MXPA06008909A - Bicycloester derivative - Google Patents

Abstract

[PROBLEMS]To provide a novel bicycloester derivative having excellent DPP-IV inhibiting activity or a pharmacologically acceptable salt thereof. [MEANS FOR SOLVING PROBLEMS]There is provided a novel bicycloester derivative of the general formula:(1) or a pharmacologically acceptable salt thereof (for example, (2S, 4S)-1-[[N- (4-ethoxycarbonylbicyclo [2.2.2]oct -1-yl)amino]acetyl]-4-fluoropyrrolidine -2-carbonitrile).

Description

DERIVATIVE OF BICICLOESTER FIELD OF THE INVENTION The present invention relates to bicycloester derivatives and pharmaceutically acceptable salts thereof having dipeptidylpeptidase IV (DDP-IV) inhibitory activity and are useful in the prevention and / or treatment of type II diabetes and other diseases involving DPP-IV.

BACKGROUND OF THE INVENTION Dipeptidylpeptidase IV (EC3.4.14.5, referred to as "DPP-IV" or "CD26" below) is a serine protease that specifically hydrolyzes polypeptides having proline or alanine at position 2 at the C-terminus of these amino acid residues, by cleaving dipeptides Xaa-Pro or Xaa-Ala from the N-terminus of the polypeptides (Xaa can be any amino acid). A biological function of DPP-IV is the inactivation of glucagon-like peptide 1 (GLP-1) by hydrolyzing the N-terminal His-Ala dipeptide of GLP-1 (Document 1 without patent). GLP-1 inactivated by DPP-IV is thought to act as an antagonist in the receptors GLP-1 also decreases the physiological activity of GLP-1 (Document 2 without patent). GLP-1, a hormone The secreted peptide of the endocrine L cells, found mainly in intestinal epithelium, is known to act on β-cells of the pancreatic Langerhans islets in a glucose-dependent manner, to promote insulin secretion, thus decreasing the glucose level of the blood (Document No. 3 without patent). Having an ability to promote insulin biosynthesis and ß cell growth, GLP-1 is an essential factor for the maintenance of β cells (Documents 5 and 6 without patent). It has been reported that GLP-1 also acts to promote the use of glucose by peripheral tissue and, when administered intravenously, decreases the admission of food and motility of the Gl tract (Documents 7 to 10 without patent). A DPP-IV inhibitor is believed to increase the activity of GLP-1 by suppressing the innate GLP-1 composition. The increased GLP-1 activity stimulates insulin secretion and improves glucose metabolism. For this reason, DPP-IV inhibitors are expected to be useful agents in the prevention and / or treatment of diabetes, in particular type II diabetes (Documents 11 and 12 without patent). The compounds are also expected to be effective in the prevention and / or treatment of other diseases that are caused or worsened by the decrease in glucose metabolism (e.g., diabetic complications, hyperinsulinemia, hyperglycemia, abnormal lipid metabolism and obesity). The roles of DPP-IV have been described in a living body other than the inactivation of GLP-1 and how the enzyme is involved in the attack of various diseases, in many reports as described below. (a) DPP-IV inhibitors and their antibodies, prevent the invasion of HIV into cells. The expression of CD26 is reduced in T cells derived from patients infected with HIV-1 (Document 13 without patent). The Tat proteins of HIV-1 bind to DPP-IV (Document 14 without patent). (b) DPP-IV is involved in immune responses. DPP-IV inhibitors and their antibodies suppress the growth of T cells stimulated by antigens (Document 15 without patent). T cells stimulated by antigens, express an increased level of DPP-IV (Document without patent). DPP-IV is involved in the production of cytokine and other T cell functions (Document 17 without patent). DPP-IV binds to adenosine deaminase (ADA) on the surface of T cells (Document 18 without patent). (c) The expression of DPP-IV is increased in the skin fibroblasts of patients with rheumatoid arthritis, psoriasis, and lichen planus (Document 19 without patent). (d) High DPP-IV activity is observed in patients with benign prostatic hypertrophy and homogenate of prostatic tissue (Document 20 without patent). DPP-IV in the lung endothelium acts as an adhesive molecule for prostate cancer and lung metastatic breast cancer in rats (Document 21 without patent). (e) The DPP-IV defective variant of rats F344, has lower blood pressure than the native type F344 rats. DPP-IV interacts with a protein that plays a crucial role in the reabsorption of sodium by the kidney (Documents 1 and 2 of the patent). (f) The inhibition of DPP-IV activity offers an effective procedure for the prevention and / or treatment of myelosuppressive diseases, while the agents that activate DPP-IV are expected to serve as drugs to increase the cell count of blood cells targets and / or to treat infectious diseases (Patent Document 3). These observations indicate that DPP-IV inhibitors can be useful agents in the prevention and / or treatment of diabetes (in particular, type II diabetes) and / or other diseases that complicate diabetes involving DPP-IV. For example, DPP-IV inhibitors they are expected to be useful in the prevention and / or treatment of AIDS after infection with HIV-1, rejection after organ / tissue transplantation, multiple sclerosis, rheumatoid arthritis, inflammation, allergies, osteoporosis, psoriasis and lichen planus, prostatic hypertrophy benign, metastasis of lung and prostate cancers, hypertension and infectious diseases. DPP-IV inhibitors are also expected to be useful in facilitating diuresis, decreased myelosuppression and decreased white blood cell count. Among existing DPP-IV inhibitors are pyrrolidine derivatives described in Documents 4 to 11 of the Patent, heterocyclic derivatives are described in Documents 12 and 13 of the Patent, and β-amino acid derivatives are described in Documents 14 and 15 of the Patent. Patent Document 16, a United States patent, describes a simple bicyclo [2.2.2] octane derivative having inhibitory activity for DPP-IV. This compound, however, is completely different from the compounds of the present invention in its structure and mechanism for DPP-IV inhibition. Patent Document 17 mentions a bicyclic derivative, structurally similar to the compounds of the present invention. However, there is no description in this Literature that refers to any of the compounds of the present invention, nor has any example of the compounds been presented. None of the previously described DPP-IV inhibitors are practically sufficient in terms of the DPP-IV inhibitory activity, selectivity for DPP-IV, stability, toxicity and biological kinetics. Thus, there is a constant need for effective DPP-IV inhibitors.

[Document 1 without patent] American Journal of Physiology, Vol. 271 (1996): ppE458-E464. [Document 2 without Patent] European Journal of Pharmacology, Vol. 318 (1996: pp429-435. [Document 3 without Patent] European Journal Clinical Investigation, Vol, 22 (1992): pl54. [Document 4 without a patent] Lancet, Vol. 2 (1987): pl300. [Document 5 without Patent] Endocrinology, Vol. 42 (1992): p85ß. [Document 6 without Patent] Diabetologia, Vol. 42 (1999): p 856. [Paper 7 without Patent] Endocrinology, Vol. 135 (1994): p2070. [Document 8 without Patent], Vol. 37 (1994): pll63. [Document 9 without Patent] Digestion, Vol. 54 (1993): p 392.

[Document 10 without Patent] Dig. Dis. Sci., Vol 43 (1998): plll3.

[Document 11 without Patent] Diabetes, Vol. 47 (1998): pplßß3-1670. [Paper 12 without Patent] Diabetologia, Vol. 42 (1999): ppl342-1331. [Document 13 without Patent] Journal of Immunology, Vol. 149 (1992): p3037. [Document 14 without Patent] Journal of Immunology, Vol. 150 (1993): p2544. [Paper 15 without Patent] Biological Chemistry (19991): p305. [Document 16 without Patent] Scandinavian Journal of Immunology, Vol. 33 (1991): p737. [Document 17 without Patent] Scandivavian Journal of Immunology, Vol. 29 (1989): pl27. [Paper 18 without Patent] Science, Vol. 261 (1993): p466.

[Document 19 without Patent] Journal of Cellular Physiology, Vol. 151 (1992): p378. [Document 20 without Patent] European Journal of Clinical Chemistry and Clinical Biochemistry, Vol. 30 (1992): p333. [Document 21 without Patent] Journal of Cellular Physiology, Vol. 121 (1993): pl423. [Patent Document 1] WO 03/015775 Booklet [Patent Document 2] WO 03/017936 Booklet [Patent Document 3] WO 03/080633 Booklet [Patent Document 4] WO 95/15309 Brochure [Patent Document 5] WO 98/19998 Booklet [Patent Document 6] WO 00/34241 Booklet [Patent Document 7] WO 02/14271 Booklet [Patent Document 8] WO 02/30890 Booklet [Patent Document 9] WO 02/38541 Booklet [Patent Document 10] WO 03/002553 Booklet [Patent Document 11] US 02/0193390 Publication [Patent Document 12] WO 02/062764 Booklet [Patent Document 13] WO 03/004496 Booklet [ Patent Document 14] WO 03/000180 Booklet [Patent Document 15] WO 03/004498 Booklet [Patent Document 16] US 02/0193390 Publication [Patent Document 17] WO 02/38541 Brochure BRIEF DESCRIPTION OF THE FIGURES Fig. 1 is a graph showing the results of Test Example 3.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a novel compound having high inhibitory activity of DPP-IV, as well as pharmaceutically acceptable salts thereof. It is another object of the present invention to provide a pharmaceutical composition containing the new compound having high activity inhibitor of DPP-IV or a pharmaceutically acceptable salt thereof. It is still another object of the present invention to provide a prophylactic and / or therapeutic agent for diabetes and associated with complications, as well as a prophylactic and / or therapeutic agent for diseases involving DPP-IV. In accordance with the present invention, there is provided a novel bicycloester derivative having high inhibitory activity for DPP-IV, and pharmaceutically acceptable salts thereof. Also provided is a pharmaceutical composition containing the novel bicycloester derivatives having high inhibitory activity for DPP-IV, or a pharmaceutically acceptable salt thereof. In addition, a prophylactic and / or therapeutic agent for diabetes and associated complications, and a prophylactic and / or therapeutic agent for diseases involving DPP-IV are provided. Thus, the present invention relates to the following: 1) A bicycloester derivative represented by the following general formula (1): wherein R1 is a substituted or unsubstituted Ci to C6 alkyl group, substituted or unsubstituted C3 cycloalkyl group, tetrahydropyranyl group, substituted or unsubstituted arylmethyl group, substituted or unsubstituted arylethyl group, substituted or unsubstituted aromatic hydrocarbon group, substituted aromatic heterocyclic ring or unsubstituted, or substituted or unsubstituted aliphatic heterocyclic ring; X is CH2, CHF, CF2, CHOH, S, or O; n is 1, 2 or 3, or a pharmaceutically acceptable salt thereof. 2) An intermediate in the production of the bicycloester derivative of 1) above, represented by the following general formula (2): (2) wherein R 1 is a substituted or unsubstituted C 1 to C 4 alkyl group, substituted or unsubstituted C 3 to C 6 cycloalkyl group, tetrahydropyranyl group, substituted or unsubstituted arylmethyl group, substituted or unsubstituted arylethyl group, substituted or unsubstituted aromatic hydrocarbon group, substituted aromatic heterocyclic ring or unsubstituted or substituted or unsubstituted aliphatic heterocyclic ring; X is CH2, CHF, CF2, CHOH, S or O; n is 1, 2 or 3; and p1 is a protecting group of an amino group. 3) A pharmaceutical product containing as an active ingredient, the bicycloester derivative of 1) above or a pharmaceutically acceptable salt thereof. 4) A DPP-IV inhibitor containing as an active ingredient the bicycloester derivative of 1) above or a pharmaceutically acceptable salt thereof. 5) A therapeutic agent for a disease involving DPP-IV containing as an active ingredient the bicycloester derivative of 1) above a pharmaceutically acceptable salt thereof. 6) The therapeutic agent according to 5) above, wherein the disease involving DPP-IV is diabetes or an associated complication. The term "substituted or unsubstituted C 1 to C 1 alkyl group" as used herein refers to a Ci to C 1 alkyl group (such as methyl group, cyclopropylmethyl group, ethyl group, propyl group, 1-methylethyl group, group 1 methylpropyl, 2-methylpropyl group, 1-ethylpropyl group, 2-ethylpropyl group, butyl group, t-butyl group and hexyl group) which may have 1 to 5 substituents selected from halogen atom, hydroxy group, cyano group, alkoxy group Ci a Cß, substituted or unsubstituted aryloxy group, alkylcarbonyl group Ci a er alkoxycarbonyl group Ci to Ce, alkylthio group Ci to Cß, amino group, Ci to C6 alkylamino group mono- or disubstituted, cyclic amino group of 4 to 9 elements which may contain 1 to 3 heteroatoms, formylamino group, alkylcarbonyl group Ci to Cß, alkoxycarbonylamino group Ci to C6, alkylsulfonylamino group Ci to Ce and substituted or unsubstituted arylsulfonylamino group. The term "substituted or unsubstituted C3 to C3 cycloalkyl group" refers to a C3 to C3 cycloalkyl group (such as cyclopropyl group, cyclobutyl group, cyclopentyl group and cyclohexyl group) which may have 1 to 5 substituents selected from halogen atom, group hydroxy, cyano group, Ci to β alkoxy group, substituted or unsubstituted aryloxy group, Ci to C6 alkylcarbonyl group, Ci to C6 alkoxycarbonyl group, Ci to C6 alkylthio group, amino group, mono- or di-substituted C 1 to C 1 alkylamino group, amino group cyclic from 4 to 9 elements which can contain 1 to 3 heteroatoms, formylamino groups, alkylcarbonylamino group Ci to Cß, alkoxycarbonylamino group Ci to Cg, alkylsulfonylamino group Ci to Cß and substituted or unsubstituted arylsulfonylamino group. The term "substituted or unsubstituted arylmethyl group" refers to an arylmethyl group (such as phenylmethyl group, naphthylmethyl group, pyridylmethyl group, quinolylmethyl group and indolylmethyl group) which may have 1 to 5 substituents selected from halogen atom, substituted or unsubstituted Ci to C6 alkyl group, hydroxy group, cyano group, nitro group, substituted or unsubstituted Ci to C6 alkoxy group, substituted or unsubstituted aryloxy group, alkylcarbonyl group Ci to Ce, group alkoxycarbonyl Ci to C, alkylthio group C to C, amino group, mono- or di-substituted or unsubstituted alkylamino Ci to Cβ group, substituted or unsubstituted arylamino group, cyclic amino group of 4 to 9 elements which may contain 1 to 3 heteroatoms, formylamino group, C1 to C6 alkylcarbonylamino group, Ci to C6 alkoxycarbonylamino group, Ci to C6 alkylsulfonylamino group, and substituted or unsubstituted arylsulfonylamino group. The term "substituted or unsubstituted ethyl group" refers to an arylethyl group (such as 1-phenethyl group, 2-phenethyl group, 1-naphthylethyl group and 2-naphthylethyl group) which may have 1 to 5 substituents selected from halogen atom , substituted or unsubstituted Ci to C6 alkyl group, hydroxy group, cyano group, nitro group, substituted or unsubstituted Ci to C alkoxy group, substituted or unsubstituted aryloxy group, Ci to C6 alkylcarbonyl group, Cx to C6 alkoxycarbonyl group, Ci alkylthio group a C5, amino group, Ci-C6 alkylamino group mono- or di-substituted or unsubstituted, substituted or unsubstituted arylamino group, cyclic amino group of 4 to 9 elements which may contain 1 to 3 heteroatoms, formylamino group, alkylcarbonylamino group Ci to C6, alkoxycarbonylamino group Ci to C6, alkylsulfonylamino group Cx to Cß and substituted or unsubstituted arylsulfonylamino group. The term "substituted or unsubstituted aromatic hydrocarbon group" refers to an aromatic hydrocarbon group (such as benzene ring, naphthalene ring and anthracene ring) which may have 1 to 5 substituents selected from halogen atom, hydroxy group, cyano group , nitro group, alkoxy group Ci to Cg, alkylthio group Ci to C6, amino group, alkylamino group Ci to mono- or di-substituted Ce, cyclic amino group of 4 to 9 elements which may contain 1 to 3 heteroatoms, formylamino group, alkylcarbonylamino group Ci to C6, alkylsulfonylamino group C to C and arylsulfonylamine group substituted or unsubstituted. The term "substituted or unsubstituted aromatic heterocyclic ring" refers to an aromatic heterocyclic ring (including 5- or 6-membered aromatic monocyclic heterocyclic ring or a 9- or 10-membered aromatic fused heterocyclic ring, such as pyridine ring, pyrimidine ring , pyrididazine ring, triazine ring, quinoline ring, naphthylidine ring, quinazoline ring, acridine ring, pyrrolo ring, furan ring, ring thiophene, imidazole ring, pyrazole ring, oxazole ring, isoxazole ring, thiazole ring, indole ring, benzofuran ring, benzothiazole ring, benzimidazole ring and benzoxazole ring, containing 1 to 3 heteroatoms arbitrarily selected from nitrogen, oxygen or sulfur) which can have from 1 to 5 substituents selected from halogen atom, hydroxy group, cyano group, nitro group, Ci to C6 alkoxy group, Cx to C2 alkylthio group, amino group, Ci-alkylamino group a C6 mono- or di-substituted, cyclic amino group of 4 to 9 elements which may contain 1 to 3 heteroatoms, formylamino group, alkylcarbonylamino group Ci to C6, alkylsulfonylamino group Ci to C6 and substituted or unsubstituted arylsulfonylamino group. The term "substituted or unsubstituted aliphatic heterocyclic ring" refers to an aliphatic heterocyclic ring (including 4- to 7-membered aliphatic monocyclic heterocyclic ring or a 9- or 10-membered aliphatic fused heterocyclic ring, such as azetidine ring, pyrrolidine ring , tetrahydrofuran ring, piperidine ring, morpholine ring and piperazine ring, containing 1 to 3 heteroatoms arbitrarily selected from nitrogen, oxygen and sulfur) which may have from 1 to 5 substituents selected from halogen atom, Ci. C6 substituted or unsubstituted, hydroxy group, cyano group, nitro group, substituted or unsubstituted Ci to C6 alkoxy group, CX to C6 alkylthio group, amino group, Ci to C6 alkylamino group or di-substituted or unsubstituted, cyclic amino group of 4 to 9 elements which may contain 1 to 3 heteroatoms, formylamino group, alkylcarbonylamino group Ci to C, alkoxycarbonylamino group Ci to C6, alkylsulfonylamino group C to C6 and substituted or unsubstituted arylsulfonylamino group. The term "substituted or unsubstituted Ci to C6 alkoxy group" refers to a Ci to C6 alkoxy group (such as methoxy group, ethoxy group, butoxy group and hexyloxy group) which may have 1 to 5 selected substituents, and hexyloxy group) which may have 1 to 5 substituents selected from halogen, hydroxy group, cyano group, Ci to C6 alkoxy group, Ci to C6 alkylthio group, amino group, mono- or di-substituted C 1 to C 6 alkylamino group, cyclic amino group from 4 to 9 elements they may contain 1 to 3 heteroatoms, formylamino group, alkylcarbonylamino group Ci to C6, alkylsulfonylamino group Ci to C6 and substituted or unsubstituted arylsulfonylamino group. The term "protecting group for an amino group" refers to t-butoxycarbonyl group. Benzyloxycarbonyl group, allyloxycarbonyl group, methoxycarbonyl group, ethoxycarbonyl group, 2,2,2- trichloroethoxycarbonyl, trifluoroacetyl group, acetyl group, benzyl group and 2, 4,6-trimethoxybenzyl group. The term "halogen atom" refers to fluorine atom, chlorine atom, bromine atom and iodine atom. Preferred examples of the compounds of the compounds of the present invention include (2S, 4S) -l- [[N- (4-ethoxycarbonylbicyclo [2.2.2] oct-1-yl) amino] acetyl] -4-fluoropyrrolidin-2 -carbonitrile, (2S) -1- [[N- (4-ethoxycarbonylbicyclo [2.2.2] oct-1-yl) amino] acetyl] pyrrolidine-2-carbonitrile, (2S) -l- [[N- (4 -t-Butoxycarbonylbicyclo [2.2.2] oct-1-yl) amino] acetyl] pyrrolidine-2-carbonitrile, (2S, 4S) -l - [[N- [4- (2-tetrahydropyranyl) oxycarbonylbicyclo [2.2.2 ] oct-1-yl] amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile, (2S) -l- [[N- [4- (2-tetrahydropyranyl) oxycarbonylbicyclo [2.2.2] oct-1-yl] amino] acetyl] pyrrolidine-2-carbonitrile, (2S, 4S.) -l- [[N- (4-ethoxycarbonylbicyclo [2.2.1] hept-1-yl) amino] acetyl] -4-fluoropyrrolidin-2- carbonitrile, (2S) -1- [[N- (4-ethoxycarbonylbicyclo [2.2.1] hept-1-yl) amino] acetyl] pyrrolidine-2-carbonitrile, (2S, 4S) -l - [[N- ( 4-benzyloxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile, (2S, 4S) -1- [[N- (4-cyclopropylmethyloxycarbonyl bicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile, (2S, 4S) -4-fluoro -l- [[N- (4- (4-trifluoromethyl) benzyloxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] -rrololidin-2-carbonitrile, (2S, 4S) -4-fluoro-l- [ [N- (4-isobutyloxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] pyrrolidine-2-carbonitrile, and (2S, 4S) -4-fluoro-l - [[N- (4-isopropyloxycarbonylbicyclo [ 2.2.2] oct-l-yl) amino] acetyl] pyrrolidine-2-carbonitrile.

ADVANTAGES OF THE INVENTION The new compounds have high inhibitory activity against DPP-IV, of the compounds of the present invention, are useful agents in the prevention and / or treatment of diabetes associated with complications, as well as in the prevention and / or treatment of diseases that involve DPP-IV.

DETAILED DESCRIPTION OF THE INVENTION When the compounds of the present invention form pharmaceutically acceptable salts, examples of such salts include salts with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid or organic acids, such as acetic acid, maleic acid, fumaric acid, succinic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, p-acid toluenesulfonic, benzenesulfonic acid, salicylic acid, stearic acid, palmitic acid and trifluoroacetic acid; metal salts, such as sodium salt, potassium salt, calcium salt, magnesium salt, aluminum salt and zinc salt; ammonium salts, such as ammonium salt and tetramethylammonium salt; organic amino salts, such as morpholine and piperidine; and amino acid salts, such as salts with glycine, lysine, arginine, phenylalanine and proline. The compounds of the present invention represented by the general formula (1) or salts thereof may contain one, two or more chiral centers and thus have multiple optical isomers resulting from these chiral centers. Any of these optical isomers and diastereomers are encompassed by the present invention, as are any mixtures thereof in an arbitrary mixing ratio, including racemic mixtures. When the compounds of the present invention represented by the general formula (1) or salts thereof contain a double bond, they may have Z or E configurations and any of the mixtures of these compounds in an arbitrary mixing ratio are also encompassed by the present invention. Some of these compounds of the present invention represented by the general formula (1) or salts thereof may have tautomers or rotary isomers, all isomers are encompassed by the present invention, as are any of the mixtures thereof in a proportion of arbitrary mixing. The compounds of the present invention represented by the general formula (1) or salts thereof, include intramolecular salts, addition products, solvates and hydrates thereof. The compounds of the present invention represented by the general formula (1) or salts thereof, can be used as a pharmaceutical composition, either individually or in conjunction with one or more pharmaceutically acceptable auxiliary agents. They can be formulated with pharmaceutically acceptable carriers or excipients (such as starch, lactose, calcium phosphate and calcium carbonate), lubricant (such as magnesium stearate, calcium talc stearate and stearic acid), binders (such as starch, crystalline cellulose). , carboxymethylcellulose, gum arabic, polyvinylpyrrolidone and alginic acid), agents disintegrants (such as calcium talc and carboxymethylcellulose) or diluents (such as saline, aqueous solutions of glucose, mannitol or lactose). Using ordinary techniques, the compounds of the present invention represented by the general formula (1) or salts thereof, can be formulated into tablets, capsules, granules, powders, soft granules, ampoules or injections for oral or parenteral administration. The compounds of the present invention represented by the general formula (1) or salts thereof, are generally administered to humans and other animals at a dose of 0.0001 to 1000 mg / kg / day while the dose may vary depending on the type of compounds or salts, route of administration, and the age, body weight and symptoms of the subjects. The compounds of the present invention or salts thereof can be administered in a single daily dose or multiple doses per day. When necessary, the compounds of the present invention represented by the general formula (1) or salts thereof can be used in conjunction with one or more therapeutic agents other than DPP-IV inhibitors. Among such diabetic therapeutic agents for use with the compounds of the present invention or salts thereof, are insulin and its derivatives, GLP-1 and its derivatives and other oral diabetic therapeutic agents. Examples of such diabetic therapeutic agents include diabetic sulfonyl urea therapeutic agents, insulin secretagogues without sulfonylurea, diabetic biguanide therapeutic agents, a-glycosidase inhibitors, glucagon antagonists, GLP-1 agonists, PPAR agonists, β3 agonists, SGLT inhibitors, inhibitors PKC, glucagon synthase kinase 3 (GSK-3) inhibitors, protein tyrosine phosphatase IB (PTP-1B) inhibitors, potassium channel openers, insulin synthesizers, glucose recovery modulators, compounds that modify lipid metabolism and appetite suppressants. Examples of GLP-1 and its derivatives include betatropine and NN-2211. Examples of diabetic sulfonylurea therapeutic agents include tolbutamide, glibenclamide, gliclazide, glimepiride and glipizide. Examples of insulin secretagogues without sulfonylureas include nateglinide, repaglinide, mitiglinide and JTT-608. Examples of diabetic biguanide therapeutic agents include metformin. Examples of a-glycosidase inhibitors include voglibose and miglitol. Examples of PPAR agonists include troglitazone, rosiglitazone, pioglitazone, ciglitizone, KRP-297 (MK-767), isaglitazone, GI-262570, and JTT-501. Examples of β3 agonists include AJ-9677, YM-178 and N-5984. The compounds (1) of the present invention are They can produce by various synthetic techniques. The compounds (1) of the present invention can be isolated or purified by common separation means (such as extraction, recrystallization, distillation and chromatography). The compounds can be obtained in the form of various salts using common techniques or similar techniques (such as neutralization). Representative processes for reducing the compounds of the present invention and salts thereof will now be described.

Process A Step 1 (Process A) In this step, a haloacetic acid derivative of the general formula (4) (wherein Y 1 is Cl or Br, and X is as defined above), is reacted with a bicycloamine derivative of the general formula (3) (wherein R1 and n are as defined above), to obtain a bicyclic ester derivative of the general formula (1) (wherein R1, n and X are as defined above). The reaction was carried out in the presence or absence of one base. The base for use in this reaction may be an inorganic base, such as sodium hydroxide, potassium hydroxide, sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate and cesium carbonate, or an organic base, such as triethylamine, diisopropylethylamine, N, N, N, N-tetramethylethylenediamine, diazabicyclo [5.4.0] -7-undecene, diazabicyclo [4.3.0] -5-nonene, phosphazene base and pentaisopropylguanidine. When it is desired to use a catalyst in the reaction, such a catalyst may be a phase transfer catalyst or an inorganic salt, such as tetrabutylammonium bromide, tetrabutylammonium iodide, benzyltriethyl onium bromide, lithium bromide, lithium iodide, sodium, potassium bromide, potassium iodide, cesium bromide and cesium iodide. The solvent for use in the reaction may be an inert solvent such as acetone, ethanol, toluene, ketonitrile, tetrahydrofuran, dioxane, ethyl ether, t-butyl methyl ether, dimethoxyethane, ethyl acetate, dichloromethane, N, -dimethylformamide, dimethylsulfoxide and N-methyl. -2-pyrrolidine. This reaction proceeds smoothly at 0 to 150 ° C.

Process B (10) Step 1 (Process B) In this step, a haloacetic acid derivative of the general formula (4) (wherein X and Y1 are as defined above), is reacted with a bicycloamine derivative of the general formula (5) (wherein P2 is a protecting group for a carboxyl group, and n is as defined above), to obtain a bicycloester derivative of the general formula (6) (wherein P2, n and X are as defined above). The reaction is carried out in the presence or absence of a base. The base for use in this reaction may be an inorganic base, such as sodium hydroxide, potassium hydroxide, sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate and cesium carbonate, or an organic base, such as triethylamine, diisopropylethylamine, N, N, N, N-tetramethylethylenediamine, diazabicyclo [5.4.0] -7-undene, diazabicyclo [4.3.0] -5-nonene, phosphazene base and pentaisopropylguanidine. When it is desired to use a catalyst in the reaction, the catalyst can be a phase transferase catalyst or an inorganic salt, such as tetrabutylammonium bromide, tetrabutylammonium iodide, benzyltriethylammonium bromide, lithium bromide, lithium iodide, sodium iodide. , potassium bromide, potassium iodide, cesium bromide and cesium iodide. The solvent for use in the reaction may be an inert solvent such as acetone, ethanol, toluene, acetonitrile, tetrahydrofuran, dioxane, ethyl ether, t-butyl methyl ether, dimethoxyethane, ethyl acetate, dichloromethane, N, N-dimethylformamide, dimethylsulfoxide and N- methyl-2-pyrrolidine. This reaction proceeds smoothly at 0 to 150 ° C.

Step 2 (Process B) In this step, the secondary amino group of the bicycloester derivative of the general formula (6) (wherein P2, n and X are as defined above) is protected to give a bicycloester derivative of the general formula (7) in accordance with claim 2 (wherein P1 is a protecting group for an amino group, and P2, n and X are as described above). The protecting group Pl for the secondary amino group can be t-butoxycarbonyl group, benzyloxycarbonyl group or group trifluoroacetyl. Protecting groups can be introduced by known techniques. When P1 is t-butoxycarbonyl group, it can be readily introduced by reacting di-t-butyldicarbonate with the bicycloester derivative of the general formula (6) (wherein P2, n and X are as defined above) in the presence or absence of triethylamine or 4-dimethylaminopyridine. When P1 is benzyloxycarbonyl group, it can be readily introduced by reacting benzyloxycarbonyl chloride with the bicycloester derivative of the general formula (6) (wherein P2, n and X are as defined above) in the presence of triethylamine, diisopropylethylamine or sodium carbonate. potassium. When P1 is trifluoroacetyl group, it can be readily introduced by reacting trifluoroacetic acid anhydride with the bicycloester derivative of the general formula (6) (wherein P2, n and X are as defined above) in the presence of triethylamine or N, N -dimethylaminopyridine.

Step 3 (Process B) In this step, the group P2 protecting the carboxyl group of the bicycloester derivative of the general formula (7) (wherein P2, P1, n and X are as defined above) is stirred to give a derivative bicyclic of the general formula (8) (where P1, n and X are as define previously). P2 can be removed by known techniques. When P2 is t-butyl group, it can be promptly removed using trifluoroacetic acid or a solution of hydrogen chloride / dioxane. When P2 is benzyl group, it can be readily removed using palladium carbon in combination with hydrogen or ammonium formate. When P2 is tetrahydropyranyl group, it can be promptly removed using acetic acid, p-toluenesulfonic acid or hydrochloric acid.

Step 4 (Process B) In this step, the bicyclic derivative of the general formula (8) (wherein P1, n and X are as defined above) is esterified or alkylated to obtain a bicycloester derivative of the general formula (9) (where R1, P1, n and X are as defined above). When the bicycloester derivative of the general formula (9) (wherein R1, P1, n and X are as defined above), the bicyclo derivative of the general formula (8) is produced by esterification (wherein P1, n and X are as defined above) is esterified with an alcohol derivative represented by R x OH (wherein R 1 is as defined above) in the presence of a condensing agent to give the bicycloester derivative of the general formula (9) wherein R 1 , P1, n and X are like define previously). Examples of condensation agents for esterification in this step include dicyclohexylcarbodiimide (DCC), 3-ethyl-1- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI), dimethylimidazolinium chloride (DMC), ethyl chloroformate, isobutyl chloroformate and pivaloyl chloride. These agents can be added in the form of solid, liquid or a solution in an appropriate solvent. When it is desired to use a base in the condensation reaction, the base may be an alkaline carbonate, such as sodium bicarbonate and potassium carbonate or a tertiary amine, such as triethylamine, diisopropylethylamine, N-methylmorpholine, diazabicyclo [5.4.0] -7-undecene, pyridine, 4-dimethylaminopyridine and 1,8-bis (dimethylamino) naphthalene. The solvent for use in the condensation reaction may be an inert solvent such as N, N-dimethylformamide, N, N-dimethylacetamide, dimethyl sulfoxide, acetonitrile, tetrahydrofuran, dioxane, ethyl ether, dimethoxyethane, ethyl acetate, toluene and dichloromethane. This condensation reaction proceeds smoothly at -20 to 150 ° C. When the bicycloester derivative of the general formula (9) (wherein R1, P1, n and X are as defined above) is produced by alkylation, the bicyclic derivative of the general formula (8) (wherein P1, n and X are as defined above) is reacted with a compound represented by RY (wherein Y is Cl, Br, I, OMs, OTs or OTf, and R1 is as defined above) in the presence in the absence of a base to give the bicycloester derivative of the general formula (9) (where R1, P1, n and X are as defined above). When it is desired to use a base in the reaction, the base may be an alkaline carbonate, such as sodium bicarbonate, potassium carbonate and cesium carbonate or a tertiary amine, such as triethylamine, diisopropylethylamine, N-methylmorpholine, diazabicyclo [5.4. 0] -7-undecene, pyridine, 4-dimethylaminopyridine and 1,8-bis (dimethylamino) naphthalene and phosphazene base and pentaisopropylguanidine. When it is desired to use a catalyst in the reaction, the catalyst can be a transfer phase catalyst or inorganic salt, such as tetrabutylammonium bromide, tetrabutylammonium iodide, benzyltriethylammonium bromide, lithium bromide, lithium iodide, sodium iodide, potassium bromide, potassium iodide, cesium bromide and cesium iodide. The solvent for use in the reaction may be an inert solvent such as acetone, ethanol, N, N-dimethylformate, N, N-dimethylacetamide, dimethyl sulfoxide, acetonitrile, tetrahydrofuran, dioxane, ethyl ether, dimethoxyethane, ethyl acetate, toluene and dichloromethane. This reaction proceeds smoothly at -30 to 150 ° C.

Step 5 (Process B) In this step, the bicyclic derivative of the general formula (8) (wherein P1, n and X are as defined above) is converted to a bicycloester derivative of the general formula (10) [wherein W is a reaction residue (such as halogen atoms, and halides, imidazolides, and carboxylic acid active esters, such as 1-imidazolyl group, 4-nitrophenoxy group, pentafluorophenoxy group, imidoyloxy succinate group, and 1-benzotriazolyl group ( or 3-benzotriazolyl group 3-oxide, P1, n and X are as described above.] This step can be easily carried out by known techniques When W is imidoyloxy succinate group, the bicyclic derivative of the general formula ( 8) (where P1, n and X are as defined above) is reacted with N-hydroxysuccinic acid in the presence of a condensing agent to give the desired product When W is benzotriazolyl group (or 3-oxide group of 1) -benzotriazolyl), the bicyclo derivative of the general formula (8) (wherein P1, n and X are as defined above) is reacted with 1-hydroxybenzotriazole in the presence of a condensing agent to give the desired product. Examples of the condensing agent for use in this step include dicyclohexylcarbodiimide (DCC), 3-ethyl-1- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI), dimethylimidazolinium chloride (DMC), ethyl chloroformate, isobutyl chloroformate and pivaloyl chloride. These agents can be added in the form of solid, liquid or a solution in an appropriate solvent. When it is desired to use a base in the condensation reaction, the base may be an alkaline carbonate, such as sodium bicarbonate or potassium carbonate, or a tertiary amine, such as triethylamine, diisopropylethylamine, N-methylmorpholine, diazabicyclo [5.4.0 ] -7-undecene, pyridine, 4-dimethylaminopyridine and 1,8-bis (dimethylamino) naphthalene. The solvent for use in the condensation reaction may be an inert solvent such as N, -dimethylformamide, N, N-dimethylacetamide, dimethyl sulfoxide, acetonitrile, tetrahydrofuran, dioxane, ethyl ether, dimethoxyethane, ethyl acetate, toluene and dichloromethane. This condensation reaction proceeds smoothly at -20 to 150 ° C. The resulting bicyclic derivative of the general formula (1) (wherein W, P1, n and X are as described above) can be used in the subsequent step after carrying out isolation and purification or as the raw product without purification.

Stage 6 (Process B) In this stage, the bicyclic derivative of the general formula (10 (wherein W, P1, n and X are as described above) is reacted with alcohol derivative represented by R1OH (wherein R1 is as defined above) to give a bicycloester derivative of the general formula ( 9) (where R1, P1, n and X are as described above). When a base is used in the reaction, the base may be an inorganic salt, such as sodium hydroxide, potassium hydroxide, sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate and cesium carbonate, or a base organic, such as triethylamine, diisopropylethylamine, N, N, N, N-tetramethylenediamine, diazabicyclo [5.4.0] -7-undecene, diazabicyclo [4.3.0] -nonne, 4-dimethylaminopyridine, phosphazene base and pentaisopropylguanidine. The solvent for use in the reaction may be an inert solvent such as toluene, acetonitrile, tetrahydrofuran, dioxane, ethyl ether, t-butyl methyl ether, dimethoxyethane, ethyl acetate, dichloromethane, N, N-dimethylformamide, dimethylsulfoxide and N-methyl-2. pyrrolidine. This reaction proceeds smoothly at -30 to 150 ° C.

Step 7 (Process B) In this step, the group P1 protecting the secondary amino group in the bicycloester derivative of the general formula (9) (wherein Ra, P1, n and X are as defined above) is removed to give a bicycloester derivative of the general formula (1) (wherein R1, n and X are as defined above). P1 can be removed by known techniques. When P1 is t-butoxycarbonyl group, it can be promptly removed using trifluoroacetic acid or a solution of hydrogen chloride / dioxane. When P1 is benzyloxycarbonyl group, it can be readily removed using palladium carbon in combination with hydrogen or ammonium formate. When P1 is trifluoroacetyl group, it can be promptly removed using an ammonium / methanol solution.

Process C Step 1 (Process C) In this step, a bicycloester derivative of the general formula (1) (wherein R1, n and X are as defined above) is obtained through esterification or alkylation. When the bicycloester derivative of the general formula (1) (wherein R1, n, and X are as defined above) is obtained through esterification, a bicyclic derivative of the general formula (11) (wherein n and X are as defined above) is esterified with an alcohol derivative represented by R 1 OH (wherein R 1 is as defined above) in the presence of a condensing agent to achieve the bicycloester derivative of the general formula (1) (wherein R1, n and X are as defined above). Examples of the condensing agent for use in the esterification include dicyclohexylcarbodiimide (DCC), 3-ethyl-1- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI), dimethylimidazolinium chloride (DMC), ethyl chloroformate, isobutyl chloroformate and chloride of pivaloyl. These agents can be added in the form of solid, liquid or a solution in an appropriate solvent. When it is desired to use a base in the condensation reaction, the base may be an alkaline carbonate, such as sodium bicarbonate and potassium carbonate, or a tertiary amine, such as triethylamine, diisopropylethylamine, N-methylmorpholine, diazabicyclo [5.4.0 ] -7-undecene, pyridine, 4-methylaminopyridine and 1,8-bis (dimethylamino) naphthalene. The solvent for use in the condensation reaction may be an inert solvent such as N, -dimethylformamide, N, N-dimethylacetamide, di-ethyl sulfoxide, acetonitrile, tetrahydrofuran, dioxane, ethyl ether, dimethoxyethane, ethyl acetate, toluene and dichloromethane. This condensation reaction proceeds smoothly at -20 to 150 ° C. Alternatively, the condensation reaction can be carried out via an active ester or acid chloride having 1-imidazolyl group, 4-nitrophenoxy group, pentafluorophenoxy group, imidoyloxy succinate group or 1-benzotriazolyloxy group (or group 3- 1-benzotriazolyl oxide). In such a case, the active ester or acid chloride can be used in the subsequent step after carrying out isolation and purification or as the raw product without purification. When the bicycloester derivative of the general formula (1) (wherein R1, n and X are as defined above), a bicyclic derivative of the general formula (11) is obtained by alkylation (where n and X are as defined above) is reacted with a compound represented by R ^? 2 (wherein Y2 and R1 are as described above) in the presence or absence of a base to give the bicycloester derivative of the general formula (1) (in where R1, n and X are as defined above). When it is desired to use a base in the reaction, the base may be an alkaline carbonate, such as sodium bicarbonate, potassium carbonate and cesium carbonate or a tertiary amine, such as triethylamine, diisopropylethylamine, N-methylmorpholine, diazabicyclo [5.4.0] -7-undecene, pyridine, 4-dimethylaminopyridine and 1,8-bis (dimethylamino) naphthalene and phosphazene base and pentaisopropylguanidine. When you want to use a catalyst in the reaction, the catalyst can be a transfer phase catalyst or an inorganic salt, such as tetrabutylammonium bromide, tetrabutylammonium iodide, benzyltriethylammonium bromide, lithium bromide, lithium iodide, sodium iodide, potassium bromide, potassium iodide, cesium bromide and cesium iodide. The solvent for use in the reaction may be an inert solvent such as acetone, ethanol, N, N-dimethylformamide, N, N-dimethylacetamide, dimethyl sulfoxide, acetonitrile, tetrahydrofuran, dioxane, ethyl ether, dimethoxyethane, ethyl acetate, toluene and dichloromethane. This reaction proceeds smoothly at -30 to 150 ° C. The advantageous features of the present invention will now be described with reference to experiments and examples, which are not intended to limit the scope of the invention in any way.

Reference Example 1 Synthesis of ethyl 4-aminobicyclo [2.2.2] octane-1-carboxylate Step 1: Synthesis of methyl 4-benzyloxycarbonylamino-bicyclo [2.2.2] octane-1-carboxylate Bicyclo [2.2.2] octane-1,4-dicarboxylate methyl hydrogen (25.0 g), diphenylphosphoryl azide ( 32.5 g), triethylamine (17.3 ml) and toluene (500 ml) were mixed together. The mixture was stirred for 2 hours at room temperature and heated to reflux for 2 hours. To the resulting mixture, benzyl alcohol was added (122 ml) and the mixture was further heated and refluxed for 17 hours. Subsequently, the mixture was allowed to cool and was subsequently washed with a 10% aqueous citric acid, saturated aqueous sodium bicarbonate solution and saturated brine. The mixture was then dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: hexane: ethyl acetate = 2: 1) to give methyl 4-benzyloxycarbonylaminobicyclo [2.2.2] octane-1-carboxylate (32.2 g). MS (FAB +) m / z. 318 (MH +).

Step 2: Synthesis of 4-benzyloxycarbonylaminobicyclo [2.2.2] octane-1-carboxylic acid. [0129] 4-Benzyloxycarbonylaminobicyclo [2.2.2] octane-1-carboxylic acid methyl ester (64.3 g) was dissolved in ethanol (1100 ml). For this solution, a 1 mol / 1 aqueous solution of sodium hydroxide was added and the mixture was stirred at 50 ° C for 1 hour. Ethanol was evaporated in the mixture under reduced pressure and the residue was washed with diethyl ether (500 ml), followed by the addition of concentrated hydrochloride acid to adjust the pH to acidic (pH 1). The resulting crystals were filtered, washed with water and dried under reduced pressure to give 4-benzyloxycarbonylaminobicyclo [2.2.2] octane-1-carboxylic acid (56.1 g). MS (FAB +) m / z: 304 (MH +).

Step 3: Synthesis of ethyl 4-benzyloxycarbonylamino-bicyclo [2.2.2] octane-1-carboxylate. 4-Benzyloxycarbonylaminobicyclo [2.2.2] octane acid (56.0 g) was dissolved in N, N-dimethylformamide (1000 ml). To this solution, sodium bicarbonate (46.6 g) and then ethyl iodide (22.2 ml) were added and the mixture was stirred at 50 to 60 ° C for 5 hours.

Subsequently, additional sodium bicarbonate (46.6 g) and ethyl iodide (22.2 ml) were added and the mixture was stirred for an additional 3 hours. Insoluble materials in the mixture were filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (700 ml) and the solution was washed with water, dried over anhydrous sodium sulfate and dried under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: hexane: ethyl acetate = 2: 1 → ethyl acetate) to give ethyl 4-benzyloxycarbonylamino-bicyclo [2.2.2] -octane-1-carboxylate (56.8 g). MS (FAB +) m / z: 332 (MH +).

Step 4: Synthesis of ethyl 4-aminobicyclo [2.2.2] octane-1-carboxylate Ethyl 4-benzyloxycarbonylaminobicyclo [2.2.2] octane-1-carboxylate (40.0 g) was dissolved in ethanol (40 ml). To this solution, palladium on 10% carbon (4.00 g) was added and the mixture was stirred at room temperature for 6 hours in a stream of hydrogen. The catalyst in the reaction mixture was filtered through a pad of celite and the catalyst was filtered, along with the pad of Celite, it was washed with ethanol. The filtrate and the washings were combined and concentrated under reduced pressure. The resulting residue was dried under reduced pressure to give ethyl 4-aminobicyclo [2.2.2] octane-1-carboxylate (23.9 g). MS (EI +) m / z: 197 (M +).

Reference Example 2 Synthesis of 1,1-dimethylethyl 4-aminobicyclo [2.2.2] octane-1-carboxylate Step 1: Synthesis of 1,1-dimethylethylmethyl bicyclo [2.2.2] octane-1,4-dicarboxylate dissolved bicyclo [2.2.2] octane-1,4-dicarboxylic acid methyl ester (500 mg) in dichloromethane (5 ml) and sulfuric acid (50 μl) was added to the solution. The mixture was bubbled with isobutene for 5 minutes while cooling in a salt / ice bath. The mixture was then stirred for 4 hours at room temperature and left for 4 days. Subsequently, the reaction mixture was diluted with dichloromethane (5 ml), washed sequentially with a saturated aqueous solution of sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was dried under reduced pressure to give 1,1-dimethylethylmethyl bicyclo [2.2.2] octane-1,4-dicarboxylate (497 mg).

MS (FAB +) m / z: 269 (MH +).

Step 2: Synthesis of 1, 1-dimethylethyl hydrogen [2.2.2] octan-2-1, 4-dicarboxylate hydrogen Bicyclo [2.2.2] octane-1,4-dicarboxylic acid 1,1-dimethylethyl methyl ester was dissolved ( 495 mg) in methanol (5 ml). To this solution, an aqueous solution of 2 mol / l sodium hydroxide (0.92 ml) was added and the mixture was stirred at room temperature for 8 hours and then at 50 ° C for 2 hours. The methanol in the mixture was evaporated under reduced pressure. Water was added to the resulting residue and the mixture was washed with diethyl ether and neutralized with 3 mol / l hydrochloric acid. The resulting crystals were filtered, washed with water and dried under reduced pressure to give 1,1-dimethylethyl hydrogen bicyclo [2.2.2] octane-1,4-dicarboxylate (344 mg).

Step 3: Synthesis of 1,1-dimethylethyl 4-benzyloxycarbonylamino-bicyclo [2.2.2] octane-1-carboxylate Using bicyclo [2.2.2] octane-1,4-dicarboxylate 1,1-dimethylethyl hydrogen (340 mg ), the same procedure was followed as in Step 1 of Reference Example 1 to give 4-benzyloxycarbonylaminobicyclo [2.2.2] octan-1- 1,1-dimethylethyl carboxylate (433 mg). XH NMR (CDC13) d 1.41 (s, 9H), 1.84 (s, 12H), 4.48-4.62 (br, ÍH), 5.03 (s, 2H), 7.28-7.38 (m, 5H).

Stage 4: Synthesis of 1,1-dimethylethyl 4-aminobicyclo [2.2.2] octane-1-carboxylate Using 1,1-dimethylethyl-4-benzyloxycarbonylamino bicyclo [2.2.2] octane-1-carboxylate (425 mg) , the same procedure was followed as in Step 4 of the Example of Reference 1 to give 4-aminobicyclo [2.2.2] octane-1-carboxylate 1,1-dimethylethyl (218 mg). XH NMR (CDC13) d 1.41 (s, 9H), 1.52-1.56 (m, 6H), 1.80-1.84 (m, 6H).

Reference Example 3 Synthesis of bicyclo [2.2.2] octane-1-carboxylate of 2-tetrahydropyranol 4-benzyloxycarbonylamino 4-benzyloxycarbonylaminobicyclo [2.2.2] -octane-1-carboxylic acid was suspended (1.00 g) in dichloromethane (10 ml). To this suspension, 3,4-dihydro-2H-pyran (1.20 ml) and then p-toluenesulfonic acid monohydrate (6.3 mg) were added and the mixture was stirred at room temperature for 30 minutes. Subsequently, the reaction mixture was washed sequentially with a solution aqueous saturated sodium bicarbonate and water, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: hexane: ethyl acetate = 4: 1) to give bicyclo [2.2.2] octane-1-carboxylate of 2-tetrahydropyranyl 4-benzyloxycarbonylamino (1.18 g). 2H NMR (CDC13) d 1.53-1.95 (m, 18H), 3.67-3.71 (m, ÍH), 3.82-3.89 (m, ÍH), 4.59 (br, ÍH), 5.03 (s, 2H), 5.95 (br , ÍH), 7.29-7.38 (, 5H).

Step 2: Synthesis of 2-tetrahydropyranyl 4-aminobicyclo [2.2.2] octane-1-carboxylate Using 4-benzyloxycarbonylamino-bicyclo [2.2.2] octane-1-carboxylate of 2-tetrahydropyranyl (548 mg), the same procedure as in Step 4 of Reference Example 1 to give 4-aminobicyclo [2.2.2] octan-1-carboxylate of 2-tetrahydropyranyl (357 mg). MS (EI +) m / z: 253 (M +).

Reference Example 4 Synthesis of (2S, 4S) -1- (2-chloroacetyl) -4-fluoropyrrolidine-2-carbonitrile In accordance with the production process of (2S, 4S) -1- (2-bromoacetyl) -4-fluoropyrrolidine-2-carbonitrile described in WO 02/38541, (2S, 4S) -4-fluoropyrrolidine-2-carboxamide hydrochloride (5.00 g) was used. ) and chloroacetyl chloride (2.60 ml) to obtain (2S, 4S) -1- (2-chloroacetyl) -4-fluoropyrrolidine-2-carbonitrile (4.96 g). MS (EI +) m / z: 190 (M +). HRMS (EI +) for C7H8C1FN20 (M +): caled, 190.0309; found, 190.0283.

Example 1 Synthesis of (2S, 4S) -l- [[N- (4-ethoxycarbonyl bicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile 4-aminobicyclo [2.2.2] Ethyl octane-1-carboxylate (92.0 mg) was dissolved in acetonitrile (2 ml) and diisopropylamine (100 μl) was added to the solution. While the mixture was cooled in an ice bath, it was added dropwise in acetonitrile (1 ml) (2S, 4S) -1- (2-bromoacetyl) -4-fluoropyrrolidine-2-carbonitrile (100 mg). The mixture was stirred for 1.5 hours while cooling in an ice bath. Subsequently, water was added and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed with Saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: ethyl acetate: methanol = 5: 1) to give (2S, 4S) -l- [[N- (4-ethoxycarbonylbicyclo [2.2.2] oct- 1-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (132 mg). MS (EI +) for C? 8H26FN303 (M +): caled, 351. 1958; found, 351 1982 Example 2 Synthesis of (2S) -l- [[N- (4-ethoxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] pyrrolidine-2-carbonitrile 4-aminobicyclo [2.2.2] octane-1-carboxylate from ethyl (115 mg) was dissolved in N, N-dimethylformamide (1.8 ml) and diisopropylamine (100 μl) was added to the solution. Dr (2S) -1- (2-bromoacetyl) pyrrolidine-2-carbonitrile (120 mg) in N, -dimethylformamide (1 ml) was then added dropwise at room temperature. The mixture was stirred for 2 hours at room temperature and concentrated under reduced pressure. The residue was purified by gel column chromatography silica (eluent: chloroform: methanol = 10: 1) to give (2S) -1 - [[N- (4-ethoxycarbonylbicyclo [2.2.2] oct-1-yl) amino] acetyl] pyrrolidine-2-carbonitrile ( 95.1 mg). MS (EI +) m / z: 333 (M +). HRMS (EI +) for C? 8H27N303 (M +): caled, 333.2052; found, 333.2037.

Example 3 Synthesis of (2S) -l- [[N- (4-t-butoxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to Example 2, 4- was used. aminobicyclo [2.2.2] octane-1-carboxylic acid t-butyl ester (100 mg) and (2S) -1- (2-bromoacetyl) pyrrolidine-2-carbonitrile (90.0 mg), to obtain (2S) -1- [ [N- (4-t-Butoxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] pyrrolidine-2-carbonitrile (97.6 mg). MS (EI +) m / z: 361 (M +). HRMS (EI +) for C20H3? N3O3 (M +): caled, 361.2365; found, 361.2373.

Example 4 Synthesis of (2S, 4S) -l- [[N- [4- (2-tetrahydropyranyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile In a similar way to Example 2, 4-aminobicyclo [2.2.2] octane-1-carboxylate of 2-tetrahydropyranyl (62.9 mg) and (2S, 4S) -1- (2-bromoacetyl) -4-fluoropyrrolidine-2-carbonitrile ( 53.1 mg) to obtain (2S, 4S) -l- [[N- [4- (2-tetrahydropyranyl) oxycarbonylbicyclo [2.2.2] oct-1-yl] amino] acetyl] 4-fluoropyrrolidine-2-carbonitrile (73.3 mg). MS (FAB +) m / z: 408 (M +). HRMS (FAB +) for C2? H3aFN304 (MH +): caled, 408. 2299; found, 408 2295 Example 5 Synthesis of (2S9-1 - [[N- [4- (2-tetrahydropyranyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to Example 2, it was used 4-Aminobicyclo [2.2.2] octan-1-carboxylate of 2-tetrahydropyranyl (90.0 mg) and (2S9-l- (2- bromoacetyl) pyrrolidine-2-carbonitrile (70.0 mg), to obtain (2S) -1- [[N- [4- (2-tetrahydropyranyl) oxycarbonyl bicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine -2-carbonitrile (85.2 mg). MS (EI +) m / z: 389 (M +). HRMS (EI +) for C2? H3? N304 (M +): caled, 389. 2315; found, 389 2296 E j us 6 Synthesis of (2S, 4S) -l- [[N- (4-ethoxycarbonylbicyclo [2.2.1] hept-1-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile 4-aminobicyclo [2.2.1] was dissolved ] ethyl heptan-1-carboxylate (50.0 mg) in N, N-dimethylformamide (2 ml) and potassium carbonate (40.0 mg) was added to the solution. Then (2S, 4S) -1- (2-bromoacetyl) -4-fluoropyrrolidine-2-carbonitrile (64.2 mg) in N, N-dimethylformamide (1 ml) was added dropwise at room temperature and the mixture was stirred for 1 hour. hour. The insoluble materials were filtered and the filtrate was concentrated. The resulting residue was purified by column chromatography on silica gel (NH silica, eluent: ethyl acetate) to give (2S, 4S) -1- [[N- (4-ethoxycarbonylbicyclo [2.2.1] hept-1-yl) amino] acetyl] -4-fluoropyrrolidin-2-carbonitrile (89.0 mg). MS (FAB +) m / z: 338 (M +). HRMS (FAB +) for C? 7H25FN303 (MH +): caled, 338.1880; found, 338.1835.

Example 7 Synthesis of (2S) -1- [[N- (4-ethoxycarbonylbicyclo [2.2.1] hept-1-yl) amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to Example 6, 4-aminobicyclo [ 2.2.1] ethyl eptan-1-carboxylate (50.0 mg) and (2S) -1- (2-bromoacetyl) pyrrolidine-2-carbonitrile (59.3 mg), to obtain (2S) -1- [[N- ( 4-Ethoxycarbonylbicyclo [2.2.1] hept-1-yl) amino] acetyl] pyrrolidine-2-carbonitrile (79.4 mg). MS (FAB +) m / z: 320 (M +). HRMS (FAB +) for C? 7H26N303 (MH +): caled, 320. 1974; found, 320 1975.

Example 8 Synthesis of (2S, 4S) -l- [[N- (4-benzyloxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile It was dissolved (2S, 4S) -l - [[N- (4-carboxybicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (30.0 mg) in N, N-dimethylformamide (1.0 ml) and cesium carbonate (45.3 mg) was added to the solution. Benzyl bromide was then added (17.5 mg) in N, N-dimethylformamide (0.5 ml), while the mixture was cooled in an ice bath and the mixture was stirred for 1 hour. Subsequently, water was added and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel (eluent: dichloromethane: methanol = 10: 1) to give (2S, 4S) -1- [[N- (4-benzyloxycarbonyl bicyclo [2.2.2] oct- 1-yl) amino] acetyl-4-fluoropyrrolidine-2-carbonitrile (30.6 mg). MS (FAB +) m / z: 414 (M +). HRMS (FAB +) for C25H29FN303 (MH +): caled, 414.2193; found, 414.2176 Example 9 Synthesis of (2S, 4S) -1- [[N- (4-cyclopropylmethyloxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile In a manner similar to Example 8, use (2S, 4S) -l- [[N- (4-carboxybicyclo [2.2.2] oct-l-yl) amino] acetyl-4-fluoropyrrolidine-2-carbonitrile (20.0 mg) and cyclopropylmethyl bromide (12.8 mg) , to obtain (2S, 4S) -l - [[N- (4-cyclopropylmethyloxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (13.7 mg). MS (FAB +) m / z: 378 (M +). HRMS (FAB +) for C20H29FN3O3 (MH +): caled, 378.2193; found, 378.2207.

Example 10 Synthesis of (2S, 4S) -4-fluoro-1- [[N- [4- (4-trifluoromethylbenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to the example, 8, (2S, 4S) -l- [[N- (4-carboxybicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (20.0 mg ) and 4-trifluoromethylbenzyl bromide (16.2 mg), to obtain (2S, 4S) -4-fluoro-1- [[N- [4- (4-trifluoromethylbenzyl) oxycarbonylbicyclo [2.2.2] oct-1-yl] amino] acetyl] pyrrolidine-2-carbonitrile (22.2 mg). MS (FAB +) m / z: 482 (M +). HRMS (FAB +) for C2H28F4N303 (MH +): caled, 482.2067; found, 482.2068.

Example 11 Synthesis of (2S, 4S) -4-fluoro-1- [[N- [4- (2-methylpropyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to Example 8, (2S, 4S) -1- [[N- (4-carboxybicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (20.0 mg) was used and isobutyl bromide (9.3 mg), to obtain (2S, 4S) - 4-Fluoro-l- [[N- [4- (2-methylpropyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile (14.2 mg). MS (FAB +) m / z: 380 (M +). HRMS (FAB +) for C20H31FN3O3 (MH +): caled, 380.2349; found, 380.2361.

Example 12 Synthesis of (2S, 4S) -4-fluoro-l- [[N- [4- (2-methylethyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to Example 8, was used (2S, 4S) -1- [[N- (4-carboxybicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidin-2-carbonitrile (20.0 mg) and isopropyl iodide (10.5 mg ), to obtain (2S, 4S) -4-fluoro-l- [[N- [4- (2-methylethyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile ( 11.2 mg). MS (EI +) m / z: 365 (M +). HRMS (EI +) for C? 9H28FN303 (M +): caled, 365.2115; found, 365.2096.

Example 13 Synthesis of (2S, 4S) -l- [[N- [4- (4-ethoxycarbonylbenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile In a similar manner to Example 8, (2S, 4S) -1- [[N- (4-carboxybicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (32.3 mg) was used and 4-Ethoxycarbonylbenzyl bromide (28.1 mg), to obtain (2S, 4S) -1- [[N- [4- (4-ethoxycarbonylbenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] -4 -fluoropyrrolidin-2-carbonitrile (34.7 mg). MS (EI +) m / z: 485 (M +). HRMS (EI +) for C26H32FN305 (M +): caled, 485.2326; found, 485.2309.Example 14 Synthesis of (2S, 4S) -l- [[4- [4- (2,2-dimethylethyl) benzyl] oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl-4-fluoropyrrolidine-2-carbonitrile In a similar manner to Example 8, was used (2S, 4S) -1- [[N- (4-carboxybicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (30.0 mg) and 4- (2-bromide , 2-dimethylethyl) benzyl (23.2 mg) to obtain (2S-4S) -1- [[N- [4- [4- (2, 2-dimethylethyl) benzyl] oxycarbonylbicyclo [2.2.2] oct-l-yl ] amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (29.9 mg). MS (FAB +) m / z: 470 (M +). HRMS (FAB +) for C27H37FN303 (MH +): caled, 470.2819; found, 470.2859.

Example 15 Synthesis of (2S, 4S) -l- [[N- [4- (4-chlorobenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl-4-fluoropyrrolidin-2-carbonitrile In a manner similar to Example 8, (2S, 4S) -l- [[N- (4-carboxybicyclo [2.2.2] oct-1-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (30.0 mg) and bromide was used of 4-chlorobenzyl (21.0 g), to obtain (2S, 4S) -l - [[N- [4- (4-chlorobenzyl) oxycarbonylbicyclo [2.2.2] oct-1-yl] amino] acetyl] -4- fluoropyrrolidin-2-carbonitrile (25.5 mg). MS (FAB +) m / z: 448 (M +). HRMS (FAB +) for C23H28FN303 (MH +): caled, 448.1803; found, 448.1794.

Example 16 Synthesis of (2S, 4S) -4-fluoro-l- [[N- [4- (4-methylbenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to Example 8, (2S, 4S) -1- [[N- (4-carboxybicyclo [2.2.2] oct-1-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (30.0 mg) was used and 4-methylbenzyl bromide (18.9 mg), to obtain (2S, 4S) -4-fluoro-l- [[N- [4- (4-methylbenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino ] acetyl] pyrrolidine-2-carbonitrile (9.2 mg). MS (FAB +) m / z: 428 (M +). HRMS (FAB +) for C 24 H 31 FN 303 (MH +): caled, 428.2349; found, 428.2382.

Example 17 Synthesis of (2S, 4S) -4-fluoro-l- [[N- [4- (4-methoxybenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to Example 8, (2S, 4S) -1- [[N- (4-carboxybicyclo [2.2.2] oct-1-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (30.0 mg) was used and 4-methoxybenzyl chloride (16.0 mg), to obtain (2S, 4S) -4-fluoro-l- [[N- [4- (4-methoxybenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino ] acetyl] pyrrolidine-2-carbonitrile (29.8 mg). MS (FAB +) m / z: 444 (M +). HRMS (FAB +) for C 24 H 31 FN 303 (MH +): caled, 444.2299; found, 444.2269.

Example 18 Synthesis of (2S, 4S) -4-fluoro-l- [[N- [4- (2-trifluoromethylbenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to Example 8, (2S, 4S) -l- [[N- (4-carboxybicyclo [2.2.2] oct-l- was used il) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (30.0 g) and 2,6-dichlorobenzyl bromide (24.5 mg), to obtain (2S, 4S) -4-fluoro-l- [[N- [ 4- (2-trifluoromethylbenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile (39.9 mg). MS (EI +) m / z: 481 (M +). HRMS (EI +) for C 24 H 27 F 4 N 303 (M +): caled, 481.1989; found, 481.1944.

Example 19 Synthesis of (2S, 4S) -1- [[N- [4- (2,6-dichlorobenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile In a Similar to Example 8, (2S, 4S) -l- [[N- (4-carboxybicyclo [2.2.2] oct-1-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (30.0 mg ) and 2,6-dichlorobenzyl bromide (24.5 mg), to obtain (2S, 4S) -l - [[N- [4- (2,6-dichlorobenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] -4-fluoropyrrolidin-2-carbonitrile (40.0 mg). MS (EI +) m / z: 481 (M +).

HRMS (EI +) for C23H26C12FN303 (M +): caled, 481.1335; found, 481.1366.

Example 20 Synthesis of (2S, 4S) -4-fluoro-l- [[N- [4- (2, 3, 4, 5, 6-pentafluorobenzyl) oxycarbonylbicyclo [2.2.2] oct-1-yl] amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to Example 8, (2S, 4S) -1- [[N- (4-carboxycyclo [2.2.2] oct-1-yl) amino] acetyl] -4-fluoropyrrolidine was used. -2-carbonitrile (30.0 mg) and 2, 3, 4, 5, 6-pentafluorobenzyl bromide (26.6 mg), to obtain (2S, 4S) -4-fluoro-l- [[N- [4- (2 , 3, 4, 5, 6-pentaf luorobenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile (41.9 mg). MS (EI +) m / z: 503 (M +). HRMS (EI +) for C23H23F6N303 (M +): caled, 503.1644; found, 503.1681.

Example 21 Synthesis of (2S, 4S) -4-fluoro-1- [[N- [4- (2-methylbenzyl) oxycarbonylbicyclo [2.2.2] oct-l-yl] amino] acetyl] pyrrolidine-2-carbonitrile In a manner similar to Example 8, (2S, 4S) -l- [[N- (4-carboxybicyclo [2.2.2 [oct-l-yl] amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (30.0 mg) was used and 2-methylbenzyl bromide (18.9 mg), to obtain (2S, 4S) -4-fluoro-l- [[N- [4- (2-methylbenzyl) oxycarbonylbicyclo [2.2.2] oct-1-yl] amino] ] acetyl] pyrrolidine-2-carbonitrile (34.1 mg). MS (EI +) m / z: 427 (M +). HRMS (EI +) for C24H30FN3O3 (M +): caled, 427.2271; found, 427.2312.

Example 22 Synthesis of (2S, 4S, 1 'S) -1- [[N- (1-phenylethyloxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile They were dissolved (2S, 4S) -1- [[N- (4-carboxybicyclo [2.2.2] oct-l-yl] amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (50.0 mg), 1-hydroxybenzotriazole (26.0 mg), hydrochloride of l- (3- dimethylaminopropyl) -3-ethylcarbodiimide (32.5 mg), and (lS) -phenethyl alcohol (0.094 ml), in N, -dimethylformamide (0.9 ml). The mixture was stirred at room temperature for 22 days and subsequently concentrated under reduced pressure. The resulting residue was purified by thin layer chromatography (eluent: dichloromethane: methanol = 10: 1) to give (2S, S, 1 'S) -1- [[N- (1-phenethyloxycarbonylbicyclo [2.2.2] oct- 1-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (4.6 mg). MS (FAB +) m / z: 428 (M +). HRMS (FAB +) for C2H3? FN303 (MH +): caled, 428.2349; found, 428.2369.

Example 23 Synthesis of (2S, 4S, 1 'R) -1- [[N- (l-phenylethyloxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile In a manner similar to Example 22, was used (2S, 4S) -l- [[N- (4-carboxybicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (50.0 mg), 1-hydroxybenzotriazole (26.0 mg) ) and (1R) -phenethyl alcohol (0.094 ml), to obtain (2S, 4S, 1 'R) -1- [[N- (1- phenylethyloxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (3.6 mg). MS (FAB +) m / z: 428 (M +). HRMS (FAB +) for C24H3? FN303 (MH +): caled, 428. 2349; found, 428.2342.

Example 24 Synthesis of (2S, 4S) -4-fluoro-l- [[N- (5-hydroxypentyloxycarbonylbicyclo [2.2.2] oct-l-yl) amino] acetyl] pyrrolidine-2-carbonitrile They were mixed together (2S, 4S ) -1- [[N- (4-carboxybicyclo [2.2.2] oct-l-yl) amino] acetyl] -4-fluoropyrrolidine-2-carbonitrile (50.0 mg, 5-bromo-l-pentanol (18.7 μl) , potassium carbonate (23.5 mg), and N, N-dimethylformamide (1.5 ml) The mixture was stirred at room temperature for 4 hours.The insoluble materials in the mixture were filtered and the filtrate was concentrated under reduced pressure. The resulting product was purified by a silica gel column (eluent: chloroform: methanol = 10: 1) to give (2S, 4S) -4-fluoro-l- [[N- (5-idroxypentyloxycarbonylbicyclo [2.2.2] oct- 1-yl) amino] acetyl] pyrrolidine-2-carbonitrile (32.4 mg).

MS (FAB +) m / z: 410 (M +). HRMS (FAB +) for C2? H33FN304 (MH +): caled, 410. 2455; found, 410.2420.

Test Example 1 Test to determine the ability of the compounds of the invention to inhibit the activity of dipeptidylpeptidase IV The concentration of free 7-amino-4-methyl-coumarin (AMC) generated by hydrolysis of H-Gly-Pro-AMC « HBr subtracted by dipeptidylpeptidase IV from the plasma was determined by fluorometry.

Method 20 μl of buffer (25 mmol / l hepes, 140 mmol / l of sodium chloride, 1% bovine serum albumin, 80 mmol / l of magnesium chloride hexahydrate, pH 7.4) containing each compound, was added to 20 μl of 8 parts diluted plasma with saline in a cavity of a 96-well flat bottom plate. The plate was left at room temperature for 5 minutes and 10 μl of 0.1 mmol / l of H-Gly-Pro-AMC "HBr solution was added to each well to initiate the reaction. The plate was left in a dark environment at room temperature for 20 minutes, at which point, 20 μl of 25% acetic acid was added to finish the reaction. Using a fluorescent plate reader, the concentration of free AMC was determined by exciting the samples at 355 nm and measuring the fluorescence intensity at 460 nm. Using Prima 3.02 (GraphPad Software), the results were analyzed to determine 50% inhibitory concentration (IC50). The results are shown in Table 1.

Table 1 Inhibitory activity of dipeptidylpeptidase IV in vitro Compound A: (2S) -1 - [[(3-hydroxy-l-adamantyl) amino] acetyl] -2-cyanopyrrolidine LAF-237) Test Example 2 Test to determine inhibition of dipeptidylpeptidase IV activity in mice by oral administration of the compounds of the invention Each compound was suspended in 0.3% sodium carboxymethylcellulose at a concentration of 0.1 mg / ml. The preparation was administered orally to 8-week old male ICR mice (Charles River Laboratories Japan) at a dose of 10 ml / kg. Using a capillary tube treated with EDTA 2K, blood samples were collected from the tail vein before administration and 30 minutes after administration. The blood samples were centrifuged at 6000 rpm for 2 minutes to separate the plasma. Enzyme activity was determined using the same procedure as in Test Example 1. Inhibition was determined from the decrease in enzyme activity from the initial activity (% inhibition =. {(Activity before administration -activity after of administration) / (activity before administration).}. x 100). Results are shown in table 2.

Table 2 Inhibition of dipeptidylpeptidase IV activity by oral administration Compound A: (2S) -1 - [[(3-hydroxy-l-adamantyl) amino] acetyl] -2-cyanopyrrolidine (LAF-237) Test Example 3 Oral glucose tolerance test in mice The compound of the present invention of Example 1 was suspended in 0.3% sodium carboxymethyl cellulose (CMC-Na, Sigma). Seven week old ICR male mice (Charles River Laboratories Japan), were acclimated for 1 week. During the acclimation period, the animals were allowed to freely consume standard food (CE-2, Clea Japan) and water. The ICR mice that reached 8 weeks of age were fasted for 16 hours. Subsequently, the animals were orally administered 0.2% CMC-Na (10 ml / kg) of Compound 1 (1 mg / kg, 10 ml / kg) and were immediately administered with a glucose solution orally at a dose of 5 g / kg. Using a capillary tube treated with EDTA 2K, blood samples were collected from the tail vein before administration of glucose solution and 15, 30, 60 and 120 minutes after administration. The level of glucose in the blood was determined using the Wako glucose test B (Wako Pure Chemical Industries). The results are shown in means + standard errors. Statistical analyzes were performed using the t test with a significant level of less than 5%. The results are shown in Figure 1.

Test Example 4 Test to determine the efficacy of the compounds of the invention against drug-induced hypoleukocytosis The efficacy of the compounds of the present invention against drug-induced hypoleukocytosis was evaluated by conducting an experiment in accordance with the method described by Okabe et al. ., (Japanese Pharmacology and Therapeutics, Vol. 19. No. 6 (1991): p55). Eight-week-old male ICR mice (Charles River Laboratories Japan), were administered intraperitoneally with a single dose of cyclophosphamide (200 mg / kg) at day 0. Starting the next day, the control group was given saline and the group of Test was orally administered with the compound of the present invention (1 to 200 mg / kg) once or twice a day for a period of five days. Blood samples were collected 2, 4, 6 and 8 days after the start of the test and the white blood cell count was monitored over time. The white blood cell count of the test group at a given time was compared to the white blood cell count before the administration of cyclophosphamide to evaluate the efficacy of the compound of the present invention against the drug-induced hypoleukocytosis. . The results indicate that the decrease in the counting of white blood cells was significantly suppressed in the group administered with the compound of the present invention, compared to the control group.

Test Example 5 Test to determine the ability of the compounds of the invention to increase the level of G-CSF in the blood Seven-week-old male ICR mice (Charles River Laboratories Japan) were used. The saline was given to the control group and the test group was administered orally with the compound of the present invention (1 to 200 mg / kg) once or twice a day for a period of five days. The mice were anesthetized the day after cessation of administration and blood samples were collected. The level of G-CSF in the plasma was determined using a mouse G-CSF ELISA kit (R &D SYSTEM). The results indicate that the plasma G-CSF level was significantly increased in the group administered with the compound of the present invention, compared to the control group.

INDUSTRIAL APPLICABILITY As discussed, the compounds of the present invention are novel bicycloester derivatives and salts of the same pharmaceutically acceptable having a high inhibitory activity of DPP-IV. The pharmaceutical compositions containing the compound of the present invention as an active ingredient, are useful in the prevention and / or treatment of diabetes and associated diabetic complications, as well as in the prevention and / or treatment of other diseases involving DPP-IV. .

Claims (6)

NOVELTY OF THE INVENTION Having described the present is considered as a novelty, and therefore, it is claimed as property contained in the following: CLAIMS

1. A bicycloester derivative, represented by the following general formula (1): characterized in that R1 is an alkyl group Ci to substituted or unsubstituted C,, substituted or unsubstituted C 3 to β-cycloalkyl group, tetrahydropyranyl group, substituted or unsubstituted arylmethyl group, substituted or unsubstituted arylethyl group, substituted or unsubstituted aromatic hydrocarbon group, substituted or unsubstituted aromatic heterocyclic ring, or substituted aliphatic heterocyclic ring or unsubstituted; X is CH2, CHF, CF2, CHOH, S, or O; n is 1, 2 or 3, or a pharmaceutically acceptable salt thereof.

2. An intermediary in the production of the derivative of bicycloester according to claim 1, represented by the following general formula (2): (2) characterized in that R1 is a substituted or unsubstituted Ci to C6 alkyl group, substituted or unsubstituted C3 to C6 cycloalkyl group, tetrahydropyranyl group, substituted or unsubstituted arylmethyl group, substituted or unsubstituted arylethyl group, substituted or unsubstituted aromatic hydrocarbon group, substituted aromatic heterocyclic ring or unsubstituted or substituted or unsubstituted aliphatic heterocyclic ring; X is CH2, CHF, CF2, CHOH, S or O; n is 1, 2 or 3; and p1 is a protecting group of an amino group.

3. A pharmaceutical product, characterized in that it contains as an active ingredient, the bicycloester derivative according to claim 1, or a pharmaceutically acceptable salt thereof.

4. A DPP-IV inhibitor, characterized in that it contains as an active ingredient, the bicycloester derivative according to claim 1, or a pharmaceutically acceptable salt thereof.

5. A therapeutic agent for a disease, characterized in that it involves DPP-IV which contains as an active ingredient the bicycloester derivative according to claim 1 or a pharmaceutically acceptable salt thereof.

6. The therapeutic agent according to claim 5, characterized in that the disease involving DPP-IV is diabetes or an associated complication.