2.6.7.8.1. 細菌を用いる復帰突然変異試験
Report Title: Testing for mutagenic activity with Salmonella typhimurium TA1535, TA1537, TA98 and TA100 and Escherichia coli WP2uvrA
Test Article: Levocetirizine No. of Independent Assays: 2 Report No. RRLE99K1101 Strains: Salmonella typhimurium TA98, TA100, TA1535, TA1537,
Escherichia coli WP2uvrA No. of Replicate Cultures: 3 Location in CTD: 4.2.3.3.1.1
Metabolizing System: Aroclor 1254 induced rat liver S9-fraction. No. of Cells Analyzed/Culture: Not applicable GLP Compliance: Yes
Vehicles: For test article and methyl methane sulphonate - Ultra-pure water. For other positive controls - DMSO. Date of Treatment: to 19 Treatment: 2 days (+S9 or -S9)
Cytotoxic Effects: Toxicity at 1000µg/plate (-S9) and from 5000µg/plate (+S9).
Genotoxic Effects: No genotoxicity up to 1000µg/plate (-S9) or up to 5000µg/plate (+S9).
Mean revertant colony counts
TA98 TA100 TA1535 TA1537 WP2uvrA Metabolic
Activation Test Article Conc.
(µg/plate)
1 2 1 2 1 2 1 2 1 2
0 17 20 91 125 11 11 7 8 8 8
31.25 18 19 90 132 6 10 9 6 6 8
62.5 14 19 88 118 10 8 6 8 6 4
125.0 13 18 91 111 8 11 4 9 5 6
250.0 16 15 91 116 8 8 9 5 7 8
500.0 15 15 88 104 8 10 5 10 8 4 Levocetirizine
1000.0 10 21 84 83 8 8 5 6 4 6
2.0 418 241
N-ethyl-N’-nitro-N-nitrosoguanidine
5.0 1951 1451
Methyl methane sulphonate 200.0 453 491
2-nitrofluorene 1.0 616 448
Without activation
9-aminoacridine 80.0 2060 1777
1, 2: Independent experiments; study using batch No. 506, Blank: not tested 2.6.7. 毒性試験概要表
2.6.7 - p. 16
2.6.7.8.1. 細菌を用いる復帰突然変異試験（続き）
Mean revertant colony counts
TA98 TA100 TA1535 TA1537 WP2uvrA Metabolic
Activation Test Article Conc.
(µg/plate)
1 2 1 2 1 2 1 2 1 2
0 22 19 96 139 9 9 11 9 6 6
78.125 31 148 9 14 6
156.25 20 23 102 142 8 7 10 15 5 7 312.5 19 19 94 130 6 9 12 11 6 5 625.0 21 25 94 137 4 12 10 15 8 8
1250.0 21 21 80 106 7 8 10 9 3 6
2500.0 15 23 74 116 8 9 10 7 7 6
Levocetirizine
5000.0 2 60 6 7 2
0.5 347 461 408 490
2.0 196 248 219 187
With activation
2-aminoanthracene
20.0 672 540
1, 2: Independent experiments, Blank: not tested
2.6.7. 毒性試験概要表
2.6.7 - p. 17
2.6.7.8.2. マウスリンフォーマTK試験
Report Title: Mouse lymphoma mutation assay Test Article: Levocetirizine Test for Induction of: Forward mutation from TK+/- to TK-/-. No. of Independent Assays: 2 Report No. RRLE99K1102 Strains: Mouse lymphoma L5178Y (clone 3.7.2 C). No. of Replicate Cultures: 2 (-S9 and +S9) or 4
(solvent control)
Location in CTD: 4.2.3.3.1.2 Metabolizing System: Aroclor 1254 induced rat liver S9-fraction. No. of Cells Analyzed/Culture: Not applicable GLP Compliance: Yes
Vehicles: For test article and methyl methane sulphonate - Sterilised water. For other positive controls - DMSO. Date of Treatment: to 19 Treatment: -S9 1st assay - 4 hours, 2nd assay - 24 hours; +S9 - 4 hours.
Cytotoxic Effects: -S9 from 400µg/mL (4hr exposure), from 300µg/mL (24hr exposure); +S9 from 300µg/mL. (< 10% relative survival or relative suspension growth).
Genotoxic Effects: -S9 - no genotoxicity up to 350µg/mL; +S9 - no genotoxicity up to 300µg/mL.
-S9; Test 1 - 4hr exposure
Day 0 Day 2
Test article Conc. (µg/mL) Cloning efficiency
(%) Relative survival
(%) Cloning efficiency
(%) Mutant fraction
(x106) Ratio small/large colonies
0 102 100 104 88 0.66
50 95 94 NPS NPS -
100 91 89 NPS NPS -
150 103 97 NPS NPS - 200 77 57 119 73 0.76 250 107 70 90 112 0.90 300 97 50 101 74 1.71 350 94 36 92 59 1.04 Levocetirizine
400 58 18 NPT NPT -
Ethyl methane sulphonate 250 83 103 93 551* 0.39
Methyl methane sulphonate 15 54 59 38 1141* 1.69 NPS = Not plated, surplus. NPT = Not plated; toxic
Significantly different from controls (*: p < 0.05). (Statistical analysis by the recommended UKEMS method; study using batch 506.)
2.6.7. 毒性試験概要表
2.6.7 - p. 18
2.6.7.8.2. マウスリンフォーマTK試験（続き）
-S9; Test 2 - 24hr exposure
Day 1 Day 3
Test article Conc. (µg/mL) Cloning efficiency
(%) Relative survival
(%) Cloning efficiency
(%) Mutant fraction
(x106) Ratio small/large colonies
0 108 100 98 62 0.59 50 99 76 117 49 0.66 100 94 54 114 69 0.44 150 119 36 118 70 0.96 200 84 19 111 66 0.63 250 79 8 # # -
300 84 5 NPT NPT -
350 NPT - - - -
Levocetirizine
400 NPT - - - -
Ethyl methane sulphonate 150 59 38 58 1930* 0.45 Methyl methane sulphonate 5 76 52 62 1110* 1.98 NPT = Not plated; toxic. # = The Day 1 RS values for this dose were below the minimum acceptable 10%. Day 3 data were consequently excluded from the assessment.
Significantly different from controls (*: p < 0.05). (Statistical analysis by the recommended UKEMS method.)
2.6.7. 毒性試験概要表
2.6.7 - p. 19
2.6.7.8.2. マウスリンフォーマTK試験（続き）
+S9; Test 1 - 4hr exposure
Day 0 Day 2
Test article Conc. (µg/mL) Cloning efficiency
(%) Relative survival
(%) Cloning efficiency
(%) Mutant fraction
(x106) Ratio small/large colonies
0 96 100 104 84 1.16 12.5 115 106 NPS NPS - 25.0 104 101 NPS NPS - 50.0 88 83 NPS NPS - 100.0 78 54 83 111 0.70
150.0 74 35 92 86 0.91
200.0 62 27 106 131 0.59 250.0 75 22 96 127 0.70 Levocetirizine
300.0 43 11 NPT NPT -
3-methylcholanthrene 2.5 54 55 69 881* 1.04
NPS = Not plated, surplus. NPT = Not plated; toxic.
Significantly different from controls (*: p < 0.05). (Statistical analysis by the recommended UKEMS method).
+S9; Test 2 - 4hr exposure
Day 0 Day 2
Test article Conc. (µg/mL) Cloning efficiency (%)
Relative survival (%)
Cloning efficiency (%)
Mutant fraction (x106)
Ratio small/large colonies
0 105 100 111 65 0.74
50 112 88 NPS NPS -
100 89 64 114 48 0.69 150 69 47 126 59 0.50 200 76 31 96 58 0.58 250 63 27 93 75 1.17 300 52 13 98 77 0.89
350 35 7 NPT NPT -
Levocetirizine
400 17 2 NPT NPT -
3-methylcholanthrene 2.5 81 68 89 805* 0.84
NPS = Not plated, surplus. NPT = Not plated; toxic.
Significantly different from controls (*: p < 0.05). (Statistical analysis by the recommended UKEMS method).
2.6.7. 毒性試験概要表
2.6.7 - p. 20
2.6.7.8.3. ヒト培養リンパ球を用いる染色体異常試験
Report Title: Induction of chromosome aberrations in cultured human peripheral blood lymphocytes. Test Article: Levocetirizine Test for Induction of: Chromosome aberrations No. of Independent Assays: 2 Report No. RRLE06B1736 No. of Replicate Cultures: 4 (solvent), 2 (test article, positive controls) No. of Cells Analyzed/Culture: 100 (except for positive
controls)
Location in CTD: 4.2.3.3.1.3 Metabolizing System: Rat liver (S9) GLP Compliance: Yes Vehicle: Purified water
Treatment: 3+17 hour treatment in the absence and presence of metabolic activation (S9), Experiment 1. 20+0 hour treatment in the absence of metabolic activation (S9) and 3+17 hour treatment in the presence of S9, Experiment 2
Date of Treatments: to 20
Cytotoxic Effects: Experiment 1 - 700 and 650µg/mL in the absence and presence of S9. Experiment 2 - 650 and 700µg/mL in the absence and presence of S9. Higher concentrations exceeded the acceptable toxicity limits for this assay.
Genotoxic Effects: None Experiment 1
Metabolic Activation Test Article Concentration (µg/mL )
Treatment 3+17 hours # Cells with aberrations
including gaps
Treatment 3+17 hours # Cells with aberrations
excluding gaps
Treatment 3+17 hours # Mitotic inhibition† Without Activation Vehicle (Water) 0 1 1 -
Levocetirizine 600.0 2 1 23
650.0 3 3 38
700.0 2 2 44
NQO 2.50 21 20*** -
With Activation Vehicle (Water) 0 3 3 -
Levocetirizine 400.0 2 2 17
550.0 1 1 38
650.0 3 3 47
CPA 12.5 40 39*** -
Study using batch No. 05B201020
# (hours treatment + hours recovery), NQO: 4-Nitroquinoline 1-oxide, CPA: Cyclophosphamide
†: Mitotic inhibition (%) = [1 - (mean MIT/mean MIC)] x 100% (where T = treatment and C = negative control), ***: p≤0.001 2.6.7. 毒性試験概要表
2.6.7 - p. 21
2.6.7.8.3. ヒト培養リンパ球を用いる染色体異常試験（続き）
Experiment 2 Metabolic
Activation Test Article Concentration (µg/mL)
Treatment 3+17 hours # Cells with
aberrations including gaps
Treatment 3+17 hours # Cells with
aberrations excluding gaps
Treatment 3+17 hours # Mitotic
inhibition†
Treatment 20+0 hours # Cells with
aberrations including gaps
Treatment 20+0 hours # Cells with
aberrations excluding gaps
Treatment 20+0 hours # Mitotic
inhibition†
Vehicle (Water) 0 3 1 -
Levocetirizine 300.0 2 0 0††
550.0 3 3 0
600.0 0 0 38
650.0 3 1 65
Without Activation
NQO 5.00 47 40*** -
Vehicle (Water) 0 2 2 -
Levocetirizine 600.0 5 4 2
650.0 4 2 27 700.0 4 2 57 With Activation
CPA 12.5 41 39*** -
# (hours treatment + hours recovery), NQO: 4-Nitroquinoline 1-oxide, CPA: Cyclophosphamide, Blank: not tested
†: Mitotic inhibition (%) = [1 - (mean MIT/mean MIC)] x 100% (where T = treatment and C = negative control), ††: This dose showed mitotic accumulation, whereby the treated mitotic index was 312% of the solvent control, ***: p<0.001
2.6.7. 毒性試験概要表