                         SEQUENCE LISTING

<110>  Sunshine Lake Pharma Co., Ltd.
 
<120>  GLP-1/GCG DUAL RECEPTOR AGONIST POLYPEPTIDE AND FUSION PROTEIN THEREOF

<130>  2021-12-2

<160>  18    

<170>  PatentIn version 3.5

<210>  1
<211>  29
<212>  PRT
<213>  Artificial

<220>
<223> Amino acid sequence of GLP-1/GCG dual receptor agonist polypeptide

<400>  1

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 
1               5                   10                  15      


Lys Ala Ala Lys Glu Phe Ile Glu Trp Leu Leu Glu Gly 
            20                  25                  


<210>  2
<211>  29
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of GLP-1/GCG dual receptor agonist polypeptide

<400>  2

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 
1               5                   10                  15      


Lys Arg Ala Lys Glu Phe Ile Glu Trp Leu Ile Ala Gly 
            20                  25                  


<210>  3
<211>  29
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of GLP-1/GCG dual receptor agonist polypeptide

<400>  3

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 
1               5                   10                  15      


Lys Arg Ala Lys Glu Phe Val Glu Trp Leu Ile Ala Gly 
            20                  25                  


<210>  4
<211>  29
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of GLP-1/GCG dual receptor agonist polypeptide

<400>  4

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 
1               5                   10                  15      


Lys Lys Ala Lys Glu Phe Ile Glu Trp Leu Ile Ala Gly 
            20                  25                  


<210>  5
<211>  29
<212>  PRT
<213>  Artificial

<220>
<223>   Amino acid sequence of GLP-1/GCG dual receptor agonist polypeptide

<400>  5

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 
1               5                   10                  15      


Lys Lys Ala Lys Glu Phe Val Glu Trp Leu Ile Ala Gly 
            20                  25                  


<210>  6
<211>  15
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of connecting peptide

<400>  6

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 
1               5                   10                  15  


<210>  7
<211>  16
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of connecting peptide

<400>  7

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala 
1               5                   10                  15      


<210>  8
<211>  10
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of connecting peptide

<400>  8

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 
1               5                   10  


<210>  9
<211>  228
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of IgG4-Fc fragment

<400>  9

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala 
1               5                   10                  15      


Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 
            20                  25                  30          


Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 
        35                  40                  45              


Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val 
    50                  55                  60                  


Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser 
65                  70                  75                  80  


Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 
                85                  90                  95      


Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser 
            100                 105                 110         


Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 
        115                 120                 125             


Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln 
    130                 135                 140                 


Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 
145                 150                 155                 160 


Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 
                165                 170                 175     


Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu 
            180                 185                 190         


Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser 
        195                 200                 205             


Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 
    210                 215                 220                 


Leu Ser Leu Gly 
225             


<210>  10
<211>  273
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of fusion protein

<400>  10

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 
1               5                   10                  15      


Lys Ala Ala Lys Glu Phe Ile Glu Trp Leu Leu Glu Gly Gly Gly Gly 
            20                  25                  30          


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Glu Ser Lys 
        35                  40                  45              


Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 
    50                  55                  60                  


Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 
65                  70                  75                  80  


Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 
                85                  90                  95      


Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 
            100                 105                 110         


Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 
        115                 120                 125             


Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 
    130                 135                 140                 


Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 
145                 150                 155                 160 


Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 
                165                 170                 175     


Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 
            180                 185                 190         


Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 
        195                 200                 205             


Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 
    210                 215                 220                 


Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp 
225                 230                 235                 240 


Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 
                245                 250                 255     


Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 
            260                 265                 270         


Gly 
    


<210>  11
<211>  273
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of fusion protein

<400>  11

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 
1               5                   10                  15      


Lys Arg Ala Lys Glu Phe Ile Glu Trp Leu Ile Ala Gly Gly Gly Gly 
            20                  25                  30          


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Glu Ser Lys 
        35                  40                  45              


Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 
    50                  55                  60                  


Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 
65                  70                  75                  80  


Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 
                85                  90                  95      


Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 
            100                 105                 110         


Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 
        115                 120                 125             


Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 
    130                 135                 140                 


Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 
145                 150                 155                 160 


Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 
                165                 170                 175     


Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 
            180                 185                 190         


Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 
        195                 200                 205             


Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 
    210                 215                 220                 


Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp 
225                 230                 235                 240 


Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 
                245                 250                 255     


Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 
            260                 265                 270         


Gly 
    


<210>  12
<211>  273
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of fusion protein

<400>  12

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 
1               5                   10                  15      


Lys Arg Ala Lys Glu Phe Val Glu Trp Leu Ile Ala Gly Gly Gly Gly 
            20                  25                  30          


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Glu Ser Lys 
        35                  40                  45              


Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 
    50                  55                  60                  


Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 
65                  70                  75                  80  


Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 
                85                  90                  95      


Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 
            100                 105                 110         


Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 
        115                 120                 125             


Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 
    130                 135                 140                 


Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 
145                 150                 155                 160 


Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 
                165                 170                 175     


Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 
            180                 185                 190         


Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 
        195                 200                 205             


Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 
    210                 215                 220                 


Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp 
225                 230                 235                 240 


Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 
                245                 250                 255     


Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 
            260                 265                 270         


Gly 
    


<210>  13
<211>  273
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of fusion protein

<400>  13

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 
1               5                   10                  15      


Lys Lys Ala Lys Glu Phe Ile Glu Trp Leu Ile Ala Gly Gly Gly Gly 
            20                  25                  30          


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Glu Ser Lys 
        35                  40                  45              


Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 
    50                  55                  60                  


Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 
65                  70                  75                  80  


Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 
                85                  90                  95      


Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 
            100                 105                 110         


Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 
        115                 120                 125             


Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 
    130                 135                 140                 


Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 
145                 150                 155                 160 


Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 
                165                 170                 175     


Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 
            180                 185                 190         


Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 
        195                 200                 205             


Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 
    210                 215                 220                 


Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp 
225                 230                 235                 240 


Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 
                245                 250                 255     


Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 
            260                 265                 270         


Gly 
    


<210>  14
<211>  273
<212>  PRT
<213>  Artificial

<220>
<223>  Amino acid sequence of fusion protein

<400>  14

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Glu 
1               5                   10                  15      


Lys Lys Ala Lys Glu Phe Val Glu Trp Leu Ile Ala Gly Gly Gly Gly 
            20                  25                  30          


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Glu Ser Lys 
        35                  40                  45              


Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 
    50                  55                  60                  


Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 
65                  70                  75                  80  


Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu 
                85                  90                  95      


Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 
            100                 105                 110         


Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg 
        115                 120                 125             


Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 
    130                 135                 140                 


Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu 
145                 150                 155                 160 


Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 
                165                 170                 175     


Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu 
            180                 185                 190         


Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 
        195                 200                 205             


Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 
    210                 215                 220                 


Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp 
225                 230                 235                 240 


Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His 
                245                 250                 255     


Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu 
            260                 265                 270         


Gly 


<210>  15
<211>  39
<212>  PRT
<213>  Artificial

<220>
<223>  Sequence of control group LY3298176Xaa can be Aib

<400>  15

Tyr Xaa Glu Gly Thr Phe Thr Ser Asp Tyr Ser Ile Xaa Leu Asp Lys 
1               5                   10                  15      


Ile Ala Gln Lys Ala Phe Val Gln Trp Leu Ile Ala Gly Gly Pro Ser 
            20                  25                  30          


Ser Gly Ala Pro Pro Pro Ser 
        35                  


<210>  16
<211>  30
<212>  PRT
<213>  Artificial

<220>
<223>  Sequence of control group MEDI0382 

<400>  16

His Ser Gln Gly Thr Phe Thr Ser Asp Lys Ser Glu Tyr Leu Asp Ser 
1               5                   10                  15      


Glu Arg Ala Arg Asp Phe Val Ala Trp Leu Glu Ala Gly Gly 
            20                  25                  30  


<210>  17
<211>  31
<212>  PRT
<213>  

<220>
<223>  Human GLP-1
 
<400>  17

His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly 
1               5                   10                  15      


Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly 
            20                  25                  30      


<210>  18
<211>  29
<212>  PRT
<213>  

<220>
<223>  Human GCG

<400>  18

His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser 
1               5                   10                  15      


Arg Arg Ala Gln Asp Phe Val Gln Trp Leu Met Asn Thr 
            20                  25       
