NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
`The Sponsor's dose selection is based on high multiples of the antiritl i-Pri 1iioh firleP in hnnmanc
`(b) (4)
`adjusted for body surface area (BSA). The proposed high dose of
`(b) (4)
`than the predicted maximum human dose of 9 mg/day (5.55 mg/m2 based on a 60-
`kg bodyweight), on a body surface area. basis. It is noted that systemic exposure of plecanatide
`in the animal species tested has been found to be very low after oral administration, which is
`suggestive of limited oral bioavailability. In humans, plecanatide was not detectable in plasma.
`(10 ng/ml limit of quantification) in any of the clinical studies. Thus, in this particular situation,
`where the human AUC can not be measured due to the absence of detectable plasma drug levels,
`the rodent to human AUC ratio should not be used for dose selection. The Sponsor provided no
`(b) (4)
`specific explanation for the proposed middle and low dose oflMling/kg/day,
`respectively.
`
`Dose selection in males cannot be based on an MTD, since no MTD was established due to the
`absence of adverse effects in the dose-ranging study.
`
`Executive CAC Recommendations and Conclusions:
`
`1. The Committee did not concur with the proposed doses for the 2-year carcinogenicity study
`in rats. The Committee recommended doses of 0, 10, 30, and 100 mg/kg/day for female rats,
`by oral gavage, based on reduced body weight gain in females at 300 mg/kg/day. The
`Committee recommended doses of 0, 10, 30, and 100 mg/kg/day for male rats based on an
`anticipated large rat to human margin of local drug concentration in the intestinal tract, and
`the expectation that the drug concentration in rats will achieve a maximum pharmacological
`effect. A rat to human local intestinal plecanatide concentration ratio was estimated
`according to the intestinal fluid volumes in rats and humans. The following table shows an
`estimation of the local exposure ratio based on the predicted maximum clinical dose of 9
`mg/day and an oral dose of 100 mg/kg/day in rats.
`
`Estimated Intestinal Plecanatide Concentration Based on GI Tract Volume
`
`Fluid
`volume-
`Fasting
`(ml)
`163
`3.2
`
`Fluid
`volume-
`Fed
`(ml)
`751
`7.8
`
`Estimated
`Drug Cone
`Fasting
`(mg/m1)
`0.055
`6.25
`
`Estimated
`Drug Cone
`Fed
`(mg/ml)
`0.012
`2.56
`
`Dose
`(mg/day)
`9
`20*
`
`Humana
`Ratb
`
`Fold
`Human
`Conc
`
`47-
`520x
`
`*Rat dose: 100 mg/kg/day; body weight 200 g
`a: Aliment Pharmacol Ther 2005; 22: 971-979
`b: J Pharmacy and Pharmacology 2008, 60: 63-70
`
`2. If the sponsor plans histological evaluation of tissues from only control and high dose
`treatment groups, they will also need to conduct histopathologic examination of other dose
`groups under any of the following circumstances:
`(a) for any macroscopic findings in the low and mid dose groups for a. given tissue, they will
`
`
`
`Reference ID: 4000392
`
`213
`
`MYLAN EXHIBIT - 1069 (Part 2)
`Mylan Pharmaceuticals, Inc. v. Bausch Health Ireland, Ltd.
`IPR2022-00722
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
`need to look at that tissue for all of the dose groups
`(b) for statistically significant or otherwise remarkable fmdings in the high dose group, the
`sponsor will need to look at the affected tissues in all of the dose groups.
`(c) for an increase in tumors in an organ for a tumor type that should be analyzed across tissue
`sites as well as by tissue site (e.g., hemangiosarcoma, lymphoma etc.; see McConnell et al,
`JNCI 76:283, 1986) they should look at all relevant tissues for that dose level and the next
`lower dose level,
`(d) for an excessive decrease in body weight or survival in the examined dose group, they
`should examine lower dose groups.
`
`3. Ceaun and stomach should be examined microscopically in all study groups, since these are
`likely to be major sites of pharmacological activity for plecanatide. The Sponsor may
`reconsider their proposed microscopic examination of larynx in all groups, given that this
`tissue is not expected to be a major site of pharmacological activity.
`
`David Jacobson-Kram, Ph.D.
`Chair, Executive CAC
`
`cc:\
`/IND 74,883/Division File, DGIEP
`/David Joseph/Team leader, DGIEP
`/Yuk-Chow Ng/Pharmacologist, DGIEP
`/Matthew Scherer/PM, DGIEP
`/ASeifried, OND IO
`
`
`
`Reference ID: 4000392
`
`214
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
`Executive CAC
`Date of Meeting: January 29, 2013
`
`Committee: Abby Jacobs, Ph.D., OND IO, Acting Chair
`Paul Brown, Ph.D., OND IO, Member
`Tom Papoian, Ph.D., DCRP, Alternate Member
`David Joseph, Ph.D., DGlEP, Team Leader
`Yuk-Chow Ng, Ph.D., DGIEP, Presenting Reviewer
`
`Author of Draft: Yuk-Chow Ng, Ph.D.
`
`The following information reflects a brief summary of the Committee discussion and its
`recommendations.
`
`The Committee did not address the sponsor's proposed statistical evaluation for the
`carcinogenicity bioassay, as this does not affect the sponsor's ability to initiate the bioassay. The
`sponsor may seek guidance on the statistical evaluation of bioassay results from agency staff
`separately. Data files should be submitted electronically following the CDER/CBER Guidance
`for Industry, Providing Regulatory Submission in Electronic Format- Human Pharmaceutical
`Product Applications and Related Submissions Using the eCTD Specifications (June 2008) and
`the associated Study Data Specifications document.
`
`IND #74,883
`
`Drug Name: plecanatide/SP-304
`
`Sponsor: Synergy Pharmaceuticals, Inc.
`
`Background:
`
`The Sponsor is developing plecanatide for the treatment of chronic constipation and
`constipation-predominant irritable bowel syndrome. Plecanatide is a synthetic 16-amino acid
`peptide that is structurally related to the uroguanylin, a member of the guanylin peptide family.
`It is a guanylate cyclase-C (GC-C) agonist. Binding of plecanatide to GC-C increases secretion
`of chloride, bicarbonate, and fluid, through local actions in the intestinal mucosa. Oral
`bioavailability of plecanatide is extremely limited.
`
`Mouse Carcinogenicity Study Protocol and Dose Selection:
`
`The Sponsor proposes a 2-year carcinogenicity study in Albino Crl:CD-1® (ICR) mice
`(b) (4)
`(60/sex/group) at dose levels of 0 (distilled water as vehicle control)11130, or 90 mg/kg/day
`plecanatide, administered by oral gavage.
`
`The Sponsor's dose selection is based on high multiples of the anticipated high dose in humans
`adjusted for body surface area (BSA). The proposed high dose of 90 mg/kg, or 270 mg/m2, is
`
`
`
`Reference ID: 4000392
`
`215
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
`49-fold higher than the predicted maximum human dose of 9 mg/day (5.55 mg/m2 based on a 60-
`kg bodyweight), on a body surface area basis. Plecanatide was tolerated at doses of 20, 60, 150
`and 400 mg/kg in a 26-week study in CD-1 mice. It is noted that systemic exposure of
`plecanatide in the animal species tested has been found to be very low after oral administration,
`which is suggestive of limited oral bioavailabiiity. In the 26-week oral dose range-finding study
`in mice, plecanatide AUC could not be estimated in the 20 or 60 mg/kg/day mice on day 1, or in
`the 20 mg/kg/day mice at week 26, due to the absence of detectable drug levels at most time-
`points. In humans, plecanatide was not detectable in plasma (10 ng/ml limit of quantification) in
`any of the clinical studies. Thus, in this particular situation, where the human AUC can not be
`measured due to the absence of detectable plasma drug levels, the rodent to human AUC ratio
`can not be used for dose selection. The S i onsor provided no specific explanation for the
`(b)
`proposed middle and low dose of 30 and
` g/kg/day, respectively.
`(4)
`
`Executive CAC Recommendations and Conclusions:
`
`1. The Committee recommended doses of 0, 10, 30, and 90 mg/kg/day, by oral gavage.
`The Committee notes that the high dose represents a sufficiently high multiple of local
`(intestinal) exposure relative to a pharmacologically active dose in a mouse model of DSS-
`induced colitis, based on a mg/kg comparison.
`
`2. The Committee noted that the skin infection seen in several of the animals confounded
`interpretation of the 26-week study.
`
`3. The Committee notes that the carcinogenicity study will be performed in a testing facility
`different from the facility that conducted the 26-week dose range-finding study, and a
`different animal vendor may be used as well. Therefore, concurrence on doses is contingent
`on comparable toxicity being achieved in the carcinogenicity study at similar time-points as
`that seen in the 26-week toxicity study.
`
`4. In the event of adverse effects that raise a concern about increased mortality, the Sponsor
`should contact the FDA to obtain concurrence with any proposed change in dosing or
`termination of dosing in a study group prior to initiating the change.
`
`5. If the sponsor plans histological evaluation of tissues from only control and high dose
`treatment groups, they will also need to conduct histopathologic examination of other dose
`groups under any of the following circumstances:
`(a) for any macroscopic findings in the low and mid dose groups for a given tissue, they will
`need to look at that tissue for all of the dose groups
`(b) for statistically significant or otherwise remarkable findings in the high dose group, the
`sponsor will need to look at the affected tissues in all of the dose groups.
`(c) for an increase in tumors in an organ for a tumor type that should be analyzed across tissue
`sites as well as by tissue site (e.g., hemangiosarcoma, lymphoma etc.; see McConnell et al,
`JNCI 76:283, 1986) they should look at all relevant tissues for that dose level and the next
`lower dose level,
`
`
`
`Reference ID: 4000392
`
`216
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
`(d) for an excessive decrea.se in body weight or survival in the examined dose group, they
`should examine lower dose groups.
`
`Abigail Jacobs, Ph.D.
`Acting Chair, Executive CAC
`
`cc:\
`/IND 74,883/Division File, DGIEP
`/David Joseph/Team leader, DGIEP
`/Yuk-Chow Ng/Pharmacologist, DGIEP
`/Matthew Scherer/PM, DGIEP
`/ASeifried, OND IO
`
`
`
`THE FOLLOWING IS THE EXECUTIVE CAC MEETING MINUTES FROM THE FINAL
`REPORT:
`
`
`
`
`Reference ID: 4000392
`
`217
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
`Executive CAC
`Date of Meeting: July 26, 2016
`
`Committee: Karen Davis Bruno, PhD, OND 1O, Chair
`Paul Brown, PhD, OND 1O, Member
`Tim McGovern, PhD, OND 10, Member
`David B. Joseph, PhD, DGIEP, Lead Pharmacologist
`Yuk-Chow Ng, PhD, Presenting Reviewer
`
`Author of Draft: Yuk-Chow Ng, Ph.D.
`
`The following information reflects a brief summary of the Committee discussion
`and its recommendations.
`
`NDA# 208,745
`
`Drug Name: Plecanatide (SP-304)
`
`Sponsor: Synergy Pharmaceutical Inc.
`
`Background:
`
`Plecanatide is a 16-amino acid peptide that binds to guanylate cyclase-C and
`stimulates the production of cyclic guanosine 3',5'-monophosphate. Plecanatide
`is a gastrointestinal prokinetic drug that acts through stimulation of guanylate
`cyclase C in the intestinal mucosa, leading to increased secretion of intestinal
`fluid, and accelerated gastrointestinal transit. Plecanatide is under development
`for treatment of chronic idiopathic constipation.
`
`Plecanatide was negative in the Ames assay, the in vitro L5178Y/TK+/- mouse
`lymphoma mutation assay, and the in vivo mouse bone marrow micronucleus
`assay.
`
`As part of the nonclinical program, the Sponsor conducted a 2-year oral gavage
`carcinogenicity study in mice and a 2-year oral gavage carcinogenicity study in
`rats.
`
`Mouse Carcinogenicity Study:
`
`The Executive CAC recommendations (see meeting minutes dated 1/29/2013)
`for both male and female mice, were doses of 0, 10, 30, and 90 mg/kg/day by
`oral gavage. These doses were based on a sufficiently high multiple of local
`(intestinal) drug concentration relative to a pharmacologically active dose in a
`mouse model of DSS-induced colitis, as predicted by a mg/kg comparison of the
`high dose to the pharmacological dose in mice.
`
`
`
`Reference ID: 4000392
`
`218
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
`In the 104-week oral carcinogenicity study in Crl:[CD-1(ICR)BR] mice, males and
`females were administered 0 (vehicle), 10, 30, or 90 mg/kg/day plecanatide by
`oral gavage. The vehicle was water. Due to low survival in the control males
`and 10 mg/kg/day females, all surviving male and female animals were sacrificed
`beginning on week 98 (males) and 104 (females), respectively, based on the
`Executive CAC recommendations conveyed on November 18, 2014.
`
`There were no significant neoplasms in the mouse study.
`
`Rat Carcinogenicity Study:
`
`The doses tested were in accordance with the Executive CAC recommendations
`(see meeting minutes dated 4/9/2013). For male rats, the Committee
`recommended doses of 0, 10, 30, and 100 mg/kg/day based on the large
`estimated rat to human multiple of local drug concentration in the intestinal tract,
`and the expectation that the local drug concentration in rats will achieve a
`maximum pharmacological effect. For female rats, the Committee recommended
`doses of 0, 10, 30, and 100 mg/kg/day based on reduced bodyweight gain in
`females at 300 mg/kg/day in the dose-ranging study.
`
`In the 104-week oral carcinogenicity study in CD[Crl:CD(SD)] rats, males and
`females were administered 0 (vehicle), 10, 30, or 100 mg/kg/day plecanatide by
`oral gavage. The vehicle was water. The study was terminated on week 94 for
`males and females due to low survival in the control groups, in accordance with
`Executive CAC recommendations conveyed on 2/2/2015.
`
`There were no significant neoplasms in the rat study.
`
`Executive CAC Recommendations and Conclusions:
`
`Mouse:
`
`1. The Committee concluded that the study was adequate, noting prior Exec
`CAC review of the protocol.
`
`2. The Committee concluded that there were no treatment-related neoplasms.
`
`Rat:
`
`1. The Committee concluded that the study was adequate, noting prior Exec
`CAC review of the protocol.
`
`2. The Committee concluded that there were no treatment-related neoplasms.
`
`
`
`Reference ID: 4000392
`
`219
`
`

`

`
`
`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
`Karen Davis Bruno, PhD
`Chair, Executive CAC
`
`cc:\
`/NDA 208,745/Division File, DGIEP
`/David Joseph/Team leader, DGIEP
`/Yuk-Chow Ng/Reviewer, DGIEP
`/Maureen Dewey/PM, DGIEP
`/ASeifried, OND 1O
`
` 9
`
` Reproductive and Developmental Toxicology
`9.1 Fertility and Early Embryonic Development
`Study title: Study of Fertility and Early Embryonic Development to
`Implantation of Plecanatide by Oral Gavage in Mice
`
`
`20016090
`Study no.:
`Study report location: N/A
`Conducting laboratory and location:
`
`(b) (4)
`
`7/5/2011
`Date of study initiation:
`GLP compliance: Yes
`QA statement: Yes
`Drug, lot #, and % purity: Plecanatide (SP-304), batch #101221
`(98.2%)
`
`
`Key Study Findings
` Male mice (25/group) were administered 0 (vehicle), 20, 200, or 600 mg/kg/day
`plecanatide by oral gavage beginning 28 days prior to mating, during mating, and
`until necropsy on dosing days 63 to 66. Plecanatide-treated females were
`treated for at least 14 days prior to mating, during mating, and through gestation
`day (GD) 7. Twenty five mice/sex/group remained untreated throughout the
`study. Each drug-treated male was mated with an untreated female, and each
`female in the drug-treated group was assigned to mate with an untreated male.
`Females (untreated and treated) with a copulatory plug in situ or spermatozoa
`present in a smear of the vaginal contents were considered to be at GD 0, and
`assigned to individual housing. Males and females were sacrificed on dosing
`days 63 to 66 and GD 13, respectively.
`
` There were no meaningful drug-related changes in bodyweight or bodyweight
`gain in males or females.
`
`
`Reference ID: 4000392
`
`220
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
`
`
`In males, there were no drug-related changes in sperm motility or density. In
`females, there were no drug-related effects on estrous cycle, percentage
`pregnancy rate (total pregnant/number mated), the number of corpora lutea,
`implantation sites, live or dead embryos, pre- or post-implantation losses, or
`dams with all nonviable embryos.
`
` The NOAEL for paternal and maternal toxicity is considered to be 600 mg/kg/day,
`based on the absence of drug-related paternal or maternal findings at this dose.
`The NOAEL for male and female fertility, and early embryonic development is
`considered to be 600 mg/kg/day.
`
`Doses: 0 (vehicle), 20, 200, and 600 mg/kg/day
`Frequency of dosing: once daily
`Dose volume: 10 ml/kg
`Route of administration: oral (gavage)
`Formulation/Vehicle: solution / sterile water
`Species/Strain: Mouse/Crl:CD1(ICR)
`Number/Sex/Group: 25
`Satellite groups: None
`Study design: Plecanatide-treated males were dosed
`beginning 28 days prior to mating, during
`mating, and until necropsy (dosing days 63 to
`66). Plecanatide-treated females were dosed
`for at least 14 days prior to mating, during
`mating, and through GD 7. There were also 25
`untreated mice/sex/group (see Sponsor’s table
`below). Each drug-treated male was mated with
`an untreated female. Males that failed to mate
`with an untreated female within the first 14 days
`of cohabitation were assigned an alternate
`untreated female, and remained in cohabitation
`for up to 3 additional days. Each drug-treated
`female was assigned to mate with an untreated
`male. Females (untreated and treated) with a
`copulatory plug in situ or spermatozoa present
`in a smear of the vaginal contents were
`considered to be at GD 0, and assigned to
`individual housing. Males and females were
`sacrificed on dosing days 63 to 66 and GD 13,
`respectively.
`Deviation from study protocol: There were minor deviations that did not affect
`the quality or integrity of the study.
`
`
`
`Methods
`
`
`
`
`
`
`Reference ID: 4000392
`
`221
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
` Study Design of the Fertility and Early Embryonic Development Study
`
`Group
`No.
`
`1
`
`2
`
`3
`
`4
`
`Test Material
`Sterile Water for
`Injection, USP
`
`SP-304
`
`SP-304
`
`SP-304
`
`Dosage
`Level'
`(mg/kg/day)
`
`0 (Vehicle)
`
`20
`
`200
`
`600
`
`Concentration'
`(mg/mL)
`
`Dosage
`Volume
`(mL/kg)
`
`No. of Treated
`Mice
`M
`
`F
`
`No. of
`Untreated Miceb
`F
`M
`
`0
`
`2.0
`
`20.0
`
`60.0
`
`10
`
`10
`
`10
`
`10
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`25
`
`
`
`
`On the day of arrival (5 days before start of treatment), the animals were approximately
`63 days old, and their bodyweights ranged from 28.3 to 36.2 g in males and 21.8 to 28.2
`g in females.
`
`The Sponsor stated that the high dose selection was based on previous toxicity studies
`in mice, the solubility of plecanatide in water, and the expected range of dose levels in
`humans. In 4-week and 13-week oral toxicity studies of plecanatide in mice, the
`NOAELs were considered to be 200 and 20 mg/kg/day, respectively. In a 26-week oral
`toxicity study in mice, the NOAEL was considered to be 400 mg/kg/day, the highest
`dose tested (Note: This reviewer considered the NOAEL to be 150 mg/kg/day in that
`study based on a significant decrease in absolute reticulocyte count and sciatic nerve
`axonal/myelin degeneration at 400 mg/kg/day plecanatide). The oral gavage route of
`administration was selected because it is the intended route of administration in
`humans. The frequency of administration reflected possible clinical use, and the
`duration of administration was in compliance with the appropriate guideline.
`
`Observations and Results
`Mortality
`All animals were checked at least twice daily for mortality. There were no drug-related
`deaths. A 600 mg/kg/day male was euthanized on day 40 of the study and a control
`female was euthanized on day 3 of presumed gestation due to moribund condition,
`which was attributed to intubation error.
`
`Clinical Signs
`Animals were observed at least weekly during the acclimation period, weekly during the
`pre-dose period, once daily before dose administration for all animals, and once daily
`post-dose for the treated females. Post-dose observations were made between 1 and 2
`hours after dose administration.
`
`There were no meaningful drug-related clinical signs. Sporadic clinical signs were
`observed; however, these are not considered to be drug-related because they were
`
`Reference ID: 4000392
`
`222
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`related to an intubation error and/or lacked dose-dependency. Notable clinical
`observations are summarized in the Sponsor’s tables below.
`
` Clinical Observations (Drug-Treated Male Groups)
`
`GROUP
`TEST MATERIAL
`
`DOSAGE (MG/KG/DAY)a
`
`1
`STERILE WATER FOR
`INJECTION, USP
`0 (VEHICLE)
`
`2
`SP-304
`
`20
`
`3
`SP-304
`
`200
`
`4
`SP-304
`
`600
`
`MAXIMUM POSSIBLE INCIDENCE
`
`1611/ 25
`
`1612/ 25
`
`1613/ 25
`
`1588/ 25
`
`UNSCHEDULED EUTHANASIA
`
`DEHYDRATION - TOTAL
`MILD
`SEVERE
`
`PTOSIS
`
`COLD TO TOUCH
`
`URINE-STAINED ABDOMINAL FUR
`
`LEFT AND RIGHT AXILLARY REGION:
`
`SWOLLEN
`
`NECK: SWOLLEN
`
`HUNCHED POSTURE
`
`DECREASED MOTOR ACTIVITY
`
`0
`
`0/
`0/
`0/
`
`0
`0
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`1/
`1/
`0/
`
`1
`1
`0
`
`1
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0/
`0/
`0/
`
`0/
`0/
`0/
`
`0
`0
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`1
`
`1/
`0/
`1/
`
`1/
`
`1
`0
`lb
`
`lb
`
`1/
`
`lb
`
`1/
`
`lb
`
`1/ lb
`
`1/
`
`lb
`
`1/
`
`lb
`
`1/
`
`lb
`
`lb
`
`0/
`0/
`0/
`0/
`0/
`0/
`0/
`
`0/
`
`0
`
`0
`
`0
`
`0
`
`44/
`
`1
`
`0/
`
`0
`
`1/
`
`1/
`
`lb
`
`1/
`
`lb
`
`1/ lb
`
`0/ 0
`
`0/ 0
`
`0/
`
`0/
`
`0/
`
`0
`
`0
`
`0
`
`
`
`
`
`0/
`0/
`0/
`0/
`0/
`0/
`0/
`0/
`0/
`0/
`0/
`0/
`
`0
`
`0
`
`0
`
`0
`
`0
`
`DYSPNEA
`
`BRADYPNEA
`
`HYPERPNEA
`
`REDUCED FECES
`
`LEFT EYE: CORNEAL OPACITY
`
`1/
`0/
`0/
`0/
`0/ 0
`0/
`0/
`0/
`0/ 0
`0/
`0/
`0/
`
`0
`
`0
`
`0
`
`0
`
`TAIL BENT
`
`18/ 1
`
`32/
`
`1
`
`STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF MICE WITH OBSERVATIONS.
`MAXIMUM POSSIBLE INCIDENCE = (DAYS x MICE)/NUMBER OF MICE EXAMINED PER GROUP
`N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF MICE WITH OBSERVATION
`a. Dosage occurred once daily in the morning on Days 1 through 62, 63, 64 or 65 of the study.
`
`
`b. Clinical sign observed in mouse 1493 prior to scheduled dosage on Day 40 of the study.
`UNSCHEDULED EUTHANASIA
`0
`0
`
`0
`
`TIP OF TAIL: BLACK
`
`TIP OF TAIL MISSING
`
`ALL PAWS AND TAIL:
`
`PALE
`
`0/
`0/
`0/
`
`0
`
`0
`
`0
`
`32/
`
`8/
`4/
`
`1
`
`1
`
`0/
`
`0/
`
`0/
`
`0
`
`0
`
`0
`
`STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF MICE WITH OBSERVATIONS.
`MAXIMUM POSSIBLE INCIDENCE = (DAYS x MICE)/NUMBER OF MICE EXAMINED PER GROUP
`N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF MICE WITH OBSERVATION
`a. Dosage occurred once daily
`in the morning on Days 1 through 62, 63, 64 or 65 of the study.
`
`BEST AVAILABLE
`COPY
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`Reference ID: 4000392
`
`223
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
` Clinical Observations (Drug-Treated Female Groups)
`
`GROUP
`TEST MATERIAL
`
`DOSAGE (MG/KG/DAY)a
`
`UNSCHEDULED EUTHANASIA
`
`1
`STERILE WATER FOR
`INJECTION, USP
`0 (VEHICLE)
`
`2
`SP-304
`
`20
`
`1
`
`3
`SP-304
`
`200
`
`0
`
`4
`SP-304
`
`600
`
`0
`
`PRECOHABITATION (DAY 1 OF STUDY TO THE DAY OF COHABITATION) :
`
`MAXIMUM POSSIBLE INCIDENCE
`
`350/ 25
`
`350/ 25
`
`350/ 25
`
`350/ 25
`
`PRESUMED GESTATION:
`
`NO ADVERSE FINDINGS
`
`MAXIMUM POSSIBLE INCIDENCE
`
`340/ 25
`
`350/ 25
`
`350/ 25
`
`350/ 25
`
`TAIL: SWOLLEN
`
`TAIL: SCAB
`
`DEHYDRATION - TOTAL
`MILD
`SLIGHT
`
`RED PERIVAGINAL SUBSTANCE
`
`DECREASED MOTOR ACTIVITY
`
`LACRIMATION
`
`0/
`
`0/
`
`0
`
`0
`
`L/ 1
`0/ 0
`lb
`1/
`
`0/
`
`0
`
`1/
`
`lb
`
`lb
`
`0/ 0
`
`0/ 0
`
`0/ 0
`0/ 0
`0/
`0
`
`1/
`
`1
`
`0/
`
`0
`
`0/ 0
`
`0/ 0
`
`1
`1/
`1/ 1
`0/
`0
`
`0/
`
`0/
`
`0
`
`0
`
`2/ 1
`
`1/
`
`1
`
`0/ 0
`0/ 0
`0/
`0
`
`0/
`
`0/
`
`0
`
`0
`
`0
`
`LIMITED USE OF RIGHT FORELIMB
`
`HUNCHED POSTURE
`
`RIGHT AXILLARY:
`
`SWOLLEN
`
`HEAD TILT TO THE RIGHT
`
`GASPING
`
`BRADYPNEA
`
`1/
`
`:/
`
`lb
`
`"_ /
`
`lb
`
`Li
`
`lb
`
`1/
`
`lb
`
`1/
`
`lb
`
`1/
`
`lb
`
`0/
`
`0/
`
`0/
`
`0/
`
`0/
`
`0/
`
`0/
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0/
`
`0/
`
`0/
`
`0/
`
`0/
`
`0/
`
`0/
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0/
`
`0/
`
`0/
`
`0/
`
`0/
`
`0/
`
`0/
`
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`BEST
`AVAILABLE
`COPY
`
`STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF MICE WITH OBSERVATIONS.
`MAXIMUM POSSIBLE INCIDENCE = (DAYS x MICE)/NUMBER OF MICE EXAMINED PER GROUP
`N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF MICE WITH OBSERVATION
`a. Dosage occurred on Day 1 of study through Day 7 of presumed gestation.
`b. Clinical
`sign observed in mouse 1717 prior to and/or after scheduled dosage on Day 3 of presumed gestation.
`
`
`Body Weight
`Bodyweights were recorded at least weekly during the acclimation period and pre-dose
`period (drug-treated female groups only), daily before each dose was administered, and
`once daily during the post-dose period (drug-treated female groups only). Bodyweights
`for untreated male and female mice were recorded at least once weekly. A terminal
`bodyweight was recorded prior to sacrifice.
`
`There were no meaningful drug-related changes in bodyweight or bodyweight gain in
`males or females. Sporadic statistically significant changes in bodyweight and
`bodyweight gain were noted. However, these changes are not considered to be drug-
`related because they were transient in nature and/or lacked dose-dependency. The
`Sponsor’s figures below summarize the bodyweights in the plecanatide-treated males
`and females during the study period.
`
`
`
`
`
`
`
`Reference ID: 4000392
`
`224
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
` Bodyweights in Drug-Treated Male Groups
`
`39.0
`
`38.0 -
`
`37.0 -
`
`35.0 -
`
`34.0 -
`
`33.0
`
`1'5
`
`22
`
`36
`28a
`DAY OF STUDY
`
`0 (VEHICLE) MG/KG/DAY
`
`20 (SP-304) MG/KG/DAY
`
`200 (SP-304) MG/KG/DAY
`
`-X-
`
`600 (SP-304) MG/KG/DAY
`
`42
`
`49
`
`56
`
`63
`
`
`
` Bodyweights in Drug-Treated Female Groups
`
`a. Last value recorded before cohabitation.
`b. Terminal body weight.
`
`
`
`44.0
`
`43.0 -
`
`42.0
`
`41.0
`
`40.0
`
`39.0 -
`
`38.0 -
`
`37.0
`
`^ 36.0 -
`
`I- 35.0
`
`O 34.0 -
`
`$33.0=
`
`32.0 -
`
`31.0 =
`
`30.0 -
`
`29.0 -
`
`28.0
`
`27.0 -
`
`26.0 -
`
`25.0
`
`0 (VEHICLE) MG/KG/DAY
`
`20 (SP-304) MG/KG/DAY
`
`200 (SP-304) MG/KG/DAY
`
`- X -
`
`600 (SP-304) MG/KG/DAY
`
`140
`
`7
`
`10
`
`13
`
`DAY OF STUDY
`
`DAY OF GESTATION
`
`a. Last value recorded before cohabitation.
`
`
`
`
`
`Feed Consumption
`Not measured.
`
`Estrous Cycles
`
`Estrous cycling was evaluated by examining vaginal cytology in the treated females for
`15 days before the dosing period, 13 days during the dosing period, and then until a
`copulatory plug was observed in situ or spermatozoa was observed in a smear of the
`vaginal contents during the mating period.
`
`There were no drug-related changes in estrous cycles. The Sponsor’s table below
`summarizes the changes in estrous cycles in the plecanatide-treated females.
`
`
`Reference ID: 4000392
`
`225
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
` Effects of Plecanatide on Estrous Cycling (Drug-Treated Female Groups)
`
`GROUP
`TEST MATERIAL
`
`DOSAGE (MG/KG/DAY)a
`
`ESTROUS CYCLING OBSERVATIONS
`
`1
`STERILE WATER FOR
`INJECTION, USP
`0 (VEHICLE)
`
`MICE EVALUATED
`
`N
`
`25
`
`PREDOSAGE ESTROUS CYCLING
`
`2
`SP-304
`
`20
`
`25
`
`3
`SP-304
`
`200
`
`4
`SP-304
`
`600
`
`25
`
`25
`
`ESTROUS STAGES/
`14 DAYS
`
`MICE WITH 6 OR MORE
`CONSECUTIVE
`DAYS OF DIESTRUS
`
`MICE WITH 6 OR MORE
`CONSECUTIVE
`DAYS OF ESTRUS
`
`MEAN±S.D.
`
`2.6 ± 0.8
`
`2.2 ± 0.8
`
`2.7 ± 0.6
`
`2.4 ± 0.6
`
`N(%)
`
`0( 0.0)
`
`1(
`
`4.0)
`
`1( 4.0)
`
`0( 0.0)
`
`N(%)
`
`0( 0.0)
`
`0( 0.0)
`
`1( 4.0)
`
`1( 4.0)
`
`PRECOHABITATION ESTROUS CYCLING b
`
`ESTROUS STAGES/
`14 DAYS
`
`MICE WITH 6 OR MORE
`CONSECUTIVE
`DAYS OF DIESTRUS
`
`MICE WITH 6 OR MORE
`CONSECUTIVE
`DAYS OF ESTRUS
`
`MEAN±S.D.
`
`2.6 ± 0.7
`
`3.0 ± 0.8
`
`2.6 ± 0.7
`
`2.5 ± 0.8
`
`N(%)
`
`3( 12.0)
`
`1(
`
`4.0)
`
`5( 20.0)
`
`3( 12.0)
`
`N(%)
`
`1(
`
`4.0)
`
`0( 0.0)
`
`0( 0.0)
`
`1( 4.0)
`
`through Day 7 of presumed gestation.
`a. Dosage occurred on Day 1 of study
`b. Precohabitation
`period smears obtained on 27 July 2011 were obtained prior
`
`to dose administration.
`
`
`
`I
`
`BEST
`N
`AVAILABLE
`COPY
`
`
`Toxicokinetics
`Not performed.
`
`Dosing Solution Analysis
`The concentration determination of plecanatide in the dosing formulation samples was
`performed in accordance with the validated method. Analyses conducted during the
`treatment period showed that the concentrations of dosing formulations at 2, 20, and 60
`mg/ml were within ±10% of the nominal concentration (+1.1% to +6.5%).
`
`Necropsy
`Males and females were sacrificed on dosing day 63 to 66 and GD 13, respectively, and
`necropsies were performed. The tissues and organs that were collected and weighed
`are summarized in the Sponsor’s table below.
`
`
`
`
`
`
`
`
`
`
`
`Reference ID: 4000392
`
`226
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
`Tissue and Organs Collected and Weighed
`
`Comment
`
`All mice with gross lesions and/or retained tissues.
`
`All female mice at scheduled euthanasia. All female mice terminated early.
`Collected with uterus.
`All treated male mice at scheduled euthanasia. All treated male mice
`terminated early. Individual weight.
`All treated male mice at scheduled euthanasia. All treated male mice
`terminated early.
`Infused with 10% neutral buffered formalin. All treated mice terminated
`early.
`All treated male mice at scheduled euthanasia. All treated male mice
`terminated early.
`All treated male mice at scheduled euthanasia. All treated male mice
`terminated early. Paired weight with and without fluid.
`All mice at scheduled euthanasia. All treated mice terminated early.
`Excluded confirmation of persistent adverse clinical signs.
`All treated mice terminated early.
`All treated mice terminated early.
`All treated mice terminated early.
`Infused with 10% neutral buffered formalin. All treated mice terminated
`early.
`All female mice at scheduled euthanasia. All treated female mice terminated
`early.
`All female mice at scheduled euthanasia. All treated female mice terminated
`early.
`All mice at scheduled euthanasia. All mice terminated early
`All treated mice terminated early.
`All treated mice terminated early.
`All treated male mice at scheduled euthanasia. Treated male mouse
`terminated early. Individual weight; fixed in Bouin's solution for 48 to
`96 hours and retained in neutral buffered 10% formalin.
`
`All mice at scheduled euthanasia. All mice terminated early.
`
`Infused with 10% neutral buffered formalin. All treated mice terminated
`early.
`All female mice at scheduled euthanasia. Treated female mice terminated
`early. Collected with cervix.
`
`
`
`Tissue
`Animal
`identification
`
`Cervix
`
`Epididymides
`
`Epididymis,
`left cauda
`
`Esophagus
`
`Gland, prostate
`
`Gland, seminal
`vesicles
`Gross
`lesions/masses
`Heart
`Kidney
`Liver
`
`Lung
`
`Ovaries
`
`Oviducts
`
`Pituitary
`Spleen
`Stomach
`
`Testes
`
`Tibiofemoral
`joints
`
`Trachea
`
`UteUterus
`
`Weigh Collect
`
`Microscopic
`Evaluation
`
`-
`
`X
`
`X
`
`X
`
`X
`
`X
`
`-
`
`-
`
`X
`
`X
`
`X
`
`X
`
`X
`
`X
`
`X
`
`X
`X
`X
`X
`
`X
`
`X
`
`X
`
`X
`X
`X
`
`X
`
`X
`
`X
`
`X
`
`-
`
`-
`
`-
`
`-
`
`-
`
`X = procedure to be conducted; - = not applicable.
`
`
`
`There were no meaningful drug-related organ weight changes or macroscopic findings.
`The tables below (taken from the sponsor’s study report) summarize organ weights and
`bodyweights in drug-treated males and notable macroscopic findings in drug-treated
`males and females.
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`Reference ID: 4000392
`
`227
`
`

`

`NDA 208,745 Reviewer: Yuk-Chow Ng, PhD
`
`
` Organ Weight and Terminal Body Weight (Drug-Treated Male Groups)
`
`GROUP
`TEST MATERIAL
`
`DOSAGE (MG/KG/DAY)a
`
`MICE TESTED
`
`INCLUDED IN ANALYSES
`
`1
`STERILE WATER FOR
`INJECTION, USP
`0 (VEHICLE)
`
`N
`
`N
`
`25
`
`25