NTP Toxicology and Carcinogenesis Studies of Dimethyl Methylphosphonate (CAS No. 756-79-6) in F344/N Rats and B6C3F1 Mice (Gavage Studies).
Dimethyl methylphosphonate (98% pure) is one of four chemicals nominated by the U.S. Army for toxicology and carcinogenesis studies because it was being considered for use to simulate the physical and spectroscopic (but not the biologic) properties of anticholinesterase (nerve) agents. Dimethyl methylphosphonate is also used as a flame retardant, a preignition additive for gasoline, an antifoam agent, a plasticizer and stabilizer, a textile conditioner and antistatic agent, and an additive for solvents and low-temperature hydraulic fluids. The United States produces 0.2-2 million pounds (91,000-910,000 kg) of per year. Gavage was chosen as the route of administration for all four candidate "simulants" to mimic potential exposure. Experimental Design: Dimethyl methylphosphonate was administered in corn oil by gavage to male and female F344/N rats and B6C3F1 mice in single-administration, 15-day, and 13-week studies to obtain toxicity data, to establish dose levels for the 2-year studies, and to identify target tissues. Additional studies were also performed to determine toxicity to the reproductive system of male F344/N rats and B6C3F1 mice and to study the potential for genetic damage in bacteria, mammalian cells, and Drosophila. Single-Administration Studies: In the single-administration studies, dimethyl methylphosphonate was given to rats and mice at doses up to 6,810 mg/kg body weight. No compound-related deaths were seen in male or female rats or male mice; two high dose female mice died. Rats exhibited inactivity, unsteady gait, and prostration after dosing; mice were inactive after dosing. Fifteen-Day Studies: Rats and mice received doses of 0, 1,250, 2,500, 5,000, 10,000, or 15,000 mg/kg dimethyl methylphosphonate per day. Compound-related deaths occurred in the three highest dose groups of rats and the two highest dose groups of mice. Rats receiving doses of 2,500 mg/kg or higher were inactive and at 5,000 or 10,000 mg/kg had an unsteady gait after dosing; mice exhibited inactivity, shallow breathing, and prostration at doses of 10,000 mg/kg or higher. No lesions were reported in rats. Nonneoplastic lesions of the stomach were seen in some male mice at doses of 1,250 mg/kg and higher and in some female mice at doses of 5,000 mg/kg and higher. Thirteen-Week Studies: Dimethyl methylphosphonate was given at doses up to 8,000 mg/kg per day. Compound-related deaths occurred at 2,000, 4,000, and 8,000 mg/kg in rats and at 4,000 and 8,000 mg/kg in mice. Mean body weights of rats at 1,000 mg/kg and mice at 2,000 mg/kg were similar to those of the vehicle controls; decreased weight gain was seen at higher doses. No compound-related clinical signs were reported. Minimal to mild renal and testicular lesions were seen at all doses in male rats, but the severity of these lesions did not increase with increasing dose of the chemical. No apparent target tissues were identified in female rats or male and female mice. Doses selected for the 2-year studies were based on body weight effects and mortality seen in the 13-week studies; the lesions seen in the kidney of male rats at the end of the 13-week studies were judged not to be life threatening. In the 2-year studies, dimethyl methylphosphonate was administered in corn oil by gavage at doses of 0, 500, or 1,000 mg/kg per day to groups of 50 F344/N rats of each sex and at 0, 1,000, or 2,000 mg/kg per day to groups of 50 B6C3F1 mice of each sex. All animals were dosed 5 days per week for 103 weeks. Body Weight and Survival in the Two-Year Studies: Mean body weights of high dose male rats were 5%-10% lower than those of the vehicle controls between weeks 28 and 76 and were 10%-24% lower between weeks 80 and 104. Mean body weights of high dose female rats were 8%-12% lower than those of the vehicle controls after week 80. Survival of male rats was greater than 50% in all groups until week 80, and after this time, survival decreased in both groups, with the survival at the end of the study being 27/50 in vehicle control, 17/50 in low dose, and 4/50 in high dose groups. Survival of in low dose, and 4/50 in high dose groups. Survival of low dose female rats was comparable to that of the vehicle controls, but the final survival of high dose female rats was decreased (vehicle control, 30/50; low dose, 33/50; high dose, 23/50). No other compound-related clinical signs were observed. Mean body weights of high dose male mice were 7&percnt;-16&percnt; lower than those of the vehicle control males between weeks 36 and 76, and those of high dose female mice were 6&percnt;-12&percnt; lower between weeks 88 and 103. Decreased survival between weeks 23 and 45 in high dose male mice was associated with fighting. Seventeen high dose male and 22 high dose female mice died during week 45; these deaths were associated with the accidental administration of a dose mixture that had a concentration 34&percnt; greater than the targeted amount. Eleven low dose male mice died on the same day during week 77. By the end of the study, 29/50 vehicle control, 12/50 low dose, and 0/50 high dose male mice were alive; 41/50, 30/50, and 2/50 female mice survived to the end of the study. Renal Effects in the Two-Year Studies: Administration of dimethyl methylphosphonate to male rats increased the average severity of nephropathy and caused mineralization (calcification) of the collecting tubules in the renal papilla (12/50; 41/50; 36/49), hyperplasia of the transitional epithelium lining the renal pelvis and overlying the renal papilla (0/50; 23/50; 21/49), and focal hyperplasia of the renal tubular epithelium (0/50; 8/50; 9/49). Administration of dimethyl methylphosphonate to male rats was also associated with the occurrence of rare renal tubular cell adenocarcinomas (0/50; 2/50; 3/49) and papillomas of the transitional epithelium lining of the renal pelvis (0/50; 2/50; 3/49); a transitional cell carcinoma occurred in a low dose male rat. There were no tubular cell or transitional cell neoplasms of the kidney in female rats. Hematopoietic System Effects in the Two-Year Studies: The incidence of mononuclear cell leukemia was increased in high dose male rats (10/50; 11/50; 17/50). Genetic Toxicity: Dimethyl methylphosphonate was not mutagenic when tested in the Salmonella typhimurium/microsome assay by the preincubational protocol with strains TA98, TA100, TA1535, or TA1537 with or without metabolic activation. The chemical did induce forward mutations in the mouse lymphoma L5178Y/TK&plusmn; assay system in the absence of metabolic activation. Treatment of cultured Chinese hamster ovary cells with dimethyl methylphosphonate did not induce chromosomal aberrations; however, sister chromatid exchanges were induced after exposure to this chemical in both the presence and absence of metabolic activation. When fed to Drosophila, dimethyl methylphosphonate induced a significant increase in the frequency of sex-linked recessive lethal mutations but did not induce reciprocal translocations. Dimethyl methylphosphonate caused a dominant lethal effect in male rats and mice. Studies of Reproductive Effects: Dimethyl methylphosphonate caused a dose-related increase in the number of fetal resorptions in undosed female rats and mice mated with males that received the chemical by gavage in water 5 days per week for 13 weeks at doses of 0-2,000 mg/kg per day. After the 13-week dosing period, histopathologic changes were seen in the kidney and testis of male rats but not in male mice; dosed male rats sired fewer litters and fewer pups per litter. Dose-related decreases in sperm count and sperm motility occurred in male rats but not in male mice. Toxic effects to the reproductive system of male rats and mice were reversible after a 13-to 14-week recovery period. Data Audit: An audit of the experimental data was conducted for the 2-year studies on dimethyl methylphosphonate. No data discrepancies were found that influenced the final interpretations. Conclusions: Under the conditions of these 2-year gavage studies, there was some evidence of carcinogenic activity of dimethyl methylphosphonate for male F344/N rats as shown by increased incidences of tubular cell hyperplasia, tubular cell adenocarcinomas, hyperplasia of the transitional cell epithelium, and transitional cell papillomas of the kidney. There was an increased incidence of mononuclear cell leukemia in male rats at 1,000 mg/kg. Renal toxicity and decreased survival occurred in dosed male rats. There was no evidence of carcinogenic activity of dimethyl methylphosphonate for female F344/N rats given doses of 500 or 1,000 mg/kg. The study in male B6C3F1 mice was an inadequate study of carcinogenic activity because of decreased survival in both dosed groups. There was no evidence of carcinogenic activity for female B6C3F1 mice receiving dimethyl methylphosphonate at 1,000 mg/kg; decreased survival of female mice at 2,000 mg/kg made this group inadequate for determination of carcinogenic activity. Synonyms: fyrol DMMP; methyl phosphonic acid, dimethyl ester; DMMP; methanephosphonic acid dimethyl ester; dimethyl methanephosphonate