WO2011080276A1

WO2011080276A1 - Pharmaceutical combinations comprising a dpp-4 inhibitor and a 1,3-diphenylprop-2-en-1-one derivative

Info

Publication number: WO2011080276A1
Application number: PCT/EP2010/070808
Authority: WO
Inventors: Rémy HANF; Anne Rubenstrunk; Robert Walczak
Original assignee: Genfit SA
Current assignee: Genfit SA
Priority date: 2009-12-29
Filing date: 2010-12-28
Publication date: 2011-07-07
Anticipated expiration: 2012-06-29

Abstract

The present invention provides novel combination products comprising 1,3-diphenylprop-2-en-1-one derivatives of General Formula (I) and a DPP-4 Inhibitor and pharmaceutical compositions comprising the same for treating disorders that require preserving physiological levels of endogenous GLP-1. 1,3-diphenylprop-2-en-1-one derivative of General Formula (I) potentiates DPP-4 Inhibitor effects and their combination can be used in methods for treating a condition mediated by GLP-1, such as metabolic disorders.

Description

PHARMACEUTICAL COMBINATIONS COMPRISING A DPP-4 INHIBITOR AND A 1 ,3- DIPHENYLPROP-2-EN-1 -ONE DERIVATIVE

TECHNICAL FIELD

The invention relates to the use of combinations of compounds in the preparation of pharmaceutical compositions and methods for the treatment of metabolic or neurodegenerative disorders.

BACKGROUND

Metabolism and energy homeostasis is regulated by the brain in response to signals from the adipose tissue, pancreas and the gastrointestinal tract. In particular, Glucagon- Like peptide 1 (GLP-1 ) is a peptide that is released by L cells (a specific cell type in the gastrointestinal tract) following food intake and tissue specific cleavage of proglucagon. As recently reviewed (Wajchenberg B, 2007; Campbell R, 2009a; Reimann F et al. 2006; Ranganath L, 2008), GLP-1 acts as a gut-derived incretin hormone that inhibits gastric emptying and further nutrient ingestion, potentiating insulin biosynthesis and suppressing glucagon secretion. GLP-1 acts on pancreatic beta-cells, inducing their proliferation and expansion, promoting islet neogenesis, and inhibiting apoptosis. Extra-pancreatic effects of GLP-1 have also been reported, suggesting in particular that this peptide can have neuroprotective and neuroproliferative properties.

GLP-1 regulatory effects on glucose homeostasis have been considered of major interest for developing new therapies of disorders that requires lowering of plasma glucose concentration and the overall improvement of glycemic control such as for the treatment of type-2 diabetes. However, GLP-1 effects are hampered by its very short half- life in circulation since it is rapidly broken and inactivated by dipeptidyl peptidase-4 (DPP- 4), a member of the S9b serine protease family that cleaves a number of regulatory factors for metabolism. In fact, DPP-4 defective transgenic mice are refractory to the development of obesity and hyperinsulinemia (Conarello S et al., 2003; Drucker D, 2007).

Current therapeutic approaches for enhancing GLP-1 action target either GLP-1 receptor (by using degradation-resistant GLP-1 receptor agonists that mimic endogenous GLP-1 ) or DPP-4 enzymatic activity (by using small molecules as DPP-4 Inhibitors that slow down the otherwise rapid degradation of endogenous GLP-1 ). Both categories of compounds provide positive effects on the treatment of type 2 diabetes when compared to other treatments that target pancreatic, intestinal, liver, adipose tissue functions and metabolism and that have been disappointing in producing a long-lasting impact on glycemic control (Campbell R, 2009b; Pratley R and Salsali A, 2007).

GLP-1 receptor agonists that administered by sub-cutaneous injection are on the market and long acting, orally administered variants are currently in (pre-)clinical development. They have shown good efficacy on type 2 diabetes and show some improvements on weight loss, systolic blood pressure, beta cell function together with some beneficial effects on cardiovascular risk factors (Hansen K et al., 2009) when compared to classical anti-diabetic medications. In parallel, various classes of structurally different DPP-4 Inhibitors are currently under development and some of them, such as Sitagliptin and Vildagliptin, have been already launched. They have good therapeutic effectiveness on the secretion of insulin or the inhibition of glucagon release after a meal as a once daily oral monotherapy and as add-on therapy with current other anti-diabetic drugs with good safety profile and high tolerability (Gupta R et al., 2009; White J, 2009).

Comparative studies have been performed on the effects and the mechanism of action of GLP-1 receptor agonists and DPP-4 Inhibitors and on the different advantages of either increasing GLP-1 levels or preserving physiologic levels of endogenous GLP-1 on weight loss, pancreatic functions, glucose-lowering effects, undesired effects, pharmacokinetics/pharmacodynamics, immunogenicity, and/or route of administrations (Neumiller J, 2009; Wajchenberg B, 2007; Drucker D, 2006). These studies prompted different proposals on strategies for the medical management of metabolic disorders by adapting the treatment with the specific clinical status (Inzucchi S and McGuire D, 2008).

In particular, the combination of two or more compounds having different mechanisms of action, regimens, and effects is considered as a promising strategy for the treatment of diabetes. This therapeutic approach may not only provide the simple additive effects of the compounds but also potentiate the activity and improve other major clinical end points of one compound by means of another compound for treating type 2 diabetes, in particular by using compounds that are orally administered (Campbell I, 2009; Ahren B, 2009; Bolen S et al., 2007).

The additive glucose lowering effects of different classes of compounds when combined with either GLP-1 receptor agonists or DPP-4 Inhibitors have been suggested and, in a few cases, demonstrated in preclinical and clinical studies after chronic coadministration. Apart from early generic disclosures (such as in WO2001/52825), specific combinations include compounds that are activators of PPARgamma and/or PPARalpha (Brand C et al., 2009; Seufert J, 2009; Rosenstock J et al., 2007; WO2009/1 1 1078; WO2006/000567; WO2006/047248; WO2004/052362), agonists of molecules such as BRS-3 (WO2007/027225), and GPR1 19 (WO2007/120702), or antagonists of molecules such as SGLT2 (WO2009/022010) or CB1 receptor (WO2006/1 19260). However, the need for novel therapeutic options for the medical management of disorders requiring the potentiation of activity of DPP-4 Inhibitors is still clear and urgent.

SUMMARY OF INVENTION

The present invention is based on the surprising finding that the chronic administration of a 1 ,3-diphenylprop-2-en-1 -one derivative provides, when it is associated with an acute administration of a DPP-4 Inhibitor, a statistically relevant increase of GLP-1 concentration in the plasma following glucose administration. The invention provides novel combination products comprising 1 ,3-diphenylprop-2- en-1 -one derivatives (said derivatives being elsewhere also referred to as the "Compounds of General Formula (I)) and a DPP-4 (Dipeptidyl Peptidase 4) Inhibitor, or pharmaceutical compositions comprising the same compounds, for use in a method for the treatment of disorders that require preserving physiological levels of endogenous GLP-1 (Glucagon-Like Peptide 1 ) by potentiating the effects of DPP-4 Inhibitor, in particular for treating metabolic disorders. The present invention also provides novel therapeutic uses and methods of administration of Compounds of General Formula (I) in combination with a DPP-4 Inhibitor.

Further objects of the present invention, including pharmaceutical compositions and methods of administration, are provided in the Detailed Description.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel combination products, pharmaceutical compositions, and methods of administration comprising specific 1 ,3-diphenylprop-2-en-1 - one derivatives that are substituted on both phenyl groups for treating conditions mediated by GLP-1 , such compounds being capable of increasing in a surprising manner GLP-1 level, and thus potentiating the efficacy of a DPP-4 Inhibitor for treating disorders that require preserving physiologic levels of endogenous GLP-1 levels in the blood. The compounds that are defined by means of General Formula (I) can improve the therapeutic efficacy of a DPP-4 Inhibitor through pharmaceutical combinations that were neither disclosed nor suggested in the prior art. The term "DPP-4 Inhibitor" refers to any compound that is recognized as being capable to avoid, reverse or prevent the modification of GLP-1 in the plasma due to DPP-4 enzymatic activity. The DPP-4 Inhibitor exerts this action by binding DPP-4 and inhibiting DPP-4 dipeptidyl peptidase activity with sufficient selectivity for DPP-4 over closely related peptidases, such as one dipeptidyl peptidase-2, -8, or -9 (DPP-2, DPP-8, or DPP-9). The literature provides several lists of compounds belonging to different families of chemically distinct structures that are characterized as DPP-4 Inhibitors and that are at different level of (pre)-clinical development (Levien T and Baker D, 2009; Havale S and Pal M, 2009). A non-exhaustive list of DPP-4 Inhibitors comprises the molecules that are identified in the literature as Sitagliptin, Vildagliptin, Saxagliptin, Alogliptin, Carmegliptin, Gosogliptin, Tenegliptin, Retagliptin, Linagliptin, Melogliptin, DA-1229, ABN-279, AMG- 222, PSN-9301 , P32/98, LC-150444, LC-150033, TAK-100, SYR-322, SYR-472/TAK-472, CWP-0403, KRP-104, Sulphostin, or their pharmaceutically acceptable salts. According to a particular embodiment of the invention, the DPP-4 Inhibitor is selected in the group consisting of Sitagliptin, Vildagliptin, Saxagliptin, Alogliptin and the pharmaceutically acceptable salts thereof.

The expression "a condition mediated by GLP-1 " refers to any disorder that is ameliorated by increasing GLP-1 level (or preserving physiological levels of endogenous GLP-1 ) in the blood. This increase should be sufficient to lower a blood glucose level, and in general for improving glycemic control and/or for reducing of fasting or postprandial plasma glucose in subjects affected by metabolic disorders such as type 2 diabetes.

In particular, the condition mediated by GLP-1 is selected from a metabolic disorder (such as diabetes, obesity, a disorder related to diabetes, or a disorder related to pancreatic beta-cells number and/or activity), or a neurodegenerative disorder. Preferred metabolic disorders are type 2 diabetes mellitus, type 1 diabetes mellitus, hyperglycemia, prediabetes, impaired glucose tolerance, impaired fasting blood glucose, insulin resistance, hypertriglyceridemia, hypercholesterolemia, dyslipidemia, hyperlipidemia, atherosclerosis, hypertension, sleep apnea, polycystic ovarian syndrome, and/or from metabolic syndrome, obesity (indicated as a body mass index of 30.0 or greater, in accordance with the WHO classifications of weight, but including overweight, visceral obesity, abdominal obesity, and in general the reduction, or the prevention of the increase of body weight), or a disorder related to diabetes. A non-exhaustive list of diseases related to diabetes includes complications of type 2 diabetes mellitus such as cataracts and micro- and macrovascular diseases, diabetic nephropathy, diabetic retinopathy, diabetic neuropathy, enteroendocrine cell insufficiency, glucosuria, metabolic acidosis, tissue ischaemia, arteriosclerosis, diabetic coronary artery disease, diabetic cerebrovascular disease, diabetic peripheral vascular disease, myocardial infarction, stroke and peripheral arterial occlusive disease.

The expression "disorders related to pancreatic beta-cells number and/or activity" refers to disorders involving the degeneration of pancreatic beta-cells and/or the decline of the functionality of pancreatic beta-cells (in particular, insulin secretion) by increasing the number and/or the size of beta-cells in a subject, which is a major biological activity of GLP-1 (Buteau J, 2008).

The combinations and the methods of the invention can be also useful or applicable for the treatment of neurodegenerative disorders. The neuroprotective and neuroproliferative effects of GLP-1 may explain the interaction between diabetes and presymptomatic Alzheimer's disease (Burdo J et al., 2009) and experimental data on models for brain insulin signalling and Alzheimer's disease (de la Monte S and Wands J, 2008). The neurodegenerative disease can be selected from excitotoxic brain damage caused by severe epileptic seizures, Alzheimer's disease, Parkinson's disease, Huntingdon's disease, motor-neuron disease, learning and/or memory impairment, traumatic brain injury, spinal cord injury, and peripheral neuropathy.

The term "treatment" or "treating" refers to therapy, prevention, and prophylaxis of a disorder, in particular of a condition mediated by GLP-1 , in a subject in need thereof. The treatment involves the administration of a combination or of a pharmaceutical composition to patients having a declared disorder to prevent, cure, delay, reverse, or slow down the progress, improving thereby the condition of patients. A treatment may be also administered to either healthy subjects that are at risk of developing a condition mediated by GLP-1 , or a subject with insufficient glycemic control despite monotherapy with a DPP- 4 Inhibitor.

The term "subject" refers to a mammal and more particularly a human. The subjects to be treated according to the invention can be appropriately selected on the basis of several criteria associated to a condition mediated by GLP-1 such as previous and/or present drug treatments, associated pathologies, genotype, exposure to risk factors, as well as any other relevant biomarker that can be evaluated by means of immunological, biochemical, or enzymatic method (Pfijtzner A et al., 2008). In particular, the subject is a patient at risk of developing a condition mediated by GLP-1 , and/or who shows one, two or more of the following conditions: a fasting blood glucose or serum glucose concentration greater than 1 10 mg/dL, a postprandial plasma glucose equal to or greater than 140 mg/dL; an HbAlc value greater than 6.0; triglyceride blood level≥ 150 mg/dL; HDL-cholesterol blood level < 40 mg/dL in female and < 50 mg/dL in male patients.

The compounds to be used and administered in combination products and/or comprised in pharmaceutical compositions together with a DPP-4 Inhibitor for use in a method for the treatment of a condition mediated by Glucagon-Like Peptide 1 according to the invention have the following General Formula (I):

in which:

X1 represents a halogen, a R1 or G1 -R1 group;

A represents a CH=CH or a CH2-CH2 group;

X2 represents a G2-R2 group;

G1 and G2, identical or different, represent an atom of oxygen or sulphur; R1 represents a hydrogen atom, an unsubstituted alkyl group, a cycloalkyl group, an aryl group or an alkyl group that is substituted by one or more halogen atoms, alkoxy, alkylthio, cycloalkyl, cycloalkylthio, or heterocyclic groups;

R2 represents an alkyl group substituted by at least a -COOR3 group, wherein R3 represents a hydrogen atom, or an alkyl group that is substituted or not by one or more halogen atoms, cycloalkyl groups, or heterocyclic groups;

R4 and R5, identical or different, representing an alkyl group that is substituted or not by one or more halogen atoms, cycloalkyl groups, or heterocyclic groups.

In a particular embodiment, compounds of General Formula (I) present at least an alkyloxy group or an alkylthio group in X1 and X2 positions (i.e., at least one of X1 and X2 represents an alkyloxy group or an alkylthio group). Moreover, the derivatives can be in the form of substituted 1 ,3-diphenylpropanones that are obtained by reduction of the corresponding 1 ,3-diphenylprop-2-en-1 -one derivatives, that is, the A group CH=CH is replaced by CH2-CH2.

In a particular embodiment, X1 is a G1 -R1 group, and more preferably G1 is a sulphur atom and R1 is a linear or branched alkyl group that is substituted or not by one or more halogen atoms, cycloalkyl groups, heterocyclic groups. Even more preferably, X1 is an alkylthio group that comprises an alkyl group that is linear or branched, having from one to seven carbon atoms that is substituted or not by one or more halogen atoms. In a preferred embodiment, X1 is a methylthio group.

In a particular embodiment, X2 is a G2-R2 group, wherein G2 is an oxygen atom and R2 is an alkyl group substituted by a -COOR3 group, wherein R3 represents a hydrogen atom or an unsubstituted linear or branched alkyl group having from one to seven carbon atoms, and more preferably from one to four carbon atoms

Furthermore, R4 and R5, identical or different, can be unsubstituted linear branched, alkyl groups having from one to seven carbon atoms, and more preferably from one to four carbon atoms. In a preferred embodiment, both R4 and R5 represent methyl groups. In the context of the present invention, the term "alkyl" refers to a saturated hydrocarbon radical that is linear or branched, having preferably from one to twenty-four, and even more preferably from one to seven carbon atoms, such as methyl, ethyl, n- propyl, isopropyl, n-butyl, isobutyl, tertiobutyl, sec-butyl, pentyl, neopentyl, or n-hexyl.

The term "alkyloxy" refers to an alkyl group as defined above that is linked to the remainder of the compound by an oxygen atom (ether bond).

The term "alkylthio" refers to an alkyl group as defined above that is linked to the remainder of the compound by a sulfur atom (thioether bond).

The term "cycloalkyl" designates an alkyl group that forms one cycle having preferably from three to fourteen carbon atoms, and more preferably three to eight carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. The cycloalkyl group can be substituted or not by one or more halogen atoms.

The term "cycloalkylthio" refers to a cycloalkyl group as defined above that is linked to the remainder of the compound by a sulfur atom (thioether bond).

The term "aryl" designates an aromatic group, substituted or not having preferably from six to fourteen carbon atoms, such as phenyl, a-naphtyl, b-naphtyl, biphenyl, anthracenyl.

The term "heterocyclic" refers to a heterocycloalkyl group or a heteroaryl group.

The term "heterocycloalkyl" group refers to a cycloalkyl as indicated above that further comprises one or several heteroatoms selected among nitrogen, oxygen or sulfur. They generally comprise from four to fourteen carbon atoms, such as morpholinyl, piperidinyl, tetrahydropyranyl, dithiolanyl.

The term "heteroaryl" group refers to an aryl group as indicated above that further comprises one or several heteroatoms selected among nitrogen, oxygen or sulfur. They generally comprise from four to fourteen carbon atoms, such as furanyl, thiophenyl, pyridinyl, pyrimidinyl, quinoleinyl, isoquinoleinyl.

By halogen atom, an atom of bromine, chlorine, fluorine or iodine is understood. Different families of 1 ,3-diphenylprop-2-en-1 -one derivatives and 1 ,3- diphenylpropanones that are substituted on both phenyl groups can be found in the prior art (WO2003/037315, WO2001/0461 10, JP2006-303800, JP04-202129). However, none of these documents shows that specific effects on the plasma GLP-1 levels are associated to compounds as defined in General Formula (I).

The structure, synthesis, and some activities of compounds that are encompassed by General Formula (I) have been disclosed in a series of patent applications (WO2004/005243, WO2004/005233, WO2005/005369, US20070032543, WO2005/073184, WO2007/147879, and WO2007/147880). The use of such compounds in combination with a DPP-4 Inhibitor in methods for treating a condition mediated by GLP-1 has neither been disclosed nor suggested by any of these documents.

Specific 1 ,3-diphenylprop-2-en-1 -one derivatives of General Formula (I) that can be used and administered in combination products and/or comprised in compositions according to the invention can be selected from those disclosed in WO2004/005243 and WO2004/005233, and they are in particular selected from the group consisting of:

1 - [4-chlorophenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-

2- en-1 -one, described as compound 15;

1 -[4-chlorophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2- en-1 -one, described as compound 16;

1 -[4-chlorophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1 -one as compound 17;

1 -[4-methylthiophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl] prop-2-en-1 -one, described as compound 27;

1 -[4-methylthiophenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl] prop-2-en-1 -one, described as compound 28;

1 -[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1 - one, described as compound 29; 1 -[4-hexyloxyphenyl]-3-[3,5-dimethyl-4- tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1 -one, described as compound 32;

1 -[4-hexyloxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1 -one, described as compound 33;

1 -[4-heptylphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2- en-1 -one, described as compound 38;

1 -[4-heptylphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1 -one, described as compound 39;

1 -[4-bromophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2- en-1 -one, described as compound 40;

1 -[4-bromophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1 -one, described as compound 41 .

In a further embodiment of the invention, the compounds as disclosed in WO2004/005243 and WO2004/005233 (herein referred to as compounds of General Formula (II)) that can be used and administered in combination products and/or comprised in pharmaceutical compositions according to the invention have the following General Formula (I):

in which:

X1 represents a halogen, a R1 group or a G1 -R1 group;

A represents a CH=CH group;

X2 represents a G2-R2 group;

G1 and G2, identical or different, represent an atom of oxygen or sulfur; R1 represents an alkyl or cycloalkyl group having from one to seven carbon atoms, the alkyl or cycloalkyl group being substituted or not by one or more halogen atoms;

R2 represents an alkyl group substituted by a -COOR3 group, wherein R3 represents a hydrogen atom or an alkyl group having from one to four carbon atoms.

R4 and R5 represent an alkyl group having from one to four carbon atoms.

WO2005/005369 and US20070032543 also disclose the structure and alternative process for the synthesis of compounds according to General Formula (I) as well as to General Formula (II), and they are in particular selected from the group consisting of: 1 -[4-trifluoromethylphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxy phenyl]prop-2-en-1 -one (compound 57 of US20070032543)

1 -[4-trifluoromethylphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en- 1 -one (compound 58 of US20070032543)

1 -[4-trifluoromethyloxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxy phenyl]prop-2-en-1 -one (compound 61 of US20070032543)

1 -[4-trifluoromethyloxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2- en-1 -one (compound 62 of US20070032543).

Additional examples of compounds to be used and administered and that can be comprised in compositions according to the invention can be selected from those disclosed in WO2005/073184.

In a further embodiment of the invention, the compounds as disclosed in

WO2005/073184 (herein referred to as compounds of General Formula (III)) that can be used and administered in combination products and/or comprised in pharmaceutical compositions according to the invention have the following General Formula (I):

in which:

X1 represents a G1 -R1 group;

A represents a CH=CH group;

X2 represents a G2-R2 group;

G1 and G2 represent an atom of oxygen;

R1 represents a cycloalkyl, an aryl or an alkyl group that is substituted or not by one or more alkylthio, cycloalkyl, cycloalkylthio, or heterocycloalkyl groups;

R2 represents an alkyl group substituted by at least a -COOR3 group, wherein R3 represents a hydrogen atom or an alkyl group having from one to four carbon atoms;

R4 and R5 represent an alkyl group having from one to four carbon atoms.

Additional examples of compounds to be used and administered in combination products and/or comprised in combination products or compositions according to the invention can be selected from those disclosed in WO2004/005243, WO2004/005233, WO2005/005369, US20070032543 or WO2005/073184, and reduced in the form of the corresponding substituted 1 ,3-diphenylpropanones.

Accordingly, compounds that can be used and administered in combination products and/or comprised in pharmaceutical compositions according to the invention can be selected from those disclosed in WO2007/147879, and they are in particular selected from the group consisting of:

2-[2,6-dimethyl-4-[3-[4-(methylthio)phenyl]-3-oxo-propyl]phenoxy]-2-methylpropanoic acid, described as compound 1 ;

2-[2,6-dimethyl-4-[3-[4-(methoxy)phenyl]-3-oxo-propyl]phenoxy]-2-methylpropanoic acid, described as compound 6; 2-[2,6-dimethyl-4-[3-[4-(methylthio)phenyl]-3-oxo-propyl]phenoxy]ethanoic acid, described as compound 7;

2-[2,6-dimethyl-4-[3-[4-(propyloxy)phenyl]-3-oxo-propyl]phenoxy]-2-methylpropanoic acid, described as compound 8;

2-[2,6-dimethyl-4-[3-[4-(methylthio)phenyl]-3-oxo-propyl]phenoxy]-2-methyl propanoic acid isopropyl ester, described as compound 13.

In a further embodiment of the invention, the compounds as disclosed in WO2007/147879 (herein referred as compounds of General Formula (IV)) that can be used and administered in combination products and/or comprised in pharmaceutical compositions according to the invention have the following General Formula (I):

in which:

X1 represents a R1 or a G1 -R1 group;

A represents a CH2-CH2 group;

X2 represents a G2-R2 group;

G1 represents an atom of oxygen or sulfur and G2 represents an atom of oxygen; R1 represents an alkyl group or cycloalkyl group having from one to seven carbon atoms;

R2 represents an alkyl group substituted by at least a -COOR3 group, wherein R3 represents a hydrogen atom or an alkyl group having from one to four carbon atoms; R4 and R5 represent an alkyl group having from one to four carbon atoms.

Similarly to WO2007/147879, WO2007/147880 discloses compounds to be used and administered in combination products and/or comprised in pharmaceutical compositions according to the invention that correspond to reduced, substituted 1 ,3- diphenylpropanones derivatives of compounds that were disclosed in WO2004/005243, WO2004/005233, WO2005/005369, US20070032543 or WO2005/073184, and they are in particular selected from the group consisting of:

2-[2,6-dimethyl-4-[3-[4-(trifluoromethyloxy)phenyl]-3-oxo-propyl]phenoxy]-2-methyl- propanoic acid, described as compound 1 ;

2-[2,6-dimethyl-4-[3-[4-(trifluoromethylthio)phenyl]-3-oxo-propyl]phenoxy]-2-methyl propanoic acid, described as compound 2;

2-[2,6-dimethyl-4-[3-[4-bromophenyl]-3-oxo-propyl]phenoxy]-2-methylpropanoic acid, described as compound 3;

2-[2,6-dimethyl-4-[3-[4-(trifluoromethyl)phenyl]-3-oxo-propyl]phenoxy]-2-methylpropanoic acid, described as compound 4;

2-[2,6-dimethyl-4-[3-[4-(3,3,3-trifluoropropyloxy)phenyl]-3-oxo-propyl]phenoxy]-2-methyl propanoic acid, described as compound 1 1 ;

2-(2,6-dimethyl-4-(3-oxo-3-(4-(2,2,2-trifluoroethoxy)phenyl)propyl)phenoxy)-2- methylpropanoic acid, described as compound 12;

2-(2,6-dimethyl-4-(3-oxo-3-(4-(2,2,2-trifluoroethylthio)phenyl)propyl)phenoxy)-2-methyl propanoic acid, described as compound 13;

2-(2,6-dimethyl-4-(3-oxo-3-(4-(trifluoromethoxy)phenyl)propyl)phenoxy)propanoic acid, described as compound 29;

4-(2,6-dimethyl-4-(3-oxo-3-(4-(trifluoromethoxy)phenyl)propyl)phenoxy)-2,2-dimethyl butanoic acid, described as compound 34;

2-(2,6-dimethyl-4-(3-oxo-3-(4-(trifluoromethoxy)phenyl)propyl)phenoxy)-2-methyl propanoic acid tertiobutyl ester, described as compound 35;

2-(2,6-dimethyl-4-(3-oxo-3-(4-(trifluoromethoxy)phenyl)propyl)phenoxy)-2-methyl propanoic isopropyl ester, described as compound 36;

2,2-difluoro-2-(2,6-dimethyl-4-(3-oxo-3-(4-(trifluoromethoxy)phenyl)propyl)phenoxy)acetic acid, described as compound 37. In a further embodiment of the invention, the compounds as disclosed in WO2007/147880 (herein referred to as compounds of General Formula (V)) that can be used and administered in combination products and/or comprised in pharmaceutical compositions according to the invention have the following General Formula (I):

in which:

X1 represents a halogen atom or a R1 or G1 -R1 group;

A represents a CH2-CH2 group;

X2 represents a G2-R2 group;

G1 represents an atom of oxygen or sulphur and G2 represents an atom of oxygen; R1 represents an alkyl or cycloalkyl group that is substituted or not by one or more halogen atoms;

R2 represents an alkyl group substituted or not by one or more halogen atoms and substituted by at least a -COOR3 group, wherein R3 represents a hydrogen atom or an alkyl group having from one to four carbon atoms.

R4 and R5 represent an alkyl group having from one to four carbon atoms.

The compounds that are preferably used, administered in combination products and/or comprised in pharmaceutical compositions according to the invention are those defined according to General Formula (II) or General Formula (V). Most preferably, the compounds are selected from the ones those listed below:

1 -[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1 - one (compound 29 of WO2004/005233 described in the Examples as Cpd A)

1 -[4-methylthiophenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyph prop-2-en-1 -one (compound 28 of WO2004/005233)

1 -[4-methylthiophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl] prop-2-en-1 -one (compound 27 of WO2004/005233)

1 -[4-trifluoromethylphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxy phenyl]prop-2-en-1 -one (compound 57 of US20070032543)

1 -[4-trifluoromethylphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en- 1 -one (compound 58 of US20070032543 described in the Examples as Cpd B)

1 -[4-trifluoromethyloxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimeth phenyl]prop-2-en-1 -one (compound 61 of US20070032543)

1 -[4-trifluoromethyloxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2- en-1 -one (compound 62 of US20070032543)

2-[2,6-dimethyl-4-[3-[4-(trifluoromethyloxy)phenyl]-3-oxo-propyl]phenoxy]-2- methylpropanoic acid (compound 1 of WO 2007147880 described in the Examples as Cpd C)

2-[2,6-dimethyl-4-[3-[4-(trifluoromethylthio)phenyl]-3-oxo-propyl]phenoxy]-2- methylpropanoic acid (compound 2 of WO 2007147880)

2-(2,6-dimethyl-4-(3-oxo-3-(4-(trifluoromethoxy)phenyl)propyl)phenoxy)-2-methyl propanoic acid tertiobutyl ester (compound 35 of WO 2007147880).

As aforementioned, the present invention provides uses of compounds of General Formula (I) in combination with a DPP-4 Inhibitor and related pharmaceutical compositions comprising the same. The compound and the DPP-4 Inhibitor may or may not be in the form of their pharmaceutically acceptable salts and are used in combination in a therapeutically effective amount for treating conditions mediated by GLP-1. The compound of General Formula (I) exerts its action by potentiating the DPP-4 Inhibitor effect on endogenous GLP-1 level. Any compound that is defined according to General Formula (II), General Formula (III), General Formula (IV) or General Formula (V) can be used according to the present invention for treating conditions mediated by GLP-1 , in particular in the form of a pharmaceutical composition that comprises said compound in combination of DPP-4 Inhibitor.

The invention also provides a method for the treatment of conditions mediated by GLP-1 comprising the administration in a therapeutically effective amount to a subject in need thereof of the combination of a DPP-4 Inhibitor and of a compound of General formula (I) in which:

X1 represents a halogen, a R1 or G1 -R1 group;

A represents a CH=CH or a CH2-CH2 group;

X2 represents a G2-R2 group;

G1 and G2, identical or different, represent an atom of oxygen or sulfur;

R1 represents a hydrogen atom, an unsubstituted alkyl group, a cycloalkyl group, an aryl group or an alkyl group that is substituted by one or more halogen atoms, alkoxy, alkylthio, cycloalkyl, cycloalkylthio, or heterocyclic groups; R2 represents an alkyl group substituted by at least a -COOR3 group, wherein R3 represents a hydrogen atom, or an alkyl group that is substituted or not by one or more halogen atoms, cycloalkyl groups, or heterocyclic groups;

R4 and R5, identical or different, representing an alkyl group that is substituted or not by one or more halogen atoms, cycloalkyl groups, or heterocyclic groups;

or the administration of a pharmaceutical composition comprising said combination.

Compounds wherein X1 , X2, A, G1 , G2, R1 , R2, R3, R4 and R5 are defined according to General Formula (II), General Formula (III), General Formula (IV) or General Formula (V) and in particular according to General Formula (II) or General Formula (V) can also be used to carry out the method of the treatment of the invention.

The compound of General Formula (I) and the DPP-4 Inhibitor may contain one or several asymmetrical centers. When an enantiomerically pure (or enriched) preparation of any of these compounds is desired, it can be obtained either by purification of the final product or chiral intermediates, or by asymmetrical synthesis following the typical methods known by one of ordinary skill in the art (for example, by using chiral reactants and catalysts). Some of these compounds can have different stable tautomeric forms. This invention includes the use of stereoisomers (diastereoisomers, enantiomers), pure or mixed, as well as racemic mixtures and geometrical isomers of compounds of General Formula (I) and of the DPP-4 Inhibitor for the manufacture of pharmaceutical compositions for treating a condition mediated by GLP-1 .

The combinations of a compound of General Formula (I) and of a DPP-4 Inhibitor can be formulated as "pharmaceutically acceptable" salts, being slightly- or non-toxic salts obtained from organic or inorganic bases or acids of such compounds. Alternatively, the combinations of a compound of General Formula (I) and of a DPP-4 Inhibitor can be formulated as "pharmaceutically acceptable" hydrates or polymorphs of such compounds. These salts, hydrates, and polymorphs can be obtained during the final purification step of the compound or, in the case of salts, by incorporating the salt into the previously purified compound.

The present invention further provides pharmaceutical compositions comprising a DPP-4 Inhibitor and at least one further compound of General Formula (I), or their pharmaceutically acceptable salts, and optionally at least one pharmaceutically acceptable carrier or diluent, in particular for use in a method of treatment of a condition mediated by GLP-1 . The pharmaceutical compositions comprising a compound of General Formula (I) and of a DPP-4 Inhibitor may comprise one or several excipients or vehicles, acceptable within a pharmaceutical context (e.g., for liquid formulations, saline solutions, physiological solutions, isotonic solutions, that are compatible with pharmaceutical usage).

A further object of the invention are methods of preparing such pharmaceutical compositions admixing a compound of General Formula (I) and a DPP-4 Inhibitor, together with at least one pharmaceutically acceptable carrier, vehicle, or diluent. These methods are well known in the art, and involve, for example, conventional mixing, dissolving, granulation, dragee-making, levigating, emulsifying, encapsulating, entrapping, lyophilizing processes or spray drying (Remington: The Science and Practice of Pharmacy, 20th Edition, 2000, Lippincott Williams & Wilkins; Handbook of Pharmaceutical Excipients, 4th Edition, 2003, Pharmaceutical Press).

The term "pharmaceutically acceptable" refers to those properties and/or substances that are acceptable to the patient from a pharmacological/toxicological point of view and to the manufacturing pharmaceutical chemist from a physical/chemical point of view regarding composition, formulation, stability, patient acceptance and bioavailability.

The term "carrier", "vehicle", or "excipient" refers to any substance, not itself a therapeutic agent, that is added to a pharmaceutical composition to be used as a carrier, vehicle, and/or diluent for the delivery of a therapeutic agent to a subject in order to improve its handling or storage properties or to permit or facilitate formation of a dosage unit of the composition into a discrete article. The pharmaceutical compositions of the invention, either individually or in combination, can comprise one or several agents or vehicles chosen among dispersants, solubilisers, stabilisers, preservatives, etc. Agents or vehicles useful for these formulations (liquid and/or injectable and/or solid) are particularly methylcellulose, hydroxymethylcellulose, polysorbate 80, mannitol, gelatin, lactose, vegetable oils, liposomes, etc. Acceptable excipients can be chosen among disintegrants, binding agents, adhesives, wetting agents, lubricants, glidants, flavors, dyes, fragrances, stearic acid, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, magnesium carbonate, talc, gelatin, lactose, sucrose, starches, polymers, such as polyvinyl alcohol and polytheylene glycols, and other pharmaceutically acceptable materials added to improve taste, odor, or appearance of the composition.

The compounds can be made up in solid or liquid form, such as tablets, capsules, powders, syrups, elixirs and the like, aerosols, sterile solutions, suspensions or emulsions, and the like. The composition may be presented in a solid preformulation composition wherein the active ingredients are dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective dosage forms such as tablets, pills and capsules. Additionally, the combined compositions may be delivered using sustained-release formulations.

The compositions can be formulated in the form of injectable suspensions, gels, oils, pills, suppositories, powders, gel caps, capsules, aerosols, etc., eventually by means of galenic forms or devices assuring a prolonged and/or slow release. For this kind of formulation, agents such as cellulose, carbonates or starches can advantageously be used. The compositions of the present invention can also be formulated in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of lipids, including but not limited to amphipathic lipids such as phosphatidylcholines, sphingomyelins, phosphatidylethanolamines, phophatidylcholines, cardiolipins, phosphatidylserines, phosphatidylglycerols, phosphatidic acids, phosphatidylinositols, diacyl trimethylammonium propanes, diacyl dimethylammonium propanes, and stearylamine, neutral lipids such as triglycerides, and combinations thereof.

The pharmaceutical combination of the invention can be administered in a systematic or parenteral way, by using oral, nasal, rectal, transmucosal, transdermal, intestinal, intramuscular, intravenously, subcutaneous, intraarterial, intraperitoneal, intrapulmonary, or intraocular route, by using methods known in the art.

Oral administration is the preferential route of administration for pharmaceutical compositions comprising a compound of General Formula (I) and the DPP-4 Inhibitor for the treatment of a condition mediated by GLP-1 . Formulations for oral administration may be in the form of aqueous solutions and suspensions, in addition to solid tablets and capsule formulations. The aqueous solutions and suspensions may be prepared from sterile powders or granules. The compounds may be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, and/or various buffers.

The tablets or pills of the composition can be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permits the inner component to pass intact into the duodenum or to be delayed in release. A variety of material can be used for such enteric layers or coatings, such materials including a number of polymeric acids with such as shellac and cellulose acetate.

The liquid forms in which the pharmaceutical compositions can be incorporated for oral administration or by injection include, aqueous solutions, suitably flavoured syrups, aqueous or oil suspensions, and flavoured emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical vehicles. Suitable dispersing or suspending agents for aqueous suspensions, include synthetic and natural gums such as tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinyl-pyrrolidone or gelatin. The liquid forms in suitably flavored suspending or dispersing agents may also include the synthetic and natural gums, for example, tragacanth, acacia, methyl-cellulose and the like. For parenteral administration, sterile suspensions and solutions are desired.

A pharmaceutical composition as disclosed herein is understood to be useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, that is, the active ingredients are contained in an amount to achieve their intended purpose. At this scope, the combination of a compound of General Formula (I) and of a DPP-4 Inhibitor should be administered in an effective quantity by using a pharmaceutical composition as above-defined. Administration can be performed daily or even several times per day, if necessary, and in an amount that can be optimal or suboptimal, if they are compared with dosages that are normally used for such compounds.

The term "an effective quantity" refers to an amount of the compound sufficient to produce the desired therapeutic result, in particular the compounds of General Formula (I) and the DPP-4 Inhibitor are administered in amounts that are sufficient to increase a blood GLP-1 level in a subject. In particular, the compound of General Formula (I) and the DPP- 4 Inhibitor are formulated in a dosage form that should be sufficient to lower a blood glucose level in a subject, improve glucose control, increase the amount of insulin, and/or increase the sensitivity of cells to insulin following the simultaneous, separate or sequential administration of the two compounds.

The combination product according to the invention for use in a method for the treatment of a condition mediated by GLP-1 (Glucagon-Like Peptide 1 ) is for a simultaneous use of the two compounds, or said two compounds are comprised in two separate dosage forms. More particularly, the combined administration of compound of General Formula (I) and of DPP-4 Inhibitor can be performed by simultaneously administering the two compounds using a single pharmaceutical composition (in the form of a pill, a tablet, etc.). Alternatively, the combination product comprising the compound of General Formula (I) and the DPP-4 Inhibitor can be provided in two distinct dosage forms for simultaneous (for example, using two different pills), separate or sequential (at different times of the day or in different days) use and administration. Accordingly, the combined administration of compound of General Formula (I) and the DPP-4 Inhibitor can be performed as a "kit-of-parts".

The present invention consequently relates to a "kit-of-parts" for separate, sequential or simultaneous use in a method for the treatment of a condition mediated by GLP-1 (Glucagon-Like Peptide 1 ) comprising at least two dosage forms of components:

(a) a pharmaceutical formulation including a Dipeptidyl Peptidase 4 (DPP-4) Inhibitor and

(b) a pharmaceutical formulation including a compound of General Formula (I).

These formulations can be prepared such as to be suitable for separate (for example, at different times of the day or in different days) or simultaneous (for example, using two different pills, tablets, etc.) use in a method for the treatment of a condition mediated by GLP-1 . Preferably, in the case of a separate administration, the formulations are suitable for sequential administration; for example, the pharmaceutical formulation (b) is administered before the pharmaceutical formulation (a).

These two distinct dosage forms in the form of the kit-of-parts according to the invention can be provided in a commercial package comprising a compound of General Formula (I) and a DPP-4 Inhibitor in the form of pharmaceutical compositions comprising the same, together with instructions, for simultaneous, separate or sequential use thereof in the treatment of a condition mediated by GLP-1 , in particular a metabolic disorder.

The term "combined administration" includes co-administration wherein: 1 ) the two or more agents are administered to a subject at substantially similar times; and 2) the two or more agents are administered to a subject at different times, at independent intervals which may or may not overlap or coincide. Optimal dosages to be administered may be readily determined by those skilled in the art, and will vary with the particular compound used, the strength of the preparation, the mode of administration, and the severity of the condition to be treated. In addition, factors associated with the particular patient being treated, including patient age, weight, diet and time of administration, will result in the need to adjust dosages and interval, in particular for providing a synergistic effect in lowering blood glucose level in a subject. The frequency and/or dose relative to the simultaneous or separate administrations can be adapted by one of ordinary skill in the art, in function of the patient, the pathology, the form of administration, etc. For instance, the combination of a compound of General Formula (I) and of the DPP-4 Inhibitor should be provided in the dosage that allows their administration of each compound in the amount 0.01 mg/day to 1000 mg/day, preferably from 0.1 mg/day to 10 mg/day.

The pharmaceutical compositions according to the invention can be advantageously formulated and administered in combination with other therapeutic agents, currently available in the market or in development, in particular for the treatment of metabolic disorders, such as metformin, insulin, thiazolidinediones, glitazones, statins, inhibitors of cholesterol and/or other lipid lowering drugs.

All references cited herein are fully incorporated by reference in their entirety. Having now fully described the invention, it will be understood by those of ordinary skill in the art that the invention may be practiced within a wide and equivalent range of conditions, parameters and the like, without affecting the spirit or scope of the invention or any embodiment thereof. Several other advantages of the invention will rise in the reading of the following examples; they should be considered as illustrative data and not as limitative ones.

EXAMPLES Abbreviations used in the Examples: - Cpd A: compound 29 of WO2004/005233

- Cpd B: compound 58 of US2007/0032543

- Cpd C: compound 1 of WO2007/147880

- DMSO: dimethyl sulfoxide

- DPP-4: dipeptidyl peptidase-4

- ELISA: enzyme-linked immunosorbent assay

- GLP-1 : glucagon-like peptide 1

- oGTT: oral glucose tolerance test

- Vs.: versus

Example 1 : In vivo effects on GLP-1 secretion following the combined administration of a DPP-4 Inhibitor with a compound of General Formula (I)

Materials & Methods

Animal model

Male Sprague Dawley rats (Charles River, Saint-Germain-sur-l'Arbresle, France) were 8 weeks old at the beginning of the experiment. The rats were given a standard diet (Safe, Augy, France). The animals were kept in a 12 hours/12 hours light/dark cycle at a constant temperature of 20 +/- 3°C, with free access to water and food. The body weight was recorded on a daily basis. After one week acclimation, the rats were separated in 3 groups of 8 animals each, selected so that the body weight and the overnight fasting glycemia were similar within a given group at the beginning of the experiment.

The compound to be tested was suspended in an aqueous 1 % carboxymethylcellulose solution (Sigma Aldrich, C4888) containing 0.1 % Tween 80 (Sigma Aldrich, P8074) and administered by intragastric gavage, once a day for 13 days. On day 14, after 24 hours of food fasting and 1 .5 hours of water fasting, Sitagliptin (DPP-4 Inhibitor; 25 mg/kg; 5ml/kg) was orally administrated to rats and, 30 minutes later, the animals were challenged with an oral glucose load (3 g/kg; 10ml/kg). Control animals were treated with vehicle only (Carboxymethlycellulose 1 % + Tween-80 0.1 %).

Blood samples were harvested for GLP-1 determination by sub-lingual bleed 0 and 5 minutes after the glucose load. Plasma was obtained by centrifugation at 4,000 rpm (4°C, 15 minutes) and stored at -80°C until the dosage.

GLP-1 quantification

The plasma GLP-1 concentration was determined by an enzyme-linked immunosorbent assay (ELISA) technique that measured the biologically active GLP-1 forms GLP-1 (7-36) and GLP-1 (7-37) amide (Millipore, Molsheim, France; Cat. No. EGLP- 35K) according to the manufacturer's recommendations.

Briefly, 100 microliters of 1 :2 diluted plasma samples were distributed in the ELISA plate and kept during 24 hours at 4°C for binding to the immobilized anti-GLP-1 monoclonal antibody. The next day, several washes were performed and 200 microliters of anti-GLP-1 -alkaline phosphatase Conjugate were dispatched in each well and the plate was incubated 2 hours at room temperature. After several washes, the substrate (4- Methylumbelliferyl Phosphate) was added in each well and the plate was kept at least 20 minutes at room temperature in the dark. At the end of the reaction, 50 microliters of Stop Solution were added to each well and incubated 5 minutes at room temperature in the dark before fluorescence reading using the Genius Pro® (Tecan, Lyon, France) with an excitation/emission wavelength of 355 nm/460 nm.

Statistic analysis of plasma GLP-1 concentration was performed using unpaired Student's t-test with three p values that defines statistical relevance (^* means p<0.05; ^** means p<0.01 ).

Results & Conclusions:

The activity of a compound of General FormulaJI) has been tested in association with DPP-4 Inhibitors in Sprague Dawley rats, an animal model commonly used for assessing the effects of compounds and/or food on plasma GLP-1 concentration and secretion ( Burkey B et al., 2005; Yoder S et al., 2009).

The objective of this study was to evaluate, in vivo, the effect on GLP-1 secretion that was induced by the oral chronic treatment of rats with Cpd A (a compound of General Formula (I) and more precisely of General Formula (II)) during 13 days, followed by an acute treatment with a DPP-4 Inhibitor at day 14, 30 minutes before receiving the oral glucose load in an oGTT. The effects of the chronic oral treatment of rats with Cpd A (30 mg/kg/day) or with the PPARgamma activator Rosiglitazone (10 mg/kg/day) were compared to those of the control group that received vehicle only. The plasma GLP-1 levels were measured to determine the fold induction in GLP-1 plasma secretion by comparing basal level (before the glucose load) and the GLP-1 plasma level in the same animals 5 minutes after the glucose load. Table 1 shows that only Cpd A induced the secretion of GLP-1 in the plasma of Sprague Dawley rats 5 minutes after the glucose load in a statistically significant manner.

TABLE 1

Thus, compounds of General Formula (I) (and in particular a compound of General Formula (II)) that have been characterized as activators of PPAR nuclear receptors, as shown in WO 2004/005233, have in vivo effects on the potentiation of the activities of DPP-4 Inhibitors unexpectedly superior to those observed by using a marketed antidiabetic compound such as Rosiglitazone (Roy S et al., 2007) that targets the same family of biological targets, i.e. PPAR nuclear receptors. The chronic administration of a compound of General Formula (I) can consequently provide a major benefit when associated with the administration with a DPP-IV inhibitor for treating a condition mediated by GLP-1 and in particular metabolic disorders such as type 2 diabetes.

This experimental protocol can be adapted for testing specific pharmaceutical combinations according to the invention by using different compounds of General Formula (I) (and in particular of General Formula (II), (III), (IV), or (V)), DPP-4 Inhibitors, regimens, dosages, and/or formulations of these compounds. The most preferred features of the invention about the potentiating effects on DPP-4 activities, can be then determined by using assays for measuring plasma GLP-1 levels following an oral glucose or meal load, similarly to the experiments described above, before performing other tests about the improved GLP-1 regulatory effects on glucose homeostasis and on extra-pancreatic activities in treated animals and/or human subjects.

Example 2: In vivo models for testing the combined administration of DPP-4 Inhibitors and compounds of General Formula (I) for the treatment of type 2 diabetes

Materials & Methods

Animal model

Male insulin-resistant ob/ob mice (Charles River, Saint-Germain sur I'Arbresle, France) were 8 weeks old at the beginning of the experiment. Mice were kept on a 12 hours/12 hours light/dark cycle at a constant temperature of 20 ± 3°C, with free access to water and food. Animals were fed a standard chow diet (R03, SAFE, Augy, France). After one week acclimation, mice were separated in groups of 8 animals selected so that the distribution of their body weight and the 6 hours fasting glycemia determined before the experiment were similar with a given group at the beginning of the experiment.

Compounds to be tested were suspended in an aqueous 1 % carboxymethylcellulose solution (Sigma Aldrich, C4888) containing 0.1 % Tween-80 (Sigma Aldrich, P8074) and administered by intragastric gavage, once a day for 21 days. Control animals were treated with vehicle only (Carboxymethlycellulose 1 % + Tween-80 0.1 %).

GLP-1 secretion assay

On day 21 , a GLP-1 secretion assay was carried out. Briefly, after 24 hours of food fasting and 1.5 hours of water fasting, Sitagliptin (DPP-4 Inhibitor; 25 mg/kg, 5 ml/kg) was orally administrated to the animals and, 45 minutes later, the animals were challenged with an oral glucose load (3 g/kg, 10 ml/kg). Blood samples were harvested for GLP-1 determination by retro-orbital puncture 0 and 15 minutes after the glucose load. Plasma was obtained by centrifugation at 4,000 rpm (4°C, 15 minutes) and stored at -80°C until the dosage. The measurement of GLP-1 concentration in the plasma samples and the statistical analysis of these data were performed as described in Example 1.

Results & Conclusions:

The anti-diabetic effect of GLP-1 is usually explored in a pathological in vivo model that reflects the Diabetes mellitus type 2 symptoms. A number of animal models of Type 2 diabetes are available that encompass obesity and insulin resistance, both features of Type 2 diabetes in humans (Rees D and Alcolado J, 2005). The Zucker Diabetic Fatty (ZDF) rats is one of them and is commonly used to investigate the antidiabetic effects of DPP-4 Inhibitors that are administered as a monotherapy or in association with other drugs (Brand C et al., 2009; Pei Z et al., 2006; Sudre B et al., 2002).

The global antidiabetic effect and the increase in GLP-1 secretion, as demonstrated above by the combined administration of the invention, can be also assessed in ZDF rats that are chronically treated with a DPP-4 Inhibitor and a compound of General Formula (I) during 7-28 days. The ZDF rats can be given a standard diet and kept in a 12 hours/12 hours light/dark cycle at a constant temperature of 20 +/- 3°C. Groups of animals (one for each type of combined administration and consisting of 5-12 rats) can be established so that features such as body weight, glycemia, and other relevant plasma indexes are similar for all animals within a given group at the beginning of the experiment.

The combined and control treatments are administrated every day by oral intragastric gavage. For instance, one group receives Sitagliptine only (25 mg/kg/day), one group receives a compound of the General Formula (I), one group receives the combination of Sitagliptine (25 mg/kg/day) and Cpd of the General Formula (I) and the control group receives vehicle only. After 14 or more days of treatment and a 24 hours period of food fasting, blood samples are harvested by sub-lingual bleed for measuring glycated haemoglobin (HblAc), glucose, insulin and GLP-1 levels in plasma using standard protocols. Such parameters are compared among the different groups in order to compare different ratio in the amount of administered compounds, alternative compounds of General Formula (I), and/or alternative DPP-4 Inhibitors).

Another relevant animal model is the insulin-resistant ob/ob mice which is commonly used for assessing the effects of anti-diabetic effect of GLP-1 and which has been used for testing the combination of a DPP-4 Inhibitor with pioglitazone (Moritoh Y et al., 2009).

The objective of this study was to evaluate, in vivo, the effect on GLP-1 secretion that was induced by the oral chronic treatment of insulin resistant mice during 21 days with specific compounds of General Formula (I) followed by an acute treatment with a DPP-4 Inhibitor at day 21 , 30 minutes before receiving the oral glucose load in an oGTT. The effects of the chronic oral treatment of the ob/ob mice during 21 days with Cpd A (30 mg/kg/day), Cpd B (30 mg/kg/day), or the Cpd C (30 mg/kg/day) were compared to those of the control group that received vehicle only.

The plasma GLP-1 levels were measured to determine the fold induction in GLP-1 plasma secretion by comparing basal level (before the glucose load) and the GLP-1 plasma level in the same animals 15 minutes after the glucose load. Secretion levels were then normalized to the control group. Table 2 shows that the Cpds A, B and C, when compared to controls, induced in a statistically significant manner a higher secretion of GLP-1 in the plasma of insulin-resistant ob/ob mice, 15 minutes after the glucose load.

TABLE 2

These data clearly indicate that the conclusions made on a compound of General Formula (I) in Example 1 on the major potential benefit of the administration in combination with a DPP-IV inhibitor, apply also to other compounds belonging to General Formula (I), and in particular of General Formula (II) (such as Cpd A and Cpd B) and (V) (such as Cpd C). These results further confirm the positive effects of a combined administration according to the invention on the treatment of type 2 diabetes.

Example 3: In vitro/ex vivo models for evaluating the induction of GLP-1 secretion by compounds of General Formula (I)

Materials & Methods

Cell culture and treatment with compounds of the General Formula (I)

GLUTag cells were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 1000 mg/L glucose (Invitrogen, Cat. No. 21885-025) and supplemented with 10% fetal bovine serum, 2mM glutamine and 1 % antibiotics (Penicillin/ Streptomycin) until they reach 80% confluence. Then, GluTag cells were washed with Krebs buffer and incubated for 30 minutes with Krebs buffer supplemented with 100 μΜ of DPP-4 Inhibitor (Diprotin A, Sigma Aldrich). Afterwards, cells were treated during 30 minutes in Krebs buffer supplemented with 100 μΜ of DPP-4 Inhibitor with 1 % DMSO alone (negative control), or with the Cpd A ( at 10 mM suspended in 1 % DMSO). At the end of the experiment, cell culture medium was harvested for GLP-1 quantification. The measurement of GLP-1 concentration in cell culture medium and the statistical analysis of these data were performed as described in Example 1 for plasma samples.

Results & Conclusions:

Intestinal L-cells secrete GLP-1 in response to ingestion of nutrients, especially long-chain fatty acids. On this basis, some in vitro models have been established for screening compounds that can have a positive effect on GLP-1 secretion (Reimann F et al. 2006). In particular, GLUTag L-cell line has been developed from intestinal endocrine tumors and secretes GLP-1 following specific stimulation (Brubaker P et al., 1998), for instance by using reference compound known to induce GLP-1 in this cell line such as a GPR1 19 agonist (Lauffer L et al., 2009) or a neurotransmitter (Garneiro A et al., 2005)

When compared with the control cells, Cpd A increased the GLP-1 secretion in 24hour-treated GLUTag cells in a statistically significant manner (1.6-fold; p<0.01 ). The data on the in vitro response of GLUTag cells to the exposure to Cpd A confirm that compounds of General Formula (I) can be accordingly tested and compared at different concentrations (for example 3, 10, 30 micromolar) for establishing further criteria for selecting those to be tested and compared by using the in vivo models described in the Examples above and in the literature.

REFERENCES

Ahren B, 2009. Best Pract Res Clin Endocrinol Metab; 23: 487-98.

Bolen S et al., 2007. Ann Intern Med; 147: 386-99.

Brand C et al., 2009. Diabetes Obes Metab; 1 1 : 795-803.

Brubaker P et al., 1998. Endocrinology; 139; 4108-41 14.

Burdo J et al., 2009. Neurobiol Aging ; 30: 1910-1917.

Burkey B et al., 2005. J Pharmacol Exp Ther.; 315: 688-95.

Buteau J, 2008. Diabetes Metab ; 34 SuppI 2:S73-7.

Campbell I, 2009. Brit J Diab Vase Dis; 9: 53-63.

Campbell R, 2009a. J Am Pharm Assoc; 49 SuppI 1 : S10-15.

Campbell R, 2009b. J Am Pharm Assoc ; 49 SuppI 1 : S3-9.

Conarello S et al., 2003. Proc Natl Acad Sci U S A; 100: 6825-30. de la Monte S and Wands J, 2008. J Diab Sci Tech; 2: 1 101 -1 1 13. Drucker D, 2006. Lancet ; 368: 1696-705.

Drucker D, 2007. Diabetes Care ; 30: 1335-43.

Garneiro A et al., 2005. J Physiol ; 569: 761 -72.

Gupta R et al., 2009. Curr Drug Targets; 10: 71 -87.

Hansen K et al., 2009. Int J Clin Pract ; 63: 1 154-60.

Havale S and Pal M, 2009. Bioorg Med Chem ; 17: 1783-1802.

Inzucchi s and McGuire D, 2008. Circulation^ 17: 574-84.

Lauffer L et al., 2009. Diabetes ; 58: 1058-66.

Levien T and Baker D, 2009. Diabetes Spectrum ; 22: 92.

Moritoh Y et al., 2009. Eur J Pharmacol ; 602: 448-54.

Neumiller Jet al., 2009. J Am Pharm Assoc.; 49 SuppI 1 :S16-29.

Pei Z et al., 2006. J Med Chem ; 49 : 6439-6442.

Pfutzner A et al., 2008. Clin Lab ; 54: 485-90.

Pratley R and Salsali A, 2007. Curr Med Res Opin; 23: 919-31. Ranganath L et al., 2008. Clin Chem Lab Med; 46: 43-56.

Reimann F et al. 2006. Diabetes; 55 (Suppl. 2): S78-S85.

Rees D and Alcolado J, 2005. Diabet Med; 22: 359-370.

Rosenstock J et al., 2007. Diabetes Obes Metab; 9: 175-85.

Roy S et al., 2007. Life Sci; 81 : 72-9.

Seufert J, 2009. Fundam Clin Pharmacol; 23: 651 -67.

Sudre B et al., 2002. Diabetes; 51 , 1461 -1469.

Wajchenberg B et al, 2007. Endocr Rev; 28: 187-218.

White J, 2009. J Am Pharm Assoc; 49 Suppl 1 : S30-40.

Yoder S et al., 2009. Am J Physiol Gastrointest Liver Physiol ; 297: G299-305.

Claims

1 . A combination product comprising a Dipeptidyl Peptidase 4 (DPP-4) Inhibitor and a compound of General Formula (I)

in which:

X1 represents a halogen, a R1 or G1 -R1 group;

A represents a CH=CH or a CH2-CH2 group;

X2 represents a G2-R2 group;

G1 and G2, identical or different, represent an atom of oxygen or sulfur;

R1 represents a hydrogen atom, an unsubstituted alkyl group, a cycloalkyl group, an aryl group or an alkyl group that is substituted by one or more halogen atoms, alkoxy, alkylthio, cycloalkyl, cycloalkylthio, or heterocyclic groups;

for use in a method for the treatment of a condition mediated by Glucagon-Like

Peptide 1 .

2. The combination product of claim 1 , wherein the compound is of general formula (I)

in which:

X1 represents a halogen, a R1 , or a G1 -R1 group;

A represents a CH=CH group;

X2 represents a G2-R2 group;

G1 and G2, identical or different, represent an atom of oxygen or sulfur;

R1 represents an alkyl or cycloalkyi group having from one to seven carbon atoms, the alkyl or cycloalkyi group being substituted or not by one or more halogen atoms; R2 represents an alkyl group substituted by a -COOR3 group, wherein R3 represents a hydrogen atom or an alkyl group having from one to four carbon atoms; R4 and R5 represent an alkyl group having from one to four carbon atoms.

3. The combination product of claim 1 , wherein the compound is of general formula (I)

in which:

X1 represents a halogen atom or a R1 or G1 -R1 group;

A represents a CH2-CH2 group;

X2 represents a G2-R2 group;

G1 represents an atom of oxygen or sulfur and G2 represents an atom of oxygen; R1 represents an alkyl or cycloalkyi group that is substituted by one or more halogen atoms; R2 represents an alkyl group substituted or not by one or more halogen atoms and substituted by at least a -COOR3 group, wherein R3 represents a hydrogen atom or an alkyl group having from one to four carbon atoms;

R4 and R5 represent an alkyl group having from one to four carbon atoms.

4. The combination product of any one of the preceding claims, wherein the compound is of general formula (I) in which X1 is a G1 -R1 group.

5. The combination product of any one of the preceding claims, wherein the compound is of general formula (I) in which G2 is an oxygen atom and R2 is an alkyl group substituted by a -COOR3 group, wherein R3 represents a hydrogen atom or an unsubstituted linear or branched alkyl group having from one to four carbon atoms.

6. The combination product of any one of the preceding claims, wherein the compound is of general formula (I) in which X1 is an alkylthio group that comprises an alkyl group that is linear or branched, having from one to seven carbon atoms that is substituted or not by one or more halogen atoms.

7. The combination product of claim 1 , wherein said compound is of general formula (I) and is selected in the group consisting of 1 -[4-methylthiophenyl]-3-[3,5-dimethyl-4- carboxydimethylmethyloxyphenyl]prop-2-en-1 -one, 1 -[4-methylthiophenyl]-3-[3,5- dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl] prop-2-en-1 -one, 1 -[4- methylthiophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl] prop-2-en-1 -one, 1 -[4-trifluoromethylphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyl dimethylmethyloxyphenyl]prop-2-en-1 -one, 1 -[4-trifluoromethylphenyl]-3-[3,5- dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1 -one, 1 -[4-trifluoromethyl oxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxy phenyl] prop-2- en-1 -one, 1 -[4-trifluoromethyloxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyl oxyphenyl]prop-2-en-1 -one, 2-[2,6-dimethyl-4-[3-[4-(trifluoromethyloxy)phenyl]-3- oxo-propyl]phenoxy]-2-methylpropanoic acid, 2-[2,6-dimethyl-4-[3-[4-

(trifluoromethylthio)phenyl]-3-oxo-propyl]phenoxy]-2-methylpropanoic acid and 2- [2,6-dimethyl-4-[3-oxo-3-[4-(trifluoromethyloxy)phenyl]propyl]phenoxy]-2- methylpropanoic acid tert-butyl ester.

8. The combination product of any one of the preceding claims, wherein the DPP-4 Inhibitor is selected in the group consisting of Sitagliptin, Vildagliptin, Saxagliptin, Alogliptin and the pharmaceutically acceptable salts thereof.

9. A pharmaceutical composition comprising the combination product as defined in any one of the preceding claims and a pharmaceutically acceptable carrier for use in a method for the treatment of a condition mediated by GLP-1.

10. The pharmaceutical composition of claim 9, wherein said composition is formulated in the form of injectable suspensions, gels, oils, pills, suppositories, powders, gel caps, capsules, aerosols or means of galenic forms or devices assuring a prolonged and/or slow release.

1 1 . A kit-of-parts for separate, sequential or simultaneous use in a method for the treatment of a condition mediated by GLP-1 (Glucagon-Like Peptide 1 ) comprising at least two dosage forms of components:

(a) a pharmaceutical formulation including a Dipeptidyl Peptidase 4 (DPP-4) Inhibitor;

(b) a pharmaceutical formulation including a compound of General Formula (I)

in which:

X1 represents a halogen, a R1 or G1 -R1 group;

A represents a CH=CH or a CH2-CH2 group;

X2 represents a G2-R2 group;

G1 and G2, identical or different, represent an atom of oxygen or sulfur;

The kit-of-parts of claim 1 1 for sequential use in a method for the treatment of a condition mediated by GLP-1 wherein the pharmaceutical formulation (b) is administered before the pharmaceutical formulation (a).

The combination product of any one of the claims 1 to 8 or the pharmaceutical composition of claim 9 or 10 or the kit-of-parts of claim 1 1 or 12 for use in a method for the treatment of a condition mediated by GLP-1 selected in the group consisting of metabolic disorders, disorders related to pancreatic beta-cells number and/or activity, and neurodegenerative disorders. The combination product of any one of the claims 1 to 8, the pharmaceutical composition of claim 9 or 10 or the kit-of-parts of claim 1 1 or 12 for use in a method for the treatment of a metabolic disorder, wherein the metabolic disorder is selected in the group consisting of diabetes, obesity, and a disorder related to diabetes.