DISEASES IN HUMAN OR ANIMAL

[001] The present invention refers to the field of inflammation and treatment and/or prevention of acute or chronic inflammatory pathologies, which may be associated with fibrotic processes for human and/or animal use.

[002] The present invention is directed to the use of diethylcarbamazine (DEC) , and/or its pharmaceutically acceptable salts, incorporated to pharmaceutical and veterinary compositions, in the manufacture of a medicament for the treatment or prevention of acute and/or chronic inflammatory pathologies, which may be associated with fibrotic processes in human beings and animals. The present invention also refers to a pharmaceutical composition containing an effective amount of DEC, and/or its pharmaceutically acceptable salts, and excipients.

[003] The present invention was developed mainly for the treatment and/or prevention of acute or chronic human and veterinary inflammatory diseases associated or not with fibrotic processes, in particular, but not exclusively, or limited to: chronic obstructive disease, asthma, pulmonary hypertension, acute pulmonary inflammation such as respiratory distress syndrome (ARDS), pleuritis, hepatitis, cirrhosis, asthma, arthritis, arthrosis, systemic sclerosis, sclerodermatous graft-versus-host disease in bone marrow transplant recipients, Crohn's disease, cystic fibrosis, pulmonary fibrosis, intestinal fibrosis, liver fibrosis, kidney fibrosis, endomyocardial fibrosis, retroperitoneal fibrosis, and keloid.

[004] The present invention is based on experiments demonstrating that DEC, and/or its pharmaceutically acceptable salts, incorporated into a pharmaceutical and/or veterinary composition (generally called NANO-DEC) when administered via one of the administration routes described below, exhibits greater anti ^¬ inflammatory and anti-fibrotic response than DEC administered alone.

[005] To enhance the effectiveness, the NANO-DEC may be administered or combined with one or more anti-inflammatory, or immunomodulatory, corticoid, analgesic, anti-fibrotic, antitumor, antimycobacterial , fungicidal or antibiotic or immunobiological agents commonly indicated or used for the treatment or prophylaxis of inflammatory and fibrosis diseases.

[006] The use of such pharmaceutical and/or veterinary compositions containing DEC, and/or its pharmaceutically acceptable salts, may help prevent or treat various acute or chronic

inflammatory and fibrotic conditions. As described in the use of pharmaceutical or veterinary compositions as a filaricidal agent, in the treatment of microfilariae infections.

[007] Inflammation is "the body's immediate response to damage to its tissues and cells by pathogens, noxious stimuli such as chemicals, or physical injury" [Weiss, 2008].

[008] According to the persistence time of the inflammatory process, the inflammation can be of two types: acute or chronic. Acute inflammation is a short-term response that usually results in healing: polymorphonuclear leukocytes (neutrophils) infiltrate the damaged region, removing the stimulus and repairing the tissue. Chronic inflammation, by contrast, is a prolonged, dysregulated and maladaptive response that involves active inflammation, tissue destruction and attempts at tissue repair. The main characteristic cells in chronic inflammation are the mononuclear leukocytes (lymphocyte and macrophage) . This inflammation (chronic) is usually associated with many chronic human diseases, and diseases, including cancer, allergies, atherosclerosis, arthritis, autoimmune diseases, among others [Weiss, 2008].

[009] Despite the existence of several drugs for the treatment of inflammations caused by different etiologies, such as nonsteroidal anti-inflammatory drugs (NSAIDs) and steroids. The therapeutic options, in special for chronic process are very limited, with limited efficacy, high incidence of adverse effects, and almost never promote a definitive solution to the disease.

[010] Fibrosis is a pathological process characterized by excessive accumulation of connective tissue components in an organ or tissue as part of a healing process or fibroid degeneration. Fibrogenesis is a multistage process, now increasingly seen as the result of deregulated tissue repair responses, that occurs in response to chronic tissue injury and/or chronic inflammation, in which aberrantly sustained production of cytokines, growth factors, and angiogenic factors tilt tissue homeostasis towards interstitial hyperplasia and excessive accumulation of extracellular matrix. Progressive fibrosis, which disrupts the normal tissue architecture and results in progressive loss of organs functions, which ultimately leads to their dysfunction and failure, is recognized to be one of the major causes of morbidity and mortality in individuals [Rosenbloom et al . , 2010; Ho et al., 2014] .

[011] Fibrotic diseases affect a wide spectrum of organs and a large number of persons, and their devastating effects cause an enormous burden on health resources with severe economic

consequences. The usually progressive nature of these diseases and the absence of effective treatment drugs, or therapies that are capable to reverse existing fibrotic lesion are not yet available, and increase the seriousness of the problem [Rosenbloom et al . , 2010; Ho et al. , 2014] .

[012] The fibrotic diseases encompass a broad spectrum of clinical diseases, including multisystemic diseases, such as systemic sclerosis, multifocal fibrosclerosis , Crohn's disease, sclerodermatous graft-versus-host disease in bone marrow transplant recipients, as well as organ-specific disorders, such as pulmonary, liver, intestine, myocardium, peritoneum and kidney fibrosis. [013] Inflammatory and fibrotic diseases are commonly associated with high morbidity and mortality. Besides, conventional drug treatments (in particular through the use of nonsteroidal anti ^¬ inflammatory drugs (NSAIDs) and steroids have low efficacy and serious adverse effects. It is clear the need to research new, more effective and safe treatments to solve the problem of inflammation, especially the chronic inflammatory processes, which are often associated with fibrosis.

[014] In this context, the present invention discloses the use of pharmaceutical and/or veterinary compositions containing diethylcarbamazine (DEC) as a safe anti-inflammatory and anti- fibrotic, for use in the treatment or prophylaxis of acute and/or chronic inflammatory conditions, associated or not to fibrotic processes, for human or animal uses.

[015] Dietilcarbamazine, has as chemical name 1- diethylcarbamyl-4-methylpiperazine, its CAS Registry Number is 90- 89-1. Its molecular formula is C10H21N3O, it molecular weight is 199,293 g/mol, and its structural formula is presentd below in Figure 1.

[016] Diethylcarbamazine is a widely used drug in the treatment of filariasis and has excellent tolerability with low side effects.

Commercially the DEC is sold as salts, where the most common are hydrochloride, citrate or phosphate.

[017] Although millions of doses are distributed free of charge against filariasis worldwide, the use of DEC generates some adverse reactions. The most commonly are nausea, vomiting, abdominal pain, diarrhea, and headaches [Francis et al . , 1985; Patrono et al . , 1981] .

[018] Since 1950, DEC has been marketed in the form of citrated salt by numerous pharmaceutical companies under different names. It is a white powder, very water soluble, stable, even in conditions of very high humidity and temperature, and also resisting autoclaving The name DEC generally refers to its citrated form, since it is most commonly used [Dreyer & Noroes, 1997] .

[019] DEC is rapidly absorbed from the gastrointestinal tract, reaching a peak plasma concentration between one and three hours after oral intake, not concentrating on any specific organ, being metabolized in the liver and its excretion is basically renal [Ilondu et al. , 2000] .

[020] Part of the filaricidal effects attributed to DEC is due its interference in the arachidonic acid metabolism. This change in the arachidonic acid metabolism gives to DEC its anti-inflammatory properties [Maizel & Denham, 1992; Peixoto & Silva, 2014] .

[021] The arachidonic acid pathway includes the lipoxygenase (LOX) and cyclooxygenase (COX) enzymes. The COX pathway exhibits similarity to the nitric oxide pathway, both of which have constitutive and inducible isoforms of their enzymes and control inflammatory responses [McGarry et al . , 2005].

[022] Some clinical reports have shown good results with the use of DEC in patients with bronchial asthma, where the treatment was effective in containing the acute attacks caused by the disease [Srinivas & Antani, 1971; Liu et al . , 1997].

[023] Queto et al . (2010) showed that DEC has an important action in blocking pulmonary eosinophilic inflammation in mice sensitized with ovalbumin. According these authors DEC blocks pulmonary hyperreactivity, cytokine production and eosinophilia in vivo and in vitro.

[024] Similar results were obtained using another experimental model, where Ribeiro et al . (2017) evaluated the anti-inflammatory activity of DEC in a model of acute pulmonary inflammation ( carrageenan-induced pleurisy) . Injection of carrageenan into the pleural cavity induced accumulation of fluid containing a large number of polymorphonuclear cells (PMNs), as well as infiltration of PMNs into lung tissue and enhaced the expression of nitrite, tumor necrosis factor a (TNF- ) , expression of interleukin-ΐβ (IL- 1β), cyclooxygenase (COX-2), inducible nitric oxide synthase (iNOS) and nuclear factor kappa B (NF-κΒ) .

[025] Following oral administration of DEC (50 mg / kg), a significant reduction occurred in all markers of inflammation. The authors demonstrated that DEC treatment significantly decreased COX- 2 expression in lung tissue as observed with other non-steroidal anti-inflammatory drugs [Ribeiro et al . , 2014]. Moreover, in addition to inhibiting lipoxygenases (LOX) and cyclooxygenase (COX) , DEC also blocks the activation of NF-κΒ. Experimental evidence clearly suggests that NF-κΒ plays a central role in regulating several genes responsible for the generation of mediators or proteins in lung inflammation, as well as hepatic inflammation such as TNF- , IL-Ιβ, iNOS and

COX-2 [ Impellizzeri et al . , 2011] . Therefore, inhibition of the release of TNF-a IL-a 1β by DEC could be attributed to the inhibitory effects of NF-κΒ activation.

[026] Other studies have analyzed the potential of DEC against alcoholic hepatitis, conferring a decrease in the activity of transaminases, pro-inflammatory cytokines (TNF-a and IL-6) and nuclear kappa B factor [Santos Rocha et al . , 2012] .

[027] Rodrigues et al . (2015) demonstrated that treatment with DEC was able to reduce liver damage, collagen content, expression of inflammatory markers; (total cholesterol, high-density lipoprotein cholesterol, triglyceride content and aspartate aminotransferase) , as well as increasing the expression of anti ^¬ inflammatory markers (AMPK and interleukin-10) .

[028] In experimental design of chronic liver disease induced by carbon tetrachloride (CCL4) , the hepatocellular damage observed by the presence of necrosis, fibrosis and inflammatory infiltrates was reduced in the animals treated with DEC. [029] Fibrotic areas and inflammatory markers such as COX-2, IL-Ιβ, MDA, TGF-β and aSMA were also significantly decreased after DEC treatment. On the other hand, DEC increased the expression of the anti-inflammatory cytokine IL-10, thus conferring an anti- inflammatory action. In this work, it was concluded that DEC presentd benefits acting as a hepatoprotective, anti-inflammatory and anti- fibrotic drug [Rocha et al . , 2014] .

[030] Different animal models and experimental approaches have also indicated the use of DEC as a potential immunomodulator . U.S. Patent 4,900,548A claims the use of DEC as a stimulator of antigen- antibody immunological interactions with possible antiviral action. U.S. patent 4578268A further claims subcutaneous and intramuscular implants of slow release DEC compositions for controlling filariasis over a long period of time.

[031] The only study that have been reported regarding the incorporation of DEC in pharmaceutical compositions is the work of Siram et al . , 2014, where solid lipid nanoparticles of DEC citrate were obtained for treatment of filariasis in order to change pharmacokinetic properties, in particular the retention time of the drug in the body. The authors observed that the incorporation of DEC into the nanoparticles increase the amount of DEC that reaches the lymphnodes and the retention time of the drug in the body, thus generating a better therapeutic effect. These reports, show that DEC has proven anti-inflammatory, immunomodulatory and anti-fibrotic properties, in addition to its main therapeutic use as a filaricidal drug .

[032] In the last decades, the pharmaceutical industry has opted for the search of second uses of commercially available drugs. This procedure presents less cost than the process of discovering a new chemical entity (new drug) , and decrease the time required for approval of the use of the drug by regulatory agencies, due to less need for pre-clinical and clinical trials and tests. [033] In this way the present invention has its innovative character in claiming the use of pharmaceutical and/or veterinary compositions containing DEC, or its pharmaceutically acceptable salts, for therapeutic purposes for the treatment or prophylaxis of acute and / or chronic inflammatory conditions, associated or not with fibrotic processes.

[034] The present invention demonstrates that the incorporation of DEC, and/or its pharmaceutically acceptable salts, in pharmaceutical and veterinary compositions exhibits enhanced anti ^¬ inflammatory and anti-fibrotic, dose-reducing and treatment-time efficacy in experimental models of inflammation as compared to groups treated with DEC alone.

[035] It stands out that the incorporation of DEC, or its pharmaceutically acceptable salts, in nanosystems and/or other drug delivery systems allows the creation of a in situ controlled release system, capable to regulate and potentiate the anti-inflammatory and anti-fibrotic actions of DEC.

[036] The present invention also includes the use of any physiologically and/or pharmaceutically acceptable salts and/or any conventional base salts of DEC that are incorporated into pharmaceutical and/or veterinary compositions for use for therapeutic purposes for the treatment or prophylaxis of acute or chronic inflammatory conditions, associated or not to fibrosis. Examples of salts are selected from the group, but not limited to: alkali metal salts, preferably sodium and potassium salts, alkaline earth metal salts (calcium and magnesium salts), chloride, bromide, hydrochloride, acetates, maleate, mesylate, nitrate, tartrate, gluconate, citrates, carbonates, carbamates, phosphates, diphosphates, phosphonates , glycosides, sulfates, sulfonates and ammonium salts derived from amine or organic amines having from 1 to 16 carbon atoms. [037] According to the present invention, the administration of pharmaceutical and/or veterinary compositions containing DEC, can act systemically and/or locally, and may be performed by the following administration routes: oral, sublingual, nasal, rectal, intragingival , endovenous, intramuscular, intraarticular, subcutaneous, cutaneous (patch or transdermal patch) , inhalatory, transdermal, topical or spinal (subarachnoid and peridural), parenteral, con unctival, optic, or as an implant or stent, not being limited to these. In particular, the oral route is preferred.

[038] The suitable forms for oral administration are those which function according to the prior art, which release the pharmaceutical compositions containing DEC, or its pharmaceutically acceptable salts, rapidly and/or in a modified/controlled manner and which contain the DEC, or its pharmaceutically acceptable salts, according to the present invention in crystalline and/or amorphous forms and/or dissolved form or in the form of salts, for example tablets (tablets uncoated or coated, for example with resistance to gastric juice or by dissolution or insoluble coatings that control the release of the compound according to the present invention) , tablets or films/wafers, which disintegrate rapidly into the oral cavity, films/lyophilisates, capsules (for example hard or soft gelatine capsules), sugar coated tablets, granules, lozenges, powders, emulsions, suspensions , pills, powders, granules, powders, aerosols or solutions.

[039] Parenteral administration may bypass an absorption step

(for example, intravenously, intraarterially, intracardially, intraspinally or intralombally) or include an uptake (e.g., intramuscularly, subcutaneously, intracutaneously, percutaneously or intraperitoneal route) . Forms of administration suitable for parenteral administration include the injection and infusion preparations in the form of solutions, suspensions, emulsions, lyophilizates and sterile powders. [040] For the other administration route, suitable examples are inhalation medications (including powder inhalers, nebulizers), nasal drops, solutions or sprays; tablets for oral, sublingual or buccal administration, films or capsules/cachets, suppositories, ear or eye preparations, vaginal capsules, ova, aqueous suspensions (lotions, stirring mixtures), gels, lipophilic suspensions, ointments, creams, ointments, therapeutic systems transdermal (eg. adhesives), milk, pastes, foams, powders, implants or stents.

[041] The present invention provides pharmaceutical forms comprising pharmaceutical and/or veterinary compositions containing DEC, or its pharmaceutically acceptable salts, usually together with one or more pharmaceutically suitable auxiliaries (pharmaceutical ad uvants/excipients ) , and the use thereof for the purposes previously mentioned.

[042] The DEC-containing and/or DEC salts-containing compositions according to the present invention for generating a pharmaceutical or veterinary composition is carried out in a manner known per se by converting the DEC or DEC-salts to the desired form of administration with the usual pharmaceutically acceptable excipients for pharmaceutical compositions. Suitable carriers and compositions are known to those skilled in the art or may be obtained from, for example, United States Pharmacopoeia, European Pharmacopoeia, British Pharmacopoeia, Brazilian Pharmacopoeia, Remington's Pharmaceutical Sciences, 18th ed. (1990) .

[043] The pharmaceutically acceptable excipients used may, but not exclusively, or limited to: carrier substances, fillers, disintegrants , binders, wetting agents, absorbents and adsorbents, diluents, solvents, cosolvents, emulsifiers, solubilizers, flavoring agents, colorants, preservatives, stabilizers, wetting agents, salts for modifying osmotic pressure or buffers. Also considered as auxiliary agents are lipids, cyclodextrins , nonionic surfactants, polymers such as (acrylates and derivatives thereof, poly-ε- caprolactone, poly (lactic acid), polyglycolic acid, co-glycolic poly lactic acid, methacrylated derivatives, cholesterol, among others .

[044] The pharmaceutically acceptable excipients used in the context of the present invention may for example be, without being limited to: saccharides salts (mono-, di-, tri-, oligo- and / or polysaccharides), proteins, amino acids, peptides, fats, waxes, oils, hydrocarbons and the derivatives thereof and the auxiliaries may be of natural or synthetic or partially synthetic origin.

[045] The pharmaceutical and veterinary compositions of the present invention may be in solid form, for example, without being limited to: tablets, lozenges, pellets, suppositories, capsules, pills, powders, granules, films, cachets, sugar-coated or semi-solid transdermal systems, for example, as ointments, creams, gels, suppositories, emulsions or in liquid form, for example as solutions, tinctures, suspensions or emulsions. The pharmaceutical compositions of this invention may further be in the form of novel pharmaceutical forms including: liposomes, cyclodextrin complexes, nanospheres, nanocapsules , nanoparticles , microcapsules, microspheres, nanoemulsions , microemulsions and solid lipid particles.

[046] The present invention further provides the use of DEC, or its pharmaceutically acceptable salts, in pharmaceutical and/or veterinary compositions according to the present invention for the preparation of a therapeutic scheme for the treatment and/or prevention of acute and/or chronic inflammatory diseases, which may be associated with fibrotic processes, in humans and/or animals.

[047] The pharmaceutical and/or veterinary compositions of DEC, and its pharmaceutically acceptable salts, according to the present invention may be employed alone, individually or, if necessary, in combination with one or more other pharmacologically active substances, provided that this combination does not lead to unacceptable side effects. Accordingly, the present invention further provides a pharmaceutical and/or veterinary composition of DEC, and its pharmaceutically acceptable salts, according to the present invention and one or more other active compounds or medicaments and/or drugs, in particular used in the prophylaxis and/or therapy of inflammatory diseases associated or not with fibrotic processes.

[048] To the present, there are no technologies, or equivalent patents that can be compared with the present invention in essence. There are several active ingredients (diclofenac, ibuprofen, acetylsalicylic acid, etc.) and medicines and/or pharmaceutical forms (gels, tablets, capsules, injectables, etc.) that are useful and also used for inflammatory and fibrosis conditions, but there are no effective drugs to date with complete reversal various inflammatory and fibrotic processes, especially in chronic stage, this invention may become a new alternative.

[049] The present invention demonstrates a second use of an active ingredient (diethylcarbamazine- DEC) already known and used in treatment for filariasis (a medicine of choice) , which upon evaluation has been shown to also exhibit potent anti-inflammatory and anti-fibrotic action, especially when incorporated to a pharmaceutical and/or veterinary composition.

[050] The anti-inflammatory activity of DEC, or its pharmaceutically acceptable salts, were widely described in the literature, but the incorporation of this active principle into pharmaceutical or veterinary compositions were not reported, and after several experimental trials, we have been able to prove that the use of pharmaceutical preparations containing DEC, and/or its pharmaceutically acceptable salts, have an anti-inflammatory and anti-fibrotic activity superior to DEC administered alone, requiring a lower dose, in order to obtain superior results, besides prolonging the release time of DEC in the body. [051] In addition, many of the currently available anti ^¬ inflammatory drugs have severe adverse reactions, such as allergies, gastric toxicity, this invention may become an alternative for patients who are intolerant to conventional non-steroidal anti- inflammatory drugs (NSAIDs) .

[052] For example, the pharmaceutical and/or veterinary compositons of DEC, or its pharmaceutically acceptable salts, according to the present invention may be combined with other anti ^¬ inflammatory or immunomodulatory, anti-fibrotic, corticoid, analgesic, antitumor, antimycobacterial , fungicidal or antibiotic substances or agents, commonly used in combination or for treatment or prophylaxis of acute or chronic inflammatory diseases, and fibrotic processes. Most promisingly, the pharmaceutical composition of DEC, or its pharmaceutically acceptable salts, according to the present invention may also be combined with biological therapies, for example of antibodies and other recombinant proteins.

[053] The objective of the present invention is to provide alternatives for the treatment and/or prevention of acute and/or chronic inflammatory pathologies, which may be associated with fibrotic processes for human and/or animal use. In this sense, the present invention proposes the use of DEC, or its pharmaceutically acceptable salts, incorporated to pharmaceutical and/or veterinary compositions, in the manufacture of a medicament for the treatment or prevention of acute and/or chronic inflammatory pathologies, which may be associated with fibrotic processes.

[054] According to one incorporation of the present invention, it is described the use of DEC (Figure 1), or its pharmaceutically acceptable salts, incorporated to pharmaceutical and veterinary compositions, in the production of a medicament for the treatment or prevention of inflammatory pathologies, which may be associated with fibrotic processes. More specifically, the present invention describes the use of DEC, or its pharmaceutically acceptable salts, incorporated to pharmaceutical and veterinary compositions, in the production of a medicament for the treatment or prevention of acute and/or chronic inflammatory pathologies, associated or not to fibrotic processes.

[055] According to one incorporation of the present invention, it is described the use of a pharmaceutical and/or veterinary composition comprising: (a) DEC or its pharmaceutically acceptable salts and (b) pharmaceutically acceptable excipients for the production of a medication for treating disturbances or conditions in which inflammatory pathologies, which may be associated with fibrotic processes, predominates including the acute and chronic inflammations and fibrosis.

[056] According to another incorporation of the present invention, a method is provided for the treatment or prevention of acute and chronic inflammatory pathologies, which may be associated with fibrotic processes, comprising the administration of an effective amount of DEC and/or its pharmaceutically acceptable salts, incorporated to a pharmaceutical and/or veterinary composition, to human beings or animals.

[057] According to another incorporation of the present invention, a method is provided to treat and/or prevent acute and chronic inflammatory pathologies, associated or not with fibrotic processes, which comprises the administration of a pharmaceutical and/or veterinary composition comprising an effective amount of DEC and/or its pharmaceutically acceptable salts and excipients to human beings or animals.

[058] According to another incorporation of the present invention, a method is provided to treat and/or prevent acute and chronic inflammatory pathologies, associated or not with fibrotic processes, which comprises the administration and/or a combination of a pharmaceutical and/or veterinary composition comprising an effective amount of DEC and/or its pharmaceutically acceptable salts and excipients, with one or more other pharmacologically active substances, of one of the following therapeutic classes, not exclusively or limited to anti-inflammatory, anti-fibrotic, corticoid, analgesic, antitumor, antimycobacterial , fungicidal, antibiotic, immunomodulatory substances or agents, and biological therapies, for example of antibodies and other recombinant proteins, commonly used in combination or for treatment or prophylaxis of acute or chronic inflammatory diseases, and fibrotic processes to human beings or animals.

[059] The present invention also describes a pharmaceutical and/or veterinary composition comprising DEC and/or its pharmaceutically acceptable salts and excipients.

[060] The term "pharmaceutical and/or veterinary composition" as used in the present invention, refers to all pharmaceutical forms, and/or formulations containing one effective amount DEC and/or its pharmaceutically acceptable salts, associated with one or more pharmaceutically acceptable excipients, adjuvants, solvents and/or vehicles, without being limited to: solid form, for example, without being limited to: tablets (tablets uncoated or coated), tablets or films/wafers, which disintegrate rapidly into the oral cavity, sugar coated tablets, powders, granules, capsules (for example hard or soft gelatine capsules), films/lyophilisates, lozenges, pellets, capsules, pills, films, cachets, sugar-coated; or semi-solid form, for example, as ointments, creams, gels, suppositories, vaginal capsules, ova, emulsions, milk, pastes, foams, and/or therapeutic systems transdermal (eg adhesives); or in liquid form, for example as solutions, tinctures, sprays, aerosols, suspensions or emulsions. The pharmaceutical compositions of this invention may further be in the form of novel pharmaceutical forms including: liposomes, cyclodextrin complexes, nanospheres, nanocapsules , nanoparticles , microcapsules, microspheres, nanoemulsions and microemulsions . [061] The pharmaceutical and/or veterinary composition of the present invention may be administered by the following routes: oral, sublingual, nasal, rectal, intragingival , endovenous, intramuscular, intraarticular, subcutaneous, cutaneous (for example, patch or transdermal patch) , inhalatory, transdermal, topical or spinal (subarachnoid and peridural), parenteral, con unctival, optic, or as an implant or stent, not being limited to these. In particular, the oral route is preferred.

[062] The term "animals", as used in the present invention, refers to domesticated animals, wild animals kept in captivity or not, and laboratory animals.

[063] The term "effective amount", as used in the present invention, refers to an amount of diethylcarbamazine (DEC) , or its pharmaceutically acceptable salts, that provides the desired anti- inflammatory and/or anti-fibrotic activities when administered according to a dose appropriate for each administration route.

[064] The term "pharmaceutically acceptable excipients", as used in the present invention, refers to ingredients compatible, and that are not harmful to human beings or animals.

[065] The term " pharmaceutically acceptable salts", as used in the present invention, refers to salts which are selected from the group, but not limited to: alkali metal salts, (eg. sodium and potassium salts), alkaline earth metal salts (for example calcium and magnesium salts), chloride, bromide, hydrochloride, acetates, maleate, mesylate, nitrate, tartrate, gluconate, citrates, carbonates, carbamates, phosphates, diphosphates, phosphonates , glycosides, sulfates, sulfonates, and ammonium salts derived from amine or organic amines having from 1 to 16 carbon atoms.

[066] The term "to treat", as used in the present invention, refers to revert, alleviate, inhibit, prevent, or diminish the progress of inflammation and/or fibrosis in human beings or animals. The term "treatment", as used in the present invention, refers to the act of treating as defined above as well as to prevention. The terms "prevention" or "prophylaxis", were used in the present invention, refers to the act of preventing, which is related to the term "preemptive inflammation and/or fibrosis". This term refers to the preventive treatment of inflammation and/or fibrosis, before its occurrence. Such procedure has been often used post-operatively, when the responsible doctor knows that inflammation and/or fibrosis will arise after of surgical intervention or medical procedure.lt is also used in the treatment of cases of burns and chronic conditions, such as oncologic diseases, in which the doctor prescribes an anti-inflammatory regimen to be administered at fixed times, aiming at maintaining the integrity and functions of organs and tissues.

[067] The term "for the treatment and/or prevention of", as used in the present invention, refers to the treatment and/or prevention of one inflammatory and/or fibrotic diseases/conditions, selected from the following group, but not limited to: chronic obstructive disease, asthma, pulmonary hypertension, acute pulmonary inflammation such as respiratory distress syndrome (ARDS), pleuritis, hepatitis, cirrhosis, asthma, arthritis, arthrosis, systemic sclerosis, sclerodermatous graft-versus-host disease in bone marrow transplant recipients, Crohn's disease, cystic fibrosis, pulmonary fibrosis, intestinal fibrosis, liver fibrosis, kidney fibrosis, endomyocardial fibrosis, retroperitoneal fibrosis, and keloid.

[068] The characteristics of the present invention will become evident from the following detailed description.

[069] The present invention is directed to the use of DEC (Figure 1), or its pharmaceutically acceptable salts, incorporated to pharmaceutical and veterinary compositions, in the manufacture of a medicament for the treatment or prevention of inflammation and/or fibrosis pathologies/diseases. More specifically, the present invention describes the use of DEC, or its pharmaceutically acceptable salts, incorporated to pharmaceutical and veterinary compositions, in the prodution of a medicament for the treatment or prevention of acute and/or chronic inflammatory pathologies, which may be associated with fibrotic processes in human beings and animals

[070] According to another incorporation of the present invention, a method is provided for the treatment or prevention of acute and chronic inflammatory pathologies, which may be associated with fibrotic processes, comprising the administration of an effective amount of diethylcarbamazine or its pharmaceutically acceptable salts, incorporated to a pharmaceutical and/or veterinary composition, to human beings or animals.

[071] Another incorporation of the present invention refers to a pharmaceutical and/or veterinary composition containing an effective amount of diethylcarbamazine or its pharmaceutically acceptable salts, and pharmaceutically acceptable excipients for the use as described above. A pharmaceutical composition according to the present invention comprising 0.1% to 99% p/p of diethylcarbamazine or its pharmaceutically acceptable salts

[072] According to one incorporation of the present invention, it is described the use of a pharmaceutical and/or veterinary composition comprising: (a) diethylcarbamazine or its pharmaceutically acceptable salts and (b) pharmaceutically acceptable excipients for the production of a medication to treat disturbances or conditions in which inflammatory pathologies, which may be associated with fibrotic processes, predominates including the acute and chronic inflammations and fibrosis.

[073] According to the present invention, the administration of the pharmaceutical and/or veterinary composition of DEC and/or its pharmaceutically acceptable salts, may be performed by the following administration routes: oral, sublingual, nasal, rectal, intragingival , endovenous, intramuscular, intraarticular, subcutaneous, cutaneous (for example, patch or transdermal patch), inhalatory, transdermal, topical or spinal (subarachnoid and peridural), parenteral, con unctival, optic, or as an implant or stent, not being limited to these. In particular, the oral route is preferred .

[074] According to another incorporation of the present invention, the pharmaceutically acceptable excipients are selected according to the final presentation of the composition of the present invention, which may be in a non-exclusive manner, or limited to: in the form of capsules or tablets for oral administration, solutions for nasal administration, injectable solutions for intramuscular, endovenous, cutaneous or subcutaneous administration, and lotion, cream or ointment for topical administration.

[075] Methods for the preparation of several pharmaceutical and veterinary compositions are well known, or will be recognized in the light of the present invention, by the art specialist in pharmaceutical technology, or may be obtained for example from: United States Pharmacopoeia, European Pharmacopoeia, British Pharmacopoeia, Brazilian Pharmacopoeia, Remington's Pharmaceutical Sciences, 18th ed. (1990) .

[076] According to another incorporation of the present invention, a method is provided to treat and/or prevent acute and chronic inflammatory pathologies, which may be associated with fibrotic processes, which comprises the administration and/or a combination of a pharmaceutical and/or veterinary composition comprising an effective amount of diethylcarbamazine or its pharmaceutically acceptable salts, and pharmaceutically acceptable excipients, with one or more other pharmacologically active substances, commonly used in combination or for treatment or prophylaxis of acute or chronic inflammatory diseases, and fibrotic processes to human beings or animals, belonging to one of the following therapeutic classes, not exclusively or limited to anti ^¬ inflammatory, or immunomodulatory, anti-fibrotic, corticoid, analgesic, antitumor, antimycobacterial , fungicidal, antibiotic substances or agents, and biological therapies, for example of antibodies and other recombinant proteins.

[077] According to the outcomes of the following tests, DEC and/or its pharmaceutically acceptable salts, incorporated into a pharmaceutical composition (generally called NANO-DEC) , exhibits greater anti-inflammatory and anti-fibrotic response than DEC administered alone.

[078] In in vivo model of acute inflammation demonstrates that DEC, and/or its pharmaceutically acceptable salts, incorporated into a pharmaceutical composition (NANO-DEC) at 12.5 mg/kg (NANO-DEC 12.5) has a better inflammatory response than does DEC administered alone at 50 mg/kg (DEC 50), taking into account the following parameters: (a) histopathological evaluation. (b) evaluation of the hepatic glutathione reductase activity; and (c) evaluation of the inflammatory proteins ( cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS)) expressed by Western Blot. [Rodrigues et al . , 2015]

[079] In the histopathological evaluation, the group treated with DEC 50 was not able to decrease the presence of inflammatory infiltrate, nor prevent the death of several hepatocytes, and promote only a small decrease in the necrosis areas. The group treated with NANO-DEC 12.5 showed a significant decrease in the necrosis and inflammatory infiltrate areas, and the liver tissue presentd similar morphology found in the control group. In the evaluation of hepatic glutathione reductase activity the group treated with DEC 50 and the group treated with NANO-DEC 12.5 were able of stimulating the enzymatic activity, however, the group treated with NANO-DEC 12.5 promoted greater glutathione reductase activity than DEC 50. In the evaluation of the expression of COX-2 and iNOS enzymes, the group treated with DEC 50 was only able to reduce significantly the COX-2 expression, whereas the group treated with NANO-DEC 12.5 significantly reduced the expression of the inflammatory markers COX-2 and iNOS . [Rodrigues et al . , 2015]

[080] In in vivo model of chronic inflammation, associated with fibrosis processes demonstrates that DEC, and/or its pharmaceutically acceptable salts, incorporated into a pharmaceutical composition the groups treated with (NANO-DEC) at 12.5 mg/kg (NANO-DEC 12.5) has a better anti-inflammatory response than does the group treated with DEC administered alone at 50 mg/kg (DEC 50), taking into account the following parameters: (a) histopathological evaluation; (b) evaluation of the collagen content by Sirius red staining; and (c) evaluation of the inflammatory proteins (transforming growth factor beta (TGF-β) and interleukin 1 beta (IL-Ιβ)) expressed by Western Blot.

[081] In the histopathological evaluation, the group treated with DEC 50 presented a reduction in the inflammatory infiltrates and vacuolization, however many areas of necrosis were still present. The group treated with NANO-DEC 12.5 there was a completely reduction in all histological alterations observed in the CCI4 group, with absence of vacoalization and of necrosis areas, restoring the standard hepatic morphology. In the evaluation of the expression of TGF-β and IL-Ιβ, the DEC 50 group was only able to reduce significantly the TGF-β expression when compared to control group, while the treatment with NANO-DEC 12.5 significantly reduced the expression of both TGF-β and IL-Ιβ (Figure 10) .

[082] In the evaluation of the collagen content by Sirius red staining, the group treated with DEC 50 exhibited significant reduction of red staining, presenting staining in the perilobular region and in the sinusoids. However, the group treated with NANO- DEC 12.5 exhibited a greater reduction in the labeling of collagen fibers, with marked improvement in the morphological aspect.

[083] The following examples are simply illustrative, and here placed only for the purpose of clarifying, as well as to provide a better understanding of the developments in the present invention, although they should not be used with the intention of limiting the described objects.

[084] Preparation and characterization of pharmaceutical composition containing diethylcarbamazine or its pharmaceutically acceptable salts:

[085] The following examples describes one of the possible pharmaceutical composition, where DEC, or its pharmaceutically acceptable salts, can be incorporated in order to increase the anti ^¬ inflammatory and/or anti-fibrotic activities of DEC.

[086] The preparation of polymeric nanoparticles containing DEC citrate, determination of particle size and zeta potential, morphological characterization of the formulation and release kinetics, and the determination of diethylcarbamazine amount in the composition, are described.

[087] Example 1 - Preparation and characterization of polymeric nanoparticles containing DEC-citrate

[088] DEC nanosystems compositions may be prepared through the technique of interfacial polymerization in an emulsified system hydrophilic/lipophilic/hydrophilic (w/o/w) type, as follows, in order to obtaining polymeric nanoparticles, containing DEC, pharmaceutically acceptable excipients and carriers:

[089] Polymeric nanoparticles containing DEC citrate: nanoparticles formed from the formation of a multiple emulsion w/o/w (water/oil/water) followed by the solvent evaporation, where the aqueous phase 1 comprises DEC citrate and water; and the organic phase comprises a hydrophobic polymer dissolved in an organic solvent and a low EHL surfactante, and the second aqueous phase comprises a stabilizer and water.

[090] The nanoparticles preparation process consists of: the DEC citrate solubilization in water; the solubilization of the hydrophobic polymer and the low EHL surfactant in organic solvent, for example dichloromethane (so-called organic phase) . The mixture of these two solutions provide the first water/oil emulsion. Then this primary emulsion is added to a water phase and stabilizers under intense agitation ( sonification) to form the DEC-containing nanoparticles . Then the organic solvent (in this example, the dichloromethane) is removed under reduced pressure, for example with the aid of a rotary evaporator.

[091] Aqueous suspensions containing the DEC nanoparticles of the present invention may be obtained by varying the concentrations of the active ingredient, excipients and pharmaceutical carriers in the ranges of: 0.1 to 99% of DEC and/or its pharmaceutically acceptable salts; 0.01 to 30% hydrophobic polymer; 0.01 to 30% surfactant; 0.01 to 30% stabilizer; and 0.01 to 90% water.

[092] Table 1, describes an example of a specific DEC composition obtained. However, it should be understood that such examples are presented only for illustrative purpose, and only to prove the success of the present invention. Any modifications in the light of the formulation herein, with percent active ingredient (DEC) percentages, percent excipients, solvents, or pharmaceutical carriers used for the preparation of nanoparticles containing DEC, are within the scope of this invention and its claims.

[093] Table 1: Materials (DEC, solvents, excipients, and vehicles) used to prepare the suspension of DEC nanoparticles.

[094] Table 1. Characterization of suspension formulation of DEC nanoparticles.

Component Amount

Aqueous phase 1

Water lmL

Diethylcarbamazine 25mg

Organic phase

Dichlorometane lOmL

Poly ε-caprolactone 50mg Pluronic F68 30mg

Aqueous phase 2

Polivinylalcool 200mg

Water 40mL

[095] In order to confirm the nanometric and unimodal size of the nanometer DEC suspension, the average size and the polydispersion index (PDI) of the DEC-containing nanoparticles was determined by laser diffraction using NanoZS 90 (Malvern) equipment. For analysis the samples were diluted in ultra pure water in the ratio of 1:20. The zeta potential was measured using the same equipment, but the suspensions were diluted in 1 mMNaCl solution.

[096] The DEC-containing nanoparticles had a mean size of 315 ± 5 nm with a PDI of 0.2 and unimodal distribution; the zeta potential was -5 ± 1.2 mV. These same values were obtained after the lyofilization of the NANO-DEC and after a period of strage of 180 days .

[097] For the study of the DEC release profile in the DEC- containing nanoparticles the dialysis method was used. 10 mL of the formulation was added in dialysis bags and the amount of DEC released in the medium was quantified by spectrophotometry, at predetermined times .

[098] The release profile demonstrated that the formulation took 6 hours to release 64% of the drug (DEC) incorporated into the composition (Figure 2); and a total time of 24 hours to release 100% of the drug (DEC) in the medium.

[099] The encapsulation efficiency ranged from 72 to 90%.

[100] The morphological characterization of the formulation was carried out using a Transmission Electron Microscope (TEM) . The formulation was deposited on copper supports and negatively stained negatively. After drying, the formulation was microphotographed . [101] The TEM analysis demonstrated that the nanoparticles presentd in spherical or oblong and individualized forms, according to Figure 3.

[102] In vivo assays for acute in lammation:

[103] In order to prove, the greater acute anti-inflammatory activity of diethylcarbamazine (DEC) , and/or its pharmaceutically acceptable salts, incorporated into a pharmaceutical composition (in this invention, generally called NANO-DEC) , when compared to DEC administered alone, ain vivo model of acute hepatic inflammation induced by carbon tetrachloride (CCI4) was performed.

[104] To perform the in vivo model of acute hepatic inflammation induced by carbon tetrachloride (CCI4) a total of 28 ( twenth-eight ) male C57BL/6 mice were used. DEC or NANO-DEC was given intraperitoneally for 6 days (1 in ection/day) . For the acute liver inflammation model the animals were treated with 0.1 μΐ/g body weight of CCI4 ( Sigma-Aldrich, St Louis, MO, USA) administered in a single dose on the sixth day of treatment with DEC or NANO-DEC. CCI4 was diluted in olive oil and administered intraperitoneally. The animals were divided into four experimental groups (n = 7 animals/group) : Control (received only feed and water) , CCI4 (received only CCI4) , CCI4 + DEC 50 (received DEC citrate at 50 mg/kg and CC1 ₄ ) , CC1 ₄ + NANO-DEC 12.5 (received DEC citrate DEC-containing nanoparticles at a concentration of 12.5 mg/kg and CCI4) .

[105] The following parameters related to the acute inflammation model were evaluated: histopathological evaluation, evaluation of the hepatic glutathione reductase activity, and the evaluation of the inflammatory proteins ( cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS)) expressed by Western Blot.

[106] Example 2 - Histopathological Evaluation

[107] The hepatic fragments were fixed in 10% formalin solution for 24 hours. The tissue was then dehydrated in ethanol and xylol solutions and embedded in paraffin. Cuts of 4 μπι thickness were performed and stained by the Hematoxylin - Eosin (H.E.) technique. In figure 4, we observed the histopathological changes of the analyzed groups.

[108] It was observed that the control group had standard liver morphology. The CCI4 group presentd severe damage in the hepatic stroma, with extensive areas of necrosis and of massive inflammatory infiltrate. In the CCI4 + DEC 50 group, a small decrease in the necrosis areas was observed, but still with the presence of inflammatory infiltrate and high number of hepatocyte death. The CCI4 + NANO-DEC 12.5 group showed a significant decrease in the areas of necrosis and inflammatory infiltrate, being close to the morphology found in the control group (Figure 4) .

[109] Example 3 - Evaluation of the Hepatic Glutathione Reductase Activity

[110] Hepatic liver fragments were obtained and weighed. Each fragment was homogenized using a manufacturer's buffer (Abnova Corporation, Taipei, Taiwa, catalog: KA0881) and centrifuged at 10, 000 x for 15 minutes at 39,2 °F. The supernatant was used to estimate the activity of hepatic glutathione reductase through the assay kit following the manufacturer's instructions.

[Ill] Glutathione reductase catalyzes the reduction of oxidized glutathione (GSSG) to glutathione (GSH) , an essential step in the glutathione cycle. This last one, in turn, promotes the defense of cells against oxidative stress.

[112] A significant decrease in glutathione activity in the CCI4 group was observed in relation to the control group. However, DEC seems to be a stimulant of enzyme activity, since both the CCI4 + DEC 50 group and the CC1 ₄ + NANO-DEC 12.5 group were able to significantly increase enzyme activity. However, the NANO-DEC 12.5 group obtained greater glutathione reductase activity than the DEC 50 group (Figure 5) . [113] Example 4 - Evaluation of the In lammatory Proteins expressed by Western Blot

[114] Hepatic fragments were dissected and homogenized in extraction cocktail. Proteins (40mg) were separated on SDS gels by gel electrophoresis and then electrophoretically transferred to a nitrocellulose membrane (BioRad, CA, USA) . After overnight blocking at 39.2 °F, the membranes were incubated at room temperature for 4 h with the primary polyclonal antibodies. Anti-rabbit IgG secondary antibody conjugated with horseradish peroxidase (HRP) (1:1000) was used. The blots were revealed on X-ray film (Fuji Medical, Kodak), and quantified using the ImageJl.38 program®. For each protein investigated, the results were confirmed in duplicate.

[115] The expression of two enzymes involved in the inflammatory process was evaluated. Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) . It was observed that the CCI4 group showed a significant increase of both enzymes in relation to the control group (Figure 6) .

[116] The CCI4 + DEC 50 group had reduced expression of COX-2 and iNOS, however, only COX-2 enzyme expression was significant when compared with the group treated with CCI4. In contrast, the group treated with DEC-containing nanoparticle (NANO-DEC 12.5) significantly reduced the expression of COX-2 and iNOS in relation to the CCI4 group (Figure 6) .

[117] In vivo assays for chronic inflammation, and fibrosis.

[118] In order to prove, the greater chronic anti-inflammatory and anti-fibrotic activities of diethylcarbamazine (DEC) , and/or its pharmaceutically acceptable salts, incorporated into a pharmaceutical composition (in this invention, generally called NANO-DEC) , when compared to DEC administered alone an in vivo model of chronic hepatic inflammation induced by carbon tetrachloride (CCI4) was performed. [119] To perform the in vivo model of chronic hepatic inflammation induced by carbon tetrachloride (CCI4) , the experimental groups was composed by a total of 42 (forty-two) male C57BL/6 mice (age: 5 weeks; weight: 15-17 g) were housed in metal cages and fed with a standard diet and water ad libitum. Chronic inflammation was induced by intraperitoneal administration of CCI4 0.5 of body weight ( Sigma-Aldrich, St. Louis, MO, USA) dissolved in olive oil (final volume, 100 \i per mouse) . Two CCI4 injections were administered per week for 8 weeks. DEC (25 mg/kg and 50 mg/kg) and NANO-DEC (12.5 mg/kg and 5 mg/kg) were administered through intraperitoneal injection for 6 days (1 injection/day) on the last six days of the CCI4 administration. The control group received distilled water via the same route. The C57BL/6 mice were separated into six groups (n= 7/group) : (1) the control group (Cont) , which received just distilled water; (2) the CCI4 group (CCI4) ; (3) the DEC 25 mg/kg plus CC1 ₄ group (CC1 ₄ + DEC 25); (4) the DEC 50 mg/kg plus CCI4 group (CC1 ₄ + DEC 50) ; (5) the NANO-DEC 05 mg/kg plus CC1 ₄ group (CCI4 + NANO-DEC 05) and (6) the NANO-DEC 12.5 mg/kg plus CC1 ₄ group (CCI4 + NANO-DEC 12.5) . 24 h after the last intraperitoneal administration of CCI4, the animals were anesthetized (xylazine 10 mg/kg and ketamine 100 mg/kg) and sacrificed by cervical dislocation.

[120] The following parameters related to the chronic hepatic inflammation model were evaluated: histopathological evaluation; evaluation of the collagen content by Sirius red staining; and the evaluation of the inflammatory proteins (transforming growth factor beta TGF-β and interleukin 1 beta (IL-Ιβ) expressed by Western Blot.

[121] Example 5 - Hematoxylin-eosin staining

[122] Liver fragments were fixed in 10% formalin for 24 h, processed, and embedded in paraffin. Sections of 4-5 μπι were cut and mounted on glass slides. Slices were stained with hematoxylin-eosin and assessed by inverted microscopy (Observer Zl; Zeiss Microimaging, Jena, Germany), a camera, and 4.7.4 image analysis software (AxionCamMRm; Zeiss, Jena, Germany) at a magnification of ^χ 400.

[123] Histological analysis of the control group displayed the standard hepatic morphology. In the group of animals exposed to CCI4, various histological characteristics of persistent inflammation were observed. This group showed striking cytoplasmic degeneration, several fibrosis areas, vacuolization and massive inflammatory infiltrates. The same finds were observed in CCI4 + DEC 25 and CCI4 + NANO-DEC 5 groups. Animals exposed to CCI4 and treated with DEC 50 mg/kg presetend a reduction in the inflammatory infiltrates and vacuolization, however many areas of necrosis were still present. In the histology of the CCI4 + NANO-DEC 12.5 group, there was a reduction in all of the histological alterations observed in the CCI4 group. There was a noticeable decrease in vacoalization and in the areas of necrosis. Furthermore, the inflammatory infiltrates were reduced, and the hepatocytes returned to the standart morphology ( Figure 7 ) .

[124] Example 6 - Assessment of collagen content by Sirius red staining .

[125] Liver fragments were fixed in 10% formalin for 24 h, processed, and embedded in paraffin. Sections of 4-5 μπι were cut and mounted on glass slides. Slices were stained with picrosirius red solution of 1%, rehydrated in water, and stained with Fast Green 0.1% solution in ethanol. They were then assessed by inverted microscopy (Observer Zl; Zeiss Microimaging), a camera and 4.7.4 image analysis software (AxionCamMRm; Zeiss) at a magnification of x400.

[126] In the assessment of collagen content by Sirius red staining, the control group exhibited little staining of collagen fibers, which was restricted to the periphery of the centrilobular veins. CCI4 group exhibited markedly red staining, indicating a high presence of collagen fibers throughout the hepatic stroma. Similar results were observed in CCI4 + DEC 25 and CC1 ₄ + NANO-DEC 5 groups. The CC1 ₄ + DEC 50 exhibited significant reduction of red staining, presenting staining in the perilobular region and in the sinusoids. The CCI4 + NANO-DEC 12.5 group exhibited a considerable reduction in the labeling of collagen fibers, with marked improvement in the morphological aspect (Figures 8 and 9) .

[127] Example 7 - Evaluation of the in lammatory proteins (TGF- β and IL-Ιβ) expressed by Western Blot

[128] Livers were quickly dissected and then homogenized in a

Wheaton Overhead Stirrer (model 903475; Wheaton Science Products, Millville, NJ, USA) using an extraction cocktail of 10 mmol/L ethylenediaminetetra acetic acid, 2-mmol/L phenylmethylsulfonylfluoride, 100 mmol/L

sodium fluoride, 10-mmol/L sodium pyrophosphate, 10-mmol/L sodium orthovanadate, 10-mg aprotinin, and 100-mmol/L Tris (hydroxymethyl ) aminomethane (pH 7.4) . Western blot assays were then performed according to the previously described methodology [1] . The antibodies used were anti-IL-Ιβ (1:1000 dilution; ab9722; Abeam) and anti-TGF-βΙ (1:1000 dilution; sc-109; Santa Cruz) .

[129] In the western blot analysis, the CCI4 group exhibited an elevated expression of TGF-β and IL-Ιβ when compared with the control group. The CC1 ₄ + DEC 25 and CC1 ₄ + NANO-DEC 5 groups also presentd high expression when compared with the control group. The treatment with DEC 50 and NANO-DEC 12.5 significantly reduced the expression of the TGF-β cytokine. However, only NANO-DEC 12.5 treatment was able to reduce the expression of IL-Ιβ (Figure 10) .

[130] DESCRIPTION OF THE FIGURES

[131] Descriptions of the figures that accompany this detailed description are present below, for better understanding and illustration of the present invention.

[132] Figure 1. Chemical Structure of Diethylcarbamazine (DEC) . [133] Figure 2. Graph of the release profile of DEC-containing nanoparticle in the formulation. A- The X-axis refers to the Time in minutes. B- Axis Y refers to the amount of drug released in the medium (mg) .

[134] Figure 3. Transmission Electron Microscopy (TEM) of an individualized DEC-containing nanoparticle in the formulation.

[135] Figure 4. Histopathological analysis of hepatic fragments stained with H/E.

[136] Figure 5. Activity of hepatic glutathione reductase. Results were expressed as a mean ± SEM of four animals per group. * P <0.05 vs. CCI4; # P <0.05 vs control. 1U = amount of enzyme generating 0.5 μπιοΐ of NADP per minute at 77 °F.

[137] Figure 6. Immunoblot of inflammatory markers. A - COX-2; B - iNOS . The densitometric analysis was performed using Image J 1.38 software. Results were expressed as a mean ± SEM of five animals per group. * P <0.05 vs. CC1 ₄ ; #P <0.05 vs. Control.

[138] Figure 7. Histological analysis of liver fragments in haemotoxylin-eosin staining. Magnification: 400x. Liver thin section = 4-5μπι.

[139] Figure 8. Assessment of the content of collagen fibres by

Sirius red staining.

[140] Figure 9. Quantification (five arbitrarily selected areas per animal) was performed using GIMP 2 image analysis software (n=4 animals per group) . The results are expressed as mean ± S.D. *P<0.05 vs CC14; # P<0.05 vs control.

[141] Figure 10. Immunoblot of inflammatory markers. The densitometric analysis was performed with IMAGEJ 1.38 software (n=5 animals per group) . The results are expressed as mean ± S.D. *P<0.05 vs CC14; # P<0.05 vs control.