the assessment ofth'e body
`weights changes, the developmental parameters of ear/eye opening (from day 3 to 6 for ear and
`from day 1 1 to 16 for eye opening), righting reflux (day 5), air righting reflex (three trials on day
`22), startle response (on day 15), pupillary light reflex and opthalmoscopy (on day 33 day of age)
`were also observed. The Approximately 0.5 ml of blood was collected from 4 satellite group
`from 3 pups/sex/time point on day l, 4 and 35/36 of dosing (i.e., postpartum day 4) at 0.5, l, 2
`and 4 hr after dosing for the estimation ofpalonosetron and, its metabolite RS 17825-007. At
`necropsy (day 35/36 of animal age), 0.5 ml the blood from the non-fasted main study groups for
`hematology and blood chemistry parameters. The tissues and organs from main study groups
`were the same as described the previous study. The microscopic examination was done on all
`tissues from control and high dose treatment groups ofmain study animals and, eyes and optic
`nerves from all the animals of all groups of the main study and, all tissues of animals from main
`study group dying or killed during the study and, all gross lesions from the main study groups
`animals.
`
`Results:
`
`a. Observed Effects: Two males (#67 and 72) and 1 female (#265) Thickening and scabies of
`the skin at the injection sites were seen in animals of 25 mg/kg/day treatment group. No other
`changes in the physical condition of the animals were observed.
`The percent animals with ears Open (day 5), startle response (day 15), eye opening (day 16) were
`100% among all the groups. The average day for opening the eyes were 14.9, 15.2, 15.0 and
`15.2, respectively in control and 3 treatment groups. The righting reflex on day 5 of observation,
`was 8.3% less in animals of 25 mg/kg/day treatment group than control group animals. The
`percent animals with eye open on day 14 were 22.2, 8.3, 16.7 and 16.7 in control and 3 treatment
`groups animals. The development parameters were similar in all study groups.
`
`

`

`NDA 21-372
`
`Page 69
`
`b. Mortality: Three animals were found dead during the study. These were 2 males in control
`group and 1 female of satellite group of5 mg/kg/day treatment group. Two males of control
`group and 1 female on5 mg/kg/day treatment group (satellite group) were sacrificed because of
`humane reasons.
`
`c. Bodv Weight/Food Consumption Changes: The mean body weight gain ofthe males and
`females included in treatment groups and control group were not statistically different during the
`preweaning period. The mean initial (day l) and final body weights (on postpartum day 20) of
`neonates of the control main study group during the pre-weaning period were males 7.7 g and
`46.4 g and, females 7.1 and 44.2 g, respectively. During post weaning period (postpartum day
`21 to 35), a retardation of 10.7 and 5.8% in the body weight gain of male and female animals
`included in 25 mg/kg/day treatment group was seen. On post weaning day 35, the bodyweights
`ofanimals ofmain study control and 3 treatment groups were 145.5, 143.8, 148.1 and 132.7 g
`for males and, 133.4, 138.5, 133.8 and 126.0 g for females, respectively.
`
`TABLE
`
`Table for Mean Wei ht () of rats durin the study
`IIIIIIIIIIIIIllflflflllflflflflflflillflflfliflfllllflfiflififll
`Pre-Weaning
`Postnatal Day 3 M 7.7
`F
`
`Postnatal Day 20 M
`F
`
`Day 35
`
`Post-Weaning
`Day 21
`
`.
`
`M
`
`d. Hematoloolv/Coagulation/Bone Marrow Changes: A slight decrease was seen in RBC
`count, hemoglobin and percent packed cell volume in treated animals. A dose related increase in
`total WBC counts in treated males (10.65, 11.02, 12.8 and 13.8X103ul) and females (9.87, 12.45,
`11.68 and 12.98 X103/ul) was also seen.
`
`e. Blood Chemistrvarinalvsis Changes: Alkaline phosphatase (ALP) levels were decreased
`(p<0.001) by 6.6, 18.0 and 24.5% in males and, 6.0, 24.7 and 21.7 in females of 5, 15 and 25
`mg/kg/day treatment groups. The albumin to globulins ratio was decreased in a dose related
`manner in males and females indicating the drug effect on liver. The serum low albumin and
`ALP values may be due to the inflammatory reactions caused by the compound.
`
`f. thsical Examination and Ophthalmic Test Changes: The abnormality of eye lens opacity
`and posterior synechia was seen in 1 out of 12 male pups and, the delay in development of
`unilateral lens suture line was obServed in 1 out of 12 female pups of 25 mg/kg/day treatment
`group. No optic nerve neuropathy was seen in the pups during the study.
`
`

`

`NDA 21-372
`
`Page 70
`
`g. Organ Weight Changes: The heart (p<0.01), liver and kidney (0.05) absolute weights were
`decreased1n males of 25 mg/kg/day treatment group. The relative weight of these organs to
`body weight was not affected The thymus and lung organ absolute weights were decreased
`(p<0.05 to 0001) among females of 25 mg/kg/day treatment group.
`
`Toxicokinetics: The subcutaneously administered compound was seen to attain the peak plasma
`levels within 0.5 to 1 hr ofits administration on day 4 and 30. The plasma peak levels on a
`steady state (AUCo. 11:) and the time for peak plasma concentration of the compound were
`similar1n males andfemales on day 4 and 35 as shown1n sponsor table 3 (scanned below).
`
`Table 3: Results [tom the non-compartments! analysis of plasma concentration-mne-
`dara (palanoselron).
`‘
`
`Day Group
`
`Dose
`(mg/kg)
`
`('1)
`
`C...
`109m“)
`
`AUCu
`(ng/mI-h)
`
`AUG...
`(nB/mLh)
`
`1,
`(M)
`
`n
`
`Iv,
`(h)
`
` 0.50
`
`1.00
`1.00
`
`355.04
`1415.35
`0.50
`050 147125
`0.50
`252.54
`
`334.37
`156425
`2333.35
`24520
`
`1.266 4'
`450.27 0.545
`1593.01
`1.078 0.643
`4
`2393.65 0.995 0.697
`4
`250.99
`1.590 0.410
`3
`
`359.59
`1539.32
`
`110425
`2220.54
`
`112.35 1.153 0.601 3|
`23141.70 0.896 0.774
`a
`
`Female
`
`4
`
`2
`3
`4
`
`5
`15
`25
`
`0.50
`0.50
`0.50
`
`414.55
`931.11
`1680.31
`
`557.41
`1420.54
`2559.61
`
`50529 0.675 1.025
`1437.40 0.336 0.329
`3125.39 0.340
`1.033
`
`l
`=
`L
`
`35
`
`286.77 2293 0.302
`276.35
`249.01
`0.50
`5
`2
`1139.45 1241
`0.559
`1126.97
`945.93
`050
`15
`3
`1525.09
`1.134, 0.535
`1797.22
`1544.02
`1.00
`25
`4
`‘ Corresponds to the number ol points used in the hall-lite delenninetton.
`
`4
`4
`4
`
`2
`3
`3
`
`Table 4: Results from the non-compartments! analysis olplnsma concentration-time
`dam (Rs-1711250177).
`
`Sax
`
`1
`
`
`Day Group' 03.59" T._.. C...
`AUG...
`AUG“
`A.
`in
`n -
`11119419)
`01)
`(no/m1)
`(nelmI-h)
`("91"”)
`(m)
`(h)
`
`l
`
`02:17
`
`35.73
`
`0.564 0.302
`
`0.888
`
`1.038
`
`/
`
`
`wNNQUm
`humuum
`UUWUUU
`AWFJAUN
`
`
`25
`
`5
`
`1.00 33.53
`
`1.00
`
`17.93
`
`59:44
`
`3320
`
`‘ Corresponds to the number at points used in “he hall-tile deterrninatlon.
`
`The peak plasma concentration of the metabolite (RS-17825-1 87, with the following structure)
`was seen within 1 to 2 hr of the administration of the compound.
`
`

`

`NDA 21-372
`
`Page 7]
`
`RS- 178254137
`
`
`
`The toxicokinetic parameters of the metabolite were shown in sponsor table 4 (as scanned
`above).
`
`
`h. Gross Patholo v Findin s: An abnormal skin color was seen in 2 and 4 males and, 1 and 4
`females of 15 and 25 mgi’kgday treatment groups. The incidences of scales formation were 1
`and 7 among males and, 2 and 6 females of 15 and 25 mg/kg/day treatment groups. Abnormal
`size ovaries and uterus were seen in 2 females of 25 mg/kg/day treatment group.
`
`i. Histopatholog'ical Changes: Renal pelvic distension was present in 0 of 12, l of 1, 1 of2,
`and l of 12 males and, 0 of 12, 1 of 1,
`1 of2, and 3 of 12 females belonging to 0, 5, 15 and 25
`mg/kg/day treatment groups. The incidences ofrenal pelvis distension in animals died during
`the study were I of] and 1 out of2 males of5 and 15 mg/kg/day treatment groups. One of2
`females of 15 mg/kg/day treatment group died during the study. Extramedullary haemopoiesis in
`spleen of marked intensity was observed in 1 male and 5 females out of 12/sex of25 mg/kg/day
`treatment group. Slight to moderate intensity dermal acanthosis at the site of injection was
`observed in 4 of4 and 12 of 12 males and, 2 of2 and 6 of8 females belonging to 15 and 25
`mg/kg/day treatment groups. Dermal ulceration was seen in 50, 100 and 75% males and, 0, 50
`and 62.5% females of 5, 15 and 25 mg/kg/day treatment groups. Myofibre degeneration/
`pannicular necrosis of subcutaneous tissue from moderate to marked intensity was observed in
`50% and 83.3% males and, 50 and 100% females belonging to 15 and 25 mg/kg/day treatment
`groups. Optic nerve neuropathy was not seen in any of the study animals.
`
`APPEARS THIS WAY .
`0N ORlGlNAL
`
`

`

`NDA 21-372
`
`Page 72
`
`Histopatliological findings that were considered to be related to treatment were seen at the
`subcutaneous injecnon sites as tabulated below :
`I——'_—'—_—— _ —.
`
`E_
` l No Examined
`
`i Males
`
`
`
`l panrucular
`
`
`lluemorrliage/
`
`! congestion
`'
`-
`
`0
`'
`-
`.\l_\-'olibic
`.lpnnnnr:when]
`
`
`
`-necrosis
`Denna);
`
`
`Subcutaneous
`necrosis
`
`
`
` Dystrophic
`5 mineralisation
`
`
`
`In summary, subcutaneously administered palonosetron caused renal pelvic distension,
`extramedullary haemopoiesis of marked intensity in spleen and dermal reactions including
`myofibnl degeneration/necrosis of moderate to marked intensity in animals of 15 and 25
`mg/kg/day treatment groups. Eye lens opacity and structural abnormality of posterior synechia
`was seen in l of 12 male pups and, delay in development of lens suture line was seen in l of 12
`female pups of 25 mg/kg/day treatment group. No incidences of optic nerve degeneration were
`seen. A dose of 5 mg/kyday was a ‘highest tolerable dose’ and kidney, spleen and site of
`injection were the. target organs of toxicity.
`
`26-Week Chronic Intravenous Toxicitv Studv in Rats with 4 Week of Recovefl
`4.
`Period: (Study # 1063/1-D6154; PALO 99—08)
`
`Testing Laboratorv:
`
`
`
`

`

`
`
`NDA 21-372
`
`Page 73
`GLP and QAU Reguirements: The study was conducted in compliance with GLP and QAU
`audits requirements (the certificate was not signed)
`
`Dates of Start and Completion of the Study: May 17, 1999 and January 12, 2000 (The date of
`completion of pathology not given)
`
`" with mean body weight of277.3
`.——-o
`Animals: Crl:CD(SD)IGSBR strain rats ‘
`to 289.7 g (males) and 197.8 to 205.8 g (females) were used in the study.
`
`Methods: Two hundred and sixty animals (l30/sex) were randomly divided in to 4 groups i.e.,
`30, 20, 20 and 30 animals/sex in 0, 2, 7 and 14/10 mg/kg/day palonosetron (B. #1308911/30893-
`P105; vol. 1 ml/kg) treatment groups, respectively. Ten animals of control and high dose
`treatment groups were reserved for the recovery period of 4 weeks to determine the reversibility
`oftreatment related effects.
`In addition, 10 rats/sex/group consisted of satellite treatment groups.
`The dose selection was based on a preliminary dose escalating toxicity study performed in 2
`groups of rats (5/sex/group) and these animals were given a dose of7 or 10 mg/kg/day for 7
`days. These doses were escalated to 14 and 20 mg/kg/day, respectively in these two groups on
`day 8. Uncoordinated movements were seen after 14 mg/kg/day dose from day 11. Based on
`this observation 3 dose of 10 mg/kg/day was selected as the dose for the present study.
`
`Group
`Number
`
`Description/Dose
`(mg/kg/day)- Main study
`Male
`Female
`
`Animals/group
`Satellite Study
`Male
`Female
`
`1
`2
`3
`4
`
`Control
`Low
`lnterrnediate
`High
`
`0
`2
`7
`10
`
`20+ 10#
`20+10
`20
`20
`20
`20
`20+10# 20+10#
`
`-
`10
`10
`10
`
`-
`10
`10
`10
`
`-the dose levels are expressed in term of free base. A conversion factor (1.123) was applied to convert from the base
`to salt; # 10 animals/sex from the control and high dose groups maintained treatment free for 4 weeks following 26
`weeks of treatment.
`
`All animals were observed for the changes in the clinical signs, mortality, food consumption, and
`body weight changes. The ophthalmoscopic examination was done in animals of control and
`high dose treatment groups on week 25. Blood samples were collected on day 1, and on 15' day
`of week 5, 13 and 26. The blood samples from 3 animals/sex/satellite group of 10 mg/kg/day
`treatment group at 10, 30 min and 1, 2, 6 and 24 hr after dosing for toxicokinetic experiment.
`Blood samples for hematology and blood chemistry tests were drawn from 10 animals/main
`study treatment groups. Urine samples were collected from other 10 animals /sex/group (not
`used for blood parameters estimations) at week 12 and 25 and week 30 (afier recovery period).
`All animals of the study (including satellite group) were necropsied and examined visually. The
`organs and tissues of each of the animals were separated, cleaned, weighed and/or preserved for
`microscopic examination. The organsweighed were adrenals, brain, kidneys, liver, pituitary,
`testes/ovaries, prostate, spleen, thyroid/parathyroid and uterus. The organs fixed in formalin,
`methanol and Davidson’s stain before microscopic examination were adrenal, bone marrow
`
`

`

`NDA 21-372
`
`Page 74
`
`(femur), eye/optic nerve, zymbal glands, muscle Quad, skin and mammary glands, sciatic nerve,
`seminal vesicle, femur marrow and artificial surface, ileum, cecum, colon, rectum, uterus,
`prostate, urinary bladder, stemum+Marrow, testes+epididymedis, liverXZ, salivary glands,
`mesenteric/mandibular lymph nodes, spleen, thyroid/parathyroid, pancreas, duodenum, jejunum,
`spinal cord (cervical, thoracic, lumber), lungs, bronchi, kidneys, nasal turbinate, nasopharynx,
`esophagus, stomach, aorta, larynx, pituitary, ovaries, brain, heartX3, thymus, trachea and gross
`lesions.
`
`Results:
`
`a. Observed Effects: Four out of 10 animals included in 94 mg/kyday treatment group had
`convulsions on first day of treatment. Uncoordinated movements and/or reduced activity were
`seen in these animals immediately after dosing. Therefore the dose 14 mg/kg/day was reduced to
`10 mg/kg/day from study day 2.
`
`b. Monalig: Eight animals ofthe study died and these were 1 female belonging to control and 7
`animals of 14/10 mg/kg/day treatment group. The animal number, week of death and cements
`on each of the animals died, are shown in the following table (table taken from sponsor
`submission vol 1:], pp 050):
`
`Group Number Animal no/sex
`1
`125F
`4
`76m
`77M
`173F
`199F
`223M(S)
`229M(S)
`259F(S)
`
`(S) = satellite group animal
`
`Mortality Table
`Week of Death
`Comments
`30
`Died during bleed procedure
`14
`Found Dead post dose
`14
`Found dead post dose
`14
`Found dead post dosing.
`26
`Died dun'ng bleed procedure
`22
`Found Dead post dosing
`12
`Found dead prior to dosing
`23
`Found Dead post dosing
`
`Three animals (# 173, 77 and 76 died approximately 4 hr post dosing with no clinical signs. Animal # 223 showed convulsions
`before death. The other animals had no signs before deaths.
`
`C. Body Weight/Food Consumption/Water consumption: The animals included in 2, 7 and
`14/10 mg/kg/day treatment groups gained similar body weights. On week 26, the mean final
`weights were 517.6, 517.4, 522.1 and 502.1 g for males and 298.3, 301.4, 301.4 and 301.1 g for
`females included in 0, 2, 7 and 14/10 mg/kg/day treatment groups, respectively. On week 26,