A series of novel aromatic derivatives based on the structure of methylglyoxal bis(guanylhydrazone) (MGBG) was examined for in vitro antitrypanosomal activities and cytotoxicities for human cells. One-third of the compounds tested showed trypanocidal activity at concentrations below 0.5 microM after an incubation period of 72 h. Structure-activity analysis revealed that bicyclic compounds with homocyclic rings and unmodified termini were the most active compounds. Results obtained in three laboratories employing different methods and trypanosome populations consistently ranked compound CGP 40215A highest. This compound had a 50% inhibitory concentration of 0.0045 microM for Trypanosoma brucei rhodesiense, was also active against other trypanosome species, including a multidrug-resistant Trypanosoma brucei brucei, and was significantly less toxic than other compounds tested for a human adenocarcinoma cell line, with a 50% inhibitory concentration of 1.14 mM. The effect of CGP 40215A was time and dose dependent, and low concentrations of the compound required exposure times of > 2 days to exert trypanocidal activity. Compounds were inactive against Leishmania donovani and Trypanosoma cruzi amastigotes in murine macrophages in vitro.

We have measured anaerobic and aerobic energy metabolism in leukocytes from human and animal blood, using several methods. We have taken special care to correct our results for an unavoidable contamination with thrombocytes, which has been neglected in all previous work. White cells were isolated by a method combining sedimentation through Dextran with low speed centrifugations. A pellet of thrombocytes was prepared in parallel from the same blood. Mitochondria were isolated by differential centrifugation in 0.25 M sucrose. Firstly, we have specially studied two cytosolic glycolytic enzymes, phosphoglycerate kinase and pyruvate kinase, both of which catalyze a reaction producing ATP. These two enzymes are very active in leukocytes. Other nucleotides, such as CDP, GDP and UDP can also be phosphorylated. Secondly, we have measured the activity of several mitochondrial enzymes, the most active being α-glycerophosphate oxidase. Using an enzymatic test, we have measured the amount of ATP produced in oxidative phosphorylation; in these experiments it was necessary to consider the activity of adenylate kinase as well as correcting for thrombocyte contamination. Several metabolites in direct or indirect relation with the Krebs cycle were used as oxidizable substrates: glutamate, succinate and α-glycerophosphate gave the best results. Except for amytal, inhibitors of oxidative phosphorylation also inhibit ATP production in leukocyte mitochondria. In conclusion, it can be said that oxidative phosphorylation exists in leukocytes, but that the phosphorylation steps in glycolysis produce 1,000 to 10,000 times more ATP than aerobic energy metabolism.

Two isomeric amidino-2-acetylpyridine amidinohydrazones, 11 and 12, and 4-amidinoindanone amidinohydrazone, 17, have been synthesized and tested for inhibition of S-adenosylmethionine decarboxylase (SAMDC) and diamine oxidase and for antiproliferative activity against T24 human bladder carcinoma cells. Compound 11 inhibited SAMDC with an IC50 of 10 nM and was 140- and > 500-fold more potent than methylglyoxal bis(guanylhydrazone) (MGBG) and 12, respectively. The difference in potency between 11 and 12 was interpreted with the help of molecular modeling and appeared to be associated with two different low-energy conformations of the compounds. Compound 17 which represents a conformationally constrained analogue of 11, was superior to the latter and MGBG with respect to selective inhibition of SAMDC and antiproliferative activity, and is of interest as a potential anticancer agent and a drug for the treatment of protozoal and Pneumocystis carinii infections.

Analogues of 3-aminooxy-1-propanamine proved to be highly potent and selective inhibitors of ornithine decarboxylase (ODC). The compounds competed with ornithine for the substrate binding site of ODC, but resulted in progressive and apparently irreversible inactivation of the enzyme. Diamine oxidase was inhibited by these compounds to a lesser extent than ODC; the compounds were not metabolized by this enzyme. Several derivatives were growth-inhibitory for human T24 cells and for other mammalian cells, the most active compound being 3-aminooxy-2-fluoro-1-propanamine (AFPA). Growth-arrested cells were largely depleted of putrescine and spermidine. Cellular growth arrest could be antagonized by supplementation with spermidine. Selection for resistance against AFPA led to cells with amplified ODC genes and overexpression of the message. Some of the derivatives were tumoristatic at well-tolerated doses in mice bearing solid T24 tumours. The antiproliferative activity of these compounds appears to be mediated by polyamine depletion.

Zymosan stimulated oxygen metabolism was investigated in polymorphonuclear leucocytes (PMN) from 6 diabetic patients. Oxygen uptake and superoxide production were continuously measured in the presence of autologous or control serum and non-opsonized zymosan, or in the absence of serum and preopsonized zymosan. The only significant impairment in the diabetic cells studied was a lower oxygen uptake in the presence of autologous serum. This defect was normalized by addition of control serum or by omitting the serum and stimulating with opsonized zymosan. In the absence of serum, the oxygen consumption was markedly diminished in only one subject, whereas two subjects showed a decrease of superoxide production in the presence of control serum. An inverse correlation between fasting glucose concentration and oxygen uptake could be demonstrated. However, exposure of normal PMN to hyperglycemic glucose concentration in vitro did not significantly alter their oxygen metabolism, suggesting that glucose alone could not be the only factor responsible for the impaired oxygen consumption in diabetic cells.