EP1891106A2 - Novel compounds as glp-i agonists - Google Patents
Novel compounds as glp-i agonistsInfo
- Publication number
- EP1891106A2 EP1891106A2 EP06809915A EP06809915A EP1891106A2 EP 1891106 A2 EP1891106 A2 EP 1891106A2 EP 06809915 A EP06809915 A EP 06809915A EP 06809915 A EP06809915 A EP 06809915A EP 1891106 A2 EP1891106 A2 EP 1891106A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- bip
- ome
- pyr
- ala
- aib
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- 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
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
- A61P27/12—Ophthalmic agents for cataracts
-
- 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
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Definitions
- 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.
- the present invention relates to novel Glucagon-Like Peptide- 1 (GLP-I) peptide mimics (peptidomimetic), which act as GLP-I receptor agonists and exhibit most of the biological activity of the native GLP-I.
- GLP-I peptidomimetics exhibit increased stability to proteolytic cleavage, especially against DPP-IV (Dipeptidyl peptidase-IV) enzyme and can be delivered by both invasive and various non-invasive routes of administrations such as oral, nasal, buccal, pulmonary and transdermal route of administration, for the treatment or prevention of diabetes and related conditions.
- the present invention also relates to a process of preparing compounds of general formula (I), their tautomeric forms, their pharmaceutically acceptable salts, pharmaceutical compositions containing them, and novel intermediates involved in their synthesis.
- the GLP-I (7-36) amide is a product of the preproglucagon gene, which is secreted from intestinal L-cells, in response to the ingestion of food.
- the physiological action of GLP-I has gained considerable interest. GLP-I exerts multiple action by stimulating insulin secretion from pancreatic ⁇ -cells, in a glucose dependent manner (insulinotropic action). GLP-I also lowers circulating plasma glucagon concentration, by inhibiting its secretion from ⁇ -cells (Drucker D. J., Endocrinology, 142, 521-527, 2001). More recently, it has become clear that GLP-I also exhibits properties like stimulation of ⁇ -cell growth, appetite suppression, delayed gastric emptying and stimulation of insulin sensitivity (Nauck, Horm. Metab. Res., 47, 1253-1258, 1997).
- the venom of the GiIa Monster Heloderma Suspectuni contains a 39 amino acid peptide called Exendin-»4 (EX-4) that shares around 50 % sequence identity to GLP-I itself, exhibits a very potent GLP-IR (Glucagon like peptide-1 receptor) agonist activity (Thorens B., Diabetes, 42, 1678 - 1682, 1993). Indeed, it was found that EX-4 is much more potent than native GLP-I peptide, because of its relatively longer half- life (25 min., iv route of administration), compared to GLP-I (2-5 min., iv route of administration).
- EX-4 Glucagon like peptide-1 receptor
- Exendin-4 binds with greater affinity to the GLP-IR, due to presence of the nine extra C-terminal sequence (Doyle M.E., Regulatory Peptides, 114, 153-158, 2003).
- Doyle M.E. Regulatory Peptides, 114, 153-158, 2003.
- Native or synthetic GLP-I peptide is rapidly metabolized by the proteolytic enzymes, such as dipeptidyl peptidase-IV (DPP-IV) into an inactive metabolite, thereby limiting the use of GLP-I as a drug.
- DPP-IV dipeptidyl peptidase-IV
- GLP-I and EX-4 are in different stages of clinical development (Nauck M. A., Regulatory Peptides, 115, 13-19, 2004).
- the GLP-I R is a seven-transmembrane domain G-protein-coupled receptor (GPCR) and it is located on the cell membrane of pancreatic ⁇ -cells.
- the effector system of GLP-I R is the Adenylyl Cyclase (AC) enzyme. Interaction of GLP-I agonist with GLP-IR causes activation of AC, which converts ATP to cAMP. Increase in the intracellular cAMP level raises the ratio of ADP/ATP, thereby initiating the cell depolarization (due to closure of K ATP channel).
- PK-A & PK-C Protein Kinase
- cystolic Ca 2+ concentration by opening of L-type of Ca 2+ channel.
- An increase in the intracellular Ca 2+ leads to exocytosis of insulin, in pancreatic ⁇ -cells (Fehmann, H.C., Endocr. Rev., 16, 390 - 410, 1995).
- peptidomimetics which act as a GLP-IR agonist and exhibit most of the biological activity of the native GLP-I peptide. Furthermore, these GLP-I peptidomimetics exhibit increased stability to proteolytic cleavage, especially against DPP-IV enzyme and therefore, surprisingly found to have an increased half-life making them suitable for the treatment / mitigation / prophylaxis of both type 1 & type 2 diabetes, metabolic disorders, obesity and related disorders. Summary of the invention
- 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 given below.
- the compounds of the present invention are useful in the treatment of the human or animal body, by regulation of insulin secretion. The compounds of this invention are therefore suitable for the treatment/mitigation/regulation or prophylaxis of both type 1 & type 2 diabetes and obesity.
- the main object of the present invention is to provide novel compounds of general formula (I), their tautomeric forms, novel intermediates involved in their synthesis, their pharmaceutically acceptable salts, their pharmaceutically acceptable solvates and pharmaceutical compositions containing them or their mixtures, suitable for the treatment treatment/mitigation/regulation of diabetes.
- compositions containing compounds of general formula (I), their tautomeric forms, their pharmaceutically acceptable salts, solvates and their mixtures having pharmaceutically acceptable carriers, solvents, diluents, excipients and other media normally employed in their manufacture are provided.
- Aib ⁇ -Aminoisobutyric acid
- ACN or MeCN Acetonitrile
- Bip Biphenylalanine residue
- Bip(2-CN) 2-nitrile biphenyl residue
- Bip(2-Ipr) 2-Isopropyl biphenyl residue
- DIPCDI Di-isopropylcarbodiimide
- DIPEA Diisopropylethylamine
- Et Ethyl
- Et 2 O Diethyl ether
- Fmoc Fluorenylmethoxycarbonyl
- HOAT 7-Aza-hydroxybenzotriazole
- HBTU 2-(lH-benzotriazole-l-yl)-l,l,3,3-tetramethyl aminium hexafluorophosphate
- PyBOP Benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate
- TrPh 4-phenyl-biphenylalanine residue
- TIPS Triisopropylsilane
- TFA frifluoroacetic acid
- TBTU 2-(lH-benzotriazole-l-yl)-l,l,3,3-tetramethylaminium tetrafluoroborate
- Trt Trityl group
- synthetic GLP-I analog peptides / peptidomimetics which have the structural formula (I), wherein, A represents -NH-R 1 , wherein R 1 represents hydrogen, groups selected from linear or branched (C 1 -C 15 ) alkyl chain, such as methyl, ethyl, propyl, isopropyl, n-butyl, iso- butyl, t-butyl, pentyl, isopentyl, hexyl, heptyl, octyl, decyl groups and the like, an amino acid or peptide containing one, two or three natural amino acid residues, R 3 -CO- group, such as (2-Hydroxy-phenyl)-acetyl group and the like, R 3 O-C(O)- group, such as Fmoc group and the like, a sulfonyl group of formula R 3 -SO 2 -,
- R 4 represents H, optionally substituted groups selected from linear or branched (Ci-C 10 ) alkyl group such as methyl, ethyl, propyl, isopropyl, n-butyl, isorbutyl, t-butyl, pentyl, isopentyl, hexyl, heptyl, octyl, decyl groups and the like, aryl groups selected from phenyl, napthyl, indanyl, fluorenyl, biphenyl and the like; with the proviso that i) when S 1 -Y-S 2 represents a bond, X 1
- HAEGTFTSDVSS (Seq ID 6), HAEGTFTSDVSSY (Seq ID 7), HAEGTFTSDVSSYL (Seq DD 8), HAEGTFTSDVS SYLE (Seq ID 9),
- HAEGTFTSDVSSYLEG (Seq ID 10), HAEGTFTSDVS SYLEGQ (Seq ID 11),
- HAEGTFTSDVSSYLEGQA (Seq JD 12), HAEGTFTSDVS SYLEGQAA (Seq ID 13), HAEGTFTSDVSSYLEGQAAK (Seq TD 14), HAEGTFTSDVS S YLEGQAAKE (Seq ID 15), HAEGTFTSDVSSYLEGQAAKEF (Seq ID 16),
- HAEGTFTSDVSSYLEGQAAKEFI (Seq ID 17), with the further option that one or more of these amino acids may be replaced by unnatural amino acids, and X 2 is selected from the following amino acid sequences GPSSGAPPPS (Seq ID 18)or KELEKLL (Seq ID 19)or GPPS or (Seq ID 20) VKGR (Seq ID 21); ii) and when SpY-S 2 does not represent a bond X 1 is selected from the following amino acid sequences
- HA (Seq ID 22), HAE (Seq ID 23), HAEG (Seq ID 24), HAEGT (Seq ID 25), HAEGTF (Seq ID 26), HAEGTFT (Seq ID 27), HAEGTFTS (Seq ID 28), HAEGTFTSD (Seq ID 29) 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 (Seq ID 18) or KELEKLL (Seq ID 19)or GPPS (Seq ID 20)or VKGR (Seq ID 21)or a dipeptide, selected from combination of two amino acids, consisting of natural or unnatural amino acids, having a side chain containing an arylalkyl or heteroarylalkyl moieties selected from benzyl, napthylmethyl, pyridylmethyl, thienylmethyl, furylmethyl, imidazolylmethyl, isooxazo
- the dipeptide sequence may comprise of one or more amino acids selected from Bip, Bip(2-Me), Bip(2-Et), Bip(2-Ipr), Bip(2-CN), Bip(2'-Et-4'-OMe), B ip(4' -fluoro), Bip(4'-Phenyl), 2-(9,10-Dihydro-phenanthrenyl]- Ala, 2-(Phenanthrenyl)-Ala, 4-(2-Naphthyl)-Phe, 4-(l-Naphthyl)-Phe, 2-Fluorenyl-Ala, 4-dibenzofuran-Phe, 4-dibenzothiophene-Phe, 4-(2 ' -methylphenyl)-3 -pyridylalanine;
- the term 'natural amino acids' indicates all those twenty amino acids, which are present in nature.
- 'unnatural amino acids' or 'non-natural amino acids' represents either replacement of L-amino acids with corresponding D-amino acids such as replacement of L-AIa with D-AIa or L-Pro with D-Pro and the like or suitable modifications of the L or D amino acids, amino alkyl acids, either by
- Such 'unnatural amino acids' or 'non-natural amino acids' may be represented generally by the following structure:
- R 5 wherein Rs is selected from H, F, (C 1 -Cs) alkyl, 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 of R 7 and R 8 is independently selected from H, (C 1 -C 3 ) alkyl, such as methyl and ethyl or halogen atom, preferably fluorine atom; R 9 represents groups, selected from (C 1 -Cs) alkyl, aryl or heteroryl moieties selected from phenyl, napthyl, pyridyl, thienyl, furyl, imidazolyl, isooxazolyl, quinolyl, benzofuranyl, benzothienyl, indolinyl, indolyl, dibenzofuranyl, dibenzothienyl, benzodihydrofuranyl, benzodihydrothienyl, thien
- the suitable substituents include, but are not limited to the following radicals, alone or in combination with other radicals - 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 its derivatives such as esters and amides;
- alkyl used herein, either alone or in combination with other radicals, denotes a linear or branched radical containing one to ten carbons, such as methyl, ethyl, n-propyl, iy ⁇ -propyl, «-butyl, sec-butyl, tert-butyl, amyl, ?-amyi, «-pentyl, n- hexyl, wo-hexyl, heptyl, octyl, decyl and the like.
- cycloalkyl used herein, either alone or in combination with other radicals, denotes a radical containing three to seven carbons, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.
- aryl or "aromatic” used herein, either alone or in combination with other radicals, denotes an aromatic system containing one, two or three rings wherein such rings may be attached together in a pendant manner or may be fused, such as phenyl, naphthyl, tetrahydronaphthyl, indane, biphenyl, and the like.
- arylalkyl denotes an alkyl group, as defined above, attached to an aryl, such as benzyl, phenylethyl, naphthylmethyl, and the like.
- aryloxy denotes an aryl radical, as defined above, attached to an alkoxy group, such as phenoxy, naphthyloxy and the like, which may be substituted.
- aralkoxy denotes an arylalkyl moiety, as defined above, such as benzyloxy, phenethyloxy, naphthylmethyloxy, phenylpropyloxy, and the like, which may be substituted.
- heteroaryl or “heteroaromatic” used herein, either alone or in combination with other radicals, denotes an aromatic system containing one, two or three rings wherein such rings may be attached together in a pendant manner or may be fused containing one or more hetero atoms selected from O, N or S, such as pyridyl, thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, isothiazolyl, imidazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzothienyl, indolinyl, indolyl, azaindolyl, azaindolinyl, benzodihydrofuranyl, benzodihydrothienyl, pyrazolopyrimidinyl, pyr
- heteroaryl used herein, either alone or in combination with other radicals, denotes a heteroaryl group, as defined above, attached to a straight or branched saturated carbon chain containing 1 to 6 carbons, such as (2-furyl)methyl, (3- furyl)methyl, (2-thienyl)methyl, (3-thienyl)methyl, (2-pyridyl)methyl, 1 -methyl- 1 -(2- pyrimidyl)ethyl and the like.
- heteroaryloxy denotes heteroaryl, heteroarylalkyl, groups respectively, as defined above, attached to an oxygen atom.
- acyl used herein, either alone or in combination with other radicals, denotes a radical containing one to eight carbons such as formyl, acetyl, propanoyl, butanoyl, wo-butanoyl, pentanoyl, hexanoyl, heptanoyl, benzoyl and the like, which may be substituted.
- carboxylic acid used herein, alone or in combination with other radicals, denotes a -COOH group, and includes derivatives of carboxylic acid such as esters and amides.
- ester used herein, alone or in combination with other radicals, denotes -COO- group, and includes carboxylic acid derivatives, where the ester moieties are alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl, and the like, which may be substituted.
- the term 'amino acid' as employed herein alone or as part of another group includes, without limitation, an amino group and a carboxyl group linked to the same carbon, referred to as ' ⁇ ' carbon.
- the absolute 'S' configuration at the ' ⁇ ' carbon is commonly referred to as the 'L' or natural configuration.
- the 'R' configuration at the ' ⁇ ' carbon is commonly referred to as the 'D' amino acid.
- the amino acids are GIy or ⁇ ib and are not chiral.
- 'receptor modulator' refers to a compound that acts at the GLP-I receptor to alter its ability to regulate downstream signaling events.
- Example of receptor modulators includes agonist, partial agonist, inverse agonist, allosteric potentiators.
- the isolated peptidomimetics are a 3-30 mer and such peptide bind to and activates the GLP-I receptor.
- the synthetic isolated peptidomimetics described herein possess the ability to mimic the biological activity of GLP-I peptide, with preference for agonist activity at GLP-IR.
- These synthetic peptidomimetics GLP- 1 mimetic exhibit desirable in-vivo properties, thus making them ideal therapeutic candidates for oral or parenteral administration.
- the present invention provides for compounds of formula (I) pharmaceutical compositions employing such compounds either alone or in combination and for methods of using such compounds.
- the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), alone or in combination(s), with a pharmaceutically acceptable carrier.
- a method for treating or delaying the progression or onset of diabetes especially type II diabetes, including complications of diabetes, including retinopathy, neuropathy, nephropathy and delayed wound healing and related diseases such as insulin resistance (impaired glucose homeostasis), hyperglycemia, hyperinsulinemia, elevated blood levels of fatty acids or glycerol, obesity, hyperlipidemia including hypertriglyceridemia, syndrome X, atherosclerosis and hypertension, wherein a therapeutically effective amount of a compound of formula (I) or their combination(s) are administered to a mammal, example, human, a patient in need of treatment.
- Several synthetic routes can be employed to prepare the compounds of the present invention well known to one skilled in the art of peptide synthesis.
- the compounds of formula (I), where all symbols are as defined earlier can be synthesized using the methods described below, together with conventional techniques known to those skilled in the art of peptide synthesis, or variations thereon as appreciated by those skilled in the art. Referred methods include, but not limited to those described below.
- the peptidomimetics thereof described herein may be produced by chemical synthesis using suitable variations of various solid-phase techniques generally known such as those described in G. Barany & R. B. Merrifield, "The peptides: Analysis, synthesis, Biology”; Volume 2- “Special methods in peptide synthesis, Part A", pp. 3- 284, E. Gross & J. Meienhofer, Eds., Academic Press, New York, 1980; and in J. M. Stewart and J. D. Young, "Solid-phase peptide synthesis” 2nd Ed., Pierce chemical Co., Rockford, II, 1984.
- the preferred strategy for preparing the peptidomimetics of this invention is based on the use of Fmoc-based SPPS approach, wherein Fmoc (9-Fluorenyl-methyl- methyloxycarbonyl) group is used for temporary protection of the ⁇ -amino group, in combination with the acid labile protecting groups, such as t-butyloxy carbonyl (Boc), tert-butyl (Bu 4 ), Trityl (Trt) group for temporary protection of the amino acid side chains (see for example E. Atherton & R.C.
- the peptidomimetics can be synthesized in a stepwise manner on an insoluble polymer support (resin), starting form the C-terminus of the peptide.
- the synthesis is initiated by appending the C-terminal amino acid of the peptide to the resin through formation of an amide, ester or ether linkage. This allows the eventual release of the resulting peptide as a C-terminal amide, carboxylic acid or alcohol, respectively.
- the C-terminal amino acid and all other amino acids used in the synthesis are required to have their ⁇ -amino groups and side chain functionalities (if present) differentially protected (orthogonal protection), such that the ⁇ -amino protecting group may be selectively removed during the synthesis, using suitable base such as 20% piperidine solution, without any premature cleavage of peptide from resin or deprotection of side chain protecting groups, usually protected with the acid labile protecting groups.
- suitable base such as 20% piperidine solution
- the coupling of an amino acid is performed by activation of its carboxyl group as an active ester and reaction thereof with unblocked ⁇ -amino group of the N-terminal amino acid appended to the resin.
- peptidyl-resin was washed with the excess of solvents, such as DMF, DCM and diethyl ether.
- solvents such as DMF, DCM and diethyl ether.
- the sequence of ⁇ -amino group deprotection and coupling is repeated until the desired peptide sequence is assembled.
- the peptide is then cleaved from the resin with concomitant deprotection of the side chain functionalities, using an appropriate cleavage mixture, usually in the presence of appropriate scavengers to limit side reactions.
- the resulting peptide is finally purified by reverse phase HPLC.
- the synthesis of the peptidyl-resins required as precursors to the final peptides utilizes commercially available cross-linked polystyrene polymer resins (Novabiochem, San Diego, CA).
- Preferred for use in this invention are Fmoc-PAL-PEG-PS resin, 4-(2', 4'-dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetyl-/7-methyl benzhydrylamine resin (Fmoc-Rink amide MBHA resin), 2-chloro-Trityl-chloride resin or p- benzyloxybenzyl alcohol resin (HMP resin) to which the C-terminal amino acid may or may not be already attached.
- Fmoc-PAL-PEG-PS resin 4-(2', 4'-dimethoxyphenyl-Fmoc-aminomethyl)-phenoxyacetyl-/7-methyl benzhydrylamine resin
- the C-terminal amino acid is not attached, its attachment may be achieved by HOBt active ester of the Fmoc-protected amino acid formed by its reaction with DIPCDI.
- HOBt active ester of the Fmoc-protected amino acid formed by its reaction with DIPCDI.
- 2-Chloro-trityl resin coupling of first Fmoc-protected amino acid was achieved, using DIPEA.
- N- terminal protection of peptidyl resin was selectively deprotected using a solution of 10- 20 % piperidine solution. After every coupling and deprotection, excess of amino acids and coupling reagents was removed by washed with a DMF, DCM and ether.
- Coupling of the subsequent amino acids can be accomplished using HOBt or HOAT active esters produced from DIPCDI/ HOBt or DIPCDI/HOAT, respectively.
- HOBt or HOAT active esters produced from DIPCDI/ HOBt or DIPCDI/HOAT, respectively.
- complete coupling can be achieved using a combination of highly efficient coupling agents such as HBTU, PyBOP or TBTU, with additives such as DIPEA.
- Peptide + Resin The synthesis of the peptide analogs described herein can be carried out by using batchwise or continuos flow peptide synthesis apparatus.
- the non-natural noncommercial amino acids present at different position were incorporated into the peptide chain, using one or more methods known in the art.
- a Fmoc-protected non-natural amino acid was prepared in solution, using appropriate literature procedures.
- the Fmoc-protected Bip analogs, described above were prepared using modified Suzuki cross coupling method, as known in literature (for e.g. Tetrahedron Letter 58, 9633-9695, 2002).
- the Fmoc-protected ⁇ -methylated amino acids were prepared using asymmetric Strecker synthesis, as described for e.g. in Org. Letters 3(8), 1121-1124, 2001.
- the Fmoc-protected N-methylated amino acids were prepared using a literature method as described in for e.g. JOC, 2005, 70, 6918-6920.
- the resulting derivative was then used in the step-wise synthesis of the peptide.
- the required non-natural amino acid was built on the resin directly using synthetic organic chemistry procedures and a linear peptide chain were build.
- the peptide-resin precursors for their respective peptidomimetics may be cleaved and deprotected using suitable variations of any of the standard cleavage procedures described in the literature (see, for example, D. S. King et al. Int. J. peptide Protein res. 36, 1990, 255 - 266).
- a preferred method for use in this invention is the use of TFA cleavage mixture, in the presence of water and TIPS as scavengers.
- the peptidyl-resin was incubated in TFA / Water /TIPS (94:3:3; V: V: V; 10 ml / 100 mg of peptidyl resin) for 1.5-2 hrs at room temperature.
- the cleaved resin is then filtered off, the TFA solution is concentrated or dried under reduced pressure.
- the resulting crude peptide is either precipitated or washed with Et 2 O or is re-dissolved directly into 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 solution of crude peptide is injected into a semi-Prep column (Luna lO ⁇ ; C 18 ; 100 A ° ), dimension 250 X 50 mm and eluted with a linear gradient of ACN in water, both buffered with 0.1 % TFA, using a flow rate of 15 -50 ml /min with effluent monitoring by PDA detector at 220 nm.
- the structures of the purified peptidomimetics can be confirmed by Electrospray Mass Spectroscopy (ES-MS) analysis.
- peptide prepared were isolated as trifluoro-acetate salt, with TFA as a counter ion, after the Prep-HPLC purification.
- some peptides were subjected for desalting, by passing through a suitable ion exchange resin bed, preferably through anion-exchange resin Dowex SBR P(Cl) or an equivalent basic anion-exchange resin.
- TFA counter ions were replaced with acetate ions, by passing through suitable ion-exchange resin, eluted with dilute acetic acid solution.
- selected peptides, with the acetate salt was treated with 4 M HCl. The resulting solution was filtered through a membrane filter (0.2 ⁇ m) and subsequently
- the present invention provides a method of making a peptidomimetics that mimics the activity of an endogenous polypeptide GLP-IR agonist.
- the polypeptide receptor agonist is GLP-I.
- novel compounds of the present invention can be formulated into suitable pharmaceutically acceptable compositions by combining with suitable excipients as are well known.
- the pharmaceutical composition is provided by employing conventional techniques.
- the composition is in unit dosage form containing an effective amount of the active component, that is, the compounds of formula (I) either alone or combination, according to this invention.
- the quantity of active component, that is, the compounds of formula (I) according to this invention, in the pharmaceutical composition and unit dosage form thereof may be varied or adjusted widely depending upon the particular application method, the potency of the particular compound and the desired concentration.
- the daily oral dosage of the active ingredient when used for the indicated effects, will range between about 0.001 to 1000 mg/kg of body weight, preferably between about 0.01 to 100 mg/kg of body weight per day and most preferably between about 0.6 to 20 mg/kg/day.
- Washed resin was incubated in freshly distilled DMF (1 ml / 100 mg of resin), under nitrogen atmosphere for 5 minutes.
- a 0.5 M solution of first Fmoc- protected amino acid (1-3 eq.), pre-activated with HOBt (1-3 eq.) and DIPCDI (1-2 eq.) in DMF was added to the resin, and the resin was then shaken for 1-3 hrs, under nitrogen atmosphere. Coupling completion was monitored using a qualitative ninhydrin test. After the coupling of first amino acid, the resin was washed with DMF, DCM and Diethyl ether (50 ml X 4).
- the Fmoc- protection on first amino acid, coupled with resin was deprotected, using a 10-20% piperidine solution, followed by the coupling the Fmoc-protected second amino acid, using a suitable coupling agents, and as described above.
- the repeated cycles of deprotection, washing, coupling and washing were performed until the desired peptide chain was assembled on resin, as per general scheme above.
- the desired peptidomimetics were cleaved and deprotected from their respective peptidyl-resins by treatment with TFA cleavage mixture as follows.
- a solution of TFA / Water / Triisopropylsilane (95: 2.5: 2.5) (10 ml / 100 mg of peptidyl- resin) was added to peptidyl-resins and the mixture was kept at room temperature with occasional starring.
- the resin was filtered, washed with a cleavage mixture and the combined filtrate was evaporated to dryness. Residue obtained was dissolved in 10 ml of water and the aqueous layer was extracted 3 times with ether (20 ml each) and finally the aqueous layer was freeze-dried.
- Crude peptide obtained after freeze-drying was purified by preparative HPLC as f ⁇ ll ⁇ ws:
- Preparative HPLC was carried out on a Shimadzu LC-8A liquid chromatograph.
- a solution of crude peptide dissolved in DMF or water was injected into a semi-Prep column (Luna lO ⁇ ; C 18 ; 100 A 0 ), dimension 250 X 50 mm and eluted with a linear gradient of ACN in water, both buffered with 0.1 % TFA, using a flow rate of 15 -50 ml / min, with effluent monitoring by PDA detector at 220 ran.
- a typical gradient of 20 % to 70 % of water- ACN mixture, buffered with 0.1 % TFA was used, over a period of 50 minutes, with 1% gradient change per minute.
- the desired product eluted were collected in a single 10-20 ml fraction and pure peptidomimetics were obtained as amorphous white powders by lyophilisation of respective HPLC fractions.
- HPLC analysis of the purified peptidomimetics were collected in
- each peptide was analyzed by analytical RP-HPLC on a Shimadzu LC-IOAD analytical HPLC system.
- analytical HPLC for analytic HPLC analysis of peptidomimetics, Luna 5 ⁇ ; C 18 ; 100 A ° , dimension 250 X 4.6 mm column was used, with a linear gradient of 0.1% TFA and ACN buffer and the acquisition of chromatogram was carried out at 220 nm, using a PDA detector. Characterization by Mass Spectrometry Each peptide was characterized by electrospray ionisation mass spectrometry
- ESI-MS either in flow injection or LC/MS mode.
- Triple quadrupole mass spectrometers API-3000 (MDS-SCIES, Canada) was used in all analyses in positive and negative ion electrospray mode. Full scan data was acquired over the mass range of quadrupole, operated at unit resolution. In all cases, the experimentally measured molecular weight was within 0.5 Daltons of the calculated monoisotopic molecular weight. Quantification of the mass chromatogram was done using Analyst 1.4.1 software.
- the peptidomimetics prepared as described above were tested for GLP-I agonist activity in vitro using the cAMP cell-based assay described below.
- the GLP-I mimetic peptide analog stimulated cAMP production in a dose response manner and the corresponding EC 50 value were determined for some of the selected peptidomimetics, which are active in vitro at 10 to 100 nM range.
- the ECs 0 value of EX-4 was used as a positive control. Cyclic AMP determination
- the GLP-I receptor is a G-protein coupled receptor.
- GLP-I (7-36)-amide the biologically active form, binds to the GLP-I receptor and through signal transduction causes activation of adenylate cyclase and raises intracellular cAMP levels.
- agonism of peptide compounds in stimulating the GLP-I receptor adenyl cyclase activity was monitored by assaying for cellular cAMP levels.
- cAMP assay Stably transfected CHO/HGLP1R cells were assayed for cAMP generation in a semi high throughput platform using DiscoverX cAMP kit with Exendin-4 as a positive control.
- NCEs The activity of NCEs was determined as % Exendin-4 activity at 0.0 l ⁇ M concentration.
- the positive compounds were further validated for cAMP generation using indirect cAMP ELISA kit (R & D systems) The activity of the compounds was expressed as fmol cAMP/ ⁇ g of protein.
- EC5 0 values of some of the representative compounds (I to IV) are shown in Figure 1. Demonstration of in vivo efficacy of compounds:
- the in- vivo glucose lowering properties of some of the representative compounds in animal models is described below.
- This test was used to examine in vivo efficacy of compounds of the present invention on blood glucose at hyperglycemia.
- the intra peritoneal glucose tolerance test (IPGTT) was performed in overnight fasted Swiss Albino Mice (SAM), weighing 25-30 g. Mice were given glucose load of 1.5g/ Kg/ 10 ml and blood was collected at different time intervals, via retroorbital plexus.
- Test compounds (peptidomimetics) were dissolved in an appropriate vehicle at a concentration in nmol/ ml equivalent to the dose that was to be administered in nmol / kg, so that each mouse would receive the same volume / weight of dosing solution.
- AUC Area under the Curve
- BCAUC Base line corrected area under the curve
- the present invention provides novel GLP-I peptide mimics, with a preference for mimicking GLP-I, such that the compounds of the present invention have agonist activity for the GLP-I receptor. Further, many of the GLP peptide mimics of the present invention exhibit increased stability to proteolytic cleavage as compared to GLP-I native sequences.
- the compounds of the present invention can be administered to mammals, preferably humans, for the treatment of a variety of conditions and disorders, including, but not limited to, treating or delaying the progression or onset of diabetes (preferably type II, impaired glucose tolerance, insulin resistance and diabetic complications, such as nephropathy, retinopathy, neuropathy and cataracts), hyperglycemia, hyperinsulinemia, hypercholesterolemia, elevated blood levels of free fatty acids or glycerol, hyperlipidemia, hypertriglyceridemia, obesity, wound healing, tissue ischemia, atherosclerosis, hypertension, intestinal diseases (such as necrotizing enteritis, microvillus inclusion disease or celic disease).
- the compound of the present invention may also be utilized to increase the blood levels of high density lipoprotein (HDL).
- HDL high density lipoprotein
- the conditions, diseases collectively referenced to as 'Syndrome X' or metabolic syndrome as detailed in Johannsson J., Clin. Endocrinol. Metab., 82, 727- 34,1997, may be treated employing the compounds of the invention.
- the compounds of the present invention may optionally be used in combination with suitable DPP-IV inhibitors for the treatment of some of the above disease states either by administering the compounds sequentially or as a formulation containing the compounds of the present invention along with a suitable DPP-IV inhibitors.
- the compounds of the present invention showed good glucose serum- lowering activity in the experimental animals used. These compounds are used for the testing/ prophylaxis of diseases caused by hyperinsulinaemia, hyperglycemia such as NIDDM, metabolic disorders and obesity since such diseases are inter-linked to each other.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Diabetes (AREA)
- Endocrinology (AREA)
- Obesity (AREA)
- Hematology (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Cardiology (AREA)
- Heart & Thoracic Surgery (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Urology & Nephrology (AREA)
- Ophthalmology & Optometry (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Vascular Medicine (AREA)
- Child & Adolescent Psychology (AREA)
- Dermatology (AREA)
- Emergency Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
IN558MU2005 | 2005-05-05 | ||
IN645MU2005 | 2005-05-31 | ||
PCT/IN2006/000154 WO2007017892A2 (en) | 2005-05-05 | 2006-05-04 | Novel compounds as glp-i agonists |
Publications (1)
Publication Number | Publication Date |
---|---|
EP1891106A2 true EP1891106A2 (en) | 2008-02-27 |
Family
ID=37727716
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP06809915A Withdrawn EP1891106A2 (en) | 2005-05-05 | 2006-05-04 | Novel compounds as glp-i agonists |
Country Status (12)
Country | Link |
---|---|
EP (1) | EP1891106A2 (no) |
JP (1) | JP2008540402A (no) |
KR (1) | KR20080021636A (no) |
AP (1) | AP2007004227A0 (no) |
AU (1) | AU2006277557A1 (no) |
BR (1) | BRPI0612471A2 (no) |
CA (1) | CA2606894A1 (no) |
EA (1) | EA200702419A1 (no) |
IL (1) | IL187105A0 (no) |
MX (1) | MX2007013655A (no) |
NO (1) | NO20075618L (no) |
WO (1) | WO2007017892A2 (no) |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7534763B2 (en) | 2004-07-02 | 2009-05-19 | Bristol-Myers Squibb Company | Sustained release GLP-1 receptor modulators |
WO2007139589A1 (en) | 2006-05-26 | 2007-12-06 | Bristol-Myers Squibb Company | Sustained release glp-1 receptor modulators |
MX2009003400A (es) * | 2006-10-03 | 2009-04-28 | Cadila Healthcare Ltd | Compuestos antidiabeticos. |
KR101277560B1 (ko) * | 2007-12-11 | 2013-06-25 | 카딜라 핼쓰캐어 리미티드 | 글루카곤 안타고니스트 활성 및 glp-1 아고니스트 활성의 펩티드 유사체 |
CN101903400B (zh) * | 2007-12-11 | 2013-12-18 | 霍夫曼-拉罗奇有限公司 | 使用固相和液相组合技术的促胰岛素肽合成 |
CN102414220A (zh) * | 2009-05-01 | 2012-04-11 | 霍夫曼-拉罗奇有限公司 | 使用固相和液相组合技术的促胰岛素肽合成 |
CN101891823B (zh) | 2010-06-11 | 2012-10-03 | 北京东方百泰生物科技有限公司 | 一种Exendin-4及其类似物融合蛋白 |
WO2014127120A1 (en) | 2013-02-15 | 2014-08-21 | Mayo Foundation For Medical Education And Research | Insulin secreting polypeptides |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5545618A (en) * | 1990-01-24 | 1996-08-13 | Buckley; Douglas I. | GLP-1 analogs useful for diabetes treatment |
GB0024428D0 (en) * | 2000-10-05 | 2000-11-22 | King S College | Absorption enhancers |
CA2455963C (en) * | 2001-07-31 | 2017-07-04 | The Government Of The United States Of America As Represented By The Secretary, Department Of Health And Human Services | Glp-1, exendin-4, peptide analogs and uses thereof |
US7238671B2 (en) * | 2001-10-18 | 2007-07-03 | Bristol-Myers Squibb Company | Human glucagon-like-peptide-1 mimics and their use in the treatment of diabetes and related conditions |
EP1585959A4 (en) * | 2002-01-08 | 2007-11-14 | Lilly Co Eli | PEPTIDE-1 ANALOGUES OF EXTENSION GLUCAGON TYPE |
CN1961000B (zh) * | 2004-02-11 | 2011-05-04 | 安米林药品公司 | 具有可选择特性的杂合多肽 |
TW200611704A (en) * | 2004-07-02 | 2006-04-16 | Bristol Myers Squibb Co | Human glucagon-like-peptide-1 modulators and their use in the treatment of diabetes and related conditions |
DE102004043153B4 (de) * | 2004-09-03 | 2013-11-21 | Philipps-Universität Marburg | Erfindung betreffend GLP-1 und Exendin |
-
2006
- 2006-05-04 KR KR1020077028300A patent/KR20080021636A/ko not_active Application Discontinuation
- 2006-05-04 CA CA002606894A patent/CA2606894A1/en not_active Abandoned
- 2006-05-04 MX MX2007013655A patent/MX2007013655A/es not_active Application Discontinuation
- 2006-05-04 AP AP2007004227A patent/AP2007004227A0/xx unknown
- 2006-05-04 AU AU2006277557A patent/AU2006277557A1/en not_active Abandoned
- 2006-05-04 WO PCT/IN2006/000154 patent/WO2007017892A2/en active Application Filing
- 2006-05-04 EA EA200702419A patent/EA200702419A1/ru unknown
- 2006-05-04 EP EP06809915A patent/EP1891106A2/en not_active Withdrawn
- 2006-05-04 JP JP2008509578A patent/JP2008540402A/ja not_active Withdrawn
- 2006-05-04 BR BRPI0612471A patent/BRPI0612471A2/pt not_active IP Right Cessation
-
2007
- 2007-11-01 IL IL187105A patent/IL187105A0/en unknown
- 2007-11-06 NO NO20075618A patent/NO20075618L/no not_active Application Discontinuation
Non-Patent Citations (1)
Title |
---|
See references of WO2007017892A2 * |
Also Published As
Publication number | Publication date |
---|---|
NO20075618L (no) | 2008-01-30 |
EA200702419A1 (ru) | 2008-04-28 |
MX2007013655A (es) | 2008-01-24 |
IL187105A0 (en) | 2008-02-09 |
WO2007017892A3 (en) | 2007-09-20 |
KR20080021636A (ko) | 2008-03-07 |
AP2007004227A0 (en) | 2007-12-31 |
WO2007017892A2 (en) | 2007-02-15 |
CA2606894A1 (en) | 2007-02-15 |
BRPI0612471A2 (pt) | 2016-09-06 |
JP2008540402A (ja) | 2008-11-20 |
AU2006277557A1 (en) | 2007-02-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2007323035B2 (en) | Antidiabetic compounds | |
US8883963B2 (en) | Peptidomimetics with glucagon antagonistic and GLP 1 agonistic activities | |
KR101205272B1 (ko) | 아실화된 glp-1 화합물 | |
WO2014049610A2 (en) | Peptides as gip, glp-1 and glucagon receptors triple-agonist | |
WO2007017892A2 (en) | Novel compounds as glp-i agonists | |
KR20070120112A (ko) | 연장형 glp-1 화합물 | |
CN113366014A (zh) | 经修饰的gip肽类似物 | |
US20150141336A1 (en) | Pancreatic Peptide Compounds and Use | |
BR112012029248B1 (pt) | Análogo de glp-1 de fórmula i ou a sua composição, análogo de glp-1 de fórmula viii ou um sal farmaceuticamente aceitável ou a composição do mesmo, composição farmacêutica e uso de um composto | |
US20120264685A1 (en) | Short chain peptidomimetics based orally active glp 1 agonist and glucagon receptor antagonist | |
CN101223189A (zh) | 作为glp-i激动剂的新颖化合物 | |
CN115960258A (zh) | 一类GLP-1/glucagon/Y2受体三重激动剂及其应用 | |
JP2012513981A (ja) | Glp−1アナログおよびその使用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 20071107 |
|
AK | Designated contracting states |
Kind code of ref document: A2 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI SK TR |
|
AX | Request for extension of the european patent |
Extension state: AL BA HR MK YU |
|
17Q | First examination report despatched |
Effective date: 20090204 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 20090616 |