[2S-[2α,5α,6β(S*)]]-6-[[[[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonyl]amino]phenylacetyl]amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid

Acute Toxicity: other routes

Administrative data

Endpoint:

acute toxicity: other routes

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1977

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

- Principle of test: method for assessing acute toxicity that involves the identification of a dose level that causes evidence of non-lethal toxicity.
- Short description of test conditions: The test item was administered i.v. to groups of 7 Wistar SPF rats per sex (8 or 16 weeks old) at 12 doses ranging from 1.81 to 3.54 g/kg bw test item.
- Parameters analysed / observed: Mortalities and clinical signs were monitored during 7 days post-dosing after which animals were necropsied and subjected to a gross examination.

GLP compliance:

not specified

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6-[[[[(4-ethyl-2,3-dioxo-1-piperazinyl)carbonyl]amino]phenylacetyl]amino]-3,3-dimethyl-7-oxo-, monosodium salt, [2S-[2α,5α,6β(S*)]]-

EC Number:

261-868-6

EC Name:

4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, 6-[[[[(4-ethyl-2,3-dioxo-1-piperazinyl)carbonyl]amino]phenylacetyl]amino]-3,3-dimethyl-7-oxo-, monosodium salt, [2S-[2α,5α,6β(S*)]]-

Cas Number:

59703-84-3

Molecular formula:

C23H26N5O7S.Na

IUPAC Name:

sodium;(2S,5R,6R)-6-[(2S)-2-[4-ethyl-2,3-dioxopiperazin-1-yl)carbonylamino]-2-phenylacetamido]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate

Test material form:

solid

Specific details on test material used for the study:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: dilution with water.

Test animals

Species:

rat

Strain:

Wistar

Remarks:

SPF

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Nippon CLEA Co., Ltd.
- Age at study initiation: 8 and 16 weeks old. The LD50 were compared in the stages of growth.
- Weight at study initiation: 8 weeks: (8 weeks old: males = 220-290 g, females = 160-195 g), 16 weeks (body weight: males 400 to 450 g).
- Diet: ad libitum
- Water: tap water ad libitum
- Acclimation period: one week.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2ºC
- Humidity (%): 60 ± 5%

Administration / exposure

Route of administration:

intravenous

Vehicle:

water

Doses:

12 doses from 1810 mg/kg bw to 3540 mg/kg-bw

No. of animals per sex per dose:

7

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 7 days.
- Frequency of observations and weighing: daily observations.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, macroscopic observations.

Statistics:

95% confidence limits were calculated by Litchfield-Wilcoxon’s method.

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

2 710 mg/kg bw

Based on:

test mat.

95% CL:

>= 2 360 - <= 3 120

Remarks on result:

other: 8 weeks old

Effect levels 2

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

2 790 mg/kg bw

Based on:

test mat.

95% CL:

>= 2 210 - <= 3 500

Remarks on result:

other: 16 weeks old

Effect levels 3

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

2 260 mg/kg bw

Based on:

test mat.

95% CL:

>= 2 100 - <= 2 430

Remarks on result:

other: 16 weeks old

Mortality:

No mortality was observed.

Clinical signs:

At 2000 mg/kg bw, only a decrease in hyperactive locomotor activity was observed.
At 2660 mg/kg bw, transient effects that included lack of control of limb movement and unresponsiveness to stimulae for about 30 min post-administration. The surviving animals recovered about 2h after administration. 4 males and 3 females in this group showed ankylosing convulsions for 15-30 min, and died.

Gross pathology:

At necropsy of the dead animals of the 2660 mg/kg bw group, pulmonary haemorrhage and mild subdural haemorrhage of the brain were found in most cases. These were not observed at necropsy of the surviving animals of the same group.

Any other information on results incl. tables

Table 1. Death rate in acute toxicity tests of T-1220. Rat, i.v.

Age	Dose (g/kg)	Male									Female
		Time in days								Death rate	Time in days								Death rate
		0	1	2	3	4	5	6	7		0	1	2	3	4	5	6	7
8 w	3.54	7								7/7	7								7/7
	3.22	6	0	0	0	0	0	0	0	6/7	7	0	0	0	0	0	0	0	7/7
	2.93	4	0	0	0	0	0	0	0	4/7	4	0	0	0	0	0	0	0	4/7
	2.66	4	0	0	0	0	0	0	0	4/7	3	0	0	0	0	0	0	0	3/7
	2.42	1	0	0	0	0	0	0	0	1/7	3	0	0	0	0	0	0	0	3/7
	2.20	1	0	0	0	0	0	0	0	1/7	0	0	0	0	0	0	0	0	0/7
	2.00	0	0	0	0	0	0	0	0	0/7	0	0	0	0	0	0	0	0	0/7
16 w	2.66	7	0	0	0	0	0	0	0	7/7
	2.42	5	0	0	0	0	0	0	0	5/7
	2.20	3	0	0	0	0	0	0	0	3/7
	2.00	1	0	0	0	0	0	0	0	1/7
	1.81	0	0	0	0	0	0	0	0	0/7

 

Table 2. Summary of results.  

Animal	Route	Age	LD50 (g/kg)
			Male	Female
Rat	i.v.	8 wk	2.71 (2.36 – 3.12)	2.79 (2.21 – 3.50)
		16 wk	2.26 (2.10 – 2.43)

*(): 95% confidence limits, calculated by Litchfield-Wilcoxon’s method.

Applicant's summary and conclusion

Conclusions:

The intravenous LD50 of the test item in rats was > 2000 mg/kg-bw.

Executive summary:

An acute intravenous toxicity study was conducted in order to determine the toxicological properties of the sodium salt of piperacillin with a method similar to OECD guideline 401 (non-GLP). The test item was administered i.v. to groups of 7 Wistar SPF rats per sex (8 or 16 weeks old) at 12 doses ranging from 1.81 to 3.54 g/kg bw test item. Mortalities and clinical signs were monitored during 7 days post-dosing after which animals were necropsied and subjected to a gross examination. All animals died at the highest dose tested, and toxic effects were observed until 2.66 g/kg bw. At 2.00 g/kg bw no mortalities occured, the clinical signs were limited to mild transient locomotor supression, no gross abnormalities were observed at necropsy. The intravenous LD50 was found to be > 2000 mg/kg bw in rats.