a2, United States Patent
`US 6,458,924 B2
`(10) Patent No.:
`*Oct. 1, 2002
`(45) Date of Patent:
`Knudsenetal.
`
`US006458924B2
`
`(54) DERIVATIVES OF GLP-1 ANALOGS
`
`(75)
`
`5,614,492 A
`5,869,602 A *
`5,912,229 A
`
`3/1997 Habener ....... cee 514/12
`
`.... 530/308
`2/1999 Jonassen....
`6/1999 Thim et al. 0. 514/12
`
`FOREIGN PATENT DOCUMENTS
`
`Inventors: Liselotte Bjerre Knudsen, Valby (DK);
`Per Olaf Huusfeldt, Kobenhavn K
`(DK); Per Franklin Nielsen, Verlgse
`EP 0 619 322 A2=10/1994
`
`(DK)
`EP
`0 658 568
`6/1995
`EP
`0 708 179
`4/1996
`GB
`1 202 607
`8/1970
`WO
`WO 87/06941
`11/1987
`WO
`WO 90/11296
`10/1990
`WO
`WO 91/11457
`8/1991
`WO
`WO 95/07931
`3/1995
`WO
`WO 95/31214
`11/1995
`WO
`WO 95/32730
`12/1995
`WO
`96/29342
`9/1996
`WO
`WO 96/29342
`9/1996
`WO
`WO 96/29344
`9/1996
`WO
`WO 97/31943
`9/1997
`WO
`WO 98/08871
`3/1998
`WO
`98/08871
`3/1998
`WO
`WO 98/08873
`3/1998
`WO
`WO 98/08531
`5/1998
`WO
`WO 98/19698
`5/1998
`
`(73) Assignee: Novo Nordisk A/S, Bagsvaerd (DK)
`
`(*) Notice:
`
`Subject to any disclaimer, the term ofthis
`patent is extended or adjusted under 35
`US.C. 154(b) by 0 days.
`
`This patent is subject to a terminal dis-
`claimer.
`
`(21) Appl. No.: 09/398,111
`
`(22)
`
`Filed:
`
`Sep. 16, 1999
`
`Related U.S. Application Data
`
`(60)
`
`(63) Continuation-in-part of application No. 09/265,141, filed on
`Mar. 8, 1999, now Pat. No. 6,384,016, and a continuation-
`in-part of application No. 09/258,750, filed on Feb. 26,
`1999, now Pat. No. 6,268,343, which is a continuation-in-
`part of application No. 09/038,432, filed on Mar. 11, 1998,
`now abandoned, whichis a continuation-in-part of applica-
`tion No. 08/918,810, filed as application No. PCT/DK97/
`00340 on Aug. 22, 1997, now abandoned.
`Provisional application No. 60/035,904, filed on Jan. 24,
`1997, provisional application No. 60/036,226,filed on Jan.
`25, 1997, provisional application No. 60/036,255, filed on
`Jan. 24, 1997, provisional application No. 60/078,422,filed
`on Mar. 18, 1998, provisional application No. 60/082,478,
`filed on Apr. 21, 1998, provisional application No. 60/082,
`479, filed on Apr. 21, 1998, provisional application No.
`60/082,480, filed on Apr. 21, 1998, provisional application
`No. 60/082,802, filed on Apr. 23, 1998, and provisional
`application No. 60/084,357, filed on May 5, 1998.
`
`(30)
`
`Foreign Application Priority Data
`
`.. 0263/98 Feb. 27, 1998
`
`Aug. 30, 1996
`Nov. 8, 1996
`Dec. 20, 1996
`Feb. 27, 1998
`
`Feb. 27, 1998
`Feb. 27, 1998
`Feb. 27, 1998
`Mar. 13, 1998
`Apr. 8, 1998
`Apr. 8, 1998
`Apr. 8, 1998
`
`(DK) cececcsscesssssessesstessesseessssseessessees 0931/96
`(DK) cesecsssesssssessssssesssesseesesesesssessees 1259/96
`(DK) cecessessesssessesssessssseesesssessusssseses 1470/96
`(DK) ....
`... 0264/98
`(DK)....
`... 0268/98
`(DK)....
`(DK) vecescessesstesssesssessnsessessnessteenies 0272/98
`(DK) vecescesssesstessseessessnsessessnessteenies 0274/98
`(EP) .....
`..- 98610006
`(DK) cecsccsscssesssessssssesssesseestsssessnessees 0508/98
`(DK) cecsccsscssesssessessesssesseesesesesssessess 0509/98
`(DK)....
`... 1998 00507
`
`
`
`Int. Ch? eee A61K 38/16; A61K 38/26
`(51)
`(52) US. Ch. ect eeneeeees 530/324; 530/345; 514/2;
`514/12
`(58) Field of Search... 514/2, 12; 530/324,
`530/345
`
`(56)
`
`References Cited
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`
`*
`
`OTHER PUBLICATIONS
`
`Broderick, Diabetologia (1995) vol. 38, No. Suppl. 1, pp.
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`Association for the Study of Diabetes Stockholm, Sweden
`Sep. 12-16, 1995.*
`M. Gutniaket al., Antidiabetogenic Effect of Glucagon—Like
`Peptide (7-36) Amide in Normal Subjects and Patients with
`Diabetes with Diabetes Mellitus.
`
`M. Navarro et al., Changes in Food Intake Induced by
`GLP-1(7-36) Amide In the Rat, Abstracts of the 15”
`International Diabetes Federation Congress, Nov. 6-11,
`1194 Kobe, poster presentation 11A 5PP1295 Issued 1994.
`R. Schick et al., “Glucagon—like peptide 1-a novel brain
`peptide Involved in feeding regulation “Obesity in Europe
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`P.D. Lambert et al., “A Role for GLP-1(7-36)NH, in the
`Central Control Of Feeding Behavior” Digestion 1994; vol.
`54. pp. 360-361.
`B. Willms et al., “Gastric Emptying, Glucose Responses,
`and Insulin Secretion after a Liquid Test Meal:Effects of
`Exogenous Glucagon—-Like Peptide-1 (GLP-1)
`(7-36)
`Amide
`in Type
`2
`(Noninsulin—-Dependent) Diabetic
`Patients” Journal of Clinical Endocrinology and Metabolism
`vol. 8 No. 1 (1996) pp. 327-332.
`
`(List continued on next page.)
`
`Primary Examiner—Christopher S. F. Low
`Assistant Examiner—David Lukton
`(74) Attorney, Agent, or Firm—Reza Green, Esq.; Richard
`Bork, Esq.
`
`(57)
`
`ABSTRACT
`
`The present invention relates to a pharmaceutical composi-
`tion comprising a GLP-1 derivative having a lipophilic
`substituent; and a surfactant.
`
`20 Claims, 1 Drawing Sheet
`
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`

`US 6,458,924 B2
`Page 2
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`Woodset al., “Signals that regulate food intake and energy
`Homeostasis”. Science, 280:1378-1383, May 29, 1998.
`Thorens T. “Glucagon-like peptide—1 and control of insulin
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`Henriksen et al. Peptide amidation by chemical protein
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`Wang et al. “Glucagon—like peptide—-1 is a physiological
`incretin in rat”. J. Clin. Invest., Jan 1995) (1) 417-21.
`Bell et al. “Exon duplication and divergence in the human
`Preproglucagon gene”. Nature, (Jul. 28—Aug. 3, 1983).
`Wettergren
`et
`al.
`“Truncated GLP-1
`(proglucagon
`78-107—amide) inhibits gastric and pancreatic functions in
`man”. Digest. Dis. Sci., (Apr. 1993) 38 (4) 665-73.
`Suzuki et al. “Comparison of the effects of various C-ter-
`minal and N-terminal fragment peptides of glucagons—like
`peptide—1 on insulin and glucagonsrelease from the isolat-
`edcx perfused rat pancreas”. Endocrinology, (Dec. 1989)
`125(6) 3109-14.
`
`Navarroet al., Journal of Neurochemistry, vol. 67, No. 5, pp.
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`Clodfelter et al., (1998) Pharmaceutical Res. 15(2):254—262.
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`244, No. 24, Dec. 25, 1969, pp. 6675-6679.
`Sasaki et al., “X-Ray Analysis of Glucagon andIts Rela-
`tionship to Receptor Binding”, Nature Vo. 257, Oct. 30,
`1975, pp. 751-757.
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`land Biomedical Press, vol. 119, No. 2, Oct. 1980, pp.
`265-270.
`
`Epand et al., “Molecular Interactions In The Model Lipo-
`protein Complex Formed Between Glucagon and Dimyris-
`toylglycerophosphocholine”, Biochemistry vol. 16, No. 20,
`1977.
`
`Schneider et al., “Polypeptide HormoneInteraction” (Glu-
`cagon Binding To Lysolecithin), The Journal of Biological
`Chemistry, Vo. 247, No. 16, Aug. 25, 1972, pp. 4986-4991.
`Schneideret al., “Polypeptide Hormone Interaction” (Con-
`formational Changes of Glucagon Bound To Lysolecithin),
`The Journal of Biological Chemistry, Vo. 247, No. 16, Aug.
`25, 1972, pp. 4992-4995.
`Robinsonetal., “Lipid—Induced Conformational Changesin
`Glucagon, Secretin, and Vasoactive Intestinal Peptide”,
`Biopolymers, vol. 21, 1982, pp. 1217-1228.
`Hamedet al., “Bahavior ofAmphipathic Helices on Analysis
`Via Matrix Methods, With Application to Glucagon, Secre-
`tin, and Vasoactive Intestinal Peptide”, Biopolymers, vol.
`22, 1983, pp. 1003-1021.
`Wuet al., “Helical Conformation of Glucagon in Surfactant
`Solutions”, Americal Chemical
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`2117-2122.
`
`Bosch et al., “Physicochemical Characterization of Glu-
`cagon—Containing Lipid Micelles” Biochimic et Biophysica
`Acta, 603 (1980) pp. 298-312.
`Thornton et al., Structure of Glucagon—Like Peptide(7-36)
`Amide in a Dodecylphosphocholine Micelle as Determined
`by 2D NMR,Biochemistry 1994, 33, pp. 3532-3539.
`
`* cited by examiner
`
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`

`U.S. Patent
`
`Oct. 1, 2002
`
`US 6,458,924 B2
`
`
`
`0 f
`
`-4
`
`
`
`
`
`
`
`
`
`
`—+-—g
`
`-o- b
`
`--A--
`
`Cc
`
`a |
`
`—V— gip-1(7-37)
`
`—O-— e
`
`—-O--
`
`f
`
`cp el cl el onl oy,
`
`--W-- g
`
`0.1
`
`1
`
`10
`
`100
`
`1000
`
`[peptide] (uM)
`
`FIG. 1
`
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`US 6,458,924 B2
`
`1
`DERIVATIVES OF GLP-1 ANALOGS
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`
`This application is a continuation-in-part of Ser. No.
`09/265,141 filed Mar. 8, 1999 now US. Pat. No. 6,384,016
`and of Ser. No. 09/258,750 filed Feb. 26, 1999 now USS. Pat.
`No. 6,268,343 which is a continuation-in-part of Ser. No.
`09/038,432 filed Mar. 11, 1998 now abandoned whichis a
`continuation-in-part of Ser. No. 08/918,810 filed Aug. 26,
`1997 now abandoned, which is a 371 and of PCTapplication
`Ser. No. PCT/DK97/00340 filed Aug. 22, 1997, and claims
`priority of U.S. provisional application Ser. Nos. 60/035,
`904, 60/036,226, 60/036,255, 60/078,422, 60/082,478,
`60/082,479, 60/082,480, 60/082,802, and 60/084,357 filed
`Jan. 24, 1997, Jan. 25, 1997, Jan. 24, 1997, Mar. 18, 1998,
`Apr. 21, 1998, Apr. 21, 1998, Apr. 21, 1998, Apr. 23, 1998,
`and May 5, 1998, respectively, and of Danish application
`serial nos. 0931/96, 1259/96, 1470/96, 0263/98, 0264/98,
`0268/98, 0272/98, 0274/98, 0507/98, 0508/98, and 0509/98
`filed Aug. 30, 1996, Nov. 8, 1996, Dec. 20, 1996, Feb. 27,
`1998, Feb. 27, 1998, Feb. 27, 1998, Feb. 27, 1998, Feb. 27,
`1998, Apr. 8, 1998, Apr. 8, 1998 and Apr. 8, 1998,
`respectively, and of European application no. 98610006.3
`filed Mar. 13, 1998, the contents of each of which is fully
`incorporated herein by reference.
`FIELD OF THE INVENTION
`
`invention relates to novel derivatives of
`The present
`human glucagon-like peptide-1 (GLP-1) and fragments and/
`or analogues thereof which have a protracted profile of
`action and to methods of making and using them.
`BACKGROUND OF THE INVENTION
`
`Peptides are widely used in medical practice, and since
`they can be produced by recombinant DNA technologyit
`can be expected that their importance will increase also in
`the years to come. When native peptides or analogues
`thereof are used in therapy it is generally found that they
`have a high clearance. A high clearance of a therapeutic
`agent is inconvenient in cases whereit is desired to maintain
`a high blood level thereof over a prolonged period of time
`since repeated administrations will
`then be necessary.
`Examples of peptides which have a high clearance are:
`ACTH, corticotropin-releasing factor, angiotensin,
`calcitonin,
`insulin, glucagon, glucagon-like peptide-1,
`glucagon-like peptide-2,
`insulin-like growth factor-1,
`insulin-like growth factor-2, gastric inhibitory peptide,
`growth hormone-releasing factor, pituitary adenylate
`cyclase activating peptide, secretin, enterogastrin,
`somatostatin, somatotropin, somatomedin, parathyroid
`hormone,
`thrombopoietin, erythropoietin, hypothalamic
`releasing factors, prolactin, thyroid stimulating hormones,
`endorphins, enkephalins, vasopressin, oxytocin, opiods and
`analogues thereof, superoxide dismutase,
`interferon,
`asparaginase, arginase, arginine deaminase, adenosine
`deaminase and ribonuclease. In some casesit is possible to
`influence the release profile of peptides by applying suitable
`pharmaceutical compositions, but this approach has various
`shortcomings and is not generally applicable.
`The hormonesregulating insulin secretion belong to the
`so-called enteroinsular axis, designating 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
`
`2
`so-called incretin effect, is probably essential for a normal
`glucose tolerance. Many of the gastrointestinal hormones,
`including gastrin and secretin (cholecystokininis not insuli-
`notropic in man), are insulinotropic, but the only physiologi-
`cally important ones,
`those that are responsible for the
`incretin effect, are the glucose-dependent
`insulinotropic
`polypeptide, GIP, and glucagon-like peptide-1(GLP-1).
`Becauseof its insulinotropic effect, GIP, isolated in 1973 (1)
`immediately attracted considerable interest among diabe-
`tologists. However, numerous investigations carried out
`during the following years clearly indicated that a defective
`secretion of GIP was not involved 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 insulinotropic 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 gastrointes-
`tinal secretion and metabolism (5). The glucagon geneis
`processed differently in the pancreas and in the intestine. In
`the pancreas (9), the processing leads to the formation and
`parallel secretion of 1) glucagonitself, occupying positions
`33-61 of proglucagon (PG); 2) an N-terminal peptide of 30
`amino acids (PG (1-30)) often called glicentin-related pan-
`creatic peptide, GRPP (10, 11); 3) a hexapeptide correspond-
`ing 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,
`whichis 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 conspicuousbiological activi-
`ties.
`
`Being secreted in parallel with glicentin/enteroglucagon,
`it follows that the manystudies of enteroglucagon secretion
`(6, 7) to some extent also apply to GLP-1 secretion, but
`GLP-1 is metabolised more quickly with a plasmahalf-life
`in humans of 2 min (19). Carbohydrate or fat-rich meals
`stimulate secretion (20), presumably as a result of direct
`interaction of yet unabsorbednutrients with the microvilli of
`the open-type L-cells of the gut mucosa. Endocrine or neural
`mechanisms promoting GLP-1 secretion may exist but have
`not yet been demonstrated in humans.
`The incretin function of GLP-1(29-31) has been clearly
`illustrated in experiments with the GLP-1 receptor
`antagonist, exendin 9-39, which dramatically reduces the
`
`wn
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`10
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`15
`
`25
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`30
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`35
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`40
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`60
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`65
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`US 6,458,924 B2
`
`3
`incretin effect elicited by oral glucose in rats (21, 22). The
`hormone interacts directly with the Bcells via the GLP-1
`receptor (23) which belongs to the glucagon/VIP/calcitonin
`family of G-protein-coupled 7-transmembrane spanning
`receptors. The importance of the GLP-1 receptor in regu-
`lating insulin secretion wasillustrated in recent experiments
`in which a targeted disruption of the GLP-1 receptor gene
`was carried out
`in mice. Animals homozygous for the
`disruption had greatly deteriorated glucose tolerance and
`fasting hyperglycaemia, and even heterozygous animals
`were glucoseintolerant (24). The signal transduction mecha-
`nism (25) primarily involves activation of adenylate cyclase,
`but elevations of intracellular Ca?* are also essential (25,
`26). The action of the hormone is best described as a
`potentiation of glucose stimulated insulin release (25), but
`the mechanism that couples glucose and GLP-1 stimulation
`is not known. It may involve a calcium-induced calcium
`release (26, 27). As already mentioned, the insulinotropic
`action of GLP-1 is preserved in diabetic B-cells. The relation
`of the latter to its ability to convey “glucose competence”to
`isolated insulin-secreting cells (26, 28), which respond
`poorly to glucose or GLP-1 alone, but fully to a combination
`of the two, is also not known. Equally importantly, however,
`the hormonealso potently inhibits glucagon secretion (29).
`The mechanism is not known,but seemsto be paracrine, via
`neighbouring insulin or somatostatin cells (25). Also the
`glucagonostatic action is glucose-dependent, so that
`the
`inhibitory effect decreases as blood glucose decreases.
`Because of this dual effect, if the plasma GLP-1 concentra-
`tions increase either by increased secretion or by exogenous
`infusion the molar ratio of insulin to glucagon in the blood
`that reaches the liver via the portal circulation is greatly
`increased, whereby hepatic glucose production decreases
`(30). As a result blood glucose concentrations decrease.
`Because of the glucose dependencyofthe insulinotropic and
`glucagonostatic actions, the glucose lowering effect is self-
`limiting, and the hormone,
`therefore, does not cause
`hypoglycaemia regardless of dose (31). The effects are
`preserved in patients with diabetes mellitus (32), in whom
`infusionsof slightly supraphysiological doses of GLP-1 may
`completely normalise blood glucose values in spite of poor
`metabolic control and secondary failure to sulphonylurea
`(33). The importance of the glucagonostatic effect is illus-
`trated by the finding that GLP-1 also lowers blood glucose
`in type-i diabetic patients without residual B-cell secretory
`capacity (34).
`In addition to its effects on the pancreatic islets, GLP-1
`has powerful actions on the gastrointestinal tract. Infused in
`physiological amounts, GLP-1 potently inhibits
`pentagastrin-induced as well as meal-induced gastric acid
`secretion (35, 36). It also inhibits gastric emptying rate and
`pancreatic enzymesecretion (36). Similar inhibitory effects
`on gastric and pancreatic secretion and motility may be
`elicited in humans upon perfusion of the ileum with
`carbohydrate- or
`lipid-containing solutions (37, 38).
`Concomitantly, GLP-1 secretion is greatly stimulated, and it
`has been speculated that GLP-1 may be at
`least partly
`responsible for this so-called “ileal-brake” effect (38). In
`fact, recent studies suggest that, physiologically, the ileal-
`brake effects of GLP-1 may be more important than its
`effects on the pancreatic islets. Thus,
`in dose response
`studies GLP-1 influences gastric emptying rate at infusion
`rates at least as low as those required to influence islet
`secretion (39).
`GLP-1 seemsto have an effect on food intake. Intraven-
`tricular administration of GLP-1 profoundly inhibits food
`intake in rats (40, 42). This effect seems to be highly
`
`4
`specific. Thus, N-terminally extended GLP-1(PG 72-107)
`amide is inactive and appropriate doses of the GLP-1
`antagonist, exendin 9-39, abolish the effects of GLP-1(41).
`Acute, peripheral administration of GLP-1 does not inhibit
`food intake acutely in rats (41, 42). However, it remains
`possible that GLP-1 secreted from the intestinal L-cells may
`also act as a satiety signal.
`Not only the insulinotropic effects but also the effects of
`GLP-1 on the gastrointestinal tract are preserved in diabetic
`patients (43), and may help curtailing meal-induced glucose
`excursions, but, more importantly, may also influence food
`intake. Administered intravenously, continuously for one
`week, GLP-1 at 4 ng/kg/min has been demonstrated to
`dramatically improve glycaemic control in NIDDM patients
`without significant side effects (44). The peptide is fully
`active after subcutaneous administration (45), but is rapidly
`degraded mainly due to degradation by dipeptidyl peptidase
`IV-like enzymes (46, 47).
`The amino acid sequence of GLP-1 is given i.a. by
`Schmidt et al. (Diabetologia 28 704-707 (1985). Human
`GLP-1 is a 37 amino acid residue peptide originating from
`preproglucagon which is synthesised, 1.a. in the L-cells in
`the distal ileum, in the pancreas and in the brain. Processing
`of preproglucagon to GLP-1(7—-36)amide, GLP-1(7-37) and
`GLP-2 occurs mainly in the L-cells. Although theinteresting
`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 com-
`pounds whose plasmaprofile were highly protracted andstill
`had retained activity.
`GLP-1 and analogues of GLP-1 and fragments thereof are
`useful 1.a. in the treatment of Type 1 and Type 2 diabetes and
`obesity.
`including
`WO 87/06941 discloses GLP-1 fragments,
`GLP-1(7-37), and functional derivatives thereof and to their
`use as an insulinotropic agent.
`including
`WO 90/11296 discloses GLP-1 fragments,
`GLP-1(7-36), and functional derivatives thereof which have
`an insulinotropic activity which exceeds the insulinotropic
`activity of GLP-1(1-36) or GLP-1(1-37) andto their use as
`insulinotropic agents.
`The amino acid sequence of GLP-1(7-36) and GLP-1
`(7-37) is (SEQ ID NO: 1):
`
`17
`16
`#15
`14
`13
`12
`11
`10
`9
`8
`7
`His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-
`18
`#19
`20
`21
`22
`23
`24
`25
`26
`27
`28
`Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-
`29
`30
`31
`32
`33
`34
`#35
`36
`Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-xX
`
`(T)
`
`wherein X is NH, for GLP-1(7-36) and X is Gly for
`GLP-1(7-37).
`WO 91/11457 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.
`EP 0708179-A2 (Eli Lilly & Co.) discloses GLP-1 ana-
`logues and derivatives that include an N-terminal imidazole
`group and optionally an unbranched C,-C,, acyl group in
`attached to the lysine residue in position 34.
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`35
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`40
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`45
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`50
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`55
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`60
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`65
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`US 6,458,924 B2
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`5
`EP 0699686-A2 (Eli Lilly & Co.) discloses certain
`N-terminal truncated fragments of GLP-1 that are reported
`to be biologically active.
`Unfortunately, the high clearance limits the usefulness of
`these compounds. Thus there still is a need for improve-
`ments in this field.
`Accordingly, it is an object of the present invention to
`provide derivatives of GLP-1 and analogues thereof which
`have a protracted profile of action relative to GLP-1(7-37).
`It is a further object of the invention to provide derivatives
`of GLP-1 and analogues thereof which have a lowerclear-
`ance than GLP-1(7-37).
`It is a further object of the invention to provide a phar-
`maceutical composition with improved solubility and sta-
`bility.
`
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