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Administrative data

Endpoint:
biochemical or cellular interactions
Type of information:
experimental study
Adequacy of study:
supporting 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:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: Please see below
Principles of method if other than guideline:
Several different in vitro Pharmacology studies were performed according to methods described in the following publications:

Test 1: Binding Assay according to Pristupa, Z.B. et al. (1994), Mol. Pharmacol., 45 : 125-135.
Test 2: Cellular and Nuclear Receptor Functional Assay according to Zhou, Q.Y. et al. (1990), Nature, 347 : 76-80.
Test 3 and 4: Tissue Bioassay according to Zanzottera, D. et al. (1998), Brit. J. Pharmacol., 123 : 730-736 and Steins and, O.S. and Hieble, J.P. (1978), Science, 199 : 443-445.
Test 5: Enzyme and Uptake Assay according to Janowsky, A. et al. (1986), J. Neurochem., 46 : 1272-1276.
GLP compliance:
no
Type of method:
in vitro
Endpoint addressed:
not applicable

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Batch No.of test material: L82-66

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Final dilution of a stock liquid: 1.E-02 M DMSO

FORM AS APPLIED IN THE TEST (if different from that of starting material)
Liquid

Administration / exposure

Vehicle:
DMSO
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
Test 1: 120 min
Test 2: 30 min
Test 3 and 4: no treatment duration
Test 5: 15 min
Doses / concentrations
Dose / conc.:
0 other: M
Remarks:
All assays were performed with this concentration.

Results and discussion

Any other information on results incl. tables

Test 1 Binding assay

Test concentration (M)

% Inhibition of Control Specific Binding

1st

2nd

Mean

1.0E-05

0.9

7.4

4.1

 

Reference compound results

Compound

IC50 (M)

Ki(M)

nH

BTCP

1.3E-08

6.7E-09

1.3

 

Test 2 Cellular and Nuclear Receptor Functional Assay

Agonist effect

Test concentration (M)

% Inhibition of Control Agonist response

1st

2nd

Mean

1.0E-05

0.2

0.3

0.3

 

Reference compound results

Compound

EC50 (M)

nH

Dopamine

4.4E-08

n/a

 

Antagonist effect

Test concentration (M)

% Inhibition of Control Antagonist response

1st

2nd

Mean

1.0E-05

-2.1

-14.7

-8.4

 

Reference compound results

Compound

IC50 (M)

KB (M)

nH

SCH 23390

4.0E-09

6.4E-10

n/a

 

Test 3 and 4: Tissue Bioassay

D1 receptor

In the rabbit splenic artery pre-contracted with U-46619, the D1 receptor agonist SKF 82958 induced a concentration-dependent relaxation which was inhibited by the antagonist R(+)SCH 23390 in a concentration-dependent manner.

The test substance did not affect twitch contraction amplitude at the tested concentration.

In the tissues previously exposed to SKF 82958, the test substance did not significantly reverse the SKF 82958 induced decrease in the twitch contraction amplitude.

These results indicate that test compound does not have an activity at D1 receptors in this tissue.

Agonist effect

Compound

Test concentration (M)

% of control SKF 82958 Response

1st

2nd

Mean

Test substance

1.0E-07

0

0

0

1.0E-06

0

0

0

1.0E-05

0

-3

2

SKF 82958

1.0E-07

13

14

14

3.0E-07

69

64

67

1.0E-06

106

98

102

+ R(+)SCH 23390

1.0E-05

-3

-2

-2

 

Antagonist effect

Compound

Test concentration (M)

% of control SKF 82958 Response

1st

2nd

Mean

Test substance

1.0E-07

100

100

100

1.0E-06

101

104

103

1.0E-05

102

104

103

R(+)SCH 23390

1.0E-06

84

88

86

3.0E-06

50

45

48

1.0E-05

-4

-6

-5

 

The results are expressed as a percent of the control response to SKF 82958 (relaxation) (mean values; n=2)

D2 receptor

In the field-stimulated rabbit ear artery, the D2 receptor agonist (-)quinpirole induced a concentration-dependent decrease in the twitch contraction amplitude which was inhibited by the antagonist (-)sulpiride.

The test substance did not affect twitch contraction amplitude at the tested concentrations.

In the tissues previously exposed to (-)quinpirole, the test substance did not significantly reverse the (-)quinpirole-induced decrease in the twitch contraction amplitude.

These results indicate that the test substance does not have an activity at the D2 receptor in this tissue.

Agonist effect

Compound

Test concentration (M)

% of control (-) Quinpirole Response

1st

2nd

Mean

Test substance

1.0E-07

0

0

0

1.0E-06

-3

-1

-2

1.0E-05

-12

-11

-12

(-)Quinpirole

3.0E-08

12

24

18

1.0E-07

58

69

64

3.0E-07

101

99

100

+ (-)Sulpiride

3.0E-06

16

20

18

 

Antagonist effect

Compound

Test concentration (M)

% of control (-)Quinpirole Response

1st

2nd

Mean

Test substance

1.0E-07

97

97

97

1.0E-06

90

82

8

1.0E-05

82

74

78

(-)Sulpiride

3.0E-08

85

85

85

3.0E-07

42

51

47

3.0E-06

1

-1

0

The results are expressed as a percent of the control response to (-)Quinpirole (decrease in twitch contraction amplitude) (mean values; n=2)

Test 5: Enzyme and Uptake assay

Test concentration

% Inhibition of Control Values

1st

2nd

Mean

1.0E-05 M

-3.2

-2.2

-2.7

 

Reference compound results

Compound

IC50 (M)

nH

GBR 12909

2.2E-09

n/a

Applicant's summary and conclusion