Pregnant ICR-JCL mice, Wistar rats, and Japanese white rabbits were injected intraperitoneally with doses of 0.2 to 80 mg/kg of diltiazem hydrochloride, a coronary vasodilator, on successive days in the period of organogenesis. Further, the drug was given via the same route in a single dose of 3.1 to 80 mg/kg on various days of gestation to pregnant mice and rats from the same colonies used in the above experiments. The animals were sacrificed at term and their fetuses were examined for gross external and skeletal anomalies in cleared specimens stained with alizarin red S. The main results were:
1. Pregnant mothers tolerated well all doses.
2. The fetal mortality rate after the successive injections was significantly high at the dose levels of 12.5 mg/kg and over in mice and rabbits, and at 80 mg/kg in rats as compared with the controls.
3. The successive injections induced an appreciable number of externally malformed fetuses in rats and rabbits at 80 mg/kg and at 12.5 mg/kg, respectively, but not in mice. Malformations of the limbs and tail were the main types of defects observed.
4. A small number of fetuses with vertebral and rib deformities were observed in all of the three species after the successive injections.
5. The single injection experiments revealed that the susceptibility to the embryolethal action suddenly increased from day 9 in mice and day 12 in rats, reached the highest level on days 10 and 11 in mice and on day 12 in rats, and gradually decreased thereafter. On the most susceptible days, the fetal LD50 was estimated to be between 6.3 and 12.5 mg/kg in mice and between 40 and 80 mg/kg in rats.
6. The highest incidence of gross external anomalies induced by the single injection was recorded on day 13 in mice and rats. The lowest teratogenic dose was 25 mg/kg in mice and 80 mg/kg in rats. Limb and tail anomalies predominated.
7. The injection of 25 or 50 mg/kg on day 9 in mice and of 80 mg/kg on day 11 in rats produced vertebral and rib defects.
The relationship between embryolethality and teratogenicity, and possible mechanism of the embryotoxicity are discussed.
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