Hamilton H. Anderson

Antituberculous Activity and Toxicity of Lupulon for the Mouse.

Summary Lupulon, a fat-soluble antibiotic derived from hops, has relatively low in vitro activity (1:40,000) as compared with other antituberculous agents. Despite this observation, lupulon (given orally or intramuscularly) was active against experimental mouse infections of M. tuberculosis. Following intramuscular administration, significantly lower numbers of acid-fast organisms occurred in lesions of treated animals. The approximate numbers relative to the control were: In the liver, 34 to 1; heart, 8 to 1; spleen, 4 to 1; and lungs, 4 to 1; but not in the kidneys. In orally treated animals the ratios were: Liver, 10 to 1; heart, 3 to 1; spleen, 10 to 1; lungs, 5 to 1, and kidneys, 2 to 1. The over-all difference was a reduction of approximately 4 to 1 by either route of administration. Lupulon given intramuscularly within its effective range produced some foci of degeneration in renal tubules. Such pathologic changes were not observed in infected animals given this antibiotic orally in effective doses. The single LD50 on intramuscular administration was 600 mg per kilo in mice; on oral application, 1,500 mg per kilo.

Excretion of Carbon-14 by Man after Administration of Morphine-N-Methyl-C14∗:

Summary 1. Therapeutic doses of morphine-N-methyl-C14 have been administered to 5 human subjects. The major portion of the radioactivity is excreted in 24 hours. Urinary excretion accounted for most of the recovered activity, and there was some correlation between urine volumes and carbon-14 excretion when these volumes were abnormally high or low. 2. Fecal excretion accounted for 7 to 10% of the dose and the possibility that this material originates in the bile as “bound” morphine with subsequent hydrolysis in the intestine is discussed. Intestinal excretion is not as important in man as in rats. 3. Pulmonary excretion of C14O2 ranged from 31/2 to 6% of the injected dose in 24 hours. There was no sex difference in excretion by this route, as has been observed in the rat. 4. The carbon-14 excretion pattern for the one addict was similar to the nontolerant subjects, except in the case of the feces. 5. It has been shown that it is feasible to do tracer studies in normal subjects using less than a microcurie of carbon-14.

Lactic and succinic acid formation by Endamoeba histolytica in vitro.

Abstract Conditions have been developed for obtaining resting trophozoite suspensions of amebae relatively free of associating organisms, which permit study of the fermentative activity of E. histolytica. Thus far, the formation of lactic and succinic acids as fermentation products of E. histolytica has been detected. The Qlactate was observed to be on the order of magnitude of 1 μM 10 × 10 6 cells/hour . The amount of lactic acid formed, of which endogenous activity contributes a significant part, is influenced considerably by the addition of glucose, and also depends upon the culture from which the amebae were derived.

Effects of the Halogenation of Oxyquinoline on Biological Activity.

Chiniofon, N. N. R., introduced commercially as “yatren”, and first tried clinically in amebiasis by Mühlens and Menk 1 in 1921, has had some popularity as an amebicidal agent. Chemically chiniofon is sodium-iodoxy-quinoline-sulphonate, and is related to chinosol N. N. R. (oxyquinoline sulphate) and vioform N. N. R. (iodo-chloroxyquinoline). Since chiniofon has been claimed to have amebicidal activity, we thought it might be of interest to study it from this standpoint in comparison with as many related compounds as we could secure. Such an effort we thought might yield significant information regarding the relation of chemical constitution to pharmacological action in this series of oxyquinoline derivatives, particularly with reference to the effects on biological activity of introducing various halogens into the oxyquinoline molecule. We secured the following compounds for this investigation: 1. Oxyquinoline, from Dr. J. V. Barrow. 2. Oxyquinoline sulphate (Chinosol, N. N. R.). 3. Chloroxyquinoline, from the Ciba Co., Inc. 4. Sodium-iodoxyquinoline sulphonate (Chiniofon, N. N. R.). 5. Iodochloroxyquinoline (Vioform—Ciba, N. N. R.). 6. Diethylamino-dimethylene-hydroxy-iodochlorquinoline HCl, from the Ciba Co., Inc. In this series, compounds 3 and 4 are monohalogenated derivatives of oxyquinoline, while in compounds 5 and 6, two different halogen atoms have been placed in the oxyquinoline molecules. These substances have been studied with respect to toxicity on oral administration to guinea-pigs, rabbits, and cats; balanticidal action in naturally infested guinea pigs; amebicidal action in vitro, and therapeutic effect in monkeys naturally infested with intestinal parasites. The techniques followed have been previously described. 2 A general summary of part of this study appears in Table I.

Evaluation of some chemotherapeutics against Endamoeba histolytica in cultures with Trypanosoma cruzi

Abstract Ten antibiotics and amebacidal agents have been re-studied for their “amebacidal” end points. These determinations were obtained by use of E. histolytica plus T. cruzi and were compared with the end points previously obtained by others utilizing “open” and “sealed” culture technics of E. histolytica plus organism ‘ t. ’. The end point ranges corresponded closely with those obtained with the “sealed culture” technic. The cultures of E. histolytica growing in association with T. cruzi alone provide an opportunity for study of direct amebacidal action in vitro , without the complication that associated bacterial flora has introduced in previous methods.

Acute Toxicity of Trypan Blue, Gentian Violet and Brilliant Green.∗

Difficulties encountered in administering sufficient amounts of the usual chaulmoogra oil derivatives uniformly to effect a cure in leprous subjects have led to the attempted application of other forms of therapy. Thus, recent clinical developments 1 , 2 again have aroused interest in the efficacy of certain non-chaulmoogry (Link Missing 1) drugs, synthetic dyes in particular, against leprosy. In the course of examining the possible antileprotic activity of certain such compounds by a standardized technique, 3 determination of their toxicity in the test animals was a necessary preliminary. As trypan blue, gentian violet, and brilliant green have been used in an empirical manner in the clinic before a critical pharmacological evaluation of their worth has been made, it is advisable to present the results of this toxicity study on experimental animals, before toxic doses may unavoidably be given to human leprous subjects. The intravenous dosage of these dyes, as reputedly used by Ryrie 1 and Ryles, 2 is within the possible lethal range as estimated from experimental animals. While these workers do not give the sources of the dyes used by them, nor their standards of purity, so that it cannot be judged whether or not their dyes are comparable in purity to those used by us, it is significant that Ryrie 1 admits the occurrence of dangerously severe acute toxic effects in his patients. Previously reported toxicity data on these 3 dyes are so inadequate and variant that a recent encyclopedic work on chemotherapy 4 omits all reference to their toxicity. Discrepancies in some cases may be explained by variations in purity and by the use of old solutions. Photodynamic action is not a significant factor in the toxicity of these dyes as far as we could determine. We will later make a more extended report surveying the therapeutic effectiveness as well as the chronic and acute toxicity of these agents. Such a study is indicated in view of enthusiastic endorsement of dye therapy in leprosy on grounds that it is as good as treatment with hydnocarpus oil derivatives, safe, cheap and not unduly painful.

Comparative Biological Activity of Seven New Water-soluble Chaulmoogric Acid Derivatives.

Rogers 1 introduced the first water-soluble chaulmoogra derivative intended for intravenous injection in leprosy, which was recognized to be a systemic disease most rationally treated by administering drugs for systemic effect. Due to the hemolytic, local irritative, sclerosing and other undesirable properties of sodium gynocardate, its intravenous use in humans was generally abandoned even though promising results had been obtained in a considerable number of lepers. No other soluble chaulmoograte has been developed sufficiently free from the disadvantages of “Alepor (sodium hydno-carpate) or sodium gynocardate to warrant its continued intravenous use. This report deals with several compounds which appear from various theoretical considerations to be preferable to the chaulmoogra soaps. Seven new water-soluble derivatives of chaulmoogric acid† have been examined pharmacologically and 3 of these have been tested further by a standardized method for antileprotic efficacy as measured by their influence on the course of an experimental leprous infection in rats. The strain of rats, infection, the time and route of administering the drugs, the diet, size of treated and untreated groups, and all other controllable experimental conditions were identical with those used in work previously described. 2 Further experimental treatment is being continued for a more extended period with the more promising agents. In Table I may be found the tolerated and minimum lethal doses on subcutaneous and intravenous injections in rats, the in vitro bactericidal concentration and the effects in experimental rat leprosy of K-iodo-dihydrochaulmoograte (No. 661-K), Na-chaulmoogryl-glycinate (No. 1141) and Na-dichaulmoogryl-β-glycerophosphate (Drug “A”) 3 in comparison with those of “Alepol” (No. 104). The average variations in body weight, size of leprous lesion and incidence of sloughing lesions in each group of infected rats during a 6-months treatment period are included. The toxicity and in vitro bactericidal activity of Na-chaulmoogryl-o-aminobenzoate (No. 1601), 4 diethyl-ethanolammonium chaulmoograte (No. 2001), choline chaulmoograte (No. 2211) and Na-chaulmoogryl glutamate (Drug “B”) are also shown. Summary. On the basis of in vitro bactericidal and hemolytic action, intravenous toxicity, tolerance, and effectiveness in experimental rat leprosy, Na-dichaulmoogryl-β-glycerophosphate (“chaul-phosphate” for convenience), appears superior for possible use in the intravenous treatment of leprosy than other representative water-soluble chaulmoogra derivatives, such as Na-hydnocarpate (“Ale-pol”), K-iodo-dihy dr ochaulmoograte, Na-chaulmoogryl-glycinate, Na-chaulmoogryl-o-aminobenzoate, diethyl-ethanolamrnonium chaul-moograte, and choline chaulmoograte.

Dependence of Entamoeba histolytica upon associated streptobacillus for metabolism of glucose

Abstract E. histolytica (F-22, streptobacillus associate) contains relatively large amounts of glucosamine. The amoeba cannot synthesize glucosamine, but is dependent upon the bacterial associate for its supply of this aminosugar. This amoeba does not utilize glucose as such, but only the products of glucose metabolism of the mono-associate. A definite metabolic link is believed to occur between this amoeba and its associated organism.

Evidence of direct amebicidal action of agents tested in vitro against Endamoeba histolytica.

Until very recently in vitro studies of amebacides and antibiotics against Endamoeba histolytica were performed with the ameba growing in association with one or more bacteria. This association creates the problem of indirect action of these drugs against the bacterial flora, which in turn may markedly reduce the amebic population. It has been postulated by Anderson and Anderson (1950) that perhaps some of the antibiotics act indirectly by inhibiting the bacterial flora of the intestinal tract. Balamuth and Brent (1949) reported direct amebacidal action of prodigiosin, based on the fact that the amebacidal agent did not affect the growth of the bacterial associate.

Oral Toxicity of Certain Alkyl Resorcinols in Guinea Pigs and Rabbits.

Two alkyl resorcinols have recently been discussed in relation to the treatment of certain parasitic infestations: Lamson et al 1 suggesting hexylresorcinol in ascariasis and uncinariasis, and Faust 2 and Ratcliffe 3 proposing heptylresorcinol (“di-hydranol”) particularly in amebiasis. We believe that clinical trial of new drugs in man may proceed with greater satisfaction than otherwise, if it is made after a critical study of the toxicity of such agents in various genera of mammals for the purpose of reaching an approximate quantitative estimate of the toxic range of the materials, and of determining what pathological effects for which it may be expedient to watch when used clinically and against which to guard. The data presented by Leonard and his associates, 4 while indicating that the toxicity of these drugs is “clinically negligible,” do not seem to have been derived from a critical quantitative study. There is, however, a more significant reason from theoretical grounds for investigating quantitatively the toxicity of the alkyl resorcinols; in many series of related compounds containing alkyl radicals, it has been observed that toxicity in the series increases with increase in the number of carbon atoms in the straight carbon chain, and it is desirable to ascertain to what extent these observations may be expanded into a “law” relating chemical constitution to pharmacological action.