JP2005533771A - Fluorinated cyclic amides as dipeptidyl peptidase IV inhibitors - Google Patents

Abstract

The present invention relates to novel therapeutically active selective inhibitors of the enzyme dipeptidyl peptidase IV, pharmaceutical compositions comprising said compounds and type 2 diabetes mellitus, hyperglycemia, impaired glucose tolerance, metabolic syndrome (syndrome X) Or insulin resistance syndrome), grape diabetes, metabolic acidosis, cataract, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, type 1 diabetes, obesity, obesity-induced condition, hypertension , Hyperlipidemia, atherosclerosis, osteoporosis, osteopenia, fragility, bone loss, fracture, acute coronary syndrome, infertility due to polycystic ovary syndrome, short bowel syndrome, anxiety, depression, insomnia, chronic Diseases with proteins treated by DPP-IV such as fatigue, epilepsy, eating disorders, chronic pain, drinking alcohol, bowel movement related diseases, ulcers, irritable bowel syndrome, inflammatory bowel syndrome To treat, and to the use of such compounds in order to prevent disease progression in type 2 diabetes. The invention further relates to a method of identifying an insulin secretagogue for diabetes.

Description

  The present invention relates to a novel selective inhibitor having therapeutic activity for the enzyme dipeptidyl peptidase IV (hereinafter referred to as “DPP-IV”), a pharmaceutical composition containing the compound, and type 2 diabetes, metabolic syndrome (X Syndrome or insulin resistance syndrome), hyperglycemia, impaired glucose tolerance, grape diabetes, metabolic acidosis, cataract, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, type 1 diabetes, obesity , Hypertension, hyperlipidemia, atherosclerosis, osteoporosis, osteopenia, frailty, bone loss, fracture, acute coronary syndrome, infertility due to polycystic ovary syndrome, short bowel syndrome, anxiety, depression Treated with DPP-IV such as illness, insomnia, chronic fatigue, epilepsy, eating disorders, chronic pain, drinking habits, bowel movement related diseases, ulcers, irritable bowel syndrome, inflammatory bowel syndrome For treating a disease associated with that protein, and the use of such compounds for preventing disease progression in type 2 diabetes. The invention further relates to a method for identifying insulin secretagogues for diabetes.

  Dipeptidyl peptidase IV (EC 3.4.14.5) is a serine protease that preferentially hydrolyzes the N-terminal dipeptide from a protein having a proline or alanine at the 2-position. Although the physiological role of DPP-IV has not been fully elucidated, it is involved in diabetes, glucose tolerance, obesity, appetite regulation, lipemia, osteoporosis, neuropeptide metabolism and T cell activity. Conceivable.

  Since the substrate contains incretin peptide glucagon-like peptide 1 (GLP-1) and gastric acid secretion inhibitory polypeptide (GIP), DPP-IV is involved in the regulation of glucose homeostasis. When the N-terminal amino acid is cleaved from these peptides, they become functionally inactive. GLP-1 has been shown to be effective in anti-diabetic therapy for patients with type 2 diabetes and to reduce diet-related insulin requirements in patients with type 1 diabetes. GLP-1 and / or GIP are thought to regulate satiety, lipemia and bone formation. Outpatient GLP-1 has been proposed as a therapeutic for patients suffering from acute coronary syndrome, tonsillitis and ischemic heart disease.

  Administration of DPP-IV inhibitors in vivo prevents the N-terminal degradation of GLP-1 and GIP, resulting in higher circulating concentrations of these peptides, increased insulin secretion, and improved glucose tolerance. Based on these observations, DPP-IV inhibitors are considered as drugs for treating type 2 diabetes and diseases involving glucose tolerance. In addition, by treating with DPP-IV inhibitors, neuropeptide Y (NPY), peptides associated with various central nervous system diseases and gastrointestinal conditions such as ulcers, irritable bowel syndrome and inflammatory bowel syndrome Degradation of peptide YY related to is prevented.

  Insulin was discovered early and subsequently widely used in the treatment of diabetes, and sulfonylureas (eg chlorpropamide (Pfizer), tolbutamide (Upjohn), acetohexamide (EI Lilly)), biguanides (Phenformin (Ciba Geigy), metformin (GD. Seale)) and thiazolidinedione (rosiglitazone) (GlaxoSmithKline, Bristol-MyersSquibb), pioglitazone (pioglitazone (pi.). Despite being later discovered and used as an oral hypoglycemic agent, the treatment of diabetes still remains inadequate.

  The use of insulin required in type 1 diabetics and about 10% of type 2 diabetics for whom currently available oral hypoglycemic drugs are ineffective, usually requires multiple daily doses by self-injection. To do. Determining an appropriate dose of insulin requires frequent estimation of glucose concentrations in urine or blood. Administering excessive doses of insulin induces hypoglycemia, with the consequences of mild blood glucose abnormalities, coma or even death.

  Treatment of type 2 diabetes usually involves a combination of therapeutic diet, exercise, oral medication, and, in more severe cases, insulin. However, clinically available hypoglycemic drugs may exhibit side effects that limit their use. It is clear that there is a continuing need for hypoglycemic drugs that have fewer side effects or can be successful if other drugs fail.

  Inadequately controlled hyperglycemia is a direct cause of numerous complications that characterize advanced diabetes mellitus (cataracts, neurosis, nephropathy, retinopathy, cardiomyopathy). In addition, diabetes mellitus is a comorbid disease often confused with hyperlipidemia, atherosclerosis and hypertension, which significantly increases the overall morbidity and overall mortality from these diseases.

  Epidemiological evidence confirms hyperlipidemia as a major risk factor for cardiovascular disease (“CVD”) due to atherosclerosis. Atherosclerosis is recognized as a leading cause of death in the United States and Western Europe. At least in part, CVD is particularly widespread in diabetic patients because there are multiple independent risk factors such as glucose intolerance, left ventricular hypertrophy and hypertension in this population. Therefore, successful treatment of hyperlipidemia in the normal population, especially in diabetic patients, is of great medical importance.

  Hypertension (or high blood pressure) is a condition that can occur in many patients whose causative factors or diseases are not known. Such “essential” hypertension is often associated with diseases such as obesity, diabetes and hypertriglyceridemia, and hypertension is known to be positively associated with heart failure, renal failure and stroke. Yes. Hypertension can also contribute to the development of atherosclerosis and coronary artery disease. Hypertension includes a group of symptoms that characterize the metabolic syndrome, also known as insulin resistance syndrome ("IRS") and syndrome X, along with insulin resistance and hyperlipidemia.

  Obesity is a well-known common risk factor for the development of atherosclerosis, hypertension and diabetes. The incidence of obesity and hence the incidence of these diseases is increasing worldwide. Currently, few drugs that are effective and tolerably reduce obesity are available.

  Osteoporosis is a progressive systemic disease characterized by low bone density and microstructural alteration of bone tissue, resulting in increased bone fragility and ease of fracture. The consequences of osteoporosis and impaired bone strength are obvious causes of vulnerability and high morbidity and mortality.

  Heart disease is a major health problem worldwide. Myocardial infarction is a significant cause of death for individuals with heart disease. Acute coronary syndrome refers to patients who have or have a high risk of developing acute myocardial infarction (MI).

  Although there are therapies that can be used to treat diabetes, hyperglycemia, hyperlipidemia, hypertension, obesity and osteoporosis, the need for alternative improved therapies continues to exist.

  The present invention relates to type 2 diabetes, metabolic syndrome (syndrome X or insulin resistance syndrome), hyperglycemia, impaired glucose tolerance, grape diabetes, metabolic acidosis, cataract, diabetic neuropathy, diabetic nephropathy, diabetic Due to retinopathy, diabetic cardiomyopathy, type 1 diabetes, obesity, hypertension, hyperlipidemia, atherosclerosis, osteoporosis, osteopenia, fragility, bone loss, fracture, acute coronary syndrome, polycystic ovary syndrome Treats infertility, short bowel syndrome, anxiety, depression, insomnia, chronic fatigue, epilepsy, eating disorders, chronic pain, drinking habits, bowel movement related diseases, ulcers, irritable bowel syndrome, inflammatory bowel syndrome In order to do and to prevent disease progression in type 2 diabetes, compounds of formula I are directed.

  In particular, the present invention is directed to a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug,

[In the above formula,
R ¹ is 3-fluoroazetidin-1-yl, 3,3-difluoroazetidin-1-yl, 3,4-difluoropyrrolidin-1-yl, 3,3,4-trifluoropyrrolidin-1-yl 3,3,4,4-tetrafluoropyrrolidin-1-yl, 3-fluoropiperidin-1-yl, 4-fluoropiperidin-1-yl, 3,4-difluoropiperidin-1-yl, 3,5- Difluoropiperidin-1-yl, 3,3-difluoropiperidin-1-yl, 4,4-difluoropiperidin-1-yl, 3,4,5-trifluoropiperidin-1-yl, 3,3,4-tri Fluoropiperidin-1-yl, 3,3,5-trifluoropiperidin-1-yl, 3,4,4-trifluoropiperidin-1-yl, 3,3,4,5-tetrafluoropipe Lysine-1-yl, 3,4,4,5-tetrafluoropiperidin-1-yl, 3,3,4,4-tetrafluoropiperidin-1-yl, 3,3,5,5-tetrafluoropiperidine- 1-yl, 3,3,4,5,5-pentafluoropiperidin-1-yl, 3,3,4,4,5-pentafluoropiperidin-1-yl or 3,3,4,4,5 5-hexafluoropiperidin-1-yl, and R ² is (C ₁ -C ₈ ) alkyl or (C ₃ -C ₈ ) cycloalkyl].

In a preferred embodiment, the compound of formula I is a compound wherein R ¹ is 3-fluoroazetidin-1-yl or 3,3-difluoroazetidin-1-yl.

In other preferred embodiments, the compound of Formula I is such that R ¹ is 3,4-difluoropyrrolidin-1-yl, 3,3,4-trifluoropyrrolidin-1-yl, 3,3,4,4- It is a compound that is tetrafluoropyrrolidin-1-yl.

In other preferred embodiments, the compound of Formula I is such that R ¹ is 3-fluoropiperidin-1-yl, 4-fluoropiperidin-1-yl, 3,4-difluoropiperidin-1-yl, 3,5- A compound that is difluoropiperidin-1-yl, 3,3-difluoropiperidin-1-yl or 4,4-difluoropiperidin-1-yl.

In other preferred embodiments, the compound of Formula I is such that R ¹ is 3,4,5-trifluoropiperidin-1-yl, 3,3,4-trifluoropiperidin-1-yl, 3,3,5 -Trifluoropiperidin-1-yl, 3,4,4-trifluoropiperidin-1-yl, 3,3,4,5-tetrafluoropiperidin-1-yl, 3,4,4,5-tetrafluoropiperidine -1-yl, 3,3,4,4-tetrafluoropiperidin-1-yl, 3,3,5,5-tetrafluoropiperidin-1-yl, 3,3,4,5,5-pentafluoropiperidine It is a compound that is -1-yl, 3,3,4,4,5-pentafluoropiperidin-1-yl or 3,3,4,4,5,5-hexafluoropiperidin-1-yl.

In another preferred embodiment, the compound of formula I is
Hydrochloric acid (2S, 3S) -2-amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -pentan-1-one,
Hydrochloric acid (2S, 3S) -2-amino-1- (3-fluoro-azetidin-1-yl) -3-methyl-pentan-1-one,
Hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -ethanone,
Hydrochloric acid (2S, 3R) -2-amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -pentan-1-one,
Hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (3-fluoro-azetidin-1-yl) -ethanone,
Hydrochloric acid (S) -2-amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -butan-1-one;
Hydrochloric acid (S) -2-amino-4-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -pentan-1-one;
Hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (3,3-difluoro-azetidin-1-yl) -ethanone,
Hydrochloric acid (2S, 3S) -2-amino-1- (3,3-difluoro-azetidin-1-yl) -3-methyl-pentan-1-one,
Hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (4,4-difluoro-piperidin-1-yl) -ethanone,
Hydrochloric acid (2S, 3S) -2-amino-1- (4,4-difluoro-piperidin-1-yl) -3-methyl-pentan-1-one,
Hydrochloric acid (S) -2-amino-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -propan-1-one;
Hydrochloric acid (S) -2-amino-1- (3,3-difluoro-azetidin-1-yl) -3-methyl-butan-1-one;
Hydrochloric acid (S) -2-amino-1- (3-fluoro-azetidin-1-yl) -3-methyl-butan-1-one;
Hydrochloric acid (2S, 3R) -2-amino-1- (3-fluoro-azetidin-1-yl) -3-methyl-pentan-1-one,
Hydrochloric acid (2S, 3R) -2-amino-1- (3,3-difluoro-azetidin-1-yl) -3-methyl-pentan-1-one,
Hydrochloric acid (S) -2-amino-2-cyclopentyl-1- (3,3-difluoro-azetidin-1-yl) -ethanone,
Hydrochloric acid (S) -2-amino-2-cyclopentyl-1- (3-fluoro-azetidin-1-yl) -ethanone or hydrochloric acid (S) -2-amino-2-cyclopentyl-1- (3,3,4 , 4-tetrafluoro-pyrrolidin-1-yl) -ethanone.

  In another preferred embodiment, the compound of formula I is

  S enantiomer shown as

  In another aspect, the invention provides a therapeutically effective amount of a compound of formula I as described above, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug and a pharmaceutically acceptable diluent or A pharmaceutical composition comprising a carrier is intended.

In yet another aspect, the invention provides a therapeutically effective amount of a first compound of formula I as defined above, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug and insulin or an insulin analog. Insulinotropin; biguanide; α ₂ -antagonist or imidazoline; glitazone; aldose reductase inhibitor; glycogen phosphorylase inhibitor; sorbitol dehydrogenase inhibitor; fatty acid oxidation inhibitor; α-glucosidase inhibitor; -Agonists; phosphodiesterase inhibitors; lipid-lowering agents; anti-obesity agents; vanadate, vanadium complexes or peroxovanadium complexes; amylin antagonists; glucagon antagonists; Gluconeogenesis inhibitor; somatostatin analog; renal glucose inhibitor; or second compound that is an antilipolytic agent; prodrug of said second compound or said second drug It is intended for a pharmaceutical composition containing a compound or a pharmaceutically acceptable salt of a prodrug of said second compound. In other embodiments of this aspect of the invention, the composition further comprises a pharmaceutically acceptable carrier or diluent.

In another aspect, the present invention provides:
a) a first dosage form comprising a compound of formula I as defined above;
b) Insulin and insulin analogues; insulinotropin; biguanides; alpha ₂ - antagonists and imidazolines; glitazones; aldose reductase inhibitors; glycogen phosphorylase inhibitor; sorbitol dehydrogenase inhibitors; fatty acid oxidation inhibitors; alpha-glucosidase inhibitor; beta -Agonists; phosphodiesterase inhibitors; lipid-lowering agents; anti-obesity agents; vanadate and vanadium and peroxovanadium complexes; amylin antagonists; glucagon antagonists; growth hormone secretagogues; gluconeogenesis inhibitors; A second dosage form comprising an anti-diabetic agent selected from an inhibitor; or an anti-lipolytic agent; a prodrug of this second dosage form and a pharmaceutically acceptable of the second dosage form and the prodrug To a kit comprising a container means for including a and c) said first dosage form (a) and said second dosage form (b) the target; kill salts.

  In one embodiment, the kit further comprises a pharmaceutically acceptable carrier or diluent and container means for these carriers or diluents.

  In another aspect, the present invention is directed to a therapeutic method for inhibiting dipeptidyl peptidase IV in a mammal, which method comprises a therapeutically effective amount of a compound of formula I, a prodrug thereof or the compound or Administering a pharmaceutically acceptable salt of a prodrug of the above or a pharmaceutical composition as described above to said mammal in need of inhibiting dipeptidyl peptidase IV.

  In another aspect, the present invention is directed to a method of treating a condition mediated by dipeptidyl peptidase-IX inhibition in a mammal, said method comprising a therapeutically effective amount of a compound of formula I, Administering a drug or a pharmaceutically acceptable salt of said compound or said prodrug or said pharmaceutical composition to said mammal suffering from a condition alleviated by dipeptidyl peptidase-IX inhibition. Including.

  In one embodiment of this method, the condition being treated is type 2 diabetes, metabolic syndrome, hyperglycemia, impaired glucose tolerance, grape diabetes, metabolic acidosis, cataracts, diabetic neuropathy, diabetic nephropathy, diabetes Retinopathy, diabetic cardiomyopathy, type 1 diabetes, obesity, condition worsened by obesity, hypertension, hyperlipidemia, atherosclerosis, osteoporosis, osteopenia, fragility, bone loss, fracture, acute coronary Syndrome, infertility due to polycystic ovary syndrome, disease progression in type 2 diabetes, chronic fatigue, epilepsy, diseases related to bowel movement, ulcer, irritable bowel syndrome, inflammatory bowel syndrome, anxiety, depression, insomnia, Chronic fatigue, epilepsy, eating disorders, chronic pain, or drinking habits.

  In a preferred embodiment, the condition being treated is type 2 diabetes.

In another aspect, the invention is directed to a method of identifying whether an agent is a DPP-IV inhibitor, the method comprising:
a) administering the drug to a fasted diabetic KK / H1J mouse;
b) loading KK / H1J mice with oral glucose;
c) assessing the response of the mouse to a subsequent oral glucose load, wherein said agent is treated with type 2 diabetes, metabolic syndrome, hyperglycemia, impaired glucose tolerance, grape diabetes, metabolic acidosis, Cataract, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, type 1 diabetes, obesity, condition exacerbated by obesity, hypertension, hyperlipidemia, atherosclerosis, osteoporosis, Osteopenia, fragility, bone loss, fracture, acute coronary syndrome, infertility due to polycystic ovary syndrome, prevention of disease progression in type 2 diabetes, anxiety, depression, insomnia, chronic fatigue, epilepsy, eating disorders, It can be identified as a treatment for chronic pain, alcohol drinking, intestinal motility related diseases, ulcers, irritable bowel syndrome, inflammatory bowel syndrome or short bowel syndrome.

  As used herein, the term “alkyl” refers to a monovalent saturated straight or branched chain aliphatic hydrocarbon group having from 1 to 8 carbon atoms.

As used herein with respect to the compounds of formula I of the present invention, the expression “pharmaceutically acceptable salt” includes pharmaceutically acceptable anionic salts. The term “pharmaceutically acceptable anion” refers to an anion that is chemically and / or toxically applicable to other components of the pharmaceutical composition and / or the animal treated therewith. Suitable anions include, but are not limited to, halides (eg, chloride, iodo and bromide), (C ₁ -C ₁₂ ) alkyl sulfonates (eg, mesylate, ethyl sulfonate, etc.), aryl sulfonates (eg, phenyl sulfonate) , Tosylate, etc.), (C ₁ -C ₁₂ ) alkyl phosphonate, di (C ₁ -C ₁₂ ) alkyl phosphate (eg dimethyl phosphate, diethyl phosphate, α-diglycerol phosphate, etc.), aryl phosphonate, aryl phosphate, alkyl aryl phosphonates, alkyl aryl _phosphates, (C 1 _{-C 12)} alkyl carboxylate (e.g., acetate, propionate, glutamate, etc. glycerate), aryl carboxylate Etc. are included.

  The compounds of the present invention can be isolated or used as such or in the form of pharmaceutically acceptable salts, solvates and / or hydrates. The term “salt” means inorganic and organic salts of the compounds of the invention. These salts are prepared by reacting the compound or prodrug with an appropriate organic or inorganic acid in situ, or separately during final isolation and purification of the compound, and isolating the resulting salt. be able to. Typical salts include hydrobromide, hydrochloride, hydrogen iodide, sulfate, bisulfate, nitrate, acetate, trifluoroacetate, oxalate, besylate, palmitate, pamo Acid salt, malonate, stearate, laurate, malate, borate, benzoate, lactate, phosphate, hexafluorophosphate, benzenesulfonate, tosylate, formic acid Citric acid, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptate, lactobionate and lauryl sulfonate. For example, Berge et al. Pharm. Sci. 66, 1-19 (1977).

  The term “prodrug” means a compound that is transformed in vivo to yield a compound of formula (I) or a pharmaceutically acceptable salt, hydrate or solvate of the compound. Such compounds include, but are not limited to, N-acyl and N-carboalkoxy derivatives of compounds of formula I, as well as imine derivatives. Transformation can occur through various mechanisms, such as through hydrolysis in blood. For a discussion on the use of prodrugs, see T.W. Higuchi and W.H. Stella's “Pro-drugs as Novel Delivery Systems”, Vol. 14 of the A.E. C. S. Symposium Series and Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.

Some of the compounds described herein contain at least one asymmetric center and, therefore, those skilled in the art will recognize all stereoisomers of the compounds described and discussed herein ( For example, it will be appreciated that enantiomers and diastereoisomers and racemic mixtures thereof are within the scope of the invention. In particular, the carbon substituted with R ⁷ in the compound of formula I and its intermediates is asymmetric and these compounds are claimed in claim 1 as racemic mixtures and in claim 6 as S enantiomers. Shown and claimed. One skilled in the art will recognize that any of the pyrrolidine and piperidine groups of the compound of Formula I may contain at least one asymmetric center (eg, Example 7). All stereoisomers (eg, enantiomers, diastereomers, and racemic mixtures thereof) of these compounds that are claimed, depicted and discussed herein are within the scope of the invention.

  In addition, those skilled in the art will understand that compounds of formula I are in crystalline form as hydrates in which water molecules are contained within the crystal structure and as solvates in which solvent molecules are contained. I will admit that it can exist. All such hydrates and solvates are considered part of this invention.

Furthermore, the present invention is the same as the compounds represented by formula I, but in fact one or more atoms have an atomic mass or mass number different from the atomic mass or mass number normally present in nature. Also included are isotopically-labelled compounds that have been replaced by Examples of isotopes that can be introduced into the compounds of the invention include hydrogen, carbon, nitrogen, oxygen and fluorine, such as ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O and ¹⁸ F, respectively. Isotopes. Compounds of the present invention, prodrugs thereof, pharmaceutically acceptable salts of the present compounds or prodrugs that contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of the present invention. Certain isotopically-labelled compounds of the present invention, for example those into which radioactive isotopes such as ³ H and ¹⁴ C have been introduced, can be used in drug and / or substrate tissue distribution analyses. Due to their ease of preparation and detectability, tritium labels (ie, ³ H) and carbon-14 (ie, ( ¹⁴ C) isotopes are particularly preferred, and are heavier, such as deuterium (ie, ² H). Isotope substitution can result in certain therapeutic advantages resulting from greater metabolic stability, eg, high in vivo half-life or low dose requirements, and may be advantageous in some cases. The compounds of formula I and their prodrugs are typically subjected to the procedures discussed in the schemes and / or examples below, replacing unisotopically labeled reagents with readily available isotope labeled reagents Can be prepared.

  In general, the compounds of formula I of the present invention can be prepared by methods involving processes known in the chemical arts, particularly in light of the description contained herein. One process for preparing the compounds of formula I of the present invention is detailed by the following reaction scheme. Other processes are described in the Examples section. Several of the starting compounds for the reactions described in the schemes and examples are prepared as detailed herein. All other starting compounds are available from common commercial sources such as Sigma-Aldrich Corporation (St. Louis, MO).

In Scheme 1, (Step 1) an amino acid compound of Formula II wherein R ¹⁰ is a nitrogen protecting group compatible with Chemical Scheme I above (eg, (L) -Boc-isoleucine, (L ) -Boc-cyclohexylglycine, (L) -Boc-allo-isoleucine, (L) -Boc-leucine, (L) -Boc-valine or (L) -Boc-alanine) and a compound of formula III, fluorination R ¹ , R ² , R ³ , R ⁴ and R ^{7 in the} formula by coupling with pyrrolidine (eg 3,3-difluoropyrrolidine hydrochloride or 3,3,4,4-tetrafluoropyrrolidine hydrochloride) Compounds of formula I can be prepared wherein are defined as above.

Suitable nitrogen protecting groups R ¹⁰ include, but are not limited to, for example, t-butoxycarbonyl (“Boc”), benzyloxycarbonyl, and fluorenylmethoxycarbonyl (“Fmoc”). Other examples of nitrogen protecting groups are described in “Protective Groups in Organic Synthesis”, 2nd. Ed. , P.M. G. M.M. Wuts and T.W. W. Green, page 315. Coupling with a compound of formula II yields a compound of formula IV. In step 2, the compound of formula IV is dissolved in an inert solvent (eg, ethyl acetate) and a method suitable for the nature of the R ¹⁰ group (eg, if R ¹⁰ is Boc, as described in the above references). Deprotection with a gaseous acid) gives compounds of formula I.

  The coupling reaction can be easily performed by dissolving the compound of formula II and the compound of formula III in a reaction inert solvent (eg, dichloromethane) in the presence of a base (eg, triethylamine or pyridine). To the resulting solution is added a coupling agent or combination of coupling agents (eg, hydroxybenzotriazole / hydrochloric acid 1-(-3-dimethylaminopropyl) -3-ethylcarbodiimide). 1-hydroxy-7-azabenzotriazole / hydrochloric acid 1-(-3-dimethylaminopropyl) -3-ethylcarbodiimide, hydroxybenzotriazole / dicyclohexylcarbodiimide, 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline, Other coupling agents such as carbonyldiimidazole or diethyl phosphoryl cyanide can also be used. Coupling is performed in an inert solvent, preferably an aprotic solvent. Suitable solvents include, for example, acetonitrile, dichloromethane, dimethylformamide, chloroform. To examine other conditions that can be used to couple carboxylic acids, see Houben-Weyl, Vol XV, part. 11, E.M. Wunsch, Ed. G. Theime Verlag, 1974, Stuttgart and M.C. Bodansky, Principles of Peptide Synthesis (SpringerVerlag Berlin 1984) and The peptides. See Analysis, Synthesis and Biology (ed. E. Gross and J. Meienhofer), vols 1-5 (Academic Press NY 1979-1983). The texts of the above references are incorporated by reference.

  The reaction is typically carried out at ambient pressure and temperature until the starting material is no longer present as detected by thin layer chromatography or other analytical techniques well known to those skilled in the art. The coupling product of formula IV can be isolated by methods well known to those skilled in the art.

  Decomposition of the compound of formula IV by dissolving the compound of formula IV in an inert solvent (eg, ethyl acetate), cooling to about 0 ° C., followed by treatment with a gaseous acid (eg, hydrochloric acid) for about 1 minute. Protection can be easily performed. The reaction mixture is stirred for about 15 minutes, then allowed to reach room temperature, followed by stirring for an additional about 30 minutes.

  One skilled in the art can replace the protected (L) amino acid compound of formula II shown in Scheme III and illustrated in Examples 2-8 with a racemic mixture of compounds of formula II. Would admit. Accordingly, compounds of formula I may exist as racemic mixtures of enantiomers and these mixtures are within the scope of the invention.

  As used herein, the term “inert solvent” is a solvent whose structure does not contain a functional group that would interfere with the reaction. Examples for activating and coupling the hydroxyl group are dichloromethane, 1,2-dichloroethane, tetrahydrofuran (THF), dimethylformamide (DMF).

  As used herein, the term “activating reagent” in this example is to change a hydroxyl group to a leaving group such as bromide, iodo, alkyl sulfonate or aryl sulfonate.

  Optically active amino acids can be obtained prior to coupling in Step 1 of Scheme III by degradation, by asymmetric synthesis, or by other methods well known to those skilled in the art. Alternatively, if desired, degradation can be performed at a later point in the synthesis of the compound of formula I by techniques known to those skilled in the art.

  Compound III of Scheme I can be prepared as known to one of ordinary skill in the art. See, for example, Giardina, G et al., Svnlet. It is also possible to prepare 3,3-difluoropyrrolidine hydrochloride as described in 1995, page 55. Chaudry et al. Chem. Soc. 1964, p. 874, 3,3,4,4-tetrafluoropyrrolidine can also be prepared. Alternatively, 3,3,4,4-tetrafluoropyrrolidine can be prepared as shown in Scheme II below.

In Scheme II, in Step 1, the hydroxyl group of 2,2,3,3-tetrafluorobutanediol (X) is activated to leave the leaving group R ⁸ (where R ⁸ is Br, I or SO). ₂ R ¹¹ , where R ¹¹ is (1) a C ₁ -C ₈ linear or branched alkyl optionally substituted with one or more fluorines or (2) halogen. May be an aryl group optionally substituted by or a C ₁ -C ₈ linear or branched alkyl optionally substituted by 1 or 4 fluorines). Preferably, however, R ⁸ is a trifluoromethylsulfonyloxy group (F ₃ CSO ₃ ). Activating reagents necessary to convert alcohol functionality to, for example, fluorine or iodine are well known to those skilled in the art (eg, March, Advanced Organic Chemistry, 3rd ed., Pages 382-384, incorporated by reference). and Larock, in Comprehensive Organic Transformations, pp. 353-360), but are not limited to, the following activating reagents: hydrogen bromide ( "HBr"), phosphorus tribromide ( "PBr _3"), pentabromide Phosphorus (“PBr ₅ ”), thionyl bromide (“SOBr ₂ ”) and hydrogen iodide (“HI”) are included. Preferred activating reagent combinations are triphenylphosphine / carbon tetrabromide (“Ph ₃ P / CBr ₄ ”), Ph ₃ P / N-bromosuccinimide, potassium iodide / phosphoric acid (“KI / H ₃ PO _4). )), Ph ₃ P / I ₂ and Me ₃ SiCl / Nal. Activating an alcohol function to an alkyl or aryl sulfonate involves reacting the corresponding sulfonyl chloride or sulfonic anhydride with an inert solvent in the presence of a base such as pyridine or triethylamine and cooling it (0 C.) and stirring for about 1 hour. The reaction mixture is then stirred at room temperature for about an additional hour.

In Step 2, a compound XI and a primary amine R ⁹ NH ₂ where R ⁹ is an alkyl or aryl “protecting group” (ie, the bond between R ⁹ and the nitrogen atom is known to those skilled in the art in Step 3. Reaction can be easily separated by standard chemical treatment) by heating the solution to reflux overnight. Examples of N-protecting groups are described in “Protective Groups in Organic Synthesis”, 2nd. Ed. , P.M. G. M.M. Wuts and T.W. W. Greene, page 362, and includes, for example, benzyl, t-butyl, allyl and benzhydryl. Preferably R ⁹ is benzyl, in which case the deprotection of step 3 is carried out by hydrogenolysis in the presence of palladium.

Step 3 can be performed with a cationic salt of intermediate compound XII, such as hydrochloride, hydrobromide, acetate, trifluoroacetate, etc., in a suitable solvent (eg, water, methanol or ethanol). Removal of the R ⁹ protecting group from compound XII can be performed under conditions appropriate for the particular R ⁹ protecting group used. Such conditions include, for example, (a) hydrogenolysis when R ⁹ is benzyl or benzhydryl, (b) treatment with a strong acid such as trifluoroacetic acid or hydrochloric acid when R ⁹ is t-butyl. Or (c) treatment with tributyltin hydride and acetic acid in the presence of a catalytic amount of bis (triphenylphosphine) palladium (II) when R ⁹ is allyl.

When R ⁹ is benzyl, deprotection is carried out by hydrogenolysis for about 3 hours at about 45 psi in the presence of 10% palladium in ethanol. Filtration of the catalyst through diatomaceous earth, removal of the solvent and trituration with a non-hydroxyl solvent such as diethyl ether, diisopropyl ether, ethyl acetate, 1,4-dioxane or tetrahydrofuran, then yields the final compound III as the corresponding cationic salt. Isolate. As described in Scheme I, Compound III is utilized to prepare DPP-IV inhibitor compounds of Formula I.

Alternatively, compounds of formula I where a and b are each 0 can be prepared according to the steps of Scheme III. In Step 1 of Scheme III, an (L) amino acid compound of Formula II (eg, (L) -Boc-isoleucine) in which R ⁷ and R ¹⁰ are defined as above is used in Step 1 of Scheme I. As above, coupling with hydroxyazetidine (compound of formula V) yields the compound of formula VI. Treatment of the resulting compound of formula VI in step 2 with diethylaminosulfur trifluoride or similar fluorinating agent provides the compound of formula VII. In Step 3, the compound of Formula VII is treated with an acid as described above in Step 2 of Scheme I to give a compound of Formula I where a and b are each 0.

  A solution of diethylaminosulfur trifluoride in a reaction inert solvent (eg dichloromethane) is cooled (eg −78 ° C.) and to this is added dropwise a solution of the compound of formula VI. The reaction can be performed easily. The reaction mixture is warmed to ambient temperature until no more starting material is present or the reaction is complete, as determined by thin layer chromatography or other analytical techniques well known to those skilled in the art. The compound of formula VII can be isolated by methods well known to those skilled in the art and further treated with a gaseous acid in step 3 as described above in step 2 of Scheme I.

  Similarly, monofluoropyrrolidine and piperidine derivatives can be prepared in a similar order, starting from pyrrolidin-3-ol or hydroxypiperidine, respectively.

Alternatively, compounds of formula I can be prepared according to Scheme IV. In step 1, as described above in step 1 of scheme 1, the (L) amino acid compound of formula II wherein R ⁷ and R ¹⁰ are as defined above (eg (L) -Boc-isoleucine , Boc-cyclohexylglycine, etc.), a compound of formula VIII, wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are as defined above, a fluorinated piperidine (eg, hydrochloric acid 4 , 4-difluoro-piperidine) to give compounds of formula IX. As described above in Step 2 of Scheme 1, deprotection of the compound of Formula IX in Step 2 (eg, a gaseous acid) provides the compound of Formula I.

  One skilled in the art can also apply the synthetic procedures used in Schemes I and 4 to the synthesis of similar compounds starting from the requisite azetidine starting material and containing an azetidine ring instead of a pyrrolidine or piperidine group. Would admit.

  One of ordinary skill in the art will know the protected (L) amino acid compound of formula II shown in Schemes 1, 3 and 4 and illustrated in Examples 1-16, of formula II It will be appreciated that a racemic mixture of the following compounds can be substituted. Accordingly, compounds of formula I may exist as racemic mixtures of enantiomers and these mixtures are within the scope of the invention.

  Obtaining an optically active amino acid prior to coupling in Step 1 of Schemes I, III and IV by optical resolution, by asymmetric synthesis, or by other methods well known to those skilled in the art. it can. Alternatively, if desired, optical resolution can be performed at a later point in the synthesis of a compound of formula I by techniques known to those skilled in the art.

  The compounds of formula I of the present invention are used to treat dipeptidyl peptidase IV related conditions; treat type 2 diabetes; prevent disease progression in type II diabetes; type I diabetes, impaired glucose tolerance, hyperglycemia, impaired glucose tolerance, Metabolic syndrome (syndrome X or insulin resistance syndrome), grape diabetes, metabolic acidosis, cataract, diabetic neuropathy and nephropathy, metabolic syndrome, treatment of obesity, hypertension, hyperlipidemia, metabolic acidosis, arthritis, Osteoporosis, osteopenia, fragility, bone loss, fracture, short stature due to lack of growth hormone, infertility due to multiple cyst-ovary syndrome, anxiety, depression, insomnia, chronic fatigue, epilepsy, eating disorders, chronic pain Useful for the treatment of drunk, intestinal motility related diseases, ulcers, irritable bowel syndrome, inflammatory bowel syndrome and short bowel syndrome.

  The present invention further relates to a therapeutic method for treating or preventing said condition in mammals, including humans, wherein a compound of formula I of the present invention is administered in an appropriate dosage regimen designed to produce a therapeutic effect. Administer as part. The appropriate dosage regimen, the amount of each dose administered and the interval between doses of the compound will depend on the compound of formula I of the invention used, the type of pharmaceutical composition used, the characteristics and condition of the patient being treated. Depends on severity.

  In general, an effective dose with a compound of formula I is 0.01 mg / kg / day to 30 mg / kg / day, preferably 0.01 mg / kg / day to 1 mg / day, in one or more administrations. kg / day. However, depending on the condition of the patient being treated, some variation of this dose will inevitably occur. In either case, the individual responsible for determining the dose will determine the appropriate dose for the individual patient.

  The compounds of formula I of the present invention can be administered to a patient in need of treatment by a variety of conventional routes of administration, including oral and parenteral (eg, intravenous, subcutaneous or intramedullary). Furthermore, the pharmaceutical composition of the present invention is administered intranasally, as a suppository, or using a “flash” formulation, ie, dissolving the pharmaceutical in the mouth without the need for water. be able to.

  The compounds of the invention can be administered in a single dose (eg, once a day) or in multiple doses, or by constant infusion. The compounds of the present invention can also be administered alone or in combination with a pharmaceutically acceptable carrier, vehicle or diluent in one or more administrations. Suitable pharmaceutical carriers, vehicles and diluents include inert solid diluents or fillers, sterile aqueous solution and various organic solvents. The pharmaceutical composition resulting from the combination of a compound of the present invention and a pharmaceutically acceptable carrier, vehicle or diluent is then readily available in various dosage forms such as tablets, powders, troches, syrups, injectable solutions and the like. Can be administered. If desired, these pharmaceutical compositions may contain other ingredients such as perfumes, binders, excipients and the like.

  Thus, for oral administration purposes, tablets containing various excipients such as sodium citrate, calcium carbonate and / or calcium phosphate are combined with various disintegrants such as starch, alginic acid and / or certain complex silicates. , Polyvinylpyrrolidone, sucrose, gelatin and / or a binder such as gum arabic. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate and talc often serve tableting purposes. Similar types of solid compositions can also be used as fillers in soft and hard filled gelatin capsules. Preferred materials for this include lactose or milk sugar and high molecular weight polyethylene glycols. If an aqueous suspension or elixir is desired for oral administration, the active agent in it may be mixed with various sweetening or flavoring agents, coloring agents or dyes, if desired emulsifying agents or suspending agents, and water. Can be combined with diluents such as ethanol, propylene glycol, glycerin and / or combinations thereof.

  For parenteral administration, solutions of the compounds of the invention in sesame oil, peanut oil, aqueous propylene glycol, or in sterile aqueous solution can be used. Such aqueous solutions should be appropriately buffered, if necessary, and the liquid diluent first made isotonic with sufficient saline or glucose. These individual aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, the sterile aqueous medium used is readily available by standard techniques known to those skilled in the art.

  For intranasal administration or administration by inhalation, in the form of a solution or suspension from a pump spray container that is pushed or pumped by the patient, or a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane The compounds of the invention are easily delivered as an aerosol spray from a pressurized container or nebulizer using dichlorotetrafluoroethane, carbon dioxide or other suitable gas. For pressurized aerosols, the dosage unit can be determined by providing a valve to deliver a metered amount. The pressurized container or nebulizer may contain a solution or suspension of the compound of the invention. Capsules and cartridges (eg, made of gelatin) for use in an inhaler or insufflator will contain a powder mix containing one or more compounds of the invention and a suitable powder base such as lactose or starch. Can be prescribed.

Since the present invention has aspects relating to the treatment of the aforementioned indications by treating with a combination of compounds that can be co-administered separately, the present invention relates to combining separate pharmaceutical compositions into kit form. The kit comprises two separate compositions: (1) a first agent comprising a compound of formula I, a prodrug or pharmaceutically acceptable salt and prodrug thereof, and a pharmaceutically acceptable diluent or carrier. Forms; and (2) insulin and insulin analogs; insulinotropins; biguanides; α ₂ -antagonists and imidazolines; glitazones; aldose reductase inhibitors; glycogen phosphorylase inhibitors; sorbitol dehydrogenase inhibitors; Β-agonist; phosphodiesterase inhibitor; lipid-lowering agent; anti-obesity agent; vanadate, vanadium complex and peroxovanadium complex; amylin antagonist; glucagon antagonist; growth hormone secretagogue; gluconeogenesis inhibitor; A statin analog; an inhibitor of renal glucose; a second dosage form comprising an antidiabetic drug selected from antilipolytic agents; a prodrug of this second dosage form and a pharmaceutical of the second dosage form and the prodrug And a second dosage form containing a pharmaceutically acceptable carrier and a pharmaceutically acceptable carrier or diluent.

  The amounts of (1) and (2) are such that the conditions as described above are treated or ameliorated when co-administered. The kit includes a container for providing separate dosage forms, such as a separate bottle or a separate foil wrap, wherein each compartment contains multiple dosage forms (eg, tablets) containing (1) or (2) To do. Alternatively, rather than separating active ingredient-containing dosage forms, the kit may comprise separate compartments each containing an entire dosage form including separate dosage forms.

  An example of this type of kit is a blister pack, where each individual blister contains two (or more) pharmaceutical composition dosage forms from (1) and (2) pharmaceutical composition dosage forms, respectively. The above tablets are contained. The kit typically includes instructions for administering the other components. When the separate components are administered preferably in different dosage forms (eg oral and parenteral), at different dosing intervals or when titration of the individual components of the combination is directed by the prescribing physician Moreover, this kit form is particularly advantageous.

In the case of the present invention, therefore, the kit is
d) a first dosage form comprising a compound of formula I, a prodrug or pharmaceutically acceptable salt or prodrug thereof;
e) insulin and insulin analogs; insulinotropin; biguanides; alpha ₂ - antagonists and imidazolines; glitazones; aldose reductase inhibitors; glycogen phosphorylase inhibitor; sorbitol dehydrogenase inhibitors; fatty acid oxidation inhibitors; alpha-glucosidase inhibitor; beta -Agonists; phosphodiesterase inhibitors; lipid-lowering agents; anti-obesity agents; vanadate and vanadium and peroxovanadium complexes; amylin antagonists; glucagon antagonists; growth hormone secretagogues; gluconeogenesis inhibitors; A second dosage form comprising an anti-diabetic agent selected from an inhibitor; or an anti-lipolytic agent; a prodrug of this second dosage form and a pharmaceutically acceptable of the second dosage form and the prodrug Salt kill;
f) any pharmaceutically acceptable carrier or diluent; and g) said first dosage form (a), said second dosage form (b) and said optional carrier or diluent (c). Including container means.

  Methods for preparing various pharmaceutical compositions having a certain amount of active ingredient are known or will be apparent to those of skill in the art in view of the present disclosure. For examples of methods of preparing pharmaceutical compositions, see Reminton's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. 19th Edition (1995).

In Vitro Analysis for Dipeptidyl Peptidase Inhibition Dipeptidyl peptidase inhibition is a published method for measuring DPP-IV activity (Assay of dipeptidyl peptidase IV in serum by nitro-2-8. ) Sharpe, S., DeMester, I., Vanhoof, G., Hendriks, D., Van Sande, M., Van Camp, K. and Yaron, A. Clin. Chem. 34: 2299-2301; of human lymphocytes (1988) Rodja, Z. Czechoslovak Medici ne, 11: 181-194) and can be verified in vitro by the following analysis. Per 10 mL of 50 mM Gly-Pro-4-methoxy B naphthylamide HCl (eg 50 mM Tris assay buffer (pH 7.3) containing 0.1 M sodium chloride, 0.1% (v / v) Triton) Gly-Pro-4-methoxy B naphthylamide HCl (182 μg) and enzyme diluted 1: 100 (100 μL enzyme per 10 mL substrate solution) (Enzyme Systems Products Cat # SPE-01, DPP-IV 5 mU / stock) mL) to form an enzyme substrate solution, which is kept at 4 ° C. Pipet 150 μL of enzyme substrate solution into microtiter wells of polystyrene 96 well plate and maintain at 4 ° C. Add 5 μL / well of the compound of formula I to a final concentration of the compound of formula I of 3 μM to 10 nM per well.

As a control reagent blank, remove enzyme from 4 wells. 5 μL of 3 mM Diprotin A is added to 4 wells as a positive quality control to give a final diprotin A concentration of 100 μM. To measure total enzyme activity without the influence of the compound of formula I (ie, negative control), 5 μL of distilled water is added to 4 wells.

All assays are incubated overnight (approximately 14-18 hours) at 37 ° C. 10 μL of Fast Blue B solution (0.5 mg / mL of Fast Blue B in buffer containing 0.1 M sodium acetate (pH 4.2) and 10% (v / v) Triton X-100) is added to each well. Quench the reaction, followed by shaking at room temperature for about 5 minutes. Plates can be analyzed with a Spectramax spectrophotometer or equivalent device (absorption maximum at 525 nm). By measuring DPP-IV activity over a compound concentration range of 10 nM to 3 μM, compound IC ₅₀ data can be obtained.

  The oral glucose tolerance test (“OGTT”) has been used in humans since at least the 1930s (Pincus et al., Am. J. Med. Sci, 188: 782 (1934)) in diagnosing human diabetes. Although it is used routinely, it is not used to evaluate the effects of therapeutic agents in patients.

  Glitazone (Fujita et al., Diabetes 32: 804810 (1983); Fujiwara et al., Diabetes 37: 1549-48 (1988); Izumi et al., Biopharm Dur. Dispos: 18: 247-257 (1997)) (Reddi et al., Diabet Metabl. 19: 44-51 (1993)), glucosidase inhibitors (Hamada et al., Jap. Pharmacol. Ther. 17: 17-28 (1988); Matsuo et al., Am. Clin. Nutr. 55: 314S-317S (1992)) and extrapancreatic effects of sulfonylureas (Kameda et al., Arzenim. Forsch./Drug Res. 32: 39044 (1982) KK mice have been used to evaluate Muller et al., Home.Metabl.Res.28: 469-487 (199)).

  KK mice are derived from an inbred line established for the first time by Kondo et al. (Kondo et al., Bull. Exp. Anim. 6: 107-112 (1957)). This mouse naturally develops an inherited form of polygenic diabetes that progresses and induces kidney, retinal and neurological complications similar to those seen in human diabetics, but the mouse Does not require insulin or other drugs for survival.

  The present invention is directed to the use of KK mice to evaluate the effects of insulin secretagogues in an oral glucose tolerance test.

In vivo analysis for glucose reduction The glucose-lowering effect of the DPP-IV inhibitor, the compound of formula I, can be demonstrated in 4-6 week old KK / H1J mice (Jackson Labs) in an oral glucose tolerance test. Mice are fasted overnight (approximately 14-18 hours), with free access to water. After fasting (time (“t” = 0), 25 μL of blood is collected from the retroorbital sinus and added to 0.025% heparinized saline (100 μL) on ice. 10 groups) are orally administered a solution of the compound of formula I (0.2 mL / mouse) in 0.5% methylcellulose, and the two control groups receive only 0.5% methylcellulose. At 15 minutes, blood is drawn from the mice as described above, followed by administration of 1 mg / kg of glucose in distilled water (0.2 mL / mouse) and the first control group is administered with glucose. Water is administered to control group 2. Blood is drawn again from mice as described above at time t = 45 minutes, blood samples are centrifuged, plasma collected, and glucose-containing on a Roche-Hitachi 912 glucose analyzer. Minutes The data can be expressed as percent inhibition of glucose excursion relative to the two control groups (ie, glucose levels in animals that were administered glucose but not the test compound) , Showing 0% inhibition, and glucose concentration in animals receiving water alone shows 100% inhibition).

General Experimental Procedure Melting points are determined with a Thomas Scientific capillary melting point apparatus and are not corrected.

  Flash chromatography was performed using W.W. C. This was performed by the method described by Still et al. (J. Org. Chem. 1978, 43, 2923).

The following examples are intended to detail specific embodiments of the invention and are not intended to limit the specification, including the claims in any way. The compounds exemplified in Examples 1 to 16 below showed in vitro activity showing an IC ₅₀ (test compound concentration required for 50% inhibition) of 3 μM or less.

(2S, 3S) -2-Amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyridin-1-yl) -pentan-1-one Step 1: [(1S, 2S)- 2-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidine-1-carbonyl) -butyl] -carbamic acid t-butyl ester (L) -Boc-isoleucine (322 mg, 1.30 mmol), Dichloromethane (10 mL) consisting of 3,3,4,4-tetrafluoro-pyrrolidine hydrochloride (300 mg, 1.67 mmol), hydroxybenzotriazole (225 mg, 1.67 mmol) and triethylamine (0.23 mL, 1.67 mmol) ) Was added to 1-(-3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (319 mg, 1.67 mL). Mol) was added. The mixture was stirred at room temperature overnight, diluted with ethyl acetate, washed with 2N HCl, saturated sodium bicarbonate solution, water and brine, dried over magnesium sulfate and concentrated. The product was purified by flash chromatography (hex / ethyl acetate, 4: 1) and isolated as a white solid (415 mg, 86%).

  The 3,3,4,4-tetrafluoro-pyrrolidine hydrochloride utilized in Step 1 was prepared according to Chaudry et al. Chem. Soc. 1964, p. 874. Alternatively, the fluorinated pyrrolidine can be prepared as described in Scheme II.

Step 2: Hydrochloric acid (2S, 3S) -2-amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyridin-1-yl) -pentan-1-one [(1S, 2S) 2-Methyl-1- (3,3,4,4-tetrafluoro-pyrrolidine-1-carbonyl) -butyl] -carbamic acid t-butyl ester (200 mg, 0.56 mmol) in ethyl acetate (4 mL) Dissolved, cooled to 0 ° C. and treated with gaseous HCl for about 1 minute. After 15 minutes at 0 ° C. and 30 minutes at room temperature, the mixture was concentrated to dryness and the solid was triturated with hexane, collected and dried under vacuum overnight (124 mg, 76%, melting point> 250 ° C.).

(2S, 3S) -2-Amino-1- (3-fluoro-azetidin-1-yl) -3-methyl-pentan-1-one Step 1: [(1S, 2S) -1- (3-hydroxy- Azetidine-1-carbonyl) -2-methyl-butyl] -carbamic acid t-butyl ester As described in Step 1 of Example 1, from a mixture of (L) -Boc-isoleucine and 3-hydroxyazetidine, [(1S, 2S) -1- (3-Hydroxy-azetidine-1-carbonyl) -2-methyl-butyl] -carbamic acid t-butyl ester was prepared. 3-Hydroxyazetidine is described in Lee, J. et al. Et al., (Bioorg. Med. Chem. Left. 2000, 10, 1063).

Step 2: [(1S, 2S) -1- (3-Fluoro-azetidine-1-carbonyl) -2-methyl-butyl] -carbamic acid t-butyl ester diethylaminosulfur trifluoride (0.46 mL, 3.5 Mmol) in ice-cold (−78 ° C.) dichloromethane (6 mL), [(1S, 2S) -1- (3-hydroxy-azetidine-1-carbonyl) -2-methyl-butyl] -t-butyl carbamate. A solution of the ester (1.0 g, 3.5 mmol) in dichloromethane (4 mL) was added dropwise. The mixture was warmed to room temperature and stirred overnight, then diluted with ethyl acetate and poured into ice / water. The layers were separated and the organic phase was washed with water and brine, dried over magnesium sulfate and concentrated. The product was purified by flash chromatography (ethyl acetate) and isolated as an oil (250 mg, 25%).

Step 3: (2S, 3S) -2-Amino-1- (3-fluoro-azetidin-1-yl) -3-methyl-pentan-1-one As described in Step 2 of Example 1, [( 1S, 2S) -1- (3-Fluoro-azetidine-1-carbonyl) -2-methyl-butyl] -carbamic acid t-butyl ester was treated with HCl to give (2S, 3S) -2-amino-1 -(3-Fluoro-azetidin-1-yl) -3-methyl-pentan-1-one was prepared (mp. 208-210 ° C).

(S) -2-Amino-2-cyclohexyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -ethanone Step 1: (S)-[1-Cyclohexyl-2-oxo- 2- (3,3,4,4-Tetrafluoro-pyrrolidin-1-yl) -ethyl] -carbamic acid t-butyl ester As described in Step 1 of Example 1, (L) -Boc-cyclohexylglycine And 3,3,4,4-tetrafluoropyrrolidine to (S)-[1-cyclohexyl-2-oxo-2- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -ethyl] -Carbamate t-butyl ester was prepared.

Step 2: (S) -2-Amino-2-cyclohexyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -ethanone As described in Step 2 of Example 1, ( S)-[1-cyclohexyl-2-oxo-2- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -ethyl] -carbamic acid t-butyl ester is treated with HCl to give ( S) -2-amino-2-cyclohexyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -ethanone was obtained (melting point 278 ° C.).

(2S, 3R) -2-Amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -pentan-1-one Step 1: [(1S, 2R)- 2-Methyl-1- (3,3,4,4-tetrafluoro-pyrrolidine-1-carbonyl) -butyl] -carbamic acid t-butyl ester As described in Step 1 of Example 1, (L)- From Boc-allo-isoleucine and 3,3,4,4-tetrafluoropyrrolidine, [(1S, 2R) -2-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidine-1-carbonyl) -Butyl] -carbamic acid t-butyl ester was prepared.

Step 2: (2S, 3R) -2-Amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -pentan-1-one of Step 2 of Example 1 Treat [(1S, 2R) -2-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidine-1-carbonyl) -butyl] -carbamic acid t-butyl ester as described. (2S, 3R) -2-amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -pentan-1-one (melting point> 250) ° C).

(S) -2-Amino-2-cyclohexyl-1- (3-fluoro-azetidin-1-yl) -ethanone As described in Example 2, from 3-azetidinol hydrochloride and (L) -Boc-cyclohexylglycine. (S) -2-amino-2-cyclohexyl-1- (3-fluoro-azetidin-1-yl) -ethanone was synthesized (melting point: 238-240 ° C.).

(S) -2-Amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -butan-1-one As described in Example 1, (L) From -Boc-valine and 3,3,4,4-tetrafluoropyrrolidine to (S) -2-amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -Butan-1-one was prepared (mp 234-238 ° C).

(2S, 3R) -2-Amino-1- (3,3-difluoro-pyrrolidin-1-yl) -3-methyl-pentan-1-one As described in Example 1, 3,3-difluorohydrochloride Pyrrolidine (prepared according to Giardina, G. Synlett 1995, page 55) and (L) -Boc-isoleucine from (2S, 3R) -2-amino-1- (3,3-difluoro-pyrrolidin-1-yl) -3-Methyl-pentan-1-one was prepared (mp 196-198 ° C).

(2S, 3S) -2-Amino-1- (3,3-difluoro-pyrrolidin-1-yl) -3-methyl-pentan-1-one As described in Example 7, 3,3-difluorohydrochloride (2S, 3S) -2-amino-1- (3,3-difluoro-pyrrolidin-1-yl) -3-methyl-pentan-1-one was prepared from pyrrolidine and (L) -Boc-isoleucine ( Mp 195-198 ° C).

Hydrochloric acid (S) -2-amino-4-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -pentan-1-one As described in Example 1, (L ) -Boc-leucine and 3,3,4,4-tetrafluoropyrrolidine hydrochloride to (S) -2-amino-4-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidine-1 hydrochloride -Yl) -Pentan-1-one was prepared (melting point> 250 ° C).

Hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (3,3-difluoro-azetidin-1-yl) -ethanone 3,3-difluoro-azetidine hydrochloride (International Publication No. 0047582 pamphlet) and Boc- ( L) -cyclohexylglycine followed by HCl deprotection as described in Example 1 to give hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (3,3-difluoro -Azetidin-1-yl) -ethanone was prepared (melting point 220 ° C. decomposition)

Hydrochloric acid (S) -2-amino-1- (3,3-difluoro-azetidin-1-yl) -3-methyl-pentan-1-one 3,3-difluoro-azetidine hydrochloride (International Publication No. 0047582 pamphlet) And Boc- (L) -isoleucine followed by HCl deprotection as described in Example 1 to give hydrochloric acid (S) -2-amino-1- (3,3-difluoro- Azetidin-1-yl) -3-methyl-pentan-1-one was prepared (melting point 195 ° C. decomposition).

Hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (4,4-difluoro-piperidin-1-yl) -ethanone 4,4-difluoro-piperidine-1-carboxylic acid t-butyl ester (International Publication No. Hydrochloric acid 4,4-difluoro-piperidine obtained by treating HCl (described in pamphlet No. 0040561) with Hoc was coupled with Boc- (L) -cyclohexylglycine as described in Example 1, followed by HCl desorption. Protected (melting point> 250 ° C.).

Hydrochloric acid (S) -2-amino-1- (4,4-difluoro-piperidin-1-yl) -3-methyl-pentan-1-one As described in Example 1, hydrochloric acid 4,4-difluoro- Hydrochloric acid (S) -2-amino-1- (4,4-difluoro-piperidin-1-yl) -3-methyl-pentan-1-one was prepared from piperidine and Boc- (L) -isoleucine (melting point) 190-192 ° C).

Hydrochloric acid (S) -2-amino-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -propan-1-one As described in Example 1, Boc- (L)- From alanine and 3,3,4,4-tetrafluoropyrrolidine, hydrochloric acid (S) -2-amino-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -propan-1-one Was prepared (melting point> 250 ° C.).

Another preparation of 3,3,4,4-tetrafluoropyrrolidine hydrochloride Step 1: Trifluoromethanesulfonic acid 2,2,3,3-tetrafluoro-4- (trifluoromethanesulfonyloxy) -butyl ester 2,2,3 Trifluoromethanesulfonic anhydride (34 mL, 200 mmol) was added dropwise to an ice-cold (0 ° C.) dichloromethane (250 mL) solution of 1,3-tetrafluorobutanediol (15 g, 93 mmol) and pyridine (19 mL, 230 mmol). did. After the addition, the mixture was stirred at 0 ° C. for 1 hour, followed by another 1 hour at room temperature, then diluted with dichloromethane, washed with water and brine, dried over magnesium sulfate, filtered, Concentration to near dry left a dichloromethane-containing oil.

Step 2: 1-Benzyl-3,3,4,4-tetrafluoro-pyrrolidine hydrochloride Crude trifluoro-methanesulfonic acid 2,2,3,3-tetrafluoro-4- (trifluoro-methanesulfonyloxy) -butyl An ethanol (230 mL) solution consisting of ester, benzylamine (10 mL, 93 mmol) and triethylamine (33 mL, 230 mmol) was heated to reflux overnight. The mixture was concentrated to about 1/3 volume, diluted with ether, washed with 1N sodium hydroxide, water and brine, dried over magnesium sulfate, filtered and concentrated to an oil. This oil was redissolved in ether, cooled to 0 ° C. and saturated with hydrogen chloride. The precipitate was collected and dried (23.8 g, 95%, mp 139-143 ° C.).

Step 3: 3,3,4,4-Tetrafluoro-pyrrolidine hydrochloride 1-benzyl-3,3,4,4-tetrafluoro-pyrrolidine hydrochloride containing 10% palladium on carbon (23.8 g , 88 mmol) in ethanol (300 mL) was treated with hydrogen at 45 psi in a Parr shaker for 3 hours. The mixture was filtered through Celite® and the filtrate was concentrated to dryness. The residue was triturated with ether and the solid collected and dried (13.4 g, 85%, mp 193-196 ° C.).

Claims (15)

A compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug:

[In the above formula,
R ¹ is 3-fluoroazetidin-1-yl, 3,3-difluoroazetidin-1-yl, 3,4-difluoropyrrolidin-1-yl, 3,3,4-trifluoropyrrolidin-1-yl 3,3,4,4-tetrafluoropyrrolidin-1-yl, 3-fluoropiperidin-1-yl, 4-fluoropiperidin-1-yl, 3,4-difluoropiperidin-1-yl, 3,5- Difluoropiperidin-1-yl, 3,3-difluoropiperidin-1-yl, 4,4-difluoropiperidin-1-yl, 3,4,5-trifluoropiperidin-1-yl, 3,3,4-tri Fluoropiperidin-1-yl, 3,3,5-trifluoropiperidin-1-yl, 3,4,4-trifluoropiperidin-1-yl, 3,3,4,5-tetrafluoropipe Lysine-1-yl, 3,4,4,5-tetrafluoropiperidin-1-yl, 3,3,4,4-tetrafluoropiperidin-1-yl, 3,3,5,5-tetrafluoropiperidine- 1-yl, 3,3,4,5,5-pentafluoropiperidin-1-yl, 3,3,4,4,5-pentafluoropiperidin-1-yl or 3,3,4,4,5 5-hexafluoropiperidin-1-yl, and R ² is (C ₁ -C ₈ ) alkyl or (C ₃ -C ₈ ) cycloalkyl]. The compound according to claim 1, wherein R ¹ is 3-fluoroazetidin-1-yl or 3,3-difluoroazetidin-1-yl. R ¹ is 3,4-difluoropyrrolidin-1-yl, 3,3,4-trifluoropyrrolidin-1-yl, 3,3,4,4-tetrafluoropyrrolidin-1-yl. Compound described in 1. R ¹ is 3-fluoropiperidin-1-yl, 4-fluoropiperidin-1-yl, 3,4-difluoropiperidin-1-yl, 3,5-difluoropiperidin-1-yl, 3,3-difluoropiperidine 2. A compound according to claim 1 which is -1-yl or 4,4-difluoropiperidin-1-yl. R ¹ is 3,4,5-trifluoropiperidin-1-yl, 3,3,4-trifluoropiperidin-1-yl, 3,3,5-trifluoropiperidin-1-yl, 3,4, 4-trifluoropiperidin-1-yl, 3,3,4,5-tetrafluoropiperidin-1-yl, 3,4,4,5-tetrafluoropiperidin-1-yl, 3,3,4,4- Tetrafluoropiperidin-1-yl, 3,3,4,5-tetrafluoropiperidin-1-yl, 3,3,4,5,5-pentafluoropiperidin-1-yl, 3,3,4,4 The compound according to claim 1, which is 5-pentafluoropiperidin-1-yl or 3,3,4,4,5,5-hexafluoropiperidin-1-yl. Hydrochloric acid (2S, 3S) -2-amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -pentan-1-one,
Hydrochloric acid (2S, 3S) -2-amino-1- (3-fluoro-azetidin-1-yl) -3-methyl-pentan-1-one,
Hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -ethanone,
Hydrochloric acid (2S, 3R) -2-amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -pentan-1-one,
Hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (3-fluoro-azetidin-1-yl) -ethanone,
Hydrochloric acid (S) -2-amino-3-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -butan-1-one;
Hydrochloric acid (S) -2-amino-4-methyl-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -pentan-1-one;
Hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (3,3-difluoro-azetidin-1-yl) -ethanone,
Hydrochloric acid (2S, 3S) -2-amino-1- (3,3-difluoro-azetidin-1-yl) -3-methyl-pentan-1-one,
Hydrochloric acid (S) -2-amino-2-cyclohexyl-1- (4,4-difluoro-piperidin-1-yl) -ethanone,
Hydrochloric acid (2S, 3S) -2-amino-1- (4,4-difluoro-piperidin-1-yl) -3-methyl-pentan-1-one,
Hydrochloric acid (S) -2-amino-1- (3,3,4,4-tetrafluoro-pyrrolidin-1-yl) -propan-1-one,
Hydrochloric acid (S) -2-amino-1- (3,3-difluoro-azetidin-1-yl) -3-methyl-butan-1-one;
Hydrochloric acid (S) -2-amino-1- (3-fluoro-azetidin-1-yl) -3-methyl-butan-1-one,
Hydrochloric acid (2S, 3R) -2-amino-1- (3-fluoro-azetidin-1-yl) -3-methyl-pentan-1-one,
Hydrochloric acid (2S, 3R) -2-amino-1- (3,3-difluoro-azetidin-1-yl) -3-methyl-pentan-1-one,
Hydrochloric acid (S) -2-amino-2-cyclopentyl-1- (3,3-difluoro-azetidin-1-yl) -ethanone,
Hydrochloric acid (S) -2-amino-2-cyclopentyl-1- (3-fluoro-azetidin-1-yl) -ethanone or hydrochloric acid (S) -2-amino-2-cyclopentyl-1- (3,3,4 , 4-Tetrafluoro-pyrrolidin-1-yl) -ethanone. The compound of formula I is

2. The compound of claim 1 which is the S enantiomer shown as   A pharmaceutical composition comprising a therapeutically effective amount of a compound according to claim 1, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug and a pharmaceutically acceptable diluent or carrier. . A therapeutically effective amount of the first compound according to claim 1, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug and insulin or an insulin analog; insulinotropin; biguanide; α ₂ -antagonists or imidazolines; glitazones; aldose reductase inhibitors; glycogen phosphorylase inhibitors; sorbitol dehydrogenase inhibitors; fatty acid oxidation inhibitors; α-glucosidase inhibitors; β-agonists; phosphodiesterase inhibitors; Vanadate, vanadium complex or peroxovanadium complex; amylin antagonist; glucagon antagonist; growth hormone secretagogue; gluconeogenesis inhibitor; somatostatin analog; renal glucose inhibitor; A therapeutically effective amount of a second compound that is a lipolytic agent; comprising a prodrug of said second compound or a pharmaceutically acceptable salt of said second compound or prodrug of said second compound , Pharmaceutical composition.   10. The composition of claim 9, further comprising a pharmaceutically acceptable carrier or diluent.   A method of treatment for inhibiting dipeptidyl peptidase IV in a mammal, comprising a therapeutically effective amount of the compound of claim 1, its prodrug or the compound or a pharmaceutically acceptable salt of the prodrug. Administering to said mammal in need of inhibiting dipeptidyl peptidase IV.   A method of treating a condition mediated by inhibition of dipeptidyl peptidase IV in a mammal, comprising a therapeutically effective amount of the compound, prodrug thereof, or the pharmaceutical of the compound or the prodrug. Administering a pharmaceutically acceptable salt to said mammal suffering from a condition mediated by inhibition of dipeptidyl peptidase IV.   The condition to be treated is type 2 diabetes, metabolic syndrome, hyperglycemia, impaired glucose tolerance, grape diabetes, metabolic acidosis, cataract, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy Infertility due to type 1 diabetes, obesity, condition exacerbated by obesity, hypertension, hyperlipidemia, atherosclerosis, osteoporosis, osteopenia, fragility, bone loss, fracture, acute coronary syndrome, polycystic ovary syndrome Disease, progression of type 2 diabetes, chronic fatigue, epilepsy, bowel movement related diseases, ulcers, irritable bowel syndrome, inflammatory bowel syndrome, anxiety, depression, insomnia, chronic fatigue, epilepsy, eating disorders The method according to claim 12, wherein the pain is chronic pain or alcohol drinking.   14. The method of claim 13, wherein the condition being treated is type 2 diabetes. A method for identifying whether a drug is a DPP-IV inhibitor, comprising:
Administering the drug to a fasted diabetic KK / H1J mouse;
Loading KK / H1J mice with oral glucose;
Assessing the response of the mouse to a subsequent oral glucose load, wherein said agent is treated with type 2 diabetes, metabolic syndrome, hyperglycemia, impaired glucose tolerance, grape diabetes, metabolic acidosis, cataract, Diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, type 1 diabetes, obesity, condition worsened by obesity, hypertension, hyperlipidemia, atherosclerosis, osteoporosis, bone loss Disease, vulnerability, bone loss, fracture, acute coronary syndrome, infertility due to polycystic ovary syndrome, prevention of disease progression in type 2 diabetes, anxiety, depression, insomnia, chronic fatigue, epilepsy, eating disorders, chronic pain A method that can be identified as a therapeutic agent for alcohol drinking, bowel movement related diseases, ulcers, irritable bowel syndrome, inflammatory bowel syndrome or short bowel syndrome.