U.S. patent application number 10/372485 was filed with the patent office on 2003-11-27 for inhibition of beta cell degeneration.
Invention is credited to Carr, Richard David, Godtfredsen, Carsten Foged, Knudsen, Liselotte Bjerre, Petersen, Jacob Sten.
Application Number | 20030220251 10/372485 |
Document ID | / |
Family ID | 27512869 |
Filed Date | 2003-11-27 |
United States Patent
Application |
20030220251 |
Kind Code |
A1 |
Knudsen, Liselotte Bjerre ;
et al. |
November 27, 2003 |
Inhibition of beta cell degeneration
Abstract
This invention relates to a method for modulating, inhibiting or
decreasing or preventing beta cell degeneration, loss of beta cell
function, beta cell dysfunction, and/or death of beta cells, such
as necrosis or apoptosis of beta cells in a subject comprising
administering a GLP-1 agonist to said subject.
Inventors: |
Knudsen, Liselotte Bjerre;
(Valby, DK) ; Godtfredsen, Carsten Foged; (Herlev,
DK) ; Petersen, Jacob Sten; (Copenhagen O, DK)
; Carr, Richard David; (Vaerlose, DK) |
Correspondence
Address: |
Reza Green, Esq.
Novo Nordisk Pharmaceuticals, Inc.
100 College Road West
Princeton
NJ
08540
US
|
Family ID: |
27512869 |
Appl. No.: |
10/372485 |
Filed: |
February 24, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10372485 |
Feb 24, 2003 |
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09709856 |
Nov 10, 2000 |
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6569832 |
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60166800 |
Nov 22, 1999 |
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60185845 |
Feb 29, 2000 |
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Current U.S.
Class: |
514/11.7 ;
514/18.9 |
Current CPC
Class: |
A61P 5/50 20180101; A61K
38/26 20130101 |
Class at
Publication: |
514/12 |
International
Class: |
A61K 038/26 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 12, 1999 |
DK |
PA 1999 01628 |
Feb 22, 2000 |
DK |
PA 2000 00270 |
Claims
1. Use of a GLP-1 agonist for the preparation of a medicament for
treatment of beta cell degeneration.
2. The use according to claim 1 wherein the beta cell degeneration
is apoptosis of .beta.-cells.
3. The use according to claim 1 or 2 wherein the GLP-1 agonist is
selected from a GLP-1 analogue, a GLP-1 derivative wherein at least
one amino acid residue of the parent peptide has a lipophilic
substituent attached, exendin or an analogue or derivative thereof,
or a non-peptide, which binds to a GLP-1 receptor with an affinity
constant, K.sub.D, below 1 .mu.M.
4. The use according to claim 1, 2 or 3 wherein the GLP-1
derivative is Arg.sup.34,
Lys.sup.26(N-.epsilon.-(.gamma.-Glu(N-.alpha.-hexadecanoyl)))-
-GLP-1(7-37).
5. A method for treatment of beta cell degeneration in a subject
comprising administering a GLP-1 agonist to said subject.
6. The method according to claim 5 wherein the beta cell
degeneration is apoptosis of .beta.-cells.
7. The method according to claim 5 or 6 wherein the GLP-1 agonist
is selected from a GLP-1 analogue, a GLP-1 derivative wherein at
least one amino acid residue of the parent peptide has a lipophilic
substituent attached, exendin or an analogue or derivative thereof,
or a non-peptide, which binds to a GLP-1 receptor with an affinity
constant, K.sub.D, below 1 .mu.M.
8. The method according to claim 5, 6 or 7 wherein the GLP-1
derivative is Arg.sup.34,
Lys.sup.26(N-.epsilon.-(.gamma.-Glu(N-.alpha.-hexadecanoyl)))-
-GLP-1(7-37).
Description
[0001] The present invention relates to a method for modulating,
inhibiting or decreasing or preventing beta cell degeneration, loss
of beta cell function, beta cell dysfunction, and/or death of beta
cells, such as necrosis or apoptosis of beta cells in a subject
comprising administering a GLP-1 agonist to said subject.
BACKGROUND
[0002] What most textbooks of pathology describe as cell death is
coagulative necrosis. This is an abnormal morphological appearance,
detected in tissue examined under the microscope. The changes,
which affect aggregates of adjacent cells or functionally related
cohorts of cells, are seen in a variety of contexts produced by
accident, injury, or disease. Among the environmental perturbations
that may cause cell necrosis are oxygen deprivation (anoxia),
hyper-thermia, immunological attack, and exposure to various toxins
that inhibit crucial intracellular metabolic processes. Coagulative
necrosis is the classical form of cell change seen when tissues
autolyze (digest themselves) in vitro.
[0003] Apoptosis is an active process of cellular self-destruction
that is regulated by extrinsic and intrinsic signals occurring
during normal development. It is well documented that apoptosis
plays a key role in regulation of pancreatic endocrine beta cells.
There is increasing evidence that in adult mammalians the beta-cell
mass is submitted to dynamic changes to adapt insulin production
for maintaining euglycemia in particular conditions, such as
pregnancy and obesity (J. Dev. Physiol. 5: 373, 1983 and
Endocrinology 130: 1459, 1992). The control of beta cell mass
depends on a subtle balance between cell proliferation, growth and
cell death (apoptosis). A disruption of this balance may lead to
impairment of glucose homeostasis. For example, it is noteworthy
that glucose intolerance develops with aging when beta cell
replication rates are reduced (Diabetes 32: 14, 1983) and human
autopsy studies repeatedly showed a 40-60% reduction of beta cell
mass in patients with non-insulin-dependent-diabetes mellitus
compared with nondiabetic subjects (Am. J. Med. 70: 105, 1981 and
Diabetes Res. 9: 151, 1988). It is generally agreed that insulin
resistance is an invariable accompaniment of obesity but that
normoglycemia is maintained by compensatory hyperinsulinemia until
the beta cells become unable to meet the increased demand for
insulin, at which point Type 2 Diabetes begins.
[0004] Apoptosis is also associated with diseases states such as
cancer, immunological disorders, like multiple sclerosis, and AIDS,
and neurodegenerative disorders (Science 267: 1449, 1995), like
Alzheimers disease, stroke, and Parkinson's disease.
DESCRIPTION OF THE INVENTION
[0005] Accordingly, the present invention relates to use of a GLP-1
agonist for the preparation of a medicament for treatment of beta
cell degeneration, such as necrosis or apoptosis of
.beta.-cells.
[0006] Furthermore, the present invention relates to use of a GLP-1
agonist for the preparation of a medicament for modulation of beta
cell degeneration, such as necrosis or apoptosis of
.beta.-cells.
[0007] Furthermore, the present invention relates to use of a GLP-1
agonist for the preparation of a medicament for inhibition of beta
cell degeneration, such as necrosis or apoptosis of
.beta.-cells.
[0008] Furthermore, the present invention relates to use of a GLP-1
agonist for the preparation of a medicament for decreasing beta
cell degeneration, such as necrosis or apoptosis of
.beta.-cells.
[0009] Furthermore, the present invention relates to use of a GLP-1
agonist for the preparation of a medicament for reduction of beta
cell degeneration, such as necrosis or apoptosis of
.beta.-cells.
[0010] Furthermore, the present invention relates to use of a GLP-1
agonist for the preparation of a medicament for arresting beta cell
degeneration, such as necrosis or apoptosis of .beta.-cells.
[0011] Furthermore, the present invention relates to use of a GLP-1
agonist preventing beta cell degeneration for the preparation of a
medicament for prevention of beta cell degeneration, such as
necrosis or apoptosis of .beta.-cells.
[0012] Moreover, the invention relates to a method for treatment of
beta cell degeneration, such as necrosis or apoptosis of
.beta.-cells, in a subject comprising administering a GLP-1 agonist
to said subject.
[0013] Furthermore, the invention relates to a method for
modulation of beta cell degeneration, such as necrosis or apoptosis
of .beta.-cells, in a subject comprising administering a GLP-1
agonist to said subject.
[0014] Furthermore, the invention relates to a method for
inhibition of beta cell degeneration, such as necrosis or apoptosis
of .beta.-cells, in a subject comprising administering a GLP-1
agonist to said subject.
[0015] Furthermore, the invention relates to a method for
decreasing beta cell degeneration, such as necrosis or apoptosis of
.beta.-cells, in a subject comprising administering a GLP-1 agonist
to said subject.
[0016] Furthermore, the invention relates to a method for reduction
of beta cell degeneration, such as necrosis or apoptosis of
.beta.-cells, in a subject comprising administering a GLP-1 agonist
to said subject.
[0017] Furthermore, the invention relates to a method for arresting
beta cell degeneration, such as necrosis or apoptosis of
.beta.-cells, in a subject comprising administering a GLP-1 agonist
to said subject.
[0018] Furthermore, the invention relates to a method for
prevention of beta cell degeneration, such as necrosis or apoptosis
of .beta.-cells, in a subject comprising administering a GLP-1
agonist to said subject.
[0019] In one embodiment of the invention beta cell degeneration is
necrosis of beta cells.
[0020] In another embodiment of the invention beta cell
degeneration is apoptosis of beta cells. In a further embodiment
said apoptosis is induced by a cytokine. The cytokine may be any
cytokine or mixtures thereof, such as interleukin 1 (IL-1), IL-2,
IL-3, IL-5, IL-6, IL-7, IL-8, IL-9, IL-12, IL-14, IL-17,
interferon-.gamma., tumor necrosis factor-.alpha., TNF-.beta.,
granulocyte macrophage colony stimulating factor, monocyte
chemoattractant protein-1, or mixtures thereof.
[0021] The subject is preferably a mammal, more preferably a
human.
[0022] The use according any of the above uses in a regimen which
additionally comprises treatment with human growth hormone, a
growth hormone releasing agent or a growth factor such as prolactin
or placental lactogen; the use of human growth hormone, a growth
hormone releasing agent or a growth factor such as prolactin or
placental lactogen for the preparation of a medicament for
inhibiting the beta cell degeneration, such as necrosis or
apoptosis of .beta.-cells in a subject; the use of human growth
hormone, a growth hormone releasing agent or a growth factor such
as prolactin or placental lactogen for the preparation of a
medicament for treatment of beta cell degeneration, such as
necrosis or apoptosis of .beta.-cells in a subject.
[0023] In one embodiment of the invention the GLP-1 agonist is
GLP-1(7-37) or GLP-1(7-36) amide.
[0024] In a further embodiment of the invention the GLP-1 agonist
is a GLP-1 analogue.
[0025] In a further embodiment of the invention the GLP-1 analogue
is selected from the Thr.sup.8, Met.sup.8, Gly.sup.8 and Val.sup.8
analogues of GLP-1(7-37) and GLP-1(7-36) amide, more preferred the
Gly.sup.8 and Val.sup.8 analogues of GLP-1(7-37) and GLP-1(7-36)
amide, most preferred the Val.sup.8 analogues of GLP-1(7-37) and
GLP-1(7-36) amide.
[0026] In a further embodiment of the invention the GLP-1 analogue
has the formula II:
1 7 8 9 10 11 12 13 14 15 16 17 His-Xaa-Xaa-Gly-Xaa-Phe--
Thr-Xaa-Asp-Xaa-Xaa- 18 19 20 21 22 23 24 25 26 27 28
Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Phe- 29 30 31 32 33 34 35
36 37 38 Ile-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa 39 40 41 42 43 44
45 Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa (II)
[0027] wherein
[0028] Xaa at position 8 is Ala, Gly, Ser, Thr, Leu, Ile, Val, Glu,
Asp, Met, or Lys,
[0029] Xaa at position 9 is Glu, Asp, or Lys,
[0030] Xaa at position 11 is Thr, Ala, Gly, Ser, Leu, Ile, Val,
Glu, Asp, or Lys,
[0031] Xaa at position 14 is Ser, Ala, Gly, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0032] Xaa at position 16 is Val, Ala, Gly, Ser, Thr, Leu, Ile,
Tyr, Glu, Asp, or Lys,
[0033] Xaa at position 17 is Ser, Ala, Gly, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0034] Xaa at position 18 is Ser, Ala, Gly, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0035] Xaa at position 19 is Tyr, Phe, Trp, Glu, Asp, or Lys,
[0036] Xaa at position 20 is Leu, Ala, Gly, Ser, Thr, Leu, Ile,
Val, Glu, Asp, or Lys,
[0037] Xaa at position 21 is Glu, Asp, or Lys,
[0038] Xaa at position 22 is Gly, Ala, Ser, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0039] Xaa at position 23 is Gln, Asn, Arg, Glu, Asp, or Lys,
[0040] Xaa at position 24 is Ala, Gly, Ser, Thr, Leu, Ile, Val,
Arg, Glu, Asp, or Lys,
[0041] Xaa at position 25 is Ala, Gly, Ser, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0042] Xaa at position 26 is Lys, Arg, Gln, Glu, Asp, or His,
[0043] Xaa at position 27 is Glu, Asp, or Lys,
[0044] Xaa at position 30 is Ala, Gly, Ser, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0045] Xaa at position 31 is Trp, Phe, Tyr, Glu, Asp, or Lys,
[0046] Xaa at position 32 is Leu, Gly, Ala, Ser, Thr, Ile, Val,
Glu, Asp, or Lys,
[0047] Xaa at position 33 is Val, Gly, Ala, Ser, Thr, Leu, Ile,
Glu, Asp, or Lys,
[0048] Xaa at position 34 is Lys, Arg, Glu, Asp, or His,
[0049] Xaa at position 35 is Gly, Ala, Ser, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0050] Xaa at position 36 is Arg, Lys, Glu, Asp, or His,
[0051] Xaa at position 37 is Gly, Ala, Ser, Thr, Leu, Ile, Val,
Glu, Asp, or Lys, or is deleted,
[0052] Xaa at position 38 is Arg, Lys, Glu, Asp, or His, or is
deleted,
[0053] Xaa at position 39 is Arg, Lys, Glu, Asp, or His, or is
deleted,
[0054] Xaa at position 40 is Asp, Glu, or Lys, or is deleted,
[0055] Xaa at position 41 is Phe, Trp, Tyr, Glu, Asp, or Lys, or is
deleted,
[0056] Xaa at position 42 is Pro, Lys, Glu, or Asp, or is
deleted,
[0057] Xaa at position 43 is Glu, Asp, or Lys, or is deleted,
[0058] Xaa at position 44 is Glu, Asp, or Lys, or is deleted,
and
[0059] Xaa at position 45 is Val, Glu, Asp, or Lys, or is deleted,
or
[0060] (a) a C-1-6-ester thereof, (b) amide, C-1-6-alkylamide, or
C-1-6-dialkylamide thereof and/or (c) a pharmaceutically acceptable
salt thereof,
[0061] provided that
[0062] (i) when the amino acid at position 37, 38, 39, 40, 41, 42,
43 or 44 is deleted, then each amino acid downstream of the amino
acid is also deleted.
[0063] In a further embodiment of the GLP-1 analogue of formula II,
the amino acids at positions 37-45 are absent.
[0064] In another embodiment of the GLP-1 analogue of formula II,
the amino acids at positions 38-45 are absent.
[0065] In another embodiment of the GLP-1 analogue of formula II,
the amino acids at positions 39-45 are absent.
[0066] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 8 is Ala, Gly, Ser. Thr, Met, or Val.
[0067] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 8 is Gly, Thr, Met, or Val.
[0068] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 8 is Val.
[0069] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 9 is Glu.
[0070] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 11 is T.
[0071] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 14 is Ser.
[0072] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 16 is Val.
[0073] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 17 is Ser.
[0074] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 18 is Ser, Lys, Glu, or Asp.
[0075] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 19 is Tyr, Lys, Glu, or Asp.
[0076] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 20 is Leu, Lys, Glu, or Asp.
[0077] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 21 is Glu, Lys, or Asp.
[0078] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 22 is Gly, Glu, Asp, or Lys.
[0079] In another embodiment of the GLP-1 analogue of formula IT,
Xaa at position 23 is Gln, Glu, Asp, or Lys.
[0080] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 24 is Ala, Glu, Asp, or Lys.
[0081] In another embodiment of the GLP-1 analogue of formula IT,
Xaa at position 25 is Ala, Glu, Asp, or Lys.
[0082] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 26 is Lys, Glu, Asp, or Arg.
[0083] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 27 is Glu, Asp, or Lys.
[0084] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 30 is Ala, Glu, Asp, or Lys.
[0085] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 31 is Trp, Glu, Asp, or Lys.
[0086] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 32 is Leu, Glu, Asp, or Lys.
[0087] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 33 is Val, Glu, Asp, or Lys.
[0088] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 34 is Lys, Arg, Glu, or Asp.
[0089] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 35 is Gly, Glu, Asp, or Lys.
[0090] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 36 is Arg, Lys, Glu, or Asp.
[0091] In another embodiment of the GLP-1 analogue of formula IT,
Xaa at position 37 is Gly, Glu, Asp, or Lys.
[0092] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 38 is Arg, or Lys, or is deleted.
[0093] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 39 is deleted.
[0094] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 40 is deleted.
[0095] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 41 is deleted.
[0096] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 42 is deleted.
[0097] In another embodiment of the GLP-1 analogue of formula IT,
Xaa at position 43 is deleted.
[0098] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 44 is deleted.
[0099] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 45 is deleted.
[0100] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 26 is Arg, each of Xaa at positions 37-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-36).
[0101] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 26 is Arg, each of Xaa at positions 38-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-37).
[0102] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 26 is Arg, each of Xaa at positions 39-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-38).
[0103] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 34 is Arg, each of Xaa at positions 37-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-36).
[0104] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 34 is Arg, each of Xaa at positions 38-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-37).
[0105] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 34 is Arg, each of Xaa at positions 3945 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-38).
[0106] In another embodiment of the GLP-1 analogue of formula II,
Xaa at positions 26 and 34 is Arg, Xaa at position 36 is Lys, each
of Xaa at positions 37-45 is deleted, and each of the other Xaa is
the amino acid in native GLP-1(7-36).
[0107] In another embodiment of the GLP-1 analogue of formula II,
Xaa at positions 26 and 34 is Arg, Xaa at position 36 is Lys, each
of Xaa at positions 38-45 is deleted, and each of the other Xaa is
the amino acid in native GLP-1(7-37).
[0108] In another embodiment of the GLP-1 analogue of formula II,
Xaa at positions 26 and 34 is Arg, Xaa at position 36 is Lys, each
of Xaa at positions 39-45 is deleted, and each of the other Xaa is
the amino acid in native GLP-1(7-38).
[0109] In another embodiment of the GLP-1 analogue of formula II,
Xaa at positions 26 and 34 is Arg, Xaa at position 38 is Lys, each
of Xaa at positions 39-45 is deleted, and each of the other Xaa is
the amino acid in native GLP-1(7-38).
[0110] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 8 is Thr, Ser, Gly, or Val, Xaa at position 37 is
Glu, Xaa at position 36 is Lys, each of Xaa at positions 38-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-37).
[0111] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 8 is Thr, Ser, Gly, or Val, Xaa at position 37 is
Glu, Xaa at position 36 is Lys, each of Xaa at positions 39-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-38).
[0112] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 8 is Thr, Ser, Gly or Val, Xaa at position 37 is
Glu, Xaa at position 38 is Lys, each of Xaa at positions 39-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-38).
[0113] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 18, 23 or 27 is Lys, and Xaa at positions 26 and 34
is Arg, each of Xaa at positions 37-45 is deleted, and each of the
other Xaa is the amino acid in native GLP-1(7-36).
[0114] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 18, 23 or 27 is Lys, and Xaa at positions 26 and 34
is Arg, each of Xaa at positions 38-45 is deleted, and each of the
other Xaa is the amino acid in native GLP-1(7-37).
[0115] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 18, 23 or 27 is Lys, and Xaa at positions 26 and 34
is Arg, each of Xaa at positions 39-45 is deleted, and each of the
other Xaa is the amino acid in native GLP-1(7-38).
[0116] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 8 is Thr, Ser, Gly, or Val, Xaa at position 18, 23
or 27 is Lys, and Xaa at position 26 and 34 is Arg, each of Xaa at
positions 37-45 is deleted, and each of the other Xaa is the amino
acid in native GLP-1(7-36).
[0117] In another embodiment of the GLP-1 analogue of formula II,
Xaa at position 8 is Thr, Ser, Gly, or Val, Xaa at position 18, 23
or 27 is Lys, and Xaa at position 26 and 34 is Arg, each of Xaa at
positions 38-45 is deleted, and each of the other Xaa is the amino
acid in native GLP-1(7-37).
[0118] In another embodiment of the GLP-1 analogue of formula IT,
Xaa at position 8 is Thr, Ser, Gly, or Val, Xaa at position 18, 23
or 27 is Lys, and Xaa at position 26 and 34 is Arg, each of Xaa at
positions 39-45 is deleted, and each of the other Xaa is the amino
acid in native GLP-1(7-38).
[0119] Such GLP-1 analogues includes, but is not limited to,
Arg.sup.26-GLP-1(7-37); Arg.sup.34-GLP-1(7-37);
Lys.sup.36-GLP-1(7-37); Arg.sup.26,34Lys.sup.36-GLP-1(7-37);
Arg.sup.26,34Lys 38GLP-1(7-38);
Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Arg.sup.26,34Lys.sup.40-GLP-1(7-40);
Arg.sup.26Lys.sup.36-GLP-1(7-37); Arg.sup.34Lys.sup.36-GLP-1(7-37);
Arg.sup.26Lys.sup.39-GLP-1(7-39); Arg.sup.34Lys.sup.40-GLP-1(7-40);
Arg.sup.26,34Lys.sup.36,39-GLP-1(7-39);
Arg.sup.26,34Lys.sup.36,40-GLP-1(- 7-40);
Gly.sup.8Arg26-GLP-1(7-37); Gly.sup.8Arg.sup.34-GLP-1(7-37);
Val.sup.8-GLP-1(7-37); Thr.sup.8-GLP-1(7-37);
Gly.sup.8-GLP-1(7-37); Met.sup.8-GLP-1(7-37);
Gly.sup.8Lys.sup.36-GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys.s- up.39-GLP-1(7-39);
Gly.sup.8Arg.sup.26,34Lys.sup.40-GLP-1(7-40);
Gly.sup.8Arg.sup.26Lys.sup.36-GLP-1(7-37);
Gly.sup.8Arg.sup.34Lys.sup.36-- GLP-1(7-37);
Gly.sup.8Arg.sup.26Lys.sup.39-GLP-1(7-39);
Gly.sup.8Arg.sup.34Lys.sup.40-GLP-1(7-40);
Gly.sup.8Arg.sup.26,34Lys.sup.- 36,39GLP-1(7-39);
Gly.sup.8Arg.sup.23,34Lys.sup.36,40-GLP-1(7-40);
Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Arg.sup.26,34Lys.sup.39GLP-1(7-39);
Arg.sup.26,34Lys.sup.40GLP-1(7-40);
Arg.sup.26,34Lys.sup.41GLP-1(7-41);
Arg.sup.26,34Lys.sup.42GLP-1(7-42);
Arg.sup.26,34Lys.sup.43GLP-1(7-43);
Arg.sup.26,34Lys.sup.44GLP-1(7-44);
Arg.sup.26,34Lys.sup.45GLP-1(7-45);
Arg.sup.26,34Lys.sup.38GLP-1(1-38);
Arg.sup.26,34Lys.sup.39GLP-1(1-39);
Arg.sup.26,34Lys.sup.40GLP-1(1-40);
Arg.sup.26,34Lys.sup.41GLP-1(1-41);
Arg.sup.26,34Lys.sup.42GLP-1(1-42);
Arg.sup.26,34Lys.sup.43GLP-1(1-43);
Arg.sup.26,34Lys.sup.44GLP-1(1-44);
Arg.sup.26,34Lys.sup.45GLP-1(1-45);
Arg.sup.26,34Lys.sup.38GLP-1(2-38);
Arg.sup.26,34Lys.sup.39GLP-1(2-39);
Arg.sup.26,34Lys.sup.40GLP-1(2-40);
Arg.sup.26,34Lys.sup.41GLP-1(2-41);
Arg.sup.26,34Lys.sup.42GLP-1(2-42);
Arg.sup.26,34Lys.sup.43GLP-1(2-43);
Arg.sup.26,34Lys.sup.44GLP-1(2-44);
Arg.sup.26,34Lys.sup.41GLP-1(2-45);
Arg.sup.26,34Lys.sup.38GLP-1(3-38);
Arg.sup.26,34Lys.sup.39GLP-1(3-39);
Arg.sup.26,34Lys.sup.40GLP-1(3-40);
Arg.sup.26,34Lys.sup.41GLP-1(3-41);
Arg.sup.26,34Lys.sup.42GLP-1(3-42);
Arg.sup.26,34Lys.sup.43GLP-1(3-43);
Arg.sup.26,34Lys.sup.44GLP-1(3-44);
Arg.sup.26,34Lys.sup.45GLP-1(3-45);
Arg.sup.26,34Lys.sup.38GLP-1(4-38);
Arg.sup.26,34Lys.sup.39GLP-1(4-39);
Arg.sup.26,34Lys.sup.40GLP-(4-40);
Arg.sup.26,34Lys.sup.41GLP-1(4-41);
Arg.sup.26,34Lys.sup.42GLP-1(4-42);
Arg.sup.26,34Lys.sup.43GLP-1(4-43);
Arg.sup.26,34Lys.sup.44GLP-1(4-44);
Arg.sup.26,34Lys.sup.45GLP-1(4-45);
Arg.sup.26,34Lys.sup.38GLP-1(5-38);
Arg.sup.26,34Lys.sup.34GLP-1(5-39);
Arg.sup.26,34Lys.sup.40GLP-1(5-40);
Arg.sup.26,34Lys.sup.41GLP-1(5-41);
Arg.sup.26,34Lys.sup.42GLP-1(5-42);
Arg.sup.26,34Lys.sup.43GLP-1(5-43);
Arg.sup.26,34Lys.sup.44GLP-1(5-44);
Arg.sup.26,34Lys.sup.45GLP-1(2-45);
Arg.sup.26,34Lys.sup.38GLP-1(6-38);
Arg.sup.26,34Lys.sup.39GLP-1(6-39);
Arg.sup.21,34Lys.sup.40GLP-1(6-40);
Arg.sup.26,34Lys.sup.41GLP-1(6-41);
Arg.sup.26,34Lys.sup.42GLP-1(6-42);
Arg.sup.26,34Lys.sup.43GLP-1(6-43);
Arg.sup.26,34Lys.sup.44GLP-1(6-44);
Arg.sup.26,34Lys.sup.45GLP-1(6-45);
Arg.sup.26Lys.sup.38GLP-1(1-38); Arg.sup.34Lys.sup.38GLP-1(1-38);
Arg.sup.26,34Lys.sup.36,38GLP-1(1-38);
Arg.sup.26Lys.sup.38GLP-1(7-38); Arg.sup.34Lys.sup.38GLP-1(7-38);
Arg.sup.26,34Lys.sup.36,38GLP-1(7-38);
Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Arg.sup.26Lys.sup.39GLP-1(1-39); Arg.sup.34Lys.sup.39GLP-1(1-39);
Arg.sup.26,34Lys.sup.36,39GLP-1(1-39);
Arg.sup.26Lys.sup.39GLP-1(7-39); Arg.sup.34Lys.sup.39GLP-1(7-39)
and Arg.sup.26,39Lys.sup.36,39GLP-1(7-39). Each one of these
specific GLP-1 analogues constitutes an alternative embodiment of
the invention.
[0120] In a still further embodiment of the invention the GLP-1
agonist is a GLP-1 derivative.
[0121] In a further embodiment of the invention the GLP-1
derivative has one or more lipophilic substituents attached to the
parent peptide. The lipophilic substituents make the profile of
action of the parent GLP-1 peptide more protracted, make the parent
GLP-1 peptide more metabolically and physically stable, and/or
increase the water solubility of the parent GLP-1 peptide.
[0122] The lipophilic substituent is characterised by having a
solubility in water at 20.degree. C. in the range from about 0.1
mg/100 ml water to about 250 mg/100 ml water, preferable in the
range from about 0.3 mg/100 ml water to about 75 mg/100 ml water.
For instance, octanoic acid (C8) has a solubility in water at
20.degree. C. of 68 mg/100 ml, decanoic acid (C10) has a solubility
in water at 20.degree. C. of 15 mg/100 ml, and octadecanoic acid
(C18) has a solubility in water at 20.degree. C. of 0.3 mg/100
ml.
[0123] In a further embodiment of the invention the GLP-1
derivatives preferably have three lipophilic substituents, more
preferably two lipophilic substituents, and most preferably one
lipophilic substituent.
[0124] Each lipophilic substituent(s) preferably has 4-40 carbon
atoms, more preferably 8-30 carbon atoms, even more preferably 8-25
carbon atoms, even more preferably 12-25 carbon atoms, and most
preferably 14-18 carbon atoms.
[0125] The lipophilic substituent(s) contain a functional group
which can be attached to one of the following functional groups of
an amino acid of the parent GLP-1 peptide:
[0126] (a) the amino group attached to the alpha-carbon of the
N-terminal amino acid,
[0127] (b) the carboxy group attached to the alpha-carbon of the
C-terminal amino acid,
[0128] (c) the epsilon-amino group of any Lys residue,
[0129] (d) the carboxy group of the R group of any Asp and Glu
residue,
[0130] (e) the hydroxy group of the R group of any Tyr, Ser and Thr
residue,
[0131] (f) the amino group of the R group of any Trp, Asn, Gln,
Arg, and His residue, or
[0132] (g) the thiol group of the R group of any Cys residue.
[0133] In an embodiment, a lipophilic substituent is attached to
the carboxy group of the R group of any Asp and Glu residue.
[0134] In another embodiment, a lipophilic substituent is attached
to the carboxy group attached to the alpha-carbon of the C-terminal
amino acid.
[0135] In a most preferred embodiment, a lipophilic substituent is
attached to the epsilon-amino group of any Lys residue.
[0136] Each lipophilic substituent contains a functional group
which may be attached to a functional group of an amino acid of the
parent GLP-1 peptide. For example, a lipophilic substituent may
contain a carboxyl group which can be attached to an amino group of
the parent GLP-1 peptide by means of an amide bond.
[0137] In an embodiment, the lipophilic substituent comprises a
partially or completely hydrogenated cyclopentanophenathrene
skeleton.
[0138] In another embodiment, the lipophilic substituent is a
straight-chain or branched alkyl group.
[0139] In another embodiment, the lipophilic substituent is an acyl
group of a straight-chain or branched fatty acid. Preferably, the
lipophilic substituent is an acyl group having the formula
CH.sub.3(CH.sub.2).sub.nC- O--, wherein n is an integer from 4 to
38, preferably an integer from 12 to 38, and most preferably is
CH.sub.3(CH.sub.2).sub.12CO--, CH.sub.3(CH.sub.2).sub.14CO--,
CH.sub.3(CH.sub.2).sub.16CO--, CH.sub.3(CH.sub.2).sub.18CO--,
CH.sub.3(CH.sub.2).sub.20CO-- and CH.sub.3(CH.sub.2).sub.22CO--. In
a more preferred embodiment, the lipophilic substituent is
tetradecanoyl. In a most preferred embodiment, the lipophilic
substituent is hexadecanoyl.
[0140] In another embodiment of the present invention, the
lipophilic substituent has a group which is negatively charged such
as a carboxylic acid group. For example, the lipophilic substituent
may be an acyl group of a straight-chain or branched alkane
.alpha.,.omega.-dicarboxylic acid of the formula
HOOC(CH.sub.2).sub.mCO--, wherein m is an integer from 4 to 38,
preferably an integer from 12 to 38, and most preferably is
HOOC(CH.sub.2).sub.14CO--, HOOC(CH.sub.2).sub.16CO--,
HOOC(CH.sub.2).sub.18CO--, HOOC(CH.sub.2).sub.20CO-- or
HOOC(CH.sub.2).sub.22CO--.
[0141] In a preferred embodiment of the invention, the lipophilic
substituent is attached to the parent GLP-1 peptide by means of a
spacer. A spacer must contain at least two functional groups, one
to attach to a functional group of the lipophilic substituent and
the other to a functional group of the parent GLP-1 peptide.
[0142] In an embodiment, the spacer is an amino acid residue except
Cys or Met, or a dipeptide such as Gly-Lys. For purposes of the
present invention, the phrase "a dipeptide such as Gly-Lys" means
any combination of two amino acids except Cys or Met, preferably a
dipeptide wherein the C-terminal amino acid residue is Lys, His or
Trp, preferably Lys, and the N-terminal amino acid residue is Ala,
Arg, Asp, Asn, Gly, Glu, Gln, Ile, Leu, Val, Phe, Pro, Ser, Tyr,
Thr, Lys, His and Trp. Preferably, an amino group of the parent
peptide forms an amide bond with a carboxylic group of the amino
acid residue or dipeptide spacer, and an amino group of the amino
acid residue or dipeptide spacer forms an amide bond with a
carboxyl group of the lipophilic substituent.
[0143] Preferred spacers are lysyl, glutamyl, asparagyl, glycyl,
beta-alanyl and gamma-aminobutanoyl, each of which constitutes an
individual embodiment. Most preferred spacers are glutamyl and
beta-alanyl. When the spacer is Lys, Glu or Asp, the carboxyl group
thereof may form an amide bond with an amino group of the amino
acid residue, and the amino group thereof may form an amide bond
with a carboxyl group of the lipophilic substituent. When Lys is
used as the spacer, a further spacer may in some instances be
inserted between the .epsilon.-amino group of Lys and the
lipophilic substituent. In one embodiment, such a further spacer is
succinic acid which forms an amide bond with the .epsilon.-amino
group of Lys and with an amino group present in the lipophilic
substituent. In another embodiment such a further spacer is Glu or
Asp which forms an amide bond with the .epsilon.-amino group of Lys
and another amide bond with a carboxyl group present in the
lipophilic substituent, that is, the lipophilic substituent is a
N.sup..epsilon.-acylated lysine residue.
[0144] In another embodiment, the spacer is an unbranched alkane
.alpha.,.omega.-dicarboxylic acid group having from 1 to 7
methylene groups, which spacer forms a bridge between an amino
group of the parent peptide and an amino group of the lipophilic
substituent. Preferably, the spacer is succinic acid.
[0145] In a further embodiment, the lipophilic substituent with the
attached spacer is a group of the formula
CH.sub.3(CH.sub.2).sub.pNH--CO(- CH.sub.2).sub.qCO--, wherein p is
an integer from 8 to 33, preferably from 12 to 28 and q is an
integer from 1 to 6, preferably 2.
[0146] In a further embodiment, the lipophilic substituent with the
attached spacer is a group of the formula
CH.sub.3(CH.sub.2).sub.rCO--NHC- H(COOH)(CH.sub.2).sub.2CO--,
wherein r is an integer from 4 to 24, preferably from 10 to 24.
[0147] In a further embodiment, the lipophilic substituent with the
attached spacer is a group of the formula
CH.sub.3(CH.sub.2).sub.sCO--NHC- H((CH.sub.2).sub.2COOH)CO--,
wherein s is an integer from 4 to 24, preferably from 10 to 24.
[0148] In a further embodiment, the lipophilic substituent is a
group of the formula COOH(CH.sub.2).sub.tCO-- wherein t is an
integer from 6 to 24.
[0149] In a further embodiment, the lipophilic substituent with the
attached spacer is a group of the formula
--NHCH(COOH)(CH.sub.2).sub.4NH-- -CO(CH.sub.2).sub.uCH.sub.3,
wherein u is an integer from 8 to 18.
[0150] In a further embodiment, the lipophilic substituent with the
attached spacer is a group of the formula
CH.sub.3(CH.sub.2).sub.vCO--NH-- -(CH.sub.2).sub.z--CO, wherein v
is an integer from 4 to 24 and z is an integer from 1 to 6.
[0151] In a further embodiment, the lipophilic substituent with the
attached spacer is a group of the formula
--NHCH(COOH)(CH.sub.2).sub.4NH--
-COCH((CH.sub.2).sub.2COOH)NH--CO(CH.sub.2).sub.wCH.sub.3, wherein
w is an integer from 10 to 16.
[0152] In a further embodiment, the lipophilic substituent with the
attached spacer is a group of the formula
--NHCH(COOH)(CH.sub.2).sub.4NH--
-CO(CH.sub.2).sub.2CH(COOH)NHCO(CH.sub.2).sub.xCH.sub.3, wherein x
is zero or an integer from 1 to 22, preferably 10 to 16.
[0153] In a further embodiment the GLP-1 derivative is derived from
a GLP-1 fragment selected from the group comprising 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) and GLP-1(7-41) or an analogue
thereof.
[0154] In a further embodiment of the GLP-1 derivative the
designation analogue comprises derivatives wherein a total of up to
fifteen, preferably up to ten amino acid residues have been
exchanged with any a-amino acid residue.
[0155] In a further embodiment of the GLP-1 derivative the
designation analogue comprises derivatives wherein a total of up to
fifteen, preferably up to ten amino acid residues have been
exchanged with any a-amino acid residue which can be coded for by
the genetic code.
[0156] In a further embodiment of the GLP-1 derivative the
designation analogue comprises derivatives wherein a total of up to
six amino acid residues have been exchanged with another
.alpha.-amino acid residue which can be coded for by the genetic
code.
[0157] In a further embodiment the GLP-1 derivative is a GLP-1
derivative of formula I:
2 7 8 9 10 11 12 13 14 15 16 17 His-Xaa-Xaa-Gly-Xaa-Phe--
Thr-Xaa-Asp-Xaa-Xaa- 18 19 20 21 22 23 24 25 26 27 28
Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Phe- 29 30 31 32 33 34 35
36 37 38 Ile-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa 39 40 41 42 43 44
45 Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa (I)
[0158] wherein
[0159] Xaa at position 8 is Ala, Gly, Ser, Thr, Leu, Ile, Val, Glu,
Asp, Met, or Lys,
[0160] Xaa at position 9 is Glu, Asp, or Lys,
[0161] Xaa at position 111 is Thr, Ala, Gly, Ser, Leu, Ile, Val,
Glu, Asp, or Lys,
[0162] Xaa at position 14 is Ser, Ala, Gly, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0163] Xaa at position 16 is Val, Ala, Gly, Ser, Thr, Leu, Ile,
Tyr, Glu, Asp, or Lys,
[0164] Xaa at position 17 is Ser, Ala, Gly, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0165] Xaa at position 18 is Ser, Ala, Gly, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0166] Xaa at position 19 is Tyr, Phe, Trp, Glu, Asp, or Lys,
[0167] Xaa at position 20 is Leu, Ala, Gly, Ser, Thr, Leu, Ile,
Val, Glu, Asp, or Lys,
[0168] Xaa at position 21 is Glu, Asp, or Lys,
[0169] Xaa at position 22 is Gly, Ala, Ser, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0170] Xaa at position 23 is Gln, Asn, Arg, Glu, Asp, or Lys,
[0171] Xaa at position 24 is Ala, Gly, Ser, Thr, Leu, Ile, Val,
Arg, Glu, Asp, or Lys,
[0172] Xaa at position 25 is Ala, Gly, Ser, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0173] Xaa at position 26 is Lys, Arg, Gln, Glu, Asp, or His,
[0174] Xaa at position 27 is Glu, Asp, or Lys,
[0175] Xaa at position 30 is Ala, Gly, Ser, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0176] Xaa at position 31 is Trp, Phe, Tyr, Glu, Asp, or Lys,
[0177] Xaa at position 32 is Leu, Gly, Ala, Ser, Thr, Ile, Val,
Glu, Asp, or Lys,
[0178] Xaa at position 33 is Val, Gly, Ala, Ser, Thr, Leu, Ile,
Glu, Asp, or Lys,
[0179] Xaa at position 34 is Lys, Arg, Glu, Asp, or His,
[0180] Xaa at position 35 is Gly, Ala, Ser, Thr, Leu, Ile, Val,
Glu, Asp, or Lys,
[0181] Xaa at position 36 is Arg, Lys, Glu, Asp, or His,
[0182] Xaa at position 37 is Gly, Ala, Ser, Thr, Leu, Ile, Val,
Glu, Asp, or Lys, or is deleted,
[0183] Xaa at position 38 is Arg, Lys, Glu, Asp, or His, or is
deleted,
[0184] Xaa at position 39 is Arg, Lys, Glu, Asp, or His, or is
deleted,
[0185] Xaa at position 40 is Asp, Glu, or Lys, or is deleted,
[0186] Xaa at position 41 is Phe, Trp, Tyr, Glu, Asp, or Lys, or is
deleted,
[0187] Xaa at position 42 is Pro, Lys, Glu, or Asp, or is
deleted,
[0188] Xaa at position 43 is Glu, Asp, or Lys, or is deleted,
[0189] Xaa at position 44 is Glu, Asp, or Lys, or is deleted,
and
[0190] Xaa at position 45 is Val, Glu, Asp, or Lys, or is deleted,
or
[0191] (a) a C-1-6-ester thereof, (b) amide, C-1-6-alkylamide, or
C-1-6-dialkylamide thereof and/or (c) a pharmaceutically acceptable
salt thereof,
[0192] provided that
[0193] (i) when the amino acid at position 37, 38, 39, 40, 41, 42,
43 or 44 is deleted, then each amino acid downstream of the amino
acid is also deleted,
[0194] (ii) the derivative of the GLP-1 analog contains only one or
two Lys,
[0195] (iii) the .epsilon.-amino group of one or both Lys is
substituted with a lipophilic substituent optionally via a
spacer,
[0196] (iv) the total number of different amino acids between the
derivative of the GLP-1 analog and the corresponding native form of
GLP-1 does not exceed six.
[0197] In a further embodiment of the GLP-1 derivative of formula
I, the amino acids at positions 37-45 are absent.
[0198] In another embodiment of the GLP-1 derivative of formula I,
the amino acids at positions 38-45 are absent.
[0199] In another embodiment of the GLP-1 derivative of formula I,
the amino acids at positions 3945 are absent.
[0200] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 8 is Ala, Gly, Ser, Thr, Leu, Ile, Val, Glu, Asp,
or Lys,
[0201] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 8 is Ala, Gly, Ser, Thr, or Val.
[0202] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 9 is Glu.
[0203] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position II is Thr.
[0204] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 14 is Ser.
[0205] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 16 is Val.
[0206] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 17 is Ser.
[0207] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 18 is Ser, Lys, Glu, or Asp.
[0208] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 19 is Tyr, Lys, Glu, or Asp.
[0209] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 20 is Leu, Lys, Glu, or Asp.
[0210] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 21 is Glu, Lys, or Asp.
[0211] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 22 is Gly, Glu, Asp, or Lys.
[0212] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 23 is Gln, Glu, Asp, or Lys.
[0213] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 24 is Ala, Glu, Asp, or Lys.
[0214] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 25 is Ala, Glu, Asp, or Lys.
[0215] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 26 is Lys, Glu, Asp, or Arg.
[0216] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 27 is Glu, Asp, or Lys.
[0217] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 30 is Ala, Glu, Asp, or Lys.
[0218] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 31 is Trp, Glu, Asp, or Lys.
[0219] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 32 is Leu, Glu, Asp, or Lys.
[0220] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 33 is Val, Glu, Asp, or Lys.
[0221] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 34 is Lys, Arg, Glu, or Asp.
[0222] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 35 is Gly, Glu, Asp, or Lys.
[0223] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 36 is Arg, Lys, Glu, or Asp.
[0224] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 37 is Gly, Glu, Asp, or Lys.
[0225] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 38 is Arg, or Lys, or is deleted.
[0226] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 39 is deleted.
[0227] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 40 is deleted.
[0228] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 41 is deleted.
[0229] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 42 is deleted.
[0230] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 43 is deleted.
[0231] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 44 is deleted.
[0232] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 45 is deleted.
[0233] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 26 is Arg, each of Xaa at positions 37-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-36).
[0234] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 26 is Arg, each of Xaa at positions 38-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-37).
[0235] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 26 is Arg, each of Xaa at positions 39-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-38).
[0236] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 34 is Arg, each of Xaa at positions 37-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-36).
[0237] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 34 is Arg, each of Xaa at positions 38-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-37).
[0238] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 34 is Arg, each of Xaa at positions 39-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-38).
[0239] In another embodiment of the GLP-1 derivative of formula I,
Xaa at positions 26 and 34 is Arg, Xaa at position 36 is Lys, each
of Xaa at positions 37-45 is deleted, and each of the other Xaa is
the amino acid in native GLP-1(7-36).
[0240] In another embodiment of the GLP-1 derivative of formula I,
Xaa at positions 26 and 34 is Arg, Xaa at position 36 is Lys, each
of Xaa at positions 38-45 is deleted, and each of the other Xaa is
the amino acid in native GLP-1(7-37).
[0241] In another embodiment of the GLP-1 derivative of formula I,
Xaa at positions 26 and 34 is Arg, Xaa at position 36 is Lys, each
of Xaa at positions 39-45 is deleted, and each of the other Xaa is
the amino acid in native GLP-1(7-38).
[0242] In another embodiment of the GLP-1 derivative of formula I,
Xaa at positions 26 and 34 is Arg, Xaa at position 38 is Lys, each
of Xaa at positions 39-45 is deleted, and each of the other Xaa is
the amino acid in native GLP-1(7-38).
[0243] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 8 is Thr, Ser, Gly or Val, Xaa at position 37 is
Glu, Xaa at position 36 is Lys, each of Xaa at positions 38-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-37).
[0244] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 8 is Thr, Ser, Gly or Val, Xaa at position 37 is
Glu, Xaa at position 36 is Lys, each of Xaa at positions 39-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-38).
[0245] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 8 is Thr, Ser, Gly or Val, Xaa at position 37 is
Glu, Xaa at position 38 is Lys, each of Xaa at positions 39-45 is
deleted, and each of the other Xaa is the amino acid in native
GLP-1(7-38).
[0246] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 18, 23 or 27 is Lys, and Xaa at positions 26 and 34
is Arg, each of Xaa at positions 37-45 is deleted, and each of the
other Xaa is the amino acid in native GLP-1(7-36).
[0247] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 18, 23 or 27 is Lys, and Xaa at positions 26 and 34
is Arg, each of Xaa at positions 38-45 is deleted, and each of the
other Xaa is the amino acid in native GLP-1(7-37).
[0248] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 18, 23 or 27 is Lys, and Xaa at positions 26 and 34
is Arg, each of Xaa at positions 39-45 is deleted, and each of the
other Xaa is the amino acid in native GLP-1(7-38).
[0249] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 8 is Thr, Ser, Gly, or Val, Xaa at position 18, 23
or 27 is Lys, and Xaa at position 26 and 34 is Arg, each of Xaa at
positions 37-45 is deleted, and each of the other Xaa is the amino
acid in native GLP-1(7-36).
[0250] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 8 is Thr, Ser, Gly, or Val, Xaa at position 18, 23
or 27 is Lys, and Xaa at position 26 and 34 is Arg, each of Xaa at
positions 3845 is deleted, and each of the other Xaa is the amino
acid in native GLP-1(7-37).
[0251] In another embodiment of the GLP-1 derivative of formula I,
Xaa at position 8 is Thr, Ser, Gly, or Val, Xaa at position 18, 23
or 27 is Lys, and Xaa at position 26 and 34 is Arg, each of Xaa at
positions 39-45 is deleted, and each of the other Xaa is the amino
acid in native GLP-1(7-38).
[0252] Such GLP-1 derivatives includes, but is not limited to,
[0253]
Lys.sup.34(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..alpha.-tetradec-
anoyl))) GLP-1 (7-37),
[0254]
Arg.sup.26,34,Lys.sup.8(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..al-
pha.-hexadecanoyl))) GLP-1 (7-37),
[0255]
Arg.sup.34,Lys.sup.26(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..alph-
a.-dodecanoyl))) GLP-1 (7-37),
[0256]
Arg.sup.34,Lys.sup.26(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha.--
hexadecanoyl))) GLP-1 (7-37),
[0257]
Arg.sup.34,Lys.sup.26(N.sup..epsilon.-(.alpha.-glutamyl(N.sup..alph-
a.-hexadecanoyl))) GLP-1 (7-37),
[0258]
Arg.sup.34,Lys.sup.26(N.sup..epsilon.-(piperidinyl-4-carbonyl(N-hex-
adecanoyl))) GLP-1 (7-37),
[0259]
Arg.sup.34,Lys.sup.26(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..alph-
a.-decanoyl))) GLP-1 (7-37),
[0260]
Glu.sup.22,23,30Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.gamma.-gl-
utamyl(N.sup..alpha.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0261]
Glu.sup.23,26Arg.sup.34Lys.sup.38(N.sup..epsilon.-(.gamma.-glutamyl-
(N.sup..alpha.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0262]
Lys.sup.26,34-bis(N.sup..epsilon.-(7-glutamyl(N.sup..alpha.-tetrade-
canoyl)))-GLP-1(7-37)-OH,
[0263]
Lys.sup.26,34-bis(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..alpha.-h-
exadecanoyl)))-GLP-1(7-37)-OH,
[0264]
Arg.sup.34Lys.sup.26(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..alpha-
.-hexadecanoyl)))-GLP-1(7-37)-OH,
[0265]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..al-
pha.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0266]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..al-
pha.-hexadecanoyl)))-GLP-1(7-38)-OH,
[0267]
Arg.sup.34Lys.sup.26(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..alpha-
.-tetradecanoyl)))-GLP-1(7-37)-OH,
[0268]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..al-
pha.-octadecanoyl)))-GLP-1(7-38)-OH.
[0269]
Glu.sup.22,23,30Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.gamma.-gl-
utamyl(N.sup..alpha.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0270]
Glu.sup.23,26Arg.sup.34Lys.sup.38(N.sup..epsilon.-(.gamma.-glutamyl-
(N.sup..alpha.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0271]
Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxytridecanoyl))-GLP-
-1(7-37)-OH,
[0272]
Lys.sup.26,34-bis(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..alpha.-t-
etradecanoyl)))-GLP-1(7-37)-OH,
[0273]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.omega.-carboxypentadecano-
yl))-GLP-1(7-38)-OH,
[0274]
Lys.sup.26,34-bis(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..alpha.-h-
exadecanoyl)))-GLP-1(7-37)-OH,
[0275]
Arg.sup.34Lys.sup.26(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..alpha-
.-hexadecanoyl)))-GLP-1(7-37)-OH,
[0276]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..al-
pha.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0277]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.omega.-carboxypentadecano-
yl))-GLP-1(7-38)-OH,
[0278]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..al-
pha.-hexadecanoyl)))-GLP-1(7-38)-OH,
[0279]
Arg.sup.18,23,26,30,34Lys.sup.38(N.sup..epsilon.-hexadecanoyl)-GLP--
1(7-38)-OH,
[0280]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.omega.-carboxytridecanoyl-
))-GLP-1(7-38)-OH,
[0281]
Arg.sup.34Lys.sup.26(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..alpha-
.-tetradecanoyl)))-GLP-1(7-37)-OH,
[0282]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.gamma.-glutamyl(N.sup..al-
pha.-octadecanoyl)))-GLP-1(7-38)-OH,
[0283]
Glu.sup.22,23,30Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.beta.-ala-
nyl(N.sup..alpha.-tetra decanoyl)))-GLP-1(7-38)-OH,
[0284]
Glu.sup.23,26Arg.sup.34Lys.sup.38(N.sup..epsilon.-(.beta.-alanyl(N.-
sup..alpha.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0285]
Lys.sup.26,34-bis(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha.-tetr-
adecanoyl)))-GLP-1(7-37)-OH,
[0286]
Lys.sup.26,34-bis(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha.-hexa-
decanoyl)))-GLP-1(7-37)-OH,
[0287]
Arg.sup.34Lys.sup.26(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha.-h-
exadecanoyl)))-GLP-1(7-37)-OH,
[0288]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha-
.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0289]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha-
.-hexadecanoyl)))-GLP-1(7-38)-OH,
[0290]
Arg.sup.34Lys.sup.26(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha.-t-
etradecanoyl)))-GLP-1(7-37)-OH,
[0291]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha-
.-octadecanoyl)))-GLP-1(7-38)-OH.
[0292]
Glu.sup.22,23,30Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.beta.-ala-
nyl(N.sup..alpha.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0293]
Glu.sup.23,26Arg.sup.34Lys.sup.38(N.sup..epsilon.-(.beta.-alanyl(N.-
sup..alpha.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0294]
Lys.sup.26,34-bis(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha.-tetr-
adecanoyl)))-GLP-1(7-37)-OH,
[0295]
Lys.sup.26,34-bis(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha.-hexa-
decanoyl)))-GLP-1(7-37)-OH,
[0296]
Arg.sup.34Lys.sup.26(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha.-h-
exadecanoyl)))-GLP-1(7-37)-OH,
[0297]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha-
.-tetradecanoyl)))-GLP-1(7-38)-OH,
[0298]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha-
.-hexadecanoyl)))-GLP-1(7-38)-OH,
[0299]
Arg.sup.34Lys.sup.25(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha.-t-
etradecanoyl)))-GLP-1(7-37)-OH,
[0300]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(.beta.-alanyl(N.sup..alpha-
.-octadecanoyl)))-GLP-1(7-38)-OH,
[0301] Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-37);
[0302] Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-37);
[0303]
Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-37);
[0304]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-37);
[0305]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-37);
[0306]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-3-
7);
[0307]
Val.sup.8Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-37);
[0308]
Val.sup.8Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-37);
[0309]
Val.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-3-
7);
[0310]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-37);
[0311] Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-38);
[0312] Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-38);
[0313]
Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-38);
[0314]
Gly.sup.8Lys.sup.26-(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-38);
[0315]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-38);
[0316]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-3-
8);
[0317]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-38);
[0318] Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-39);
[0319] Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-39);
[0320]
Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-39);
[0321]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-39);
[0322]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-39);
[0323]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-3-
9);
[0324]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-39);
[0325] Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-40);
[0326] Lys.sup.34(N.sup..epsilon.-tradecanoyl)-GLP-1(7-40);
[0327]
Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-40);
[0328]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-40);
[0329]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-40);
[0330]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-4-
0);
[0331] Arg.sup.26Lys.sup.2-tetradecanoyl)-GLP-1(7-40);
[0332] Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36);
[0333] Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36);
[0334]
Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36);
[0335]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36);
[0336]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36);
[0337]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-3-
6);
[0338]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36);
[0339] Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-35);
[0340] Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-35);
[0341]
Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-35);
[0342]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-35);
[0343]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-35));
[0344]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-3-
5);
[0345]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-35);
[0346]
Lys.sup.26(N.sup..alpha.-tetradecanoyl)-GLP-1(7-36)amide;
[0347]
Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36)amide;
[0348]
Lys.sup.26,34-bis(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36)amide;
[0349]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36)amide-
;
[0350]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36)amide-
;
[0351]
Gly.sup.8Lys.sup.26,34-bis(N.sup..alpha.-tetradecanoyl)-GLP-1(7-36)-
amide;
[0352]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-36)amid-
e;
[0353]
Gly.sup.8Arg.sup.26Lys.sup.34-tetradecanoyl)-GLP-1(7-37);
[0354]
Lys.sup.26(N.sup..epsilon.-tetradecanoyl)Arg.sup.34-GLP-1(7-37);
[0355]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-tetradecanoyl)Arg.sup.34-GLP-1(-
7-37);
[0356]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-37);
[0357]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-tetradecanoyl)-GLP-
-1(7-37);
[0358]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(-
7-38);
[0359] Arg.sup.26,34 1
[0360]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-tetradecanoyl)-GLP-
-1(7-38);
[0361]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-tetradecanoyl)-GLP-1(-
7-39);
[0362]
Lys.sup.26(N.sup..epsilon.-tetradecanoyl)Arg.sup.34-GLP-1(7-39);
[0363]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-tetradecanoyl)Arg.sup.34-GLP-1(-
7-39);
[0364]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-tetradecanoyl)-GLP-1(7-39);
[0365]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-tetradecanoyl)-GLP-
-1(7-39);
[0366] Arg.sup.26 Lys.sup.2 (N-tetradecanoyl)-GLP-1(7-34);
[0367]
Lys.sup.26(N.sup..epsilon.-tetradecanoyl)Arg.sup.34-GLP-1(7-40);
[0368] Arg256G34
Lys.sup.36(N.sup..alpha.-tetradecanoyl)Ar3-GLP-1(7-340);
[0369] Gly
Arg.sup.26.sup.34Lys.sup.36(N-tetradecanoyl)-GLP-1(7-30);
[0370] Gly 8Arg.sup.26 Lys.sup.3
(N.sup..alpha.-tetradecanoyl)-GLP-1 (7-40);
[0371] Gly.sup.8Lys.sup.26(N-tetr adeca noyl)Arg-GLP-1(7-39);
[0372] Arg.sup.26,34 Lys.sup.36(N.sup..alpha.-tetradecanoyl)Arg
GLP-1(7-39);
[0373] Gly.sup.8Arg.sup.26,34Lys.sup.36(-tetradecanoyl)-GLP-1
(7-40);
[0374] Lys.sup.26(N'-((crboyadecanoyl)-GLP-1 (7-37);
[0375]
Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1
(7-37);
[0376]
Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GL-
P-1(7-37)(;
[0377]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))--
GLP-1(7-37);
[0378]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))--
GLP-1(7-37);
[0379]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadeca-
noyl))-GLP-1(7-37);
[0380]
Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1(7-3-
8);
[0381]
Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1(7-3-
8);
[0382]
Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GL-
P-1(7-38);
[0383]
Gly.sup.8Lys.sup.26(N'-(.omega.-carboxynonadecanoyl))-GLP-1(7-38);
[0384]
Gly.sup.8Lys.sup.34(N-(.omega.-carboxynonadecanoyl))-GLP-1(7-38);
[0385]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadeca-
noyl))-GLP-1(7-38);
[0386]
Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1(7-3-
9);
[0387]
Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1(7-3-
9);
[0388]
Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GL-
P-1(7-39);
[0389]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))--
GLP-1(7-39);
[0390]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadeca-
noyl))-GLP-1(7-39);
[0391]
Lys.sup.26-(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1(7--
40);
[0392]
Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1(7-4-
0);
[0393]
Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GL-
P-1(7-40);
[0394]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))--
GLP-1(7-40);
[0395]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))--
GLP-1(7-40);
[0396]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadeca-
noyl))-GLP-1(7-40);
[0397] Lys.sup.34(N'-((o-carboxynonadecanoyl))-GLP-1(7-36);
[0398] Lys.sup.26,34-bis(N-((-carboxynonadecanoyl))-GLP 1(743
6);
[0399] Lys.sup.26(N'-(O)-carboxynonadecanoyl))-GLP-1(7-36);
[0400] Gly Lys.sup.34(N-(6)-carboxynonadecanoyl))-GLP-1 (7-36);
[0401]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadeca-
noyl))-GLP-1(7-36);
[0402]
Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1(7-3-
6)amide;
[0403]
Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1(7-3-
6)amide;
[0404]
Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GL-
P-1(7-36)amide;
[0405]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))--
GLP-1(7-36)amide;
[0406]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))--
GLP-1(7-36)amide;
[0407]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadeca-
noyl))-GLP-1(7-36)amide;
[0408]
Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1(7-3-
5);
[0409]
Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GLP-1(7-3-
5);
[0410]
Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-GL-
P-1(7-35);
[0411]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))--
GLP-1(7-35);
[0412]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))--
GLP-1(7-35);
[0413]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(.omega.-carboxynonadeca-
noyl))-GLP-1(7-35);
[0414]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-
-GLP-1(7-37);
[0415]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonad-
ecanoyl))-GLP-1(7-37);
[0416] Gly sA26
(NL-(No-carboxynonadecanoyl))Arg.sup.34-GLP-1(7-37);
[0417] G26,34(N'-(-carboxynonadecanoyl))A GLP-1(7-37);
[0418]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(.omega.-carboxynonadecanoy-
l))-GLP-1(7-37);
[0419] 8Arg.sup.26Lys.sup.36((N-(o-carboxynonadecanoyl))-GLP-1 (7-3
74;
[0420] GyArg.sup.26Lys.sup.34(N'-(co-carboxynonadecanoyl))-GLP-1
(7-38);
[0421] Gl Ar 6Lys.sup.34(N-(co-carboxynonadecanoyl))Ar3-GLP-1
(7-38);
[0422]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))A-
rg.sup.34-GLP-1(7-38);
[0423] Arg.sup.26,34(o36)-carboxynonadecanoyl))-GLP-1(7-38);
[0424]
Arg.sup.26,34Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoy-
l))-GLP-1(7-38);
[0425] A8 g26,34LyS38(N'-((o-carboxynonadecanoyl))-GLP-1
(7-38);
[0426]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-
-GLP-1(7-39);
[0427]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonad-
ecanoyl))-GLP-1(7-39);
[0428]
Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))Arg.sup.34-
-GLP-1(7-39);
[0429]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))A-
rg.sup.34-GLP-1(7-39);
[0430]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(.omega.-carboxynonadecanoy-
l))-GLP-1(7-39);
[0431]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(.omega.-carboxyno-
nadecanoyl))-GLP-1(7-39);
[0432]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))-
-GLP-1(7-40);
[0433]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(.omega.-carboxynonad-
ecanoyl))-GLP-1(7-40);
[0434]
Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))Arg.sup.34-
-GLP-1(7-40);
[0435]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(.omega.-carboxynonadecanoyl))A-
rg.sup.34-GLP-1(7-40);
[0436]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(.omega.-carboxynonadecanoy-
l))-GLP-1(7-40);
[0437]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(.omega.-carboxyno-
nadecanoyl))-GLP-1(7-40);
[0438]
Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-37);
[0439]
Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-37);
[0440]
Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-37);
[0441]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-37);
[0442]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-37);
[0443]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(-
7-37);
[0444]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-37);
[0445]
Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-38);
[0446]
Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-38);
[0447]
Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-38);
[0448]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-38);
[0449]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-38);
[0450]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(-
7-38);
[0451]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-38);
[0452]
Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-39);
[0453]
Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-39);
[0454]
Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-39);
[0455]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-39);
[0456]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-39);
[0457]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(-
7-39);
[0458]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-39);
[0459]
Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-40);
[0460]
Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-40);
[0461]
Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-40);
[0462]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-40);
[0463]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-40);
[0464]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(-
7-40);
[0465]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-40);
[0466]
Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-36);
[0467]
Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-36);
[0468]
Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-36);
[0469]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-36);
[0470]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-36);
[0471]
Gly.sup.8Lys.sup.26,34-bis-(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1-
(7-36);
[0472]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-36);
[0473]
Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-35);
[0474]
Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-35);
[0475]
Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-35);
[0476]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-35);
[0477]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-35);
[0478]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(-
7-35);
[0479]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-35);
[0480]
Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-36)amide;
[0481]
Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-36)amide;
[0482]
Lys.sup.26,34-bis(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-36)amid-
e;
[0483]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-37)am-
ide 1s6(N,-(7-deoxycholoyl))-GLP-1(7-36)amide;
[0484]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-36);
[0485]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-37);
[0486]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(7-deoxycholoyl))--
GLP-1(7-37);
[0487] Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-37);
[0488] Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-37);
[0489]
Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-37);
[0490]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-37);
[0491]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-37);
[0492]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-37);
[0493]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-37);
[0494]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-
-1(7-38);
[0495]
Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))Arg.sup.34-GLP-1(7-38);
[0496]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))Arg.sup.34-GLP-
-1(7-38);
[0497]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-3-
8);
[0498]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-3-
8);
[0499]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(7-deoxycholoyl))--
GLP-1(7-38);
[0500] Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-38);
[0501] Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-38);
[0502]
Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-38);
[0503]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-38);
[0504]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-38);
[0505]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-38);
[0506]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-38);
[0507]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-
-1(7-39);
[0508]
Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))Arg.sup.34-GLP-1(7-39);
[0509]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))Arg.sup.34-GLP-
-1(7-39);
[0510]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-3-
9);
[0511]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(7-deoxycholoyl))--
GLP-1(7-39);
[0512] Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-39);
[0513] Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-39);
[0514]
Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-39);
[0515]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-39);
[0516]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-39);
[0517]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-39);
[0518]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-39);
[0519]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(7-deoxycholoyl))-GLP-
-1(7-40);
[0520]
Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))Arg.sup.34-GLP-1(7-40);
[0521]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(7-deoxycholoyl))Arg.sup.34-GLP-
-1(7-40);
[0522]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(7-deoxycholoyl))-GLP-1(7-4-
0);
[0523]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(7-deoxycholoyl))--
GLP-1(7-40);
[0524] Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-40);
[0525] Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-40);
[0526]
Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-40);
[0527]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-40);
[0528]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-40);
[0529]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-40);
[0530]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-40);
[0531] Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-36);
[0532] Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-36);
[0533]
Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-36);
[0534]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-36);
[0535]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-36);
[0536]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-36);
[0537]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-36);
[0538] Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-35);
[0539] Lys.sup.34(N.sup..epsilon.-(choloyl)-GLP-1(7-35);
[0540]
Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-35);
[0541]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-35);
[0542]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-35);
[0543]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-35);
[0544]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-35);
[0545] Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-36)amide;
[0546] Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-36)amide;
[0547]
Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl)-GLP-1(7-36)amide;
[0548]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))-GLP-1(7-36)amide;
[0549]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-36)amide;
[0550]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(choloyl))-GLP-1(7-36)am-
ide;
[0551]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-36)amide;
[0552] Gly
8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-37);
[0553]
Lys.sup.26(N.sup..epsilon.-(choloyl))Arg.sup.34-GLP-1(7-37);
[0554]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))Arg.sup.34-GLP-1(7-37-
);
[0555]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(choloyl))-GLP-1(7-37);
[0556]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(choloyl))-GLP-1(7-
-37);
[0557] Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-37);
[0558] Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-37);
[0559]
Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-37);
[0560]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-37);
[0561]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-37);
[0562]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7--
37);
[0563]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-37);
[0564]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-38-
);
[0565]
Lys.sup.26(N.sup..epsilon.-(choloyl))Arg.sup.34-GLP-1(7-38);
[0566]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))Arg.sup.34-GLP-1(7-38-
);
[0567]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(choloyl))-GLP-1(7-38);
[0568]
Arg.sup.26,34Lys.sup.3(N.sup..epsilon.-(choloyl))-GLP-1(7-38);
[0569]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(choloyl))-GLP-1(7-
-38);
[0570] Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-38);
[0571] Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-38);
[0572]
Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-38);
[0573]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-38);
[0574]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-38);
[0575]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7--
38);
[0576]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-38);
[0577]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-39-
);
[0578]
Lys.sup.26(N.sup..epsilon.-(choloyl))Arg.sup.34-GLP-1(7-39);
[0579]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))Arg.sup.34-GLP-1(7-39-
);
[0580]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(choloyl))-GLP-1(7-39);
[0581]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(choloyl))-GLP-1(7-
-39);
[0582] Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-39);
[0583] Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-39);
[0584]
Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-39);
[0585]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-39);
[0586]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-39);
[0587]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7--
39);
[0588]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-39);
[0589]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(choloyl))-GLP-1(7-40-
);
[0590]
Lys.sup.26(N.sup..epsilon.-(choloyl))Arg.sup.34-GLP-1(7-40);
[0591]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(choloyl))Arg.sup.34-GLP-1(7-40-
);
[0592]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(choloyl))-GLP-1(7-40);
[0593]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(choloyl))-GLP-1(7-
-40);
[0594] Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-40);
[0595] Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-40);
[0596]
Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-40);
[0597] AGl LS26 y3(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-40);
[0598]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-40);
[0599]
Lys26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-33);
[0600]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-36);
[0601]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-36);
[0602]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7--
36);
[0603]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-36);
[0604] Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-35);
[0605] Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-35);
[0606]
Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-35);
[0607]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-35);
[0608]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-35);
[0609]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7--
35);
[0610]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-35);
[0611]
Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-36)amide;
[0612]
Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-36)amide;
[0613]
Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-36)amide;
[0614]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-36)amid-
e;
[0615]
Gly.sup.8Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-36)amid-
e;
[0616]
Gly.sup.8Lys.sup.26,34-bis(N.sup..epsilon.-(lithocholoyl))-GLP-1(7--
36)amide;
[0617]
Arg.sup.26Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-36)ami-
de;
[0618]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1-
(7-37);
[0619]
Lys.sup.26(N.sup..epsilon.-(lithocholoyl))Arg.sup.34-GLP-1(7-37);
[0620]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(lithocholoyl))Arg.sup.34-GLP-1-
(7-37);
[0621]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-37)-
;
[0622]
Arg.sup.26,34Lys.sup.38(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-37)-
;
[0623] Arg.sup.26Lys.sup.3
(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-38);
[0624] 8y 26 (N'(Il thocholoyrg4-GLP-1(7-38);
[0625] Gly Arg Lys.sup.3(Ilthoc hol loyArg-GLP-1(7-38);
[0626] 2
gyArg.sup.26Lys.sup.3(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-38)-
;
[0627] Ag234Lys.sup.38N'(lith thochloyl))Ar-GLP-1(7-39);
[0628]
Lys.sup.26(N.sup..epsilon.-(lithocholoyl))Arg.sup.34-GLP-1(7-39);
[0629]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-39)-
;
[0630]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(lithocholoyl))-GL-
P-1(7-39);
[0631]
Gly.sup.8Arg.sup.26Lys.sup.34(N.sup..epsilon.-(lithocholoyl))-GLP-1-
(7-40);
[0632]
Lys.sup.26(N.sup..epsilon.-(lithocholoyl))Arg.sup.34-GLP-1(7-40);
[0633]
Gly.sup.8Lys.sup.26(N.sup..epsilon.-(lithocholoyl))Arg.sup.34-GLP-1-
(7-40);
[0634]
Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(lithocholoyl))-GLP-1(7-40)
and
[0635]
Gly.sup.8Arg.sup.26,34Lys.sup.36(N.sup..epsilon.-(lithocholoyl))-GL-
P-1(7-40). Each one of these specific GLP-1 derivatives constitutes
an alternative embodiment of the invention.
[0636] The most preferred GLP-1 derivative is Arg.sup.34,
Lys.sup.26(N.sup..epsilon.-(.gamma.-Glu(N.sup..alpha.-hexadecanoyl)))-GLP-
-1(7-37).
[0637] In a further embodiment of the GLP-1 derivative, a parent
peptide for a derivative of the invention is
3 Arg.sup.26-GLP-1(7-37); Arg.sup.34-GLP-1(7-37);
Lys.sup.36-GLP-1(7-37); Arg.sup.26,34Lys.sup.36-GLP-1(7-37);
Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Arg.sup.26,34Lys.sup.39-GLP-1(7-3- 9);
Arg.sup.26,34Lys.sup.40-GLP-1(7-40);
Arg.sup.26Lys.sup.36-GLP-1(7-37); Arg.sup.34Lys.sup.36-GLP-1(7-37);
Arg.sup.26Lys.sup.39-GLP-1(7-39); Arg.sup.34Lys.sup.40-GL-
P-1(7-40); Arg.sup.26,34Lys.sup.36,39-GLP-1(7-39);
Arg.sup.26,34Lys.sup.36- ,40GLP-1(7-40);
Gly.sup.8Arg.sup.26-GLP-1(7-37); Gly.sup.8Arg.sup.34-GLP-1(7-37);
Gly.sup.8Lys.sup.36-GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Gly.sup.8Arg.sup.26,34Lys.s- up.40-GLP-1(7-40);
Gly.sup.8Arg.sup.26Lys.sup.36-GLP-1(7-3- 7);
Gly.sup.8Arg.sup.34Lys.sup.36-GLP-1(7-37);
Gly.sup.8Arg.sup.26Lys.sup.- 38-GLP-1(7-39);
Gly.sup.8Arg.sup.34Lys.sup.40-GLP-1(7-40);
Gly.sup.8Arg.sup.26,34Lys.sup.36,39-GLP-1(7-39);
Gly.sup.8Arg.sup.26,34Lys.sup.36,40-GLP-1(7-40);
Val.sup.8Arg.sup.34-GLP-- 1(7-37); Val.sup.8Lys.sup.36-GLP-1(7-37);
Val.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7-37);
Val.sup.8Arg.sup.26,34Lys.s- up.39-GLP-1(7-39);
Val.sup.8Arg.sup.26,34Lys.sup.40-GLP-1(- 7-40);
Val.sup.8Arg.sup.26Lys.sup.36-GLP-1(7-37);
Val.sup.8Arg.sup.34Lys.s- up.36-GLP-1(7-37);
Val.sup.8Arg.sup.26Lys.sup.39-GLP-1(7-3- 9);
Val.sup.8Arg.sup.34Lys.sup.40-GLP-1(7-40);
Val.sup.8Arg.sup.26,34Lys.s- up.36,39-GLP-1(7-39); or
Val.sup.8Arg.sup.26,34Lys.sup.36,- 40-GLP-1(7-40).
[0638] In a further embodiment of the GLP-1 derivative, a parent
peptide for a derivative of the invention is:
4 Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Arg.sup.26,34Lys.sup.39GLP-1(- 7-39);
Arg.sup.26,34Lys.sup.40GLP-1(7-40);
Arg.sup.26,34Lys.sup.41GLP-1(7-41);
Arg.sup.26,34Lys.sup.42GLP-1(7-42);
Arg.sup.26,34Lys.sup.43GLP-1(7-43);
Arg.sup.26,34Lys.sup.44GLP-1(7-44);
Arg.sup.26,34Lys.sup.45GLP-1(7-45);
Arg.sup.26Lys.sup.38GLP-1(7-38); Arg.sup.34Lys.sup.38GLP-1(7-38);
Arg.sup.26,34Lys.sup.36,38GLP-1(7-38); Arg.sup.26,34Lys.sup.38GLP-
-1(7-38); Arg.sup.26Lys.sup.39GLP-1(1-39);
Arg.sup.34Lys.sup.39GLP-1(1-39);
Arg.sup.26,34Lys.sup.36,39GLP-1(1-39);
Arg.sup.26Lys.sup.39GLP-1(7-39); Arg.sup.34Lys.sup.39GLP-1(7-39);
Arg.sup.26,34Lys.sup.36,39GLP-1(739).
[0639] In a further embodiment of the GLP-1 derivative, the parent
peptide is selected from the group comprising
Arg.sup.26-GLP-1(7-37), Arg.sup.34-GLP-1(7-37),
Lys.sup.36-GLP-1(7-37), Arg.sup.26,34Lys.sup.36-G- LP-1(7-37),
Arg.sup.26Lys.sup.36-GLP-1(7-37), Arg.sup.34Lys.sup.36-GLP-1(7-
-37), Gly.sup.8Arg.sup.26-GLP-1(7-37),
Gly.sup.8Arg.sup.34-GLP-1(7-37), Gly.sup.8Lys.sup.36-GLP-1(7-37),
Gly.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7- -37),
Gly.sup.8Arg.sup.26Lys.sup.36-GLP-1(7-37) and
Gly.sup.8Arg.sup.34Lys.sup.36-GLP-1(7-37).
[0640] In a further embodiment of the GLP-1 derivative, the parent
peptide is selected from the group comprising
Arg.sup.26Lys.sup.38-GLP-1(7-38),
Arg.sup.26,34Lys.sup.38-GLP-1(7-38),
Arg.sup.26,34Lys.sup.36,38GLP-1(7-38- ),
Gly.sup.8Arg.sup.26Lys.sup.38-GLP-1(7-38) and
Gly.sup.8Arg.sup.26,34Lys- .sup.36,38-GLP-1(7-38).
[0641] In a further embodiment of the GLP-1 derivative, the parent
peptide is selected from the group comprising
Arg.sup.26Lys.sup.39-GLP-1(7-39),
Arg.sup.26,34Lys.sup.36,39-GLP-1(7-39),
Gly.sup.8Arg.sup.26Lys.sup.39-GLP- -1(7-39) and
Gly.sup.8Arg.sup.26,34Lys.sup.36,39GLP-1(7-39).
[0642] In a further embodiment of the GLP-1 derivative, the parent
peptide is selected from the group comprising
Arg.sup.34Lys.sup.40-GLP-1(7-40),
Arg.sup.26,34Lys.sup.36,40GLP-1(7-40),
Gly.sup.8Arg.sup.34Lys.sup.40-GLP-- 1(7-40) and
Gly.sup.8Arg.sup.26,34Lys.sup.36,40-GLP-1(7-40).
[0643] In a further embodiment of the GLP-1 derivative, the parent
peptide is:
5 Arg.sup.26-GLP-1(7-36); Arg.sup.34-GLP-1(7-36);
Arg.sup.26,34Lys.sup.36-GLP-1(7-36); Arg.sup.26-GLP-1(7-36)amide;
Arg.sup.34-GLP-1(7-36)amide; Arg.sup.26,34Lys.sup.36-GLP-1(7-36)-
amide; Arg.sup.26-GLP-1(7-37); Arg.sup.34-GLP-1(7-37);
Arg.sup.26,34Lys.sup.36-GLP-1(7-37); Arg.sup.26-GLP-1(7-38);
Arg.sup.34-GLP-1(7-38); Arg.sup.26,34Lys GLP-1(7-38);
Arg.sup.26-GLP-1(7-39); Arg.sup.34-GLP-1(7-39);
Arg.sup.26,34Lys.sup.39-G- LP-1(7-39);
Gly.sup.8Arg.sup.26-GLP-1(7-36); Gly.sup.8Arg.sup.34-GLP-1(7-36);
Gly.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7- -36);
Gly.sup.8Arg.sup.26-GLP-1(7-36)amide;
Gly.sup.8Arg.sup.34-GLP-1(7-36)amide;
Gly.sup.8Arg.sup.26,34Lys.sup.36-GL- P-1(7-36)amide;
Gly.sup.8Arg.sup.26-GLP-1(7-37); Gly.sup.8Arg.sup.34-GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7- -37);
Gly.sup.8Arg.sup.26-GLP-1(7-38); Gly.sup.8Arg.sup.34-GLP-1(7-38);
Gly.sup.8Arg.sup.26,34Lys.sup.38GLP-1(7-- 38);
Gly.sup.8Arg.sup.26-GLP-1(7-39); Gly.sup.8Arg.sup.34-GLP-1(7-39);
Gly.sup.8Arg.sup.26,34Lys.sup.39-GLP-1(7- -39);
Val.sup.8Arg.sup.26-GLP-1(7-36); Val.sup.8Arg.sup.34-GLP-1(7-36);
Val.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7- -36);
Val.sup.8Arg.sup.26-GLP-1(7-36)amide;
Val.sup.8Arg.sup.34-GLP-1(7-36)amide;
Val.sup.8Arg.sup.26,34Lys.sup.36-GL- P-1(7-36)amide;
Val.sup.8Arg.sup.26-GLP-1(7-37); Val.sup.8Arg.sup.34-GLP-1(7-37);
Val.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7- -37);
Val.sup.8Arg.sup.26-GLP-1(7-38); Val.sup.8Arg.sup.34-GLP-1(7-38);
Val.sup.8Arg.sup.26,34Lys.sup.38GLP-1(7-- 38);
Val.sup.8Arg.sup.26-GLP-1(7-39); Val.sup.8Arg.sup.34-GLP-1(7-39);
Val.sup.8Arg.sup.26,34Lys.sup.39-GLP-1(7- -39);
Ser.sup.8Arg.sup.26-GLP-1(7-36); Ser.sup.8Arg.sup.34-GLP-1(7-36);
Ser.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7- -36);
Ser.sup.8Arg.sup.26-GLP-1(7-36)amide;
Ser.sup.8Arg.sup.34-GLP-1(7-36)amide;
Ser.sup.8Arg.sup.26,34Lys.sup.36-GL- P-1(7-36)amide;
Ser.sup.8Arg.sup.26-GLP-1(7-37); Ser.sup.8Arg.sup.34-GLP-1(7-37);
Ser.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7- -37);
Ser.sup.8Arg.sup.26-GLP-1(7-38); Ser.sup.8Arg.sup.34-GLP-1(7-38);
Ser.sup.8Arg.sup.26,34Lys.sup.38GLP-1(7-- 38);
Ser.sup.8Arg.sup.26-GLP-1(7-39); Ser.sup.8Arg.sup.34-GLP-1(7-39);
Ser.sup.8Arg.sup.26,34Lys.sup.39-GLP-1(7- -39);
Thr.sup.8Arg.sup.26-GLP-1(7-36); Thr.sup.8Arg.sup.34-GLP-1(7-36);
Thr.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7- -36);
Thr.sup.8Arg.sup.26-GLP-1(7-36)amide;
Thr.sup.8Arg.sup.34-GLP-1(7-36)amide;
Thr.sup.8Arg.sup.26,34Lys.sup.36-GL- P-1(7-36)amide;
Thr.sup.8Arg.sup.26-GLP-1(7-37); Thr.sup.8Arg.sup.34-GLP-1(7-37);
Thr.sup.8Arg.sup.26,34Lys.sup.36-GLP-1(7- -37);
Thr.sup.8Arg.sup.26-GLP-1(7-38); Thr.sup.8Arg.sup.34-GLP-1(7-38);
Thr.sup.8Arg.sup.26,34Lys.sup.38GLP-1(7-- 38);
Thr.sup.8Arg.sup.26-GLP-1(7-39); Thr.sup.8Arg.sup.34-GLP-1(7-39);
Thr.sup.8Arg.sup.26,34Lys.sup.39-GLP-1(7- -39);
Val.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-3- 6);
Val.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Val.sup.8Glu.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Val.sup.8Glu.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Val.sup.8Glu.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Val.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Val.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Val.sup.8Glu.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Val.sup.8Glu.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Val.sup.8Glu.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Val.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Val.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Val.sup.8Asp.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Val.sup.8Asp.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Val.sup.8Asp.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Val.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Val.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Val.sup.8Asp.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Val.sup.8Asp.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Val.sup.8Asp.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Ser.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Ser.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Ser.sup.8Glu.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Ser.sup.8Glu.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Ser.sup.8Glu.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Ser.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Ser.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Ser.sup.8Glu.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Ser.sup.8Glu.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Ser.sup.8Glu.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Ser.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Ser.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Ser.sup.8Asp.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Ser.sup.8Asp.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Ser.sup.8Asp.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Ser.sup.8Asp.sup.35Arg.sup.26,34Lys -GLP-1(7-36);
Ser.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Ser.sup.8Asp.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Ser.sup.8Asp.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Ser.sup.8Asp.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Thr.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Thr.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Thr.sup.8Glu.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Thr.sup.8Glu.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Thr.sup.8Glu.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Thr.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Thr.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Thr.sup.8Glu.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Thr.sup.8Glu.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Thr.sup.8Glu.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Thr.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Thr.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Thr.sup.8Asp.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Thr.sup.8Asp.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Thr.sup.8Asp.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Thr.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Thr.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Thr.sup.8Asp.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Thr.sup.8Asp.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Thr.sup.8Asp.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Gly.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Gly.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Gly.sup.8Glu.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Gly.sup.8Glu.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Gly.sup.8Glu.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Gly.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Gly.sup.8Glu.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Gly.sup.8Glu.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Gly.sup.8Glu.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Gly.sup.8Glu.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Gly.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Gly.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Gly.sup.8Asp.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Gly.sup.8Asp.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Gly.sup.8Asp.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Gly.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36);
Gly.sup.8Asp.sup.35Arg.sup.26,34Lys.sup.36-GLP-1(7-36)amide;
Gly.sup.8Asp.sup.36Arg.sup.26,34Lys.sup.37GLP-1(7-37);
Gly.sup.8Asp.sup.37Arg.sup.26,34Lys.sup.38GLP-1(7-38);
Gly.sup.8Asp.sup.38Arg.sup.26,34Lys.sup.39-GLP-1(7-39);
Arg.sup.26,34Lys.sup.18-GLP-1(7-36);
Arg.sup.26,34Lys.sup.18-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.18GLP-1(7-37);
Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Gly.sup.8Asp.sup.19Arg.sup.26,34Lys.s- up.18-GLP-1(7-36);
Gly.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18-GLP-1(7-36);
Gly.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18-GLP-1(7-36)am- ide;
Gly.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18-GLP-1(7-36)amide;
Gly.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18GLP-1(7-37);
Gly.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Gly.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Arg.sup.26,34Lys.sup.23-GLP-1(7-36);
Arg.sup.26,34Lys.sup.23-GLP-1(7-36)a- mide; Arg.sup.26,34Lys
GLP-1(7-37); Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Gly.sup.8Asp.sup.24Arg.sup.26,34Lys.s- up.23-GLP-1(7-36);
Gly.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23-GLP-1(7-36);
Gly.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23-GLP-1(7-36)am- ide;
Gly.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23-GLP-1(7-36)amide;
Gly.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23GLP-1(7-37);
Gly.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Gly.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Arg.sup.26,34Lys.sup.27-GLP-1(7-36);
Arg.sup.26,34Lys.sup.27-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.27GLP-1(7-37);
Arg.sup.26,34Lys.sup.27GLP-1(7-38);
Gly.sup.8Asp.sup.28Arg.sup.26,34Lys.s- up.27-GLP-1(7-36);
Gly.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27-GLP-1(7-36);
Gly.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27-GLP-1(7-36)am- ide;
Gly.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27-GLP-1(7-36)amide;
Gly.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27GLP-1(7-37);
Gly.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27GLP-1(7-38);
Gly.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27GLP-1(7-38);
Arg.sup.26,34Lys.sup.18-GLP-1(7-36);
Arg.sup.26,34Lys.sup.18-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.18GLP-1(7-37);
Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Val.sup.8Asp.sup.19Arg.sup.26,34Lys.s- up.18-GLP-1(7-36);
Val.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18-GLP-1(7-36);
Val.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18-GLP-1(7-36)am- ide;
Val.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18-GLP-1(7-36)amide;
Val.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18GLP-1(7-37);
Val.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Val.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Arg.sup.26,34Lys.sup.23-GLP-1(7-36);
Arg.sup.26,34Lys.sup.23-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.23GLP-1(7-37);
Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Val.sup.8Asp.sup.24Arg.sup.26,34Lys.s- up.23-GLP-1(7-36);
Val.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23-GLP-1(7-36);
Val.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23-GLP-1(7-36)am- ide;
Val.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23-GLP-1(7-36)amide;
Val.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23GLP-1(7-37);
Val.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Val.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Arg.sup.26,34Lys.sup.27-GLP-1(7-36);
Arg.sup.26,34Lys.sup.27-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.27GLP-1(7-37);
Arg.sup.26,34Lys.sup.27GLP-1(7-38);
Val.sup.8Asp.sup.28Arg.sup.26,34Lys.s- up.27-GLP-1(7-36);
Val.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27-GLP-1(7-36);
Val.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27-GLP-1(7-36)am- ide;
Val.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27-GLP-1(7-36)amide;
Val.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27GLP-1(7-37);
Val.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27GLP-1(7-38);
Val.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27GLP-1(7-38);
Arg.sup.26,34Lys.sup.18-GLP-1(7-36);
Arg.sup.26,34Lys.sup.18-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.18GLP-1(7-37);
Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Ser.sup.8Asp.sup.19Arg.sup.26,34Lys.s- up.18-GLP-1(7-36);
Ser.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18-GLP-1(7-36);
Ser.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18-GLP-1(7-36)am- ide;
Ser.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18-GLP-1(7-36)amide;
Ser.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18GLP-1(7-37);
Ser.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Ser.sup.8Asp.sup.17Arg.sup.26,34Lys GLP-1(7-38);
Arg.sup.26,34Lys.sup.23-GLP-1(7-36);
Arg.sup.26,34Lys.sup.23-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.23GLP-t(7-37);
Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Ser.sup.8Asp.sup.24Arg.sup.26,34Lys.s- up.23-GLP-1(7-36);
Ser.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23-GLP-1(7-36);
Ser.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23-GLP-1(7-36)am- ide;
Ser.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23-GLP-1(7-36)amide;
Ser.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23GLP-1(7-37);
Ser.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Ser.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Arg.sup.26,34Lys.sup.27-GLP-1(7-36);
Arg.sup.26,34Lys.sup.27-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.27GLP-1(7-37);
Arg.sup.26,34Lys.sup.27GLP-1(7-38);
Ser.sup.8Asp.sup.28Arg.sup.26,34Lys.s- up.27-GLP-1(7-36);
Ser.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27-GLP-1(7-36);
Ser.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27-GLP-1(7-36)am- ide;
Ser.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27-GLP-1(7-36)amide;
Ser.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27GLP-1(7-37);
Ser.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27GLP-1(7-38);
Ser.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27GLP-1(7-38);
Arg.sup.26,34Lys.sup.18-GLP-1(7-36);
Arg.sup.26,34Lys.sup.18-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.18GLP-1(7-37);
Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Thr.sup.8Asp.sup.19Arg.sup.26,34Lys.s- up.18-GLP-1(7-36);
Thr.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18-GLP-1(7-36);
Thr.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18-GLP-1(7-36)am- ide;
Thr.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18-GLP-1(7-36)amide;
Thr.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18GLP-1(7-37);
Thr.sup.8Asp.sup.19Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Thr.sup.8Asp.sup.17Arg.sup.26,34Lys.sup.18GLP-1(7-38);
Arg.sup.26,34Lys.sup.23-GLP-1(7-36);
Arg.sup.26,34Lys.sup.23-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.23GLP-1(7-37);
Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Thr.sup.8Asp.sup.24Arg.sup.26,34Lys.s- up.23-GLP-1(7-36);
Thr.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23-GLP-1(7-36);
Thr.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23-GLP-1(7-36)am- ide;
Thr.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23-GLP-1(7-36)amide;
Thr.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23GLP-1(7-37);
Thr.sup.8Asp.sup.24Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Thr.sup.8Asp.sup.22Arg.sup.26,34Lys.sup.23GLP-1(7-38);
Arg.sup.26,34Lys.sup.27-GLP-1(7-36);
Arg.sup.26,34Lys.sup.27-GLP-1(7-36)a- mide;
Arg.sup.26,34Lys.sup.27GLP-1(7-37);
Arg.sup.26,34Lys.sup.27GLP-1(7-3 8);
Thr.sup.8Asp.sup.28Arg.sup.26,34Lys.- sup.27-GLP-1(7-36);
Thr.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27-GLP-1(7-36)- ;
Thr.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27-GLP-1(7-36)a- mide;
Thr.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27-GLP-1(7-36)amide;
Thr.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27GLP-1(7-37);
Thr.sup.8Asp.sup.28Arg.sup.26,34Lys.sup.27GLP-1(7-38);
Thr.sup.8Asp.sup.26Arg.sup.26,34Lys.sup.27GLP-1(7-38).
[0644] In a further embodiment of the GLP-1 derivative, the parent
peptide is:
6 Arg.sup.26,34Lys.sup.36-GLP-1(7-36); Arg.sup.34Lys.sup.36-GLP-1-
(7-36); Arg.sup.26Lys.sup.36-GLP-1(7-37);
Arg.sup.34Lys.sup.36-GLP-1(7-37)- ;
Arg.sup.26Lys.sup.37-GLP-1(7-37); Arg.sup.34Lys.sup.37-GLP-1(7-37);
Arg.sup.26Lys.sup.39-GLP-1(7-39); Arg.sup.34Lys.sup.39-GLP-1(7-39);
Arg.sup.26,34Lys.sup.36,39-- GLP-1(7-39);
Arg.sup.26Lys.sup.18-GLP-1(7-36); Arg.sup.34Lys.sup.18-GLP-1(7-36);
Arg.sup.26Lys.sup.18GLP-1(7-37); Arg.sup.34Lys.sup.18GLP-1(7-37);
Arg.sup.26Lys.sup.18GLP-1(7-38); Arg.sup.34Lys.sup.18GLP-1(7-38);
Arg.sup.26Lys GLP-1(7-39); Arg.sup.34Lys.sup.18GLP-1(7-- 39);
Arg.sup.26Lys.sup.23-GLP-1(7-36); Arg.sup.34Lys.sup.23-GLP-1(7-36);
Arg.sup.26Lys.sup.23GLP-1(7-37); Arg.sup.34Lys.sup.23GLP-1(7-37);
Arg.sup.26Lys.sup.23GLP-1(7-38); Arg.sup.34Lys.sup.23GLP-1(7-38);
Arg.sup.26Lys.sup.23GLP-1(7-39); Arg.sup.34Lys.sup.23GLP-1(7-39);
Arg.sup.26Lys.sup.27-GLP-1(7-36); Arg.sup.34Lys.sup.27-GLP-1(7-36);
Arg.sup.26Lys.sup.27GLP-1(7-37); Arg.sup.34Lys.sup.27GLP-1(7-37);
Arg.sup.26Lys.sup.27GLP-1(7-38); Arg.sup.34Lys.sup.27GLP-1(7-3- 8);
Arg.sup.26Lys.sup.27GLP-1(7-39); Arg.sup.34Lys.sup.27GLP-1(7-39);
Arg.sup.26,34Lys.sup.18,36-GLP-1(7-36);
Arg.sup.26,34Lys.sup.18GLP-1(7-37- );
Arg.sup.26,34Lys.sup.18,37GLP-1(7-37);
Arg.sup.26,34Lys.sup.18,38GLP-1(7-38);
Arg.sup.26,34Lys.sup.18,39GLP-1(7-- 39);
Arg.sup.26,34Lys.sup.23,36-GLP-1(7-36);
Arg.sup.26,34Lys.sup.23GLP-1(7-37);
Arg.sup.26,34Lys.sup.23,37GLP-1(7-37)- ;
Arg.sup.26,34Lys.sup.23,38GLP-1(7-38);
Arg.sup.26,34Lys.sup.23,39GLP-1(7-39);
Arg.sup.26,34Lys.sup.27,36-GLP-1(7- -36);
Arg.sup.26,34Lys.sup.27GLP-1(7-37);
Arg.sup.26,34Lys.sup.27,37GLP-1(7-37);
Arg.sup.26,34Lys.sup.27,38GLP-1(7-- 38);
Arg.sup.26,34Lys.sup.27,39GLP-1(7-39); Gly.sup.8GLP-1(7-36);
Gly.sup.8GLP-1(7-37); Gly.sup.8GLP-1(7-38); Gly.sup.8GLP-1(7-39)
Gly.sup.8Arg.sup.26Lys.sup.36-GLP-- 1(7-36);
Gly.sup.8Arg.sup.34Lys.sup.36-GLP-1(7-36);
Gly.sup.8Arg.sup.26Lys.sup.36-GLP-1(7-37);
Gly.sup.8Arg.sup.34Lys.sup.36-GLP-1(7-37);
Gly.sup.8Arg.sup.26Lys.sup.37-- GLP-1(7-37);
Gly.sup.8Arg.sup.34Lys.sup.37-GLP-1(7-37);
Gly.sup.8Arg.sup.26Lys.sup.39-GLP-1(7-39);
Gly.sup.8Arg.sup.34Lys.sup.39-- GLP-1(7-39);
Gly.sup.8Arg.sup.26,34Lys.sup.36,39-GLP-1(7-39);
Gly.sup.8Arg.sup.26Lys.sup.18-GLP-1(7-36);
Gly.sup.8Arg.sup.34Lys.su- p.18-GLP-1(7-36);
Gly.sup.8Arg.sup.26Lys.sup.18GLP-1(7-37);
Gly.sup.8Arg.sup.34Lys.sup.18GLP-1(7-37);
Gly.sup.8Arg.sup.26Lys.sup.1- 8GLP-1(7-38);
Gly.sup.8Arg.sup.34Lys.sup.18GLP-1(7-38);
Gly.sup.8Arg.sup.26Lys.sup.18GLP-1(7-39);
Gly.sup.8Arg.sup.34Lys.sup.18GL- P-1(7-39);
Gly.sup.8Arg.sup.26Lys.sup.23-GLP-1(7-36);
Gly.sup.8Arg.sup.34Lys.sup.23-GLP-1(7-36);
Gly.sup.8Arg.sup.26Lys.sup.23G- LP-1(7-37);
Gly.sup.8Arg.sup.34Lys.sup.23GLP-1(7-37);
Gly.sup.8Arg.sup.26Lys.sup.23GLP-1(7-38);
Gly.sup.8Arg.sup.34Lys.sup.23GL- P-1(7-38);
Gly.sup.8Arg.sup.26Lys.sup.23GLP-1(7-39);
Gly.sup.8Arg.sup.34Lys.sup.23GLP-1(7-39);
Gly.sup.8Arg.sup.26Lys.sup.27-GLP-1(7-36);
Gly.sup.8Arg.sup.34Lys.sup.27-- GLP-1(7-36);
Gly.sup.8Arg.sup.26Lys.sup.27GLP-1(7-37);
Gly.sup.8Arg.sup.34Lys.sup.27GLP-1(7-37);
Gly.sup.8Arg.sup.26Lys.sup.27GL- P-1(7-38);
Gly.sup.8Arg.sup.34Lys.sup.27GLP-1(7-38);
Gly.sup.8Arg.sup.26Lys.sup.27GLP-1(7-39);
Gly.sup.8Arg.sup.34Lys.sup.27GL- P-1(7-39);
Gly.sup.8Arg.sup.26,34Lys.sup.18,36-GLP-1(7-3- 6);
Gly.sup.8Arg.sup.26,34Lys.sup.18GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys- .sup.18,37GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys.sup.18- ,38GLP-1(7-38);
Gly.sup.8Arg.sup.26,34Lys.sup.18,39GLP-1(7-39);
Gly.sup.8Arg.sup.26,34Lys.sup.23,36-GLP-1(7-36);
Gly.sup.8Arg.sup.26,34Lys.sup.23GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys.su- p.23,37GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys.sup.23,38G- LP-1(7-38);
Gly.sup.8Arg.sup.26,34Lys.sup.23,39GLP-1(7-39);
Gly.sup.8Arg.sup.26,34Lys.sup.27,36-GLP-1(7-36);
Gly.sup.8Arg.sup.26,34Lys.sup.27GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys.su- p.27,37GLP-1(7-37);
Gly.sup.8Arg.sup.26,34Lys.sup.27,34G- LP-1(7-38);
Gly.sup.8Arg.sup.26,34Lys.sup.27,39GLP-1(7-39);
Val.sup.8GLP-1(7-36); Val.sup.8GLP-1(7-37); Val.sup.8GLP-1(7-38);
Val.sup.8GLP-1(7-39) Val.sup.8Arg.sup.26Lys.sup.36-GLP-- 1(7-36);
Val.sup.8Arg.sup.34Lys.sup.36-GLP-1(7-36);
Val.sup.8Arg.sup.26Lys.sup.36-GLP-1(7-37);
Val.sup.8Arg.sup.34Lys.sup.36-GLP-1(7-37);
Val.sup.8Arg.sup.26Lys.sup.37-- GLP-1(7-37);
Val.sup.8Arg.sup.34Lys.sup.37-GLP-1(7-37);
Val.sup.8Arg.sup.26Lys.sup.39-GLP-1(7-39);
Val.sup.8Arg.sup.34Lys.sup.39-- GLP-1(7-39);
Val.sup.8Arg.sup.26,34Lys.sup.36,39-GLP-1(7-39);
Val.sup.8Arg.sup.26Lys.sup.18-GLP-1(7-36);
Val.sup.8Arg.sup.34Lys.su- p.18-GLP-1(7-36);
Val.sup.8Arg.sup.26Lys.sup.18GLP-1(7-37);
Val.sup.8Arg.sup.34Lys.sup.18GLP-1(7-37);
Val.sup.8Arg.sup.26Lys.sup.1- 8GLP-1(7-38);
Val.sup.8Arg.sup.34Lys.sup.18GLP-1(7-38);
Val.sup.8Arg.sup.26Lys.sup.18GLP-1(7-39);
Val.sup.8Arg.sup.34Lys.sup.18GL- P-1(7-39);
Val.sup.8Arg.sup.26Lys.sup.23-GLP-1(7-36);
Val.sup.8Arg.sup.34Lys.sup.23-GLP-1(7-36);
Val.sup.8Arg.sup.26Lys.sup.23G- LP-1(7-37);
Val.sup.8Arg.sup.34Lys.sup.23GLP-1(7-37);
Val.sup.8Arg.sup.26Lys.sup.23GLP-1(7-38);
Val.sup.8Arg.sup.34Lys.sup.23GL- P-1(7-38);
Val.sup.8Arg.sup.26Lys.sup.23GLP-1(7-39);
Val.sup.8Arg.sup.34Lys.sup.23GLP-1(7-39);
Val.sup.8Arg.sup.26Lys.sup.27-GLP-1(7-36);
Val.sup.8Arg.sup.34Lys.sup.27-- GLP-1(7-36);
Val.sup.8Arg.sup.26Lys.sup.27GLP-1(7-37);
Val.sup.8Arg.sup.34Lys.sup.27GLP-1(7-37);
Val.sup.8Arg.sup.26Lys.sup.27GL- P-1(7-38);
Val.sup.8Arg.sup.34Lys.sup.27GLP-1(7-38);
Val.sup.8Arg.sup.26Lys.sup.27GLP-1(7-39);
Val.sup.8Arg.sup.34Lys.sup.27GL- P-1(7-39);
Val.sup.8Arg.sup.26,34Lys.sup.18,36-GLP-1(7-3- 6);
Val.sup.8Arg.sup.26,34Lys.sup.18GLP-1(7-37);
Val.sup.8Arg.sup.26,34Lys- .sup.18,37GLP-1(7-37);
Val.sup.8Arg.sup.26,34Lys.sup.18- ,38GLP-1(7-38);
Val.sup.8Arg.sup.26,34Lys.sup.18,39GLP-1(7-39);
Val.sup.8Arg.sup.26,34Lys.sup.23,36-GLP-1(7-36);
Val.sup.8Arg.sup.26,34Lys.sup.23GLP-1(7-37);
Arg.sup.26,34Lys.sup.23,37GL- P-1(7-37);
Val.sup.8Arg.sup.26,34Lys.sup.23,38GLP-1(7-38- );
Val.sup.8Arg.sup.26,34Lys.sup.23,39GLP-1(7-39);
Val.sup.8Arg.sup.26,34L- ys.sup.27,36-GLP-1(7-36);
Val.sup.8Arg.sup.26,34Lys.sup- .27GLP-1(7-37);
Val.sup.8Arg.sup.26,34Lys.sup.27,37GLP-1(7-37);
Val.sup.8Arg.sup.26,34Lys.sup.27,38GLP-1(7-38);
Val.sup.8Arg.sup.26,34Lys.sup.27,39GLP-1(7-39).
[0645] GLP-1 analogues and derivatives which can be used according
to the present invention includes those referred to in WO 99/43705
(Novo Nordisk A/S), WO 99/43706 (Novo Nordisk A/S), WO 99/43707
(Novo Nordisk A/S), WO 98/08871 (Novo Nordisk A/S), WO 99/43708
(Novo Nordisk A/S), WO 99/43341 (Novo Nordisk A/S), WO 87/06941
(The General Hospital Corporation), WO 90/11296 (The General
Hospital Corporation), WO 91/11457 (Buckley et al.), WO 98/43658
(Eli Lilly & Co.), EP 0708179-A2 (Eli Lilly & Co.), EP
0699686-A2 (Eli Lilly & Co.) which are included herein by
reference.
[0646] However, protracted acting GLP-1 derivatives, in particular
those described in WO 98/08871 are more preferred. The most
preferred GLP-1 derivatives are those in which the parent peptide
has the formula GLP-1(7-C), wherein C is 36, 37, 38, 39, 40, 41,
42, 43, 44 and 45, wherein optionally a total of up to fifteen,
preferably up to ten amino acid residues have been exchanged with
any .alpha.-amino acid residue which can be coded for by the
genetic code, said parent peptide comprising one or two lipophilic
substituents having 4 to 40 carbon atoms, preferably from 8 to 25
carbon atoms, optionally via a spacer (such as .gamma.-Glu or
.beta.-Ala). The substituents are preferably selected from acyl
groups of straight-chained or branched fatty acids.
[0647] GLP-1 analogues and derivatives that include an N-terminal
imidazole group and optionally an unbranched C.sub.6-C.sub.10 acyl
group attached to the lysine residue in position 34 are also
embodiments of the invention.
[0648] In a still further embodiment of the invention the GLP-1
agonist is selected from exendin as well as analogs, derivatives,
and fragments thereof, e.g. exendin-3 and -4.
[0649] Examples of exendin as well as analogs, derivatives, and
fragments thereof to be included within the present invention are
those disclosed in WO 9746584 and US 5424286. US 5424286 describes
a method for stimulating insulin release with exendin
polypeptide(s). The exendin polypeptides disclosed include
HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGX; wherein X=P or Y, and
HX1X2GTFITSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS; wherein X1X2=SD
(exendin-3) or GE (exendin-4)). The exendin-3 and -4 and fragments
are useful in treatment of diabetes mellitus (types I or II) and
prevention of hyperglycaemia. They normalise hyperglycaemia through
glucose-dependent, insulin-independent and insulin-dependent
mechanisms. Exendin-4 is specific for exendin receptors, i.e. it
does not interact with vasoactive intestinal peptide receptors. WO
9746584 describes truncated versions of exendin peptide(s) for
treating diabetes. The disclosed peptides increase secretion and
biosynthesis of insulin, but reduce those of glucagon. The
truncated peptides can be made more economically than full length
versions.
[0650] In a still further embodiment of the invention the GLP-1
agonist is a non-peptide.
[0651] In a further embodiment the GLP-1 agonist is a molecule,
preferably a non-peptide, which binds to a GLP-1 receptor with an
affinity constant, K.sub.D, below 1 .mu.M, preferably below 100
nM.
[0652] Any possible combination of two or more of the embodiments
described herein, is comprised within the scope of the present
invention.
[0653] The term "GLP-1" means GLP-1(7-37) or GLP-1(7-36) amide.
[0654] The term "treatment" is defined as the management and care
of a patient for the purpose of combating the disease, condition,
or disorder and includes the administration of a GLP-1 agonist to
prevent the onset of the symptoms or complications, or alleviating
the symptoms or complications, or eliminating the disease,
condition, or disorder. Treatment includes modulate, inhibit,
decrease, reduce or arrest beta cell degeneration, such as necrosis
or apoptosis of beta cells, in particular the programmed
.beta.-cell death known as apoptosis of .beta.-cells as well as
prevention of beta cell degeneration, such as necrosis or apoptosis
of beta cells, in particular prevention of apoptosis of
.beta.-cells
[0655] The term "beta cell degeneration" is intended to mean loss
of beta cell function, beta cell dysfunction, and death of beta
cells, such as necrosis or apoptosis of beta cells.
[0656] In the present context "a GLP-1 agonist" is intended to
indicate a molecule, preferably GLP-1 or an analogue or a
derivative thereof, or exendin or an analogue or a derivative
thereof, or a non-peptide, which binds to a GLP-1 receptor with an
affinity constant, K.sub.D, below 1 .mu.M, preferably below 100 nM.
Methods for identifying GLP-1 agonists are described in WO 93/19175
(Novo Nordisk A/S).
[0657] In the present context "a GLP-1 agonist" is also intended to
comprise active metabolites and prodrugs thereof, such as active
metabolites and prodrugs of GLP-1 or an analogue or a derivative
thereof, or exendin or an analogue or a derivative thereof, or a
non-peptide. A "metabolite" is an active derivative of a GLP-1
agonist produced when the GLP-1 agonist is metabolized. A "prodrug"
is a compound which is either metabolized to a GLP-1 agonist or is
metabolized to the same metabolite(s) as a GLP-1 agonist.
[0658] In the present text, the designation "an analogue" is used
to designate a peptide wherein one or more amino acid residues of
the parent peptide have been substituted by another amino acid
residue and/or wherein one or more amino acid residues of the
parent peptide have been deleted and/or wherein one or more amino
acid residues have been added to the parent peptide. Such addition
can take place either in the peptide, at the N-terminal end or at
the C-terminal end of the parent peptide, or any combination
thereof.
[0659] The term "derivative" is used in the present text to
designate a peptide in which one or more of the amino acid residues
of the parent peptide have been chemically modified, e.g. by
alkylation, acylation, ester formation or amide formation.
[0660] The term "a GLP-1 derivative" is used in the present text to
designate a derivative of GLP-1 or an analogue thereof. In the
present text, the parent peptide from which such a derivative is
formally derived is in some places referred to as the "GLP-1
moiety" of the derivative.
[0661] For a description of suitable dosage forms, dosage ranges,
pharmaceutical formulations etc. reference is made to WO 98/08871
(Novo Nordisk A/S).
[0662] The route of administration may be any route, which
effectively transports the active compound to the appropriate or
desired site of action, such as oral, nasal, pulmonary, transdermal
or parenteral.
[0663] Pharmaceutical compositions (or medicaments) containing a
GLP-1 agonist may be administered parenterally to patients in need
of such a treatment. Parenteral administration may be performed by
subcutaneous, intramuscular 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 powder or a liquid
for the administration of the GLP-1 agonist in the form of a nasal
or pulmonal spray. As a still further option, the GLP-1 agonist can
also be administered transdermally, e.g. from a patch, optionally a
iontophoretic patch, or transmucosally, e.g. bucally. As a still
further option, the GLP-1 agonist (in particular GLP-1 or an
analogue thereof) can also be administered by gene therapy, such as
by implanting a cell line transformed with a vector such that it
secretes the GLP-1 agonist. The implanted cells may be encapsulated
in semi permeable membranes, e.g. macro- or microencapsulated. The
above mentioned possible ways to administer a GLP-1 agonist are not
considered as limiting the scope of the invention.
[0664] Pharmaceutical compositions containing a GLP-1 agonist 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.
[0665] Thus, the injectable compositions of the GLP-1 agonist can
be prepared using the conventional techniques of the pharmaceutical
industry which involves dissolving and mixing the ingredients as
appropriate to give the desired end product.
[0666] According to one procedure, the GLP-1 agonist is dissolved
in an amount of water which is somewhat less than the final volume
of the composition to be prepared. An isotonic agent, a
preservative and a buffer is added as required and the pH value of
the solution is adjusted--if necessary--using an acid, e.g.
hydrochloric acid, or a base, e.g. aqueous sodium hydroxide as
needed. Finally, the volume of the solution is adjusted with water
to give the desired concentration of the ingredients.
[0667] Examples of isotonic agents are sodium chloride, mannitol
and glycerol.
[0668] Examples of preservatives are phenol, m-cresol, methyl
p-hydroxybenzoate and benzyl alcohol.
[0669] Examples of suitable buffers are sodium acetate and sodium
phosphate.
[0670] Further to the above-mentioned components, solutions
containing a GLP-1 agonist may also contain a surfactant in order
to improve the solubility and/or the stability of the GLP-1
agonist.
[0671] A composition for nasal administration of certain peptides
may, for example, be prepared as described in European Patent No.
272097 (to Novo Nordisk A/S) or in WO 93/18785.
[0672] According to one embodiment of the present invention, the
GLP-1 agonist is provided in the form of a composition suitable for
administration by injection. Such a composition can either be an
injectable solution ready for use or it can be an amount of a solid
composition, e.g. a lyophilised product, which has to be dissolved
in a solvent before it can be injected. The injectable solution
preferably contains not less than about 2 mg/ml, preferably not
less than about 5 mg/ml, more preferred not less than about 10
mg/ml of the GLP-1 agonist and, preferably, not more than about 100
mg/ml of the GLP-1 agonist.
[0673] The GLP-1 agonist can be used in the treatment of various
diseases. The particular GLP-1 agonist to be used and the optimal
dose level for any patient will depend on the disease to be treated
and on a variety of factors including the efficacy of the specific
peptide derivative employed, the age, body weight, physical
activity, and diet of the patient, on a possible combination with
other drugs, and on the severity of the case. It is recommended
that the dosage of the GLP-1 agonist be determined for each
individual patient by those skilled in the art.
EXPERIMENTAL
Example 1
[0674] The male Zucker Diabetic Fatty fa/fa (ZDF) rat is a model of
Type 2 diabetes. The rats are insulin resistant but normoglycemic
from birth and they develop diabetes from about week 7 to week 10
of age. During the transitional period, the animals go through a
state of impaired glucose tolerance. Although the animals are
hyperinsulinemic before diabetes onset and during the early stages
of diabetes, they later lose glucose-stimulated insulin secretion
and finally become almost completely insulinopenic.
[0675] We have studied the effects of Arg.sup.34,
Lys.sup.26-E-(.gamma.-Gl- u(N-.alpha.-hexadecanoyl)))-GLP-1(7-37)
therapy during a period of time when the animals would normally
progress from having impaired glucose tolerance to having overt
Type 2 diabetes. Three groups of male ZDF rats (Genetic Models Inc,
Indianapolis, Ind., USA) were studied and dosed subcutaneously
bi-daily with either vehicle (group A), 30 (group B) or 150
.mu.g/kg (group C) of Arg.sup.34,
Lys.sup.26(N.sup..epsilon.-.epsilon-
.-(.gamma.-Glu(N.sup..epsilon.-a-hexadecanoyl)))-GLP-1(7-37), n=6
per group. Animals were between 7 and 8 weeks old when dosing was
initiated, and fed glucose levels were not different between the
groups before dosing began. However, they were elevated compared to
a group of non-diabetic Sprague-Dawley rats who had fed glucose
levels significantly below the ZDF animals (6.4.+-.0.6 vs
5.8.+-.0.8, mean.+-.SD, p<0.02). This demonstrates the relative
impaired glucose tolerant state of the ZDF animals when the study
began.
[0676] Bromodeoxyuridine (BrDU) is incorporated in newly
synthesized DNA and thus will label replicating cells. Six hours
before sacrifice the rats were given an injection of 100 mg BrDU/kg
intraperitoneally. After sacrifice the pancreata were fixed in 4%
PFA, dehydrated, embedded in paraffin, and 3-4 mm sections double
stained for BrDU and insulin for the measurement of beta-cell
proliferation rate.
[0677] Insulin was stained with guinea pig anti-insulin,
peroxidase-coupled rabbit anti-guinea pig Ig, and developed with
AEC to give a red stain. BrDU was stained by monoclonal mouse
anti-BrDU, biotinylated goat anti-mouse Ig, avidin peroxidase, and
developed with DAB and CuSO.sub.4 to give a dark brown stain. BrDU
stained nuclei of cells with insulin stained cytoplasm was examined
in more than 1500 cells per section. The examination of the
sections were carried out with the origin of the sections blinded
to the observer.
[0678] The rats treated with Arg.sup.34,
Lys.sup.26(N-.epsilon.-(.gamma.-G-
lu(N-.alpha.-hexadecanoyl)))-GLP-1(7-37) showed a dose dependent
increase in the fraction of beta-cells that had incorporated BrDU
as a result of stimulated cell proliferation (FIG. 1).
[0679] Neighbor sections were stained for insulin and the
combination of glucagon-somatostatin-pancreatic polypeptide for the
measurement of the relative mass of islet beta-cells and
nonbeta-cells. The beta-cells were stained for insulin as described
above. The nonbeta-cells were stained with a mixture of monoclonal
mouse anti-glucagon+rabbit anti-somatostatin+rabbit anti-pancreatic
polypeptide, detected by biotinylated swine anti-multible Ig's,
avidin peroxidase, and developed with DAB and CuSO.sub.4 to give a
dark brown stain. The volume fractions of beta- and nonbeta-cells
were estimated by point counting stereologic techniques.
[0680] The beta-cell fraction of the total pancreas was
significantly higher in the rats given Arg.sup.34,
Lys.sup.26(N-.epsilon.-(.gamma.-Glu(-
N-.alpha.-hexadecanoyl)))-GLP-1(7-37) at 30 ng/g for 6 weeks
compared to vehicle treated rats, while there was no further
increase in rats given doses of 150 ng/g (FIG. 2). We have shown
that volume of beta-cells after treatment with Arg.sup.34,
Lys.sup.26(N-.epsilon.-(.gamma.-Glu(N-.alpha.--
hexadecanoyl)))-GLP-1(7-37) increase at a dose (30 ng/g) where
proliferation was not seen. This difference strongly indicates that
an inhibition of apoptosis facilitated by the administered GLP-1
compound has taken place.
[0681] Furthermore, specific inhibition of beta-cell apoptosis by
GLP-1 or GLP-1 analogs, that is GLP-1 agonists, can be shown in
vitro by measuring inhibition of free fatty acid (FFA), glucose,
sulfonylurea, or cytokine induced apoptosis in beta cells.
[0682] In vitro assays for characterizing the effect of GLP-1 or
analogs thereof on the prevention of beta-cell apoptosis induced by
FFA: Briefly, pancreatic islet, e.g. rat, mouse and human, isolated
and cultured as described in, e.g. Diabetologia 19, 439, 1980;
Transplantation, 68, 597, 1999; J. Mol. Med., 77, 93, 1999,
Diabetes 48, 1230, 1999, J. Bio. Chem. 274, 18686, 1999; Proc.
Natl. Acad. Sci. 95, 2498, 1999;. J. Bio. Chem, 273, 33501, 1998;
Diabetologia 42, 55, 1999, with or without 0.1-10 mM long-chain
FFAs (2:1 oleate/palmitate; Sigma) and with GLP-1 or GLP-1 analogs.
Characterization of apoptotic beta cells can be analyzed as
described below.
[0683] In vitro assays for characterizing the effect of GLP-1 or
analogs thereof on the prevention of beta-cell apoptosis induced by
glucose or sulfonylureas: Briefly, islets can be isolated and
cultured as described in J. Bio. Chem, 273, 33501, 1998, and
incubated in 0-30 mM glucose as described in. J. Bio. Chem, 273,
33501, 1998, in order to induce apoptosis. In order to prevent the
glucose induced apoptosis the islets can be co-incubated with GLP-1
or GLP-1 analogs. Alternatively apoptosis can be induced with
100-500 microM tolbutamide as described in J. Bio. Chem, 273,
33501, 1998. In order to prevent the tolbutamide induced apoptosis
the islets can be co-incubated with GLP-1 or GLP-1 analogs.
Characterization of apoptotic beta cells can be analyzed as
described below and as in. J. Bio. Chem, 273, 33501, 1998.
[0684] In vitro assays for characterizing the effect of GLP-1 or
analogs thereof on the prevention of beta-cell apoptosis induced by
cytokines: Briefly, human and rat islets can be isolated and
cultured as described in, e.g. Diabetologia 42, 55, 1999. Cytokine
induced apoptosis of rat and human beta cells can be done as
describe in Diabetologia 42, 55, 1999. In order to prevent the
cytokine induced apoptosis the islets can be co-incubated with
GLP-1 or GLP-1 analogs. Characterization of apoptotic beta cells
can be analyzed as described below and as described in Diabetologia
42, 55, 1999.
[0685] Apoptosis and inhibition thereof can be detected in the
following way: The free 3' OH strand breaks resulting from DNA
degradation which is associated with apoptosis can be detected with
the terminal deoxynucleotidyl transferase-mediated dUTP-X3' nick
end-labeling (TUNEL) technique (J Cell Biol 199: 493, 1992) or
using the following kits e.g. In Situ Cell Death Detection kit;
Boehringer Mannheim, Mannheim or ApoTag, Oncor, Gaithersburg, Md.).
Preparation of pancreatic sections or islet cultures for apoptosis
staining using the TUNEL technique is described in (Diabetologia
42: 566, 1999 and Diabetes 48: 738, 1999).
[0686] Apoptosis can also be detected by electrophoresis of the
soluble DNA fraction isolated from cultured islets by quantifying
the ladder-like appearance as described in (PNAS 95: 2498,
1998).
[0687] Finally apoptosis can be detected by double staining of
cultured beta cells/islets with the DNA binding dyes Hoechst 33342
and propidium iodide as described in (Diabetologia 42: 55,
1999).
Example 2
[0688] Neonatal rats were sacrificed and pancreata were aseptically
isolated. Islets of Langerhans were isolated by standard
techniques. Intact islets were cultured in 6-well tissue culture
plates at 750 islets/well (Nunc, Roskilde, Denmark) with or without
40 U/ml recombinant rat interleukin 1, 100 U/ml interferon-.gamma.,
and 100 U/ml tumor necrosis factor-.alpha. (all from Peprotech,
London, UK) and the indicated doses of the GLP-1 derivative,
Arg.sup.34,
Lys.sup.26(N-.epsilon.-(.gamma.-Glu(N-.alpha.-hexadecanoyl)))-GLP-1(7-37)-
. After 24 hours of in vitro culture, single cell suspensions were
obtained from the islets by gentle triturating in trypsin. After 2
rounds of washing, islet cells were reacted with annexin V-FITC and
propedium iodide (PI) using a commercial staining kit according to
the manufactures recommendations (Pharmingen, San Diego, Calif.,
USA). At the end of incubation the samples were analyzed by flow
cytometry using a FACScalibur (Becton Dickinson, Mountain View,
Calif., USA). Dead cells were excluded by gating on PI-negative
cells, 25000 viable cells were acquired per sample. Data were
analyzed using the CellQuest software (Becton Dickinson).
[0689] FIG. 3 shows that 24 hours incubation with the cytokine mix
induced a substantial apoptosis in the neonatal rat islets cells.
When islet cells were cultures in the presence of 100 nM of
Arg.sup.34,
Lys.sup.26(N-.epsilon.-(.gamma.-Glu(N-.alpha.-hexadecanoyl)))-GLP-1(7-37)-
, an approximately 50% reduction in the cytokine-induced apoptosis
was observed. The figure also shows that Arg.sup.34,
Lys.sup.26(N-.epsilon.-(-
.gamma.-Glu(N-.alpha.-hexadecanoyl)))-GLP-1(7-37) itself had no
effect on the level of islet cell apoptosis. These data demonstrate
that Arg.sup.34,
Lys.sup.26(N-.epsilon.-(.gamma.-Glu(N-.alpha.-hexadecanoyl)))-
-GLP-1(7-37), is able to block cytokine-induced apoptosis in islet
cells ex vivo.
* * * * *