BR102016017666A2 - COMPOSITION AND USE OF A PEPTIDE - Google Patents

Abstract

The present invention relates to the composition comprising the peptide as defined in sequence: 1, 3, 4 or 5 and a pharmaceutically acceptable carrier. The present invention further relates to the use of a peptide as defined in sequence: 1, 3, 4 or 5 for the production of a pharmaceutical composition for the treatment of neglected diseases. The present invention further relates to a method of treating neglected diseases.

Description

(54) Title: COMPOSITION, E, USE OF A PEPTIDE (51) Int. Cl .: A61K 38/10; A61P 33/02 (73) Holder (s): BUTANTAN FOUNDATION (72) Inventor (s): MARIA CAROLINA QUARTIM BARBOSA ELIAS SABBAGA; CHRISTIANE BEZERRA DE ARAÚJO (74) Attorney (s): Dl BLASI, PARENTE & ASSOCIADOS ADVOGADOS (57) Abstract: The present invention relates to the composition comprising the peptide as defined in SEQ ID NO: 1, 3, 4 or 5 and one pharmaceutically acceptable vehicle. The present invention further relates to the use of a peptide as defined in SEQ ID NO: 1, 3, 4 or 5 for the production of a pharmaceutical composition for the treatment of neglected diseases. The present invention also relates to a method for treating neglected diseases.

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Invention Patent Descriptive Report for:

“COMPOSITION, AND, USE OF A PEPTIDE.

FIELD OF THE INVENTION [0001] The present invention relates to the composition comprising the peptide as defined in SEQ ID NO: 1, 3, or 5, and a pharmaceutically acceptable carrier. The present invention further relates to the use of a peptide as defined in SEQ ID NO: 1, 3, 4 or 5 for the production of a pharmaceutical composition for the treatment of neglected diseases. The present invention also relates to a method for treating neglected diseases.

BACKGROUND OF THE INVENTION [0002] The integrity of cellular processes depends on the appropriate balance of different proteins. One way to control the activity of such proteins is to control their half-life by proteolysis. In cells, there are two main pathways of destruction: lysosomal proteolysis or ubiquitin-proteasome pathway (Jayhyuk et al., 2001).

[0003] The ubiquitin-proteasome pathway is a process dependent on ubiquitin and ATP. The activation and inactivation of key proteins or protein complexes occurs through phosphorylation and dephosphorylation of proteins. The phosphorylation reactions that control the cell cycle are carried out by a specific set of protein kinases, whose activity increases and decreases over the cell cycle. THE

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2/20 activation and deactivation of these kinases at the right time is due to another set of proteins, called cyclins (Alberts et al. 2013).

[0004] Cyclines themselves do not show enzymatic activity, but when they bind to cell cycle kinases they become enzymatically active, which is why these kinases are known as cyclin-dependent protein kinases (Cdks). Cyclines are so called because their concentrations, unlike Cdks, vary cyclically during the cell cycle. This cyclic change assists in the periodic assembly and activation of the cyclin-Cdk complex. Once activated, the ciclinaCdk complex helps to trigger various cell cycle events, such as entry into the S or M phase (Alberts et al. 2013).

[0005] Intracellular peptides are constantly produced by the ubiquitin-proteasome system and probably many are functional. Peptide 5 (pep5 WELVVLGKL), from cyclin D2 G ₁ / S, showed a 2-fold increase during the S phase of the cell cycle of

HeLa. When fused to the cell penetrating peptide (cpp - YGRKKRRQRRR) it induced cell death in several tumor cells (Araujo et al., 2014).

[0006] The Trypanosomatidae family includes protozoa such as Trypanosoma cruzi, Trypanosoma brucei and Leishmania spp, agents that cause chagas disease,

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3/20 African sleep or trypanosomiasis, and leishmaniasis, respectively. According to the World Health Organization (WHO, 2013), these three diseases are listed among the 17 parasitic infections that affect people living in developing countries, known as neglected diseases.

[0007] Specifically, Chagas' disease represents one of the most significant public health problems in the American continent for the following reasons: number of infected people followed by death, socioeconomic impact and geographical distribution. Although the incidence of new infections has declined in Brazil and in other countries due to urbanization and improved living conditions, an estimated 8 million people remain infected (Bermudez et al., 2013).

[0008] Patent application BR 10 2014 000521 8, filed on January 9, 2014, from PROTEIMAX BIOTECNOLOGIA LTDA (BR / SP), entitled “peptide, pharmaceutical composition, in vitro and / or in vivo cell inhibition method in division, method of in vitro identification of cells in division and uses ”, describes the use of pep5-cpp, and its variants, for the production of a drug for the treatment of tumors. However, at no time in the application mentioned above, it is suggested to use pep5-cpp for the treatment of

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Wounds.

[0009] The present patent application presents a new use for the molecule pep5-cpp, capable of promoting effects on the proliferation and death of Chagas disease, sleep and leishmaniasis parasites. Neglected diseases are those caused by infectious agents or parasites and are considered endemic in low-income populations, with low investment in research. Diseases such as

Chagas disease, sleeping sickness and visceral leishmaniasis remain some of the main causes of morbidity and mortality worldwide. These diseases, known as neglected diseases, incapacitate or kill millions of people and represent an important medical need that remains unmet. Neglected diseases require the involvement of the entire society in order to gradually reduce the damage and mortality generated by these diseases.

SUMMARY OF THE INVENTION [0010] Considering the few treatment options for neglected diseases, especially for

Chagas, sleeping sickness and leishmaniasis, it is extremely important to study new drugs for these diseases. The drugs used to treat chagas disease, Nifurtimox and benznidazole, have several side effects, such as skin changes

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5/20 (photosensitive dermatitis and polymorphic erythema), gastric intolerance, myelosuppression and reversible dose-dependent peripheral neuritis, usually at the end of the treatment period (Viotti et al., 2009; Pinazo et al, 2013). In addition, they have limited effectiveness, especially in the chronic phase of the disease. In this case, a new product with antiparasitic action would be of great interest.

[0011] In one aspect, the present invention relates to the composition comprising the peptide as defined in SEQ ID

NO: 1, 3, 4 or 5, and a pharmaceutically acceptable carrier.

In one embodiment, said peptide is in a concentration from 1 μΜ to 50 μΜ. In another embodiment, said peptide is at a concentration of 25 μΜ.

[0012] In another aspect, the invention relates to the use of a peptide as defined in SEQ ID NO: 1, 3, 4 or 5 for the production of a pharmaceutical composition for the treatment of neglected diseases. In one embodiment, neglected diseases are selected from Chagas disease, sleeping sickness or leishmaniasis. In another embodiment, said peptide is in a concentration of 1 μΜ to 50 μΜ.

In another embodiment, said peptide is at a concentration of 25 μΜ.

[0013] In another aspect, the invention relates to a method for treating neglected diseases comprising administering a therapeutically effective amount of

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6/20 peptide as defined in SEQ ID NO: 1, 3, 4 or 5 or the composition as defined in any of claims 3 to a patient suffering from one of said disease. In one embodiment, neglected diseases are selected from Chagas disease, sleeping sickness or leishmaniasis.

BRIEF DESCRIPTION OF THE FIGURES [0014] The purpose of the invention, together with additional advantages of the same, can be better understood by reference to the attached figures and the following descriptions: [0015] Figure 1 shows the induction of death of T. cruzi CL Brener by pep5-cpp. 3x10 ⁶ parasites treated or not with pep5-cpp and analyzed by flow cytometry. A total of 10,000 events were analyzed in each group. NT: untreated; Cpp: carrier; Pep5-cpp: peptide 5 covalently linked to the carrier. The graph represents the mean and SEM of two experiments carried out in triplicates and independently.

[0016] Figure 2 shows epimastigote forms of Y in the presence of pep5-cpp. Parasites treated or not with pep5cpp and analyzed by flow cytometry. A total of 10,000 events were analyzed in each group. NT: untreated;

Pep5-cpp: peptide 5 covalently linked to the carrier. The graph represents the mean and SEM of two experiments carried out in triplicates and independently.

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7/20 [0017] Figure 3 shows the dose response of pep5-cpp in LLC-MK2 cells. 2x10 ⁵ cells were subjected to treatment with different concentrations of pep5-cpp for a period of 20 hours. After this time, the cells were trypsinized, marked with propidium iodide and analyzed by flow cytometry. A total of 10,000 events were analyzed per group. The graph represents the mean and SEM of two experiments carried out in triplicates and independently.

[0018] Figure 4 shows that pep5-cpp changes the number of cells infected with T. cruzi. LLC-MK2 cells were cultured in 12-well plates (2x10 ⁴ / well) and after 1h infected with 1x10 ⁶ parasites (T. cruzi strain Y), in the presence or not of pep5-cpp. 20 hours after infection, the medium was replaced with complete new medium and the cells were maintained in culture for an additional 24 hours. After this period, the cells were fixed in 4% PFA, stained with

Giemsa and analyzed by MOC (AT: 1000X). A total of 100 cells were analyzed per group. A: infected and untreated cells; B: cells infected and treated with pep5cpp (simultaneously). The arrows point to intracellular parasites, amastigote shape. The graph represents the mean and SEM of two experiments carried out in duplicates and independently.

[0019] Figure 5 shows LLCMK2 cells that were

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8/20 infected and treated with pep5-cpp for approximately 20h and then the medium was replaced with pep5-cpp free medium. The number of trypomastigotes was counted in a Neubauer chamber on days 4, 5, 6 and 7 after infection. As can be seen in the graph, the number of parasites in the supernatant decreased dramatically in the group treated with pep5-cpp.

[0020] Figure 6 shows that pep5-cpp changes the distribution of cells throughout the cell cycle in T.

brucei. 1x10 ⁶ cells were treated with pep5-cpp for a period of 3 hours. After this time, the cells were trypsinized, marked with propidium iodide and analyzed by flow cytometry. A total of

10,000 events were analyzed per group. The graph represents the mean and SEM of two experiments carried out in triplicates and independently.

[0021] Figure 7 shows that pep5-cpp causes arrest in

L. amazonensis. 1x10 ⁶ cells were treated with pep5-cpp for a period of 3 hours. After this time, the cells were trypsinized, marked with propidium iodide and analyzed by flow cytometry. A total of 10,000 events were analyzed per group. The graph represents the mean and SEM of two experiments carried out in triplicates and independently.

[0022] Figure 8 shows the effect of pep5-cpp 6

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9/20 amino acids. The parasites were treated with pep5-cpp

6aa (SEQ ID NO: 3) (25μM) in LIT medium for approximately 3 hours and then analyzed by flow cytometry. The graph shows the percentage of cells in each phase of the cell cycle, as well as the number of cell deaths for each group. Pep5 alone (WELVVLGKL), that is, without the carrier, has no significant effect on the proliferation and cell death of the parasites, while the smallest active form, pep5cpp 6aa (WELVVL-cpp - SEQ ID NO: 3), has similar effects to the original form of pep5-cpp (WELVVLGKL-cpp). DETAILED DESCRIPTION OF THE INVENTION [0023] Although the present invention may have different embodiments, it is shown in the drawings and in the following detailed discussion, a preferred embodiment with the understanding that the present description should be considered an example of the principles of the invention and is not intended to limit the present invention to what has been illustrated and described here.

DEFINITIONS [0024] Unless otherwise defined, all scientific and technical terms are understood to have the same meanings commonly used in the technique to which they belong. For the purpose of this disclosure, the following terms are defined.

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10/20 [0025] The terms trypomastigote, epimastigote and amastigote must be defined as follows:

[0026] - trypomastigote: the kinetoplast is close to the posterior end of the body, and the flagellum posterior to the nucleus, connected by a wide wavy membrane.

Metacyclic trypomastigote have an elongated nucleus with the flagellum emerging from the posterior end of the cell associated with the spherical kinetoplast;

[0027] - epimastigote: the flagellum leaves the cell anterior to the nucleus. The kinetoplast is located between the nucleus and the anterior end; and [0028] - amastigote: the flagellum is extremely short as it protrudes from the flagellar pouch.

[0029] The term cell-penetrating peptide or CPP refers to short amino acid sequences that facilitate the entry of the main peptide into the intracellular medium and thus acts as a mode of transporting peptides to the intracellular medium. The preferred CPP in this application is defined in SEQ ID NO: 2 - YGRKKRRQRRR. Still in the context of the present patent application, hydrophilic tails known in the prior art, such as the TAT, SynB1, SynB3, PTD-4, PTD-5, DTat, R9-Tat, among others, are included in the CPPs . Amphiphilic tails are also included, such as MAP, SBP, FBP tails, among others.

[0030] The term pep5-cpp refers to the peptide pep5

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11/20 bound to the cpp peptide at the C-terminal portion. The pep5-cpp sequence, referred to in the present application as SEQ ID NO: 1, is WELVVLGKLYGRKKRRQRRR. The present application also comprises deletions of amino acids in the N-terminal portion of pep5,

represented by the strings SEQ ID AT THE: 3 (pep5-cpp 3 - deletion of three amino acids), SEQ ID AT THE: 4 (pep5-cpp 2 - deletion of two amino acids) and SEQ ID AT THE: 5 (pep5-cpp 1 - deletion in one amino acid), connected The peptide tail represented by SEQ ID NO: 2. [0031] O treatment term refers to if The a method for

relieve or cancel a disease and / or its concomitant symptoms.

[0032] The term therapeutically effective amount refers to the sufficient amount of the compound to be administered to prevent the development, or to relieve to some extent, one or more of the symptoms of the condition or disorder to be treated.

[0033] The term "pharmaceutically acceptable carrier" refers to a non-toxic, inert solid, semi-solid liquid excipient, diluent, auxiliary formulation of any kind, such as saline, and water. Some examples of materials that can serve as pharmaceutically acceptable carriers are sugars, such as lactose, glucose and sucrose, starches, such as corn starch and potato starch, cellulose and its derivatives,

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12/20 such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate, cyclodextrin; oils, such as peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol, polyols, such as glycerin glycol, sorbitol, mannitol and polyethylene; esters, such as ethyl laurate, ethyl oleate, agar; buffering agents, such as aluminum hydroxide and magnesium hydroxide; alginic acid; isotonic saline, Ringer's solution; buffer solutions of ethyl alcohol and phosphate, oily water emulsion containing mycobacteria killed by the action of heat or components of its cell wall (complete adjuvant of

Freund), as well as other compatible non-toxic substances used in pharmaceutical formulations.

METHODOLOGY [0034] Peptide Synthesis: Peptide 5 and its variants (SEQ ID NO: 1, 3, 4 and 5) were synthesized coupled to the cell penetrating peptide-sequence YGRKKRRQRRR, TAT 47-57) , covalently linked to the C terminal of the peptide mentioned above. In all assays, peptides> 95% pure were used, with pep5-cpp being supplied by

Proteimax Biotecnologia LTDA, São Paulo, SP, Brazil. The peptide was diluted in MilliQ water and stored at -20 ° C

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13/20 until the time of the experiments. The tests were carried out in concentrations ranging from 1 to 50 μΜ, and the compound was added to the culture medium of the cells and / or parasites subjected to the treatment.

[0035] Cell culture: epimastigote forms of

Trypanosoma cruzi, strains Y and CL Brener, were cultured in LIT (Liver Infusion Triptose) medium containing 10% fetal bovine serum (SFB) at 28 ° C, in a concentration of 3x10 ⁶ parasites / ml. Leishmania amazonensis was cultivated in 199 (Gibco) medium and the procyclic forms of

Trypanosoma brucei were maintained in SDM-79 medium, 10%

SFB, 28 ° C. LLC-MK2 mammalian cells (Monkey Kidney

Cell) were maintained in DMEM, 10% SFB at 37 ° C, 5% CO ₂ .

[0036] Infection assay: Monkey kidney epithelial cells (LLC-MK2) were maintained in DMEM 10% fetal bovine serum, 37 ° C and 5% CO2 throughout the infection assay. The parasites (T. cruzi cepa Y) were added to the cell culture and left for a period of 24 hours, in the presence or not of pep5-cpp. During the treatment with pep5-cpp the cells were maintained in DMEM without SFB. After treatment, the cells were fixed in 4% paraformaldehyde, dehydrated in xylol-acetone baths and stained with

Eosin and Methylene Blue. The slides were analyzed using standard light microscopy and the number of infected cells, as well as the number of intracellular parasites, was counted

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14/20 in both the control and experimental groups.

[0037] Flow cytometry: Flow cytometry analyzes were performed on an Attune Acoustic Focusing device

Cytometer (Applied Biosystems). For cell cycle analysis, the parasites were washed with PBS and centrifuged at 2500 rpm, 5 minutes. The samples were fixed in ethanol

70% in PBS for at least 1 hour on ice and then centrifuged again (2500 rpm, 5 min). The pellet was resuspended in PBS containing propidium iodide (PI; 1 mg / mL), RNAse (10mg / mL) and maintained for 30 minutes on ice.

A total of 10,000 events were analyzed per sample. COMPOSITION [0038] In a first embodiment, the present invention relates to a composition comprising the peptide as defined in SEQ ID NO: 1, 3, 4 or 5 and a pharmaceutically acceptable carrier. In a further embodiment, said peptide is in a concentration of μΜ to 50 μΜ. In another additional embodiment, said peptide is at a concentration of 25 μΜ.

USE OF A PEPTIDE [0039] In another embodiment, the present invention also relates to the use of a peptide as defined in SEQ

ID NO: 1, 3, 4 or 5 for the production of a pharmaceutical composition for the treatment of neglected diseases.

In a further embodiment, neglected diseases

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15/20 are selected from Chagas disease, sleeping sickness or African trypanosomiasis and leishmaniasis. In another additional embodiment, said peptide is in the composition at a concentration of 1 μΜ to 50 μΜ. In another additional embodiment, said peptide is in the composition at a concentration of 25 μΜ.

METHOD FOR TREATING NEGLECTED DISEASES [0040] In another embodiment, the present invention also relates to a method for treating neglected diseases, comprising administering a therapeutically effective amount of the peptide as defined in SEQ ID

NO: 1, 3, 4 or 5 or the composition as defined in any one of claims 1 to 3 to a patient suffering from said diseases. In an additional embodiment, neglected diseases are selected from

Chagas, sleeping sickness or African trypanosomiasis and leishmaniasis

EXAMPLES

EXAMPLE 1 - Effect of pep5-cpp on the epimastigote form of T.

cruzi [0041] The life cycle of T. cruzi alternates in non-replicative, but infectious (trypomastigote and metacyclic trypomastigote) and replicative, but not infective (epimastigote and amastigote) stages. The epimastigote forms of T. cruzi (strains CL Brener and Y) were

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16/20 treated with different concentrations of cpp (cell penetrating peptide) or with pep5-cpp for 3 hours. The cells were fixed in 70% ethanol in PBS (overnight), incubated with RNAse (10mg / mL), propidium iodide (1mg / mL) and analyzed by flow cytometry. Figure 1 shows the percentage of dead cells in the CL Brener strain, and as noted, the effect of the peptide starts from the concentration of 10 μΜ.

[0042] Based on the effect caused on the CL strain

Brener, another experiment was performed with strain Y, commonly used by our group as a model of infection in mammalian cells. In figure 2, it can be seen that at a concentration of 25 μΜ, about 60% of the parasites are dead, when compared to the control group (only 20% of dead parasites). Thus, we can conclude that pep5-cpp had a trypanocidal effect on the two strains of T. cruzi used in this work.

EXAMPLE 2 - Pep5-cpp dose-response assay in cells

LLC-MK2 [0043] LLC-MK2 (monkey kidney cell) cells are fibroblasts widely used as an in vitro infection model for T. cruzi. Thus, a study on the sensitivity of these cells to the effect of this fragment of cyclin was carried out. As shown in figure 3,

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17/20 LLC-MK2 cells demonstrated high resistance to the cell death-inducing action promoted by pep5-cpp. Only at the highest concentration used (50 μΜ, 20 hours of treatment) was pep5-cpp able to induce about 25% of cell death.

[0044] Thus, based on the results presented above, the effect of pep5-cpp on infection caused by T. cruzi was verified, as well as its effect on the replication of the parasite within host cells. For this purpose, concentrations between 10 and 25 μΜ were used, which only promoted the death of the protozoa, but not the host cells.

EXAMPLE 3 - Pep5-cpp in in vitro infection of T. cruzi [0045] LLC-MK2 cells were cultured in 12-well plates (2x104 / well) and after 1h infected with 1x10 ⁶ parasites (T. cruzi trypomastigote, strain Y ), in the presence or not of pep5-cpp, 20 hours after infection, the medium was replaced with a complete new medium and the cells were kept in culture for an additional 24 hours. After this period, the cells were fixed in 4% PFA, stained with

Giemsa and analyzed by MOC (AT: 1000X).

[0046] As seen in figure 4, treatment with pep5-cpp (25μΜ, 20 hours) during infection, decreased the number of infected cells, as well as the amount of parasites in the intracellular medium.

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EXAMPLE 4 - Pep5-cpp and its action on T. brucei and L.

amazonensis [0047] The effect of pep5-cpp on the other trypanosomatids was also evaluated. The tests were performed based on the data obtained in T. cruzi. The procyclical form of the causative agent of sleeping sickness, T.

brucei, strain 927, was treated with pep5-cpp for 3 hours.

Flow cytometry analyzes demonstrated a different effect than that observed for T. cruzi and the other mammalian strains treated with this intracellular peptide. In figure 5, it can be seen that cells in the presence of pep5-cpp, even in high concentrations, tend to have a different distribution from the control throughout the phases of the cell cycle. At a concentration of 50 μΜ, the largest percentage of cells are with

DNA greater than 4N (hyperploidy). However, it was not possible to observe a significant increase in cell death.

[0048] The promastigote form of Leishmania amazonensis, when treated with pep5-cpp, undergoes arrest in the S phase of the cell cycle in both concentrations (Figure 6).

[0049] The different species of trypanosomatids behave differently in the presence of pep5-cpp, causing death in T. cruzi and arrest in the other two organisms used in this work.

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EXAMPLE 5

Effect of pep5-cpp 6 amino acids (SEQ ID NO: 3) on the epimastigote form of T. cruzi CL Brener.

[0050] The parasites were treated with pep5-cpp 6aa (SEQ ID NO: 3) (25μΜ) in LIT medium for approximately hours and then analyzed by flow cytometry.

Figure 8 shows the percentage of cells in each phase of the cell cycle, as well as the number of cell deaths for each group. The smallest active form pep5-cpp 6aa (WELVVL-cpp) (SEQ ID NO: 3) has similar effects to the original form of pep5-cpp (WELVVLGKL-cpp) (SEQ ID NO: 1). Pep5 alone (WELVVLGKL), that is, without the carrier, has no significant effect on the proliferation and cell death of the parasites.

[0051] In view of the result above, we understand that the other active forms of pep5 (WELVVLGK-cpp and WELVVLGcpp) should have a similar function in the parasites used in this study.

REFERENCES [0052] Alberts, B .; Bray, D .; Hopkin, K .; Johnson, A .;

Lewis, J .; Raff, M .; Roberts, K .; Walter, P. Essential

Cell Biology, Fourth Edition, 2013, 838 p.

[0053] Araujo, C.B .; Russo, L.C .; Castro, L.M .; Forti,

F.L .; Monte, E.R .; Rioli, V .; Gozzo, F.C .; Colquhoun, A;

Ferro, E.S. A novel intracellular peptide derived from

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Gi / S cyclin D2 induces cell death. The Journal of Biological Chemistry, v. 289, No. 24, p. 16711-16726, 2014. [0054] Bermudez, J .; Davies, C .; Simonazzi, A .; Real,

J.P .; Palma, S. Current drug therapy and pharmaceutical challenges for Chagas disease. Acta Tropica, v. 156, p. 116, 2016.

[0055] Jayhyuk, M .; Kyung, B.K .; Craig, M. The

Ubiquitin-Proteasome Pathway and Proteasome Inhibitors.

Med Res Rev., 21 (4), p. 245-273, 2001.

[0056] Pinazo, M.J .; Guerrero, L .; Posada, E .;

Rodriguez, E .; Soy, D; Gascon, J. Benznidazole-Related

Adverse Drug Reactions and Their Relationship to Serum Drug

Concentrations in Patients with Chronic Chagas

Disease. Antimicrob. Chemother Agents. 57 (1), p. 390-395,

2013.

[0057] Viotti, R,; Vigliano, C .; Lococo, B .; Alvarez,

M.G .; Petti, M .; Bertochi, G .; Armenti, A. Side effects of benznidazole as treatment in chronic Chagas disease: fears

and realities. 163, 2009. Expert Rev. Anti-Infect. The R • 7, p. 157- [0058] Who, 2013. Sustaining the drive to overcome the global impact of neglected tropical diseases : second who

report on neglected diseases.

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Claims (7)

1. Composition characterized by the fact that it comprises the peptide as defined in SEQ ID NO: 1, 3, 4 or 5 and a pharmaceutically acceptable carrier. 2. Composition, according to claim 1, characterized by the fact that said peptide is in a concentration of 1 μΜ to 50 μΜ. 3. Composition, according to claim 2, characterized by the fact that said peptide is in a concentration of 25 μΜ. 4. Use of a peptide as defined in SEQ ID NO: 1, 3, 4 or 5, characterized by the fact that it is for the production of a pharmaceutical composition for the treatment of neglected diseases. 5. Use, according to claim 4, characterized by the fact that neglected diseases are selected from Chagas disease, sleeping sickness or leishmaniasis. 6. Use, according to claim 4 or 5, characterized by the fact that the peptide is in the composition in a concentration of 1 μΜ to 50 μΜ. 7. Use, according to claim 6, characterized by the fact that the peptide is in the composition at a concentration of 25 μΜ. Petition 870160040354, of 7/29/2016, p. 52/62 1/8 100η ρ <0.05 * ρ <0.001 ΝΤ CPP 50μΜ Pep5-cpp 50μΜ