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 . A method of treating or preventing diabetes or a condition related thereto comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising a GPR119 agonist and a DPP-IV inhibitor.  
     
     
         2 . A method in accordance with  claim 1 , wherein the GPR119 agonist and the DPP-IV inhibitor are administered in amounts sufficient to lower a blood glucose level in the subject.  
     
     
         3 . A method of treating or preventing a condition ameliorated by increasing a blood GLP-1 level comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising a GPR119 agonist and a DPP-IV inhibitor.  
     
     
         4 . A method in accordance with  claim 3 , wherein the GPR119 agonist and the DPP-IV inhibitor are administered in amounts sufficient to increase a blood GLP-1 level in the subject.  
     
     
         5 . A method of increasing a blood GLP-1 level comprising administering to a subject deficient in GLP-1 a therapeutically effective amount of a composition comprising a GPR119 agonist and a DPP-IV inhibitor.  
     
     
         6 . A method in accordance with  claim 5 , wherein the GPR119 agonist and the DPP-IV inhibitor are administered in amounts sufficient to increase a blood GLP-1 level in the subject.  
     
     
         7 . A method in accordance with  claim 1  or  claim 2 , wherein diabetes is Type 2 diabetes.  
     
     
         8 . A method in accordance with  claim 1  or  claim 2 , wherein the condition related to diabetes is selected from the group consisting of hyperglycemia, impaired glucose tolerance, insulin resistance, pancreatic beta-cell insufficiency, enteroendocrine cell insufficiency, glucosuria, metabolic acidosis, cataracts, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, diabetic coronary artery disease, diabetic cerebrovascular disease, diabetic peripheral vascular disease, metabolic syndrome, hyperlipidemia, atherosclerosis, stroke, hypertension, and obesity.  
     
     
         9 . A method in accordance with  claim 3  or  claim 4 , wherein the condition ameliorated by increasing a blood GLP-1 level is selected from the group consisting of diabetes, a condition related to diabetes, myocardial infarction, learning impairment, memory impairment, and a neurodegenerative disorder.  
     
     
         10 . A method in accordance with  claim 3  or  claim 4 , wherein the condition ameliorated by increasing a blood GLP-1 level is a neurodegenerative disorder selected from the group consisting of excitotoxic brain damage caused by severe epileptic seizures, Alzheimer's disease, Parkinson's disease, Huntington's disease, prion-associated disease, stroke, motor-neuron disease, learning or memory impairment, traumatic brain injury, spinal cord injury, and peripheral neuropathy.  
     
     
         11 . A method in accordance with any one of claims  1 ,  3 , and  5 , wherein the subject is a human.  
     
     
         12 . A composition comprising a GPR119 agonist and a DPP-IV inhibitor.  
     
     
         13 . A dosage form of a composition in accordance with  claim 12 , wherein the GPR119 agonist and the DPP-IV inhibitor are in amounts sufficient to lower a blood glucose level in a subject.  
     
     
         14 . A dosage form of a composition in accordance with  claim 12 , wherein the GPR119 agonist and the DPP-IV inhibitor are in amounts sufficient to increase a blood GLP-1 level in a subject.  
     
     
         15 . A dosage form in accordance with  claim 13  or  14 , where the subject is human.  
     
     
         16 . A composition comprising a GPR119 agonist and a DPP-IV inhibitor, for use in a method of treatment of the human or animal body by therapy.  
     
     
         17 . A composition in accordance with  claim 16 , wherein the GPR119 agonist and the DPP-IV inhibitor are in amounts sufficient to lower a blood glucose level in a subject.  
     
     
         18 . A composition in accordance with  claim 16 , wherein the GPR119 agonist and the DPP-IV inhibitor are in amounts to increase a blood GLP-1 level in a subject.  
     
     
         19 . A method of preparing a pharmaceutical composition, said method comprising admixing a GPR119 agonist and a DPP-IV inhibitor, together with at least one pharmaceutically acceptable carrier.  
     
     
         20 . A method in accordance with  claim 19 , said method further comprising the step of preparing a dosage form of the pharmaceutical composition wherein the GPR119 agonist and the DPP-IV inhibitor are in amounts sufficient to lower a blood glucose level in a subject.  
     
     
         21 . A method in accordance with  claim 19 , said method further comprising the step of preparing a dosage form of the pharmaceutical composition wherein the GPR119 agonist and the DPP-IV inhibitor are in amounts sufficient to increase a blood GLP-1 level in a subject.  
     
     
         22 . A pharmaceutical composition comprising a GPR119 agonist and a DPP-IV inhibitor, together with at least one pharmaceutically acceptable carrier.  
     
     
         23 . A dosage form of a pharmaceutical composition in accordance with  claim 22 , wherein the GPR119 agonist and the DPP-IV inhibitor are in amounts sufficient to lower a blood glucose level in a subject.  
     
     
         24 . A dosage form of a pharmaceutical composition in accordance with  claim 22 , wherein the GPR119 agonist and the DPP-IV inhibitor are in amounts sufficient to increase a blood GLP-1 level in a subject.  
     
     
         25 . A method of treating or preventing diabetes or a condition related thereto comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition in accordance with  claim 22  or  claim 23 .  
     
     
         26 . A method in accordance with  claim 25 , wherein the GPR119 agonist and the DPP-IV inhibitor are in amounts sufficient to lower a blood glucose level in the subject.  
     
     
         27 . A method of treating or preventing a condition ameliorated by increasing a blood GLP-1 level comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition in accordance with  claim 22  or  claim 24 .  
     
     
         28 . A method of increasing a blood GLP-1 level comprising administering to a subject deficient in GLP-1 a therapeutically effective amount of a pharmaceutical composition in accordance with  claim 22  or  claim 24 .  
     
     
         29 . A method in accordance with  claim 27  or  claim 28 , wherein the GPR119 agonist and the DPP-IV inhibitor are in amounts sufficient to increase a blood GLP-1 level in the subject.  
     
     
         30 . A method in accordance with  claim 20  or  21 , wherein the subject is a human.  
     
     
         31 . A method in accordance with  claim 25 , wherein the subject is human.  
     
     
         32 . A method in accordance with  claim 27 , wherein the subject is human.  
     
     
         33 . A method in accordance with  claim 28 , wherein the subject is human  
     
     
         34 . A dosage form in accordance with  claim 23  or  24 , wherein the subject is a human.  
     
     
         35 . A method for identifying GLP-1 secretagogues or compounds useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, comprising the steps of: 
 (a) contacting a test compound with a host cell or with membrane of a host cell that expresses a G protein-coupled receptor, wherein the G protein-coupled receptor comprises an amino acid sequence selected from the group consisting of: 
 (i) amino acids 1-335 of SEQ ID NO:2;  
 (ii) amino acids 2-335 of SEQ ID NO:2;  
 (iii) amino acids 2-335 of SEQ ID NO:2, with the proviso that the receptor does not comprise the amino acid sequence of SEQ ID NO:2;  
 (iv) the amino acid sequence of a G protein-coupled receptor encoded by a polynucleotide comprising a nucleotide sequence, said nucleotide sequence being the sequence obtainable by a process comprising performing polymerase chain reaction (PCR) on a human DNA sample using specific primers SEQ ID NO:3 and SEQ ID NO:4;  
 (v) the amino acid sequence of a G protein-coupled receptor encoded by a polynucleotide comprising a nucleotide sequence, said nucleotide sequence hybridizing under stringent conditions to the complement of SEQ ID NO:1; and  
 (vi) a biologically active fragment of any one of (i) to (v); and  
   (b) determining the ability of the test compound to stimulate functionality of the receptor;    wherein the ability of the test compound to stimulate functionality of the receptor is indicative of the test compound being a GLP-1 secretagogue or a compound useful for preventing or treating a condition ameliorated by increasing a blood GLP-1 level.    
     
     
         36 . A method for identifying GLP-1 secretagogues or compounds useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, comprising the steps of the method of  claim 35 , and further comprising: 
 (c) contacting a compound which stimulates functionality of the receptor in step (b) in vitro with a mammalian enteroendocrine cell; and    (d) determining whether the 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 is indicative of the test compound being a GLP-1 secretagogue or a compound useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level.    
     
     
         37 . A method for identifying GLP-1 secretagogues or compounds useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, comprising the steps of the method of  claim 35 , and further comprising: 
 (c) administering a compound which stimulates functionality of the receptor in step (b) to a mammal; and    (d) determining whether the 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 is indicative of the test compound being a GLP-1 secretagogue or a compound useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level.    
     
     
         38 . A method in accordance with  claim 37 , wherein the mammal is a non-human mammal.  
     
     
         39 . A method in accordance with any one of  claims 35  to  38 , wherein the host cell comprises an expression vector, said expression vector comprising a polynucleotide encoding the G protein-coupled receptor.  
     
     
         40 . A method in accordance with  claim 35 , wherein said determining 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+.  
     
     
         41 . A method in accordance with  claim 40 , wherein a level of cAMP is increased.  
     
     
         42 . A method in accordance with  claim 35 , wherein said determining is through the use of a Melanophore assay, or through the measurement of GTPγS binding to a membrane comprising said GPCR.  
     
     
         43 . A method for identifying GLP-1 secretagogues or compounds useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, comprising the steps of: 
 (a) contacting a G protein-coupled receptor with an optionally labeled known ligand to the receptor in the presence or absence of a test compound, wherein the G protein-coupled receptor comprises an amino acid sequence selected from the group consisting of: 
 (i) amino acids 1-335 of SEQ ID NO:2;  
 (ii) amino acids 2-335 of SEQ ID NO:2;  
 (iii) amino acids 2-335 of SEQ ID NO:2, with the proviso that the receptor does not comprise the amino acid sequence of SEQ ID NO:2;  
 (iv) the amino acid sequence of a G protein-coupled receptor encoded by a polynucleotide comprising a nucleotide sequence, said nucleotide sequence being the sequence obtainable by a process comprising performing polymerase chain reaction (PCR) on a human DNA sample using specific primers SEQ ID NO:3 and SEQ ID NO:4;  
 (v) the amino acid sequence of a G protein-coupled receptor encoded by a polynucleotide comprising a nucleotide sequence, said nucleotide sequence hybridizing under stringent conditions to the complement of SEQ ID NO:1; and  
 (vi) a biologically active fragment of any one of (i) to (v); and  
   (b) detecting the complex between said known ligand and said receptor; and    (c) determining whether less of said complex is formed in the presence of the test compound than in the absence of the test compound;    wherein said determination is indicative of the test compound being a GLP-1 secretagogue or a compound useful for preventing or treating a condition ameliorated by increasing a blood GLP-1 level.    
     
     
         44 . A method for identifying GLP-1 secretagogues or compounds useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, comprising the steps of the method of  claim 43 , and further comprising: 
 (d) contacting a compound in the presence of which less of said complex is formed in step (c) in vitro with a mammalian enteroendocrine cell; and    (e) determining whether the 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 is indicative of the test compound being a GLP-1 secretagogue or a compound useful for preventing or treating a condition ameliorated by increasing a blood GLP-1 level.    
     
     
         45 . A method for identifying GLP-1 secretagogues or compounds useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level, comprising the steps of the method of  claim 43 , and further comprising: 
 (d) administering a compound in the presence of which less of said complex is formed in step (c) to a mammal; and    (e) determining whether the 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 is indicative of the test compound being a GLP-1 secretagogue or a compound useful for treating or preventing a condition ameliorated by increasing a blood GLP-1 level.    
     
     
         46 . A method in accordance with  claim 45 , wherein the mammal is a non-human mammal.  
     
     
         47 . The method in accordance with any one of  claims 43  to  46 , wherein the receptor is recombinant.