Metabolism and pharmacokinetics of [14C]-deoxyfructosylserotonin creatinine sulfate administered orally and intravenously to rats and mice.

Deoxyfructosylserotonin (DFS) has been shown in in vitro tests to inhibit L-DOPA-oxidase and also to suppress the multiplication of Mycobacterium leprae. The possible therapeutic use of DFS makes necessary the study of its metabolic fate in animal models. Labelled [14C]-DFS was synthesized by condensation of serotonin and [14C]-glucose and administered per os or intravenously to rats and mice. After oral administration, some of the radioactivity transited through the intestinal tract to be excreted in feces (20-60% of the dose) and some was destroyed in the pH conditions of the intestine and further metabolized by the flora, producing 14CO2 in the expired air (10-40% of the dose). Radioactivity excreted in the urine amounted to 8-15% after 24 h. After intravenous administration, 60-90% of the dose had already been excreted in the urine after 8 h. Feces and CO2 accounted for 5-10% each. In the urine, for both routes of administration, beside DFS, half of the radioactivity corresponded to the glucuronide conjugate, while in the feces all the radioactivity found was unchanged DFS. Whole animal body autoradiography showed the presence of radioactivity in all the organs (1-2% of the dose) mainly resulting from the incorporation of labelled carbon from glucose and CO2. These results, obtained in healthy rats, demonstrate poor intestinal absorption of DFS (10% of the dose) and when it is absorbed, rapid urinary excretion. For its possible therapeutic use as an anti-leprosy drug in humans, derivatives with higher bioavailability must be attained.

Effects of a derivative of serotonin (deoxyfructoserotonin) and other antileprosy drugs on attachment and uptake of Mycobacterium leprae by Schwann cells in vitro.

The association (attachment and/or uptake) of Mycobacterium leprae with cultured Schwann cells was studied at 8 and 72 h in the presence of a new antileprosy compound, deoxyfructoserotonin (DFS), as well as conventional antileprosy drugs such as rifampin (RFP) and 4,4'-diaminodiphenyl sulfone (DDS). DFS significantly inhibited bacterial association with Schwann cells at 8 h. RFP also affected the association of M. leprae but not to the same extent as DFS. A similar inhibition at 8 h was noted when M. leprae but not Schwann cells were pretreated with DFS or RFP for 5 days before infection of cultures, implying that modulation was achieved through some form of drug action on bacteria. DDS had no effect on M. leprae association; however, the combination of DFS and DDS was neither antagonistic nor additive. At 72 h postinfection, when attached but noninternalized bacteria were removed with trypsin-EDTA from Schwann cell cultures containing DFS or RFP, a 50% reduction in the number of bacteria in the drug-treated group was obtained as compared with the numbers in drug-free cultures. This indicated a slow entry of M. leprae into Schwann cells in the presence of these drugs. Collectively, these observations point to differing requirements for late and early association of M. leprae with Schwann cells, besides suggesting a role for DFS and RFP in the prevention and minimization of M. leprae-induced nerve damage in vivo.

[Desoxyfructo-serotonin: its therapeutic effect in the treatment of leprosy]. / La desoxyfructo-serotonine: son effet thérapeutique dans le traitement de la lèpre.

Desoxyfructo-serotonin (DFS) has shown good results in clinical trials of LL patients. After clinical trials in Bamako (Mali) reported in three articles, clinical trials began in India, at Bombay. Acute toxicity tests done in Paris and chronic toxicity tests done in India had shown absence of side effects. This was also confirmed after pre-clinical pharmacology. In vitro tests show that DFS enhances cellular immune response. Receptors for anti-erythrocyte antibody on LL macrophages are demonstrated by erythrocyte rosetting. Infection with M. leprae markedly reduces rosetting. But in the presence of DFS this reduction in rosetting is not observed. Patient's peripheral blood lymphocytes, sensitised with leprosy antigen, show a low level of rosetting with patients' macrophages. DFS greatly enhances the lymphocyte-macrophage interaction. DFS has an important anti-stress activity. Gastric ulcer induced in rats by restraint were reduced by 40% (Mester et al.) and 50% (Das Neves). DFS increased the uptake of serotonin by LL patients platelets. HPLC studies were done to see the level of DFS in the plasma, in the serum and in the urine of LL patients and controls. We are synthetising new lyposoluble derivatives in order to make easier the penetration of DFS and a long time effect.

Effect of deoxyfructoserotonin (DFS) on lepromatous leprosy.

To examine the anti-leprosy effect of deoxyfructoserotonin the drug was given in a dose of 10 mg/kg for 6 months to 6 patients with active lepromatous leprosy, in accordance with the WHO-THELEP protocol. Clinical and histological assessment and mouse foot-pad studies suggest that the drug has some anti-leprosy effect.

[Treatment of leprosy by human metabolites]. / Traitement de la lèpre par des métabolites humaines.

We are interested for other human metabolites than desoxyfructo-serotonin (DFS), showing antileprosy activity. This is the case of desoxyfructo-5-hydroxytryptophan and of some liposoluble derivatives of DFS. The time of resorption and penetration into M. leprae infected tissue, is very different for these metabolites. For this reason the simultaneous application of these compounds may represent some advantage in the treatment of multibacillar form of leprosy. The use of DFS together with the antileprosy diet "NAL" have the supplementary advantage to stabilize the DFS level in the serum during the treatment.

Mouse foot-pad studies with M. leprae--effect of desoxy fructo serotonin (DFS) and related compounds.

Mouse foot-pad experiments were carried out to study the effects of DFS and related compounds on the multiplication of M. leprae. Of the 25 cases clinically suspected Dapsone resistance, 8 were found resistant and 14 sensitive to Dapsone by mouse foot-pad experiments. Six were resistant to DFS and 16 were sensitive. Desoxy fructo 5-hydroxy tryptophane as well as Nutrition Antileprosy (NAL) diet were also found effective in suppressing the growth of M. leprae in mouse foot-pad. Of the two liposoluble derivatives of DFS tested (DFS LS-I and DFS LS-II), DFS LS-II was found more effective in suppressing the growth of M. leprae in foot-pads of mice.

Nutritional aspects of leprosy.

Desoxyfructo-serotonin (DFS) being a naturally occurring metabolite, which, like serotonin, has its origin in tryptophane in the food, one can ask about the role of nutrition in leprosy. The unique situation, that a human metabolite shows antileprosy activity, confirmed in vitro and in vivo, makes it possible to develop a "build in" antileprosy therapy. This is now realised through a so-called "Anti-Leprosy Nutriment" (NAL). The effectiveness of this diet was confirmed by the fact, that a daily dose of 0.5 g NAL per mouse has a similar effect, like 20 mg/kg body weight Dapsone per day in the conventionnal mouse foot-pad test. The biosynthesis of DFS in man has been demonstrated by high performance liquid chromatography (HPLC), spectrofluorometry and mass spectrometry. The activity of NAL (rich in tryptophane, unsaturated fatty acids and glucose) is due to an increased biosynthesis of DSF. The latter may be considered as a "physiological protecting agent" against leprosy. This type of food may play a role in the prevention of leprosy in endemic area.

[Action of desoxyfructo-serotonin on Mycobacterium leprae. Results of inoculation of human biopsy specimens in the mouse after a year of treatment]. / Activité de la désoxyfructo-sérotonine sur Mycobacterium leprae. Résultats de l'inoculation de biopsies humaines à la souris après un an de traitement.

M. leprae, extracted from cutaneous biopsies of 4 out of 7 patients treated with DFS during one year at doses of 1000 to 1500 mg/day were injected in mouse foot pads. Bacterial multiplication was observed in each case. The purely bacteriostatic activity of DFS for M. leprae, previously observed in mice, is thus confirmed. The eventual place of DFS in the therapy of leprosy is discussed.

Deoxyfructo-serotonin: a new drug with anti-leprosy activity.

In the mouse foot pas, deoxyfructo-serotonin (DFS) shows a definite inhibitory effect on the multiplication of M. leprae in Dapsone-sensitive as well as Dapsone-resistant cases. In clinical trials, based on 7 cases treated with DFS for an average period of 6 months, the beneficial effects of DFS are observed after a few weeks of treatment in BB cases, and after a few months in LL cases. In addition to clear improvements in the appearance of the skin (regression and healing of nodules, almost complete disappearance of infiltration, etc) one observes in the majority of cases a rapid improvement in the bacteriological and morphological indexes, the latter falling to 0%. In the course of these studies, no single suggestion of intolerance was detected.

Low serotonin uptake by platelets in leprosy and a new approach to prevent it.

The plasma of leprosy patients contains high levels of mucoproteins which are deficient in sialic acid. However, due to the increased mucoprotein level, the total sialic acid content of leprous plasma, calculated on protein, is increased when compared with normal human plasma. The low serotonin uptake observed with isolated platelets is probably due to their low sialic acid content. The inability of normal human plasma to correct the diminished serotonin uptake by isolated leprous platelets is in favor of a definite structural change in leprous platelets, related to their low sialic acid content. In patients with active disease and in those with lepra reactions, leprous plasma itself can correct the diminished uptake of serotonin by the isolated platelets. In patients with subsided lepra reactions, the leprous plasma is much less effective. In severe cases, where serotonin uptake is decreased even in platelet rich plasma, desoxyfructo-serotonin increased the uptake of serotonin.