GLP-1 DERIVATIVES WITH HELIX-CONTENT EXCEEDING 25 %, FORMING PARTIALLY STRUCTURED MICELLAR-LIKE AGGREGATES

Abstract: The present invention relates to a pharmaceutical composition comprising a GLP-1 derivative of improved solubility and/or stability, and to a method for improving the solubility and/or stability of GLP-1 or a fragment or an analogue thereof.

Description: GLP-1 DERIVATIVES WITH HELIX-CONTENT EXCEEDING 25 %, FORMING PARTIALLY STRUCTURED MICEL- LAR-LIKE AGGREGATES Field of the invention The present invention relates to a pharmaceutical composition comprising a GLP-1 derivative of improved solubility and/or stability, and to a method for improving the solubility and/or stability of GLP-1 or a fragment or an analogue thereof. Background of the invention Peptides are widely used in medical practice, and since they can be produced by recombinant DNA technology it can be expected that their importance will increase also in the years to come. The hormones regulating insulin secretion belong to the so-called enteroinsuiar axis, desig- nating a group of hormones, released from the gastrointestinal mucosa in response to the presence and absorption of nutrients in the gut, which promote an early and potentiated release of insulin. The enhancing effect on insulin secretion, the so-called incretin effect, is probably essential for a normal glucose tolerance. Many of the gastrointestinal hormones, including gastrin and secretin (cholecystokinin is not insulinotropic in man), are insulinotro- pic, but the only physiologically important ones, those that are responsible for the incretin effect, are the glucose-dependent insulinotropic polypeptide, GIP, and glucagon-like peptide- 1 (GLP-1). Because of its insulinotropic effect, GIP, isolated in 1973 (1) immediately attracted considerable interest among diabetologists. However, numerous investigations carried out during the following years clearly indicated that a defective secretion of GIP was not in- voived in the pathogenesis of insulin dependent diabetes mellitus (IDDM) or non insulin- dependent diabetes mellitus (NIDDM) (2). Furthermore, as an insulinotropic hormone, GIP was found to be almost ineffective in NIDDM (2). The other incretin hormone, GLP-1 is the most potent insulinotropic substance known (3). Unlike GIP, it is surprisingly effective in stimulating insulin secretion in NIDDM patients. In addition, and in contrast to the other insuli- notropic hormones (perhaps with the exception of secretin) it also potently inhibits glucagon secretion. Because of these actions it has pronounced blood glucose lowering effects particularly in patients with NIDDM. GLP-1 , a product of the proglucagon (4), is one of the youngest members of the secretin-VIP family of peptides, but is already established as an important gut hormone with regulatory function in glucose metabolism and gastrointestinal secretion and metabolism (5). The glu- cagon gene is processed differently in the pancreas and in the intestine. In the pancreas (9), the processing leads to the formation and parallel secretion of 1 ) glucagon itself, occupying positions 33-61 of proglucagon (PG); 2) an N-terminal peptide of 30 amino acids (PG (1-30)) often called glicentin-related pancreatic peptide, GRPP (10, 11); 3) a hexapeptide corre- sponding to PG (64-69); 4) and, finally, the so-called major proglucagon fragment (PG (72- 158)), in which the two glucagon-like sequences are buried (9). Glucagon seems to be the only biologically active product. In contrast, in the intestinal mucosa, it is glucagon that is buried in a larger molecule, while the two glucagon-like peptides are formed separately (8). The following products are formed and secreted in parallel: 1) glicentin, corresponding to PG (1- 69), with the glucagon sequence occupying residues Nos. 33-61 (12); 2) GLP-1(7-36)amide (PG (78-107))amide (13), not as originally believed PG (72-107)amide or 108, which is inactive). Small amounts of C-terminally glycine-extended but equally bioactive GLP-1 (7-37), (PG (78-108)) are also formed (14); 3) intervening peptide-2 (PG (111-122)amide) (15); and 4) GLP-2 (PG (126-158)) (15, 16). A fraction of glicentin is cleaved further into GRPP (PG (1-30)) and oxyntomodulin (PG (33-69)) (17, 18). Of these peptides, GLP-1 , has the most conspicuous biological activities. The amino acid sequence of GLP-1 is given i.a. by Schmidt et al. (Diabetologia 28 704-707 (1985). Although the interesting pharmacological properties of GLP-1(7-37) and analogues thereof have attracted much attention in recent years only little is known about the structure of these molecules. The secondary structure of GLP-1 in micelles has been described by Thorton et al. (Biochemistry 33 3532-3539 (1994)), but in normal solution, GLP-1 is considered a very flexible molecule. Surprisingly, we found that derivatisation of this relatively small and very flexible molecule resulted in compounds whose plasma profile were highly protrac- ted and still had retained activity (PCT application No. DK97/00340). While much attention has been focused on the pharmacological properties of acylated GLP- 1 derivatives, hitherto little is known about their physico-chemical and solution structural properties. Such knowledge is a prerequisite for rational handling during e.g. production, purifi- cation and formulation work and is eventually important for understanding of the structural basis for the protraction mechanism. GLP-1 and analogues of GLP-1 and fragments thereof are potentially useful i.a. in the treatment of type 1 and type 2 diabetes. However, solubility limitations and the low stability against the actions of endogenous diaminopeptidyl peptidase limits the usefulness of these compounds, and thus there still is a need for improvements in this field. Accordingly, it is one object 3 of the present invention to provide pharmaceutical solutions comprising GLP-1 derivatives with improved solubility and stability. References. 1. Pederson RA. Gastric Inhibitory Polypeptide. In Walsh JH, Dockray GJ (eds) Gut peptides: Biochemistry and Physiology. Raven Press, New York 1994, pp. 217259. 2. Krarup T. Immunoreactive gastric inhibitory polypeptide. Endocr Rev 1988;9:122-134. 3. ørskov C. Glucagon-like peptide-1 , a new hormone of the enteroinsular axis. Diabetologia 1992; 35:701-711. 4. Bell Gl, Sanchez-Pescador R, Laybourn PJ, Najarian RC. Exon duplication and divergen- ce in the human preproglucagon gene. Nature 1983; 304: 368-371. 5. Hoist JJ. Glucagon-like peptide-1 (GLP-1) - a newly discovered Gl hormone. Gastroen- terology 1994; 107: 1848-1855. 6. Hoist JJ. Gut glucagon, enteroglucagon, gut GLI, glicentin - current status. Gastroentero- logy 1983;84:1602-1613. 7. Hoist JJ, ørskov C. Glucagon and other proglucagon-derived peptides. In Walsh JH, Dockray GJ, eds. Gut peptides: Biochemistry and Physiology. Raven Press, New York, pp. 305-340, 1993. 8. ørskov C, Hoist JJ, Knuhtsen S, Baldissera FGA, Poulsen SS, Nielsen OV. Glucagon-like peptides GLP-1 and GLP-2, predicted products of the glucagon gene, are secreted separately from the pig small intestine, but not pancreas. Endocrinology 1986; 119:1467-1475. 9. Hoist JJ, Bersani M, Johnsen AH, Kofod H, Hartmann B, ørskov C. Proglucagon processing in porcine and human pancreas. J Biol Chem, 1994; 269: 18827-1883. 4 10. Moody AJ, Hoist JJ, Thim L, Jensen SL. Relationship of glicentin to proglucagon and glucagon in the porcine pancreas. Nature 1981 ; 289: 514-516. 11. Thim L, Moody AJ, Purification and chemical characterisation of a glicentin-related pan- creatic peptide (proglucagon fragment) from porcine pancreas. Biochim Biophys Acta 1982;703:134-141. 12. Thim L, Moody AJ. The primary structure of glicentin (proglucagon). Regul Pept 1981 ;2:139-151. 13. ørskov C, Bersani M, Johnsen AH, Højrup P, Hoist JJ. Complete sequences of glucagon-like peptide-1 (GLP-1) from human and pig small intestine. J. Biol. Chem. 1989;264:12826-12829. 14. ørskov C, Rabenhøj L, Kofod H, Wettergren A, Hoist JJ. Production and secretion of amidated and glycine-extended glucagon-like peptide-1 (GLP-1) in man. Diabetes 1991 ; 43: 535-539. 15. Buhl T, Thim L, Kofod H, ørskov C, Harling H, & Hoist JJ: Naturally occurring products of proglucagon 111-160 in the porcine and human small intestine. J. Biol. Chem. 1988;263:8621-8624. 16. ørskov C, Buhl T, Rabenhøj L, Kofod H, Hoist JJ: Carboxypeptidase-B-like processing of the C-terminus of glucagon-like peptide-2 in pig and human small intestine. FEBS letters, 1989;247:193-106. 17. Hoist JJ. Evidence that enteroglucagon (II) is identical with the C-terminal sequence (residues 33-69) of glicentin. Biochem J. 1980;187:337-343. 18. Bataille D, Tatemoto K, Gespach C, Jornvall H, Rosselin G, Mutt V. Isolation of gluca- gon-37 (bioactive enteroglucagon/oxyntomodulin) from porcine jejuno-ileum. Characterisation of the peptide. FEBS Lett 1982;146:79-86. 5 19. ørskov C, Wettergren A, Hoist JJ. The metabolic rate and the biological effects of GLP-1 7-36amide and GLP-1 7-37 in healthy volunteers are identical. Diabetes 1993;42:658-661. 20. Elliott RM, Morgan LM, Tredger JA, Deacon S, Wright J, Marks V. Glucagon-like pepti- de-1 (7-36)amide and glucose-dependent insulinotropic polypeptide secretion in response to nutrient ingestion in man: acute post-prandial and 24-h secretion patterns. J Endocrinol 1993; 138: 159-166. 21. Kolligs F, Fehmann HC, Goke R, Goke B. Reduction of the incretin effect in rats by the glucagon-like peptide-1 receptor antagonist exendin (9-39)amide. Diabetes 1995; 44: 16-19. 22. Wang Z, Wang RM, Owji AA, Smith DM, Ghatei M, Bloom SR. Glucagon-like peptide-1 is a physiological incretin in rat. J. Clin. Invest. 1995; 95: 417-421. 23. Thorens B. Expression cloning of the pancreatic b cell receptor for the gluco-incretin hormone glucagon-like peptide 1. Proc Natl Acad Sci 1992;89:8641-4645. 24. Scrocchi L, Auerbach AB, Joyner AL, Drucker DJ. Diabetes in mice with targeted disruption of the GLP-1 receptor gene. Diabetes 1996; 45: 21A. 25. Fehmann HC, Goke R, Goke B. Cell and molecular biology of the incretin hormones glucagon-like peptide-l (GLP-1) and glucose-dependent insulin releasing polypeptide (GIP). Endocrine Reviews, 1995; 16: 390-410. 26. Gromada J, Dissing S, Bokvist K, Renstrόm E, Frøkjaer-Jensen J, Wulff BS, Rorsman P. Glucagon-like peptide I increases cytoplasmic calcium in insulin-secreting bTC3-cells by enhancement of intracellular calcium mobilisation. Diabetes 1995; 44: 767-774. 27. Holz GG, Leech CA, Habener JF. Activation of a cAMP-regulated Ca + -signaling pathway in pancreatic β-cells by the insulinotropic hormone glucagon-like peptide-1. J Biol Chem, 1996; 270: 17749-17759. 6 28. Holz GG, Kϋhltreiber WM, Habener JF. Pancreatic beta-cells are rendered glucose competent by the insulinotropic hormone glucagon-like peptide-1 (7-37). Nature 1993;361 :362-365. 29. ørskov C, Hoist JJ, Nielsen OV: Effect of truncated glucagon-like peptide-1 (proglucagon 78-107 amide) on endocrine secretion from pig pancreas, antrum and stomach. Endocrinology 1988;123:2009-2013. 30. Hvidberg A, Toft Nielsen M, Hilsted J, ørskov C, Hoist JJ. Effect of glucagon-like pepti- de-1 (proglucagon 78-107amide) on hepatic glucose production in healthy man. Metabolism 1994;43:104-108. 31. Qualmann C, Nauck M, Hoist JJ, ørskov C, Creutzfeldt W. Insulinotropic actions of intravenous glucagon-like peptide-1 [7-36 amide] in the fasting state in healthy subjects. Acta Diabetologica, 1995; 32: 13-16. 32. Nauck MA, Heimesaat MM, ørskov C, Hoist JJ, Ebert R, Creutzfeldt W. Preserved incretin activity of GLP-1 (7-36amide) but not of synthetic human GIP in patients with type 2-diabetes mellitus. J Clin Invest 1993;91 :301-307. 33. Nauck MA, Kleine N, ørskov C, Hoist JJ, Willms B, Creutzfeldt W. Normalisation of fasting hyperglycaemia by exogenous GLP-1(7-36amide) in type 2-diabetic patients. Diabetologia 1993;36:741-744. 34. Creutzfeldt W, Kleine N, Willms B, ørskov C, Hoist JJ, Nauck MA. Glucagonostatic actions and reduction of fasting hyperglycaemia by exogenous glucagon-liem, pepti- de-1(7-36amide) in type I diabetic patients. Diabetes Care 1996; 19: 580-586. 35. Schjoldager BTG, Mortensen PE, Christiansen J, ørskov C, Hoist JJ. GLP-1 (glucagon-like peptide-1 ) and truncated GLP-1 , fragments of human proglucagon, inhibit gastric acid secretion in man. Dig. Dis. Sci. 1989; 35:703-708. 7 36. Wettergren A, Schjoldager B, Mortensen PE, Myhre J, Christiansen J, Hoist JJ. Truncated GLP-1 (proglucagon 72-107amide) inhibits gastric and pancreatic functions in man. Dig Dis Sci 1993;38:665-673. 37. Layer P, Hoist JJ, Grandt D, Goebell H: Heal release of glucagon-like peptide-1 (GLP-1): association with inhibition of gastric acid in humans. Dig Dis Sci 1995; 40: 1074-1082. 38. Layer P, Hoist JJ. GLP-1 : A humoral mediator of the ileal brake in humans? Digestion 1993; 54: 385-386. 39. Nauck M, Ettler R, Niedereichholz U, ørskov C, Hoist JJ, Schmiegel W. Inhibition of gastric emptying by GLP-1(7-36 amide) or (7-37): effects on postprandial glycaemia and insulin secretion. Abstract. Gut 1995; 37 (suppl. 2): A124. 40. Schick RR, vorm Walde T, Zimmermann JP, Schusdziarra V, Classen M. Glucagon-like peptide 1 - a novel brain peptide involved in feeding regulation, in Ditschuneit H, Gries FA, Hauner H, Schusdziarra V, Wechsler JG (eds.) Obesity in Europe. John Libbey & Company ltd, 1994; pp. 363-367. 41. Tang-Christensen M, Larsen PJ, Goke R, Fink-Jensen A, Jessop DS, Møller M, Sheikh S. Brain GLP-1(7-36) amide receptors play a major role in regulation of food and water intake. Am. J. Physiol., 1996, in press. 42. Turton MD, O'Shea D, Gunn I, Beak SA, Edwards CMB, Meeran K, et al. A role for glu- cagon-like peptide-1 in the regulation of feeding. Nature 1996; 379: 69-72. 43. Willms B, Werner J, Creutzfeldt W, ørskov C, Hoist JJ, Nauck M. Inhibition of gastric emptying by glucagon-like peptide-1 (7-36 amide) in patients with type-2-diabetes mellitus. Diabetologia 1994; 37, suppl.1 : A118. 44. Larsen J, Jallad N, Damsbo P. One-week continuous infusion of GLP-1 (7-37) improves glycaemic control in NIDDM. Diabetes 1996; 45, suppl. 2: 233A. 8 45. Ritzel R, ørskov C, Hoist JJ, Nauck MA. Pharmacokinetic, insulinotropic, and glucagonostatic properties of GLP-1 [7-36 amide] after subcutaneous injection in healthy volunteers. Dose-response relationships. Diabetologia 1995; 38: 720-725. 46. Deacon CF, Johnsen AH, Hoist JJ. Degradation of glucagon-like peptide-1 by human plasma in vitro yields an N-terminally truncated peptide that is a major endogenous metabolite in vivo. J Clin Endocrinol Metab 1995; 80: 952-957. 47. Deacon CF, Nauck MA, Toft-Nielsen M, Pridal L, Willms B, Hoist JJ. 1995. Both subcu- taneous and intravenously administered glucagon-like peptide-1 are rapidly degraded from the amino terminus in type II diabetic patients and in healthy subjects. Diabetes 44: 1126-1131. Summary of the invention Human GLP-1 is a 37 amino acid residue peptide originating from preprogiucagon which is synthesised i.a. in the L-cells in the distal ileum, in the pancreas and in the brain. Processing of preprogiucagon to give GLP-1 (7-36)amide, GLP-1 (7-37) and GLP-2 occurs mainly in the L- cells. A simple system is used to describe fragments and analogues of this peptide. Thus, for example, Gly 8 -GLP-1 (7-37) designates a fragment of GLP-1 formally derived from GLP-1 by deleting the amino acid residues Nos. 1 to 6 and substituting the naturally occurring amino acid residue in position 8 (Ala) by Gly. Similarly, Lys 34 (N ε -tetradecanoyl)-GLP-1 (7-37) designates GLP-1 (7-37) wherein the ε-amino group of the Lys residue in position 34 has been tetradeca- noylated. Where reference in this text is made to C-terminally extended GLP-1 analogues, the amino acid residue in position 38 is Arg unless otherwise indicated, the optional amino acid residue in position 39 is also Arg unless otherwise indicated and the optional amino acid residue in position 40 is Asp unless otherwise indicated. Also, if a C-terminally extended analogue extends to position 41, 42, 43, 44 or 45, the amino acid sequence of this extension is as in the corresponding sequence in human preprogiucagon unless otherwise indicated. PCT application No. DK97/00340 describes various GLP-1 derivatives that are found to be very protracted. Whereas GLP-1 and GLP-1 analogues are molecules to which no defined solution structure can be ascribed, we found that some of these protracted GLP-1 derivatives g may exist in a partially structured micellar-like aggregated form which is stable over a wide concentration range. Circular Dichroism (CD) can be used to show that the GLP-1 derivatives have a certain par- tially structured conformation independent of their concentration. In contrast, for normal GLP- 1(7-37) an increase in the helix content is seen with increasing concentration, from 10-15% to 30-35% (at 500 μM concentration) in parallel with peptide self-association. For the GLP-1 derivatives forming partially structured micellar-like aggregates in aqueous solution the helix content remains constant above 30% at concentrations of 10 μM. The aggregated structured con- formation is an inherent property of the derivative present in water or dilute aqueous buffer without the need for any additional structure-inducing components. Thus, in its broadest aspect, the present invention relates to a pharmaceutical composition comprising a GLP-1 derivative which has a helix content as measured by CD at 222 nm in H 2 0 at 22 ± 2 °C exceeding 25%, preferably in the range of 25% to 50%, at a peptide concentration of about 10 μM. The size of the partially helical, micelle-like aggregates may be estimated by size-exclusion chromatography. Similarly, the apparent (critical micelle concentrations) CMC's of the pepti- des may be estimated from the concentration dependent fluorescence in the presence of appropriate dyes (e.g. Brito, R. & Vaz, W. (1986) Anal. Biochem. 152, 250-255). That the derivatives have a partially structured micellar-like aggregate conformation in aqueous solutions makes them more soluble and stable in solution as compared to the native peptide. The increased solubility and stability can be seen by comparing the solubility after 9 days of standing for a derivative and normal GLP-1(7-37) in a pharmaceutical formulation, e.g. 5 mM phosphate buffer, pH 6.9 added 0.1 M NaCI. 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 at the N-terminal end or at the C-terminal end of the parent peptide or both. 10 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 alkylati- on, acylation, ester formation or amide formation. 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. In a preferred embodiment, the present invention relates to pharmaceutical composition according to claim 1 , wherein the concentration of GLP-1 derivative is not less than 0.5 mg/ml, preferably not less than about 5 mg/ml, more preferred not less than about 10 mg/ml and, preferably, not more than about 100 mg/ml. The pharmaceutical composition of the invention preferably comprises a GLP-1 derivative wherein at least one amino acid residue of the parent peptide has a lipophilic substituent attached. More preferred are compositions comprising a GLP-1 derivative having a lipophilie-substituent which is attached to any one of the amino acid residues in position 18-38, preferably 26-34. The pharmaceutical composition according to the invention, preferably further comprises one or more of the following substances: • a pharmaceutically acceptable vehicle or carrier; • an isotonic agent, preferably selected from the group consisting of sodium chloride, mannitol and glycerol; • a preservative, preferably selected from the group consisting of phenol, m-cresol, methyl p- hydroxybenzoate, butyl p-hydroxybenzoate and benzyl alcohol; • a buffer, preferably selected from the group consisting of sodium acetate, citrate, glycylgly- cine, histidine, 2-phenylethanol and sodium phosphate; and • a surfactant capable of improving the solubility and/or the stability of the GLP-1 derivative, preferable selected from poloxymer 188, tween 20 and tween 80. 11 In a preferred embodiment, the pharmaceutical composition of the invention comprises a GLP- 1 derivative wherein the lipophilic substituent comprises from 4 to 40 carbon atoms, preferably from 8 to 25 carbon atoms. The lipophilic substituent is preferably attached to an amino acid residue in such a way that a carboxyl group of the lipophilic substituent forms an amide bond with an amino group of the amino acid residue, or, the lipophilic substituent is attached to an amino acid residue in such a way that an amino group of the lipophilic substituent forms an amide bond with a carboxyl group of the amino acid residue. In a preferred embodiment the pharmaceutical composition according to the invention compri- ses a GLP-1 derivative wherein the lipophilic substituent is attached to the parent peptide by means of a spacer. The spacer is preferably, in one embodiment, an unbranched alkane α,ω-dicarboxylic acid group having from 1 to 7 methylene groups, preferably two methylene groups, which form a bridge between an amino group of the parent peptide and an amino group of the lipophilic sub- stituent. The spacer is preferably, in another embodiment, an amino acid residue except Cys, or a di- peptide such as Gly-Lys or any unbranched alkane α,ω-aminoacid having from 1 to 7 methylene groups, preferably 2-4 methylene groups, which form a bridge between an amino group of the parent peptide and an amino group of the lipophilic substituent. In a preferred embodiment, the lipophilic substituent comprises a partially or completely hydro- genated cyclopentanophenathrene skeleton. In another preferred embodiment, the lipophilic substituent is a straight-chain or branched alkyl group. The lipophilic substituent is preferably the acyl group of a straight-chain or branched fatty acid, the acyl group more preferably being: • selected from the group comprising CH 3 (CH 2 ) n CO-, wherein n is 4 to 38, preferably CH 3 (CH 2 ) 6 CO-, CH 3 (CH 2 ) 8 CO-, CH 3 (CH 2 ) 10 CO-, CH 3 (CH 2 ) 12 CO-, CH 3 (CH 2 ) 14 CO-, CH 3 (CH 2 ) 16 CO-, CH 3 (CH 2 ) 18 CO-, CH 3 (CH 2 ) 20 CO- and CH 3 (CH 2 ) 22 CO-; or • an acyl group of a straight-chain or branched alkane α,ω-dicarboxylic acid; or 12 • selected from the group comprising HOOC(CH 2 ) m CO-, wherein m is from 4 to 38, preferably from 4 to 24, more preferred selected from the group comprising HOOC(CH 2 ) 14 CO-, HOOC(CH 2 ) 16 CO-, HOOC(CH 2 ) 18 CO-, HOOC(CH 2 ) 20 CO- and HOOC(CH 2 ) 22 CO-. In another preferred embodiment, the lipophilic substituent is a group of the formula CH 3 (CH 2 ) p ((CH 2 ) q COOH)CHNH-CO(CH 2 ) 2 CO-, wherein p and q are integers and p+q is an integer of from 8 to 33, preferably from 12 to 28. In another preferred embodiment, the lipophilic substituent is a group of the formula CH 3 (CH 2 ) r CO-NHCH(COOH)(CH 2 ) 2 CO-, wherein r is an integer of from 10 to 24. In another preferred embodiment, the lipophilic substituent is a group of the formula CH 3 (CH 2 ) s CO-NHCH((CH 2 ) 2 COOH)CO-, wherein s is an integer of from 8 to 24. In another preferred embodiment, the lipophilic substituent is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH-CO(CH 2 ) u CH 3 , wherein u is an integer of from 8 to 18. In another preferred embodiment, the lipophilic substituent is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH-COCH((CH 2 ) 2 COOH)NH-CO(CH 2 ) w CH 3 , wherein w is an integer of from 10 to 16. In another preferred embodiment, the lipophilic substituent is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH-CO(CH 2 ) 2 CH(COOH)NH-CO(CH 2 ) x CH 3 , wherein x is an integer of from 10 to 16. In another preferred embodiment, the lipophilic substituent is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH-CO(CH 2 ) 2 CH(COOH)NH-CO(CH 2 ) y CH 3 , wherein y is zero or an integer of from 1 to 22. In a preferred embodiment the pharmaceutical composition according to the invention, comprises a GLP-1 derivative wherein the parent peptide is GLP-1 (A-B) wherein A is an integer from 1 to 7 and B is an integer from 38 to 45, or an analogue thereof. The parent peptide is preferably, in one embodiment, 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); and analogues thereof. 13 The parent peptide is preferably, in another embodiment, selected from the group comprising GLP-1(1-35); GLP-1(1-36); GLP-1(1-36)amide; GLP-1(1-37); GLP-1(1-38); GLP-1(1-39); GLP- 1(1-40); GLP-1(1-41); and an analogues thereof. In yet another embodiment, the parent peptide is a GLP-1 analogue of formula I: 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 ) wherein Xaa at position 8 is Ala, Gly, Ser, Thr, Leu, lie, Val, Glu, Asp, or Lys, Xaa at position 9 is Glu, Asp, or Lys, Xaa at position 11 is Thr, Ala, Gly, Ser, Leu, He, Val, Glu, Asp, or Lys, Xaa at position 14 is Ser, Ala, Gly, Thr, Leu, lie, Val, Glu, Asp, or Lys, Xaa at position 16 is Val, Ala, Gly, Ser, Thr, Leu, He, Tyr, Glu, Asp, or Lys, Xaa at position 17 is Ser, Ala, Gly, Thr, Leu, He, Val, Glu, Asp, or Lys, Xaa at position 18 is Ser, Ala, Gly, Thr, Leu, He, Val, Glu, Asp, or Lys, Xaa at position 19 is Tyr, Phe, Trp, Glu, Asp, or Lys, Xaa at position 20 is Leu, Ala, Gly, Ser, Thr, Leu, He, Val, Glu, Asp, or Lys, Xaa at position 21 is Glu, Asp, or Lys, Xaa at position 22 is Gly, Ala, Ser, Thr, Leu, He, Val, Glu, Asp, or Lys, Xaa at position 23 is Gin, Asn, Arg, Glu, Asp, or Lys, Xaa at position 24 is Ala, Gly, Ser, Thr, Leu, He, Val, Arg, Glu, Asp, or Lys, Xaa at position 25 is Ala, Gly, Ser, Thr, Leu, He, Val, Glu, Asp, or Lys, Xaa at position 26 is Lys, Arg, Gin, Glu, Asp, or His, 14 Xaa at position 27 s Glu, Asp, or Lys, Xaa at position 30 s Ala, Gly, Ser, Thr, Leu, He, Val, Glu, Asp, or Lys, Xaa at position 31 s Trp, Phe, Tyr, Glu, Asp, or Lys, Xaa at position 32 s Leu, Gly, Ala, Ser, Thr, He, Val, Glu, Asp, or Lys, Xaa at position 33 s Val, Gly, Ala, Ser, Thr, Met, Leu, He, Glu, Asp, or Lys, Xaa at position 34 s Lys, Arg, Glu, Asp, or His, Xaa at position 35 s Gly, Ala, Ser, Thr, Leu, lie, Val, Glu, Asp, or Lys, Xaa at position 36 s Arg, Lys, Glu, Asp, or His, or Xaa at position 37 s Gly, Ala, Ser, Thr, Leu, lie, Val, Glu, Asp, or Lys, or is deleted, Xaa at position 38 s Arg, Lys, Glu, Asp, or His, or is deleted, Xaa at position 39 s Arg, Lys, Glu, Asp, or His, or is deleted, Xaa at position 40 s Asp, Glu, or Lys, or is deleted, Xaa at position 41 s Phe, Trp, Tyr, Glu, Asp, or Lys, or is deleted, Xaa at position 42 s Pro, Lys, Glu, or Asp, or is deleted, Xaa at position 43 s Glu, Asp, or Lys, or is deleted, Xaa at position 44 s Glu, Asp, or Lys, or is deleted, and Xaa at position 45 s Val, Glu, Asp, or Lys, or is deleted, or a C-1-6-ester, amide, C-1-6-alkylamide, C-1-6-dialkylamide and/or pharmaceutically accepta- ble salt thereof, provided that (i) when the amino acid at position 37, 38, 39, 40, 41 , 42, 43 or 44 is deleted, then e- ach amino acid downstream of the amino acid is also deleted, (ii) the derivative of the GLP-1 analog contains only one Lys, (iii) the ε-amino group of the Lys is substituted with a lipophilic substituent, (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. The pharmaceutical composition according to the invention preferably comprises a GLP-1 deri- vative wherein a total of up to fifteen, preferably up to ten, more preferably up to six, amino a- cid residues have been exchanged with any α-amino acid residue which can be coded for by the genetic code. The parent peptide is most preferably selected from one of the following the groups 15 Arg 26 -GLP-1(7-37); Arg 34 -GLP-1(7-37); Lys 36 -GLP- 1(7-37); Arg 2634 Lys 36 -GLP-1(7-37); Arg 2634 Lys 38 GLP-1(7-38); Arg 2634 Lys 39 -GLP-1 (7-39); Arg ^Lys^-GLP-l (7-40); Arg 26 Lys 36 - GLP-1(7-37); Arg 3 Lys 36 -GLP-1(7-37); Arg 26 Lys 39 -GLP- 1(7-39); Arg 34 Lys 0 -GLP-1 (7-40); Arg 26 ' 34 Lys 3639 -GLP-1(7-39); Arg 26 ' 34 Lys 36 0 -GLP-1(7-40); Gly 8 Arg26-GLP-1(7-37); Gly'Αrg 34 - GLP-1(7-37); Gly 8 Lys 36 -GLP-1(7-37); Gly 8 Arg 2634 Lys 35 -GLP-1(7-37); Gly 8 Arg 2634 Lys 39 -GLP- 1(7-39); Gly 8 Arg 26 ' 34 Lys 40 -GLP-1(7-40); Gly 8 Arg 26 Lys 36 -GLP- 1(7-37); Gly'Αrg^Lys -GLP-l (7- 37); Gly 8 Arg 26 Lys 39 -GLP-1(7-39); Gly 8 Arg 34 Lys 40 -GLP-1(7-40); Gly'Αrg^Lys∞∞-GLP-l (7- 39) and Gly 8 Arg 2634 Lys 36 0 -GLP-1(7-40); or Arg 263 Lys 38 GLP-1(7-38) Arg 2634 Lys 39 GLP-1 7-39); Arg 26 ' 34 Lys 40 GLP-1(7-40); Arg 2634 Lys 1 GLP-1(7-41) Arg 263 " 4 LLyyss 4422 G GLLPP--11( 7-42) Arg^Lys^GLP-K 7-43); Arg 2634 Lys 4 GLP-1(7-44) Arg 263 " 4 LLyyss 4455 G GLLPP--11(7-45) Arg^Lys∞GLP-K 1-38); Arg 2634 Lys 39 GLP-1(1-39) Arg 263 " 4 LLyyss 400 G GLLPP--11(1-40) Arg 26 ' 34 Lys 41 GLP-1( 1-41); Arg 2634 Lys 42 GLP-1(1-42) Arg 263 " 4 LLyyss 4433 G GLLPP--11(1-43) Arg ^Lys^GLP-K 1-44); Arg∞^Lys^GLP-IO^δ) Arg∞^ "LLyyss∞ 38 G GLLPP--l1(2-38) Arg 2634 Lys 39 GLP-1 ( 2-39); Arg 26 ' 34 Lys 40 GLP-1(2-40) Arg 263 " 4 LLyyss 4411 G GLLPP--11(2-41) Arg ^Lys^GLP-K 2-42); Arg 263 Lys 3 GLP-1(2-43) Arg 26 ' 3 "LLyyss 444 G GLLPP--11(2-44) Arg 2634 Lys 5 GLP-1( 2-45); Arg^Lys∞G P-l -Sδ) Arg 63 " 4 LLyyss 3399 G GLLPP--11(3-39) Arg ^Lys^GLP-K 3-40); Arg 263 Lys 41 GLP-1(3-41) Arg 263 " 4 LLyyss 422 G GLLPP--11(3-42) Arg^^Lys^GLP-K 3-43); Arg 634 Lys 4 GLP-1(3-44) Arg 263 " 4 LLyyss 4455 G GLLPP--11(3-45) Arg 2634 Lys 38 GLP-1 4-38); Arg 2634 Lys 39 GLP-1(4-39) Arg ^ "LLyyss^ 40 G GLLPP--l1( 4-40) Arg 634 Lys 41 GLP-1< '4-41); Arg 26 ' 34 Lys 42 GLP-1(4-42) Arg 26 ' 3 " 4 LLyyss 4433 G GLLPP--11( 4-43) Arg^Lys^GLP-l 4-44); Arg 2634 Lys 45 GLP-1(4-45) Arg 263 " 4 LLyyss 3388 G GLLPP--11( 5-38) Arg 263 Lys 39 GLP-1 [5-39); Arg∞^Lys^GLP-I^O) Arg 26 ' 3 " 4 LLyyss 4411 G GLLPP--11(5-41) Arg 634 Lys 42 GLP-1 .5-42); Arg 2634 Lys 43 GLP-1(5-43) Arg ^ "LLyyss^ 44 G GLLPP--l1(5-44) Arg 2634 Lys 45 GLP-1 [5-45); Arg^Lys∞GLP-l^-Sδ) Arg 263 " 4 LLyyss 3399 G GLLPP--11(6-39) Arg ^Lys^GLP-l .6-40); Arg 26 : "Lys 41 GLP-1(6-41) Arg ^ "LLyyss^ 42 G GLLPP--l1(6-42) Arg^Lys^GLP-l [6-43); Arg 2634 Lys 44 GLP-1(6-44) Arg ^Lys^GLP- 1(6-45); Arg∞Lys∞G LP-1(1-38); Arg^Lys∞GLP-IO-Sβ) Arg '^Lys '∞GLP- 1(1-36); Arg Lys∞C 3LP-1(7-38); Arg 3 Lys 38 GLP-1(7-38) Arg 26 ' 34 Lys 3638 GLF 3 -1(7-38); Arg 26 ' 3 Lys 5 38 GLP-1(7-38); Arg 26 Lys 39 GLP-1(1-39) Arg^Lys∞GLP-lf l-SΘ^ Arg^Lys 36 GLP-1(1-39); Arg 26 Lys 39 GLP-1(7-39) Arg* lys 39 G LP-1( 7-39) and Arg 2634 Ly; 3 36,39 GLp . 1 (7 . 3g) 16 The present invention furthermore relates to a method for improving the solubility and/or stability of GLP-1 or a fragment or an analogue thereof, characterised in that a lipophilic substituent is introduced on any one of the amino acid residues of the parent peptide. By this method the lipophilic substituent is preferably introduced on any one of the amino acid residues in position 1δ-3δ, preferably 26-34. The lipophilic substituent preferably comprises from 4 to 40 carbon atoms, more preferably from 8 to 25 carbon atoms. In a preferred embodiment the lipophilic substituent is the acyl group of a straight-chain or branched fatty acid; preferably selected from the group comprising CH 3 (CH 2 ) n CO-, wherein n is 4 to 38, preferably CH 3 (CH 2 ) 6 CO-, CH 3 (CH 2 ) 8 CO-, CH 3 (CH 2 ) 10 CO-, CH 3 (CH 2 ) 12 CO-, CH 3 (CH 2 ) 14 CO-, CH 3 (CH 2 ) 16 CO-, CH 3 (CH 2 ) 18 CO-, CH 3 (CH 2 ) 20 CO- and CH 3 (CH 2 ) 22 CO-. The GLP-1 parent peptide is preferably GLP-1 (A-B) wherein A is an integer from 1 to 7 and B is an integer from 38 to 45, or an analogue thereof. In one preferred embodiment, the GLP-1 is selected from the group comprising GLP-1(7-35); GLP-1(7-36); GLP-1 (7-36)amide; GLP-1(7-37); GLP-1(7-3δ); GLP-1 (7-39); GLP-1 (7-40) and GLP-1(7-41); and analogues thereof. In another preferred embodiment, the GLP-1 is selected from the group comprising GLP-1 (1- 35); GLP-1(1-36); GLP-1(1-36)amide; GLP-1(1-37); GLP-1(1-3δ); GLP-1(1-39); GLP-1(1-40); GLP-1(1-41); and an analogues thereof. Brief description of the drawing Figure 1 : Circular Dichroism (CD) at 222 nm as a function of peptide concentration for acylated GLP-1 derivatives dissolved in 10 mM tris buffer, pH δ, and 23°C. Detailed description of the invention To obtain a satisfactory protracted profile of action of the GLP-1 derivative, the lipophilic substituent attached to the GLP-1 moiety preferably comprises 4-40 carbon atoms, in particular 8- 25 carbon atoms. The lipophilic substituent may be attached to an amino group of the GLP-1 17 moiety by means of a carboxyl group of the lipophilic substituent which forms an amide bond with an amino group of the amino acid residue to which it is attached. Alternatively, the lipophilic substituent may be attached to said amino acid residue in such a way that an amino group of the lipophilic substituent forms an amide bond with a carboxyl group of the amino acid resi- due. As a further option, the lipophilic substituent may be linked to the GLP-1 moiety via an ester bond. Formally, the ester can be formed either by reaction between a carboxyl group of the GLP-1 moiety and a hydroxyl group of the substituent-to-be or by reaction between a hy- droxyl group of the GLP-1 moiety and a carboxyl group of the substituent-to-be. As a further alternative, the lipophilic substituent can be an alkyl group which is introduced into a primary amino group of the GLP-1 moiety. In one preferred embodiment of the invention, the lipophilic substituent is attached to the GLP- 1 moiety by means of a spacer in such a way that a carboxyl group of the spacer forms an amide bond with an amino group of the GLP-1 moiety. Examples of suitable spacers are suc- cinic acid, Lys, Glu or Asp, or a dipeptide such as Gly-Lys. When the spacer is succinic acid, one carboxyl group thereof may form an amide bond with an amino group of the amino acid residue, and the other carboxyl group thereof may form an amide bond with an amino group of the lipophilic substituent. 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 the- reof 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 ε-amino group of Lys and the lipophilic substituent. In one preferred embodiment, such a further spacer is succinic acid which forms an amide bond with the ε-amino group of Lys and with an amino group present in the lipophilic substituent. In another preferred embodiment such a further spa- cer is Glu or Asp which forms an amide bond with the ε-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 ε -acylated lysine residue. In another preferred embodiment of the present invention, the lipophilic substituent has a group which can be negatively charged. One preferred group which can be negatively charged is a carboxylic acid group. The parent peptide can be produced by a method which comprises culturing a host cell containing a DNA sequence encoding the polypeptide and capable of expressing the polypeptide in a 18 suitable nutrient medium under conditions permitting the expression of the peptide, after which the resulting peptide is recovered from the culture. The medium used to culture the cells may be any conventional medium suitable for growing the host cells, such as minimal or complex media containing appropriate supplements. Suitable media are available from commercial suppliers or may be prepared according to published recipes (e.g. in catalogues of the American Type Culture Collection). The peptide produced by the cells may then be recovered from the culture medium by conventional procedures including separating the host cells from the medium by centrifugation or filtration, precipitating the prote- inaceous components of the supernatant or filtrate by means of a salt, e.g. ammonium sulphate, purification by a variety of chromatographic procedures, e.g. ion exchange chromato- graphy, gel filtration chromatography, affinity chromatography, or the like, dependent on the type of peptide in question. The DNA sequence encoding the parent peptide may suitably be of genomic or cDNA origin, for instance obtained by preparing a genomic or cDNA library and screening for DNA sequences coding for all or part of the peptide by hybridisation using synthetic oligonucleotkJe probes in accordance with standard techniques (see, for example, Sambrook, J, Fritsch, EF and Ma- niatis, T, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York, 1989). The DNA sequence encoding the peptide may also be prepared synthetically by established standard methods, e.g. the phosphoamidite method described by Beaucage and Caruthers, Tetrahedron Letters 22 (1981), 1859 - 1869, or the method described by Matthes et al., EMBO Journal 3 (1984), 801 - 805. The DNA sequence may also be prepared by polyme- rase chain reaction using specific primers, for instance as described in US 4,683,202 or Saiki et al. , Science 239 (1986), 487 - 491. The DNA sequence may be inserted into any vector which may conveniently be subjected to recombinant DNA procedures, and the choice of vector will often depend on the host cell into which it is to be introduced. Thus, the vector may be an autonomously replicating vector, i.e. a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g. a plasmid. Alternatively, the vector may be one which, when introduced into a host cell, is integrated into the host cell genome and replicated together with the chromosome(s) into which it has been integrated. 19 The vector is preferably an expression vector in which the DNA sequence encoding the peptide is operably linked to additional segments required for transcription of the DNA, such as a promoter. The promoter may be any DNA sequence which shows transcriptional activity in the host cell of choice and may be derived from genes encoding proteins either homologous or heterologous to the host cell. Examples of suitable promoters for directing the transcription of the DNA encoding the peptide of the invention in a variety of host cells are well known in the art, cf. for instance Sambrook et al., supra. The DNA sequence encoding the peptide may also, if necessary, be operably connected to a suitable terminator, polyadenylation signals, transcriptional enhancer sequences, and translati- onal enhancer sequences. The recombinant vector of the invention may further comprise a DNA sequence enabling the vector to replicate in the host cell in question. The vector may also comprise a selectable marker, e.g. a gene the product of which comple- ments a defect in the host cell or one which confers resistance to a drug, e.g. ampicillin, kana- mycin, tetracyclin, chloramphenicol, neomycin, hygromycin or methotrexate. To direct a parent peptide of the present invention into the secretory pathway of the host cells, a secretory signal sequence (also known as a leader sequence, prepro sequence or pre sequ- ence) may be provided in the recombinant vector. The secretory signal sequence is joined to the DNA sequence encoding the peptide in the correct reading frame. Secretory signal sequences are commonly positioned 5' to the DNA sequence encoding the peptide. The secretory signal sequence may be that normally associated with the peptide or may be from a gene encoding another secreted protein. The procedures used to ligate the DNA sequences coding for the present peptide, the promoter and optionally the terminator and/or secretory signal sequence, respectively, and to insert them into suitable vectors containing the information necessary for replication, are well known to persons skilled in the art (cf., for instance, Sambrook et al.., supra). The host cell into which the DNA sequence or the recombinant vector is introduced may be any cell which is capable of producing the present peptide and includes bacteria, yeast, fungi and higher eukaryotic cells. Examples of suitable host cells well known and used in the art are, without limitation, E. coli, Saccharomyces cerevisiae, or mammalian BHK or CHO cell lines. 20 Examples of compounds which can be useful as GLP-1 moieties according to the present invention are described in International Patent Application No. WO 87/06941 (The General Hospital Corporation) which relates to a peptide fragment which comprises GLP-1(7-37) and functional derivatives thereof and to its use as an insulinotropic agent. Further GLP-1 analogues are described in International Patent Application No. 90/11296 (The General Hospital Corporation) which relates to peptide fragments which comprise GLP-1 (7-36) and functional derivatives thereof and have an insulinotropic activity which exceeds the insuli- notropic activity of GLP-1 (1-36) or GLP-1 (1-37) and to their use as insulinotropic agents. International Patent Application No. 91/11457 (Buckley et al..) discloses analogues of the active GLP-1 peptides 7-34, 7-35, 7-36, and 7-37 which can also be useful as GLP-1 moieties according to the present invention. Preparation and administration of the compositions Pharmaceutical compositions containing a GLP-1 derivative according to the present invention may be administered parenterally or orally 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 derivative in the form of a nasal or pul- monal spray. As a still further option, the pharmaceutical compositions containing a the GLP-1 derivatives of the invention can also be adapted to transdermal administration, e.g. from a patch, optionally a iontophoretic patch, or transmucosal, e.g. bucal, administration. Pharmaceutical compositions containing a GLP-1 derivative of the present invention may be prepared by conventional techniques, e.g. as described in Remington's Pharmaceutical Sciences, 1985 or in Remington: The Science and Practice of Pharmacy, 19 h edition, 1995. 21 Thus, the injectable compositions of the GLP-1 derivative of the invention 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. According to one procedure, the GLP-1 derivative 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 concen- tration of the ingredients. 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. According to one preferred embodiment of the present invention, the pharmaceutical compositions containing a GLP-1 derivative 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 0.5 mg/ml, preferably not less than about 5 mg/ml, more preferred not less than about 10 mg/ml of the GLP-1 derivative and, preferably, not more than about 100 mg/ml of the GLP-1 derivative. The pharmaceutical compositions containing a GLP-1 derivative of this invention can be used in the treatment of various diseases. The particular GLP-1 derivative 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 derivative of this inventi- on be determined for each individual patient by those skilled in the art. In particular, it is envisaged that the pharmaceutical compositions containing a GLP-1 derivative will be useful for the treatment of non-insulin dependent diabetes mellitus and/or for the treatment of obesity. 22 The present invention is further illustrated by the following examples which, however, are not to be construed as limiting the scope of protection. Examples Circular Dichroism (CD) at 222 nm as a function of peptide concentration for peptides dissolved in 10 mM tris buffer, pH 8, and 23°C was measured for GLP-1 (7-37) and the following eight GLP-1 derivatives suitable for pharmaceutical formulations according to the present in- vention: Derivative Position of lipophilic substituent (a) 26 (b) 26 (c) 23 (d) 34 (e) 38 (f) 26 (9) 26 (h) 18 (a) is Arg 3 Lys 26 (N ε -(γ-glutamyl(N α -hexadecanoyl)))-GLP-1 (7-37)-OH; (b) is Arg^Lys 26 (N ε -(γ-glutamyl(N α -lithochoyl))) GLP-1(7-37)-OH; (c) is Arg 2634 , Lys 23 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-37)-OH; (d) is Arg Lys 34 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1(7-37)-OH; (e) is Gly 8 ,Glu 37 ,Arg 26 ' 3 ,Lys 38 (N ε -(γ-glutamyl(N α -octadecanoyl))) GLP-1(7-38)-OH; (f) is Arg^Lys 26 (N ε -(hexadecanoyl)) GLP-1 (7-37)-OH; (g) is Arg^.Lys 26 (N ε -(γ-aminobutyroyl(N 1 '-hexadecanoyl))) GLP-1 (7-37)-OH; (h) is Arg 2634 , Lys 18 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-37)-OH. The results are presented in Figure 1. Note that the CD signal is proportional to the average content of α-helix in the peptides, i.e., a CD value of -1 corresponds to 10% α-helix content under these conditions. The figure shows that, as the concentration of unmodified GLP-1 (7-37) 23 is raised between 25 and 1000 μM, the content of α-helix increases form about 15% to about 30-35% in parallel with the formation of higher oligomers. In contrast with this concentration dependent behaviour, the figure shows that the helix content remains high and essentially independent of the concentration in the 1-200 μM range for a series of acylated glp-1 derivatives forming partially structured micelle-like aggregates under the same conditions. The following acronyms for commercially available chemicals are used: DMF N,N-Dimethylformamide. DCC N , N-Dicyclohexylcarbodiimide NMP N-Methyl-2-pyrrolidone. EDPA N-Ethyl-N,N-diisopropylamine. EGTA Ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid. GTP Guanosine 5'-triphosphate. TFA Trifluoroacetic acid. THF Tetrahydrofuran H-Glu(OH)-OBu l : L-Glutamic acid α-tert-butyl ester Cap-ONSu: Octanoic acid 2,5-dioxopyrrolidin-1-yl ester Lau-ONSu: Dodecanoic acid 2,5-dioxopyrrolidin-1-yl ester Myr-ONSu: Tetradecanoic acid 2,5-dioxopyrrolidin-1-yl ester. Pal-ONSu: Hexadecanoic acid 2,5-dioxopyrrolidin-1-yl ester. Ste-ONSu Octadecanoic acid 2,5-dioxopyrrolidin-1-yl ester. Abbreviations: PDMS: Plasma Desorption Mass Spectrometry MALDI-MS: Matrix Assisted Laser Desorption/lonisation Mass Spectrometry HPLC: High Performance Liquid Chromatography amu: atomic mass units Lit-Glu ONSuJ-OBu 1 : N°-Lithochoyl-L-glutamic acid α-t-butyl ester γ-2,5-dioxopyrrolidin-1-yl ester Cap-Glu(ONSu)-OBu l : N"-Octanoyl-L-glutamic acid α-t-butyl ester γ-2,5-dioxopyrrolidin-1- yl ester Cac-Glu(ONSu)-OBu': N α -Decanoyl-L-glutamic acid α-t-butyl ester γ-2,5-dioxopyrrolidin- 1-yl ester 24 Lau-Glu(ONSu)-OBu l : N α -Dodecanoyl-L-glutamic acid α-t-butyl ester γ-2,5- dioxopyrrolidin-1 -yl ester Myr-Glu(ONSu)-OBu t : N α -Tetradecanoyl-L-glutamic acid α-t-butyl ester γ-2,5- dioxopyrrolidin-1-yl ester Pal-Glu(ONSu)-OBu' : N α -Hexadecanoyl-(L)-glutamic acid α-t-butyl-γ-2,5- dioxopyrrolidin-1-yl diester. Ste-Glu(ONSu)-OBu' : N α -Octadecanoyl-(L)-glutamic acid α-t-butyl-γ-2,5- dioxopyrrolidin-1-yl diester Lau-β-Ala-ONSu: N β -Dodecanoyl-β-alanine 2,5-dioxopyrrolidin-1-yl ester Pal-β-Ala-ONSu: N β -Hexadecanoyl-β-alanine 2,5-dioxopyrrolidin-1-yl ester Lau-GABA-ONSu: N γ -Dodecanoyl-γ-aminobutyric acid 2,5-dioxopyrrolidin-1-yl ester Myr-GABA-ONSu: N γ -Tetradecanoyl-γ-aminobutyric acid 2,5-dioxopyrrolidin-1-yl ester Pal-GABA-ONSu: N γ -Hexadecanoyl-γ-aminobutyric acid 2,5-dioxopyrrolidin-1-yl ester Ste-GABA-ONSu: N γ -Octadecanoyl-γ-aminobutyric acid 2,5-dioxopyrrolidin-1-yl ester Pal-lsonip-ONSu: N-Hexadecanoyl-piperidine-4-carboxylic acid 2,5-dioxopyrrolidin-1- yl ester Pal-Glu(OBu , )-ONSu: N α -Hexadecanoyl-L-glutamic acid α-2,5-dioxopyrrolidin-1-yl ester γ-t-butyl ester HOOC-(CH 2 ) 6 -COONSu: ω-Carboxyheptanoic acid 2,5-dioxopyrrolidin-1-yl ester. HOOC-(CH 2 ) 10 -COONSu: ω-Carboxyundecanoic acid 2,5-dioxopyrrolidin-1-yl ester. HOOC-(CH 2 ) 12 -COONSu: ω-Carboxytridecanoic acid 2,5-dioxopyrrolidin-1-yl ester. HOOC-(CH 2 ) 14 -COONSu: ω-Carboxypentadecanoic acid 2,5-dioxopyrrolidin-1-yl ester. HOOC-(CH 2 ) 16 -COONSu: ω-Carboxyheptadecanoic acid 2,5-dioxopyrrolidin-1-yl ester. HOOC-(CH 2 ) 18 -COONSu: ω-Carboxynonadecanoic acid 2,5-dioxopyrrolidin-1-yl ester. Analytical Plasma Desorption Mass Spectrometry Sample preparation: The sample is dissolved in 0.1 % TFA/EtOH (1 :1) at a concentration of 1 μg/μl. The sample solution (5-10 μl) is placed on a nitrocellulose target (Bio-ion AB, Uppsala, Sweden) and allowed to adsorb to the target surface for 2 minutes. The target is subsequently rinsed with 2x25 μl 0.1 % TFA and spin-dried. Finally, the nitrocellulose target is placed in a target carrousel and introduced into the mass spectrometer. 25 MS analysis: PDMS analysis was carried out using a Bio-ion 20 time-of flight instrument (Bio-ion Nordic AB, Uppsala, Sweden). An acceleration voltage of 15 kV was applied and molecular ions formed by bombardment of the nitrocellulose surface with 252-Cf fission fragments were accelerated towards a stop detector. The resulting time-of-flight spectrum was calibrated into a true mass spectrum using the H + and NO + ions at m/z 1 and 30, respectively. Mass spectra were generally accumulated for 1.0x10 6 fission events corresponding to 15-20 minutes. Resulting assigned masses all correspond to isotopically averaged molecular masses. The accuracy of mass assignment is generally better than 0.1 %. MALDI-MS MALDI-TOF MS analysis was carried out using a Voyager RP instrument (PerSeptive Biosystems Inc., Framingham, MA) equipped with delayed extraction and operated in linear mode. Alpha-cyano-4-hydroxy-cinnamic acid was used as matrix, and mass assignments were based on external calibration. Further examples of compounds which can be useful as GLP-1 derivatives in the pharmaceutical composition according to the present invention are Example 1 Synthesis of Arg 2634 , Lys 36 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-36)-OH. To a mixture of Arg 2634 , Lys 36 GLP-1 (7-36)-OH (12.2 mg, 3.67 μmol), EDPA (13.3 mg, 103 μmol), NMP (1.71 ml) and water (655 μl) was added a solution of Pal-Glu(ONSu)- OBu 1 (5.94 mg, 11 μmol), prepared as described in PCT application no. PCT/DK97/00340, in NMP (14δ μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 90 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (6 mg, 61 μmol) in water (0.6 ml). A 0.5 % aqueous solution of ammonium-acetate (3δ ml) was added, and the resulting mixture eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aque- ous acetonitril (20 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (3.1 mg, 23 %) was isolated, and the product was analysed by PDMS. The m/z 26 value for the protonated molecular ion was found to be 3695 +- 3. The resulting molecular weight is thus 3694 +- 3 amu (theoretical value 3694 amu). Example 2 Synthesis of Arg 2634 , Lys 36 (N ε -(γ-glutamyl(N°-octadecanoyl))) GLP-1 (7-36)-OH. To a mixture of Arg 2634 , Lys 36 GLP-1 (7-36)-OH (12.2 mg, 3.7 μmol), EDPA (13.3 mg, 103 μmol), NMP (1.71 ml) and water (δ55 μl) was added a solution of Ste-Glu(ONSu)- OBu' (6.25 mg, 11 μmol), prepared as described in PCT application no. PCT/DK97/00340, in NMP (1 ml). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 90 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (6 mg, 31 μmol) in water (0.6 ml). A 0.5 % aqueous solution of ammonium acetate (54 ml) was added, and the resulting mixture eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (20 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (3.7 mg, 27 %) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3723 +- 3. The resulting molecular weight is thus 3722 +- 3 amu (theoretical value 3722 amu). Example 3 Synthesis of lithocholic acid 2,5-dioxopyrrolidin-1-yl ester. To a solution of lithocholic acid (5.44 g, 14.3 mmol) in a mixture of anhydrous THF (120 ml) and anhydrous acetonitril (30 ml) was added N-hydroxysuccinimide (1.78 g, 15 mmol). The mixture was cooled to 10°C, a solution of DCC (3.44 g, 16.7 mmol) in anhydrous THF (30 ml) was added drop wise, and the resulting reaction mixture stirred for 16 h at room temperature. The reaction mixture was filtered and partitioned between dichlorometha- ne (450 ml) and 10% aqueous Na 2 CO 3 (150 ml). The phases were separated, and the orga- nic phase washed with 10% aqueous Na 2 CO 3 (150 ml), water (2x150 ml), and dried (MgSO 4 ). The solvent was concentrated in vacuo. The residue was crystallised from a mixture of dichloromethane (30 ml) and n-heptane (30 ml). The precipitate was dried in a vacuum drying oven for 36 h to give the title compound (3.46 g, 51 %). 27 Example 4 Synthesis of Lit-Glu(ONSu)-OBu'. A suspension of H-Glu(OH)-OBu'(1.2δ g, 6.33 mmol), DMF (δδ ml) and EDPA (0.δ2 g, 6.33 mmol) and lithocholic acid 2,5-dioxopyrrolidin-1-yl ester, prepared as described in example 3, was stirred for 16 h at room temperature. The reaction mixture was concentrated in vacuo and the residue dissolved in ethyl acetate (40 ml). The resulting solution was washed with 5% aqueous citric acid (2x25 ml), brine (10 ml), and filtered). The solvent was concentrated in vacuo and the residue dissolved in DMF (12 ml). The resulting solution was added drop wise to a 10% aqueous solution of citric acid whereby the product precipitates. The precipitate was collected and washed with iced water, and dried in vacuo. The crude product was recrystallised from a mixture of n-heptane (40 ml) and 2-propanol (17 ml). The precipitate was dried in a vacuum drying oven for 4 h to give the free acid intermediate. To a solution of the free acid intermediate in DMF (16 ml) was added hydroxysuccinimide (0.45 g, 3.91 mmol), followed by a solution of DCC (0.73 g, 3.56 mmol) in dichloromethane (1δ ml). The resulting mixture was stirred at ambient temperature for 1δ h, and then filtered. The filtrate was concentrated in vacuo to a solid, and the residue was dissolved in dichloromethane (25 ml), and the filtration repeated, the solvent removed in vacuo to give a foam. The residue was dissolved in refluxing n-heptane (35 ml), and the product crystallised b addition of 2-propanol. The precipitate was collected, washed with cold n-heptane, dried at 35°C in vacuo to give the title compound (1.34 g, 57%). Example 5 Synthesis of Arg^.Lys 26 (N ε -(γ-glutamyl(N α -lithochoyl))) GLP-1 (7-37)-OH. To a mixture of Arg^.Lys 26 GLP-1 (7-37)-OH (41.1 mg, 12.2 μmol), EDPA (44 mg, 340 μmol), NMP (5.76 ml) and water (2.δδ ml) was added a solution of Lit-Glu^NSuJ-OBu 1 (24 mg, 37 μmol), prepared as described in example 4, in NMP (600 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 75 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (20 mg, 26δ μmol) in water (2 ml). A 0.5 % aqueous solution of ammonium ace- tate (128 ml) was added, and the resulting mixture divided into two equal portions, and each portion eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (2x25 ml), and finally liberated from the cartridge by elution with TFA (2x25 ml). The combined eluates were concentrated in vacuo, and the residue purified by column chromato-graphy using a cyanopropyl column (Zorbax 300SB-CN) and a stan- 28 dard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (5 mg, 11 %) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3872 +- 3. The resulting molecular weight is thus 3871 +- 3 amu (theoretical value 3871 amu). Example 6 Synthesis of Arg^.Lys 34 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-37)-OH To a mixture of Arg Lys 34 GLP-1 (7-37)-OH (18 mg, 5.3 μmol), EDPA (19.3 mg, 149 μmol), NMP (2.52 ml) and water (1.26 ml) was added a solution of Pal-Glu^NSuJ-OBu' (8.6 mg, 16 μmol) in NMP (215 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 90 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (8.δ mg, 117 μmol) in water (O.δδ ml). A 0.5 % aqueous solution of ammonium acetate (50 ml) was added, and the resulting mixture eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0- 100% in 60 minutes. The title compound (6 mg, 30 %) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3752 +- 3. The resulting molecular weight is thus 3751 +- 3 amu (theoretical value 3751 amu). Example 7 Synthesis of desamino-His .Arg^Lys 34 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-37)-OH. To a mixture of desamino-His^Arg∞.Lys 34 GLP-1 (7-37)-OH (14.3 mg, 4.2 μmol), EDPA (15.3 mg, 119 μmol), NMP (2 ml) and water (1 ml) was added a solution of Pal- GluζONSuJ-OBu* (6.64 mg, 12.7 μmol) in NMP (171 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 50 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (7 mg, 99 μmol) in water (700 μl). A 0.5 % aqueous solution of ammonium acetate (42 ml) was added, and the resulting mixture eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was concentrated in vacuo, and the residue 29 purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (5.6 mg, 35 %) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 373δ +- 3. The resulting molecular weight is thus 3737 +- 3 amu (theoretical value 3737 amu). Example 8 Synthesis of Gly 8 ,Arg 26 ' 34 ,Lys 38 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-3δ)-OH. To a mixture of Gly 8 , Arg 2634 , Lys 38 GLP-1 (7-38)-OH (11.δ mg, 3.4 μmol), EDPA (12.1 mg, 94 μmol), NMP (1.65 ml) and water (0.33 ml) was added a solution of Pal- Glu(ONSu)-OBu' (5.4 mg, 10 μmol) in NMP (135 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 75 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (5.5 mg, 73.7 μmol) in water (553 μl). A 0.5 % aqueous solution of ammonium acetate (36 ml) was added, and the resulting mixture eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (5 mg, 3δ %) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3895 +- 3. The resulting molecular weight is thus 3694 +- 3 amu (theoretical value 3694 amu). Example 9 Synthesis of Gly 8 ,Glu 37 ,Arg 26 ' 34 ,Lys 38 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-3δ)-OH. To a mixture of Gly 8 ,Glu 37 ,Arg 26 ' 34 ,Lys 38 GLP-1 (7-3δ)-OH (9 mg, 2.4δ μmol), EDPA (9 mg, 69.4 μmol), NMP (1.25 ml) and water (0.63 ml) was added a solution of Pal- Glu(ONSu)-OBu' (4 mg, 7.4 μmol) in NMP (100 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 105 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (4.1 mg, 54.6 μmol) in water (410 μl). A 0.5 % aqueous solution of ammonium acetate (27 ml) was added, and the resulting mixture eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobi- 30 lised compound washed with 5% aqueous acetonitril (15 ml), and finally liberated from the cartridge by elution with TFA (15 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (2.9 mg, 29 %) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3967 +- 3. The resulting molecular weight is thus 3966 +- 3 amu (theoretical value 3967 amu). Example 10 Synthesis of Gly 8 ,Glu 37 ,Arg 2634 ,Lys 38 (N ε -(γ-glutamyl(N α -octadecanoyl))) GLP-1 (7-33)-OH. To a mixture of G^GIu^Arg^Lys 38 GLP-1 (7-38)-OH (9 mg, 2.5 μmol), EDPA (9 mg, 69.4 μmol), NMP (1.25 ml) and water (0.63 ml) was added a solution of Ste-Glu(ONSu)- OBu 1 (4.2 mg, 7.4 μmol in NMP (105 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 105 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (4.1 mg, 54.6 μmol) in water (409 μl). A 0.5 % aqueous solution of ammonium acetate (27 ml) was added, and the resulting mixture eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (15 ml), and finally liberated from the cartridge by elution with TFA (15 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0- 100% in 60 minutes. The title compound (3.2 mg, 32 %) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3995 +- 3. The resulting molecular weight is thus 3994 +- 3 amu (theoretical value 3995 amu). Example 11 Synthesis of Cap-Glu(ONSu)-OBu\ To a solution of octanoic acid (5 g, 34.7 mmol) and N-hydroxysuccinimide (4 g, 34.7 mmol) in anhydrous acetonitril (10 ml) was added a solution of DCC (7.15 g, 34.7 mmol) in anhydrous dichloromethane (15 ml), and the resulting reaction mixture stirred for 16 h at room temperature. The precipitated solid was filtered off and recrystallised from a mixture of n-heptane (40 ml) and 2-propanol (2 ml). The precipitate was dried in a vacuum drying oven for 16 h to give the intermediate Cap-ONSu. A suspension of the crude ester intermediate 31 (3.9 g, 16.2 mmol), (L)-H-Glu(OH)-OBu'(3.28 g, 16.2 mmol), DMF (26δ ml) and EDPA (2.1 g, 16.2 mmol) was stirred for 64 h at room temperature. The reaction mixture was concentrated in vacuo and the residue dissolved in ethyl acetate (50 ml). The resulting solution was washed with 5% aqueous citric acid (2x25 ml). The solvent was concentrated in vacuo and the residue dissolved in DMF (36 ml). The resulting solution was added drop wise to a 10% aqueous solution of citric acid (357 ml) and extracted with ethyl acetate (200 ml), and dried (MgSO 4 ). The solvent was concentrated in vacuo to give the crude glutamic acid intermediate. To a mixture of the crude glutamic acid intermediate, N-hydroxysuccinimide (1.65 g, 16.1 mmol) and DMF (25 ml) was added a solution of DCC (3.32 g, 16.1 mmol) in dichloro- methane (15 ml). The resulting mixture was stirred at ambient temperature for 20 h. The reaction mixture was filtered and the solvent concentrated in vacuo. The residue was purified on a silica gel column (40- 63μ), eluted with a mixture of dichloromethane and acetonitril (1 :1) to give the title compound (0.63 g, 6% over all). Example 12 Synthesis of desamino-His 7 ,Arg 26 ,Lys 34 (N ε -(γ-glutamyl(N α -octanoyl))) GLP-1 (7-37)-OH. To a mixture of desamino-His 7 ,Arg 26 ,Lys 34 GLP-1 (7-37)-OH (14.3 mg, 4.2 μmol), EDPA (15.3 mg, 119 μmol), NMP (2 ml) and water (1 ml) was added a solution of Cap- Glu(ONSu)-OBu'. (6.δ mg, 12.7 μmol), prepared as described in example 11 , in NMP (135 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 2 h at room temperature. The reaction was quenched by the addition of a solution of glycine (7 mg, 93 μmol) in water (696 μl). A 0.5 % aqueous solution of ammonium acetate (42 ml) was added, and the resulting mixture eluted onto a Varian 5g C8 Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (4.1 mg, 27 %) was isolated, and the product was analysed by PDMS. The m/z value for the proto- nated molecular ion was found to be 3626 +- 3. The resulting molecular weight is thus 3625 +- 3 amu (theoretical value 3625 amu). Example 13 Synthesis of Glu 37 , Arg 2634 , Lys 38 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-38)-OH. 32 To a mixture of Glu 37 , Arg 2634 , Lys 38 GLP-1 (7-38)-OH (17.6 mg, 4.9 μmol), EDPA (17.6 mg, 136 μmol), NMP (1.23 ml) and water (2.46 ml) was added a solution of Pal- Glu(ONSu)-OBu' (7.9 mg, 14.6 μmol) in NMP (197 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 2 h at room temperature. The reaction was quenched by the addition of a solution of glycine (δ mg, 107 μmol) in water (604 μl). A 0.5 % aqueous solution of ammonium acetate (49 ml) was added, and the resulting mixture eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (5.1 mg, 26 %) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3981 +- 3. The resulting molecular weight is thus 3980 +- 3 amu (theoretical value 3961 amu). Example 14 ~ Synthesis of Arg^.Lys 26 (N ε -(γ-glutamyl(N α -octadecanoyl))) GLP-1 (7-37)-OH. To a mixture of Arg 34 GLP-1 (7-37)-OH (41.1 mg, 12.2 μmol), EDPA (44 mg, 341 μmol), NMP (5.76 ml) and water (2.δδ ml) was added a solution of Ste-Glu(ONSu)-OBu* (20.7 mg, 36.5 μmol in NMP (517 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 2 h at room temperature. The reaction was quenched by the addition of a solution of glycine (20.1 mg, 268 μmol) in water (2.01 ml). A 0.5 % aqueous solution of ammonium-acetate (120 ml) was added, and the re- suiting mixture eluted onto a Varian 5g C8 Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0- 100% in 60 minutes. The title compound (15.4 mg, 34 %) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 37δ1 +- 3. The resulting molecular weight is thus 37δ0 +- 3 amu (theoretical value 3779 amu). Example 15 33 Synthesis of Arg 3 ,Lys 26 (N ε -decanoyl) GLP-1 (7-37). To a mixture of Arg^-GLP-I (7-37)-OH (20 mg, 5.9 μmol), EDPA (21.4 mg, 165 μmol), NMP (2.δ ml) and water (1.4 ml) was added a solution of Cac-ONSu (4.δ mg, 17.7 μmol) in NMP (119 μl). The reaction mixture was gently shaken for 5 min., and then allowed to stand for an additional 2h at room temperature. The reaction was quenched by the addition of a solution of glycine (9.δ mg, 130 μmol) in water (93 μl). The resulting mixture was purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (7.4 mg, 35%) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3539.6 ± 3. The resulting molecular weight is thus 353δ.6 ± 3 amu (theoretical value 3536 amu). Example 16 Synthesis of Arg 34 ,Lys 26 (N ε -(hexadecanoyl)) GLP-1 (7-37)-OH. To a mixture of Arg 34 GLP-1 (7-37)-OH (41.1 mg, 12.2 μmol), EDPA (44 mg, 340 μmol), NMP (2.8δ ml) and water (2.δδ ml) was added a solution of Pal-ONSu (12.9 mg, 36.5 μmol) in NMP (3.3 ml). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 110 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (20.1 mg, 26δ μmol) in water (201 μl). The solvent was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (15 mg, 34 %) was isolated, and the product was analysed by PDMS. Example 17 Synthesis of Arg 2634 , Lys 27 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-37)-OH To a mixture of Arg 2634 , Lys 27 GLP-1 (7-37)-OH (11.6 mg, 3.4 μmol), EDPA (12.3 mg, 94.9 μmol), NMP (1.6 ml) and water (0.8 ml) was added a solution of Pal-Glu(ONSu)- OBu' (5.5 mg, 10.2 μmol) in NMP (137 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 90 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (5.6 mg, 74.6 μmol) in water (560 μl). A 0.5 % aqueous solution of ammonium acetate (34 ml) was added, and the resulting mixture eluted onto a Varian 5g C8 Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (15 ml), and finally liberated from the cartridge by eluti- 34 on with TFA (25 ml). The solvent was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0- 100% in 60 minutes. The title compound (2.1 mg, 16 %) was isolated, and the product was analysed by PDMS. Example 18 Synthesis of Arg 2634 , Lys 23 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-37)-OH. To a mixture of Arg 2634 , Lys 23 GLP-1 (7-37)-OH (11.6 mg, 3.4 μmol), EDPA (12.3 mg, 94.9 μmol), NMP (1.6 ml) and water (0.8 ml) was added a solution of Pal-Glu(ONSu)- OBu* (5.5 mg, 10.2 μmol) in NMP (137 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 90 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (5.6 mg, 74.6 μmol) in water (560 μl). A 0.5 % aqueous solution of ammonium acetate (34 ml) was added, and the resulting mixture eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (15 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The solvent was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0- 100% in 60 minutes. The title compound (3.1 mg, 24 %) was isolated, and the product was analysed by PDMS. Example 19 Synthesis of Arg 2634 , Lys 18 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-37)-OH To a mixture of Arg 26 ' 34 , Lys 18 GLP-1 (7-37)-OH (11.7 mg, 3.4 μmol), EDPA (12.2 mg, 94.6 μmol), NMP (1.6 ml) and water (0.6 ml) was added a solution of Pal-Glu(ONSu)- OBu* (5.5 mg, 10.2 μmol) in NMP (137 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 90 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (5.6 mg, 74.6 μmol) in water (560 μl). A 0.5 % aqueous solution of ammonium acetate (34 ml) was added, and the resulting mixture eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The solvent was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard 35 acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0- 100% in 60 minutes. The title compound (1.9 mg, 15 %) was isolated, and the product was analysed by PDMS. Example 20 Synthesis of Arg^.Lys 26 (N ε -(octanoyl)) GLP-1 (7-37)-OH. To a mixture of Arg 34 GLP-1 (7-37)-OH (41.1 mg, 12.2 μmol), EDPA (44 mg, 341 μmol), NMP (5.76 ml) and water (2.68 ml) was added a solution of Cap-ONSu (8.8 mg, 36.5 μmol, prepared as described in example 11 , in NMP (106 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 115 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (20 mg, 268 μmol) in water (200 μl). The solvent was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (18.8 mg, 44 %) was isolated, and the product was analysed by PDMS. Example 21 Synthesis of Arg^.Lys 26 (N ε -(dodecanoyl)) GLP-1 (7-37)-OH. To a mixture of Arg 34 GLP-1 (7-37)-OH (41.1 mg, 12.2 μmol), EDPA (44 mg, 341 μmol), NMP (5.76 ml) and water (2.88 ml) was added a solution of Lau-ONSu (8.8 mg, 36.5 μmol, prepared in a similar manner as described for Cap-ONSu in example 11), in NMP (271 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 100 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (20.1 mg, 268 μmol) in water (200 μl). The solvent was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (18 mg, 42 %) was isolated, and the product was analysed by PDMS. Example 22 Synthesis of Pal-GABA-ONSu. A mixture of Pal-ONSu (3 g, 8.4δ mmol), γ-aminobutyric acid (0.δ7 g, δ.4δ mmol) in DMF (200 ml) was stirred at room temperature for 60 h. The reaction mixture was filtered 36 and the filtrate was added drop wise to 10% aqueous citric acid (500 ml). The precipitated N- acylated intermediate was collected and dried in vacuo. To a suspension of the dried intermediate in DMF (35 ml) was added a solution of DCC (1.45 g, 7.0 mmol) in dichloromethane (20 ml). The resulting mixture was stirred at room temperature for 20 h, and then filtered. The solvent was removed in vacuo to give a solid residue. The residue was recrystallised from a mixture of n-heptane (50 ml) and 2-propanol (2.5 ml) to give the title compound (2.5 9, 75 %). Example 23 Synthesis of Arg 34 ,Lys 26 (N ε -(γ-aminobutyroyl(N γ -hexadecanoyl))) GLP-1 (7-37)-OH. To a mixture of Arg 34 , Lys 6 GLP-1 (7-37)-OH (41.1 mg, 12.2 μmol), EDPA (44 mg, 341 μmol), NMP (5.76 ml) and water (2.3δ ml) was added a solution of Pal-GABA-ONSu (16 mg, 36.5 μmol, prepared as described in example 22) in NMP (400 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additio- nal 100 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (20 mg, 268 μmol) in water (200 μl). The solvent was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB- CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (15.8 mg, 35 %) was isolated, and the product was analysed by PDMS. Example 24 Synthesis of N α -hexadecanoyl-D-glutamic acid α-t-butyl ester-γ-2,5-dioxopyrrolidin-1-yl ester. A mixture of Pal-ONSu (6.64 g, 18.δ mmol), D-glutamic acid α-tert-butyl ester (4.5 g, 1δ.δ mmol) and EDPA (4.65 g, 37.5 mmol) in DMF (53δ ml) was stirred at room temperature for 60 h. The solvent was removed and the residue dissolved in ethyl acetate (175 ml). The resulting solution was extracted with 10% aqueous citric acid (2x125 ml), and the organic phase concentrated in vacuo. The residue was dissolved in DMF (60 ml), and the resulting mixture slowly added to 10% aqueous citric acid (500 ml). The precipitated com- pound was collected and dried in vacuo, to give the crude N-acylated glutamic acid intermediate. The crude intermediate was dissolved in DMF (35 ml), and a solution of DCC (3.5 g, 17 mmol) in dichloromethane (70 ml) was added. The resulting mixture was stirred at room temperature for 20 h, and then filtered. The filtrate was concentrated in vacuo, and the solid 37 residue recrystallised from a mixture of n-heptane (75 ml) and 2-propanol (5 ml), to give the title compound (5.2 g, 50 %) Example 25 Synthesis of Arg 34 , Lys 26 (N ε -(γ-D-glutamyl(N°-hexadecanoyl))) GLP-1 (7-37)-OH. To a mixture of Arg 34 , Lys 26 GLP-1 (7-37)-OH (41.1 mg, 12.2 μmol), EDPA (44 mg, 341 μmol), NMP (5.76 ml) and water (2.8δ ml) was added a solution of N α -hexadecanoyl-D- glutamic acid α-t-butyl ester-γ-2,5-dioxopyrrolidin-1-yl ester (19.7 mg, 36.5 μmol) in NMP (491 μl). The reaction mixture was gently shaken for 5 min. at room temperature, and then allowed to stand for an additional 95 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (20 mg, 268 μmol) in water (2 ml). A 0.5 % aqueous solution of ammonium acetate (120 ml) was added, and the resulting mixture divided into to equal portions, and each portion eluted onto a Varian 5g C8 Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The combined eluates were concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (10.5 mg, 23 %) was isolated, and the product was analysed by PDMS. Example 26 Synthesis of Lys 34 (N ε -(γ-glutamyl(N α -tetradecanoyl))) GLP-1 (7-37). To a mixture of GLP-1 (7-37)-OH (33.6 mg, δ.9 μmol), EDPA (32.4 mg, 250 μmol), NMP (2.1 ml) and water (2.1 ml) was added a solution of Myr-Glu(ONSu)-OBu' (9.1 mg, 17.9 μmol), prepared as described in PCT application no. PCT/DK97/00340, in NMP (22δ μl). The reaction mixture was gently shaken for 5 min., and then allowed to stand for an additional δO min. at room temperature. The reaction was quenched by the addition of a solution of glycine (14.6 mg, 197 μmol) in water (1.47 ml). A 0.5% aqueous solution of ammonium acetate (100 ml) was added, and the resulting mixture divided into two equal portions, and each portion eluted onto a Varian 5g Cδ Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (2x25 ml), and finally liberated from the cartridge by elution with TFA (2x25 ml). The combined eluates were concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0- 38 100% in 60 minutes. The title compound (0.19 mg, 0.6%) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3693 ± 3. The resulting molecular weight is thus 3692 ± 3 amu (theoretical value 3695 amu). Example 27 Synthesis of Arg 26 ' 34 ,Lys 8 (N ε -(γ-glutamyl(N α -hexadecanoyl))) GLP-1 (7-37). To a mixture of Arg 2634 , Lys 8 -GLP-1 (7-37)-OH (10.3 mg, 3 μmol), EDPA (10.6 mg, δ3 μmol), NMP (1.44 ml) and water (0.72 ml) was added a solution of Pal-Glu(ONSu)-OBu' (4.8 mg, 8.9 μmol), prepared as described in PCT application no. PCT/DK97/00340, in NMP (120 μl). The reaction mixture was gently shaken for 5 min., and then allowed to stand for an additional 70 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (4.9 mg, 65.3 μmol) in water (490 μl). A 0.5% aqueous solution of ammonium acetate (30 ml) was added, and the resulting mixture eluted onto a Varian 5g C8 Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (3.2 mg, 28%) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3836 ± 3. The resulting molecular weight is thus 3835 ± 3 AMU (theoretical value 3836 AMU). Example 28 Synthesis of Lau-Glu(ONSu)-OBu'. To a solution of H-Glu-OBu 1 (3 g, 15 mmol) in DMF (344 ml) was added EDPA (2.58 ml, 15 mmol) and a solution of Lau-ONSu (4.5 g, 15 mmol), prepared in a similar manner as described for Cap-ONSu in example 11 , in DMF (74 ml). The resulting mixture was stirred at ambient temperature for 18 h, and the solvent removed in vacuo. The oily residue was partitioned between ethyl acetate (150 ml) and 5% aqueous citric acid (250 ml). The organic phase was concentrated in vacuo. The residue was dissolved in DMF (40 ml) and the solution added drop by drop to a 10% aqueous citric acid solution (350 ml). The precipitated product was collected, washed with water and dried in vacuo for 18 h to give the intermediate free acid. To solution of the free acid intermediate in DMF (25 ml) was added N- hydroxysuccinimide (1.7 g, 14.8 mmol) and a solution of N-(3-dimethylaminopropyl)-N'- 39 ethylcarbodiimide (2.5δ g, 13.5 mmol) in dichloromethane (52 ml). The resulting mixture was stirred at room temperature for 16 h, and the solvents removed in vacuo. The oily residue was partitioned between dichloromethane (80 ml) and water (80 ml). The organic phase was washed with 5% aqueous citric acid, dried (MgSO 4 ), and concentrated in vacuo to a solid. The solid residue was crystallised from a mixture of n-heptane (77 ml) and 2-propanol (50 ml), and finally recrystallised from n-heptane (76 ml) to give the title compound (2.96 g, 46%). Example 29 Synthesis of Arg 3 ,Lys 26 (N ε -(γ-glutamyl(N α -dodecanoyl))) GLP-1 (7-37). To a mixture of Arg^-GLP-I (7-37)-OH (20.6 mg, 6.1 μmol), EDPA (22 mg, 171 μmol), NMP (2.8δ ml) and water (1.44 ml) was added a solution Lau-Glu(ONSu)-OBu* (10.2 mg, 21.2 μmol), prepared as described in example 28, in NMP (255 μl). The reaction mixture was gently shaken for 5 min., and then allowed to stand for an additional 75 min. at room tempe- rature. The reaction was quenched by the addition of a solution of glycine (10 mg, 134 μmol) in water (100 μl). A 0.5% aqueous solution of ammonium acetate (61 ml) was added, and the resulting mixture eluted onto a Varian 5g C8 Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0- 100% in 60 minutes. The title compound (δ.2 mg, 36%) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3693 ± 3. The resulting molecular weight is 3692 ± 3 AMU (theoretical value 3693 AMU). Example 30 Synthesis of Lau-β-Ala-ONSu. To a solution of Lau-ONSu (4.25 g, 14.3 mmol), prepared in a similar manner to in DMF (400 ml) was added EDPA (1.64 g, 14.3 mmol) and β-alanine (1.27 g, 14.3 mmol). The re- suiting mixture was stirred at ambient temperature for 1δ h. Water (250 ml) and DMF (50 ml) were added and the solution stirred for 1 h at room temperature. The solvents were removed in vacuo to give a solid. The solid residue was dissolved in DMF (50 ml) and the solution added drop by drop to a 5% aqueous solution of citric acid (200 ml). The precipitate collected, washed with water (50 ml) and dried in vacuo to give the title compound (3.6 g, 93%). 40 Example 31 Synthesis of Pal-β-Ala-ONSu. To a solution of Pal-ONSu (4.25 g, 14.3 mmol) in DMF (400 ml) was added EDPA (1.84 g, 14.3 mmol) and β-alanine (1.27 g, 14.3 mmol). The resulting mixture was stirred at ambient temperature for 18 h. Water (250 ml) and DMF (50 ml) were added and the solution stirred for 1 h at room temperature. The solvents were removed in vacuo to give a solid. The solid residue was dissolved in DMF (50 ml) and the solution added drop by drop to a 5% aqueous solution of citric acid (200 ml). The precipitate collected, washed with water (50 ml) and dried in vacuo to give the title compound (3.6 g, 93%). Example 32 Synthesis of Myr-GABA-ONSu. To a solution of Myr-ONSu (4 g, 12.3 mmol) in DMF (350 ml) was added EDPA (1.58 g, 12.3 mmol) and γ-aminobutyric acid (1.26 g, 12.3 mmol). The resulting mixture was stirred at ambient temperature for 18 h. Water (50 ml) was added and the solution stirred for 1 h at room temperature. The solvents were removed in vacuo to give a solid. The solid residue was dissolved in DMF (75 ml) and the solution added drop by drop to a 5% aqueous solution of citric acid (250 ml). The precipitate collected, washed with water (100 ml) and dried in vacuo to give the free acid intermediate (3.65 g, 95%). To a solution of the free acid intermediate (3 g, 9.6 mmol), N-hydroxysuccinimide (1.65 g, 14.4 mmol) and N-(3-dimethylaminopropyl)-N'- ethylcarbodiimide hydrochloride (3.67 g, 19.1 mmol) in DMF (330 ml) was stirred for 18 h at room temperature, and the solvent removed in vacuo to give a solid. The solid residue was dissolved in dichloromethane (100 ml) and washed with brine (100 ml). The organic phase was dried (MgSO 4 ) and concentrated in vacuo to give a solid. The solid residue was recry- stallised from n-heptane (75 ml) to give the title compound (2.8 g, 71%). Example 33 Synthesis of Pal-β-Ala-ONSu. To a solution of Pal-ONSu (0.9 g, 2.8 mmol) in DMF (100 ml) were added N- hydroxysuccinimide (0.35 g, 3 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (0.79 g, 4.1 mmol). The resulting mixture was stirred at ambient temperature for 40h, and the solvent removed in vacuo. The solid residue was partitioned between water (50 ml) and 41 dichloromethane (50 ml). The organic phase was separated, dried (MgSO 4 ) and the solvent removed in vacuo to give the title compound (1.1 g, 94%). Example 34 Synthesis of Arg 34 ,Lys 26 (N ε -(β-alanyl(N α -hexadecanoyl))) GLP-1 (7-37). To a mixture of Arg M -GLP-1 (7-37)-OH (19.2 mg, 5.7 μmol), EDPA (20.5 mg, 159 μmol), NMP (2.7 ml) and water (1.35 ml) was added a solution Pal-β-Ala-ONSu (7.2 mg, 17 μmol), prepared as described in example 33, in NMP (181 μl). The reaction mixture was gently shaken for 5 min., and then allowed to stand for an additional 90 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (9.3 mg, 125 μmol) in water (93 μl). The reaction mixture was purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (11.6 mg, 55%) was isolated, and the product was analysed by PDMS. The m/z value for the protona- ted molecular ion was found to be 3694 ± 3. The resulting molecular weight is thus 3693 ± 3 AMU (theoretical value 3693 AMU). Example 35 Synthesis of Pal-Glu(OBu { )-ONSu. To a solution of H-Glu(OH)-OBu f (2.7 g, 11.3 mmol) and Pal-ONSu (3.98 g, 11.3 mmol) in DMF (300 ml) was added EDPA (3.2 g, 24.δ mmol). The resulting mixture was stirred at ambient temperature for 1δh, and the solvent concentrated in vacuo to give an oil. The oily residue was dissolved in DMF (60 ml) and the solution added drop by drop to a 10% aqueous solution of citric acid (300 ml) whereby a precipitate was formed. The precipitate was collected, washed with cold water (25 ml), and dried in vacuo to give free acid intermediate (4.44 g, 69%). The free acid intermediate (4 g, 9.1 mmol) was dissolved in DMF (50 ml) and N- hydroxysuccinimide (1.15 g, 10 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.6 g, 13.6 mmol) were added. The resulting mixture was stirred at room temperature for 60h, the solvent concentrated in vacuo to give the crude title compound (8.2 9) Example 36 Synthesis of Arg 34 ,Lys 26 (N ε -(α-glutamyl(N α -hexadecanoyl))) GLP-1 (7-37). 42 To a mixture of Arg^-GLP-I (7-37)-OH (25.6 mg, 7.6 μmol), EDPA (27.4 mg, 212 μmol), NMP (3.5 ml) and water (1.75 ml) was added a solution of Pal-Glu^Bu'J-ONSu (12.2 mg, 22.7 μmol), prepared as described in example 35, in NMP (305 μl). The reaction mixture was gently shaken for 5 min., and then allowed to stand for an additional 100 min. at room tem- perature. The reaction was quenched by the addition of a solution of glycine (12.5 mg, 168 μmol) in water (125 μl). A 0.5% aqueous solution of ammonium acetate (72.5 ml) was added, and the resulting mixture eluted onto a Varian 5g C8 Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (30 ml). The eluate was concentrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (6.1 mg, 22%) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3751 ± 3. The resulting molecular weight is thus 3750 ± 3 AMU (theoretical value 3751 AMU). Example 37 Synthesis of Ste-GABA-ONSu. To a solution of Ste-ONSu (3 g, 7.9 mmol) in DMF (270 ml) was added EDPA (1 g, 7.9 mmol) and a solution of γ-aminobutyric acid (0.81 g, 7.9 mmol) in water (40 ml). The resulting suspension was stirred at ambient temperature for 18 h, and then concentrated in vacuo to a final volume of 50 ml. The resulting suspension was added to a 5% aqueous solution of citric acid (500 ml) whereby a precipitate is formed. The precipitate was collected and washed with water (50 ml), and dried in vacuo for 4h to give the free acid intermediate (2.8 g, 97%). To a mixture of the free acid intermediate (2.6 g, 7 mmol), N-hydroxysuccinimide (1.21 g, 10.5 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.69 g, 14 mmol) in NMP (300 ml) was stirred for 70 h, and the solvent removed in vacuo to give a solid. The solid residue was dissolved in dichloromethane (100 ml) and washed with brine (2x100 ml). The organic phase was dried (MgSO 4 ) and concentrated in vacuo to give a solid. The solid residue was recrystallised from n-heptane (75 ml) to give the title compound (2.2 g, 67%). Example 38 Synthesis of Pal-lsonip-ONSu. 43 To a suspension of 1-hexadecanoylbenzotriazole (3 g, 8.4 mmol), prepared as described in the literature (Kreutzberger; van der Goot, Arch.Pharm., 307, 1974), in DMF (350 ml) were added EDPA (1.08 g, 8.4 mmol) and a solution of piperidine-4-carboxylic acid in water ( 20 ml). The resulting suspension was stirred at room temperature for 12d, and then concentra- ted in vacuo to an oil. The oily residue was added drop by drop to a 5% aqueous solution of citric acid (300 ml) whereby a precipitate was formed. The precipitate was collected and washed with water (50 ml), dried in vacuo for 2 h to give the free acid intermediate (3 g, 97%). To a solution of the free acid intermediate (2.8 g, 7.6 mmol), N-hydroxysuccinimide (1.31 g, 11.4 mmol) in DMF (250 ml) was added N-(3-dimethylaminopropyl)-N'- ethylcarbodiimide hydrochloride (2.92 g, 15.2 mmol). The resulting mixture was stirred at ambient temperature for 18h, and the solvent removed in vacuo to give an oil. The oily residue was dissolved in dichloromethane (100 ml), washed with brine (50 ml), dried (MgSO 4 ) and concentrated in vacuo to give the crude title compound (4.1 g, quant.). Example 39 Synthesis of Arg^.Lys^N^piperidinyl^-carbony N-hexadecanoyl))) GLP-1 (7-37). To a mixture of Arg^-GLP-I (7-37)-OH (25 mg, 7.4 μmol), EDPA (26.7 mg, 207 μmol), NMP (3.5 ml) and water (1.75 ml) was added a solution Pal-lsonip-ONSu (13.7 mg, 30 μmol), prepared as described in example 38 in NMP (343 μl). The reaction mixture was gently shaken for 5 min., and then allowed to stand for an additional 90 min. at room temperature. The reaction was quenched by the addition of a solution of glycine (12.2 mg, 163 μmol) in water (122 μl). The reaction mixture was purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (12 mg, 44%) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3734 ± 3. The resulting molecular weight is thus 3733 ± 3 AMU (theoretical value 3733 AMU). Example 40 Synthesis of Arg 3 ,Lys 26 (N ε -(γ-glutamyl(N α -decanoyl))) GLP-1 (7-37). To a mixture of Arg^-GLP-I (7-37)-OH (25 mg, 7.4 μmol), EDPA (26.7 mg, 207 μmol), NMP (3.5 ml) and water (1.75 ml) was added a solution of Cac-Glu(ONSu)-OBu' (10 mg, 22.1 μmol) in NMP (252 μl). The reaction mixture was gently shaken for 5 min., and then allowed to stand for an additional 140 min. at room temperature. The reaction was quenched by the 44 addition of a solution of glycine (12.2 mg, 162 μmol) in water (122 μl). A 0.5% aqueous solution of ammonium acetate (73 ml) was added, and the resulting mixture eluted onto a Varian 5g C8 Mega Bond Elut ® , the immobilised compound washed with 5% aqueous acetonitril (25 ml), and finally liberated from the cartridge by elution with TFA (25 ml). The eluate was con- centrated in vacuo, and the residue purified by column chromatography using a cyanopropyl column (Zorbax 300SB-CN) and a standard acetonitril/TFA system. The column was heated to 65°C and the acetonitril gradient was 0-100% in 60 minutes. The title compound (12.2 mg, 45%) was isolated, and the product was analysed by PDMS. The m/z value for the protonated molecular ion was found to be 3669.7 ± 3. The resulting molecular weight is thus 3668.7 ± 3 amu (theoretical value 3667 amu). Examples of pharmaceutical formulations In the following "8 mM phosphate buffer of pH 7.4" is intended to mean: 4 mM NaH 2 PO 4 , 2H 2 O and 4 mM Na 2 HPO 4 , 2H 2 O; pH adjusted to 7.4 (using Sodium Hydroxide and/or Hydrochloric acid). In the following "Compound 1" is intended to mean: Arg 34 ,Lys 26 (N ε -( γ -Glu(N α -tetradecanoyl))) GLP-1 (7-37). In the following "Compound 2" is intended to mean: Arg 3 ,Lys 26 (N ε -( γ -Glu(N α -hexadecanoyl))) GLP-1 (7-37). In the following "Compound 3" is intended to mean: Arg 263 ,Lys 36 (N ε -( γ -Glu(N α - hexadecanoyl))) GLP-1 (7-36). In the following "Compound 4" is intended to mean: Arg 26 ,Lys 34 (N ε -( γ -Glu(N α -hexadecanoyl))) GLP-1 (7-37). In the following "Compound 5" is intended to mean: Gly 8 ,Glu 37 ,Arg 26 ' 34 ,Lys 38 (N ε -( γ -Glu(N α - hexadecanoyl))) GLP-1 (7-38). General example A Compound 2 - 7.5 mg/ml Mannitol 34-50 mg/ml 45 Phenol 5 - 7.5 mg/ml δ mM phosphate buffer of pH 7.4 Mannitol and phenol were dissolved in the phosphate buffer preadjusted to pH 7.4. Hereaf- ter, the Compound were dissolved under slow stirring. The pH were adjusted to 7.4 using Sodium Hydroxide and/or Hydrochloric Acid. Finally, the formulation were sterilised by filtration through an appropriate filter. The following specific formulations were produced using the procedure under general exam- pie A: Example A1 Compound 1 2.0 mg/ml Mannitol 3δ mg/ml Phenol 5 mg/ml δ mM phosphate - buffer of pH 7.4 ad 100 ml Example A2 Compound 1 5 mg/ml Mannitol 36.9 mg/ml Phenol 5 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example A3 Compound 1 7.5 mg/ml Mannitol 34 mg/ml Phenol 7.5 mg/ml δ mM phosphate - buffer of pH 7.4 ad 100 ml Example A4 Compound 2 2.0 mg/ml 46 Mannitol 38 mg/ml Phenol 5 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml Example A5 Compound 2 5 mg/ml Mannitol 36.9 mg/ml Phenol 5 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml Example A6 Compound 2 7.5 mg/ml Mannitol 34 mg/ml Phenol 7.5 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml Example A7 Compound 3 2.0 mg/ml Mannitol 38 mg/ml Phenol 5 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml Example A8 Compound 3 5 mg/ml Mannitol 36.9 mg/ml Phenol 5 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example A9 47 Compound 3 7.5 mg/ml Mannitol 34 mg/ml Phenol 7.5 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml Example A10 Compound 4 2.0 mg/ml Mannitol 38 mg/ml Phenol 5 mg/ml 8 mM phosphate - buffer of pH 7.4 ad 100 ml Example A11 Compound 4 5 mg/ml Mannitol 36.9 mg/ml Phenol 5 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml Example A12 Compound 4 7.5 mg/ml Mannitol 34 mg/ml Phenol 7.5 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml General example B Compound 2 - 7.5 mg/ml Mannitol 19-25 mg/ml Benzyl Alcohol 14-18 mg/ml 8 mM phosphate buffer of pH 7.4 48 Mannitol and benzyl alcohol were dissolved in the phosphate buffer preadjusted to pH 7.4. Hereafter, the Compound were dissolved under slow stirring. The pH were adjusted to 7.4 using Sodium Hydroxide and/or Hydrochloric Acid. Finally, the formulation were sterilised by filtration through an appropriate filter. The following specific formulations were produced using the procedure under general example B: Example B1 Compound 1 2.0 mg/ml Mannitol 25 mg/ml Benzyl alcohol 14 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml Example B2 Compound 1 7.5 mg/ml Mannitol 19 mg/ml Benzyl alcohol 16 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example B3 Compound 2 2.0 mg/ml Mannitol 25 mg/ml Benzyl alcohol 14 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example B4 Compound 2 7.5 mg/ml Mannitol 19 mg/ml Benzyl alcohol 18 mg/ml 6 mM phosphate 49 buffer of pH 7.4 ad 100 ml Example B5 Compound 3 2.0 mg/ml Mannitol 25 mg/ml Benzyl alcohol 14 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml Example B6 Compound 3 7.5 mg/ml Mannitol 19 mg/ml Benzyl alcohol 18 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example B7 Compound 5 2.0 mg/ml Mannitol 25 mg/ml Benzyl alcohol 14 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example Bδ Compound 5 7.5 mg/ml Mannitol 19 mg/ml Benzyl alcohol 16 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml General example C Compound 2 - 7.5 mg/ml Mannitol 42-44 mg/ml Metacresol 2.5-4.0 mg/ml 50 δ mM phosphate buffer of pH 7.4 Mannitol and metacresol were dissolved in the phosphate buffer preadjusted to pH 7.4. Hereafter, the Compound were dissolved under slow stirring. The pH were adjusted to 7.4 using Sodium Hydroxide and/or Hydrochloric Acid. Finally, the formulation were sterilised by filtration through an appropriate filter. The following specific formulations were produced using the procedure under general example C: Example C1 Compound 1 2.0 mg/ml Mannitol 44 mg/ml Metacresol 2.5 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example C2 Compound 1 7.5 mg/ml Mannitol 42 mg/ml Metacresol 4 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example C3 Compound 2 2.0 mg/ml Mannitol 44 mg/ml Metacresol 2.5 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example C4 Compound 2 7.5 mg/ml Mannitol 42 mg/ml 51 Metacresol 4 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example C5 Compound 3 2.0 mg/ml Mannitol 44 mg/ml Metacresol 2.5 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml Example C6 Compound 3 7.5 mg/ml Mannitol 42 mg/ml Metacresol 4 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml Example C7 Compound 5 2.0 mg/ml Mannitol 44 mg/ml Metacresol 2.5 mg/ml 8 mM phosphate buffer of pH 7.4 ad 100 ml Example Cδ Compound 5 7.5 mg/ml Mannitol 42 mg/ml Metacresol 4 mg/ml δ mM phosphate buffer of pH 7.4 ad 100 ml

Claims: 52 CLAIMS 1. A pharmaceutical composition comprising a GLP-1 derivative which has a helix content as measured by CD at 222 nm in H 2 O at 22 + 2 °C exceeding 25%, preferably in the range of 25% to 50%, at a peptide concentration of about 10 μM . 2. A pharmaceutical composition according to claim 1 , wherein the concentration of GLP-1 derivative is not less than 0.5 mg/ml, preferably not less than about 5 mg/ml, more preferred not less than about 10 mg/ml and, preferably, not more than about 100 mg/ml. 3. A pharmaceutical composition according to claim 1 or 2, comprising a GLP-1 derivative wherein at least one amino acid residue of the parent peptide has a lipophilic substituent attached. 4. A pharmaceutical composition according to claim 3, comprising a GLP-1 derivative having a lipophilic substituent which is attached to any one of the amino acid residues in position 18-36, preferably 26-34. 5. A pharmaceutical composition according to any one of the preceding claims, further comprising a pharmaceutically acceptable vehicle or carrier. 6. A pharmaceutical composition according to any one of the preceding claims, further comprising an isotonic agent, preferably selected from the group consisting of sodium chloride, mannitol and glycerol. 7. A pharmaceutical composition according to any one of the preceding claims, further comprising a preservative, preferably selected from the group consisting of phenol, m-cresol, methyl p-hydroxybenzoate, butyl p-hydroxybenzoate and benzyl alcohol. 8. A pharmaceutical composition according to any one of the preceding claims, further comprising a buffer, preferably selected from the group consisting of sodium acetate, citrate, glycylglycine, histidine, 2-phenylethanol and sodium phosphate. 9. A pharmaceutical composition according to any one of the preceding claims, further comprising a surfactant capable of improving the solubility and/or the stability of the GLP-1 derivative, preferable selected from poloxymer 18δ, tween 20 and tween 80. 53 10. A pharmaceutical composition according to according to any one of the preceding claims, comprising a GLP-1 derivative wherein the lipophilic substituent comprises from 4 to 40 carbon atoms, preferably from 8 to 25 carbon atoms. 11. A pharmaceutical composition according to according to any one of the preceding claims, comprising a GLP-1 derivative wherein a lipophilic substituent is attached to an amino acid residue in such a way that a carboxyl group of the lipophilic substituent forms an amide bond with an amino group of the amino acid residue. 12. A pharmaceutical composition according to any one of the claims 1-11 , comprising a GLP- 1 derivative wherein a lipophilic substituent is attached to an amino acid residue in such a way that an amino group of the lipophilic substituent forms an amide bond with a carboxyl group of the amino acid residue. 13. A pharmaceutical composition according to any one of the preceding claims, comprising a GLP-1 derivative wherein the lipophilic substituent is attached to the parent peptide by means of a spacer. 14. A pharmaceutical composition according to claim 13, wherein the spacer is an unbranched alkane α,ω-dicarboxylic acid group having from 1 to 7 methylene groups, preferably two methylene groups, which form a bridge between an amino group of the parent peptide and an amino group of the lipophilic substituent. 15. A pharmaceutical composition according to claim 13, wherein the spacer is an amino acid residue except Cys, or a dipeptide such as Gly-Lys or any unbranched alkane α,ω- aminoacid having from 1 to 7 methylene groups, preferably 2-4 methylene groups, which form a bridge between an amino group of the parent peptide and an amino group of the lipophilic substituent. 16. A pharmaceutical composition according to any one of the preceding claims, comprising a GLP-1 derivative wherein the lipophilic substituent comprises a partially or completely hy- drogenated cyclopentanophenathrene skeleton. 17. A pharmaceutical composition according to any one of claims 3 to 15, wherein the lipophilic substituent is a straight-chain or branched alkyl group. 54 18. A pharmaceutical composition according to any one of claims 3 to 15, comprising a GLP-1 derivative wherein the lipophilic substituent is the acyl group of a straight-chain or branched fatty acid. 19. A pharmaceutical composition according to claim 18 wherein the acyl group is selected from the group comprising CH 3 (CH 2 ) n CO-, wherein n is 4 to 38, preferably CH 3 (CH 2 ) 6 CO-, CH 3 (CH 2 ) 8 CO-, CH 3 (CH 2 ) 10 CO-, CH 3 (CH 2 ) 12 CO-, CH 3 (CH 2 ) 14 CO-, CH 3 (CH 2 ) 16 CO-, CH 3 (CH 2 ) 18 CO-, CH 3 (CH 2 ) 20 CO- and CH 3 (CH 2 ) 22 CO-. 20. A pharmaceutical composition according to any one of claims 3 to 15, comprising a GLP-1 derivative wherein the lipophilic substituent is an acyl group of a straight-chain or branched alkane α,ω-dicarboxylic acid. 21. A pharmaceutical composition according to claim 20 wherein the acyl group is selected from the group comprising HOOC(CH 2 ) m CO-, wherein m is from 4 to 38, preferably from 4 to 24, more preferred selected from the group comprising HOOC(CH 2 ) 14 CO-, HOOC(CH 2 ) 16 CO-, HOOC(CH 2 ) 18 CO-, HOOC(CH 2 ) 20 CO- and HOOC(CH 2 ) 22 CO-. 22. A pharmaceutical composition according to any one of claims 3 to 15, comprising a GLP-1 derivative wherein the lipophilic substituent is a group of the formula CH 3 (CH 2 ) p ((CH 2 ) q COOH)CHNH-CO(CH 2 ) 2 CO-, wherein p and q are integers and p+q is an integer of from 8 to 33, preferably from 12 to 28. 23. A pharmaceutical composition according to any one of claims 3 to 15, comprising a GLP-1 derivative, wherein the lipophilic substituent is a group of the formula CH 3 (CH 2 ) r CO- NHCH(COOH)(CH 2 ) 2 CO-, wherein r is an integer of from 10 to 24. 24. A pharmaceutical composition according to any one of claims 3 to 15, comprising a GLP-1 derivative, wherein the lipophilic substituent is a group of the formula CH 3 (CH 2 ) s CO- NHCH((CH 2 ) 2 COOH)CO-, wherein s is an integer of from 8 to 24. 25. A pharmaceutical composition according to any one of claims 3 to 15, comprising a GLP-1 derivative, wherein the lipophilic substituent is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH-CO(CH 2 ) u CH 3 , wherein u is an integer of from 8 to 18. 26. A pharmaceutical composition according to any one of claims 3 to 15, comprising a GLP-1 derivative, wherein the lipophilic substituent is a group of the formula 55 -NHCH(COOH)(CH 2 ) 4 NH-COCH((CH 2 ) 2 COOH)NH-CO(CH 2 ) w CH 3 , wherein w is an integer of from 10 to 16. 27. A pharmaceutical composition according to any one of claims 3 to 15, comprising a GLP-1 derivative, wherein the lipophilic substituent is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH-CO(CH 2 ) 2 CH(COOH)NH-CO(CH 2 ) x CH 3 , wherein x is an integer of from 10 to 16. 28. A pharmaceutical composition according to any one of claims 3 to 15, comprising a GLP-1 derivative wherein the lipophilic substituent is a group of the formula -NHCH(COOH)(CH 2 ) 4 NH-CO(CH 2 ) 2 CH(COOH)NH-CO(CH 2 ) y CH 3 , wherein y is zero or an integer of from 1 to 22. 29. A pharmaceutical composition according to any one of the preceding claims, comprising a GLP-1 derivative wherein the parent peptide is GLP-1 (A-B) wherein A is an integer from 1 to 7 and B is an integer from 38 to 45, or an analogue thereof. 30. A pharmaceutical composition according to claim 29, wherein the parent peptide is selec- ted 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); and analogues thereof. 31. A pharmaceutical composition according to claim 29, wherein the parent peptide is selected from the group comprising GLP-1 (1-35); GLP-1(1-36); GLP-1(1-36)amide; GLP-1(1- 37); GLP-1(1-38); GLP-1(1-39); GLP-1(1-40); GLP-1(1-41); and an analogues thereof. 32. A pharmaceutical composition according to any of the preceding claims comprising a GLP- 1 derivative wherein a total of up to fifteen, preferably up to ten, more preferably up to six, amino acid residues have been exchanged with any α-amino acid residue which can be coded for by the genetic code. 33. A pharmaceutical composition according to any of the preceding claims, comprising a GLP-1 derivative wherein the parent peptide is selected from the group comprising Arg 26 - GLP-1(7-37); Arg^-GLP- 1(7-37); Lys -GLP-I (7-37); Arg 2634 Lys 36 -GLP-1(7-37); Arg 2634 Lys 38 GLP-1(7-3δ); Arg 2634 Lys 39 -GLP-1 (7-39); Arg∞^Lys^-GLP-l (7-40); Arg 26 Lys 36 - GLP-1(7-37); Arg 34 Lys 36 -GLP-1(7-37); Arg 26 Lys 39 -GLP-1 (7-39); Arg 34 Lys 40 -GLP-1(7-40); Arg 2634 Lys 36 ' 39 -GLP-1 (7-39); Arg∞^Lys∞^-GLP-l (7-40); Gly 8 Arg26-GLP-1 (7-37); Gly^rg^-GLP-I (7-37); Glylys∞-GLP-I (7-37); Gly'Αrg^Lys -GLP-l (7-37); 56 G i y 8 Arg 26,3 4Lys 39 . GL p. 1 (7 .3 9) . Gly 8 Arg 263 Lys 40 -GLP-1 (7-40); Gly 8 Arg 26 Lys 36 -GLP-1 (7-37); GI Arg^Lys∞-GLP-l (7-37); Gly 8 Arg 26 Lys 39 -GLP-1 (7-39); GI Arg^Lys^-GLP-l (7-40); Gly 8 Arg 263 Lys 3639 -GLP-1 (7-39) and Gly 8 Arg 2634 Lys 36 ' 40 -GLP-1 (7-40). 34. A pharmaceutical composition according to any of the claims 1 to 32, comprising a GLP-1 derivative, wherein the parent peptide is selected from the group comprising Arg 634 Lys 38 GLP-1 7-38) Arg 2634 Lys 39 GLP-1(7-39) Arg 26 ' 34 Lys 40 GLP-1(7-40); Arg 2634 Lys 41 GLP-1 7-41) Arg 2634 Lys 42 GLP-1 (7-42) Arg∞^Lys^GLP-l 7-43) Arg^Lys^GLP-l 7-44) Arg 26 ' 34 Lys 45 GLP-1(7-45) Arg 2634 Lys 38 GLP-1 1-36) Arg 2634 Lys 39 GLP-1 1-39) Arg^Lys^GLP-IO^O) Arg 2634 Lys 1 GLP-1 1-41) Arg 2634 Lys 42 GLP-1 1-42) Arg 2634 Lys 43 GLP-1(1-43) Arg ^Lys^GLP-l 1-44) Arg 2634 Lys 45 GLP-1 1-45) Arg 26 ' 34 Lys 38 GLP-1(2-38) Arg 2634 Lys 39 GLP-1 2-39) Arg 263 Lys 40 GLP-1 2-40) Arg 2634 Lys 41 GLP-1(2-41) Arg 2634 Lys 42 GLP-1 2-42) Arg 2634 Lys 43 GLP-1 2-43) Arg 263 Lys 4 GLP-1(2-44) Arg 2634 Lys 45 GLP-1 2-45) Arg 2W4 Lys 3β GLP-1 3-38) Arg 2634 Lys 39 GLP-1(3-39) Arg∞^Lys^GLP-l 3-40) Arg 2634 Lys 41 GLP-1 3-41) Arg 2634 Lys 2 GLP-1(3-42) Arg 2634 Lys 43 GLP-1 3-43) Arg∞^Lys^GLP-l 3-44) Arg ^Lys^GLP-l β^δ) Arg 2634 Lys 38 GLP-1 4-38) Arg 2634 Lys 39 GLP-1 4-39) Arg 26 ' 34 Lys 40 GLP-1(4-40) Arg 2634 Lys 41 GLP-1 4-41) Arg M4 Lys 42 GLP-1 4-42) Arg 2634 Lys 43 GLP-1(4-43) Arg^Lys^GLP-l 4-44) Arg 2634 Lys 45 GLP-1 4-45) Arg^^Lys∞GLP-^δ-Sδ) Arg 2634 Lys 39 GLP-1 5-39) Arg^Lys^GLP-l 5-40) Arg∞^Lys^GLP-l^l) Arg 2634 Lys 42 GLP-1 5-42) Arg 2634 Lys 43 GLP-1 5-43) Arg 2634 Lys 44 GLP-1(5-44) Arg∞^Lys^GLP-l 5-45) Arg 2M4 Lys aβ GLP-1 6-38) Arg 2634 Lys 39 GLP-1(6-39) Arg 2634 Lys 40 GLP-1 6-40) Arg 263 Lys 41 GLP-1 6-41) Arg 2634 Lys 4 GLP-1(6-42) Arg 2634 Lys 43 GLP-1 6-43) Arg∞^Lys^GLP-l 6-44) Arg∞^Lys^GLP-I^S); Arg 26 Lys 38 GLp . 1 (1 _ 38) . Arg^Lys^GLP- 1(1-38); Arg ^Lys∞∞GLP-IO-Sδ); Arg∞Lys∞GLP-l (7-38); Arg^Lys∞GLP-l (7-36); Arg∞^Lys∞∞GLP-l (7-38); Arg∞^Lys∞GLP-l (7-38); Arg 26 Lys 39 GLP-1 (1-39); Arg^Lys∞GLP-l (1-39); Arg∞^Lys ∞GLP-l (1-39); Arg 26 Lys 39 GLP-1 (7-39); Arg 34 Lys 39 GLP-1(7-39) and Arg∞^Lys∞ GLP-l (7-39). 35. A method for improving the solubility and/or stability of GLP-1 or a fragment or an analogue thereof, characterised in that a lipophilic substituent is introduced on any one of the amino acid residues of the parent peptide. 36. A method according to claim 35, wherein a lipophilic substituent is introduced on any one of the amino acid residues in position 1δ-3δ, preferably 26-34. 57 37. A method according to claim 35 or 36, wherein the lipophilic substituent comprises from 4 to 40 carbon atoms, preferably from 3 to 25 carbon atoms. 36. A method according to any one of claim 35 to 37, wherein the lipophilic substituent is the acyl group of a straight-chain or branched fatty acid. 39. A method according to any one of claim 36, wherein the acyl group is selected from the group comprising CH 3 (CH 2 ) n CO-, wherein n is 4 to 38, preferably CH 3 (CH 2 ) 6 CO-, CH 3 (CH 2 ) 8 CO-, CH 3 (CH 2 ) 10 CO-, CH 3 (CH 2 ) 12 CO-, CH 3 (CH 2 ) 14 CO-, CH 3 (CH 2 ) 16 CO-, CH 3 (CH 2 ) 18 CO-, CH 3 (CH 2 ) 20 CO- and CH 3 (CH 2 ) 22 CO-. 40. A method according to any one of claims 35 to 39, wherein the GLP-1 is GLP-1 (A-B) whe- rein A is an integer from 1 to 7 and B is an integer from 38 to 45, or an analogue thereof. 41. A method according to claim 40, wherein the GLP-1 is 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); and analogues thereof. 42. A method according to claim 40, wherein the GLP-1 is selected from the group comprising GLP-1(1-35); GLP-1(1-36); GLP-1(1-36)amide; GLP-1(1-37); GLP-1(1-33); GLP-1(1-39); GLP-1(1-40); GLP-1(1-41); and an analogues thereof.

Inventors: BJOERN SOEREN ERIK, HUUSFELDT PER OLAF, KAARSHOLM NIELS C, KNUDSEN LISELOTTE BJERRE, NIELSEN PER FRANKLIN, OLSEN HELLE BIRK, BJOERN, SOEREN, ERIK, HUUSFELDT, PER, OLAF, KAARSHOLM, NIELS, C., KNUDSEN, LISELOTTE, BJERRE, NIELSEN, PER, FRANKLIN, OLSEN, HELLE, BIRK
Applicants: NOVONORDISK AS, NOVO NORDISK A/S
Application year: 1999
Main classification: A61K38
IPC codes: A61K38, A61K47, A61P3, A61P43, C07K14