U.S. patent application number 11/454804 was filed with the patent office on 2007-02-22 for novel glp-1 compounds.
This patent application is currently assigned to Novo Nordisk A/S. Invention is credited to Thomas Kruse Hansen, Nils Langeland Johansen, Jesper Lau, Kjeld Madsen, Jeppe Sturis.
Application Number | 20070042956 11/454804 |
Document ID | / |
Family ID | 34702152 |
Filed Date | 2007-02-22 |
United States Patent
Application |
20070042956 |
Kind Code |
A1 |
Johansen; Nils Langeland ;
et al. |
February 22, 2007 |
Novel GLP-1 compounds
Abstract
Novel GLP-1 compounds and their therapeutic use.
Inventors: |
Johansen; Nils Langeland;
(Kobenhavn O, DK) ; Lau; Jesper; (Farum, DK)
; Madsen; Kjeld; (Vaerlose, DK) ; Hansen; Thomas
Kruse; (Herlev, DK) ; Sturis; Jeppe;
(Vaerlose, DK) |
Correspondence
Address: |
NOVO NORDISK, INC.;PATENT DEPARTMENT
100 COLLEGE ROAD WEST
PRINCETON
NJ
08540
US
|
Assignee: |
Novo Nordisk A/S
Bagsvaerd
DK
|
Family ID: |
34702152 |
Appl. No.: |
11/454804 |
Filed: |
June 16, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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PCT/DK04/00886 |
Dec 17, 2004 |
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11454804 |
Jun 16, 2006 |
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Current U.S.
Class: |
514/4.9 ;
514/11.7; 514/18.9; 514/6.7; 514/6.8; 530/324 |
Current CPC
Class: |
A61P 9/00 20180101; A61K
38/00 20130101; C07K 14/57563 20130101; A61P 9/10 20180101; A61P
25/28 20180101; A61P 9/12 20180101; A61P 25/00 20180101; C07K
14/605 20130101; A61P 3/10 20180101; A61P 5/48 20180101; A61P 3/04
20180101; A61P 1/04 20180101; A61P 3/00 20180101; A61P 3/06
20180101; A61P 1/14 20180101; A61P 1/00 20180101 |
Class at
Publication: |
514/012 ;
530/324 |
International
Class: |
A61K 38/22 20070101
A61K038/22; C07K 14/575 20070101 C07K014/575 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 18, 2003 |
DK |
PA 2003 01885 |
Jul 9, 2004 |
DK |
PA 2004 01090 |
Claims
1. A compound having a structure of formula (I) Insulinotropic
agent(--Y--C*).sub.f-Q (I) wherein Insulinotropic agent is a
radical derived from an insulinotropic peptide which binds to the
human glucagon-like peptide 1 (GLP-1) receptor, or a radical
derived from a peptide in which 22 positions out of the first 30
are identical to those found in corresponding positions in GLP-1 or
found in corresponding positions in Exendin-4, and Y is a bivalent
connecting chemical group connecting C* with the Insulinotropic
agent, and C* is a bivalent polar separating chemical group where
50-20% of the heavy atoms are either O or N, and f is 0 or 1 and Q
is selected from ##STR25## wherein A is a polar chemical group of a
single molecular size (monodisperse) or of several molecular sizes
(polydisperse) where 50-20% of the heavy atoms are independently
oxygen or nitrogen, and W is a bivalent chemical group whereby A is
connected, and X is a bivalent connecting chemical group whereby B
is connected, and B is a connecting or branching chemical
group.
2. The compound according to claim 1, wherein the C-terminal amino
acid residue of said insulinotropic agent is not cysteine.
3. The compound according to claim 1, wherein A is a monodisperse
or polydisperse chemical group having the structure
--(CH.sub.2).sub.lO[(CH.sub.2).sub.nO].sub.m(CH.sub.2).sub.p--H,
where l, n and p independently are an integer in the range from 1
to 10, m is an integer in the range from 1 to 5000, and where m
multiplied by n+1 is less than 10000.
4. The compound according to claim 1, wherein A is a monodisperse
or polydisperse chemical group having the structure
--(CH.sub.2).sub.lC(.dbd.O)O[(CH.sub.2).sub.nO].sub.m(CH.sub.2).sub.p--H,
where l, n and p independently are an integer in the range from 1
to 10, m is an integer in the range from 1 to 5000, and where m
multiplied by n+1 is less than 10000.
5. The compound according to claim 3, wherein n is 2 or 3.
6. The compound according to claim 3, wherein m is in the range
from 10-1000, or in the range from 20-250.
7. The compound according to claim 1, wherein A is a monodisperse
or polydisperse chemical group having the structure
-(Z.sup.1(CH.sub.2).sub.lO[(CH.sub.2).sub.2O].sub.m(CH.sub.2).sub.p--NR.s-
up.1).sub.q-Z.sup.2, where Z.sup.1 is --CO-- or
--CO--(CH.sub.2).sub.n--CO--NH--, and Z.sup.2 is --R.sup.1,
--CO--(CH.sub.2).sub.n--R.sup.1,
--(CH.sub.2).sub.lO[(CH.sub.2).sub.2O].sub.m(CH.sub.2).sub.P--R.sup.1
wherein l and n and p independently are integers in the range from
1 to 10, and R.sup.1 is --OH, --NH.sub.2, --NH--R.sup.2,
--NH(--R.sup.2)--R.sup.2, --COOH, C.sub.1-6-alkyl, or
--NH--CH(R.sup.2)--COOH, and where m and q are independently
integers in the range from 1 to 20, and where l, n and p are
independently integers in the range from 1 to 6, and R.sup.2 is
hydrogen or C.sub.1-6alkyl.
8. The compound according to claim 1, wherein A is mPEGyl.
9. The compound according to claim 1, wherein A is
mPEGyl-C(.dbd.O)--(CH.sub.2).sub.r--, wherein r is an integer in
the range from 1-10.
10. The compound according to claim 1, wherein A is
monodisperse.
11. The compound according to claim 1, wherein A is
polydisperse.
12. The compound according to claim 1, wherein the branching
chemical group B is selected from ##STR26## wherein a, b, c, d, e,
f, g, h, i are integers independently selected from the range from
0 to 24.
13. The compound according to claim 1, wherein the branching group
B is ##STR27## wherein a, b, c are integers independently selected
from the range from 0 to 24.
14. The compound according to claim 12, wherein the branching
chemical group B is selected from ##STR28## wherein a, b, c, d, e,
f, g, h, i are integers independently selected from the range from
0 to 24.
15. The compound according to claim 12, wherein the insulinotropic
agent is attached to B via the left hand terminal of B.
16. The compound according to claim 12, wherein a+b is less than 6
or a+b+c is less than 14 or a+b+c+d+e+f+g+h+l is less than 16.
17. The compound according to claim 12, wherein a is 0 or 1 and b,
c, d, e, f, h and i are all in the range from 0 to 5.
18. The compound according to claim 12, wherein a, c, d, e, g and i
are all 0 and b, f and h are all in the range from 1 to 4.
19. The compound according to claim 1, wherein a, c, d, e, g and l
are all 0 and b, f and h are all in the range from 1 to 4.
20. The compound according to any claim 1, wherein W and X are
independently selected from the bivalent connecting chemical groups
comprising amides: --C(O)--NR--, where R is hydrogen or
C.sub.1-6-alkyl, amine: --NR--, where R is hydrogen or
C.sub.1-6-alkyl, thioethers: --S--,
--S--(CH.sub.2).sub.2--SO.sub.2-- or ##STR29## ethers: --O--,
urethanes: --N(R.sup.1)--CO--N(R.sup.2)--, where R.sup.1 and
R.sup.2 independently is hydrogen or C.sub.1-6-alkyl, carbamates:
--O--CO--N(R)--, where R is hydrogen or C.sub.1-6-alkyl,
hydrazines: ##STR30## where R is hydrogen or C.sub.1-6-alkyl,
oximes: --O--N.dbd.C(--R)--, where R is hydrogen or
C.sub.1-6-alkyl, oxazolidines or thiazolidines: ##STR31##
21. The compound according to claim 20, wherein W is: --C(O)--NR--,
where R is hydrogen or C.sub.1-6-alkyl.
22. The compound according to claim 21, wherein the insulinotropic
agent is attached to W via the left hand terminal (the carbon) of
W.
23. The compound according to claim 21, wherein the insulinotropic
agent is attached to W via the right hand terminal (the nitrogen)
of W.
24. The compound according to claim 1, wherein f is 0.
25. The compound according to claim 1, wherein C* is
--(CH.sub.2).sub.n1O[(CH.sub.2).sub.n2O].sub.n3(CH.sub.2).sub.n4--,
where n1, n2 and n4 independently is an integer in the range from 1
to 10, n3 is an integer in the range from 1 to 5000, and where n3
multiplied by n2+1 is less than 10000.
26. The compound according to claim 25, wherein n2 is 2 or 3.
27. The compound according to claim 25, wherein n3 is in the range
from 1-20.
28. The compound according to claim 1, wherein C* is
--(CH.sub.2).sub.n5--, where n5 is an integer in the range from 1
to 10.
29. The compound according to claim 1, wherein Y is selected from
the bi-valent connecting chemical groups comprising amides:
--C(O)--NR--, where R is hydrogen or C.sub.1-6-alkyl, amine:
--NR--, where R is hydrogen or C.sub.1-6-alkyl, thioethers: --S--,
--S--(CH.sub.2).sub.2--SO.sub.2-- or ##STR32## ethers: --O--,
urethanes: --N(R.sup.1)--CO--N(R.sup.2)--, where R.sup.1 and
R.sup.2 independently is hydrogen or C.sub.1-6-alkyl, carbamates:
--O--CO--N(R)--, where R is hydrogen or C.sub.1-6-alkyl,
hydrazines: ##STR33## where R is hydrogen or C.sub.1-6-alkyl,
oximes: --O--N.dbd.C(--R)--, where R is hydrogen or
C.sub.1-6-alkyl, oxazolidines or thiazolidines: ##STR34##
30. The compound according to claim 1, wherein said insulinotropic
agent is a DPPIV protected peptide.
31. The compound according to claim 1, wherein said insulinotropic
agent has an EC.sub.50 of less than 1 nM as determined by the
functional receptor assay disclosed herein.
32. The compound according to claim 1, wherein said insulinotropic
agent has an EC.sub.50 of less than 300 pM, less than 200 pM or
less than 100 pM as determined by the functional receptor assay
disclosed herein.
33. The compound according to claim 1, wherein said insulinotropic
agent is derived from a peptide having a length between 27 and 45
amino acid residues in which 22 out of the first 28 amino acid
residues are identical to those found in corresponding positions in
GLP-1(7-37) (SEQ ID No. 1) or in corresponding positions in
Exendin-4(1-39) (SEQ ID No. 2).
34. The compound according to claim 1, wherein said insulinotropic
agent is derived from a peptide having a length between 28 and 45
amino acid residues in which 22 out of the first 28 amino acid
residues are identical to those found in corresponding positions in
GLP-1(7-37) or in corresponding positions in Exendin-4(1-39).
35. The compound according to claim 1, wherein said insulinotropic
agent is selected from a peptide comprising the amino acid sequence
of the formula (II):
Xaa.sub.7-Xaa.sub.8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Xaa.sub.16-Ser-Xaa.sub.18-
-Xaa.sub.19-Xaa.sub.20-Glu-Xaa.sub.22-Xaa.sub.23-Ala-Xaa.sub.25-Xaa.sub.26-
-Xaa.sub.27-Phe-Ile-Xaa.sub.30-Trp-Leu-Xaa.sub.33-Xaa.sub.34-Xaa.sub.35-Xa-
a.sub.36-Xaa.sub.37-Xaa.sub.38-Xaa.sub.39-Xaa.sub.40-Xaa.sub.41-Xaa.sub.42-
-Xaa.sub.43-Xaa.sub.44-Xaa.sub.45-Xaa.sub.46 Formula (II) (SEQ ID
No: 3) wherein Xaa.sub.7 is L-histidine, D-histidine,
desamino-histidine, 2-amino-histidine, .beta.-hydroxy-histidine,
homohistidine, N.sup..alpha.-acetyl-histidine,
.alpha.-fluoromethyl-histidine, .alpha.-methyl-histidine,
3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine; Xaa.sub.8
is Ala, D-Ala, Gly, Val, Leu, Ile, Lys, Aib,
(1-aminocyclopropyl)carboxylic acid, (1-aminocyclobutyl)carboxylic
acid, (1-aminocyclopentyl)carboxylic acid,
(1-aminocyclohexyl)carboxylic acid, (1-aminocycloheptyl)carboxylic
acid, or (1-aminocyclooctyl)carboxylic acid; Xaa.sub.16 is Val or
Leu; Xaa.sub.18 is Ser, Lys or Arg; Xaa.sub.19 is Tyr or Gln;
Xaa.sub.20 is Leu or Met; Xaa.sub.22 is Gly, Glu or Aib; Xaa.sub.23
is Gln, Glu, Lys or Arg; Xaa.sub.25 is Ala or Val; Xaa.sub.26 is
Lys, Glu or Arg; Xaa.sub.27 is Glu or Leu; Xaa.sub.30 is Ala, Glu
or Arg; Xaa.sub.33 is Val or Lys; Xaa.sub.34 is Lys, Glu, Asn or
Arg; Xaa.sub.35 is Gly or Aib; Xaa.sub.36 is Arg, Gly or Lys;
Xaa.sub.37 is Gly, Ala, Glu, Pro, Lys, amide or is absent;
Xaa.sub.38 is Lys, Ser, amide or is absent. Xaa.sub.39 is Ser, Lys,
amide or is absent; Xaa.sub.40 is Gly, amide or is absent;
Xaa.sub.41 is Ala, amide or is absent; Xaa.sub.42 is Pro, amide or
is absent; Xaa.sub.43 is Pro, amide or is absent; Xaa.sub.44 is
Pro, amide or is absent; Xaa.sub.45 is Ser, amide or is absent;
Xaa.sub.46 is amide or is absent; provided that if Xaa.sub.38,
Xaa.sub.39, Xaa.sub.40, Xaa.sub.41, Xaa.sub.42, Xaa.sub.43,
Xaa.sub.44, Xaa.sub.45 or Xaa.sub.46 is absent then each amino acid
residue downstream is also absent.
36. The compound according to claim 1, wherein said insulinotropic
agent is a peptide comprising the amino acid sequence of formula
(III):
Xaa.sub.7-Xaa.sub.8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Xaa.sub.18-Tyr-Le-
u-Glu-Xaa.sub.22-Xaa.sub.23-Ala-Ala-Xaa.sub.26-Glu-Phe-Ile-Xaa.sub.30-Trp--
Leu-Val-Xaa.sub.34-Xaa.sub.35-Xaa.sub.36-Xaa.sub.37-Xaa.sub.38
Formula (III) (SEQ ID No: 4) wherein Xaa.sub.7 is L-histidine,
D-histidine, desamino-histidine, 2-amino-histidine,
.beta.-hydroxy-histidine, homohistidine,
N.sup..alpha.-acetyl-histidine, .alpha.-fluoromethyl-histidine,
.alpha.-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or
4-pyridylalanine; Xaa.sub.8 is Ala, D-Ala, Gly, Val, Leu, Ile, Lys,
Aib, (1-aminocyclopropyl)carboxylic acid,
(1-aminocyclobutyl)carboxylic acid, (1-aminocyclopentyl)carboxylic
acid, (1-aminocyclohexyl)carboxylic acid,
(1-aminocycloheptyl)carboxylic acid, or
(1-aminocyclooctyl)carboxylic acid; Xaa.sub.18 is Ser, Lys or Arg;
Xaa.sub.22 is Gly, Glu or Aib; Xaa.sub.23 is Gln, Glu, Lys or Arg;
Xaa.sub.26 is Lys, Glu or Arg; Xaa.sub.30 is Ala, Glu or Arg;
Xaa.sub.34 is Lys, Glu or Arg; Xaa.sub.35 is Gly or Aib; Xaa.sub.36
is Arg or Lys; Xaa.sub.37 is Gly, Ala, Glu or Lys; Xaa.sub.38 is
Lys, NH.sub.2 or is absent.
37. The compound according to claim 1, wherein said insulinotropic
agent is selected from GLP-1(7-35), GLP-1(7-36), GLP-1(7-36)-amide,
GLP-1(7-37), GLP-1(7-38), GLP-1(7-39), GLP-1(7-40), GLP-1(7-41) or
an analogue thereof.
38. The compound according to claim 1, wherein said insulinotropic
agent comprises no more than fifteen amino acid residues which have
been exchanged, added or deleted as compared to GLP-1(7-37) (SEQ ID
No. 1), or no more than ten amino acid residues which have been
exchanged, added or deleted as compared to GLP-1(7-37) (SEQ ID No.
1).
39. The compound according to claim 38, wherein said insulinotropic
agent comprises no more than six amino acid residues which have
been exchanged, added or deleted as compared to GLP-1(7-37) (SEQ ID
No. 1).
40. The compound according to claim 1, wherein said insulinotropic
agent comprises no more than 4 amino acid residues which are not
encoded by the genetic code.
41. The compound according to claim 1, wherein said insulinotropic
agent comprises an Aib residue as the second amino acid residue
from the N-terminal.
42. The compound according to claim 1, wherein the N-terminal amino
acid residue (position 7 in formulae II and III) of said
insulinotropic agent is selected from the group consisting of
D-histidine, desamino-histidine, 2-amino-histidine,
.beta.-hydroxy-histidine, homohistidine,
N.sup..alpha.-acetyl-histidine, .alpha.-fluoromethyl-histidine,
.alpha.-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine and
4-pyridylalanine.
43. The compound according to claim 1, wherein said insulinotropic
agent is selected from the group consisting of
[Arg.sup.34]GLP-1(7-37), [Arg.sup.26,34]GLP-1(7-37)Lys,
[Lys.sup.36Arg.sup.26,34]GLP-1(7-36), [Aib.sup.8,22,35]GLP-1(7-37),
[Aib.sup.8,35]GLP-1(7-37), [Aib.sup.8,22]GLP-1(7-37),
[Aib.sup.8,22,35 Arg.sup.26,34]GLP-1(7-37)Lys,
[Aib.sup.8,35Arg.sup.26,34]GLP-1(7-37)Lys,
[Aib.sup.8,22Arg.sup.26,34]GLP-1(7-37)Lys,
[Aib.sup.8,22,35Arg.sup.26,34]GLP-1(7-37) Lys,
[Aib.sup.8,35Arg.sup.26,34]GLP-1(7-37)Lys,
[Aib.sup.8,22,35Arg.sup.26]GLP-1(7-37) Lys,
[Aib.sup.8,35Arg.sup.26]GLP-1(7-37)Lys,
[Aib.sup.8,22Arg.sup.26]GLP-1(7-37)Lys,
[Aib.sup.8,22,35Arg.sup.34]GLP-1(7-37)Lys,
[Aib.sup.8,35Arg.sup.34]GLP-1(7-37)Lys,
[Aib.sup.8,22Arg.sup.34]GLP-1(7-37) Lys,
[Aib.sup.8,22,35Ala.sup.37]GLP-1(7-37)Lys,
[Aib.sup.8,35Ala.sup.37]GLP-1(7-37) Lys,
[Aib.sup.8,22Ala.sup.37]GLP-1(7-37)Lys,
[Aib.sup.8,22,35Lys.sup.37]GLP-1(7-37),
[Aib.sup.8,35Lys.sup.37]GLP-1(7-37),
[Aib.sup.8,22Lys.sup.37]GLP-1(7-37) or derivatives thereof which
has been amidated on the C-terminal.
44. The compound according to claim 1, wherein said insulinotropic
agent comprises at least one Aib residue.
45. The compound according to claim 1, wherein said insulinotropic
agent contains two Aib residues.
46. The compound according to claim 1, wherein said insulinotropic
agent comprises a serine residue at position 18 relative to
GLP-1(7-37) (SEQ ID. No. 1), corresponding to position 12 relative
to Exendin-4(1-39).
47. The compound according to any one of the previous claims,
wherein said insulinotropic agent comprises a tyrosine residue at
position 19 relative to GLP-1(7-37) (SEQ ID. No. 1), corresponding
to position 13 relative to Exendin-4(1-39).
48. The compound according to claim 1, wherein said insulinotropic
agent comprises a glycine residue at position 22 relative to
GLP-1(7-37) (SEQ ID. No. 1), corresponding to position 16 relative
to Exendin-4(1-39).
49. The compound according to claim 1, wherein said insulinotropic
agent comprises a glutamine residue at position 23 relative to
GLP-1(7-37) (SEQ ID. No. 1), corresponding to position 17 relative
to Exendin-4(1-39).
50. The compound according to claim 1, wherein said insulinotropic
agent comprises a lysine residue at position 26 relative to
GLP-1(7-37) (SEQ ID. No. 1), corresponding to position 20 relative
to Exendin-4(1-39).
51. The compound according to claim 1, wherein said insulinotropic
agent comprises a glutamate residue at position 27 relative to
GLP-1(7-37) (SEQ ID. No. 1), corresponding to position 21 relative
to Exendin-4(1-39).
52. The compound according to claim 1, wherein said insulinotropic
agent is exendin-4(1-39).
53. The compound according to claim 1, wherein said insulinotropic
agent is [Ser.sup.38Lys.sup.39]Exendin-4(1-39)LysLysLysLysLys-amide
(SEQ ID No. 5).
54. The compound according to claim 1, wherein said insulinotropic
agent is attached to Y--C*-Q or Q via the amino acid residue in
position 25 to 45 relative to the amino acid sequence SEQ ID
No:1.
55. The compound according to claim 1, wherein said insulinotropic
agent is attached to Y--C*-Q or Q via an amino acid residue
selected from one of the 10 C-terminal amino acid residues.
56. The compound according to claim 1, wherein said insulinotropic
agent is attached to Y--C*-Q or Q via the amino acid residue in
position 23, 26, 34, 36 or 38 relative to the amino acid sequence
SEQ ID No:1.
57. The compound according to claim 1, wherein said insulinotropic
agent is attached to Y--C*-Q or Q via the amino acid residue in
position 17, 20, 28, 30 or 32 relative to the amino acid sequence
SEQ ID No:2.
58. The compound according to claim 1, wherein said insulinotropic
agent is attached to Y--C*-Q or Q via the C-terminal amino acid
residue.
59. The compound according to claim 1, wherein said insulinotropic
agent is attached to Y--C*-Q or Q via a carboxyl group, an amino
group, a keto group, a hydroxyl group, a thiol group or a hydrazide
group.
60. The compound according to claim 1, wherein said insulinotropic
agent is attached to Y--C*-Q or Q via a the epsilon-amino group on
a lysine residue.
61. The compound according to claim 60, wherein said insulinotropic
agent comprises only one lysine residue.
62. The compound according to claim 61, wherein said lysine residue
is the C-terminal amino acid residue of said insulinotropic
agent.
63. The compound according to claim 1, wherein said compound has an
EC.sub.50 of less than 1000 pM, less than 500 pM as determined by a
functional receptor assay.
64. The compound according to claim 1, wherein said compound is
selected from the group consisting of
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has
a molecular weight of approximately 20 kDa,
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 20 kDa,
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 5 kDa,
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 2 kDa,
N.sup..epsilon.26-(3-(mPEGyl)propionyl)[Aib.sup.8Glu.sup.22,30,Lys.sup.33-
,Asn.sup.34,Gly.sup.35,36, Pro.sup.37]GLP-1(7-37)ylSerSerGly
AlaProProProSer amide wherein mPEGyl is polydisperse and has a
molecular weight of approximately 2 kDa,
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl)Lysinamide) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 750 Da,
N.sup..epsilon.-[Aib.sup.8,22,35]GLP-1(7-37)yl(S.sup..epsilon.-(1-mPEGylp-
ropyl-2,5-dioxo-pyrrolidin-3-yl)Cysteinamide wherein mPEGyl is
polydisperse and has a molecular weight of approximately 5000 Da,
N.sup..alpha.-(3(3H-imidazol-4-yl)-propionyl
[Aib.sup.22,35,Arg.sup.26,34]GLP-1-(8-37))yl(N.sup..epsilon.-(3-(mPEGyl)p-
ropionyl)Lysinamide) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 2000 Da,
N.sup..epsilon.26-(3-(mPEGyl)propionyl)[Arg.sup.34]GLP-1-(7-37)
wherein mPGyl is polydisperse and has a molecular weight of
approximately 2 kDa, and
(S)--N--((S)-5-(N--((S)-5-carbamoyl-5-(mPEGylpropionylamino)pentyl)c-
arbamoyl)-5-(mPEGylpropionylamino)pentyl)-5-(N.sup..alpha.7-(3-(4-imidazol-
yl)propionyl)[Aib.sup.22,35,Arg.sup.26,34]GLP-1-(8-37)yl)-2-(mPEGylpropion-
ylamino)hexanoic amide wherein mPEGyl is polydisperse and has a
molecular weight of approximately 750 Da,
65. A pharmaceutical composition comprising a compound according to
claim 1 and a pharmaceutically acceptable excipient.
66. The pharmaceutical composition according to claim 65, which is
suited for pulmonary administration.
67. A method for treating hyperglycemia, type 2 diabetes, impaired
glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome
X, dyslipidemia, cognitive disorders, atheroschlerosis, myocardial
infarction, coronary heart disease and other cardiovascular
disorders, stroke, inflammatory bowel syndrome, dyspepsia and
gastric ulcers, said method comprising administering to a subject
in need of such treatment an effective amount of a pharmaceutical
composition according to claim 65.
68. A method for delaying disease progression in type 2 diabetes in
a subject, said method comprising administering to said subject an
effective amount of a pharmaceutical composition according to claim
65.
69. A method for decreasing food intake, decreasing .beta.-cell
apoptosis, increasing .beta.-cell function and .beta.-cell mass,
and/or for restoring glucose sensitivity to .beta.-cells in a
subject, said method comprising administering to said subject an
effective amount of a pharmaceutical composition according to claim
65.
Description
CROSS-REFERENCE TO RELATED PATENT APPLICATIONS
[0001] This application is a continuation International Patent
Application No. PCT/DK2004/00886 filed Dec. 17, 2004 and claims
priority of U.S. Patent Application Nos. 60/531,053, filed Dec. 19,
2003, and 60/587,181, filed Jul. 12, 2004 and Danish Patent
Application Nos. PA 2003 01885, filed Dec. 18, 2003 and PA 2004
01090, filed Jul. 9, 2004.
FIELD OF THE INVENTION
[0002] The present invention relates to novel GLP-1 compounds, to
pharmaceutical compositions comprising these compounds and to the
use of the compounds for the treatment of diseases related to
diabetes.
BACKGROUND OF THE INVENTION
[0003] Diabetes mellitus is a metabolic disorder in which the
ability to utilize glucose is partly or completely lost. About 5%
of all people suffer from diabetes and the disorder approaches
epidemic proportions. Since the introduction of insulin in the
1920's, continuous efforts have been made to improve the treatment
of diabetes mellitus.
[0004] One peptide expected to become very important in the
treatment of diabetes is glucagon-like peptide-1 (GLP-1). Human
GLP-1 is a 37 amino acid residue peptide originating from
pre-proglucagon which is synthesized i.a. in the L-cells in the
distal ileum, in the pancreas and in the brain. GLP-1 is an
important gut hormone with regulatory function in glucose
metabolism and gastrointestinal secretion and metabolism. GLP-1
stimulates insulin secretion in a glucose-dependant manner,
stimulates insulin biosynthesis, promotes beta cell rescue,
decreases glucagon secretion, gastric emptying and food intake.
Human GLP-1 is hydrolysed to GLP-1(7-37) and GLP-1(7-36)-amide
which are both insulinotropic peptides. A simple system is used to
describe fragments and analogues of this peptide. Thus, for
example, [Gly.sup.8]GLP-1(7-37) designates an analogue of
GLP-1(7-37) formally derived from GLP-1(7-37) by substituting the
naturally occurring amino acid residue in position 8 (Ala) by Gly.
Similarly, (N.sup..epsilon.34-tetradecanoyl)[Lys.sup.34]GLP-1(7-37)
designates GLP-1(7-37) wherein the .epsilon.-amino group of the Lys
residue in position 34 has been tetradecanoylated. PCT publications
WO 98/08871 and WO 99/43706 disclose stable derivatives of GLP-1
analogues, which have a lipophilic substituent. These stable
derivatives of GLP-1 analogues have a protracted profile of action
compared to the corresponding GLP-1 analogues.
[0005] In the last decade a number of peptides have been isolated
from the venom of the Gila monster lizards (Heloderma suspectum and
Heloderma horridum). Exendin-4 is a 39 amino acid residue peptide
isolated from the venom of Heloderma suspectum, and this peptide
shares 52% homology with GLP-1(7-37) in the overlapping region.
Exendin-4 is a potent GLP-1 receptor agonist which has been shown
to stimulate insulin release and ensuing lowering of the blood
glucose level when injected into dogs. The group of
exendin-4(1-39), certain fragments thereof, analogs thereof and
derivatives thereof, are potent insulinotropic agents. Most
importantly the group of exendin-4(1-39), insulinotropic fragments
thereof, insulinotropic analogs thereof and insulinotropic
derivatives thereof.
[0006] Common to GLP-1 and exendins are that an extensive amount of
variants have been synthesized and studied in particular in
relation the plasma half-life. Low plasma half-lifes may be due to
chemical stability towards peptidases (mainly dipeptidyl
aminopeptidase IV) and to renal clearance. However, these analogues
and derivatives of insulionotropic peptides lack a satisfactory
bioavailability when administered by the pulmonary route, i.e. when
administered to the lower respirary tract such as through the
bronchioles or alveoli.
[0007] WO 00/66629 discloses modified exendin agonists which have
been coupled to polyethyleneglycol via a lysine residue to decrease
renal clearance.
[0008] WO 03/40309 discloses peptide acting as both GLP-1 receptor
agonists and glucagon receptor antagonists. Among the disclosed
peptides are two peptides which have been coupled to
polyethyleneglycol via a C-terminal cycteine residue.
[0009] WO 2004/093823 discloses polyethylene glycolated GLOP-1
peptides.
[0010] Pulmonary administration of GLP-1 peptides have been
disclosed in WO 01/51071 and WO 00/12116.
[0011] The insulinotropic peptides derived from GLP-1 and Exendin-4
stimulatesd insulin release only when plasma glucose levels are
high, the risk of hypoglycaemic events is reduced. Thus, the
peptides are particularly useful for patients with diabetes who no
longer respond to OHA's (oral hyperglycaemic agents) and who should
from a strict medical point of view be administered insulin.
Patients and to some extent also doctors are often not keen on
initiating insulin treatment before this is absolutely necessary,
presumably because of the fear of hypoglycaemic events or the fear
of injections/needles. Thus, there is a need for insulinotropic
peptides which are sufficiently potent and which can be
administered by the pulmonary route. Thus, it is an object of the
present invention to provide insulinotropic peptides which have
sufficient pulmonary bioavailability to serve as an alternative to
peptides for paranteral administration. Insulinotropic peptides
having pulmonary bioavailability is a balance between potency and
bioavailability. It is also an object of the present invention to
provide insulinotropic peptides which are less prone to
aggregation, a well known problem associated with the glucagon-like
peptides. Being less prone to aggregation facilitates economical
manufacturing processes as well as enabling the compounds to be
administered by medical infusion pumps.
Definitions
[0012] In the present specification, the following terms have the
indicated meaning:
[0013] The term "polypeptide" and "peptide" as used herein means a
compound composed of at least five constituent amino acids
connected by peptide bonds. The constituent amino acids may be from
the group of the amino acids encoded by the genetic code and they
may natural amino acids which are not encoded by the genetic code,
as well as synthetic amino acids. Natural amino acids which are not
encoded by the genetic code are e.g. hydroxyproline,
.gamma.-carboxyglutamate, ornithine, phosphoserine, D-alanine and
D-glutamine. Synthetic amino acids comprise amino acids
manufactured by chemical synthesis, i.e. D-isomers of the amino
acids encoded by the genetic code such as D-alanine and D-leucine,
Aib (.alpha.-aminoisobutyric acid), Abu (.alpha.-aminobutyric
acid), Tle (tert-butylglycine), .beta.-alanine, 3-aminomethyl
benzoic acid, anthranilic acid.
[0014] The term "analogue" as used herein referring to a
polypeptide means a modified peptide wherein one or more amino acid
residues of the peptide have been substituted by other amino acid
residues and/or wherein one or more amino acid residues have been
deleted from the peptide and/or wherein one or more amino acid
residues have been deleted from the peptide and or wherein one or
more amino acid residues have been added to the peptide. Such
addition or deletion of amino acid residues can take place at the
N-terminal of the peptide and/or at the C-terminal of the peptide.
A simple system is often used to describe analogues: For example
[Arg.sup.34]GLP-1(7-37)Lys designates a GLP-1(7-37) analogue
wherein the naturally occuring lysine at position 34 has been
substituted with arginine and wherein a lysine has been added to
the terminal amino acid residue, i.e. to the Gly.sup.37. All amino
acids for which the optical isomer is not stated is to be
understood to mean the L-isomer.
[0015] The term "derivative" as used herein in relation to a
peptide means a chemically modified peptide or an analogue thereof,
wherein at least one substituent is not present in the unmodified
peptide or an analogue thereof, i.e. a peptide which has been
covalently modified. Typical modifications are amides,
carbohydrates, alkyl groups, acyl groups, esters and the like. An
example of a derivative of GLP-1(7-37) is
N.sup..epsilon.26-((4S)-4-(hexadecanoylamino)-butanoyl)[Arg.sup.34,Lys.su-
p.26]GLP-1-(7-37).
[0016] The term "insulinotropic agent" as used herein means a
compound which is an agonist of the human GLP-1 receptor, i.e. a
compound which stimulates the formation of cAMP in a suitable
medium containing the human GLP-1 receptor (one such medium
disclosed below). The potency of an insulinotropic agent is
determined by calculating the EC.sub.50 value from the
dose-response curve as described below.
[0017] Baby hamster kidney (BHK) cells expressing the cloned human
GLP-1 receptor (BHK-467-12A) were grown in DMEM media with the
addition of 100 IU/mL penicillin, 100 .mu.g/mL streptomycin, 5%
fetal calf serum and 0.5 mg/mL Geneticin G-418 (Life Technologies).
The cells were washed twice in phosphate buffered saline and
harvested with Versene. Plasma membranes were prepared from the
cells by homogenisation with an Ultraturrax in buffer 1 (20 mM
HEPES-Na, 10 mM EDTA, pH 7.4). The homogenate was centrifuged at
48,000.times.g for 15 min at 4.degree. C. The pellet was suspended
by homogenization in buffer 2 (20 mM HEPES-Na, 0.1 mM EDTA, pH
7.4), then centrifuged at 48,000.times.g for 15 min at 4.degree. C.
The washing procedure was repeated one more time. The final pellet
was suspended in buffer 2 and used immediately for assays or stored
at -80.degree. C.
[0018] The functional receptor assay was carried out by measuring
cyclic AMP (cAMP) as a response to stimulation by the
insulinotropic agent. cAMP formed was quantified by the
AlphaScreen.TM. cAMP Kit (Perkin Elmer Life Sciences). Incubations
were carried out in half-area 96-well microtiter plates in a total
volume of 50 .mu.L buffer 3 (50 mM Tris-HCl, 5 mM HEPES, 10 mM
MgCl.sub.2, pH 7.4) and with the following addiditions: 1 mM ATP, 1
.mu.M GTP, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 0.01%
Tween-20, 0.1% BSA, 6 .mu.g membrane preparation, 15 .mu.g/mL
acceptor beads, 20 .mu.g/mL donor beads preincubated with 6 nM
biotinyl-cAMP. Compounds to be tested for agonist activity were
dissolved and diluted in buffer 3. GTP was freshly prepared for
each experiment. The plate was incubated in the dark with slow
agitation for three hours at room temperature followed by counting
in the Fusion.TM. instrument (Perkin Elmer Life Sciences).
Concentration-response curves were plotted for the individual
compounds and EC.sub.50 values estimated using a four-parameter
logistic model with Prism v. 4.0 (GraphPad, Carlsbad, Calif.).
[0019] The term "GLP-1 peptide" as used herein means GLP-1(7-37)
(SEQ ID No 1), a GLP-1 (7-37) analogue, a GLP-1(7-37) derivative or
a derivative of a GLP-1(7-37) analogue. In one embodiment the GLP-1
peptide is an insulinotropic agent.
[0020] The term "exendin-4 peptide" as used herein means
exendin-4(1-39) (SEQ ID No 2), an exendin-4(1-39) analogue, an
exendin-4(1-39) derivative or a derivative of an exendin-4(1-39)
analogue. In one embodiment the exendin-4 peptide is an
insulinotropic agent.
[0021] The term "DPP-IV protected" as used herein referring to a
polypeptide means a polypeptide which has been chemically modified
in order to render said compound resistant to the plasma peptidase
dipeptidyl aminopeptidase-4 (DPP-IV). The DPP-IV enzyme in plasma
is known to be involved in the degradation of several peptide
hormones, e.g. GLP-1, GLP-2, Exendin-4 etc. Thus, a considerable
effort is being made to develop analogues and derivatives of the
polypeptides susceptible to DPP-IV mediated hydrolysis in order to
reduce the rate of degradation by DPP-IV. In one embodiment a
DPP-IV protected peptide is more resistant to DPP-IV than
GLP-1(7-37) or Exendin-4(1-39).
Resistance of a peptide to degradation by dipeptidyl aminopeptidase
IV is determined by the following degradation assay:
[0022] Aliquots of the peptide (5 nmol) are incubated at 37.degree.
C. with 1 .mu.L of purified dipeptidyl aminopeptidase IV
corresponding to an enzymatic activity of 5 mU for 10-180 minutes
in 100 .mu.L of 0.1 M triethylamine-HCl buffer, pH 7.4. Enzymatic
reactions are terminated by the addition of 5 .mu.L of 10%
trifluoroacetic acid, and the peptide degradation products are
separated and quantified using HPLC analysis. One method for
performing this analysis is: The mixtures are applied onto a Vydac
C18 widepore (30 nm pores, 5 .mu.m particles) 250.times.4.6 mm
column and eluted at a flow rate of 1 ml/min with linear stepwise
gradients of acetonitrile in 0.1% trifluoroacetic acid (0%
acetonitrile for 3 min, 0-24% acetonitrile for 17 min, 24-48%
acetonitrile for 1 min) according to Siegel et al., Regul. Pept.
1999; 79:93-102 and Mentlein et al. Eur. J. Biochem. 1993;
214:829-35. Peptides and their degradation products may be
monitored by their absorbance at 220 nm (peptide bonds) or 280 nm
(aromatic amino acids), and are quantified by integration of their
peak areas related to those of standards. The rate of hydrolysis of
a peptide by dipeptidyl aminopeptidase IV is estimated at
incubation times which result in less than 10% of the peptide being
hydrolysed.
[0023] The term "mPEGyl" means a polydisperse or monodisperse
radical of the structure ##STR1## wherein m is an integer larger
than 1. Thus, a mPEGyl wherein m is 90 has a molecular weight of
3991 Da, i.e. approx 4 kDa. Likewise, a mPEGyl with an average
molecular weigt of 20 kDa has an average m of 454. Due to the
process for producing mPEGyl these molecues often have a
distribution of molecular weights. This distribution is described
by the polydispersity index.
[0024] The term "polydispersity index" as used herein means the
ratio between the weight average molecular weight and the number
average molecular weight, as known in the art of polymer chemistry
(see e.g. "Polymer Synthesis and Characterization", J.a: Nairn,
Uiversity of Utah, 2003). The polydispersity index is a number
which is greater than or qual to one, and it may be estimated from
Gel Permeation Chromatographic data. When the polydispersity index
is one the product is monodisperse, and is thus made up of a single
moleculer weight. When the polydispersity index is greater than one
it is a measure of the polydispersity of that polymer, i.e. how
broad the distribution of polymers with different molecular weights
is.
[0025] The term "C.sub.1-6-alkyl" as used herein means a saturated,
branched, straight or cyclic hydrocarbon group having from 1 to 6
carbon atoms. Representative examples include, but are not limited
to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl,
tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl,
isohexyl, cyclohexane and the like.
[0026] he term "pharmaceutically acceptable" as used herein means
suited for normal pharmaceutical applications, i.e. giving rise to
no adverse events in patients etc.
[0027] The term "heavy atom" as used herein means an atom having a
molar weight equal to or larger than carbon, e.g. C, N, O and
S.
[0028] The term "excipient" as used herein means the chemical
compounds which are normally added to pharmaceutical compositions,
e.g. buffers, tonicity agents, preservatives and the like.
[0029] The term "effective amount" as used herein means a dosage
which is sufficient to be effective for the treatment of the
patient compared with no treatment.
[0030] The term "pharmaceutical composition" as used herein means a
product comprising an active compound or a salt thereof together
with pharmaceutical excipients such as buffer, preservative, and
optionally a tonicity modifier and/or a stabilizer. Thus a
pharmaceutical composition is also known in the art as a
pharmaceutical formulation.
[0031] The term "treatment of a disease" as used herein means the
management and care of a patient having developed the disease,
condition or disorder. The purpose of treatment is to combat the
disease, condition or disorder. Treatment includes the
administration of the active compounds to eliminate or control the
disease, condition or disorder as well as to alleviate the symptoms
or complications associated with the disease, condition or
disorder.
DESCRIPTION OF THE INVENTION
[0032] In one aspect the present invention relates to a compound
having the structure of the formula (I): Insulinotropic
agent(--Y--C*).sub.f-Q (I) wherein Insulinotropic agent is a
radical derived from an insulinotropic peptide which binds to the
human GLP-1 receptor, or a redical derived from a peptide in which
22 positions out of the first 30 are identical to those found in
corresponding positions in GLP-1 or found in corresponding
positions in Exendin-4, and Y is a bivalent connecting chemical
group connecting C* with the Insulinotropic agent, and C* is a
bivalent polar separating chemical group where 50-20% of the heavy
atoms are either O or N, and f is 0 or 1 and Q is selected from
##STR2## wherein
[0033] A is a polar chemical group of a single molecular size
(monodisperse) or of several molecular sizes (polydisperse)
where
[0034] 50-20% of the heavy atoms are independently oxygen or
nitrogen, and
[0035] W is a bivalent chemical group whereby A is connected,
and
[0036] X is a bivalent connecting chemical group whereby B is
connected, and
[0037] B is a connecting or branching chemical group.
In another aspect the present invention relates to a compound
having the structure of the formula (I): Insulinotropic
agent(--Y--C*).sub.f-Q (I) wherein Insulinotropic agent is a
redical derived from an insulinotropic peptide which binds to the
human GLP-1 receptor, or a radical derived from a peptide in which
22 positions out of the first 30 are identical to those found in
corresponding positions in GLP-1 or found in corresponding
positions in Exendin-4, with the proviso that the C-terminal amino
acid residue of said insulinotropic agent is not cysteine, and Y is
a bivalent connecting chemical group connecting C* with the
Insulinotropic agent, and C* is a bivalent polar separating
chemical group where 50-20% of the heavy atoms are either O or N,
and f is 0 or 1 and Q is selected from ##STR3## wherein
[0038] A is a polar chemical group of a single molecular size
(monodisperse) or of several molecular sizes (polydisperse)
where
[0039] 50-20% of the heavy atoms are independently oxygen or
nitrogen, and
[0040] W is a bivalent chemical group whereby A is connected,
and
[0041] X is a bivalent connecting chemical group whereby B is
connected, and
[0042] B is a connecting or branching chemical group.
[0043] The general formula (I) and the encompassed peptide radical
is to be understood as follows.
The following compound is encompassed by formula (I):
[0044]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37-
]GLP-1(7-37) amide wherein mPEGyl is polydisperse and has a
molecular weight of approximately 2 kDa ##STR4##
[0045] The insulinotropic agent is the radical comprising the
peptide including the four methylene groups in the lysine residue
in position 37. The group A is the mPEGyl-CH.sub.2CH.sub.2--
wherein mPEGyl has a molecular weight of approximately 2 kDa. The
bivalent chemical group W whereby mPEGyl-CH.sub.2CH.sub.2-- is
connected to the radical derived from the insulinotropic peptide is
the amide --C(O)--NH--.
[0046] In one embodiment of the invention A is a monodisperse or
polydisperse chemical group having the structure
--(CH.sub.2).sub.lO[(CH.sub.2).sub.nO].sub.m(CH.sub.2).sub.p--H,
where l, n and p independently are an integer in the range from 1
to 10, m is an integer in the range from 1 to 5000, and where m
multiplied by n+1 is less than 10000.
[0047] In another embodiment of the invention A is a monodisperse
or polydisperse chemical group having the structure
--(CH.sub.2).sub.lC(.dbd.O)O[(CH.sub.2).sub.nO].sub.m(CH.sub.2).sub.p--H,
where l, n and p independently are an integer in the range from 1
to 10, m is an integer in the range from 1 to 5000, and where m
multiplied by n+1 is less than 10000.
[0048] In another embodiment of the invention n is 2 or 3.
[0049] In another embodiment of the invention m is in the range
from 10-1000, or in the range from 20-250.
[0050] In another embodiment of the invention A is a monodisperse
or polydisperse chemical group having the structure
-(Z.sup.1(CH.sub.2).sub.fO[(CH.sub.2).sub.2O].sub.m(CH.sub.2).sub.p--NR.s-
up.1).sub.q-Z.sup.2, where Z.sup.1 is --CO-- or
--CO--(CH.sub.2).sub.n--CO--NH--, and Z.sup.2 is --R.sup.1,
--CO--(CH.sub.2).sub.n--R.sup.1,
--(CH.sub.2).sub.lO[(CH.sub.2).sub.2O].sub.m(CH.sub.2).sub.p--R.sup.1
wherein l and n and p independently are integers in the range from
1 to 10, and R.sup.1 is --OH, --NH.sub.2, --NH--R.sup.2,
--NH(--R.sup.2)--R.sup.2, --COOH, C.sub.1-6-alkyl, or
--NH--CH(R.sup.2)--COOH, and where m and q are independently
integers in the range from 1 to 20, and where l, n and p are
independently integers in the range from 1 to 6, and R.sup.2 is
hydrogen or C.sub.1-6-alkyl.
[0051] In another embodiment of the invention A is mPEGyl.
[0052] In another embodiment of the invention A is
mPEGyl-C(.dbd.O)--(CH.sub.2).sub.r--, wherein r is an integer in
the range from 1-10.
[0053] In another embodiment of the invention A is monodisperse,
i.e. it is made up of only one component.
[0054] In another embodiment of the invention A has a
polydispersity index from 1.00 to 1.10.
[0055] In another embodiment of the invention A is polydisperse and
preferably having a polydispersity index which is less than 1.2,
less than 1.1, less than 1.05, less than 1.03, less than 1.02, less
than 1.010, less than 1.008, less than 1.005 or less than
1.0025.
[0056] In another embodiment of the invention the branching
chemical group B is selected from ##STR5## wherein a, b, c, d, e,
f, g, h, i are integers independently selected from the range from
0 to 24. In another embodiment of the invention the branching group
B is ##STR6## wherein a, b, c are integers independently selected
from the range from 0 to 24. In another embodiment of the invention
the branching chemical group B is selected from ##STR7## wherein a,
b, c, d, e, f, g, h, i are integers independently selected from the
range from 0 to 24.
[0057] In another embodiment of the invention the insulinotropic
agent is attached to B via the left hand terminal of B.
[0058] In another embodiment of the invention a+b is less than 6 or
a+b+c is less than 14 or a+b+c+d+e+f+g+h+l is less than 16.
[0059] In another embodiment of the invention a is 0 or 1 and b, c,
d, e, f, h and i are all in the range from 0 to 5.
[0060] In another embodiment of the invention a, c, d, e, g and i
are all 0 and b, f and h are all in the range from 1 to 4.
[0061] In another embodiment of the invention a, c, d, e, g and l
are all 0 and b, f and h are all in the range from 1 to 4.
[0062] In another embodiment of the invention, W and X are
independently selected from the bi-valent connecting chemical
groups comprising
amides: --C(O)--NR--, where R is hydrogen or C.sub.1-6-alkyl,
amines: --NR--, where R is hydrogen or C.sub.1-6-alkyl,
[0063] thioethers: --S--, --S--(CH.sub.2).sub.2--SO.sub.2-- or
##STR8## ethers: --O--, urethanes: --N(R.sup.1)--CO--N(R.sup.2)--,
where R.sup.1 and R.sup.2 independently is hydrogen or
C.sub.1-6-alkyl, carbamates: --O--CO--N(R)--, where R is hydrogen
or C.sub.1-6-alkyl, hydrazines: ##STR9## where R is hydrogen or
C.sub.1-6-alkyl, oximes: --O--N.dbd.C(--R)--, where R is hydrogen
or C.sub.1-6-alkyl, oxazolidines or thiazolidines: ##STR10##
[0064] The hydrazine derivatives of the formula, ##STR11## where R
is hydrogen or C.sub.1-6-alkyl may be formed by reaction of an
aldehyde derivative (--CO--H) or a ketone derivative (--CO--R)
with
[0065] hydrazine derivatives (--NH--NH.sub.2) or
[0066] hydrazine carboxylate derivatives (--O--C(O)--NH--NH.sub.2)
or
[0067] semicarbazide derivatives (--NH--C(O)--NH--NH.sub.2) or
[0068] thiosemicarbazide derivatives (--NH--C(S)--NH--NH.sub.2)
or
[0069] carbonic acid dihydrazide derivatives
(--NHC(O)--NH--NH--C(O)--NH--NH.sub.2) or
[0070] carbazide derivatives (--NH--NH--C(O)--NH--NH.sub.2) or
[0071] thiocarbazide derivatives (--NH--NH--C(S)--NH--NH.sub.2)
or
[0072] aryl hydrazide derivatives
(--NH--C(O)--C.sub.6H.sub.4--NH--NH.sub.2) or
[0073] hydrazide derivatives (--C(O)--NH--NH.sub.2).
The oximes of the formula --O--N.dbd.C(--R)--, where R is hydrogen
or C.sub.1-6-alkyl and may be formed by reaction of an aldehyde
(--CO--H) or a ketone (--CO--R) with
[0074] oxylamine (--O--NH.sub.2) or
[0075] --C(O)--O--NH.sub.2 or
[0076] --NH--C(O)--O--NH.sub.2 or
[0077] --NH--C(S)--O--NH.sub.2.
[0078] In another embodiment of the invention W is --C(O)--NR--,
where R is hydrogen or C.sub.1-6-alkyl.
[0079] In another embodiment of the invention the insulinotropic
agent is attached to W via the left hand terminal (the carbon) of
W.
[0080] In another embodiment of the invention the insulinotropic
agent is attached to W via the right hand terminal (the nitrogen)
of W.
[0081] In another embodiment of the invention, f is 0.
[0082] In another embodiment of the invention C* is
--(CH.sub.2).sub.n1O[(CH.sub.2).sub.n2O].sub.n3(CH.sub.2).sub.n4--,
where n1, n2 and n4 independently is an integer in the range from 1
to 10, n3 is an integer in the range from 1 to 5000, and where n3
multiplied by n2+1 is less than 10000.
[0083] In another embodiment of the invention n2 is 2 or 3.
[0084] In another embodiment of the invention n3 is in the range
from 1-20.
[0085] In another embodiment of the invention C* is
--(CH.sub.2).sub.n5--, where n5 is an integer in the range from 1
to 10.
[0086] In another embodiment of the invention Y is selected from
the bi-valent connecting chemical groups comprising
amides: --C(O)--NR--, where R is hydrogen or C.sub.1-6-alkyl,
amines: --NR--, where R is hydrogen or C.sub.1-6-alkyl,
[0087] thioethers: --S--, --S--(CH.sub.2).sub.2--SO.sub.2-- or
##STR12## ethers: --O--, urethanes: --N(R.sup.1)--CO--N(R.sup.2)--,
where R.sup.1 and R.sup.2 independently is hydrogen or
C.sub.1-6-alkyl, carbamates: --O--CO--N(R)--, where R is hydrogen
or C.sub.1-6-alkyl, hydrazines: ##STR13## where R is hydrogen or
C.sub.1-6-alkyl, oximes: --O--N.dbd.C(--R)--, where R is hydrogen
or C.sub.1-6-alkyl, oxazolidines or thiazolidines: ##STR14##
[0088] In another embodiment of the invention the insulinotropic
agent is a DPPIV protected peptide.
[0089] In another embodiment of the invention the insulinotropic
agent has an EC.sub.50 of less than 1 nM as determined by the
functional receptor assay disclosed herein.
[0090] In another embodiment of the invention the insulinotropic
agent has an EC.sub.50 of less than 300 pM, less than 200 pM or
less than 100 pM as determined by the functional receptor assay
disclosed herein.
[0091] In another embodiment of the invention the insulinotropic
agent is derived from a peptide having a length between 27 and 45
amino acid residues in which 22 out of the first 28 amino acid
residues are identical to those found in corresponding positions in
GLP-1(7-37) (SEQ ID No. 1) or in corresponding positions in
Exendin-4(1-39) (SEQ ID No. 2).
[0092] In another embodiment of the invention the insulinotropic
agent is derived from a peptide having a length between 28 and 45
amino acid residues in which 22 out of the first 28 amino acid
residues are identical to those found in corresponding positions in
GLP-1(7-37) or in corresponding positions in Exendin-4(1-39).
[0093] In another embodiment of the invention the insulinotropic
agent is selected from a peptide comprising the amino acid sequence
of the formula (II):
Xaa.sub.7-Xaa.sub.8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Xaa.sub.16-Ser-Xaa-
.sub.18-Xaa.sub.19-Xaa.sub.20-Glu-Xaa.sub.22-Xaa.sub.23-Ala-Xaa.sub.25-Xaa-
.sub.26-Xaa.sub.27-Phe-Ile-Xaa.sub.30-Trp-Leu-Xaa.sub.33-Xaa.sub.34-Xaa.su-
b.35-Xaa.sub.36-Xaa.sub.37-Xaa.sub.38-Xaa.sub.39-Xaa.sub.40-Xaa.sub.41-Xaa-
.sub.42-Xaa.sub.43-Xaa.sub.44-Xaa.sub.45-Xaa.sub.46 Formula (II)
(SEQ ID No: 3) wherein Xaa.sub.7 is L-histidine, D-histidine,
desamino-histidine, 2-amino-histidine, .beta.-hydroxy-histidine,
homohistidine, N.sup..alpha.-acetyl-histidine,
.alpha.-fluoromethyl-histidine, .alpha.-methyl-histidine,
3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine; Xaa.sub.8
is Ala, D-Ala, Gly, Val, Leu, Ile, Lys, Aib,
(1-aminocyclopropyl)carboxylic acid, (1-aminocyclobutyl)carboxylic
acid, (1-aminocyclopentyl)carboxylic acid,
(1-aminocyclohexyl)carboxylic acid, (1-aminocycloheptyl)carboxylic
acid, or (1-aminocyclooctyl)carboxylic acid; Xaa.sub.16 is Val or
Leu; Xaa.sub.18 is Ser, Lys or Arg; Xaa.sub.19 is Tyr or Gln;
Xaa.sub.20 is Leu or Met; Xaa.sub.22 is Gly, Glu or Aib; Xaa.sub.23
is Gln, Glu, Lys or Arg; Xaa.sub.25 is Ala or Val; Xaa.sub.26 is
Lys, Glu or Arg; Xaa.sub.27 is Glu or Leu; Xaa.sub.30 is Ala, Glu
or Arg; Xaa.sub.33 is Val or Lys; Xaa.sub.34 is Lys, Glu, Asn or
Arg; Xaa.sub.35 is Gly or Aib; Xaa.sub.36 is Arg, Gly or Lys;
Xaa.sub.37 is Gly, Ala, Glu, Pro, Lys, amide or is absent;
Xaa.sub.38 is Lys, Ser, amide or is absent. Xaa.sub.39 is Ser, Lys,
amide or is absent; Xaa.sub.40 is Gly, amide or is absent;
Xaa.sub.41 is Ala, amide or is absent; Xaa.sub.42 is Pro, amide or
is absent; Xaa.sub.43 is Pro, amide or is absent; Xaa.sub.44 is
Pro, amide or is absent; Xaa.sub.45 is Ser, amide or is absent;
Xaa.sub.46 is amide or is absent; provided that if Xaa.sub.38,
Xaa.sub.39, Xaa.sub.40, Xaa.sub.41, Xaa.sub.42, Xaa.sub.43,
Xaa.sub.44, Xaa.sub.45 or Xaa.sub.46 is absent then each amino acid
residue downstream is also absent.
[0094] In another embodiment of the invention the insulinotropic
agent is a peptide comprising the amino acid sequence of formula
(III):
Xaa.sub.7-Xaa.sub.8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Xaa.sub.18-Tyr-Le-
u-Glu-Xaa.sub.22-Xaa.sub.23-Ala-Ala-Xaa.sub.26-Glu-Phe-Ile-Xaa.sub.30-Trp--
Leu-Val-Xaa.sub.34-Xaa.sub.35-Xaa.sub.36-Xaa.sub.37-Xaa.sub.38
Formula (III) (SEQ ID No: 4) wherein Xaa.sub.7 is L-histidine,
D-histidine, desamino-histidine, 2-amino-histidine,
.beta.-hydroxy-histidine, homohistidine,
N.sup..alpha.-acetyl-histidine, .alpha.-fluoromethyl-histidine,
.alpha.-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or
4-pyridylalanine; Xaa.sub.8 is Ala, D-Ala, Gly, Val, Leu, Ile, Lys,
Aib, (1-aminocyclopropyl)carboxylic acid,
(1-aminocyclobutyl)carboxylic acid, (1-aminocyclopentyl)carboxylic
acid, (1-aminocyclohexyl)carboxylic acid,
(1-aminocycloheptyl)carboxylic acid, or
(1-aminocyclooctyl)carboxylic acid; Xaa.sub.18 is Ser, Lys or Arg;
Xaa.sub.22 is Gly, Glu or Aib; Xaa.sub.23 is Gln, Glu, Lys or Arg;
Xaa.sub.26 is Lys, Glu or Arg; Xaa.sub.30 is Ala, Glu or Arg;
Xaa.sub.34 is Lys, Glu or Arg; Xaa.sub.35 is Gly or Aib; Xaa.sub.36
is Arg or Lys; Xaa.sub.37 is Gly, Ala, Glu or Lys; Xaa.sub.38 is
Lys, NH.sub.2 or is absent.
[0095] In another embodiment of the invention the insulinotropic
agent is selected from GLP-1(7-35), GLP-1(7-36), GLP-1(7-36)-amide,
GLP-1(7-37), GLP-1(7-38), GLP-1(7-39), GLP-1(7-40), GLP-1(7-41) or
an analogue thereof.
[0096] In another embodiment of the invention the insulinotropic
agent comprises no more than fifteen amino acid residues which have
been exchanged, added or deleted as compared to GLP-1(7-37) (SEQ ID
No. 1), or no more than ten amino acid residues which have been
exchanged, added or deleted as compared to GLP-1(7-37) (SEQ ID No.
1).
[0097] In another embodiment of the invention the insulinotropic
agent comprises no more than six amino acid residues which have
been exchanged, added or deleted as compared to GLP-1(7-37) (SEQ ID
No. 1).
[0098] In another embodiment of the invention the insulinotropic
agent comprises no more than 4 amino acid residues which are not
encoded by the genetic code.
[0099] In another embodiment of the invention the insulinotropic
agent comprises an Aib residue as the second amino acid residue
from the N-terminal.
[0100] In another embodiment of the invention the N-terminal amino
acid residue (position 7 in formulae II and III) of said
insulinotropic agent is selected from the group consisting of
D-histidine, desamino-histidine, 2-amino-histidine,
.beta.-hydroxy-histidine, homohistidine,
N.sup..alpha.-acetyl-histidine, .alpha.-fluoromethyl-histidine,
.alpha.-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine and
4-pyridylalanine.
[0101] In another embodiment of the invention the insulinotropic
agent is selected from the group consisting of
[Arg.sup.34]GLP-1(7-37), [Arg.sup.26,34]GLP-1(7-37)Lys,
[Lys.sup.36Arg.sup.26,34]GLP-1(7-36), [Aib.sup.8,22,35]GLP-1(7-37),
[Aib.sup.8,35]GLP-1(7-37), [Aib.sup.8,22]GLP-1(7-37),
[Aib.sup.8,22,35 Arg.sup.26,34]GLP-1(7-37)Lys,
[Aib.sup.8,35Arg.sup.26,34]GLP-1(7-37)Lys,
[Aib.sup.8,22Arg.sup.26,34]GLP-1(7-37)Lys,
[Aib.sup.8,22,35Arg.sup.26,34]GLP-1(7-37)Lys,
[Aib.sup.8,35Arg.sup.26,34]GLP-1(7-37)Lys,
[Aib.sup.8,22,35Arg.sup.26]GLP-1(7-37)Lys,
[Aib.sup.8,35Arg.sup.26]GLP-1(7-37)Lys,
[Aib.sup.8,22Arg.sup.26]GLP-1(7-37)Lys,
[Aib.sup.8,22,35Arg.sup.34]GLP-1(7-37)Lys,
[Aib.sup.8,35Arg.sup.34]GLP-1(7-37)Lys,
[Aib.sup.8,22Arg.sup.34]GLP-1(7-37)Lys,
[Aib.sup.8,22,35Ala.sup.37]GLP-1(7-37)Lys,
[Aib.sup.8,35Ala.sup.37]GLP-1(7-37)Lys,
[Aib.sup.8,22Ala.sup.37]GLP-1(7-37)Lys,
[Aib.sup.8,22,35Lys.sup.37]GLP-1(7-37),
[Aib.sup.8,35Lys.sup.37]GLP-1(7-37),
[Aib.sup.8,22Lys.sup.37]GLP-1(7-37) or derivatives thereof which
has been amidated on the C-terminal.
[0102] In another embodiment of the invention the insulinotropic
agent comprises at least one Aib residue.
[0103] In another embodiment of the invention the insulinotropic
agent contains two Aib residues.
[0104] In another embodiment of the invention the insulinotropic
agent comprises a serine residue at position 18 relative to
GLP-1(7-37) (SEQ ID: No. 1), corresponding to position 12 relative
to Exendin-4(1-39).
[0105] In another embodiment of the invention the insulinotropic
agent comprises a tyrosine residue at position 19 relative to
GLP-1(7-37) (SEQ ID. No. 1), corresponding to position 13 relative
to Exendin-4(1-39).
[0106] In another embodiment of the invention the insulinotropic
agent comprises a glycine residue at position 22 relative to
GLP-1(7-37) (SEQ ID. No. 1), corresponding to position 16 relative
to Exendin-4(1-39).
[0107] In another embodiment of the invention the insulinotropic
agent comprises a glutamine residue at position 23 relative to
GLP-1(7-37) (SEQ ID. No. 1), corresponding to position 17 relative
to Exendin-4(1-39).
[0108] In another embodiment of the invention the insulinotropic
agent comprises a lysine residue at position 26 relative to
GLP-1(7-37) (SEQ ID. No. 1), corresponding to position 20 relative
to Exendin-4(1-39).
[0109] In another embodiment of the invention the insulinotropic
agent comprises a glutamate residue at position 27 relative to
GLP-1(7-37) (SEQ ID. No. 1), corresponding to position 21 relative
to Exendin-4(1-39).
[0110] In another embodiment of the invention the insulinotropic
agent is exendin-4(1-39).
[0111] In another embodiment of the invention the insulinotropic
agent is ZP-10, i.e.
[Ser.sup.38Lys.sup.39]Exendin-4(1-39)LysLysLysLysLys-amide (SEQ ID
No. 5).
[0112] In another embodiment of the invention the insulinotropic
agent is attached to Y--C*-Q or Q via the amino acid residue in
position 25 to 45 relative to the amino acid sequence SEQ ID. No
1.
[0113] In another embodiment of the invention the insulinotropic
agent is attached to Y--C*-Q or Q via an amino acid residue
selected from one of the 10 C-terminal amino acid residues.
[0114] In another embodiment of the invention the insulinotropic
agent is attached to Y--C*-Q or Q via the amino acid residue in
position 23, 26, 34, 36 or 38 relative to the amino acid sequence
SEQ ID No:1.
[0115] In another embodiment of the invention the insulinotropic
agent is attached to Y--C*-Q or Q via the amino acid residue in
position 17, 20, 28, 30 or 32 relative to the amino acid sequence
SEQ ID No:2.
[0116] In another embodiment of the invention the insulinotropic
agent is attached to Y--C*-Q or Q via the C-terminal amino acid
residue.
[0117] In another embodiment of the invention the insulinotropic
agent is attached to Y--C*-Q or Q via a carboxyl group, an amino
group, a keto group, a hydroxyl group, a thiol group or a hydrazide
group.
[0118] In another embodiment of the invention the insulinotropic
agent is attached to Y--C*-Q or Q via a the epsilon-amino group on
a lysine residue.
[0119] In another embodiment of the invention the insulinotropic
agent comprises only one lysine residue.
[0120] In another embodiment of the invention the insulinotropic
agent comprises only one lysine residue which is the C-terminal
amino acid residue of said insulinotropic agent.
[0121] In another embodiment the compound according to the present
invention has an EC.sub.50 of less than 1000 pM, less than 500 pM,
less than 300 pM, less than 200 pM, less than 100 pM, less than 50
pM or less than 10 pM as determined by the functional receptor
assay disclosed herein.
[0122] In another embodiment the compound according to the present
invention is selected from the group consisting of [0123]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has
a molecular weight of approximately 20 kDa, [0124]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 20 kDa, [0125]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 5 kDa, [0126]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 2 kDa, [0127]
N.sup..epsilon.26-(3-(mPEGyl)propionyl)[Aib.sup.8,Glu.sup.22,30,Lys.sup.3-
3,Asn.sup.34,Gly.sup.35,36,Pro.sup.37]GLP-1(7-37)ylSerSerGly
AlaProProProSer amide wherein mPEGyl is polydisperse and has a
molecular weight of approximately 2 kDa, [0128]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl)Lysinamide) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 750 Da, [0129]
N.sup..epsilon.-[Aib.sup.8,22,35]GLP-1(7-37)yl(S.sup..epsilon.-(1-mPEGylp-
ropyl-2,5-dioxo-pyrrolidin-3-yl)Cysteinamide wherein mPEGyl is
polydisperse and has a molecular weight of approximately 5000 Da,
[0130] N.sup..alpha.-(3-(3H-imidazol-4-yl)-propionyl
[Aib.sup.22,35,Arg.sup.26,34]GLP-1-(8-37))yl(N.sup..epsilon.-(3-(mPEGyl)p-
ropionyl)Lysinamide) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 2000 Da, [0131]
N.sup..epsilon.26-(3-(mPEGyl)propionyl)[Arg.sup.34]GLP-1-(7-37)
wherein mPGyl is polydisperse and has a molecular weight of
approximately 2 kDa, and [0132]
(S)--N--((S)-5-(N--((S)-5-carbamoyl-5-(mPEGylpropionylamino)pentyl)carbam-
oyl)-5-(mPEGylpropionylamino)pentyl)-5-(N.sup..alpha.7-(3-(4-imidazolyl)pr-
opionyl)[Aib.sup.22,35,Arg.sup.26,34]GLP-1-(8-37)yl)-2-(mPEGylpropionylami-
no)hexanoic amide wherein mPEGyl is polydisperse and has a
molecular weight of approximately 750 Da,
[0133] In another embodiment the compound according to the present
invention is selected from the group consisting of [0134]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has
a molecular weight of approximately 2 kDa, [0135]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has
a molecular weight of approximately 5 kDa, [0136]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has
a molecular weight of approximately 20 kDa, [0137]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has
a molecular weight of approximately 40 kDa, [0138]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 2 kDa, [0139]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 5 kDa, [0140]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 20 kDa, [0141]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 40 kDa, [0142]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) Lys wherein mPEGyl is polydisperse and
has a molecular weight of approximately 2 kDa, [0143]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37)Lys wherein mPEGyl is polydisperse and
has a molecular weight of approximately 5 kDa, [0144]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37)Lys wherein mPEGyl is polydisperse and
has a molecular weight of approximately 20 kDa, [0145]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37)Lys wherein mPEGyl is polydisperse and
has a molecular weight of approximately 40 kDa, [0146]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37)Lys amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 2 kDa, [0147]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37)Lys amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 5 kDa, [0148]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) Lys amide wherein mPEGyl is
polydisperse and has a molecular weight of approximately 20 kDa,
[0149]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,35,Lys.sup.37]GLP-1(7-37) Lys amide wherein mPEGyl is
polydisperse and has a molecular weight of approximately 40 kDa,
[0150]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 2 kDa, [0151]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 5 kDa, [0152]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 20 kDa, [0153]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 40 kDa, [0154]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse and
has a molecular weight of approximately 2 kDa, [0155]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse and
has a molecular weight of approximately 5 kDa, [0156]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse and
has a molecular weight of approximately 20 kDa, [0157]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse and
has a molecular weight of approximately 40 kDa, [0158]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) Lys wherein mPEGyl is polydisperse and has
a molecular weight of approximately 2 kDa, [0159]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37)Lys wherein mPEGyl is polydisperse and has
a molecular weight of approximately 5 kDa, [0160]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37)Lys wherein mPEGyl is polydisperse and has
a molecular weight of approximately 20 kDa, [0161]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37)Lys wherein mPEGyl is polydisperse and has
a molecular weight of approximately 40 kDa, [0162]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37)Lys amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 2 kDa, [0163]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37)Lys amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 5 kDa, [0164]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) Lys amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 20 kDa, [0165]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,2-
2,Lys.sup.37]GLP-1(7-37) Lys amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 40 kDa, [0166]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 2 kDa, [0167]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 5 kDa, [0168]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 20 kDa, [0169]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is polydisperse and has a
molecular weight of approximately 40 kDa, [0170]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse and
has a molecular weight of approximately 2 kDa, [0171]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse and
has a molecular weight of approximately 5 kDa, [0172]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse and
has a molecular weight of approximately 20 kDa, [0173]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37) amide wherein mPEGyl is polydisperse and
has a molecular weight of approximately 40 kDa, [0174]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37)Lys wherein mPEGyl is polydisperse and has
a molecular weight of approximately 2 kDa, [0175]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37) Lys wherein mPEGyl is polydisperse and has
a molecular weight of approximately 5 kDa, [0176]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37) Lys wherein mPEGyl is polydisperse and has
a molecular weight of approximately 20 kDa, [0177]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37)Lys wherein mPEGyl is polydisperse and has
a molecular weight of approximately 40 kDa, [0178]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37)Lys amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 2 kDa, [0179]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37)Lys amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 5 kDa, [0180]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37)Lys amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 20 kDa, [0181]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,3-
5,Lys.sup.37]GLP-1(7-37)Lys amide wherein mPEGyl is polydisperse
and has a molecular weight of approximately 40 kDa, [0182]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 2 kDa, [0183]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 5 kDa, [0184]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 10 kDa, [0185]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 20 kDa, [0186]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 40 kDa, [0187]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 2 kDa, [0188]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 5 kDa, [0189]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 10 kDa, [0190]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 20 kDa, [0191]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 40 kDa, [0192]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37)Lys wherein mPEGyl is polydisperse and has a molecular weight
of approximately 2 kDa, [0193]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37)Lys wherein mPEGyl is polydisperse and has a molecular weight
of approximately 5 kDa, [0194]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37)Lys wherein mPEGyl is polydisperse and has a molecular weight
of approximately 10 kDa, [0195]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37)Lys wherein mPEGyl is polydisperse and has a molecular weight
of approximately 20 kDa, [0196]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37)Lys wherein mPEGyl is polydisperse and has a molecular weight
of approximately 40 kDa, [0197]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 2 kDa, [0198]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 5 kDa, [0199] N.sup..epsilon.37
(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(7-37)Lys
amide wherein mPEGyl is polydisperse and has a molecular weight of
approximately 10 kDa, [0200]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 20 kDa, [0201]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 40 kDa, [0202]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 2 kDa, [0203]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 5 kDa, [0204]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 10 kDa, [0205]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 20 kDa, [0206]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 40 kDa, [0207]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7) amide wherein mPEGyl is polydisperse and has a molecular weight
of approximately 2 kDa, [0208]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7) amide wherein mPEGyl is polydisperse and has a molecular weight
of approximately 5 kDa, [0209]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7) amide wherein mPEGyl is polydisperse and has a molecular weight
of approximately 10 kDa, [0210]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7) amide wherein mPEGyl is polydisperse and has a molecular weight
of approximately 20 kDa, [0211]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7) amide wherein mPEGyl is polydisperse and has a molecular weight
of approximately 40 kDa, [0212]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7)Lys wherein mPEGyl is polydisperse and has a molecular weight of
approximately 2 kDa, [0213]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7)Lys wherein mPEGyl is polydisperse and has a molecular weight of
approximately 5 kDa,
[0214]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]G-
LP-1(7-37)Lys wherein mPEGyl is polydisperse and has a molecular
weight of approximately 10 kDa, [0215]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7)Lys wherein mPEGyl is polydisperse and has a molecular weight of
approximately 20 kDa, [0216]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7)Lys wherein mPEGyl is polydisperse and has a molecular weight of
approximately 40 kDa, [0217]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 2 kDa, [0218]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 5 kDa, [0219]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 10 kDa, [0220]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 20 kDa, [0221]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,Lys.sup.37]GLP-1(7-3-
7)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 40 kDa, [0222]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 2 kDa, [0223]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 5 kDa, [0224]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 10 kDa, [0225]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 20 kDa, [0226]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7) wherein mPEGyl is polydisperse and has a molecular weight of
approximately 40 kDa, [0227]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7) amide wherein mPEGyl is polydisperse and has a molecular weight
of approximately 2 kDa, [0228]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7) amide wherein mPEGyl is polydisperse and has a molecular weight
of approximately 5 kDa, [0229]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7) amide wherein mPEGyl is polydisperse and has a molecular weight
of approximately 10 kDa, [0230]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7) amide wherein mPEGyl is polydisperse and has a molecular weight
of approximately 20 kDa, [0231]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7) amide wherein mPEGyl is polydisperse and has a molecular weight
of approximately 40 kDa, [0232]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7)Lys wherein mPEGyl is polydisperse and has a molecular weight of
approximately 2 kDa, [0233]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1-(7--
37)Lys wherein mPEGyl is polydisperse and has a molecular weight of
approximately 5 kDa, [0234]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7)Lys wherein mPEGyl is polydisperse and has a molecular weight of
approximately 10 kDa, [0235]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7)Lys wherein mPEGyl is polydisperse and has a molecular weight of
approximately 20 kDa, [0236]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7)Lys wherein mPEGyl is polydisperse and has a molecular weight of
approximately 40 kDa, [0237]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 2 kDa, [0238]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 5 kDa, [0239]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 10 kDa, [0240]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 20 kDa, and [0241]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,35,Lys.sup.37]GLP-1(7-3-
7)Lys amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 40 kDa.
[0242] In another embodiment the compound according to the present
invention is selected from the group consisting of [0243]
N.sup..epsilon.37-(3-(mPEGyl)propionyl) [Lys.sup.37]GLP-1-(7-37)
where mPEGyl is polydisperse and is polydisperse and has a Mw of
approx 750 Da, [0244] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is polydisperse and has
a Mw of approx 750 Da, [0245]
GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine where
mPEGyl is polydisperse and has a Mw of approx 750 Da, [0246]
GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysinamide
where mPEGyl is polydisperse and has a Mw of approx 750 Da, [0247]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 750 Da, [0248]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0249]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
750 Da, [0250]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)propionyl)) Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 750 Da, [0251] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 750 Da, [0252]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0253]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
750 Da, [0254]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)propionyl)) Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 750 Da, [0255] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse and
has a Mw of approx 750 Da, [0256]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0257]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propio-
nyl))Lysine where mPEGyl is polydisperse and has a Mw of approx 750
Da, [0258]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGy-
l)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 750 Da, [0259] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 750 Da, [0260]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0261]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
750 Da, [0262]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(m-
PEGyl)propionyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 750 Da, [0263] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 750 Da, [0264]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0265]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
750 Da, [0266]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl)) Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 750 Da, [0267] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 750 Da, [0268]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0269]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
750 Da, [0270]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(m-
PEGyl)propionyl)) Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 750 Da, [0271] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 750 Da, [0272]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0273]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
750 Da, [0274]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(m-
PEGyl)propionyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 750 Da, [0275] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0276]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0277]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl))Lysine where mPEGyl is polydisperse and has a Mw of
approx 750 Da, [0278]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 750 Da, [0279] N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 750 Da, [0280]
N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysina-
mide where mPEGyl is polydisperse and has a Mw of approx 750 Da
[0281] N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Aib.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 750 Da, [0282]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Aib.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0283]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0284]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0285]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
750 Da, [0286]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 750 Da, [0287] N.sup..epsilon.36-(3-(mPEGyl)propionyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 750 Da, [0288]
N.sup..epsilon.36-(3-(mPEGyl)propionyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0289]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 750 Da, [0290]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0291]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 750 Da, [0292]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysina-
mide where mPEGyl is polydisperse and has a Mw of approx 750 Da,
[0293] N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Ala.sup.8,
Arg.sup.26,34Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 750 Da, [0294]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Ala.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0295]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0296]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, [0297]
N.sup..epsilon.20-(3-(mPEGyl)propionyl)
[Lys.sup.20]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 750 Da, [0298]
N.sup..epsilon.32-(3-(mPEGyl)propionyl)
[Lys.sup.32]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 750 Da, [0299]
N.sup..epsilon.20-(3-(mPEGyl)propionyl)
[Lys.sup.20,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da, and [0300]
N.sup..epsilon.32-(3-(mPEGyl)propionyl)
[Lys.sup.32,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 750 Da.
[0301] In another embodiment the compound according to the present
invention is selected from the group consisting of [0302]
N.sup..epsilon.37-(3-(mPEGyl)propionyl) [Lys.sup.37]GLP-1-(7-37)
where mPEGyl is polydisperse and has a Mw of approx 2000 Da, [0303]
N.sup..epsilon.37-(3-(mPEGyl)propionyl) [Lys.sup.37]GLP-1-(7-37)
amide where mPEGyl is polydisperse and has a Mw of approx 2000 Da,
[0304] GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 2000 Da, [0305]
GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysinamide
where mPEGyl is polydisperse and has a Mw of approx 2000 Da, [0306]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 2000 Da, [0307]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0308]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
2000 Da, [0309]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 2000 Da, [0310] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 2000 Da, [0311]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0312]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
2000 Da, [0313]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 2000 Da, [0314] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse and
has a Mw of approx 2000 Da, [0315]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0316]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propio-
nyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
2000 Da, [0317]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGy-
l)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 2000 Da, [0318] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 2000 Da, [0319]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0320]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
2000 Da, [0321]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 2000 Da, [0322] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 2000 Da, [0323]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0324]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
2000 Da, [0325]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 2000 Da, [0326] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 2000 Da, [0327]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0328]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
2000 Da, [0329]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 2000 Da, [0330] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 2000 Da, [0331]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0332]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
2000 Da, [0333]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 2000 Da, [0334] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0335]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0336]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl))Lysine where mPEGyl is polydisperse and has a Mw of
approx 2000 Da, [0337]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 2000 Da, [0338] N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 2000 Da, [0339]
N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysina-
mide where mPEGyl is polydisperse and has a Mw of approx 2000 Da
[0340] N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Aib.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 2000 Da, [0341]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Aib.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0342]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0343]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0344]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
2000 Da, [0345]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 2000 Da, [0346] N.sup..epsilon.36-(3-(mPEGyl)propionyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 2000 Da, [0347]
N.sup..epsilon.36-(3-(mPEGyl)propionyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0348]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 2000 Da, [0349]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0350]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 2000 Da, [0351]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysina-
mide where mPEGyl is polydisperse and has a Mw of approx 2000 Da,
[0352] N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Ala.sup.8,
Arg.sup.26,34 Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 2000 Da, [0353]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Ala.sup.8,
Arg.sup.26,34Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0354]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0355]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, [0356]
N.sup..epsilon.20-(3-(mPEGyl)propionyl)
[Lys.sup.20]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 2000 Da, [0357]
N.sup..epsilon.32-(3-(mPEGyl)propionyl)
[Lys.sup.32]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 2000 Da, [0358]
N.sup..epsilon.20-(3-(mPEGyl)propionyl)
[Lys.sup.20,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da, and [0359]
N.sup..epsilon.32-(3-(mPEGyl)propionyl)
[Lys.sup.32,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 2000 Da.
[0360] In another embodiment the compound according to the present
invention is selected from the group consisting of [0361]
N.sup..epsilon.37-(3-(mPEGyl)propionyl) [Lys.sup.37]GLP-1-(7-37)
where mPEGyl is polydisperse and has a Mw of approx 5000 Da, [0362]
N.sup..epsilon.37-(3-(mPEGyl)propionyl) [Lys.sup.37]GLP-1-(7-37)
amide where mPEGyl is polydisperse and has a Mw of approx 5000 Da,
[0363] GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 5000 Da, [0364]
GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysinamide
where mPEGyl is polydisperse and has a Mw of approx 5000 Da, [0365]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 5000 Da, [0366]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0367]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
5000 Da, [0368]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 5000 Da, [0369] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 5000 Da, [0370]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0371]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
5000 Da, [0372]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 5000 Da, [0373] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse and
has a Mw of approx 5000 Da, [0374]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0375]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propio-
nyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
5000 Da, [0376]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGy-
l)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 5000 Da, [0377] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 5000 Da, [0378]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0379]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
5000 Da, [0380]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl)) Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 5000 Da, [0381] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 5000 Da, [0382]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0383]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
5000 Da, [0384]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 5000 Da, [0385] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 5000 Da, [0386]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0387]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
5000 Da, [0388]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 5000 Da, [0389] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 5000 Da, [0390]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0391]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
5000 Da, [0392]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mP-
EGyl)propionyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 5000 Da, [0393]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0394]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0395]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl))Lysine where mPEGyl is polydisperse and has a Mw of
approx 5000 Da, [0396]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 5000 Da, [0397] N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 5000 Da, [0398]
N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysina-
mide where mPEGyl is polydisperse and has a Mw of approx 5000 Da
[0399] N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Aib.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 5000 Da, [0400]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Aib.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0401]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0402]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0403]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
5000 Da, [0404]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 5000 Da, [0405] N.sup..epsilon.36-(3-(mPEGyl)propionyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 5000 Da, [0406]
N.sup..epsilon.36-(3-(mPEGyl)propionyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0407]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 5000 Da, [0408]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0409]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 5000 Da, [0410]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysina-
mide where mPEGyl is polydisperse and has a Mw of approx 5000 Da,
[0411] N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Ala.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 5000 Da, [0412]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Ala.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0413]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0414]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, [0415]
N.sup..epsilon.20-(3-(mPEGyl)propionyl)
[Lys.sup.20]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 5000 Da, [0416]
N.sup..epsilon.32-(3-(mPEGyl)propionyl)
[Lys.sup.32]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 5000 Da, [0417]
N.sup..epsilon.20-(3-(mPEGyl)propionyl)
[Lys.sup.20,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da, and [0418]
N.sup..epsilon.32-(3-(mPEGyl)propionyl)
[Lys.sup.32,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 5000 Da.
[0419] In another embodiment the compound according to the present
invention is selected from the group consisting of [0420]
N.sup..epsilon.37-(3-(mPEGyl)propionyl) [Lys.sup.37]GLP-1-(7-37)
where mPEGyl is polydisperse and has a Mw of approx 10 kDa, [0421]
N.sup..epsilon.37-(3-(mPEGyl)propionyl) [Lys.sup.37]GLP-1-(7-37)
amide where mPEGyl is polydisperse and has a Mw of approx 10 kDa,
[0422] GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 10 kDa, [0423]
GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysinamide
where mPEGyl is polydisperse and has a Mw of approx 10 kDa, [0424]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 10 kDa, [0425]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0426]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx 10
kDa, [0427]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)propionyl)) Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 10 kDa, [0428] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 10 kDa, [0429]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0430]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx 10
kDa, [0431]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 10 kDa, [0432] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse and
has a Mw of approx 10 kDa, [0433]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0434]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propio-
nyl))Lysine where mPEGyl is polydisperse and has a Mw of approx 10
kDa, [0435]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGy-
l)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 10 kDa, [0436] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 10 kDa, [0437]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0438]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
10 kDa, [0439]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(m-
PEGyl)propionyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 10 kDa, [0440] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 10 kDa, [0441]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0442]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
10 kDa, [0443]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 10 kDa, [0444] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 10 kDa, [0445]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0446]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
10 kDa, [0447]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(m-
PEGyl)propionyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 10 kDa, [0448] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 10 kDa, [0449]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0450]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
10 kDa, [0451]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(m-
PEGyl)propionyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 10 kDa, [0452] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0453]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0454]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl))Lysine where mPEGyl is polydisperse and has a Mw of
approx 10 kDa, [0455]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 10 kDa, [0456] N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 10 kDa, [0457]
N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysina-
mide where mPEGyl is polydisperse and has a Mw of approx 10 kDa
[0458] N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Aib.sup.8,
Arg.sup.26,34 Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 10 kDa, [0459]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Aib.sup.8,
Arg.sup.26,34Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0460]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0461]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0462]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
10 kDa, [0463]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 10 kDa, [0464] N.sup..epsilon.36-(3-(mPEGyl)propionyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 10 kDa, [0465]
N.sup..epsilon.36-(3-(mPEGyl)propionyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0466]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 10 kDa, [0467]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0468]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 10 kDa, [0469]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysina-
mide where mPEGyl is polydisperse and has a Mw of approx 10 kDa,
[0470] N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Ala.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 10 kDa, [0471]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Ala.sup.8, Arg.sup.26,34
Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is polydisperse and has
a Mw of approx 10 kDa, [0472]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0473]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, [0474]
N.sup..epsilon.20-(3-(mPEGyl)propionyl)
[Lys.sup.20]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 10 kDa, [0475]
N.sup..epsilon.32-(3-(mPEGyl)propionyl)
[Lys.sup.32]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 10 kDa, [0476]
N.sup..epsilon.20-(3-(mPEGyl)propionyl)
[Lys.sup.20,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa, and [0477]
N.sup..epsilon.32-(3-(mPEGyl)propionyl)
[Lys.sup.32,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 10 kDa.
[0478] In another embodiment the compound according to the present
invention is selected from the group consisting of [0479]
N.sup..epsilon.37-(3-(mPEGyl)propionyl) [Lys.sup.37]GLP-1-(7-37)
where mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0480]
N.sup..epsilon.37-(3-(mPEGyl)propionyl) [Lys.sup.37]GLP-1-(7-37)
amide where mPEGyl is polydisperse and has a Mw of approx 20 kDa,
[0481] GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0482]
GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysinamide
where mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0483]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0484]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0485]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx 20
kDa, [0486]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 20 kDa, [0487] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0488]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0489]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propi-
onyl))Lysine where mPEGyl is polydisperse and has a Mw of approx 20
kDa, [0490]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 20 kDa, [0491] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0492]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0493]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propio-
nyl))Lysine where mPEGyl is polydisperse and has a Mw of approx 20
kDa, [0494]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGy-
l)propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 20 kDa, [0495] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0496]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0497]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
20 kDa, [0498]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(m-
PEGyl)propionyl)) Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 20 kDa, [0499] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0500]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0501]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
20 kDa, [0502]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 20 kDa, [0503] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0504]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0505]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
20 kDa, [0506]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(m-
PEGyl)propionyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 20 kDa, [0507] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0508]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0509]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pro-
pionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
20 kDa, [0510]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(m-
PEGyl)propionyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 20 kDa, [0511] N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0512]
N.sup..epsilon.37-(3-(mPEGyl)propionyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0513]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl))Lysine where mPEGyl is polydisperse and has a Mw of
approx 20 kDa, [0514]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
propionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 20 kDa, [0515] N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0516]
N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysina-
mide where mPEGyl is polydisperse and has a Mw of approx 20 kDa
[0517] N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Aib.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0518]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Aib.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0519]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0520]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0521]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
20 kDa, [0522]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)pr-
opionyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 20 kDa, [0523] N.sup..epsilon.36-(3-(mPEGyl)propionyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0524]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Arg.sup.26,34
Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is polydisperse and has
a Mw of approx 20 kDa, [0525]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0526]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0527]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0528]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)propionyl))Lysina-
mide where mPEGyl is polydisperse and has a Mw of approx 20 kDa,
[0529] N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Ala.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0530]
N.sup..epsilon.36-(3-(mPEGyl)propionyl) [Ala.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0531]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0532]
N.sup..epsilon.34-(3-(mPEGyl)propionyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0533]
N.sup..epsilon.20-(3-(mPEGyl)propionyl)
[Lys.sup.20]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0534]
N.sup..epsilon.32-(3-(mPEGyl)propionyl)
[Lys.sup.32]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0535]
N.sup..epsilon.20-(3-(mPEGyl)propionyl)
[Lys.sup.20,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, and [0536]
N.sup..epsilon.32-(3-(mPEGyl)propionyl)
[Lys.sup.32,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa.
[0537] In another embodiment the compound according to the present
invention is selected from the group consisting of [0538]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl) [Lys.sup.37]GLP-1-(7-37)
where mPEGyl is polydisperse and has a Mw of approx 30 kDa, [0539]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl) [Lys.sup.37]GLP-1-(7-37)
amide where mPEGyl is polydisperse and has a Mw of approx 30 kDa,
[0540] GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)butanoyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 30 kDa, [0541]
GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)butanoyl))Lysinamide
where mPEGyl is polydisperse and has a Mw of approx 30 kDa, [0542]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 30 kDa, [0543]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0544]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)butan-
oyl))Lysine where mPEGyl is polydisperse and has a Mw of approx 30
kDa, [0545]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)butanoyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 30 kDa, [0546] N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 30 kDa, [0547]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0548]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)butan-
oyl))Lysine where mPEGyl is polydisperse and has a Mw of approx 30
kDa, [0549]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEG-
yl)butanoyl)) Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 30 kDa, [0550] N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse and
has a Mw of approx 30 kDa, [0551]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0552]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)butano-
yl))Lysine where mPEGyl is polydisperse and has a Mw of approx 30
kDa, [0553]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGy-
l)butanoyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 30 kDa, [0554] N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 30 kDa, [0555]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0556]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)but-
anoyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
30 kDa, [0557]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mP-
EGyl)butanoyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 30 kDa, [0558] N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 30 kDa, [0559]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0560]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)bu-
tanoyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
30 kDa, [0561]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(-
mPEGyl)butanoyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 30 kDa, [0562] N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 30 kDa, [0563]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0564]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)but-
anoyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
30 kDa, [0565]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mP-
EGyl)butanoyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 30 kDa, [0566] N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 30 kDa, [0567]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0568]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)but-
anoyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
30 kDa, [0569]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mP-
EGyl)butanoyl))Lysinamide where mPEGyl is polydisperse and has a Mw
of approx 30 kDa, [0570] N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0571]
N.sup..epsilon.37-(3-(mPEGyl)butanoyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0572]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
butanoyl))Lysine where mPEGyl is polydisperse and has a Mw of
approx 30 kDa, [0573]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)-
butanoyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 30 kDa, [0574] N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)butanoyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 30 kDa, [0575]
N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)butanoyl))Lysinam-
ide where mPEGyl is polydisperse and has a Mw of approx 30 kDa,
[0576] N.sup..epsilon.36-(3-(mPEGyl)butanoyl) [Aib.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 30 kDa, [0577]
N.sup..epsilon.36-(3-(mPEGyl)butanoyl) [Aib.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0578]
N.sup..epsilon.34-(3-(mPEGyl)butanoyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0579]
N.sup..epsilon.34-(3-(mPEGyl)butanoyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0580]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)bu-
tanoyl))Lysine where mPEGyl is polydisperse and has a Mw of approx
30 kDa, [0581]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(-
mPEGyl)butanoyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 30 kDa, [0582] N.sup..epsilon.36-(3-(mPEGyl)butanoyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 30 kDa, [0583]
N.sup..epsilon.36-(3-(mPEGyl)butanoyl)
[Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0584]
N.sup..epsilon.34-(3-(mPEGyl)butanoyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 30 kDa, [0585]
N.sup..epsilon.34-(3-(mPEGyl)butanoyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0586]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)butanoyl))Lysine
where mPEGyl is polydisperse and has a Mw of approx 30 kDa, [0587]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(3-(mPEGyl)butanoyl))Lysinam-
ide where mPEGyl is polydisperse and has a Mw of approx 30 kDa,
[0588] N.sup..epsilon.36-(3-(mPEGyl)butanoyl) [Ala.sup.8,
Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 30 kDa, [0589]
N.sup..epsilon.36-(3-(mPEGyl)butanoyl) [Ala.sup.8,
Arg.sup.26,34Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0590]
N.sup..epsilon.34-(3-(mPEGyl)butanoyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0591]
N.sup..epsilon.34-(3-(mPEGyl)butanoyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, [0592]
N.sup..epsilon.20-(3-(mPEGyl)butanoyl) [Lys.sup.20]Exendin-4-(1-39)
amide where mPEGyl is polydisperse and has a Mw of approx 30 kDa,
[0593] N.sup..epsilon.32-(3-(mPEGyl)butanoyl)
[Lys.sup.32]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 30 kDa, [0594]
N.sup..epsilon.20-(3-(mPEGyl)butanoyl)
[Lys.sup.20,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa, and [0595]
N.sup..epsilon.32-(3-(mPEGyl)butanoyl)
[Lys.sup.32,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 30 kDa.
[0596] In another embodiment the compound according to the present
invention is selected from the group consisting of [0597]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse and has a Mw
of approx 20 kDa, [0598]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is polydisperse and has
a Mw of approx 20 kDa, [0599]
GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl-
))Lysine where mPEGyl is polydisperse and has a Mw of approx 20
kDa, [0600]
GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mPEGylcarbonylamino)-
hexanoyl))Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 20 kDa, [0601]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0602]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0603]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mP-
EGylcarbonylamino)hexanoyl))Lysine where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0604]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mP-
EGylcarbonylamino)hexanoyl))Lysinamide where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0605]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0606]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0607]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mP-
EGylcarbonylamino)hexanoyl))Lysine where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0608]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mP-
EGylcarbonylamino)hexanoyl))Lysinamide where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0609]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0610]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0611]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mPE-
Gylcarbonylamino)hexanoyl))Lysine where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0612]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mPE-
Gylcarbonylamino)hexanoyl))Lysinamide where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0613]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0614]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0615]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(-
mPEGylcarbonylamino)hexanoyl)) Lysine where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0616]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(-
mPEGylcarbonylamino)hexanoyl)) Lysinamide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0617]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0618]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0619]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di--
(mPEGylcarbonylamino)hexanoyl))Lysine where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0620]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di--
(mPEGylcarbonylamino)hexanoyl))Lysinamide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0621]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0622]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0623]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(-
mPEGylcarbonylamino)hexanoyl)) Lysine where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0624]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(-
mPEGylcarbonylamino)hexanoyl))Lysinamide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0625]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0626]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0627]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(-
mPEGylcarbonylamino)hexanoyl))Lysine where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0628]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(-
mPEGylcarbonylamino)hexanoyl))Lysinamide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0629]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0630]
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0631]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-d-
i-(mPEGylcarbonylamino)hexanoyl))Lysine where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0632]
N.sup..alpha.-[Aib.sup.8,22,35s]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6--
di-(mPEGylcarbonylamino)hexanoyl))Lysinamide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0633]
N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mPEGylcarbonyl-
amino)hexanoyl))Lysine where mPEGyl is polydisperse and has a Mw of
approx 20 kDa, [0634] N.sup..alpha.-[Aib.sup.8, Arg.sup.26
34]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mPEGylcarbonylamino)hexan-
oyl))Lysinamide where mPEGyl is polydisperse and has a Mw of approx
20 kDa, [0635]
N.sup..epsilon.36-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8, Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0636]
N.sup..epsilon.36-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8, Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0637]
N.sup..epsilon.34-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0638]
N.sup..epsilon.34-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0639]
N.sup..alpha.-[Arg.sup.26
34]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mPEGylcarbonylamino)hexan-
oyl))Lysine where mPEGyl is polydisperse and has a Mw of approx 20
kDa, [0640] N.sup..alpha.-[Arg.sup.26
34]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mPEGylcarbonylamino)hexan-
oyl)) Lysinamide where mPEGyl is polydisperse and has a Mw of
approx 20 kDa, [0641]
N.sup..epsilon.36-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Arg.sup.26,34Lys.sup.36]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0642]
N.sup..epsilon.36-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Arg.sup.26,34 Lys.sup.36]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0643]
N.sup..epsilon.34-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0644]
N.sup..epsilon.34-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0645]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mPEGylcarbonyl-
amino)hexanoyl)) Lysine where mPEGyl is polydisperse and has a Mw
of approx 20 kDa, [0646] N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-((2S)-2,6-di-(mPEGylcarbonyl-
amino)hexanoyl))Lysinamide where mPEGyl is polydisperse and has a
Mw of approx 20 kDa, [0647]
N.sup..epsilon.36-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Ala.sup.8, Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0648]
N.sup..epsilon.36-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Ala.sup.8, Arg.sup.26,34Lys.sup.36]GLP-1-(7-37) amide where mPEGyl
is polydisperse and has a Mw of approx 20 kDa, [0649]
N.sup..epsilon.34-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0650]
N.sup..epsilon.34-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0651]
N.sup..epsilon.20-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Lys.sup.20]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0652]
N.sup..epsilon.32-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Lys.sup.32]Exendin-4-(1-39) amide where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0653]
N.sup..epsilon.20-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Lys.sup.20,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, and [0654]
N.sup..epsilon.32-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)
[Lys.sup.32,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa.
[0655] In another embodiment the compound according to the present
invention is selected from the group consisting of [0656]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Lys.sup.37]GLP-1-(7-37) where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0657]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is polydisperse
and has a Mw of approx 20 kDa, [0658]
GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)pr-
op-2-yloxy)butanoyl))Lysine where mPEGyl is polydisperse and has a
Mw of approx 20 kDa, [0659]
GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)pr-
op-2-yloxy)butanoyl))Lysinamide where mPEGyl is polydisperse and
has a Mw of approx 20 kDa, [0660]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0661]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0662]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEG-
ylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysine where mPEGyl
is polydisperse and has a Mw of approx 20 kDa, [0663]
N.sup..alpha.-[Aib.sup.35]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEG-
ylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysinamide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0664]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0665]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0666]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEG-
ylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysine where mPEGyl
is polydisperse and has a Mw of approx 20 kDa, [0667]
N.sup..alpha.-[Aib.sup.22]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEG-
ylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysinamide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0668]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0669]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0670]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEGy-
lethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysine where mPEGyl
is polydisperse and has a Mw of approx 20 kDa, [0671]
N.sup..alpha.-[Aib.sup.8]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEGy-
lethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysinamide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0672]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0673]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0674]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mP-
EGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysine where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0675]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mP-
EGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysinamide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0676]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0677]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl
is polydisperse and has a Mw of approx 20 kDa, [0678]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(m-
PEGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysine where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0679]
N.sup..alpha.-[Aib.sup.22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(m-
PEGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysinamide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0680]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0681]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0682]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mP-
EGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysine where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0683]
N.sup..alpha.-[Aib.sup.8,35]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mP-
EGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysinamide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0684]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0685]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,22,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0686]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mP-
EGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysine where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0687]
N.sup..alpha.-[Aib.sup.8,22]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mP-
EGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysinamide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0688]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0689]
N.sup..epsilon.37-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1-(7-37) amide where mPEGyl
is polydisperse and has a Mw of approx 20 kDa, [0690]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis-
(mPEGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysine where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0691]
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis-
(mPEGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysinamide
where mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0692]
N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEGylethylamino-
carbonyloxy)prop-2-yloxy)butanoyl))Lysine where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0693]
N.sup..alpha.-[Aib.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEGylethylamino-
carbonyloxy)prop-2-yloxy)butanoyl))Lysinamide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0694]
N.sup..epsilon.36
(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl)
[Aib.sup.8, Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0695]
N.sup..epsilon.36
(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl)
[Aib.sup.8, Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0696]
N.sup..epsilon.34-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl
is polydisperse and has a Mw of approx 20 kDa, [0697]
N.sup..epsilon.34-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Aib.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0698]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(m-
PEGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysine where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0699]
N.sup..alpha.-[Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(m-
PEGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl))Lysinamide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0700]
N.sup..epsilon.36-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0701]
N.sup..epsilon.36-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Arg.sup.26,34 Lys.sup.36]GLP-1-(7-37) amide where mPEGyl
is polydisperse and has a Mw of approx 20 kDa, [0702]
N.sup..epsilon.34-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0703]
N.sup..epsilon.34-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0704]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEGylethylamino-
carbonyloxy)prop-2-yloxy)butanoyl))Lysine where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0705]
N.sup..alpha.-[Ala.sup.8,
Arg.sup.26,34]GLP-1-(7-37)yl(N.sup..epsilon.-(4-(1,3-bis(mPEGylethylamino-
carbonyloxy)prop-2-yloxy)butanoyl))Lysinamide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0706]
N.sup..epsilon.36-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Ala.sup.8, Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0707]
N.sup..epsilon.36-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Ala.sup.8, Arg.sup.26,34,Lys.sup.36]GLP-1-(7-37) amide
where mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0708]
N.sup..epsilon.34-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) where mPEGyl
is polydisperse and has a Mw of approx 20 kDa, [0709]
N.sup..epsilon.34-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Ala.sup.8,Lys.sup.26,Lys.sup.34]GLP-1-(7-37) amide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0710]
N.sup..epsilon.20-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Lys.sup.20]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0711]
N.sup..epsilon.32-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Lys.sup.32]Exendin-4-(1-39)amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa, [0712]
N.sup..epsilon.20-(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)bu-
tanoyl) [Lys.sup.20,Arg.sup.12,27]Exendin-4-(1-39) amide where
mPEGyl is polydisperse and has a Mw of approx 20 kDa, [0713]
N.sup..epsilon.32
(4-(1,3-bis(mPEGylethylaminocarbonyloxy)prop-2-yloxy)butanoyl)
[Lys.sup.32,Arg.sup.12,27]Exendin-4-(1-39) amide where mPEGyl is
polydisperse and has a Mw of approx 20 kDa,
[0714] The compounds of the present invention can be produced by
classical peptide synthesis, e.g. solid phase peptide synthesis
using t-Boc or Fmoc chemistry or other well established techniques,
see e.g. Green and Wuts, "Protecting Groups in Organic Synthesis",
John Wiley & Sons, 1999. These methods are preferred when the
insulinotropic agent is a peptide comprising non-natural amino acid
residues.
[0715] When the insulinotropic agent is a polypeptide comprising
only amino acid residues encoded by the genetic code, the
polypeptides can also be produced by a method which comprises
culturing a host cell containing a DNA sequence encoding the
polypeptide and capable of expressing the polypeptide in a suitable
nutrient medium under conditions permitting the expression of the
peptide, after which the resulting peptide is recovered from the
culture and then derivatized to the compound of formula (I).
[0716] The medium used to culture the cells may be any conventional
medium suitable for growing the host cells, such as minimal or
complex media containing appropriate supplements. Suitable media
are available from commercial suppliers or may be prepared
according to published recipes (e.g. in catalogues of the American
Type Culture Collection). The peptide produced by the cells may
then be recovered from the culture medium by conventional
procedures including separating the host cells from the medium by
centrifugation or filtration. For extracellular products the
proteinaceous components of the supernatant are isolated by
filtration, column chromatography or precipitation, e.g.
microfiltation, ultrafiltration, isoelectric precipitation,
purification by a variety of chromatographic procedures, e.g. ion
exchange chromatography, hydrophobic interaction chromatography,
gel filtration chromatography, affinity chromatography, or the
like, dependent on the type of polypeptide in question. For
intracellular or periplasmic products the cells isolated from the
culture medium are disintegrated or permeabilised and extracted to
recover the product polypeptide or precursor thereof.
[0717] The DNA sequence encoding the therapeutic polypeptide may
suitably be of genomic or cDNA origin, for instance obtained by
preparing a genomic or cDNA library and screening for DNA sequences
coding for all or part of the peptide by hybridisation using
synthetic oligonucleotide probes in accordance with standard
techniques (see, for example, Sambrook, J, Fritsch, E F and
Maniatis, T, Molecular Cloning: A Laboratory Manual, Cold Spring
Harbor Laboratory Press, New York, 1989). The DNA sequence encoding
the polypeptide may also be prepared synthetically by established
standard methods, e.g. the phosphoamidite method described by
Beaucage and Caruthers, Tetrahedron Letters 22 (1981), 1859-1869,
or the method described by Matthes et al., EMBO Journal 3 (1984),
801-805. The DNA sequence may also be prepared by polymerase chain
reaction using specific primers, for instance as described in U.S.
Pat. No. 4,683,202 or Saiki et al., Science 239 (1988),
487-491.
[0718] The DNA sequence may be inserted into any vector which may
conveniently be subjected to recombinant DNA procedures, and the
choice of vector will often depend on the host cell into which it
is to be introduced. Thus, the vector may be an autonomously
replicating vector, i.e. a vector which exists as an
extrachromosomal entity, the replication of which is independent of
chromosomal replication, e.g. a plasmid. Alternatively, the vector
may be one which, when introduced into a host cell, is integrated
into the host cell genome and replicated together with the
chromosome(s) into which it has been integrated.
[0719] The vector is preferably an expression vector in which the
DNA sequence encoding the polypeptide is operably linked to
additional segments required for transcription of the DNA, such as
a promoter. The promoter may be any DNA sequence which shows
transcriptional activity in the host cell of choice and may be
derived from genes encoding proteins either homologous or
heterologous to the host cell. Examples of suitable promoters for
directing the transcription of the DNA encoding the peptide of the
invention in a variety of host cells are well known in the art, cf.
for instance Sambrook et al., supra.
[0720] The DNA sequence encoding the polypeptide may also, if
necessary, be operably connected to a suitable terminator,
polyadenylation signals, transcriptional enhancer sequences, and
translational enhancer sequences. The recombinant vector of the
invention may further comprise a DNA sequence enabling the vector
to replicate in the host cell in question.
[0721] The vector may also comprise a selectable marker, e.g. a
gene the product of which complements a defect in the host cell or
one which confers resistance to a drug, e.g. ampicillin, kanamycin,
tetracyclin, chloramphenicol, neomycin, hygromycin or methotrexate.
For large scale manufacture the selectable marker preferably is not
antibiotic resistance, e.g. antibiotic resistance genes in the
vector are preferably excised when the vector is used for large
scale manufacture. Methods for eliminating antibiotic resistance
genes from vectors are known in the art, see e.g. U.S. Pat. No.
6,358,705 which is incorporated herein by reference.
[0722] To direct a parent peptide of the present invention into the
secretory pathway of the host cells, a secretory signal sequence
(also known as a leader sequence, prepro sequence or pre sequence)
may be provided in the recombinant vector. The secretory signal
sequence is joined to the DNA sequence encoding the peptide in the
correct reading frame. Secretory signal sequences are commonly
positioned 5' to the DNA sequence encoding the peptide. The
secretory signal sequence may be that normally associated with the
peptide or may be from a gene encoding another secreted
protein.
[0723] The procedures used to ligate the DNA sequences coding for
the present peptide, the promoter and optionally the terminator
and/or secretory signal sequence, respectively, and to insert them
into suitable vectors containing the information necessary for
replication, are well known to persons skilled in the art (cf., for
instance, Sambrook et al., supra).
[0724] The host cell into which the DNA sequence or the recombinant
vector is introduced may be any cell which is capable of producing
the present peptide and includes bacteria, yeast, fungi and higher
eukaryotic cells. Examples of suitable host cells well known and
used in the art are, without limitation, E. coli, Saccharomyces
cerevisiae, or mammalian BHK or CHO cell lines.
[0725] Pharmaceutical compositions containing a compound according
to the present invention may be prepared by conventional
techniques, e.g. as described in Remington's Pharmaceutical
Sciences, 1985 or in Remington: The Science and Practice of
Pharmacy, 19.sup.th edition, 1995.
[0726] One object of the present invention is to provide a
pharmaceutical formulation comprising a compound according to the
present invention which is present in a concentration from about
0.1 mg/ml to about 25 mg/ml, and wherein said formulation has a pH
from 2.0 to 10.0. The formulation may further comprise a buffer
system, preservative(s), isotonicity agent(s), chelating agent(s),
stabilizers and surfactants. In one embodiment of the invention the
pharmaceutical formulation is an aqueous formulation, i.e.
formulation comprising water. Such formulation is typically a
solution or a suspension. In a further embodiment of the invention
the pharmaceutical formulation is an aqueous solution. The term
"aqueous formulation" is defined as a formulation comprising at
least 50% w/w water. Likewise, the term "aqueous solution" is
defined as a solution comprising at least 50% w/w water, and the
term "aqueous suspension" is defined as a suspension comprising at
least 50% w/w water.
[0727] In another embodiment the pharmaceutical formulation is a
freeze-dried formulation, whereto the physician or the patient adds
solvents and/or diluents prior to use.
[0728] In another embodiment the pharmaceutical formulation is a
dried formulation (e.g. freeze-dried or spray-dried) ready for use
without any prior dissolution.
[0729] In a further aspect the invention relates to a
pharmaceutical formulation comprising an aqueous solution of a
compound according to the present invention, and a buffer, wherein
said compound is present in a concentration from 0.1 mg/ml or
above, and wherein said formulation has a pH from about 2.0 to
about 10.0.
[0730] In another embodiment of the invention the pH of the
formulation is from about 7.0 to about 9.5. In another embodiment
of the invention the pH of the formulation is from about 3.0 to
about 7.0. In another embodiment of the invention the pH of the
formulation is from about 5.0 to about 7.5. In another embodiment
of the invention the pH of the formulation is from about 7.5 to
about 9.0. In another embodiment of the invention the pH of the
formulation is from about 7.5 to about 8.5. In another embodiment
of the invention the pH of the formulation is from about 6.0 to
about 7.5. In another embodiment of the invention the pH of the
formulation is from about 6.0 to about 7.0.
[0731] In another embodiment of the invention the pH of the
formulation is from about 3.0 to about 9.0, and said pH is at least
2.0 pH units from the isoelectric pH of compound of the present
invention.
[0732] In a further embodiment of the invention the buffer is
selected from the group consisting of sodium acetate, sodium
carbonate, citrate, glycylglycine, histidine, glycine, lysine,
arginin, sodium dihydrogen phosphate, disodium hydrogen phosphate,
sodium phosphate, and tris(hydroxymethyl)-aminomethan, bicine,
tricine, malic acid, succinate, maleic acid, fumaric acid, tartaric
acid, aspartic acid or mixtures thereof. Each one of these specific
buffers constitutes an alternative embodiment of the invention.
[0733] In a further embodiment of the invention the formulation
further comprises a pharmaceutically acceptable preservative. In a
further embodiment of the invention the preservative is selected
from the group consisting of phenol, o-cresol, m-cresol, p-cresol,
methyl p-hydroxybenzoate, propyl p-hydroxybenzoate,
2-phenoxyethanol, butyl p-hydroxybenzoate, 2-phenylethanol, benzyl
alcohol, chlorobutanol, and thiomerosal, bronopol, benzoic acid,
imidurea, chlorohexidine, sodium dehydroacetate, chlorocresol,
ethyl p-hydroxybenzoate, benzethonium chloride, chlorphenesine
(3p-chlorphenoxypropane-1,2-diol) or mixtures thereof.
[0734] In a further embodiment of the invention the preservative is
present in a concentration from 0.1 mg/ml to 20 mg/ml. In a further
embodiment of the invention the preservative is present in a
concentration from 0.1 mg/ml to 5 mg/ml. In a further embodiment of
the invention the preservative is present in a concentration from 5
mg/ml to 10 mg/ml. In a further embodiment of the invention the
preservative is present in a concentration from 10 mg/ml to 20
mg/ml. Each one of these specific preservatives constitutes an
alternative embodiment of the invention. The use of a preservative
in pharmaceutical compositions is well-known to the skilled person.
For convenience reference is made to Remington: The Science and
Practice of Pharmacy, 19.sup.th edition, 1995.
[0735] In a further embodiment of the invention the formulation
further comprises an isotonic agent. In a further embodiment of the
invention the isotonic agent is selected from the group consisting
of a salt (e.g. sodium chloride), a sugar or sugar alcohol, an
amino acid (e.g. L-glycine, L-histidine, arginine, lysine,
isoleucine, aspartic acid, tryptophan, threonine), an alditol (e.g.
glycerol (glycerine), 1,2-propanediol (propyleneglycol),
1,3-propanediol, 1,3-butanediol) polyethyleneglycol (e.g. PEG400),
or mixtures thereof. Any sugar such as mono-, di-, or
polysaccharides, or water-soluble glucans, including for example
fructose, glucose, mannose, sorbose, xylose, maltose, lactose,
sucrose, trehalose, dextran, pullulan, dextrin, cyclodextrin,
soluble starch, hydroxyethyl starch and carboxymethylcellulose-Na
may be used. In one embodiment the sugar additive is sucrose. Sugar
alcohol is defined as a C4-C8 hydrocarbon having at least one --OH
group and includes, for example, mannitol, sorbitol, inositol,
galacititol, dulcitol, xylitol, and arabitol. In one embodiment the
sugar alcohol additive is mannitol. The sugars or sugar alcohols
mentioned above may be used individually or in combination. There
is no fixed limit to the amount used, as long as the sugar or sugar
alcohol is soluble in the liquid preparation and does not adversely
effect the stabilizing effects achieved using the methods of the
invention. In one embodiment, the sugar or sugar alcohol
concentration is between about 1 mg/ml and about 150 mg/ml. In a
further embodiment of the invention the isotonic agent is present
in a concentration from 1 mg/ml to 50 mg/ml. In a further
embodiment of the invention the isotonic agent is present in a
concentration from 1 mg/ml to 7 mg/ml. In a further embodiment of
the invention the isotonic agent is present in a concentration from
8 mg/ml to 24 mg/ml. In a further embodiment of the invention the
isotonic agent is present in a concentration from 25 mg/ml to 50
mg/ml. Each one of these specific isotonic agents constitutes an
alternative embodiment of the invention. The use of an isotonic
agent in pharmaceutical compositions is well-known to the skilled
person. For convenience reference is made to Remington: The Science
and Practice of Pharmacy, 19.sup.th edition, 1995.
[0736] In a further embodiment of the invention the formulation
further comprises a chelating agent. In a further embodiment of the
invention the chelating agent is selected from salts of
ethylenediaminetetraacetic acid (EDTA), citric acid, and aspartic
acid, and mixtures thereof. In a further embodiment of the
invention the chelating agent is present in a concentration from
0.1 mg/ml to 5 mg/ml. In a further embodiment of the invention the
chelating agent is present in a concentration from 0.1 mg/ml to 2
mg/ml. In a further embodiment of the invention the chelating agent
is present in a concentration from 2 mg/ml to 5 mg/ml. Each one of
these specific chelating agents constitutes an alternative
embodiment of the invention. The use of a chelating agent in
pharmaceutical compositions is well-known to the skilled person.
For convenience reference is made to Remington: The Science and
Practice of Pharmacy, 19.sup.th edition, 1995.
[0737] In a further embodiment of the invention the formulation
further comprises a stabiliser. The use of a stabilizer in
pharmaceutical compositions is well-known to the skilled person.
For convenience reference is made to Remington: The Science and
Practice of Pharmacy, 19.sup.th edition, 1995.
[0738] More particularly, compositions of the invention are
stabilized liquid pharmaceutical compositions whose therapeutically
active components include a polypeptide that possibly exhibits
aggregate formation during storage in liquid pharmaceutical
formulations. By "aggregate formation" is intended a physical
interaction between the polypeptide molecules that results in
formation of oligomers, which may remain soluble, or large visible
aggregates that precipitate from the solution. By "during storage"
is intended a liquid pharmaceutical composition or formulation once
prepared, is not immediately administered to a subject. Rather,
following preparation, it is packaged for storage, either in a
liquid form, in a frozen state, or in a dried form for later
reconstitution into a liquid form or other form suitable for
administration to a subject. By "dried form" is intended the liquid
pharmaceutical composition or formulation is dried either by freeze
drying (i.e., lyophilization; see, for example, Williams and Polli
(1984) J. Parenteral Sci. Technol. 38:48-59), spray drying (see
Masters (1991) in Spray-Drying Handbook (5th ed; Longman Scientific
and Technical, Essez, U.K.), pp. 491-676; Broadhead et al. (1992)
Drug Devel. Ind. Pharm. 18:1169-1206; and Mumenthaler et al. (1994)
Pharm. Res. 11:12-20), or air drying (Carpenter and Crowe (1988)
Cryobiology 25:459-470; and Roser (1991) Biopharm. 4:47-53).
Aggregate formation by a polypeptide during storage of a liquid
pharmaceutical composition can adversely affect biological activity
of that polypeptide, resulting in loss of therapeutic efficacy of
the pharmaceutical composition. Furthermore, aggregate formation
may cause other problems such as blockage of tubing, membranes, or
pumps when the polypeptide-containing pharmaceutical composition is
administered using an infusion system.
[0739] The pharmaceutical compositions of the invention may further
comprise an amount of an amino acid base sufficient to decrease
aggregate formation by the polypeptide during storage of the
composition. By "amino acid base" is intended an amino acid or a
combination of amino acids, where any given amino acid is present
either in its free base form or in its salt form. Where a
combination of amino acids is used, all of the amino acids may be
present in their free base forms, all may be present in their salt
forms, or some may be present in their free base forms while others
are present in their salt forms. In one embodiment, amino acids to
use in preparing the compositions of the invention are those
carrying a charged side chain, such as arginine, lysine, aspartic
acid, and glutamic acid. Any stereoisomer (i.e., L, D, or DL
isomer) of a particular amino acid (e.g. glycine, methionine,
histidine, imidazole, arginine, lysine, isoleucine, aspartic acid,
tryptophan, threonine and mixtures thereof) or combinations of
these stereoisomers, may be present in the pharmaceutical
compositions of the invention so long as the particular amino acid
is present either in its free base form or its salt form. In one
embodiment the L-stereoisomer is used. Compositions of the
invention may also be formulated with analogues of these amino
acids. By "amino acid analogue" is intended a derivative of the
naturally occurring amino acid that brings about the desired effect
of decreasing aggregate formation by the polypeptide during storage
of the liquid pharmaceutical compositions of the invention.
Suitable arginine analogues include, for example, aminoguanidine,
ornithine and N-monoethyl L-arginine, suitable methionine analogues
include S-ethyl homocysteine and S-butyl homocysteine and suitable
cystein analogues include S-methyl-L cystein. As with the other
amino acids, the amino acid analogues are incorporated into the
compositions in either their free base form or their salt form. In
a further embodiment of the invention the amino acids or amino acid
analogues are used in a concentration, which is sufficient to
prevent or delay aggregation of the protein.
[0740] In a further embodiment of the invention methionine (or
other sulphur containing amino acids or amino acid analogous) may
be added to inhibit oxidation of methionine residues to methionine
sulfoxide when the polypeptide acting as the therapeutic agent is a
polypeptide comprising at least one methionine residue susceptible
to such oxidation. By "inhibit" is intended minimal accumulation of
methionine oxidized species over time. Inhibiting methionine
oxidation results in greater retention of the polypeptide in its
proper molecular form. Any stereoisomer of methionine (L, D, or DL
isomer) or combinations thereof can be used. The amount to be added
should be an amount sufficient to inhibit oxidation of the
methionine residues such that the amount of methionine sulfoxide is
acceptable to regulatory agencies. Typically, this means that the
composition contains no more than about 10% to about 30% methionine
sulfoxide. Generally, this can be achieved by adding methionine
such that the ratio of methionine added to methionine residues
ranges from about 1:1 to about 1000:1, such as 10:1 to about
100:1.
[0741] In a further embodiment of the invention the formulation
further comprises a stabiliser selected from the group of high
molecular weight polymers or low molecular compounds. In a further
embodiment of the invention the stabilizer is selected from
polyethylene glycol (e.g. PEG 3350), polyvinylalcohol (PVA),
polyvinylpyrrolidone, carboxy-/hydroxycellulose or derivates
thereof (e.g. HPC, HPC-SL, HPC-L and HPMC), cyclodextrins,
sulphur-containing substances as monothioglycerol, thioglycolic
acid and 2-methylthioethanol, and different salts (e.g. sodium
chloride). Each one of these specific stabilizers constitutes an
alternative embodiment of the invention.
[0742] The pharmaceutical compositions may also comprise additional
stabilizing agents, which further enhance stability of a
therapeutically active polypeptide therein. Stabilizing agents of
particular interest to the present invention include, but are not
limited to, methionine and EDTA, which protect the polypeptide
against methionine oxidation, and a nonionic surfactant, which
protects the polypeptide against aggregation associated with
freeze-thawing or mechanical shearing.
[0743] In a further embodiment of the invention the formulation
further comprises a surfactant. In a further embodiment of the
invention the surfactant is selected from a detergent, ethoxylated
castor oil, polyglycolyzed glycerides, acetylated monoglycerides,
sorbitan fatty acid esters, polyoxypropylene-polyoxyethylene block
polymers (eg. poloxamers such as Pluronic.RTM. F68, poloxamer 188
and 407, Triton X-100), polyoxyethylene sorbitan fatty acid esters,
polyoxyethylene and polyethylene derivatives such as alkylated and
alkoxylated derivatives (tweens, e.g. Tween-20, Tween-40, Tween-80
and Brij-35), monoglycerides or ethoxylated derivatives thereof,
diglycerides or polyoxyethylene derivatives thereof, alcohols,
glycerol, lecitins and phospholipids (eg. phosphatidyl serine,
phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl
inositol, diphosphatidyl glycerol and sphingomyelin), derivates of
phospholipids (eg. dipalmitoyl phosphatidic acid) and
lysophospholipids (eg. palmitoyl lysophosphatidyl-L-serine and
1-acyl-sn-glycero-3-phosphate esters of ethanolamine, choline,
serine or threonine) and alkyl, alkoxyl (alkyl ester), alkoxy
(alkyl ether)-derivatives of lysophosphatidyl and
phosphatidylcholines, e.g. lauroyl and myristoyl derivatives of
lysophosphatidylcholine, dipalmitoylphosphatidylcholine, and
modifications of the polar head group, that is cholines,
ethanolamines, phosphatidic-acid, serines, threonines, glycerol,
inositol, and the positively charged DODAC, DOTMA, DCP, BISHOP,
lysophosphatidylserine and lysophosphatidylthreonine, and
glycerophospholipids (eg. cephalins), glyceroglycolipids (eg.
galactopyransoide), sphingoglycolipids (eg. ceramides,
gangliosides), dodecylphosphocholine, hen egg lysolecithin, fusidic
acid derivatives--(e.g. sodium tauro-dihydrofusidate etc.),
long-chain fatty acids and salts thereof C6-C12 (eg. oleic acid and
caprylic acid), acylcarnitines and derivatives,
N.sup..alpha.-acylated derivatives of lysine, arginine or
histidine, or side-chain acylated derivatives of lysine or
arginine, N.sup..alpha.-acylated derivatives of dipeptides
comprising any combination of lysine, arginine or histidine and a
neutral or acidic amino acid, N.sup..alpha.-acylated derivative of
a tripeptide comprising any combination of a neutral amino acid and
two charged amino acids, DSS (docusate sodium, CAS registry no
[577-11-7]), docusate calcium, CAS registry no [128-49-4]),
docusate potassium, CAS registry no [7491-09-0]), SDS (sodium
dodecyl sulfate or sodium lauryl sulfate), sodium caprylate, cholic
acid or derivatives thereof, bile acids and salts thereof and
glycine or taurine conjugates, ursodeoxycholic acid, sodium
cholate, sodium deoxycholate, sodium taurocholate, sodium
glycocholate,
N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate, anionic
(alkyl-aryl-sulphonates) monovalent surfactants, zwitterionic
surfactants (e.g. N-alkyl-N,N-dimethylammonio-1-propanesulfonates,
3-cholamido-1-propyldimethylammonio-1-propanesulfonate, cationic
surfactants (quarternary ammonium bases) (e.g.
cetyl-trimethylammonium bromide, cetylpyridinium chloride),
non-ionic surfactants (eg. dodecyl .beta.-D-glucopyranoside),
poloxamines (eg. Tetronic's), which are tetrafunctional block
copolymers derived from sequential addition of propylene oxide and
ethylene oxide to ethylenediamine, or the surfactant may be
selected from the group of imidazoline derivatives, or mixtures
thereof. Each one of these specific surfactants constitutes an
alternative embodiment of the invention.
[0744] The use of a surfactant in pharmaceutical compositions is
well-known to the skilled person. For convenience reference is made
to Remington: The Science and Practice of Pharmacy, 19.sup.th
edition, 1995.
[0745] It is possible that other ingredients may be present in the
peptide pharmaceutical formulation of the present invention. Such
additional ingredients may include wetting agents, emulsifiers,
antioxidants, bulking agents, tonicity modifiers, chelating agents,
metal ions, oleaginous vehicles, proteins (e.g., human serum
albumin, gelatin or proteins) and a zwitterion (e.g., an amino acid
such as betaine, taurine, arginine, glycine, lysine and histidine).
Such additional ingredients, of course, should not adversely affect
the overall stability of the pharmaceutical formulation of the
present invention.
[0746] Pharmaceutical compositions containing a compound according
to the present invention may be administered to a patient in need
of such treatment at several sites, for example, at topical sites,
for example, skin and mucosal sites, at sites which bypass
absorption, for example, administration in an artery, in a vein, in
the heart, and at sites which involve absorption, for example,
administration in the skin, under the skin, in a muscle or in the
abdomen.
[0747] Administration of pharmaceutical compositions according to
the invention may be through several routes of administration, for
example, lingual, sublingual, buccal, in the mouth, oral, in the
stomach and intestine, nasal, pulmonary, for example, through the
bronchioles and alveoli or a combination thereof, epidermal,
dermal, transdermal, vaginal, rectal, ocular, for examples through
the conjunctiva, uretal, and parenteral to patients in need of such
a treatment.
[0748] In one aspect the present invention relates to a
pharmaceutical composition comprising a compound according to
Formula (I), and a pharmaceutically acceptable excipient.
[0749] In one embodiment the pharmaceutical composition is suited
for pulmonary administration.
[0750] In another aspect the present invention relates to the use
of a compound of formula (I) for the preparation of a pulmonary
medicament.
[0751] Compositions of the current invention may be administered in
several dosage forms, for example, as solutions, suspensions,
emulsions, microemulsions, multiple emulsion, foams, salves,
pastes, plasters, ointments, tablets, coated tablets, rinses,
capsules, for example, hard gelatine capsules and soft gelatine
capsules, suppositories, rectal capsules, drops, gels, sprays,
powder, aerosols, inhalants, eye drops, ophthalmic ointments,
ophthalmic rinses; vaginal pessaries, vaginal rings, vaginal
ointments, injection solution, in situ transforming solutions, for
example in situ gelling, in situ setting, in situ precipitating, in
situ crystallization, infusion solution, and implants.
[0752] Compositions of the invention may further be compounded in,
or attached to, for example through covalent, hydrophobic and
electrostatic interactions, a drug carrier, drug delivery system
and advanced drug delivery system in order to further enhance
stability of the compound, increase bioavailability, increase
solubility, decrease adverse effects, achieve chrono-therapy well
known to those skilled in the art, and increase patient compliance
or any combination thereof. Examples of carriers, drug delivery
systems and advanced drug delivery systems include, but are not
limited to, polymers, for example cellulose and derivatives,
polysaccharides, for example dextran and derivatives, starch and
derivatives, poly(vinyl alcohol), acrylate and methacrylate
polymers, polylactic and polyglycolic acid and block co-polymers
thereof, polyethylene glycols, carrier proteins, for example
albumin, gels, for example, thermogelling systems, for example
block co-polymeric systems well known to those skilled in the art,
micelles, liposomes, microspheres, nanoparticulates, liquid
crystals and dispersions thereof, L2 phase and dispersions there
of, well known to those skilled in the art of phase behaviour in
lipid-water systems, polymeric micelles, multiple emulsions,
self-emulsifying, self-microemulsifying, cyclodextrins and
derivatives thereof, and dendrimers.
[0753] Compositions of the current invention are useful in the
formulation of solids, semisolids, powder and solutions for
pulmonary administration of the compound, using, for example a
metered dose inhaler, dry powder inhaler and a nebulizer, all being
devices well known to those skilled in the art.
[0754] Compositions of the current invention are specifically
useful in the formulation of controlled, sustained, protracting,
retarded, and slow release drug delivery systems. More
specifically, but not limited to, compositions are useful in
formulation of parenteral controlled release and sustained release
systems (both systems leading to a many-fold reduction in number of
administrations), well known to those skilled in the art. Even more
preferably, are controlled release and sustained release systems
administered subcutaneous. Without limiting the scope of the
invention, examples of useful controlled release system and
compositions are hydrogels, oleaginous gels, liquid crystals,
polymeric micelles, microspheres, nanoparticles,
[0755] Methods to produce controlled release systems useful for
compositions of the current invention include, but are not limited
to, crystallization, condensation, co-cystallization,
precipitation, co-precipitation, emulsification, dispersion, high
pressure homogenization, encapsulation, spray drying,
microencapsulation, coacervation, phase separation, solvent
evaporation to produce microspheres, extrusion and supercritical
fluid processes. General reference is made to Handbook of
Pharmaceutical Controlled Release (Wise, D. L., ed. Marcel Dekker,
New York, 2000) and Drug and the Pharmaceutical Sciences vol. 99:
Protein Formulation and Delivery (MacNally, E. J., ed. Marcel
Dekker, New York, 2000).
[0756] Parenteral administration may be performed by subcutaneous,
intramuscular, intraperitoneal or intravenous injection by means of
a syringe, optionally a pen-like syringe. Alternatively, parenteral
administration can be performed by means of an infusion pump. A
further option is a composition which may be a solution or
suspension for the administration of the compound according to the
present invention in the form of a nasal or pulmonal spray. As a
still further option, the pharmaceutical compositions containing
the compound of the invention can also be adapted to transdermal
administration, e.g. by needle-free injection or from a patch,
optionally an iontophoretic patch, or transmucosal, e.g. buccal,
administration.
[0757] The term "stabilized formulation" refers to a formulation
with increased physical stability, increased chemical stability or
increased physical and chemical stability.
[0758] The term "physical stability" of the protein formulation as
used herein refers to the tendency of the protein to form
biologically inactive and/or insoluble aggregates of the protein as
a result of exposure of the protein to thermo-mechanical stresses
and/or interaction with interfaces and surfaces that are
destabilizing, such as hydrophobic surfaces and interfaces.
Physical stability of the aqueous protein formulations is evaluated
by means of visual inspection and/or turbidity measurements after
exposing the formulation filled in suitable containers (e.g.
cartridges or vials) to mechanical/physical stress (e.g. agitation)
at different temperatures for various time periods. Visual
inspection of the formulations is performed in a sharp focused
light with a dark background. The turbidity of the formulation is
characterized by a visual score ranking the degree of turbidity for
instance on a scale from 0 to 3 (a formulation showing no turbidity
corresponds to a visual score 0, and a formulation showing visual
turbidity in daylight corresponds to visual score 3). A formulation
is classified physical unstable with respect to protein
aggregation, when it shows visual turbidity in daylight.
Alternatively, the turbidity of the formulation can be evaluated by
simple turbidity measurements well-known to the skilled person.
Physical stability of the aqueous protein formulations can also be
evaluated by using a spectroscopic agent or probe of the
conformational status of the protein. The probe is preferably a
small molecule that preferentially binds to a non-native conformer
of the protein. One example of a small molecular spectroscopic
probe of protein structure is Thioflavin T. Thioflavin T is a
fluorescent dye that has been widely used for the detection of
amyloid fibrils. In the presence of fibrils, and perhaps other
protein configurations as well, Thioflavin T gives rise to a new
excitation maximum at about 450 nm and enhanced emission at about
482 nm when bound to a fibril protein form. Unbound Thioflavin T is
essentially non-fluorescent at the wavelengths.
[0759] Other small molecules can be used as probes of the changes
in protein structure from native to non-native states. For instance
the "hydrophobic patch" probes that bind preferentially to exposed
hydrophobic patches of a protein. The hydrophobic patches are
generally buried within the tertiary structure of a protein in its
native state, but become exposed as a protein begins to unfold or
denature. Examples of these small molecular, spectroscopic probes
are aromatic, hydrophobic dyes, such as antrhacene, acridine,
phenanthroline or the like. Other spectroscopic probes are
metal-amino acid complexes, such as cobalt metal complexes of
hydrophobic amino acids, such as phenylalanine, leucine,
isoleucine, methionine, and valine, or the like.
[0760] The term "chemical stability" of the protein formulation as
used herein refers to chemical covalent changes in the protein
structure leading to formation of chemical degradation products
with potential less biological potency and/or potential increased
immunogenic properties compared to the native protein structure.
Various chemical degradation products can be formed depending on
the type and nature of the native protein and the environment to
which the protein is exposed. Elimination of chemical degradation
can most probably not be completely avoided and increasing amounts
of chemical degradation products is often seen during storage and
use of the protein formulation as well-known by the person skilled
in the art. Most proteins are prone to deamidation, a process in
which the side chain amide group in glutaminyl or asparaginyl
residues is hydrolysed to form a free carboxylic acid. Other
degradations pathways involves formation of high molecular weight
transformation products where two or more protein molecules are
covalently bound to each other through transamidation and/or
disulfide interactions leading to formation of covalently bound
dimer, oligomer and polymer degradation products (Stability of
Protein Pharmaceuticals, Ahern. T. J. & Manning M. C., Plenum
Press, New York 1992). Oxidation (of for instance methionine
residues) can be mentioned as another variant of chemical
degradation. The chemical stability of the protein formulation can
be evaluated by measuring the amount of the chemical degradation
products at various time-points after exposure to different
environmental conditions (the formation of degradation products can
often be accelerated by for instance increasing temperature). The
amount of each individual degradation product is often determined
by separation of the degradation products depending on molecule
size and/or charge using various chromatography techniques (e.g.
SEC-HPLC and/or RP-HPLC).
[0761] Hence, as outlined above, a "stabilized formulation" refers
to a formulation with increased physical stability, increased
chemical stability or increased physical and chemical stability. In
general, a formulation must be stable during use and storage (in
compliance with recommended use and storage conditions) until the
expiration date is reached.
[0762] In one embodiment of the invention the pharmaceutical
formulation comprising the compound according to the present
invention is stable for more than 6 weeks of usage and for more
than 3 years of storage.
[0763] In another embodiment of the invention the pharmaceutical
formulation comprising the compound according to the present
invention is stable for more than 4 weeks of usage and for more
than 3 years of storage.
[0764] In a further embodiment of the invention the pharmaceutical
formulation comprising the compound according to the present
invention is stable for more than 4 weeks of usage and for more
than two years of storage.
[0765] In an even further embodiment of the invention the
pharmaceutical formulation comprising the compound is stable for
more than 2 weeks of usage and for more than two years of
storage.
[0766] In another aspect the present invention relates to the use
of a compound according to the invention for the preparation of a
medicament.
[0767] In one embodiment a compound according to the invention is
used for the preparation of a medicament for the treatment or
prevention of hyperglycemia, type 2 diabetes, impaired glucose
tolerance, type 1 diabetes, obesity, hypertension, syndrome X,
dyslipidemia, cognitive disorders, atheroschlerosis, myocardial
infarction, coronary heart disease and other cardiovascular
disorders, stroke, inflammatory bowel syndrome, dyspepsia and
gastric ulcers.
[0768] In another embodiment a compound according to the invention
is used for the preparation of a medicament for delaying or
preventing disease progression in type 2 diabetes.
[0769] In another embodiment a compound according to the invention
is used for the preparation of a medicament for decreasing food
intake, decreasing .beta.-cell apoptosis, increasing .beta.-cell
function and .beta.-cell mass, and/or for restoring glucose
sensitivity to .beta.-cells.
[0770] The treatment with a compound according to the present
invention may also be combined with combined with a second or more
pharmacologically active substances, e.g. selected from
antidiabetic agents, antiobesity agents, appetite regulating
agents, antihypertensive agents, agents for the treatment and/or
prevention of complications resulting from or associated with
diabetes and agents for the treatment and/or prevention of
complications and disorders resulting from or associated with
obesity. Examples of these pharmacologically active substances are:
Insulin, sulphonylureas, biguanides, meglitinides, glucosidase
inhibitors, glucagon antagonists, DPP-IV (dipeptidyl peptidase-IV)
inhibitors, inhibitors of hepatic enzymes involved in stimulation
of gluconeogenesis and/or glycogenolysis, glucose uptake
modulators, compounds modifying the lipid metabolism such as
antihyperlipidemic agents as HMG CoA inhibitors (statins),
compounds lowering food intake, RXR agonists and agents acting on
the ATP-dependent potassium channel of the .beta.-cells;
Cholestyramine, colestipol, clofibrate, gemfibrozil, lovastatin,
pravastatin, simvastatin, probucol, dextrothyroxine, neteglinide,
repaglinide; .beta.-blockers such as alprenolol, atenolol, timolol,
pindolol, propranolol and metoprolol, ACE (angiotensin converting
enzyme) inhibitors such as benazepril, captopril, enalapril,
fosinopril, lisinopril, alatriopril, quinapril and ramipril,
calcium channel blockers such as nifedipine, felodipine,
nicardipine, isradipine, nimodipine, diltiazem and verapamil, and
.alpha.-blockers such as doxazosin, urapidil, prazosin and
terazosin; CART (cocaine amphetamine regulated transcript)
agonists, NPY (neuropeptide Y) antagonists, MC4 (melanocortin 4)
agonists, orexin antagonists, TNF (tumor necrosis factor) agonists,
CRF (corticotropin releasing factor) agonists, CRF BP
(corticotropin releasing factor binding protein) antagonists,
urocortin agonists, .beta.3 agonists, MSH (melanocyte-stimulating
hormone) agonists, MCH (melanocyte-concentrating hormone)
antagonists, CCK (cholecystokinin) agonists, serotonin re-uptake
inhibitors, serotonin and noradrenaline re-uptake inhibitors, mixed
serotonin and noradrenergic compounds, 5HT (serotonin) agonists,
bombesin agonists, galanin antagonists, growth hormone, growth
hormone releasing compounds, TRH (thyreotropin releasing hormone)
agonists, UCP 2 or 3 (uncoupling protein 2 or 3) modulators, leptin
agonists, DA agonists (bromocriptin, doprexin), lipase/amylase
inhibitors, RXR (retinoid X receptor) modulators, TR .beta.
agonists; histamine H3 antagonists.
[0771] It should be understood that any suitable combination of the
compounds according to the invention with one or more of the
above-mentioned compounds and optionally one or more further
pharmacologically active substances are considered to be within the
scope of the present invention.
[0772] The present invention is further illustrated by the
following examples which, however, are not to be construed as
limiting the scope of protection. The features disclosed in the
foregoing description and in the following examples may, both
separately and in any combination thereof, be material for
realising the invention in diverse forms thereof.
EXAMPLES
Abbreviations used:
r.t retention time
TFE trifluoroethanol
DIEA diisopropylethylamine
H.sub.2O water
CH.sub.3CN acetonitrile
DMF NN dimethylformamide
HBTU 2-(1H-Benzotriazol-1-yl-)-1,1,3,3 tetramethyluronium
hexafluorophosphate
HATU O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-Tetramethyluronium
Hexafluorophosphate
ImPr 3-(1-Imidazol-4-yl)-propionyl
Adoc 1-Adamantyloxycarbonyl
Fmoc 9H-fluoren-9-ylmethoxycarbonyl
Boc tert butyloxycarbonyl
OtBu tert butyl ester
tBu tert butyl
Trt triphenylmethyl
Pmc 2,2,5,7,8-Pentamethyl-chroman-6-sulfonyl
Dde 1-(4,4-Dimethyl-2,6-dioxocyclohexylidene)ethyl
ivDde 1-(4,4-Dimethyl-2,6-dioxocyclohexylidene)-3-Methylbutyl
DCM dichloromethane
TIS triisopropylsilane)
TFA: trifluoroacetic acid
Et.sub.2O: diethylether
NMP 1-Methyl-pyrrolidin-2-one
Aib .alpha.-aminoisobutyric acid
Analysis
[0773] HPLC analysis by the methods A1, B1 and B6 was performed on
a Waters 2690 Separation Module equipped with a Waters 996 diode
array detector. A Vydac 218TP54 4.6 mm.times.250 mm 5 .mu.m C-18
silica column (The Separations Group, Hesperia) was used and
detection was by UV at 214 nm, 254 nm, 280 nm and 301 nm.
[0774] HPLC analysis by the method 01_B4.sub.--2 was performed on a
Waters 600S system fitted with a Waters 996 diode array detector. A
Symmetry300 C18, 5 .mu.m, 3.9 mm.times.150 mm column (Waters) was
used and detection was by UV at 214 nm and 254 nm.
[0775] In method A1 the column was equilibrated with 0.05
M.NH.sub.4SO.sub.4 pH 3.5 and eluted by a gradient of 0 to 60%
CH.sub.3CN in 0.05 M (NH.sub.4).sub.2SO.sub.4 pH 3.5 over 50 min at
42.degree. C., with a flow of 0.5 ml/min.
[0776] In method B1 the column was equilibrated with 0.1%
TFA/H.sub.2O and eluted by a gradient of 0 to 60% CH.sub.3CN
against 0.1% TFA/H.sub.2O over 50 min at 42.degree. C., with a flow
of 0.5 ml/min.
[0777] In method B6 the column was equilibrated with 0.1%
TFA/H.sub.2O and eluted by a gradient of 0 to 90% CH.sub.3CN
against 0.1% TFA/H.sub.2O over 50 min at 42.degree. C., with a flow
of 0.5 ml/min.
[0778] In method 01_B4.sub.--2 the column was equilibrated with 5%
acetonitrile in water with 0.05% TFA and eluted by a gradient of 5
to 65% CH.sub.3CN against 0.05% TFA/H.sub.2O over 15 min at
42.degree. C., with a flow of 1 ml/min.
[0779] Protein amount was calculated by comparing the UV detector
response of the sample with the detector response from at of from a
hGH standard for which the amount has been determined by amino acid
analysis.
[0780] LC-MS analysis was performed on a PE-Sciex API 100 mass
spectrometer equipped with two Perkin Elmer Series 200 Micropumps,
a Perkin Elmer Series 200 autosampler, a Applied Biosystems 785A UV
detector and a Sedex 75 Evaporative Light scattering detector. A
Waters Xterra 3.0 mm.times.50 mm 5.mu. C-18 silica column was
eluted at 1.5 ml/min at room temperature. It was equilibrated with
5% CH.sub.3CN/0.1% TFA/H.sub.2O and eluted for 1.0 min with 5%
CH.sub.3CN/0.1% TFA/H.sub.2O and then with a linear gradient to 90%
CH.sub.3CN/0.1% TFA/H.sub.2O over 7 min. Detection was by UV
detection at 214 nm and Evaporative light Scattering. A fraction of
the column eluate was introduced into the ionspray interface of a
PE-Sciex API 100 mass spectrometer. The mass range 300-2000 amu was
scanned every 2 seconds during the run. Maldi TOF MS analysis was
performed on a Bruker Autoflex instrument in linear mode. Samples a
prepared by the thin layer dried droplet method using
.alpha.-cyano-4-hydroxycinnamic acid as the matrix.
Example 1
Preparation of
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(-
7-37) amide wherein mPEGyl is Polydisperse and has a Molecular
Weight of Approximately 2 kDa
[0781] ##STR15## 1.a Synthesis of the Protected Peptidyl Resin.
[0782]
Boc-His(Boc)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(-
OtBu)-Val-Ser(tBu)Ser(tBu)-Tyr(tBu)-Leu-Glu(OtBu)-Aib-Gln(Trt)-Ala-Ala-Lys-
(Boc)-Glu(OtBu)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Lys(Boc)-Aib-Arg(Pmc)-Lys(Dde-
)-Rink amide resin was prepared according to the Fmoc strategy on
an Applied Biosystems 433A peptide synthesizer in 0.25 mmol scale
using the manufacturer supplied FastMoc UV protocols which employ
HBTU mediated couplings in NMP, and UV monitoring of the
deprotection of the Fmoc protection group. To improve the coupling
efficiency, Aib residues and residues following Aib, these residues
were coupled using HATU instead of HBTU as the coupling reagent.
The starting resin (438 mg) used for the synthesis was
4-(2',4'-Dimethoxyphenyl-Fmoc-aminomethyl)-phenoxy resin (Rink
amide resin) (Merck Biosciences GmbH, Germany. cat. #: 01-12-0013)
with a substitution capacity of 0.57 mmol/g. The protected amino
acid derivatives used were
(2S)-6-[1-(4,4-Dimethyl-2,6-dioxocyclohexylidene)-ethylamino]-2-(9H-fluor-
en-9-ylmethoxycarbonylamino)-hexanoic acid (Fmoc-Lys(Dde)-OH),
Fmoc-Arg(Pmc)-OH, Fmoc-Aib-OH, Fmoc-Lys(Boc)-OH, Fmoc-Val-OH,
Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Ala-OH, Fmoc-Ile-OH,
Fmoc-Phe-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Tyr(tBu)-OH,
Fmoc-Ser(tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Gly-OH
and Boc-His(Boc)-OH
[0783] The yield was 1.37 g of dry peptidyl resin.
1.b Characterisation of the Peptidyl Resin
[0784] The resin was characterized by cleaving off the crude
peptide from 50 mg of this resin by treating it for 2 hours with a
mixture of 14 .mu.l TIS, 14 .mu.l H.sub.2O and 0.5 ml TFA. The
resin was removed by filtration and the crude peptide was isolated
by precipitation and wash with Et.sub.2O. HPLC and LC-MS analysis
was performed on the dry precipitate.
[0785] Analytical Results: TABLE-US-00001 Analytical method Result
HPLC A1 r.t.: 37.41 min., LC-MS r.t. 3.48 min., Mass for (M +
3H.sup.+)/3: 1221.3 Da, (calc.: 1220 Da)
1.c Deprotection of Dde
[0786] The protected peptidyl resin resulting from (1.a) (1.35 g,
250 .mu.mol) was washed in NMP:DCM 1:1 (15 ml) twice. A freshly
prepared solution of hydrazine hydrate 2% in NMP (20 ml) was added.
The reaction mixture was shaken for 12 min at room temperature, and
then filtered. The hydrazine treatment was repeated twice. After
this the resin was washed extensively with NMP, DCM and NMP.
1.d Pegylation
[0787] The Dde deprotected resin was suspended in NMP (20 ml).
3-(mPEGyl)propionic acid 2,5-dioxo-pyrrolidin-1-yl ester (2.0 g, 1
mmol, 4 eq.) and DIEA (344 .mu.l, 2 mmol, 8 eq.) was added and the
suspension was shaken overnight. Then the resin was isolated by
filtration and washed extensively with NMP, DCM, 2-propanol,
methanol and Et.sub.2O and dried in vacuo.
1.e Cleavage of the Product
[0788] The resin from 1.d was stirred for 3 h at room temperature
with a mixture of 350 .mu.l TIS, 350 .mu.l H.sub.2O and 14 ml TFA.
The resin was removed by filtration and washed with 3 ml TFA. The
collected filtrates were concentrated in vacuo. to 5 ml and the
crude product was precipitated by addition of 40 ml Et.sub.2O
followed by centrifugation. The pellet was washed with 40 ml
Et.sub.2O two times and then air dried.
[0789] Results from HPLC of the dry precipitate: TABLE-US-00002
Analytical method Result HPLC A1 r.t.: 36.15 min., HPLC B6 r.t.:
28.573 min. estimated purity: 64%
1.f Purification of Product.
[0790] The crude peptide was dissolved in H.sub.2O/AcOH (40:4) (40
ml) and purified by semipreparative HPLC in 2 runs on a 25
mm.times.250 mm column packed with 7.mu. C-18 silica. The column
was eluted with a gradient of CH.sub.3CN from 40 to 62% against
0.1% TFA/H.sub.2O at 10 ml/min at a temperature of 40.degree. C.
for 47 min. The peptide containing fractions are collected, diluted
with 3 volumes of H.sub.2O and lyophilized. The final product
obtained was characterized by HPLC. TABLE-US-00003 Analytical
method Result HPLC A1 r.t.: 36.15 min.,
Example 2
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(7-
-37) wherein mPEGyl is Polydisperse and has a Molecular Weight of
Approximately 5 kDa
[0791] ##STR16## 2.a Synthesis of the Protected Peptidyl Resin
[0792] The protected peptidyl resin
Boc-His(Boc)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)Asp(OtBu)-Va-
l-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Leu-Glu(OtBu)-Aib-Gln(Trt)-Ala-Ala-Lys(Boc)Gl-
u(OtBu)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Lys(Boc)-Aib-Arg(Pmc)-Lys(Dde)-2-Chlo-
rotrityl resin was synthesized using the procedures in example 1.a.
with the exception that the starting resin was 337 mg of
(2S)-6-[1-(4,4-Dimethyl-2,6-dioxocyclohexylidene)ethylamino]-2-(((9H-fluo-
ren-9-yl)methoxycarbonyl)amino)hexanoyl 2-Chlorotrityl resin
(Fmoc-Lys(Dde)-2-ClTrt resin). This Fmoc-Lys(Dde)-2-ClTrt resin was
prepared by suspending 1 g of 2-Chlorotrityl chloride resin
(Bachem, Switzerland. cat. #: D-1965), having a substitution
capacity of 1.15 mmol/g in a mixture of 10 ml DCM and 100 .mu.l
DMF. To this was added 533 mg Fmoc-Lys(Dde)-OH (Merck Biosciences,
Germany cat. # 04-12-1121) and 684 .mu.l DIEA and the mixture was
stirred for 2 hours. The resin was isolated by filtration and then
washed three times with 10 ml of DCM/MeOH/DIEA 17:2:1, three times
with 10 ml DCM, two times with 10 ml NMP, two times with 10 ml DCM
and then finally dried in vacuo. The synthesis resulted in
approximately 1.2 g of protected peptidyl resin (air dried).
2.b Deprotection of Dde and Cleavage of Protected Peptide.
[0793] 1.0 g of the protected peptidyl resin from 2.a was treated
with 25 ml 2% hydrazine hydrate in DMF for 25 min and the resin was
isolated by filtration. This was repeated further two times and
after this the resin was thoroughly washed sequentially with DMF,
DCM, 2-propanol, methanol Et.sub.2O ether and then dried in
vacuo.
[0794] This resin which was characterized as in example 1.b.
Analytical results: TABLE-US-00004 Analytical method Result HPLC A1
r.t.: 41.12 min., HPLC B6 r.t.: 29.89 min. LC-MS r.t. 3.18 min.,
Mass for (M + 3H.sup.+)/3: 1166.6 Da, (calc.: 1166.6 Da)
[0795] The dry resin was stirred with 25 ml of a mixture of Acetic
acid/TFE/DCM 1:1:3 for 2 h and then filtered and washed thoroughly
with further 25 ml of this mixture. The pooled filtrates were
concentrated to an oil in vacuo. and the oil was stripped 5 times
with heptane to remove residual acetic acid.
2.c Pegylation and Final Deprotection.
[0796] To 100 mg of crude protected peptide from 2.b was dissolved
in 1 ml TFE at 45.degree. C. and 8.5 .mu.l DIEA was added. A
solution of 100 mg mPEG-5000-SPA (mPEG-SPA m.w. 5.000 Lot.
PT-09B-12, Shearwater, Ala., USA) in 100 .mu.l H.sub.2O and 900
.mu.l NMP was added and the mixture was stirred overnight at r.t.
After this 48 ml Et.sub.2O was added and the precipitate was
collected and washed two times 50 ml Et.sub.2O and dried in vacuo.
The dried material was then stirred for 1 h. with a mixture of 2 ml
TFA, 50 .mu.l TIS and 50 .mu.l H.sub.2O and the crude pegylated
peptide was isolated by precipitation with 50 ml Et.sub.2O and
washed three times with 50 ml Et.sub.2O and then dried in vacuo.
The crude peptide was dissolved in 20 ml H.sub.2O and characterized
as follows: TABLE-US-00005 Analytical method Result HPLC A1 r.t.:
41.41 min. HPLC B6 r.t.: 30.06 min.
2.d Purification
[0797] The crude peptide in solution from 2.c was purified by
semipreparative HPLC in one run on a 25 mm.times.250 mm column
packed with 7 .mu.m C-18 silica. The column was eluted with a
gradient of 30 to 65% CH.sub.3CN in 0.1% TFA/H.sub.2O at 10 ml/min
at a temperature of 40.degree. C. for 47 min. The peptide
containing fractions corresponding to the major peak was collected,
diluted to 30 ml with approximately 3 volumes of H.sub.2O and
lyophilized. The final product obtained was characterized as
follows: TABLE-US-00006 Analytical method Result HPLC A1 r.t.:
41.12 min., HPLC B6 r.t.: 29.89 min., yield 47.9 mg. LC-MS A broad
peak at 3.45 min having a mass spectrum with a very large number of
mass peaks. Maldi TOF MS The mass spectrum displays a cluster of
peaks with an average mass of 8875 Da. This is in agreement with
the expected structure of the target compound.
Example 3
N.sup..epsilon.37-(3-(mPEGyl)propionyl)[Aib.sup.8,22,35,Lys.sup.37]GLP-1(7-
-37) wherein mPEGyl is Polydisperse and has a Molecular Weight of
Approximately 20 kDa
[0798] ##STR17##
[0799] This compound was prepared from 100 mg of crude protected
peptide from 2.b using procedures similar to those in example 2.c
and 2.b with the major exception that 400 mg 100 mg mPEG-20000-SPA
(mPEG-SPA m.w. 20.000 Lot PT-05C-11, Shearwater, Ala., USA) was
used for the pegylation.
[0800] The final product obtained was characterized as follows:
TABLE-US-00007 Analytical method Result HPLC A1 r.t.: 47.62 min.
HPLC B6 r.t.: 34.47 min. Maldi TOF MS The mass spectrum displays a
cluster of peaks with an average mass 25304 Da. This is in
agreement with the expected structure of the target compound.
Example 4
N.sup..epsilon.37-((2S)-2,6-di-(mPEGylcarbonylamino)hexanoyl)[Aib.sup.8,22-
,35,Lys.sup.37]GLP-1(7-37) wherein mPEGyl is Polydisperse and has A
Molecular Weight of Approximately 20 kDa
[0801] ##STR18##
[0802] This compound was prepared from 100 mg of crude protected
peptide from 2.b using procedures similar to those in example 2.c
and 2.b with the major exception that 800 mg mPEG2-40000-NHS ester
(mPEG2-NHS ester m.w. 40.000 Lot. PT-11C-06, Shearwater, Ala., USA)
was used for the pegylation.
[0803] The final product obtained was characterized as follows:
TABLE-US-00008 Analytical method Result HPLC B6 r.t.: 30.99 min.,
yield 3.07 mg. conjugate NMR .sup.1H-NMR showed that the 3.07 mg
conjugate isolated was contaminated with approximately 7 mg
hydrolysed pegylation reagent.
Example 5
N.sup..epsilon.26-(3-(mPEGyl)propionyl)[Aib.sup.8,Glu.sup.22,30,Lys.sup.33-
,Asn.sup.34,Gly.sup.35,36,Pro.sup.37]GLP-1(7-37)ylSerSerGly
AlaProProProSer amide wherein mPEGyl is Polydisperse and has a
Molecular Weight of Approximately 2 kDa
[0804] ##STR19##
[0805] The fully protected peptidyl resin
Boc-His(Boc)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)Ser(tBu)-Asp(OtBu)-Va-
l-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Leu-Glu(OtBu)-Glu(OtBu)-Gln-Ala-Ala-Lys(Dde)--
Glu(OtBu)-Phe-Ile-Glu(OtBu)-Trp(Boc)-Leu-Lys(Boc)-Asn(Trt)-Gly-Gly-Pro-Ser-
(tBu)Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Rink amide resin was
prepared using procedures similar to those used in example 1a. from
0.25 mmol Rink amide resin having a substitution capacity of 0.61
mmol/g. The yield was 1.4 g.
[0806] Analytical results from characterization of the resin as in
Example 1.b were: TABLE-US-00009 Analytical method Result HPLC A1
r.t.: 34.99 min., LC-MS r.t. 3.29 min., Mass for (M + 3H.sup.+)/3:
1432.8 Da, (calc.: 1433.2 Da)
[0807] The Dde protection was then removed from 350 mg of this
fully protected peptidyl resin using the procedures of example 1.c.
and the resulting resin was then pegylated using the procedure of
example 1.d using (mPEGyl)propionic acid 2,5-dioxo-pyrrolidin-1-yl
ester (Shearwater cat. no. 2M4M0D01, mPEG-SPA, MW 2,000) (0.5 g,
0.25 mmol, 4 eq.) and DIEA (43 .mu.l, 0.25 mmol, 4 eq.)
[0808] The pegylated peptide was then cleaved from the resin using
procedures similar to those of example i.e. and purified using a
procedures similar to those of example 1.f
[0809] The yield was 0.125 mg and the results from HPLC and LC-MS
analysis: TABLE-US-00010 Analytical method Result HPLC B6 r.t.:
37.45 min. estimated purity: 96.5% LG-MS r.t. 3.44 min., Average
mass for (M + 2H.sup.+)/2: 3074 Da
Example 6
N.sup..alpha.-[Aib.sup.8,22,35]GLP-1-(7-36)yl(N.sup..epsilon.-(3-(mPEGyl)p-
ropionyl)Lysinamide) wherein mPEGyl is Polydisperse and has a
Molecular Weight of Approximately 750 Da
[0810] ##STR20##
[0811] The fully protected peptidyl resin
Boc-His(Boc)-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)Ser(tBu)-Asp(OtBu)-Va-
l-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Leu-Glu(OtBu)-Aib-Gln(Trt)-Ala-Ala-Lys(Boc)-G-
lu(OtBu)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Lys(Boc)-Aib-Arg(Pmc)-Lys(ivDde)-PAL-
-PEG-PS resin was prepared using procedures similar to those used
in example 1a. from 0.25 mmol PAL-PEG-PS resin (Applied Biosystem
Cat. no. GEN 913398) having a substitution capacity of 0.38 mmol/g.
The yield was 1.935 g.
[0812] The ivDde protection was removed from the protected peptidyl
resin as follows. The resin (382 mg, 90 .mu.mol) was washed in NMP.
A freshly prepared solution of hydrazine hydrate 2% in NMP (20 ml)
was added and the reaction mixture was shaken for 12 min at room
temperature, and then filtered. The hydrazine treatment was
repeated twice. After this the resin was washed extensively with
NMP and coupled with
(N-(2-mPEGyl-ethyl)-4-(2,5-dioxo-pyrrolidin-1-yl)-4-oxo-butyramide
(.alpha.-Methoxy-.omega.-NHS ester PEG, Rapp Polymere GmbH,
Tubingen, FRG, cat no. 12 750-35) (0.27 g, 0.36 mmol, 4 eq.) using
the procedures of example 1.d.
[0813] The pegylated peptide was then cleaved and characterized
from the resin using procedures similar to those of example
i.e.
[0814] Results from HPLC and LC-MS analysis of the dry precipitate:
TABLE-US-00011 Analytical method Result HPLC A1 r.t.: 37.90 min.,
LC-MS r.t. 3.46 min., Average mass for (M+3H.sup.+)/3: 1410 Da
[0815] The crude peptide was finally purified using the procedures
similar to those of example 1.f. The yield was 26 mg product which
was characterized by HPLC and LC-MS TABLE-US-00012 Analytical
method Result HPLC A1 r.t.: 37.90 min., HPLC B1 r.t.: 39.46 min.
estimated purity: 98% LC-MS r.t. 3.48 min., Average mass for
(M+3H.sup.+)/3: 1410 Da
Example 7
N.sup..epsilon.-[Aib.sup.8,22,35]GLP-1(7-37)yl(S.sup..epsilon.-(1-mPEGylpr-
opyl-2,5-dioxo-pyrrolidin-3-yl)Cysteinamide wherein mPEGyl is
Polydisperse and has a Molecular Weight of Approximately 5000
Da
[0816] ##STR21##
[0817] The crude peptide
His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Aib-Gln-Ala-A-
la-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Aib-Arg-Gly-Cys(H) amide was
prepared using procedures similar to those used in example 1a., and
1.e from 0.25 mmol Rink amide resin having a substitution capacity
of 0.61 mmol/g. The yield was 121 mg
[0818] Results from HPLC and LC-MS analysis of the dry precipitate
of crude peptide: TABLE-US-00013 Analytical method Result HPLC A1
r.t.: 36.15 min., LC-MS r.t. 3.51 min., Mass for (M+3H.sup.+)/3:
1177.1.3 Da, (calc.: 1176 Da)
[0819] A fraction of the crude peptide (10 mg, 3 .mu.mol) was
dissolved in phosphate buffer (15 ml) and pH was adjusted to 6.5
and mPEG-Mal 5000 (Shearwater cat. no. 2D2M0H01, mPEG-MAL, MW
5,000) (28 mg, 6 .mu.mol) was added and the mixture was stirred for
30 min. The final product was isolated from this mixture using
procedures similar to those of example 1.f.
[0820] The yield was 2.2 mg and the results from HPLC and LC-MS
analysis: TABLE-US-00014 Analytical method Result HPLC A1 r.t.:
37.41 min., LC-MS r.t. 3.56 min., Average mass for (M+6H.sup.+)/6:
1058.1 Da,
Example 8
N.sup..alpha.-(3-(3H-imidazol-4-yl)-propionyl
[Aib.sup.22,35,Arg.sup.26,34]GLP-1-(8-37))yl(N.sup..epsilon.-(3-(mPEGyl)p-
ropionyl)Lysinamide) wherein mPEGyl is Polydisperse and has a
Molecular Weight of Approximately 2000 Da
[0821] ##STR22##
[0822] The fully protected peptidyl resin
ImPr(Adoc)-Ala-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Val-
-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Leu-Glu(OtBu)-Aib-Gln-Ala-Ala-Arg(Pmc)Glu(OtBu-
)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Arg(Pmc)-Aib-Arg(Pmc)-Gly-Lys(Boc)-Rink
amide resin using procedures similar to those used in example 1a in
1 mmol scale, with the exception that HATU was used as the coupling
agent throughout the synthesis. The resin used was a hydrophilic
Rink amide resin (HypoGel.RTM. 200 RAM) (Rapp Polymere cat. # SP200
110150 230) resin having a substitution capacity of 0.61
mmol/g.
[0823] Cleavage and purification was carried out as in the examples
1e and 1f. The yield was 210 mg and the results from HPLC and LC-MS
analysis:
[0824] Analytical method. Result: HPLC A1 r.t.: 36.51 min., LC-MS
r.t. 3.69 min., mass for (M+3H.sup.+)/3: 1194.4 Da, (calc.: 1193.4
Da)
[0825] The pegylation was performed as follows. 20 mg of the
unprotected peptide was dissolved in 600 .mu.l water and 100 mg of
the pegylation reagent (mPEGyl)propionic acid
2,5-dioxo-pyrrolidin-1-yl ester) (Shearwater cat. no. 2M4M0D01,
mPEG-SPA, MW 2,000) was added together with 9 .mu.l DIEA and
stirred for 24 h. The final product was isolated from this mixture
using procedures similar to those of example 1.f. The yield was 3.7
mg of the title compound and results from HPLC and MALDI analysis
were:
[0826] Analytical method. Result: HPLC 01_B4.sub.--2 r.t.: 10.96
min., MALDI-TOF Average mass for (M.sup.+) 5724 Da
Example 9
Preparation of
N.sup..epsilon.26-(3-(mPEGyl)propionyl)[Arg.sup.34]GLP-1-(7-37)
wherein mPEGyl is Polydisperse and has a Molecular Weight of
Approximately 2 kDa
[0827] ##STR23##
[0828] This compound was prepared by acylation in solution of
unprotected [Arg.sup.34]GLP-1-(7-37) which was obtained by
expression in yeast. [Arg34]GLP-1-(7-37) peptide (0.3 g, 30%
peptide content) was dissolved in water containing DIEA (101 .mu.l,
20 e.q.) and acylated with mPEG SPA 2000 (Shearwater Cat. no.
2M4M0D01, mPEG-SPA, MW 2,000) (89 mg, 1.5 e.q.) for 1 h at room
temperature. The final product was isolated from this mixture using
procedures similar to those of example 1.f. The yield of the title
compound was 61 mg and the results from HPLC and Maldi TOF MS
analysis analysis were: TABLE-US-00015 Analytical method Result
HPLC 02_B4_2 r.t.: 8.48 min., MALDI-TOF Average mass for (M.sup.+):
5587.3 Da
Example 10
(S)--N((S)-5-(N((S)-5-carbamoyl-5-(mPEGylpropionylamino)pentyl)carbamoyl)--
5-(mPEGylpropionylamino)pentyl)-5-(N.sup..alpha.7-(3-(4-imidazolyl)propion-
yl)[Aib.sup.22,35,Arg.sup.26,34]GLP-1-(8-37)yl)-2-(mPEGylpropionylamino)he-
xanoic amide wherein mPEGyl is polydisperse and has a molecular
weight of approximately 750 Da
[0829] ##STR24##
[0830] The fully protected peptidyl resin
Boc-Lys(Boc-Lys(Boc-Lys(ImPr(Adoc)-Ala-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu-
)-Ser(tBu)-Asp(OtBu)-Val-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Leu-Glu(OtBu)-Aib-Gln(-
Trt)-Ala-Ala-Arg(Pmc)-Glu(OtBu)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Arg(Pmc)-Aib--
Arg(Pmc))))Rink amide resin was synthesized using the procedures in
example 1.a. ImPr(Adoc)-OH was used for the introduction of the
N-terminal 3-(4-Imidazolyl)propinoyl group and Boc-Lys(Fmoc)OH was
used for introducing three side chain linked Lys residues in the
C-terminal of the sequence.
[0831] The corresponding unprotected peptide,
H-Lys(H-Lys(H-Lys(ImPr-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Le-
u-Glu-Aib-Gln-Ala-Ala-Arg-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Aib-Arg)))
amide, was cleaved from the resin using procedures similar to those
of example 1.e. and purified using procedures similar to those in
example 1.f.
[0832] The results from the characterization of this intermediate
peptide were: TABLE-US-00016 Analytical method Result HPLC A1 r.t.:
35.39 min., HPLC B1 r.t.: 36.50 min. LC-MS r.t. 3.85 min., Mass for
(M+3H+)/3: 1279.8 Da, (calc.: 1279.8 Da)
[0833] 3.84 mg this purified peptide dissolved it in 0.4 ml NMP and
pegylated by stirring at room temperature for 4 h with 8 mg with
(N-(2-mPEGyl-ethyl)-4-(2,5-dioxo-pyrrolidin-1-yl)-4-oxo-butyramide
(.alpha.-Methoxy-.omega.-NHS ester PEG, Rapp Polymere GmbH,
Tubingen, FRG, cat no. 12 750-35) and 7 .mu.l DIEA.
[0834] The title compound was finally purified using procedures
similar to those in example 1.f.
[0835] The results from the characterization were: TABLE-US-00017
Analytical method Result HPLC A1 r.t.: 41.62 min., HPLC B1 r.t.:
43.02 min., yield 6.5 mg. LC-MS A broad peak at 4.24 min having a
mass spectrum with a very large number of mass peaks. Maldi TOF MS
The mass spectrum displays a cluster of peaks with an average mass
of 6000 Da. This is in agreement with the expected structure of the
target compound.
Example 11
Method for Measuring Pulmonary Bioavailability.
[0836] The present protocol describes the methods and materials
used in the development of an anaesthetized rat model for pulmonal
delivery of aerosols. The aerosols are generated by use of a
nebulizer catheter with a well defined droplet/particle size (mean
mass aerodynamic diameter, MMAD). The nebulizer catheter is an
extruded multi-lumen catheter that provides fine-particle,
baffle-free, aerosols. It incorporates multiple (typically 4-6)
gas-lumens around one liquid lumen. Each lumen extends the length
of the catheter which tapers to a fine (.about.0.5 mm diameter)
nozzle with tiny orifices at the distal tip. The intimate contact
between the gas and liquid at the tip produces a fine aerosol
without baffling. The nebulizer catheter is guided through an
endotracheal tube and is placed just above the main bronchial
branch. The aerosol is deposited in pulses managed from a control
unit.
Equipment
[0837] The equipment for pulmonary delivery is obtained from
Trudell Medical International (London, Ontario, Canada).
Nebulizer Catheters
[0838] Nebulizer catheters (Aeroprobe.RTM.) are supplied from the
manufacturer in a number of different configurations and lengths.
These different designs will accommodate a variety of different
fluid and flow-rates, as well as provide aerosol particle-sizes
that may be as low as 5 .mu.m MMAD (mean mass aerodynamic
diameter). In the present experiments a catheter with the following
dimensions is used: Outer lumens gas flow of 1.4 L/min, inner lumen
liquid flow of 0.7 ml/min, and MMAD of about 7-8 .mu.m (PN
104504-050) with a length of 50 cm (1). Control Unit [0839]
LABNeb.RTM. Catheter control system (CCS). The Aeroprobe catheter
is connected to the control system according to (2).
[0840] Air with a pressure of 100 psi is used as supporting gas and
maximal fluid pressure, usually 98 psi. A 100 .mu.l syringe is used
as reservoir. The LABNeb CCS used a pulse time of 80 msec and a gas
delay of 20 msec. Thus, 2.3 ml air and 0.93 .mu.l test solution are
delivered in each pulse.
Animals
[0841] Sprague Dawley rats weighing between 250 and 350 g. The
animals are housed under standardised conditions with free access
to food (Altromine 1324) and drinking water. On the experimental
day the animals are used in their fed state.
Solution for Anaesthesia
[0842] Hypnorm.RTM. (fentanyl 0.2 mg/ml, fluansol 10 mg/ml) is
diluted with sterile water 1+1. Dormicum.RTM. (midazolam 5 mg/ml)
is diluted with sterile water 1+1. The two solutions are mixed
1+1.
Surgical Procedures and Intratracheal Administration
[0843] Anaesthesia is induced by injecting subcutaneously the
prepared Hyponorm/Dormicum solution 0.25 ml/100 g BW. An
endotracheal tube (PE 240, Becton Dickinson) is inserted and guided
to a position about 1/2 cm above the branch of the two main
bronchii. Any heat loss is minimised by wrapping a plastic shield
round the rat.
[0844] Before applying the test solution into the lungs, it is
secured that the syringe and catheter system is free of air
bubbles. Before applying the test solution endotracheally, it is
sprayed into a vial to test subsequently the amount of substance
administered by the catheter. Then, the catheter is guide through
the endotracheal tube leaving 1-2 mm of the catheter tip free of
the tube end and the test solution is aerosolised into the lungs of
the anaesthetised rat.
Sequence CWU 1
1
5 1 31 PRT homo sapiens 1 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 2 39 PRT heloderma suspectum
MOD_RES (39)..(39) Amidation of carboxy group 2 His Gly Glu Gly Thr
Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu 1 5 10 15 Glu Ala Val
Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30 Ser
Gly Ala Pro Pro Pro Ser 35 3 40 PRT artificial synthetic
MISC_FEATURE (1)..(1) Xaa at position 1 is L-histidine,
D-histidine, desamino-histidine, 2-amino-histidine,
beta-hydroxy-histidine, homohistidine, N-alpha-acetyl-histidine,
alpha-fluoromethyl- histidine, alpha-methyl-histidine,
3-pyridylalanine, 2-pyridylalanine, or 4-pyridylalanine.
MISC_FEATURE (2)..(2) Xaa at position 2 is Ala, D-Ala, Gly, Val,
Leu, Ile, Lys, Aib, (1-aminocyclopropyl) carboxylic acid,
(1-aminocyclobutyl) carboxylic acid,(1-aminocyclopentyl) carboxylic
acid, (1-aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl)
carboxylic acid or (1-aminocyclooctyl) carboxylic acid.
MISC_FEATURE (10)..(10) Xaa at position 10 is Val or Leu.
MISC_FEATURE (12)..(12) Xaa at position 12 is Ser, Lys or Arg.
MISC_FEATURE (13)..(13) Xaa at position 13 is Tyr or Gln.
MISC_FEATURE (14)..(14) Xaa at position 14 is Leu or Met.
MISC_FEATURE (16)..(16) Xaa at position 16 is Gly, Glu or Aib.
MISC_FEATURE (17)..(17) Xaa at position 17 is Gln, Glu, Lys or Arg.
MISC_FEATURE (19)..(19) Xaa at position 19 is Ala or Val.
MISC_FEATURE (20)..(20) Xaa at position 20 is Lys, Glu or Arg.
MISC_FEATURE (21)..(21) Xaa at position 21 is Glu or Leu.
MISC_FEATURE (24)..(24) Xaa at position 24 is Ala, Glu or Arg.
MISC_FEATURE (27)..(27) Xaa at position 27 is Val or Lys.
MISC_FEATURE (28)..(28) Xaa at position 28 is Lys, Glu, Asn or Arg.
MISC_FEATURE (29)..(29) Xaa at position 29 is Gly or Aib.
MISC_FEATURE (30)..(30) Xaa at position 30 is Arg, Gly or Lys.
MISC_FEATURE (31)..(31) Xaa at position 31 is Gly, Ala, Glu, Pro,
Lys, or is absent MISC_FEATURE (32)..(32) Xaa at position 32 is
Lys, Ser, or is absent. MISC_FEATURE (33)..(33) Xaa at position 33
is Ser, Lys, or is absent. MISC_FEATURE (34)..(34) Xaa at position
34 is Gly, or is absent. MISC_FEATURE (35)..(35) Xaa at position 35
is Ala, or is absent. MISC_FEATURE (36)..(36) Xaa at position 36 is
Pro, or is absent. MISC_FEATURE (37)..(37) Xaa at position 37 is
Pro, or is absent. MISC_FEATURE (38)..(38) Xaa at position 38 is
Pro, or is absent. MISC_FEATURE (39)..(39) Xaa at position 39 is
Ser, or is absent. MISC_FEATURE (40)..(40) Xaa at position 40 is
absent. 3 Xaa Xaa Glu Gly Thr Phe Thr Ser Asp Xaa Ser Xaa Xaa Xaa
Glu Xaa 1 5 10 15 Xaa Ala Xaa Xaa Xaa Phe Ile Xaa Trp Leu Xaa Xaa
Xaa Xaa Xaa Xaa 20 25 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 35 40 4 32
PRT artificial synthetic MISC_FEATURE (1)..(1) Xaa at position 1 is
L-histidine, D-histidine, desamino-histidine, 2-amino-histidine,
beta-hydroxy-histidine, homohistidine, N-alpha-acetyl-histidine,
alpha-fluoromethyl- histidine, alpha-methyl-histidine,
3-pyridylalanine, 2-pyridylalanine, or 4-pyridylalanine.
MISC_FEATURE (2)..(2) Xaa at position 2 is Ala, D-Ala, Gly, Val,
Leu, Ile, Lys, Aib, (1-aminocyclopropyl) carboxylic acid,
(1-aminocyclobutyl) carboxylic acid, (1-aminocyclopentyl)
carboxylic acid, (1-aminocyclohexyl) carboxylic acid,
(1-aminocycloheptyl) carboxylic acid or (1-aminocyclooctyl)
carboxylic acid. MISC_FEATURE (12)..(12) Xaa at position 12 is Ser,
Lys or Arg. MISC_FEATURE (16)..(16) Xaa at position 16 is Gly, Glu
or Aib. MISC_FEATURE (17)..(17) Xaa at position 17 is Gln, Glu, Lys
or Arg. MISC_FEATURE (20)..(20) Xaa at position 20 is Lys, Glu or
Arg. MISC_FEATURE (24)..(24) Xaa at position 24 is Ala, Glu or Arg.
MISC_FEATURE (28)..(28) Xaa at position 28 is Lys, Glu or Arg.
MISC_FEATURE (29)..(29) Xaa at position 29 is Gly or Aib.
MISC_FEATURE (30)..(30) Xaa at position 30 is Arg or Lys.
MISC_FEATURE (31)..(31) Xaa at position 31 is Gly, Ala, Glu or Lys.
MISC_FEATURE (32)..(32) Xaa at position 32 is Lys, or is absent. 4
Xaa Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Xaa Tyr Leu Glu Xaa 1 5
10 15 Xaa Ala Ala Xaa Glu Phe Ile Xaa Trp Leu Val Xaa Xaa Xaa Xaa
Xaa 20 25 30 5 44 PRT artificial sequence MOD_RES (44)..(44)
Amidation of carboxyl group 5 His Gly Glu Gly Thr Phe Thr Ser Asp
Leu Ser Lys Gln Met Glu Glu 1 5 10 15 Glu Ala Val Arg Leu Phe Ile
Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30 Ser Gly Ala Pro Pro
Ser Lys Lys Lys Lys Lys Lys 35 40
* * * * *