disodium 4,4'-bis[(prop-2-enoyl)amino][1,1'-biphenyl]-2,2'-disulfonate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 - 20 Feb 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

Adopted in 2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Slovak National Accreditation Service, Bratislava, Slovak Republic

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

disodium 4-[(1-hydroxyprop-2-en-1-yl)amino]-4'-(prop-2-enamido)-[1,1'-biphenyl]-2,2'-disulfonate

EC Number:

812-549-0

Cas Number:

881539-89-5

Molecular formula:

C18H14N2Na2O8S2

IUPAC Name:

disodium 4-[(1-hydroxyprop-2-en-1-yl)amino]-4'-(prop-2-enamido)-[1,1'-biphenyl]-2,2'-disulfonate

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: VELAZ PRAHA, Czech Republic
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: 184 - 211 g (range)
- Fasting period before study: Animals were fasted 10 - 12 hours prior to dosing (food but not water was withheld over night). Following the period of fasting, animals were weighed, and the test item administered. After test item administration, food was withheld for a further 3 - 4 h.
- Housing: Three animals per cage were housed in plastic cages suspended on stainless steel racks. Bedding used: SAFE 3/4 S, JRS J. Rettenmaier & Söhne GmbH + Co KG.
- Diet: The laboratory food ssniff (ssniff Spezialdiäten GmbH, Germany) was available ad libitum.
- Water: The animals received tap water for human consumption ad libitum. The quality of drinking water is periodically analyzed and recorded.
- Acclimation period: The animals were acclimated under the conditions identical to the conditions during the experiment 5 days prior to the start of treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 50 - 60
- Air changes: Animal room was equipped with central air conditioning.
- Photoperiod (hrs dark / hrs light): 12 /12

IN-LIFE DATES: From: 04 Feb 2020 To: 20 Feb 2020

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

water for injection

Details on oral exposure:

VEHICLE
- Amount of vehicle (gavage): 5 mL/kg bw
- Justification for choice of vehicle: An aqueous solution/suspension/emulsion is a common vehicle in toxicity studies like OECD TG 423.
- Lot/batch no.: 18H0902 (Bieffe Medital S. p. A, Italy)

MAXIMUM DOSE VOLUME APPLIED
Administration volume was 5 mL/kg bw.

DOSAGE PREPARATION
The required amount of the test item was mixed with vehicle shortly before administration.

CLASS METHOD
- Rationale for the selection of the starting dose: A limit dose of 2000 mg/kg bw was used as a starting dose because available information indicated that the test item was likely to be non-toxic regarding acute toxicity.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6 females

Control animals:

no

Remarks:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for clinical signs individually immediately after administration of the test item and 0.5, 1, 2, and 4 h later, and daily thereafter for the next 14 days. Individual body weights of animals were measured immediately prior to administration of the test item and weekly thereafter.
- Necropsy of survivors performed: Yes.
- Examinations performed: All test animals were subjected to gross necropsy.

Statistics:

Not applicable.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study period (see Table 1 under "Any other information on results incl. tables").

Clinical signs:

other: No clinical signs of toxicity were observed up to the end of the 14-day observation period.

Gross pathology:

Necropsy and histopathological examination revealed no substance-related findings.

Any other information on results incl. tables

Table 1. Acute oral toxicity 

Dose [mg/kg bw]	Mortality	Clinical signs
	N*	N*
Females
2000	0/6	0/6

*N= Number of animals/ number of animals used

Table 2: Body weight

Sex	Dose	Animal ID	Body weight (g)	Body weight (g)	Body weight (g)	Body weight difference (g)	Body weight difference (g)	Body weight difference (g)
			Initial	Week 1	Week 2	Week 1 - Initial	Week 2 - Initial	Week 2 - Week 1
female	2000 mg/kg bw	1	211	223	242	12	31	19
female	2000 mg/kg bw	2	198	218	234	20	36	16
female	2000 mg/kg bw	3	195	204	235	9	40	31
female	2000 mg/kg bw	4	185	217	228	32	43	11
female	2000 mg/kg bw	5	189	236	256	47	67	20
female	2000 mg/kg bw	6	184	209	225	25	41	16

Applicant's summary and conclusion

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.

Conclusions:

The test substance AF-959 administered once to 6 female Wistar rats at a limit dose of 2000 mg/kg bw did not cause death, signs of toxicity, body weight losses, or macroscopic pathological changes.