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Toxicological information

Repeated dose toxicity: other routes

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Administrative data

Endpoint:
short-term repeated dose toxicity: other route
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1982
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: no GLP applied, no regulatory guidelines cited, but extensive individual data reported which allow good reconstruction of the summary data
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1982
Report Date:
1982

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
Intravenous injection of alpha-TGIC into the tail vein of mice
GLP compliance:
no
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
alpha-TGIC, purified, batch no. 71689 NF

Test animals

Species:
mouse
Strain:
CD-1
Sex:
male/female
Details on test animals and environmental conditions:
275 CDF-1 mice were acclimatized for 7 days, and maintained in plastic cages in groups of 5 males or females, at 21 +/- 1°C, 50 +/- 10% relative humidity, and a 12-hour dark/light cycle.Food and water was provided ad libitum

Administration / exposure

Route of administration:
intravenous
Vehicle:
water
Details on exposure:
alpha-TGIC was dissolved in water containing 2% mannose.
Injections were made into the tail vein within 1 - 2 seconds at a volume of 25 ml/kg bw
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
(A) single injections with 29 days observation period (some 33 days)
(B) 5 daily injections (tail vein) with 33 days observation period
Frequency of treatment:
(A) single dose
(B) 5 daily doses
Doses / concentrations
Remarks:
Doses / Concentrations:
(A) Dose levels of 50, 74.8, 111.8, 167.2, and 250 mg/kg were applied in a range-finding study
for the main LD50 study were 116.4, 131.3, 148.2, 167.2, 188.6, 121.8, and 240.1 mg/kg
(B) doses of 41.5, 46.5, 52.1, 58.3, 65.3, 73.1, and 81.9 mg/kh bw.
doses of 0, 148.66 mg/kg (LD50), 120.66 mg/kg (LD10, and 60.33 mg/kg (LD5),
No. of animals per sex per dose:
(A) groups of 10 male and 10 female mice
(B) groups of 10 mice (5 males & 5 females)
Control animals:
yes, concurrent vehicle
Details on study design:
Single injections of TGIC in mannitol were made in to the tail vein within 1-2 seconds. Dose levels of 50, 74.8, 111.8, 167.2, and 250 mg/kg were applied in a range-finding study, with an injection volume of 25 ml/kg bw. Animals were observed for 29 days. Single injections of TGIC in mannitol for the main LD50 study were 116.4, 131.3, 148.2, 167.2, 188.6, 121.8, and 240.1 mg/kg with groups of 10 male and 10 female mice. Individual bw were taken on days 1, 8, 15, 22, and 29 post injection. Lethality study with 5 daily injection doses was performed as the single injection experiment with groups of 15 male and 15 female mice and doses of 41.5, 46.5, 52.1, 58.3, 65.3, 73.1, and 81.9 mg/kg bw. Individual body weight were taken on days 1, 2, 3, 4, 5, 12, 19, 26, 33. A single dose toxicity study was performed with 5 groups of 10 mice (5 males & 5 females) with doses of 0, 148.66 mg/kg (LD50), 120.66 mg/kg (LD10, and 60.33 mg/kg (LD5), and observations were made twice per day for the 1st week, and daily thereafter until sacrifice on day 29; body weight was measured on day 1, 2, 7, 10, 14, and 21; all animals dosed with LD50 and half of the other dose groups were sacrificed on day 4, the remaining ones on day 29. Haematology, blood chemistry, and histopathological examinations of major organs were made. In a five day toxicity study, the same procedure as for the single dose toxicity study was applied , and the dose levels were 0, 26.62, 53.24, and 61.55 mg/kg bw. Animals were sacrificed on day 8 and 33.

Examinations

Observations and examinations performed and frequency:
Individual bw were taken on days 1, 8, 15, 22, and 29 post injection. Lethality study with 5 daily injection doses was performed as the single injection experiment with groups of 15 male and 15 female mice and doses of 41.5, 46.5, 52.1, 58.3, 65.3, 73.1, and 81.9 mg/kg bw. Individual body weight were taken on days 1, 2, 3, 4, 5, 12, 19, 26, 33.
observations were made twice per day for the 1st week, and daily thereafter until sacrifice on day 29; body weight was measured on day 1, 2, 7, 10, 14, and 21; all animals dosed with LD50 and half of the other dose groups were sacrificed on day 4, the remaining ones on day 29. Haematology, blood chemistry,
Sacrifice and pathology:
histopathological examinations of major organs were made.
Other examinations:
none
Statistics:
none

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
not examined
Details on results:
In the RF-study the range of the LD50 was 111.8 – 250 mg/kg (all death occurred during days 6-8, and only in groups 167.2 (5710) and 2500 mg/kg (10/10). Clinical signs were diarrhea and rough hair. The intravenous LD50 was 152.4 mg/kg (males) and 145.1 mg/kg (females), and 148.7 mg/kg (combined), and deaths occurred between days 5 – 8. In the surviving animals the clinical signs of toxicity disapeared until day 12. Slight reduction of body weights was observed in all dose groups on day 8 and lasted until day 15. In the 5-day dosing lethality study the intravenous LD50 was 59.12 mg/kg (males) and 64.22 mg/kg (females) and 61.55 mg/kg (combined), deaths occurred during days 6 – 14 (with a single exception on day 19).Toxic sings were rough hair, depression, diarrhea, hunched posture, discolored, necrotic and blistered tails, and excitability. On day 12 all dosed groups showeed a significant body weight reduction in a dose dependent manner, but these revocered towards the end of the study (day 33). In the single dose toxicity study in mice (intravenous injection), no animals died, and no clinical signs were recorded until day 4, but rough hair coat was obserd on days 5-8 in all animals dosed with LD10, but not al the lower dose. Body weigth loss occurred only at the presumed LD50 until day (terminal sacrifice of those animals), but not in the lower dose. Reduction of reticulocytes in all dosed animals and of white blood cells in the high doses group with complete recovery by day 29 of RBCs, but not WBCs was observed. Mean absolute and relative organ weights (liver, kidney heart) were reduced on day 4 at the high and intermediate dose group (males and females), but recovered by day 29 (end of study). Microscopically, drug-related lesions were haematopoietic hypoplasia of the bone-marrow, lymphoid depletion of the thymus, spleen and lymph nodes, and atrophy of the ovaries in the two dosed groups at day 4 but not at day 29 where full recovery was observed. In the toxicity study with 5 consecutive injections of TGIC, toxic signs were recorded from day 5-8, and were diarrhea, rough fur, hunched posture and decrease in water intake. Body weigth reduction was observed in the upper two dose groups between day 6 and 10, and full recovery occurred at day 33 in all surviving animals. On day 8, a decrease of reticulocytes, platelets and white blood cells was observed in all dose groups, and recovery was almost complete at day 33. Mean absolute and relative organ weight changes were observed in the upper two dose groups at day 8 in liver , kidney and heart, but no difference was observed at day 33 in all dose groups. Drug-related lesions were only seen at day 8 (but not day 33) in all TGIC treated groups, and were lymphoid depletion of the thymus, spleen and lymph nodes, haematopoietic hypoplasia of the bone-marrow and necrosis of intestinal mucosa.

Effect levels

Dose descriptor:
LOAEC
Effect level:
> 20 - <= 25 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: mortality, haematological effects, body weight reduction

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The intravenous single dose combined LD10, LD50 and LD90 were 120.7, 148.7 and 183.2 mg/kg bw (in mice). A dose-response relationship was observed with respect to toxic signs, and toxicity was expressed only 2-4 days post application, and were reversible after a recovery period. The main effects were on the haematopoietic system of mice, with depletion of reticulocytes lymphocytes and white blood cells and platelets. Mainly affected were bone-marrow, lymph nodes, thymus and spleen. Atrophy of the ovary was observed after a single dose in some females.
Executive summary:

The intravenous single dose combined LD10, LD50 and LD90 were 120.7, 148.7 and 183.2 mg/kg bw (in mice). A dose-response relationship was observed with respect to toxic signs, and toxicity was expressed only 2-4 days post application, and were reversible after a recovery period. The main effects were on the haematopoietic system of mice, with depletion of reticulocytes lymphocytes and white blood cells and platelets. Mainly affected were bone-marrow, lymph nodes, thymus and spleen. Atrophy of the ovary was observed after a single dose in some females.