Tab1e 14. Reproductive and Deve10pmenta1 Toxicity: Effects on Pre‑and Postnata1 Deve10pment， Including Materna1 Function (Pivota1) TestArticle: AMG 162
Reproductive and Deve10pmenta1 Toxicity ‑Effects on Pre‑and Postnata1 Deve10pment， Including Materna1 Function
Sp巴cies/Strain: Cynom01gus Monkey Initia1 Age: 4.3 ‑8.6 years of age Date ofFirst D附
・ E ・
2掴
Report Title: Enhanced Pre‑Postnata1 Toxicity Study of AMG 162 Administered by Subcutaneous Injection to Pregnant Cynom01gus Monkeys With up to 6‑Months Postnata1 Eva1uation
Duration ofDosing: 6 months (month1y， from GD Amgen Study No. 112197 20 ‑22 through parturition， up to 6 doses)
Day of Conception: Gestation day 0 Day of Birth: Birth day 1
Method of Administration: Subcutaneous， month1y
Charles River Laboratories PCS‑NV Study No. FIA00588 Specia1 Features: Dams: TK， antibody ana1ysisヲbonebiomarkersヲ Vehicle/Formu1ation: 10 m M sodium acetate， 5% GLP Compliance: Yes mammary g1and deve10pment. sorbit01， pH 5.2
Infants: TKヲantibodyana1ysis， immunophenotyping， bone biomarkersヲ
growth and deve10pment endpoints
No Observed Adverse Effect Level: Not identified for any generation Dos巴(mg/kg/dose)
Number of pregnant fema1esa/live infants
Dams Toxicokinetics: AUC(川 (μg.day/mL) GD 20 ‑22 (白rstdose)
GD 132 (5出dose)
Antibody Ana1ysis: % (incidence) Binding Antibodies
Neutralizing Antibodies
o
(con仕01) 50 29/22b 29116CN A 7400
N A 8620
7 (2/29) 72 (21129) 3 (1/29) 10 (3/29)
Pag巴1of 13 ADA = anti‑drug antibody; AUC = ar巳aund巴rthe conc巴n仕 組onltim巴curv巳 企omtime 0 to tim巴tヲBD=bi巾 day;BMC = bone minera1 content; BMD = bone mineral d巴nsltyヲGD= g巴stationday; PCS‑NV = Pr巴clinicalServices， Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrat巴‑resistantacid phosphatase activ巴isoform;* = p三0.05，*前=pS 0.01，叫*二pS 0.001 using Dunnett's/Dunn's. P1ease see the 1ast page ofthis tab1巴for1ett巴redfootnotes
2.6.7 毒性試験概要表 デノスマブ
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Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Infants Antibody Analysis: % (incidence)
Binding Antibodies 5 (1/21) 50 (7/14)
Neutralizing Antibodies 5 (1/21) 7 (1/14)
Noteworthy Findings
Dams Mortalityd 0 1
Body Weight (Gestation and Postpartum) -
-Food Consumption (Gestation and Postpartum) -
-Clinical Observations (Gestation and Postpartum)
Dystocia - 1
Lethargic, decreased, activity, hunched appearance, poor coordination; secondary to hypocalcemia
- 1
Oral Cavity and Teeth -
-% fetal loss (incidence)
Total fetal loss 24.1 (7/29) 40.7 (11/27)e
1st trimester (GD 20 - 50) 10.3 (3/29) 17.2 (5/29)
3rd trimester (> GD 100) 13.8 (4/29) 22.2 (6/27)e
Stillbirths (> GD 140) 10.3 (3/29) 18.5 (5/27)e
Page 2 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 using Dunnett’s/Dunn’s. Please see the last page of this table for lettered footnotes.
2.6.7 毒性試験概要表 デノスマブ
Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Dams Bone Turnover Biomarkers Osteocalcin (ng/mL)
Predose 28.796 32.984
GD 76 20.123 7.711*
GD 139 16.465 4.428*
PPD 1 10.272 2.660*
PPD 91 43.264 30.347
PPD 180 36.542 52.385*
Bone TRAP 5b (U/L)
Predose 6.286 6.106
GD 76 11.288 4.480*
GD 139 13.937 8.619*
PPD 1 11.010 5.014
PPD 91 6.318 5.568
PPD 180 5.105 8.430
C-terminal Telopeptide (ng/mL)
Predose 1.552 1.541
GD 76 1.604 0.372*
GD 139 1.081 0.244*
PPD 1 1.605 0.509*
PPD 91 2.559 2.566
PPD 180 2.317 2.857
Page 3 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤
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Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Dams Bone Turnover Biomarkers, continued Bone-specific alkaline phosphatase (U/L)
Predose 74.789 72.383
GD 76 63.177 29.079*
GD 139 94.577 53.551*
PPD 1 142.098 61.548
PPD 91 128.570 97.432
PPD 180 112.865 147.537*
Clinical Pathology
Hematology -
-Coagulation -
-Serum chemistry -
-Alkaline phosphastase, GD 139 (U/L) 174 119*
Urinalysis -
-Gross pathology -
-Organ weights -
-Mammary Gland Development -
-Infants Mortality % (incidence) 9.1 (2/22) 50 (8/16)
Maternal rejection f 1 (BD 7) 2 (BD 1, 3)
Infant found dead (unknown cause) 0 1 (BD 11)
Page 4 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 using Dunnett’s/Dunn’s. Please see the last page of this table for lettered footnotes.
2.6.7 毒性試験概要表 デノスマブ
Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Infants Mortality % (incidence), continued
Infant moribund sacrifice 1 (BD 5) 5 (BD 14, 31, 31, 60, 69)
Lip laceration/compromised nursing 1 (BD 5) 0
Fracture/compromised nursing 0 1 (BD 31)
Bacterial sepsis 0 2 (BD 14, 31)
Anemia 0 2 (BD 60, 69)
Body weight (g)
BD 1 340 347
BD 28 410 360
BD 91 717 571*
BD 175 1105 903*
Body weight gain (g)g
BD 1 - 14 26.5 3.7
BD 1 - 112 493.4 317.3*
BD 1 - 175 753.2 605.2*
BD 112 - 175 259.8 287.9
Clinical observations (infants surviving to BD 180)
Swollen digit, left foot 0 1
Tooth eruption -
-Neonatal muscle tone and neurobehavioral assessments -
-Page 5 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤
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Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Infants Immunophenotyping -
-Bone turnover biomarkers Osteocalcin (ng/mL)
BD 1 78.475 59.094
BD 14 188.418 88.728*
BD 91 123.951 198.763
BD 180 86.960 116.841*
Bone TRAP 5b (U/L)
BD 1 37.596 0.809
BD 14 47.228 2.428*
BD 91 64.335 53.954
BD 180 31.769 50.478
C-terminal Telopeptide (ng/mL)
BD 1 1.798 0.568*
BD 14 1.562 0.762*
BD 91 1.282 1.774
BD 180 1.607 1.509
Bone-specific alkaline phosphatase (U/L)
BD 1 275.367 99.838*
BD 14 586.447 481.953
BD 91 750.554 1468.260
BD 180 665.744 1071.855
Page 6 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 using Dunnett’s/Dunn’s. Please see the last page of this table for lettered footnotes.
2.6.7 毒性試験概要表 デノスマブ
Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Infants Clinical Pathology
Hematology -
-Coagulation -
-Serum chemistry -
-Urinalysis -
-Skeletal Evaluations Right femur length (cm)
BD 28 4.61 4.18*
BD 60 5.25 4.54*
BD 90 5.79 5.02*
BD 180 7.09 6.35*
Right femur diameter midshaft (mm)
BD 28 4.00 3.96
BD 60 4.40 4.27
BD 90 4.72 4.42
BD 180 5.57 5.26
Right femur diameter distal metaphysis (mm)
BD 28 9.10 8.94
BD 60 10.02 9.54
BD 90 10.82 11.68
BD 180 12.66 12.03
Page 7 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 using Dunnett’s/Dunn’s. Please see the last page of this table for lettered footnotes.
2.6.7 毒性試験概要表 デノスマブ
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Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Infants Skeletal Evaluations, continued Lumbar spine length (cm)
BD 28 4.68 4.56
BD 60 5.28 4.92*
BD 90 5.95 5.29*
BD 180 7.46 6.96*
Fracture (incidence)
BD 28 0 2
BD 60 0 3
BD 90 0 0
BD 180 0 0
Increased opacity (incidence)
BD 28 0 7
BD 60 0 5
BD 90 0 2
BD 180 0 0
Page 8 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 using Dunnett’s/Dunn’s. Please see the last page of this table for lettered footnotes.
2.6.7 毒性試験概要表 デノスマブ
Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Infants Densitometry (Left Femur – Metaphysis Site) Total area
BD 1 24.48 32.95**
BD 180 50.89 47.76
Total BMC
BD 1 14.16 21.26***
BD 180 22.98 20.98
Total BMD
BD 1 578.3 647.9*
BD 180 453.1 443.2
Trabecular BMC
BD 1 5.01 8.61***
BD 180 3.71 3.85
Trabecular BMD
BD 1 454.1 585.3**
BD 180 161.3 177.4
Cortical/subcortical BMC
BD 1 9.15 12.65**
BD 180 19.27 17.12
Page 9 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 using Dunnett’s/Dunn’s. Please see the last page of this table for lettered footnotes.
2.6.7 毒性試験概要表 デノスマブ
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Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Infants Densitometry, continued Cortical/subcortical BMD
BD 1 679.9 698.9
BD 180 692.1 661.1
Radiographic evaluations (incidence/no. evaluated) Increased opacity
BD 28 0/15 7/8
BD 60 0/13 5/6
BD 90 0/12 2/4
BD 180 0/11 0/3
Fractures (recent or healing)
BD 28 0/15 2/8
BD 60 0/13 3/6
BD 90 0/12 2/4
BD 180 0/11 0/3
Page 10 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 using Dunnett’s/Dunn’s. Please see the last page of this table for lettered footnotes.
2.6.7 毒性試験概要表 デノスマブ
Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Infants Postmortem – BD 1 (Number of infants)h 6 6
Organ weights -
-Macroscopic findings -
-Microscopic findings (incidence)
Lymph nodes – very small/nearly all absent 0 6
Extramedullary hematopoiesis 0 6
Parathyroid gland – chief cell hyperplasia 0 2
Bone – femur, sternum, tibia
Hyperostosis 0 6
Hypoplasia, osteoclast 0 6
Hypertrophy of the physis 0 6
Decreased marrow space 0 6
Fracture 0 1
Bone - mandible
Hyperostosis 0 6
Hypoplasia, osteoclast 0 6
Dental dysplasia 0 6
Teeth, malalignment 0 6
Dental arch, shortened/straighter 0 6
Page 11 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 using Dunnett’s/Dunn’s. Please see the last page of this table for lettered footnotes.
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Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Infants Postmortem – BD 180 (Number of infants) 11 4
Organ weights -
-Macroscopic findings -
-Microscopic findings (incidence)
Lymph nodes – decreased size/some absent 0 4
Extramedullary hematopoiesis 0 4
Mineralization of multiple tissues 0 1
Bone – femur, sternum, tibia
Hyperostosis 0 0
Hypoplasia, osteoclast 0 0
Hypertrophy of the physis 0 0
Decreased marrow space 0 0
Bone - mandible
Hyperostosis 0 0
Hypoplasia, osteoclast 0 0
Page 12 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 using Dunnett’s/Dunn’s. Please see the last page of this table for lettered footnotes.
2.6.7 毒性試験概要表 デノスマブ
Table 14. Reproductive and Developmental Toxicity: Effects on Pre- and Postnatal Development, Including Maternal Function (Pivotal) Test Article: AMG 162
Reproductive and Developmental Toxicity Amgen Study No. 112197
Dose (mg/kg/dose) 0 (control) 50
Infants Postmortem – BD 180 (Number of infants) 11 4
Microscopic findings (incidence)
Dental dysplasia 0 3
Teeth, malalignment 0 1
Dental arch, shortened/straighter 0 1
Page 13 of 13 ADA = anti-drug antibody; AUC = area under the concentration/time curve from time 0 to time t; BD = birth day; BMC = bone mineral content; BMD = bone mineral density; GD = gestation day; PCS-NV = Preclinical Services, Nevada; PPD = postpartum day; TK = toxicokinetics; TRAP = tartrate-resistant acid phosphatase active isoform; * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001 using Dunnett’s/Dunn’s.
a Includes replacement dams (4 control and 3 AMG 162 dams) for presumed abortions before GD50.
b Of the 22 infants, one underwent unscheduled necropsy. Additionally, one was necropsied on BD 7 and was included in the BD1 cohort. Of the remaining 20 surviving infants, 4 were euthanized as age-matched controls for deaths of the AMG 162 infants, 5 were necropsied as part of the BD1 cohort and the remaining 11 continued on study until the scheduled necropsy at BD 180± 2.
c Of the 16 infants, 6 underwent unscheduled necropsy. Additionally, one was necropsied on BD3 and one was necropsied on BD 12 and these were included in the BD1 necropsy cohort. Of the remaining 8 surviving infants, 4 were necropsied as part of the BD1 cohort and the remaining four continued on study until the scheduled necropsy at BD180 ± 2.
d Death occurred on GD151 (secondary to dystocia)
e Adult females 2511 (GD88 abortion) and 2508 (GD156 stillbirth) were excluded from fetal loss calculations except for first trimester because each female had an ADA response beginning at GD76 with subsequent decrease in pharmacologic effect (bone biomarkers) before fetal loss.
f In controls, one infant was rejected (fatally wounded) by the mother and included in the BD1 cohort. In AMG 162-exposed infants, two were rejected by the mother and included in the BD1 cohort.
g Morphometric measurement differences were consistent with the observed decrease in body weight gain, indicating an overall decrease in growth and development in AMG 162-exposed infants and was considered test article related.
h This cohort included five group 1 (control) infants and four group 2 (50 mg/kg) infants necropsied on BD1; the remaining 3 infants were necropsied at later days postpartum based on circumstances that developed, including maternal rejection (Infants 1016 [group 1, BD7] and 2041 [group 2, BD12]) and planned unscheduled necropsy (Infant 2176 [group 2, BD3]).
2.6.7 毒性試験概要表 デノスマブ
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