`(10) Patent N0.:
`Us 6,699,871 132
`a
`Edmondson et a1.
`45 Date of Patent:
`Mar. 2 2004
`
`
`USOO6699871B2
`
`(75)
`
`(54) BETA-AMINO HETEROCYCLIC
`DIPEPTIDYL PEPTIDASE INHIBITORS FOR
`THE TREATMENT 0R PREVENTION OF
`DIABETES
`Inventors: Scott D. Edmondson, New York, NJ
`(US); Michael H. Fisher, Ringoes, NJ
`(US); Dooseop Kim, Westfield, NJ
`(3:3, 3:11:21]? “3,2333? 5:33;:1:131:
`NI (13%) Ann F Weber, Scotch Plains,
`NZI (US)? Jin 01;- Xu Scétch Plains NJ
`(US)
`’
`y
`’
`’
`
`(73) Assignee: MCI‘Ck & Co., 1116-, Rahway, NJ (US)
`
`( * ) Notice:
`‘
`’
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 21 days.
`
`(21) Appl. No; 10/189,603
`.
`Ffled:
`
`(22)
`(65)
`
`Jul. 5! 2002
`Prior Publication Data
`US 2003/0100563 A1 IVIay 29, 2003
`
`(60)
`
`Related US. Application Data
`Provisional application No. 60/303,474, filed on Jul. 6,
`2001.
`
`(51)
`
`Int. Cl.7 .................. C07D 487/04; A61K 31/4985;
`A61P 3/04; A61P 3/10
`....................................... 514/249; 544/350
`(52) US. Cl.
`(58) Field of Search ........................... 544/350 514/249
`’
`4
`'
`References Cited
`
`(56)
`
`US PATENT DOCUMENTS
`5 939 560 A
`8/1999 Jenkins et al
`69166063 A
`12:,2000 Villhauer
`
`(5:303:51 B1
`10/2001 Demuth et a1,
`FOREIGN PATENT DOCUMENTS
`
`548/535
`54/423
`514/866
`
`W0
`W0
`$8
`W0
`W0
`
`WO 98/19998
`W0 00/34241
`$8 833233451 :1)
`WO 01/96295 A2
`W0 02/0560 A2
`
`5/1998
`6/2000
`13/3881
`12/2001
`1/2002
`
`OTHER PUBLICATIONS
`“Novel N—substituted—2—cyanopyrrolidines as potent inhibi—
`tors of dipeptidyl peptidase IV in the treatment of non—in-
`sulin—dependent diabetes mellitus”, Exp. Opin, Ther. Pat-
`ents, 10(12), pp. 193771942 (2000).
`Ashworth, et a1., “Z—Cyanopyrrolidides as Potent, Stable
`Inhibitors of Dipeptidyl Peptidase IV”, Bioorg. Med. Chem.
`Lett., V01. 6, No. 10, pp. 1163—1166, (1996).
`.
`.
`.
`Ashworth, et al., “4—Cyanoth1azolld1des as Very Potent,
`Stable Inhibitors of Dipeptidyl Peptidase IV”, Bioorg. Med.
`Chem. Lett., vol. 6, No. 22, pp. 2745—2748, (1996).
`
`Deacon, ct al., “Dipcptidyl peptidasc IV inhibition as an
`approach to the treatment and prevention of type 2 diabetes:
`a historical perspective", Biochem. Biophys. Res. Commun.
`VOL 294’ PP‘ 1‘4 (2002)
`.
`.
`Przmary ExammerWark I.. Berch
`Assistant Examiner—Kahsay Habte
`(74) Allurney,Agen/, urFirm—Philippc L~ Duretle; MCIVIH
`Winokur; J. Eric Thies
`
`(57)
`
`ABSTRACT
`
`The present invention is directed to compounds Which are
`inhibitors 0f the dil’elm'dy1 Peptidasc‘w enzyme (“DP-IV
`inhibitors”) and which are useful in the treatment or pre-
`vention of diseases in which the di e lid 1
`e tidase-IV
`P P
`y P P
`enzyme is involved, such as diabetes and particularly type 2
`diabetes. The invention is also directed to pharmaceutical
`compositions comprising these compounds and the use of
`these compounds and compositions in the prevention or
`treatment of such diseases in which the dipeptidylpfiptidaSE-
`IV enzyme is involved.
`
`W0
`
`WO 97/40832
`
`11/1997
`
`26 Claims, N0 Drawings
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`US 6,699,871 B2
`
`1
`BETA-AMINO HETEROCYCLIC
`DIPEPTIDYL PEPTIDASE INHIBITORS FOR
`THE TREATMENT 0R PREVENTION OF
`DIABETES
`
`CROSS REFERENCE TO RELATED
`APPLICATIONS
`
`This application claims priority under 35 U.S.C. §ll9(e)
`from Serial No. 60/303,474, filed Jul. 6, 2001.
`BACKGROUND OF TIIE INVENTION
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`correction of hyperglycemia. However, the two biguanides,
`phenformin and metformin, can induce lactic acidosis and
`nausea/diarrhea. Metformin has fewer side effects than
`phenformin and is often prescribed for the treatment of Type
`2 diabetes.
`
`The glitazones (i.e. 5 —benzylthiazolidine—2,4—diones) are a
`more recently described class of compounds with potential
`for ameliorating many symptoms of type 2 diabetes. These
`agents substantially increase insulin sensitivity in muscle,
`liver and adipose tissue in several animal models of type 2
`diabetes resulting in partial or complete correction of the
`elevated plasma levels of glucose without occurrence of
`hypoglycemia. The glitazones that are currently marketed
`are agonists of the peroxisome proliferator activated recep—
`tor (PPAR), primarily the PPAR-gamma subtype. PPAR-
`gamma agonism is generally believed to be responsible for
`the improved insulin sensititization that is observed with the
`glitazones. Newer PPAR agonists that are being tested for
`treatment of Type II diabetes are agonists of the alpha,
`gamma or delta subtype, or a combination of these, and in
`many cases are chemically different from the glitazones (i.e.,
`they are not thiazolidinediones). Serious side effects (e.g.
`liver toxicity) have occurred with some of the glitazones,
`such as troglitazone,
`Additional methods of treating the disease are still under
`investigation. New biochemical approaches that have been
`recently introduced or are still under development include
`treatment with alpha-glucosidase inhibitors (e.g. acarbose)
`and protein tyrosine phosphatase—1B (FTP—1B) inhibitors.
`Compounds that are inhibitors of the dipeptidyl
`peptidase-IV (“DP-IV” or “DPP-IV”) enzyme are also under
`investigation as drugs that may be useful in the treatment of
`diabetes, and particularly type 2 diabetes. See for example
`WO 97/40832, WO 98/19998, US. Pat. No. 5,939,560,
`Bioorg. Med. Chem. Lem, 6(10), 1163—1166 (1996); and
`Binnrg. filed. Chem. Lem, 6(22), 2745—2748 (1996). The
`usefulness of DP-IV inhibitors in the treatment of type 2
`diabetes is based on the fact that DP—IV in vivo readily
`inactivates glucagon like peptide-l (GLP-l) and gastric
`inhibitory peptide (GIP). GLP-1 and GIP are incretins and
`are produced when food is consumed. The incretins stimu—
`late production of insulin, Inhibition of DP-IV leads to
`decreased inactivation of the incretins, and this in turn
`results in increased effectiveness of the incretins in stimu—
`lating production of insulin by the pancreas, DP-IV inhibi-
`tion therefore results in an increased level of serum insulin.
`Advantageously, since the incretins are produced by the
`body only when food is consumed, DP-IV inhibition is not
`expected to increase the level of insulin at inappropriate
`times, such as between meals, which can lead to excessively
`low blood sugar (hypoglycemia). Inhibition of DP-IV is
`therefore expected to increase insulin without increasing the
`risk of hypoglycemia, which is a dangerous side effect
`associated with the use of insulin secretagogues.
`DP-IV inhibitors also have other therapeutic utilities, as
`discussed herein. DP—IV inhibitors have not been studied
`extensively to date, especially for utilities other than diabe-
`tes. New compounds are needed so that improved DP-IV
`inhibitors can be found for the treatment of diabetes and
`potentially other diseases and conditions.
`SUMMARY OF THE INVENTION
`
`The present invention is directed to compounds which are
`inhibitors of the dipeptidyl peptidase-IV enzyme (“DP-IV
`inhibitors”) and which are useful in the treatment or pre—
`vention of diseases in which the dipeptidyl peptidase-IV
`
`Diabetes refers to a disease process derived from multiple
`causative factors and characterized by elevated levels of
`plasma glucose or hyperglycemia in the fasting state or after
`administration of glucose during an oral glucose tolerance
`test. Persistent or uncontrolled hyperglycemia is associated
`with increased and premature morbidity and mortality. Often
`abnormal glucose homeostasis is associated both directly
`and indirectly with alterations of the lipid, lipoprotein and
`apolipoprotein metabolism and other metabolic and hemo-
`dynamic disease. Therefore patients with Type 2 diabetes
`mellitus are at especially increased risk of macrovascular
`and microvascular complications, including coronary heart
`disease, stroke, peripheral vascular disease, hypertension,
`nephropathy, neuropathy, and retinopathy. Therefore, thera-
`peutical control of glucose homeostasis, lipid metabolism
`and hypertension are critically important
`in the clinical
`management and treatment of diabetes mellitus.
`There are two generally recognized forms of diabetes. In
`type 1 diabetes, or
`insulin-dependent diabetes mellitus
`(IDDM), patients produce little or no insulin, the hormone
`which regulates glucose utilization. In type 2 diabetes, or
`noninsulin dependent diabetes mellitus (NIDDM), patients
`often have plasma insulin levels that are the same or even
`elevated compared to nondiabetic subjects; however, these
`patients have developed a resistance to the insulin stimulat-
`ing effect on glucose and lipid metabolism in the main
`insulin-sensitive tissues, which are muscle, liver and adipose
`tissues, and the plasma insulin levels, while elevated, are
`insufficient to overcome the pronounced insulin resistance.
`Insulin resistance is not primarily due to a diminished
`number of insulin receptors but to a post-insulin receptor
`binding defect that is not yet understood. This resistance to
`insulin responsiveness results in insufficient insulin activa-
`tion of glucose uptake, oxidation and storage in muscle and
`inadequate insulin repression of lipolysis in adipose tissue
`and of glucose production and secretion in the liver.
`The available treatments for type 2 diabetes, which have
`not changed substantially in many years, have recognized
`limitations. While physical exercise and reductions in
`dietary intake of calories will dramatically improve the
`diabetic condition, compliance with this treatment is very
`poor because of well—entrenched sedentary lifestyles and
`excess food consumption, especially of foods containing ,
`high amounts of saturated fat. Increasing the plasma level of
`insulin by administration of sulfonylureas (e.g. tolbutamide
`and glipizide) or meglitinide, which stimulate the pancreatic
`[3-cells to secrete more insulin, and/or by injection of insulin
`when sulfonylureas or meglitinide become ineffective, can
`result in insulin concentrations high enough to stimulate the
`very insulin-resistant
`tissues. However, dangerously low
`levels of plasma glucose can result from administration of
`insulin or
`insulin secretagogues (sulfonylure as or
`meglitinide), and an increased level of insulin resistance due
`to the even higher plasma insulin levels can occur. The
`biguanides increase insulin sensitivity resulting in some
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`US 6,699,871 B2
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`enzyme is involved, such as diabetes and particularly type 2
`diabetes. The invention is also directed to pharmaceutical
`compositions comprising these compounds and the use of
`these compounds and compositions in the prevention or
`tre atinent of such diseases in which the dipeptidyl peptidase-
`IV enzyme is involved.
`
`DETAILED DESCRIPTION OF THE
`IN VENT] ON
`
`The present invention is directed to compounds of the
`formula I:
`
`NH2
`
`0
`
`A’\/K/u\N/fiybk
`big/X,1
`
`wherein:
`
`Ar is phenyl which is unsubstituted or substituted with
`1—5 of R3, wherein R3 is independently selected from
`the group consisting of:
`(1) halogen,
`(2) Cmalkyl, which is linear or branched and is unsub—
`stituted or substituted with 1—5 halogens,
`(3) OC1_6alkyl, which is linear or branched and is
`unsubstituted or substituted with 175 halogens, and
`(4) CN;
`X is selected from the group consisting of:
`(1) N, and
`(2) CR2;
`R1 and R2 are independently selected from the group
`consisting of:
`(1) hydrogen,
`(2) CN.
`(3) C1_10all<yl, which is linear or branched and which is
`unsubstituted or substituted with 1—5 halogens or
`phenyl, which is unsubstituted or substituted with
`1—5 substituents independently selected from
`halogen, CN, OH, R“, 0R4, NHSOZR4, SOZR4,
`COZH, and COZCliéalkyl, wherein the COZClicalkyl
`is linear or branched,
`(4) phenyl which is unsubstituted or substituted with
`1—5 substituents independently selected from
`halogen, CN, OH, R4, 0R4, NHSOZR4, SOZR4,
`COZH, and C02C1_fialkyl, wherein the COszalkyl
`is linear or branched, and
`(6) a 5— or 6—membered heterocycle which may be
`saturated or unsaturated comprising 1—4 heteroatoms
`independently selected from N, S and 0, the hetero-
`cycle being unsubstituted or substituted with 173
`substituents independently selected from oxo, OH,
`halogen, CHjalkyl, and OCLSalkyl, wherein the
`C1_6alkyl and OC1_6alkyl are linear or branched and
`optionally substituted with 1—5 halogens;
`R4 is CLdalkyl, which is linear or branched and which is
`unsubstituted or substituted with 175 groups indepen—
`dently selected from halogen, COZII, and COZCL
`salkyl, wherein the COZCLfialkyl is linear or branched;
`and pharmaceutically acceptable salts thereof and indi—
`vidual diastereomers thereof.
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`An embodiment of the present invention includes com-
`pounds of the formula la:
`
`la
`
`wherein X, Ar and R1 are defined herein;
`and pharmaceutically acceptable salts and individual dias-
`tereomers thereof.
`Another embodiment of the present invention includes
`compounds of the formula Ib:
`
`Ib
`
`wherein Ar and R1 are defined herein;
`and pharmaceutically acceptable salts and individual dias-
`tereomers thereof.
`Another embodiment of the present invention includes
`compounds of the formula Ic:
`
`N112
`
`o
`
`ATMN/YN
`
`Ic
`
`wherein Ar, R1 and R2 are defined herein;
`and pharmaceutically acceptable salts thereof and iridi-
`vidual diastereomers thereof.
`In the present invention it is preferred that Ar is phenyl
`which is unsubstituted or substituted with 1—5 substitutents
`which are independently selected from the group consisting
`of:
`
`(l) fluoro,
`(2) bromo, and
`(3) CF3.
`In the present invention it is more preferred that AI is
`selected from the group consisting of:
`(1) phenyl,
`(2) 2-fluorophenyl,
`(3) 3,4—difluorophenyl,
`(4) 2,5-difluorophenyl,
`(5) 2,4,5-trifluorophenyl,
`(6) 2-fluoro-4-(triflouromethyl)phenyl, and
`(7) 4—bromo—2,5—difluorophenyl.
`In the present invention it is preferred that R1 is selected
`from the group consisting of:
`(1) hydrogen, and
`(2) C1_Gall<yl, which is linear or branched and which is
`unsubstituted or substituted with phenyl or 1—5 fluoro.
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`US 6,699,871 B2
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`5
`In the present invention it is more preferred that R1 is
`selected from the group consisting of:
`(1) hydrogen,
`(2) methyl,
`(3) ethyl,
`(4) CFg,
`(5) CHZCFB,
`(5) CFZCF3
`(6) phenyl, and
`(7) benzyl.
`In the present invention it is more preferred that R1 is
`selected from the group consisting of:
`(1) hydrogen,
`(2) methyl,
`(3) ethyl,
`(4) CF3, and
`(5) CHZCFB.
`In the present invention it is even more preferred that R1
`is hydrogen or CF3.
`In the present invention it is preferred that R2 is selected
`from:
`
`(1) hydrogen,
`(2) C1_6alkyl, which is linear or branched and which is
`unsubstituted or substituted with 1—5 fiuoro,
`(3) phenyl, which is unsubstituted or substituted with 1—3
`substituents independently selected from fiuoro, OCH3,
`and OCF3.
`In the present invention it is more preferred that R2 is
`selected from the group consisting of:
`(1) hydrogen,
`(2) methyl,
`(3) ethyl,
`(4) CF»
`(5) CH2CF3,
`(5) CFZCF3
`(6) phenyl,
`(7) (4-methoxy)phenyl,
`(8) (4-trifluorometlioxy)plienyl,
`(9) 4-fluorophenyl, and
`(10) 3,4-dilluorophenyl.
`In the present invention it is even more preferred that R2
`is CF3 or CFZFB.
`In the present invention it is preferred that R3 is F, Br or
`CFB.
`The compounds of the present invention may contain one
`or more asymmetric centers and can thus occur as racemates
`and racemic mixtures, single enantiomers, diastereomeric
`mixtures and individual diastereomers. The compounds of
`the instant invention have one asymmetric center at the beta
`carbon atom, Additional asymmetric centers may be present
`depending upon the nature of the various substituents on the
`molecule. Each such asymmetric center will independently
`produce two optical isomers and it is intended that all of the
`possible optical isomers and diastereomers in mixtures and
`as pure or partially purified compounds are included within
`the ambit of this invention, The present invention is meant
`to comprehend all such isomeric forms of these compounds.
`Some of the compounds described herein contain olefinic
`double bonds, and unless specified otherwise, are meant to
`include both E and Z geometric isomers.
`Some of the compounds described herein may exist as
`tautomers, which have different points of attachment of
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`hydrogen accompanied by one or more double bond shifts.
`For example, a ketone and its enol form are keto-enol
`tautomers. The individual tautomers as well as mixtures
`thereof are encompassed with compounds of the present
`invention.
`Formula I shows the structure of the class of compounds
`without preferred stereochemistry. Formula Ia shows the
`preferred sterochemistry at the carbon atom that is attached
`to the amine group of the beta amino acid from which these
`compounds are prepared.
`The independent syntheses of these diastereomers or their
`chromatographic separations may be achieved as known in
`the art by appropriate modification of the methodology
`disclosed herein. Their absolute stereochemistry may be
`determined by the x-ray crystallography of crystalline prod-
`ucts or crystalline intermediates which are derivatized, if
`necessary, with a reagent containing an asymmetric center of
`known absolute configuration.
`If desired, racemic mixtures of the compounds may be
`separated so that the individual enantiomers are isolated.
`The separation can be carried out by methods well known in
`the art, such as the coupling of a racemic mixture of
`compounds to an enantiomerically pure compound to form
`a diastereomeric mixture, followed by separation of the
`individual diastereomers by standard methods, such as frac—
`tional crystallization or chromatography. The coupling reac-
`tion is often the formation of salts using an enantiomerically
`pure acid or base. The diasteromeric derivatives may then be
`converted to the pure enantiomers by cleavage of the added
`chiral residue. The racemic mixture of the compounds can
`also be separated directly by chromatographic methods
`utilizing chiral stationary phases, which methods are well
`known in the art.
`Altematively, any enantiomer of a compound may be
`obtained by stereoselcctive synthesis rising optically pure
`starting materials or reagents of knoan configuration by
`methods well known in the art.
`The term “pharmaceutically acceptable salts” refers to
`salts prepared from pharmaceutically acceptable non-toxic
`bases or acids including inorganic or organic bases and
`inorganic or organic acids. Salts derived from inorganic
`bases include aluminum, ammonium, calcium, copper,
`ferric,
`ferrous,
`lithium, magnesium, manganic salts,
`manganous, potassium, sodium, zinc, and the like. Particu-
`larly preferred are the ammonium, calcium, magnesium,
`potassium, and sodium salts. Salts in the solid form may
`exist in more than one crystal structure, and may also be in
`the form of hydrates. Salts derived from pharmaceutically
`acceptable organic non-toxic bases include salts of primary,
`secondary, and tertiary amines, substituted amines including
`naturally occurring substituted amines, cyclic amines, and
`basic ion exchange resins, such as arginine, betaine, caffeine,
`choline, N,N’-dibenzylethylene-diamine, diethylamine,
`2—diethy1aminoethanol, 2—dimethylaminoethanol,
`ethanolamine, ethylcncdiamine, N-cthyl-morpholinc,
`N-ethylpiperidine, glucamine, glucosamine, histidine,
`hydrabamine,
`isopropylamine,
`lysine, methylglucamine,
`morpholine, piperazine, piperidine, polyamine resins,
`procaine, purines,
`tlieobromine,
`triethylamine,
`trimethylamine, tripropylamine, tromethamine, and the like.
`When the compound of the present invention is basic,
`salts may be prepared from pharmaceutically acceptable
`non—toxic acids, including inorganic and organic acids. Such
`acids include acetic, benzenesulfonic, benzoic,
`camphorsulfonic, citric, ethanesulfonic, [umaric, gluconic,
`glutamic, hydrobromic, hydrochloric,
`isethionic,
`lactic,
`maleie, malic, mandelic, methanesulfonic, mucie, nitric,
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`US 6,699,871 B2
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`7
`pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric,
`p-toluenesulfonic acid, and the like. Particularly preferred
`are citric, hydrobromic, hydrochloric, maleic, phosphoric,
`sulfuric, fumaric, and tartaric acids.
`It will be understood that, as used herein, references to the
`compounds of Formula I are meant
`to also include the
`pharmaceutically acceptable salts.
`As appreciated by those of skill in the art, halo or halogen
`as used herein are intended to include fiuoro, chloro, bromo
`and iodo. Similarly, CHg, as in Cl_8alkyl
`is defined to
`identify the group as having 1, 2, 3, 4, 5, 6, 7 or 8 carbons
`in a linear or branched arrangement, such that C1 _Ralkyl
`specifically includes methyl, ethyl, n-propyl,
`iso-propyl,
`n—butyl, iso—butyl, tert—butyl, pentyl, hexyl, heptyl and octyl.
`Likewise, Co, as in Coalkyl is defined to identify the pres-
`ence of a direct covalent bond. A group which is designated
`as being independently substituted with substituents may be
`independently substituted with multiple numbers of such
`substituents. The term “heterocycle” as used herein is
`intended to include 5— or 6—membered ring systems which
`are within the following listing: benzimidazolyl,
`be nzodioxanyl, benzofuranyl, benzopyrazolyl,
`benzothiadiazolyl, benzotriazolyl, benzothiophenyl,
`benzoxadiazolyl, benzoxazolyl, carbazolyl, carbolinyl,
`chromanyl, cinnolinyl,
`furanyl,
`imidazolyl,
`indolinyl,
`indolyl, indolazinyl, indazolyl, isohenzofuranyl, isoindolyl,
`isoquinolyl,
`isothiazolyl,
`isoxazolyl, naphthyridinyl,
`oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridopyridinyl,
`pyridazinyl, pyridyl, pyrimidyl, pyrrolyl, quinazolinyl,
`quino linyl, quinoxalinyl,
`tetrazo lyl,
`thiadiazolyl,
`thiazolidinyl,
`thiazolyl,
`thienyl,
`triazolyl, azetidinyl, 1,4—
`dioxanyl, hexahydroazepinyl, piperazinyl, piperidinyl,
`pyrrolidinyl,
`rnorpholinyl,
`thiornorpholinyl,
`dihydrobenzimidazolyl, dihydrobenzofuranyl,
`dihydrobenzothiophenyl, dihydrobc nzoxazolyl,
`dihydrofuranyl, dihydroimidazolyl, dihydroindolyl,
`dihydroisooxazolyl,
`dihydroisothiazolyl,
`dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl,
`dihydropyrazolyl, dihydropyridinyl, dihydropyrirnidinyl,
`dihydropyrrolyl, dihydroquinolinyl, dihydrotetrazolyl,
`dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl,
`dihydrotriazolyl,
`dihydroazetidinyl,
`methylenedioxybenzoyl,
`tetrahydrofuranyl ,
`tetrahydroimidazolyl, tetrahydroisoquinolinyl, and tetrahy-
`drothienyl.
`Exemplifying the invention is the use of the compounds
`disclosed in the Examples and herein.
`Specific compounds within the present invention include
`a compound which selected from the group consisting of the
`compounds disclosed in the following Examples and phar-
`maceutically acceptable salts thereof and individual diaste—
`reomers thereof.
`The subject compounds are useful in a method of inhib-
`iting the dipeptidyl peptidase—IV enzyme in a patient such as
`a mammal in need of such inhibition comprising the admin-
`istration of an effective amount of the compound. The
`present invention is directed to the use of the compounds
`disclosed herein as inhibitors of dipeptidyl peptidase-IV
`enzyme activity.
`In addition to primates, such as humans, a variety of other
`mammals can be treated according to the method of the
`present invention. For instance, mammals including, but not
`limited to, cows, sheep, goats, horses, dogs, cats, guinea
`pigs, rats or other bovine, ovine, equine, canine, feline,
`rodent or murine species can be treated. However,
`the
`method can also be practiced in other species, such as avian
`species (e.g., chickens).
`
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`8
`The present invention is further directed to a method for
`the manufacture of a medicament for inhibiting dipeptidyl
`peptidase-IV enzyme activity in humans and animals com-
`prising combining a compound of the present invention with
`a pharmaceutical carrier or diluent.
`The subject treated in the present methods is generally a
`mammal, preferably a human being, ma e or female,
`in
`
`whom inhibition of dipeptidyl peptidase—IV enzyme activity
`
`is desired. The term “therapeutically e eetive amount”
`means the amount of the subject compound that will elicit
`the biological or medical response of a tissue, system,
`animal or human that is being sought by the researcher,
`veterinarian, medical doctor or other clinician.
`The term “composition" as used herein is intended to
`encompass a product comprising the specified ingredients in
`the specified amounts, as well as any product which results,
`directly or indirectly, from combination of the specified
`ingredients in the specified amounts. Such term in relation to
`pharmaceutical composition,
`is intended to encompass a
`product comprising the active ingredient(s), and the inert
`ingredient(s) that make up the carrier, as well as any product
`which results, directly or indirectly, from combination, com-
`plexation or aggregation of any two or more of the
`ingredients, or from dissociation of one or more of the
`ingredients, or from other types of reactions or interactions
`of one or more of the ingredients. Accordingly, the pharma-
`ceutical compositions of the present invention encompass
`any composition made by admixing a compound of the
`present invention and a pharmaceutically acceptable carrier.
`By “pharmaceutically acceptable” it is meant the carrier,
`diluent or excipient must be compatible with the other
`ingredients of the formulation and not deleterious to the
`recipient thereof.
`The terms “administration of” and or “administering a”
`compound should he understood to mean providing a com-
`pound of the invention or a prodrug of a compound of the
`invention to the individual in need of treatment.
`The utility of the compounds in accordance with the
`present invention as inhibitors of dipeptidyl peptidase-IV
`enzyme activity may be demonstrated by methodology
`known in the art. Inhibition constants are determined as
`follows. A continuous fiuorornetric assay is employed with
`the substrate Gly—Pro—AMC, which is cleaved by DP—IV to
`release the fluorescent AMC leaving group. The kinetic
`parameters that describe this reaction are as follows: Km=50
`‘uM; kcm=75 s'l; kmt/Km=1.5x106 M'ls'l. A typical reac-
`tion contains approximately 50 pM enzyme, 50 ,uM Gly-
`Pro—AMC, and bulfer (100 mM HEPES, pH 7.5, 0.1 mg/ml
`BSA) in a total reaction volume of 100 ill. liberation of
`AMC is monitored continuously in a 96-well plate fluorom-
`eter using an excitation wavelength of 360 nm and an
`emission wavelength of 460 nm. Under these conditions,
`approximately 0.8 ,uM AMC is produced in 30 minutes at 25
`degrees C. The enzyme used in these studies was soluble
`(transmemhrane domain and cytoplasmic extension
`excluded) human protein produced in a baculovirus expres-
`sion system (Bac—To—Bac, Gibco BRL). The kinetic con—
`stants for hydrolysis of Gly-Pro-AMC and GLP-1 were
`found to be in accord with literature values for the native
`enzyme. To measure the dissociation constants for
`compounds, solutions of inhibitor in DMSO were added to
`reactions containing enzyme and substrate (final DMSO
`concentration is 1%). All experiments were conducted at
`room temperature using the standard reaction conditions
`described above. To determine the dissociation constants
`(K,), reaction rates were fit by non—linear regression to the
`Michaelis-Menton equation for competitive inhibition. The
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`9
`errors in reproducing the dissociation constants are typically
`less than two-fold.
`In particular, the compounds of the following examples
`had activity in inhibiting the dipeptidyl peptidase-IV
`enzyme in the aforementioned assays, generally with an
`1C5O of less than about 1 ‘uM. Such a result is indicative of
`the intrinsic activity of the compounds in use as inhibitors
`the dipeptidyl peptidase—IV enzyme activity.
`Dipeptidyl peptidase-IV enzyme (DP-IV) is a cell surface
`protein that has been implicated in a wide range of biological
`functions. It has a broad tissue distribution (intestine, kidney,
`liver, pancreas, placenta, thymus, spleen, epithelial cells,
`vascular endothelium, lymphoid and myeloid cells, serum),
`and distinct tissue and cell—type expression levels. DP—IV is
`identical to the T cell activation marker CD26, and it can
`cleave a number of immunoregulatory, endocrine, and neu-
`rological peptides in vitro. This has suggested a potential
`role for this peptidase in a variety of disease processes in
`humans or other species.
`in a
`Accordingly,
`the subject compounds are useful
`method for the prevention or treatment of the following
`diseases, disorders and conditions.
`Type II Diabetes and Related Disorders: It is well estab—
`lished that the incretins GLP-1 and GIP are rapidly inacti-
`vated in vivo by DP—IV. Studies with DP—IV('/')—deficient
`mice and preliminary clinical
`trials indicate that DP-IV
`inhibition increases the steady state concentrations of GLP-1
`and GIP, resulting in improved glucose tolerance. By anal—
`ogy to GLP-1 and GIP, it is likely that other glucagon family
`peptides involved in glucose regulation are also inactivated
`by DP—IV (eg. PACAP, glucagon). Inactivation of these
`peptides by DP-IV may also play a role in glucose homeo-
`stasis.
`The DP—IV inhibitors of the present invention therefore
`have utility in the treatment of type II diabetes and in the
`treatment and prevention of the numerous conditions that
`often accompany Type II diabetes,
`including metabolic
`syndrome X, reactive hypoglycemia, and diabetic dyslipi-
`demia. Obesity, discussed below, is another condition that is
`often found with Type II diabetes that may respond to
`treatment with the compounds of this invention.
`The following diseases, disorders and conditions are
`related to Type 2 diabetes, and therefore may be treated,
`controlled or in some cases prevented, by treatment with the
`compounds of this invention: (1) hyperglycemia, (2) low
`glucose tolerance, (3) insulin resistance, (4) obesity, (5) lipid
`disorders,
`(6) dyslipidemia,
`(7) hyperlipidemia,
`(8)
`hypertriglyceridemia, (9) hypercholesterolemia, (10) low
`HDI. levels, (11) high IDI. levels, (1 2) atherosclerosis and
`its sequelae, (13) vascular restenosis, (14) irritable bowel
`syndrome,
`(15)
`inflammatory bowel disease,
`including
`Crohn’s disease and ulcerative colitis, (16) other inflamma-
`tory conditions, (17) pancreatitis, (18) abdominal obesity,
`(19) neurodegenerative disease,
`(20)
`retinopathy,
`(21)
`nephropathy, (22) neuropathy, (23) Syndrome X, (24) ova-
`rian hyperandrogenism (polycystic ovarian syndrome), and
`other disorders where insulin resistance is a component.
`Obesity: DP-IV inhibitors may be useful for the treatment of
`obesity. This is based on the observed inhibitory effects on
`food intake and gastric emptyng of GLP—1 and GLP—2.
`Exogenous administration of GLP-1 in humans significantly
`decreases food intake and slows gastric emptying (Am. J.
`Physiol. 277, R9104R916 (1999)). ICV administration of
`GLP-1 in rats and mice also has profound effects on food
`intake (Nature Medicine 2, 1254—1258 (1996)). This inhi-
`bition of feeding is not observed in GLP—1RH’) mice,
`indicating that these effects are mediated through brain
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`it is likely that
`GLP-l receptors. By analogy to GLP-l,
`GLP-2 is also regulated by DP-IV. ICV administration of
`GIP-2 also inhibits food intake, analogous to the effects
`observed with GLP-1 (Nature Medicine 6, 802—807 (2000)).
`Growth Hormone Deficiency: DP—IV inhibition may be
`useful for the treatment of growth hormone deficiency, based
`on the hypothesis that gowth-hormone releasing factor
`(GRF), a peptide that stimulates release of growth hormone
`from the anterior pituitary, is cleaved by the DP-IV enzyme
`in vivo (WO 0066297). The following data provide evi-
`dence that GRF is an endogenous substrate: (1) GRF is
`efficiently cleaved in vitro to generate the inactive product
`GRb'[3-44] (BBA 1122, 147—153 (1992)); (2) GRF is rapidly
`degraded in plasma to GRF[3—44]; this is prevented by the
`DP-IV inhibitor diprotin A; and (3) GRF[3-44] is found in
`the plasma of a human GRF transgenic pig (J. Clin. Invest.
`83, 153341540 (1989)). Thus DP—IV inhibitors may be
`useful for the same spectrum of indications which have been
`considered for growth hormone secretagogues.
`Intestinal Injury: The potential for using DP—IV inhibitors
`for the treatment of intestinal injury is suggested by the
`results of studies indicating that glucagon-like peptide-2
`(GLP—2), a likely endogenous substrate for DP—IV, may
`exhibit
`trophic effects on the intestinal epithelium
`(Regulatory Peptides 90, 27—32 (2000)). Administration of
`GI .P-2 results in increased small bowel mass in rodents and
`attenuates intestinal injury in rodent models of colitis and
`enteritis.
`Immunosuppression: DP-IV inhibition may be useful for
`