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Toxicity to reproduction: other studies

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Administrative data

Endpoint:
toxicity to reproduction: other studies
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable well documented publication which meets basic scientific principles

Data source

Reference
Reference Type:
publication
Title:
Reproductive Toxic Effects of Cisplatin and Its Modulation by the Antioxidant Sodium 2-Mercaptoethanesulfonate (Mesna) in Female Rats
Author:
Yeh J., Beom Su Kim, Peresie, J.
Year:
2011
Bibliographic source:
Reproductive Biology Insights 2011:5 17–27

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Adult female rats were injected with saline, cisplatin alone, mesna alone (200 mg/kg bw), mesna + saline or mesna + cisplatin, mated with males. The animals were given two i.p. injections one week apart, (mesna 30 minute pretreatment with saline or cisplatin) followed by mating up to ten days after treatment. The animals were euthanized on gestational day 17 and the number corpora lutea, implantation and resorption sites, viable and non-viable fetuses, fetal weights, and the level of progesterone per corpus luteum were determined.
GLP compliance:
no
Type of method:
other: fertility

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: i.p. infusion
Details on test material:
- Name of test material (as cited in study report): Mesna
- Molecular formula (if other than submission substance): C2H5NaO3S2
- Molecular weight (if other than submission substance): 164.18
- Smiles notation (if other than submission substance): C(S)CCS(=O)(=O)O{-}.[Na]{+}
- InChl (if other than submission substance): 1S/C2H6O3S2.Na/c3-7(4,5)2-1-6;/h6H,1-2H2,(H,3,4,5);/q;+1/p-1
- Substance type: organosulphur compound

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan (Indianapolis, IN)
- Age at study initiation: 65-75 days
- Housing: the females were housed with proven male breeders in ratios of 3 females to one male.
- Acclimation period: one week

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
physiological saline
Details on exposure:
VEHICLE
- Justification for use and choice of vehicle (if other than water): i.p. injections were prepared in saline
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
two weeks
Frequency of treatment:
two injections one week apart
Duration of test:
Two weeks (treatments once a week) following by mating period up to ten days (no treatment) and 17 days of gestation (no treatment).
Doses / concentrations
Remarks:
Doses / Concentrations:
200 mg/kg bw
Basis:
no data
No. of animals per sex per dose:
Not reported
Control animals:
yes, concurrent vehicle
Details on study design:
The female rats used in this study were divided into two groups. Group 1 served as the cisplatin mating group and group 2 served as the mesna + cisplatin mating group. The rats in the group 2 (relevant here) were divided into four sub-groups, the control (saline) sub-group, the mesna + saline control subgroup, mesna + cisplatin treatment sub-group, and cisplatin only treatment sub-group. Cisplatin was given at a dosage of 4.5 mg/kg. Mesna was given as a 200 mg/kg dosage 30 minutes prior to the administration of saline or cisplatin. Each rat also received 10 mL of sterile 0.9% NaCl SQ over multiple injection sites following the administration of cisplatin and as needed throughout the course of the study to prevent dehydration. Animals were monitored and weighed daily.
Statistics:
All data are presented as mean +/− standard error of the mean (SEM). Experiments were performed at least three times and the results are presented as the combined data from all experiments. The students’ t test was used to determine statistical difference between the saline and cisplatin groups from the Experiment 1 mating studies. For the Experiment 2 mesna + cisplatin mating studies, analysis of variation (ANOVA) followed by linear trend contrast was used to determine statistical difference. All statistical calculations were carried out using SPSS for Windows, version 11.0. A P < 0.05 was considered statistically different.

Results and discussion

Effect levels

Dose descriptor:
dose level:
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: The administration of mesna + saline did not affect any of the reproductive outcomes compared to the saline control. Mesna protected against cisplatin induced reproductive toxicity.

Observed effects

Experiment 1: Effect of cisplatin on reproductive performance of females:

Cisplatin caused a decrease in the percentage of females that mated to males but there was no change in the fecundity index in the animals treated with cisplatin alone and in the saline control animals. Cisplatin treated animals had an increase in the number of resorption sites and the presence of a non-viable fetus as well as an increase in the percentage of pre-implantation and post-implantation losses in comparison to the control animals. The total number of resorptions per pregnant animal was increased. There was a significant decrease in the number of viable fetuses but no significant differences were in the fetal weight between the saline controls and the cisplatin treated sub-group. The total number of corpora lutea was similar in animals of both groups. The progesterone level per CL between control and cisplatin sub-groups were not statistically different. This suggests that the fetal loss was not directly associated with corpus luteum progesterone production.

Experiment 2:

For the mesna + cisplatin set of experiments, there was a non-significant change in the percentage of females that mated to a male. Of the females that mated with a male, there was no difference in the fecundity index for all sub-three groups. Mesna protected against the pre-implantation and postimplantation losses of the fetuses induced by cisplatin. There was a statistically significant difference in the number of resorptions seen among the control, mesna + cisplatin, and cisplatin sub-groups. Treatment with mesna leads to an increase in the total number of viable fetuses in the mesna + cisplatin compared to cisplatin. The
weight of the viable fetuses was not statistically different among the three sub-groups. After mesna and cisplatin administration, there was no difference in the total number of corpora lutea found in the saline, mesna + cisplatin and cisplatin sub-groups. The measurement of progesterone revealed that there was no difference in the progesterone level per corpus luteum in any of the three sub-groups.

Applicant's summary and conclusion

Conclusions:
The administration of mesna alone did not affect any of the reproductive indices in rats. Mesna protected against cisplatin induced toxicitxy: mesna administered prior to cisplatin resulted in a decrease in the rate of the pre- and post implantation loss, along with a decrease in the number of resorptions and an increase in the number of live fetuses.
Executive summary:

The influence of Mesna on the cisplatin induced reproductive toxicity was studied in rats (Yeh at el., 2011). Cisplatin, a chemotherapeutic agent, is known to cause damage to ovaries in rats and a variety of adverse effects in foetuses of rats and mice. In humans, cisplatin caused damage to ovaries as well. Therefore, the administration of mesna prior to multiple doses of cisplatin was believed to offer protection from the loss of fertility induced by cisplatin. Adult female Sprague Dawley rats were injected with saline, cisplatin alone (4.5 mg/kg bw), mesna alone (200 mg/kg bw), mesna + saline or mesna + cisplatin, and mated with males. The female rats were divided into two groups. Group 1 served as cisplatin mating group, in which effects of cisplatin on reproductive performance of females were studied and group 2 served as the mesna + cisplatin mating group in which effects of mesna on cisplatin induced reproductive toxicity were studied. Group 1 was sub-divided into two treatment sub-groups, the saline control sub-group and the cisplatin treated sub-group. Group 2 was divided into four sub-groups, the control (saline) sub-group, the mesna + saline control subgroup, mesna + cisplatin treatment sub-group, and cisplatin only treatment sub-group. The animals were given two i.p. injections one week apart, (mesna 30 minute pretreatment with saline or cisplatin) followed by mating up to ten days after treatment. The animals were euthanized on gestational day 17 and the number corpora lutea, implantation and resorptions sites, viable and non-viable fetuses, fetal weights, and the level of progesterone per corpus luteum were determined.

In the cisplatin mating group (group 1), cisplatin caused a decrease in the percentage of females that mated to males but there was no change in the fecundity index in the animals treated with cisplatin alone and in the saline control animals. Cisplatin treated animals had an increase in the number of resorption sites and the presence of a non-viable fetus as well as an increase in the percentage of pre-implantation and post-implantation losses in comparison to the control animals. The total number of resorptions per pregnant animal was increased. There was a significant decrease in the number of viable fetuses but there were no significant differences in the fetal weight between the saline controls and the cisplatin treated sub-group. The total number of corpora lutea was similar in animals of both groups. The progesterone level per CL between control and cisplatin sub-groups were not statistically different. This suggests that the fetal loss was not directly associated with corpus luteum progesterone production.

The administration of mesna + saline did not affect any of the reproductive outcomes compared to the saline control. In the mesna + cisplatin mating group, there was a non-significant change in the percentage of females that mated to a male. Of the females that mated with a male, there was no difference in the fecundity index for all sub-three groups. Mesna protected against the pre-implantation and postimplantation losses of the fetuses induced by cisplatin. There was a statistically significant difference in the number of resorptions seen among the control, mesna + cisplatin, and cisplatin sub-groups. Treatment with mesna leads to an increase in the total number of viable fetuses in the mesna + cisplatin compared to cisplatin. The weight of the viable fetuses was not statistically different among the three sub-groups. After mesna and cisplatin administration, there was no difference in the total number of corpora lutea found in the saline, mesna + cisplatin and cisplatin sub-groups. The measurement of progesterone revealed that there was no difference in the progesterone level per corpus luteum in any of the three sub-groups.

In conclusion, prior exposure to cisplatin caused a significant adverse effects on fertility as evidenced by the decreased implantation due to increased fetal loss. The administration of mesna appeared to temper cisplatin damage by lessening the cisplatin effects on fetal resorption.