Combination therapy for the treatment of diabetes and conditions related thereto and for the treatment of conditions ameliorated by increasing a blood glp-1 level

Abstract

The present invention concerns combination of an amount of a GPR119 agonist with an amount of a dipeptidyl peptidase IV (DPP-IV) inhibitor such that the combination provides an effect in lowering a blood glucose level or in increasing a blood GLP-1 level in a subject over that provided by the amount of the GPR119 agonist or the amount of the DPP-IV inhibitor alone and the use of such a combination for treating or preventing diabetes and conditions related thereto or conditions ameliorated by increasing a blood GLP-1 level. The present invention also relates to the use of a G protein-coupled receptor to screen for GLP-1 secretagogues.

1 .- 47 . (canceled) 
     
     
         48 . A method for identifying a GLP-1 secretagogue, comprising:
 (a) contacting a test compound with a host cell or with a membrane of a host cell that expresses a G protein-coupled receptor (GPCR), wherein the GPCR comprises an amino acid sequence selected from the group consisting of:
 (i) amino acids 2-335 of SEQ ID NO:2; 
 (ii) the amino acid sequence of a GPCR encoded by a polynucleotide comprising a nucleotide sequence, wherein the nucleotide sequence is capable of hybridizing under stringent conditions to a complement of the sequence set forth in SEQ ID NO:1; and 
 (iii) a biologically active fragment of the GPCR of (i) or (ii); 
   (b) determining the ability of the test compound to stimulate functionality of the GPCR;   (c) contacting the test compound that stimulates functionality of the GPCR in step (b) with a mammalian enteroendocrine cell in vitro; and   (d) determining whether the test compound stimulates GLP-1 secretion from the mammalian enteroendocrine cell,   wherein the ability of the test compound to stimulate GLP-1 secretion from the mammalian enteroendocrine cell identifies the test compound as the GLP-1 secretagogue.   
     
     
         49 . The method of  claim 48 , wherein the test compound is selected from the group consisting of a small molecule, a polypeptide, an antibody, and an antigen-binding fragment of the antibody. 
     
     
         50 . The method of  claim 48 , wherein the host cell comprises an expression vector, the expression vector comprising a polynucleotide encoding the GPCR. 
     
     
         51 . The method of  claim 48 , wherein the determining the ability of the test compound to stimulate functionality of the GPCR is through the measurement of the level of a second messenger selected from the group consisting of cyclic AMP (cAMP), cyclic GMP (cGMP), inositol 1,4,5-triphosphate (IP3), diacylglycerol (DAG), MAP kinase activity, MAPK/ERK kinase kinase-1 (MEKK1) activity, and Ca2+. 
     
     
         52 . The method of  claim 51 , wherein the second messenger is cAMP. 
     
     
         53 . The method of  claim 48 , wherein the determining the ability of the test compound to stimulate functionality of the GPCR is through the use of a Melanophore assay, or through the measurement of GTPγS binding to a membrane comprising the GPCR. 
     
     
         54 . The method of  claim 48 , wherein the GPCR is recombinant. 
     
     
         55 . The method of  claim 48 , wherein the host cell is an enteroendocrine cell of a GLUTag-Fro cell line. 
     
     
         56 . The method of  claim 48 , wherein the GPCR comprises amino acids 2-335 of SEQ ID NO:2. 
     
     
         57 . The method of  claim 48 , wherein the GPCR comprises the amino acid sequence of SEQ ID NO:2. 
     
     
         58 . A composition comprising a G protein-coupled receptor 119 (GPR119) agonist and a Dipeptidyl Peptidase IV (DPP-IV inhibitor), wherein the DPP-IV inhibitor is not identical to 1-[2-[5-cyanopyridin-2-yl)amino]ethylamino]acetyl-2-cyano-(S)-pyrrolidine (NVP-DPP728). 
     
     
         59 . A method of identifying a GLP-1 secretagogue, comprising:
 (a) contacting a GPR119 agonist with a mammalian enteroendocrine cell in vitro; and   (b) determining whether the GPR119 agonist stimulates GLP-1 secretion from the mammalian enteroendocrine cell;   wherein the ability of the GPR119 agonist to stimulate GLP-1 secretion from the mammalian enteroendocrine cell identifies the agonist as a GLP-1 secretagogue.   
     
     
         60 . A method of identifying a GLP-1 secretagogue, comprising:
 (a) administering a GPR119 agonist to a mammal; and   (b) determining a blood GLP-1 level in a biological sample obtained from the mammal,   wherein the ability of the GPR119 agonist to increase a blood GLP-1 level in the biological sample obtained from the mammal identifies the agonist as a GLP-1 secretagogue.   
     
     
         61 . A method of identifying a GLP-1 secretagogue, comprising determining a blood GLP-1 level in a biological sample obtained from a mammal, said mammal having been administered with a GPR119 agonist, wherein the ability of the GPR119 agonist to increase a blood GLP-1 level in the biological sample obtained from the mammal identifies the agonist as a GLP-1 secretagogue. 
     
     
         62 . A method for identifying a GLP-1 secretagogue, comprising:
 (a) contacting a GPCR with a ligand to the GPCR in the presence of a test compound, wherein the GPCR comprises an amino acid sequence selected from the group consisting of:
 (i) amino acids 2-335 of SEQ ID NO:2; 
 (ii) the amino acid sequence of a GPCR encoded by a polynucleotide comprising a nucleotide sequence, the nucleotide sequence hybridizing under stringent conditions to the complement of the sequence set forth in SEQ ID NO:1; and 
 (iii) a biologically active fragment of a GPCR of (i) or (ii); 
   (b) detecting a complex between the GPCR and the ligand;   (c) determining whether less of the complex is formed in the presence of the test compound than in the absence of the test compound;   (d) contacting the test compound in the presence of which less of the complex is formed in step (c) with a mammalian enteroendocrine cell in vitro; and   (e) determining whether the test compound stimulates GLP-1 secretion from the mammalian enteroendocrine cell,   wherein the ability of the test compound to stimulate GLP-1 secretion from the mammalian enteroendocrine cell identifies the test compound as a GLP-1 secretagogue.   
     
     
         63 . A method for identifying a GLP-1 secretagogue, comprising:
 (a) contacting a test compound with a host cell or with a membrane of a host cell that expresses a GPCR, wherein the GPCR comprises an amino acid sequence selected from the group consisting of:
 (i) amino acids 2-335 of SEQ ID NO:2; 
 (ii) the amino acid sequence of a GPCR encoded by a polynucleotide comprising a nucleotide sequence, the nucleotide sequence hybridizing under stringent conditions to the complement of the sequence set forth in SEQ ID NO:1; and 
 (iii)) a biologically active fragment of a GPCR of (i) or (ii); 
   (b) determining the ability of the test compound to stimulate functionality of the GPCR;   (c) administering the test compound that stimulates functionality of the GPCR in step (b) to a mammal; and   (d) determining whether the test compound increases a blood GLP-1 level in the mammal;   wherein the ability of the test compound to increase a blood GLP-1 level in the mammal identifies the test compound as a GLP-1 secretagogue.   
     
     
         64 . A method for identifying a GLP-1 secretagogue, comprising:
 (a) contacting a GPCR with a ligand to the GPCR in the presence of a test compound, wherein the GPCR comprises an amino acid sequence selected from the group consisting of:
 (i) amino acids 2-335 of SEQ ID NO:2; 
 (ii) the amino acid sequence of a GPCR encoded by a polynucleotide comprising a nucleotide sequence, the nucleotide sequence hybridizing under stringent conditions to the complement of the sequence set forth in SEQ ID NO:1; and 
 (iii) a biologically active fragment of a GPCR of (i) or (ii); 
   (b) detecting a complex between the GPCR and the ligand;   (c) determining whether less of the complex is formed in the presence of the test compound than in the absence of the test compound;   (d) determining the blood GLP-1 level in a biological sample obtained from a mammal, the mammal having been administered with a compound in the presence of which less of the complex is formed in step (c),   wherein the ability of the test compound to increase a blood GLP-1 level in the biological sample obtained from the mammal identifies the test compound as a GLP-1 secretagogue.   
     
     
         65 . A method of preparing a pharmaceutical composition comprising a GPR119 agonist having the effect of a GLP-1 secretagogue useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, the method comprising:
 (a) contacting the GPR119 agonist in vitro with a mammalian enteroendocrine cell; and   (b) determining whether the GPR119 agonist stimulates GLP-1 secretion from the mammalian enteroendocrine cell, wherein the ability of the GPR119 agonist to stimulate GLP-1 secretion from the mammalian enteroendocrine cell is indicative of the agonist being the GLP-1 secretagogue useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level; and   (c) admixing the GPR119 agonist with a pharmaceutically acceptable carrier.   
     
     
         66 . A method of preparing a pharmaceutical composition comprising a GPR119 agonist having the effect of a GLP-1 secretagogue, the GPR119 agonist having been contacted in vitro with a mammalian enteroendocrine cell and determined to stimulate GLP-1 secretion from the mammalian enteroendocrine cell, wherein the ability of the GPR119 agonist to stimulate GLP-1 secretion from the mammalian enteroendocrine cell is indicative of the agonist being the GLP-1 secretagogue, the method comprising admixing the GPR119 agonist with a pharmaceutically acceptable carrier. 
     
     
         67 . A method of preparing a pharmaceutical composition comprising a GPR119 agonist having the effect of a GLP-1 secretagogue useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, the method comprising:
 (a) determining a blood GLP-1 level in a biological sample obtained from a mammal, the mammal having been administered with the GPR119 agonist, wherein the ability of the GPR119 agonist to increase a blood GLP-1 level in the mammal is indicative of the agonist being the GLP-1 secretagogue useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level; and   (b) admixing the GPR119 agonist with a pharmaceutically acceptable carrier.   
     
     
         68 . A method of preparing a pharmaceutical composition comprising a GPR119 agonist having the effect of a GLP-1 secretagogue useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, the GPR119 agonist having been administered to a mammal and determined to increase a blood GLP-1 level in the mammal, wherein the ability of GPR119 agonist to increase a blood GLP-1 level is indicative of the agonist being the GLP-1 secretagogue useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, the method comprising admixing the GPR119 agonist with a pharmaceutically acceptable carrier. 
     
     
         69 . A method of preparing a dosage form of a pharmaceutical composition for increasing blood GLP-1 level in a subject, the pharmaceutical composition comprising a GPR119 agonist and a DPP-IV inhibitor, the method comprising:
 (a) admixing the GPR119 agonist with a pharmaceutically acceptable carrier to prepare a dosage form of the GPR119 agonist;   (b) admixing the DPP-IV inhibitor with a pharmaceutically acceptable carrier to prepare a dosage form of the DPP-IV inhibitor; and   (c) providing the dosage form of the GPR119 agonist and the dosage form of the DPP-IV inhibitor as a combined preparation for simultaneous, separate or sequential use.   
     
     
         70 . A combined preparation comprising a GPR119 agonist and a DPP-IV inhibitor, wherein the GPR119 agonist and the DPP-IV inhibitor are capable of being administered simultaneously, separately or sequentially, and wherein the DPP-IV inhibitor is not identical to 1-[2-[5-cyanopyridin-2-yl)amino]ethylamino]acetyl-2-cyano-(S)-pyrrolidine (NVP-DPP728). 
     
     
         71 . A twin pack comprising a GPR119 agonist and a DPP-IV inhibitor as a combined preparation for the treatment or prevention of diabetes or a condition related thereto. 
     
     
         72 . A method of treating diabetes or a condition related thereto in a subject in need thereof, comprising administering to the subject a therapeutically effective amount a GPR119 agonist and a DPP-IV inhibitor, wherein the DPP-IV inhibitor is not identical to 1-[2-[5-cyanopyridin-2-yl)amino]ethylamino]acetyl-2-cyano-(S)-pyrrolidine (NVP-DPP728). 
     
     
         73 . A method of treating a condition ameliorated by increasing a blood GLP-1 level in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a GPR119 agonist and a DPP-IV inhibitor, wherein the DPP-IV inhibitor is not identical to 1-[2-[5-cyanopyridin-2-yl)amino]ethylamino]acetyl-2-cyano-(S)-pyrrolidine (NVP-DPP728). 
     
     
         74 . A method of increasing a blood GLP-1 level in a subject deficient in GLP-1, comprising administering to the subject a therapeutically effective amount of a GPR119 agonist and a DPP-IV inhibitor, wherein the DPP-IV inhibitor is not identical to 1-[2-[5-cyanopyridin-2-yl)amino]ethylamino]acetyl-2-cyano-(S)-pyrrolidine (NVP-DPP728).