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WO2008132477A1 - Composés et leurs utilisations - Google Patents

Composés et leurs utilisations Download PDF

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Publication number
WO2008132477A1
WO2008132477A1 PCT/GB2008/001489 GB2008001489W WO2008132477A1 WO 2008132477 A1 WO2008132477 A1 WO 2008132477A1 GB 2008001489 W GB2008001489 W GB 2008001489W WO 2008132477 A1 WO2008132477 A1 WO 2008132477A1
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Prior art keywords
compound according
group
ala
alkyl
glp
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PCT/GB2008/001489
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English (en)
Inventor
Patrick Kanda
Peter Laing
Ram Prakash Sharma
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Activotec SPP Ltd
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Activotec SPP Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/26—Glucagons
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/605—Glucagons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/02—Non-specific cardiovascular stimulants, e.g. drugs for syncope, antihypotensives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/13—Labelling of peptides

Definitions

  • the present invention relates to certain compounds, to processes for their preparation, compositions comprising them and methods of treatment of the human or animal body employing them.
  • Type Il diabetes non-insulin dependent diabetes mellitus - NIDDM
  • a non-insulin dependent diabetes mellitus - NIDDM is a condition characterized by a resistance to insulin action in peripheral tissues such as muscle, adipose and liver, and by a progressive failure of the ability of the islet ⁇ -cells to secrete insulin.
  • current therapies do not halt the progression of ⁇ -cell failure, virtually all NIDDM patients eventually require insulin to control blood glucose levels.
  • the most commonly prescribed therapeutics for such patients are the sulfonylureas, a class of drugs that stimulate insulin secretion.
  • 10-20% of the patients receiving sulfonylurea therapy fail to maintain acceptable blood glucose levels, and switch to insulin therapy.
  • Insulin therapy is undesirable from a variety of points of view. Firstly, it has a narrow therapeutic index. This leads to poor control of blood glucose levels, since most patients and physicians err in favour of high glucose levels rather than risk hypoglycaemia and coma.
  • the factors to be taken into account include the amount of food consumed, the interval between meals, the amount of physical exercise, and the prevailing blood glucose level (the determination of which requires blood glucose monitoring).
  • the hormone glucagon-like peptide-1 (7-37) (GLP-1) is released by intestinal L cells in response to ingested nutrients and acts to promote glucose-dependent insulin secretion ensuring efficient postprandial glucose homeostasis.
  • This hormone binds the GLP-1 receptor present on a number target tissues including the lungs, pancreatic ⁇ cells, brain, muscle, and gut, achieving normal glucose levels in five separate but complementary ways.
  • the peptide is synthesised as a prohormone in which the N terminal 6 amino acids are processed to yield the biologically active GLP-1 7-37 sequence:
  • DPP IV cleavage occurs between AIa 8 and GIu 9 , completely inactivating it and resulting in a circulating half-life of less than 2 minutes for the active form of GLP-1.
  • Various strategies have been employed to protect GLP-1 against both renal clearance and DPP-IV cleavage in efforts to produce a more stable, enduring anti- diabetic drug. They include amino acid substitutions at AIa 8 and elsewhere, covalent attachment of fatty acids, and the linking of GLP-1 to albumin. Modification of GLP-1 through fatty acid attachment confers protection from renal clearance and DPP-IV activity from its resulting association in the bloodstream with albumin. A therapeutic candidate for treating Type Il diabetes based on this modification has emerged and is currently undergoing clinical trials. In addition, a number of small molecule DPP IV inhibitors are being developed for oral delivery to prolong the activity of endogenously secreted GLP-1.
  • Ri is selected from the group consisting of His, D-histidine, desamino-histidine, 2-amino-histidine, ⁇ -hydroxy- histidine, homohistidine, ⁇ -fluoromethyl-histidine, and ⁇ -methyl-histidine
  • X is selected from the group consisting of Met, Asp, Lys, Thr, Leu, Asn, GIn, Phe, VaI, and Tyr
  • Y and Z are independently selected from the group consisting of GIu, GIn 1 Ala, Thr, Ser, and GIy
  • R 2 is selected from the group consisting of NH 2 , and GIy- OH; provided that, if R 1 is His, X is VaI, Y is GIu, and Z is GIu, then R 2 is NH 2 .
  • the GLP-1 analogues are stated to have an increased duration of action and resistance to DPP-IV.
  • Z 1 is substituent of the terminal amino group of the peptide
  • Z 2 is a substituent of the terminal carbonyl group of the peptide
  • X 1 to X 14 each represents, independently of the others a natural or non-natural amino acid residue, having the D or L configuration.
  • the compounds are stated to have agonist character in relation to f GLP-1 receptors, in addition to increased metabolic stability and duration of action when compared to the natural peptide.
  • Ri is selected from the group consisting of L-histidine, D-histidine, desamino-histidine, 2-amino-histidine, ⁇ -hydroxy-histidine, homohistidine, ⁇ - fluoromethyl-histidine, and ⁇ -methyl-histidine;
  • X is selected from the group consisting of Ala, GIy, VaI, Thr, lie, and alpha-methyl-Ala;
  • Y is selected from the group consisting of GIu, GIn, Ala, Thr, Ser, and GIy;
  • Z is selected from the group consisting of GIu, GIn, Ala, Thr, Ser, and GIy;
  • R 2 is selected from the group consisting of NH 2 , and GIy-OH; providing that the compound has an isoelectric point in the range from about 6.0 to about 9.0 and further providing that when R 1 is His, X is Ala, Y is GIu, and Z is GIu, R 2 must be
  • the peptides are said to have increased stability to storage and in vivo.
  • US6849708 discloses insulinotropic peptides comprising a fragment of GLP-1 and derivatives thereof. A problem that remains is the provision of therapeutic agents for the treatment of diabetes.
  • a further problem that remains is the provision of compounds having GLP-1 like activity while having superior stability compared with naturally occurring GLP-1.
  • the present invention addresses problems of the prior art.
  • a compound of the invention covalently linked to a carrier molecule ("complexed compound").
  • a pharmaceutical composition comprising a compound or complexed compound of the invention above together with a pharmaceutically acceptable carrier or excipient.
  • a method of treatment of a human or animal suffering from a condition selected from hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, atheroschlerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcers comprising administering to said mammal a compound or complexed compound of the invention or a composition comprising such a compound.
  • a compound or of a complexed compound of the invention or a composition comprising such a compound for the treatment of a condition selected from hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, 8 001489
  • the compounds of the invention comprise a moiety having GLP-1 like activity.
  • the term "having GLP-1 like activity” refers to a moiety which in isolation (i.e. not attached to the linker or aminothiol moiety) has activity of at least 10 % of that exhibited by GLP-1 (7-37) in a binding assay.
  • the moiety has activity of at least 10 % of that exhibited by GLP-1 (7-37) in a binding assay. More preferably, the moiety has activity of at least 20 % of that exhibited by GLP-1 (7-37) in a binding assay. More preferably, the moiety has activity of at least 50 % of that exhibited by GLP-1 (7-37) in a binding assay. More preferably, the moiety has activity of at least 90 % of that exhibited by GLP-1 (7-37) in a binding assay. More preferably, the moiety has activity of at least equal to that exhibited by GLP-1 (7-37) in a binding assay.
  • Examples of specific moieties having GLP-1 like activity are polypeptides comprising the 1-37 amino acid sequence of GLP-1 , viz. formula I:
  • polypeptides comprising the 7-37 amino acid sequence of GLP-1, viz. formula II:
  • polypeptides comprising the 7-36 amino acid sequence of GLP-1 , viz. formula 111:
  • Ri is selected from the group consisting of His, D-histidine, desamino-histidine, 2-amino-histidine, ⁇ -hydroxy- histidine, homohistidine, ⁇ -fluoromethyl-histidine, and ⁇ -methyl-histidine;
  • X is selected from the group consisting of Met, Asp, Lys, Thr, Leu, Asn, GIn, Phe, VaI, and Tyr;
  • Y and Z are independently selected from the group consisting of GIu, GIn, Ala, Thr, Ser, and GIy, and R 2 is selected from the group consisting of NH 2 , and GIy-;
  • Zi is substituent of the terminal amino group of the peptide
  • Z 2 is a substituent of the terminal carbonyl group of the peptide
  • X 1 to X 14 each represents, independently of the others a natural or non-natural amino acid residue, having the D or L configuration.
  • R 1 is selected from the group consisting of L-histidine, D-histidine, desamino-histidine, 2-amino-histidine, ⁇ -hydroxy-histidine, homohistidine, ⁇ - fluoromethyl-histidine, and ⁇ -methyl-histidine;
  • X is selected from the group consisting of Ala, GIy, VaI, Thr, lie, and alpha-methyl-Ala;
  • Y is selected from the group consisting of GIu 1 GIn, Ala, Thr, Ser, and GIy;
  • Z is selected from the group consisting of GIu, GIn, Ala, Thr, Ser, and GIy;
  • R 2 is selected from the group consisting of NH 2 , and GIy-OH; providing that the compound has an isoelectric point in the range from about 6.0 to about 9.0 and further providing that when R 1 is His, X is Ala, Y is GIu, and Z is GIu, R 2 must be
  • the free compounds of the invention comprise a 1 ,2 aminothiol moiety, that is a moiety of the formula (IV):
  • R is a chemical group, preferably C 1 -C 6 alkyl or hydrogen, or the remainder of the molecule.
  • the remainder of the molecule may alternatively be attached at either carbon atom of the 1 ,2-aminothiol moiety.
  • the 1 ,2-aminothiol moiety is a cysteine residue.
  • Cysteine has the formula (V)
  • the remainder of the molecule may be attached at any chemically feasible point of the cysteine residue, for example:
  • the cysteine residue is suitably of either enantiomeric form. Preferably, it is of the naturally occurring (L) form.
  • the compound has the formula (X)
  • Re represents the remainder of the molecule, and R represents H or C 1 -C 6 alkyl, preferably H.
  • the optional linker group when present, forms a covalent link to both the moiety having GLP-1 like activity and the 1,2 aminothiol moiety.
  • R', R1 and R2 are independently selected from H and CrC 6 alkyl, and n is 0, 1 or 2, G is the moiety having GLP-1 like activity and At is the 1,2 aminothiol moiety;
  • polyatom linkers such as alkylene groups, polyethylene glycol (PEG) groups, diacids, diamines, aminoacids (especially ⁇ , ⁇ , ⁇ , and ⁇ ), aminoalcohols, diols, poly(aminoacids), bissulfides.
  • a preferred class of linker is aminoacids. Aminoacids of any length ( ⁇ , ⁇ , ⁇ , ⁇ and longer) may be employed. In one embodiment, the aminoacid is an ⁇ aminoacid. Preferably, the ⁇ aminoacid is a naturally occurring ⁇ aminoacid.
  • the aminoacid is a ⁇ aminoacid.
  • the aminoacid is 4-aminobutyric acid; i.e. the free compound of the invention has the structure (XV) or (XVI);
  • the free compound of the invention has the structure (XV).
  • the linker is a ⁇ aminoacid and the 1,2-aminothiol is a cysteine residue.
  • the free molecule of the invention has the formula (XVII)
  • the free molecule of the invention has the formula (XVIII)
  • linker (or the 1 ,2-aminothiol moiety if no linker is present) is attached to the moiety having GLP-1 like activity at any chemically appropriate site.
  • the 01489 is attached to the moiety having GLP-1 like activity at any chemically appropriate site.
  • the linker (or the 1 ,2-aminothiol moiety if no linker is present) is attached to an amine group present in the moiety having GLP-1 like activity.
  • the moiety having GLP-1 like activity comprises a lysine residue, preferably the linker is attached to the ⁇ -nitrogen of the lysine.
  • the point of attachment of the linker is the ⁇ -nitrogen of lysine 26.
  • the moiety having GLP-1 like activity is GLP-1 (7-37), and the point of attachment of the linker (or the 1 ,2-aminothiol moiety if no linker is present) is the ⁇ -nitrogen of lysine 26.
  • the free compound of the invention has the structure (XIX)
  • the moiety having GLP-1 like activity is GLP-1 (7-37), the linker is 4-aminobutyric acid and the point of attachment of the linker is the ⁇ -nitrogen of lysine 26.
  • the free compound of the invention has the structure (XX) moiety)
  • the moiety having GLP-1 like activity is GLP-1 (7-37), the 1,2- aminothiol moiety is cysteine, and the point of attachment of the linker is the ⁇ - nitrogen of lysine 26.
  • the free compound of the invention has the structure (XXI)
  • the free compounds of the invention preferably have insulinotropic activity.
  • insulinotropic activity refers to the property of a compound to stimulate the synthesis or expression of the hormone insulin.
  • the free compounds of the invention have insulinotropic activity that is at least 10 % of that of GLP-1 (7-36). More preferably, the free compounds of the invention have insulinotropic activity that is at least 20 % of that of GLP-1 (7-36). More preferably, the free compounds of the invention have insulinotropic activity that is at least 50 % of that of GLP-1 (7-36). More preferably, the free compounds of the invention have insulinotropic activity that is at least 80 % of that of GLP-1 (7-36). More preferably, the free compounds of the invention have insulinotropic activity that is at least equal to that of GLP-1 (7-36).
  • Insulinotropic activity in this context may be measured using a suitable assay.
  • the insulinotropic property of a compound may be determined by providing that compound to animal cells, or injecting that compound into animals and monitoring the release of immunoreactive insulin (IRI) into the media or circulatory system of the animal, respectively.
  • IRI immunoreactive insulin
  • the presence of IRI is detected through the use of a radioimmunoassay which can specifically detect insulin.
  • any radioimmunoassay capable of detecting the presence of IRI may be employed, it is preferable to use a modification of the assay method of Albano, J. D. M., et al., (Acta Endocrinol. 70, 487-509 (1972)).
  • a phosphate/albumin buffer with a pH of 7.4 was employed.
  • the incubation was prepared with the consecutive addition of 500 ⁇ l of phosphate buffer, 50 ⁇ l of perfusate sample or rat insulin standard in perfusate, 100 ⁇ l of anti-insulin antiserum (Wellcome Laboratories; 1 :40,000 dilution), and 100 ⁇ l of [ 125 I] insulin, giving a total volume of 750 ⁇ l in a 10*75-mm disposable glass tube.
  • the assay sensitivity was 1-2 ⁇ U/ml.
  • radioactive label In order to measure the release of IRl into the cell culture medium of cells grown in tissue culture, one preferably incorporates radioactive label into proinsulin.
  • any radioactive label capable of labelling a polypeptide can be used, it is preferable to use 3 H leucine in order to obtain labelling of proinsulin. Labeling can be done for any period of time sufficient to permit the formation of a detectably labelled pool of proinsulin molecules; however, it is preferable to incubate cells in the presence of radioactive label for a 60-minute time period.
  • any cell line capable of expressing insulin can be used for determining whether a compound has an insulinotropic effect, it is preferable to use rat insulinoma cells, and especially RIN-38 rat insulinoma cells.
  • Such cells can be grown in any suitable medium; however, it is preferable to use DME medium containing 0.1% BSA and 25 mM glucose.
  • the insuiinotropic property of a compound may also be determined by pancreatic infusion.
  • the in situ isolated perfused rat pancreas preparation was a modification of the method of Penhos, J. C 1 et al. (Diabetes 18, 733-738 (1969)).
  • fasted rats preferably male Charles River strain albino rats
  • weighing 350-600 g are anesthetized with an intraperitoneal injection of Amytal Sodium (EIi Lilly and Co., 160 ng/kg). Renal, adrenal, gastric, and lower colonic blood vessels are ligated.
  • the entire intestine is resected except for about four cm of duodenum and the descending colon and rectum. Therefore, only a small part of the intestine is perfused, thus minimizing possible interference by enteric substances with glucagon-like immunoreactivity.
  • the perfusate is preferably a modified Krebs- Ringer bicarbonate buffer with 4% dextran T70 and 0.2% bovine serum albumin (fraction V), and is preferably bubbled with 95% O 2 and 5% CO 2 .
  • a nonpulsatile flow, four-channel roller-bearing pump (Buchler polystatic, Buchler Instruments Division, Nuclear-Chicago Corp.) is preferably used, and a switch from one perfusate source to another is preferably accomplished by switching a three-way stopcock.
  • the manner in which perfusion is performed, modified, and analyzed preferably follows the methods of Weir, G. C, et al., (J. Clin. Investigat. 54:1403-1412 (1974)), which is hereby incorporated by reference.
  • the free compounds of the invention show increased resistance to deactivation by DPP IV compared with natural GLP-1.
  • Resistance to DPP IV deactivation is suitably measured for example by the technique described in in Green, B.D. et al, Biological Chemistry, 385, 169-177,(2004).
  • Type 2 diabetes therapies may depend on their incorporation into suitable extended release formulations.
  • Such free compounds may block elimination through the urinary pathway and confer a degree of proteolytic protection on the attached peptide, thus prolonging duration of action.
  • Preferred moieties having GLP-1 like activity include those of the formula (XXII)
  • X is optionally present and represents a substituent of the terminal carboxyl or amino group
  • Y is a linker group
  • n is an integer
  • n 1 or 2;
  • each A is an independently selected amino acid
  • each B is an independently selected amino acid which may be further optionally modified and/ or substituted.
  • Z is optionally present and represents a substituent of the terminal amino or carboxy group; provided that the compound is not GLP-1 or a naturally occurring fragment thereof;
  • the moiety having GLP-1 like activity of the invention has the formula (XXIIl)
  • R 1 and R 2 represent the backbone/sidechain of the amino acids
  • P is H, a substituent, or a link to R 1 or R 2 (e.g. in the case of proline)
  • K represents H, H 2 , NH or O.
  • the moiety having GLP-1 like activity of the invention has the formula (XXIV)
  • Amino acids Ai and A 2 may be ⁇ , ⁇ , ⁇ or other amino acids.
  • Ai and A 2 are ⁇ amino acids.
  • amino acids Ai and A 2 are independently selected from groups of the formula (XXV)
  • R 3 and R 4 are independently selected from hydrogen atom or an alkyl, aminoalkyl (optionally substituted on the nitrogen atom by one or two alkyl, phenyl, benzyl, cycloalkyl, optionally substituted aryloxycarbonyl, optionally substituted arylalkoxycarbonyl and/or optionally substituted alkoxycarbonyl groups), thioalkyl (optionally substituted on the sulphur atom by an alkyl, phenyl, benzyl or cycloalkyl group), hydroxyalkyl (optionally substituted on the oxygen atom by an alkyl, phenyl, benzyl or cycloalkyl group), carboxyalkyl, carbamoylalkyl, guanidinoalkyl, cycloalkyl, cycloalkylalkyl, optionally substituted fused cycloalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally
  • P is selected from hydrogen, alkyl, or cycloalkyl, or taken with R 2 or R 3 together with the carbon and nitrogen atoms to which they are attached represents a mono- or bicyclic group having from 4 to 12 ring members which is saturated, partially unsaturated, or unsaturated and is optionally substituted.
  • At least one of R 3 and R 4 is hydrogen.
  • the amino acids A 1 and A 2 may have the D- or L- configuration.
  • the amino acids Ai and A 2 have the L- configuration.
  • the amino acids Ai and A 2 have the D- configuration.
  • a 1 is His. More preferably, Ai is D-His.
  • a 2 is Ala. More preferably, A 2 is D-AIa. More preferably, Ai is His and A 2 is Ala. More preferably, Ai is D-His and A 2 is D-AIa.
  • the moiety having GLP-1 like activity has the formula (XXIV), A 1 is D-His and A 2 is D-AIa; that is the moiety having GLP-1 like activity has the formula (XXVI)
  • X, Y, B 1 n and Z have the meanings defined above. Still more preferably, the N-terminus of the peptide [B] n is bound covalently to the group Y.
  • Group X is preferably selected from a hydrogen atom, alkyl, alkenyl, alkynyl, acyl, alkoxy, cycloalkyl, cycloalkoxy, thioalkyl, alkylthio, sulfoxoalkyl, haloalkyl, aryl, heteroaryl, NH 2 , NH(Alkyl), N(Alkyl) 2 , alkyleneNH 2 , AlkyleneNHAIkyl, Alkylene(Alkyl) 2 , NHAryl, N(Aryl) 2 and heteroaryl.
  • group X has basic character.
  • Basic character in this context means that group X is capable of binding a proton in aqueous solution at pH 7.
  • Highly preferred groups X are 6)alkyl 2 (alkyl the same or different), -NH(C- ⁇ -6 )alkyl, -N(C 1-6 )alkyl 2 , and -NH 2 . Most preferred is -NH 2 .
  • the moiety having GLP-1 like activity of the invention has the formula (XXV) above and X is -NH 2 ; that is the compound has the formula (XXVII)
  • Y, B, n and Z are as defined above.
  • Each group B is an independently selected amino acid.
  • the groups B are linked together in a conventional manner (i.e. via amide bonds) to form a peptide chain.
  • the groups B are numbered according to formula (XXVIII);
  • the N-terminus of the group [B] n is covalently bound to the group Y, and the C-terminus of the group [B] n is covalently bound to the group Z where present.
  • the C-terminus of the group [B] n is covalently bound to the group Y, and the N-terminus of the group [B] n is covalently bound to the group Z where present.
  • Amino acids B may be ⁇ , ⁇ , ⁇ or other amino acids.
  • B are independently selected ⁇ amino acids.
  • the amino acids B may have the D- or L- configuration.
  • amino acids B are independently selected from groups of the formula (XXIX)
  • R 5 and R 6 are independently selected from hydrogen atom or an alkyl, aminoalkyl (optionally substituted on the nitrogen atom by one or two alkyl, phenyl, benzyl, cycloalkyl, optionally substituted aryloxycarbonyl, optionally substituted arylalkoxycarbonyl and/or optionally substituted alkoxycarbonyl groups), thioalkyl (optionally substituted on the sulphur atom by an alkyl, phenyl, benzyl or cycloalkyl group), hydroxyalkyl (optionally substituted on the oxygen atom by an alkyl, phenyl, benzyl or cycloalkyl group), carboxyalkyl, carbamoylalkyl, guanidinoalkyl, cycloalkyl, cycloalkylalkyl, optionally substituted fused cycloalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally
  • Q is selected from hydrogen, alkyl, or cycloalkyl, or taken with R 5 or R 6 together with the carbon and nitrogen atoms to which they are attached represents a mono- or bicyclic group having from 4 to 12 ring members which is saturated, partially unsaturated, or unsaturated and is optionally substituted.
  • At least one of R 5 and R 6 is hydrogen.
  • amino acids B are independently selected from naturally occurring amino acids.
  • n is less than 100. More preferably, n is less than 50. More preferably, n is less than 30. More preferably, n is 28 or 29.
  • the amino acid sequence [B] n is substantially homologous to GLP-1 (9- 37) or a fragment of GLP-1 having n residues.
  • substantially homologous means that at least 80 %, preferably at least 90 % and more preferably at least 95 % of the amino acid sequence is the same.
  • GLP-1 (1 to 37) has the sequence His Asp GIu Phe GIu Arg His Ala GIu GIy Thr Phe Thr Ser Asp VaI Ser Ser Tyr Leu GIu GIy GIn Ala Ala Lys GIu Phe lie Ala Trp Leu VaI Lys GIy Arg GIy .
  • GLP-1 (7 to 37) has the sequence His Ala GIu GIy Thr Phe Thr Ser Asp VaI Ser Ser Tyr Leu GIu GIy GIn Ala Ala Lys GIu Phe He Ala Trp Leu VaI Lys GIy Arg GIy .
  • GLP-1 (9 to 37) has the sequence GIu GIy Thr Phe Thr Ser Asp VaI Ser Ser Tyr Leu GIu GIy GIn Ala Ala Lys GIu Phe lie Ala Trp Leu VaI Lys GIy Arg GIy .
  • B n is selected from a group of the formula
  • each of B 1 , B 7 , B 8 , B 10 , B 11 , B 12 , B 17 , B 19 , B 20 , B 21 , B 23 , B 24 and B 28 is independently selected from the definition of B above.
  • B n is selected from a group of the formula
  • 0 , B 11 , B 12 , B 17 , Bi 9 , B 2D , B 21 , B 23 , B 24 and B 2B is independently selected from the definition of B above.
  • B n is selected from a group having the formula
  • [B] n comprises a sequence of the formula
  • [B] n has the formula
  • linker group Y is defined as a chemical moiety capable of forming a covalent bond with the C or N terminus of [B] n and the C or N terminus of A 1 -A 2 .
  • Preferred linker groups Y are those of the formula (XXX)
  • ⁇ represents an optional single or double bond
  • the group Y can also be a bond (single, double or triple).
  • the group Y may independently feature single, double or triple bonds to amino acids Ai or A 2 and Bi.
  • the carboxy and/or amino termini linked to Y may be in any oxidation state.
  • R 7 is selected from O, NH 1 N(C 1-6 )Alkyl, and S.
  • R 7 is O.
  • the group Y has the formula (XXXII)
  • X' is selected from O, NH or N(C1-C6)alkyl
  • Y' is selected from O, NH or N(C1-C6)alkyl
  • Rg and R 10 are independently selected from hydrogen atom or an alkyl, aminoalkyl (optionally substituted on the nitrogen atom by one or two alkyl, phenyl, benzyl, cycloalkyl, optionally substituted aryloxycarbonyl, optionally substituted arylalkoxycarbonyl and/or optionally substituted alkoxycarbonyl groups), thioalkyl (optionally substituted on the sulphur atom by an alkyl, phenyl, benzyl or cycloalkyl group), hydroxyalkyl (optionally substituted on the oxygen atom by an alkyl, phenyl, benzyl or cycloalkyl group), carboxyalkyl, carbamoylalkyl, guanidinoalkyl, cycloalkyl, cycloalkylalkyl, optionally substituted fused cycloalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally
  • Y' is NH or O.
  • Rg is H or methyl.
  • R- J0 is H or methyl.
  • R 9 is H and Ri 0 is methyl, or R 9 is H and Ri 0 is methyl. Alternatively, and preferably, R 9 and Ri 0 are H.
  • group Y has the formula (XXXIII, XXXIV or XXXV)
  • the compound has the formula (XXXVI)
  • R 8 is selected from O, S 1 NH 1 and N(Alkyl).
  • R 8 is O.
  • the compounds of the invention feature a urea group linking A2 and [B] n .
  • the compound has the formula (XXXVIII)
  • R 2 represents the backbone/sidechain of the amino acids
  • P is H, a substituent, or a link to R 1 or R 2 (e.g. in the case of proline)
  • _. represents an optional single or double bond
  • K represents H, H 2 , NH or O.
  • Group Z 1 where present, is preferably selected from a hydrogen atom, alkyl, alkenyl, alkynyl, acyl, alkoxy, cycloalkyl, cycloalkoxy, thioalkyl, alkylthio, sulfoxoalkyl, haloalkyl, aryl, heteroaryl, NH 2 , NH(Alkyl), and N(Alkyl) 2 .
  • the compound has the formula
  • the invention relates to a free compound of the invention covalently linked to a carrier iigand ("complexed compound").
  • the carrier ligand is linked to the free compound via the aminothiol moiety.
  • the carrier ligand in its free state (i.e. prior to attachment to the free compound) has a carbonyl group which serves as the point of attachment of the free compound.
  • the 1 ,2 aminothiol moiety forms a thiazolidine group with the carbonyl group of the carrier ligand, as shown in scheme 1.
  • Re represents the remainder of the molecule
  • R represents chemical group, preferably C 1 -C 6 alky! or hydrogen
  • X and Y represent chemical groups.
  • the carrier ligand in its free state is polymeric or oligomeric.
  • the complexed compound has a longer half life in vivo compared with the free compound.
  • the complexed compound preferably has intrinsic activity, that is it has GLP-1 like activity; however, the complexed compound may lack intrinsic activity, and only be transformed to an active compound (i.e. one possessing GLP-1 like activity) when in vivo. In this latter aspect, the complexed molecule behaves as a pro-drug.
  • the carrier ligand is a poly- or oligosaccharide.
  • the polysaccharide compound may be a naturally occurring poly- or oligosaccharide, a derivative of a naturally occurring poly- or oligosaccharide, for instance a poly- or oligosaccharide which has been derivatised by reaction of one or more active groups on the saccharide residues, or which has been covalently linked to a derivatising group by either end of the poly- or oligosaccharide chain or by an active group midway along the chain, or may be a naturally occurring poly- or oligosaccharide derivative, for instance comprising attached phospholipids or proteins, or derivatives of naturally occurring poly- or oligosaccharide derivatives, for instance chemically derivatised compounds, such as hydrolysed or otherwise chemically reacted derivatives.
  • the poly- or oligosaccharide portion of the compound has more than 5, preferably at least 10, and more preferably at least 20 or 50 monosaccharide residues in the polymer chain.
  • Readily available polysaccharide compounds may have up to 500 saccharide residues in total, but usually have fewer than 300 residues in the polymer chain.
  • the ligand is a polyether moiety.
  • polyether moiety refers to a group containing a plurality of ether linkages.
  • a highly preferred polyether ligand is a PEG (polyethylene glycol) iigand.
  • PEG polyethylene glycol
  • ligands comprise at least a repeat ethylene glycol unit, , wherein n is an integer.
  • PEG groups are known and described in the art.
  • the term PEG includes poly(ethylene glycol) in any of its forms, including alkoxy PEG, difunctional PEG, multiarmed PEG, forked PEG, branched PEG, pendent PEG (i.e. PEG or related polymers having one or more functional groups pendent to the polymer backbone), or PEG with degradable linkages therein.
  • the polymer backbone can be linear or branched. Branched polymer backbones are generally known in the art.
  • a branched polymer has a central branch core moiety and a plurality of linear polymer chains linked to the central branch core.
  • PEG is commonly used in branched forms that can be prepared by addition of ethylene oxide to various polyols, such as glycerol, pentaerythritol and sorbitol.
  • the central branch moiety can also be derived from several amino acids, such as lysine.
  • the branched poly(ethylene glycol) can be represented in general form as R(-PEG-OH).sub.m in which R represents the core moiety, such as glycerol or pentaerythritol, and m represents the number of arms.
  • Multi-armed PEG molecules such as those described in U.S. Pat. No. 5,932,462, which is incorporated by reference herein in its entirety, can also be used as the polymer backbone.
  • the ligand is carbonyl-containing polyether moiety; that is, a moiety comprising at least one polyether group and at least one carbonyl group.
  • the carbonyl group is an aldehyde group.
  • the ligand is an aldehyde-containing poly(ethyleneglycol) moiety (PEG- CHO). More preferably, the ligand is of the formula (LXVIII)
  • R a is a C 1 -C 6 alkyl group
  • AIk is a branched or straight C 1 -C 6 alkyiene group.
  • the complexed compound of the invention preferably has the formula LXIX, as shown in scheme 4.
  • R a , Re, R, AIk and n have the meanings set out above.
  • the compound of the invention has the formula LXX
  • the complexed compounds of the invention have intrinsic GLP-1 like activity. Alternatively, the complexed compounds of the invention do not have intrinsic activity, and behave as pro-drugs of the activity species.
  • the invention also encompasses a process for the preparation of the compounds of formula (I) which may be obtained by various methods.
  • Convenient methods include solid phase sequential synthesis, synthesis and coupling of fragments in solution, enzymatic synthesis, or by employing molecular biology techniques.
  • Solid phase synthesis is for example carried out using an automatic device which executes in a repetitive and programmable manner the deprotection, coupling and washing cycles necessary for the sequential introduction of amino acids into the peptide chain.
  • the C-terminal amino acid is fixed on a resin conventionally used for the preparation of polypeptides, preferably a polystyrene cross-linked with 0.5 to 3.0% divinylbenzene and provided with activated radicals that enable the first amino acid to be fixed covalently to the resin.
  • a resin conventionally used for the preparation of polypeptides preferably a polystyrene cross-linked with 0.5 to 3.0% divinylbenzene and provided with activated radicals that enable the first amino acid to be fixed covalently to the resin.
  • Suitable resins are well known to those skilled in the art. Appropriate selection of the resin allows the introduction after synthesis of the C- terminal Z function.
  • the amino acids are then introduced one by one in the desired sequence.
  • Each cycle of synthesis corresponding to the introduction of an amino acid comprises N- terminal deprotection of the peptide chain, successive washings designed to remove the reagents or swell the resin, coupling with activation of the amino acid and further washings.
  • Each of those operations is followed by filtration effected as a result of the presence of a sintered glass filter incorporated in the reactor in which the synthesis is beings carried out.
  • the couplings reagents used are the conventional reagents for peptide synthesis, such as dicyclohexylcarbodiimide (DCC) and hydroxybe ⁇ zotriazole (HOBT) or benzotriazol-1-yl-oxytris(dimethylarnino)phosphonium hexafluorophosphate (BOP), or also diphenyl-phosphorylazide (DPPA). Activation by the formation of mixed or symmetrical anhydrides is also possible.
  • DCC dicyclohexylcarbodiimide
  • HOBT hydroxybe ⁇ zotriazole
  • BOP benzotriazol-1-yl-oxytris(dimethylarnino)phosphonium hexafluorophosphate
  • DPPA diphenyl-phosphorylazide
  • Each amino acid is introduced into the reactor in an approximately 6-fold excess in relation to the degree of substitution of the resin and in an approximately equivalent amount in relation to the coupling agents.
  • the coupling reaction may be confirmed at each stage of the synthesis by the ninhydrin reaction test described by Kaiser et al. (Anal. Biochem., 34, 595, 1970).
  • an appropriate treatment for example using a strong acid such as trifluoroacetic acid, or hydrofluoric acid in the presence of anisole, ethanedithiol or 2-methylindole, is used to cleave the peptide from the resin and also to free the peptide of its protecting groups.
  • a strong acid such as trifluoroacetic acid, or hydrofluoric acid in the presence of anisole, ethanedithiol or 2-methylindole
  • the group Z may be introduced at this stage, concomitant with cleavage from the resin.
  • the skilled person will be able to select suitable conditions for such a transformation.
  • the peptide may be cleaved from the resin with a free carboxyl group, which may subsequently be coupled to the group Z.
  • the skilled person will be able to determine suitable conditions.
  • the compound is then optionally purified by conventional purification techniques, especially chromatography techniques.
  • the 1 ,2 aminothiol moiety and linker group may be introduced at any appropriate stage of peptide synthesis.
  • the linker group is a 4-amino butyric acid group
  • the 1 ,2 aminothiol moiety is a cysteine residue
  • the moiety having GLP-1 like activity is GLP- 1 (7-37)
  • the point of attachment is Lys 26
  • the free compound of the invention is suitably synthesised according to scheme 5.
  • the peptide may be assembled in the N to C direction. Suitable methodology is described in GB0505200.6, the contents of which are incorporated herein by reference.
  • the peptides of the present invention may also be obtained by coupling in solution selectively protected peptide fragments which may themselves be prepared either in the solid phase or in solution.
  • the use of protecting groups and the exploitation of their differences in stability is analogous to solid phase methods with the exception of the attachment of the peptide chain to the resin.
  • the C-terminal carboxy group is protected, for example, by a methyl ester or an amide function.
  • the methods of activation during coupling are likewise analogous to those employed in solid phase synthesis.
  • the peptides of the present invention may also be obtained using molecular biology techniques, employing nucleic acid sequences that encode those peptides. Those sequences may be RNA or DNA and may be associated with control sequences and/or inserted into vectors. The latter are then transfected into host cells, for example bacteria. The preparation of the vectors and their production or expression in a host are carried out by conventional molecular biology and genetic engineering techniques.
  • linker group Y may be achieved as a step in the solid-phase synthesis of peptides as described above, or alternatively in solution.
  • compounds of formula (XXVII) above wherein R 8 is O are suitably prepared as shown in scheme 6.
  • the invention also encompasses all pharmaceutically acceptable forms of compounds of formula I, including without limitation, its free form (zwitterion), and its pharmaceutically acceptable complexes, salts, solvates, hydrates, and polymorphs.
  • Salts include, without limitation, acid addition salts and base addition salts, including hemisalts.
  • the present invention also encompasses pharmaceutically acceptable salts of the present compounds.
  • Such salts include pharmaceutically acceptable acid addition salts, pharmaceutically acceptable metal salts, ammonium and alkylated ammonium salts.
  • Acid addition salts include salts of inorganic acids as well as organic acids.
  • suitable inorganic acids include hydrochloric, hydrobromic, hydriodic, phosphoric, sulfuric, nitric acids and the like.
  • suitable organic acids include formic, acetic, trichloroacetic, trifluoroacetic, propionic, benzoic, cinnamic, citric, fumaric, glycolic, lactic, maleic, malic, malonic, mandelic, oxalic, picric, pyruvic, salicylic, succinic, methane-sulfonic, ethanesulfonic, tartaric, ascorbic, pamoic, bismethylene salicylic, ethanedisulfonic, gluconic, citraconic, aspartic, stearic, palmitic, EDTA, glycolic, p- aminobenzoic, glutamic, benzenesulfonic, p-toluenesulfonic acids and the like.
  • metal salts include lithium, sodium, potassium, magnesium, calcium salts and the like.
  • ammonium and alkylated ammonium salts include ammonium, methyl-, dimethyl-, trimethyl-, ethyl-, hydroxyethyl-, diethyl-, n-butyl-, sec- butyl-, tert-butyl-, tetramethylammonium salts and the like.
  • a compound according to the invention is used in the treatment or prevention of a condition selected from hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, atheroschlerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcers.
  • a condition selected from hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, atheroschlerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcers.
  • the compounds of the present invention may be combined in use with one or more pharmacologically active substances.
  • Preferred classes of pharmacologically active substances are antidiabetic agents, antiobesity agents, appetite regulating agents, antihypertensive agents, agents for the treatment and/or prevention of complications resulting from or associated with diabetes and agents for the treatment and/or prevention of complications and disorders resulting from or associated with obesity.
  • insulin resistance is the pathophysiological mechanism.
  • Examples of these pharmacologically active substances are: Insulin, GLP-1 agonists, sulphonylureas (e. g. tolbutamide, glibenclamide, glipizide and gliclazide), biguanides e. g. metformin, meglitinides, glucosidase inhibitors (e. g.
  • acorbose glucagon antagonists
  • DPP-IV dipeptidylpeptidase-IV
  • inhibitors of hepatic enzymes involved in stimulation of gluconeogenesis and/or glycogenosis glucose uptake modulators
  • thiazolidinediones such as troglitazone and ciglitazone
  • compounds modifying the lipid metabolism such as antihyperlipidemic agents as HMG CoA inhibitors (statins), compounds lowering food intake, RXR agonists and agents acting on the ATP-dependent potassium channel of the micelles, e. g.
  • glibenclamide glipizide, gliclazide and repaglinide
  • Cholestyramine colestipol, clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin, probucol, dextrothyroxine, neteglinide, repaglinide
  • ⁇ -blockers such as alprenolol, atenolol, timolol, pindolol, propranolol and metoprolol
  • ACE angiotensin converting enzyme
  • benazepril capto pril, enalapril, fosinopril, lisinopril, alatriopril, quinapril and ramipril, calcium channel .
  • ⁇ -blockers such as nifedipine, felodipine, nicardipine, isradipine, nimodipine, diltiazem and verapamil, and ⁇ -blockers such as doxazosin, urapidil, prazosin and terazosin; CART (cocaine am-phetamine regulated transcript) agonists, NPY (neuropeptide Y) antagonists, MC4 (melanocortin 4) agonists, orexin antagonists, TNF (tumor necrosis factor) agonists, CRF (corticotropin releasing factor) agonists, CRF BP (corticotropin releasing factor binding protein) antagonists, urocortin agonists, ss3 agonists, MSH (melanocyte-stimulating hormone) agonists, MCH (melanocyte-concentrating hormone) antagonists, CCK (cholecystokinin) agonists, serotonin
  • the compound of the invention and the additional therapeutic agent may be present in the same dosage form, or separately for sequential, separate or simultaneous administration.
  • the present invention also provides pharmaceutical compositions comprising a compound of the present invention in combination with a pharmaceutically acceptable carrier, diluent, or excipient.
  • Such pharmaceutical compositions are prepared in a manner well known in the pharmaceutical art, and are administered individually or in combination with other therapeutic agents, preferably via parenteral routes.
  • Especially preferred routes include intramuscular and subcutaneous administration.
  • Parenteral daily dosages are in the range from about 1 pg/kg to about 1 ,000 ⁇ g/kg of body weight, although lower or higher dosages may be administered.
  • the required dosage will depend upon the severity of the condition of the patient and upon such criteria as the patient's height, weight, sex, age, and medical history.
  • the active ingredient which comprises at least one compound of the present invention, is usually mixed with an excipient or diluted by an excipient.
  • an excipient When an excipient is used as a diluent, it may be a solid, semi-solid, or liquid material which acts as a vehicle, carrier, or medium for the active ingredient.
  • the active compound In preparing a formulation, it may be necessary to mill the active compound to provide the appropriate particle size prior to combining with the other ingredients. If the active compound is substantially insoluble, it ordinarily is milled to particle size of less than about 200 mesh. If the active compound is substantially water soluble, the particle size is normally adjusted by milling to provide a substantially uniform distribution in the formulation, e.g., about 40 mesh.
  • compositions of the invention can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the patient by employing procedures well known in the art.
  • compositions are preferably formulated in a unit dosage form with each dosage normally containing from about 50 ⁇ g to about 100 mg, more usually from about 1 mg to about 10 mg of the active ingredient.
  • unit dosage form refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with a suitable pharmaceutical excipient.
  • compositions according to the invention there are those which are suitable for oral, parenteral or nasal administration, including tablets or dragees, sublingual tablets, sachets, soft gelatin capsules, suppositories, creams, ointments, dermal gels, transdermal devices, aerosols, drinkable and injectable ampoules.
  • compositions containing a compound of the present invention preferably are combined with distilled water and the pH is adjusted to about 6.0 to about 9.0.
  • Controlled release preparations may be achieved by the use of polymers to complex or absorb a compound of the present invention.
  • the controlled delivery may be exercised by selecting appropriate macromolecules (for example, polyesters, polyamino acids, polyvinylpyrrolidone, ethylenevinyl acetate, methylcellulose, carboxymethylcellulose, and protamine sulfate) and the concentration of macromolecules as well as the methods of incorporation in order to control release.
  • Another possible method to control the duration of action by controlled release preparations is to incorporate a compound of the present invention into particles of a polymeric material such as polyesters, polyamino acids, hydrogels, poly (lactic acid) or ethylene vinylacetate copolymers.
  • a polymeric material such as polyesters, polyamino acids, hydrogels, poly (lactic acid) or ethylene vinylacetate copolymers.
  • microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules, respectively, or in colloidal drug delivery systems, for example, liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules, or in macroemulsions.
  • colloidal drug delivery systems for example, liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules, or in macroemulsions.
  • the compounds of the present invention have insulinotropic activity.
  • another aspect of the present invention provides a method for enhancing the expression of insulin comprising providing to a mammalian pancreatic B-type islet cell an effective amount of a compound of the present invention.
  • Also intended as pharmaceutically acceptable acid addition salts are the hydrates and other solvates which the present compounds are able to form.
  • the pharmaceutically acceptable salts comprise basic amino acid salts such as lysine, arginine and ornithine.
  • the acid addition salts may be obtained as the direct products of compound synthesis.
  • the free base may be dissolved in a suitable solvent containing the appropriate acid, and the salt isolated by evaporating the solvent or otherwise separating the salt and solvent,
  • the compounds of the present invention may form solvates with standard low molecular weight solvents using methods well known to the person skilled in the art. Such solvates are also contemplated as being within the scope of the present invention.
  • the invention also encompasses prodrugs of the present compounds, which on administration undergo chemical conversion by metabolic processes before becoming pharmacologically active substances.
  • prodrugs will be functional derivatives of the compounds of the general formula (I), which are readily convertible in vivo into the required compound of the formula (I).
  • Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", ed. H. Bundgaard, Elsevier, 1985.
  • GLP-1 For example, it is well known that several forms of GLP-1 are processed in vivo from proglucagon; GLP-1 (1-37), GLP-1 (7-37) and GLP-1 (7-36). Accordingly, the person skilled in the art will understand that the present invention also embraces peptides incorporating a fragment of structure (I) above which are metabolisable in vivo to give compounds of formula (I).
  • the invention also encompasses active metabolites of the present compounds.
  • N(alkyl) 2 refers to an amine group having two independently selected alkyl substituents.
  • a group or substituent is itself defined as optionally substituted, unless otherwise specified, it has up to three substituents independently selected from the group consisting of halogen, OH, NH 2 , NH(alkyl), N(alkyl) 2 , alkyl, alkenyl, alkynyl, alkoxy, carbomoyl, carboxy, cyano, nitro, and SH.
  • substituents independently selected from the group consisting of halogen, OH, NH 2 , NH(alkyl), N(alkyl) 2 , alkyl, alkenyl, alkynyl, alkoxy, carbomoyl, carboxy, cyano, nitro, and SH.
  • Aikyl refers to an aliphatic hydrocarbon chain and includes straight and branched chains preferably of 1 to 12, more preferably 1 to 6 carbon atoms such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, neo-pentyl, n-hexyl, and isohexyl.
  • alkylene refers to a divalent saturated branched or unbranched hydrocarbon chain containing from 1 to 6 carbon atoms, and includes, for example, methylene (CH 2 ), ethylene (CH 2 CH 2 ), propylene (CH 2 CH 2 CH 2 ), 2- methylpropylene (CH 2 CH(CH 3 )CH 2 ), hexylene ((CH 2 ) 6 ), and the like.
  • Alkenyl refers to an aliphatic hydrocarbon chain having at least one double bond, and preferably one double bond, and includes straight and branched chains e. g. of 2 to 6 carbon atoms such as ethenyl, propenyl, isopropenyl,but-1-enyl, but-2-enyl, but-3-enyl, 2-methypropenyl.
  • Alkynyl refers to an aliphatic hydrocarbon chain having at least one triple bond, and preferably one triple bond, and includes straight and branched chains e. g. of 2 to 6 carbon atoms such as ethynyl, propynyl, but-1-ynyl, but-2-ynyl and but-3-ynyl.
  • Cycioalkyl refers to a cyclic, saturated hydrocarbon group having from 3 to 8 ring carbon atoms.
  • Examples of cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.
  • Cycloalkylene refers to a divalent saturated cyclic hydrocarbon group containing from 3 to 8 ring carbon atoms, and includes, for example, methylene cyclopropylene, cyclobutylene, cyclohexylene and the like.
  • cycloalkylalkyl refers to the group -alkylene-cycloalkyl, wherein alkylene and cycloalkyl are as defined above.
  • Examples of cycloalkylalkyl . groups include cyclopropylmethyl, 2-cyclobutylethyl, cyclopentylmethyl, cyclohexylmethyl and 4-cycloheptylbutyl, and the like.
  • Alkoxy as used herein refers to the group -O-alkyl, wherein alkyl is as defined above.
  • alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n- butoxy, isobutoxy, sec-butoxy, t-butoxy, n-pentoxy, isopentoxy, neo-pentoxy, n- hexyloxy, and isohexyloxy.
  • AIkylthio refers to the group -S-alkyl, wherein alkyl is as defined above.
  • alkylthio groups include thiomethyl, thioethyl and thiohexyl.
  • Cycloalkoxy as used herein refers to the group -O-cycloalkyl, wherein cycloalkyl is as defined above.
  • Examples of cycloalkoxy groups are cyclopropoxy, cyclobutoxy, cyclopentoxy, cyclohexyloxy, cycloheptyloxy and cyclooctyloxy.
  • Aminoalkyl as used herein refers to the term -alkylene-NH 2 , wherein alkylene is as defined above.
  • Examples of aminoalkyl groups include methylamino (-CH 2 NH 2 ) and 2-ethylamino (-CH 2 CH 2 NH 2 ).
  • Thioalkyl as used herein refers to the group -alkylene-SH, wherein alkylene is as defined above.
  • Examples of thioalkyl groups are methylthio (CH 2 SH) and 2-ethylthio (CH 2 CH 2 SH). Sulfoxoalkyl
  • Sulfoxoalkyl refers to the group -S(O)-alkyl, wherein alkyl is as defined above.
  • Sulfonoalkyl refers to a the group -S(O) 2 -alkyl wherein alkyl is as defined above.
  • Halogen, halide or halo- refers to iodine, bromine, chlorine and fluorine.
  • Haloalkyl as used herein refers to an alkyl group as defined above wherein at least one hydrogen atom has been replaced with a halogen atom as defined above.
  • haloalkyl groups include chloromethyl, dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl and trifluoromethyl.
  • Preferred haloalkyl groups are fluoroalkyl groups (i.e. haloalkyl groups containing fluorine as the only halogen). More highly preferred haloalkyl groups are perfluoroalkyl groups, i.e. alkyl groups wherein all the hydrogen atoms are replaced with fluorine atoms.
  • Hydroxyalkyl refers to an alkyl group as defined above wherein at least one hydrogen atom is replaced by a hydroxyl group.
  • hydroxyalkyl groups include hydroxymethyl (-CH 2 OH), 2-hydroxyethyl (-CH 2 CH 2 OH), and 1- hydroxyethyl (-CH(OH)CH 3 ).
  • aryl refers to an unsaturated aromatic carbocyclic group of from 6 to 10 carbon atoms having a single ring (e. g., phenyl) or multiple condensed (fused) rings (e.g., naphthyl).
  • Preferred aryl groups include phenyl, naphthyl and the like.
  • Heteroaryl refers to 5 to 10 membered mono or bicyclic aromatic rings having from 1 to 3 heteroatoms selected from N, O and S.
  • Monocyclic rings preferably have 5 or 6 members and bicyclic rings preferably have 8, 9 or 10 membered ring structures.
  • Exemplary heteroaryls include pyrrolyl, furyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyrazinyl, pyrimidinyl, indolyl, quinolyl and isoquinolyl.
  • alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl and propoxycarbonyl.
  • aryloxycarbonyl groups include phenoxycarbonyl and naphthyloxycarbonyl.
  • carboxyalkyl refers to the group -alkylene-CO 2 H, wherein alkylene is as defined above.
  • carbamoyl refers to the group -CONH 2 .
  • carbamoylalkyl refers to the group -alkylene-carbamoyl, wherein alkylene and carbamoyl are as defined above.
  • Arylalkyl as used herein refers to the group -alkylene-aryl, wherein alkylene and aryl are as defined above.
  • Examples of arylalkyl groups are benzyl (-CH 2 Ph) and 2- phenethyl (-CH 2 CH 2 Ph).
  • Arylalkoxy as used herein refers to the group -O-alkylene-aryl, wherein alkylene and aryl are as defined above.
  • Examples of arylalkoxy groups include 2-phenylethoxy (- OCH 2 CH 2 Ph) 1 5-phenylpentyloxy (-O-(CH 2 ) 5 Ph) and the like.
  • heteroarylalkyl refers to the group -alkylene-heteroaryl, wherein alkylene and heteroaryl are as defined above.
  • imidazolylalkyl refers to the group -alkylene-imidazole, wherein alkylene is as defined above.
  • prodrugs that is compounds capable of undergoing metabolism to give compounds of formula (I) as defined above.
  • Suitable prodrugs are N-oxides and compounds having a quaternary nitrogen.
  • leaving group refers to any moiety or atom that is susceptible to nucleophilic substitution or elimination. Typically, these are atoms or moieties that when removed by nucleophilic substitution or elimination are stable in anionic form.
  • Examples of leaving groups useful in the present invention include alkyl- or arylsulphonate groups such as tosylate, brosylate, mesylate or nosylate, or halides such as fluoride, chloride, bromide, or iodide.
  • variable e.g. aryl, heterocycle, R 7 etc.
  • each peptide was dissolved in 0.5 ml of 50 mM triethanolamine buffer, pH 7.8. To this was added a 10 microlitre solution of 11 milliunits DPP-IV (Sigma) and the solutions incubated at 37 0 C for up to 42 hours. A control peptide, GLP-1 7-22, was treated identically with DPP-IV. The incubations were terminated by addition of 5 microlitres TFA to each tube and 50 microlitre of each were injected onto a reversed phase 4.6 x 250 mm Ci 8 HPLC column, eluting a gradient of 20% - 45% acetonitrile in 0.1% TFA/water over 25 minutes.
  • GLP-1 27-37 ( 0.5 g Chemmatrix Wang resin, 0.6 mmol/g)was assembled by Fmoc solid phase chemistry as for the other analogues. Following Fmoc deprotection of GIu 27, N- ⁇ -1-(4,4-Dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl-N- ⁇ - Fmoc-L-Lys [Dde-Lys(Fmoc)-OH] was coupled 3 times (1.2 mmol each) with HCTU to give a negative ninhydrin.
  • the ⁇ -N-Fmoc group was removed with 20% piperidine/DMF, and Fmoc-4-Abu (aminobutyric acid) was double-coupled with HCTU to give a negative ninhydrin.
  • Boc-Cys(Trityl) (1.5 mmol) was coupled overnight using DIC and HOBT and the ninhydrin test was negative.
  • the N- ⁇ -Dde group was removed by treating the peptidyl resin twice with 10 ml of 2% hydrazine in DMF (1 x 5 min, 1 x 8 min) and washing with DMF (1 x 10 ml), 10 % DIPEA/DMF (1 x 10 ml), and DMF (3 x 10 ml).
  • Fmoc -Ala 25 was then coupled to the resin, and the synthesis was continued through GIu 9.
  • the resin was split into 2 portions ( ⁇ .15 mmol each) and the first portion was coupled successively with Ala 8 and His 7 to give the native GLP-1 sequence with 4-Abu-Cys at the ⁇ amino of Lys 26.
  • Final Fmoc deprotection was followed by cleavage with 10 ml TFA containing H 2 O (5%), TIS (4%), thioanisole (2.5%), and EDT(0.5%) for 1.5 hr.
  • the solution was filtered into cold ether, the precipitate washed twice with cold ether, and dried under nitrogen to give 110 mg of crude SP012.
  • the product was purified by RP-HPLC and gave a mass of 3543.2, the expected MW.
  • GLP-1 glucagon-like peptide-1 ; DPP-IV, dipeptidyl-peptidase; mPEG, monomethoxy polyethylene glycol; CDI, carbonyldiimidazole; DMF, dimethylformamide; DIPEA, diisopropylethylamine; HMPA, 2-(4-hydroxymethyl phenoxy)acetic acid; DCM, dichloromethane; THF, tetrahydrofuran; Trityl, triphenylmethyl; MSNT, 1-(mesitylene- 2-sulphonyl)-3-nitro-7H-1,2,4-triazole; HCTU, 0-(1 H-6-Chlorobenzotriazole-1-yl)- 1 ,1 ,3,3-tetramethyluroniumhexaflourphosphate; TFA, trifluoroacetic acid; TIS, triisopropylsilane.

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Abstract

L'invention porte sur de nouveaux composés et sur de nouvelles compositions comprenant de tels analogues. Les composés de l'invention sont utiles dans le traitement du diabète sucré et des troubles apparentés.
PCT/GB2008/001489 2007-05-01 2008-04-29 Composés et leurs utilisations Ceased WO2008132477A1 (fr)

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GB0708445A GB2448895A (en) 2007-05-01 2007-05-01 GLP-1 like compounds and uses thereof

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103333940A (zh) * 2013-07-10 2013-10-02 浙江大学宁波理工学院 利用带鱼制备二肽基肽酶iv抑制肽的方法
CN111378028A (zh) * 2018-12-30 2020-07-07 万新医药科技(苏州)有限公司 酰化glp-1化合物及其修饰基团的合成

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005058958A2 (fr) * 2003-12-18 2005-06-30 Novo Nordisk A/S Nouveaux analogues glp-1 lies a des agents de type albumine
WO2006044063A2 (fr) * 2004-10-19 2006-04-27 Nitto Denko Corporation Administration transepitheliale de peptides avec activites des hormones incretines
WO2007022123A2 (fr) * 2005-08-11 2007-02-22 Amylin Pharmaceuticals, Inc. Polypeptides hybrides presentant des proprietes selectionnables
WO2007049940A1 (fr) * 2005-10-27 2007-05-03 Peptron Co., Ltd Conjugue a base de proteine sanguine et de substance bioactive
WO2007124461A2 (fr) * 2006-04-20 2007-11-01 Amgen Inc. Composés de glp-1

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5705483A (en) * 1993-12-09 1998-01-06 Eli Lilly And Company Glucagon-like insulinotropic peptides, compositions and methods
FR2777283B1 (fr) * 1998-04-10 2000-11-24 Adir Nouveaux composes peptidiques analogues du glucagon-peptide- 1 (7-37), leur procede de preparation et les compositions pharmaceutiques qui les contiennent
CA2412004C (fr) * 2000-06-16 2010-12-21 Eli Lilly And Company Analogues de glp-1 (glucagone-like peptide 1)
AU2003273300A1 (en) * 2002-09-06 2004-03-29 Bayer Pharmaceuticals Corporation Modified glp-1 receptor agonists and their pharmacological methods of use
CN101942018A (zh) * 2003-02-19 2011-01-12 益普生制药股份有限公司 Glp-1的类似物
PT1881850E (pt) * 2005-05-13 2010-11-26 Lilly Co Eli Compostos peguilados de glp-1

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005058958A2 (fr) * 2003-12-18 2005-06-30 Novo Nordisk A/S Nouveaux analogues glp-1 lies a des agents de type albumine
WO2006044063A2 (fr) * 2004-10-19 2006-04-27 Nitto Denko Corporation Administration transepitheliale de peptides avec activites des hormones incretines
WO2007022123A2 (fr) * 2005-08-11 2007-02-22 Amylin Pharmaceuticals, Inc. Polypeptides hybrides presentant des proprietes selectionnables
WO2007049940A1 (fr) * 2005-10-27 2007-05-03 Peptron Co., Ltd Conjugue a base de proteine sanguine et de substance bioactive
WO2007124461A2 (fr) * 2006-04-20 2007-11-01 Amgen Inc. Composés de glp-1

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
KANDA P ET AL.: "Synthesis of GLP-1 analogues as potential agents for blood glucose control", JOURNAL OF PEPTIDE SCIENCE 2006, vol. 12, no. S, 2006, pages 214, XP008094879, ISSN: 1075-2617 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103333940A (zh) * 2013-07-10 2013-10-02 浙江大学宁波理工学院 利用带鱼制备二肽基肽酶iv抑制肽的方法
CN111378028A (zh) * 2018-12-30 2020-07-07 万新医药科技(苏州)有限公司 酰化glp-1化合物及其修饰基团的合成

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