Novel Acylated Insulin Analogues and Uses Thereof

Abstract

The present invention relates to novel insulin analogues and derivatives thereof, such as acylated insulin analogues, and their pharmaceutical use, in particular in the treatment or prevention of medical conditions relating to diabetes, obesity and cardiovascular diseases.

1 . An insulin derivative, comprising an insulin analogue comprising B5Y or B5F and a substituent comprising an acyl group, or a pharmaceutically acceptable salt, amide or ester thereof. 
     
     
         2 . The insulin derivative according to  claim 1 , wherein said insulin analogue further comprises B26G or B26A. 
     
     
         3 . The insulin derivative according to  claim 1 , wherein said insulin analogue further comprises B28K, B26K or B29 K and said acyl group is attached to B28K, B26K or B29K. 
     
     
         4 . The insulin derivative according to  claim 2 , wherein said insulin analogue further comprises B28K, B26K or B29 K and said acyl group is attached to B28K, B26K or B29K. 
     
     
         5 . The insulin derivative according to  claim 1 , wherein said substituent has the following formula (I):
   Acy-L1-L2-L3   
       wherein:
 Acy is an acyl group and is represented by lithocholic acid or comprises at least one functional group of formulae:
   —CO—(CH 2 ) x —COOH; or  Chem. 1:
 
   —CO—(CH 2 ) x -tetrazolyl;  Chem. 2:
 
 
 
       wherein x represents an integer in the range of from 12 to 20; and the tetrazolyl group is 1H-tetrazol-5-yl. 
       or is a fatty acid of formula:
   —CO—(CH 2 ) x —CH 3   Chem. 3:
 
 
       wherein x represents an integer in the range from 8 to 16,
 L1 is absent and represents a covalent bond or represents OEG, gGlu, DgGlu or sulfonimide C-4 
 L2 is absent and represents a covalent bond or represents OEG, gGlu, DgGlu or sulfonimide C-4 
 L3 is absent and represents a covalent bond or represents OEG, gGlu, DgGlu or sulfonimide C-4 
 
       wherein gGlu represents a gamma glutamic acid residue and OEG represents [2-(2-aminoethoxy)ethoxy]acetyl. 
     
     
         6 . The insulin derivative according to  claim 1 , wherein said insulin analogue further comprises A14E and/or desB30 and/or desB29-30 and/or desB27-30. 
     
     
         7 . The insulin derivative according to  claim 1 , wherein said insulin analogue comprises
 i. A14E, B5Y, B26A, B28K, desB29-30;   ii. A14E, B5Y, B26G, B28K, desB29-30;   iii. B5Y, B26A, B28K, desB29-30;   iv. B5Y, B26G, B28K, desB29-30;   v. B5Y, B28K, desB29-30;   vi. A14E, B5F, B26G, B28K, desB29-30;   vii. B5F, B28K, desB29-30;   viii. B5Y, B26K, desB27-desB30;   ix. B5Y, desB30;   x. B5Y, B26G, desB30;   xi. B5Y, B26A, desB30;   xii. B5F, B26A, B28K, desB29-30;   xiii. B5Y, B26A, B28K, desB29-30;   xiv. B5Y, B26G, B28K, desB29-30;   xv. B5F, B26G, B28K, desB29-30;   xvi. A14E, B5F, B26A, B28K, desB29-30;   xvii. A14E, B5Y, B26A, B28K, desB29-30;   xviii. A14E, B5Y, B26G, B28K, desB29-30;   xix. A14E, B5Y, B28K, desB29-30;   xx. A14E, B5F, B28K, desB29-30;   xxi. A14E, B5Y, B26K, desB27-desB30;   xxii. A14E, B5Y, desB30;   xxiii. A14E, B5Y, B26G, desB30; or   xxiv. A14E, B5Y, B26A, desB30.   
     
     
         8 . The insulin derivative according to  claim 5 , wherein said insulin analogue comprises
 i. A14E, B5Y, B26A, B28K, desB29-30;   ii. A14E, B5Y, B26G, B28K, desB29-30;   iii. B5Y, B26A, B28K, desB29-30;   iv. B5Y, B26G, B28K, desB29-30;   v. B5Y, B28K, desB29-30;   vi. A14E, B5F, B26G, B28K, desB29-30;   vii. B5F, B28K, desB29-30;   viii. B5Y, B26K, desB27-desB30;   ix. B5Y, desB30;   x. B5Y, B26G, desB30;   xi. B5Y, B26A, desB30;   xii. B5F, B26A, B28K, desB29-30;   xiii. B5Y, B26A, B28K, desB29-30;   xiv. B5Y, B26G, B28K, desB29-30;   xv. B5F, B26G, B28K, desB29-30;   xvi. A14E, B5F, B26A, B28K, desB29-30;   xvii. A14E, B5Y, B26A, B28K, desB29-30;   xviii. A14E, B5Y, B26G, B28K, desB29-30;   xix. A14E, B5Y, B28K, desB29-30;   xx. A14E, B5F, B28K, desB29-30;   xxi. A14E, B5Y, B26K, desB27-desB30;   xxii. A14E, B5Y, desB30;   xxiii. A14E, B5Y, B26G, desB30; or   xxiv. A14E, B5Y, B26A, desB30.   
     
     
         9 . The insulin derivative according to  claim 7 , wherein Acy is selected from the group consisting of: lithocholic acid, 1,16-hexadecanedioic acid, 1,18-octadecanedioic acid, 1,20-eicosanedioic acid, tetrazole-C16, tetrazole-C17, tetrazole C18 and tetradecanoic acid. 
     
     
         10 . The insulin derivative according to  claim 8 , wherein -L1-L2-L3 represents a divalent linking group selected from group consisting of DgGlu, gGlu, gGlu-gGlu, gGlu-OEG, gGlu-OEG-OEG, OEG, sulfonimide-C4, and sulfonimide-C4-sulfonimide-C4. 
     
     
         11 . The insulin derivative according to  claim 1 , wherein said insulin derivative is N{Epsilon-B28}-[2-[2-[2-[[(4S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butanoyl]amino]ethoxy]ethoxy]acetyl]-[GluA14,TyrB5,GlyB26,LysB28],des-(B29-B30)-lnsulin 
       
         
           
           
               
               
           
         
       
     
     
         12 . An insulin analogue comprising
 i. A14E, B5Y, B26A, B28K, desB29-30;   ii. A14E, B5Y, B26G, B28K, desB29-30;   iii. B5Y, B26A, B28K, desB29-30;   iv. B5Y, B26G, B28K, desB29-30;   v. B5Y, B28K, desB29-30;   vi. A14E, B5F, B26G, B28K, desB29-30;   vii. B5F, B28K, desB29-30;   viii. B5Y, B26K, desB27-desB30;   ix. B5Y, desB30;   x. B5Y, B26G, desB30;   xi. B5Y, B26A, desB30;   xii. B5F, B26A, B28K, desB29-30;   xiii. B5Y, B26A, B28K, desB29-30;   xiv. B5Y, B26G, B28K, desB29-30;   xv. B5F, B26G, B28K, desB29-30;   xvi. A14E, B5F, B26A, B28K, desB29-30;   xvii. A14E, B5Y, B26A, B28K, desB29-30;   xviii. A14E, B5Y, B26G, B28K, desB29-30;   xix. A14E, B5Y, B28K, desB29-30;   xx. A14E, B5F, B28K, desB29-30;   xxi. A14E, B5Y, B26K, desB27-desB30;   xxii. A14E, B5Y, desB30;   xxiii. A14E, B5Y, B26G, desB30; or   xxiv. A14E, B5Y, B26A, desB30.   
       or a pharmaceutically acceptable salt, amide or ester thereof. 
     
     
         13 . An insulin derivative according to  claim 1 , wherein said insulin analogue comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, and SEQ ID NOs: 3-15. 
     
     
         14 . An insulin derivative according to  claim 5 , wherein said insulin analogue comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, and SEQ ID NOs: 3-15. 
     
     
         15 . A method of treating diabetes, cardiovascular disease, atherosclerosis, or endothelial dysfunction, or reducing liver triglyceride content or reducing body weight gain, comprising administering to a subject in need thereof a therapeutically effective amount of an insulin derivative according to  claim 1 . 
     
     
         16 . A method of treating diabetes, cardiovascular disease, atherosclerosis, or endothelial dysfunction, or reducing liver triglyceride content or reducing body weight gain, comprising administering to a subject in need thereof a therapeutically effective amount of an insulin derivative according to  claim 8 . 
     
     
         17 . A method for treating diabetes, diabetes of Type 1, diabetes of Type 2, impaired glucose tolerance, hyperglycemia, dyslipidemia, obesity, metabolic syndrome X, insulin resistance syndrome, hypertension, cognitive disorders, atherosclerosis, myocardial infarction, stroke, cardiovascular disorders, coronary heart disease, stroke, inflammatory bowel syndrome, dyspepsia, hypotension or gastric ulcers, comprising administrating to a subject in need thereof a therapeutically effective amount of an insulin derivative according to  claim 1 . 
     
     
         18 . A method for treating diabetes, diabetes of Type 1, diabetes of Type 2, impaired glucose tolerance, hyperglycemia, dyslipidemia, obesity, metabolic syndrome X, insulin resistance syndrome, hypertension, cognitive disorders, atherosclerosis, myocardial infarction, stroke, cardiovascular disorders, coronary heart disease, stroke, inflammatory bowel syndrome, dyspepsia, hypotension or gastric ulcers, comprising administrating to a subject in need thereof a therapeutically effective amount of an insulin derivative according to  claim 8 . 
     
     
         19 . A method for treating diabetes, diabetes of Type 1, diabetes of Type 2, impaired glucose tolerance, hyperglycemia, dyslipidemia, obesity, metabolic syndrome X, insulin resistance syndrome, hypertension, cognitive disorders, atherosclerosis, myocardial infarction, stroke, cardiovascular disorders, coronary heart disease, stroke, inflammatory bowel syndrome, dyspepsia, hypotension or gastric ulcers, comprising administrating to a subject in need thereof a therapeutically effective amount of an insulin analogue according to  claim 11 . 
     
     
         20 . A method for determining selectivity of an insulin compound comprising the following steps:
 measuring the maximal AKT phosphorylation induced by said insulin compound relative to human insulin   measuring the maximal ERK activation induced by said insulin compound relative to human insulin,   
       wherein the ERK/AKT ratio is less than 1.