Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
29 August 2017 to 13 September 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
OECD GUIDELINES FOR TESTING OF CHEMICALS (423, adopted at 17th Dec. 2001)
Deviations:
yes
Remarks:
Due to technical reason, relative humidity values (maximum of 80 %) outside the expected range of 30-70 % were recorded during the acclimation period. This deviation has no presumed impact on the outcome or integrity of the study.
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris
Deviations:
yes
Remarks:
Due to technical reason, relative humidity values (maximum of 80 %) outside the expected range of 30-70 % were recorded during the acclimation period. This deviation has no presumed impact on the outcome or integrity of the study.
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
EPA Health Effects Test Guidelines (OPPTS 870.1100), United States, EPA 712-C-98-190 (1998)
Deviations:
yes
Remarks:
Due to technical reason, relative humidity values (maximum of 80 %) outside the expected range of 30-70 % were recorded during the acclimation period. This deviation has no presumed impact on the outcome or integrity of the study.
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
diphenylmethyl (6R,7R)-7-[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-[(triphenylmethoxy)imino]acetamido]-3-(hydroxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate
EC Number:
609-461-8
Cas Number:
376653-36-0
Molecular formula:
C44H36N6O6S2
IUPAC Name:
diphenylmethyl (6R,7R)-7-[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-[(triphenylmethoxy)imino]acetamido]-3-(hydroxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate
Test material form:
solid: particulate/powder
Details on test material:
Name: BAL0001022
CAS number: 376653-36-0
Batch/Lot number: 14024R31D
Appearance: Off-white powder
Purity: 87.7%
Expiry date: 31 January 2020
Storage conditions: Under inert gas, protected from light and humidity (tight closed container), frozen (≤ -15 °C)
Safety precautions: Enhanced safety precautions were applied considering the supplied safety datasheet to assure personnel health and safety.
Specific details on test material used for the study:
No further details specified in the study report.

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
Species and strain: CRL:(WI) Wistar rats
Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld
Hygienic level at arrival: SPF
Hygienic level during the study: Standard housing conditions
Number of animals: 6 animals
Sex: Female, nulliparous and non-pregnant
Age of animals at dosing: Young healthy adult rats, 9 weeks old
Body weight at treatment: 227 – 242 g
Acclimatization period: at least 12 days

Husbandry
Animal health: Only healthy animals were used for the test. The staff Veterinarian certified health status.
Number of animal room: 522/1
Housing: 3 animals / cage
Cage type: Type II polypropylene/polycarbonate
Bedding/Nesting: “Lignocel 3/4-S Hygienic Animal Bedding” and “Arbocel crinklets natural” nest building material produced by J. Rettenmaier & Söhne GmbH & Co.KG (D-73494 Rosenberg, Germany) were available to animals during the study.
Lighting period: 12 hours daily, from 6.00 am to 6.00 pm
Temperature: 22 ± 3 °C
Relative humidity: 33-80%
Ventilation: 15 – 20 air exchanges/hour
Enrichment: Animals were housed by group to allow social interaction and with deep wood sawdust bedding to allow digging and other normal rodent activities.
The room temperature and relative humidity were recorded twice daily during the study.

Food and Water Supply
Animals received ssniff® SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest, Germany (Batch no.: 262 21592, expiry date: 31 January 2018), ad libitum, and tap water from the municipal supply, as for human consumption from 500 ml bottles, ad libitum. The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u.36., Hungary).

Animal Identification
Animals were individually identified using numbers written on the tail with an indelible marker pen. The numbers were given on the basis of Citoxlab Hungary Ltd.' s Master File, for each animal allocated to the treatment groups. The cages were identified by cards, with information about study code, sex, dose group, cage number and individual animal numbers.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Remarks:
PEG 400
Details on oral exposure:
Formulation
The test item was freshly formulated at a concentration of 200 mg/mL in the vehicle, in the Pharmacy of Citoxlab Hungary Ltd. on the day of administration. The formulation container was stirred continuously up to finishing the treatment.

Vehicle Selection
The selection of the vehicle was made during trial formulations with the test item. The final choice of vehicle was approved by the Sponsor.
On the basis of the trial formulations with the test item, the vehicle used was PEG 400.
Vehicle: PEG 400 (Poly (ethylene glycol))
Batch number: BCBS1795V
Expiry date: 31 May 2018
Doses:
A limit of 2000 mg/kg bw dose was selected by the Sponsor in line with the guidelines.
No. of animals per sex per dose:
Initially, three females (assigned to Group 1) were treated at a dose level of 2000 mg/kg bw. As the test item did not cause mortality in this group a second group (Group 2) was treated at the same dose level.
Control animals:
no
Details on study design:
Procedure
A single oral gavage administration (dose administered at 10 mL/kg bw) was followed by a 14-day observation period. The day before treatment, food was removed at the end of the working day and the animals therefore fasted for a period of approximately 17 hours. Water was not withheld during this period. Animals were weighed before treatment. The test item was administered by oral gavage in the morning and food made available 3 hours after the treatment.

OBSERVATIONS
Clinical Observations
Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Individual observations were performed on the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Body Weight Measurement
The body weight was recorded the day before treatment (Day -1), on the day of the treatment (Day 0), Day 7 and Day 14.

NECROPSY
All animals were subjected to a necropsy and a macroscopic examination at the end of the study on Day 14. The animals were exsanguinated after verification of narcosis following an injection of pentobarbital sodium (Release; Lot No.: 106075, Expiry Date: July 2018, Produced by: Wirtschaftsgenossenschaft deutscher Tierärzte eG, Siemensstr. 14, 30827 Garbsen, Germany). After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. All gross macroscopic changes were recorded for each animal.
Statistics:
Not specified

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
BAL0001022 did not cause mortality at a dose level of 2000 mg/kg bw.
Clinical signs:
Treatment with BAL0001022 at the dose level of 2000 mg/kg bw did not cause any adverse clinical signs.
Body weight:
There were no treatment related effects on body weight or body weight gain during the observation period.
One animal given 2000 mg/kg bw (No: 627) showed a reduced bodyweight gain (0.39 %) in the second week of the observation period. This change was considered incidental and not ascribed to treatment.
Gross pathology:
There was no evidence of treatment-related macroscopic changes at necropsy at a dose level of 2000 mg/kg bw.

Any other information on results incl. tables

INDIVIDUAL CLINICAL OBSERVATIONS

DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0                                                                SEX: FEMALE

Cage No.

Animal Number

Observations

Observation days

Frequency

0

1

2

3

4

5

6

7-14

30’

1h

2h

3h

4h

6h

1

624

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

20/20

625

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

20/20

626

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

20/20

2

627

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

20/20

628

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

20/20

629

Symptom free

+

+

+

+

+

+

+

+

+

+

+

+

+

20/20

Remarks:              + = present

                         h = hour (s)          ‘ = minute

                        Frequency of observation = number of occurrence of observations / total number of observations

 

INDIVIDUAL BODY WEIGHT AND BODY WEIGHT GAIN

DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0                              SEX: FEMALE

Cage No.

Animal Number

Body weight (g)

Days

Body Weight Gain (g)

-1

0

7

14

-1-0

0-7

7-14

-1-14

1

624

253

242

274

285

-11

32

11

32

625

246

231

255

267

-15

24

12

21

626

256

239

278

280

-17

39

2

24

2

627

247

237

256

255

-10

19

-1

8

628

238

227

255

272

-11

28

17

34

629

252

236

262

281

-16

26

19

29

Mean:

248.7

235.3

263.3

273.3

-13.3

28.0

10.0

24.7

Standard deviation:

6.4

5.5

10.2

11.1

3.0

6.9

8.0

9.5

 

INDIVIDUAL INTERNAL AND EXTERNAL MACROSCOPIC OBSERVATIONS

DOSE LEVEL: 2000 mg/kg bw, Treatment on Day 0                                                     SEX: FEMALE

Cage No.

Animal Number

Necropsy Date/

Necropsy Day

External Observations

Internal Observations

Organ/Tissue

1

624

12 September 2017

Day 14

No external observations recorded

No internal observations recorded

Not applicable

625

12 September 2017

Day 14

No external observations recorded

No internal observations recorded

Not applicable

626

12 September 2017

Day 14

No external observations recorded

No internal observations recorded

Not applicable

2

627

13 September 2017

Day 14

No external observations recorded

No internal observations recorded

Not applicable

628

13 September 2017

Day 14

No external observations recorded

No internal observations recorded

Not applicable

629

13 September 2017

Day 14

No external observations recorded

No internal observations recorded

Not applicable

 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In the absence of any acute toxicity, BAL0001022 can be ranked as "Category 5" or "Unclassified" for acute oral toxicity according to the GHS criteria. Although this study was not designed to determine the acute oral LD50, under the conditions of this study, it is assumed for BAL0001022 to be above 2000 mg/kg bw in female CRL:(WI) rats.
Executive summary:

This single-dose oral toxicity of BAL0001022 was performed according to the acute toxic class method (OECD 423 and Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1. tris) in CRL:(WI) rats.

 

Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. The test item was administered formulated in Poly (ethylene glycol) 400 (PEG 400) at a concentration of 200 mg/mL at a dosing volume of 10 mL/kg bw. A single oral treatment was carried out by gavage for each animal after food had been withdrawn overnight. Food was made available again 3 hours after the treatment. Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. Body weight was measured on Days -1, 0 and 7 and Day 14. All animals were subjected to a necropsy and a macroscopic examination.

 

As no mortality was observed in group 1 a second group (Group 2) of 3 animals was treated at the same dose level using the methods described for group 1.

 

Results

 

Mortality

BAL0001022 did not cause mortality at a dose level of 2000 mg/kg bw.

 

Clinical Observations

Treatment with BAL0001022 at the dose level of 2000 mg/kg bw did not cause any adverse clinical signs.

 

Body Weight and Body Weight Gain

There was no relevant treatment related effects on body weight or body weight gain during the observation period. One animal given 2000 mg/kg bw (No: 627) showed a reduced body weight gain (0.39%) in the second week of the observation period. This change was considered incidental and not ascribed to treatment.

 

Macroscopic Findings

There was no evidence of treatment-related macroscopic changes at necropsy in animals given 2000 mg/kg bw.

 

Conclusion:

In the absence of any acute toxicity, BAL0001022 can be ranked as "Category 5" or "Unclassified" for acute oral toxicity according to the GHS criteria. Although this study was not designed to determine the acute oral LD50, under the conditions of this study, it is assumed for BAL0001022 to be above 2000 mg/kg bw in female CRL:(WI) rats.