WO2013160492A1 - Combination therapy for the treatment of metabolic diseases - Google Patents

Abstract

The invention relates to the use of beta-3 adrenergic receptor agonists in a combination therapy for the treatment of metabolic diseases, such as hyperglycemia, insulin resistance, type 2 diabetes, and dyslipidemia. This therapy combines the administration of a beta-3 adrenergic receptor agonist with a GLP-1 receptor agonist or a PPARα agonist, both compounds being capable of reducing appetite, reducing weight gain and increasing energy expenditure by means of fat mobilisation. The invention demonstrates that the combined administration of said compounds has a synergistic effect on the reduction of body fat.

Description

 COMBINED THERAPY FOR THE TREATMENT OF DISEASES

 METABOLICS

TECHNICAL FIELD OF THE INVENTION

 The present invention relates to methods for the treatment and / or prevention of obesity and dyslipidemia. More specifically, the methods and uses of the invention relate to the administration of a stable derivative of an adrenergic beta-3 analog in combination with the administration of a PPARa ligand (peroxisome proliferating activated receptor type alpha) or a receptor agonist. GLP-1 (glucagon like peptide 1).

BACKGROUND OF THE INVENTION

The incidence of obesity in European countries is a major problem for health systems due to its enormous implications for health, mortality and quality of life. Obesity is the biggest risk factor for cardiovascular disease and type 2 diabetes, and is also independently associated with an increase in mortality and morbidity. Obese individuals die prematurely due to coronary and cerebrovascular diseases, as well as due to certain types of cancer. They also suffer from substantial morbidity and reduced quality of life due to musculoskeletal and gastrointestinal pathologies, depression and social isolation and poor mobility. Currently, existing pharmacological treatments are no more effective than diet. In fact, the pharmaceutical industry has failed to find new drugs capable of reducing weight steadily with minimal side effects.

 Some of the pharmacological methods currently available for the treatment of obesity are:

Orlistat (Xenical) is a weight loss medication that inhibits gastrointestinal lipase by blocking digestion and fat absorption. It produces a moderate weight loss when combined with diet and physical exercise for one or two years, but it has gastrointestinal side effects, including steatorrhea, fecal incontinence and frequent or urgent bowel movements.

 - Phentermine (phenyl-tertiary-butylamine) is a psychostimulant drug of the phenylethylamine class that acts as an appetite suppressant. It is normally tolerated properly, but is only indicated for short-term treatments, because it can cause dependence, so it does not usually result in long-term weight loss.

 Sibutramine (Meridia) is a centrally acting serotonin-norepinephrine reuptake inhibitor, and makes the patient feel satiated more quickly. It produces great weight loss when used in conjunction with a healthy diet and exercise program, but its use has been associated with an increase in cardiovascular events and strokes, so it has been withdrawn from the market in different countries and regions , including Australia, Canada, China, the European Union, Hong Kong, India, Mexico, Thailand, the United Kingdom and the United States.

 - Finally, Rimonabant, an endocannabinoid receptor antagonist, has severe side effects related to alterations in emotional behavior such as depression, which is why it has been withdrawn from the market.

The increase in energy expenditure through the conversion of fats (lipids) into heat, a process called thermogenesis, is a cleaner and safer process to lose weight than the pharmacological reduction of appetite or chemical breakdown of fat, which generates potentially dangerous by-products in significant concentrations. Pharmacological induction of thermogenesis can be achieved by activating a class of adipose tissue called brown adipose tissue. This tissue is made up of cells that have a large amount of a special type of mitochondria and small lipid droplets; These mitochondria are able to consume energy to produce heat thanks to a mitochondrion-specific decoupling protein (Riquier et al, Reprod Nutr Dev, 1985, 25: 175-181), which decouples oxidative phosphorylation and allows brown adipose tissue to become a "heat generating gland." As would be expected in rats, this protein is produced in large quantities after exposure to cold (Bouillaud et al, J Biol Chem, 1984, 259: 11583-11586). Under certain conditions, white adipose tissue is capable of become brown adipose tissue, which is accompanied by the appearance of the typical decoupling protein in the inner mitochondrial membrane. In patients with pheochromocytoma (a tumor that secretes epinephrine and other catecholamines), the perirenal fat may change from white to brown (Lean et al, In J Obesity 1986, 10: 219-227; Melicow, Arch Pathol 1957, 63: 367- 372). The same effects are observed in rats (Ricquier et al, Reprod Nutr Dev 1985, 25: 175-181; Ricquier et al, J Am J Physiol 1983, 254: C172-177).

 The activation of a particular class of epinephrine receptor, the beta-3 adrenergic receptor, has emerged as a target for pharmaceutical drug development that increases thermogenesis. Adrenergic beta-3 receptors are mostly expressed in adipocytes. Stimulation of beta-3 adrenergic receptors induces lipolysis in white adipocytes and non-thyring thermogenesis in brown adipocytes. Beta-3 adrenergic receptor agonists are able to prevent or reverse obesity in animal models. Despite these promising qualities, the pharmaceutical industry has not been successful in developing a beta-3 adrenergic receptor agonist for use in the treatment of human obesity and type 2 diabetes. The high doses of agonists needed to stimulating human beta-3 adrenergic receptors produce unwanted effects; It is also unclear whether long-term stimulation of beta-3 adrenergic receptors is safe.

 Therefore, there is a need in the art for additional therapies based on beta-3 adrenergic receptor agonists that do not have the disadvantages of therapies known in the state of the art.

SUMMARY OF THE INVENTION

 In a first aspect, the invention relates to a composition or a kit of parts comprising, together or separately, two elements: a first element that is an adrenergic beta-3 receptor agonist and a second element that can be an agonist of the GLP-1 receptor, a PPARa agonist or a combination of both.

In a second aspect, the invention relates to the use of the composition or kit of parts of the invention for the preparation of a medicament for the treatment of a metabolic disease. In a third aspect, the invention relates to a cosmetic method for the treatment of obesity and / or cellulite, which comprises administering to a patient a pharmacologically active amount of a composition or kit of parts with the characteristics indicated in the first aspect. of the invention.

BRIEF DESCRIPTION OF THE FIGURES

 Figure 1: (A) Chemical structure of the beta-3 agonist CL-316243. (B) Activity of the beta-3 agonist CL-316243 as an intake suppressant. Fasting food intake was tested on fasting animals 30 min, 60 min, lh, 2h, 4h, 6h, 8h, 12h and 24h after the i.p. of the compound at different doses (0.1 mg / kg, 0.5 mg / kg and 1 mg / kg). CL316243 is a powerful intake suppressant. The results are means ± SEM of at least three determinations per group. (*) p <0.05, (**) p <0.01, (***) p <0.001, in front of the vehicle, ANOVA. (C) Dose-response curve of the reduction of circulating triglycerides by CL316243.

Figure 2: Effects of subchronic treatment with CL316243, liraglutide and a combinatorial treatment of CL-316243 and liraglutide on (A) inhibition of intake, (B) amount of total lipid mass and (C) triglycerides. Subchronic treatment (6 days, ip of CL-316243 once a day and subcutaneous injection of liraglutide 2 times a day) reduces food intake (A), total lipid mass (B) and plasma triglyceride content ( C) when compared to normal rats treated with vehicle. A clear synergistic effect of combinatorial treatment can be observed. The results are means ± SEM of at least three determinations per group. (***) p <0.001, against vehicle, ANOVA.

Figure 3: Effects of subchronic treatment with CL-316243, oleoylethanolamide (OAS) and a combinatorial treatment of CL-316243 and OAS on (A) inhibition of intake, (B) amount of total lipid mass and (C) triglycerides. Subchronic treatment (6 days, a daily intraperitoneal dose) reduces food intake (A), total lipid mass (B) and plasma triglyceride content (C) when compared to normal vehicle-treated rats. A clear synergistic effect of combinatorial treatment can be observed. The results are means ± SEM of at least three determinations per group. (*) p <0.05, (**) p <0.01, (***) p <0.001, versus vehicle, ANOVA. Figure 4: Effect of treatment with CL-316243, oleoylethanolamide and a combinatorial treatment of CL-316243 and oleoylethanolamide on the expression of the UCP1 gene in white adipose tissue. Combined therapy significantly increases the expression of UCP1 RNA.

Figure 5: Effect of treatment with CL-316243, oleoylethanolamide and a combinatorial treatment of CL-316243 and oleoylethanolamide on the expression of the UCP1 gene in brown adipose tissue. Combined therapy significantly increases the expression of UCP1 both at the RNA level (A) and at the protein level by immunohistochemistry (B).

DETAILED DESCRIPTION OF THE INVENTION

 The authors of the present invention have observed that, surprisingly, the administration of an adrenergic beta-3 receptor agonist in combination with GLP-1 receptor agonists or PPARa agonists (both compounds capable of reducing appetite and weight gain and to increase energy expenditure through fat mobilization), they produce a significant reduction in food intake, body fat and circulating triglycerides, with a clear synergistic effect.

Compositions of the invention

Thus, in a first aspect, the present invention relates to a composition or kit of parts comprising, together or separately, an adrenergic beta-3 receptor agonist in combination with a GLP-1 agonist or a PPARa agonist for The treatment of a metabolic disease. Additionally, the invention relates to the use of said composition in the preparation of a drug for the treatment of a metabolic disease as well as a method for the treatment of said metabolic disease, which comprises the administration to a subject of a drug comprising said drug. composition.

By composition, as used in the present invention, it refers to a material composition comprising the aforementioned components, as well as any product resulting, directly or indirectly, from the combination of the different components in any amount thereof. The composition can be formulated as a single formulation or may be presented as separate formulations of each of the components, which may be combined for joint use as a combined preparation.

 By kit of parts, as used in the present invention, is meant a composition in which each of the components are formulated individually. Additionally, said components can be packaged individually. Thus, the components of the parts kit do not need to be present in a combined form, such as in a true composition, but can be formulated separately for their combined, separate or sequential application, in view of the physical separation of the components.

 By "beta-3 adrenergic receptor" is meant the protein defined by the sequence described in the UniProt database by the accession number P 13945. It is a type of beta adrenergic receptor, which mediates the activation of induced adenylate cyclase by catecholamines through the activation of G proteins. The beta-3 adrenergic receptor is mostly expressed in adipose tissue, where it is involved in the regulation of lipolysis and thermogenesis.

 The term "beta-3 adrenergic receptor agonist" refers to a compound that binds to the beta-3 adrenergic receptor and activates it. The activity of a compound as an adrenergic beta-3 receptor agonist can be assessed by the adenosine monophosphate accumulation assay (Takasu et al., J Pharmacol Exp Ther., 2007, 321: 642-647) or by the ability to stimulate the production of adenyl cyclase activity in adipocytes as described by Cooper et al. (J. Biol. Chem., 1979, 254: 8927-8931).

 In a preferred embodiment, the beta-3 adrenergic receptor agonist is a compound with the formula

or a pharmaceutically acceptable derivative salt, wherein

R4 is a hydroxy, alkoxy, amino, alkylamino or dialkylamino

R ₅ is hydrogen, fluorine, chlorine, bromine, iodine, -CN, CF ₃ , short chain alkyl, short chain alkoxy, cycloalkyl or aryl;

 5 is short chain alkyl, cycloalkyl or aryl;

R ₇ , R ₇ ', R ₈ and R ₈ ' can independently hydrogen, alkyl short chain or R ₇ and R ₈ can be together CH ₂ -CH ₂ ; hydrogen or

 m is an integer between 1 and 2;

 n is zero or an integer between 1 and 6,

 p is an integer between 1 and 5

The term "alkyl" refers to both linear and branched chain groups, with 1-12 carbon atoms in the normal chain, preferably 1-7 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, t- butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, various branched chain isomers derived, and the like.

 The term "short chain alkyl", as used herein, includes alkyl groups as mentioned above comprising from 1 to 6 carbon atoms in its chain.

 The term "alkoxy" refers to any of the aforementioned alkyl groups attached to an oxygen atom.

 The term "short chain alkoxy" refers to any of the aforementioned short chain alkyl groups attached to an oxygen atom.

 The term "aryl" refers to a monocyclic or bicyclic aromatic group comprising from 6 to 10 carbon atoms in the ring portion, such as phenyl, naphthyl, substituted phenyl or substituted naphthyl, where the substituent both in the phenyl and in the naphthyl may be 1, 2 or 3 short chain alkyl groups, halogens (for example chlorine, bromine, fluorine) or 1, 2 or 3 short chain alkoxy groups.

 The term "cycloalkyl" refers to saturated hydrocarbon cyclic groups containing one or more rings of 3 to 12 carbons in total in the ring, preferably 3 to 8 carbons in the ring, which includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, and adamantyl.

The term "alkylamino" refers to an amino group that has an alkyl group attached. The term "dialkylamino" refers to an amino group that has two alkyl groups attached, which may be the same or different.

 In another preferred embodiment, the beta-3 adrenergic receptor agonist is CL-316243. The term "CL-316243" ((R, R) -5- [2- [2- (3- chlorophenyl) -2-hydroxyethylamino] propyl] -l, 3-benzodioxol-2,2-dicarboxylic acid) refers to to a compound whose formula is

Compound CL-316243 is a highly selective beta-3 adrenergic receptor agonist capable of increasing lipolysis, fatty acid oxidation and insulin action in humans.

 In another embodiment, the beta-3 adrenergic receptor agonist is a member of the following list: BRL 37344, CPG 12177, CL-316243, SR 58661, pindolol, cyanopindolol, YM178, GW427353, TRK-380,

"GLP-1" (Glucagon-like peptide-1) means the gastrointestinal peptide derived from the transcription product of the proglucagon gene, defined by the sequence described in the UniProt database by the accession number P01275. GLP-1 is released by cells of the intestine and endocrine pancreas in response to the elevation of certain levels of certain nutrients in plasma, such as glucose. It is a hormone with a potent anti-hyperglycemic effect, inducing a glucose-dependent insulin secretion while suppressing glucagon secretion. Such glucose-dependent action is particularly attractive because, when fasting blood glucose values are normal, GLP-1 does not stimulate insulin secretion causing hypoglycemia. GLP-1 seems to restore the glucose sensitivity of pancreatic beta cells, through a mechanism that possibly implies an increase in the expression of GLUT2 and glucokinase. GLP-1 also inhibits apoptosis of pancreatic beta cells and stimulates the proliferation and differentiation of insulin-secreting beta cells. Additionally, GLP-1 inhibits secretion and gastric motility, which delays and prolongs carbohydrate absorption and contributes to a satiating effect.

 The term "GLP-1 receptor agonist," as used in the present invention, refers to any GLP-1 mimetic, including GLP-1 receptor agonists (exenatide), structurally modified peptides (liraglutide), analogs. conjugates (CJC-1134-PC and naliglutide or albugon).

 The activity of a compound as a GLP-1 receptor agonist can be assessed by an assay based on the ability of the agonist to induce insulin secretion by isolated pancreatic islets as described by Knudsen et al., Proc. Nat.Acad.Sci.USA, 2006, 104: 937-942.

 In a preferred embodiment, the GLP-1 agonist is liraglutide.

In another possible embodiment, the GLP-1 receptor agonist is a member of the following list: exenatide; lixisenatide; dulaglutide; albiglutide; taspoglutide; exendin-3; Leu ¹⁴ -exendina -4; Leu ¹⁴ , Phe ²⁵ -exendina-4; Leu ¹⁴ , Ala ¹⁹ , Phe ²⁵ -exendina-4; exendina-4 (l-30); Leu ¹⁴ -exendina-4 (l-30); Leu ¹⁴ , Phe ²⁵ -exendina-4 (1-30); Leu ¹⁴ , Ala ¹⁹ , Phe ²⁵ -exendina-4 (1-30); exendina-4 (l-28); Leu ¹⁴ -exendina-4 (1-28); Leu ¹⁴ , Phe ²⁵ -exendina-4 (l-28); Leu ¹⁴ , Ala ¹ 9, Phe ²⁵ -exendina-4 (1-28); Leu ¹⁴ , Lys ^{17 <20} , Ala ¹⁹ , Glu ²¹ , Phe ²⁵ , Gln ²⁸ -exendina-4; Leu ¹⁴ , Lys ^{17 <20} , Ala ¹⁹ , Glu ²¹ , Gln ²⁸ -exendin-4; octylGly 14, Gln 28 -exendina-4; Leu 14, Gln 28, octylGly 34 -exendina-4; Phe ⁴ , Leu ¹⁴ , Gln ²⁸ , Lys ³³ , Glu ³⁴ , Ile ^{35 '36} , Ser ³⁷ -exendina-4 (1-37); Phe ⁴ Leu ¹⁴ Lys ^{17 '20} , Ala ¹⁹ , Glu ²¹ , Gln ²⁸ -exendina-4; Val ¹¹ , Ile ¹³ , Leu ¹⁴ , Ala ¹⁶ , Lys ²¹ , Phe ²⁵ -exendina-4; exendin- 4- Lys ⁴⁰ ; lixisenatide; CJC-1134; [N ^e - (17-carboxyheptadecanoic acid) Lys ²⁰ ] exendin-4- H ₂ ; [N ^e - (17-carboxyheptadecanoic acid) Lys ³² ] exendin-4- H ₂ ; [deamino-His ¹ , N ^e - (17-carboxyheptadecanoyl) Lys ²⁰ ] exendin-4- H ₂ ; [Argl ^{2 '27} , NLe ¹⁴ , N ^e - (17-carboxyheptadecanoyl) Lys ³² ] exendin-4- H ₂ ; [N ^e - (19-carboxynonadecanoylamino) Lys ²⁰ ] exendin-4- H ₂ ; [N ^e - (15-carboxypentadecanoylamino) Lys ²⁰ ] - exendin-4- ² ; [N ^e - (13-carboxytridecanoylamino) Lys ²⁰ ] exendin-4- H ₂ ; [N ^e - (11-carboxy-undecanoylamino) Lys ²⁰ ] exendin-4- H ₂ ; exendina-4-Lys ⁴⁰ (e-MPA) - H ₂ ; exendina-4-Lys ⁴⁰ (e-AEEAAEEA-MPA) - H ₂ ; exendina-4-Lys ⁴⁰ (e-AEEA-MPA) - H2; exendina-4-Lys40 (e-MPA) -albumin; GLP-1 (7-37); GLP-1 (7-36) - H ₂ ; LY2428757; desamino-His ⁷ , Arg ²⁶ , Lys ³⁴ (N ^e - (Y-Glu (Na-hexadecanoyl))) - GLP-1 (7-37); disdain His ⁷ , Arg ²⁶ , Lys ³⁸ (N ^e- octanoyl) -GLP-1 (7-37); Arg ^{26 '34,} Lys ³⁸ (N 8 (or carboxypentadecanoyl)) - GLP-l (7-38); Arg ^{26 '34,} Lys ³⁶ (Ns - (y-Glu (N-ahexadecanoil))) - GLP-l (7-36); Aib ^{8 '35} , Arg ²⁶ ' ³⁴ , Phe ³¹ -GLP-1 (7-36);

HXaa ₈ EGTFTSDVSSYLEXaa ₂₂ Xaa ₂₃ A AKEFIXaa ₃ oWLXaa33Xaa ₃ 4G Xaa ₃₆ Xaa ₃₇ ; where Xaa ₈ is A, V, or G; Xaa ₂₂ is G, K, or E; Xaa ₂₃ is Q or K; Xaa ₃₀ is A or E; Xaa ₃₃ is V or K; Xaa ₃₄ is K, N, or R; Xaa ₃₆ is R or G; and Xaa ₃₇ is G, H, P, or is not present; Arg34-GLP-1 (7-37); Glu30-GLP-1 (7-37); Lys22-GLP-l (7-37); Gly ^{8 '36} , Glu ²² -GLP-1 (7-37); Waltz, Glu22, G116 -GLP-1 (7-37); Gll-36, Glu22, Lys33, Asn34-GLP-l (7-37); Val ⁸ , Glu ²² , Lys ³³ , Asn ³⁴ , Gly ³⁶ -GLP-1 (7-37); Gly ³⁶ , Glu ²² , Pro ³⁷ -GLP-1 (7-37); Val ⁸ , Glu ²² , Gly ³⁶ Pro ³⁷ -GLP-1 (7-37); Gly ^{8 '36} , Glu ²² , Lys ³³ , Asn ³⁴ , Pro ³⁷ -GLP-1 (7-37); Val ⁸ , Glu ²² , Lys ³³ , Asn ³⁴ , Gly ³⁶ , Pro ³⁷ -GLP-1 (7-37); Gly ^{8 '36} , Glu ²² -GLP-1 (7-36); Val ⁸ , Glu ²² , Gly ³⁶ -GLP-1 (7-36); Val ⁸ , Glu ²² , Asn ³⁴ , Gly ³⁶ -GLP-1 (7-36); Gly ^{8 '36} , Glu ²² , Asn ³⁴ -GLP-1 (7-36) or any of the above linked to the constant region of an immunoglobulin.

 The term "liraglutide" refers to a GLP-1 analog peptide that is used for the treatment of type 2 diabetes and is marketed under the name Victoza. It has a sequence identity with GLP-1 of 97%, but has been modified in order to protect it from degradation by DPP-4 (dipeptidyl peptidase-4), which has a much longer half-life than GLP-1 and its effect is long lasting. The chemical formula of liraglutide is

-His-Ala-Glu-eiy-Thr-Phe-Thr-Ser-Asp-Vai-Ser Iy-Gly-Leu-Tyr-Ser

and Arg Gly OH

In another possible embodiment, the GLP-1 receptor agonist is Exenatide. The term "Exenatide", as used in the present invention, refers to a GLP-1 analog that binds to its receptor mimicking its action. It has a sequence identity with GLP-1 of 53%. Exenatide is a drug used in the treatment of type 2 diabetes that is marketed under the name of Byetta or Bydureon, a modification of exenatide that requires only a weekly administration in diabetic patients.

 "PPARa" (Peroxisome proliferator-activated alpha receptor) means the protein defined by the sequence described in the UniProt database by the accession number Q07869. PPARa is a transcription factor activated by ligand, implicated in fatty acid oxidation and lipid metabolism. PPARa is expressed in tissues that have a high rate of catabolism of fatty acids, such as liver, heart, skeletal muscle, kidney and adipose tissue, and regulates the expression of genes critical for lipid and lipoprotein metabolism.

 PPARa agonists suitable for use in the present invention include compounds capable of activating the transcription of a reporter gene that is operably linked to a PPARa response region as described in Fukuen et al. (J.Biol.Chem., 2005, 280: 23653-23659) or compounds capable of inducing preadipocyte differentiation in the presence of insulin, as described in Usui et al, Jpn. Chem. Pharm. Bull. (2007) 55: 1053-1059.

In a preferred embodiment, the PPARa agonist is a natural ligand such as oleoylethanolamide. In another possible embodiment, the PPARa agonist is an ethanolamide from among those listed: oleoylethanolamide, palmitoylethanolamide, elaidoylethanolamide or any combination thereof. In another possible embodiment, the PPARa agonist is a compound of those listed below: GW409544, LY-518674, LY- 510929, TZD18, LTB4, LY-465608, pyrinic acid, fatty acids, ragaglitazar, AD - 5061, phenobribic acid, GW7647, GW9578, TAK-559, KRP-297 / MK-0767, eicosatetraenoic acid, farglitazar, reglitazar, DRF 2519, pristanic acid, bezafibrate, clofibrate, 8 S-hydroxy eicosatetraenoic acid, GW- 220, pterostilbene, tetradecylglycidic acid, ortiltiopropiónico acid, Wy14643, ciprofibrate, gemfibrozil, muraglitazar, tesaglitazar, eicosanoids, GW0742X, GW2433, GW9578, GW0742, L- 783483, GW501516, retinoic acid, L-796449, L-165461, L-165041 , SB-219994, LY- 510929, 10 AD-5061, L-764406, GW0072, nTzDpa, troglitazone, LY-465608, pioglitazone, SB-219993, 5-aminosalicyclic acid, GW1929, L-796449, GW7845, acid 2- cyano-3, 12-dioxooleana-l, 9-dien-28-oico, L-783483, L-165461, AD5075, fluorenylmethoxycarbonyl-L-leucine, CS-045, in domethacin, rosiglitazone, SB-236636, GW2331, PAT5A, MCC555, linoleic acid, bisphenol A diglycidyl ether, GW409544, GW9578, TAK-559, reglitazar, W9578, ciglitazone, DRF2519, LG 10074, ibuprofen, diclofenac, fenofibrate, pharmaceutically acceptable salts.

 "Aciletanolamides" are fatty acid amides that act as endogenous mediators of a lipid nature.

 The term "oleoylethanolamide", as used in the present invention, refers to an acylethanolamide whose structure is

It is generated naturally by regulating intake and body weight gain in all vertebrates. It is an endogenous PPARa ligand that regulates its activity to stimulate lipolysis.

 The term "palmitoylethanolamide" refers to the compound whose structure is

In a preferred embodiment, the compositions or kits of parts of the invention comprise a beta-3 adrenergic receptor agonist and a GLP-1 receptor agonist, preferably CL-316,243 and liraglutide. In another preferred embodiment, the compositions or kits of parts of the invention comprise a beta-3 adrenergic receptor agonist and a PPARa agonist, preferably CL-316,243 and oleoylethanolamide. In another embodiment, the compositions or kits-parts of the invention comprise a beta-3 adrenergic receptor agonist, a GLP-1 receptor agonist and a PPARa agonist, preferably CL-316,243, liraglutide and oleoylethanolamide. In a preferred embodiment, the composition or the kit of parts comprises another active ingredient. In a preferred form, said other active ingredient is an antidiabetic compound. Anti-diabetic compounds that can be used in the context of the present invention include compounds that induce the anti-glycemic activity of insulin or sensitizers of insulin activity and include, without limitation, insulin secretagogues such as sulfonylureas (tolbutamide, chlorpropamide , glipicide, glibenclamide, glylicazide, glipentide, glimepiride, libenclamide, glipizide, gliquidone, glisentide, glimepride and the like) and metiglinides (repaglinide, nateglinide, mitiglinide and the like), hepatic and glucose production agents in particular (biguan, bigu , metformin and buformide), agents that cause the decrease of carbohydrates such as α-glucosidases inhibitors (acarbose, miglitol or voglibose), agents that increase peripheral glucose utilization such as thiazolidinediones (rosglitaszone, pioglitazone and the like), GLP- or a GLP-I mimetic (Byetta-Exanatido, Liraglutinido, CJC-1131 (Conju Chem, Exanatide-LAR (Amylin), BIM-51077, ZP-10, the compounds described in WO 00/07617 and the like), exendin, secretin, DPP-IV inhibitors (sitagliptin, saxagliptin, denagliptin, vildagliptin, ALS-2 -0426, ARI- 2243, BI-A, BI-B, SYR-322, MP-513, DP-893, RO-0730699 and the like), SGLT-2 inhibitors (dapagliflozin, and sergliflozin, AVE2268, T- 1095 and the like) and peptides that cause an increase in insulin production (amlintide, pramlintide, exendin), compounds that have GLP-I activity (glucagon-like peptide 1), protein tyrosine phosphatase IB inhibitors, dipeptidyl peptidase inhibitors , insulin secretagogues; fatty acid oxidation inhibitors, A2 antagonists, c-jun terminal kinase inhibitors, insulin; insulin mimetics, glycogen phosphorylase inhibitors, VPAC2 receptor agonists, glucokinase activators, etc.).

In another preferred embodiment, said other active ingredient is a compound capable of reducing the availability of nutrients in the body, which has an anorexigenic effect or which has an anti-obesity effect. Thus, said active ingredient is selected from the group consisting of amylin, amylin analogue agonists, salmon calcitonin, co-cytokine or an agonist thereof, leptin (OB protein), exendin or an exendin analog, GLP1 or a agonist thereof, PYY polypeptides or agonists thereof, agents that affect neurotransmitters or channels Neural ionic agents such as anti depressants, noradrenaline uptake inhibitors, serotinin 2c receptor agonists, some dopamine antagonists, cannabinoid receptor antagonists 1, agents that modulate the leptin / insulin pathway in the CNS including leptin analogs, promoters of leptin transport or leptin receptor promoters, CNTF, neuropeptide Y, agouti-related peptide antagonists, pro-opiomelanocortin, cocaine, amphetamine, alpha melanocyte stimulating hormone analogs, melanocortin-4 receptor agonists, agents that affect insulin activity / metabolism such as tyrosine phosphatase 1 beta protein inhibitors, peroxy somas 7 proliferator-activated receptor antagonists, bromocriptine, somatostatin agonists, agents that increase resting metabolic rate (agonists of the decoupling protein, thyroid hormone receptor agonists) as well as other agents of different types such as melanin, phytostanol analogs, functional oils, inhibitors of fatty acid synthesis, carboxypeptidase inhibitors, intestinal lipase inhibitors, and the like.

Pharmaceutical and / or cosmetic compositions of the invention

The compositions and kits of parts of the invention may be formulated in the form of pharmaceutical and / or cosmetic compositions, which constitute another aspect of the invention. Pharmaceutical and / or cosmetic compositions useful in the practice of the method of the invention include a therapeutically effective amount of the active agents (the beta-3 adrenergic receptor agonist and the GLP-1 receptor agonist or the PPARa agonist), and a pharmaceutically acceptable carrier. The term "pharmaceutical and / or cosmetic composition" means approved by a regulatory agency of the Federal or state government or included in the US Pharmacopoeia. or other generally recognized pharmacopoeia, for use in animals, and more particularly in humans. The term "pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic compound is administered. Such pharmaceutical carriers may be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin. tub, malt, rice, flour, crete, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dry skim milk, glycerol, propylene, glycol, water, ethanol and the like. The composition, if desired, may also contain smaller amounts of wetting or emulsifying agents, or pH buffering agents. These compositions may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, prolonged release formulations and the like. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. The oral formulation may include standard carriers such as pharmaceutical types of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin.

 The composition can be formulated according to routine procedures such as a pharmaceutical composition adapted for intraperitoneal, intravenous, subcutaneous, intramuscular, oral, nasal, parenteral, topical, transdermal, rectal and human-like administration. When necessary, the composition may also include a solubilizing agent and a local anesthetic such as lidocaine to relieve pain at the injection site. When the composition is to be administered by infiltration, it can be dispensed with an infiltration bottle containing water or pharmaceutical grade saline. When the composition is administered by injection, a vial of water for injection or sterile saline can be provided, so that the ingredients can be mixed before administration.

The therapeutic agents of the compositions of the invention, in particular, the beta-3 adrenergic receptor agonist and the GLP-1 receptor agonist, the PPARa agonist or the combination of both, may be presented as a single formulation (for example , as a tablet or capsule comprising a fixed amount of each of the components) or, conversely, may be presented as separate formulations and then combined for joint, sequential or separate administration. The compositions of the invention also contemplate the formulation as a kit of parts wherein the components are formulated separately but packaged in the same container. The person skilled in the art will appreciate that the formulation of the first and second components of the compositions of the invention they can be similar, that is, formulated in a similar manner (in tablets or tablets), which allows their administration by the same route. In the case where the different components of the invention are formulated separately, the two components can be presented in blister. Each blister contains the medications that have to be consumed over a day. If the medications have to be administered several times a day, the medications corresponding to each administration can be arranged in different sections of the blister, preferably recording in each section of the blister the moment of the day in which they should be administered. Alternatively, the components of the composition of the invention can be formulated differently so that the different components are administered differently. Thus, it is possible that the first component is formulated as a tablet or capsule for oral administration and the second component is formulated for intravenous administration.

 The relationship between the components that are part of the compositions of the invention will depend on the beta-3 adrenergic receptor agonist and the GLP-1 receptor agonist or PPARa agonist used in each particular case, as well as on the desired indication. . Thus, the invention contemplates compositions in which the ratio between the quantities of the two components may range between 1000: 1 and 1: 1000, or between 500: 1 and 1: 500, or between 200: 1 and 1: 200, or between 100: 1 and 1: 100, or between 50: 1 and 1: 50, or between 20: 1 and 1: 20 or between 1: 10 and 10: 1, 5: 1 or 1: 5, or between 2: 1 and 2: 1.

Therapeutic uses of the invention

 In another aspect, the invention relates to a composition or kit of parts according to the invention for use in the treatment or prevention of a metabolic disease. In another aspect, the invention relates to the use of a composition or kit of parts according to the invention for the preparation of a medicament for the treatment or prevention of a metabolic disease. In another aspect, the invention relates to a method for the treatment or prevention of a metabolic disease in a subject comprising the administration to said subject of a composition or kit of parts according to the invention.

The expressions "composition", "parts kit" have been defined previously. The term "metabolic disease", as used in the present invention, refers to all types of disorders in which errors and imbalances in metabolism occur as well as in which metabolic processes occur sub-optimally. The term also refers to disorders that can be treated by modulating the metabolism although the disease itself may not have been caused by a metabolic disorder. In a preferred embodiment, the metabolic disease is selected from the group of obesity, hyperglycemia, insulin resistance, type 2 diabetes and dyslipidemias.

The term "obesity," as used in the present invention, refers to the definition of obesity provided by the WHO based on body mass index (BMI), which consists of the relationship between a person's weight (in kg ) and the square of its height in meters. According to this criterion, a BMI of less than 18.5 kg / m ² is considered as insufficient weight or thinness, a BMI of 18.5-24.9 kg / m ² is considered normal weight, a BMI of 25.0-29.9 kg / m ² is considered as overweight in grade 1, a BMI of 30.0-39.0 kg / m ² is considered obese or overweight in grade 2 and a BMI greater than or equal to 40.0 kg / m ² is considered morbid obesity. Alternatively, there are other methods to define the degree of obesity of an individual such as the waist diameter measured at the midpoint between the lower limit of the ribs and the upper limit of the pelvis (in cm), thickness of the folds of the skin and bioimpedance, based on the principle that lean mass transmits electricity better than fat mass.

The term "hyperglycemia", as used in the present invention, refers to a state in which abnormally high levels of blood glucose appear in relation to baseline fasting levels. Specifically, hyperglycemia is understood when fasting blood glucose levels are consistently higher than 126 mg / dL, postprandial glucose levels are greater than 140 mg / dL and / or venous plasma glucose levels 2 hours after administration of a glucose dose of 1.75 grams per kilogram of body weight is greater than 200 mg / dL. The term "insulin resistance", as used in the present invention, refers to a disorder in which cells do not respond properly to insulin. As a result, the pancreas produces more insulin in response to elevated blood glucose levels. Patients with insulin resistance frequently show high levels of glucose and high levels of circulating insulin. Insulin resistance It is frequently linked to obesity, hypertension and hyperlipidemia. Additionally, insulin resistance appears frequently in patients with type 2 diabetes.

 The term "type 2 diabetes", as used in the present invention, refers to a disease characterized by an inappropriate elevation of blood glucose levels that causes chronic complications due to the involvement of large and small vessels and nerves. The underlying alteration in this disease is the difficulty for the action of insulin (such as a loss of tissue sensitivity to this hormone) that is called insulin resistance and an inadequate secretion of insulin by the cells responsible for its production in the pancreas. In addition to increasing the concentration of glucose, the deficient action of insulin often results in elevated cholesterol and / or triglyceride levels.

 The term "dyslipidemia", as used in the present invention, refers to any pathological condition characterized by an alteration in lipid metabolism, with its consequent alteration of lipid concentrations (cholesterol, triglycerides and the like) and lipoproteins ( high density lipoproteins) in the blood. Dyslipidemia that can be treated with the methods of the present invention include, without limitation, hypercholesterolemia, hypertriglyceridemia, hyperlipoproteinemia type I, Ha, Ilb, III, IV, V, hyperkylroneronemia, combined hyperlipidemia, etc.

 The term "subject", as used in the present invention includes living organisms, such as human beings, female or male, and of any race or age; animals, for example monkeys, cows, sheep, horses, pigs, goats, dogs, cats, mice, rats and transgenic species thereof. In a preferred embodiment, the subject is a human being.

 The different embodiments of the therapeutic method according to the invention in terms of beta-3 adrenergic receptor agonists, GLP-1 receptor agonists and PPARa agonists have been described previously in the context of the compositions of the invention.

The amount of active agents (the beta-3 adrenergic receptor agonist and the GLP-1 receptor agonist or PPARa agonist) that will be effective in the treatment of metabolic disease can be determined by standard clinical techniques based on the present description. . In addition, they can optionally be used in vitro assays to help identify optimal dosing intervals. The precise dose to be used in the formulation will also depend on the route of administration, and the severity of the condition, and should be decided according to the judgment of the doctor and the circumstances of each subject. However, dosage ranges suitable for intravenous administration are generally about 50-5000 micrograms of active compound per kilogram of body weight. Dosage ranges suitable for intranasal administration are generally about 0.01 pg / kg body weight to 1 mg / kg body weight. Effective doses can be extrapolated from dose response curves derived from in vitro or animal model test systems.

 For systemic administration, a therapeutically effective dose can be estimated initially from in vitro assays. For example, a dose can be formulated in animal models to achieve a circulating concentration range that includes the IC50 (a measure of drug efficacy that indicates how much of a substance is necessary to inhibit a given biological process) that has been determined in culture. mobile. Such information can be used to more accurately determine useful doses in humans. Initial dosages can also be estimated from in vivo data, eg, animal models, using techniques that are well known in the state of the art. Someone with normal experience in the art can easily optimize human administration based on data in animals.

 In a preferred embodiment, administration of the first component and the second component in the therapeutic method is carried out so that both components are administered below their individual ED50 or ED10.

 The term "ED50", as used in the present invention, refers to the dose of the active ingredient that produces the therapeutic effects in 50% of patients.

 The term "ED10", as used in the present invention, refers to the dose of the active ingredient that produces the therapeutic effects in 10% of patients.

The ED50 and ED10 values can be calculated in animal models and easily be extrapolated to humans by a person skilled in the art. Therapeutic effect means a significant reduction in food consumption and content total lipid Total lipid content means all body mass that does not consist of bone, muscle, connective tissue and fluids. One method of measuring the total lipid content is the determination of the bioelectrical impedance or bioimpedance, which determines the electrical impedance or the opposition to the flow of the electric current through the body tissues.

Cosmetic uses of the invention

The compositions and kits of parts of the invention are useful both for the treatment of morbid obesity and for the treatment of overweight grade 1 or grade 2, in which case the methods of the invention have a cosmetic purpose. Therefore, in another aspect, the invention relates to a cosmetic method for the treatment of obesity, for the reduction of the total lipid mass and / or for the reduction of the total food consumption of a patient, which comprises administering to a patient a combination or kit of parts according to the invention. In this embodiment, patients who are overweight in the form of fat and who can be treated by the cosmetic method of the present invention are visually identified or because they have a BMI greater than or equal to 25 kg / m ² , preferably between 25 and 30. These individuals are considered as obese who need weight control for cosmetic reasons.

 In another aspect, the invention relates to a cosmetic method for the treatment of cellulite. The term "cel ulitis" applies to changes in the subcutaneous adipose panicle, which occur as a result of an accumulation of hypertrophic adipocytes, and is most commonly found in iaterae areas of the trunk and in the upper thighs.

 The different embodiments of the cosmetic method according to the invention in terms of beta-3 adrenergic receptor agonists, GLP-1 receptor agonists and PPARa agonists have been described previously in the context of the methods of treatment of obesity and other metabolic diseases.

The compositions according to the invention, both for therapeutic and cosmetic use, can be administered chronically, acutely or subchronously. The term "chronic administration", as used in the present invention, refers to a method of administration in which the compound is administered to the patient continuously for extended periods of time in order to maintain the therapeutic effect during said period. . Chronic administration form includes the administration of multiple doses of the compound on a daily basis, twice a day, three times a day or less frequently. Chronic administration can be carried out by several intravenous injections administered periodically over a single day. Alternatively, chronic administration involves administration in the form of a bolus or by continuous transfusion that can be carried out daily, every two days, every 3 to 15 days, every 10 days or more. Typically, chronic administration is maintained for at least one week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 11 weeks, at least 12 weeks, at least 4 months, at least 5 months, at least 6 months, at least 9 months, at least one year, at least 2 years or more.

 The term "acute administration", as used in the present invention, refers to a method of administration in which the patient is exposed to a single dose of the compound or to several doses but for a reduced period of time as per example, 1, 2, 4, 6, 8, 12 or 24 hours or 2 or 3días.

 The term "subchronic administration", as used in the present invention, refers to a method of administration in which the patient is exposed to one or several doses of the compound for a period of time between 3 and 7 days of treatment, such as 3, 4, 5, 6 or 7 days. Typically, subchronic administration is maintained for 6 days.

The person skilled in the art will appreciate that the therapeutically effective amount, and / or formulation of the active compound will be carried out depending on the type of administration. By "therapeutically effective amount," as used herein, is meant the amount of compound that makes it possible to totally or partially relieve the symptoms associated with a metabolic disease or that prevents the progression or worsening of the symptoms or that prevents the onset of disease in a subject at risk of suffering the disease. In the event that a chronic administration of the compound of the invention is desired, it may be administered in a sustained release composition such as those described in US5672659, US5595760, US5821221, US5916883 and W09938536. On the contrary, if acute administration is desired, a treatment with an immediate release form will be preferred. Regardless of the type of administration, the dosage amount and range can be adjusted individually to provide plasma levels of the compounds that are sufficient to maintain the therapeutic effect. Someone with a normal experience in the art will be able to optimize therapeutically effective local dosages without too much experimentation.

 In a preferred embodiment, administration of the first component and the second component in the cosmetic method is carried out so that both components are administered below their individual ED50 or ED10.

 ***

 The invention is described below by the following examples that should be considered as merely illustrative and in no case limiting the scope of the present invention.

EXAMPLES

RNA isolation and analysis by qRT-PCR

RNA was extracted from samples of white adipose tissue (visceral fat) using the Trizol® method, according to the manufacturer's instructions (Gibco BRL Life Technologies, Baltimore, MD, USA). Portions of white adipose tissue tissue (-100 mg) were placed in 1 ml of Trizol reagent (Invitrogen, CA, USA) and homogenized with an IKA-Ultra-Turrax® T8 (IKA-Werke GmbH, Staufen, Germany) . To ensure the purity of the mRNA sequences excluding molecules smaller than 200 nucleotides and proteins, the RNA samples were isolated with the RNAeasy minelute cleanup kit (Qiagen, Hilden, Germany), which included DNase I digestion (DNase set) without RNasa, Qiagen), according to the manufacturer's instructions. The total mRNA concentration was quantified using a spectrophotometer (Nanodrop 1000, Thermo Scientific, Rochester, NY, USA) spectrophotometer to ensure A260 / A280 ratios of 1.8 to 2.0. The reverse transcription reaction was carried out from 1 μm of white adipose tissue mRNA using the Transcriptor reverse transcriptase kit and random hexamic primers (RT Transcriptor, Roche Diagnostic GmbH, Manheim, Germany). Negative controls included reverse transcription reactions that omitted reverse transcriptase. Reverse transcription polymerase chain reaction in quantitative real time (quantitative RT-PCR) was performed using a CFX96TM real-time PCR detection system (Bio-Rad, Hercules, CA, USA) and the FAM dye marker format for TaqMan® gene expression assays (Applied Biosystems). Each reaction was run in duplicate, containing 9 μΐ of diluted cDNA 1/100. The parameters of the cycles were: 50 ° C for 2 minutes to deactivate single and double stranded DNA containing dUTP, 95 ° C for 10 minutes to activate Taq polymerase DNA followed by 40 cycles at 95 ° C for 15 seconds for the fusion of the CDNA and 60 ° C for 1 minute to allow hybridization and extension of the primers in which the fluorescence was acquired. Fusion curve analyzes were performed to ensure that only a single product was amplified. Several maintenance genes were analyzed, selecting the most appropriate ones according to their homogeneity. The absolute values of each sample were normalized with respect to the Gadph maintenance gene. Relative quantification was obtained using the AACt method and normalized with respect to the control group. The primers for the PCR reaction were obtained based on the Applied Biosystems genome database of rat mRNA references (http://bioinfo.appliedbiosystems.com/genome- database / gene-expression.html).

Immunofluorescence

Brown adipose tissue samples were fixed in 4% paraformaldehyde in 0.1 M phosphate buffered saline (PBS, pH 7.4) by immersion and embedded in paraffin. The tissue blocks were cut into sections of 5 μιη of thickness using a Micromomo Microm HM325 (MICROM, Walldorf, Germany). The sections were mounted on glass slides with the positively charged surface (DAKO Real, ref. S2024, Glostrup, Germany). The sections were dewaxed, washed several times with PBS and incubated in 3% hydrogen peroxide in PBS for 20 minutes in the dark at room temperature to inactivate the endogenous peroxidase. After 3 washes in PBS for 5 minutes, antigen recovery was achieved by incubation in sodium citrate (pH 6) for 15 minutes at 95 ° C. A bottom blocking solution containing 10% donkey serum, 0.3% triton X-100 and 0.1% sodium azide was used to incubate the sections for 1 hour, followed by overnight incubation. at room temperature with the primary polyclonal goat antibody UCPl (diluted 1: 50, Santa Cruz, catalog number sc-14720). The sections were washed three times with PBS, incubated for 2 hours in an Alexa Fluor® donkey IgG goat IgG conjugated with Cy ™ 3 (Invitrogen-Life Technologies, catalog number Al 1057, Paisley, UK) diluted 1: 200 and washed in phosphate buffer.

 High resolution digital photomicrographs of brown adipose tissue were taken under the same light and brightness / contrast conditions using an Olympus BX41 microscope equipped with a 40x lens, an Olympus DP70 digital camera (Olympus Europa GmbH, Hamburg, Germany) and a system of Olympus X-cite fluorescence (X-cite 120Q series, Olympus). Immunofluorescence quantification was carried out by measuring the densitometry of the images obtained from replicates and samples: two replicates per sample, four samples per group and four groups, using ImageJ l analysis software, 38x (National Institute of Health, Bethesda , Maryland, USA). Statistic analysis

All data are represented as mean ± SEM (standard error of the mean) of at least eight determinations per experimental group. The Kolmogorov-Smirnov normality tests indicated that all data followed a Gaussian distribution (P> 0, 1), so that a parametric statistical test was selected. Differences between treatments were analyzed by one-way VA ANO for repeated measures, followed by the subsequent Bonferroni test for multiple comparisons. The level of statistical significance was adjusted to P values of less than 0.05. Example 1: Effect of the beta-3 adrenergic receptor agonist CL316243 on the regulation of intake and circulating triglyceride levels. The adrenergic beta-3 receptor agonist CL316243 (Fig. 1A) was given to different doses (0.1 mg / kg, 0.5 mg / kg and 1 mg / kg) to deprived rats of food by intraperitoneal injection, and analyzed the effect on intake 30 min, 60 min, lh, 2h, 4h, 6h, 8h, 12h and 24h after injection (Fig. IB). CL316243 significantly reduces food intake at all doses used. Likewise, circulating triglyceride levels were measured in rats treated with different concentrations of CL316243, with a reduction in triglyceride concentration dependent on the concentration of CL316243 administered (Fig. IB).

Example 2: Effect of the combined treatment of the adrenergic beta-3 receptor agonist CL316243 and the GP-1 receptor agonist liraglutide.

 A subchronic treatment was administered to the rats, consisting of a daily intraperitoneal injection of CL316243, two daily subcutaneous injections of Liraglutida or a combination of both (CL316243 and Liraglutide simultaneously) for 6 days, and the effect of inhibition treatment was analyzed. of intake (Fig. 2A), amount of total lipid mass (Fig. 2B) and circulating triglyceride levels (Fig. 2C). The combination of CL316243 and Liraglutida exerts a clear synergistic effect in reducing the total lipid mass of the animals.

Example 3: Effect of the combined treatment of the adrenergic beta-3 receptor agonist CL316243 and the PPARa agonist oleoyletanolamide.

A subchronic treatment was administered to the rats, consisting of a 6-day daily intraperitoneal injection of CL316243 or oleoylethanolamide, or a combination of both (CL316243 and oleoylethanolamide simultaneously), and the effect of the treatment as regards intake inhibition was analyzed ( Fig. 3 A), amount of total lipid mass (Fig. 3B) and circulating triglyceride levels (Fig. 3C). The combination of CL316243 and oleoyletanolamide exerts a clear synergistic effect in reducing the total lipid mass of the animals. Example 4: Effect of treatment with CL316243 and oleoylethanolamide on the expression of UCPl

 The combinatorial therapy of CL316243 and oleoyletanolamide increases the expression of the gene that encodes the decoupling protein 1 (UCPl), the main responsible for triggering thermogenesis. This effect is observed in both white adipose tissue (Fig. 4) and brown adipose tissue (Fig. 5 A), and is associated with an increase in the expression of the UCPl protein, as observed by immunohistochemistry (Fig. 5B). These results indicate that the main effect of combinatorial therapy to reduce body fat is derived from an increase in the mobilization of fatty acids coupled with an increase in thermogenic responses.

Claims

A composition or kit of parts comprising, together or separately, a first component and a second component, wherein the first component is a beta-3 adrenergic receptor agonist and the second component is selected from the group consisting of a receptor agonist of GLP-1, a PPARa agonist and a combination of both.

2. The composition or the kit of parts according to claim 1, wherein the agonist of the rheumatic beta-3 receptor is a compound with the formula

 pharmaceutically acceptable derivative salt, where

R2 is

R ₃ can also be (CH

COR ₄ o

 R4 is a hydroxy, alkoxy, amino, alkylamino or dialkylamino

R ₅ is hydrogen, fluorine, chlorine, bromine, iodine, -CN, CF ₃ , short chain alkyl, short chain alkoxy, cycloalkyl or aryl;

R _Ó is short chain alkyl, cycloalkyl or aryl;

R ₇ , R ₇ ', R ₈ and R ₈ ' can independently hydrogen, short chain alkyl or R ₇ and R ₈ can be together CH ₂ -CH ₂ ;

Z is hydrogen or

m is an integer between 1 and 2;

n is zero or an integer between 1 and 6,

p is an integer between 1 and 5

The composition or kit of parts according to claim 2, wherein the beta-3 adrenergic receptor agonist is (R, R) -5- [2- [2- (3-Chlorophenyl) -2-hydroxyethylamino] propyl] -l,

3-benzodioxol-2,2-dicarboxylic, whose formula is

4. The composition or kit of parts according to any one of claims 1 to 3, wherein the GLP-1 agonist is liraglutide.

5. The composition or kit of parts according to any one of claims 1 to 4, wherein the PPARa agonist is selected from the list consisting of oleoylethanolamide, palmitoylethanolamide, elaidoylethanolamide, or any combination thereof.

6. The composition or kit of parts according to any one of claims 1 to 5, wherein the PPARa agonist is oleoylethanolamide.

7. The composition or kit of parts according to any one of claims 1 to 6 which further comprises another active ingredient.

8. A pharmaceutical or cosmetic composition comprising a composition or a kit of parts according to any one of claims 1 to 7 and a pharmaceutical and / or cosmetically acceptable carrier.

9. Use of a composition or a kit of parts according to any of claims 1 to 7 for the preparation of a medicament.

10. Use of a composition or kit of parts according to any one of claims 1 to 7 for the preparation of a medicament for the treatment or prevention of a metabolic disease.

11. Use according to claim 10 wherein the metabolic disease is selected from the group of obesity, dyslipidemia, insulin resistance, hyperglycemia and type 2 diabetes.

12. Use according to claim 10 or 11, wherein said composition or kit of parts is administered subchronously.

13. Use according to any of claims 10 to 12, wherein said composition or the kit of parts is administered orally, rectally, parenterally, intraperitoneally, topically or transdermally.

14. Use according to any of claims 10 to 13 wherein the first component and the second component are administered below their individual ED50 or ED10.

Cosmetic method for the treatment or prevention of obesity and / or cellulite or to reduce the total lipid mass, or to reduce food consumption by a mammal, which comprises administering to a patient a pharmacologically active amount of a composition or kit of parts according to any one of claims 1 to 7.

Method according to claim 15 wherein the treatment is carried out subchronously.

17. Method according to claim 15 or 16 wherein the composition is administered by intraperitoneal injection or by transdermal route.

18. Method according to any of claims 14 to 16, wherein the first component and the second component are administered below their individual ED50 or ED10.