                         SEQUENCE LISTING

<110>  Novo Nordisk A/S
 
<120>  STABLE GLUCAGON ANALOGUES AND USE FOR TREATMENT OF HYPOGLYCAEMIA

<130>  130077WO01

<160>  24    

<170>  PatentIn version 3.5

<210>  1
<211>  29
<212>  PRT
<213>  HOMO SAPIENS

<400>  1

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                  


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

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  This amino acid residue is alpha-aminoisobutyric acid (Aib)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  2

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


Arg Arg Ala Gln Asp Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


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

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  This amino acid residue is alpha-aminoisobutyric acid (Aib)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  3

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


Arg Arg Ala Gln Asp Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


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

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  This amino acid residue is alpha-aminoisobutyric acid (Aib)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  4

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


Arg Arg Ala Gln Asp Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


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

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  This amino acid residue is alpha-aminoisobutyric acid (Aib)

<220>
<221>  MISC_FEATURE
<222>  (28)..(28)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  5

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


Arg Arg Ala Gln Asp Phe Val Ala Trp Leu Leu Lys Thr 
            20                  25                  


<210>  6
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  6

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


Arg Arg Ala Gln Asp Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  7
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  7

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


Arg Arg Ala Gln Glu Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  8
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  This amino acid residue is alpha-aminoisobutyric acid (Aib)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  8

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


Arg Arg Ala Gln Glu Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  9
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  9

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


Arg Arg Ala Gln Glu Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  10
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  10

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


Arg Arg Ala Gln Asp Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  11
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  This amino acid residue is alpha-aminoisobutyric acid (Aib)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  11

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


Arg Arg Ala Gln Asp Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  12
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (16)..(16)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  12

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


Arg Arg Ala Gln Glu Phe Val Gln Trp Leu Leu Ser Thr 
            20                  25                  


<210>  13
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (20)..(20)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  13

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


Arg Arg Ala Lys Glu Phe Val Gln Trp Leu Leu Ser Thr 
            20                  25                  


<210>  14
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (21)..(21)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  14

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


Arg Arg Ala Gln Lys Phe Val Gln Trp Leu Leu Ser Thr 
            20                  25                  


<210>  15
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (28)..(28)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  15

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


Arg Arg Ala Gln Glu Phe Val Gln Trp Leu Leu Lys Thr 
            20                  25                  


<210>  16
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (29)..(29)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  16

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


Arg Arg Ala Gln Glu Phe Val Gln Trp Leu Leu Ser Lys 
            20                  25                  


<210>  17
<211>  30
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (30)..(30)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  17

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


Arg Arg Ala Gln Glu Phe Val Gln Trp Leu Leu Ser Thr Lys 
            20                  25                  30  


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

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[2-[[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carb
       oxybutanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]ami
       no]-4-carboxybutanoyl]amino]acetyl]-Lys

<400>  18

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


Arg Arg Ala Gln Glu Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  19
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxybutanoyl]a
       mino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]-Lys

<400>  19

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


Arg Arg Ala Gln Glu Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  20
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4
       -carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoy
       l]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]-Lys

<400>  20

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


Arg Arg Ala Gln Glu Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  21
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  21

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


Arg Arg Ala Gln Asp Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  22
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  misc_feature
<222>  (16)..(16)
<223>  Xaa can be any naturally occurring amino acid

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  22

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


Arg Arg Ala Gln Asp Phe Val Lys Trp Leu Leu Ser Thr 
            20                  25                  


<210>  23
<211>  29
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial protein based on human glucagon


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  This amino acid residue is modified to imidazopropionyl (Imp)

<220>
<221>  MISC_FEATURE
<222>  (24)..(24)
<223>  This amino acid residue is 
       N-epsilon-[(4S)-4-[[(4S)-4-[[(4S)-4-[[(4S)-4-acetamido-4-carboxyb
       utanoyl]amino]-4-carboxybutanoyl]amino]-4-carboxybutanoyl]amino]-
       4-carboxybutanoyl]-Lys

<400>  23

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


Arg Arg Ala Gln Glu Phe Val Lys Trp Leu Leu Glu Thr 
            20                  25                  


<210>  24
<211>  30
<212>  PRT
<213>  ARTIFICIAL

<220>
<223>  Artificial sequence based on human glucagon


<220>
<221>  SITE
<222>  (1)..(1)
<223>  This residue is Imp (desaminohistidine)

<220>
<221>  SITE
<222>  (2)..(2)
<223>  This residue is Ser or Aib

<220>
<221>  SITE
<222>  (10)..(10)
<223>  This residue is Tyr, Leu, Ile or Val

<220>
<221>  SITE
<222>  (12)..(12)
<223>  This residue is Lys or Arg

<220>
<221>  SITE
<222>  (15)..(15)
<223>  This residue is Asp or Glu

<220>
<221>  SITE
<222>  (16)..(16)
<223>  This residue is Ser, Ala, Leu, Thr, Aib, Ile, Val or Lys

<220>
<221>  SITE
<222>  (20)..(20)
<223>  This residue is Gln, Glu, Aib or Lys

<220>
<221>  SITE
<222>  (21)..(21)
<223>  This residue is Asp, Glu or Lys

<220>
<221>  SITE
<222>  (24)..(24)
<223>  This residue is Gln, Ala, Glu, Aib or Lys

<220>
<221>  SITE
<222>  (27)..(27)
<223>  This residue is Met, Leu or Val

<220>
<221>  SITE
<222>  (28)..(28)
<223>  This residue is Asn, Ser, Thr, Gln, Ala, Gly, Glu or Lys

<220>
<221>  SITE
<222>  (29)..(29)
<223>  This residue is Thr, Gly, Ser, Gln, Ala, Glu or Lys

<220>
<221>  SITE
<222>  (30)..(30)
<223>  This residue is absent or is Lys

<400>  24

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


Arg Arg Ala Xaa Xaa Phe Val Xaa Trp Leu Xaa Xaa Xaa Xaa 
            20                  25                  30  


