The importance of the relationship between the energetic centers present in alkylating agents and their ability to produce type D offbalance, led us to study a group of these substances which, at once, have both short molecules and two polar groups in close proximity. The desire to have such an agent with lipoidic property as well led us to study epichlorohydrine which corresponds to propane and has an epoxy group binding C2 and C3, while C1 binds a chlorine. Soluble in neutral solvents, epichlorohydrine becomes soluble in water only after hydrolysis. Its biological activity differs from other chlorohydrines such as chloropropanediol or trichloropropane, both of which can be considered to be closely related to the substance produced by hydrolysis of epichlorohydrine. The acute toxic dose of epichlorohydrine was found to be 6 mgr./30 gr. for mice and 25 mgr./100 gr. for rats by intraperitoneal administration; 22 and 35 mg./100 for mice and rats by subcutaneous injection. In tests for chronic toxicity, it was apparent that doses of 5 and 1.5 mgr. injected daily were well tolerated respectively by rats and mice. With higher doses, the animals became rapidly emaciated before dying. Used orally in drinking water, a solution of 1/3000 was well tolerated by rats and mice even for months. With the use of solutions of 1/2000, only a few animals did not lose weight, while a solution of 1/1000 invariably induced weight loss.

There were no effects observed upon microbes or bacteriophage.

It appears that epichlorohydrine, acting below the morphological levels, induces changes similar to those seen for other alkylating agents. However, it is not upon the desoxyribo nucleic acids present that an important action is seen but in the lipidic system at these lower levels. Epichlorohydrine seems to act also at other levels. The influence upon pain—an increase for alkaline pattern, a decrease for acid—was more noticeable than for sulfur compounds. Delayed effects, however, were more obvious than immediate ones. The influence upon wound healing was similar to that of polyunsaturated fatty acids. Cancerous cells, such as those from mouse ascites, were destroyed in vitro by a 0.5 solution of epichlorohydrine. The effect in vivo upon sarcoma 180 or Ehrlich ascites tumors was most interesting. Administered by subcutaneous or intramuscular injection, epichlorohydrine had no effect on the tumor even in doses as high as 2.5 mgr. daily. However, when administered in drinking water in a 1/1000 solution, it prevented the development of ascites in 19/20 animals. But the toxicity was too high. A 1/2000 solution, used as drinking water, controlled the condition in more than 50% of the animals, while a 1/3000 solution showed favorable resuits in only a few animals. Under the same conditions, there was no apparent effect upon solid tumors in mice, even those induced by subcutaneous injection of ascites tumor cells.

In humans, all effects upon the tumors were interesting and will be discussed below. The influence upon systemic patterns was relatively reduced except for a marked effect upon the elimination of calcium in urine, obtained even with small doses which produced no other changes. Repeated injections of organ cells treated in vitro with epichlorohydrine were able to induce severe degenerative changes in the respective organs. Experiments with tumor cells treated and administered in the same manner are still in progress.