MX2008004026A - Pharmaceutical compositions containing nitrovinylfuran derivatives for the treatment of leishmaniasis and trypanosomiasis - Google Patents

Abstract

The invention relates to the action of representatives of a novel family of products in relation to different strains ofLeishmaniaandTrypanosoma cruzifor the treatment of cutaneous, mucocutaneous and visceral leishmaniasis as well as trypanosomiasis. The aforementioned products have demonstrated greater effectiveness than medicaments already on the market inin vitrostudies on laboratory animals and humans in relation toLeishmania mexicana amazonensis, L. donovani infantum, L.braziliensis braziliensisandTrypanosoma cruzi. The pharmaceutical effect of formulations of said family is proven for human and veterinary use in the treatment of the above-mentioned diseases using different administration routes.

Description

PHARMACEUTICAL COMPOSITIONS CONTAINING NITROVINILPHURANIC DERIVATIVES FOR THE TREATMENT OF THE LEISHMANIOSIS AND TRIPANOSOMOSIS FIELD OF THE INVENTION The present invention relates to the branch of human and animal health in general with a family of products that have biological activity against the etiological agents that cause leishmaniasis and tripanosomosis.

TECHNICAL BACKGROUND Leishmaniasis is a disease caused by protozoa of the genus Leishmania. This includes around 20 pathogenic species for humans or animals. It is recognized that there are manifestations of this in 88 countries, presenting an annual report of 1.5-2 million new cases, estimating that there are more than 12 million patients in the world and 350 million at risk of contracting the disease each year (CDC. http://www.dpd.cdc.gov/dpdx., Parasitic Disease Information, Fact Sheet 1.April 2004).

Clinically there are three fundamental ways: Cutaneous leishmaniasis: It is the most widespread, representing the 90% of cases and is characterized by ulcerated lesions of the skin that can take from months to several years to heal. It is common for these lesions to be complicated by secondary bacterial or fungal infections. Visceral leishmaniasis: It produces hepato- and splenomegaly, it affects the red bone marrow; It causes anemia, progressive weight loss and death in the majority of untreated cases. Of those who receive treatment late, it dies between 15 and 25%. Mucocutaneous leishmaniosis: A variable time (months-years) after the healing of the primary cutaneous lesion or even coinciding with it, the parasites spread to the oral, nasal and pharyngeal mucous membranes. Destruction of the soft tissues, especially cartilage, occurs, which causes monstrous deformations of the face and compromises the patient's life when breathing is affected by damage to the laryngeal cartilages. Acquired Immunodeficiency Syndrome (AIDS) has aggravated the symptoms of leishmaniasis, since patients require longer treatments and the frequency of relapses is high (Alvar, J; et al., Clin. Microbiol. Rev. 1997 (10): 298 -319.

For more than 50 years pentavalent antimony derivatives have been the drugs of choice. Currently, Pentostam® (Sodium Antimony Gluconate) and Glucantime® (Meglumine Antimoniate) are the pentavalent forms of antimony on the market. These are not effective against all species, L. aethiopica and L. major being particularly unresponsive. Its efficacy against normally susceptible species is not 100%, with an increasing number of strains developing resistance to pentavalent antimony derivatives (Aparicio P., et al., Antiparasitic Therapeutics, Infectious Disease Microbiol.; 21 (10) 579-94). Its use has been limited by adverse effects, frequent cause of treatment interruption. Subclinical pancreatitis (31%), cardiotoxicity (15%) and nephrotoxicity are associated with pentavalent antimony in 5% of cases. Patients with AIDS coinfected with Leishmania are particularly sensitive to pancreatitis, to the point of being considered a potential cause of death in these patients (Alvar, J; et al., Clin.Microbiol, Rev. 1997 (10): 298-319. Amphotericin B and pentamidine are the other two drugs available for the treatment of leishmaniasis, due to their toxicity they are reserved for cases resistant to pentavalent antimony derivatives, and lipid formulations have been developed for amphotericin B (Alphocil® and AmBisone®) with less toxicity and adequate efficacy, but the price is not affordable to the majority of the population affected (Aparicio P., et al., Antiparasitic Therapeutics, Infectious Disease Microbiol Clin 2003; 21 (10) 579-94). American trypanosomosis, whose etiological agent is Trypanosoma cruzi, known as Chagas disease, is a health problem endemic to all of Latin America. This disease produces around 50,000 deaths per year. It is estimated that 20 million people suffer from it, and that another 100 million inhabitants of the continent are at risk of contracting it. The parasite is transmitted to man through the feces of hematophagous insects (Anonymous, http://www.unl.edu.ar/eie.php?ID=1834, September 15, 2005). During the past six decades, more than 100 experimental products with different chemical structures have been tested for the treatment of infections caused by T. cruzi, but only some of them have completed the preclinical phase with relative success. Only two trypanocidal drugs have been accepted, registered and marketed by national health authorities in Latin American countries: nifurtimox (1972-1992), currently eliminated from the market, and benzonidazole available since 1975 (Paulinoa et al., Mini Rev Med Chem 2005: 5 (5): 499-519; Lockman JW, Hamilton AD Curr Med Chem. 2005; 12 (8): 945-59).

DETAILED DESCRIPTION OF THE INVENTION The present invention describes the action of pharmaceutical compositions containing any of the following 6 representatives of the family of β-nitrovinylfurans: 2- (2-nitrovinyl) -furan 2-bromo-5- (2-nitrovinyl) -furan 2 - (2-bromo-2-nitrovinyl) -furan 2-bromo-5- (2-bromo-2-nitrovinyl) -furan, 2- (2-methyl-2-nitrovinyl) -furan 2-bromo-5- ( 2-methyl-2-nitrovinyl) -furan These pharmaceutical compositions include, in addition to the β-nitrovinylfuran derivatives, vehicles, excipients, solvents and adjuvants for pharmaceutical use. The solvents are preferably apolar, such as sunflower oil, petrolatum or Migiyol 810. These compositions include the β-nitrovinylfuran derivatives at concentrations ranging from 0.01 to 10%. The exact composition is a function of the product to be used and the route of administration to be used. The possible routes of administration are topical or systemic, depending on the clinical form of the disease. For visceral and mucocutaneous leishmaniasis and trypanosomosis, it is administered in ways that guarantee a systemic distribution. In contrast, for cutaneous leishmaniasis and other manifestations cutaneous diseases of the diseases referred to above, both are used, that is, the topical and systemic routes. The products were tested against 3 species of Leishmania, representative of the causes of cutaneous, mucocutaneous and visceral forms, as well as against Trypanosoma cruzi, causal agent of Chagas disease.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1: Activity of 2-bromo-5- (2-bromo-2-nitrovinyl) -furan and 2-bromo-5- (2-methyl-2-nitrovinyl) -furan against experimental cutaneous leishmaniosis. Treatment of recent injuries, every 24 h. Figure 2: Activity of 2-bromo-5- (2-bromo-2-nitrovinyl) -furan and 2-bromo-5- (2-methyl-2-nitrovinyl) -furan against experimental cutaneous leishmaniosis. Treatment of recent injuries every 12 h. Figure 3: Activity of 2-bromo-5- (2-bromo-2-nitrovinyl) -furan and 2-bromo-5- (2-methyl-2-nitrovinyl) -furan against chronic experimental cutaneous leishmaniasis during seven first days after the treatment was established. Figure 4: Activity of 2-bromo-5- (2-bromo-2-nitrovinyl) -furan and 2-bromo-5- (2-methyl-2-nitrovinyl) -furan against chronic experimental cutaneous leishmaniasis to completion the treatment.

EXAMPLES OF MODALITY EXAMPLE 1 In vitro activity of ß-nitrovinylfurans against promastigotes of Leishmania The mean inhibitory concentrations (IC50) of the ß-nitrovinylfuran derivatives were determined against promastigotes of L. mexicana amazonensis, L. donovani infantum and L. braziliensis braziliensis. For this, the method described by Bodley et al. Likewise, the minimum parasiticide concentrations (CP100) were determined, these being the minimum concentrations of each product able to cause the immobility of all the promastigotes in the crop. As reference control drugs, amphotericin B and Glucantime® were also evaluated. All the products showed an inhibitory effect of the growth of promastigotes at considerably low doses. Likewise, the lethal effect on the parasites occurred at low concentrations and from the first two to three hours after the products were added to the culture. The results also showed that amphotericin B was more active than ß-nitrovinylfurans, while Glucantime® showed less activity than these.

TABLE 1 In vitro effect of ß-nitrovinylfurans against promastigotes of L. m. amazonensis, L. b. braziliensis and L. d. infantum Legend A: 2- (2-nitrovinyl) -furan B: 2-bromo-5- (2-nitrovinyl) -furan C: 2- (2-bromo-2-nitrovinyl) -furan D: 2-bromo-5- ( 2-bromo-2-nitrovinyl) -furan E: 2- (2-methyl-2-nitro-vinyl) -furan F: 2-bromo-5- (2-methyl-2-nytrovinyl) -furan EXAMPLE 2 In vitro activity against Trypanosoma cruzi The in vitro action of the products was evaluated against Trypanosoma cruzi trypomastigotes grown in a cell line of rat myoblasts. The minimum parasiticidal concentrations (minimum concentrations that cause the immobilisation of all parasites in the culture) and the average inhibitory concentrations (concentrations that reduce the number of parasites to half of those existing in the untreated control cultures) were determined according to Buckner et al. to the. The test showed that the products have an in vitro parasiticidal effect superior to that of benzonidazole.

TABLE 2 In vitro effect of ß-nitrovinylfuran derivatives against Trypanosoma cruzi Legend A: 2- (2-nitrovinyl) -furan B: 2-bromo-5- (2-nitrovinyl) -furan C: 2- (2-bromo-2-nitrovinyl) -furan D: 2-bromo-5- ( 2-bromo-2-nitrovinyl) -furan E: 2- (2-methyl-2-nitrovinyl) -furan F: 2-bromo-5- (2-methyl-2-nitrovinyl) -furan EXAMPLE 3 Toxicology studies In the same animal model in which the anW-Leishmania activity studies were developed in vivo, studies were conducted comparative toxicology of ß-nitrovinylfuran derivatives, using formulations whose main excipient was sunflower oil and the route of intraperitoneal administration. Female Balb / c mice, 18-20 g, were used, in which the mean lethal doses (LD50) of each one were determined. of the products and of amphotericin B. From the dose-effect curves, the maximum non-fatal doses (DLO) were also estimated. Also I know determined the maximum tolerated doses (DMT, doses that do not provoke mortality or weight loss greater than 10% of the initial weight) by administration for 14 days with intervals of 12 or 24 h. He could check that ß-nitrovinylfuran derivatives have lethal doses means variable and superior to amphotericin B. Maximum doses Tolerances determined allowed to select the doses to be used in the efficacy tests in vivo.

TABLE 3 Mean lethal doses (LD50), maximum non-fatal doses (DLO) and doses tolerated maximums (DMT) of the ß-nitrovinylfuran derivatives and the amphotericin B Legend A 2- (2-Nitrovinyl) -furan B 2 -bromo-5- (2-nitrovinyl) -furan C 2- (2-bromo-2-nitrovinyl) -furan D 2 -bromo-5- (2-bromo- 2-nitrovinyl) -furan E: 2- (2-methyl-2-nitrovinyl) -furan F 2 -bromo-5- (2-methyl-2-nitrovinyl) -furan EXAMPLE 4 Treatment of experimental cutaneous leishmaniasis with 2-bromo-5- (2-bromo-2-nitrovinyl) -furan and 2-bromo-5- (2-methyl-2-nitrovinyl) -furan. Recent injuries treated every 24 hours Balb / c mice were experimentally infected with promastigotes of L. m. amazonensis in the plantar bearings. The products were applied intraperitoneally at a rate of 5 mg / kg for the case of 2-bromo-5- (2-bromo-2-nitrovinyl) -furan and 50 mg / kg for 2-bromo-5- (2 -methyl-2-nitrovinyl) -furan. Amphotericin B was administered at a dose of 1 mg / kg intraperitoneally. The treatments were applied daily for 14 days from six weeks after the animals were inoculated. A group that was administered Mygliol 810 (vehicle used for 2-bromo-5- (2-bromo-2-nitrovinyl) -furan and 2-bromo-5- (2-methyl-2-nitrovinyl) was included. -furano) and a control to which no treatment was applied. The growth curves of the lesions from the beginning of the treatment until 2 weeks after the end of the treatment (graph 1) show that in the treated mice the 2-bromo-5- (2-bromo-2-nitrovinyl) -furan or the 2-bromo-5- (2-methyl-2-nitrovinyl) -furan was produced, in the first week, a minimal growth of the lesions; even a slight decrease is observed in the case of 2-bromo-5- (2-methyl-2-nitrovinyl) -furan. At this time the increases in the lesions of the animals treated with the products were statistically lower than that shown by the rest of the animals. groups As of the second week, the growth of the lesions occurs at a rate similar to that of the controls. The differences established from the point of view of the size of the lesions of the animals treated with 2-bromo-5- (2-bromo-2-nitrovinyl) -furan during the first week remain until two weeks after the end of the treatment. Conversely, the lesions of the animals treated with amphotericin B evolved in a similar way to the controls until the end of the treatment. However, in the first week after finishing the treatment a marked decrease in the growth rate was observed.

EXAMPLE 5 Treatment of experimental cutaneous leishmaniasis with 2-bromo-5- (2-bromo-2-nitrovinyl) -furan and 2-bromo-5- (2-methyl-2-nitrovinyl) -furan. Recent injuries and treatment every 12 h The test was carried out in a manner similar to that described in example 4, except that doses of 2 mg / kg of 2-bromo-5- (2-bromo-2-nitrovinyl) -furan and 100 mg / kg of 2 were applied. -bromo-5- (2-methyl-2-nitrovinyl) -furan. Also, the treatment was applied intraperitoneally, but every 12 h instead of 24 h. During the first week of treatment, lesions of animals treated with 2-bromo-5- (2-bromo-2-nitrovinyl) -furan decreased in size (graph 2), being statistically lower (PO.001) than those of the controls. From this moment, the lesions grew at a rate similar to that of the other groups. During the following week a new decrease in the growth rate of the lesions was observed, which caused that 7 days after finishing the treatment the size of the lesions of the animals treated with 2-bromo-5- (2-bromo-2-nitrovinyl) ) -furano were smaller (PO.05) than those of the control animals. The growth of the lesions of the animals treated with 2-bromo-5- (2-methyl-2-nitrovinyl) -furan or 2-bromo-5- (2-bromo-2-nitrovinyl) -furan was significantly lower than the of the controls not treated. Similarly, in comparison with amphotericin B, both products showed greater activity.

EXAMPLE 6 Treatment of experimental cutaneous leishmaniasis with 2-bromo-5- (2-bromo-2-nitrovinyl) -furan and 2-bromo-5- (2-methyl-2-nitrovinyl) -furan. Chronic injuries treated every 12 hours Eighteen weeks after the mice were infected, they have already developed chronic lesions of leishmaniasis, mostly ulcerated, covered with thick crusts. The mean dorso-ventral diameter of the infected legs was 1.23 ± 0.24 cm (mean ± standard deviation) and the lateral diameter was 1.24 ± 0.19 cm. Under these conditions, treatment with 2-bromo-5- (2-bromo-2-nitrovinyl) -furan was initiated at a dose of 2 mg / kg, twice day; 2-bromo-5- (2-methyl-2-nitrovinyl) -furan at 50 mg / kg at the same frequency; or amphotericin B at a rate of 5 mg / kg every other day. A control group that did not receive treatment was also included. All products were administered intraperitoneally for 14 days. In the first week of treatment, 2-bromo-5- (2-bromo-2-nitrovinyl) -furan caused a decrease in the dorso-ventral diameter of the lesions that differed statistically (P <0.05) from the control (Figure 3 ). Although a decrease in the lateral diameter of the lesions was also observed, these were not different from those of the control animals (P> 0.05). 2-Bromo-5- (2-methyl-2-nitrovinyl) -furan at this time did not show any effect (P> 0.05). Amphotericin B also showed no significant antileishmanial activity after 7 days of treatment, with an increase in the dorsal-ventral diameter similar to the control (P> 0.05). The lateral diameter of the lesions decreased slightly, but was not different (P> 0.05) from that observed in the control animals. At the end of the treatment (graph 4), the animals treated with 2-bromo-5- (2-bromo-2-nitrovinyl) -furan showed a decrease in the size of the lesions, which differed significantly with respect to the control, both in the dorsal-ventral diameter (P <0.05) as in the lateral (P <0.01). These variations were also statistically different (P <0.05) with respect to the animals treated with amphotericin B. However, in the animals treated with 2-bromo-5- (2-methyl-2-nitrovinyl) -furan it was not observed a significant effect.

Amphotericin B showed an increase in the dorso-ventral diameter similar to that of the control (P> 0.05). Although the lateral diameter of the lesions decreased slightly, it was not statistically different (P> 0.05) from the variation observed in the controls. In both graphs it can be seen that, although in the controls there is a greater growth of the dorsal-ventral diameter in relation to the lateral one, the effect of the products is more evident on the lateral diameter. In the present study it could be demonstrated that 2-bromo-5- (2-bromo-2-nitrovinyl) -furan has in vivo activity against Leishmania, according to the model of experimental cutaneous leishmaniasis developed in Balb / c mice inoculated with L. m . amazonensis 2-Bromo-5- (2-methyl-2-nitrovinyl) -furan, although it requires higher doses, also showed an effect in vivo against the experimental disease.

EXAMPLE 7 Treatment of human cutaneous leishmaniasis with 2-bromo-5- (2-bromo-2-nitrovinyl) -furan 100 patients with cutaneous leishmaniasis diagnosed parasitologically were treated. The treatment consisted of a daily application of an oil ointment base petrolatum containing 2-bromo-5- (2-bromo-2-nitrovinyl) -furan at 0.15% for 21 days.

In 90% of the patients a total regression of the lesions was reached, the healing time being dependent on the degree of progress that the disease had, so it ranged between 10 and 21 days. At the end of the treatment, the percentage indicated was not only the total regression of the ulcers but also the elimination of the local and systemic clinical symptoms that they presented, related both to the primary effect of the parasite and to secondary contamination by fungi and bacteria. There were no adverse reactions that prevented the continuation of the treatment.

Claims (6)

NOVELTY OF THE INVENTION CLAIMS

1. - The use of nitrovinylfuran derivatives for the preparation of a pharmaceutical composition useful for the treatment of leishmaniasis and trypanosomosis.

2. The use of nitrovinylfuran derivatives as claimed in claim 1, wherein the leishmaniasis comprises the cutaneous, mucocutaneous and visceral clinical forms and the trypanosomosis refers to that caused by Trypanosoma cruzi.

3. The use of nitrovinylfuran derivatives as claimed in claims 1 and 2, wherein the nitrovinylfuranic derivatives are: 2- (2-nitrovinyl) -furan, 2-bromo-5- (2-nitrovinyl) -furan, 2 - (2-bromo-2-nitrovinyl) -furan, 2-bromo-5- (2-bromo-2-nitrovinyl) -furan, 2- (2-methyl-2-nitrovinyl) -furan or 2-bromo-5- (2-methyl-2-nitrovinyl) -furan.

4. The use of nitrovinylfuran derivatives as claimed in claims 1 to 3, wherein the pharmaceutical composition is adapted to be administrable topically or systemically.

5. The use of nitrovinylfuran derivatives as claimed in claims 1 to 4, wherein the pharmaceutical composition contains oily ointments, oils and glycols, depending on the route of administration.

6. The use of nitrovinylphuran derivatives as claimed in claims 1 to 5, wherein the pharmaceutical composition is useful for the treatment of leishmaniasis and opportunistic bacterial and fungal diseases associated with lesions caused by leishmaniasis.