JP2008540402A - Novel compounds as GLP-1 agonists - Google Patents

Abstract

The present invention describes a group of novel peptidomimetics useful for the treatment of diabetes. These compounds are defined by the general formula (I) as follows:
AX ₁ -S ₁ -YS ₂ -X ₂ -B (I)

Description

The present invention relates to novel compounds of general formula (I), their tautomeric forms, novel intermediates involved in their synthesis, their pharmaceutically acceptable salts and pharmaceutical compositions containing them. .
AX ₁ -S ₁ -YS ₂ -X ₂ -B (I)

  In particular, the present invention relates to a novel Glucagon-Like Peptide-1 (GLP-1) peptidomimetic, which acts as a GLP-1 receptor agonist, Shows most biological activity of GLP-1. In addition, these GLP-1 peptidomimetics show high stability to proteolytic cleavage, particularly for the DPP-IV (dipeptidyl peptidase IV) enzyme, and treat diabetes and related pathologies. Alternatively, for prevention, delivery can be by invasive and various non-invasive routes of administration, including oral, nasal, buccal, pulmonary and transdermal routes of administration.

  The present invention also provides compounds of general formula (I), their tautomeric forms, their pharmaceutically acceptable salts, pharmaceutical compositions containing them, and novel intermediates involved in their synthesis It relates to the preparation method.

  GLP-1 (7-36) amide is the product of the preproglucagon gene, which is secreted from intestinal L cells in response to food intake. The physiological effects of GLP-1 are of considerable interest. GLP-1 exerts multiple actions by stimulating insulin secretion from β cells of the pancreas in a glucose-dependent manner (insulin secretion promoting action). GLP-1 also decreases circulating plasma glucagon concentration by inhibiting the secretion of plasma glucagon from α cells (Drucker D. J., Endocrinology, 142, 521-527, 2001). Recently, GLP-1 has also been shown to exhibit properties such as stimulation of β-cell proliferation, suppression of appetite, delayed gastric emptying and stimulation of insulin sensitivity (Nauck, Horm. Metab. Res., 47, 1253-1258 , 1997).

  The venom of the American lizard (Heloderma Suspectum) contains a 39 amino acid peptide called Exendin-4 (EX-4) that shares about 50% sequence identity with GLP-1 itself and is very powerful GLP-1R (glucagon-like peptide-1 receptor) agonist activity (Thorens B., Diabetes, 42, 1678-1682, 1993). In fact, EX-4 was found to be much more powerful than natural GLP-1. This is because EX-4 (25 minutes by intravenous route) has a relatively long half-life compared to GLP-1 (2-5 minutes by intravenous route). Exendin 4 binds to GLP-1R with high affinity due to the nine C-terminal sequences (Doyle M.E., Regulatory Peptides, 114, 153-158, 2003). Thus, the above pharmacological properties of GLP-1R agonists make GLP-1R agonists a highly desirable therapeutic agent for the treatment of diabetes.

  Natural or synthetic GLP-1 peptides are rapidly metabolized to inactive metabolites by proteolytic enzymes such as dipeptidyl peptidase IV (DPP-IV), thus this limits the use of GLP-1 as a drug. ing. Currently, liraglutide / NN2211 (Novonordisk; Phase III study; WO1998008871), BIM51077 (Ibsen; Phase II study; WO2000034331), CJC-1131 (conjugation; Phase II study; WO2000069911), ZP-10 Various analogs of GLP-1 and EX-4 such as (Zeeland and Aventis; Phase II trial; WO2001004156) are in different clinical development stages (Nauck MA, Regulatory Peptides, 115, 13-19, 2004) . However, all these peptides require delivery through a parenteral route of administration, such as the recently marketed BYETTA® (Exendin 4, AC2933; WO2001051078) (Amylin and Lilly) . Thus, there is a significant need to develop bioactive GLP-1 mimetics with a broad pharmacodynamic profile.

GLP-1R is a 7-transmembrane domain G protein-coupled receptor (GPCR) and is present on the plasma membrane of pancreatic β cells. The effector system of GLP-1R is the adenylyl cyclase (AC) enzyme. The interaction between GLP-1 agonist and GLP-1R causes activation of AC, which converts ATP to cAMP. Increasing intracellular cAMP levels increases the ADP / ATP ratio, thus causing cellular depolarization (due to _KATP channel closure). In addition, the activation of protein kinases (PK-A and PK-C) by increasing intracellular cAMP levels activates and activates the L-type Ca ²⁺ channel, which increases the Ca ²⁺ concentration in the systolic phase. . Increased intracellular Ca ²⁺ leads to exocytosis of insulin in pancreatic β cells (Fehmann, HC, Endocr. Rev., 16, 390-410, 1995).

  The general mechanism of interaction between peptide ligands and class B GPCRs, called the “two-domain” model, has recently been elucidated (Hoare SRJ, Drug Discovery Today, Vol. 10 (6), 417-427, 2005). In these two domain models, the C-terminal part of the peptide binds to the N domain of this receptor and the N-terminal ligand region binds to the J domain (transmembrane) region of GPCRs. This interaction activates the receptor and thereby stimulates intracellular signaling. Receptor binding and activation occur in two separate domains of exendin, but in GLP-1 they are closely linked (Eng J., JBC, 272 (34), 21291-21296, 1997).

Prior Art Previously, Bristol-Myers Squibb (BMS) (Princeton, NJ, USA) has the general formula Xaa1-Xaa11 (where Xaa1-Xaa9 is a GLP-1 peptide having some analogs. Xaa2 represents Ala or optionally substituted with Aib and Xaa6 represents Phe or optionally α-Me-Phe (2-F) -OH Reported for human GLP-1 mimetics with substituted, Xaa10 and Xaa11 represent a combination of substituted or unsubstituted biphenylalanine (Bip) derivatives (WO03 / 033671A2, US2004 / 0127423A1, WO2004 / 094461A2, US2006 / 0004222A1 and WO2006 / 014287A1).

The present invention is a novel GLP-1 peptide mimetic (hereinafter peptidomimetics) of formula (I) that acts as a GLP-1R agonist and exhibits most biological activity of the native GLP-1 peptide. I will provide a. Furthermore, these GLP-1 peptidomimetics show high stability to proteolytic cleavage, especially for the DPP-IV enzyme, and surprisingly, type 1 and type 2 diabetes, metabolic disorders, obesity and It has been discovered that it has a long half-life suitable for the treatment, alleviation and prevention of related diseases.
WO1998008871 WO2000034331 WO2000069911 WO2001004156 WO2001051078 WO03 / 033671A2 US2004 / 0127423A1 WO2004 / 094461A2 US2006 / 0004222A1 WO2006 / 014287A1 Drucker DJ, Endocrinology, 142, 521-527, 2001 Nauck, Horm. Metab. Res., 47, 1253-1258, 1997 Thorens B., Diabetes, 42, 1678-1682, 1993 Doyle ME, Regulatory Peptides, 114, 153-158, 2003 Nauck MA, Regulatory Peptides, 115, 13-19, 2004 Fehmann, HC, Endocr. Rev., 16, 390-410, 1995 Hoare SRJ, Drug Discovery Today, Vol. 10 (6), 417-427, 2005 Eng J., JBC, 272 (34), 21291-21296, 1997 G. Barany and RB Merrifield “The peptides: Analysis, synthesis, Biology” Volume 2- “Special methods in peptide synthesis, Part A” pp. 3-284, E. Gross and J. Meienhofer, Academic Press, New York, 1980 JM Stewart and JD Young `` Solid-phase peptide synthesis '' 2nd Ed., Pierce chemical Co., Rockford, II, 1984 E. Atherton and RC Sheppard “The Fluorenylmethoxycarbonyl amino protecting group”, in “The peptides: Analysis, synthesis, Biology” Volume 9- "Special methods in peptide synthesis, Part C" pp. 1-38, S. Undenfriend and J. Meienhofer, Academic Press, San Diego, 1987 Tetrahedron Letter 58, 9633-9695, 2002 Org. Letters 3 (8), 1121-1124, 2001 JOC, 2005, 70, 6918-6920 DS King et al., Int. J. peptide Protein res. 36, 1990, 255-266 Johannsson J., Clin. Endocrinol. Metab., 82, 727-34, 1997

  The present invention describes a group of novel peptidomimetics useful for the treatment of diabetes. These compounds are defined by the general formula (I) as follows: The compounds of the present invention are useful for the treatment of the human or animal body by modulating insulin secretion. Accordingly, the compounds of the present invention are suitable for the treatment, alleviation, regulation or prevention of type 1 and type 2 diabetes and obesity.

  The main object of the present invention is to provide novel compounds of the general formula (I), their tautomeric forms, novel intermediates involved in their synthesis, their pharmaceutically acceptable salts, their pharmaceuticals And pharmaceutically acceptable solvates, and pharmaceutical compositions comprising these or mixtures thereof suitable for the treatment, alleviation, or regulation of diabetes.

  In one embodiment, the preparation of the novel compounds of general formula (I), their tautomeric forms, their pharmaceutically acceptable salts, pharmaceutically acceptable solvates and pharmaceutical compositions comprising them Provide a way.

  In other embodiments, the compounds of general formula (I), their tautomeric forms, their pharmaceutically acceptable salts, solvates, and pharmaceutically acceptable carriers, solvents, diluents, Pharmaceutical compositions are provided comprising a mixture of these with forms and other media normally used in their manufacture. In yet another embodiment, as an antidiabetic agent by administering to a mammal in need of such treatment a therapeutically effective and non-toxic amount of a compound of formula (I) or a pharmaceutically acceptable composition thereof. The use of the novel compounds of the present invention is provided.

Abbreviations used The following abbreviations are used in the examples and elsewhere in this specification.
Aib = α-aminoisobutyric acid,
ACN or MeCN = acetonitrile,
Bip = biphenylalanine residue,
Bip (4-fluro) = 4-fluoro-biphenylalanine residue,
Bip (2-Me) = 2-methylbiphenyl residue,
Bip (2-Et) = 2-ethylbiphenyl residue,
Bip (2-CN) = 2-nitrile biphenyl residue,
Bip (2-Ipr) = 2-isopropylbiphenyl residue,
Bip (2'-Et-4'-OMe) = 2-ethyl-4-methoxy-biphenyl residue,
Bip (2-F) = 2-fluro-biphenyl residue,
Bn = benzyl,
Boc = tert-butoxycarbonyl,
But = O-tert-butyl group,
cAMP = adenosine 3 ′, 5 ′,-cyclic monophosphate,
DCM = dichloromethane,
DMF = N, N-dimethylformamide,
DIPCDI = di-isopropylcarbodiimide,
DIPEA = diisopropylethylamine,
4-DBF = 4-dibenzofuran-Phe-OH residue,
4-DBT = 4-dibenzothiophene-Phe-OH residue,
Dihydro-Phen = 2- (9,10-dihydro-phenanthrenyl) -Ala-OH residue,
Et = ethyl,
Et ₂ O = diethyl ether,
Fmoc = fluorenylmethoxycarbonyl,
2-Flu = 2-fluorenyl-Ala-OH residue,
g = gram,
GTT = glucose tolerance test,
GLP-1R = glucagon-like peptide-1 receptor,
h = time,
HOBt = hydroxybenzotriazole,
HOAT = 7-Aza-hydroxybenzotriazole,
HBTU = 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethylaminium hexafluorophosphate,
HPLC = high performance liquid chromatography,
L = liter,
LC / MS = liquid chromatography / mass spectrometry,
4- (2'-Me-Ph) -3-Pyr-Ala = 4- (2'-methylphenyl) -3-pyridylalanine residue,
Me = methyl,
Min = minute,
ml = ml,
μl = microliter,
mg = milligram,
mmol = mmol,
fmol = Femtomole,
MS = mass spectrometry,
1-Nap = 4- (1-naphthyl) -Phe residue,
2-Nap = 4- (2-naphthyl) -Phe residue,
Phen = 2- (phenanthrenyl) -Ala-OH residue,
Pbf = pentamethylbenzofuran-5-sulfonyl,
PyBOP = benzotriazol-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate,
SPPS = solid phase peptide synthesis,
Sc = subcutaneous,
TrPh = 4-phenyl-biphenylalanine residue,
TMS = trimethylsilyl,
TIPS = Triisopropylsilane
TFA = trifluoroacetic acid,
TBTU = 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethylaminium tetrafluoroborate,
Trt = trityl group,
(α-Me) Phe (2-F) = α-methyl-2-fluorophenylalanine residue,
-(N (Me))-= N-methylated amide bond,
D-alanine and D-Bip represented by “a” represent “D” -biphenylalanine residues;
ip = intraperitoneal,
Sequence of GLP-1 peptide = NH ₂ -HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR-CONH ₂ (30 amino acids). Thirty amino acids of the GLP-1 peptide are represented by SEQ ID NO: 1.
HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR-SEQ ID NO: 1,
Sequence of exendin = NH ₂ -HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-CONH ₂ (39 amino acids). The 39 amino acids of exendin 4 are shown in SEQ ID NO: 2.
HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-SEQ ID NO: 2.

According to the present invention, a synthetic GLP-1 analog / peptidomimetic having the structural formula (I) is provided.
AX ₁ -S ₁ -YS ₂ -X ₂ -B (I)
[In the formula, A represents an -NH-R _1, wherein R ₁ is hydrogen, a methyl group, an ethyl group, a propyl group, an isopropyl group, n- butyl group, iso - butyl, t- butyl group, pentyl group, isopentyl group, hexyl group, heptyl group, octyl group, a linear or branched (C ₁ -C ₁₅₎ groups selected from alkyl chains such as decyl, one, two or three naturally occurring amino acids amino acid or peptide comprising residues, (2-hydroxy - phenyl) - R ₃ -CO- group such as acetyl group, R _3, such as Fmoc group OC (O) - group, wherein R ₃ -SO ₂ - sulfonyl group Each of these groups may be substituted (wherein R ₃ is a methyl group, ethyl group, propyl group, isopropyl group, n-butyl group, iso-butyl group, t-butyl group, pentyl group) group, isopentyl group, hexyl group, heptyl group, octyl group, a linear or branched decyl group (C ₁ ~C ₁₀₎ alkyl , Cyclopropyl group, cyclobutyl group, cyclopentyl group, (C ₃ ~C ₆₎ cycloalkyl group such as cyclohexyl group, a phenyl group, a naphthyl group, indanyl group, a fluorenyl group, an aryl group selected from biphenyl group, a pyridyl group Selected from a heteroaryl group selected from a thienyl group, a furyl group, an imidazolyl group, a benzofuranyl group, an arylalkyl group selected from a benzyl group, a naphthylmethyl group, etc., and each of these groups may be substituted );
B represents -COOR ₂ , -CONHR ₂ or CH ₂ OR ₂ , R ₂ represents H, methyl group, ethyl group, propyl group, isopropyl group, n-butyl group, iso-butyl group, t-butyl group , pentyl group, isopentyl group, hexyl group, heptyl group, octyl group, a linear or branched (C ₁ ~C ₁₀₎ alkyl groups such as decyl, phenyl group, naphthyl group, indanyl group, a fluorenyl group, a biphenyl group Represents an aryl group selected from the above, a group selected from aralkyl groups, each of these groups may be substituted,
Each S ₁ and S ₂ independently represents a “—NH— (CH ₂ ) _n COO—” group that is a bond or n = 1-9; for example, aminoacetic acid, aminopropionic acid Derivatives of aminobutanoic acid, aminopentanoic acid, aminohexanoic acid, aminoheptanoic acid, aminooctanoic acid, aminononanoic acid, aminodecanoic acid, etc .;
Y represents a bond or —CO—, — (CH ₂ ) _m — (m = 1 to 3), “O”, “S”, —CO—NH—, —CO—NR ₄ —, natural or Represents a short peptide comprising one, two or three amino acids selected from unnatural amino acids; R ₄ is H, methyl group, ethyl group, propyl group, isopropyl group, n-butyl group, iso-butyl group, t- butyl group, a pentyl group, an isopentyl group, a hexyl group, a heptyl group, an octyl group, a linear or branched (C ₁ ~C ₁₀₎ alkyl groups such as decyl, phenyl group, naphthyl group, indanyl group Represents an optionally substituted group selected from an aryl group selected from a fluorenyl group, a biphenyl group and the like;
i) On the condition that S ₁ -YS ₂ represents a bond,
X ₁ is selected from the following amino acid sequences:
HAEGTFTSD (SEQ ID NO: 3), HAEGTFTSDV (SEQ ID NO: 4), HAEGTFTSDVS (SEQ ID NO: 5), HAEGTFTSDVSS (SEQ ID NO: 6), HAEGTFTSDVSSY (SEQ ID NO: 7), HAEGTFTSDVSSYL (SEQ ID NO: 8), HAEGTFTSDVSSYLE (SEQ ID NO: 9), HAEGTFTSDVSSYLE (SEQ ID NO: 10), HAEGTFTSDVSSYLEGQ (SEQ ID NO: 11), HAEGTFTSDVSSYLEGQA (SEQ ID NO: 12), HAEGTFTSDVSSYLEGQAA (SEQ ID NO: 13), HAEGTFTSDVSSYLEGQAAK (SEQ ID NO: 14), HAEGTFTSDVSSYLEGQAAKE (SEQ ID NO: 15), HAEGGFD SEQ ID NO: 17), which have the further option that one or more of these amino acids may be replaced by unnatural amino acids, X ₂ is the amino acid sequence
GPSSGAPPPS (SEQ ID NO: 18) or
KELEKLL (SEQ ID NO: 19) or
GPPS (SEQ ID NO: 20) or
Selected from VKGR (SEQ ID NO: 21);
ii) and S ₁ -YS ₂ does not represent a bond,
X ₁ is the following amino acid sequence, HA (SEQ ID NO: 22), HAE (SEQ ID NO: 23), HAEG (SEQ ID NO: 24), HAEGT (SEQ ID NO: 25), HAEGTF (SEQ ID NO: 26), HAEGTFT (SEQ ID NO: 27) HAEGTFTS (SEQ ID NO: 28), HAEGTFTSD (SEQ ID NO: 29), with the further option that one or more of these amino acids may be replaced by unnatural amino acids;
X ₂ is
GPSSGAPPPS (SEQ ID NO: 18) or
KELEKLL (SEQ ID NO: 19) or
GPPS (SEQ ID NO: 20) or
VKGR (SEQ ID NO: 21) or a dipeptide selected from a combination of two amino acids consisting of natural or non-natural amino acids having a side chain containing an arylalkyl or heteroarylalkyl moiety, and these arylalkyl or heteroarylalkyl moieties Benzyl group, naphthylmethyl group, pyridylmethyl group, thienylmethyl group, furylmethyl group, imidazolylmethyl group, isoxazolylmethyl group, quinolylmethyl group, benzofuranylmethyl group, benzothienylmethyl group, indolinylmethyl group , Indolylmethyl group, dibenzofuranylmethyl group, dibenzothienylmethyl group, benzodihydrofuranylmethyl group, benzodihydrothienylmethyl group, thienopyrimidylmethyl group, benzimidazolylmethyl group, phenanthrenylmethyl Selected from a group, a dihydrophenanthrenylmethyl group, a fluorenylmethyl group, a dibenzothiophenylmethyl group, etc., and in some cases, each of these groups is a methyl group, an ethyl group, a propyl group, an isopropyl group, n-butyl group, iso - butyl, t- butyl group, a pentyl group, isopentyl group, (C ₁ ~C ₆₎ alkyl groups such as hexyl group, a methoxy group, an ethoxy group, a propoxy group, a butoxy group, pentoxy group, hexanoxy group, etc. of (C ₁ ~C ₆₎ alkoxy groups, cyano, chloro group, bromo group, iodo group, a halo group such as fluoro group, hydroxy, a phenyl group, a naphthyl group, a pyridyl group, a thienyl group, a furyl group, an imidazolyl group, isoxazolyl Group, quinolyl group, benzofuranyl group, benzothienyl group, indolinyl group, indolyl group, dibenzofuranyl group, diben Depending on the case selected from thienyl group, benzodihydrofuranyl group, benzodihydrothienyl group, thienopyrimidyl group, benzimidazolyl group, phenanthrenyl group, dihydrophenanthrenyl group, fluorenyl group, dibenzofuranyl group, dibenzothiophenyl group, etc. May be substituted by an optionally substituted aryl or heteroaryl group, provided that such aryl or heteroaryl substituents are (C ₁ -C ₆ ) alkyl groups, (C ₁ -C ₆ ) With the condition that it may be further substituted by an alkoxy, cyano, halo, hydroxy or aryl or heteroaryl group].

  In a preferred embodiment, the dipeptide sequence is Bip, Bip (2-Me), Bip (2-Et), Bip (2-Ipr), Bip (2-CN), Bip (2'-Et-4'-OMe ), Bip (4′-fluoro), Bip (4′-phenyl), 2- (9,10-dihydro-phenanthrenyl) -Ala, 2- (phenanthrenyl) -Ala, 4- (2-naphthyl) -Phe, One selected from 4- (1-naphthyl) -Phe, 2-fluorenyl-Ala, 4-dibenzofuran-Phe, 4-dibenzothiophene-Phe, 4- (2′-methylphenyl) -3-pyridylalanine or May consist of multiple amino acids. The term “natural amino acid” refers to all 20 amino acids that occur in nature.

The term “unnatural amino acids” or “non-natural amino acids” refers to the substitution of D-Ala for L-Ala or D-Pro for L-Pro. Represents either the substitution of an L amino acid with an amino acid, or an appropriate modification of an L or D amino acid, aminoalkyl acid by any of the following
α-alkylation such as substitution of Ala with α-methyl Ala (Aib), substitution of Phe with α-methyl Phe, substitution of substituted Bip with α-methyl Bip;
(C ₁ ~C ₆₎ N alkylation with alkyl or (C ₃ ~C ₆₎ a group selected from cycloalkyl;
Side chain modifications such as substitution of His with histidine analogues such as 1-imidazolyl-alanine (II) or des-amino-His,

Or substitution of the phenyl ring of Phe with a pyridyl group, naphthyl group, biphenyl group;
Substitution of amino acid side chains such as substitution of aromatic amino acid side chains by halogen groups, (C ₁ -C ₃ ) alkyl groups, aryl groups, more specifically substitution of Phe by 2 and 4-halo Phe.

  Such “unnatural amino acids” or “non-natural amino acids” can generally be represented by the following structure:

Wherein R ₅ is selected from H, F, (C ₁ -C ₅ ) alkyl, and the stereochemical configuration of the carbon supporting R ₅ may be (R) or (S); R ₆ is selected from H or (C ₁ -C ₃ ) alkyl; each R ₇ and R ₈ is independently a (C ₁ -C ₃ ) alkyl or halogen atom such as H, methyl and ethyl, preferably a fluorine R ₉ is selected from atoms; R ₉ is phenyl, naphthyl, pyridyl, thienyl, furyl, imidazolyl, isoxazolyl, quinolyl, benzofuranyl, benzothienyl, indolinyl, indolyl, dibenzofuranyl , Dibenzothienyl group, benzodihydrofuranyl group, benzodihydrothienyl group, thienopyrimidyl group, benzimidazolyl group, phenanthrenyl group, dihydrophenanthrenyl group, fluorenyl group, dibenzofuranyl group, Is selected from benzothiophenyl group, (C ₁ ~C ₅₎ represents an alkyl, a group selected from aryl or Heteroriru moiety, each of these groups, optionally (C ₁ ~C ₆₎ alkyl group, (C _{1 to} C ₆ ) may be substituted by alkoxy, cyano, halo, hydroxy, optionally substituted aryl or heteroaryl, provided that such aryl or heteroaryl substituents, optionally (C ₁ ~C ₆₎ alkyl group, (C ₁ ~C ₆₎ alkoxy group, a cyano group, a halo group, that may be further substituted by hydroxy or an aryl or heteroaryl group With conditions).

  List of Fmoc protected Bip analogs used for GLP-1 peptidomimetic synthesis

  Suitable substituents include, but are not limited to, the following radicals alone or in combination with other groups. Hydroxyl, oxo, halo, thio, nitro, amino, cyano, alkyl, haloalkyl, alkoxy, haloalkoxy, cycloalkyl, aryl, aryloxy, aralkyl, aralkoxy, heteroaryl, heteroaralkyl, heteroaryloxy, heteroaralkoxy, Acyl, acyloxy, carboxylic acid, and derivatives thereof such as esters and amides.

  Various groups, groups and substituents used anywhere in this specification are described in the following paragraphs.

  The term “alkyl” as used herein, either alone or in combination with other groups, is methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl. , Amyl, t-amyl, n-pentyl, n-hexyl, iso-hexyl, heptyl, octyl, decyl, and the like, means a linear or branched group containing 1 to 10 carbons.

  The term “cycloalkyl” as used herein includes 3 to 7 carbons, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, either alone or in combination with other groups. Means group.

  The terms “aryl” or “aromatic” as used herein, either alone or in combination with other groups, are one, two, such as phenyl, naphthyl, tetrahydronaphthyl, indane, biphenyl, etc. Or it means an aromatic system comprising three rings, such rings that may be pendant-like attached or fused together.

  The term “arylalkyl” means an alkyl group attached to an aryl, such as benzyl, phenylethyl, naphthylmethyl, as defined above. The term “aryloxy” means an aryl group attached to an alkoxy group, such as phenoxy, naphthyloxy, and the like, as defined above, which may be substituted.

  The term “aralkoxy” means an allylalkyl moiety, such as benzyloxy, phenethyloxy, naphthylmethyloxy, phenylpropyloxy, as defined above, which may be substituted.

  As used herein, the term “heteroaryl” or “heteroaromatic” refers to pyridyl, thienyl, furyl, pyrrolyl, oxazolyl, triazolyl, isothiazolyl, either alone or in combination with other groups. , Imidazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzothienyl, indolinyl, indolyl, azaindolyl, azaindolinyl, benzodihydrofuranyl, benzodihydrothienyl, pyrazolopyrimidinyl, pyrazolopyrimidonyl , Azaquinazolinoyl, pyridofuranyl, pyridothienyl, thienopyrimidyl, thienopyrimidinyl, quinolinyl, pyrimidinyl, pyrazolyl, quina Linyl, quinazolonyl, pyrimidinyl, pyridazinyl, triazinyl, benzoxazinyl, benzoxazinonyl, benzothiazinyl, benzothiadinonyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, benzotriazolyl, phthalazinyl, naphthylidinyl, purinyl, carbazolyl, phenothazinyl, phenoxazinyl, etc. Comprising one or more heteroatoms selected from O, N or S, including one, two or three rings, wherein such rings are linked together in a pendant manner or fused together An aromatic system that may be present.

  The term “heteroaralkyl” as used herein, as defined above, either alone or in combination with other groups, (2-furyl) methyl, (3-furyl) methyl, ( Linear or branched saturation containing 1-6 carbons, such as 2-thienyl) methyl, (3-thienyl) methyl, (2-pyridyl) methyl, 1-methyl-1- (2-primidyl) ethyl Means a heteroaryl group attached to a carbon chain. The terms “heteroaryloxy”, “heteroaralkoxy” and “heterocyclooxy” mean heteroaryl and heteroarylalkyl bonded to an oxygen atom, respectively, as defined above.

  The term “acyl” as used herein, either alone or in combination with other groups, includes 1-formyl, acetyl, propanoyl, butanoyl, iso-butanoyl, pentanoyl, hexanoyl, heptanoyl, benzoyl, and the like. Means a group containing ˜8 carbons, which may be substituted.

  The term “carboxylic acid” as used herein, alone or in combination with other groups, means a —COOH group and includes derivatives of carboxylic acids such as esters and amides. The term “ester” as used herein, alone or in combination with other groups, means a —COO— group, where the ester moiety may be substituted with alkoxy such as methoxycarbonyl, ethoxycarbonyl and the like. In the case of carbonyl, a carboxylic acid derivative is included.

  Unless otherwise stated, the term “amino acid” as used herein, alone or as part of another group, includes amino and carboxyl groups attached to the same carbon, referred to as the “α” carbon. It is not limited to.

  The absolute “S” configuration at the “α” carbon is commonly referred to as the “L” or natural configuration. The “R” configuration at the “α” carbon is commonly referred to as the “D” amino acid. If both “α-substituents” such as hydrogen or methyl are identical, the amino acid is Gly or Aib and not chiral.

  The term “receptor modulator” refers to a compound that acts at the GLP-1 receptor to alter its ability to control downstream signaling. Examples of receptor modulators are agonists, partial agonists, inverse agonists, allosteric potentiators and the like.

  Preferably, the isolated peptidomimetic is 3-30 mer, and such a peptide binds to and activates the GLP-1 receptor.

  According to the present invention, the synthetic isolated peptidomimetics described herein have the ability to mimic the biological activity of GLP-1 peptides in preference to agonist activity at GLP-1R. These synthetic peptidomimetic GLP-1 mimics exhibit the desired in-vivo properties, thus making them ideal therapeutic candidates for oral or parenteral administration.

  The present invention provides pharmaceutical compositions of compounds of formula (I) using such compounds, either alone or in combination, and provides methods of using such compounds. In particular, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) alone or in combination with a pharmaceutically acceptable carrier.

In addition, a therapeutically effective amount of a compound of formula (I) or a combination thereof is administered to a mammal (e.g., human) that is a patient in need of treatment to treat diabetes, particularly type II diabetes (retinopathy, neuropathy, nephropathy and Including diabetes complications such as delayed wound healing), as well as insulin resistance (glucose homeostasis), hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, obesity, hyperlipidemia (hypertriglyceridemia) Etc.), related syndromes such as X syndrome, atherosclerosis and hypertension, or methods of delaying progression or onset.
Several synthetic routes well known to those skilled in the art of peptide synthesis can be used to prepare the compounds of the invention. Compounds of formula (I) with all abbreviations pre-defined are synthesized using the methods described below, together with conventional techniques well known to those skilled in the art of peptide synthesis or various applications recognized by those skilled in the art be able to. Related methods include, but are not limited to, the following methods.

  The peptidomimetics described herein are described in G. Barany and RB Merrifield “The peptides: Analysis, synthesis, Biology” Volume 2- “Special methods in peptide synthesis, Part A” pp. 3-284, E. Gross And J. Meienhofer, Academic Press, New York, 1980; and JM Stewart and JD Young `` Solid-phase peptide synthesis '' 2nd Ed., Pierce chemical Co., Rockford, II, 1984. It can be produced by chemical synthesis using suitable applications of various solid phase methods.

A preferred strategy for preparing the peptidomimetics of the present invention is that of the Fmoc-based SPPS approach, where the Fmoc (9-fluorenyl-methyl-methyloxycarbonyl) group is used for temporary protection of the α-amino group. Based on use, this method is an acid labile protecting group for the temporary protection of amino acid side chains, such as t-butyloxycarbonyl (Boc), tert-butyl (Bu ^t ) Used in combination with a trityl (Trt) group (see e.g., E. Atherton and RC Sheppard "The Fluorenylmethoxycarbonyl amino protecting group", in "The peptides: Analysis, synthesis, Biology" Volume 9- " Special methods in peptide synthesis, Part C "pp. 1-38, edited by S. Undenfriend and J. Meienhofer, Academic Press, San Diego, 1987).

  Examples of orthogonally protected amino acids used in Fmoc solid phase peptide synthesis for the synthesis of peptidomimetics

  Peptidomimetics can be synthesized in a step-wise manner against an insoluble polymer support (resin) starting from the C-terminus of the peptide. In one embodiment, synthesis is initiated by adding the C-terminal amino acid of the peptide to the resin through formation of an amide, ester or ether bond. This can finally release the peptides obtained as C-terminal amides, carboxylic acids or alcohols, respectively.

  In Fmoc-based SPPS, the C-terminal amino acid used in the synthesis and all other amino acids are protected in their alpha amino groups and in different forms (orthogonal protection) side chain functionality (existence) These are, for example, 20% without premature cleavage of the peptide from the resin or deprotection of side-chain protecting groups usually protected with acid labile protecting groups. The α-amino protecting group can be selectively removed during synthesis using a suitable base such as a piperidine solution.

  Amino acid coupling is performed by activation of the carboxyl group as an active ester and its reaction with an unblocked alpha amino group of the N-terminal amino acid added to the resin. After all coupling and deprotection, the peptidyl-resin was washed with a large amount of solvent such as DMF, DCM and diethyl ether. The series of α-amino group deprotection and coupling is repeated until the desired peptide sequence is assembled. The peptide is then removed from the resin using an appropriate cleavage mixture, in parallel with side-chain functional deprotection, usually in the presence of an appropriate scavenger to limit side reactions. Cleave. The resulting peptide is finally purified by reverse phase HPLC.

  The synthesis of the peptidyl-resins required as final peptide precursors utilizes commercially available cross-linked polystyrene polymer resins (Novabiochem, San Diego, CA). Desirable for use in the present invention is Fmoc-PAL-PEG-PS resin, 4- (2 ′, 4′-dimethoxyphenyl-Fmoc-aminomethyl) -phenoxyacetyl-p-methylbenzhydrylamine resin (Fmoc- Rink amide MBHA resin), 2-chloro-trityl-chloride resin or p-benzyloxybenzyl alcohol resin (HMP resin), which may or may not have a C-terminal amino acid already attached to them. . If the C-terminal amino acid is not bound, it can be bound by the HOBt active ester of the Fmoc protected amino acid formed by reaction with DIPCDI. In the case of 2-chloro-trityl resin, the first Fmoc protected amino acid coupling was achieved using DIPEA. In the next amino acid assembly, N-terminal protection of the peptidyl resin was selectively deprotected using a 10-20% solution of piperidine solution. After all coupling and deprotection, excess amino acids and coupling agents were removed by washing with DMF, DCM and ether. Subsequent amino acid coupling can be achieved using HOBt or HOAT (from DIPCDI / HOBt or DIPCDI / HOAT, respectively) active esters. For some difficult couplings, especially with amino acids that are hydrophobic or have large side chain protection, complete coupling can be achieved with additives such as DIPEA and HBTU, PyBOP or TBTU. This can be achieved by the combined use with a highly efficient coupling agent.

  General scheme for Fmoc-based SPPS

  The peptide analogs described herein can be synthesized using a batch or continuous flow peptide synthesizer. Non-natural, non-commercial amino acids present at different positions were attached to the peptide chain using one or more methods known in the art. In one approach, Fmoc protected natural amino acids were prepared in solution using appropriate literature procedures. For example, the Fmoc protected Bip analog described above was prepared using a modified Suzuki cross coupling method known in the literature (eg, Tetrahedron Letter 58, 9633-9695, 2002). Fmoc protected α-methylated amino acids were prepared using the asymmetric Strecker synthesis method described in Org. Letters 3 (8), 1121-1124, 2001. Fmoc protected N-methylated amino acids were prepared using literature methods described, for example, in JOC, 2005, 70, 6918-6920. The resulting derivative was then used for the stepwise synthesis of peptides. Alternatively, the required unnatural amino acids were constructed directly on the resin using synthetic organic chemistry procedures to construct linear peptides.

The peptide-resin precursor of each peptidomimetic can be obtained from any of the standard cleavage procedures described in the literature (e.g., DS King et al., Int. J. peptide Protein res. 36, 1990, 255-266). Cleavage and deprotection can be performed using appropriate application methods. A preferred method for use in the present invention is the use of a TFA cleavage mixture in the presence of water and TIPS as a scavenger. In general, peptidyl-resins were incubated in TFA / water / TIPS (94: 3: 3; V: V: V; 10 ml / peptidyl resin 100 mg) for 1.5-2 hours at room temperature. The cleaved resin is then filtered and the TFA solution is concentrated or dried under reduced pressure. The resulting crude peptide is washed with precipitation or Et ₂ O or redissolved directly in DMF or 50% aqueous acetic acid for purification by preparative HPLC.

Peptidomimetics with the desired purity can be obtained by purification using preparative HPLC. The crude peptide solution was injected into a semi-Prep column (Luna10μ; _C18 ; 100A °) with dimensions of 250 x 50 mm, using a flow rate of 15-50 ml / min with effluent monitoring by a PDA detector at 220 nm. Elute with a linear gradient of ACN in water (both buffered with 0.1% TFA). The structure of the purified peptidomimetic can be confirmed by electrospray mass spectrometry (ES-MS).

  After purification by preparative HPLC, all prepared peptides were isolated as trifluoroacetate salts by TFA as counterion. However, some peptides were desalted by passing through a suitable ion exchange resin bed, preferably the anion exchange resin Dowex SBR P (Cl) or equivalent basic anion exchange resin. In some examples, TAF counter ions were replaced with acetate ions by passing through a suitable ion exchange resin eluted with dilute acetic acid solution. For the preparation of peptide hydrochloride in the final stage of manufacture, selected peptides were treated with 4M HCl along with acetate. The resulting solution was filtered through a membrane filter (0.2 μm) and then lyophilized to obtain a white to off-white HCl salt. Other suitable pharmaceutically acceptable salts of the peptidomimetics of the present invention were prepared according to similar techniques and / or such appropriate modifications that are within the skill of the art.

  In a preferred embodiment, the invention provides a method of making a peptidomimetic that mimics the activity of an endogenous polypeptide GLP-1R agonist. In another preferred embodiment, the polypeptide receptor agonist is GLP-1.

  The novel compounds of the present invention can be formulated into suitable pharmaceutically acceptable compositions by combining with known appropriate excipients.

  The pharmaceutical composition is provided by using conventional techniques. Preferably, this composition is in unit dosage form containing the active ingredient, ie either a compound of formula (I) according to the invention, alone or in combination.

  The amount of active ingredient in the pharmaceutical composition and unit dosage form (i.e., the compound of formula (I) described in this invention) varies widely depending on the particular application, the potency of the particular compound and the desired concentration, or Can be adjusted. When used for the indicated effect for guidance, the daily oral dose of the active ingredient is between about 0.001 and 1,000 mg / kg / body weight, preferably about 0.01 to 100 mg / kg / body weight Most preferably in the range of about 0.6 to 20 mg / kg / day.

<General preparation of peptidomimetics using SPPS approach>
(Assembly of peptide mimetic to resin)
A sufficient amount (50-100 mg) of Fmoc-PAL-PEG-PS resin or Fmoc-Rink amide MBHA resin (throughput 0.5-0.6 mmol / g) was added 2-10 in DMF (1-10 ml / 100 mg resin). Inflated for minutes. The Fmoc group on the resin was then removed by incubation of the resin with 10-30% piperidine in DMF (10-30 ml / 100 mg resin) for 10-30 minutes. The deprotected resin was filtered and washed with copious amounts of DMF, DCM and ether (50 ml × 4). The washed resin was incubated for 5 minutes in freshly distilled DMF (1 ml / resin 100 mg) under a nitrogen atmosphere. A 0.5M solution of the first Fmoc protected amino acid (1-3 eq.) Pre-activated with HOBt (1-3 eq.) And DIPCDI (1-2 eq.) In DMF is added to the resin, and then the resin is added to a nitrogen atmosphere. Shake for 1-3 hours under. Coupling completion was monitored using a qualitative ninhydrin test. After coupling of the first amino acid, the resin was washed with DMF, DCM and diethyl ether (50 ml × 4). In the next amino acid coupling, first the Fmoc protection on the first amino acid coupled to the resin is deprotected with a 10-20% piperidine solution, followed by the use of an appropriate coupling agent, The second Fmoc protected amino acid was also coupled as described above. The deprotection, water wash, coupling and water wash cycles were repeated until the desired peptide chain was assembled on the resin as in the general scheme above.

  Finally, the Fmoc protected peptidyl-resin prepared as described above was deprotected by treatment with 20% piperidine as described above and the peptidyl-resin was washed with DMF, DCM and diethyl ether (50 ml × 4). The resin containing the desired peptide was dried for 10-15 minutes under nitrogen pressure to perform cleavage / deprotection.

(Cleavage and deprotection)
The desired peptidomimetics were cleaved and deprotected from their respective peptidyl-resins by treatment with a TFA cleavage mixture as described below. A solution of TFA / water / triisopropylsilane (95: 2.5: 2.5) (10 ml / 100 mg peptidyl-resin) was added to the peptidyl-resin and the mixture was left at room temperature with occasional stirring. The resin was filtered and washed with the cleavage mixture, and the combined filtrates were evaporated to dryness. The obtained residue was dissolved in 10 ml of water, the aqueous layer was extracted three times with ether (20 ml each), and finally the aqueous layer was lyophilized. The crude peptide obtained after lyophilization was purified by preparative HPLC as described below.

(Preparative HPLC purification of crude peptide mimetics)
Preparative HPLC was performed on a Shimadzu LC-8A liquid chromatograph. A solution of the crude peptide dissolved in DMF or water was injected into a semi-Prep column (Luna 10μ; C ₁₈ ; 100A °) with a size of 250 × 50 mm, and the effluent was monitored by a PDA detector at 220 nm, while 15-50 ml / Elute with a linear gradient of ACN in water (both buffered with 0.1% TFA) using a flow rate of minutes. A typical 20% to 70% gradient of a water-CAN mixture buffered with 0.1% TFA was used over 50 minutes with a gradient change of 1% per minute. The desired product that was eluted was collected in fractions of 10-20 ml, and pure peptidomimetics were obtained by lyophilization of each HPLC fraction as an amorphous white powder.

<HPLC analysis of purified peptidomimetics>
After purification by preparative HPLC as described above, each peptide was analyzed by analytical RP-HPLC on a Shimadzu LC-10AD analytical HPLC system. For analytical HPLC analysis of peptidomimetics, a Luna 5μ; C ₁₈ ; 100 A °; 250 x 4.6 nm column was used with a linear gradient of 0.1% TFA and ACN buffer, and chromatogram capture was performed using a PDA detector. At 220 nm.

<Characteristic determination by mass spectrometry>
Each peptide was characterized by electrospray ionization mass spectrometry (ESI-MS) in either flow injection or LC / MS mode. A triple quadrupole mass spectrometer (API-3000 MDS-SCIES, Canada) was used for all analyzes in positive and negative ion electrospray mode. Complete scan data was obtained over the quadrupole mass range, operated at unit resolution. In all cases, the experimentally determined molecular weight was within 0.5 Daltons of the calculated monoisotopic molecular weight. Mass chromatogram quantification was performed using Analyst 1.4.1 software.

  The following GLP-1 peptidomimetics were prepared using the synthetic methods described herein in conjunction with other commonly known techniques and their appropriate applications. This list represents a group of various peptidomimetics, which can be prepared according to the present invention and are expected to include at least obvious variations of these compounds. However, such disclosure is not to be construed as limiting the scope of the invention in any way.

  The following compounds can be prepared according to the general methods described above and are encompassed within the scope of the present invention (Tables 7-9).

Peptidomimetics prepared as described above were tested for in vitro GLP-1 agonist activity using the following cAMP cell line assay. GLP-1 mimetic peptide analogs stimulated cAMP production in a dose-response manner, and corresponding EC ₅₀ values were measured with some selected peptidomimetics that showed activity in the range of 10-100 nM in vitro . EC ₅₀ values of EX-4, was used as a positive control.

<Cyclic AMP measurement>
The GLP-1 receptor is a G protein coupled receptor. The biologically active form of GLP-1 (7-36) amide binds to the GLP-1 receptor, triggers adenylate cyclase activation through signaling and increases intracellular cAMP levels. In order to monitor the agonism of peptide compounds in stimulating the GLP-1 receptor, adenyl cyclase activity was monitored by quantifying cellular cAMP levels.

(CAMP assay)
For cAMP production, stably transfected CHO / HGLP1R cells were quantified on a semi-high throughput platform using the DiscoverX cAMP kit with exendin 4 as a positive control. NCE activity was measured as% exendin 4 activity at a concentration of 0.01 μM. Positive compounds were further verified for cAMP production using an indirect cAMP ELISA kit (R & D system). The activity of this compound was expressed as fmol cAMP / μg protein. EC ₅₀ values for some representative compounds (I-IV) are shown in FIG.

(Demonstration of in vivo efficacy of compounds)
The in vivo glucose lowering properties of some representative compounds in animal models are described below. This study was used to examine the in vivo efficacy of the compounds of the present invention against blood glucose at hyperglycemia. Intraperitoneal glucose tolerance test (IPGTT) was performed on Swiss albino mice (SAM) fasted overnight with a body weight of 25-30 g. Mice were given a glucose load of 1.5 g / Kg / 10 ml and blood was collected at different time intervals from the retroorbital plexus. The test compound (peptidomimetic) was dissolved in an appropriate nmol / ml concentration of solvent equivalent to the dose administered at nmol / kg so that each mouse received the same volume / dose solution weight.

  Blood samples were taken before vehicle / test compound / glucose loading (0 minutes) and then after 15, 30, 60 and 120 minutes. The vehicle / test compound was administered via the intraperitoneal route 15 minutes prior to glucose loading. The blood sample was centrifuged and the resulting serum was stored at -20 ° C for analysis. The test compound was tested with a standard (positive control) and vehicle control in 6 animals in each group. Glucose was measured from serum by the GOD / POD method. The average of the two results was calculated. The absolute value of serum glucose concentration was calculated using MS Excel software. A line graph corrected with a 0 minute baseline was plotted using Graphpad prism software (ver 3.0). Calculate area under the curve (AUC) and area under the baseline correction curve (Baseline corrected area under the curve: BCAUC), one-way analysis of variance followed by Dunnet's post-test using Graphpad prism software (ver 3.0) ( The analysis was performed by conducting Dunnett's post test.

Using the experimental protocol described above, the in vivo glucose lowering properties of several selected compounds exhibiting an EC ₅₀ in vitro were measured by the CHO-GLP-1R cAMP assay in the range of 1-50 nM. In Table 10, the in vivo glucose lowering properties (ED _{50 in the} IPGTT SAM model) of four representative compounds selected (compounds I, II, III and IV) are shown.

  Several compounds of the present invention were screened in vivo using other animal models (e.g., ob / ob, db / db and C57), which showed in vivo efficacy and varying degrees of efficacy. showed that.

<Usefulness>
The present invention provides novel GLP-1 peptidomimetics that show a tendency to mimic GLP-1, such that the compounds of the present invention have agonist activity at the GLP-1 receptor. Furthermore, many GLP peptidomimetics of the present invention exhibit increased stability against proteolytic cleavage compared to the GLP-1 native sequence.

  Thus, the compounds of the present invention can be administered to mammals, preferably humans, for the treatment of various conditions and disorders, which are diabetic (preferably type II, impaired glucose tolerance, insulin resistance, Diabetic complications such as nephropathy, retinopathy, neuropathy and cataract), hyperglycemia, hyperinsulinemia, hypercholesterolemia, elevated blood levels of free fatty acids or glycerol, hyperlipidemia, hypertriglyceridemia, obesity Treatment of wound healing, tissue ischemia, atherosclerosis, hypertension, bowel disease (such as necrotizing enterocolitis, microvillous inclusion disease or celiac disease), or delaying progression or onset, It is not limited. Moreover, the compound of the present invention can increase the blood concentration of high density lipoprotein (HDL).

  Furthermore, conditions and diseases collectively referred to as “X syndrome” or metabolic syndrome, as detailed in Johannsson J., Clin. Endocrinol. Metab., 82, 727-34, 1997, are also included in the present invention. The compound can be used to treat. In some cases, the compounds of the present invention can be used in combination with an appropriate DPP-IV inhibitor for the treatment of some of the above disease states, either by sequential administration of the compound or by appropriate It is used in combination as a preparation containing the compound of the present invention together with a DDP-IV inhibitor.

  No adverse effects were observed on any of the mentioned inventive compounds. The compounds of the present invention showed good serum glucose concentration lowering activity in the experimental animals used. These compounds are used for the examination / prevention of diseases caused by hyperinsulinemia, hyperglycemia such as NIDDM, metabolic disorders and obesity. This is because such diseases are related to each other.

Claims (14)

An isolated polypeptide having the sequence of formula (I), including its tautomers, solvates and pharmaceutically acceptable salts
AX ₁ -S ₁ -YS ₂ -X ₂ -B (I)
[Where
A represents -NH-R ₁ , where R ₁ is hydrogen, a linear or branched (C ₁ -C ₁₅ ) alkyl chain, an amino acid containing one, two or three natural amino acid residues Or a group selected from a peptide, a R ₃ —CO— group, a R ₃ OC (O) — group, a sulfonyl group of the formula R ₃ —SO ₂ —, each of which may be substituted; R ₃ is selected from a linear or branched (C ₁ -C ₁₀ ) alkyl group, (C ₃ -C ₆ ) cycloalkyl group, aryl group, heteroaryl group, arylalkyl group, and each of these groups is substituted May be;
Is B, -COOR _2, represents -CONHR ₂ or CH ₂ OR _2, R ₂ is, H, linear or branched (C ₁ ~C ₁₀₎ alkyl group, a phenyl group, a naphthyl group, indanyl group, a fluorenyl Group, an aryl group selected from a biphenyl group, a group selected from an aralkyl group, wherein the aryl group is as defined above, and each of these groups may be substituted;
Each S ₁ and S ₂ independently represents a bond or a “—NH— (CH ₂ ) _n —COO—” group _where n = 1-9;
Y represents a bond or —CO—, — (CH ₂ ) _m — (m = 1 to 3), “O”, “Sr”, —CO—NH—, —CO—NR ₄ —, natural or Represents a short peptide comprising one or two or three amino acids selected from unnatural amino acids; R ₄ is H, a linear or branched (C ₁ -C ₁₀ ) alkyl group, or a phenyl group; Represents an optionally substituted aryl group selected from a naphthyl group, an indanyl group, a fluorenyl group, a biphenyl group; provided that when S ₁ -YS ₂ represents a bond,
X ₁ is selected from the following amino acid sequences:
HAEGTFTSD, HAEGTFTSDV, HAEGTFTSDVS, HAEGTFTSDVSS, HAEGTFTSDVSSY, HAEGTFTSDVSSYL, HAEGTFTSDVSSYLE, HAEGTFTSDVSSYLEG, HAEGTFTSDVSSYLEGQ, HAEGTFTSDVSSYLEGQA, HAEGTFTSDVSSYLEGQAA, HAEGTFTSDVSSYLEGQAAK, HAEGTFTSDVSSYLEGQAAKE, HAEGTFTSDVSSYLEGQAAKEF, HAEGTFTSDVSSYLEGQAAKEFI, 1 or more of these amino acids, that may be substituted by non-natural amino acids With further options, X ₂ is selected from the following amino acid sequences:
GPSSGAPPPS or
KELEKLL or
GPPS or
VKGR;
And if S ₁ -YS ₂ does not represent a bond,
X ₁ is selected from the following amino acid sequences:
HA, HAE, HAEG, HAEGT, HAEGTF, HAEGTFT, HAEGTFTS, HAEGTFTSD with the further option that one or more of these amino acids may be replaced by unnatural amino acids;
X ₂ is selected from:
GPSSGAPPPS or
KELEKLL or
GPPS or
VKGR or a dipeptide selected from a combination of two amino acids consisting of natural or non-natural amino acids having a side chain containing an arylalkyl or heteroarylalkyl moiety; these arylalkyl or heteroarylalkyl moieties are benzyl, naphthylmethyl groups , Pyridylmethyl, thienylmethyl, furylmethyl, imidazolylmethyl, isoxazolylmethyl, quinolylmethyl, benzofuranylmethyl, benzothienylmethyl, indolinylmethyl, indolylmethyl, dibenzofura Nylmethyl group, dibenzothienylmethyl group, benzodihydrofuranylmethyl group, benzodihydrothienylmethyl group, thienopyrimidylmethyl group, benzimidazolylmethyl group, phenanthrenylmethyl group, dihydrophenanthate Nirumechiru group, fluorenylmethyl group, dibenzofuranyl methyl, is selected from dibenzothiophenyl methyl group, wherein each of these groups, (C ₁ ~C ₆₎ alkyl, (C ₁ ~C ₆₎ It may be substituted by an alkoxy group, a cyano group, a halo group, a hydroxy group, or an optionally substituted aryl or heteroaryl group provided that such aryl or heteroaryl substituents are (C _1- C ₆ ) alkyl group, (C ₁ -C ₆ ) alkoxy group, cyano group, halo group, hydroxy group, or aryl or heteroaryl group. The compound of claim 1, wherein the unnatural amino acid is represented by general formula (IIa)

Wherein R ₅ is selected from H, F, (C ₁ -C ₅ ) alkyl, and the stereochemical configuration at the carbon bearing R ₅ may be (R) or (S); R ₆ is selected from H or (C ₁ -C ₃ ) alkyl; each R ₇ and R ₈ is independently selected from H, (C ₁ -C ₂ ) alkyl or a halogen atom, preferably a fluorine atom. R ₉ is a (C ₁ -C ₅ ) alkyl moiety, phenyl group, naphthyl group, pyridyl group, thienyl group, furyl group, imidazolyl group, isoxazolyl group, quinolyl group, benzofuranyl group, benzothienyl group, indolinyl group, indolyl Group, dibenzofuranyl group, dibenzothienyl group, benzodihydrofuranyl group, benzodihydrothienyl group, thienopyrimidyl group, benzimidazolyl group, phenanthrenyl group, dihydrophenanthrenyl group, fluorenyl group, dibenzofuranyl group, Is selected from benzothiophenyl group, it represents a group selected from aryl or Heteroriru moiety, each of these groups are, (C ₁ ~C ₆₎ alkyl, (C ₁ ~C ₆₎ alkoxy group, a cyano group, It may be substituted by a halo group, a hydroxy group or an optionally substituted aryl or heteroaryl group provided that such aryl or heteroaryl substituents are (C ₁ -C ₆ ) alkyl groups. , (C ₁ -C ₆ ) with the condition that it may be further substituted by an alkoxy group, a cyano group, a halo group, a hydroxy group, or an aryl or heteroaryl group). Said dipeptide representing X ₂ is preferably Bip group, Bip (2-Me) group, Bip (2-Et) group, Bip (2-Ipr) group, Bip (2-CN) group, Bip (2 '-Et-4'-OMe) group, Bip (4'-fluoro) group, Bip (4'-phenyl) group, 2- (9,10-dihydro-phenanthrenyl) -Ala group, 2- (phenanthrenyl)- Ala group, 4- (2-naphthyl) -Phe group, 4- (1-naphthyl) -Phe group, 2-fluorenyl-Ala group, 4-dibenzofuran-Phe group, 4-dibenzothiophene-Phe group, 4- ( 2. An isolated peptide according to claim 1, selected from 2'-methylphenyl) -3-pyridylalanine groups.   The substituent is hydroxyl, oxo, halo, thio, nitro, amino, cyano, alkyl, haloalkyl, alkoxy, haloalkoxy, cycloalkyl, aryl, aryloxy, aralkyl, aralkoxy, heteroaryl, heteroaralkyl, heteroaryloxy, hetero 2. The isolated peptide of claim 1 selected from aralkoxy, acyl, acyloxy, carboxylic acid, and derivatives thereof selected from esters and amides. 2. The isolated polypeptide according to claim 1, wherein the isolated polypeptide is a compound selected from:

2. The isolated polypeptide according to claim 1, wherein the isolated polypeptide is a compound selected from:

  7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6 prepared according to the method described herein and a suitable pharmaceutically acceptable carrier.   8. A compound or pharmaceutical composition according to any one of claims 1 to 7 which has the ability to mimic the biological activity of GLP-1, more preferably mimic GLP-1R agonist activity.   8. The compound or pharmaceutical composition according to any one of claims 1 to 7, which is useful for the treatment and prevention of diseases in which GLP-1R peptide performs pathophysiological functions.   Hyperlipidemia, hypercholesterolemia, hyperglycemia, hyperinsulinemia, elevated blood levels of free fatty acids or glycerol, hypertriglyceridemia, wound healing, obesity, impaired glucose tolerance, leptin resistance, insulin resistance, diabetes A method for the prevention or treatment of diseases caused by complications (e.g. nephropathy, retinopathy, neuropathy and cataract), comprising a compound of formula (I) as defined in any one of claims 1 to 7 Administering an effective non-toxic amount to a patient in need thereof.   The underlying diseases are type 2 diabetes, impaired glucose tolerance, dyslipidemia, hypertension, obesity, atherosclerosis, hyperlipidemia, coronary artery disease, cardiovascular disease, and insulin resistance are the underlying pathophysiological mechanisms 11. The method according to claim 10, which is another disease.   Administering a compound of formula (I), a pharmaceutically acceptable carrier, diluent, excipient or solvate as defined in any one of claims 1 to 7 to a patient in need thereof. An agent for treating / ameliorating any disease state as defined in any one of claims 9 to 11 comprising:   12.A method according to any one of claims 9 to 11, comprising administering a compound of formula (I) as defined in any one of claims 1 to 7 to a patient in need thereof in combination with a suitable DPP IV inhibitor. An agent for treating / alleviating a disease state as defined in one paragraph.   Use of a compound of formula (I) alone or in combination with a suitable DPP IV inhibitor as a drug suitable for the treatment of a disease as defined in any one of claims 9 to 11, and Use of pharmaceutical compositions and medicaments comprising them according to any of claims 7 to 9, 12 and 13.