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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2015, Vol. 9 Issue (1) : 82-89     DOI: 10.1007/s11684-015-0376-0
RESEARCH ARTICLE |
Reproductive toxicity study with a novel deoxyguanosine analogue (Metacavir) in pregnant SD rats
Qihui Luo1,2,Zhengli Chen1,2,*(),Anchun Cheng1,2,Mingshu Wang1,2,Jing Fang1,Xi Peng1,Li Tang1
1. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Ya’an 625014, China
2. Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China
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Abstract  

Our preliminary studies demonstrated that Metacavir has potential to become a new anti-HBV agent. The main targets of the toxic effects of Metacavir, in rhesus monkeys, were gastrointestinal tracts, liver, blood, and kidneys, which were not related to mitochondrial effects. In this study, the maternal toxicity, embryo-fetal developmental toxicity and teratogenicity were studied in pregnant Sprague-Dawley rats after intragastric administration of Metacavir (200, 100, 50, 0 mg/kg body weight) during the first 6–15 days of pregnancy. Slower weight gain was observed in 5 out of 21 rats subjected to a 200 mg/kg dose, as well as 2 out of 20 subjected to a 100 mg/kg dose. Compared with the solvent control group, the calibration weight gain in the 200 mg/kg and 100 mg/kg dosage groups respectively, during first 6–20 pregnant days were significantly different (P<0.01, P<0.05). Significant dose related adverse effects to other reproductive parameters were not seen in F0 and F1, but the number of stillbirths in high dose group showed notably difference compared with the control group (P<0.05), while the litter incidence showed no difference. No Metacavir-associated pathological changes were observed. The present research indicated that at a dose of 200 mg/(kg·d) (i.e., 40 times the effective dose in rats), Metacavir shows some maternal toxicity to SD rats. The embryotoxicity in the 200 mg/kg group encompass decreased fetal body weight, and higher fetal mortality rates, compared with the control group. However, the litter incidence showed no statistical difference. All the treated rats displayed normal bone development, no teratogenicity and without adverse effects on fetal development, thus indicating that below a dose of 200 mg/(kg·d) there is no teratogenic side effects.

Keywords deoxyguanosine analogue      Metacavir      pregnancy      maternal toxicity      embryo toxicity      teratogenicity     
Corresponding Authors: Zhengli Chen   
Just Accepted Date: 06 November 2014   Online First Date: 17 December 2014    Issue Date: 02 March 2015
URL:  
http://academic.hep.com.cn/fmd/EN/10.1007/s11684-015-0376-0     OR     http://academic.hep.com.cn/fmd/EN/Y2015/V9/I1/82
Days of pregnancy 0 mg/(kg·d)(n = 20) 50 mg/(kg·d)(n = 20) 100 mg/(kg·d)(n = 20) 200 mg/(kg·d)(n = 21)
0 227.65±16.28 225.00±18.02 225.74±15.69 225.10±16.90
3 242.00±20.41 240.15±21.68 237.79±15.65 238.90±20.30
6 256.20±24.37 252.30±26.12 246.68±18.24 249.90±23.12
10 273.85±27.85 267.15±29.57 256.68±18.78 260.00±24.16
13 290.15±29.12 283.10±31.14 272.74±24.09* 271.70±25.41
16 309.35±34.29 302.05±33.37 292.53±23.02 289.85±27.74
20 356.50±43.60 352.65±39.88 344.74±28.03 336.70±39.07
6-20 weight gain 100.30±25.98 100.35±17.04 98.05±20.01 86.80±24.35
Calibration weight gaina 40.22±14.88 32.94±9.12 30.56±12.31* 23.58±15.29**
Tab.1  The mean weight and change of weight of F0 female rats (g)
Organ 0 mg/(kg·d)(n = 20) 50 mg/(kg·d)(n = 20) 100 mg/(kg·d)(n = 20) 200 mg/(kg·d)(n = 21)
Terminal weight 270.11±40.2 268.21±44.4 262.86±45.5 251.15±50.5
Liver 3.51±0.25 3.69±0.32 3.36±0.31 3.62±0.42
Spleen 0.34±0.06 0.37±0.09 0.36±0.06 0.31±0.06
Ovaries 0.0231±0.0071 0.0248±0.0052 0.0207±0.0073 0.0255±0.0075
Uterus 0.17±0.04 0.22±0.09 0.20±0.05 0.18±0.08
Pituitary 0.0078±0.0002 0.0069±0.0001 0.0070±0.002 0.0068±0.0001
Tab.2  Mean organ weights relative to terminal body weight (g/100 g body weight)
0 mg/(kg·d) 50 mg/(kg·d) 100 mg/(kg·d) 200 mg/(kg·d)
Pregnant rats number n 20 20 20 21
Abortion number n 0 0 1 1
Premature birth number n 0 0 0 0
F0 female rats death number n 0 0 0 0
Pregnant rats number n 20 20 19 20
Female rats with live births n 20 20 19 20
% 100 100 100 100
Female rats with all resorbed fetal n 0 0 0 0
% 0 0 0 0
Female rats with resorbed fetal n 2 2 1 2
% 10 10 5 9.5
Female rats with stillbirths n 0 0 0 1
% 0 0 0 5
Corpus luteum number Total 272 270 257 263
Corpus luteum of each rat Mean±SD 6.80±2.59 6.75±1.92 6.76±1.73 6.58±2.67
Nidation number Total count 204 220 217 233
Nidation of each rat Mean±SD 5.10±2.54 5.50±1.91 5.71±2.03 5.83±2.44
Stillbirths number n 0 0 0 6*
Death rate of fetal rats % 0 0 0 2.71*
Total resorbed fetal number n 2 2 1 2
Resorbed fetal of each rat Mean±SD 0.1±0.31 0.1±0.31 0.05±0.23 0.11±0.32
Early resorbed fetal number n 1 0 0 2
Early resorbed fetal of each rat Mean±SD 0.05±0.2236 0±0.1 0±0 0.1053±0.3153
Late resorbed fetal number n 1 2 1 0
Late resorbed fetal of each rat Mean±SD 0.05±0.2236 0.1±0.3078 0.0526±0.2294 0±0
Live births of each rat Mean±SD 9.80±2.9249 10.85±2.2659 11.37±1.9541 11.15±3.5184
Male litter size n 99 111 112 114
Percentage of male fetuses per litter % 50.51 51.15 51.85 50.67
Female litter size n 97 106 104 111
Percentage of female fetuses per litter % 49.49 48.85 48.15 49.33
Weight of live births Mean±SD 3.87±0.4019 3.82±0.5101 3.85±0.4529 3.59±0.4959
Placenta mean weight Mean±SD 0.53±0.09 0.55±0.08 0.49±0.06 0.46±0.04
Tab.3  The reproductivity of pregnant rat
Drug Dose Fetal rats(n) / litter Brain inspection Kidney inspection
Cerebral ventricle broaden number (n)/ litter Incidence rate (%) Pelviectasis number(n) / litter Incidence rate (%)
Solvent agent 0 92/20 3/3 3.26/15.00 5/3 5.43/15.00
Metacavir 50 mg/kg 105/20 5/3 4.76/15.00 7/5 6.67/25
100 mg/kg 102/19 3/3 2.94/15.80 5/4 2.94/16.67
200 mg/kg 108/20 2/2 1.85/10.00 7/4 5.55/20
Tab.4  The visceral abnormalities of fetal rats
Drug Dose Inspected pregnant rat number (n) Variable pregnant rat number(n) Incidence rate (%) Inspected fetal rat number (n) Variable fetal rat number (n) Incidence rate(%)
Solvent agent 0 20 5 25.00 92 8 8.70
Metacavir 50 mg/kg 20 7 35.00 105 11 10.48
100 mg/kg 19 6 31.57 102 7 6.86
200 mg/kg 20 6 30.00 108 8 7.41
Tab.5  Summary of fetal rat organs abnormalities
0 mg/(kg·d) 50 mg/(kg·d) 100 mg/(kg·d) 200 mg/(kg·d)
Litter number 20 20 19 20
Fetal number 104 112 114 107
Skeletal deformity (fetal number/ litter) 0/0 0/0 0/0 0/0
Wave costa (fetal number/litter) 0/0 0/0 0/0 0/0
Skeletal variation (fetal number/litter) 48/13 52/15 51/11 50/14
Whole cranium dysostosis (fetal number/litter) 0/0 0/0 0/0 0/0
Nasal bone dysostosis (fetal number/litter) 1/1 2/2 2/1 3/2
Parietal bone dysostosis (fetal number/litter) 4/2 7/3 5/3 5/2
Inca bone dysostosis (fetal number/litter) 0/0 1/1 0/0 1/1
Inferior maxilla dysostosis (fetal number/litter) 0/0 0/0 0/0 0/0
Lingual bone dysostosis (fetal number/litter) 0/0 0/0 0/0 1/1
Upper-hind skull dysostosis (fetal number/litter) 28/12 30/14 26/10 29/13
Cartilago ensiformis dysostosis (fetal number/litter) 16/7 15/7 14/8 18/8
Vertebra thoracalis dysostosis (fetal number/litter) 15/6 17/8 18/7 18/6
Centrum deform (fetal number/litter) 9/5 11/7 11/6 12/7
Centrum not ossified or dysostosis (fetal number/litter) 3/2 5/4 2/2 5/3
Hemivertebrae (fetal number/litter) 0/0 0/0 0/0 0/0
Arcus vertebrae dysostosis (fetal number/litter) 2/2 4/3 1/1 5/3
Supernumerary ribs (fetal number/litter) 0/0 0/0 0/0 0/0
14 costal bone (fetal number/litter) 6/4 5/4 8/5 5/4
Manubrium sterni dysostosis (fetal number/litter) 1/1 2/1 0/0 3/2
First and third sternebrae not ossification or dysostosis (fetal number/litter) 0/0 0/0 0/0 2/2
Last sternebrae dysostosis (fetal number/litter) 21/11 20/12 18/10 23/12
Ossa pubis dysostosis (fetal number/litter) 0/0 0/0 0/0 1/1
Clavicula dysostosis (fetal number/litter) 3/3 1/1 4/2 3/2
Omoplata dysostosis (fetal number/litter) 1/1 0/0 0/0 1/1
Tab.6  Embryo-fetal toxicity and teratogenicity after intragastric administration Metacavir in SD rat (summary of fetal skeletal variation data)
0 mg/(kg·d) 50 mg/(kg·d) 100 mg/(kg·d) 200 mg/(kg·d)
Cervical vertebra 7.00±0.00 7.00±0.00 7.00±0.00 7.00±0.00
Vertebra thoracalis 13.00±0.00 13.00±0.00 13.00±0.00 13.00±0.00
Lumbar vertebrae 6.00±0.00 6.00±0.00 6.00±0.00 6.00±0.00
Sacral vertebrae 3.00±0.00 3.00±0.00 3.00±0.00 3.00±0.00
Caudal vertebrae 4.56±0.57 4.69±0.47 4.62±0.52 4.59±0.58
Costal bone 13.06±0.23 13.05±0.23 13.08±0.27 13.09±0.28
Manubrium sterni 1.00±0.00 1.00±0.00 1.00±0.00 1.00±0.00
Sternebrae 3.85±0.40 3.78±0.41 3.82±0.39 3.80±0.44
Cartilago ensiformis 1.00±0.00 1.00±0.00 1.00±0.00 1.00±0.00
Metacarpal bone 3.77±0.49 3.78±0.53 3.73±0.63 3.68±0.61
Digiti manus 4.63±0.59 4.63±0.57 4.69±0.78 4.64±0.55
Tarsal bones 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00
Metatarsal bone 4.13±0.33 4.15±0.36 4.16±0.37 4.12±0.33
Digiti pedis 4.19±0.50 4.13±0.34 4.17±0.55 4.14±0.35
Tab.7  Number of fetal ossification point after intragastric administration Metacavir in SD rat
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