Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Read-across:Strontium metal completely dissolves upon contact and during the reaction with water under a strong evolution of gas and an immediate precipitation of a white crystalline solid, presumably strontium hydroxide (Sr(OH)2). The water solubility test of strontium (OECD TG 105) indicates a high dissolution from strontium metal (6.74 g/L at 20°C, determined as dissolved strontium, separated by filtration from undissolved test item and precipitates), a rapid formation of Sr2+ + 2OH- + H2 (g) and a corresponding increasing solution pH to a pH > 13. Strontium ions are highly mobile, occur only in one valence state (2+), i.e. are not oxidized or reduced, and do not form strong complexes with most inorganic and organic ligands (Krupka et al. 1999. EPA 402-R-99-004B; Salminen et al. 2015; Carbonaro and Di Toro. 2007. Geochim Cosmochim Acta 71 3958–3968; Carbonaro et al. 2011. Geochim Cosmochim Acta 75: 2499-2511 and references therein). Thus, it may be assumed that systemic toxicological effects (not local) are related to the strontium ion. Therefore, the assessment of the systemic toxicity of strontium is based on elemental strontium concentrations. Read-across of systemic toxicity data available for soluble strontium substances is applied since the strontium ions determine the toxicological potential of strontium. Read-across from strontium ranelate to strontium is possible since as a first surrogate for bioavailability, the solubility of a test substance in water may be applied. Both substances (strontium ranelate and strontium) are soluble (> 5 g/L). Hence, it can be concluded that adverse effects observed with strontium ranelate are due to the presence of the strontium ion and are relevant for strontium metal.


Male and female fertility was examined in a reproduction toxicity study in male and female Wistar rat with oral administration of dose levels of 500, 750 and 1000 mg/kg bw/d Strontium ranelate. Male and female fertility as well as reproductive performance was not affected at dose level of up to 1000 mg/kg bw/d corresponding to 345 mg/kg bw/d strontium. Females of the female fertility subgroup were treated for 14 days prior to pairing with untreated males and continued throughout pairing and until day 17 of gestation. Males of the subgroup for male fertility assessment were treated for 28 days prior to pairing with untreated females and continued throughout pairing including the day before sacrifice. Treatment of females of this subgroup started after mating on day six of gestation until day 20 of lactation.

Link to relevant study records
Reference
Endpoint:
two-generation reproductive toxicity
Remarks:
based on generations indicated in Effect levels (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1997-09-23 to 1998-04-08
Reliability:
2 (reliable with restrictions)
Justification for type of information:
Strontium metal completely dissolves upon contact and during the reaction with water under a strong evolution of gas and an immediate precipitation of a white crystalline solid, presumably strontium hydroxide (Sr(OH)2). The water solubility test of strontium (OECD TG 105) indicates a high dissolution from strontium metal (6.74 g/L at 20°C, determined as dissolved strontium, separated by filtration from undissolved test item and precipitates), a rapid formation of Sr2+ + 2OH- + H2 (g) and a corresponding increasing solution pH to a pH > 13. Strontium ions are highly mobile, occur only in one valence state (2+), i.e. are not oxidized or reduced, and do not form strong complexes with most inorganic and organic ligands (Krupka et al. 1999. EPA 402-R-99-004B; Salminen et al. 2015; Carbonaro and Di Toro. 2007. Geochim Cosmochim Acta 71 3958–3968; Carbonaro et al. 2011. Geochim Cosmochim Acta 75: 2499-2511 and references therein). Thus, it may be assumed that systemic toxicological effects (not local) are related to the strontium ion. Therefore, the assessment of the systemic toxicity of strontium is based on elemental strontium concentrations. Read-across of systemic toxicity data available for soluble strontium substances is applied since the strontium ions determine the toxicological potential of strontium.
Qualifier:
according to
Guideline:
other: ICH Harmonised Tripartite Guideline - Detection of toxicity to reproduction for medical products, Washington June 24, 1993; ICH Harmonized Tripartite Guideline, Addendum: Toxicity to male fertility, July 1996.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: Iffa Credo, Domaine des Oncins, 69210 Saint-Germain sur L' arbresle, France- Age and weight at study initiation: at the beginning of treatment, during the pre-pairing period, the F0 males were nine weeks old and their body weight ranged from 262.5 to 331.3 g. The F0 females intended for delivery of their offspring were thirteen weeks old at the beginning of the pairing period and their body weight ranged from 215.1 to 305.1 g on Gestation Day zero.- Housing: except within the pairing period, the treated F0 males of subgroups B and D, as well as all females, were housed in individual cages. After delivery, F0 dams were kept with their F1 offspring in the same cage until weaning. After weaning, all F1 animals from the same litter, not selected as breeders, were housed together by sex until sexual maturation. The F1 breeders were regrouped up to three per cage for each sex from weaning to pairing. After successful mating in F1 males remained regrouped up to three per cage, while the F1 females were housed in individual cages.- Cage measurements: lenght=45 cm, width = 30 cm, height = 20 cm- Diet (ad libitum): U.A.R (Usine Alimentation Rationnelle, Epinay-sur-Orge, France) sterilized feed pellets- Water (ad libitum): sterilized drinking water- Acclimation period: about two weeksENVIRONMENTAL CONDITIONS- Temperature: 21 ± 1°C- Humidity: 55 ± 15%- Air changes: 14 to 19 times per hour- Photoperiod (hrs dark / hrs light): 12/12- Dust level at filter outlet: less than 40 000 particles ≥ 0.5 µm per m^2 an no particle > 5 µm
Route of administration:
oral: gavage
Vehicle:
other: hydroxyethylcellulose
Details on exposure:
VEHICLE- Batch no.: EI 922- Physical state: powder- Stability: until April 2000- Dissolved in demineralized water at the concentration of 1% w/vPREPARATION OF DOSING SOLUTIONS:- S 12911-2 was suspended in the vehicle prepared as mentioned above- The concentrations of the various preparations were calculated to allow administration at a constant dose volume of 10 mL/kg bwThe dose volume was calculated on the basis of 10 mL/kg bw (body weight not recorded).
Details on mating procedure:
- M/F ratio per cage: 1:1 ratio (subgroup B and D: males were treated)- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of gestation (GD0)
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Tests of stability and homogeneity, carried out before the start of the study (Ginot, Y.M., 1990)*, showed that preparations of S12911-2 in 1% hydroxyethylcellulose were stable for twenty-six days when stored in stoppered flasks at room temperature, at concentrations spanning those administered.Chemical analysis of the preparations administered during the study showed that measured concentrations were close to the intended values.The pH of the test substance preparations was checked and values were found equal to 7.6 for the first set of preparations.*Reference:- Ginot, Y.M. Technologie Servier. Stability Study no.: PA.R. TOX.G04.R02.12911.01, 1990.Homogeneity Study No.: PA.R. TOX.G02.R02.12911.01, 1990
Duration of treatment / exposure:
F0 animals: Male fertility and parturition of F0 females and pre-/postnatal development of F1 animals:- subgroup B (females): day six of gestation and continued until day twenty of lactation inclusive- subgroup B (males): twenty-eight days prior to pairing with treated females from the same subgroup and continued throughout pairing up to, and including, the day before terminal sacrifice (six days after the treated females started littering)Toxicokinetics of F0 males and F0 females during lactation and F1 pups:- subgroup D (females): day six of gestation and continued up to and including day twelve of lactation.- subgroup D (males): twenty-eight-day pre-pairing period and finished the first day of the pairing period
Frequency of treatment:
F0 animals: daily, seven days a week (each treatment was carried out during the morning)
Details on study schedule:
- The method of reproduction used for F1 animals was identical to that used for F0 animals (nocturnal, monogamous pairing). - Selection of parents from F1 generation when pups were eleven to twelve weeks of age at the start of the pairing period which lasted up to twelve days.
Remarks:
Doses / Concentrations:0, 500, 750 and 1000 mg/kg/dayBasis:actual ingested
No. of animals per sex per dose:
Subgroup B: 25 females/25 males each for the control group and the 3 dose levelsSubgroup D: in each treated group five supplementary animals of each sex made up subgroup D for toxicokinetic investigations (exclusively toxicokinetic)(Groups A and C: see section 7.8.2 Developmental toxicity / teratogenicity)
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:The dose levels were chosen on the basis of the results from previous reproductive and general toxicity studies in the rat with S 12911-2.A study on male fertility, carried out by a laboratory (Müller, W. and Semich, R., 1994)* showed that the reproductive performance were not altered when S 12911-2 was administered by gavage at 825 mg/kg/day for eighty days before pairing with untreated female rats. The test substance was well tolerated. No treatment-related mortality was observed.In a preliminary reproductive toxicity study (Momburg, r. et al., 1992)* female rats were treated with S 12911-2 by gavage at 750 mg/kg/day for fourteen days prior to pairing with untreated male rats, throughout pairing and until Gestation Day twenty or until Lactation Day four. The study results show that S 12911-2 did not interfere with the general condition of the females, their fertility, the implantation process and prenatal development of the offspring, as well as the parturition and the viability of the newborn F1 pups until Lactation Day four. Only mean foetal weight on Gestation Day twoenty was slightly decreased.Subchronic and chronic toxicity studies show that S 12911-2 was well tolerated at 750 mg/kg day when administered by gavage over thirteen weeks to Wistar rats or over twenty-six weeks to Sprague Dawley rats (Bazot, D. and Lupart, M., 1997; Nuttall, J. et al., 1996, respectively)*.On the basis of these results and taking into account the duration of treatment (eight to ten weeks for male rats and five to eight weeks for female rats), the high dose was set at 1000 mg/kg/day. Two lower doses were defined by an arithmetical progression, using a factor of 250. Hence, the doses were 500 and 750 mg/kg/day.The pre-pairing treatment period for F0 males was fixed at twenty eight days, because the results of subchronic and chronic toxicity studies (Müller, W. and Semich, R., 1994; Nuttall, J. et al., 1996)* did not reveal either a treatment-related decrease in the weight of male genital organs, or histopathological modifications of the same organs.*References:- Momburg, R., Legrain, B. and Sterz, H.. Etude d'information du S 12911-02 par voie orale chez le rat Wistar. Biologie Servier Internal Report No.: 2349, 1992.- Müller, W. and Semich, R. S12911 - Oral (Gavage) Fertility Study in the Male Rat. Hazleton -18-RFA Project No.: 303-093, 1994- Bazot, D. and Lupart, M. S 12911-2 Toxicity Study by Repeated Oral Administration for 13 Weeks in Wistar Rats. biologie Servier Internal Report No.: 2123,1997.- Nuttall, J. Kelly, J. Barton, C. and Brown, P. S 12911 - 26 Weeks Oral (Gavage) Chronic Toxicity Study in the Rat. Hazleton-25-UK Project No.: 303-88, 1996.
Positive control:
none
Parental animals: Observations and examinations:
F0 GENERATION:CAGE SIDE OBSERVATIONS: Yes Time schedule: - treated animals intended for toxicokinetic investigations (subgroup D) and untreated males (subgroup D) were subjected to a daily mortality survey only.- treated F0 males from subgroup B were observed once daily during acclimation to detect mortality. During treatment, clinical monitoring was carried out prior to and after treatment each day until the day before the start of terminal sacrifice to get equal numbers of recordings for all males. Then treated males were subjected to a daily mortality survey only until their terminal sacrifice.- treated F0 females from subgroup B were observed daily during acclimation to detect mortality. During treatment periods, clinical monitoring was carried out prior to and after treatment, except during pairing. From the start of the pairing period to effective mating, females from both subgroups were subjected to a daily mortality survey only. After each pairing the supposed date of fertilization of the F0 females was determined by examination of the vaginal smears. During gestation phases without treatment, F0 females were observed at least once daily, and any sign of abortion was recorded. During lactation partial or total litter loss was recorded for F0 females from subgroup B. Clinical signs, recorded for non-pregnant females after effective mating, were excluded from evaluation.DETAILED CLINICAL OBSERVATIONS: NoBODY WEIGHT: YesTime schedule for examinations:1) Before pairing:- treated males of subgroup B were weighed once weekly2) After pairing:- males of subgroup B were weighed once weekly for three weeks, as well as on the day of terminal sacrifice- each female of subgroup B effectively mated was weighed daily during gestation, from day zero to day twenty-one in subgroup B- females of subgroup B, which littered, were weighed daily during lactation, from day zero to day twenty-one inclusiveFOOD CONSUMPTION AND WATER CONSUMPTION: YES (females only)- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: NoThese parameters were calculated for the three weeks of gestation in subgroups B, and the two first weeks of lactation in subgroup B.Five different periods were defined for females of subgroup B as follows:- from day zero to day five of gestation inclusive,- from day six to day thirteen of gestation inclusive,- from day fourteen to the end gestation inclusive, - from day zero to day six of lactation inclusive,- from day seven to day thirteen of lactation inclusive
Oestrous cyclicity (parental animals):
F0 GENERATION:For each female, vaginal smears were recorded every day from the first day of pairing until effective mating to determine the stage of oestrous cycle, and to detect potential adverse effects on the normal cycle.
Sperm parameters (parental animals):
Parameters examined in F0 male parental generations:The testes and epididymis from the treated F0 males of subgroup B were systematically sampled and fixed after recording their organ weights and after sperm analysis 8number and viability of spermatozoa) in the left cauda epididymis.
Litter observations:
F1 GENERATIONSTANDARDISATION OF LITTERS- Performed on day 4 postpartum: noPARAMETERS EXAMINEDThe following parameters were examined in F1 offspring:- number of stillbirths, live births, dead births and cannibalized pups (litter affected by maternal cannibalism were excluded from the calculation of sex-ratio)- clinical signs and mortality: after day zero of lactation the F1 pups were observed once a day until weaning, and each dead pup, either spontaneously or by cannibalism, was recorded- body weight: all live pups from each litter were weighed individually on day zero, four, seven, fourteen and twenty-one of lactation; only the F1 breeders (one F1 male and female pup per litter) were weighed once weekly from week four to week ten postpartumPostnatal development: pre-weaning landmarks of physical development and reflex acquisition were observed for all live F1 pups as follows:- physical tests (incisor eruption on Lactation Day eleven, eye opening on Lactation day fifteen, auditory meatus opening on Lactation Day twenty)- reflex tests (surface righting on Lactation Day six, prehensile traction on Lactation Day thirteen, pupillary reflex on Lactation Day nineteen, auditory startle on Lactation Day twenty)Post-weaning landmarks of development (sexual maturation) were observed for all live F1 pups, and behaviour was assessed for all F1 breeders (one male and one female per litter where possible) as follows:- sexual maturation (cleavage of the balanopreputial gland from day forty postpartum (p.p.), vaginal opening from day thirty p.p.)- behaviour (locomotor activity in an open field at the age of six to seven weeks, learning ability in a water maze at the age of nine to ten weeks, memorizing ability in a water maze at the age of ten to eleven weeks)GROSS EXAMINATION OF DEAD PUPS: YES- F1 pups found dead between day zero and day eight of lactation were fixed in alcohol for an examination of their skeleton- F1 pups which died after eight days of age were subjected to a detailed autopsyF2 GENERATION- on the day of birth (day zero of lactation) the number of live newborn pups, dead newborn pups, stillborn pups and cannibalized pups was recorded.- after day zero of lactation the F2 pups were observed once a day until terminal sacrifice, and each dead pup, either spontaneously or by cannibalism, was recorded. Dead pups were fixed in alcohol for an examination of their skeleton.- F2 pups from each litter were weighed individually on day zero, and three of lactation.
Postmortem examinations (parental animals):
F0 GENERATION:SACRIFICE- males of subgroup D and females of subgroups D were killed by carbon dioxide inhalation after successful mating for males, on Lactation Day thirteen, after the last blood sampling for females of subgroup D. These animals were eliminated without necropsy- F0 males of subgroup B were killed by ether inhalation for detailed autopsy, after littering of their corresponding F0 females from subgroup B or from Gestation Day twenty-seven if the corresponding females did not litter- F0 females of subgroup B, which delivered a litter, were killed by ether inhalation from Lactation Day twenty-one for a detailed autopsy. Those from the same subgroup with a sperm positive vaginal smear, but without delivery, were killed by ether inhalation from twenty-five days later for verification of pregnancy and detailed autopsy. Those from the same subgroup with a sperm negative vaginal smear, at the end of the pairing period, were killed by ether inhalation ten days after the last pairing day and eliminated without necropsy. GROSS NECROPSY (TREATED ANIMALS)- a detailed autopsy was carried out on each treated F0 male and each treated F0 female, except those intended exclusively for toxicokinetic investigations- organs with macroscopic anomalies were sampled and preserved for a possible histopathological evaluation; corresponding organs of sufficient controls were preserved for comparison.- the complete male genital tract (testes, epididymides, prostate, seminal vesicles and deferent ducts), from treated F0 males of subgroup B which did not fertilize their corresponding female, was sampled for light microscopic examinationHISTOPATHOLOGY / ORGAN WEIGHTS- no histopathological examination was carried out on organs with macroscopic anomalies, which were sampled from F0 and F1 animals at terminal sacrifice.Microscopic examination was performed on:- testes and epididymides of high dose and control F0 males- complete genital tract of any F0 or F1 males which did not fertilize their corresponding females- ovaries and oviducts of F0 or F1 females which were not pregnant (no implantation site detected after uterus staining)- right knee joint of ten F1 and F2 males and femalesThe other organs, including the macroscopic anomalies, were kept in fixative without any further investigation.
Postmortem examinations (offspring):
F1 GENERATION AND THEIR F2 LITTERSSACRIFICE- F1 males and females not selected for behaviour testing and breeding, were killed separately by ether inhalation after achievement of sexual maturation of all littermates from the same sex in a litter. They were subjected to a detailed autopsy. When a large majority of F1 females had already been killed, and also the F1 males of approximately half of the litters, it was decided to perform an X-ray photography before autopsy on all remaining F1 males from the treated groups and also on one F1 male from each of the remaining control litters, to examine the ribs, the scapulae, the clavicles, the humerus and the long bones of the forelimbs and hind limbs. - F1 males selected for breeding were killed by ether inhalation for a detailed autopsy, after littering of their corresponding F1 females, should the occasion arise, or after terminal sacrifice of apparently non-pregnant F1 females for verification of pregnancy.- F1 females, which delivered a litter and showed no total litter loss, were killed by ether inhalation between Lactation Day three and five for a detailed autopsy. Their F2 pups were subjected to a macroscopic external examination to detect any anomaly, before killing.- F1 females with a sperm positive vaginal smear, but without delivery, were killed by ether inhalation from twenty-six days later for verification of pregnancy and a detailed autopsy.GROSS NECROPSY- a detailed autopsy was carried out on each F1 male and female at terminal sacrifice, and on all F1 animals found dead during the study if they were aged at least nine days- organs with macroscopic anomalies were sampled and preserved for a possible histopathological evaluation; corresponding organs of sufficient controls were preserved for comparison- the complete male genital tract (testes, epididymides, prostate, seminal vesicles and deferent ducts), from F1 males which did not fertilize their corresponding females, was sampled for light microscopic examination- the ovaries and oviducts were sampled for a possible light microscopic examination from F1 females which were apparently not pregnant at terminal sacrifice- the right knee joint (femoro-tibial joint) of F1 males and females from the first ten litters, with a least one viable F2 male and female pup, was sampled for light microscopic examination- in each of these litters, one F2 male and female were selected at random for the same examination
Statistics:
Please refer to the field "Any other information on materials and methods incl. tables" below.
Reproductive indices:
- gestation index (number of females with a live litter / number of pregnant females)- mean live birth index (number of live newborn pups / number of implantation sites)
Offspring viability indices:
- weaning index- viability index- survival index
Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
not examined
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed
1) F0 GENERATION (F0 males from subgroup B (F0 male general condition and fertility)) CLINICAL SIGNS AND MORTALITY- administration of S 12911-2 to F0 males from subgroup B did not provoke mortality or changes in behaviour; only discolouration of faeces for all animals of all dose groups was seenBODY WEIGHT AND FOOD CONSUMPTION- body weight of F0 males was not adversely affected by the test substance in any of the treated groupsREPRODUCTIVE FUNCTION: SPERM MEASURES- determination of the mean number of spermatozoa per gram cauda epididymis (x10^6) revealed similar values for all groups (not statistically significant)- evaluation of sperm viability (5 motile sperm) per cauda epididymis led to similar values for all groups (not statistically significant)REPRODUCTIVE PERFORMANCE- libido of F0 males was not altered by treatment with S 12911-2 in any dose group- the effective mating rate was similar for all groups- the mean time necessary for successful copulation, expressed in days, was also similar for all groups- the fertility index was similar for F0 males from treated and control groupsORGAN WEIGHTS- reproductive organ weights: absolute mean and relative (% to body weight) mean weights of testes and epididymides were similar between treated and control groupsGROSS PATHOLOGY (PARENTAL ANIMALS)- findings are not attributed to an adverse effect of the test substanceHISTOPATHOLOGY- systemic histomorphological examination of the testes and epididymides in all F0 males from the high dose and control groups, as well as examination of the complete genital tract in those of the F0 males that did not fertilize their corresponding females, did not reveal any anomaly attributable to treatment with the test substance2) F0 GENERATION (F0 females from subgroup B (F0 female parturition and general condition during gestation and lactation))CLINICAL SIGNS AND MORTALITY- administration of S 12911-2 t F0 females of subgroup B, from Gestation Day six to Lactation Day twenty, did not provoke mortality or changes in behaviour; only discolouration of faeces for all animals of all dose groups was seenBODY WEIGHT AND FOOD CONSUMPTION- body weight of F0 females from subgroup B was not adversely affected by the test substance in any of the treated groups- treatment of F0 females from subgroup B with 12911-2 did not interfere with their feed consumption up to the highest dose administeredWATER CONSUMPTION- treatment of F0 females from subgroup B with S 12911-2 did not interfere with their water consumption up to the highest dose administeredGROSS PATHOLOGY- autopsy of the F0 females showed findings which are not attributed to an adverse effect of the test substanceGESTATION and PARTURITION- no interference with normal progress of gestation and parturition- no effects on mean duration of gestation or gestaion index- mean live birth index not affected by treatment- slight differences between treated groups were due to one female in each group mated during metoestrus (without biological relevance)
Dose descriptor:
NOAEL
Remarks:
(general toxicity, male fertility and reproductive performance)
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Remarks on result:
other: Generation: F0 (migrated information)
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
1) F1 pupsOTHER FINDINGS- the sex ratio of F1 pups from treated females was close to 1.00, and therefore, was not modified by the test substanceVIABILITY- S 12911-2 had no impact on F1 pup viability after birth- mortality of F1 pups from birth to weaning was similar in all groups- no litter loss was observed in any of the groups, consequently, the weaning index was similar in all groupsCLINICAL SIGNS (OFFSPRING)- no treatment-related findings were madeBODY WEIGHT- mean body weight of F1 pups, as well as changes in mean body weight of F1 pups, from birth to weaning, were similar for treated and control groupsSEXUAL MATURATION- sexual maturation of F1 animals was not altered by treatment of the animals from the parental F0 generationGROSS PATHOLOGY- the external macroscopic examination of F1 pups at birth did not reveal any abnormalitySkeletal examination of F1 pups found dead before lactation day nine revealed the following findings:- one control pup with one rudimentary rib and a thoracic vertebra limited to the right transverse process- one pup at 750 mg/kg/day with one rudimentary rib and wavy ribs- three pups at 1000 mg/kg/day with wavy ribs- one pup at 1000 mg/kg/day with wavy ribs, one rudimentary rib, a bent scapula and a misshapen clavicle- one pup at 1000 mg/kg/day with a delay in the ossification of two sternebrae and two caudal vertebrae, as well as a misshapen clavicle.- autopsy at terminal sacrifice of the F1 animals not selected for breeding revealed no adverse effect of the test substance- X- ray radiography of F1 males of approximately half of the litters, did not detect any bent, or shortened and thickened, or misshapen bone among the ribs, the scapulae, the clavicles, the humerus and the long bones of the forelimbs and hind limbsOTHER FINDINGSPhysical development:- F1 pups from treated groups showed a percentage of success equal to or higher than that of control pups for eye opening on Lactation Day fifteen (LD 15) and for the auditory meatus opening on LD 20 - the incisor eruption on LD 11 appeared to be delayed for a higher percentage of pups from the treated groups than from the control group, slightly at 500 and 750 mg/kg/day, moderately at 1000 mg/kg/day (3.7, 7.3, 10.6, 27.1% at 0, 500, 750 and 1000 mg/kg/day respectively). The difference from the control group was statistically significant at 750 and 1000 mg/kg/day (p<0.01), but had no consequential negative effect on animal growthReflex acquisition:- similar or identical percentages of successful F1 pups in treated and control groups for, surface righting on LD 6, prehensile traction on LD 13, pupillary reflex on LD 19 and auditory startle on LD 20 - difference between treated and control groups were not statistically significant, except for surface righting at 500 mg/kg/day (73.2% against 63.7% for controls, p<0.05)2) F1 BREEDERSCLINICAL SIGNS- treatment of parental F0 generation had no adverse effect on F1 male and female locomotor activity in an open field whatever the parameter evaluated- treatment of parental F0 generation had also no adverse effect on learning and memorizing abilities of F1 males and females in a water maze; treated animals performed generally better than controlsBODY WEIGHT- mean body weight gain of F1 males and females, randomly selected at weaning (three weeks old) as future breeders, was similar between the third and tenth week pp. for treated and control groups- mean body weights of F1 males and females at the age of ten weeks were not statistically different between treated and control groupsREPRODUCTIVE PERFORMANCELibido: - all F1 males mated their corresponding F1 female- the effective mating rate was 1.00 for all groups- the mean time necessary for successful copulation, expressed in days, was similar for all groupsFertility index and reproductive performances: - the fertility index was not adversely affected- the delivery data for the F1 females on Lactation Day zero revealed no statistically or biologically significant differences between treated and control groups for any of the parameters evaluatedGestation and Parturition:- treatment of females from the parental F0 generation from implantation, through gestation and parturition until weaning, had no impact on the normal progress of gestation of the F1 females- the mean duration of gestation was almost identical for all groups.- the gestation index was slightly higher in all treated than control groups (0.94, 1.00, 0.95, 1.00, at 0, 500, 750 and 1000 mg/kg/day respectively)- the mean live birth index was similar between groups (0.89, 0.88, 0.87, 0.88 at 0, 500, 750 and 1000 mg/kg/day respectively)GROSS PATHOLOGY- autopsy of F1 breeders at terminal sacrifice revealed no findings which were attributed to an adverse effect of the test substance- the systemic histological examination of the femoro-tibial joints of F1 breeders from control and high dose groups did not detect any abnormal finding; adjacent osseous and cartilaginous structures were also normal- the systemic histopathological examination of ovaries and oviducts from non-pregnant females (F1 breeders), as well as that of the complete genital tract from the corresponding males, did not reveal any abnormal findingOTHER FINDINGS (OFFSPRING)- mean body weight of the F2 pups at birth was similar for treated and control groups- the sex-ratio of F2 pups from treated females was within the normal range, and therefore, was not modified by the test substance- the external macroscopic examination of F2 pups revealed only one live newborn control pup with a shortened tail due to maternal cannibalism3) F2 PUPSVIABILITY- treatment of F0 females with S 12911-2, from implantation to weaning of F1 pups, had no impact on F2 pup viability during the early postnatal phase until Lactation Day three - no litter loss was observed in any of the groups- the viability indes was not adversely affectedCLINICAL SIGNS- F2 pups from all treated groups did not show any particular clinical signBODY WEIGHT- mean body weight, as well as changes in mean body weight of F2 pups, from birth to Lactation Day three, were similar for treated and control groupsGROSS PATHOLOGY (OFFSPRING)Examination of F2 pups found dead before terminal sacrifice:- skeletal examination of dead F2 pups resulted in a compression of the right transverse processes of the third to the sixth lumber vertebrae in a F2 control pup- one wavy rib in a F2 pup from the 1000 mg/kg/day dose group is not attributed to any effect of the test substance, because of its spontaneous occurrence in this rat strainExamination of F2 pups at terminal sacrifice- macroscopic external examination of F2 pups at terminal sacrifice (between Lactation Day three and five) revealed no abnormality- systematic histological examination of the femoro-tibial joints of F2 pups from the control and high dose groups did not detect any abnormal finding; adjacent osseous and cartilaginous structures were also normal
Dose descriptor:
NOAEL
Remarks:
(postnatal development)
Generation:
F1
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects on postnatal development up to 1000 mg/kg bw/d corresponding on LD13 to F1 pup strontium plasma concentration of 7.5 µg/mL just after the 24 h time point for dams.
Dose descriptor:
NOAEL
Remarks:
(fertility)
Generation:
F1
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects on male and female fertility
Dose descriptor:
NOAEL
Remarks:
(pre- and early postnatal development)
Generation:
F2
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects on pre- and early postnatal development
Reproductive effects observed:
not specified

TOXICOKINETIC EVALUATION

- after administration of S 12911 -2 to F0 females sugroup D, strontium and ranelate were detected in the plasma of all animals; this proves that the compound was absorbed at each dose level

- it appears that males were more exposed to strontium than non-pregnant females; as dosing of males before pairing covered a period twice as long as that of females (28 versus 14 days), steady state may not have been reached by Day 14

- no gender differences were observed for the systemic exposure to ranelate

- mean accumulation ratios were close to unity, however, it was unclear whether steady state was reached

- systemic exposure (AUC24) of males to strontium and maximal plasma concentrations (Cmax) increased significantly less than dose

- no correlation was found between dose and AUC24- or Cmax-values for ranelate

- AUC24 -values of strontium in females increased proportionally to dose on MD-1 and Lactation Day 12, but significantly less than proportionally to dose on Gestation Day 6

- no correlation was found on Gestation Day 17; AUC24 -values of ranelate in females increased proportionally to dose on MD-1, Gestation Day 6 and gestation Day 17; no correlation was found on Lactation Day 12

- Cmax-values of strontium and ranelate increased significantly less than proportionally to dose on MD-1 and proportionally to dose on Gestation Day 6; no correlation was found on Gestation Day 17 and Lactation Day 12

- it appears that at all dose levels, mean strontium plasma levels measured at about 24 hours post-dosing of dams on Lactation Day 12 were about 2 -fold higher in the untreated F1 pups than in the corresponding F0 dams

- the determination of strontium concentrations in the bone of F1 pups revealed a dose-related increase which was consistent with that observed for strontium plasma levels in F1 pups on Lactation Day 13; consequently, the percentage of strontium in F1 rat bones (calculated as the molar ratio between strontium and the sum of strontium + calcium) also increased with dose, and in a very similar proportion to the strontium plasma levels

Conclusions:
No observed adverse effect levels (NOAELs) of S 12911-2 in the rat:The NOAEL for F0 female general toxicity and reproductive performance is 1000 mg/kg/day, corresponding on LD12 to maternal AUC24 and C24h-values of 441 µg.h/mL and 3.5 µg/ml of strontium, respectively, and 41.4 µg.h/mL and 0.3 µg/ml of ranelate, respectively, and on the last day of the pre-pairing period to AUC24-values for males of 376 µg.h/mL of strontium and 16.5 µg.h/mL of ranelate.The NOAEL for postnatal development of the F1 generation is 1000 mg/kg/day, corresponding on LD13 to F1 pup strontium plasma concentration of 7.5 µg/mL just after the 24 h time point for dams. The lower percentage of pups with incisior eruption was regarded as not adverse because it had no consequential negative effect on growth. The NOAEL for F1 male and female fertility is 1000 mg/kg/day. The NOAEL for pre-and early postnatal development of the F2 generation is 1000 mg/kg/day.
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
345 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The study to examine the effects of strontium ranelate on male and female fertility (K. Momburg, 2001) of rats is regarded as relevant and reliable to evaluate the effects of strontium; i.e., a reliable GLP reproduction toxicity study with strontium ranelate (Oral reproduction toxicity study in the Wistar rat (male and female fertility/embryo-fetal and postnatal development) addressing embryo-fetal and pre-/postnatal development of offspring according to the ICH guideline on Detection of Toxicity to Reproduction of Medicinal Products, June 24, 1993, and with the ICH Guideline Addendum: Toxicity to male fertility, July 1996.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available

Effects on developmental toxicity

Description of key information

Read-across:Strontium metal completely dissolves upon contact and during the reaction with water under a strong evolution of gas and an immediate precipitation of a white crystalline solid, presumably strontium hydroxide (Sr(OH)2). The water solubility test of strontium (OECD TG 105) indicates a high dissolution from strontium metal (6.74 g/L at 20°C, determined as dissolved strontium, separated by filtration from undissolved test item and precipitates), a rapid formation of Sr2+ + 2OH- + H2 (g) and a corresponding increasing solution pH to a pH > 13. Strontium ions are highly mobile, occur only in one valence state (2+), i.e. are not oxidized or reduced, and do not form strong complexes with most inorganic and organic ligands (Krupka et al. 1999. EPA 402-R-99-004B; Salminen et al. 2015; Carbonaro and Di Toro. 2007. Geochim Cosmochim Acta 71 3958–3968; Carbonaro et al. 2011. Geochim Cosmochim Acta 75: 2499-2511 and references therein). Thus, it may be assumed that systemic toxicological effects (not local) are related to the strontium ion. Therefore, the assessment of the systemic toxicity of strontium is based on elemental strontium concentrations. Read-across of systemic toxicity data available for soluble strontium substances is applied since the strontium ions determine the toxicological potential of strontium. Read-across from strontium ranelate and strontium nitrate to strontium is possible since as a first surrogate for bioavailability, the solubility of a test substance in water may be applied. All substances (strontium ranelate, strontium nitrate and strontium) are soluble (> 5 g/L). Hence, it can be concluded that adverse effects observed with strontium ranelate or strontium nitrate are due to the presence of the strontium ion and are relevant for strontium metal. The effects of strontium ranelate on the embryo-fetal and pre-/postnatal development was investigated in a GLP compliant toxicity study according to the ICH guideline on Detection of Toxicity to Reproduction of Medical Products, June 24, 1993 (K. Momberg 2001). Groups of pregnant female rats were either exposed from day 14 prior to mating with untreated males until day 17 of gestation (group A) or after mating with treated males from day 6 of gestation until day 20 of lactation (group B). Embryo-fetal development was evaluated in group A and effects on parturition and pre-/post natal development in group B. Animals received the test compound by oral gavage at dose levels of 500, 750 and 1000 mg/kg bw/d. Control animals were treated with the vehicle. Additional subgroups (group C and D) were included for toxicokinetic investigations. No treatment-related maternal toxicity was observed in group A and B females. No effects on embryo/-fetal development were observed beside an increase in the frequency of delays in skeletal ossification and structural abnormalities (way ribs, bent bones, shortened and thickened humerus and misshapen clavicle) at all dose levels. Although the percentage of affected fetuses by a delay of ossification was higher in the treated groups than in the control, there was no dose-response relationship and values were within the normal historical control range of this strain. In addition, the structural abnormalities observed in twenty day old fetuses were completely reversible in seven to eight week old F1 animals, because all these anomalies were no longer visible during postnatal development as shown by X-ray radiography. These transitory findings were regarded as not effecting basical development of offspring but were related to retarded ossification. It was also discussed that these findings appear not relevant in the case of human exposure during organogenesis, because the skeletal development at parturition in humans is much more advanced than in rodent species. In conclusion, these findings were not considered as true congenital skeletal malformations, because they were reversible and were therefore considered as variations without any functional consequences. Although in the context of the study, the lowest dose level of 500 mg/kg bw/d strontium ranelate corresponding to 172 mg/kg bw/d strontium does not represent an NOAEL, but a LOAEL, the findings at this dose level were of transient nature and regarded as not relevant for human embryo/fetal development, and thus did not provide evidence of an adverse effect on the development of offspring. The structural abnormalities were also not present in theoral embryo-fetal development study with strontium ranelate (K. Momberg 1999) in rabbits at up to 1500 mg/kg bw/d. In the post-natal development part of the study, a delay of incisor eruption was seen on lactation day 11 mainly in offspring of the 1000 mg/kg bw/d group, but had no negative effect on animal growth which is directly related to feed intake and use of teeth for gnawing of feed pellets. In addition, the relevance of these effects for humans was deemed as low, because tooth development in man differ from rat with incisor eruption occurring after weaning at 6-24 months after birth. Therefore, the NOAEL for postnatal development was established at the highest dose group of 1000 mg/kg bw/d strontium ranelate corresponding to 345 mg/kg bw/d strontium. In addition, in the study published by Lansdown et al. (1972) (see 7.8.2 "s_Lansdown_1972) groups of 3 female Wistar rats were treated subcutaneously with 25, 50, 100 or 200 mg strontium nitrate/kg bw/d in 1 ml distilled water from day 9 to 19 of pregnancy when they were killed. Control animals received distilled water only. The progeny from strontium-treated mothers did not differ from that of controls in size or body weight. The litter sizes were normal and the number of resorption sites was not increased. No histological changes were detected in the soft tissues and the skeletal tissues exhibited the characteristic degree of ossification for 19-day old rat fetuses. The results indicate that high doses of strontium nitrate (up to and including 200 mg/kg bw/d s. c.) are not teratogenic. Thus, the dose of 200 mg/kg bw/d can be considered as a NOAEL for developmental toxicity corresponding to an external dose of about 83 mg/kg bw/d strontium to female rats. However, the study was conducted in a small number of females (p=3) per group and a limited number of parameters was evaluated. In addition, the subcutaneous route of administration is not relevant route of exposure for risk assessment purposes. Nevertheless, the study by Lansdown et al. (1972) on pregnant rats is regarded as appropriate to support the evaluation of the effects of strontium on embryo-fetal development.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
1997-09-23 to 1998-04-08
Reliability:
2 (reliable with restrictions)
Justification for type of information:
Strontium metal completely dissolves upon contact and during the reaction with water under a strong evolution of gas and an immediate precipitation of a white crystalline solid, presumably strontium hydroxide (Sr(OH)2). The water solubility test of strontium (OECD TG 105) indicates a high dissolution from strontium metal (6.74 g/L at 20°C, determined as dissolved strontium, separated by filtration from undissolved test item and precipitates), a rapid formation of Sr2+ + 2OH- + H2 (g) and a corresponding increasing solution pH to a pH > 13. Strontium ions are highly mobile, occur only in one valence state (2+), i.e. are not oxidized or reduced, and do not form strong complexes with most inorganic and organic ligands (Krupka et al. 1999. EPA 402-R-99-004B; Salminen et al. 2015; Carbonaro and Di Toro. 2007. Geochim Cosmochim Acta 71 3958–3968; Carbonaro et al. 2011. Geochim Cosmochim Acta 75: 2499-2511 and references therein). Thus, it may be assumed that systemic toxicological effects (not local) are related to the strontium ion. Therefore, the assessment of the systemic toxicity of strontium is based on elemental strontium concentrations. Read-across of systemic toxicity data available for soluble strontium substances is applied since the strontium ions determine the toxicological potential of strontium.
Qualifier:
according to
Guideline:
other: ICH Harmonised Tripartite Guideline S5 - Detection of toxicity to reproduction for medical products to male fertility, Washington June 24, 1993; ICH Harmonized Tripartite Guideline, Addendum: Toxicity to male fertility; July 1996.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS- Source: Iffa Credo, Domaine des Oncins, 69210 Saint-Germain sur L' arbresle, France- Age and weight at study initiation: at the beginning of treatment, during the pre-pairing period, the F0 males were nine weeks old and their body weight ranged from 262.5 to 331.3 g, the F0 females intended for Caesarian section were eleven weeks old and their body weight ranged from 217.9 to 261.2 g. The F0 females intended for delivery of their offspring were thirteen weeks old at the beginning of the pairing period and their body weight ranged from 215.1 to 305.1 g on Gestation Day zero.- Housing: except within the pairing period, the treated F0 males of subgroups B and D, as well as all females, were housed in individual cages. The untreated males of subgroups A and C were regrouped up to three per cages. After delivery, F0 dams were kept with their F1 offspring in the same cage until weaning. After weaning, all F1 animals from the same litter, not selected as breeders, were housed together by sex until sexual maturation. The F1 breeders were regrouped up to three per cage for each sex from weaning to pairing. After successful mating in F1 males remained regrouped up to three per cage, while the F1 females were housed in individual cages. - Cage measurements: length =45 cm, width = 30 cm, height = 20 cm- Diet (ad libitum): U.A.R (Usine Alimentation Rationnelle, Epinay-sur-Orge, France) sterilized feed pellets- Water (ad libitum): sterilized drinking water- Acclimation period: about two weeksENVIRONMENTAL CONDITIONS- Temperature: 21 ± 1°C- Humidity: 55 ± 15%- Air changes: 14 to 19 times per hour- Photoperiod (hrs dark / hrs light): 12/12- Dust level at filter outlet: less than 40 000 particles ≥ 0.5 µm per m^2 an no particle > 5 µm
Route of administration:
oral: gavage
Vehicle:
other: hydroxyethylcellulose
Details on exposure:
VEHICLE- Batch no.: EI 922- Physical state: powder- Stability: until April 2000- Dissolved in demineralized water at the concentration of 1% w/vPREPARATION OF DOSING SOLUTIONS:- S 12911-2 was suspended in the vehicle prepared as mentioned above- The concentrations of the various preparations were calculated to allow administration at a constant dose volume of 10 mL/kg bwThe dose volume was calculated on the basis of 10 mL/kg bw (body weight not recorded).
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Tests of stability and homogeneity, carried out before the start of the study (Ginot, Y.M., 1990)*, showed that preparations of S12911-2 in 1% hydroxyethylcellulose were stable for twenty-six days when stored in stoppered flasks at room temperature, at concentrations spanning those administered.Chemical analysis of the preparations administered during the study showed that measured concentrations were close to the intended values.The pH of the test substance preparations was checked and values were found equal to 7.6 for the first set of preparations.*Reference:- Ginot, Y.M. Technologie Servier. Stability Study no.: PA.R. TOX.G04.R02.12911.01, 1990.Homogeneity Study No.: PA.R. TOX.G02.R02.12911.01, 1990
Details on mating procedure:
- M/F ratio per cage: 1:1 ratio (subgroup B and D: males were treated)- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of gestation (GD0)
Duration of treatment / exposure:
F0 animals: Female fertility and embryofetal development:- subgroup A (females): fourteen days prior to pairing with untreated males from the same subgroup and continued throughout pairing and until day seventeen of gestation inclusiveToxicokinetics during pre-mating and gestation periopds:- subgroup C (females): fourteen days prior to pairing with untreated males from the same subgroup and continued throughout pairing and until day seventeen of gestation inclusive
Frequency of treatment:
F0 animals: daily, seven days a week (each treatment was carried out during the morning)
Duration of test:
Until gestation day 20
No. of animals per sex per dose:
Subgroup A: 20 females each for the control group and the 3 dose levels (males were also included)Subgroup C: in each treated group five supplementary females made up subgroup C for toxicokinetic investigations(Groups B and D: see section 7.8.1 Toxicity to reproduction)
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:The dose levels were chosen on the basis of the results from previous reproductive and general toxicity studies in the rat with S 12911-2.A study on male fertility, carried out by a laboratory (Müller, W. and Semich, R., 1994)* showed that the reproductive performance were not altered when S 12911-2 was administered by gavage at 825 mg/kg/day for eighty days before pairing with untreated female rats. The test substance was well tolerated. No treatment-related mortality was observed.In a preliminary reproductive toxicity study (Momburg, r. et al., 1992)* female rats were treated with S 12911-2 by gavage at 750 mg/kg/day for fourteen days prior to pairing with untreated male rats, throughout pairing and until Gestation Day twenty or until Lactation Day four. The study results showed that S 12911-2 did not interfere with the general condition of the females, their fertility, the implantation process and prenatal development of the offspring, as well as the parturition and the viability of the newborn F1 pups until Lactation Day four. Only mean foetal weight on Gestation Day twoenty was slightly decreased.Subchronic and chronic toxicity studies showed that S 12911-2 was well tolerated at 750 mg/kg day when administered by gavage over thirteen weeks to Wistar rats or over twenty-six weeks to Sprague Dawley rats (Bazot, D. and Lupart, M., 1997; Nuttall, J. et al., 1996, respectively)*.On the basis of these results and taking into account the duration of treatment (eight to ten weeks for male rats and five to eight weeks for female rats), the high dose was set at 1000 mg/kg/day. Two lower doses were defined by an arithmetical progression, using a factor of 250. Hence, the doses were 500 and 750 mg/kg/day.*References:- Momburg, R., Legrain, B. and Sterz, H.. Etude d'information du S 12911-02 par voie orale chez le rat Wistar. Biologie Servier Internal Report No.: 2349, 1992.- Müller, W. and Semich, R. S12911 - Oral (Gavage) Fertility Study in the Male Rat. Hazleton -18-RFA Project No.: 303-093, 1994- Bazot, D. and Lupart, M. S 12911-2 Toxicity Study by Repeated Oral Administration for 13 Weeks in Wistar Rats. biologie Servier Internal Report No.: 2123,1997.- Nuttall, J. Kelly, J. Barton, C. and Brown, P. S 12911 - 26 Weeks Oral (Gavage) Chronic Toxicity Study in the Rat. Hazleton-25-UK Project No.: 303-88, 1996.
Maternal examinations:
F0 GENERATION:CAGE SIDE OBSERVATIONS: Yes Time schedule: - treated animals intended for toxicokinetic investigations (subgroup C) and untreated males (subgroups A) were subjected to a daily mortality survey only.- treated F0 females from subgroup A were observed daily during acclimation to detect mortality. During treatment periods, clinical monitoring was carried out prior to and after treatment, except during pairing. From the start of the pairing period to effective mating, females were subjected to a daily mortality survey only. After each pairing the supposed date of fertilization of the F0 females was determined by examination of the vaginal smears. During gestation phases without treatment, F0 females were observed at least once daily, and any sign of abortion was recorded. DETAILED CLINICAL OBSERVATIONS: NoBODY WEIGHT: YesTime schedule for examinations:1) Before pairing:- treated females of subgroup A were weighed once weekly.2) After pairing:- each female of subgroups A effectively mated was weighed daily during gestation, from day zero to day twenty in subgroup A.FOOD CONSUMPTION AND WATER CONSUMPTION: YES (females only)- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: NoThese parameters were calculated for the two weeks of the premating period in subgroup A, the three weeks of gestation in subgroups A.Six different periods were defined for females of subgroup A as follows:- from day fourteen to day eight inclusive prior to the start of animal pairing, - from day seven to day two inclusive prior to the start of animal pairing,- from day zero to day five of gestation inclusive,- from day six to day eleven of gestation inclusive,- from day twelve to day seventeen of gestation inclusive,- from day eighteen to day nineteen of gestation inclusiveFive different periods were defined for females of subgroup B as follows:OESTRUS CYCLICITYF0 GENERATION:For each female, vaginal smears were recorded every day from the first day of pairing until effective mating to determine the stage of oestrous cycle, and to detect potential adverse effects on the normal cycle.F0 GENERATION:SACRIFICE- females of subgroup C were killed by carbon dioxide inhalation, after successful mating for males, on Gestation Day twenty for females of subgroup C. These animals were eliminated without necropsy. - untreated F0 males of subgroups A and C were killed by carbon dioxide inhalation after successful mating or at the end of the pairing period and eliminated without necropsy.- F0 animals of subgroup A with a sperm positive vaginal smear were killed by ether inhalation on Gestation Day twenty. Detailed autopsy including an examination of their uterine contents was performed. Those from the same subgroup with a sperm negative vaginal smear at the end of the pairing period were killed by ether inhalation ten days after the last mating day for verification of pregnancy and detailed autopsy.GROSS NECROPSY (TREATED ANIMALS)- a detailed autopsy was carried out on each treated F0 female, except those intended exclusively for toxicokinetic investigations. Organs with macroscopic anomalies were sampled and preserved for a possible histopathological evaluation. Corresponding organs of sufficient controls were preserved for comparison.- the ovaries and oviducts were sampled for a possible light microscopic examination from F0 females of subgroup A which were apparently not pregnant at terminal sacrifice. These organs were discarded if any implantation site was detected after uterus staining.HISTOPATHOLOGY / ORGAN WEIGHTS- no histopathological examination was carried out on organs with macroscopic anomalies, which were sampled from F0 animals at terminal sacrifice.Microscopic examination was performed on:- testes and epididymides of high dose and control F0 males- complete genital tract of any F0 males which did not fertilize their corresponding females- ovaries and oviducts of F0 females which were not pregnant (no implantation site detected after uterus staining)The other organs, including the macroscopic anomalies, were kept in fixative without any further investigation.
Ovaries and uterine content:
EXAMINATION OF UTERINE CONTENTS (F0 FEMALES OF SUBGROUP A)- after hysterectomy, the corpora lutea, the implantation sites, the live foetuses and the resorptions were counted- an external macroscopic examination was performed on the foetuses and the placentas, which were then weighed individually- the sex of each foetus was determined either during the organ inspection or during the evisceration of those intended for the skeletal examination
Fetal examinations:
Examination of foetuses of F0 females of subgroup A)Foetuses were killed and half of the foetuses from each litter were allocated to skeletal examination, and half of them were allocated to soft tissue inspection- skeletal examination was performed on foetuses of all groups- organ examination included only foetuses of the high dose group and control group because no biologically significant difference was observed in the findings between these two groups.- the sex of each foetus was determined either during the organ inspection or during the evisceration of those intended for the skeletal examination.
Statistics:
Please refer to the field "Any other information on materials and methods incl. tables" below.
Indices:
Fertility Index
Details on maternal toxic effects:
Maternal toxic effects:no effectsDetails on maternal toxic effects:F0 females from subgroup A (F0 female general condition and fertility)CLINICAL SIGNS AND MORTALITY- administration of S 12911-2 to F0 females of subgroup A, from fourteen days prior to pairing until Gestation Day seventeen, did not provoke mortality or changes in behaviour, but only discolouration of faeces for all animals of all dose groupsBODY WEIGHT AND FOOD CONSUMPTION- body weight of F0 females was not adversely affected by the test substance in any of the treated groups- treatment of F0 females from subgroup A with S 12911-2 did not interfere with feed consumption up to the highest dose administeredREPRODUCTIVE PERFORMANCE- the number of effectively mated F0 females (sperm positive vaginal smear) was similar for all groups- the effective mating rate of the treated F0 females was not affected by the test substance up to the high dose- the mean time necessary for successful copulation, expressed in days, was similar for all groups- the fertility index was similar for F0 females from treated and control groups- evaluation of the uterine contents in the F0 females on day twenty of gestation revealed a slight and not statistically significant increase in the pre-implantation loss rate at 1000 mg/kg/day (0.24 versus 0.14 in controls). Consequently, the mean numbers of implantation sites and live foetuses were slightly decreased at 1000 mg/kg/day (11.1 versus 12.1 in controls for the former, and 10.1 versus 11.4 in controls for the latter). These differences were also not statistically significant. The post-implantation loss rate was also increased at 1000 mg/kg/day (0.17 against 0.05 in controls, not statistically significant), but without increase in early and late resorptions. All these parameters showed generally better values for females treated at 500 and 750 mg/kg/day than for controls.GROSS PATHOLOGY- autopsy revealed no macroscopic organ anomalyHISTOPATHOLOGY- systematic histopathological examination of the ovaries and the oviducts in the non-pregnant females did not reveal any abnormal findingWATER CONSUMPTIONMean water consumption of F0 females from subgroup A was slightly higher in the treated groups than in the control group during the pre-pairing period. However, there was no dose-response relationship, and the differences between treated and control groups were not statistically significant (26.6, 28.8, 31.3, 28.8 g/day at 0, 500, 750 and 1000 mg/kg/day respectively during the second week). During gestation mean water consumption remained higher in the treated than control groups (35.0, 37.9, 41.0, 40.1 g/day on the average at 0, 500, 750 and 1000 mg/kg/day respectively). The differences in group mean values for the entire gestation period, between treated and control groups, were statistically significant at 750 and 1000 mg/kg/day (p < 0.01 and p< 0.05 respectively).
Dose descriptor:
NOAEL
Remarks:
(F0 female)
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: other:
Dose descriptor:
NOAEL
Effect level:
< 500 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: other:
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effectsDetails on embryotoxic / teratogenic effects:F0 females from subgroup A (embryo-foetal development)External examination:- macroscopic external examination of foetuses revealed no findings related to the test substanceVisceral examination:- soft tissue inspection included only foetuses of females treated at the high does (1000 mg/kg/day) and control foetuses - all findings belonged to the spontaneous anomalies of the strain, and no biologically significant difference was observed in the findings between these two groups. Skeletal examination:- an increase in the frequency of delays in the ossification of two sternebrae at all doses (1.9, 11.9, 19.2 and 13.2% at 0, 500, 750 and 1000 mg/kg/day respectively), and caudal vertebrae at all doses (6.7, 9.2, 14.4 and 8.8% at 0, 500, 750 and 1000 mg/kg/day respectively)- an increase in the frequency of variations, which were characterized by wavy ribs, bent bones (scapula, radius, ulna, femur, tibia, fibula), shortened and thickened, humerus, and misshapen clavicle. the percentage of foetuses and litters affected by wavy ribs in each group was 18.3 and 47.4% for controls, 70.6 and 94.7% at 500 mg/kg7day, 75.0 and 100% at 750 mg/kg/day, and 89.0 and 88.9% at 1000 mg/kg/day- the percentage of foetuses and litters affected by at least one skeletal variation of the limbs (clavicle included) in each group was 1.0 and 5.3% for controls, 21.1 and 42.1% at 500 mg/kg/day, 22.1 and 41.2% at 750 mg/kg/day, and 61.5 and 77.8% at 1000 mg/kg/day- one stunted foetus at 0 and 1000 mg/kg/day- one foetus at 750 mg/kg/day presenting one thoracic vertebra limited to a left hemivertebra and a right vertebral arch, and misshapen vertebrae (one thoracic and one lumber)- one foetus at 1000 mg/kg/day presenting one thoracic vertebra limited to the left transverse process, and two fused ribs.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: teratogenicity
Abnormalities:
not specified
Developmental effects observed:
not specified

TOXICOKINETIC EVALUATION

- after administration of S 12911 -2 to to F0 females from subgroups C, strontium and ranelate were detected in the plasma of all animals. This result proves that the compound was absorbed at each dose level.

- mean accumulation ratios were close to unity, however, it was unclear whether steady state was reached.

- AUC24 -values of strontium in females increased proportionally to dose on MD-1, but significantly less than proportionally to dose on Gestation Day 6. No correlation was found on Gestation Day 17.

- AUC24 -values of ranelate in females increased proportionally to dose on MD-1, Gestation Day 6 and gestation Day 17. No correlation was found on Lactation Day 12.

- Cmax-values of strontium and ranelate increased significantly less than proportionally to dose on MD-1 and proportionally to dose on Gestation Day 6. No correlation was found on Gestation Day 17.

Conclusions:
No observed adverse effect levels (NOAELs) of S 12911-2 in the rat:- The NOAEL for F0 male and female general toxicity and fertility is 1000 mg/kg/day, corresponding on the last day of the pre-pairing period to AUC24-values of 376 and 276 µg.h/mL of strontium, respectively, and 16.5 and 13.8 µg.h/mL of ranelate, respectively.- The NOAEL for embryo-foetal development is less than 500 mg/kg/day, mainly with regard to the skeletal variations, which were however, reversible during postnatal development. The maternal systemic exposure (AUC24) at 500 mg/kg/day corresponded to 192 µg.h/mL of strontium and to 7.5 µg.h/mL of ranelate on Gestation Day 17.- The NOAEL for teratogenicity is 1000 mg/kg/ day, corresponding to a maternal AUC24 value on Gestation Day 17 of 302 µg.h/mL for strontium and of 16.7 µg.h/mL of ranelate.
Endpoint:
developmental toxicity
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
1998-05-01 to 1998-08-26
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study reliable without restrictions
Justification for type of information:
Strontium metal completely dissolves upon contact and during the reaction with water under a strong evolution of gas and an immediate precipitation of a white crystalline solid, presumably strontium hydroxide (Sr(OH)2). The water solubility test of strontium (OECD TG 105) indicates a high dissolution from strontium metal (6.74 g/L at 20°C, determined as dissolved strontium, separated by filtration from undissolved test item and precipitates), a rapid formation of Sr2+ + 2OH- + H2 (g) and a corresponding increasing solution pH to a pH > 13. Strontium ions are highly mobile, occur only in one valence state (2+), i.e. are not oxidized or reduced, and do not form strong complexes with most inorganic and organic ligands (Krupka et al. 1999. EPA 402-R-99-004B; Salminen et al. 2015; Carbonaro and Di Toro. 2007. Geochim Cosmochim Acta 71 3958–3968; Carbonaro et al. 2011. Geochim Cosmochim Acta 75: 2499-2511 and references therein). Thus, it may be assumed that systemic toxicological effects (not local) are related to the strontium ion. Therefore, the assessment of the systemic toxicity of strontium is based on elemental strontium concentrations. Read-across of systemic toxicity data available for soluble strontium substances is applied since the strontium ions determine the toxicological potential of strontium.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
adopted 2001-01-22
Deviations:
yes
Remarks:
- gravid uteri including the cervix were not weighed
Qualifier:
equivalent or similar to
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Version / remarks:
, 2008
Qualifier:
according to
Guideline:
other: ICH Harmonised Tripartite Guideline S5 - Detection of toxicity to reproduction for medical products to male fertility, Washington June 24, 1993; ICH Harmonized Tripartite Guideline, Addendum: Toxicity to male fertility, July 1996.
GLP compliance:
yes
Limit test:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS - New Zealand White rabbits (SPF)- Source: C.P.A., 351, rue Roquemaure, 45160 OLIVET, France- Age on mating day: sixteen to nineteen weeks- Weight on mating day: 2.9 to 3.9 kg- Housing: the F0 females were housed in individual cages; cage measurements: length = 75 cm, depth = 54.5 cm, height = 40 cm- Diet (ad libitum): U.A.R (Usine Alimentation Rationnelle, Epinay-sur-Orge, France) sterilized feed pellets- Water (ad libitum): sterilized drinking waterENVIRONMENTAL CONDITIONS- Temperature: 19 ± 2°C- Relative humidity: 55 ± 15%- Air changes: 14 to 19 times per hour- Photoperiod (hrs dark / hrs light): 12/12- Dust level at filter outlet: less than 40 000 particles ≥ 0.5 µm per m^3 an no particle > 5 µm
Route of administration:
oral: gavage
Vehicle:
other: hydroxyethylcellulose
Details on exposure:
VEHICLE- Batch no.: EI922- Physical state: powder- Stability: until April 2000- Dissolved in demineralized water at the concentration of 1% w/v.PREPARATION OF DOSING SOLUTIONS:S 12911-2 was suspended in the vehicle prepared as mentioned above.The concentrations of the various preparations were calculated to allow administration at a constant dose volume of 10 mL/kg bw.The dose volume, calculated on the basis of 10 mL/kg bw, was administered by gavage to each F0 female after weighing.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Tests of stability and homogeneity, carried out before the start of the study (Ginot, Y.M., 1990)*, showed that preparations of S12911-2 in 1% hydroxyethylcellulose (w/v) were stable for twenty-six days when stored in stoppered bottles at room temperature, at concentrations spanning those administered.Chemical analysis of the preparations administered during the study showed that measured concentrations were close to the intended values.The pH of the test substance preparations was checked and values of the first preparation were found between 7.3 and 7.5.*Reference:- Ginot, Y.M. Technologie Servier. Stability Study no.: PA.R. TOX.G04.R02.12911.01, 1990.Homogeneity Study No.: PA.R. TOX.G02.R02.12911.01, 1990
Details on mating procedure:
Impregnation procedure: female New Zealand white rabbits, aged at least sixteen weeks, were paired at the breeder's facilities on three or four consecutive days during four weeks (one delivery per week to the testing laboratory Biologie Servier). During the first three weeks fifty-five female rabbits were mated on the basis of five females per day. During the fourth week twenty-four females were mated on the basis of six females per day. Each female was paired successively with two different untreated mature male rabbits from the same strain. Effective mating was checked visually and the mating day was considered as day zero of gestation.
Duration of treatment / exposure:
Day six of gestation to day eighteen of gestation inclusive
Frequency of treatment:
Once daily and seven days a week
Duration of test:
ca. 30 days
No. of animals per sex per dose:
Treatment groups: 19 mated female rabbitsControl group: 18 mated female rabbits
Control animals:
yes, concurrent vehicle
Details on study design:
Dose selection rationale:The doses were chosen on the basis of the results from the preliminary reproductive toxicity study with New Zealand White rabbits, in which pregnant females were treated at doses of 100, 300, 1000 or 1500 mg/kg, expressed as anhydrous product, once daily by gavage, from day six to day eighteen of gestation.The results showed that S 12911-2 was well tolerated and did not provoke maternotoxic effects up to the highest dose of 1500 mg/kg. Embryotoxic effects, occurred at 1500 mg/kg and were characterized by an increase in the number of early resorptions and, as a consequence, in the post-implantation loss rate. However, the number of live foetuses was not affected. No drug-related foetotoxic or teratogenic effect was observed, whatever the dose used.Systemic exposure, expressed as AUC24, increased proportionally with dose between 100 and 1500 mg/kg, reaching a maximum value of 984 µg x h/mL at the highest dose. After repeated administration, the compound did not accumulate in animal plasma or very slightly (0.82 to 2.1-fold), except for one female treated at 1000 mg/kg 84 times, 1456 µg x h/mL).The dose of 1500 mg/kg represents the limit of feasibility in terms of homogenous ad stable suspension in relationship with a dose volume tolerable by pregnant does. This dose was used again as highest dose. Two lower doses were determined by a geometric progression using a factor of approximately three. Hence these doses were 150 and 500 mg/kg.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes- Time schedule: during gestation the F0 females were observed at least once daily and during the treatment period clinical monitoring was carried out prior to and after treatment. Any sign of abortion was recorded during gestation. DETAILED CLINICAL OBSERVATIONS: NoBODY WEIGHT: Yes- Time schedule for examinations: each female was weighed daily from the day three of gestation to day thirty of gestation inclusive.FOOD CONSUMPTION AND COMPOUND INTAKE: Yes- Food consumption for each animal determined: Yes- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: NoThe parameter was determined daily from day three to day twenty-nine of gestation inclusive, only for the pregnant females alive on Gestation Day thirty.Mean feed consumption per group, expressed in g/day, were calculated for the following four periods:- from day three to day five of gestation inclusive- from day six to day twelve of gestation inclusive- from day thirteen to day eighteen of gestation inclusive- from day nineteen to day twenty-nine of gestation inclusiveWATER CONSUMPTION AND COMPOUND INTAKE: Yes- Time schedule for examinations: the parameter was determined daily from day three to day twenty-nine of gestation inclusive, only for the pregnant females alive on Gestation Day thirty.Mean water consumption per group, expressed in g/day, were calculated for the following four periods:- from day three to day five of gestation inclusive- from day six to day twelve of gestation inclusive- from day thirteen to day eighteen of gestation inclusive- from day nineteen to day twenty-nine of gestation inclusivePOST-MORTEM EXAMINATIONS: Yes- Sacrifice on gestation day 30- Organs examined: uterine contents, as well as a detailed autopsy on each F0 female.- Organs with macroscopic anomalies were sampled and preserved for a possible histopathological evaluation. Corresponding organs of sufficient controls were preserved for comparison.
Ovaries and uterine content:
The uterine content was examined after termination: YesExaminations included:- Gravid uterus weight: No- Number of corpora lutea: Yes- Number of implantations: Yes- Number of resorptions: Yes- Number of foetuses: YesAn external macroscopic examination was performed on the placentas which were then weighed individually.
Fetal examinations:
- External examinations: Yes, an external macroscopic examination was performed on the foetuses which were then weighed individually.- Soft tissue examinations: Yes- Skeletal examinations: Yes- Head examinations: Yes- The sex of each foetus was determined during organ inspection
Statistics:
1) F0 females during gestation- one way analysis of variance (group): global body weight gain between gestation days 6 and 19- two-way analysis of variance (group, time) with repeated measures on time for: body weight during gestation, every day between days 3 and 5, 6 and 19, 20 and 30; daily mean body weight gain during gestation between days 3, 6, 9, 12, 15, 18, 21, 24, 27 and 30; daily mean feed and water consumption during different periods ( gestation days 3 - 5, 6 - 12, 13 - 18, 19 - 29)2) Implantation - Embryonic and foetal development- Kruskal-Wallis test on the following data: number of corpora lutea, number of implantation sites, number of live foetuses, number of dead foetuses, number of early resorptions, number of late resorptions, loss rate before implantation and loss rate after implantationFor each analysis the critical significance level is 5%.References: - Winer, B.J. Statistical Principles in Experimental Design. Mc Graw-Hill - New York, 160 -167, 518 -539, 1971. Second Edition.- Miller, G. Simultaneous Statistical Inference. Springer-Verlag, 76 -81, 1981.- Conover, J. Practical Nonparametric Statistics, Wiley, 229 -237, 1980.- Snedecor, G.W. & Cochran, W.G. In: Statistical Methods. Iowa State University Press, IA 7th Edition, 250 -251, 1980.- SAS/STAT User's Guide. Version 6, fourth edition, 1990.- IMSL STAT/LIBRARY, 1989.
Details on maternal toxic effects:
Maternal toxic effects:yes. Remark: Effects on body weight and food consumption.Details on maternal toxic effects:- Mortality and clinical signs: administration of S 12911-2 to pregnant female New Zealand White rabbits did not provoke test substance-related mortality or any changes in behaviour.- Body weight: global mean body weight gain of F0 females during treatment, from Gestation Day 6 through Gestation Day 19, was reduced at 1500 mg/kg when compared with that of control females (+270, +249, +257 and +164 g at 0, 150, 500 and 1500 mg/kg respectively). The difference from the control group was statistically significant at 1500 mg/kg (p < 0.01). After the withdrawal of treatment on Gestation Day 19, the mean body weight gain until terminal sacrifice on Gestation Day 30 was similar for all groups (+253, +238, +237 and +241 g at 0, 150, 500 and 1500 mg/kg respectively).- Feed consumption: mean feed consumption of females treated at 150 and 500 mg/kg, was similar to that of controls during the entire treatment period, whereas that of females treated at 1500 mg/kg was lower than that of controls during the first and second week of treatment (150, 154, 158, and 122 g/day at 0, 150, 500, and 1500 mg/kg respectively for the second week). The difference from the control group were statistically significant at 1500 mg/kg (p < 0.01). After the withdrawal of treatment, mean feed consumption showed a slight rebound effect at 1500 mg/kg and was similar for all groups (147, 156, 155 and 163 g/day at 0, 150, 500 and 1500 mg/kg respectively).- Water consumption: mean water consumption of F0 females was similar for all groups during the whole gestation period. The differences between groups were not statistically significant.- Macroscopic pathology: one control female found dead on gestation Day 13, had blood in the thoracic cavity and congestive left pulmonary lobe. One female of the 1500 mg/kg dose level group, found dead on Gestation Day 16, had perforation of the oesophagus and haemorrhagic lungs. Another female of the 1500 mg/kg dose level group, found dead on Gestation Day 15, showed mainly yellowish contents (probably test compound) in the thoracic cavity and partially haemorrhagic left pulmonary lobe. An intubation error is likely to be the origin of these three deaths.One female of the 150 mg/kg dose level group killed prematurely on Gestation Day 4 in the lowest dose group had a sacro-lumbar desmorrhexia and also several localized losses of substance on the mucosa of the stomach fundus.One aborted female of the 500 mg/kg dose level group, showed aerocoly, as well as dark spots on the lungs and white areas on the liver at autopsy.Autopsy at termianl sacrifice on Gestation Day 30 of all remaining F0 females revealed congestive ovarian follicles in one non-pregnant female of the 500 mg/kg dose group, accentuated hepatic lobular pattern in one pregnant female of the same dose group, one localized loss of substance on the fundic mucosa in another pregnant female of the same dose group, and in two pregnant females of the 1500 mg/kg dose group one depressed, greyish area or congestive areas on the fundic mucosa respectively. As these macroscopic findings can be seen spontaneously, they were not attributed to an adverse effect of the test substance, and therefore not subjected to a histopathological examination.- Uterine content: treatment of pregnant females during organogenesis with S 12911-2 up to the top dose of 1500 mg/kg, had no impact on the uterine contents. - Mean foetal and placental weights and sex ratio were also similar between all group.
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes. Remark: Retarded ossification and skeletal variationsDetails on embryotoxic / teratogenic effects:Uterine Content:- no impact on uterine contents, mean foetal and placental weights an sex ratio- a non dose- and treatment-related higher mean rate of pre-implanatation loss in all treated groups (0.13, 0.22, 0.25, 0.18 at 0, 150, 500 and 1500 mg/kg, respectively) resulting slightly lower mean no. of impalntations and mean no. of live foetusesExternal examinations:- articular stiffness in 0.6 , 0.7 2.4 and 6.2% of foetuses at 0, 150, 500 and 1500 mg/kg, respectively (spontaneous finding, not test item-related)- one live foetus of the 500 mg/kg dose group was small sized, with shortened limbs, head deformities and a protruding tongue. This foetus was examined for the skeleton by a double staining method instead of X-ray radiography used for all other live foetuses.Visceral examination:- shortened innominate artery in 5.2, 5.2, 8.1 and 13.8% of foetuses at 0, 150, 500 and 1500 mg/kg, respectively (historical control mean and maximum 12.7 and 37.3%, respectively)- absence of truncus brachiocephalicus in 10.3, 16.4, 16.1 and 17.2% of feotuses at 0, 150, 500 and 1500 mg/kg, respectively (historical control mean and maximum 16.4 and 44.4%, respectively)Skeletal examination:An increase in the frequency of delays in the ossification of several bones, mainly at 1500 mg/kg:- skeletal anaomalies >/= 3 (retarded ossification/live foetuses): 3/17, 2/16, 3/15, 10/18 at 0, 150, 500 and 1500 mg/kg, respectively- hyoid bone (8.3, 9.7, 8.9 and 22.8% at 0, 150, 500 and 1500 mg/kg respectively, historical control mean and maximum 6.4 and 17.2%, respectively)- pubic bone (1.9, 0.0, 1.6 and 5.5% at 0, 150, 500, and 1500 mg/kg respectively, historical control mean and maximum 3.8 and 15.8%, respectively)- talus (3.8, 3.0, 3.2 and 7.6% at 0, 150, 500, and 1500 mg/kg respectively, historical control mean and maximum 7.2 and 23.2%, respectively)- phalanges (0.0, 0.0, 12.1 and 18.6% at 0, 150, 500 and 1500 mg/kg respectively, historical control mean and maximum 5.5 and 15.8%, respectively)An increase in the incidence of skeletal variants, mainly at 1500 mg/kg:- skeletal variations >/= 3 (live and dead foetuses): 6/17, 6/16, 7/15, 11/18 at 0, 150, 500 and 1500 mg/kg, respectively- frequency of supernumerary ribs, mainly from 500 mg/kg (34.6, 36.6, 47.6 and 57.2% at 0, 150, 500, and 1500 mg/kg respectively, historical control mean and maximum 22.1 and 45.5% respectively)- wavy ribs only at 1500 mg/kg (8.3%, unknown in this rabbit strain)- two foetuses (1.4%) with one or two bent femurs at 1500 mg/kg (unknown in this rabbit strain)- five cases of malformed foetuses: two in the control , one at 500 mg/kg and two at 1500 mg/kg
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: fetotoxicity
Dose descriptor:
NOAEL
Effect level:
1 500 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: embryotoxicity
Dose descriptor:
NOAEL
Effect level:
1 500 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: teratogenicity
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
S 12911-2 did not provoke any teratogenic effect.Under the conditions of this study, the NOAELs of S 12911-2 were 500 mg/kg for materno- and foetotoxicity, and 1500 mg/kg for embryotoxicity and teratogenicity.
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
172 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The study to examine the effects of strontium ranelate on embryo-fetal and pre-/postnatal development (K. Momburg, 2001) of rats is regarded as relevant and reliable to evaluate the effects of strontium; i.e. a reliable GLP reproduction toxicity study with strontium ranelate (Oral reproduction toxicity study in the Wistar rat (male and female fertility/embryo-fetal and postnatal development) addressing embryo-fetal and pre-/postnatal development of offspring according to the ICH guideline on Detection of Toxicity to Reproduction of Medicinal Products, June 24, 1993, and with the ICH Guideline Addendum: Toxicity to male fertility, July 1996. In addition, a study on the embryo-fetal development of rabbits (K. Momberg 1999) with strontium ranelate is available to complete the database for this endpoint.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the overall evaluation of the available data for soluble strontium substances on reproduction and developmental toxicity, a classification and labelling for reproduction of strontium metal is not considered to be justified according to Regulation (EC) 1272/2008.