WO2007128801A1

WO2007128801A1 - Combination of organic compounds

Info

Publication number: WO2007128801A1
Application number: PCT/EP2007/054379
Authority: WO
Inventors: Nigel Scott Cook; Kurt Chum Graves
Original assignee: Novartis Ag
Priority date: 2006-05-08
Filing date: 2007-05-04
Publication date: 2007-11-15

Abstract

This invention relates to a pharmaceutical combination comprising a DPP-IV inhibitor, and i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or ii) omega-3 oils, in particular for the prevention, delay of progression or treatment of diseases and disorders that may be inhibited by DPP IV inhibition.

Description

Combination of Organic Compounds

This invention relates to a pharmaceutical combination comprising a DPP-IV inhibitor, and i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or ii) omega-3 oils, in particular for the prevention, delay of progression or treatment of diseases and disorders that may be inhibited by DPP IV inhibition.

The hypotriglyceridemic effects of omega-3 oils from fish oils are well established. Amounts both above and below about 1 gram per day of omega-3 oils from fish oil have been shown to decrease serum triglyceride concentrations by about 25 % to about 40 %, decrease VLDL blood plasma levels, and to increase both LDI and HDL plasma levels (See e.g., Harris, William S. Clin. Cardiol. 22, (Suppl. II), II-40-II- ] 43 (1999)). A dose-response relationship exists between omega-3 oil intake and triglyceride lowering. Postprandial triglyceridemia is especially sensitive to chronic omega-3 oil consumption. Kris-Etherton, et al., Circulation. 2002;106:2747.

There are three distinct components of omega 3 fatty acids, which have been shown to be beneficial at various life stages and with several different health conditions. These are DHA (docosahexaenoic acid) and EPA (eicosapentaenoic acid), as well as ALA (alpha-linolenic acid), which has to be converted to EPA and DHA in the body in order for it to be beneficial. ALA can be found in flaxseed oil and to a lesser extent, canola, soy, perilla and walnut oils.

It has now been found that a combination comprising a DPP-IV inhibitor e.g. as defined below and, at least one compound selected from the group consisting of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, e.g. as defined below, has a beneficial effect and is useful in the treatment of disorders or conditions/disorders that might be treated by DPP-IV inhibition.

Thus, the present invention relates to combinations, such as a combined preparation or pharmaceutical composition, respectively, comprising a DPP IV inhibitor or a pharmaceutically acceptable salt thereof, and i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or ii) omega-3 oils, or in any case a pharmaceutically acceptable salt thereof.

Preferably, the present invention relates to combinations, such as a combined preparation or pharmaceutical composition, respectively, comprising a DPP IV inhibitor or a pharmaceutically acceptable salt thereof, and i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or ii) omega-3 oils, or in any case a pharmaceutically acceptable salt thereof, and at least one additional pharmaceutically acceptable carrier.

Preferably the combination is a pharmaceutical composition or a combined pharmaceutical preparation.

In this pharmaceutical composition, the combination partners i.e. DPP IV inhibitor and , i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or ii) omega-3 oils, can be administered together, one after the other or separately in one combined unit dosage form or in two separate unit dosage forms. The unit dosage form may also be a fixed combination.

The term "at least one therapeutic agent" shall mean that in addition to the DPP IV inhibitor one or more, for example two, furthermore three, active ingredients as specified according to the present invention can be combined. Preferably one, two, three or four.

The term "DPP-IV" as used herein is intended to mean dipeptidyl peptidase IV, also known as CD26. DPP-IV, a serine protease belonging to the group of post-proline/alanine cleaving amino-dipeptidases, specifically removes the two N-terminal amino acids from proteins having proline or alanine in position 2. DPP-IV can be used in the control of glucose metabolism because its substrates include the insulinotropic hormones glucagon like peptide-1 (GLP-1) and gastric inhibitory peptide (GIP). GLP-1 and GIP are active only in their intact forms; removal of their two N-terminal amino acids inactivates them.

In vivo administration of synthetic inhibitors of DPP-IV prevents N- terminal degradation of GLP-1 and GIP, resulting in higher plasma concentrations of these hormones, increased insulin secretion and, therefore, improved glucose tolerance.

The term "DPP-IV inhibitor" is intended to indicate a molecule that exhibits inhibition of the enzymatic activity of DPP-IV and functionally related enzymes, such as from 1-100% or 20- 80% inhibition, and specially preserves the action of substrate molecules, including but not limited to GLP-1 , GIP, peptide histidine methionine, substance P, neuropeptide Y, and other molecules typically containing alanine or proline residues in the second amino terminal position. Treatment with DPP-IV inhibitors prolongs the duration of action of peptide substrates and increases levels of their intact, undegraded forms leading to a spectrum of biological activities relevant to the disclosed invention.

For that purpose, chemical compounds are tested for their ability to inhibit the enzyme activity of purified CD26/DPP-IV. Briefly, the activity of CD26/DPP-IV is measured in vitro by its ability to cleave the synthetic substrate Gly-Pro-p-nitroanilide (Gly-Pro-pNA). Cleavage of Gly-Pro-pNA by DPP-IV liberates the product p-nitroanilide (pNA), whose rate of appearance is directly proportional to the enzyme activity. Inhibition of the enzyme activity by specific enzyme inhibitors slows down the generation of pNA. Stronger interaction between an inhibitor and the enzyme results in a slower rate of generation of pNA. Thus, the degree of inhibition of the rate of accumulation of pNA is a direct measure of the strength of enzyme inhibition. The accumulation of pNA is measured spectrophotometrically. The inhibition constant, Ki, for each compound is determined by incubating fixed amounts of enzyme with several different concentrations of inhibitor and substrate.

In the present context "a DPP-IV inhibitor" is also intended to comprise active metabolites and prodrugs thereof, such as active metabolites and prodrugs of DPP-IV inhibitors. An active "metabolite" is an active derivative of a DPP-IV inhibitor produced when the DPP-IV inhibitor is metabolized. A "prodrug" is a compound that is either metabolized to a DPP-IV inhibitor or is metabolized to the same metabolite(s) as a DPP-IV inhibitor.

DPP-IV inhibitors are known in the art. For example, DPP-IV inhibitors are in each case generically and specifically disclosed e.g. in WO 98/19998.DE19616 486 A1 , WO 00/34241 , WO 95/15309, WO 01/72290, WO01/52825, WO 9310127, WO 9925719, WO 9938501 , WO 9946272, WO 9967278 and WO 9967279.

Preferred DPP-IV inhibitors are described in the following patent applications; WO 02053548 especially compounds 1001 to 1293 and examples 1 to 124, WO 02067918 especially compounds 1000 to 1278 and 2001 to 2159, WO 02066627 especially the described examples, WO 02/068420 especially all the compounds specifically listed in the examples I to LXIII and the described corresponding analogues, even preferred compounds are 2(28), 2(88), 2(119), 2(136) described in the table reporting IC50, WO 02083128 especially examples 1 to 13, US 2003096846 especially the specifically described compounds, WO 2004/037181 especially examples 1 to 33, WO 0168603 especially compounds of examples 1 to 109, EP1258480 especially compounds of examples 1 to 60, WO 0181337 especially examples 1 to 118, WO 02083109 especially examples 1A to 1 D, WO 030003250 especially compounds of examples 1 to 166, most preferably 1 to 8, WO 03035067 especially the compounds described in the examples, WO 03/035057 especially the compounds described in the examples, US2003216450 especially examples 1 to 450, WO 99/46272 especially compounds of claims 12, 14, 15 and 17, WO 0197808 especially compounds of claim 2, WO 03002553 especially compounds of examples 1 to 33, WO 01/34594 especially the compounds described in the examples 1 to 4, WO 02051836 especially examples 1 to 712, EP1245568 especially examples 1 to 7, EP1258476 especially examples 1 to 32, US 2003087950 especially the described examples, WO 02/076450 especially examples 1 to 128, WO 03000180 especially examples 1 to 162, WO 03000181 especially examples 1 to 66, WO 03004498 especially examples 1 to 33, WO 0302942 especially examples 1 to 68, US 6482844 especially the described examples, WO 0155105 especially the compounds listed in the examples 1 and 2, WO 0202560 especially examples 1 to 166, WO 03004496 especially examples 1 to 103, WO 03/024965 especially examples 1 to 54, WO 0303727 especially examples 1 to 209, WO 0368757 especially examples 1 to 88, WO 03074500 especially examples 1 to 72, examples 4.1 to 4.23, examples 5.1 to 5.10, examples 6.1 to 6.30, examples 7.1 to 7.23, examples 8.1 to 8.10, examples 9.1 to 9.30, WO 02038541 especially examples 1 to 53, WO 02062764 especially examples 1 to 293, preferably the compound of example 95 (2-{{3-(Aminomethyl)-4-butoxy-2-neopentyl-1-oxo-1 ,2 dihydro-6- isoquinolinyl}oxy}acetamide hydrochloride), WO 02308090 especially examples 1-1 to 1-109, examples 2-1 to 2-9, example 3, examples 4-1 to 4-19, examples 5-1 to 5-39, examples 6-1 to 6-4, examples 7-1 to 7-10, examples 8-1 to 8-8, examples 7-1 to 7-7 of page 90, examples 8-1 to 8-59 of pages 91 to 95, examples 9-1 to 9-33, examples 10-1 to 10-20, US 2003225102 especially compounds 1 to 115, compounds of examples 1 to 121 , preferably compounds a) to z), aa) to az), ba) to bz), ca) to cz) and da) to dk), WO 0214271 especially examples 1 to 320 and US 2003096857 and WO 2004/052850 especially the specifically described compounds such as examples 1 to 42 and compounds of claim 1, DE 102 56 264 A1 especially the described compounds such as examples 1 to 181 and the compounds of claim 5, WO 04/076433 especially the compounds specifically described, such as listed in table A, preferably the compounds listed in table B, preferably compounds I to XXXXVII, or compounds of claims 6 to 49, WO 04/071454 especially the specifically described compounds e.g. compounds 1 to 53 or compounds of tables Ia to If , or compounds of claims 2 to 55, WO 02/068420 especially the compounds specifically described, such as the compounds I to LXIII or Beispiele I and analogues 1 to 140 or Beispiele 2 and analogues 1 to 174 or Beispiele 3 and analogues 1 , or Beispiele 4 to 5, or Beispiele 6 and analogues 1 to 5, or Beispiele 7 and analogues 1-3, or Beispiele 8 and analogue 1 , or Beispiele 9, or Beispiele 10 and analogues 1 to 531 even preferred are compounds of claim 13, WO 03/000250 especially the compounds specifically described, such as the compounds 1 to 166, preferably compounds of examples 1 to 9, WO 03/024942 especially the compounds specifically described, such compounds 1 to 59, compounds of table 1 (1 to 68), compounds of claims 6, 7, 8, 9, WO 03024965024942 especially the compounds specifically described, such compounds 1 to 54, WO 03002593 especially the compounds specifically described, such compounds table 1 or of claims 2 to 15, WO 03037327 especially the compounds specifically described, such compounds of examples 1 to 209 WO 03/000250 especially the compounds specifically described, such as the compounds 1 to 166, preferably compounds of examples 1 to 9, WO 03/024942 especially the compounds specifically described, such compounds 1 to 59, compounds of table 1 (1 to 68), compounds of claims 6, 7, 8, 9, WO 03024965024942 especially the compounds specifically described, such compounds 1 to 54, Wo03002593 especially the compounds specifically described, such compounds table 1 or of claims 2 to 15, WO03037327 especially the compounds specifically described, such compounds of examples 1 to 209, WO0238541 , WO0230890.

WO 03/000250 especially the compounds specifically described, such as the compounds 1 to 166, preferably compounds of examples 1 to 9, WO 03/024942 especially the compounds specifically described, such compounds 1 to 59, compounds of table 1 (1 to 68), compounds of claims 6, 7, 8, 9, WO 03024965 especially the compounds specifically described, such compounds 1 to 54, WO 03002593 especially the compounds specifically described, such compounds table 1 or of claims 2 to 15, WO03037327 especially the compounds specifically described, such compounds of examples 1 to 209, WO0238541 especially the compounds specifically described, such compounds of examples 1 to 53, WO 03/002531 especially the compounds specifically described preferably the compounds listed on page 9 to 13, most preferably the compounds of examples 1 to 46 and even preferred compound of example 9, U.S. Patent No. 6,395,767 preferably compound of examples 1 to 109 most preferably compound of example 60,, U.S. application Serial No. 09/788,173 filed February 16, 2001 (attorney file LA50) especially the described examples, WO99/38501 especially the described examples, W099/46272 especially the described examples and DE19616 486 A1 _,

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especally val-pyr, val-thiazolidide, isoleucyl-thiazolidide, isoleucyl-pyrrolidide, and fumar salts of isoleucyl-thiazolidide and isoleucyl-pyrrolidide.

Further preferred DPP-IV inhibitors include the specific examples disclosed in United States Patent Numbers 6124305 and US 6107317, International Patent Applications, Publication Numbers WO 95153 09 and WO 9818763.

In each case in particular in the compound claims and the final products of the working examples, the subject matter of the final products, the pharmaceutical preparations and the claims are hereby incorporated into the present application by reference to these publications.

Published patent application WO 9819998 discloses N- (N'-substituted glycyl)-2-cyano pyrrolidines, in particular 1-[2-[5-Cyanopyridin-2-yl] amino]- ethylamino] acetyl-2-cyano- (S)- pyrrolidine (NVP-DPP728).

Published patent application WO 0034241 and published patent US 6110949 disclose N- substituted adamantyl-amino-acetyl-2-cyano pyrrolidines and W (substituted glycyl)-4-cyano pyrrolidines respectively. DPP-IV inhibitors of interest are specially those cited in claims 1 to 4. In particular these applications describe the compound 1-[[(3-Hydroxy-1-adamantyl) amino]acetyl]-2-cyano-(S)-pyrrolidine (also known as LAF237 or vildagliptin).

Published patent application WO 9515309 discloses amino acid 2- cyanopyrrolidine amides as inhibitors of DPP-IV Published patent application WO 9529691 discloses peptidyl derivates of diesters of alpha-aminoalkylphosphonic acids, particularly those with proline or related structures. DPP-IV inhibitors of interest are specially those cited in Table 1 to 8.

In WO 01/72290 DPP-IV inhibitors of interest are specially those cited in example 1 and claims 1 , 4, and 6.

WO 01/52825 specially discloses (S)-1 -{2-[5-cyanopyridin-2yl)amino]ethyl-aminoacetyl)-2- cyano- pyrrolidine or (S)-1 -[(3-hydroxy-1-adamantyl)amino]acetyl-2- cyano-pyrrolidine.

Published patent application WO 9310127 discloses proline boronic esters useful as DPP-IV inhibitors. DPP-IV inhibitors of interest are specially those cited in examples 1 to 19. Published patent application WO 9925719 discloses sulphostin, a DPP-IV inhibitor prepared by culturing a Streptomyces microorganism.

Published patent application WO 9938501 discloses N-substituted 4-8 membered heterocyclic rings. DPP-IV inhibitors of interest are specially those cited in claims 15 to 20. Published patent application WO 9946272 discloses phosphoric compounds as inhibitors of DPP-IV. DPP-IV inhibitors of interest are specially those cited in claims 1 to 23.

Published patent applications WO 9967278 and WO 9967279 disclose DPP-IV prodrugs and inhibitors of the form A-B-C where C is either a stable or unstable inhibitor of DPP-IV.

Other preferred DPP-IV inhibitors are the compounds of formula I, Il or III disclosed in the patent application WO 03/057200 on page 14 to 27. Most preferred DPP-IV inhibitors are the compounds specifically described on pages 28 and 29.

Any of the substances disclosed in the above mentioned patent documents, hereby included by reference, are considered potentially useful as DPP-IV inhibitors to be used in carrying out the present invention.

In a further preferred embodiment, the DPP-IV inhibitor is a N-peptidyl-O-aroyl hydroxylamine or a pharmaceutically acceptable salt thereof. Aroyl is, for example, naphthylcarbonyl; or benzoyl which is unsubstituted or mono- or disubstituted, for example, by lower alkoxy, lower alkyl, halogen or, preferably, nitro. The peptidyl moiety comprises preferably two α-amino acids, e.g. glycine, alanine, leucine, phenylalanine, lysine or proline, of which the one attached directly to the hydroxylamine nitrogen atom is preferably proline.

Preferably, the N-peptidyl-O-aroyl hydroxylamine is a compound of formula VII

wherein j is O, 1 or 2;

Rεi represents the side chain of a natural amino acid; and

Rε₂ represents lower alkoxy, lower alkyl, halogen or nitro; or a pharmaceutically acceptable salt thereof.

In a very preferred embodiment of the invention, the N-peptidyl-O-aroyl hydroxylamine is a compound of formula Vila

or a pharmaceutically acceptable salt thereof.

N-Peptidyl-O-aroyl hydroxylamines, e.g. of formula VII or Vila, and their preparation are described by H. U. Demuth et al. in J. Enzyme Inhibition 1988, Vol. 2, pages 129-142, especially on pages 130-132.

Preferred DPP-IV inhibitors are N-substituted adamantyl-amino- acetyl-2-cyano pyrrolidines, N (substituted glycyl)-4-cyano pyrrolidines, N- (N'-substituted glycyl)-2-cyanopyrrolidines, N- aminoacyl thiazolidines, N-aminoacyl pyrrolidines, L-allo-isoleucyl thiazolidine, L-threo- isoleucyl pyrrolidine, and L-allo-isoleucyl pyrrolidine, 1-[2-[(5-cyanopyridin-2-yl) amino] ethylamino] acetyl-2-cyano-(S)-pyrrolidine and pharmaceutical salts thereof.

Preferred DPP-IV inhibitors are those described by Mona Patel and col. (Expert Opinion Investig Drugs. 2003 Apr; 12(4):623-33) on the paragraph 5, especially P32/98, K-364, FE- 999011 , BDPX, NVP-DDP-728 and others, which publication is hereby incorporated by reference especially the described DPP-IV inhibitors.

Another preferred inhibitor is the compound BMS-477118 disclosed in WO 2001068603 or U.S. Patent No. 6,395,767 (compound of example 60) also known as is (1S,3S,5S)-2-[(2S)- 2-amino-2-(3-hydroxytricyclo[3.3.1.1³⁷]dec-1 -yl)-1 -oxoethyl]-2-azabicyclo[3.1.0]hexane-3- carbonitrile, benzoate (1 :1) as depicted in Formula M of the patent application WO 2004/052850 on page 2, and the corresponding free base, (IS,3S,5S)-2-[(2S)-2-amino-2- (3- hydroxy-tricyclo[3.3.1.1^3|7]dec-1 -yl)-1 -oxoethyl]-2-azabicyclo-[3.1.0]hexane-3-carbonitrile (M¹) and its monohydrate (M") as depicted in Formula M of the patent application WO 2004/052850 on page 3. The compound BMS-477118 is also known as saxagliptin.

Another preferred inhibitor is the compound GSK23A disclosed in WO 03/002531 (example 9) also known as (2S.4S)- 1- ((2R)-2-Amino-3-[(4-methoxybenzyl)sulfonyl]-3- methylbutanoyl)-4-fluoropyrrolidine-2-carbonitrile hydrochloride.

FE-999011 is described in the patent application WO 95/15309 page 14, as compound No. 18. P32/98 or P3298 (CAS number: 251572-86-8) also known as 3-[(2S,3S)-2-amino-3-methyl- 1-oxopentyl]thiazolidine can be used as 3-[(2S,3S)-2-amino-3-methyl-1- oxopentyl]thiazolidine and (2E)-2-butenedioate (2:1) mixture such as shown below

and is described in WO 99/61431 and also in Diabetes 1998, 47, 1253-1258, in the name of Probiodrug, as well as the compound P93/01 described by the same company.

Other very preferred DPP-IV inhibitors of the invention are described in the International patent application WO 02/076450 (especially the examples 1 to 128) and by Wallace T. Ashton (Bioorganic & Medicinal Chemistry Letters 14 (2004) 859-863 ) especially the compound 1 and the compounds listed in the tables 1 and 2. The preferred compound is the compound 21 e (table 1) of formula :

Other preferred DPP-IV inhibitors are described in the patent applications WO 2004/037169 especially those described in the examples 1 to 48 and WO 02/062764 especially the described examples 1 to 293, even preferred are the compounds 3-(aminomethyl)-2- isobuthyl-1-oxo-4-phenyl-1 ,2-dihydro-6-isoquinolinecarboxamide and 2-{[3-(aminomethyl)-2- isobuthyl-4-phenyl-1-oxo-1 ,2-dihydro-6-isoquinolyl]oxy}acetamide described on page 7 and also in the patent application WO2004/024184 especially in the reference examples 1 to 4. Other preferred DPP-IV inhibitors are described in the patent application WO 03/004498 especially examples 1 to 33 and most preferably the compound of the formula

MK-0431 described by the example 7 and also known as MK-0431 or Sitagliptin.

Preferred DPP-IV inhibitors are also described in the patent application WO 2004/037181 especially examples 1 to 33 and most preferably the compounds described in the claims 3 to 5.

Other DPP-IV inhibitors are (2S)-1- { (2-(5-Methyl-2-phenyl-oxazol-4-yl)-ethylamino)-acetyl}- pyrrolidine-2-carbonitrile, or (2S)-1-{ (1,1-Dimethyl-3-(4-pyridin-3-yl-imidazol-1-yl)- propylamino)-acetyl}-pyrrolidine-2 -carbonitrile, and pharmaceutically acceptable salts thereof. (2S)-1- { (2-(5-Methyl-2-phenyl-oxazol-4-yl)-ethylamino)-acetyl}-pyrrolidine-2- carbonitrile is preferably used in form of the mesylate salt. These compounds and methods for their preparation have been disclosed and described in WO 03/037327.

Other DPP-IV inhibitors are (S)-1-( (2S.3S, 11 bS)-2-Amino-9, 10-dimethoxy-1 , 3,4,6,7, 11b- hexahydro-2H-pyrido (2,1 -a) isoquinolin-3-yl)-4-fluoromethyl-pyrrolidin-2-one, or (S₁S₁S₁S)-I- (2-Amino-9, 10-dimethoxy-1 ,3,4,6,7,1 11 b-hexahydro-2H-pyrido (2,1 -a) isoquinolin-3-yl)-4- methyl-pyrrolidin-2-one, and pharmaceutically acceptable salts thereof. (S)-1-( (2S.3S, 11bS)-2-Amino-9, 10-dimethoxy-1 , 3,4,6,7, 11b-hexahydro-2H-pyrido (2,1 -a) isoquinolin-3-yl)- 4-fluoromethyl-pyrrolidin-2-one and pharmaceutically acceptable salts thereof is preferred.

The compounds of formula (II) and methods for their preparation have been described in WO 2005/000848.

Preferred DPP-IV inhibitors are N-substituted adamantyl-amino- acetyl-2-cyano pyrrolidines, N (substituted glycyl)-4-cyano pyrrolidines, N- (N'-substituted glycyl)-2-cyanopyrrolidines, N- aminoacyl thiazolidines, N-aminoacyl pyrrolidines, L-allo-isoleucyl thiazolidine, L-threo- isoleucyl pyrrolidine, and L-allo-isoleucyl pyrrolidine, 1-[2-[(5-cyanopyridin-2-yl) amino] ethylamino] acetyl-2-cyano- (S)-pyrrolidine , MK-431 and pharmaceutical salts thereof.

Preferred DPP-IV inhibitors are selected from [S]-1-[2-(5-cyano-2- pyridinylamino)ethylamino]acetyl-2-pyrolidine carbonitrile monohydrochloride, (S)-1-[(3- hydroxy-1-adamantyl)amino]acetyl-2-cyano-pyrrolidine and L-threo-isoleucyl thiazolidine (compound code according to Probiodrug: P32/98 as described above), MK-0431 , 3- (aminomethyl)-2-isobuthyl-1-oxo-4-phenyl-1 ,2-dihydro-6-isoquinolinecarboxamide and 2-{[3- (aminomethyl)-2-isobuthyl-4-phenyl-1 -oxo-1 ,2-dihydro-6-isoquinolyl]oxy}acetamide and optionally pharmaceutical salts thereof.

Especially preferred are 1-{2-[(5-cyanopyridin-2-yl) amino] ethylamino} acetyl-2 (S)- cyano- pyrrolidine dihydrochloride (DPP728) (also named [S]-1-[2-(5-cyano-2- pyridinylamino)ethylamino]acetyl-2-pyrolidine carbonitrile monohydrochloride), of formula

especially the dihydrochloride and monohydrochloride thereof, and 1-[(3-hydroxy-1-adamantyl) amino] acetyl-2-cyano-, (S) (also named (S)-1-[(3-hydroxy-

1-adamantyl)amino]acetyl-2-cyano-pyrrolidine, LAF237 or vildagliptin) of formula

and L-threo-isoleucyl thiazolidine (compound code according to Probiodrug: P32/98 as described above), MK-0431 , (2S)-1- { (2-(5-Methyl-2-phenyl-oxazol-4-yl)-ethylamino)-acetyl}- pyrrolidine-2-carbonitrile, or (2S)-1-{ (1 ,1-Dimethyl-3-(4-pyridin-3-yl-imidazol-1-yl)- propylamino)-acetyl}-pyrrolidine-2 -carbonitrile, (S)-1-( (2S.3S, 11 bS)-2-Amino-9, 10- dimethoxy-1 ,3,4,6,7,11 b-hexahydro-2H-pyrido (2,1 -a) isoquinolin-3-yl)-4-fluoromethyl- pyrrolidin-2-one, or (S,S,S,S)-1-(2-Amino-9, 10-dimethoxy-1 , 3,4,6,7,1 11 b-hexahydro-2H- pyrido (2,1 -a) isoquinolin-3-yl)-4-methyl-pyrrolidin-2-one, GSK23A, saxagliptin, 3- (aminomethyl)-2-isobuthyl-1-oxo-4-phenyl-1 ,2-dihydro-6-isoquinolinecarboxamide and 2-{[3- (aminomethyl)-2-isobuthyl-4-phenyl-1 -oxo-1 ,2-dihydro-6-isoquinolyl]oxy}acetamide and optionally pharmaceutical salts thereof.

DPP728 and vildagliptin are specifically disclosed in Example 3 of WO 98/19998 and Example 1 of WO 00/34241 , respectively. The DPP-IV inhibitor P32/98 (see above) is specifically described in Diabetes 1998, 47, 1253-1258. DPP728 and LAF237 can be formulated as described on page 20 of WO 98/19998 or in WO 00/34241 or in the International Patent Application No. EP2005/000400 (application number).

Especially preferred are orally active DPP-IV inhibitors.

Any of the substances disclosed in the above mentioned patent documents or scientific publications, hereby included by reference, are considered potentially useful as DPP-IV inhibitors to be used in carrying out the present invention.

Omega-3 oils, omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides are well known in the art and are properly described in e.g. the patent application WO 2006/017698 which is incorporated into the present application by reference to this application.

"Fatty acids" are an important component of nutrition. Fatty acids (also described as "free acids" or "free fatty acids") are carboxylic acids and are classified based on the length and saturation characteristics of the carbon chain. Short chain fatty acids have 2 to about 5 carbons and are typically saturated. Medium chain fatty acids; have from about 6 to about 14 carbons and are also typically saturated. Long chain fatty acids have from about 15 to 24 or more carbons and may also be saturated or I unsaturated. In longer fatty acids there may be one or more points of unsaturation, giving rise to the terms "monounsaturated" and "polyunsaturated", respectively. Long chain polyunsaturated fatty acids (LCPs or LC-PUFAs) having 20 or more carbons are used in the instant invention.

"Long chain" mono-, di-, ki-glycerides, esters, fatty acids, etc. are deemed as having about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or more carbons and may] also be saturated or unsaturated. "Medium chain" mono-, di-, tri-glycerides, esters, fatty acids, etc. are deemed as having about 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbons and may also be saturated or unsaturated. "Short chain" mono-, di-, ki-glycerides, esters, ] fatty acids, etc. are defined as having about 2, 3, 4, or 5, carbons and may also be saturated or unsaturated.

"Mono-diglyceride" and "mono-diglycerides" refer to a mixture or [ mixtures comprising both monoglycerides and diglycerides. A non-limiting example of a mono-diglyceride is Capmul MOM, which comprises a mixture of caprylic and capric fatty acids in the form of monoglycerides and diglycerides. Certain mixtures of monoglycerides and diglycerides may be specifically stated as mono-diglycerides according to the present invention. Mono- diglycerides can comprise other species such as, for example, triglycerides and glycerol.

LC-PUFAs are categorized according to the number and position of double bonds in the fatty acids according to an accepted nomenclature that is well known to those of ordinary skill in the art. There are two series or families of LC PUFAs, depending on the position of the double bond closest to the methyl end of the fatty acid: the n-3 series contains a double bond at the third carbon, while the n-6 series has no double bond until the sixth carbon. Thus, arachidonic acid (AA or ARA) has a chain length of 20 carbons and 4 double bonds beginning at the sixth carbon. As a result, it is referred to as "20:4 n-6". Similarly, docosahexaenoic acid (DHA) has a chain length of 22 carbons with 6 double bonds beginning with the third carbon from the methyl end and is thus designated "22:6 n-3". Another important LC-PUFA is eicosapentaenoic acid (EPA) which is designated (20:5 n-3). The terms "n-3" and "omega-3" are used interchangeably.

The biosynthetic pathways for AA (n-6 series) and DHA (n-3 series) from their respective (A 18 precursors are distinct, but share elongation and desaturation steps and are well understood. Thus, other important LCPs are the C18 fatty acids that are precursors in these biosynthetic pathways, for example, linoleic (18:2 n-6) and I gamma-linolenic (18:3 n-6) acids in the n-6 pathway, and alpha-linolenic (18:3 n-3) and stearidonic (18:4 n-3) in the n-3 pathway.

Fatty acids are often found in nature as acyl radicals esterifed to alcohols. A glyceride is such an ester of one or more fatty acids with glycerol (1 ,2,3 propanetriol). If only one position of the glycerol backbone molecule is esterifed with a fatty acid, a "monoglyceride" is produced; if two positions are esterified, a "diglyceride" is produced; and if all three positions of the glycerol are esterified with fatty acid a "triglyceride" or "triacylglycerol" is produced. A glyceride is called "simple" if all esterifed positions contain the same fatty acid; or "mixed" if different] fatty acids are involved. A phospholipid is a special type of diglyceride, wherein the third position on the glycerol backbone is bonded to a nitrogen containing compound such as choline, serine, ethanolamine, inositol, etc., via a phosphate ester. Triglycerides [ and phospholipids are often classifed as long chain (from about 15 to 24 or more carbons) or medium chain (from about 6 to about 14 carbon), according to the fatty acids attached thereto.

Typically commercially available monoglycerides contain varying amounts of di- and triglycerides in addition to their monoglyceride content. For example, a monoglyceride (e.g., Akoline, by Karlshamns AB, Sweden) can comprise about 50-65 % monoglyceride, 25-35 % diglyceride, and up to 5 % triglycerides.

The "essential fatty acids" (EFAs) are of two types, the n-3 (or omega-3) series derived from alpha-linolenic acid and the n-6 (or omega-6) series derived from linoleic acid.

An "omega-3 fatty acid" is a n-3 polyunsaturated long-chain fatty acids (n-3 PUFA) and is defined to include any carboxylic acid having at least 15 carbon atoms and having at least 3 non-conjugated cis-unsaturated bonds, the distal one of which from the methyl end of the fatty acid chain being located between the third and fourth carbon atoms. The omega-3 fatty acids therefore include Cry -C24 alkanoic acids comprising 5-7 double bonds, wherein the last double bond is located between the third and fourth carbon atom from the methyl end of the fatty acid chain.

Examples of omega-3 fatty acids include stearidonic acid (ST?A, C18:4), eicosatetraenoic acid (ETA, C20:4), eicosapentaenoic acid (EPA, C20:5), ; docosapentaenoic acid (DPA, C22:5), and docosahexaenoic acid (DIVA, C22:6). For the purpose ofthe invention, alpha- linolenic acid (ALA, C18:3) is considered an I omega-3 fatty acid. Terms such as "EPA" and "DIVA" denote species of omega-3 oil and do not describe whether such oils exist as, for example, triglycerides, diglycerides, monoglycerides, free acids, esters, or salts.

Omega-3 fatty acids include synthetic or naturally occurring omega-3 fatty acids, such as those found in fish oil, e.g., marine mammal (e.g., seal) fat, cod liver oil, walnuts and walnut oil, wheat germ oil, rapeseed oil, soybean lecithin, ] soybeans, tofu, common beans, butternuts, seaweed and flax seed oil. An omega-3 fatty acid may also be derived from genetically engineered sources such as transgenic plants.

See, e.g., Frasier, et al., Nat Biotechnol. 2004 May 16. ] An "omega-3 oil" or "omega-3" is any oil comprising a source of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters, or omega-3 mono-, di-, or triglycerides, such as fish oil, e.g., marine mammal (e.g., seal) fat, cod liver oil, walnuts and walnut oil, wheat germ oil, rapeseed oil, soybean lecithin derived oils, soybean derived oils, tofu derived oils, common bean derived oils, butternut derived oils, seaweed derived oils, flax-borage oil, and flax seed oil. The Epax (Pronova Biocare AS) brand of omega-3 oils are preferred. Other omega-3 oils which can be used in making pharmaceutical compositions of the invention include, but are not limited to, the omega-3 oil marketed under the tradename Omegabrite (Omega Natural Science) and Epanova_ (Tillotts Pharma AG). Certain mixtures of esters, fatty acids, and/or mono- di- triglycerides may be specifically stated as oils according to the present invention. For example, a mixture consisting of omega-3 esters and fatty acids may be considered an omega-3 oil according to the present invention. In addition, one or more components may be specifically excluded from an omega-3 oil according to the present invention. For example, an omega-3 oil may specifically exclude esters, fatty acids, and/or mono- di- triglycerides according to the present invention. As such, a composition consisting of omega- 3 esters, for example, is an omega-3 oil according to the present invention.

An "omega-3 alkyl ester" may be formed by transesterifcation of an omega-3 oil and an alcohol (preferably methanol or ethanol) and either an acid or reducing agent. Because formation of lower alkyl esters is generally preferred, the alcohol preferably is a lower alkyl alcohol containing from I to 6 carbon atoms. More preferably, the alcohol is methanol (which reacts with glycerides to form methyl esters of the fatty acid residues) or ethanol (which reacts with glycerides to form ethyl esters of the fatty acid residues). Most preferably, the alcohol is ethanol. Omega-3 alkyl ester which can be used in making pharmaceutical compositions/combinations of the invention include, but are not limited to, the omega-3 acid ethyl esters marketed under the trade name OMACOR ® by Norsk Hydro.

The term "E463808" is used to described an omega-3 oil which has a composition comprising 46 % EPA, 38 % DHA, and 8 % other omega-3 oils (mass percent) where the EPA, DHA, and other omega-3 oils are ethyl esters.

The term "E681010"is used to describe an omega-3 oil which has a composition comprising 67.8 percent EPA (mg/g), 9.9 percent DHA (mg/g), and about 9.6 percent other omega-3 oils (mg/g), where the EPA, DHA, and other omega-3 oils are ethyl esters. Omega-3 alkyl esters include the ethyl esters of EPA and DHA. The E463808, OMEGA- 3/90 (K D Pharma), and lncromega (CrodalBioriginal) omega-3 ethyl esters are several exemplary omega-3 alkyl esters.

"Alkyl" means a straight chain or branched, saturated or unsaturated alkyl, cyclic or non- cyclic hydrocarbon having from 1 to 10 carbon atoms.

Representative saturated straight chain alkyls include methyl, ethyl, n- propyl, n-butyl, n- pentyl, n-hexyl, and the like; while saturated branched alkyls include isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, and the like. Unsaturated alkyls contain at least one double or triple bond between adjacent carbon atoms (also referred to as an "alkenyl" or "alkynyl", respectively). Representative straight chain and branched] alkenyls include ethylenyl, propylenyl, 1-butenyl, 2-butenyl, isobutylenyl, 1-pentenyl, 2- pentenyl, 3-methyl-1-butenyl, 2- methyl-2-butenyl, 2,3-dimethyl-2-butenyl, and the like; while representative straight chain and branched alkynyls include acetylenyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2- pentynyl, 3-methyl- 1 butynyl, and the like. Representative saturated cyclic alkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like; while unsaturated cyclic alkyls include cyclopentenyl and cyclohexenyl, and the like. Cycloalkyls are also referred to herein as "carbocyclic" rings systems, and include bi- and tri-cyclic ring systems having from 8 to 14 carbon atoms such as a cycloalkyl (such as cyclopentane or cyclohexane) fused to one or more aromatic (such as phenyl) or non-aromatic (such as cyclohexane) carbocyclic rings. "Alkenyl" can be used in the context of omega-3 formulations to refer to unsaturation.

"omega-3 esters" may be formed by transesterification of an omega-3 oil and an alcohol and either an acid or reducing agent.

Acid-catalyzed transesterification may be carried out, for example, by incubating a triglyceride at from about 0^cC to about ISO⁰C in a mixture containing the alcohol and an acid (e.g., HCI), preferably under a non- oxidizing atmosphere and in the absence of water. In one embodiment, the triglyceride/acid/alcohol mixture is refluxed for at least about 2 hours. In another embodiment, the triglyceride/acid/alcohol mixture is maintained at from about 0⁰C to about 50⁰C overnight. Methanol may be used to: form methyl esters, and ethanol may be used to form ethyl esters. Because acid catalyzed transesterification is typically reversible, the alcohol preferably is present in a large excess so that the reaction proceeds essentially to completion. Preferably, the triglyceride concentration in the alcohol/acid mixture is from about 0.1 to about 15% by weight, and most preferably about 3% by weight. If the acid is HCI, the concentration of HCI in the alcohol/HCI mixture preferably is from about 4 to about 15% by weight, and most preferably about 10% by weight. Such a mixture may be prepared by various methods known in the art, such as bubbling dry gaseous hydrogen chloride into dry ethanol, or adding 1 mL of acetylchloride to each 10 mL of alcohol (to form approximately 10% by weight HCI in alcohol).

Although HCI is most preferred, other acids may alternatively be used.

One such acid is sulfuric acid, which typically is used at a concentration of from about 0.5 to about 5% by weight in the alcohol. It should be noted, however, that because sulfuric acid is a strong oxidizing agent, it preferably is not used with long reflux times (i.e., greater than about 6 hours), at high concentrations (i.e., greater than about 5% by weight), or at high temperatures (i.e., greater than 150° C). Another example of a suitable acid is boron trifluoride, which preferably is used at a concentration of from about 1 to about 20% by weight in the alcohol. Boron trifluoride, however, is less preferred than HCI because boron trifluoride has a greater tendency to produce undesirable byproducts.

In hale-catalyzed transesterification, the omega-3 oil is transesterified by an alcohol in the presence of a basic catalyst. In this instance, the base may be, for example, sodium methoxide, potassium methoxide, elemental sodium, sodium hydroxide, or potassium hydroxide. Preferably, the volumetric ratio of omega-3 oil to the base/alcohol mixture is at least about 1 :1 , and most preferably about 1 :2. The concentration of the base in the alcohol preferably is from about 0.1 to about 2 M. The base-catalyzed transesterifcation reaction can be conducted at room temperature (i.e., at a temperature of from about 20° to about 25⁰C) for from about 6 to about 20 hours.

Alternatively, the base-catalyzed transesterification reaction is conducted at a temperature greater than room temperature.

The glyceride/alcohol/catalyst solution preferably is heated to a temperature of at least about 40⁰C, more preferably from about 70 to about 150⁰C, and most preferably at about 100⁰C. The solution can be heated using a reflux condenser so that the reaction mixture may be heated to temperatures above the boiling point of one or more components in the mixture without losing the components into the vapor phase (i.e., when the components vaporize, they rise into the reflux condenser which has a cooler temperature, thereby causing the vapor to condense into a liquid and flow back into the liquid mixture). During the transesterification reaction, the reacting mixture is preferably placed under a non- oxidizing atmosphere, such as an atmosphere consisting essentially of a noble gas, N₂, or a combination thereof. Use of such an atmosphere is particularly preferred if the transesterification reaction is conducted over a period of time exceeding about 10 minutes. An oil- soluble antioxidant (e.g., ascorbyl palmitate or propyl gallate) may also be added to the reacting mixture to prevent auto-oxidation, and is particularly preferred where a non- oxidizing atmosphere is not used.

Omega-3 alkyl esters include the ethyl esters of EPA and DEJA. The 6463808, OMEGA- 3/90 (K D Pharma), and lncromega (CrodalBioriginal) omega-3 ethyl esters are several exemplary omega-3 alkyl esters.

Pharmaceutical compositions and medicaments may be described as mixtures of two or more components "by volume," which is herein defined as the volume due to one component divided by the volume of all components of the composition. This ratio may be converted to or reported as a percentage of the total composition volume. Such a quantity may also be indicated by "v/v" or "percent v/v." Similarly, the phrases "by weight" and "by mass" describe the weight or mass due to one component divided by the weight or mass of all components of the composition. This ratio may be converted to or reported as a percentage of the total composition weight or mass. Such a quantity may also be indicated by "w/w", "mass percent," or percent w/w."

The terms "pharmaceutical composition" and "formulation" are used! interchangeably throughout the specification and claims.

Oil purity is also an important aspect of the present invention. Oil purity is defined as a percentage (e.g., by volume or by weight) of one component with respect to the entire oil composition. Several examples of oil components include, but are not limited to, monoglycerides, diglycerides, triglycerides, free acids, esters, and derivatives, precursors, and salts thereof. For example, an ester oil with a purity of 95 percent by weight comprises at least 95 percent esters. The remaining percentage may comprise free acids, mono- di- and/or triglycerides, or other components. As another example, an omega-3 ester oil with a purity of 90 percent by weight comprises at least percent omega-3 esters and the remaining percentage can comprise any one or more; of other oil components. A mixture of species of one component (e.g., C₈ and Ci₀: esters) need not be discerned in the determination of purity. However, a distinction of specific species within a component (e.g., C₈ and C₁₀ esters) can also be included in specific embodiments of the present invention.

According to the present invention, omega-3 oils with a purity greater than about 85 percent, 90 percent, 91 percent, 92 percent, 93 percent, 94 percent, 95 percent, 96 percent, 97 percent, 98 percent, 99 percent or more are preferred. Omega-3 oils with a high purity of omega-3 esters are preferred. According to the present invention, omega-3 oils with a high purity comprise greater than about 85 percent, 90 percent, 91 percent, 92 percent, 93 percent, 94 percent, 95 percent, 96 percent, 97 percent, 98 percent, 99 percent or more of one component by weight or by volume.

Preferred omega-3 esters include, but are not limited to, EPA and DHA. More preferred omega-3 esters include omega-3 ethyl esters.

Oil composition is another important aspect of the present invention.

Oil composition can be described as both the species and the components of an oil.

Species include specific omega-3 oils such as, but not limited to, EPA, DHA, linoleic acid, linolenic acid, etc. Components include, but are not limited to, monoglycerides, diglycerides, triglycerides, free acids, esters, and derivatives, precursors, and salts thereof. For example, E463808 comprises about 46 % EPA and about 38 % DHA (mass percent) as ethyl esters. The remaining portion consists essentially of omega-3 oils other than EPA and DHA and other non-omega-3 oils. Other commercially available omega-3 oils contain higher or lower levels of total EPA and DHA as components such as monoglycerides, diglycerides, triglycerides, esters, free acids, etc. or mixtures thereof. Omega-3 oils with a composition comprising a mass percent of EPA and DHA equal to or greater than about 55 percent are preferred. Omega-3 oils; with a composition comprising a mass percent of EPA and DHA equal to or greater than about 75 percent are more preferred. Omega-3 oils with a composition comprising a mass percent of EPA and DHA equal to or greater than about 80 percent are most preferred.

Mixtures of omega-3 alkyl esters with other forms of omega-3 oil (e.g., fatty acids, triglycerides) are included, according to the present invention. Oils containing highly pure or pure alkyl esters are included in the present invention.

In another embodiment, the purity of omega-3 esters or omega-3 alkyl esters is at least about 50 percent by weight, at least about 60 percent by weight, at least about 70 percent by weight, at least about 75 percent by weight, at least about 80 percent by weight, or at least about 85 percent by weight. In another embodiment, the purity of omega-3 esters or omega- 3 alkyl esters is about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99 percent or more by weight. In another embodiment, the purity of omega-3 esters or omega-3 alkyl esters is between about 25 and about 100 percent by weight, between about 40 and about 100 percent by weight, between about and about 100 percent by weight, between about 60 and about 100 percent by weight, between about 70 and about 100 percent by weight, between about 75 and about 100 percent by weight, between about 75 and about 95 percent by weight, between about 75 and about 90 percent by weight, or between about 80 and about 85 percent by weight. In another embodiment, the purity of omega-3 esters or omega- 3 alkyl esters is about 100 percent by weight, about 99 percent by weight, about 96 percent by weight, about 92 percent by weight, about 90 percent by weight, about 85 percent by weight, about 80 percent by weight, about 75 percent by weight, about 70 percent by weight, about 65 percent by weight, about 60 percent by weight, about 55 percent by weight, or about 50 percent by weight.

In another embodiment, the oil composition comprising EPA and DHA is at least about 50 percent by weight, at least about 60 percent by weight, at least about 70 percent by weight, at least about 75 percent by weight, at least about 80 percent by weight, or at least about 84 percent by weight. In another embodiment, the oil composition comprising EPA and DHA is about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95 percent by weight. In another embodiment, the oil composition comprising EPA and DHA is between about 25 and about 95 percent by weight, between about 40 and about 95 percent by weight, between about 50 and about 95 percent by weight, between about 60 and about 95 percent by weight, between about 70; and about 95 percent by weight, between about 75 and about 95 percent by weight, : between about 75 and about 90 percent by weight, between about 75 and about 8S percent by weight, or between about 80 and about 85 percent by weight. In another embodiment, the oil composition comprising EPA and DHA is about 99 percent by weight, about 96 percent by weight, about 92 percent by weight, about 90 percent by weight, about 84 percent by weight, about 80 percent by weight, about 75 percent by weight, about 70 percent by weights about 65 percent by weight, about 60 percent by weight, about 55 percent by weight, or about 50 percent by weight.

In another embodiment, the omega-3 ester or omega-3 alkyl ester has about a 23:19 ratio of EPA:DHA, about a 75:11 ratio of EPA:DHA, about a 95:1 ratio of EPA:DHA, about a 9:2 ratio of EPA: DHA, about a 10:1 ratio of EPA: DHA, about a 5:1 ratio of EPA: DHA, about a 3:1 ratio of EPA:DHA, about a 2:1 ratio of EPA:DHA, about a 1 :1 ratio of EPA:DHA, about a 1 :2 ratio of EPA:DHA, about a 1 :3 ratio of EPA:DHA, or about a 1 :5 ratio of EPA:DHA. In another embodiment, the omega-3 ester or omega-3 alkyl ester has about a 95:1 ratio of EPA: DHA, about a 75:1 ratio of EPA: DHA, about a 50:1 ratio of EPA: DHA, about a 25:1 ratio of EPA: DHA, about a 20:1 ratio of EPA: DHA, about a 15:1 ratio of EPA: DHA, about a 10:1 ratio of EPA:DHA, about a 7.5:1 ratio of EPA:DHA, about a 5:1 ratio of EPA:DHA, about a 14:1 ratio of EPA:DHA, about a 3:1 ratio of EPA:DHA, about a 2:1 ratio of EPA:DHA, about a 1.5: 1 ratio of EPA:DHA, about a 1 : 1 ratio of EPA:DHA, about a 1 : 1.5 ratio of EPA:DHA, about a 1 :2 ratio of EPA:DHA, about a 1 :3 ratio of EPA:DHA, or about a I 1 :5 ratio of EPA:DHA. In another embodiment, the omega-3 ester or omega-3 alkyl ester has from about a 95:1 ratio to about a 1 :5 ratio of EPA:DHA, from about a 50:1 ratio to about a 1 :1 ratio of EPA:DHA, from about a 25:1 ratio to about a 1 :1 ratio of EPA: DHA, from about a 10: 1 ratio to about a 1 : 1 ratio of EPA:DHA, from about a 5:1 ratio to about a 1 :1 ratio of EPA:DHA, from about a 3:1 ratio to about a 1 :1 ratio of EPA:DHA, from about a 2:1 ratio to about a 1 :1 ratio of EPA:DHA, or from about a i 1.5:1 ratio to about a 1:1 ratio of EPA:DHA. In another embodiment, the omega-3 ester or omega-3 alkyl ester has at least about a 1 :5 ratio of EPA:DHA, at least about a 1 : 1 ratio of EPA:DHA, at least about a 1.5: 1 ratio of EPA:DHA, at least about a 2: 1 ratio of EPA: DHA, at least about a 3:1 ratio of EPA: DHA, at least about a 5:1 ratio of EPA: DHA, or at least about a 10:1 ratio of EPA: DHA.

Example of preferred specific ratio, composition, or purity of omega-3 oil is for example an omega-3 oil comprising 90 percent (w/w) omega-3 ethyl esters with 46 percent EPA and 38 percent DHA (e.g., OMACOR).

Other preferred commercially available omega-3 oils are for example those available from Croda International t (England) and Pronova Biocare (Norway).

The dosage of omega-3 oil administered will also be generally dependent upon the health of the subject being treated, the extent of treatment desired, the nature and kind of concurrent therapy, if any, and the frequency of treatment and nature of the effect desired. A therapeutically acceptable daily dosage of omega-3 oil has been recommended or considered via several national and international groups including, but not limited to, the American Heart Association (AMA) and the International Society for the Study of Fatty Acids and lipids (ISSFAL).

Table 1 includes daily dosage amounts of omega-3 as considered/recommended via several organizations.

In general, the dosage of the omega-3 oils is generally in the range of from about 0.001 to about 100 mg/kg body weight of the subject per day, preferably from about 0.1 to about 50 mg/kg body weight of the subject per day, administered as a single or divided dose. However, some variability in the general dosage range may also be required depending upon the age, weight, and species of the patient, the intended route of administration, and the progress and degree of severity of the disease or condition being treated.

The dosage of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, administered will also be generally dependent upon the health of the subject being treated, the extent of treatment desired, the nature and kind of concurrent therapy, if any, and the frequency of treatment and nature of the effect desired.

Daily dosages of omega-3 oil required in practicing the method of the present invention will vary depending upon, for example the mode of administration and the severity of the condition to be treated. An indicated daily dose is in the range of from about 100 to about 5000 mg, e.g. from 100 to 4000 mg or e.g. from 200 to 4000 mg or e.g. from 500 to 4000 mg , or e.g. between 500 to 2000 mg of active agent for oral use, conveniently administered once or in divided dosages.

Preferred omega-3 oils are selected from Epax® , Omegabrite®, Epanova®, E463808, E681010, Omacor®, OMEGA- 3/90, Incromega, Epadel®, Seacor®, Esapent®, Eskim®, Liposol® Medtech Pharma Sverige AB.

Epadel: A preferred EPA formulation CINN: lcosapent) used in the present invention is Epadel (Mochida; Tokyo, Japan), an ethical drug containing highly purified (>98%) fish- derived ethyl EPA, which is currently indicated in Japan for the treatment of arteriosclerosis and hyperlipidemia. Preferably the daily dosage is between 500 and 4000 mg of EPA preferably e.g. 1800 mg/day.

Eskimo-3® brand Fish Oil provides omega-3 fatty acids including 645-830 mg EPA (eicosapentaenoic acid) and 380-540 mg DHA (docosahexaenoic acid) 4.6 g - Serving Size: 1 teaspoon (5 mL) Amount/Serving

Eskimo Kids 105 ml ® : Calories 40 , Calories from Fat 40 , Total Fat 5 g , Saturated Fat 0.5 g , Vitamin D (as Cholecalciferol) 100 IU , Omega-3 Fatty Acids 800 mg , EPA (eicosapentaenoic acid) 270 mg , DHA (docosahexaenoic acid) 180 mg , Canola Oil Containing: (Oleic Acid, ϋnoleic Acid, Alpha-Linolenic Acid) 2g - Serving Size: 1 Teaspoon (5 mL) Amount/Serving.

Seacor^®, Esapent^®, Eskim^® are n-3 PUFA and administered preferably at a dose of 500 to 3000 mg a day preferably e.g. 1 g once a day. Eskim® contains omega-3 triglycerides (DHA, EPA) and is marketed by Sigma Tau S.p.A in Italy.

Epax® (Pronova Biocare AS): Epax is a brand of different Omega-3 oils. E.g. Epax Omega-3 Joint Formula contains is in the form of a easy-to-swallow softgels containing 1000 mg of Omega 3 fish oil, which contains around 375 mg of EPA and around 67 mg of DHA. Other preferred Epax products are Epax 6000 TG, Epax 6000 EE, Epax 5500 TG, Epax 5500 EE.

Daily dosages of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, required in practicing the method of the present invention will vary depending upon, for example the mode of administration and the severity of the condition to be treated. An indicated daily dose is in the range of from about 100 to about 5000 mg, e.g. from 100 to 4000 mg or e.g. from 200 to 4000 mg or 500 to 4000 mg or e.g. 500 to 2000 mg of active agent for oral use, conveniently administered once or in divided dosages.

Any of the above described combination which is intended for a daily administration.

The corresponding active ingredients or a pharmaceutically acceptable salt thereof may also be used in form of a solvate, such as a hydrate or including other solvents, used for crystallization.

The compounds to be combined can be present as pharmaceutically acceptable salts. If these compounds have, for example, at least one basic center, they can form acid addition salts. Corresponding acid addition salts can also be formed having, if desired, an additionally present basic center. The compounds having an acid group (for example COOH) can also form salts with bases.

All of these marketed products may be utilized in as such for combination therapy according to the present invention.

The structure of the active agents identified by generic or trade names may be taken from the actual edition of the standard compendium "The Merck Index" or from databases, e.g. Patents International (e.g. IMS World Publications). The corresponding content thereof is hereby incorporated by reference. Any person skilled in the art is fully enabled to identify the active agents and, based on these references, likewise enabled to manufacture and test the pharmaceutical indications and properties in standard test models, both in vitro and in vivo.

All the more surprising is the experimental finding that the combined administration of a DPP IV inhibitor or a salt thereof and at least one active ingredient selected from omega-3 oils, omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or triglycerides or a salt thereof, results not only in a beneficial, especially a synergistic, therapeutic effect, but also in additional benefits resulting from the combined treatment and further surprising beneficial effects compared to a monotherapy applying only one of the pharmaceutically active compounds used in the combinations disclosed herein.

It can be shown by established test models and especially those test models described herein that the combination of the DPP-IV inhibitor with at least one active ingredient selected from omega-3 oils, omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides results in a more effective prevention or preferably treatment of diseases specified in the following. In particular, it can be shown by established test models and especially those test models described herein that the combination of the present invention results in a more effective prevention or preferably treatment of diseases specified hereinafter.

If taken simultaneously, this results not only in a further enhanced beneficial, especially a synergistic, therapeutic effect, but also in additional benefits resulting from the simultaneous treatment such as a surprising prolongation of efficacy, a broader variety of therapeutic treatment and surprising beneficial effects conditions/disorders that might be treated by DPP-IV inhibition.

The term "potentiation" shall mean an increase of a corresponding pharmacological activity or therapeutical effect, respectively. Potentiation of one component of the combination according to the present invention by co-administration of another component according to the present invention means that an effect is being achieved that is greater than that achieved with one component alone.

The term "synergistic" shall mean that the drugs, when taken together, produce a total joint effect that is greater than the sum of the effects of each drug when taken alone.

Moreover, for a human patient, especially for elderly people, it is more convenient and easier to remember to take two tablets at the same time, e.g. before a meal, than staggered in time, i.e. according to a more complicated treatment schedule. More preferably, both active ingredients are administered as a fixed combination, i.e. as a single tablet, in all cases described herein. Taking a single tablet is even easier to handle than taking two tablets at the same time. Furthermore, the packaging can be accomplished with less effort.

The person skilled in the pertinent art is fully enabled to select a relevant and standard animal test model to prove the hereinbefore and hereinafter indicated therapeutic indications and beneficial effects.

The pharmaceutical activities as effected by administration of the combination of the active agents used according to the present invention can be demonstrated e.g. by using corresponding pharmacological models known in the pertinent art.

The insulin secretion enhancing properties of the combination according to the present invention may be determined by following the methodology as disclosed, for example, in the publication of T.lkenoue et al. Biol. Pharm. Bull. 29(4), 354-359 (1997).

The corresponding subject matter of these references is herewith incorporated by reference in this specification. Accordingly, the combination according to the present invention may be used, e.g., for the prevention, delay of progression or treatment of diseases and disorders that may be inhibited by DPP IV inhibition.

Thus in a further aspect the present invention concerns the use of a combination comprising a DPP IV inhibitor or a pharmaceutically acceptable salt thereof, and i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or ii) omega-3 oils, or in any case a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention, delay of progression or treatment of diseases and disorders that may be inhibited by DPP IV inhibition.

In a further aspect the present invention concerns the use of a DPP IV inhibitor or a pharmaceutically acceptable salt thereof, in combination with i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

The invention furthermore relates to a method for the prevention of, delay of progression of, treatment of diseases and disorders that may be inhibited by DPP IV inhibition, comprising administering to a warm-blooded animal, including man, in need thereof a jointly effective amount of a combination of a DPP IV inhibitor or a pharmaceutically acceptable salt thereof with; i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

The invention furthermore relates to a pharmaceutical composition for the prevention of, delay of progression of, treatment of a disease or condition selected from diseases and disorders that may be inhibited by DPP IV inhibition, comprising a combination of a DPP IV inhibitor or a pharmaceutically acceptable salt thereof with; i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

Combination, composition, use or method of treatment as described above, for the prevention, or delay of progression of disorders or conditions related/associated to diabetes (preferably type 2 diabetes).

Combination, composition, use or method of treatment as described above, for the prevention, or delay of progression of type 2 diabetes, in a patient suffering from IGM, preferably IGT.

Combination, composition, use or method of treatment as described above, for the prevention, reduction or delay in onset of cardiovascular diseases or conditions associated with diabetes (preferably type 2 diabetes), IGM or IGT, preferably selected from the group consisting of increased microvascular complications; increased cardiovascular morbidity; excess cerebrovascular diseases; increased cardiovascular mortality and sudden death, in a patient suffering from type 2 diabetes, IGM or IGT.

Most preferably, the disease or condition is selected from impaired glucose metabolism (IGM), conditions of impaired glucose tolerance, conditions of impaired fasting plasma glucose, diabetes particularly type 2 diabetes mellitus, diseases or conditions associated with diabetes, cardiovascular diseases or conditions associated with diabetes or IGM or IGT, diseases, disorders or conditions related/associated to diabetes, obesity, cognitive disorders and memory and learning ability problems, cognitive impairment associated with diabetes, impaired cognitive function associated with Alzheimer's disease, macular degeneration, dementias, Alzheimer's disease, altered gastrointestinal motility, sensitivity and/or secretion disorder(s), inflammatory bowel disease, arteriosclerosis, hyperlipidemia, hypertriglyceridemia, dyslipidemia, and conditions associated with hyperlipidemia.

Most preferably, the disease or condition is selected from conditions of impaired glucose tolerance, type 2 diabetes mellitus, cardiovascular diseases or conditions associated with diabetes or IGM or IGT, diseases, disorders or conditions related/associated to diabetes, obesity, cognitive disorders and memory and learning ability problems, cognitive impairment associated with diabetes, impaired cognitive function associated with Alzheimer's disease, Alzheimer's disease, macular degeneration, dementias, inflammatory bowel disease, arteriosclerosis, hyperlipidemia, hypertriglyceridemia or dyslipidemia.

A "disease or condition which may be inhibited by a DPP-IV inhibitor" as defined in this application comprises, but is not limited to insulin resistance, impaired glucose metabolism, conditions of impaired glucose tolerance (IGT), conditions of impaired fasting plasma glucose, diabetes particularly type 2 diabetes mellitus, obesity, diabetic retinopathy, macular degeneration, cataracts, diabetic nephropathy, glomerulosclerosis, diabetic neuropathy, erectile dysfunction, premenstrual syndrome, coronary heart disease, hypertension, angina pectoris, myocardial infarction, stroke, vascular restenosis, skin and connective tissue disorders, foot ulcerations and ulcerative colitis, endothelial dysfunction and impaired vascular compliance, diseases or conditions associated with diabetes or IGM or IGT, cardiovascular diseases or conditions associated with diabetes or IGM or IGT, diseases, disorders or conditions related/associated to diabetes, cardiovascular diseases or damages, neurodegenerative disorders, cognitive disorders and memory and learning ability problems. The neurodegenerative disorder is selected from Parkinson's disease, schizophrenia, dementia, senile dementia, mild cognitive impairment, Alzheimer related dementia, Huntington's chorea, tardive dyskinesia, hyperkinesias, mania, Morbus Parkinson, steel- Richard syndrome, Down's syndrome, myasthenia gravis, nerve and brain trauma, vascular amyloidosis, cerebral haemorrhage with amyloidosis, brain inflammation, Friedrich's ataxia, acute confusion disorders, acute confusion disorders in which apoptotic necrocytosis plays a part, amyotrophic lateral sclerosis, glaucoma, and Alzheimer's disease. The cognitive disorder is selected from cognitive deficits associated with schizophrenia, age-induced memory impairment, cognitive deficits associated with psychosis, cognitive impairment associated with diabetes, cognitive deficits associated with post-stroke, memory defects associated with hypoxia, cognitive and attention deficits associated with senile dementia, attention-deficit disorders, memory problems associated with mild cognitive impairment, impaired cognitive function associated with dementias, impaired cognitive function associated with Alzheimer's disease, impaired cognitive function associated with Parkinson's disease, impaired cognitive function associated with vascular dementia, cognitive problems associated with brain tumors, Pick's disease, cognitive deficits due to autism, cognitive deficits post electroconvulsive therapy, cognitive deficits associated with traumatic brain injury, amnesic disorders, deliriums, dementias, altered gastrointestinal motility, sensitivity and/or secretion disorder(s), dyslipidemia, hyperlipidemia, and conditions associated with hyperlipidemia.

The term "cardiovascular diseases or damages" as used herein includes cardiac hypertrophy, cardiac remodeling after myocardial infarction, pulmonary congestion and cardiac fibrosis in dilated or in hypertrophic cardiomyopathy, cardiomyopathy such as dilated cardiomyopathy or hypertrophic cardiomyopathy or diabetic cardiomyopathy, left or right ventricular hypertrophy, diabetic myopathy, stroke prevention in congestive heart failure, hypertrophic medial thickening in arteries and/or in large vessels, mesenteric vasculature hypertrophy, or artherosclerosis. Preferably the "cardiovascular diseases or conditions associated with diabetes (preferably type 2 diabetes), IGM or IGT" are preferably selected from the group consisting of increased microvascular complications; increased cardiovascular morbidity; excess cerebrovascular diseases; increased cardiovascular mortality and sudden death.

The diseases, disorders or conditions related/associated to diabetes, particularly type 2 diabetes mellitus, includes but are not limited to diabetic nephropathy, diabetic retinopathy and diabetic neuropathy, macular degeneration, coronary heart disease, myocardial infarction, diabetic cardiomyopathy, myocardial cell death, coronary artery diseases, peripheral arterial disease, stroke, limb ischemia, vascular restenosis, foot ulcerations, endothelial dysfunction and/or atherosclerosis.

The term "dementia" as used herein includes Alzheimer type dementia, Parkinson type dementia, Huntington type dementia, Pick's type dementia, Creutzfeldt-Jakob type dementia, senile dementia, pre-senile dementia, idiopathic-related dementia, trauma-related dementia, stroke-related dementia, cranial bleed- related dementia, vascular dementia, and includes acute, chronic or recurring forms.

Alzheimer's disease, is the most common form of dementia. This neurological disorder attacks the brain and results in cognitive problems, such as memory loss, impaired thinking, difficulty performing familiar tasks, disorientation to time and place, poor or decreased judgment, problems with language, changes in mood or behaviour and personality. Advancing age is the single greatest risk factor for Alzheimer's, a disease that strikes 10 percent of individuals by the time they reach age 65 and up to 50 percent by age 85. Part of this increased risk seems to occur because brain cells become increasingly vulnerable to stress as they age.

The term "Conditions associated with hyperlipidemia" include atherosclerosis, angina pectoris, carotid artery disease, cerebral arteriosclerosis, xanthoma, CHD, heart attaks, ischemic stroke, restenosis after angioplasty, peripheral vascular disease, intermittent claudication, myocardial infarction (e.g. reduction in necrosis), dyslipidemia, post-prandial lipemia.

The term "hyperlipidemia" refers to the presence of an abnormally elevated level of lipids in the blood. Hyperlipidemia can appear in at least three forms: (1) hypercholesterolemia, i.e., an elevated cholesterol level; (2) hypertriglyceridemia, i.e., an elevated triglyceride level; and (3) combined hyperlipidemia, i.e., a combination of hypercholesterolemia and hypertriglyceridemia. This term also refers to elevated levels of one or more lipoproteins, e.g., elevated levels of Lp(a), LDL and/or VLDL.

The term "cholesterol" refers to a steroid alcohol that is an essential component of cell membranes and myelin sheaths and, as used herein, incorporates its common usage. Cholesterol also serves as a precursor for steroid hormones and bile acids. The term "triglyceride(s)" (TGs), as used herein, incorporates its common usage. TGs consist of three fatty acid molecules esterified to a glycerol molecule and serve to store fatty acids which are used by muscle cells for energy production or are taken up and stored in adipose tissue.

Because cholesterol and TGs are water insoluble, they must be packaged in special molecular complexes known as "lipoproteins" in order to be transported in the plasma. Lipoproteins can accumulate in the plasma due to overproduction and/or deficient removal. There are at least five distinct lipoproteins differing in size, composition, density and function. In the cells of the small of the intestine, dietary lipids are packaged into large lipoprotein complexes called "chylomicrons", which have a high TG and low cholesterol content. In the liver, TG and cholesterol esters are packaged and released into plasma as TG-rich lipoprotein called VLDL, whose primary function is the endogenous transport of TGs made in the liver or released by adipose tissue. Through enzymatic action, VLDL can be either reduced and taken up by the liver, or transformed into IDL. IDL, is in turn, either taken up by the liver, or is further modified to form the LDL. LDL is either taken up and broken down by the liver, or is taken up by extrahepatic tissue. HDL helps remove cholesterol from peripheral tissues in a process called reverse cholesterol transport. Exemplary primary hyperlipidemia include, but are not limited to, the following: 1) Familial hyperchylomicronemia, a rare genetic disorder which causes a deficiency in an enzyme, LP lipase, that breaks down fat molecules. The LP lipase deficiency can cause the accumulation of large quantities of fat or lipoproteins in the blood;

2) Familial hypercholesterolemia, a relatively common genetic disorder caused where the underlying defect is a series of mutations in the LDL receptor gene that result in malfunctioning LDL receptors and/or absence of the LDL receptors. This brings about ineffective clearance of LDL by the LDL receptors resulting in elevated LDL and total cholesterol levels in the plasma;

3) Familial combined hyperlipidemia, also known as multiple lipoprotein-type hyperlipidemia; an inherited disorder where patients and their affected first-degree relatives can at various times manifest high cholesterol and high triglycerides. Levels of HDL cholesterol are often moderately decreased;

4) Familial defective apolipoprotein B-100 is a relatively common autosomal dominant genetic abnormality. The defect is caused by a single nucleotide mutation that produces a substitution of glutamine for arginine which can cause reduced affinity of LDL particles for the LDL receptor. Consequently, this can cause high plasma LDL and total cholesterol levels;

5) Familial dysbetaliproteinemia, also referred to as Type III hyperlipoproteinemia, is an uncommon inherited disorder resulting in moderate to severe elevations of serum TG and cholesterol levels with abnormal apolipoprotein E function. HDL levels are usually normal; and

6) Familial hypertriglyceridemia, is a common inherited disorder in which the concentration of plasma VLDL is elevated. This can cause mild to moderately elevated triglyceride levels (and usually not cholesterol levels) and can often be associated with low plasma HDL levels. Risk factors in exemplary secondary hyperlipidemia include, but are not limited to, the following: (1) disease risk factors, such as a history of Type 1 diabetes, Type 2 diabetes, Cushing's syndrome, hypothyroidism, cholestasis and certain types of renal failure; (2) drug risk factors, which include, birth control pills; hormones, such as estrogen and corticosteroids; certain diuretics; and various β-blockers; (3) dietary risk factors include dietary fat intake per total calories greater than 40%; saturated fat intake per total calories greater than 10%; cholesterol intake greater than 300 mg per day; habitual and excessive alcohol use; bulimia, anorexia nervosa, and obesity. The term "altered gastrointestinal motility, sensitivity and/or secretion disorder(s)" as used herein includes one or more of the symptoms and conditions which affect the gastrointestinal tract from the mouth to the anus, which include, but are not limited to, heartburn, bloating, postoperative ileus, abdominal pain and discomfort, early satiety, epigastric pain, nausea, vomiting, burbulence, regurgitation, intestinal pseudoobstruction, anal incontinence, GERD, IBS, inflammatory bowel disease (IBD), dyspepsia, chronic constipation or diarrhea, diabetic gastropathy, gastroparesis, e.g. diabetic gastroparesis, ulcerative colitis, Crohn's disease, ulcers and the visceral pain associated therewith. Preferably IBD, IBS, GERD and Crohn's disease.

Preferably, a "disease or condition which may be inhibited by a DPP-IV inhibitor" is selected from impaired glucose metabolism (IGM), conditions of impaired glucose tolerance, conditions of impaired fasting plasma glucose, diabetes particularly type 2 diabetes mellitus, diseases or conditions associated with diabetes, cardiovascular diseases or conditions associated with diabetes or IGM or IGT, diseases, disorders or conditions related/associated to diabetes, obesity, cognitive disorders and memory and learning ability problems, cognitive impairment associated with diabetes, impaired cognitive function associated with Alzheimer's disease, dementias, Alzheimer's disease, macular degeneration, altered gastrointestinal motility, sensitivity and/or secretion disorder(s), inflammatory bowel disease, arteriosclerosis, hyperlipidemia, hypertriglyceridemia, dyslipidemia, and conditions associated with hyperlipidemia.

Preferably, a "disease or condition which may be inhibited by a DPP-IV inhibitor" is selected from conditions of impaired glucose tolerance, type 2 diabetes mellitus, cardiovascular diseases or conditions associated with diabetes or IGM or IGT, diseases, disorders or conditions related/associated to diabetes, obesity, cognitive disorders and memory and learning ability problems, cognitive impairment associated with diabetes, impaired cognitive function associated with Alzheimer's disease, Alzheimer's disease, dementias, macular degeneration, inflammatory bowel disease, arteriosclerosis, hyperlipidemia, hypertriglyceridemia or dyslipidemia.

The term "curative" as used herein means efficacy in treating ongoing diseases, disorder or conditions.

The term "prophylactic or prevention" means the prevention of the onset or recurrence of diseases, disorders or conditions to be treated. The term "delay of progression" as used herein means administration of the combination to patients being in a pre-stage or in an early phase of the disease to be treated, in which patients for example a pre-form of the corresponding disease is diagnosed or which patients are in a condition, e.g. during a medical treatment or a condition resulting from an accident, under which it is likely that a corresponding disease will develop.

The term "combined pharmaceutical preparation" as that term is used herein means that the active ingredients, i.e. omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega- 3 mono-, di-, or tri-glycerides, or omega-3 oils, and a DPP-IV inhibitor preferably LAF237, are both administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body, preferably at the same time. As an example, a non-fixed combination would be two capsules each containing one active ingredient where the purpose is to have the patient achieve treatment with both active ingredients together in the body.

The term "fatty acid oxidation" relates to the conversion of fatty acids (e.g., oleate) into ketone bodies.

The term "modulate" means to induce any change including increasing or decreasing, (e.g., a modulator of fatty acid oxidation increases or decreases the rate of fatty oxidation, a modulator of a receptor includes both agonists and antagonists of the receptor).

The term "weight loss" refers to loss of a portion of total body weight.

The term "treat" or "treatment" encompasses the complete range of therapeutically positive effects associated with pharmaceutical medication including reduction of, alleviation of and relief from the symptoms or illness which affect the organism.

Preferably, the jointly therapeutically effective amounts of the active agents according to the combination of the present invention can be administered simultaneously or sequentially in any order, e.g. separately (combined pharmaceutical preparation) or in a fixed combination.

Under certain circumstances, drugs with different mechanisms of action may be combined. However, just considering any combination of drugs having different modes of action but acting in the similar field does not necessarily lead to combinations with advantageous effects. AII the more surprising is the experimental finding that the combined administration of a DPP-IV inhibitor according to the present invention, or, in each case, a pharmaceutically acceptable form thereof, results not only in a beneficial, especially a potentiating or a synergistic, therapeutic effect. Independent thereof, additional benefits resulting from combined treatment can be achieved such as a surprising prolongation of efficacy, a broader variety of therapeutic treatment and surprising beneficial effects on diseases and conditions associated with diabetes (e.g. less triglycerides, less glycemia, less cardiovascular diseases, less gain of weight or less cardiovascular side effects).

Further benefits are that lower doses of the individual drugs to be combined according to the present invention can be used to reduce the dosage, for example, that the dosages need not only often be smaller but are also applied less frequently, or can be used in order to diminish the incidence of side effects. This is in accordance with the desires and requirements of the patients to be treated.

In a further embodiment, the present invention concerns a combination, composition, use or method of treatment, as described herein, wherein the patients to be treated is suffering from type 2 diabetes, obesity, hyperlipidemia or hypertriglyceridemia.

In a further embodiment, the present invention concerns a combination, composition, use or method of treatment, as described herein, wherein the patient to be treated has at least a triglyceride level of 500 mg/dl.

For example, it has turned out that the combination according to the present invention provides benefit especially in the treatment of diabetic patients, e.g. reducing the risk of negative cardiovascular events, reducing risk of side effects, controlling increase of weight, reduction of hyperlipidemia, hypertriglyceridemia especially in diabetic patients or in patients suffering from an altered gastrointestinal motility, sensitivity and/or secretion disorders).

In view of reduced dose of the DPP-IV inhibitor or, omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or omega-3 oils, used according to the present invention, there is a considerable safety profile of the combination making it suitable for first line therapy.

The pharmaceutical composition according to the present invention as described herein before and hereinafter may be used for simultaneous use or sequential use in any order, for separate use or as a fixed combination. Method or use as described above, wherein the DPP-IV inhibitor and at least one active ingredient selected from omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or omega-3 oils are administered in the form of a combination of the present invention such as a fixed combination or combined preparation or kit of part.

According the invention, when the DPP-IV inhibitors, and at least one active ingredient selected from omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono- , di-, or tri-glycerides, or omega-3 oils are administered together, such administration can be sequential in time or simultaneous with, the simultaneous method being generally preferred. For sequential administration, the DPP-IV inhibitor, and and at least one active ingredient selected from omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono- , di-, or tri-glycerides, or omega-3 oils can be administered in any order. It is generally preferred that such administration be oral. It is especially preferred that the administration be oral and simultaneous. However, if the subject being treated is unable to swallow, or oral absorption is otherwise impaired or undesirable, parenteral or transdermal administration will be appropriate. When the DPP-IV inhibitor, and at least one active ingredient selected from omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or triglycerides, or omega-3 oils are administered sequentially, the administration of each can be by the same method or by different methods.

A further aspect of the present invention is a kit for the prevention of, delay of progression of, treatment of a disease or condition according to the present invention comprising

(a) an amount of a DPP IV inhibitor or a pharmaceutically acceptable salt thereof in a first unit dosage form;

(b) an amount of at least one active ingredient selected from omega-3 fatty acids, omega- 3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or omega-3 oils or, in each case, where appropriate, a pharmaceutically acceptable salt thereof in a second etc. unit dosage form; and

(c) a container for containing said first, second etc. unit forms.

In a variation thereof, the present invention likewise relates to a "kit-of-parts", for example, in the sense that the components to be combined according to the present invention can be dosed independently or by use of different fixed combinations with distinguished amounts of the components, i.e. simultaneously or at different time points. The parts of the kit of parts can then e.g. be administered simultaneously or chronologically staggered, that is at different time points and with equal or different time intervals for any part of the kit of parts. Preferably, the time intervals are chosen such that the effect on the treated disease or condition in the combined use of the parts is larger than the effect that would be obtained by use of only any one of the components.

The present invention thus also relates to a kit of parts comprising

(b) an amount of at least one active ingredient selected from omega-3 fatty acids, omega- 3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or omega-3 oils or, in each case, where appropriate, a pharmaceutically acceptable salt thereof, in the form of two or three or more separate units of the components (a) to (b), especially for the prevention of, delay of progression of, treatment of a disease or condition according to the present invention.

The invention furthermore relates to a commercial package comprising the combination according to the present invention together with instructions for simultaneous, separate or sequential use.

In a preferred embodiment, the (commercial) product is a commercial package comprising as active ingredients the combination according to the present invention (in the form of two or three or more separate units of the components (a) or (b)), together with instructions for its simultaneous, separate or sequential use, or any combination thereof, in the delay of progression or treatment of the diseases as mentioned herein.

All the preferences mentioned herein apply to the combination, composition, use, method of treatment, "kit of parts" and commercial package of the invention.

These pharmaceutical preparations are for enteral, such as oral, and also rectal or parenteral, administration to homeotherms, with the preparations comprising the pharmacological active compound either alone or together with customary pharmaceutical auxiliary substances. For example, the pharmaceutical preparations consist of from about 0.1 % to 90 %, preferably of from about 1 % to about 80 %, of the active compound. Pharmaceutical preparations for enteral or parenteral, and also for ocular, administration are, for example, in unit dose forms, such as coated tablets, tablets, capsules or suppositories and also ampoules. These are prepared in a manner that is known per se, for example using conventional mixing, granulation, coating, solubulizing or lyophilizing processes. Thus, pharmaceutical preparations for oral use can be obtained by combining the active compound(s) with solid excipients, if desired granulating a mixture which has been obtained, and, if required or necessary, processing the mixture or granulate into tablets or coated tablet cores after having added suitable auxiliary substances.

The dosage of the active compound can depend on a variety of factors, such as mode of administration, homeothermic species, age and/or individual condition.

Preferred dosages for the active ingredients of the pharmaceutical combination according to the present invention are therapeutically effective dosages, especially those which are commercially available.

Normally, in the case of oral administration, an approximate daily dose of from about 1 mg to about 360 mg is to be estimated e.g. for a patient of approximately 75 kg in weight.

The pharmaceutical preparation will be supplied in the form of suitable dosage unit form, for example, a capsule or tablet, and comprising an amount, being together with the further component(s) jointly effective, e.g. 100 mg or 50 mg of vildagliptin.

The pharmaceutical composition according to the present invention as described hereinbefore may be used for simultaneous use or sequential use in any order, for separate use or as a fixed combination.

Thus according to a further embodiment, a DPP-IV inhibitor, is administered with and at least one active ingredient selected from omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or omega-3 oils, preferably in the form of a fixed pharmaceutical composition comprising a pharmaceutically acceptable carrier, vehicle or diluent. Accordingly, a DPP-IV inhibitor of this invention, can be administered with at least one active ingredient selected from omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or omega-3 oils as a fixed combination, in any conventional oral, parenteral or transdermal dosage form.

The doses of DPP-IV inhibitor of formula (I) to be administered to warm-blooded animals, for example human beings, of, for example, approximately 70 kg body weight, especially the doses effective in the inhibition of the DPP-IV enzyme, are from approximately 3 mg to approximately 3 g, preferably from approximately 10 mg to approximately 1 g, for example approximately from 20 mg to 200 mg, per person per day, divided preferably into 1 to 4 single doses which may, for example, be of the same size. Usually, children receive about half of the adult dose. The dose necessary for each individual can be monitored, for example by measuring the serum concentration of the active ingredient, and adjusted to an optimum level. Single doses comprise, for example, 10, 40 or 100 mg per adult patient.

The dosage of vildagliptin is preferably between 10 and 150 mg daily, most preferably between 25 and 150 mg, 25 and 100 mg or 25 and 50 mg or 50-100 mg daily. Preferred examples of daily oral dosage are 25, 30, 35, 45, 50, 55, 60, 80, 100 or 150 mg. The application of the active ingredient may occur up to three times a day, preferably one or two times a day.

The preferred herein mentioned omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or omega-3 oils will be supplied in the form of suitable dosage unit form, for example, a capsule or tablet, and comprising a therapeutically effective amount, e.g. from about 100 to about 5000 mg, as already described herein and in the prior art. The application of the active ingredient may occur up to three times a day, preferably one or two times a day. The same preferred dosage are selected for the fixed combinations.

Daily omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or omega-3 oils dosages required in practicing the method of the present invention will vary depending upon, for example the mode of administration and the severity of the condition to be treated. An indicated daily dose is in the range of from about 100 to about 5000 mg, e.g. from 500 to 4000 mg or from 500 to 3600 mg, or e.g. from 500 to 2000 mg of active agent for oral use, conveniently administered once or in divided dosages.

Corresponding doses may be taken, for example, in the morning, at mid-day or in the evening.

In a preferred aspect, the invention concerns a "kit-of-parts", combination, composition, use or a method as described herein, comprising or wherein the daily dosage administration is; i) between 25 and 150 mg or between 50 and 100 mg of vildagliptin, and ii) between 500 and 5000 mg or between 500 and 4000 mg of at least one active ingredient selected from omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or omega-3 oils, or in any case, a pharmaceutically acceptable salt thereof.

In a preferred aspect, the invention concerns a "kit-of-parts", combination, composition, use or a method as described herein, comprising or wherein the daily dosage administration is; i) 25, 50, 100 or 150 mg of vildagliptin, and ii) 500, 1000, 2000 or 4000 mg of at least one active ingredient selected from omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or omega-3 oils, or in any case, a pharmaceutically acceptable salt thereof.

In a preferred aspect, the invention concerns a "kit-of-parts", combination, composition, use or a method as described herein, wherein the omega-3 fatty acid is selected from DHA , EPA or ALA .

Preferred are the herein described "kit-of-parts", combinations, compositions, uses or methods of treatment, wherein the DPP-IV inhibitor is preferably vildagliptin or a pharmaceutically accepted salt thereof and wherein the at least one second active agent i) is selected from the group consisting of EPA, DHA or ALA, preferably EPA and DHA ,or in any case a pharmaceutically accepted salt thereof.

Preferred are the herein described "kit-of-parts", combinations, compositions, uses or methods of treatment, wherein the DPP-IV inhibitor is preferably vildagliptin or a pharmaceutically accepted salt thereof and wherein the at least one second active agent i) is selected from a mixture of EPA and DHA or in any case a pharmaceutically accepted salt thereof.

Preferred are the herein described "kit-of-parts", combinations, compositions, uses or methods of treatment, wherein the DPP-IV inhibitor is preferably vildagliptin or a pharmaceutically accepted salt thereof and wherein the omega-3 oil ii) is selected from Epax®, Omegabrite®, Epanova®, E463808, E681010, Omacor®, OMEGA- 3/90, Incromega, Epadel®, Seacor®, Esapent®, Eskim®, Liposol®.

In another embodiment, the herein described "kit-of-parts", combinations, compositions, uses or methods of treatment, comprise about 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, or 1500 mg of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides or a mixture thereof.

In another embodiment, the herein described "kit-of-parts", combinations, compositions, uses or methods of treatment comprise a daily dosage of between 25 and 150 mg or between 50 and 100 mg of a DPP4 inhibitor, preferably vildagliptin, and a daily dosage of i) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg, or e.g. between 500 to 2000 mg of omega-3 oil, or ii) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg, or e.g. between 500 to 2000 mg of at least one active agent selected from the group consisting of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters and omega-3 mono-, di-, or tri-glycerides, or iii) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg, or e.g. between 500 to 2000 mg of at least one active agent selected from the group consisting of EPA, ALA and DHA preferably EPA and DHA, or iv) between 100 and 5000 mg, or between 100 to 4000 mg, or between, or e.g. between 500 to 2000 mg mg of at least two active agents which is a mixture of EPA and DHA, or v) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg, or e.g. between 500 to 2000 mg of at least one active agent selected from the group consisting of ethyl esters of EPA, ethyl esters of ALA and ethyl esters of DHA, or vi) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg, or e.g. between 500 to 2000 mg of at least one omega-3 oil is selected from Epax®, Omegabrite®, Epanova®, E463808, E681010, Omacor®, OMEGA- 3/90, Incromega, Epadel®, Seacor®, Esapent®, Eskim® and Liposol®, or vii) about 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, or 1500 mg of at least one active agent selected from the group consisting of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters and omega-3 mono-, di-, or tri-glycerides, or viii) about 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, or 1500 mg of at least one active agent selected from the group consisting of EPA, ALA and DHA, or ix) between 500 and 4000 mg, preferably 1000 and 4000 mg of Omacor® or in any case a pharmaceutically accepted salt thereof.

By the term e.g. "between 100 to 4000 mg of at least one second active agent selected from the group consisting of EPA, ALA or DHA or in any case a pharmaceutically accepted salt thereof the applicant means that the weight of EPA, DHA or ALA alone, or the weight of a mixture of any of them is between 500 to 3000 mg.

By the term "at least one second active agent" the applicant means 1 , 2, 3, 4 or more active agents, preferably 1 or 2.

Preferably, in case of free combinations, preferred are those dosages for launched products that have been approved and that have been marketed.

Especially preferred are low dose combinations.

To further illustrate the invention, but not by way of limitation, the following examples are provided.

Experimental part:

Example 1 : Bioassay Methods for Assessing the Effects of Compounds and Combination Therapies on Body Fat Reduction, Body Weight, and Lipid Metabolism.

The dose(s) administered to the animal are sufficient to determine if the compounds or combination therapy has a desired effect, for example, an appetite, body weight, body fat, and/or fatty acid oxidation over time. Such dose(s) can be determined according to the efficacy of the particular candidate compound(s) employed and the condition of the animal, as well as the body weight or surface area of the animal. The size of the dose(s) also will be determined by the existence, nature, and extent of any adverse side effects that accompany the administration of a candidate compound or combination; the LD50 of the candidate compound or combination; and the side-effects of the candidate compound or combination at various concentrations. Depending upon the compound or combination and the above factors, for instance, the initial test dosage(s) may range, for example, from 0.1-50 mg per kg, preferably 1-25 mg per kg, most preferably 1-20 mg per kg body weight for each of the compound or combination. The determination of dose response relationships is well known to one of ordinary skill in the art.

Test animals subjects can be, for example, obese or normal mammals (e.g., humans, primates, guinea pigs, rats, mice, or rabbits). Suitable rats include, but are not limited to, Zucker rats. Suitable mice include, but are not limited to, for example, ALS/LtJ, C3. SW-H- 2b/SnJ, (NON/LtJ x NZO/HIJ)F1, NZO/H1J, ALR/LtJ, NON/LtJ, KK.Cg- AALR/LU, NON/LtJ, KK.Cg-Ay/J, B6.HRS(BKS)-Cpefat/+, B6.129P2-Gcktm/Efr, B6.V-Lepob, BKS.Cg- m+/+Leprdb, and C57BL/6J with Diet Induced Obesity.

Example 2: Assessing Effects on Body Fat Reduction.

Effects on body fat can be identified in vivo using animal bioassay techniques well known to those of ordinary skill in the art. Body fat reduction is typically determined by direct measurements of the change in body fat or by loss of body weight. Body fat and/or body weight of the animals is determined before, during, and after the administration of the candidate compounds or combinations. Test compounds (DPP-IV inhibitors (vildagliptin) and omega-3 oils (e.g. Omacor) or combinations thereof and appropriate vehicle or caloric controls can be administered by any of a number of routes (e.g., the oral route, a parenteral route) to experimental subjects and the weight of the subjects can be monitored over the course of therapy. The experimental subjects can be humans as well as surrogate test animals (e. g., rats, mice).

Changes in body fat are measured by any means known in the art such as, for example, fat fold measurements with calipers, bioelectrical impedance, hydrostatic weighing, or dual x-ray absorbiometry. Preferably animals demonstrate at least 2%, 5%, 8%, or 10% loss of body fat. Changes in body weight can be measured by any means known in the art such as, for example, on a portable scale, on a digital scale, on a balance scale, on a floor scale, or a table scale. Preferably animals demonstrate at least 2%, 5%, 10%, or 15% loss of body weight. Body weight reduction is measured before administration of the candidate compound or combination and at regular intervals during and after treatment. Preferably, body weight is measured every 5 days, more preferably every 4 days, even more preferably every 3 days, yet more preferably every 2 days, most preferably every day.

For instance, the effect of the candidate compounds and combinations on total body fat can be determined by taking direct measurements of the rat's body fat using skin fold calipers. Skin on the subjects' backs, abdomen, chest, front and rear legs can be pinched with calipers to obtain measurements before administration of the test compound and at daily or longer intervals (e.g., every 48 hours) during and after administration of candidate compounds and combinations. Differences in measurements in one or more of the "pinched" sites reflect the change in the rat's total body fat. The animal may selected from any test species, including but not limited to, mammals, the mouse, a rat, a guinea pig, or a rabbit. The animal may also be an ob/ob mouse, a db/db mouse, or a Zucker rat or other animal model for a weight-associated disease. Clinical studies in humans may also be conducted. In humans, body density measurements or estimates of percent body fat may also be used to assess body fat reduction.

Example 3: Assessing Effects on Lipid Metabolism.

The candidate compounds and combinations i.e. DPP-IV inhibitors (vildagliptin) and omega- 3 oils (e.g. Omacor) or combinations of such compounds can also be assayed for their effect on fatty acid metabolism. The effect of the candidate compounds and combinations on fatty acid metabolism can be measured by measurements of fatty acid oxidation in primary cultures of liver cells as taught for instance in U.S. Patent application Ser. No. 10/112,509 filed on Mar. 27, 2002 and assigned to the same assignee as the present application and incorporated by reference.

Changes in fatty acid metabolism can be measured, for instance, by looking at fatty acid oxidation in cells from major fat burning tissues such as, for example, liver (Beynen, et al., Diabetes, 28:828 (1979)), muscle (Chiasson Lab. Anat. of Rat (1980)), heart (Flink, et al., J. Biol. Chem., 267: 9917 (1992)), and adipocytes (Rodbell, J. Biol. Chem., 239: 375 (1964)), Cells may be from primary cultures or from cell lines. Cells may be prepared for primary cultures by any means known in the art including, for example, enzymatic digestion and dissection. Suitable cell lines are known to those in the art. Suitable hepatocyte lines are, for example, Fao, MH1C1 , H-4-II-E, H4TG, H4-II-E-C3, McA-RH7777, McA-RH8994, N1-S1 Fudr, N1-S1 , ARL-6, Hepa 1-6, Hepa-1c1c7, BpRd , tao BpRd , NCTC clone 1469, PLC/PRF/5, Hep 3B2.1- 7 [Hep 3B], Hep G2 [HepG2J, SK-HEP-1 , WCH-17. Suitable skeletal muscle cell lines are, for example, L6, L8, C8, NOR-10, BLO-11 , BC3H1 , G-7, G-8, C2C12, P19, Solδ, SJRH30 [RMS 13], QM7. Suitable cardiac cell lines are, for example, H9c2(2-1), P19, CCD-32Lu, CCD-32Sk, Girardi, FBHE. Suitable adipocyte lines are, for example, NCTC clone 929 [derivative of Strain L; L-929; L cell], NCTC 2071 , L-M, L-M(TK-) [LMTK-; LM(tk-)], A9 (APRT and HPRT negative derivative of Strain L), NCTC clone 2472, NCTC clone 2555, 3T3-L1 , J26, J27-neo, J27-B7, MTKP 97-12 pMp97B [TKMp97-12], L- NGC-5HT2, Ltk-11 , L-alpha-1b, L-alpha-2A, L-alpha-2C, B82.

The rate of fatty acid oxidation may be measured by 14C- oleate oxidation to ketone bodies (Guzman and Geelen Biochem. J. 287:487 (1982)) and/or 14C-oleate oxidation to CO₂ (Fruebis, PNAS, 98:2005 (2001); Blazquez, et al., J. Neurochem, 71 : 1597 (1998) ). Lypolysis may be measured by fatty acid or glycerol release by using appropriate labeled precursors or spectrophotometric assays (Serradeil-Le GaI₁ FEBS Lett, 475: 150 (2000)). For analysis of 14C-oleate oxidation to ketone bodies, freshly isolated cells or cultured cell lines can be incubated with 14C-oleic acid for an appropriate time, such as, for example, 30, 60, 90, 120, or 180 minutes. The amount of 14C radioactivity in the incubation medium can be measured to determine their rate of oleate oxidation. Oleate oxidation can be expressed as nmol oleate produced in x minutes per g cells. For analysis of lypolysis/glycerol release, freshly isolated cells or cultured cells lines can be washed then incubated for an appropriate time. The amount of glycerol released into the incubation media can provide an index for lypolysis.

Example 4: Evaluation of the effects the LAF237 and Amacor on human lipid profiles. Sixty (60) patients comprising male and non-fertile female patients aged at least 30 years with a diagnosis of Type 2 diabetes mellitus of at least three months duration, who have been treated with diet alone for at least one month prior to study entry are selected. The study is broken down into two periods and 3 group of patients (LAF237 alone, Omacor alone, and the combination LAF237 and Omacor). Period 1 is the four weeks prior to the beginning of the study, with period 2 being four weeks and being the actual study period when patients are treated with LAF237 alone, Omacor alone, or the combination LAF237 and Omacor. Accordingly, study entry is Week -4 and the endpoint is after the fourth week of Period 2.

Patients are randomized in a ratio of 1 :1 :1 as follows: 50 mg once a day (OD) of LAF237 alone, 4 g once a day of Omacor alone, and the combination LAF237 and Omacor 50mg/4g OD. The patients receive active ingredients 30 minutes before breakfast. There are 5 test days in the study. Patients attended as outpatients for fasting blood sampling at Week -4 (study entry), Week -2 and Week 2 and as inpatients for 24 hours on Week 0 (= baseline) and Week 4 (= endpoint). On the two inpatient test days, the total caloric intake of breakfast, lunch and dinner are standardized and standard test meals are administered in place of breakfast and dinner. Triglycerides, total cholesterol and lipid fractions (LDL, VLDL and HDL) are measured during 24 hours following the breakfast standard meal. On the three outpatient test days, patients fast for at least 7 hours (i.e., no food or drinks (except water) after midnight on the day before the scheduled visit) and attend between 07.00 and 10.00 h and do not take the morning dose of active ingredient. On the two inpatient test days, patients fast for at least 7 hours, i.e., no food or drinks (except water) after midnight on the day before the scheduled visit, and attend the clinic at 07.00 h. On each of the two test days, the total caloric intake during 24 hours is standardized and standard test meals is administered for breakfast (about 08.00 h) and dinner (about 18.00 h). Lunch is taken at approximately 13.00 h. On Day 1 , no active ingredient is administered but at Week 4, patients take active ingredient(s) as normal, 30 minutes before the standard breakfast. Triglycerides, total cholesterol and lipid fractions (LDL, VLDL and HDL) are evaluated. Triglycerides, total cholesterol and HDL are measured and LDL and VLDL calculated according to the method of Friedewald et al., "Estimation of the Concentration of Low-Density Lipoprotein Cholesterol Without the Use of the Preparative Centrifuge", Clin. Chem., Vol. 18, No. 6, pp. 499-502 (1972).

This study can show a much higher lowering of levels of triglyceride, total cholesterol, LDL and VLDL of the combination compared to placebo or monotherapy, preferably the combination shows more than an additive effect.

Example 5: Effects of Omacor and LAF237 on gastric and colonic sensitivity to distension and on the muscular tone of the gut using barostatic distension.

1. 1. Gastric sensitivity and tone

Groups of Wistar rats weighing 200-250 g are used. For surgery, the animals are preme- dicated with 0.3 ml of acepromazine (0.5 mg/kg) injected intraperitoneally (ip) and anesthetized with 0.3 ml of ketamine injected intraperitoneally.

Animals are positioned in dorsal decubitus and following a xypho-ombilical laparotomy, the stomach is fitted with a permanent balloon connected to a tube introduced in the upper part of the rumen at 1 cm of the gastro-esophageal junction on the great curvature. After closure of the abdomen, rats are positioned in ventral decubitus and one group of 3 stainless steel electrodes (1 m in length - 270 μm in diameter) is implanted into the neck muscles using a technique described in Ruckebusch and Fioramonti, Gastroenterol. 68:1500-1508,1975. The free ends of electrodes and the catheter of the balloon are exteriorized on the back of the neck and protected by a glass tube attached to the skin.

Gastric distension at constant pressure is performed with an electronic barostat (Hachet et al., Gastroenterol Clin Biol, 1993, 17, 347-351). Balloons (5.0-5.5 cm in length) are made with cistern free condoms and sutured to a polyethylene tube (1.0 and 1.8 mm inner and outer diameter respectively, 80 cm in length). The end of the tube is drilled for an easier emptying of the balloon. Ten days after surgery, electromyographic recordings are performed with an electroencephalograph machine (Reega VIII, Alvar, Paris, France) at a paper speed of 2.4 cm/min. A short time constant of amplification is used to record selectively spike burst (0.03 s). The electromyographic activity is summed every 20 s by an integrator circuit and automatically plotted on a computer.

Under noxious gastric distension, the rat stretches its body and rises up the head and/or turns the head on the left and right sides to observe his flank. The neck muscles are contracted and an electromyographic signal is recorded. In addition, the barostat is connected to a potentiometric recorder for the permanent recording of intragastric pressure. The animals are separated into groups.

After a 30 min period of control recording, the animals receive one of the following regimens: 1) placebo, 2) Amacor, 3) LAF237, 4) Amacor + LAF237.

The protocol of gastric distension is started 30 min later.

Electromyographic activity of the neck muscles (EANM) is correlated with changes of posture and is proportional to pain induced by gastric distension. Values integrated every 20 s are summed up for consecutive 10 min. For each stage of distension, neck activity is determined with the following formula :

(EANM at a determined pressure ) - (EANM in basal conditions) *100

EANM in basal conditions

The pain threshold is determined as an increase > 100 % of the electrical activity of the neck muscles.

Gastric volume is determined on the potentiometric recorder as the maximal volume obtained for each stage of distension. Pain threshold and gastric volume are given as mean

± SEM and values compared using Student's T test for unpaired values.

The pharmaceutical combination of Amacor and LAF237 can significantly decrease the gastric pain associated with gastric distension and increases in gastric tone as compared to placebo and any of the compounds administered alone.

2. Colorectal sensitivity and tone

The influence of Amacor and LAF237 on rectal or colonic tone and pain is done using barostat distension procedures by applying increasing pressure in a stair-case manner for consecutive periods of 5min.; the volume is measured for each pressure giving an evaluation of the changes in tone.

Wistar rats weighing 220-250 g and housed individually are used. The animals are premedicated with 0.5 mg/kg of acepromazine injected intraperitoneally (IP) and anesthetized by intramuscular administration of 100 mg/kg of ketamine. They are prepared for electromyographic recordings using the technique described in Ruckebusch and Fioramonti, 1975. Pairs of nichrome wire electrodes (60 cm in length and 80 μm in diameter) are implanted in the striated muscle of the abdomen, 2 cm laterally from the white line. The free ends of electrodes are exteriorized on the back of the neck and protected by a plastic tube attached to the skin.

Electromyographic recordings (time constant : 0.03 sec) started 8 days after surgery. Bipolar recordings of myoelectric activity are performed with an electroencephalographic recorder during one hour starting 30 min before rectal distension.

In order to prevent recording artefacts due to movements during distension, rats are acclimated, 3 days before distension, to stay in tunnel of polypropylene in which distension and EMG recordings are performed. A balloon consisting of a condom (4cm) is introduced into the rectum at 5 cm from the anus and fixed at the base of the tail. The balloon, connected to a barostat, is increasingly inflated with air at pressures of 15, 30, 45 and 60mmHg. each pressure being applied during 5min.

Groups of rats are submitted respectively to the barostatic distension protocol. Ten minutes before they are injected IP with 1) placebo, 2) Amacor, 3) LAF237, 4) Amacor + LAF237. Statistical analysis of the number of abdominal spike bursts occurring during each 5 min period are performed by Student's "f test fair paired values comparisons after two way ANOVA. P<0.05 is considered statistically significant. Colorectal volumes are given as mean ± SEM and values compared using Student's "f test for unpaired values. The pharmaceutical combination of Amacor plus LAF237 can significantly decreases the rectal and colonic pain associated with rectal distension and increases colorectal tone as compared to placebo and any of the compounds administered alone.

Example 6: Bioassay Methods for Assessing the Effects of Compounds, and Combination Therapies on Alzheimer's disease, Parkinson's disease, cognitive disorders, memory and learning ability problems.

The pharmacological activity of the compounds i.e. DPP-IV inhibitors (LAF237) and omega-3 fatty acid e.g. Amacor and combinations of such compounds according to the invention in improving cognitive function may, for example, be assessed using tests known to a person skilled in the art such as standardized psychometric tests (e.g. Wechsler Memory Scale, the Wechsler Adult Intelligence Scale, Raven's Standard Progressive Matrices, Schaie- Thurstone Adult Mental Abilities Test), neuropsychological tests (e.g. Luria-Nebraska), metacognitive self-evaluations (e.g. Metamemory Questionnaire), visual-spatial screening tests (e.g. Poppelreuter's Figures, Clock Recognition, Honeycomb Drawing and Cancellation), cognitive screening tests (e.g. Folstein's Mini Mental State Test) and reaction time tests. Such standardized tests as listed above are described in Ruoppila.l. and Suutama.T. (1997) Scand. J. Soc. Med. Suppl. 53, 44-65 and serve as examples, said reference is incorporated in its entirety herein. The term "cognitive function" includes the functions assessed by any such test.

A clinical protocol to show the positive effect of a the combination of the invention for treating the Alzheimer's disease development is described in the patent application WO 2004/082706 on pages 31-37, which is incorporated herein by reference.

A further useful in vivo protocol which can be used to show that the claimed combinations can improving cognitive function is described in the European Patent No.1310258 (examples 5-8).

The examples 2-4 described in the patent application WO2005009349 describe a further protocols to assess the activity of the compounds and combinations of the present invention to treat or prevent, X syndrome , Alzheimer's disease and Parkinson's' disease which is incorporated herein by reference.

Other standard tests for cognitive performance e.g. the Alzheimer's Disease Assessment Scale (ADAS- cog) are described by Doraiswamy (Neurology. 1997 Jun;48(6):1511-7) and in the patents US20040024043 and US 6369046. The ADAS-cog is a multi-item instrument for measuring cognitive performance which include elements of memory, orientation, retention, reasoning, language and praxis. US20040024043 describes also an in vivo test model in rodents in example 5 and a clinical Study Design in example 9. Another clinical Study Design is described by US 6369046 (example 1) which are incorporated herein by reference.

A further useful in vivo protocol which can be used to show that DPP-IV inhibitors can improving cognitive function is described in the European Patent No.1310258 (examples 5- 8), which is incorporated herein by reference.

Example 7: Parkinson disease The effects of the herein described combinations i.e. comprising a DPP-IV inhibitors such as vildagliptin (LAF237) and an Omega-3 oil such as Amacor in a model of Parkinson disease are investigated in mice. Male C57/BL6 mice are injected once daily for 7 days with MPTP (30 mg/kg, i.p.). Vildagliptin and/or Amacor are administered once or twice daily for 14 days. On day 28, striata are removed, homogenized in perchloric acid, and centrifuged. The supernatant is removed and analyzed for dopamine and other monoamines such as serotonin by reverse-phase HPLC and electrochemical detection. Anti-Parkinson activity of the combination Vildagliptin and Amacor is assessed in comparison to the mono-therapy with either Vildagliptin or Amacor alone.

Example 8: Treating, Preventing or Delaying cognitive impairment associated with e.g. diabetes, Alzheimer's disease or with Parkinson's disease.

One 50 mg tablet of vildagliptin, or 4 g of Amacor or a combination of 50 mg of vildagliptin and 4 g of Amacor are administered daily with water to subjects in need of such treatment. The cognitive status of the subject is monitored periodically using the MMSE (Mini Mental State Examination (MMSE) with norms adjusted for age and education (Folstein et al., J. Psych. Res., 12 (1975), 196-198, Anthony et al., Psychological Med.¹ 12 (1982), 397-408; Cockrell et al., Psychopharmacology, 24 (1988), 689-692; Crum et al., J. Am. Med. Assoc'n. 18 (1993), 2386-2391) or similar tool.

Example 9: Treatment for Preventing or Delaying the Onset of Alzheimer's Disease e.g. in a Subject Exhibiting Mild Cognitive Impairment + Dementia.

A subject having e.g. mild cognitive impairment, is identified using the MMSE or similar diagnostic tool e.g. ADAS- cog.

A 50 mg tablet of vildagliptin or 2 g of Amacor or a combination of 50 mg of vildagliptin and

2 g of Amacor are administered daily with water to said subject.

The subject is monitored periodically using the MMSE or similar tool e.g. ADAS- cog, to assess the evolution of Alzheimer's Disease and the cognitive impairment. The subject is also monitored for clinical symptoms of dementia.

Example 10: Treatment for Preventing or Delaying the Onset of Alzheimer's Disease in a Subject Exhibiting Mild Cognitive Impairment Subjects having mild cognitive impairment is identified using the MMSE or similar diagnostic tool.

Subjects are separated into 3 groups. A 50 mg tablet of vildagliptin or 2 g of Amacor or a combination of 50 mg of vildagliptin and 2 g of Amacor are administered daily with water to said subject.

The cognitive status of the subjects is monitored periodically using the MMSE or similar tool, and the subject is monitored for clinical symptoms of dementia.

Example 3 - Treating, Preventing or Delaying cognitive impairment associated with diabetes Subjects are separated into 3 groups. A 50 mg tablet of vildagliptin or 2 g of Amacor or a combination of 50 mg of vildagliptin and 2 g of Amacor are administered daily with water to said subject.

The cognitive status of the subjects is monitored periodically using the MMSE or similar tool.

Example 11 : Learning deficits

The ameliorative effect of vildagliptin or the combined use of vildagliptin with Omega-3 oils e.g. Amacor, on learning deficits is investigated in aged rats. The following methods describe a set of experiments using the vildagliptin as monotherapy or a combination of vildagliptin with Omega-3 oils e.g. Amacor.

Methods Male (3 to 27 months old) rats of transgenic strain are used. The aged rats are divided into the following four groups (the active ingredient dosage can be adapted by the person skilled in the art).

1) Control group: Repeated administration of placebo pill.

2) Vildagliptin group: Repeated oral administration of vildagliptin 3 mg/kg.

3) Cholinesterase inhibitor group: Repeated oral administration of Amacor 60 mg/kg.

4) Combination group: Repeated oral administration of vildagliptin 3 mg/kg and Amacor 60 mg/kg.

In the combination group, vildagliptin is administered 30 minutes after administration of Amacor.

Passive avoidance learning test is started on day 14 of treatment, and Morris water maze learning test on day 20 of treatment. On each day of experiment, vildagliptin and/or Amacor are administered 30 minutes and I hour, respectively, before initiation of the trial.

1. Passive Avoidance Learning: The passive avoidance learning test is performed using a chamber consisting of light and dark compartments. Young rats (pill, 10 animals) and aged rats (control group, 10 animals; vildagliptin group, 10 animals, Amacor group, 10 animals; combination group, 10 animals) are individually placed in the light compartment and 10 seconds later, the sliding door is opened. After a mouse moves to the dark compartment, the mouse is kept there for about 10 seconds with the door closed. One to two hours after the habituation trial, acquisition trial is performed.

In the acquisition trial, after a mouse moved to the dark compartment, a foot shock (0.4 mA, 3 seconds) is given through the grid floor. Retention trials are performed 24 hours after acquisition trials.

In each trial, the latency from opening of the slide door until the animal moved to the dark compartment (step-through latency) is measured.

2. Morris Water Maze Learning Same animals used in the passive avoidance test are subjected for the water maze task. However, some rats can not swim well in the water tank, thus they are excluded in the water maze task. The water maze learning test is performed on young rats (saline, 10 animals) and aged rats (control group, 9 animals; vildagliptin group, 9 animals: Amacor group, 8 animals; combination group, 8 animals).

In pretraining which is performed for swimming training and motivation for escaping from water, four trials are performed using a water bath, 80 cm in diameter, in a condition that the platform is visible. From the following day, using a water bath, 120 cm in diameter, learning trials, one session (four trials) per day, are performed with the platform being placed below the water.

1. Passive Avoidance Learning

The control group will show a significant decrease in the avoidance time as compared with the young group. The test can accordingly assess that the vildagliptin group and the done Amacor group show significant improvement of the learning deficit in aged rats. The test can accordingly assess an improvement of the cognitive status of the treated subject. This test can also indicate that combination of vildagliptin and Amacor improves the learning deficit in aged rats, and this effect is greater than that seen when either drug is used alone. It can furthermore show that the combination has improved results or advantages than that seen when either drug is used alone.

2. Water Maze Learning

In the water maze task, the control group will show a significant prolongation of latency to find platform submerged in the water compared with the young rats. The test can accordingly assess that vildagliptin group and the Amacor group show a significant improvement in water maze learning deficit. The test can accordingly assess an improvement of the cognitive status of the treated subject.

This test can also indicate that combination of vildagliptin and Amacor improves water maze learning deficit in aged rats, and this effect is greater than that seen when either drug is used alone. It can furthermore show that the combination has improved results or advantages than that seen when either drug is used alone.

Other tests may be performed using animal models e.g. of dementia such as some of those described and reviewed in the following references: Higgins L.S., Vol. Med Today 1999, 5(6):274-6; Borchelt D.R. et al., Brain Pathol. 1998, 8(4):735-57 and Guenette S.Y. et al., Neurobiol. Aging 1999, 20(2):201 -11.

Although the present invention has been described in considerable detail with reference to certain preferred versions thereof, other versions are possible without departing from the spirit and scope of the preferred versions contained herein. All references and Patents (U.S. and others) referred to herein are hereby incorporated by reference in their entirety as if set forth herein in full.

Example 12: Strokes:

WO0110867 describes protocols which can be applied to assess the advantages of vildagliptin for the treatment or prevention of strokes.

Example 13: Dyslipidemia

Diet-induced obese mice are used for the study at 21-23 weeks of age. On the first day of the study, animals are fasted at 7:30 a.m. Body weight measurement and basal blood sample collection are conducted at 10:30 a.m. Animals were assigned into two groups (n = 10/group) with the plasma glucose values and body weights matched between the two groups. The animals are dosed orally with vehicle (water) or compound at 30 mg/kg at a dose volume of 5 ml/kg. Daily dose of vehicle or the compound is administered at the same time each day for a total of 15 days. Daily body weight and food intake measurements are taken during the study. Fat and lean mass analyses are performed at twice during the 15 day period using the EchoMRI Whole Body Composition Analyzer. Scans are performed using the appropriate size holders provided by the manufacturer. On the last day of the study (day 15), mice are fasted at 7:30 a.m. and dosed with vehicle or compound at 10:30 a.m. Tail blood samples are taken at 12:30 p.m. Animals are then euthanized with carbon dioxide. Terminal blood samples are collected via cardiac puncture for blood chemistry analysis.

Blood collection and analyses

Blood samples are taken during the study via tail bleeding. Plasma glucose concentrations are determined using a glucose meter (Ascensia Elite, Bayer Corp., Mishawaka, IN). Blood samples were collected in tubes (Microvette CB300, Aktiengesellschaft & Co., Numbrecht, Germany) which contain lithium heparin to prevent blood clotting. Prior to each blood sample collection, 1 μl of 1 :10 diluted protease inhibitor cocktail (Sigma, St. Louis, MO) is added to the sample tubes. After blood sample collection, the tubes are kept on ice before being centrifuged. The plasma portion of the blood samples is obtained by centrifugation at 10,000 x g for 10 min at 4 ⁰C and then stored at -80 ⁰C. Plasma insulin levels are determined by Luminex assays using Mouse Endocrine Lincop/ex kit (Linco Research, Inc., St. Charles, MO). Plasma triglyceride, fatty acid and total cholesterol levels are determined using a fluorescent assay based on Amplex Red kit (Molecular Probes, Eugene, OR). Animals treated with drug show a lowering in plasma triglyceride, free fatty acid and cholesterol levels as compared to the control animals. Blood chemistry analysis is performed using an automated dry chemistry system (SPOTCHEM EZ Analyzer, Heska, Fort Collins, CO).

Example 14: Neurodegenerative diseases

Positive effect on Neurodegenerative diseases is assessed by the following experimental

Animals

Female B6CBAF1/J mice are transplanted with ovaries from female

B6CBATg(HDexon1)62Gpb/1J mice (R6/2, Mangiarini et al., 1996) are obtained from

Jackson Laboratories (Bar Harbor, Maine), and bred to B6CBAF1/J (The Jackson Laboratory, Maine) male mice in house. In a sample of 45 mice (males and females) from our colony, the repeat length measures 119-130. Mice (R6/2 transgenic (TG), n = 27, male = 14, female = 13), derived from 7 litters, are tested from 21 days until 70 days of age and euthanized at 84 days (see Fig. 1 for details of testing time-points). Mice are housed at a temperature (21-23⁰C) and humidity (30-70%) controlled room with food and water available ad lib, using a reverse light-dark cycle (10 am lights off, 10 pm lights on). Genotypes are determined from DNA from tail snips taken at 3 weeks of age. DNA is isolated using a kit from Qiagen (Valencia, CA). CAG repeats are sized by PCR using FAM-labeled primers (5'- ATGAAGGCCTTCGAGTCCCTCAAGTCCTTC-3') and (5'-

GGCGGCTGAGGAAGCTGAGGA-3') in AM buffer (67 mM Tris-HCI [pH 8.8], 16.6 mM NH4SO4, 2.0mMMgCI2 0.17 mg/ml BSA, 1OmM 2-mercaptoethanol), 10% DMSO, 200 mM dNTPs, 8 ng/μl primers with 0.5 U/ml Taq polymerase. Cycling conditions were 90" @ 94⁰C, 25 X (30" @ 94⁰C, 30" @ 65⁰C, 90" @ 72⁰C), 10' @ 72⁰C. The PCR products are sized using an ABI sequencer and the Genescan and Genotyper software packages. The size of the CAG repeat is 85 bp less than the size of the PCR product. All experiments are performed in accordance with and approved by the Institutional Animal Care and Use Committee at NIBRI.

Compounds dosing

Mice are weighed weekly from 21 days until 84 days of age. At 21 days of age, the animals are divided into 4 groups and dosed orally with either vehicle (water) or compounds (LAF237 or Omacor) at 30 mg/kg and 60 mg/kg respectively at a dose volume of 5 ml/kg. Daily dose of vehicle or the compounds or the combination (LAF237 + Omacor) is administered at the same time each day for a total of 63 days. Testing for behavioral phenotype is performed at the end of the treatment period. The age of death for each mouse is recorded.

Behavioral tests Running wheel

Mice are placed individually in cages equipped with a running wheel (23 cm diameter, Mini Mitter Company Inc., Bend OR). Each rotation of the wheel is detected by a magnet and recorded by VitalView Data Acquisition Software V 4.0 (Mini Mitter Company Inc. as above), in 3 min bins. Running wheel cages are housed in cabinets (8 cages/cabinet) to minimize light and sound disturbance (lights off 10 am, lights on 10 pm). Running activity is recorded continuously for 6-8 days (majority housed for 7-8 days WT, n = 12, R6/2 n = 21). Wheel running activity during light and dark phases is calculated using ActiView V 1.2 (Mini Mitter Company Inc. as above). Measurements are calculated from daily running activities or from the middle section of each exposure to the running wheels (3rd, 4th, and 5th full day in running wheels at 4.5-5.5 weeks, or 8.5-9.5 weeks). Measurements include: (1) the maximum number of rotations per time-bin (3 min) during the 3rd, 4th, and 5th day, (2) the average activity per time-bin during the light and the dark phases, derived from each successive day in the running wheels, (3) the average light and the average dark activity per time-bin, derived from activities averaged over the 3rd, 4th and 5th day in the running wheels, (4) the number of breaks taken during the night phases of the 3rd, 4th, and 5th night, (5) the total number of rotations run over the 3rd, 4th, and 5th full day in the running wheels.

Climbing cage

Climbing behavior is video-recorded during the light phase, to finish at least 1 h before the beginning of the dark phase. Mice are habituated to the testing room and then placed underneath a wire cylinder (diameter, 10.5 cm, height, 15.5 cm, open at bottom end) for 5 min. The latency to climb, the total time spent climbing, the number of instances of climbing and rearing are counted from videos by a blinded observer. Climbing is defined as when all four paws of the mouse are off the floor of the testing bench and on the wall of the climbing cage.

Open field

Spontaneous activity in the open field is monitored using an automated system that detected movement with infrared beam breaks (Truscan system for mice, Coulbourn Instruments, Allentown, PA). Beam break information is collected using Truscan V 1.012.00 software. Mice are placed individually in the center of the open field (25.5 cm square) and monitored for 15 min. Testing takes place halfway through the dark cycle. An observer is present at all times and the open field is illuminated with an angle-poise lamp equipped with a 25-W red bulb. Distance covered in the horizontal plane and rearing is analyzed in 3 - 5 min time bins.

Rotarod

The rotarod apparatus (Ugo Basile, Varese, Italy) is used to measure motor coordination and balance. The axle is covered with smooth rubber to prevent the mice clinging to the axle. Testing is carried out approximately half way through the dark phase, using a red light (25 W) for illumination. Following 15-20 min habituation to the testing room, mice are given 3 trials (at least 10 min between trials) on an accelerating protocol (4-40 rpm in 10 min) similar to other published protocols. The latency to fall is measured. If a mouse falls off the rotarod in less than 20 s, it is placed back on immediately (up to 3 times). Mice are tested over 4 days at baseline (4 weeks of age) and over 3 days at 8 weeks of age. Mean latency to fall per mouse at baseline and 8 weeks of age is calculated and used to generate group means.

Grip strength

A spring weigh scales (Fisher Scientific, Tustin, CA) with an attached trapeze is hung from a wall mount. Mice were allowed to grasp the trapeze with their forepaws, while the observer pulled down gently on the mouse' tail. The weight pulled minus body weight is used for analysis. Mice are given 5 trials, from which the 3 best scores are used for analysis.

Example 15: Type 2 diabetes/Metabolic disease

Diet-induced obese mice are used for the study at 21-23 weeks of age. On the first day of the study, animals are fasted at 7:30 a.m. Body weight measurement and basal blood sample collection are conducted at 10:30 a.m. Plasma glucose values are then determined. Animals were assigned into 4 groups (n = 10/group) with the plasma glucose values and body weights matched between the 4 groups. At 12 pm, the animals are dosed orally with vehicle (water) or the compounds individually (LAF27 or Omacor) at 30 mg/kg or 60 m/kg respectively at a dose volume of 5 ml/kg or the corresponding combination. At 1 :00 p.m. a blood sample (at 0 min) is taken followed by an oral glucose tolerance test (OGTT) at 1 g/kg (20% glucose in water) at a dose volume of 5 ml/kg. Blood samples are collected at 30, 60 and 120 min following the glucose administration. The animals are refed after the OGTT. The animals are administered a daily dose of vehicle or the compounds or combinations 12:00 p.m. each day for a total of 15 days. Daily body weight and food intake measurements are performed during the study. Two additional OGTTs are performed during the study on the days 7 and 14, following the protocol described above for the OGTT on day 1. Animals treated with drugs (mono or combination therapy) show an improvement in glucose tolerance as compared to the control animals, as measured by the area under the curve during an OGTT. Animals treated with the combination show an improved result. The magnitude of improvement in the OGTT increases in a time-dependent manner from day 7 to day 14. On the last day of the study (day 15), mice are fasted at 7:30 a.m. and dosed with vehicle or compounds or combination at 10:30 a.m. Tail blood samples are taken at 12:30 p.m. Animals are then euthanized with carbon dioxide. Terminal blood samples are collected via cardiac puncture for blood chemistry analysis. Blood collection and analyses

Blood samples are taken during the study via tail bleeding. Plasma glucose concentrations are determined using a glucose meter (Ascensia Elite, Bayer Corp., Mishawaka, IN). Blood samples were collected in tubes (Microvette CB300, Aktiengesellschaft & Co., Numbrecht, Germany) which contain lithium heparin to prevent blood clotting. Prior to each blood sample collection, 1 μl of 1 :10 diluted protease inhibitor cocktail (Sigma, St. Louis, MO) is added to the sample tubes. After blood sample collection, the tubes are kept on ice before being centrifuged. The plasma portion of the blood samples is obtained by centrifugation at 10,000 x g for 10 min at 4 ⁰C and then stored at -80 ⁰C. Plasma insulin and glucagons levels are determined by Luminex assays using Mouse Endocrine Lincop/ex kit (Linco Research, Inc., St. Charles, MO). Animals treated with drugs or combination show a lowering in plasma insulin levels as compared to the control animals. Plasma triglyceride, fatty acid and total cholesterol levels are determined using a fluorescent assay based on Amplex Red kit (Molecular Probes, Eugene, OR). Blood chemistry analysis is performed using an automated dry chemistry system (SPOTCHEM EZ Analyzer, Heska, Fort Collins, CO).

Example 16: IGM patients: Treatment, prevention of type 2 diabetes and cardiovascular diseases.

Favorable effects can be verified that confirm that the claimed combinations can restore early phase insulin secretion and reduce post-prandial glucose levels in subjects with IGM, or can prevent or delay the onset of Type 2 diabetes and cardiovascular diseases in subjects with IGM. A multi-center, double-blind, parallel group, randomized study can be conducted in subjects with IGM in order to evaluate the incidence of confirmed hypoglycemia and the effects on prandial glucose associated with the administration of 50 mg of LAF237, 2 g of Omacor, or the combination of LAF237 and Omacor or placebo before each main meal during 8 weeks of treatment. Subjects are selected on the basis of a 2-hour plasma glucose value after a 75 g oral glucose tolerance test (OGTT) and patients essentially meeting the following additional inclusion criteria are included in the study:

- two-hour glycemia post-OGTT between 7.8 to 11.1 mmol/L (one OGTT to be performed during the year before entering the study, the second to be performed within two weeks prior entering the study);

- FPG < 7 mmol/L;

- patients are to have a body mass index (BMI) between 20-32 kg/m2;

- patients are to maintain prior diet during the full course of study;

- males, non-fertile females, females of child-bearing potential using a medically approved birth control method are included;

- the use of other antidiabetics during the trial is not permitted.

Corresponding dosages of e.g. LAF or Omacor or the corresponding combination are administered with a large glass of water 2 (BID), 3 (TID) or 4 (QID) times daily depending on the number of main meals (breakfast, lunch, snack, dinner). The first dose is to be given with the first main meal (standardized meal i.e. 55% carbohydrates, 25% fat and 20% protein). Visits are scheduled to be performed at weeks 0, 2, 4 and 8 and the patients are to be fasted for at least 7 hours. All blood samples for laboratory evaluations are drawn between 07.00 and 10.00 a.m. HbAIc is to be measured at baseline and after 8 weeks of treatment (fasting glucose and fructosamine). Samples of blood are to be drawn at 10, 20, 20, 60, 120, and 180 minutes after drug administration (time 0) and the glucose and insulin levels to be measured. At weeks 0 and 8 visits, patients complete a standard meal challenge containing approximately 500 kcal and measurements of insulin and glucose will be performed.

The findings from analyses of all obtained data in such a study can reveal that 2 hour prandial glucose levels, HBAIc and fructosamine levels can be reduced, that early phase insulin secretion can be restored, and that such a combination can prevent or delay the progression to type 2 diabetes mellitus. With longer treatment and follow-up, the preventive and reduction effect on conditions and diseases associated with IGM e.g. cardiovascular diseases can be evaluated i.e. prevention or delay of progression to overt diabetes mellitus type 2; or prevention, reduction or delay in onset of a cardiovascular condition or disease associated with IGT preferably selected from the group consisting of increased microvascular complications; increased cardiovascular morbidity; excess cerebrovascular diseases; increased cardiovascular mortality and sudden death

This type of study in individuals with IGM and particularly IFG and IGT differs from those in diabetics since the subjects have normal FPG and are non-diabetics or pre-diabetics.

In all the above examples, the effect of the combination is preferably more than additive. Determining the Combination Therapy Dosages.

Preferred dosages of the Omega-3 oils and DPP-IV inhibitor to be used in a combination therapy can be determined experimentally by first conducting separate dose response studies for the Omega-3 oils and DPP-IV inhibitor to be used. Methods of performing such dose response studies in a test species or the species of the intended subject (e.g., a human) are well known to one of ordinary skill in the art. The endpoint of the study is preferably selected according to the effect or endpoint of interest (e.g., weight loss, body fat reduction, changes in lipid metabolism, changed food seeking behavior) Or the dose response of the underlying mechanism of action (e.g., receptor activation or antagonism). Alternatively, the established dose response relationships may be used if an agent is already well-characterized as to dose response. Preferred bioassay methods include those described above and those presented in the Examples. AII patents and literature references cited in this specification are hereby incorporated by reference in their entirety. In case of inconsistencies, the present description, including the definitions and interpretations, will prevail.

The invention has been described above by reference to preferred embodiments but, as those skilled in the art will appreciate, many additions, omissions and modifications are possible all within the scope of the claims below.

Although the present invention has been described in considerable detail with reference to certain preferred versions thereof, other versions are possible without departing from the spirit and scope of the preferred versions contained herein. All references and Patents (U.S. and others) referred to herein are hereby incorporated by reference in their entirety as if set forth in full herein.

Claims

What is claimed is

1) A combination, such as a combined preparation or pharmaceutical composition, respectively, comprising a DPP IV inhibitor or a pharmaceutically acceptable salt thereof, and

1) at least one compound selected from the group consisting of omega-3 fatty acids, omega- 3 esters, omega-3 alkyl esters or omega-3 mono-, di-, or tri-glycerides, or ii) omega-3 oils, or in any case a pharmaceutically acceptable salt thereof.

2) A combined preparation or pharmaceutical composition, respectively, comprising a DPP IV inhibitor or a pharmaceutically acceptable salt thereof, and i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

3) Use of a DPP IV inhibitor or a pharmaceutically acceptable salt thereof, in combination with; i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

4) A method for the prevention of, delay of progression of, treatment of diseases and disorders that may be inhibited by DPP IV inhibition, comprising administering to a warm-blooded animal, including man, in need thereof a jointly effective amount of a combination of a DPP IV inhibitor or a pharmaceutically acceptable salt thereof with; i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

5) A pharmaceutical composition for the prevention of, delay of progression of, treatment of a disease or condition selected from diseases and disorders that may be inhibited by DPP IV inhibition, comprising a combination of a DPP IV inhibitor or a pharmaceutically acceptable salt thereof with; i) at least one compound selected from the group consisting of omega-3 fatty acids, omega-

6) Composition, use or method of treatment according to any of claims 3 to 5, for the prevention, or delay of progression, of disorders or conditions related/associated to diabetes.

7) Composition, use or method of treatment according to any of claims 3 to 5, for the prevention, or delay of progression of type 2 diabetes, in a patient suffering from IGM, preferably IGT.

8) Composition, use or method of treatment according to any of claims 3 to 5, for the prevention, reduction or delay in onset of cardiovascular diseases or conditions associated with diabetes (preferably type 2 diabetes), IGM or IGT, preferably selected from the group consisting of increased microvascular complications; increased cardiovascular morbidity; excess cerebrovascular diseases; increased cardiovascular mortality and sudden death, in a patient suffering from type 2 diabetes, IGM or IGT.

9) Composition, use or method of treatment according to any of claims 3 to 5, wherein the disease or condition which may be inhibited by a DPP-IV inhibitor is selected from impaired glucose metabolism (IGM), conditions of impaired glucose tolerance, conditions of impaired fasting plasma glucose, diabetes particularly type 2 diabetes mellitus, diseases or conditions associated with diabetes, cardiovascular diseases or conditions associated with diabetes or IGM or IGT, diseases, disorders or conditions related/associated to diabetes, obesity, cognitive disorders and memory and learning ability problems, cognitive impairment associated with diabetes, impaired cognitive function associated with Alzheimer's disease, dementias, Alzheimer's disease, macular degeneration, altered gastrointestinal motility, sensitivity and/or secretion disorder(s), inflammatory bowel disease, arteriosclerosis, hyperlipidemia, hypertriglyceridemia, dyslipidemia, and conditions associated with hyperlipidemia. 10) A combination, composition, use or method of treatment, according to any of the previous claims, wherein the at least one second active agent i), is selected from the group consisting of EPA, DHA or ALA, preferably EPA and DHA ,or in any case a pharmaceutically accepted salt thereof.

11) A combination, composition, use or method of treatment, according to any of the previous claims, wherein the at least one second active agent i) is selected from a mixture of EPA and DHA or in any case a pharmaceutically accepted salt thereof.

12) A combination, composition, use or method of treatment, wherein the omega-3 oil ii), is selected from Epax®, Omegabrite®, Epanova®, E463808, E681010, Omacor®, OMEGA- 3/90, Incromega, Epadel®, Seacor®, Esapent®, Eskim®, Liposol®.

13) A combination according to claim 1 , composition according to claims 2 or 5, use according to any of claims 3, 6, 7, 8 or 9, or a method of treatment according to any of claims 4, 6, 7, 8 or 9, which comprises a daily dosage of between 25 and 150 mg or between 50 and 100 mg of a DPP4 inhibitor, preferably vildagliptin, and a daily dosage of i) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg of omega-3 oil, or ii) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg of at least one active agent selected from the group consisting of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters and omega-3 mono-, di-, or triglycerides, or iii) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg of at least one active agent selected from the group consisting of EPA, ALA and DHA preferably EPA and DHA, or iv) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg of at least two active agents which is a mixture of EPA and DHA, or v) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg of at least one active agent selected from the group consisting of ethyl esters of EPA, ethyl esters of ALA and ethyl esters of DHA, or vi) between 100 and 5000 mg, or between 100 to 4000 mg, or between 500 to 4000 mg of at least one omega-3 oil is selected from Epax®, Omegabrite®, Epanova®, E463808, E681010, Omacor®, OMEGA- 3/90, Incromega, Epadel®, Seacor®, Esapent®, Eskim®¹ and Liposol®, or vii) about 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, or 1500 mg of at least one active agent selected from the group consisting of omega-3 fatty acids, omega-3 esters, omega-3 alkyl esters and omega-3 mono-, di-, or tri-glycerides, or viii) about 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, or 1500 mg of at least one active agent selected from the group consisting of EPA, ALA and DHA, or ix) between 500 and 4000 mg, preferably 1000 and 4000 mg of Omacor® or in any case a pharmaceutically accepted salt thereof.

14) A combination, composition, use or method of treatment, according to any of the previous claims, wherein the DPP-IV inhibitor is selected from (S)-1 -{2-[5-cyanopyridin- 2yl)amino]ethyl-aminoacetyl)-2-cyano- pyrrolidine, vildagliptin, MK-0431 (Sitagliptin), (2S)-1- { (2-(5-Methyl-2-phenyl-oxazol-4-yl)-ethylamino)-acetyl}-pyrrolidine-2-carbonitrile, or (2S)-1-{ (1 , 1 -Dimethyl-3-(4-pyridin-3-yl-imidazol-1 -yl)-propylamino)-acetyl}-pyrrolidine-2 -carbonitrile, (S)-1-( (2S,3S, 11bS)-2-Amino-9, 10-dimethoxy-1 , 3,4,6,7, 11 b-hexahydro-2H-pyrido (2,1 -a) isoquinolin-3-yl)-4-fluoromethyl-pyrrolidin-2-one, or (S, S₁S₁S)-I -(2-Amino-9, 10-dimethoxy- 1 ,3,4,6,7,1 11b-hexahydro-2H-pyrido (2,1 -a) isoquinolin-3-yl)-4-methyl-pyrrolidin-2-one, GSK23A, saxagliptin, 3-(aminomethyl)-2-isobuthyl-1-oxo-4-phenyl-1 ,2-dihydro-6- isoquinolinecarboxamide and 2-{[3-(aminomethyl)-2-isobuthyl-4-phenyl-1-oxo-1 ,2-dihydro-6- isoquinolyl]oxy}acetamide, or in each case, a pharmaceutically acceptable salt thereof.

15) A combination, composition, use or method of treatment, according to any of the previous claims, wherein the DPP-IV inhibitor is vildagliptin or a pharmaceutically acceptable salt thereof.

16) A combination, composition, use or method of treatment, according to any of the previous claims, wherein vildagliptin or a pharmaceutically acceptable salt thereof, is administered in an amount between 25 and 150 mg or between 50 and 100 mg daily.

17) A combination, composition, use or method of treatment, according to any of the previous claims, wherein the patients to be treated is suffering from type 2 diabetes, obesity, hyperlipidemia, or hypertriglyceridemia. 18) A combination, composition, use or method of treatment, according to any of the previous claims, wherein the patient to be treated has at least a triglyceride level of 500 mg/dl.